WO2008024725A1 - Aza-benzofuranyl compounds and methods of use - Google Patents

Aza-benzofuranyl compounds and methods of use Download PDF

Info

Publication number
WO2008024725A1
WO2008024725A1 PCT/US2007/076344 US2007076344W WO2008024725A1 WO 2008024725 A1 WO2008024725 A1 WO 2008024725A1 US 2007076344 W US2007076344 W US 2007076344W WO 2008024725 A1 WO2008024725 A1 WO 2008024725A1
Authority
WO
WIPO (PCT)
Prior art keywords
alkyl
mmol
nhc
formula
compound
Prior art date
Application number
PCT/US2007/076344
Other languages
French (fr)
Inventor
Stephen Price
Karen Williams
Pascal Pierre Savy
Hazel Joan Dyke
John Gary Montana
Mark S. Stanley
Liang Bao
Original Assignee
Genentech, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority to AT07814272T priority Critical patent/ATE531720T1/en
Application filed by Genentech, Inc. filed Critical Genentech, Inc.
Priority to ES07814272T priority patent/ES2376771T3/en
Priority to EP07814272A priority patent/EP2069354B1/en
Priority to CA002660546A priority patent/CA2660546A1/en
Priority to BRPI0714635-3A priority patent/BRPI0714635A2/en
Priority to JP2009525720A priority patent/JP5448818B2/en
Priority to AU2007286808A priority patent/AU2007286808B2/en
Priority to MX2009001878A priority patent/MX2009001878A/en
Priority to CN200780039152.0A priority patent/CN101605794B/en
Priority to KR1020097005786A priority patent/KR101428116B1/en
Publication of WO2008024725A1 publication Critical patent/WO2008024725A1/en
Priority to IL196956A priority patent/IL196956A/en
Priority to NO20091197A priority patent/NO20091197L/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/04—Ortho-condensed systems
    • C07D491/044—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring
    • C07D491/048—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring the oxygen-containing ring being five-membered
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/4355—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having oxygen as a ring hetero atom
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/50—Pyridazines; Hydrogenated pyridazines
    • A61K31/5025—Pyridazines; Hydrogenated pyridazines ortho- or peri-condensed with heterocyclic ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/18—Drugs for disorders of the alimentary tract or the digestive system for pancreatic disorders, e.g. pancreatic enzymes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/12—Drugs for disorders of the urinary system of the kidneys
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

  • the invention relates to azabenzofuranyl compounds with anti-cancer and/or anti-inflammatory activity and more specifically to azabenzofuranyl compounds which inhibit MEK kinase activity.
  • the invention also relates to methods of using the compounds for in vitro, in situ, and in vivo diagnosis or treatment of mammalian cells, or associated pathological conditions.
  • MAP (mitogen-activated protein) kinase (MAPK) pathway has emerged as the crucial route between membrane-bound Ras and the nucleus.
  • the MAPK pathway encompasses a cascade of phosphorylation events involving three key kinases, namely Raf, MEK (MAP kinase kinase) and ERK (MAP kinase).
  • Active GTP -bound Ras results in the activation and indirect phosphorylation of Raf kinase.
  • Raf then phosphorylates MEKl and 2 on two serine residues (S218 and S222 for MEKl and S222 and S226 for MEK2) (Ahn et al., Methods in Enzymology 2001, 332, 417-431).
  • ERK phosphorylation by MEK occurs on Y204 and T202 for ERKl and Y 185 and T 183 for ERK2 (Ahn et al., Methods in Enzymology 2001, 332, 417-431). Phosphorylated ERK dimerizes and then translocates to the nucleus where it accumulates (Khokhlatchev et al., Cell 1998, 93, 605-615).
  • ERK is involved in several important cellular functions, including but not limited to nuclear transport, signal transduction, DNA repair, nucleosome assembly and translocation, and mRNA processing and translation (Ahn et al., Molecular Cell 2000, 6, 1343-1354). Overall, treatment of cells with growth factors leads to the activation of ERKl and 2 which results in proliferation and, in some cases, differentiation (Lewis et al., Adv. Cancer Res. 1998, 74, 49-139). [0007] There has been strong evidence that genetic mutations and/or overexpression of protein kinases involved in the MAP kinase pathway lead to uncontrolled cell proliferation and, eventually, tumor formation, in proliferative diseases.
  • some cancers contain mutations which result in the continuous activation of this pathway due to continuous production of growth factors.
  • Other mutations can lead to defects in the deactivation of the activated GTP -bound Ras complex, again resulting in activation of the MAP kinase pathway.
  • Mutated, oncogenic forms of Ras are found in 50% of colon and >90% pancreatic cancers as well as many others types of cancers (Kohl et al., Science 1993, 260, 1834-1837).
  • bRaf mutations have been identified in more than 60% of malignant melanoma (Davies, H. et al., Nature 2002, 417, 949-954).
  • MEK has emerged as an attractive therapeutic target in the MAP kinase cascade pathway.
  • MEK downstream of Ras and Raf, is highly specific for the phosphorylation of MAP kinase; in fact, the only known substrates for MEK phosphorylation are the MAP kinases, ERKl and 2.
  • MEK inhibitors have been shown to inhibit human tumor growth in nude mouse xenografts, (Sebolt-Leopold et al., Nature-Medicine 1999, 5 (7), 810-816); Trachet et al., AACR Apr. 6-10, 2002, Poster #5426; Tecle, H. IBC 2.sup.nd International Conference of Protein Kinases, Sep. 9-10, 2002), block static allodynia in animals (WO 01/05390 published Jan. 25, 2001) and inhibit growth of acute myeloid leukemia cells (Milella et al., J Clin Invest 2001, 108 (6), 851-859).
  • MEK inhibitors have also been discussed in, for example, WO02/06213, WO 03/077855 and WO03/077914.
  • MEK inhibitors As effective and safe therapeutics for treating a variety of proliferative disease states, such as conditions related to the hyperactivity of MEK, as well as diseases modulated by the MEK cascade.
  • the invention relates generally to aza-benzofuran compounds of Formula I
  • Certain hyperproliferative and inflammatory disorders are characterized by the modulation of MEK kinase function, for example by mutations or overexpression of the proteins. Accordingly, the compounds of the invention and compositions thereof are useful in the treatment of hyperproliferative disorders such as cancer and/or inflammatory diseases such as rheumatoid arthritis.
  • Z 2 is CR 2 or N
  • Z 3 is CR 3 or N
  • Z 4 is CR 4 or N; where one or two of Z 1 , Z 2 , Z 3 , and Z 4 are N;
  • R 5 and R 6 are independently selected from H or Ci-Ci 2 alkyl
  • X 2 is selected from carbocyclyl, heterocyclyl, aryl, and heteroaryl;
  • R 11 , R 12 and R 13 are independently H, Ci-Ci 2 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl, or R 11 and R 12 together with the nitrogen to which they are attached form a 3-8 membered saturated, unsaturated or aromatic ring having 0-2 heteroatoms selected from O, S and N, wherein said ring is optionally substituted with one or more groups selected from halo, CN, CF 3 , -OCF 3 , -NO 2 , Ci-C 6 alkyl, -OH, -SH, -0(Ci-C 6 alkyl), -S(Ci-C 6 alkyl), -NH 2 , -NH(Ci-C 6 alkyl), -N(Ci-C 6 alkyl) 2 , -SO 2 (Ci-C 6 alkyl), -CO 2 H, -CO
  • R 14 and R 15 are independently selected from H, Ci-Ci 2 alkyl, aryl, carbocyclyl, heterocyclyl, and heteroaryl; m and n are independently selected from O, 1, 2, 3, 4, 5, or 6;
  • R 19 and R 20 are independently selected from H, Ci-Ci 2 alkyl, -(CH 2 ) n -aryl, -(CH 2 ) n - carbocyclyl, -(CH 2 ) n -heterocyclyl, and -(CH 2 ) n -heteroaryl;
  • R 21 is C 1 -C 12 alkyl, C 2 -Cs alkenyl, C 2 -Cs alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl, wherein each member of R 21 is optionally substituted with one or more groups selected from halo, CN, -OCF 3 , CF 3 , -NO 2 , Ci-C 6 alkyl, -OH, -SH, -0(Ci-C 6 alkyl), -S(Ci-C 6 alkyl), -NH 2 , -NH(Ci-C 6 alkyl), -N(Ci-C 6 alkyl) 2 , -SO 2 (Ci-C 6 alkyl), -CO 2 H, -CO 2 (Ci-C 6 alkyl), -C(O)NH 2 , -C(0)NH(C r C 6 alkyl), -C(0)N(C r C 6 alkyl) 2 ,
  • R 22 is H or Ci-Ci 2 alkyl.
  • the present invention includes a composition (e.g., a pharmaceutical composition) comprising a compound of Formula I (and/or solvates, hydrates and/or salts thereof) and a carrier (a pharmaceutically acceptable carrier).
  • a composition e.g., a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of Formula I (and/or solvates, hydrates and/or salts thereof) and a carrier (a pharmaceutically acceptable carrier), further comprising a second chemotherapeutic and/or a second antiinflammatory agent.
  • the present compositions are useful for inhibiting abnormal cell growth or treating a hyperproliferative disorder in a mammal (e.g., human).
  • the present compositions are also useful for treating inflammatory diseases in a mammal (e.g., human).
  • the present invention includes a method of inhibiting abnormal cell growth or treating a hyperproliferative disorder in a mammal (e.g., human) comprising administering to said mammal a therapeutically effective amount of a compound of Formula I (and/or solvates and salts thereof) or a composition thereof, alone or in combination with a second chemotherapeutic agent.
  • the present invention includes a method of treating an inflammatory disease in a mammal (e.g., human) comprising administering to said mammal a therapeutically effective amount of a compound of Formula I (and/or solvates and salts thereof) or a composition thereof, alone or in combination with a second anti-inflammatory agent.
  • a mammal e.g., human
  • administering to said mammal a therapeutically effective amount of a compound of Formula I (and/or solvates and salts thereof) or a composition thereof, alone or in combination with a second anti-inflammatory agent.
  • the present invention includes a method of using the present compounds for in vitro, in situ, and in vivo diagnosis or treatment of mammalian cells, organisms, or associated pathological conditions.
  • alkyl refers to a saturated linear or branched-chain monovalent hydrocarbon radical of one to twelve carbon atoms.
  • alkyl groups include, but are not limited to, methyl (Me, -CH 3 ), ethyl (Et, -CH 2 CH 3 ), 1 -propyl (n-Pr, n- propyl, -CH 2 CH 2 CH 3 ), 2-propyl (i-Pr, i-propyl, -CH(CH 3 ) 2 ), 1 -butyl (n-Bu, n-butyl, - CH 2 CH 2 CH 3 ), 2 -methyl- 1 -propyl (i-Bu, i-butyl, -CH 2 CH(CH 3 ) 2 ), 2-butyl (s-Bu, s-butyl, - CH(CH 3 )CH 2 CH 3 ), 2-methyl-2-propyl (t-Bu, t-buty
  • alkynyl refers to a linear or branched monovalent hydrocarbon radical of two to twelve carbon atoms with at least one site of unsaturation, i.e., a carbon- carbon, sp triple bond. Examples include, but are not limited to, ethynyl (-C ⁇ CH), propynyl (propargyl, -CH 2 C ⁇ CH), and the like.
  • carrier refers to a monovalent non-aromatic, saturated or partially unsaturated ring having 3 to 12 carbon atoms as a monocyclic ring or 7 to 12 carbon atoms as a bicyclic ring.
  • Bicyclic carbocycles having 7 to 12 atoms can be arranged, for example, as a bicyclo [4,5], [5,5], [5,6] or [6,6] system, and bicyclic carbocycles having 9 or 10 ring atoms can be arranged as a bicyclo [5,6] or [6,6] system, or as bridged systems such as bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane and bicyclo[3.2.2]nonane.
  • monocyclic carbocycles include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-l-enyl, 1- cyclopent-2-enyl, l-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-l-enyl, l-cyclohex-2-enyl, 1- cyclohex-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, cyclododecyl, and the like.
  • Aryl means a monovalent aromatic hydrocarbon radical of 6-18 carbon atoms derived by the removal of one hydrogen atom from a single carbon atom of a parent aromatic ring system. Some aryl groups are represented in the exemplary structures as "Ar”. Aryl includes bicyclic radicals comprising an aromatic ring fused to a saturated, partially unsaturated ring, or aromatic carbocyclic or heterocyclic ring.
  • Typical aryl groups include, but are not limited to, radicals derived from benzene (phenyl), substituted benzenes, naphthalene, anthracene, indenyl, indanyl, 1 ,2-dihydronapthalene, 1,2,3,4-tetrahydronapthyl, and the like.
  • heterocycle refers to a saturated or a partially unsaturated (i.e., having one or more double and/or triple bonds within the ring) carbocyclic radical of 3 to 18 ring atoms in which at least one ring atom is a heteroatom selected from nitrogen, oxygen and sulfur, the remaining ring atoms being C, where one or more ring atoms is optionally substituted independently with one or more substituents described below.
  • a heterocycle may be a monocycle having 3 to 7 ring members (2 to 6 carbon atoms and 1 to 4 heteroatoms selected from N, O, P, and S) or a bicycle having 7 to 10 ring members (4 to 9 carbon atoms and 1 to 6 heteroatoms selected from N, O, P, and S), for example: a bicyclo [4,5], [5,5], [5,6], or [6,6] system.
  • Heterocycles are described in Paquette, Leo A.; "Principles of Modern Heterocyclic Chemistry" (W.A.
  • Heterocyclyl also includes radicals where heterocycle radicals are fused with a saturated, partially unsaturated ring, or aromatic carbocyclic or heterocyclic ring.
  • heterocyclic rings include, but are not limited to, pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino, thioxanyl, piperazinyl, homopiperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl, thiazepinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3- dioxolanyl, pyrazolinyl, dithianyl, dithiolany
  • heteroaryl refers to a monovalent aromatic radical of 5- or 6- membered rings, and includes fused ring systems (at least one of which is aromatic) of 5-18 atoms, containing one or more heteroatoms independently selected from nitrogen, oxygen, and sulfur.
  • heteroaryl groups are pyridinyl (including, for example, 2- hydroxypyridinyl), imidazolyl, imidazopyridinyl, pyrimidinyl (including, for example, 4- hydroxypyrimidinyl), pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, triazolyl, thiadiazolyl, furazanyl
  • the heterocycle or heteroaryl groups may be carbon (carbon-linked) or nitrogen (nitrogen-linked) attached where such is possible.
  • carbon bonded heterocycles or heteroaryls are bonded at position 2, 3, 4, 5, or 6 of a pyridine, position 3, 4, 5, or 6 of a pyridazine, position 2, 4, 5, or 6 of a pyrimidine, position 2, 3, 5, or 6 of a pyrazine, position 2, 3, 4, or 5 of a furan, tetrahydrofuran, thiofuran, thiophene, pyrrole or tetrahydropyrrole, position 2, 4, or 5 of an oxazole, imidazole or thiazole, position 3, 4, or 5 of an isoxazole, pyrazole, or isothiazole, position 2 or 3 of an aziridine, position 2, 3, or 4 of an azetidine, position 2, 3, 4, 5, 6, 7, or 8 of a quinoline or position 1, 3,
  • nitrogen bonded heterocycles or heteroaryls are bonded at position 1 of an aziridine, azetidine, pyrrole, pyrrolidine, 2- pyrroline, 3-pyrroline, imidazole, imidazolidine, 2-imidazoline, 3 -imidazoline, pyrazole, pyrazoline, 2-pyrazoline, 3-pyrazoline, piperidine, piperazine, indole, indoline, lH-indazole, position 2 of a isoindole, or isoindoline, position 4 of a morpholine, and position 9 of a carbazole, or ⁇ -carboline.
  • halo refers to F, Cl, Br or I.
  • heteroatoms present in heteroaryl or heterocyclcyl include the oxidized forms such as N + ⁇ O ⁇ , S(O) and S(O) 2 .
  • treat and treatment refer to both therapeutic treatment and prophylactic or preventative measures, wherein the object is to prevent or slow down (lessen) an undesired physiological change or disorder, such as the development or spread of cancer.
  • beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable.
  • Treatment can also mean prolonging survival as compared to expected survival if not receiving treatment.
  • Those in need of treatment include those already with the condition or disorder as well as those prone to have the condition or disorder or those in which the condition or disorder is to be prevented.
  • terapéuticaally effective amount means an amount of a compound of the present invention that (i) treats or prevents the particular disease, condition, or disorder, (ii) attenuates, ameliorates, or eliminates one or more symptoms of the particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of the particular disease, condition, or disorder described herein.
  • the therapeutically effective amount of the drug may reduce the number of cancer cells; reduce the tumor size; inhibit (i.e., slow to some extent and preferably stop) cancer cell infiltration into peripheral organs; inhibit (i.e., slow to some extent and preferably stop) tumor metastasis; inhibit, to some extent, tumor growth; and/or relieve to some extent one or more of the symptoms associated with the cancer.
  • the drug may prevent growth and/or kill existing cancer cells, it may be cytostatic and/or cytotoxic.
  • efficacy can be measured, for example, by assessing the time to disease progression (TTP) and/or determining the response rate (RR).
  • abnormal cell growth and “hyperproliferative disorder” are used interchangeably in this application.
  • Abnormal cell growth refers to cell growth that is independent of normal regulatory mechanisms (e.g., loss of contact inhibition).
  • tumor cells tumor cells
  • tumors tumor cells
  • any tumors that proliferate by receptor tyrosine kinases any tumors that proliferate by aberrant serine/threonine kinase activation
  • benign and malignant cells of other proliferative diseases in which aberrant serine/threonine kinase activation benign and malignant cells of other proliferative diseases in which aberrant serine/threonine kinase activation occurs.
  • cancer and “cancerous” refer to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth.
  • a “tumor” comprises one or more cancerous cells. Examples of cancer include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia or lymphoid malignancies.
  • squamous cell cancer e.g., epithelial squamous cell cancer
  • lung cancer including small- cell lung cancer, non-small cell lung cancer ("NSCLC"), adenocarcinoma of the lung and squamous carcinoma of the lung, cancer of the peritoneum, hepatocellular cancer, gastric or stomach cancer including gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial or uterine carcinoma, salivary gland carcinoma, kidney or renal cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma, anal carcinoma, penile carcinoma, acute leukemia, as well as head/brain and neck cancer.
  • NSCLC non-small cell lung cancer
  • adenocarcinoma of the lung and squamous carcinoma of the lung cancer of the peritoneum, hepatocellular cancer, gas
  • a "chemotherapeutic agent” is a chemical compound useful in the treatment of cancer.
  • chemotherapeutic agents include Erlotinib (TARCEVA®, Genentech/OSI Pharm.), Bortezomib (VELCADE®, Millennium Pharm.), Fulvestrant (FASLODEX®, AstraZeneca), Sutent (SUl 1248, Pfizer), Letrozole (FEMARA®, Novartis), Imatinib mesylate (GLEEVEC®, Novartis), PTK787/ZK 222584 (Novartis), Oxaliplatin (Eloxatin®, Sanofi), 5-FU (5-fluorouracil), Leucovorin, Rapamycin (Sirolimus, RAPAMUNE®, Wyeth), Lapatinib (TYKERB®, GSK572016, Glaxo Smith Kline), Lonafarnib (SCH 66336), Sorafenib (BAY43-9006, Bayer Lab
  • dynemicin including dynemicin A; bisphosphonates, such as clodronate; an esperamicin; as well as neocarzinostatin chromophore and related chromoprotein enediyne antibiotic chromophores), aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, carabicin, carminomycin, carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin, 6- diazo-5-oxo-L-norleucine, ADRIAMYCIN® (doxorubicin), morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin,
  • chemotherapeutic agent also included in the definition of "chemotherapeutic agent” are: (i) anti- hormonal agents that act to regulate or inhibit hormone action on tumors such as anti- estrogens and selective estrogen receptor modulators (SERMs), including, for example, tamoxifen (including NOLVADEX®; tamoxifen citrate), raloxifene, droloxifene, A- hydroxytamoxifen, trioxifene, keoxifene, LYl 17018, onapristone, and FARESTON® (toremifine citrate); (ii) aromatase inhibitors that inhibit the enzyme aromatase, which regulates estrogen production in the adrenal glands, such as, for example, 4(5)-imidazoles, aminoglutethimide, MEGASE® (megestrol acetate), AROMASIN® (exemestane; Pfizer), formestanie, fadrozole, RTVISOR® (vorozole), FEMARA
  • anti-angiogenic agents include MMP -2 (matrix- metalloproteinase 2) inhibitors, MMP-9 (matrix-metalloproteinase 9) inhibitors, COX-II (cyclooxygenase II) inhibitors, and VEGF receptor tyrosine kinase inhibitors.
  • VEGF receptor tyrosine kinase inhibitors include 4-(4-bromo-2-fluoroanilino)-6-methoxy-7- (l-methylpiperidin-4-ylmethoxy)quinazoline (ZD6474; Example 2 within WO 01/32651), 4- (4-fluoro-2-methylindol-5-yloxy)-6-methoxy-7-(3-pyrrolidin-l-ylpropoxy)- quinazoline (AZD2171; Example 240 within WO 00/47212), vatalanib (PTK787; WO 98/35985) and SUl 1248 (sunitinib; WO 01/60814), and compounds such as those disclosed in PCT Publication Nos.
  • chemotherapeutic agents that can be used in combination with the present compounds include inhibitors of PI3K (phosphoinositide-3 kinase), such as those reported in Yaguchi et al (2006) Jour, of the Nat. Cancer Inst.
  • PI3K phosphoinositide-3 kinase
  • inflammatory diseases includes, but not limited to, rheumatoid arthritis, atherosclerosis, congestive hear failure, inflammatory bowel disease (including, but not limited to, Crohn's disease and ulcerative colitis), chronic obstructive pulmonary disease in the lung, f ⁇ brotic disease in the liver and kidney, Crohn's disease, skin diseases such as psoriasis, eczema and scleroderma, osteoarthritis, multiple sclerosis, asthma, diseases and disorders related to diabetic complications, f ⁇ brotic organ failure in organs such as lung, liver, kidney, and inflammatory complications of the cardiovascular system such as acute coronary syndrome.
  • inflammatory diseases includes, but not limited to, rheumatoid arthritis, atherosclerosis, congestive hear failure, inflammatory bowel disease (including, but not limited to, Crohn's disease and ulcerative colitis), chronic obstructive pulmonary disease in the lung, f ⁇ brotic disease in the liver and kidney, Crohn's disease,
  • anti-inflammatory agent is a compound useful in the treatment of inflammation.
  • anti-inflammatory agents include injectable protein therapeutics such as Enbrel®, Remicade®, Humira® and Kineret®.
  • Other examples of anti-inflammatory agents include non-steroidal anti-inflammatory agents (NSAIDs), such as ibuprofen or aspirin (which reduce swelling and alleviate pain); disease-modifying anti-rheumatic drugs (DMARDs) such as methotrexate; 5-aminosalicylates (sulfasalazine and the sulfa-free agents); corticosteroids; immunomodulators such as 6-mercaptoputine ("6-MP"), azathioprine (“AZA”), cyclosporins, and biological response modifiers such as Remicade.RTM.
  • NSAIDs non-steroidal anti-inflammatory agents
  • DMARDs disease-modifying anti-rheumatic drugs
  • DMARDs such as methotrexate
  • 5-aminosalicylates s
  • infliximab and Enbrel.RTM. (etanercept); fibroblast growth factors; platelet derived growth factors; enzyme blockers such as Arava.RTM. (leflunomide); and/or a cartilage protecting agent such as hyaluronic acid, glucosamine, and chondroitin.
  • prodrug refers to a precursor or derivative form of a compound of the invention that is capable of being enzymatically or hydrolytically activated or converted into the more active parent form. See, e.g., Wilman, "Prodrugs in Cancer Chemotherapy” Biochemical Society Transactions, 14, pp. 375-382, 615th Meeting Harbor (1986) and Stella et al., “Prodrugs: A Chemical Approach to Targeted Drug Delivery,” Directed Drug Delivery, Borchardt et al., (ed.), pp. 247-267, Humana Press (1985).
  • the prodrugs of this invention include, but are not limited to, ester-containing prodrugs, phosphate-containing prodrugs, thiophosphate-containing prodrugs, sulfate- containing prodrugs, peptide-containing prodrugs, D-amino acid-modified prodrugs, glycosylated prodrugs, ⁇ -lactam-containing prodrugs, optionally substituted phenoxyacetamide-containing prodrugs, optionally substituted phenylacetamide-containing prodrugs, 5-fluorocytosine and other 5-fluorouridine prodrugs which can be converted into the more active cytotoxic free drug.
  • cytotoxic drugs that can be derivatized into a prodrug form for use in this invention include, but are not limited to, compounds of the invention and chemotherapeutic agents such as described above.
  • a "metabolite” is a product produced through metabolism in the body of a specified compound or salt thereof. Metabolites of a compound may be identified using routine techniques known in the art and their activities determined using tests such as those described herein. Such products may result for example from the oxidation, hydroxylation, reduction, hydrolysis, amidation, deamidation, esterif ⁇ cation, deesterif ⁇ cation, enzymatic cleavage, and the like, of the administered compound. Accordingly, the invention includes metabolites of compounds of the invention, including compounds produced by a process comprising contacting a compound of this invention with a mammal for a period of time sufficient to yield a metabolic product thereof.
  • a “liposome” is a small vesicle composed of various types of lipids, phospholipids and/or surfactant which is useful for delivery of a drug (such as MEK kinase inhibitors disclosed herein and, optionally, a chemotherapeutic agent) to a mammal.
  • a drug such as MEK kinase inhibitors disclosed herein and, optionally, a chemotherapeutic agent
  • the components of the liposome are commonly arranged in a bilayer formation, similar to the lipid arrangement of biological membranes.
  • package insert is used to refer to instructions customarily included in commercial packages of therapeutic products, that contain information about the indications, usage, dosage, administration, contraindications and/or warnings concerning the use of such therapeutic products.
  • chiral refers to molecules which have the property of non-superimposability of the mirror image partner, while the term “achiral” refers to molecules which are superimposable on their mirror image partner.
  • stereoisomer refers to compounds which have identical chemical constitution and connectivity, but different orientations of their atoms in space that cannot be interconverted by rotation about single bonds.
  • Diastereomer refers to a stereoisomer with two or more centers of chirality and whose molecules are not mirror images of one another. Diastereomers have different physical properties, e.g. melting points, boiling points, spectral properties, and reactivities.
  • Mixtures of diastereomers may separate under high resolution analytical procedures such as crystallization, electrophoresis and chromatography.
  • Enantiomers refer to two stereoisomers of a compound which are non- superimposable mirror images of one another. [0046] Stereochemical definitions and conventions used herein generally follow S. P.
  • the compounds of the invention may contain asymmetric or chiral centers, and therefore exist in different stereoisomeric forms. It is intended that all stereoisomeric forms of the compounds of the invention, including but not limited to, diastereomers, enantiomers and atropisomers, as well as mixtures thereof such as racemic mixtures, form part of the present invention.
  • Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane-polarized light.
  • the prefixes D and L, or R and S are used to denote the absolute configuration of the molecule about its chiral center(s).
  • the prefixes d and 1 or (+) and (-) are employed to designate the sign of rotation of plane-polarized light by the compound, with (-) or 1 meaning that the compound is levorotatory.
  • a compound prefixed with (+) or d is dextrorotatory.
  • these stereoisomers are identical except that they are mirror images of one another.
  • a specific stereoisomer may also be referred to as an enantiomer, and a mixture of such isomers is often called an enantiomeric mixture.
  • a 50:50 mixture of enantiomers is referred to as a racemic mixture or a racemate, which may occur where there has been no stereoselection or stereospecificity in a chemical reaction or process.
  • racemic mixture and “racemate” refer to an equimolar mixture of two enantiomeric species, devoid of optical activity.
  • tautomer or “tautomeric form” refers to structural isomers of different energies which are interconvertible via a low energy barrier.
  • proton tautomers also known as prototropic tautomers
  • Valence tautomers include interconversions by reorganization of some of the bonding electrons.
  • pharmaceutically acceptable salt refers to pharmaceutically acceptable organic or inorganic salts of a compound of the invention.
  • Exemplary salts include, but are not limited, to sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate "mesylate", ethanesulfonate, benzenesulfonate, />-toluenesulfonate, pamoate (i.e., l,l'-methylene-bis -(2-hydroxy-3 -naphthoate)) salts, alkali metal (e.g., sodium and potassium) salts
  • a pharmaceutically acceptable salt may involve the inclusion of another molecule such as an acetate ion, a succinate ion or other counter ion.
  • the counter ion may be any organic or inorganic moiety that stabilizes the charge on the parent compound.
  • a pharmaceutically acceptable salt may have more than one charged atom in its structure. Instances where multiple charged atoms are part of the pharmaceutically acceptable salt can have multiple counter ions. Hence, a pharmaceutically acceptable salt can have one or more charged atoms and/or one or more counter ion.
  • the desired pharmaceutically acceptable salt may be prepared by any suitable method available in the art, for example, treatment of the free base with an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, methanesulfonic acid, phosphoric acid and the like, or with an organic acid, such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, a pyranosidyl acid, such as glucuronic acid or galacturonic acid, an alpha hydroxy acid, such as citric acid or tartaric acid, an amino acid, such as aspartic acid or glutamic acid, an aromatic acid, such as benzoic acid or cinnamic acid, a sulfonic acid, such as p-toluenesulfonic acid or ethanes
  • an inorganic acid such as hydrochloric acid
  • the desired pharmaceutically acceptable salt may be prepared by any suitable method, for example, treatment of the free acid with an inorganic or organic base, such as an amine (primary, secondary or tertiary), an alkali metal hydroxide or alkaline earth metal hydroxide, or the like.
  • an inorganic or organic base such as an amine (primary, secondary or tertiary), an alkali metal hydroxide or alkaline earth metal hydroxide, or the like.
  • Suitable salts include, but are not limited to, organic salts derived from amino acids, such as glycine and arginine, ammonia, primary, secondary, and tertiary amines, and cyclic amines, such as piperidine, morpholine and piperazine, and inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum and lithium.
  • organic salts derived from amino acids such as glycine and arginine, ammonia, primary, secondary, and tertiary amines, and cyclic amines, such as piperidine, morpholine and piperazine
  • inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum and lithium.
  • a “solvate” refers to an association or complex of one or more solvent molecules and a compound of the invention.
  • solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanolamine.
  • hydrate refers to the complex where the solvent molecule is water.
  • protecting group refers to a substituent that is commonly employed to block or protect a particular functionality while reacting other functional groups on the compound.
  • an “amino-protecting group” is a substituent attached to an amino group that blocks or protects the amino functionality in the compound.
  • Suitable amino- protecting groups include acetyl, trifluoroacetyl, t-butoxycarbonyl (BOC), benzyloxycarbonyl (CBZ) and 9-fluorenylmethylenoxycarbonyl (Fmoc).
  • a "hydroxy-protecting group” refers to a substituent of a hydroxy group that blocks or protects the hydroxy functionality. Suitable protecting groups include acetyl and silyl.
  • a “carboxy-protecting group” refers to a substituent of the carboxy group that blocks or protects the carboxy functionality.
  • Common carboxy-protecting groups include phenylsulfonylethyl, cyanoethyl, 2-(trimethylsilyl)ethyl, 2-(trimethylsilyl)ethoxymethyl, 2-(p-toluenesulfonyl)ethyl, 2-(p- nitrophenylsulfenyl)ethyl, 2-(diphenylphosphino)-ethyl, nitroethyl and the like.
  • protecting groups and their use see T. W. Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, New York, 1991.
  • the present invention provides azabenzofuranyl compounds of Formula I as described above useful as kinase inhibitors, particularly useful as MEK kinase inhibitors.
  • the present invention includes compounds of Formulae I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, I-i, II-a, II-b, II-c, II-d, II-e, II-f, II-g, II-h, II-i, III-a, III-b, III-c, III-d, III-e, III-f, III-g, III-h, and III-i, and all other variables are as defined in Formula I.
  • compounds are of Formulae I-b, I- f, I-g, I-h, II-b, II-f, II-g, II-h, III-b, III-f, III-g and III-h, and all other variables are as defined in Formula I.
  • R 1 is H, halo, CN, CF 3 , -NR 11 R 12 ,
  • R 1 is H, halo, CN, CF 3 ,
  • III-d III-d, III-f, or III-g.
  • R 1 is H, Cl, CN, CF 3 , methyl
  • R 1 is H; and all other variables are as defined in Formula I, I-a, I-b, I-d, I-f, I-g, II-a, II-b, II-d, II-f, II-g, III-a, III-b,
  • III-d III-d, III-f, or III-g.
  • R 2 is H, halo, CN, CF 3 , -NR 11 R 12 ,
  • R is H, halo, CN, CF 3 ,
  • III-d, III-e, or III-i or as defined above.
  • R 2 is H, Cl, CN, CF 3 , methyl
  • R 3 is H, halo, CN, CF 3 , -NR 11 R 12 ,
  • R 3 is H, halo, CF 3 , Ci-C 6 alkyl; and all other variables are as defined in Formula I, I-a, I-c, I-d, I-e, I-i, II-a, II-c, II-d,
  • III-e II-e, II-i, III-a, III-c, III-d, III-e, or III-i, or as defined above.
  • R 3 is H, F, CF 3 , or methyl; and all other variables are as defined in Formula I, I-a, I-c, I-d, I-e, I-i, II-a, II-c, II-d, II-e, II- i, III-a, III-c, III-d, III-e, or III-i, or as defined above.
  • R is H, F, Cl, CF 3 , methyl or
  • R 4 is H, halo, CN, CF 3 , -NR 11 R 12 ,
  • R 4 is H, halo, CN, CF 3 ,
  • R 11 and R 12 are independently H or Ci-C 6 alkyl, -OR 11 wherein R 11 is H or Ci -C 6 alkyl, or -SR 11 wherein R 11 is H or Ci -C 6 alkyl; and all other variables are as defined in Formula I, I-a, I-b, I-c, I-e, I-g, I-h, II-a, II-b, II-c, II-e, II-g, II-h,
  • R 4 is H, Br, Cl, CN, CF 3 ,
  • R 4 is H, Br, Cl, CN, CF 3 ,
  • R 4 is halo, -OH, or Ci-C 6 alkyl optionally substituted by halo; and all other variables are as defined in Formula I, I-a, I- b, I-c, I-e, I-g, I-h, II-a, II-b, II-c, II-e, II-g, II-h, III-a, III-b, III-c, III-e, III-g, or III-h, or as defined above.
  • R 4 is independently Cl, Br,
  • R 5 is H or Ci-C 6 alkyl; and all other variables are as defined in Formula I, I-a to I-i, or II-a to II-i, or as defined above.
  • R 5 is H or methyl; and all other variables are as defined in Formula I, I-a to I-i, or II-a to II-i, or as defined above.
  • R 5 is H; and all other variables are as defined in Formula I, I-a to I-i, or II-a to II-i, or as defined above.
  • R 5 is methyl; and all other variables are as defined in Formula I, I-a to I-i, or II-a to II-i, or as defined above.
  • R 6 is H or Ci-C 6 alkyl; and all other variables are as defined in Formula I, I-a to I-i, II-a to II-i, or III-a to III-i, or as defined above.
  • R 6 is H or methyl; and all other variables are as defined in Formula I, I-a to I-i, II-a to II-i, or III-a to III-i, or as defined above.
  • R 6 is H; and all other variables are as defined in Formula I, I-a to I-i, II-a to II-i, or III-a to III-i, or as defined above.
  • R 6 is methyl; and all other variables are as defined in Formula I, I-a to I-i, II-a to II-i, or III-a to III-i, or as defined above.
  • X 1 is OR 11 (i.e., Formula II-a to II- i); and all other variables are as defined in Formula I or I-a to I-i; or as defined above.
  • X 1 is OR 11 wherein R 11 is H; and all other variables are as defined in Formula I or I-a to I-i; or as defined above.
  • X 1 is OR 11 wherein R 11 is Ci-
  • X 1 is:
  • X 1 is ; and all other variables are as defined in Formula I or I-a to I-i, or as defined above.
  • X 1 is
  • X 1 is OR 11 wherein R 11 is heterocyclyl (e.g., 4- to 6-membered heterocyclyl) optionally substituted with one or more groups independently selected from halo, CN, CF 3 , -OCF 3 , -NO 2 , oxo, -Si(Ci-Co alkyl),
  • X 1 is .
  • X 1 is N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoe
  • X 1 is R 11
  • X 1 is R 11
  • W is:
  • X 1 is R 11
  • W is:
  • X 1 is -OR 11 , and -OR 11 of X 1 and
  • X 1 is -OR 11
  • X 1 is -OR 11
  • X 1 is R 11 ; and all other variables are as defined in Formula I, I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, or I-i, or as defined above.
  • X 1 is R 11 wherein R 11 is H; and all other variables are as defined in Formula I, I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, or I-i, or as defined above.
  • X 1 is R 11 wherein R 11 is Ci-
  • X 1 is N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoe
  • X 1 is -S(O) 2 R 11 , and all other variables are as defined in Formula I, I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, or I-i, or as defined above.
  • X 1 is -S(O) 2 R 11 wherein R 11 is H or methyl; and all other variables are as defined in Formula I, I-a, I-b, I-c, I-d, I-e, I-f, I- g, I-h, or I-i, or as defined above.
  • W is -OR 11 (i.e., Formula III-a, III- b, III-c, Ill-d, Ill-e, III-f, III-g, III-h, or III-i) wherein R 11 of W is H or Ci-Ci 2 alkyl; and all other variables are as defined above.
  • W is -OR 11 (i.e., Formula III- a, Ill-b, III-c, III-d, III-e, III-f, III-g, III-h, or III-i) wherein R 11 of W is H; and all other variables are as defined above.
  • W is -OR 11 (i.e., Formula III- a, Ill-b, III-c, III-d, III-e, III-f, III-g, III-h, or III-i) wherein R 11 of W is Ci-C 6 alkyl; and all other variables are as defined above.
  • X 2 is
  • X 2 is
  • X 2 is ; and all other variables are as defined in Formula I, I-a to I-i, II-a to II-i, or III- a to III-i, or as defined above.
  • X 2 is C ⁇ -Cio aryl substituted with Ci -C 4 alkyl; and all other variables are as defined in Formula I, I-a to I-i, II-a to II-i, or III-a to III-i, or as defined above. [00124] In another embodiment of the present invention, X 2 is
  • X 2 is
  • X is C 4 -C 6 carb
  • anc ⁇ an other variables are as defined in Formula I, I-a to I-i, II-a to II-i, or III-a to III-i, or as defined above.
  • Another embodiment of the present invention includes compounds described in EXAMPLES 5-159 and compounds below:
  • the present compounds are prepared according to the procedures described below in the schemes and examples or by methods known in the art.
  • the starting materials and various intermediates may be obtained from commercial sources, prepared from commercially available compounds, or prepared using well known synthetic methods (for example, those described in WO02/06213, WO 03/077855 and WO03/077914).
  • 5-azabenzofurans of Formula (I-b), (II-b) or (III-b) may be prepared using the synthetic routes outlined in Schemes 1, 2 and 3.
  • Compounds of formula (IV) may be prepared using published methods described in the literature. They may be reacted with methylglycolate or ethylglycolate in the presence of a base, such as sodium hydride, in a suitable solvent, such as NN- dimethylformamide or 1 ,2-dimethoxyethane, at a temperature of from -5O 0 C to room temperature, to obtain compounds of formula (VI).
  • a base such as sodium hydride
  • a suitable solvent such as NN- dimethylformamide or 1 ,2-dimethoxyethane
  • Compounds of formula (VI) may be converted to compounds of formula (VII) by reaction with a halogenating agent such as phosphorus oxybromide, neat or in a suitable solvent such as toluene, at a temperature of from room temperature to 14O 0 C.
  • a halogenating agent such as phosphorus oxybromide
  • a suitable solvent such as toluene
  • compounds of formula (VI) may be reacted with nonafluorobutane sulphonyl fluoride in the presence of a base such as diisopropylethylamine and a catalyst such as NN-dimethyl-4- aminopyridine, in a solvent such as dichloromethane at room temperature, with N- phenyltrifluoromethanesulfonimide in the presence of a base such as diisopropylethylamine, in a suitable solvent such as 1,2-dimethoxyethane at a temperature from room temperature to the reflux temperature of the solvent.
  • a base such as diisopropylethylamine and a catalyst such as NN-dimethyl-4- aminopyridine
  • a solvent such as dichloromethane at room temperature
  • N- phenyltrifluoromethanesulfonimide in the presence of a base such as diisopropylethylamine, in a suitable solvent such as 1,2-dimethoxye
  • compounds of formula (VI) may be treated with trifluoromethanesulphonic acid anhydride in the presence of a base such as pyridine in a solvent such as dichloromethane at a temperature of from -2O 0 C to ambient temperature.
  • a base such as pyridine
  • a solvent such as dichloromethane
  • VII by reaction with an aniline (incorporating appropriate substituents Rl), in the presence of a catalyst such as tris(dibenzylideneacetone)dipalladium (0) or palladium acetate, a base such as potassium phosphate, sodium tert-butoxide, l,8-diazabicyclo[5.4.1]undec-7-ene or cesium carbonate, a ligand such as 9,9'-dimethyl-4,5-bis(diphenylphosphino)xanthene, 2,2'- bis(diphenylphosphino)-l,l '-binaphthyl, 2-dicyclohexylphosphino-2'-(NN- dimethylamino)biphenyl, 2-dicyclohexylphosphino-2',6'-(dimethoxy)biphenyl or tri-butyl- phosphine in a suitable solvent such as toluene, 1,2-dimethoxy
  • compounds of formula (VIII) can be obtained from compounds of formula (VI) by reaction with compounds of formula (IX) (prepared using published methods described in the literature), in a suitable solvent such as toluene or 1,2- dimethoxyethane, at a temperature of from room temperature to the reflux temperature of the solvent, or under microwave irradiation at a temperature of from 100 0 C to 18O 0 C.
  • compounds of formula (X) can be obtained from compounds of formula
  • compounds of formula (XI) can be obtained directly from compounds of formula (VIII) by reaction with an amine or hydroxylamine D ⁇ HR in the presence of a Lewis acid such as trimethyl aluminium in a solvent such as DCM, at a temperature of from room temperature up to reflux temperature.
  • a Lewis acid such as trimethyl aluminium in a solvent such as DCM
  • R' is Me or Et Base, Solvent (XVI) or lower alkyl
  • Compounds of formula (XIII) may be prepared using published methods described in the literature.
  • Compounds of general formula (XIV) can be prepared from compounds of formula (XIII) using methods described above for the preparation of compounds of formula (VI) from compounds of formula (IV).
  • compounds of formula (XIV) by reaction with compounds of formula (XV) (incorporating appropriate substituents Rl), using methods described above for the preparation of compounds of formula (VIII) from compounds of formula (VI).
  • compounds of formula (VIII) may be obtained from compounds of formula (XIV) by reaction with compounds of formula (XVI) (incorporating appropriate substituents Rl), in the presence of a base such as sodium hydride or lithium hexamethyldisilazane, in a suitable solvent such as tetrahydrofuran or N, N- dimethylformamide, at a temperature of from room temperature to 15O 0 C.
  • a base such as sodium hydride or lithium hexamethyldisilazane
  • a suitable solvent such as tetrahydrofuran or N, N- dimethylformamide
  • Compounds of formula (XVII) can be coupled to amines such as 2-amino-2- methyl-1-propanol using methods described above for the preparation of compounds of formula (XI) from compounds of formula (X), followed by reaction with an agent such as thionyl chloride or phosphorus oxychloride, neat or in a suitable solvent such as dichloromethane, chloroform or diethyl ether, at a temperature of from room temperature to reflux of the solvent, to afford compounds of formula (XVIII).
  • an agent such as thionyl chloride or phosphorus oxychloride, neat or in a suitable solvent such as dichloromethane, chloroform or diethyl ether, at a temperature of from room temperature to reflux of the solvent, to afford compounds of formula (XVIII).
  • Compounds of formula (XIX) may be obtained from compounds of formula
  • XVIII by reaction with an aniline (incorporating appropriate substituents Rl), in the presence of a catalyst such as tris(dibenzylideneacetone)dipalladium (0) or palladium acetate, a base such as potassium phosphate, sodium tert-butoxide, l,8-diazabicyclo[5.4.1]undec-7- ene or cesium carbonate, a ligand such as 9,9'-dimethyl-4,5-bis(diphenylphosphino)xanthene, 2,2'-bis(diphenylphosphino)-l,l '-binaphthyl, 2-dicyclohexylphosphino-2'-(N,N- dimethylamino)biphenyl, 2-dicyclohexylphosphino-2',6'-(dimethoxy)biphenyl or tri-butyl- phosphine in a suitable solvent such as toluene, 1, 2
  • compounds of formula (XIX) may be obtained from compounds of formula (XVIII) by reaction with anilines (incorporating appropriate substituents Rl), in the presence of a base such as sodium hydride or lithium hexamethyldisilazane, in a suitable solvent such as tetrahydrofuran or N,N-dimethylformamide, at a temperature of from room temperature to 15O 0 C.
  • anilines incorporating appropriate substituents Rl
  • a base such as sodium hydride or lithium hexamethyldisilazane
  • a suitable solvent such as tetrahydrofuran or N,N-dimethylformamide
  • 6-Azabenzofurans of Formula I-c, II-c or III-c may be prepared using the synthetic routes outlined in Scheme 4.
  • Compounds of formula (XX) may be prepared using published methods described in the literature. They may be reacted with methylglycolate or ethylglycolate in the presence of a phosphine such as triphenyl phosphine, an alkyl-azodicarboxylate such as diethyl azodicarboxylate or diisopropyl azodicarboxylate, in an aprotic solvent, such as tetrahydrofuran or diethyl ether, at a temperature of from room temperature to reflux of the solvent, to obtain compounds of formula (XXI).
  • a phosphine such as triphenyl phosphine
  • an alkyl-azodicarboxylate such as diethyl azodicarboxylate or diisopropyl azodicarboxylate
  • an aprotic solvent such as tetrahydrofuran or diethyl ether
  • compounds of formula (VI) may be treated with trifluoromethanesulphonic acid anhydride in the presence of a base such as pyridine in a solvent such as dichloromethane at a temperature of from - 2O 0 C to ambient temperature.
  • a base such as pyridine
  • a solvent such as dichloromethane
  • Compounds of formula (XXIV) may be obtained from compounds of formula (XXIII) by reaction with an aniline (incorporating appropriate substituents Rl), in the presence of a catalyst such as tris(dibenzylideneacetone)dipalladium (0) or palladium acetate, a base such as potassium phosphate, sodium tert-butoxide, l,8-diazabicyclo[5.4.1]undec-7-ene or cesium carbonate, a ligand such as 9,9'-dimethyl-4,5-bis(diphenylphosphino)xanthene, 2,2'- bis(diphenylphosphino)-l,l '-binaphthyl, 2-dicyclohexylphosphino-2'-(NN- dimethylamino)biphenyl, 2-dicyclohexylphosphino-2',6'-(dimethoxy)biphenyl or tri-butyl- phosphine
  • compounds of formula (XXIV) can be obtained from compounds of formula (XXII) by reaction with compounds of formula (IX) (prepared using published methods described in the literature), in a suitable solvent such as toluene or 1,2- dimethoxyethane, at a temperature from room temperature to the reflux temperature of the solvent, or under microwave irradiation at a temperature of from 100 0 C to 18O 0 C.
  • a suitable solvent such as toluene or 1,2- dimethoxyethane
  • compounds of formula (XXVI) can be obtained directly from compounds of formula (XXIV) by reaction with an amine or hydroxylamine D ⁇ HR in the presence of a Lewis acid such as trimethyl aluminium, in a solvent such as DCM, at a temperature of from room temperature up to reflux temperature.
  • a Lewis acid such as trimethyl aluminium
  • Furo[2,3-d]pyrimidines of Formula I-f, II-f or III-f may be prepared using the synthetic routes outlined in Scheme 5.
  • Compounds of formula (XXVIII) may be prepared according to methods described in the literature. They may reacted with a halogenating agent such as phosphorus oxychloride, neat or in a suitable solvent such as toluene, at a temperature from room temperature to reflux, to provide compounds of formula (XXIX).
  • a halogenating agent such as phosphorus oxychloride
  • a suitable solvent such as toluene
  • Compounds of formula (L) may be prepared according to methods described in the literature. Compounds of formula (LVI) may be obtained from compounds of formula (L) using similar methods to the ones described for the preparation of compounds of formula (XI) from compounds of formula (IV), as shown in Scheme 1. Alternatively, compounds of formula (LIV) may be prepared according to Scheme 7.
  • a 1 N
  • Hydroxylamines of formula (XII) may be prepared using methods described in the literature or the synthetic route outlined in Scheme 8.
  • Primary or secondary alcohols of general formula (XXXVII) may be prepared using methods described in the literature. They may be reacted with N-hydroxy phthalimide using a phosphine and coupling reagent such as diethyl azodicarboxylate to provide compounds of general formula (XXXVIII). Compounds of general formula (XXXVIII) may be deprotected using hydrazine or methyl hydrazine to provide hydroxylamines of general formula (XII-a).
  • Compounds of formula (XII-a) may be further modified by reductive amination with aldehydes or ketones using a reducing agent such as sodium triacetoxy borohydride, sodium cyanoborohydride, or borane-pyridine in a solvent such as dichloroethane at a temperature of from ambient temperature to reflux.
  • a reducing agent such as sodium triacetoxy borohydride, sodium cyanoborohydride, or borane-pyridine
  • a solvent such as dichloroethane
  • compounds of formula (XII-a) may be further modified by alkylation with an alkyl halide in the presence of a base such as triethylamine, in a solvent such as dichloromethane, to provide hydroxylamines of general formula (XII-b).
  • R 1 is an optional substituent group
  • n 0- 4
  • Substituted 4-chloro-nitro benzene may be reacted with hexamethyldisilane in a solvent such as xylene using a catalyst such as tetrakis(triphenylphosphine)palladium at a temperature of from room temperature to reflux.
  • the nitro group may be reduced using methods described in the literature such as reaction under an atmosphere of hydrogen at a pressure of from 1 to 5 atmospheres in the presence of a catalyst such as palladium on carbon and in a solvent such as ethanol or ethyl acetate at room temperature.
  • Trifluoromethanesulfonyl esters of general formula (XL) used in cross- coupling reactions described above may be prepared by using methods described in the literature or according to Scheme 10.
  • n 0- ⁇
  • Halo phenols of general structure (XLI) may be reacted with two equivalents of alkylithium reagents such as n-butyl lithium in a solvent such as THF, followed by quenching with trialkylsilyl halide such as trimethylsilyl chloride to give trialkylsilyl phenols (XLII).
  • Trialkylsilyl phenols may be further reacted using literature procedures to give trifluoromethane sulfonates or nonaflates of general structure (XL).
  • compounds of formula (I), (II), (III) or any intermediates used in their preparation may be further derivatised by one or more standard synthetic methods employing substitution, oxidation, reduction, or cleavage reactions.
  • Particular substitution approaches include conventional alkylation, arylation, heteroarylation, acylation, sulfonylation, halogenation, nitration, formylation and coupling procedures.
  • aryl bromide or chloride groups may be converted to aryl iodides using a Finkelstein reaction employing an iodide source such as sodium iodide, a catalyst such as copper iodide and a ligand such as trans-N,N'-dimethyl-l,2-cyclohexane diamine in a solvent such as 1,4-dioxane and heating the reaction mixture at reflux temperature.
  • an iodide source such as sodium iodide
  • a catalyst such as copper iodide
  • a ligand such as trans-N,N'-dimethyl-l,2-cyclohexane diamine
  • Aryl trialkylsilanes may be converted to aryl iodides by treating the silane with an iodide source such as iodine monochloride in a solvent such as dichloromethane with or without Lewis acid such as silver tetrafluoroborate at a temperature from -4O 0 C to reflux.
  • an iodide source such as iodine monochloride in a solvent such as dichloromethane
  • Lewis acid such as silver tetrafluoroborate
  • primary amine (-NH 2 ) groups may be alkylated using a reductive alkylation process employing an aldehyde or a ketone and a borohydride, for example sodium triacetoxyborohydride or sodium cyanoborohydride, in a solvent such as a halogenated hydrocarbon, for example 1,2-dichloroethane, or an alcohol such as ethanol, where necessary in the presence of an acid such as acetic acid at around ambient temperature.
  • Secondary amine (-NH-) groups may be similarly alkylated employing an aldehyde.
  • primary amine or secondary amine groups may be converted into amide groups (-NHCOR' or -NRCOR') by acylation.
  • Acylation may be achieved by reaction with an appropriate acid chloride in the presence of a base, such as triethylamine, in a suitable solvent, such as dichloromethane, or by reaction with an appropriate carboxylic acid in the presence of a suitable coupling agent such HATU (O-(7- azabenzotriazol-l-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate) in a suitable solvent such as dichloromethane.
  • HATU O-(7- azabenzotriazol-l-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate
  • amine groups may be converted into sulphonamide groups (-NHSO 2 R' or -NR"SO 2 R') groups by reaction with an appropriate sulphonyl chloride in the presence of a suitable base, such as triethylamine, in a suitable solvent such as dichloromethane.
  • Primary or secondary amine groups can be converted into urea groups (-NHCONR'R" or -NRCONR'R”) by reaction with an appropriate isocyanate in the presence of a suitable base such as triethylamine, in a suitable solvent, such as dichloromethane.
  • An amine (-NH 2 ) may be obtained by reduction of a nitro (-NO 2 ) group, for example by catalytic hydrogenation, using for example hydrogen in the presence of a metal catalyst, for example palladium on a support such as carbon in a solvent such as ethyl acetate or an alcohol e.g. methanol.
  • a metal catalyst for example palladium on a support such as carbon in a solvent such as ethyl acetate or an alcohol e.g. methanol.
  • the transformation may be carried out by chemical reduction using for example a metal, e.g. tin or iron, in the presence of an acid such as hydrochloric acid.
  • amine (-CH 2 NH 2 ) groups may be obtained by reduction of nitriles (-CN), for example by catalytic hydrogenation using for example hydrogen in the presence of a metal catalyst, for example palladium on a support such as carbon, or Raney nickel, in a solvent such as an ether e.g. a cyclic ether such as tetrahydrofuran, at a temperature from -78°C to the reflux temperature of the solvent.
  • a metal catalyst for example palladium on a support such as carbon, or Raney nickel
  • Aldehyde groups (-CHO) may be converted to amine groups (-CH 2 NR 5 R")) by reductive amination employing an amine and a borohydride, for example sodium triacetoxyborohydride or sodium cyanoborohydride, in a solvent such as a halogenated hydrocarbon, for example dichloromethane, or an alcohol such as ethanol, where necessary in the presence of an acid such as acetic acid at around ambient temperature.
  • a borohydride for example sodium triacetoxyborohydride or sodium cyanoborohydride
  • a solvent such as a halogenated hydrocarbon, for example dichloromethane, or an alcohol such as ethanol
  • aldehyde groups may be converted into alkenyl groups (-
  • Aldehyde groups may be obtained by reduction of ester groups (such as -
  • aldehyde groups may be obtained by the oxidation of alcohol groups using any suitable oxidising agent known to those skilled in the art.
  • Ester groups (-CO 2 R') may be converted into the corresponding acid group (-
  • acid-catalysed hydrolysis can be achieved for example by treatment with an organic acid such as trifluoroacetic acid in an aqueous solvent, or by treatment with an inorganic acid such as hydrochloric acid in an aqueous solvent.
  • Carboxylic acid groups (-CO 2 H) may be converted into amides (CONHR' or -
  • CONR'R by reaction with an appropriate amine in the presence of a suitable coupling agent, such as HATU, in a suitable solvent such as dichloromethane.
  • a suitable coupling agent such as HATU
  • a suitable solvent such as dichloromethane.
  • carboxylic acids may be homologated by one carbon (i.e
  • -OH groups may be generated from the corresponding ester (e.g. -CO 2 R'), or aldehyde (-CHO) by reduction, using for example a complex metal hydride such as lithium aluminium hydride in diethyl ether or tetrahydrofuran, or sodium borohydride in a solvent such as methanol.
  • a complex metal hydride such as lithium aluminium hydride in diethyl ether or tetrahydrofuran, or sodium borohydride in a solvent such as methanol.
  • an alcohol may be prepared by reduction of the corresponding acid (-CO 2 H), using for example lithium aluminium hydride in a solvent such as tetrahydrofuran, or by using borane in a solvent such as tetrahydrofuran.
  • Alcohol groups may be converted into leaving groups, such as halogen atoms or sulfonyloxy groups such as an alkylsulfonyloxy, e.g. trifluoromethylsulfonyloxy or arylsulfonyloxy, e.g. p-toluenesulfonyloxy group using conditions known to those skilled in the art.
  • halogen atoms or sulfonyloxy groups such as an alkylsulfonyloxy, e.g. trifluoromethylsulfonyloxy or arylsulfonyloxy, e.g. p-toluenesulfonyloxy group
  • an alcohol may be reacted with thioyl chloride in a halogenated hydrocarbon (e.g. dichloromethane) to yield the corresponding chloride.
  • a base e.g. triethylamine
  • alcohol, phenol or amide groups may be alkylated by coupling a phenol or amide with an alcohol in a solvent such as tetrahydrofuran in the presence of a phosphine, e.g. triphenylphosphine and an activator such as diethyl-, diisopropyl, or dimethylazodicarboxylate.
  • a phosphine e.g. triphenylphosphine and an activator such as diethyl-, diisopropyl, or dimethylazodicarboxylate.
  • alkylation may be achieved by deprotonation using a suitable base e.g. sodium hydride followed by subsequent addition of an alkylating agent, such as an alkyl halide.
  • Aromatic halogen substituents in the compounds may be subjected to halogen- metal exchange by treatment with a base, for example a lithium base such as n-butyl or t- butyl lithium, optionally at a low temperature, e.g. around -78°C, in a solvent such as tetrahydrofuran, and then quenched with an electrophile to introduce a desired substituent.
  • a base for example a lithium base such as n-butyl or t- butyl lithium
  • a solvent such as tetrahydrofuran
  • an electrophile to introduce a desired substituent.
  • a formyl group may be introduced by using N,N-dimethylformamide as the electrophile.
  • Aromatic halogen substituents may alternatively be subjected to metal (e.g.
  • Aromatic halogen substituents may also undergo nucleophilic displacement following reaction with an appropriate nucleophile such as an amine or an alcohol.
  • such a reaction may be carried out at elevated temperature in the presence of microwave irradiation.
  • the present invention includes a composition (e.g., a pharmaceutical composition) comprising a compound of Formula I (and/or solvates and salts thereof) and a carrier (a pharmaceutically acceptable carrier).
  • a composition e.g., a pharmaceutical composition
  • a carrier e.g., a pharmaceutically acceptable carrier
  • the present invention also includes a composition (e.g., a pharmaceutical composition) comprising a compound of Formula I (and/or solvates and salts thereof) and a carrier (a pharmaceutically acceptable carrier), further comprising a second chemotherapeutic agent and/or a second anti-inflammatory agent such as those described herein.
  • the present compositions are useful for inhibiting abnormal cell growth or treating a hyperproliferative disorder in a mammal (e.g., human).
  • the present compositions are also useful for treating inflammatory diseases in a mammal (e.g., human).
  • the present compounds and compositions are also useful for treating an autoimmune disease, destructive bone disorder, proliferative disorders, infectious disease, viral disease, f ⁇ brotic disease or neurodegenerative disease in a mammal (e.g., human).
  • diseases/disorders include, but are not limited to, diabetes and diabetic complications, diabetic retinopathy, retinopathy of prematurity, age-related macular degeneration, hemangioma, idiopathic pulmonary fibrosis, rhinitis and atopic dermatitis, renal disease and renal failure, polycystic kidney disease, congestive heart failure, neurofibromatosis, organ transplant rejection, cachexia, stroke, septic shock, heart failure, organ transplant rejection, Alzheimer's disease, chronic or neuropathic pain, and viral infections such as HIV, hepatitis (B) virus (HBV), human papilloma virus (HPV), cytomegalovirus (CMV), and Epstein-Barr virus (EBV).
  • HBV hepatitis
  • HPV human papilloma virus
  • CMV cytomegalovirus
  • EBV Epstein-Barr virus
  • Chronic pain for purposes of the present invention includes, but is not limited to, idiopathic pain, and pain associated with chronic alcoholism, vitamin deficiency, uremia, hypothyroidism, inflammation, arthritis, and post-operative pain.
  • Neuropathic pain is associated with numerous conditions which include, but are not limited to, inflammation, postoperative pain, phantom limb pain, burn pain, gout, trigeminal neuralgia, acute herpetic and postherpetic pain, causalgia, diabetic neuropathy, plexus avulsion, neuroma, vasculitis, viral infection, crush injury, constriction injury, tissue injury, limb amputation, arthritis pain, and nerve injury between the peripheral nervous system and the central nervous system.
  • the present compounds and compositions are also useful for treating pancreatitis or kidney disease (including proliferative glomerulonephritis and diabetes- induced renal disease) in a mammal (e.g., human).
  • present compounds and compositions are also useful for the prevention of blastocyte implantation in a mammal (e.g., human).
  • the present invention includes a method of inhibiting abnormal cell growth or treating a hyperproliferative disorder in a mammal (e.g., human) comprising administering to said mammal a therapeutically effective amount of a compound of Formula I (and/or solvates and salts thereof) or a composition thereof. Also included in the present invention is a method of treating an inflammatory disease in a mammal (e.g., human) comprising administering to said mammal a therapeutically effective amount of a compound of Formula I (and/or solvates and/or salts thereof) or a composition thereof.
  • the present invention includes a method of inhibiting abnormal cell growth or treating a hyperproliferative disorder in a mammal (e.g., human) comprising administering to said mammal a therapeutically effective amount of a compound of Formula I (and/or solvates and salts thereof) or a composition thereof, in combination with a second chemotherapeutic agent such as those described herein.
  • the present invention also includes a method of treating an inflammatory disease in a mammal (e.g., human) comprising administering to said mammal a therapeutically effective amount of a compound of Formula I (and/or solvates and/or salts thereof) or a composition thereof, in combination with a second antiinflammatory agent such as those described herein.
  • the present invention includes a method of treating an autoimmune disease, destructive bone disorder, proliferative disorders, infectious disease, viral disease, f ⁇ brotic disease or neurodegenerative disease in a mammal (e.g., human) comprising administering to said mammal a therapeutically effective amount of a compound of Formula I (and/or solvates and salts thereof) or a composition thereof, and optionally further comprising a second therapeutic agent.
  • a mammal e.g., human
  • diseases/disorders include, but are not limited to, diabetes and diabetic complications, diabetic retinopathy, retinopathy of prematurity, age- related macular degeneration, hemangioma, idiopathic pulmonary fibrosis, rhinitis and atopic dermatitis, renal disease and renal failure, polycystic kidney disease, congestive heart failure, neurofibromatosis, organ transplant rejection, cachexia, stroke, septic shock, heart failure, organ transplant rejection, Alzheimer's disease, chronic or neuropathic pain, and viral infections such as HIV, hepatitis (B) virus (HBV), human papilloma virus (HPV), cytomegalovirus (CMV), and Epstein-Barr virus (EBV).
  • HBV hepatitis
  • HPV human papilloma virus
  • CMV cytomegalovirus
  • EBV Epstein-Barr virus
  • the present invention includes a method of treating pancreatitis or kidney disease (including proliferative glomerulonephritis and diabetes-induced renal disease) in a mammal (e.g., human) comprising administering to said mammal a therapeutically effective amount of a compound of Formula I (and/or solvates and salts thereof) or a composition thereof, and optionally further comprising a second therapeutic agent.
  • a mammal e.g., human
  • the present invention includes a method for preventing of blastocyte implantation in a mammal (e.g., human) comprising administering to said mammal a therapeutically effective amount of a compound of Formula I (and/or solvates and salts thereof) or a composition thereof, and optionally further comprising a second therapeutic agent.
  • a mammal e.g., human
  • administering to said mammal a therapeutically effective amount of a compound of Formula I (and/or solvates and salts thereof) or a composition thereof, and optionally further comprising a second therapeutic agent.
  • the present invention includes a method of using the present compounds for in vitro, in situ, and in vivo diagnosis or treatment of mammalian cells, organisms, or associated pathological conditions.
  • this invention further relates to a method for sensitizing abnormal cells in a mammal (e.g., human) to treatment with radiation which comprises administering to said mammal an amount of a compound of Formula I (and/or solvates and salts thereof) or a composition thereof, which amount is effective is sensitizing abnormal cells to treatment with radiation.
  • a mammal e.g., human
  • active compound(s) can be effected by any method that enables delivery of the compounds to the site of action. These methods include oral routes, intraduodenal routes, parenteral injection (including intravenous, subcutaneous, intramuscular, intravascular or infusion), topical, inhalation and rectal administration. [00194] The amount of the active compound administered will be dependent on the subject being treated, the severity of the disorder or condition, the rate of administration, the disposition of the compound and the discretion of the prescribing physician. However, an effective dosage is in the range of about 0.001 to about 100 mg per kg body weight per day, preferably about 1 to about 35 mg/kg/day, in single or divided doses.
  • dosage levels below the lower limit of the aforesaid range may be more than adequate, while in other cases still larger doses may be employed without causing any harmful side effect, provided that such larger doses are first divided into several small doses for administration throughout the day.
  • the active compound may be applied as a sole therapy or in combination with one or more chemotherapeutic agents, for example those described herein. Such conjoint treatment may be achieved by way of the simultaneous, sequential or separate dosing of the individual components of treatment.
  • the pharmaceutical composition may, for example, be in a form suitable for oral administration as a tablet, capsule, pill, powder, sustained release formulations, solution, suspension, for parenteral injection as a sterile solution, suspension or emulsion, for topical administration as an ointment or cream or for rectal administration as a suppository.
  • the pharmaceutical composition may be in unit dosage forms suitable for single administration of precise dosages.
  • the pharmaceutical composition will include a conventional pharmaceutical carrier or excipient and a compound according to the invention as an active ingredient. In addition, it may include other medicinal or pharmaceutical agents, carriers, adjuvants, etc.
  • Exemplary parenteral administration forms include solutions or suspensions of active compounds in sterile aqueous solutions, for example, aqueous propylene glycol or dextrose solutions. Such dosage forms can be suitably buffered, if desired.
  • Suitable pharmaceutical carriers include inert diluents or fillers, water and various organic solvents.
  • the pharmaceutical compositions may, if desired, contain additional ingredients such as flavorings, binders, excipients and the like.
  • tablets containing various excipients, such as citric acid may be employed together with various disintegrants such as starch, alginic acid and certain complex silicates and with binding agents such as sucrose, gelatin and acacia.
  • lubricating agents such as magnesium stearate, sodium lauryl sulfate and talc are often useful for tableting purposes.
  • Solid compositions of a similar type may also be employed in soft and hard filled gelatin capsules. Preferred materials, therefore, include lactose or milk sugar and high molecular weight polyethylene glycols.
  • the active compound therein may be combined with various sweetening or flavoring agents, coloring matters or dyes and, if desired, emulsifying agents or suspending agents, together with diluents such as water, ethanol, propylene glycol, glycerin, or combinations thereof.
  • HM- ⁇ Isolute® HM- ⁇ is a modified form of diatomaceous earth that can efficiently absorb aqueous samples HOBt 1-Hydroxybenzotriazole
  • Si-SPE Pre-packed Isolute® silica flash chromatography cartridge
  • Si-ISCO Pre-packed ISCO® silica flash chromatography cartridge
  • Method A Experiments performed on a Waters Micromass ZQ quadrupole mass spectrometer linked to a Hewlett Packard HPl 100 LC system with diode array detector. This system uses a Higgins Clipeus 5micron Cl 8 100 x 3.0mm column and a 1 ml / minute flow rate.
  • the initial solvent system was 95% water containing 0.1% formic acid (solvent A) and 5% acetonitrile containing 0.1% formic acid (solvent B) for the first minute followed by a gradient up to 5% solvent A and 95% solvent B over the next 14 minutes. The final solvent system was held constant for a further 5 minutes.
  • Method B Experiments performed on a Waters Platform LC quadrupole mass spectrometer linked to a Hewlett Packard HPl 100 LC system with diode array detector and 100 position autosampler using a Phenomenex Luna Cl 8(2) 30 x 4.6mm column and a 2 ml / minute flow rate.
  • the solvent system was 95% water containing 0.1% formic acid (solvent A) and 5% acetonitrile containing 0.1% formic acid (solvent B) for the first 0.50 minutes followed by a gradient up to 5% solvent A and 95% solvent B over the next 4 minutes. The final solvent system was held constant for a further 0.50 minutes.
  • Method C Experiments performed on a PE Sciex API 150 EX quadrupole mass spectrometer linked to a Shimadzu LC-IOAD LC system with diode array detector and 225 position autosampler using a Kromasil C18 50 x 4.6mm column and a 3 ml / minute flow rate.
  • the solvent system was a gradient starting with 100% water with 0.05% TFA (solvent A) and 0% acetonitrile with 0.0375% TFA (solvent B), ramping up to 10% solvent A and 90% solvent B over 4 minutes. The final solvent system was held constant for a further 0.50 minutes.
  • Method D Experiments performed on a Shimadzu LCMS-2010A liquid chromatography mass spectrometer linked to a Shimadzu LC-IOAD VP LC system with diode array detector. Uses a Kromasil 100 5micron Cl 8 50 x 4.6 mm column and a 2.5 ml / minute flow rate.
  • the initial solvent system was 100% water containing 0.05% trifluoroacetic acid (solvent A) and 0% acetonitrile containing 0.05% trifluoroacetic acid (solvent B), followed by a gradient up to 10% solvent A and 90% solvent B over 8 minutes. The final solvent system was held constant for a further 2 minutes.
  • Method E Experiments performed on an Agilent Technologies liquid chromatography mass spectrometer linked to an Agilent Technologies Series 1100 LC system with diode array detector. Uses a Zorbax 3.5micron SB-C18 30 x 2.6 mm column and a 0.5 ml / minute flow rate.
  • the initial solvent system was 95% water containing 0.05% trifluoroacetic acid (solvent A) and 5% acetonitrile containing 0.0375% trifluoroacetic acid (solvent B), followed by a gradient up to 5% solvent A and 95% solvent B over 9 minutes. The final solvent system was held constant for a further 1 minute.
  • IniatiatorTM or OptimizerTM which uses a single-mode resonator and dynamic field tuning, both of which give reproducibility and control. Temperature from 40-250 0 C can be achieved, and pressures of up to 20bar can be reached.
  • Constitutively activated human mutant MEKl expressed in insect cells is used as source of enzymatic activity at a final concentration in the kinase assay of 62.5nM.
  • the assay is carried out for 30 minutes in the presence of 50 ⁇ M ATP using recombinant GST-ERKl produced in E. CoIi as substrate. Phosphorylation of the substrate is detected and quantified using HTRF reagents supplied by Cisbio. These consist of an anti- GST antibody conjugated to allophycocyanin (XL665) and an anti-phospho (Thr202/Tyr204) ERK antibody conjugated to europium-cryptate.
  • the anti-phospho antibody recognises ERKl dually phosphorylated on Thr202 and Tyr204.
  • energy transfer from the cryptate to the allophycocyanin occurs following excitation at 340nm, resulting in fluorescence being emitted that is proportional to the amount of phosphorylated substrate produced. Fluorescence is detected using a multiwell fluorimeter.
  • Compounds are diluted in DMSO prior to addition to assay buffer and the final DMSO concentration in the assay is 1%.
  • the IC50 is defined as the concentration at which a given compound achieves
  • IC50 values are calculated using the XLf ⁇ t software package (version 2.0.5).
  • Constitutively activated human mutant MEKl expressed in insect cells is used as source of enzymatic activity at a final concentration in the kinase assay of 15nM.
  • the assay is carried out for 30 minutes in the presence of 50 ⁇ M ATP using recombinant GST-ERKl produced in E. CoIi as substrate. Phosphorylation of the substrate is detected and quantified using HTRF reagents supplied by Cisbio. These consist of an anti- GST antibody conjugated to allophycocyanin (XL665) and an anti-phospho (Thr202/Tyr204) ERK antibody conjugated to europium-cryptate.
  • the anti-phospho antibody recognises ERKl dually phosphorylated on Thr202 and Tyr204.
  • energy transfer from the cryptate to the allophycocyanin occurs following excitation at 340nm, resulting in fluorescence being emitted that is proportional to the amount of phosphorylated substrate produced. Fluorescence is detected using a multiwell fluorimeter.
  • Compounds are diluted in DMSO prior to addition to assay buffer and the final DMSO concentration in the assay is 1%.
  • the IC50 is defined as the concentration at which a given compound achieves
  • IC50 values are calculated using the XLfit software package (version 2.0.5).
  • the assay is carried out for 30 minutes in the presence of 200 ⁇ M ATP using recombinant GST-MEKl produced in E. CoIi as substrate. Phosphorylation of the substrate is detected and quantified using HTRF, and reagents are supplied by Cisbio. These consist of an anti-GST antibody conjugated to allophycocyanin (XL665) and an anti-phospho (Ser217/Ser221) MEK antibody conjugated to europium-cryptate. The anti-phospho antibody recognises MEK dually phosphorylated on Ser217 and Ser221 or singly phosphorylated on
  • Fluorescence is detected using a multiwell fluorimeter.
  • Compounds are diluted in DMSO prior to addition to assay buffer and the final DMSO concentration in the assay is 1%.
  • the IC 50 is defined as the concentration at which a given compound achieves
  • IC50 values are calculated using the XLfit software package
  • HCT 116 human colorectal carcinoma ATCC
  • A375 human malignant melanoma (ATCC)
  • Both cell lines are maintained in DMEM/F12 (1: 1) media (Gibco) supplemented with 10% FCS at 37 0 C in a 5% CO 2 humidified incubator.
  • Cells are seeded in 96-well plates at 2,000 cells/well and after 24 hours they are exposed to different concentrations of compounds in 0.83% DMSO. Cells are grown for a further 72h, and an equal volume of CellTiter-Glo reagent (Promega) is added to each well.
  • the EC50 is defined as the concentration at which a given compound achieves
  • HCT 116 human colorectal carcinoma ATCC
  • A375 human malignant melanoma (ATCC)
  • Both cell lines are maintained in DMEM/F12 (1: 1) media (Gibco) supplemented with 10% FCS at 37 0 C in a 5% CO 2 humidified incubator.
  • Cells are seeded in 96-well plates at 2,000 cells/well and after 24h they are exposed to different concentrations of compounds in 0.83% DMSO. Cells are grown for a further 2 h or 24h, fixed with formaldehyde (2% final) and permeabilised with methanol.
  • DNA fluorescent dye and detection of cellular p-ERK is performed using an anti-rabbit secondary antibody conjugated to the fluorescent Alexa Fluor 488 dye (Molecular probes).
  • the fluorescence is analysed using the Acumen Explorer (TTP Labtech), a laser-scanning microplate cytometer, and the Alexa Fluor 488 signal is normalised to the PI signal
  • the EC50 is defined as the concentration at which a given compound achieves a signal half way between the baseline and the maximum response. EC50 values are calculated using the XLfit software package (version 2.0.5).
  • Step 4 Ethyl 3-(trifluoromethanesulfonyloxy)-furor3,2-c1pyridine-2- carboxylate
  • Step 5 Ethyl 3-(4-Bromo-2-fluoro-phenylamino)-furor3,2-clpyridine-2- carboxylate
  • reaction mixture was then cooled to room temperature and diluted with ethyl acetate (100 ml).
  • the resultant mixture was then filtered through celite 545 and the celite was washed with an additional 50 ml ethyl acetate.
  • the filtrate was then concentrated and purified by flash chromatography (silica, 120 g column, ISCO, 45 mL/Min, 0-70% Ethyl acetate in hexane in 40 minutes) to afford the title compound as a white solid (2.96 g, 64.5%).
  • Step 1 3-Amino-isonicotinic acid
  • reaction mixture was concentrated and the resultant residue absorbed onto HM-N before being purified by flash chromatography (Si-SPE, pentane: diethyl ether, gradient 80:20 to
  • Step 8 Ethyl 3-(2-fluoro-4-iodo-phenylamino)-furor2.3-c1pyridine-2- carboxylate
  • Step 1 2-Fluoro-4-Trimethylsilanyl-Phenylamine
  • Tetrakis(triphenylphosphine)palladium(0) (1.0 g, 0.9 mmol) was added, the flask fitted with a reflux condenser, and the reaction was heated at reflux for 24-48 hours while a slow stream of nitrogen was passed through a rubber septum placed in the top of the condenser. After cooling to room temperature, the reaction mixture was diluted with ethyl ether (40 mL) and filtered through a plug of silica gel (30 mL of Si ⁇ 2 /ethyl ether slurry packed into a 60 mL fritted glass funnel).
  • the filter cake was washed with ethyl ether (60 mL) and the combined organics were concentrated in vacuo to an orange oil, which was purified by flash chromatography (250 mL silica gel, 98: 1: 1 hexane-CH 2 Cl 2 -ethyl ether), yielding the 2-fluoro-4-trimethylsilylnitrobenzene (5.45 g, 75%) as a yellow-orange oil. [00276] The 2-fluoro-4-trimethylsilylnitrobenzene (5.45g, 25.6 mmol) was then dissolved in ethanol (100 mL), transferred to a Parr shaker bottle, flushed with nitrogen, then charged with 10% Pd-C (0.4 g).
  • the reaction mixture was hydrogenated for Ih on the Parr apparatus (45 psi H 2 ), and then filtered through a plug of Celite. The filter cake was washed with ethanol, and the combined filtrates were concentrated. The resulting residue was purified by flash chromatography (250 mL silica gel, 95:5 hexane-ethyl ether), to afford the title compound as a tan oil (4.31 g, 92%).
  • Step 2 Ethyl 3-(4-Trimethylsilyl-2-fluoro-phenylamino)-furor3,2-clpyridine-
  • the reaction was stirred at -50 0 C under nitrogen for 30 minutes. LC/MS showed that the reaction was complete.
  • the reaction was then quenched at -50 0 C by addition of 200 ml sat. Na 2 S 2 O 3 solution, followed by 100 ml of water.
  • the mixture was then transferred to a sep. funnel and shaken.
  • the mixture was then filtered through filter paper.
  • the black solid on the filter paper was further rinsed with dichloromethane and then discarded.
  • the filtrate was then transferred to a separatory funnel. This was then quickly extracted with dichloromethane (3 X 100 ml).
  • the combined dichloromethane layers were then washed with 170 mL 4M NH 4 OH solution in a sep. funnel.
  • Step 1 Ethyl 3-amino-furor3,2-clpyridine-2-carboxylate
  • Step 2 Ethyl 3-(4-iodo-phenylamino)-furor3,2-c "
  • Step 1 Ethyl 3-(4-Bromo-2-chloro-phenylamino)-furor3,2-clpyridine-2- carboxylate
  • Step 2 Ethyl 3-(2-chloro-4-iodo-phenylamino)-furor3,2-c1pyridine-2- carboxylate
  • reaction mixture was cooled to room temperature, then additional copper(I) iodide (4 mg, 0.02 mmol) and trans-N,N'-dimethyl-l,2-cyclohexane diamine (7 ⁇ l, 0.04 mmol) were added and heating resumed at 115 0 C for 18 hours under an argon atmosphere.
  • the reaction mixture was then cooled to room temperature, diluted with dichloromethane, and washed with a 10% solution of ammonia in water, water then brine.
  • Step 1 Ethyl 3-(4-bromo-2,6-difluoro-phenylamino)-furor3,2-clpyridine-2- carboxylate
  • Step 2 Ethyl 3-(2.6-difluoro-4-iodo-phenylamino)-furo
  • Step 1 Ethyl 3-(4-bromo-2,5-difluoro-phenylamino)-furor3,2-clpyridine-2- carboxylate
  • Step 2 Ethyl 3-(2,5-difluoro-4-iodo-phenylamino)-furor3,2-c "
  • Step 1 Ethyl 7-bromo-3-hvdroxy-furor3,2-c "
  • Step 2 3.7-Dibromo-furo
  • Step 4 3 J-Dibromo-2-(4.4-dimethyl-4.5-dihvdro-oxazol-2-yl)-furor3.2- cipyridine
  • Step 5 r7-Bromo-2-(4.4-dimethyl-4.5-dihvdro-oxazol-2-vn-furor3.2- c1pyridin-3-yl1-(2-fmoro-4-iodo-phenvi)-amine
  • Step 1 Ethyl 4-hydroxy-pyrimidine-5-carboxylate
  • the resultant residue was dissolved in ethyl acetate and washed with water, then brine, before being dried over sodium sulfate and concentrated under reduced pressure to provide a residue.
  • the residue was absorbed on HM-N and purified by flash chromatography (Si-SPE, cyclohexane: ethyl acetate, gradient 100:00 to 40:60) to afford the title compound as a yellow oil (2.67 g, 76%).
  • Step 6 Ethyl 5-(2-fluoro-4-trimethylsilanyl-phenylamino)-furor2,3- dipyrimidine-6-carboxylate
  • Step 7 Ethyl 5-(2-fluoro-4-iodo-phenylamino)-furor2,3-dlpyrimidine-6- carboxylate
  • Step 1 4,5-Dichloropyridine-3-carbaldehvde
  • the reaction was stirred at -7O 0 C for 2 h before the addition of DMF (6.74 ml, 86.8 mmol). The reaction was then stirred at -4O 0 C for 1 h and then allowed to warm to -5 0 C before the careful addition of saturated ammonium chloride solution (50 ml) with rapid stirring over 3 min. The mixture was then partitioned between saturated ammonium chloride (150 ml) and dichloromethane (150 ml) and the layers separated. The aqueous layer was extracted with dichloromethane (2 x 100 ml) and the combined organic layers were dried over magnesium sulfate, then concentrated in vacuo.
  • Step 2 4.5-Dichloropyridine-3-carbaldehyde oxime
  • Step 6 7-Chloro-3-(2-fluoro-4-iodo-phenylamino)-furor3,2-c1pyridine- carboxylic acid ethyl ester
  • Step 1 7-Cvano-3-(2-fluoro-4-trimethylsilanyl-phenylamino)-furor3,2- clpyridine-carboxylic acid ethyl ester
  • Step 2 7-Cyano-3-(2-fluoro-4-iodo-phenylamino)-furo
  • Step 1 (4-Bromo-3-fluoro-benzyloxy)-triisopropyl-silane.
  • Step 2 3-(2-Fluoro-4-triisopropylsilanyloxymethyl-phenylamino)-furo
  • Step 1 l-Bromo-2-fluoro-4-methoxy-benzene.
  • Step 2 3-(2-Fluoro-4-methoxy-phenylamino)-furor3,2-c1pyridine-2- carboxylic acid ethyl ester
  • Step 1 3-(2-Fluoro-4-nitro-phenoxy)-furor3.2-c1pyridine-2-carboxylic acid ethyl ester
  • Step 2 3-(4-Amino-2-fluoro-phenoxy)-furor3,2-c1pyridine-2-carboxylic acid ethyl ester
  • Fe powder (299 mg, 5.36 mmol) was added to a solution of 3-(2-Fluoro-4- nitro-phenoxy)-furo[3,2-c]pyridine-2-carboxylic acid ethyl ester (460 mg, 1.3 mmol) in ethanol (8 mL) and 2N aqueous HCl (8 mL). The reaction mixture was heated to 50 0 C, then cooled to room temperature. The unreacted iron was removed with a magnet, then the reaction mixture was concentrated in vacuo. 10 mL each of water and ethanol were added, followed by solid sodium bicarbonate (1.5g). 3.2 g of silica gel was added and the volatiles were removed in vacuo.
  • Step 3 3-(2-Fluoro-4-iodo-phenoxy)-furor3,2-clpyridine-2-carboxylic acid ethyl ester
  • Step 1 4,5-Dibromonicotinic acid
  • 4,5-Dibromonicotinic acid (32.8 g, 116.7 mmol) was stirred as a suspension in acetonitrile (550 ml) at room temperature and treated portionwise with 1,1' - carbonyldiimidazole (29.87 g, 180 mmol) over 10 minutes. The resulting mixture was stirred for 3 hours at room temperature. After this time ethanol (78 ml) was added and stirring continued for a further 48 hours. The solution was then filtered and the filtrate evaporated in vacuo to give a light brown oil.
  • Step 3 7-Bromo-3-hvdroxyfuror3,2-clpyridine-2-carboxylic acid ethyl ester
  • Trifluoromethanesulfonic anhydride (8.32 ml, 49.66 mmol) in dry DCM (70 ml) was added dropwise to a stirred solution of 7-bromo-3-hydroxyfuro[3,2-c]pyridine-2- carboxylic acid ethyl ester (12.80 g, 44.7 mmol) and pyridine (10.88 ml, 128 mmol) in dry DCM (400 ml) at 5-1O 0 C. The resulting mixture was stirred for 1.5 hours at 5-1O 0 C then allowed to warm to room temperature slowly over 3 hours before being left to stand for 16 hours.
  • Step 5 7-Bromo-3-(2-fluoro-4-trimethylsilanylphenylamino)furor3,2- cipyridine-2-carboxylic acid ethyl ester
  • the mixture was de-gassed again before heating at 115 0 C for 4 hours.
  • the reaction mixture was partitioned between ethyl acetate and water then filtered and the layers separated.
  • the organic layer was washed with water then brine, dried over magnesium sulfate, filtered and evaporated in vacuo to give a brown solid.
  • the solid was purified by flash chromatography (SiO 2 , 30% cyclohexane in DCM ) to give the title compound as a pale yellow solid (7.1 g, 55%).
  • Step 1 3-(2-Fluoro-4-trimethylsilanylphenylamino)-7-methylfuro[3,2- clpyridine-2-carboxylic acid ethyl ester
  • Step 2 3-(2-Fluoro-4-iodophenylamino)-7-methylfuro
  • Step 1 3-Hydroxy-furor3.2-clpyridine-2.7-dicarboxylic acid 2-benzyl ester 7- ethyl ester
  • Step 2 3-Trifluoromethanesulfonyloxy-furor3.2-clpyridine-2.7-dicarboxylic acid 2-benzyl ester 7-ethyl ester
  • Step 3 3-(2-Fluoro-4-trimethylsilanyl-phenylamino)-furor3,2-c1pyridine-2,7- dicarboxylic acid 2-benzyl ester 7-ethyl ester
  • Step 4 3-(2-Fluoro-4-trimethylsilanyl-phenylamino)-furor3,2-c "
  • Step 5 2-((R)-2,2-Dimethyl-rL31dioxolan-4-ylmethoxycarbamoyl)-3-(2- fluoro-4-iodo-phenylamino)-furor3,2-c1pyridine-7-carboxylic acid ethyl ester [00408] To a solution of 2-((R)-2,2-dimethyl-[l,3]dioxolan-4-ylmethoxycarbamoyl)-
  • Step 1 2-Dimethylcarbamoyl-3-hydroxy-furo[3.2-c]pyridine-7-carboxyric acid ethyl ester
  • Step 2 2-Dimethylcarbamoyl-3-trifluoromethanesulfonyloxy-furor3,2- cipyridine-7-carboxylic acid ethyl ester
  • Step 3 2-Dimethylcarbamoyl-3-(2-fluoro-4-trimethylsilanyl-phenylamino)- furo
  • Step 1 4-Chloro-5-fluoro-nicotinic acid ethyl ester
  • Step 4 7-Fluoro-3-(2-fluoro-4-trimethylsilanyl-phenylamino)-furor3,2- clpyridine-2-carboxylic acid ethyl ester
  • Step 5 7-Fluoro-3-(2-fluoro-4-iodo-phenylamino)-furo
  • Step 1 4-Chloro-5-fluoro-pyridine-3-carbaldehyde oxime
  • Step 4 7-Fluoro-3-(2-fluoro-4-trimethylsilanyl-phenylamino)- furo
  • Step 5 7-Fluoro-3-(2-fluoro-4-iodo-phenylamino)-furo
  • Step 1 2,4-Dichloro-nicotinic acid ethyl ester
  • Step 2 4-Chloro-3-hvdroxy-furor3,2-clpyridine-2-carboxylic acid ethyl ester
  • Step 4 4-Chloro-3-(2-fluoro-4-trimethylsilanyl-phenylamino)-furor3,2- cipyridine-2-carboxylic acid ethyl ester
  • Step 5 4-Chloro-3-(2-fluoro-4-iodo-phenylamino)-furor3,2-c1pyridine-2- carboxylic acid ethyl ester
  • Step 1 3-(2-Fluoro-4-trimethylsilanyl-phenylamino)-4-methyl- furor3,21pyridine-2-carboxylic acid ethyl ester
  • Step 2 3-(2-Fluoro-4-iodo-phenylamino)-4-methyl-furor3,2-c1pyridine-2- carboxylic acid ethyl ester
  • tert-Butyl-dimethyl-chlorosilane (0.5 g, 3.21 mmol) was added to a stirred solution of isoxazolidin-4-ol hydrochloride (0.40 g, 3.18 mmol) in DMF (3 mL) and the mixture left to stir at ambient temperature for 2.5 hours. The solvent was evaporated and the residue partitioned between ethyl acetate (50 mL) and water (20 mL). The organic phase was separated, washed with water (3 x 20 mL) then brine (20 mL), dried (MgSO 4 ), filtered and evaporated to provide the desired product as a colourless oil (0.62 g, 96%).
  • Step 1 (S)-3-d.3-Dioxo-1.3-dihvdro-isoindol-2-yloxy)-pyrrolidine-l- carboxylic acid tert-butyl ester
  • Step 2 (S)-3-Aminooxy-pyrrolidine- 1-carboxylic acid tert-butylester
  • Step 1 2-(N-Boc-aminooxy)isobutyric acid ethyl ester
  • Step 1 2-(2-Hvdroxy-2-methyl-propoxy)-isoindole- 1 ,3 -dione
  • Step 1 2-(3-Hvdroxy-3-methyl-butoxy)-isoindole-1.3-dione
  • Step 1 N-Boc-aminooxymethyl(pyridine-2-yl)
  • Step 1 (2-Benzyloxy-l-methyl-ethoxy)-ter?-butyl-dimethyl-silane
  • Step 3 2-
  • N-methylhydrazine (23 ⁇ L, 0.43 mmol) was added to a solution of 2-[2-(tert- Butyl-dimethyl-silanyloxy)-propoxy]-isoindole-l,3-dione (135 mg, 0.40 mmol) in CH 2 CI 2 (3 mL). After stirring for Ih at room temperature, the white precipitate was filtered off and the reaction mixture was concentrated in vacuo to afford the title compound (76 mg, 92% yield) as a yellow oil.
  • Step 1 l-(ter?-Butyl-dimethyl-silanyloxy)-propan-2-ol
  • Step 2 2-r2-(fer ⁇ Butyl-dimethyl-silanyloxy)-l-methyl-ethoxy1-isoindole-l,3- dione
  • Step 1 2-(2-Phenyl-1.3-dioxinan-5-yloxy)-isoindole-1.3-dione
  • Step 1 r2-(l,3-Dioxo-l,3-dihvdro-isoindol-2-yloxy)-ethvH-carbamic acid tert- butyl ester
  • N-hydroxyphthalimide (5.1g, 31.0 mmol) and triphenylphosphine (8.5 g, 32.6 mmol) in tetrahydrofuran (30 ml) at 0 0 C was added dropwise diisopropyl azodicarboxylate (6.3 ml, 32.6 mmol).
  • the reaction was stirred and allowed to warm to room temperature over 16 hours.
  • the reaction was concentrated in vacuo and the product purified by flash chromatography (SiC ⁇ , gradient ethyl acetate:cyclohexane, 20:80 to 30:70) to yield the title compound as an oil (14.2 g).
  • Step 2 2-(2-Amino-ethoxy)-isoindole-l,3-dione
  • Step 3 N-r2- ⁇ .3-Dioxo-1.3-dihvdro-isoindol-2-yloxy)-ethyl1- methanesulfonamide
  • Step 4 N-(2-Aminooxy-ethyl)-methanesulfonamide
  • N-[2-(l,3-dioxo-l,3-dihydro-isoindol-2-yloxy)-ethyl]- ethanesulfonamide (0.55 g, 1.92 mmol) in dichloromethane (15 ml) was added methylhydrazine (0.1 ml, 1.92 mmol). The reaction was stirred at room temperature for 30 minutes, over which time a white precipitate formed. The reaction was filtered and the filtrate concentrated in vacuo to give a residue.
  • Step 1 2-ri-(Toluene-4-sulfonyl)-lH-imidazol-2-ylmethoxy1-isoindole-l,3- dione
  • Step 2 Q-[I -(Toluene-4-sulfonyD- 1 H-imidazol-2-ylmethyl] -hydroxylamine
  • Step 1 GR.4R)-l-benzyl-3.4-dihvdroxypyrrolidine-2.5-dione
  • Step 1 tert-butyl 3-hydroxy-3-methylpyrrolidine-l-carboxylate
  • Step 1 (9H-fluoren-9-vnmethyl GR.4R)-l- ⁇ ert-butoxycarbonvn-4- hvdroxypyrrolidin-3 -ylcarbamate
  • the aqueous layer was extracted once more with ethyl acetate, and the combined organic layers were washed with brine, dried (MgSO 4 ) and concentrated.
  • the residue was purified by silica flash chromatography (gradient elution, using 1 : 1 hexane-ethyl acetate and ethyl acetate) to provide the title compound (0.090 g, 90%).
  • Step 2 (9H-fluoren-9-vnmethyl GR.4R)-4-hvdroxypyrrolidin-3-ylcarbamate hydrochloride
  • Step 1 (2S.3R)-l-(tert-butoxycarbonyl)-3-hydroxypyrrolidine-2-carboxylic acid
  • Step 2 (2R.3R)-tert-butyl 3-hvdroxy-2-(hvdroxymethyl)pyrrolidine-l- carboxylate
  • Step 3 (2R.3R)-2-(hvdroxymethyl)pyrrolidin-3-ol hydrochloride
  • 4N HCl/dioxane solution (5 ml) was added to (2R,3R)-tert-butyl 3-hydroxy-2-
  • Step 1 (2R,4R)-1 -((benzyl oxy)carbonyl)-4-hvdroxypyiroridine-2-carboxyric acid
  • Step 2 (2R, 4R)-I -Benzyl 2 -methyl 4-hvdroxypyrrolidine-L2-dicarboxylate
  • Step 3 (2R.4RVBenzyl 4-hvdroxy-2-(livdroxymethvnpyrrolidine-l- carboxylate
  • 1,2-dicarboxylate (0.41 g, 1.47 mmol) in THF (5 ml) was added LiCl (0.19 g, 4.4 mmol) followed by NaBH 4 (0.17 g, 4.4 mmol).
  • ethanol 10 ml
  • the resulting mixture was stirred at room temperature overnight.
  • the reaction flask was placed in an ice bath, and the cooled milky white solution was acidified to pH 2-3 with 37% HCl.
  • the solution was concentrated, and the residue was partitioned between ethyl acetate and water.
  • the aqueous layer was extracted with ethyl acetate, and the combined organic layers were washed with brine, dried (MgSO 4 ) and concentrated. Purification of the resulting oil by flash chromatography (silica gel, using ethyl acetate) provided the title compound (0.35 g, 95%) as a colorless oil.
  • Step 1 (2R, 4S)-I -benzyl 2 -methyl 4-hvdroxypyrrolidine-l,2-dicarboxylate
  • 1,2-dicarboxylate (1.45 g, 5.2 mmol), triphenylphosphine (4.59 g, 17.5 mmol), and p- nitrobenzoic acid (2.6 g, 15.6 mmol) in dry benzene (10 ml) at room temperature was added diethylazodicarboxylate dropwise (2.57 ml, 17.5 mmol). The solution was then stirred at room temperature for 6 h, whereupon the volatile components were removed in vacuo and the residue purified by flash chromatography (silica gel, hexane-ethyl ether 1: 1, and again with hexane-ethyl ether-methylene chloride 2: 1 : 1).
  • Step 2 (3 S.5R)- 1 -((Benzyloxy)carbonyl)-5-(methoxycarbonyl)pyrrolidin-3 -yl
  • Step 3 (2R,4R)-1 -benzyl 2-methyl 4-azidopyrroridine-l,2-dicarboxylate
  • Triphenylphosphine (0.33 g, 1.15 mmol) was added to a solution of the
  • Step 5 tert-butyl (3R.5R)-l-((benzyloxy)carbonyl)-5-
  • Step 6 tert-butyl GR.5R)-5-(hvdroxymethvnpyrrolidin-3-ylcarbamate
  • Step 1 (2S.3S)-l-(tert-butoxycarbonyl)-3-hydroxypyrrolidine-2-carboxylic acid
  • Step 2 (2S.3 S)- l-tert-Butyl-2 -methyl 3-hydroxypyrrolidine-1.2-dicarboxylate
  • Step 3 (2R.3S)-tert-butyl 3-hvdroxy-2-(hvdroxymethyl)pyrrolidine-l- carboxylate

Abstract

The invention relates to azabenzofuranyl compounds of Formula (I) with anti-cancer and/or anti-inflammatory activity and more specifically to azabenzofuranyl compounds which inhibit MEK kinase activity. The invention provides compositions and methods useful for inhibiting abnormal cell growth or treating a hyperproliferative disorder, or treating an inflammatory disease in a mammal. The invention also relates to methods of using the compounds for in vitro, in situ, and in vivo diagnosis or treatment of mammalian cells, or associated pathological conditions.

Description

AZA-BENZOFURANYL COMPOUNDS AND METHODS OF USE
[0001] RELATED APPLICATIONS
[0002] This application is an international patent application, which claims priority to
U.S. provisional application number 60X839,161 filed August 21, 2006, US provisional application number 60/871,591 filed December 22, 2006, US provisional application number 60/917,623 filed May 11, 2007 and US provisional application number 60/944,741 filed June 18, 2007, the contents of which are incorporated herein by reference. [0003] FIELD OF THE INVENTION
[0004] The invention relates to azabenzofuranyl compounds with anti-cancer and/or anti-inflammatory activity and more specifically to azabenzofuranyl compounds which inhibit MEK kinase activity. The invention also relates to methods of using the compounds for in vitro, in situ, and in vivo diagnosis or treatment of mammalian cells, or associated pathological conditions.
[0005] BACKGROUND OF THE INVENTION
[0006] In the quest to understand how Ras transmits extracellular growth signals, the
MAP (mitogen-activated protein) kinase (MAPK) pathway has emerged as the crucial route between membrane-bound Ras and the nucleus. The MAPK pathway encompasses a cascade of phosphorylation events involving three key kinases, namely Raf, MEK (MAP kinase kinase) and ERK (MAP kinase). Active GTP -bound Ras results in the activation and indirect phosphorylation of Raf kinase. Raf then phosphorylates MEKl and 2 on two serine residues (S218 and S222 for MEKl and S222 and S226 for MEK2) (Ahn et al., Methods in Enzymology 2001, 332, 417-431). Activated MEK then phosphorylates its only known substrates, the MAP kinases, ERKl and 2. ERK phosphorylation by MEK occurs on Y204 and T202 for ERKl and Y 185 and T 183 for ERK2 (Ahn et al., Methods in Enzymology 2001, 332, 417-431). Phosphorylated ERK dimerizes and then translocates to the nucleus where it accumulates (Khokhlatchev et al., Cell 1998, 93, 605-615). In the nucleus, ERK is involved in several important cellular functions, including but not limited to nuclear transport, signal transduction, DNA repair, nucleosome assembly and translocation, and mRNA processing and translation (Ahn et al., Molecular Cell 2000, 6, 1343-1354). Overall, treatment of cells with growth factors leads to the activation of ERKl and 2 which results in proliferation and, in some cases, differentiation (Lewis et al., Adv. Cancer Res. 1998, 74, 49-139). [0007] There has been strong evidence that genetic mutations and/or overexpression of protein kinases involved in the MAP kinase pathway lead to uncontrolled cell proliferation and, eventually, tumor formation, in proliferative diseases. For example, some cancers contain mutations which result in the continuous activation of this pathway due to continuous production of growth factors. Other mutations can lead to defects in the deactivation of the activated GTP -bound Ras complex, again resulting in activation of the MAP kinase pathway. Mutated, oncogenic forms of Ras are found in 50% of colon and >90% pancreatic cancers as well as many others types of cancers (Kohl et al., Science 1993, 260, 1834-1837). Recently, bRaf mutations have been identified in more than 60% of malignant melanoma (Davies, H. et al., Nature 2002, 417, 949-954). These mutations in bRaf result in a constitutively active MAP kinase cascade. Studies of primary tumor samples and cell lines have also shown constitutive or overactivation of the MAP kinase pathway in cancers of pancreas, colon, lung, ovary and kidney (Hoshino, R. et al., Oncogene 1999, 18, 813-822). [0008] MEK has emerged as an attractive therapeutic target in the MAP kinase cascade pathway. MEK, downstream of Ras and Raf, is highly specific for the phosphorylation of MAP kinase; in fact, the only known substrates for MEK phosphorylation are the MAP kinases, ERKl and 2. Inhibition of MEK has been shown to have potential therapeutic benefit in several studies. For example, small molecule MEK inhibitors have been shown to inhibit human tumor growth in nude mouse xenografts, (Sebolt-Leopold et al., Nature-Medicine 1999, 5 (7), 810-816); Trachet et al., AACR Apr. 6-10, 2002, Poster #5426; Tecle, H. IBC 2.sup.nd International Conference of Protein Kinases, Sep. 9-10, 2002), block static allodynia in animals (WO 01/05390 published Jan. 25, 2001) and inhibit growth of acute myeloid leukemia cells (Milella et al., J Clin Invest 2001, 108 (6), 851-859). [0009] Several small molecule MEK inhibitors have also been discussed in, for example, WO02/06213, WO 03/077855 and WO03/077914. There still exists a need for new MEK inhibitors as effective and safe therapeutics for treating a variety of proliferative disease states, such as conditions related to the hyperactivity of MEK, as well as diseases modulated by the MEK cascade.
[0010] SUMMARY OF THE INVENTION
[0011] The invention relates generally to aza-benzofuran compounds of Formula I
(and/or solvates and salts thereof) with anti-cancer and/or anti-inflammatory activity, and more specifically with MEK kinase inhibitory activity. Certain hyperproliferative and inflammatory disorders are characterized by the modulation of MEK kinase function, for example by mutations or overexpression of the proteins. Accordingly, the compounds of the invention and compositions thereof are useful in the treatment of hyperproliferative disorders such as cancer and/or inflammatory diseases such as rheumatoid arthritis.
Figure imgf000004_0001
wherein:
Z1 Is CR1 Or N;
Z2 is CR2 or N;
Z3 is CR3 or N;
Z4 is CR4 or N; where one or two of Z1, Z2, Z3, and Z4 are N;
R1, R2, R3 and R4 are independently selected from H, halo, CN, CF3, -OCF3, -NO2, -(CR14R15)nC(=Y)R11, -(CR14R15)nC(=Y)OR11, -(CR14R15)nC(=Y)NR11R12, -(CR14R15)nNRnR12, -(CR14R15)nORn, -(CR14R1 ^nSR1 \ -(CR14R15)nNR12C(=Y)Rn, -(CR14R15)nNR12C(=Y)ORn, -(CR14R15)nNR13C(=Y)NRnR12, -(CR14R15)nNR12SO2Rn, -(CR14R15)nOC(=Y)R11, -(CR14R15)nOC(=Y)OR11, -(CR14R15)nOC(=Y)NR11R12, -(CR14R15)nOS(O)2(ORn), -(CR14R15)nOP(=Y)(ORn)(OR12), -(CR14R15)nOP(ORn)(OR12), -(CR14R15)nS(O)Rn, -(CR14R15)nS(O)2Rn, -(CR14R15)n S(O)2NR11R12, -(CR14R15)nS(O)(ORn), -(CR14R15)nS(O)2(ORn), -(CR14R15)n SC(=Y)Rn, -(CR14R15)nSC(=Y)ORn, -(CR14R15)nSC(=Y)NRnR12, CJ-CI2 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl;
Figure imgf000004_0002
R5 and R6 are independently selected from H or Ci-Ci2 alkyl;
X1 is selected from R11, -OR11, -NR11R12, -S(O)R11, and -S(O)2R11; when X1 is R11 or -OR11, R11 or -OR11 of X1 and -R5 are optionally taken together with the nitrogen atom to which they are attached to form a 4-7 membered saturated or unsaturated ring having 0-2 additional heteroatoms selected from O, S and N, wherein said ring is optionally substituted with one or more groups selected from halo, CN, CF3, -OCF3, -NO2, oxo, -Si(Ci-Co alkyl), -(CR19R20)nC(=Y')R16, -(CR19R20)n C(=Y')OR16, -(CR19R20)nC(=Y')NR16R17, -(CR19R20)nNR16R17, -(CR19R20)nOR16, -(CR19R20)n-SR16, -(CR19R20)n NR16C(=Y')R17, -(CR19R20)n NR16C(=Y')OR17, -(CR19R20)n NR18C(=Y')NR16R17, -(CR19R20)nNR17SO2R16, -(CR19R20)nOC(=Y')R16, -(CR19R20)nOC(=Y')OR16, -(CR19R20)nOC(=Y')NR16R17, -(CR19R20)nOS(O)2(OR16), -(CR19R20)nOP(=Y')(OR16)(OR17), -(CR19R20)nOP(OR16)(OR17), -(CR19R20)nS(O)R16, -(CR19R20)nS(O)2R16, -(CR19R20)nS(O)2NR16R17, -(CR19R20)nS(O)(OR16), -(CR19R20)n S(O)2(OR16), -(CR19R20)n SC(=Y')R16, -(CR19R20)n SC(=Y')0R16, -(CR19R20X SC(=Y')NR16R17, and R21;
X2 is selected from carbocyclyl, heterocyclyl, aryl, and heteroaryl;
R11, R12 and R13 are independently H, Ci-Ci2 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl, or R11 and R12 together with the nitrogen to which they are attached form a 3-8 membered saturated, unsaturated or aromatic ring having 0-2 heteroatoms selected from O, S and N, wherein said ring is optionally substituted with one or more groups selected from halo, CN, CF3, -OCF3, -NO2, Ci-C6 alkyl, -OH, -SH, -0(Ci-C6 alkyl), -S(Ci-C6 alkyl), -NH2, -NH(Ci-C6 alkyl), -N(Ci-C6 alkyl)2, -SO2(Ci-C6 alkyl), -CO2H, -CO2(Ci-C6 alkyl), -C(O)NH2, -C(O)NH(Ci-C6 alkyl), -C(O)N(Ci-C6 alkyl)2, -N(Ci-C6 alkyl)C(O)(Ci-C6 alkyl), -NHC(0)(CrC6 alkyl), -NHSO2(Ci-C6 alkyl), -N(Ci-C6 alkyl)SO2(Ci-C6 alkyl), -SO2NH2, -SO2NH(Ci-C6 alkyl), -SO2N(Ci-C6 alkyl)2, -OC(O)NH2, -OC(O)NH(Ci-C6 alkyl), -OC(O)N(Ci-C6 alkyl)2, -OC(O)O(Ci-C6 alkyl), -NHC(O)NH(Ci-C6 alkyl), -NHC(O)N(Ci-C6 alkyl)2, -N(Ci-C6 alkyl)C(O)NH(CrC6 alkyl), -N(Ci-C6 alkyl)C(O)N(Cr C6 alkyl)2, -NHC(O)NH(Ci-C6 alkyl), -NHC(O)N(Ci-C6 alkyl)2, -NHC(O)O(Ci-C6 alkyl), and -N(Ci-C6 alkyl)C(O)O(Ci-C6 alkyl);
R14 and R15 are independently selected from H, Ci-Ci2 alkyl, aryl, carbocyclyl, heterocyclyl, and heteroaryl; m and n are independently selected from O, 1, 2, 3, 4, 5, or 6;
Y is independently O, NR11, or S; wherein each said alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl and heteroaryl of R1, R2, R3, R4, R5, R6, X1, X2, R11, R12, R13, R14, and R15 is independently optionally substituted with one or more groups independently selected from halo, CN, CF3, -OCF3, -NO2, oxo, -Si(Ci-C6 alkyl), -(CR19R20)nC(=Y')R16, -(CR19R20)n C(=Y')0R16, -(CR19R20)nC(=Y')NR16R17, -(CR19R20)nNR16R17, -(CR19R20)nOR16, -(CR19R20)n-SR16, -(CR19R20)n NR16C(=Y')R17, -(CR19R20)n NR16C(=Y')OR17, -(CR19R20)n NR18C(=Y')NR16R17, -(CR19R20XNR17SO2R16, -(CR19R20)nOC(=Y')R16, -(CR19R20)nOC(=Y')OR16, -(CR19R20)nOC(=Y')NR16R17, -(CR19R20)nOS(O)2(OR16), -(CR19R20)nOP(=Y')(OR16)(OR17), -(CR19R20)nOP(OR16)(OR17), -(CR19R20)nS(O)R16, -(CR19R20)nS(O)2R16, -(CR19R20XS(O)2NR16R17, -(CR19R20)nS(O)(OR16), -(CR19R20)n S(O)2(OR16), -(CR19R20X SC(=Y')R16, -(CR19R20X SC(=Y')0R16, -(CR19R20X SC(=Y')NR16R17, and R21; each R16, R17 and R18 is independently H, Ci-Ci2 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl, wherein said alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, or heteroaryl is optionally substituted with one or more groups selected from halo, oxo, CN, -OCF3, CF3, -NO2, Ci-C6 alkyl, -OH, -SH, -0(Ci-C6 alkyl), -S(Ci-C6 alkyl), -NH2, -NH(Ci-C6 alkyl), -N(Ci-C6 alkyl)2, -SO2(Ci-C6 alkyl), -CO2H, -CO2(Ci-C6 alkyl), -C(O)NH2, -C(O)NH(Ci-C6 alkyl), -C(O)N(Ci-C6 alkyl)2, -N(Ci-C6 alkyl)C(O)(Ci-C6 alkyl), -NHC(O)(Ci-C6 alkyl), -NHSO2(Ci-C6 alkyl), -N(Ci-C6 alkyl)SO2(Ci-C6 alkyl), -SO2NH2, -SO2NH(Ci-C6 alkyl), -SO2N(Ci-C6 alkyl)2, -OC(O)NH2, -OC(O)NH(Ci-C6 alkyl), -OC(O)N(Ci-C6 alkyl)2, -OC(O)O(Ci-C6 alkyl), -NHC(O)NH(Ci-C6 alkyl), -NHC(O)N(Ci-C6 alkyl)2, -N(Ci-C6 alkyl)C(O)NH(Ci-C6 alkyl), -N(Ci-C6 alkyl)C(O)N(Ci-C6 alkyl)2, -NHC(O)NH(Ci-C6 alkyl), -NHC(O)N(Ci-C6 alkyl)2, -NHC(O)O(Ci-C6 alkyl), and -N(Ci-C6 alkyl)C(O)O(Ci-C6 alkyl); or R16 and R17 together with the nitrogen to which they are attached form a 3-8 membered saturated, unsaturated or aromatic ring having 0-2 heteroatoms selected from O, S and N, wherein said ring is optionally substituted with one or more groups selected from halo, CN, -OCF3, CF3, -NO2, Ci-C6 alkyl, -OH, -SH, -0(Ci-C6 alkyl), -S(Ci-C6 alkyl), -NH2, -NH(Ci-C6 alkyl), -N(Ci-C6 alkyl)2, -SO2(Ci-C6 alkyl), -CO2H, -CO2(Ci-C6 alkyl), -C(O)NH2, -C(O)NH(Ci-C6 alkyl), -C(O)N(Ci-C6 alkyl)2, -N(Ci-C6 alkyl)C(O)(Ci-C6 alkyl), -NHC(0)(CrC6 alkyl), -NHSO2(Ci-C6 alkyl), -N(Ci-C6 alkyl)SO2(Ci-C6 alkyl), -SO2NH2, -SO2NH(Ci-C6 alkyl), -SO2N(Ci-C6 alkyl)2, -OC(O)NH2, -OC(O)NH(Ci-C6 alkyl), -OC(O)N(Ci-C6 alkyl)2, -OC(O)O(Ci-C6 alkyl), -NHC(O)NH(Ci-C6 alkyl), -NHC(O)N(Ci-C6 alkyl)2, -N(Ci-C6 alkyl)C(O)NH(Ci-C6 alkyl), -N(Ci-C6 alkyl)C(O)N(Ci- C6 alkyl)2, -NHC(0)NH(CrC6 alkyl), -NHC(0)N(CrC6 alkyl)2, -NHC(0)0(CrC6 alkyl), and -N(Ci-C6 alkyl)C(O)O(Ci-C6 alkyl);
R19 and R20 are independently selected from H, Ci-Ci2 alkyl, -(CH2)n-aryl, -(CH2)n- carbocyclyl, -(CH2)n-heterocyclyl, and -(CH2)n-heteroaryl;
R21 is C1-C12 alkyl, C2-Cs alkenyl, C2-Cs alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl, wherein each member of R21 is optionally substituted with one or more groups selected from halo, CN, -OCF3, CF3, -NO2, Ci-C6 alkyl, -OH, -SH, -0(Ci-C6 alkyl), -S(Ci-C6 alkyl), -NH2, -NH(Ci-C6 alkyl), -N(Ci-C6 alkyl)2, -SO2(Ci-C6 alkyl), -CO2H, -CO2(Ci-C6 alkyl), -C(O)NH2, -C(0)NH(CrC6 alkyl), -C(0)N(CrC6 alkyl)2, -N(Ci -C6 alkyl)C(O)(Ci-C6 alkyl), -NHC(O)(Ci-C6 alkyl), -NHSO2(Ci-C6 alkyl), -N(Ci-C6 alkyl)SO2(Ci-C6 alkyl), -SO2NH2, -SO2NH(Ci-C6 alkyl), -SO2N(Ci-C6 alkyl)2, -OC(O)NH2, -OC(O)NH(Ci-C6 alkyl), -0C(0)N(CrC6 alkyl)2, -OC(O)O(CrC6 alkyl), -NHC(O)NH(Ci-C6 alkyl), -NHC(O)N(Ci-C6 alkyl)2, -N(Ci-C6 alkyl)C(O)NH(Ci-C6 alkyl), -N(Ci-C6 alkyl)C(O)N(Ci-C6 alkyl)2, -NHC(O)NH(Ci-C6 alkyl), -NHC(O)N(Ci-C6 alkyl)2, -NHC(O)O(Ci-C6 alkyl), and -N(Ci-C6 alkyl)C(O)O(Ci-C6 alkyl); each Y' is independently O, NR22, or S; and
R22 is H or Ci-Ci2 alkyl.
[0012] The present invention includes a composition (e.g., a pharmaceutical composition) comprising a compound of Formula I (and/or solvates, hydrates and/or salts thereof) and a carrier (a pharmaceutically acceptable carrier). The present invention also includes a composition (e.g., a pharmaceutical composition) comprising a compound of Formula I (and/or solvates, hydrates and/or salts thereof) and a carrier (a pharmaceutically acceptable carrier), further comprising a second chemotherapeutic and/or a second antiinflammatory agent. The present compositions are useful for inhibiting abnormal cell growth or treating a hyperproliferative disorder in a mammal (e.g., human). The present compositions are also useful for treating inflammatory diseases in a mammal (e.g., human). [0013] The present invention includes a method of inhibiting abnormal cell growth or treating a hyperproliferative disorder in a mammal (e.g., human) comprising administering to said mammal a therapeutically effective amount of a compound of Formula I (and/or solvates and salts thereof) or a composition thereof, alone or in combination with a second chemotherapeutic agent.
[0014] The present invention includes a method of treating an inflammatory disease in a mammal (e.g., human) comprising administering to said mammal a therapeutically effective amount of a compound of Formula I (and/or solvates and salts thereof) or a composition thereof, alone or in combination with a second anti-inflammatory agent.
[0015] The present invention includes a method of using the present compounds for in vitro, in situ, and in vivo diagnosis or treatment of mammalian cells, organisms, or associated pathological conditions.
[0016] DETAILED DESCRIPTION OF EXEMPLARY EMBODIMENTS
[0017] Reference will now be made in detail to certain embodiments of the invention, examples of which are illustrated in the accompanying structures and formulas. While the invention will be described in conjunction with the enumerated embodiments, it will be understood that they are not intended to limit the invention to those embodiments. On the contrary, the invention is intended to cover all alternatives, modifications, and equivalents which may be included within the scope of the present invention as defined by the claims. One skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in the practice of the present invention. The present invention is in no way limited to the methods and materials described. In the event that one or more of the incorporated literature, patents, and similar materials differs from or contradicts this application, including but not limited to defined terms, term usage, described techniques, or the like, this application controls. [0018] DEFINITIONS
[0019] The term "alkyl" as used herein refers to a saturated linear or branched-chain monovalent hydrocarbon radical of one to twelve carbon atoms. Examples of alkyl groups include, but are not limited to, methyl (Me, -CH3), ethyl (Et, -CH2CH3), 1 -propyl (n-Pr, n- propyl, -CH2CH2CH3), 2-propyl (i-Pr, i-propyl, -CH(CH3)2), 1 -butyl (n-Bu, n-butyl, - CH2CH2CH2CH3), 2 -methyl- 1 -propyl (i-Bu, i-butyl, -CH2CH(CH3)2), 2-butyl (s-Bu, s-butyl, - CH(CH3)CH2CH3), 2-methyl-2-propyl (t-Bu, t-butyl, -C(CH3)3), 1-pentyl (n-pentyl, - CH2CH2CH2CH2CH3), 2-pentyl (-CH(CH3)CH2CH2CH3), 3-pentyl (-CH(CH2CH3)2), 2- methyl-2-butyl (-C(CH3)2CH2CH3), 3-methyl-2-butyl (-CH(CH3)CH(CH3)2), 3 -methyl- 1- butyl (-CH2CH2CH(CH3)2), 2-methyl-l -butyl (-CH2CH(CH3)CH2CH3), 1-hexyl (- CH2CH2CH2CH2CH2CH3), 2-hexyl (-CH(CH3)CH2CH2CH2CH3), 3-hexyl (- CH(CH2CH3)(CH2CH2CH3)), 2-methyl-2-pentyl (-C(CH3)2CH2CH2CH3), 3-methyl-2-pentyl (-CH(CH3)CH(CH3)CH2CH3), 4-methyl-2-pentyl (-CH(CH3)CH2CH(CH3)2), 3-methyl-3- pentyl (-C(CH3)(CH2CH3)2), 2-methyl-3-pentyl (-CH(CH2CH3)CH(CH3)2), 2,3 -dimethyls- butyl (-C(CH3)2CH(CH3)2), 3,3-dimethyl-2-butyl (-CH(CH3)C(CH3)3, 1-heptyl, 1-octyl, and the like.
[0020] The term "alkenyl" refers to linear or branched-chain monovalent hydrocarbon radical of two to twelve carbon atoms with at least one site of unsaturation, i.e., a carbon- carbon, sp double bond, wherein the alkenyl radical includes radicals having "cis" and "trans" orientations, or alternatively, "E" and "Z" orientations. Examples include, but are not limited to, ethylenyl or vinyl (-CH=CH2), allyl (-CH2CH=CH2), and the like. [0021] The term "alkynyl" refers to a linear or branched monovalent hydrocarbon radical of two to twelve carbon atoms with at least one site of unsaturation, i.e., a carbon- carbon, sp triple bond. Examples include, but are not limited to, ethynyl (-C≡CH), propynyl (propargyl, -CH2C≡CH), and the like.
[0022] The terms "carbocycle", "carbocyclyl", "carbocyclic ring" and "cycloalkyl" refer to a monovalent non-aromatic, saturated or partially unsaturated ring having 3 to 12 carbon atoms as a monocyclic ring or 7 to 12 carbon atoms as a bicyclic ring. Bicyclic carbocycles having 7 to 12 atoms can be arranged, for example, as a bicyclo [4,5], [5,5], [5,6] or [6,6] system, and bicyclic carbocycles having 9 or 10 ring atoms can be arranged as a bicyclo [5,6] or [6,6] system, or as bridged systems such as bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane and bicyclo[3.2.2]nonane. Examples of monocyclic carbocycles include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-l-enyl, 1- cyclopent-2-enyl, l-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-l-enyl, l-cyclohex-2-enyl, 1- cyclohex-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, cyclododecyl, and the like.
[0023] "Aryl" means a monovalent aromatic hydrocarbon radical of 6-18 carbon atoms derived by the removal of one hydrogen atom from a single carbon atom of a parent aromatic ring system. Some aryl groups are represented in the exemplary structures as "Ar". Aryl includes bicyclic radicals comprising an aromatic ring fused to a saturated, partially unsaturated ring, or aromatic carbocyclic or heterocyclic ring. Typical aryl groups include, but are not limited to, radicals derived from benzene (phenyl), substituted benzenes, naphthalene, anthracene, indenyl, indanyl, 1 ,2-dihydronapthalene, 1,2,3,4-tetrahydronapthyl, and the like.
[0024] The terms "heterocycle," "heterocyclyl" and "heterocyclic ring" are used interchangeably herein and refer to a saturated or a partially unsaturated (i.e., having one or more double and/or triple bonds within the ring) carbocyclic radical of 3 to 18 ring atoms in which at least one ring atom is a heteroatom selected from nitrogen, oxygen and sulfur, the remaining ring atoms being C, where one or more ring atoms is optionally substituted independently with one or more substituents described below. A heterocycle may be a monocycle having 3 to 7 ring members (2 to 6 carbon atoms and 1 to 4 heteroatoms selected from N, O, P, and S) or a bicycle having 7 to 10 ring members (4 to 9 carbon atoms and 1 to 6 heteroatoms selected from N, O, P, and S), for example: a bicyclo [4,5], [5,5], [5,6], or [6,6] system. Heterocycles are described in Paquette, Leo A.; "Principles of Modern Heterocyclic Chemistry" (W.A. Benjamin, New York, 1968), particularly Chapters 1, 3, 4, 6, 7, and 9; "The Chemistry of Heterocyclic Compounds, A series of Monographs" (John Wiley & Sons, New York, 1950 to present), in particular Volumes 13, 14, 16, 19, and 28; and J. Am. Chem. Soc. (1960) 82:5566. "Heterocyclyl" also includes radicals where heterocycle radicals are fused with a saturated, partially unsaturated ring, or aromatic carbocyclic or heterocyclic ring. Examples of heterocyclic rings include, but are not limited to, pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino, thioxanyl, piperazinyl, homopiperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl, thiazepinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3- dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinylimidazolinyl, imidazolidinyl, 3-azabicyco[3.1.0]hexanyl, 3- azabicyclo[4.1.0]heptanyl, and azabicyclo[2.2.2]hexanyl. Spiro moieties are also included within the scope of this definition. Examples of a heterocyclic group wherein ring atoms are substituted with oxo (=0) moieties are pyrimidinonyl and 1,1-dioxo-thiomorpholinyl. [0025] The term "heteroaryl" refers to a monovalent aromatic radical of 5- or 6- membered rings, and includes fused ring systems (at least one of which is aromatic) of 5-18 atoms, containing one or more heteroatoms independently selected from nitrogen, oxygen, and sulfur. Examples of heteroaryl groups are pyridinyl (including, for example, 2- hydroxypyridinyl), imidazolyl, imidazopyridinyl, pyrimidinyl (including, for example, 4- hydroxypyrimidinyl), pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, triazolyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, and furopyridinyl.
[0026] The heterocycle or heteroaryl groups may be carbon (carbon-linked) or nitrogen (nitrogen-linked) attached where such is possible. By way of example and not limitation, carbon bonded heterocycles or heteroaryls are bonded at position 2, 3, 4, 5, or 6 of a pyridine, position 3, 4, 5, or 6 of a pyridazine, position 2, 4, 5, or 6 of a pyrimidine, position 2, 3, 5, or 6 of a pyrazine, position 2, 3, 4, or 5 of a furan, tetrahydrofuran, thiofuran, thiophene, pyrrole or tetrahydropyrrole, position 2, 4, or 5 of an oxazole, imidazole or thiazole, position 3, 4, or 5 of an isoxazole, pyrazole, or isothiazole, position 2 or 3 of an aziridine, position 2, 3, or 4 of an azetidine, position 2, 3, 4, 5, 6, 7, or 8 of a quinoline or position 1, 3, 4, 5, 6, 7, or 8 of an isoquinoline.
[0027] By way of example and not limitation, nitrogen bonded heterocycles or heteroaryls are bonded at position 1 of an aziridine, azetidine, pyrrole, pyrrolidine, 2- pyrroline, 3-pyrroline, imidazole, imidazolidine, 2-imidazoline, 3 -imidazoline, pyrazole, pyrazoline, 2-pyrazoline, 3-pyrazoline, piperidine, piperazine, indole, indoline, lH-indazole, position 2 of a isoindole, or isoindoline, position 4 of a morpholine, and position 9 of a carbazole, or β-carboline.
[0028] The term "halo" refers to F, Cl, Br or I. The heteroatoms present in heteroaryl or heterocyclcyl include the oxidized forms such as N+→O~, S(O) and S(O)2. [0029] The terms "treat" and "treatment" refer to both therapeutic treatment and prophylactic or preventative measures, wherein the object is to prevent or slow down (lessen) an undesired physiological change or disorder, such as the development or spread of cancer. For purposes of this invention, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable. "Treatment" can also mean prolonging survival as compared to expected survival if not receiving treatment. Those in need of treatment include those already with the condition or disorder as well as those prone to have the condition or disorder or those in which the condition or disorder is to be prevented.
[0030] The phrase "therapeutically effective amount" means an amount of a compound of the present invention that (i) treats or prevents the particular disease, condition, or disorder, (ii) attenuates, ameliorates, or eliminates one or more symptoms of the particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of the particular disease, condition, or disorder described herein. In the case of cancer, the therapeutically effective amount of the drug may reduce the number of cancer cells; reduce the tumor size; inhibit (i.e., slow to some extent and preferably stop) cancer cell infiltration into peripheral organs; inhibit (i.e., slow to some extent and preferably stop) tumor metastasis; inhibit, to some extent, tumor growth; and/or relieve to some extent one or more of the symptoms associated with the cancer. To the extent the drug may prevent growth and/or kill existing cancer cells, it may be cytostatic and/or cytotoxic. For cancer therapy, efficacy can be measured, for example, by assessing the time to disease progression (TTP) and/or determining the response rate (RR).
[0031] The terms "abnormal cell growth" and "hyperproliferative disorder" are used interchangeably in this application. "Abnormal cell growth", as used herein, unless otherwise indicated, refers to cell growth that is independent of normal regulatory mechanisms (e.g., loss of contact inhibition). This includes, for example, the abnormal growth of: (1) tumor cells (tumors) that proliferate by expressing a mutated tyrosine kinase or overexpression of a receptor tyrosine kinase; (2) benign and malignant cells of other proliferative diseases in which aberrant tyrosine kinase activation occurs; (3) any tumors that proliferate by receptor tyrosine kinases; (4) any tumors that proliferate by aberrant serine/threonine kinase activation; and (5) benign and malignant cells of other proliferative diseases in which aberrant serine/threonine kinase activation occurs.
[0032] The terms "cancer" and "cancerous" refer to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth. A "tumor" comprises one or more cancerous cells. Examples of cancer include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia or lymphoid malignancies. More particular examples of such cancers include squamous cell cancer (e.g., epithelial squamous cell cancer), lung cancer including small- cell lung cancer, non-small cell lung cancer ("NSCLC"), adenocarcinoma of the lung and squamous carcinoma of the lung, cancer of the peritoneum, hepatocellular cancer, gastric or stomach cancer including gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial or uterine carcinoma, salivary gland carcinoma, kidney or renal cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma, anal carcinoma, penile carcinoma, acute leukemia, as well as head/brain and neck cancer.
[0033] A "chemotherapeutic agent" is a chemical compound useful in the treatment of cancer. Examples of chemotherapeutic agents include Erlotinib (TARCEVA®, Genentech/OSI Pharm.), Bortezomib (VELCADE®, Millennium Pharm.), Fulvestrant (FASLODEX®, AstraZeneca), Sutent (SUl 1248, Pfizer), Letrozole (FEMARA®, Novartis), Imatinib mesylate (GLEEVEC®, Novartis), PTK787/ZK 222584 (Novartis), Oxaliplatin (Eloxatin®, Sanofi), 5-FU (5-fluorouracil), Leucovorin, Rapamycin (Sirolimus, RAPAMUNE®, Wyeth), Lapatinib (TYKERB®, GSK572016, Glaxo Smith Kline), Lonafarnib (SCH 66336), Sorafenib (BAY43-9006, Bayer Labs), and Gefitinib (IRESSA®, AstraZeneca), AG1478, AG1571 (SU 5271; Sugen), alkylating agents such as thiotepa and CYTOXAN® cyclosphosphamide; alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide and trimethylomelamine; acetogenins (especially bullatacin and bullatacinone); a camptothecin (including the synthetic analog topotecan); bryostatin; callystatin; CC- 1065 (including its adozelesin, carzelesin and bizelesin synthetic analogs); cryptophycins (particularly cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including the synthetic analogs, KW-2189 and CBl-TMl); eleutherobin; pancratistatin; a sarcodictyin; spongistatin; nitrogen mustards such as chlorambucil, chlornaphazine, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimnustine; antibiotics such as the enediyne antibiotics (e.g., calicheamicin, especially calicheamicin gamma II and calicheamicin omegall (Angew Chem. Intl. Ed. Engl. (1994) 33: 183-186); dynemicin, including dynemicin A; bisphosphonates, such as clodronate; an esperamicin; as well as neocarzinostatin chromophore and related chromoprotein enediyne antibiotic chromophores), aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, carabicin, carminomycin, carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin, 6- diazo-5-oxo-L-norleucine, ADRIAMYCIN® (doxorubicin), morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycins, peplomycin, porfϊromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6- mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; anti-adrenals such as aminoglutethimide, mitotane, trilostane; folic acid replenisher such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfornithine; elliptinium acetate; an epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK® polysaccharide complex (JHS Natural Products, Eugene, OR); razoxane; rhizoxin; sizofiran; spirogermanium; tenuazonic acid; triaziquone; 2,2',2"-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A and anguidine); urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxoids, e.g., TAXOL® (paclitaxel; Bristol-Myers Squibb Oncology, Princeton, N.J.), ABRAXANE™ (Cremophor-free), albumin-engineered nanopartic Ie formulations of paclitaxel (American Pharmaceutical Partners, Schaumberg, Illinois), and TAXOTERE® (doxetaxel; Rhδne-Poulenc Rorer, Antony, France); chloranmbucil; GEMZAR® (gemcitabine); 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; etoposide (VP- 16); ifosfamide; mitoxantrone; vincristine; NAVELBINE® (vinorelbine); novantrone; teniposide; edatrexate; daunomycin; aminopterin; capecitabine (XELODA ®); ibandronate; CPT-I l; topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids such as retinoic acid; and pharmaceutically acceptable salts, acids and derivatives of any of the above.
[0034] Also included in the definition of "chemotherapeutic agent" are: (i) anti- hormonal agents that act to regulate or inhibit hormone action on tumors such as anti- estrogens and selective estrogen receptor modulators (SERMs), including, for example, tamoxifen (including NOLVADEX®; tamoxifen citrate), raloxifene, droloxifene, A- hydroxytamoxifen, trioxifene, keoxifene, LYl 17018, onapristone, and FARESTON® (toremifine citrate); (ii) aromatase inhibitors that inhibit the enzyme aromatase, which regulates estrogen production in the adrenal glands, such as, for example, 4(5)-imidazoles, aminoglutethimide, MEGASE® (megestrol acetate), AROMASIN® (exemestane; Pfizer), formestanie, fadrozole, RTVISOR® (vorozole), FEMARA® (letrozole; Novartis), and ARIMIDEX® (anastrozole; AstraZeneca); (iii) anti-androgens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; as well as troxacitabine (a 1,3-dioxolane nucleoside cytosine analog); (iv) protein kinase inhibitors; (v) lipid kinase inhibitors; (vi) antisense oligonucleotides, particularly those which inhibit expression of genes in signaling pathways implicated in aberrant cell proliferation, such as, for example, PKC-alpha, RaIf and H-Ras; (vii) ribozymes such as VEGF expression inhibitors (e.g., ANGIOZYME®) and HER2 expression inhibitors; (viii) vaccines such as gene therapy vaccines, for example, ALLOVECTIN®, LEUVECTΓN®, and VAXID®; PROLEUKIN® rIL-2; a topoisomerase 1 inhibitor such as LURTOTECAN®; ABARELIX® rmRH; (ix) anti-angiogenic agents such as bevacizumab (AVASTIN®, Genentech); and (x) pharmaceutically acceptable salts, acids and derivatives of any of the above. Other anti-angiogenic agents include MMP -2 (matrix- metalloproteinase 2) inhibitors, MMP-9 (matrix-metalloproteinase 9) inhibitors, COX-II (cyclooxygenase II) inhibitors, and VEGF receptor tyrosine kinase inhibitors. Examples of such useful matrix metalloproteinase inhibitors that can be used in combination with the present compounds/compositions are described in WO 96/33172, WO 96/27583, EP 818442, EP 1004578, WO 98/07697, WO 98/03516, WO 98/34918, WO 98/34915, WO 98/33768, WO 98/30566, EP 606,046, EP 931,788, WO 90/05719, WO 99/52910, WO 99/52889, WO 99/29667, WO 99/07675, EP 945864, U.S. Pat. No. 5,863,949, U.S. Pat. No. 5,861,510, and EP 780,386, all of which are incorporated herein in their entireties by reference. Examples of VEGF receptor tyrosine kinase inhibitors include 4-(4-bromo-2-fluoroanilino)-6-methoxy-7- (l-methylpiperidin-4-ylmethoxy)quinazoline (ZD6474; Example 2 within WO 01/32651), 4- (4-fluoro-2-methylindol-5-yloxy)-6-methoxy-7-(3-pyrrolidin-l-ylpropoxy)- quinazoline (AZD2171; Example 240 within WO 00/47212), vatalanib (PTK787; WO 98/35985) and SUl 1248 (sunitinib; WO 01/60814), and compounds such as those disclosed in PCT Publication Nos. WO 97/22596, WO 97/30035, WO 97/32856, and WO 98/13354). [0035] Other examples of chemotherapeutic agents that can be used in combination with the present compounds include inhibitors of PI3K (phosphoinositide-3 kinase), such as those reported in Yaguchi et al (2006) Jour, of the Nat. Cancer Inst. 98(8):545-556; US 7173029; US 7037915; US 6608056; US 6608053; US 6838457; US 6770641; US 6653320; US 6403588; WO 2006/046031; WO 2006/046035; WO 2006/046040; WO 2007/042806; WO 2007/042810; WO 2004/017950; US 2004/092561; WO 2004/007491; WO 2004/006916; WO 2003/037886; US 2003/149074; WO 2003/035618; WO 2003/034997; US 2003/158212; EP 1417976; US 2004/053946; JP 2001247477; JP 08175990; JP 08176070; US 6703414; and WO 97/15658, all of which are incorporated herein in their entireties by reference. Specific examples of such PI3K inhibitors include SF-1126 (PI3K inhibitor, Semafore Pharmaceuticals), BEZ-235 (PI3K inhibitor, Novartis), XL- 147 (PI3K inhibitor, Exelixis, Inc.).
[0036] The term "inflammatory diseases" as used in this application includes, but not limited to, rheumatoid arthritis, atherosclerosis, congestive hear failure, inflammatory bowel disease (including, but not limited to, Crohn's disease and ulcerative colitis), chronic obstructive pulmonary disease in the lung, fϊbrotic disease in the liver and kidney, Crohn's disease, skin diseases such as psoriasis, eczema and scleroderma, osteoarthritis, multiple sclerosis, asthma, diseases and disorders related to diabetic complications, fϊbrotic organ failure in organs such as lung, liver, kidney, and inflammatory complications of the cardiovascular system such as acute coronary syndrome.
[0037] An "anti-inflammatory agent" is a compound useful in the treatment of inflammation. Examples of anti-inflammatory agents include injectable protein therapeutics such as Enbrel®, Remicade®, Humira® and Kineret®. Other examples of anti-inflammatory agents include non-steroidal anti-inflammatory agents (NSAIDs), such as ibuprofen or aspirin (which reduce swelling and alleviate pain); disease-modifying anti-rheumatic drugs (DMARDs) such as methotrexate; 5-aminosalicylates (sulfasalazine and the sulfa-free agents); corticosteroids; immunomodulators such as 6-mercaptoputine ("6-MP"), azathioprine ("AZA"), cyclosporins, and biological response modifiers such as Remicade.RTM. (infliximab) and Enbrel.RTM. (etanercept); fibroblast growth factors; platelet derived growth factors; enzyme blockers such as Arava.RTM. (leflunomide); and/or a cartilage protecting agent such as hyaluronic acid, glucosamine, and chondroitin.
[0038] The term "prodrug" as used in this application refers to a precursor or derivative form of a compound of the invention that is capable of being enzymatically or hydrolytically activated or converted into the more active parent form. See, e.g., Wilman, "Prodrugs in Cancer Chemotherapy" Biochemical Society Transactions, 14, pp. 375-382, 615th Meeting Belfast (1986) and Stella et al., "Prodrugs: A Chemical Approach to Targeted Drug Delivery," Directed Drug Delivery, Borchardt et al., (ed.), pp. 247-267, Humana Press (1985). The prodrugs of this invention include, but are not limited to, ester-containing prodrugs, phosphate-containing prodrugs, thiophosphate-containing prodrugs, sulfate- containing prodrugs, peptide-containing prodrugs, D-amino acid-modified prodrugs, glycosylated prodrugs, β-lactam-containing prodrugs, optionally substituted phenoxyacetamide-containing prodrugs, optionally substituted phenylacetamide-containing prodrugs, 5-fluorocytosine and other 5-fluorouridine prodrugs which can be converted into the more active cytotoxic free drug. Examples of cytotoxic drugs that can be derivatized into a prodrug form for use in this invention include, but are not limited to, compounds of the invention and chemotherapeutic agents such as described above.
[0039] A "metabolite" is a product produced through metabolism in the body of a specified compound or salt thereof. Metabolites of a compound may be identified using routine techniques known in the art and their activities determined using tests such as those described herein. Such products may result for example from the oxidation, hydroxylation, reduction, hydrolysis, amidation, deamidation, esterifϊcation, deesterifϊcation, enzymatic cleavage, and the like, of the administered compound. Accordingly, the invention includes metabolites of compounds of the invention, including compounds produced by a process comprising contacting a compound of this invention with a mammal for a period of time sufficient to yield a metabolic product thereof.
[0040] A "liposome" is a small vesicle composed of various types of lipids, phospholipids and/or surfactant which is useful for delivery of a drug (such as MEK kinase inhibitors disclosed herein and, optionally, a chemotherapeutic agent) to a mammal. The components of the liposome are commonly arranged in a bilayer formation, similar to the lipid arrangement of biological membranes.
[0041] The term "package insert" is used to refer to instructions customarily included in commercial packages of therapeutic products, that contain information about the indications, usage, dosage, administration, contraindications and/or warnings concerning the use of such therapeutic products.
[0042] The term "chiral" refers to molecules which have the property of non- superimposability of the mirror image partner, while the term "achiral" refers to molecules which are superimposable on their mirror image partner.
[0043] The term "stereoisomer" refers to compounds which have identical chemical constitution and connectivity, but different orientations of their atoms in space that cannot be interconverted by rotation about single bonds.
[0044] "Diastereomer" refers to a stereoisomer with two or more centers of chirality and whose molecules are not mirror images of one another. Diastereomers have different physical properties, e.g. melting points, boiling points, spectral properties, and reactivities.
Mixtures of diastereomers may separate under high resolution analytical procedures such as crystallization, electrophoresis and chromatography.
[0045] "Enantiomers" refer to two stereoisomers of a compound which are non- superimposable mirror images of one another. [0046] Stereochemical definitions and conventions used herein generally follow S. P.
Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994. The compounds of the invention may contain asymmetric or chiral centers, and therefore exist in different stereoisomeric forms. It is intended that all stereoisomeric forms of the compounds of the invention, including but not limited to, diastereomers, enantiomers and atropisomers, as well as mixtures thereof such as racemic mixtures, form part of the present invention. Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane-polarized light. In describing an optically active compound, the prefixes D and L, or R and S, are used to denote the absolute configuration of the molecule about its chiral center(s). The prefixes d and 1 or (+) and (-) are employed to designate the sign of rotation of plane-polarized light by the compound, with (-) or 1 meaning that the compound is levorotatory. A compound prefixed with (+) or d is dextrorotatory. For a given chemical structure, these stereoisomers are identical except that they are mirror images of one another. A specific stereoisomer may also be referred to as an enantiomer, and a mixture of such isomers is often called an enantiomeric mixture. A 50:50 mixture of enantiomers is referred to as a racemic mixture or a racemate, which may occur where there has been no stereoselection or stereospecificity in a chemical reaction or process. The terms "racemic mixture" and "racemate" refer to an equimolar mixture of two enantiomeric species, devoid of optical activity. [0047] The term "tautomer" or "tautomeric form" refers to structural isomers of different energies which are interconvertible via a low energy barrier. For example, proton tautomers (also known as prototropic tautomers) include interconversions via migration of a proton, such as keto-enol and imine-enamine isomerizations. Valence tautomers include interconversions by reorganization of some of the bonding electrons. [0048] The phrase "pharmaceutically acceptable salt" as used herein, refers to pharmaceutically acceptable organic or inorganic salts of a compound of the invention. Exemplary salts include, but are not limited, to sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate "mesylate", ethanesulfonate, benzenesulfonate, />-toluenesulfonate, pamoate (i.e., l,l'-methylene-bis -(2-hydroxy-3 -naphthoate)) salts, alkali metal (e.g., sodium and potassium) salts, alkaline earth metal (e.g., magnesium) salts, and ammonium salts. A pharmaceutically acceptable salt may involve the inclusion of another molecule such as an acetate ion, a succinate ion or other counter ion. The counter ion may be any organic or inorganic moiety that stabilizes the charge on the parent compound. Furthermore, a pharmaceutically acceptable salt may have more than one charged atom in its structure. Instances where multiple charged atoms are part of the pharmaceutically acceptable salt can have multiple counter ions. Hence, a pharmaceutically acceptable salt can have one or more charged atoms and/or one or more counter ion.
[0049] If the compound of the invention is a base, the desired pharmaceutically acceptable salt may be prepared by any suitable method available in the art, for example, treatment of the free base with an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, methanesulfonic acid, phosphoric acid and the like, or with an organic acid, such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, a pyranosidyl acid, such as glucuronic acid or galacturonic acid, an alpha hydroxy acid, such as citric acid or tartaric acid, an amino acid, such as aspartic acid or glutamic acid, an aromatic acid, such as benzoic acid or cinnamic acid, a sulfonic acid, such as p-toluenesulfonic acid or ethanesulfonic acid, or the like.
[0050] If the compound of the invention is an acid, the desired pharmaceutically acceptable salt may be prepared by any suitable method, for example, treatment of the free acid with an inorganic or organic base, such as an amine (primary, secondary or tertiary), an alkali metal hydroxide or alkaline earth metal hydroxide, or the like. Illustrative examples of suitable salts include, but are not limited to, organic salts derived from amino acids, such as glycine and arginine, ammonia, primary, secondary, and tertiary amines, and cyclic amines, such as piperidine, morpholine and piperazine, and inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum and lithium. [0051] The phrase "pharmaceutically acceptable" indicates that the substance or composition must be compatible chemically and/or toxicologically, with the other ingredients comprising a formulation, and/or the mammal being treated therewith. [0052] A "solvate" refers to an association or complex of one or more solvent molecules and a compound of the invention. Examples of solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanolamine. The term "hydrate" refers to the complex where the solvent molecule is water.
[0053] The term "protecting group" refers to a substituent that is commonly employed to block or protect a particular functionality while reacting other functional groups on the compound. For example, an "amino-protecting group" is a substituent attached to an amino group that blocks or protects the amino functionality in the compound. Suitable amino- protecting groups include acetyl, trifluoroacetyl, t-butoxycarbonyl (BOC), benzyloxycarbonyl (CBZ) and 9-fluorenylmethylenoxycarbonyl (Fmoc). Similarly, a "hydroxy-protecting group" refers to a substituent of a hydroxy group that blocks or protects the hydroxy functionality. Suitable protecting groups include acetyl and silyl. A "carboxy-protecting group" refers to a substituent of the carboxy group that blocks or protects the carboxy functionality. Common carboxy-protecting groups include phenylsulfonylethyl, cyanoethyl, 2-(trimethylsilyl)ethyl, 2-(trimethylsilyl)ethoxymethyl, 2-(p-toluenesulfonyl)ethyl, 2-(p- nitrophenylsulfenyl)ethyl, 2-(diphenylphosphino)-ethyl, nitroethyl and the like. For a general description of protecting groups and their use, see T. W. Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, New York, 1991.
[0054] The terms "compound of this invention," and "compounds of the present invention" and "compounds of Formula I, ", unless otherwise indicated, include compounds of Formula I and stereoisomers, geometric isomers, tautomers, solvates, metabolites, salts (e.g., pharmaceutically acceptable salts) and prodrugs thereof.
[0055] The present invention provides azabenzofuranyl compounds of Formula I as described above useful as kinase inhibitors, particularly useful as MEK kinase inhibitors. The present invention includes compounds of Formulae I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, I-i, II-a, II-b, II-c, II-d, II-e, II-f, II-g, II-h, II-i, III-a, III-b, III-c, III-d, III-e, III-f, III-g, III-h, and III-i, and all other variables are as defined in Formula I.
Figure imgf000020_0001
Figure imgf000021_0001
Figure imgf000022_0001
[0056] In an embodiment of the present invention, compounds are of Formulae I-b, I- f, I-g, I-h, II-b, II-f, II-g, II-h, III-b, III-f, III-g and III-h, and all other variables are as defined in Formula I.
[0057] In an embodiment of the present invention, compounds are of Formula III-c and all other variables are as defined in Formula I.
[0058] In an embodiment of the present invention, R1 is H, halo, CN, CF3, -NR11R12,
-OR11, -SR11, -C(=O)NRπR12, or Ci-C6 alkyl, and all other variables are as defined in
Formula I, I-a, I-b, I-d, I-f, I-g, II-a, II-b, II-d, II-f, II-g, III-a, III-b, III-d, III-f, or III-g.
[0059] In another embodiment of the present invention, R1 is H, halo, CN, CF3,
Ci-C6 alkyl, -NR11R12 wherein R11 and R12 are independently H or Ci-C6 alkyl, -OR11 wherein R11 is H or Ci-C6 alkyl, or -SR11 wherein R11 is H or Ci-C6 alkyl; and all other variables are as defined in Formula I, I-a, I-b, I-d, I-f, I-g, II-a, II-b, II-d, II-f, II-g, III-a, III-b,
III-d, III-f, or III-g.
[0060] In another embodiment of the present invention, R1 is H, Cl, CN, CF3, methyl,
-NH2, -NH(CH3), -N(CH3)2, -OH, or -OCH3; and all other variables are as defined in
Formula I, I-a, I-b, I-d, I-f, I-g, II-a, II-b, II-d, II-f, II-g, III-a, III-b, III-d, III-f, or III-g.
[0061] In another embodiment of the present invention, R1 is H; and all other variables are as defined in Formula I, I-a, I-b, I-d, I-f, I-g, II-a, II-b, II-d, II-f, II-g, III-a, III-b,
III-d, III-f, or III-g.
[0062] In an embodiment of the present invention, R2 is H, halo, CN, CF3, -NR11R12,
-OR11, -SR11, -C(=O)NRπR12, or Ci-C6 alkyl, and all other variables are as defined in
Formula I, I-a, I-c, I-d, I-e, I-i, II-a, II-c, II-d, II-e, II-i, III-a, III-c, III-d, III-e, or III-i, or as defined above.
[0063] In another embodiment of the present invention, R is H, halo, CN, CF3,
Ci-C6 alkyl, -NR11R12 wherein R11 and R12 are independently H or Ci-C6 alkyl, -OR11 wherein R11 is H or Ci -C6 alkyl, or -SR11 wherein R11 is H or Ci -C6 alkyl; and all other variables are as defined in Formula I, I-a, I-c, I-d, I-e, I-i, II-a, II-c, II-d, II-e, II-i, III-a, III-c,
III-d, III-e, or III-i, or as defined above.
[0064] In another embodiment of the present invention, R2 is H, Cl, CN, CF3, methyl,
-NH2, -NH(CH3), -N(CH3)2, -OH, or -OCH3; and all other variables are as defined in
Formula I, I-a, I-c, I-d, I-e, I-i, II-a, II-c, II-d, II-e, II-i, III-a, III-c, III-d, III-e, or III-i, or as defined above.
[0065] In an embodiment of the present invention, R3 is H, halo, CN, CF3, -NR11R12,
-OR11, -SR11, -C(=O)NRnR12, or Ci-C6 alkyl, and all other variables are as defined in
Formula I, I-a, I-c, I-d, I-e, I-i, II-a, II-c, II-d, II-e, II-i, III-a, III-c, III-d, III-e, or III-i, or as defined above.
[0066] In another embodiment of the present invention, R3 is H, halo, CF3, Ci-C6 alkyl; and all other variables are as defined in Formula I, I-a, I-c, I-d, I-e, I-i, II-a, II-c, II-d,
II-e, II-i, III-a, III-c, III-d, III-e, or III-i, or as defined above.
[0067] In another embodiment of the present invention, R3 is H, F, CF3, or methyl; and all other variables are as defined in Formula I, I-a, I-c, I-d, I-e, I-i, II-a, II-c, II-d, II-e, II- i, III-a, III-c, III-d, III-e, or III-i, or as defined above.
[0068] In another embodiment of the present invention, R is H, F, Cl, CF3, methyl or
CN; and all other variables are as defined in Formula I, I-a, I-c, I-d, I-e, I-i, II-a, II-c, II-d, II- e, II-i, III-a, III-c, III-d, III-e, or III-i, or as defined above.
[0069] In an embodiment of the present invention, R4 is H, halo, CN, CF3, -NR11R12,
-OR11, -SR11, -C(=O)NRπR12, or Ci-C6 alkyl, and all other variables are as defined in
Formula I, I-a, I-b, I-c, I-e, I-g, I-h, II-a, II-b, II-c, II-e, II-g, II-h, III-a, III-b, III-c, III-e, III-g, or III-h, or as defined above.
[0070] In another embodiment of the present invention, R4 is H, halo, CN, CF3,
-NR11R12 or -CC=O)NR11R12 wherein R11 and R12 are independently H or Ci-C6 alkyl, -OR11 wherein R11 is H or Ci -C6 alkyl, or -SR11 wherein R11 is H or Ci -C6 alkyl; and all other variables are as defined in Formula I, I-a, I-b, I-c, I-e, I-g, I-h, II-a, II-b, II-c, II-e, II-g, II-h,
III-a, III-b, III-c, III-e, III-g, or III-h, or as defined above.
[0071] In another embodiment of the present invention, R4 is H, Br, Cl, CN, CF3,
-NH2, -NH(CH3), -N(CH3)2, -C(O)NH2, -C(O)NHCH3, -C(O)N(CH3)2, -OH, or -OCH3; and all other variables are as defined in Formula I, I-a, I-b, I-c, I-e, I-g, I-h, II-a, II-b, II-c, II- e, II-g, II-h, III-a, III-b, III-c, III-e, III-g, or III-h, or as defined above. [0072] In another embodiment of the present invention, R4 is H, Br, Cl, CN, CF3,
-NH2, -NH(CH3), -N(CH3)2, -C(O)NH2, -C(O)NHCH3, -C(O)N(CH3)2, -OH, or -OCH3; and all other variables are as defined in Formula I, I-a, I-b, I-c, I-e, I-g, I-h, II-a, II-b, II-c, II- e, II-g, II-h, Ill-a, III-b, III-c, III-e, III-g, or III-h, or as defined above.
[0073] In another embodiment of the present invention, R4 is halo, -OH, or Ci-C6 alkyl optionally substituted by halo; and all other variables are as defined in Formula I, I-a, I- b, I-c, I-e, I-g, I-h, II-a, II-b, II-c, II-e, II-g, II-h, III-a, III-b, III-c, III-e, III-g, or III-h, or as defined above.
[0074] In another embodiment of the present invention, R4 is independently Cl, Br,
Me, Et, F, CHF2, CF3, or -OH; and all other variables are as defined in Formula I, I-a, I-b, I-c,
I-e, I-g, I-h, II-a, II-b, II-c, II-e, II-g, II-h, III-a, III-b, III-c, III-e, III-g, or III-h, or as defined above.
[0075] In an embodiment of the present invention, R5 is H or Ci-C6 alkyl; and all other variables are as defined in Formula I, I-a to I-i, or II-a to II-i, or as defined above.
[0076] In another embodiment of the present invention, R5 is H or methyl; and all other variables are as defined in Formula I, I-a to I-i, or II-a to II-i, or as defined above.
[0077] In another embodiment of the present invention, R5 is H; and all other variables are as defined in Formula I, I-a to I-i, or II-a to II-i, or as defined above.
[0078] In another embodiment of the present invention, R5 is methyl; and all other variables are as defined in Formula I, I-a to I-i, or II-a to II-i, or as defined above.
[0079] In an embodiment of the present invention, R6 is H or Ci-C6 alkyl; and all other variables are as defined in Formula I, I-a to I-i, II-a to II-i, or III-a to III-i, or as defined above.
[0080] In another embodiment of the present invention, R6 is H or methyl; and all other variables are as defined in Formula I, I-a to I-i, II-a to II-i, or III-a to III-i, or as defined above.
[0081] In another embodiment of the present invention, R6 is H; and all other variables are as defined in Formula I, I-a to I-i, II-a to II-i, or III-a to III-i, or as defined above.
[0082] In another embodiment of the present invention, R6 is methyl; and all other variables are as defined in Formula I, I-a to I-i, II-a to II-i, or III-a to III-i, or as defined above.
[0083] In an embodiment of the present invention, X1 is OR11 (i.e., Formula II-a to II- i); and all other variables are as defined in Formula I or I-a to I-i; or as defined above. [0084] In an embodiment of the present invention, X1 is OR11 wherein R11 is H; and all other variables are as defined in Formula I or I-a to I-i; or as defined above. [0085] In another embodiment of the present invention, X1 is OR11 wherein R11 is Ci-
C12 alkyl (e.g., C1-Ce alkyl) substituted with one or more groups independently selected from halo, CN, CF3, -OCF3, -NO2, oxo, -Si(Cj-C6 alkyl), -(CR19R20)n C(=Y')R16, -(CR19R20)nC(=Y')OR16, -(CR19R20)nC(=Y')NR16R17, -(CR19R20)nNR16R17, -(CR19R20)nOR16, -(CR19R20)n-SR16, -(CR19R20)n NR16C(=Y')R17, -(CR19R20)n NR16C(=Y')OR17, -(CR19R20)n NR18C(=Y')NR16R17, -(CR19R20)nNR17SO2R16, -(CR19R20)nOC(=Y')R16, -(CR19R20)nOC(=Y')OR16, -(CR19R20)n OC(=Y')NR16R17, -(CR19R20)n OS(O)2(OR16), -(CR19R20)nOP(=Y')(OR16)(OR17), -(CR19R20)nOP(OR16)(OR17), -(CR19R20)nS(O)R16, -(CR19R20)n S(O)2R16, -(CR19R20)n S(O)2NR16R17, -(CR19R20)n S(O)(OR16), -(CR19R20)n S(O)2(OR16), -(CR19R20)n SC(=Y')R16, -(CR19R20)n SC(=Y')OR16, -(CR19R20)n SC(=Y')NR16R17, and R21; and all other variables are as defined in Formula I or I-a to I-i, or as defined above.
[0086] In another embodiment of the present invention, X1 is:
Figure imgf000025_0001
Figure imgf000026_0001
other variables are as defined in Formula I or I-a to I-i, or as defined above. [0087] In another embodiment of the present invention, X1 is
Figure imgf000026_0002
; and all other variables are as defined in Formula I or I-a to I-i, or as defined above. [0088] In another embodiment of the present invention, X1 is
Figure imgf000027_0001
all other variables are as defined in Formula I or I-a to I-i, or as defined above.
Figure imgf000027_0002
; and all other variables are as defined in Formula I or I-a to I-i, or as defined above. [0090] In another embodiment of the present invention, X1 is
Figure imgf000028_0001
all other variables are as defined in Formula I or I-a to I-i, or as defined above. [0091] In another embodiment of the present invention, X1 is OR11 wherein R11 is heterocyclyl (e.g., 4- to 6-membered heterocyclyl) optionally substituted with one or more groups independently selected from halo, CN, CF3, -OCF3, -NO2, oxo, -Si(Ci-Co alkyl),
-(CR19R20)nC(=Y')R16, -(CR19R20)nC(=Y')OR16, -(CR19R20)nC(=Y')NR16R17, -(CR19R20)nNR16R17, -(CR19R20)nOR16, -(CR19R20)n-SR16, -(CR19R20)nNR16C(=Y')R17, -(CR19R20)n NR16C(=Y')OR17, -(CR19R20)nNR18C(=Y')NR16R17, -(CR19R20^NR17SO2R16, -(CR19R20)nOC(=Y')R16, -(CR19R20)nOC(=Y')OR16, -(CR19R20)nOC(=Y')NR16R17, -(CR19R20)nOS(O)2(OR16), -(CR19R20)nOP(=Y')(OR16)(OR17), -(CR19R20)nOP(OR16)(OR17), -(CR19R20)nS(O)R16, -(CR19R20)nS(O)2R16, -(CR19R20)nS(O)2NR16R17, -(CR19R20XS(O)(OR16), -(CR19R20X S(O)2(OR16), -(CR19R20)nSC(=Y')R16,
-(CRiyRzυ)nSC(=Y')0R , -(CRiyRzuX SC(=Y')NRlbR1 , and RZi; and all other variables are as defined in Formula I or I-a to I-i, or as defined above.
[0092] In another embodiment of the present invention, X1 is OR11 wherein R11 is 4- to 6-membered heterocyclyl having 1 nitrogen ring atom wherein said heterocyclyl is optionally substituted with one or more groups independently selected from halo, CN, CF3, -OCF3, -NO2, oxo, -Si(Ci-C6 alkyl), -(CR19R20)nC(=Y')R16, -(CR19R20)n C(=Y')OR16, -(CR19R20)nC(=Y')NR16R17, -(CR19R20)nNR16R17, -(CR19R20)nOR16, -(CR19R20)n-SR16, -(CR19R20)nNR16C(=Y')R17, -(CR19R20)nNR16C(=Y')OR17,
-(CR19R20)nNR18C(=Y')NR16R17, -(CR19R20)nNR17SO2R16, -(CR19R20)nOC(=Y')R16, -(CR19R20)nOC(=Y')OR16, -(CR19R20)nOC(=Y')NR16R17, -(CR19R20)nOS(O)2(OR16), -(CR19R20)nOP(=Y')(OR16)(OR17), -(CR19R20)nOP(OR16)(OR17), -(CR19R20XS(O)R16, -(CR19R20XS(O)2R16, -(CR19R20XS(O)2NR16R17, -(CR19R20)nS(O)(OR16), -(CR19R20X S(O)2(OR16), -(CR19R20)nSC(=Y')R16, -(CR19R20)nSC(=Y')OR16, -(CR19R20X SC(=Y')NR16R17, and R21; and all other variables are as defined in Formula I or I-a to I-i, or as defined above.
[0093] In another embodiment of the present invention, X1 is
Figure imgf000029_0001
.
or
Figure imgf000029_0002
; and all other variables are as defined in Formula I or I-a to I-i, or as defined above.
[0094] In another embodiment of the present invention, X1 is
Figure imgf000029_0003
all other variables are as defined in Formula I or I-a to I-i, or as defined above. [0095] In another embodiment of the present invention, X1 is
Figure imgf000029_0004
and all other variables are as defined in
Formula I or I-a to I-i, or as defined above. [0096] In an embodiment of the present invention, X1 is R11, and X1 and R5 are taken together with the nitrogen atom to which they are attached to form a 5-7 membered saturated or unsaturated cyclic ring having 0-2 additional heteroatoms selected from O, S and N, wherein said cyclic ring is optionally substituted with one or more groups selected from halo, CN, CF3, -OCF3, -NO2, oxo, -Si(Cj-C6 alkyl), -(CR19R20)nC(=Y')R16, -(CR19R20)n C(=Y')0R16, -(CR19R20)nC(=Y')NR16R17, -(CR19R20)nNR16R17, -(CR19R20)nOR16, -(CR19R20)n-SR16, -(CR19R20)n NR16C(=Y')R17, -(CR19R20)n NR16C(=Y')OR17, -(CR19R20)n NR18C(=Y')NR16R17, -(CR19R20)nNR17SO2R16, -(CR19R20)nOC(=Y')R16, -(CR19R20)nOC(=Y')OR16, -(CR19R20)nOC(=Y')NR16R17, -(CR19R20)nOS(O)2(OR16), -(CR19R20)nOP(=Y')(OR16)(OR17), -(CR19R20)nOP(OR16)(OR17), -(CR19R20)nS(O)R16, -(CR19R20)nS(O)2R16, -(CR19R20)nS(O)2NR16R17, -(CR19R20)nS(O)(OR16), -(CR19R20)n S(O)2(OR16), -(CR19R20)n SC(=Y')R16, -(CR19R2V SC(=Y')OR16, -(CR19R20)n SC(=Y')NR16R17, and R21; and all other variables are as defined in Formula I or I-a to I-i, or as defined above.
[0097] In another embodiment of the present invention, X1 is R11, and X1 and R5 are taken together with the nitrogen atom to which they are attached to form a 5-6 membered saturated cyclic ring having 0-2 additional heteroatoms selected from O, S and N, wherein said cyclic ring is optionally substituted with one or more groups selected from halo, CN, CF3, -OCF3, -NO2, oxo, -Si(Ci-C6 alkyl), -(CR19R20)nC(=Y')R16, -(CR19R20)n C(=Y')0R16, -(CR19R20)nC(=Y')NR16R17, -(CR19R20)nNR16R17, -(CR19R20)nOR16, -(CR19R20)n-SR16, -(CR19R20X NR16C(=Y')R17, -(CR19R20X NR16C(=Y')OR17, -(CR19R20X NR18C(=Y')NR16R17, -(CR19R20XNR17SO2R16, -(CR19R20)nOC(=Y')R16, -(CR19R20)nOC(=Y')OR16, -(CR19R20χθC(=Y')NR16R17, -(CR19R20XOS(O)2(OR16), -(CR19R20)nOP(=Y')(OR16)(OR17), -(CR19R20χθP(OR16)(OR17), -(CR19R20XS(O)R16, -(CR19R20XS(O)2R16, -(CR19R20XS(O)2NR16R17, -(CR19R20)nS(O)(OR16), -(CR19R20X S(O)2(OR16), -(CR19R20X SC(=Y')R16, -(CR19R20X SC(=Y')OR16, -(CR19R20X SC(=Y')NR16R17, and R21; and all other variables are as defined in Formula I or I-a to I-i, or as defined above. [0098] In another embodiment of the present invention, W is:
Figure imgf000031_0001
all other variables are as defined in Formula I or I-a to I-i, or as defined above. [0099] In another embodiment of the present invention, W is:
Figure imgf000031_0002
other variables are as defined in Formula I or I-a to I-i, or as defined above. [00100] In another embodiment of the present invention, W is:
Figure imgf000031_0003
; and all other variables are as defined in Formula I or I-a to I-i, or as defined above. [00101] In an embodiment of the present invention, X1 is R11, and X1 and R5 are taken together with the nitrogen atom to which they are attached to form a 4-membered saturated or unsaturated cyclic ring having 0-1 additional heteroatoms selected from O, S and N, wherein said cyclic ring is optionally substituted with one or more groups selected from halo, CN, CF3, -OCF3, -NO2, oxo, -Si(Ci-C6 alkyl), -(CR19R20)nC(=Y')R16, -(CR19R20)n C(=Y')0R16, -(CR19R20)nC(=Y')NR16R17, -(CR19R20)nNR16R17, -(CR19R20)nOR16, -(CR19R20)n-SR16, -(CR19R20)n NR16C(=Y')R17, -(CR19R20X NR16C(=Y')OR17, -(CR19R20)n
NR18C(=Y')NR16R17, -(CR19R20XNR17SO2R16, -(CR19R20)nOC(=Y')R16, -(CR19R20)nOC(=Y')OR16, -(CR19R20)nOC(=Y')NR16R17, -(CR19R20XOS(O)2(OR16), -(CR19R20)nOP(=Y')(OR16)(OR17), -(CR19R20χθP(OR16)(OR17), -(CR19R20XS(O)R16,
-(CR19R20XS(O)2R16, -(CR19R20XS(O)2NR16R17, -(CR19R20)nS(O)(OR16), -(CR19R20X S(O)2(OR16), -(CR19R20)n SC(=Y')R16, -(CR19R20)n SC(=Y')OR16, -(CR19R20)n SC(=Y')NR16R17, and R21; and all other variables are as defined in Formula I or I-a to I-i, or as defined above. [00102] In another embodiment of the present invention, W is:
Figure imgf000032_0001
HO ; and all other variables are as defined in Formula I or I-a to I-i, or as above.
[00103] In an embodiment of the present invention, X1 is -OR11, and -OR11 of X1 and
R5 are taken together with the nitrogen atom to which they are attached to form a 4-7 membered saturated or unsaturated cyclic ring having 0-2 additional heteroatoms selected from O, S and N, wherein said cyclic ring is optionally substituted with one or more groups selected from halo, CN, CF3, -OCF3, -NO2, oxo, -Si(Cj-C6 alkyl), -(CR19R20)nC(=Y')R16, -(CR19R20)n C(=Y')OR16, -(CR19R20)nC(=Y')NR16R17, -(CR19R20)nNR16R17, -(CR19R20)nOR16, -(CR19R20)n-SR16, -(CR19R20)n NR16C(=Y')R17, -(CR19R20)n NR16C(=Y')OR17, -(CR19R20)n NR18C(=Y')NR16R17, -(CR19R20)nNR17SO2R16, -(CR19R20)nOC(=Y')R16, -(CR19R20)nOC(=Y')OR16, -(CR19R20)nOC(=Y')NR16R17, -(CR19R20)nOS(O)2(OR16), -(CR19R20)nOP(=Y')(OR16)(OR17), -(CR19R20)nOP(OR16)(OR17), -(CR19R20)nS(O)R16, -(CR19R20)nS(O)2R16, -(CR19R20)nS(O)2NR16R17, -(CR19R20)nS(O)(OR16), -(CR19R20)n S(O)2(OR16), -(CR19R20)n SC(=Y')R16, -(CR19R20)n SC(=Y')OR16, -(CR19R20X SC(=Y')NR16R17, and R21; and all other variables are as defined in Formula I or I-a to I-i, or as defined above.
[00104] In another embodiment of the present invention, X1 is -OR11, and -OR11 of X1 and R5 are taken together with the nitrogen atom to which they are attached to form a 5-7 membered saturated or unsaturated cyclic ring having 0-2 additional heteroatoms selected from O, S and N, wherein said cyclic ring is optionally substituted with one or more groups selected from halo, CN, CF3, -OCF3, -NO2, oxo, -Si(Ci-C6 alkyl), -(CR19R20)nC(=Y')R16, -(CR19R20X C(=Y')OR16, -(CR19R20)nC(=Y')NR16R17, -(CR19R20)nNR16R17, -(CR19R20XOR16, -(CR19R20X-SR16, -(CR19R20X NR16C(=Y')R17, -(CR19R20X NR16C(=Y')OR17, -(CR19R20X NR18C(=Y')NR16R17, -(CR19R20XNR17SO2R16, -(CR19R20)nOC(=Y')R16, -(CR19R20XOC(=Y')OR16, -(CR19R20)nOC(=Y')NR16R17, -(CR19R20XOS(O)2(OR16), -(CR19R20)nOP(=Y')(OR16)(OR17), -(CR19R20χθP(OR16)(OR17),
-(CR19R20XS(O)R16, -(CR19R20XS(O)2R16, -(CR19R20XS(O)2NR16R17, -(CR19R20)nS(O)(OR16), -(CR19R20)n S(O)2(OR16), -(CR19R20)n SC(=Y')R16, -(CR19R20)n SC(=Y')OR16, -(CR19R20X SC(=Y')NR16R17, and R21; and all other variables are as defined in Formula I or I-a to I-i, or as defined above
[00105] In another embodiment of the present invention, X1 is -OR11, and -OR11 of X1 and R5 are taken together with the nitrogen atom to which they are attached to form a 5-6 membered saturated cyclic ring having 0-2 additional heteroatoms selected from O, S and N, wherein said cyclic ring is optionally substituted with one or more groups selected from halo, CN, CF3, -OCF3, -NO2, oxo, -Si(Cj-C6 alkyl), -(CR19R20)nC(=Y')R16, -(CR19R20)n C(=Y')0R16, -(CR19R20)nC(=Y')NR16R17, -(CR19R20)nNR16R17, -(CR19R20)nOR16, -(CR19R20)n-SR16, -(CR19R20)n NR16C(=Y')R17, -(CR19R20)n NR16C(=Y')OR17, -(CR19R20)n NR18C(=Y')NR16R17, -(CR19R20)nNR17SO2R16, -(CR19R20)nOC(=Y')R16, -(CR19R20)nOC(=Y')OR16, -(CR19R20)nOC(=Y')NR16R17, -(CR19R20)nOS(O)2(OR16), -(CR19R20)nOP(=Y')(OR16)(OR17), -(CR19R20)nOP(OR16)(OR17), -(CR19R20XS(O)R16, -(CR19R20XS(O)2R16, -(CR19R20XS(O)2NR16R17, -(CR19R20)nS(O)(OR16), -(CR19R20X S(O)2(OR16), -(CR19R20X SC(=Y')R16, -(CR19R20X SC(=Y')OR16, -(CR19R20X SC(=Y')NR16R17, and R21; and all other variables are as defined in Formula I or I-a to I-i, or as defined above. [00106] In another embodiment of the present invention, W is:
Figure imgf000033_0001
; and all other variables are as defined in Formula I or I-a to I-i, or as defined above.
[00107] In an embodiment of the present invention, X1 is R11; and all other variables are as defined in Formula I, I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, or I-i, or as defined above. [00108] In another embodiment of the present invention, X1 is R11 wherein R11 is H; and all other variables are as defined in Formula I, I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, or I-i, or as defined above.
[00109] In another embodiment of the present invention, X1 is R11 wherein R11 is Ci-
Ci2 alkyl (e.g., Ci-C6 alkyl) substituted with one or more groups independently selected from halo, CN, CF3, -OCF3, -NO2, oxo, -Si(Ci-C6 alkyl), -(CR19R20X C(=Y')R16, -(CR19R20)nC(=Y')OR16, -(CR19R20χC(=Y')NR16R17, -(CR19R20XNR16R17, -(CR19R20XOR16, -(CR19R20X-SR16, -(CR19R20)nNR16C(=Y')R17, -(CR19R20X NR16C(=Y')OR17, -(CR19R20X NR18C(=Y')NR16R17, -(CR19R20XNR17SO2R16, -(CR19R20)nOC(=Y')R16, -(CR19R20)nOC(=Y')OR16, -(CR19R20)nOC(=Y')NR16R17, -(CR19R20)nOS(O)2(OR16), -(CR19R20)nOP(=Y')(OR16)(OR17), -(CR19R20)nOP(OR16)(OR17), -(CR19R20)nS(O)R16, -(CR19R20)nS(O)2R16, -(CR19R20)n S(O)2NR16R17, -(CR19R20)nS(O)(OR16), -(CR19R20)nS(O)2(OR16), -(CR19R20)nSC(=Y')R16, -(CR19R20)nSC(=Y')OR16, -(CR19R20)nSC(=Y')NR16R17, and R21; and all other variables are as defined in Formula I, I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, or I-i, or as defined above. [00110] In another embodiment of the present invention, X1 is
Figure imgf000034_0001
Figure imgf000035_0001
other variables are as defined in Formula I, I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, or I-i, or as defined above.
[00111] In another embodiment of the present invention, X1 is
Figure imgf000035_0002
all other variables are as defined in Formula I, I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, or I-i, or as defined above.
[00112] In another embodiment of the present invention, X1 is -S(O)2R11, and all other variables are as defined in Formula I, I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, or I-i, or as defined above. [00113] In another embodiment of the present invention, X1 is -S(O)2R11 wherein R11 is H or methyl; and all other variables are as defined in Formula I, I-a, I-b, I-c, I-d, I-e, I-f, I- g, I-h, or I-i, or as defined above.
[00114] In an embodiment of the present invention, W is -OR11 (i.e., Formula III-a, III- b, III-c, Ill-d, Ill-e, III-f, III-g, III-h, or III-i) wherein R11 of W is H or Ci-Ci2 alkyl; and all other variables are as defined above.
[00115] In another embodiment of the present invention, W is -OR11 (i.e., Formula III- a, Ill-b, III-c, III-d, III-e, III-f, III-g, III-h, or III-i) wherein R11 of W is H; and all other variables are as defined above.
[00116] In another embodiment of the present invention, W is -OR11 (i.e., Formula III- a, Ill-b, III-c, III-d, III-e, III-f, III-g, III-h, or III-i) wherein R11 of W is Ci-C6 alkyl; and all other variables are as defined above.
[00117] In an embodiment of the present invention, X2 is aryl (e.g., phenyl), wherein said aryl is optionally substituted with one or more groups independently selected from halo, CN, CF3, -OCF3, -NO2, oxo, -Si(Ci-C6 alkyl), -(CR19R20)n C(=Y')R16, -(CR19R20)nC(=Y')OR16, -(CR19R20)nC(=Y')NR16R17, -(CR19R20)nNR16R17, -(CR19R20)nOR16, -(CR19R20)n-SR16, -(CR19R20)nNR16C(=Y')R17, -(CR19R20)n NR16C(=Y')OR17, -(CR19R20)nNR18C(=Y')NR16R17, -(CR19R20)nNR17SO2R16, -(CR19R20)nOC(=Y')R16, -(CR19R20)nOC(=Y')OR16, -(CR19R20)nOC(=Y')NR16R17, -(CR19R20)nOS(O)2(OR16), -(CR19R20)nOP(=Y')(OR16)(OR17), -(CR19R20)nOP(OR16)(OR17), -(CR19R20)nS(O)R16, -(CR19R20)nS(O)2R16, -(CR19R20)n S(O)2NR16R17, -(CR19R20)nS(O)(OR16), -(CR19R20)nS(O)2(OR16), -(CR19R20)n SC(=Y')R16, -(CR19R20)nSC(=Y')OR16, -(CR19R20)nSC(=Y')NR16R17, and R21; and all other variables are as defined in Formula I, I-a to I-i, II-a to II-i, or III-a to III-i, or as defined above. [00118] In another embodiment of the present invention, X2 is
Figure imgf000037_0001
all other variables are as defined in Formula I, I-a to I-i, II-a to II-i, or III-a to III-i, or as defined above.
[00119] In another embodiment of the present invention, X2 is
Figure imgf000038_0001
and all other variables are as defined in Formula I, I-a to I-i, II-a to II-i, or III-a to III-i, or as defined above.
[00120] In another embodiment of the present invention, X2 is
Figure imgf000038_0002
and all other variables are as defined in Formula I, I-a to I-i, II-a to II-i, or III-a to III-i, or as defined above.
[00121] In another embodiment of the present invention, X is
Figure imgf000039_0001
; and all other variables are as defined in Formula I, I-a to I-i, II-a to II-i, or III-a to III-i, or as defined above. [00122] In another embodiment of the present invention, X2 is
Figure imgf000039_0002
; and all other variables are as defined in Formula I, I-a to I-i, II-a to II-i, or III- a to III-i, or as defined above.
[00123] In another embodiment of the present invention, X2 is Cβ-Cio aryl substituted with Ci -C4 alkyl; and all other variables are as defined in Formula I, I-a to I-i, II-a to II-i, or III-a to III-i, or as defined above. [00124] In another embodiment of the present invention, X2 is
Figure imgf000039_0003
heterocyclyl . and all other variabies are as defined in Formula I, I-a to I-i, II-a to II-i, or III-a to III-i, or as defined above. [00125] In another embodiment of the present invention, X is
Figure imgf000040_0001
all other variables are as defined in Formula I, I-a to I-i, II-a to II-i, or III-a to III-i, or as defined above.
[00126] In another embodiment of the present invention, X2 is
Figure imgf000040_0002
and all other variables are as defined in Formula I, I-a to I-i, II-a to II-i, or III-a to III-i, or as defined above.
[00127] In another embodiment of the present invention, X is carbocyclyl (e.g., C4-C6 carbocyclyl) or heterocyclyl (e.g., 4- to 6-membered heterocyclyl), wherein said carbocyclyl or heterocyclyl is optionally substituted with one or more groups independently selected from halo, CN, CF3, -OCF3, -NO2, oxo, -Si(Cj-C6 alkyl), -(CR19R20)nC(=Y')R16, -(CR19R20)nC(=Y')OR16, -(CR19R20)nC(=Y')NR16R17, -(CR19R20)nNR16R17, -(CR19R20)nOR16, -(CR19R20)n-SR16, -(CR19R20)n NR16C(=Y')R17, -(CR19R20)n NR16C(=Y')OR17, -(CR19R20)n NR18C(=Y')NR16R17, -(CR19R20)nNR17SO2R16, -(CR19R20)nOC(=Y')R16, -(CR19R20)nOC(=Y')OR16, -(CR19R20)nOC(=Y')NR16R17, -(CR19R20)nOS(O)2(OR16), -(CR19R20)nOP(=Y')(OR16)(OR17), -(CR19R20)nOP(OR16)(OR17), -(CR19R20)nS(O)R16, -(CR19R20)nS(O)2R16, -(CR19R20)nS(O)2NR16R17, -(CR19R20)nS(O)(OR16), -(CR19R20)n S(O)2(OR16), -(CR19R20)nSC(=Y')R16, -(CR19R20)nSC(=Y')OR16, -(CR19R20X SC(=Y')NR16R17, and R21; and all other variables are as defined in Formula I, I-a to I-i, II-a to II-i, or III-a to III-i, or as defined above. [00128] In another embodiment of the present invention, X2 is C4-C6 carbocyclyl wherein said carbocyclyl is substituted with -C(=Y')R16; and all other variables are as defined in Formula I, I-a to I-i, II-a to II-i, or III-a to III-i, or as defined above. [00129] In another embodiment of the present invention, X is
Figure imgf000041_0001
; ancι an other variables are as defined in Formula I, I-a to I-i, II-a to II-i, or III-a to III-i, or as defined above.
[00130] Another embodiment of the present invention includes compounds described in EXAMPLES 5-159 and compounds below:
Figure imgf000042_0001
[00131] The present compounds are prepared according to the procedures described below in the schemes and examples or by methods known in the art. The starting materials and various intermediates may be obtained from commercial sources, prepared from commercially available compounds, or prepared using well known synthetic methods (for example, those described in WO02/06213, WO 03/077855 and WO03/077914). [00132] For example, 5-azabenzofurans of Formula (I-b), (II-b) or (III-b) may be prepared using the synthetic routes outlined in Schemes 1, 2 and 3. Scheme 1
heat
but is not limited to, and functionalised
Figure imgf000043_0001
[00133] Compounds of formula (IV) may be prepared using published methods described in the literature. They may be reacted with methylglycolate or ethylglycolate in the presence of a base, such as sodium hydride, in a suitable solvent, such as NN- dimethylformamide or 1 ,2-dimethoxyethane, at a temperature of from -5O0C to room temperature, to obtain compounds of formula (VI).
[00134] Compounds of formula (VI) may be converted to compounds of formula (VII) by reaction with a halogenating agent such as phosphorus oxybromide, neat or in a suitable solvent such as toluene, at a temperature of from room temperature to 14O0C. Alternatively, compounds of formula (VI) may be reacted with nonafluorobutane sulphonyl fluoride in the presence of a base such as diisopropylethylamine and a catalyst such as NN-dimethyl-4- aminopyridine, in a solvent such as dichloromethane at room temperature, with N- phenyltrifluoromethanesulfonimide in the presence of a base such as diisopropylethylamine, in a suitable solvent such as 1,2-dimethoxyethane at a temperature from room temperature to the reflux temperature of the solvent. In addition, compounds of formula (VI) may be treated with trifluoromethanesulphonic acid anhydride in the presence of a base such as pyridine in a solvent such as dichloromethane at a temperature of from -2O0C to ambient temperature. [00135] Compounds of formula (VIII) may be obtained from compounds of formula
(VII) by reaction with an aniline (incorporating appropriate substituents Rl), in the presence of a catalyst such as tris(dibenzylideneacetone)dipalladium (0) or palladium acetate, a base such as potassium phosphate, sodium tert-butoxide, l,8-diazabicyclo[5.4.1]undec-7-ene or cesium carbonate, a ligand such as 9,9'-dimethyl-4,5-bis(diphenylphosphino)xanthene, 2,2'- bis(diphenylphosphino)-l,l '-binaphthyl, 2-dicyclohexylphosphino-2'-(NN- dimethylamino)biphenyl, 2-dicyclohexylphosphino-2',6'-(dimethoxy)biphenyl or tri-butyl- phosphine in a suitable solvent such as toluene, 1,2-dimethoxyethane, tetrahydrofuran or dioxane, at a temperature of from room temperature to the reflux temperature of the solvent, or under microwave irradiation at a temperature of from 7O0C to 15O0C.
[00136] Alternatively compounds of formula (VIII) can be obtained from compounds of formula (VI) by reaction with compounds of formula (IX) (prepared using published methods described in the literature), in a suitable solvent such as toluene or 1,2- dimethoxyethane, at a temperature of from room temperature to the reflux temperature of the solvent, or under microwave irradiation at a temperature of from 1000C to 18O0C. [00137] Compounds of formula (X) can be obtained from compounds of formula
(VIII) by reaction with a base such as sodium hydroxide in a protic solvent such as ethanol or methanol, at a temperature of from room temperature up to reflux temperature. Compounds of formula (X) can be reacted with a functionalised hydroxylamine of formula (XII) (commercially available or prepared according to Scheme 8) or an amine, and a suitable coupling agent, such as O-(7-aza-benzo-triazol- l-yl)-N,N5N5 N5 -tetra-methyluronium hexafluoro-phosphate, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride or NN'-dicyclohexylcarbodiimide in the presence of N-hydroxy-l,2,3-benzotriazole, in the presence of a suitable base such as diisopropylethylamine or triethylamine in an inert solvent, such as tetrahydrofuran, NN-dimethylformamide, or dichloromethane at a temperature of about room temperature, to obtain the compounds of formula (XI). Alternatively, compounds of formula (XI) can be obtained directly from compounds of formula (VIII) by reaction with an amine or hydroxylamine DΝHR in the presence of a Lewis acid such as trimethyl aluminium in a solvent such as DCM, at a temperature of from room temperature up to reflux temperature.
Alternatively, compounds of formula (VIII) can be prepared from compounds of formula
(XIII), according to Scheme 2.
Scheme 2
X CN
(XIII)
R' is Me or Et Base, Solvent (XVI) or lower alkyl
X is halogen
(a) Y is H, (b) Y is halogen leaving group that is n= 0-5
Figure imgf000045_0001
[00138] Compounds of formula (XIII) may be prepared using published methods described in the literature. Compounds of general formula (XIV) can be prepared from compounds of formula (XIII) using methods described above for the preparation of compounds of formula (VI) from compounds of formula (IV).
[00139] Compounds of formula (VIII) may be obtained from compounds of formula
(XIV) by reaction with compounds of formula (XV) (incorporating appropriate substituents Rl), using methods described above for the preparation of compounds of formula (VIII) from compounds of formula (VI). Alternatively, compounds of formula (VIII) may be obtained from compounds of formula (XIV) by reaction with compounds of formula (XVI) (incorporating appropriate substituents Rl), in the presence of a base such as sodium hydride or lithium hexamethyldisilazane, in a suitable solvent such as tetrahydrofuran or N, N- dimethylformamide, at a temperature of from room temperature to 15O0C. [00140] Alternatively, compounds of formula (X) can also be prepared from compounds of formula (VII) according to Scheme 3_.
Scheme 3
Figure imgf000046_0001
[00141] Compounds of formula (VII) can be converted to compounds of formula
(XVII) using methods described above for the preparation of compounds of formula (X) from compounds of formula (VIII).
[00142] Compounds of formula (XVII) can be coupled to amines such as 2-amino-2- methyl-1-propanol using methods described above for the preparation of compounds of formula (XI) from compounds of formula (X), followed by reaction with an agent such as thionyl chloride or phosphorus oxychloride, neat or in a suitable solvent such as dichloromethane, chloroform or diethyl ether, at a temperature of from room temperature to reflux of the solvent, to afford compounds of formula (XVIII). [00143] Compounds of formula (XIX) may be obtained from compounds of formula
(XVIII) by reaction with an aniline (incorporating appropriate substituents Rl), in the presence of a catalyst such as tris(dibenzylideneacetone)dipalladium (0) or palladium acetate, a base such as potassium phosphate, sodium tert-butoxide, l,8-diazabicyclo[5.4.1]undec-7- ene or cesium carbonate, a ligand such as 9,9'-dimethyl-4,5-bis(diphenylphosphino)xanthene, 2,2'-bis(diphenylphosphino)-l,l '-binaphthyl, 2-dicyclohexylphosphino-2'-(N,N- dimethylamino)biphenyl, 2-dicyclohexylphosphino-2',6'-(dimethoxy)biphenyl or tri-butyl- phosphine in a suitable solvent such as toluene, 1, 2 -dimethoxy ethane, tetrahydrofuran or dioxane, at a temperature of from room temperature to the reflux temperature of the solvent, or under microwave irradiation at a temperature of from 7O0C to 15O0C.
[00144] Alternatively, compounds of formula (XIX) may be obtained from compounds of formula (XVIII) by reaction with anilines (incorporating appropriate substituents Rl), in the presence of a base such as sodium hydride or lithium hexamethyldisilazane, in a suitable solvent such as tetrahydrofuran or N,N-dimethylformamide, at a temperature of from room temperature to 15O0C.
[00145] Compounds of formula (X) may be obtained from compounds of formula
(XIX) by reaction with an acid such as hydrogen chloride, or acetic acid in a suitable solvent such as water, at a temperature of from room temperature to reflux of the solvent. [00146] 6-Azabenzofurans of Formula I-c, II-c or III-c may be prepared using the synthetic routes outlined in Scheme 4.
Scheme 3
1 ) POX3, solvent, heat solvent heat
optional
Base
not limited to, functionalised
Figure imgf000047_0001
heat
[00147] Compounds of formula (XX) may be prepared using published methods described in the literature. They may be reacted with methylglycolate or ethylglycolate in the presence of a phosphine such as triphenyl phosphine, an alkyl-azodicarboxylate such as diethyl azodicarboxylate or diisopropyl azodicarboxylate, in an aprotic solvent, such as tetrahydrofuran or diethyl ether, at a temperature of from room temperature to reflux of the solvent, to obtain compounds of formula (XXI).
[00148] Compounds of formula (XXI) may be reacted in the presence of a base, such as sodium hydride, in a suitable solvent, such as N,N-dimethylformamide or 1,2- dimethoxyethane, at a temperature of from -5O0C to room temperature, to obtain compounds of formula (XXII).
[00149] Compounds of formula (XXII) may be converted to compounds of formula
(XXIII) by reaction with a halogenating agent such as phosphorus oxybromide, neat or in a suitable solvent such as toluene, at a temperature of from room temperature to 14O0C. Alternatively, compounds of formula (XXII) may be reacted with nonafluorobutane sulphonyl fluoride in the presence of a base such as diisopropylethylamine and a catalyst such as N,N-dimethyl-4-aminopyridine, in a solvent such as dichloromethane at room temperature, with N-phenyltrifluoromethanesulfonimide in the presence of a base such as diisopropylethylamine, in a suitable solvent such as 1, 2 -dimethoxy ethane at a temperature from room temperature to the reflux temperature of the solvent. In addition compounds of formula (VI) may be treated with trifluoromethanesulphonic acid anhydride in the presence of a base such as pyridine in a solvent such as dichloromethane at a temperature of from - 2O0C to ambient temperature.
Compounds of formula (XXIV) may be obtained from compounds of formula (XXIII) by reaction with an aniline (incorporating appropriate substituents Rl), in the presence of a catalyst such as tris(dibenzylideneacetone)dipalladium (0) or palladium acetate, a base such as potassium phosphate, sodium tert-butoxide, l,8-diazabicyclo[5.4.1]undec-7-ene or cesium carbonate, a ligand such as 9,9'-dimethyl-4,5-bis(diphenylphosphino)xanthene, 2,2'- bis(diphenylphosphino)-l,l '-binaphthyl, 2-dicyclohexylphosphino-2'-(NN- dimethylamino)biphenyl, 2-dicyclohexylphosphino-2',6'-(dimethoxy)biphenyl or tri-butyl- phosphine in a suitable solvent such as toluene, 1,2-dimethoxyethane, tetrahydrofuran or dioxane, at a temperature from room temperature to the reflux temperature of the solvent, or under microwave irradiation at a temperature of from 7O0C to 15O0C. [00150] Alternatively compounds of formula (XXIV) can be obtained from compounds of formula (XXII) by reaction with compounds of formula (IX) (prepared using published methods described in the literature), in a suitable solvent such as toluene or 1,2- dimethoxyethane, at a temperature from room temperature to the reflux temperature of the solvent, or under microwave irradiation at a temperature of from 1000C to 18O0C. [00151] Compounds of formula (XXVI) can be obtained from compounds of formula
(XXIV) by reaction with a base such as sodium hydroxide in a protic solvent such as ethanol or methanol, at a temperature from room temperature up to reflux temperature. [00152] Compounds of formula (XXVI) can be reacted with a functionalised hydroxylamine of formula (XII) (commercially available or prepared according to Scheme 8) or an amine, and a suitable coupling agent, such as 0-(7-aza-benzo-triazol-l-yl)-N,N,N,N- tetra-methyluronium hexafluoro-phosphate, N-(3 -dimethylaminopropyl)-N'- ethylcarbodiimide hydrochloride or NN'-dicyclohexylcarbodiimide in the presence of N- hydroxy- 1, 2,3 -benzotriazole, in the presence of a suitable base such as diisopropylethylamine or triethylamine in an inert solvent, such as tetrahydrofuran, N,N-dimethylformamide, or dichloromethane at a temperature of about room temperature, to obtain the compounds of formula (XXVII). Alternatively, compounds of formula (XXVI) can be obtained directly from compounds of formula (XXIV) by reaction with an amine or hydroxylamine DΝHR in the presence of a Lewis acid such as trimethyl aluminium, in a solvent such as DCM, at a temperature of from room temperature up to reflux temperature.
[00153] Furo[2,3-d]pyrimidines of Formula I-f, II-f or III-f may be prepared using the synthetic routes outlined in Scheme 5. Scheme 4
HO CO2R'
^- Ν (XXVIII)
heat
but is not limited to, and functionalised
Figure imgf000049_0001
[00154] Compounds of formula (XXVIII) may be prepared according to methods described in the literature. They may reacted with a halogenating agent such as phosphorus oxychloride, neat or in a suitable solvent such as toluene, at a temperature from room temperature to reflux, to provide compounds of formula (XXIX).
Compounds of formula (XXXVI) may be obtained from compounds of formula (XXIX) using similar methods to the ones described for the preparation of compounds of formula (XI) from compounds of formula (IV), as shown in Scheme 1. [00155] Furo[2,3-d]pyridazines of formula I-h, II-h and II-h and furo[3,2-c]pyridazines of formula I-g, II-g, and III-g may be prepared using the synthetic routes outlined in Scheme 6. Scheme 6
heat
but is not limited to, and functionalised
Figure imgf000050_0001
[00156] Compounds of formula (L) may be prepared according to methods described in the literature. Compounds of formula (LVI) may be obtained from compounds of formula (L) using similar methods to the ones described for the preparation of compounds of formula (XI) from compounds of formula (IV), as shown in Scheme 1. Alternatively, compounds of formula (LIV) may be prepared according to Scheme 7.
Scheme 7
A1= N A2= CY or A1= CY A2= N
Figure imgf000051_0001
Compounds of formula (LVIII) may be prepared using published methods described in the literature. Compounds of general formula (LIV) can be prepared from compounds of formula
(LIX) using methods described above for the preparation of compounds of formula (VIII) from compounds of formula (XIII).
[00157] Hydroxylamines of formula (XII) may be prepared using methods described in the literature or the synthetic route outlined in Scheme 8.
Scheme 8
R.
(XXXVI Acid
Figure imgf000051_0002
R~o'NVR1 <X""b)
R11
[00158] Primary or secondary alcohols of general formula (XXXVII) may be prepared using methods described in the literature. They may be reacted with N-hydroxy phthalimide using a phosphine and coupling reagent such as diethyl azodicarboxylate to provide compounds of general formula (XXXVIII). Compounds of general formula (XXXVIII) may be deprotected using hydrazine or methyl hydrazine to provide hydroxylamines of general formula (XII-a). Compounds of formula (XII-a) may be further modified by reductive amination with aldehydes or ketones using a reducing agent such as sodium triacetoxy borohydride, sodium cyanoborohydride, or borane-pyridine in a solvent such as dichloroethane at a temperature of from ambient temperature to reflux. In addition, compounds of formula (XII-a) may be further modified by alkylation with an alkyl halide in the presence of a base such as triethylamine, in a solvent such as dichloromethane, to provide hydroxylamines of general formula (XII-b).
[00159] Anilines of general formula (XXXIX) used in cross-coupling reactions described above may be prepared by using methods described in the literature or according to Scheme 9.
Scheme 9
Figure imgf000052_0001
Where R1 is an optional substituent group, n= 0-4
[00160] Substituted 4-chloro-nitro benzene may be reacted with hexamethyldisilane in a solvent such as xylene using a catalyst such as tetrakis(triphenylphosphine)palladium at a temperature of from room temperature to reflux. The nitro group may be reduced using methods described in the literature such as reaction under an atmosphere of hydrogen at a pressure of from 1 to 5 atmospheres in the presence of a catalyst such as palladium on carbon and in a solvent such as ethanol or ethyl acetate at room temperature. [00161] Trifluoromethanesulfonyl esters of general formula (XL) used in cross- coupling reactions described above may be prepared by using methods described in the literature or according to Scheme 10.
Scheme 10 ONf
Figure imgf000052_0002
group, n= 0-Λ
[00162] Halo phenols of general structure (XLI) may be reacted with two equivalents of alkylithium reagents such as n-butyl lithium in a solvent such as THF, followed by quenching with trialkylsilyl halide such as trimethylsilyl chloride to give trialkylsilyl phenols (XLII). Trialkylsilyl phenols may be further reacted using literature procedures to give trifluoromethane sulfonates or nonaflates of general structure (XL). [00163] It will be appreciated that where appropriate functional groups exist, compounds of formula (I), (II), (III) or any intermediates used in their preparation may be further derivatised by one or more standard synthetic methods employing substitution, oxidation, reduction, or cleavage reactions. Particular substitution approaches include conventional alkylation, arylation, heteroarylation, acylation, sulfonylation, halogenation, nitration, formylation and coupling procedures.
[00164] For example, aryl bromide or chloride groups may be converted to aryl iodides using a Finkelstein reaction employing an iodide source such as sodium iodide, a catalyst such as copper iodide and a ligand such as trans-N,N'-dimethyl-l,2-cyclohexane diamine in a solvent such as 1,4-dioxane and heating the reaction mixture at reflux temperature. Aryl trialkylsilanes may be converted to aryl iodides by treating the silane with an iodide source such as iodine monochloride in a solvent such as dichloromethane with or without Lewis acid such as silver tetrafluoroborate at a temperature from -4O0C to reflux. [00165] In a further example primary amine (-NH2) groups may be alkylated using a reductive alkylation process employing an aldehyde or a ketone and a borohydride, for example sodium triacetoxyborohydride or sodium cyanoborohydride, in a solvent such as a halogenated hydrocarbon, for example 1,2-dichloroethane, or an alcohol such as ethanol, where necessary in the presence of an acid such as acetic acid at around ambient temperature. Secondary amine (-NH-) groups may be similarly alkylated employing an aldehyde. [00166] In a further example, primary amine or secondary amine groups may be converted into amide groups (-NHCOR' or -NRCOR') by acylation. Acylation may be achieved by reaction with an appropriate acid chloride in the presence of a base, such as triethylamine, in a suitable solvent, such as dichloromethane, or by reaction with an appropriate carboxylic acid in the presence of a suitable coupling agent such HATU (O-(7- azabenzotriazol-l-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate) in a suitable solvent such as dichloromethane. Similarly, amine groups may be converted into sulphonamide groups (-NHSO2R' or -NR"SO2R') groups by reaction with an appropriate sulphonyl chloride in the presence of a suitable base, such as triethylamine, in a suitable solvent such as dichloromethane. Primary or secondary amine groups can be converted into urea groups (-NHCONR'R" or -NRCONR'R") by reaction with an appropriate isocyanate in the presence of a suitable base such as triethylamine, in a suitable solvent, such as dichloromethane. [00167] An amine (-NH2) may be obtained by reduction of a nitro (-NO2) group, for example by catalytic hydrogenation, using for example hydrogen in the presence of a metal catalyst, for example palladium on a support such as carbon in a solvent such as ethyl acetate or an alcohol e.g. methanol. Alternatively, the transformation may be carried out by chemical reduction using for example a metal, e.g. tin or iron, in the presence of an acid such as hydrochloric acid.
[00168] In a further example, amine (-CH2NH2) groups may be obtained by reduction of nitriles (-CN), for example by catalytic hydrogenation using for example hydrogen in the presence of a metal catalyst, for example palladium on a support such as carbon, or Raney nickel, in a solvent such as an ether e.g. a cyclic ether such as tetrahydrofuran, at a temperature from -78°C to the reflux temperature of the solvent.
[00169] In a further example, amine (-NH2) groups may be obtained from carboxylic acid groups (-CO2H) by conversion to the corresponding acyl azide (-CON3), Curtius rearrangement and hydrolysis of the resultant isocyanate (-N=C=O).
[00170] Aldehyde groups (-CHO) may be converted to amine groups (-CH2NR5R")) by reductive amination employing an amine and a borohydride, for example sodium triacetoxyborohydride or sodium cyanoborohydride, in a solvent such as a halogenated hydrocarbon, for example dichloromethane, or an alcohol such as ethanol, where necessary in the presence of an acid such as acetic acid at around ambient temperature.
[00171] In a further example, aldehyde groups may be converted into alkenyl groups (-
CH=CHR') by the use of a Wittig or Wadsworth-Emmons reaction using an appropriate phosphorane or phosphonate under standard conditions known to those skilled in the art.
[00172] Aldehyde groups may be obtained by reduction of ester groups (such as -
CO2Et) or nitriles (-CN) using diisobutylaluminium hydride in a suitable solvent such as toluene. Alternatively, aldehyde groups may be obtained by the oxidation of alcohol groups using any suitable oxidising agent known to those skilled in the art.
[00173] Ester groups (-CO2R') may be converted into the corresponding acid group (-
CO2H) by acid- or base-catalused hydrolysis, depending on the nature of R. If R is t-butyl, acid-catalysed hydrolysis can be achieved for example by treatment with an organic acid such as trifluoroacetic acid in an aqueous solvent, or by treatment with an inorganic acid such as hydrochloric acid in an aqueous solvent.
[00174] Carboxylic acid groups (-CO2H) may be converted into amides (CONHR' or -
CONR'R") by reaction with an appropriate amine in the presence of a suitable coupling agent, such as HATU, in a suitable solvent such as dichloromethane. [00175] In a further example, carboxylic acids may be homologated by one carbon (i.e
-CO2H to -CH2CO2H) by conversion to the corresponding acid chloride (-COC1) followed by Arndt-Eistert synthesis.
[00176] In a further example, -OH groups may be generated from the corresponding ester (e.g. -CO2R'), or aldehyde (-CHO) by reduction, using for example a complex metal hydride such as lithium aluminium hydride in diethyl ether or tetrahydrofuran, or sodium borohydride in a solvent such as methanol. Alternatively, an alcohol may be prepared by reduction of the corresponding acid (-CO2H), using for example lithium aluminium hydride in a solvent such as tetrahydrofuran, or by using borane in a solvent such as tetrahydrofuran. [00177] Alcohol groups may be converted into leaving groups, such as halogen atoms or sulfonyloxy groups such as an alkylsulfonyloxy, e.g. trifluoromethylsulfonyloxy or arylsulfonyloxy, e.g. p-toluenesulfonyloxy group using conditions known to those skilled in the art. For example, an alcohol may be reacted with thioyl chloride in a halogenated hydrocarbon (e.g. dichloromethane) to yield the corresponding chloride. A base (e.g. triethylamine) may also be used in the reaction.
[00178] In another example, alcohol, phenol or amide groups may be alkylated by coupling a phenol or amide with an alcohol in a solvent such as tetrahydrofuran in the presence of a phosphine, e.g. triphenylphosphine and an activator such as diethyl-, diisopropyl, or dimethylazodicarboxylate. Alternatively alkylation may be achieved by deprotonation using a suitable base e.g. sodium hydride followed by subsequent addition of an alkylating agent, such as an alkyl halide.
[00179] Aromatic halogen substituents in the compounds may be subjected to halogen- metal exchange by treatment with a base, for example a lithium base such as n-butyl or t- butyl lithium, optionally at a low temperature, e.g. around -78°C, in a solvent such as tetrahydrofuran, and then quenched with an electrophile to introduce a desired substituent. Thus, for example, a formyl group may be introduced by using N,N-dimethylformamide as the electrophile. Aromatic halogen substituents may alternatively be subjected to metal (e.g. palladium or copper) catalysed reactions, to introduce, for example, acid, ester, cyano, amide, aryl, heteraryl, alkenyl, alkynyl, thio- or amino substituents. Suitable procedures which may be employed include those described by Heck, Suzuki, Stille, Buchwald or Hartwig. [00180] Aromatic halogen substituents may also undergo nucleophilic displacement following reaction with an appropriate nucleophile such as an amine or an alcohol. Advantageously, such a reaction may be carried out at elevated temperature in the presence of microwave irradiation.
[00181] The compounds of the present invention are tested for their capacity to inhibit
MEK activity and activation (primary assays) and for their biological effects on growing cells (secondary assays) as described below. The compounds having IC50 of less than 10 μM (more preferably less than 5 μM, even more preferably less than 1 μM, most preferably less than 0.5 μM) in the MEK activity assay of Example Ia or Ib, IC50 of less than 5 μM (more preferably less than 0.1 μM, most preferably less than 0.01 μM) in the MEK activation assay of Example 2, EC50 of less than 10 μM (more preferably less than 5 μM, most preferably less than 0.5 μM) in the cell proliferation assay of Example 3, and/or EC50 of less than 10 μM (more preferably less than 1 μM, most preferably less than 0.1 μM) in the ERK phosphorylation assay of Example 4, are useful as MEK inhibitors. [00182] The present invention includes a composition (e.g., a pharmaceutical composition) comprising a compound of Formula I (and/or solvates and salts thereof) and a carrier (a pharmaceutically acceptable carrier). The present invention also includes a composition (e.g., a pharmaceutical composition) comprising a compound of Formula I (and/or solvates and salts thereof) and a carrier (a pharmaceutically acceptable carrier), further comprising a second chemotherapeutic agent and/or a second anti-inflammatory agent such as those described herein. The present compositions are useful for inhibiting abnormal cell growth or treating a hyperproliferative disorder in a mammal (e.g., human). The present compositions are also useful for treating inflammatory diseases in a mammal (e.g., human). [00183] The present compounds and compositions are also useful for treating an autoimmune disease, destructive bone disorder, proliferative disorders, infectious disease, viral disease, fϊbrotic disease or neurodegenerative disease in a mammal (e.g., human). Examples of such diseases/disorders include, but are not limited to, diabetes and diabetic complications, diabetic retinopathy, retinopathy of prematurity, age-related macular degeneration, hemangioma, idiopathic pulmonary fibrosis, rhinitis and atopic dermatitis, renal disease and renal failure, polycystic kidney disease, congestive heart failure, neurofibromatosis, organ transplant rejection, cachexia, stroke, septic shock, heart failure, organ transplant rejection, Alzheimer's disease, chronic or neuropathic pain, and viral infections such as HIV, hepatitis (B) virus (HBV), human papilloma virus (HPV), cytomegalovirus (CMV), and Epstein-Barr virus (EBV). Chronic pain, for purposes of the present invention includes, but is not limited to, idiopathic pain, and pain associated with chronic alcoholism, vitamin deficiency, uremia, hypothyroidism, inflammation, arthritis, and post-operative pain. Neuropathic pain is associated with numerous conditions which include, but are not limited to, inflammation, postoperative pain, phantom limb pain, burn pain, gout, trigeminal neuralgia, acute herpetic and postherpetic pain, causalgia, diabetic neuropathy, plexus avulsion, neuroma, vasculitis, viral infection, crush injury, constriction injury, tissue injury, limb amputation, arthritis pain, and nerve injury between the peripheral nervous system and the central nervous system.
[00184] The present compounds and compositions are also useful for treating pancreatitis or kidney disease (including proliferative glomerulonephritis and diabetes- induced renal disease) in a mammal (e.g., human).
[00185] The present compounds and compositions are also useful for the prevention of blastocyte implantation in a mammal (e.g., human).
[00186] The present invention includes a method of inhibiting abnormal cell growth or treating a hyperproliferative disorder in a mammal (e.g., human) comprising administering to said mammal a therapeutically effective amount of a compound of Formula I (and/or solvates and salts thereof) or a composition thereof. Also included in the present invention is a method of treating an inflammatory disease in a mammal (e.g., human) comprising administering to said mammal a therapeutically effective amount of a compound of Formula I (and/or solvates and/or salts thereof) or a composition thereof.
[00187] The present invention includes a method of inhibiting abnormal cell growth or treating a hyperproliferative disorder in a mammal (e.g., human) comprising administering to said mammal a therapeutically effective amount of a compound of Formula I (and/or solvates and salts thereof) or a composition thereof, in combination with a second chemotherapeutic agent such as those described herein. The present invention also includes a method of treating an inflammatory disease in a mammal (e.g., human) comprising administering to said mammal a therapeutically effective amount of a compound of Formula I (and/or solvates and/or salts thereof) or a composition thereof, in combination with a second antiinflammatory agent such as those described herein.
[00188] The present invention includes a method of treating an autoimmune disease, destructive bone disorder, proliferative disorders, infectious disease, viral disease, fϊbrotic disease or neurodegenerative disease in a mammal (e.g., human) comprising administering to said mammal a therapeutically effective amount of a compound of Formula I (and/or solvates and salts thereof) or a composition thereof, and optionally further comprising a second therapeutic agent. Examples of such diseases/disorders include, but are not limited to, diabetes and diabetic complications, diabetic retinopathy, retinopathy of prematurity, age- related macular degeneration, hemangioma, idiopathic pulmonary fibrosis, rhinitis and atopic dermatitis, renal disease and renal failure, polycystic kidney disease, congestive heart failure, neurofibromatosis, organ transplant rejection, cachexia, stroke, septic shock, heart failure, organ transplant rejection, Alzheimer's disease, chronic or neuropathic pain, and viral infections such as HIV, hepatitis (B) virus (HBV), human papilloma virus (HPV), cytomegalovirus (CMV), and Epstein-Barr virus (EBV).
[00189] The present invention includes a method of treating pancreatitis or kidney disease (including proliferative glomerulonephritis and diabetes-induced renal disease) in a mammal (e.g., human) comprising administering to said mammal a therapeutically effective amount of a compound of Formula I (and/or solvates and salts thereof) or a composition thereof, and optionally further comprising a second therapeutic agent. [00190] The present invention includes a method for preventing of blastocyte implantation in a mammal (e.g., human) comprising administering to said mammal a therapeutically effective amount of a compound of Formula I (and/or solvates and salts thereof) or a composition thereof, and optionally further comprising a second therapeutic agent.
[00191] The present invention includes a method of using the present compounds for in vitro, in situ, and in vivo diagnosis or treatment of mammalian cells, organisms, or associated pathological conditions.
[00192] It is also believed that the compounds of the present invention can render abnormal cells more sensitive to treatment with radiation for purposes of killing and/or inhibiting the growth of such cells. Accordingly, this invention further relates to a method for sensitizing abnormal cells in a mammal (e.g., human) to treatment with radiation which comprises administering to said mammal an amount of a compound of Formula I (and/or solvates and salts thereof) or a composition thereof, which amount is effective is sensitizing abnormal cells to treatment with radiation.
[00193] Administration of the compounds of the present invention (hereinafter the
"active compound(s)") can be effected by any method that enables delivery of the compounds to the site of action. These methods include oral routes, intraduodenal routes, parenteral injection (including intravenous, subcutaneous, intramuscular, intravascular or infusion), topical, inhalation and rectal administration. [00194] The amount of the active compound administered will be dependent on the subject being treated, the severity of the disorder or condition, the rate of administration, the disposition of the compound and the discretion of the prescribing physician. However, an effective dosage is in the range of about 0.001 to about 100 mg per kg body weight per day, preferably about 1 to about 35 mg/kg/day, in single or divided doses. For a 70 kg human, this would amount to about 0.05 to 7 g/day, preferably about 0.05 to about 2.5 g/day. In some instances, dosage levels below the lower limit of the aforesaid range may be more than adequate, while in other cases still larger doses may be employed without causing any harmful side effect, provided that such larger doses are first divided into several small doses for administration throughout the day.
[00195] The active compound may be applied as a sole therapy or in combination with one or more chemotherapeutic agents, for example those described herein. Such conjoint treatment may be achieved by way of the simultaneous, sequential or separate dosing of the individual components of treatment.
[00196] The pharmaceutical composition may, for example, be in a form suitable for oral administration as a tablet, capsule, pill, powder, sustained release formulations, solution, suspension, for parenteral injection as a sterile solution, suspension or emulsion, for topical administration as an ointment or cream or for rectal administration as a suppository. The pharmaceutical composition may be in unit dosage forms suitable for single administration of precise dosages. The pharmaceutical composition will include a conventional pharmaceutical carrier or excipient and a compound according to the invention as an active ingredient. In addition, it may include other medicinal or pharmaceutical agents, carriers, adjuvants, etc. [00197] Exemplary parenteral administration forms include solutions or suspensions of active compounds in sterile aqueous solutions, for example, aqueous propylene glycol or dextrose solutions. Such dosage forms can be suitably buffered, if desired. [00198] Suitable pharmaceutical carriers include inert diluents or fillers, water and various organic solvents. The pharmaceutical compositions may, if desired, contain additional ingredients such as flavorings, binders, excipients and the like. Thus for oral administration, tablets containing various excipients, such as citric acid may be employed together with various disintegrants such as starch, alginic acid and certain complex silicates and with binding agents such as sucrose, gelatin and acacia. Additionally, lubricating agents such as magnesium stearate, sodium lauryl sulfate and talc are often useful for tableting purposes. Solid compositions of a similar type may also be employed in soft and hard filled gelatin capsules. Preferred materials, therefore, include lactose or milk sugar and high molecular weight polyethylene glycols. When aqueous suspensions or elixirs are desired for oral administration the active compound therein may be combined with various sweetening or flavoring agents, coloring matters or dyes and, if desired, emulsifying agents or suspending agents, together with diluents such as water, ethanol, propylene glycol, glycerin, or combinations thereof.
[00199] Methods of preparing various pharmaceutical compositions with a specific amount of active compound are known, or will be apparent, to those skilled in this art. For examples, see Remington's Pharmaceutical Sciences, Mack Publishing Company, Ester, Pa.,
15.sup.th Edition (1975).
[00200] EXAMPLES
Abbreviations
DBU l,8-Diazabicyclo[5.4.0]undec-7-ene
DCM Dichloromethane
DIAD diisopropyl azodicarboxylate
DIPEA Diisopropylethylamine
DMAP 4-Dimethylaminopyridine
DMF Dimethylformamide
EDCI 1 -Ethyl-3 -(3 ' -dimethylaminopropyl)carbodiimide
HATU O-(7-Azabenzotriazol- 1 -yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate HCl Hydrochloric acid
HM-Ν Isolute® HM-Ν is a modified form of diatomaceous earth that can efficiently absorb aqueous samples HOBt 1-Hydroxybenzotriazole
IMS industrial methylated spirits
ICl iodine monochloride
LDA Lithium diisopropylamide
MeOH Methanol
ΝaHCθ3 Sodium bicarbonate
NaOH Sodium hydroxide
Pd(PPli3)4 Tetrakis(triphenylphosphine)palladium(0)
Pd2dba3 Tris-(dibenzylideneacetone)dipalladium(0)
Si-SPE Pre-packed Isolute® silica flash chromatography cartridge Si-ISCO Pre-packed ISCO® silica flash chromatography cartridge
THF Tetrahydrofuran
Xantphos 9,9-Dimethyl-4,5-bis(diphenylphosphino)xanthene
[00201] General Experimental Conditions
[00202] 1H NMR spectra were recorded at ambient temperature using a Varian Unity
Inova (400MHz) spectrometer with a triple resonance 5mm probe. Chemical shifts are expressed in ppm relative to tetramethylsilane. The following abbreviations have been used: br = broad signal, s = singlet, d = doublet, dd = double doublet, t = triplet, q = quartet, m = multiple!
[00203] High Pressure Liquid Chromatography - Mass Spectrometry (LCMS) experiments to determine retention times (Rj) and associated mass ions were performed using one of the following methods.
[00204] Method A: Experiments performed on a Waters Micromass ZQ quadrupole mass spectrometer linked to a Hewlett Packard HPl 100 LC system with diode array detector. This system uses a Higgins Clipeus 5micron Cl 8 100 x 3.0mm column and a 1 ml / minute flow rate. The initial solvent system was 95% water containing 0.1% formic acid (solvent A) and 5% acetonitrile containing 0.1% formic acid (solvent B) for the first minute followed by a gradient up to 5% solvent A and 95% solvent B over the next 14 minutes. The final solvent system was held constant for a further 5 minutes.
[00205] Method B: Experiments performed on a Waters Platform LC quadrupole mass spectrometer linked to a Hewlett Packard HPl 100 LC system with diode array detector and 100 position autosampler using a Phenomenex Luna Cl 8(2) 30 x 4.6mm column and a 2 ml / minute flow rate. The solvent system was 95% water containing 0.1% formic acid (solvent A) and 5% acetonitrile containing 0.1% formic acid (solvent B) for the first 0.50 minutes followed by a gradient up to 5% solvent A and 95% solvent B over the next 4 minutes. The final solvent system was held constant for a further 0.50 minutes.
[00206] Method C: Experiments performed on a PE Sciex API 150 EX quadrupole mass spectrometer linked to a Shimadzu LC-IOAD LC system with diode array detector and 225 position autosampler using a Kromasil C18 50 x 4.6mm column and a 3 ml / minute flow rate. The solvent system was a gradient starting with 100% water with 0.05% TFA (solvent A) and 0% acetonitrile with 0.0375% TFA (solvent B), ramping up to 10% solvent A and 90% solvent B over 4 minutes. The final solvent system was held constant for a further 0.50 minutes. [00207] Method D: Experiments performed on a Shimadzu LCMS-2010A liquid chromatography mass spectrometer linked to a Shimadzu LC-IOAD VP LC system with diode array detector. Uses a Kromasil 100 5micron Cl 8 50 x 4.6 mm column and a 2.5 ml / minute flow rate. The initial solvent system was 100% water containing 0.05% trifluoroacetic acid (solvent A) and 0% acetonitrile containing 0.05% trifluoroacetic acid (solvent B), followed by a gradient up to 10% solvent A and 90% solvent B over 8 minutes. The final solvent system was held constant for a further 2 minutes.
[00208] Method E: Experiments performed on an Agilent Technologies liquid chromatography mass spectrometer linked to an Agilent Technologies Series 1100 LC system with diode array detector. Uses a Zorbax 3.5micron SB-C18 30 x 2.6 mm column and a 0.5 ml / minute flow rate. The initial solvent system was 95% water containing 0.05% trifluoroacetic acid (solvent A) and 5% acetonitrile containing 0.0375% trifluoroacetic acid (solvent B), followed by a gradient up to 5% solvent A and 95% solvent B over 9 minutes. The final solvent system was held constant for a further 1 minute.
[00209] Microwave experiments were carried out using a Personal Chemistry Emrys
Iniatiator™ or Optimizer™, which uses a single-mode resonator and dynamic field tuning, both of which give reproducibility and control. Temperature from 40-2500C can be achieved, and pressures of up to 20bar can be reached.
[00210] EXAMPLE Ia MEK Assay (MEK activity assay)
[00211] Constitutively activated human mutant MEKl expressed in insect cells is used as source of enzymatic activity at a final concentration in the kinase assay of 62.5nM. [00212] The assay is carried out for 30 minutes in the presence of 50μM ATP using recombinant GST-ERKl produced in E. CoIi as substrate. Phosphorylation of the substrate is detected and quantified using HTRF reagents supplied by Cisbio. These consist of an anti- GST antibody conjugated to allophycocyanin (XL665) and an anti-phospho (Thr202/Tyr204) ERK antibody conjugated to europium-cryptate. The anti-phospho antibody recognises ERKl dually phosphorylated on Thr202 and Tyr204. When both antibodies are bound to ERKl (i.e. when the substrate is phosphorylated), energy transfer from the cryptate to the allophycocyanin occurs following excitation at 340nm, resulting in fluorescence being emitted that is proportional to the amount of phosphorylated substrate produced. Fluorescence is detected using a multiwell fluorimeter.
[00213] Compounds are diluted in DMSO prior to addition to assay buffer and the final DMSO concentration in the assay is 1%. [00214] The IC50 is defined as the concentration at which a given compound achieves
50% inhibition of control. IC50 values are calculated using the XLfϊt software package (version 2.0.5).
[00215] EXAMPLE Ib MEK Assay (MEK activity assay)
[00216] Constitutively activated human mutant MEKl expressed in insect cells is used as source of enzymatic activity at a final concentration in the kinase assay of 15nM. [00217] The assay is carried out for 30 minutes in the presence of 50μM ATP using recombinant GST-ERKl produced in E. CoIi as substrate. Phosphorylation of the substrate is detected and quantified using HTRF reagents supplied by Cisbio. These consist of an anti- GST antibody conjugated to allophycocyanin (XL665) and an anti-phospho (Thr202/Tyr204) ERK antibody conjugated to europium-cryptate. These are used at a final concentration of 4μg/ml and 0.84μg/ml respectively. The anti-phospho antibody recognises ERKl dually phosphorylated on Thr202 and Tyr204. When both antibodies are bound to ERKl (i.e. when the substrate is phosphorylated), energy transfer from the cryptate to the allophycocyanin occurs following excitation at 340nm, resulting in fluorescence being emitted that is proportional to the amount of phosphorylated substrate produced. Fluorescence is detected using a multiwell fluorimeter.
[00218] Compounds are diluted in DMSO prior to addition to assay buffer and the final DMSO concentration in the assay is 1%.
[00219] The IC50 is defined as the concentration at which a given compound achieves
50% inhibition of control. IC50 values are calculated using the XLfit software package (version 2.0.5).
[00220] Compounds of Example 5-18, 20-102, 105-109, 111-118, 120-133, 136-149 and 151-160 exhibited an IC50 of less than 10 μM in the assay described either in Example Ia or Ib, most of these compounds exhibited an IC50 of less than 5 μM. [00221] EXAMPLE 2 bRaf Assay (MEK activation assay)
[00222] Constitutively activated bRaf mutant expressed in insect cells is used as source of enzymatic activity.
[00223] The assay is carried out for 30 minutes in the presence of 200μM ATP using recombinant GST-MEKl produced in E. CoIi as substrate. Phosphorylation of the substrate is detected and quantified using HTRF, and reagents are supplied by Cisbio. These consist of an anti-GST antibody conjugated to allophycocyanin (XL665) and an anti-phospho (Ser217/Ser221) MEK antibody conjugated to europium-cryptate. The anti-phospho antibody recognises MEK dually phosphorylated on Ser217 and Ser221 or singly phosphorylated on
Ser217. When both antibodies are bound to MEK (i.e. when the substrate is phosphorylated), energy transfer from the cryptate to the allophycocyanin occurs following excitation at
340nm, resulting in fluorescence being emitted that is proportional to the amount of phosphorylated substrate produced. Fluorescence is detected using a multiwell fluorimeter.
[00224] Compounds are diluted in DMSO prior to addition to assay buffer and the final DMSO concentration in the assay is 1%.
[00225] The IC50 is defined as the concentration at which a given compound achieves
50% inhibition of control. IC50 values are calculated using the XLfit software package
(version 2.0.5).
[00226] In this assay, compounds of Example 5-19 exhibited an IC50 of less than 5 μM.
[00227] EXAMPLE 3 Cell Proliferation Assay
[00228] Compounds are tested in a cell proliferation assay using the following cell lines:
[00229] HCT 116 human colorectal carcinoma (ATCC)
[00230] A375 human malignant melanoma (ATCC)
[00231] Both cell lines are maintained in DMEM/F12 (1: 1) media (Gibco) supplemented with 10% FCS at 370C in a 5% CO2 humidified incubator.
[00232] Cells are seeded in 96-well plates at 2,000 cells/well and after 24 hours they are exposed to different concentrations of compounds in 0.83% DMSO. Cells are grown for a further 72h, and an equal volume of CellTiter-Glo reagent (Promega) is added to each well.
This lyses the cells and generates a luminescent signal proportional to the amount of ATP released (and therefore proportional to the number of cells in the well) that can be detected using a multiwell luminometer.
[00233] The EC50 is defined as the concentration at which a given compound achieves
50% inhibition of control. EC50 values are calculated using the XLfit software package
(version 2.0.5).
[00234] In this assay, compounds of Example 5-13, 15-16, 18, 20-22, 24-25, 28, 31,
35, 38-39, 41, 109, 133-134, 138, 140-141 and 160 exhibited an EC50 of less than 10 μM in either one of the cell lines.
[00235] EXAMPLE 4 Phospho-ERK Cell-Based Assay
[00236] Compounds are tested in a cell-based phospho-ERK ELISA using the following cell lines:
[00237] HCT 116 human colorectal carcinoma (ATCC)
[00238] A375 human malignant melanoma (ATCC)
[00239] Both cell lines are maintained in DMEM/F12 (1: 1) media (Gibco) supplemented with 10% FCS at 370C in a 5% CO2 humidified incubator.
[00240] Cells are seeded in 96-well plates at 2,000 cells/well and after 24h they are exposed to different concentrations of compounds in 0.83% DMSO. Cells are grown for a further 2 h or 24h, fixed with formaldehyde (2% final) and permeabilised with methanol.
Following blocking with TBST-3% BSA, fixed cells are incubated with primary antibody
(anti-phospho ERK from rabbit) over-night at 40C. Cells are incubated with Propidium Iodide
(DNA fluorescent dye) and detection of cellular p-ERK is performed using an anti-rabbit secondary antibody conjugated to the fluorescent Alexa Fluor 488 dye (Molecular probes).
The fluorescence is analysed using the Acumen Explorer (TTP Labtech), a laser-scanning microplate cytometer, and the Alexa Fluor 488 signal is normalised to the PI signal
(proportional to cell number).
[00241] The EC50 is defined as the concentration at which a given compound achieves a signal half way between the baseline and the maximum response. EC50 values are calculated using the XLfit software package (version 2.0.5).
[00242] In this assay, compounds of Example 5-13, 15-16, 18, 20-26, 28-29, 31, 35,
38-39, 41-48, 50, 55, 59-61, 68, 70, 73-74, 76, 79, 81-84, 87, 91, 95, 99, 109, 111, 113, 117,
118, 120, 122-124, 126-127, 131, 133, 134, 138-141, 144, 147, 152, and 155-160 exhibited an EC50 of less than 10 μM in either one of the cell lines.
[00243] SYNTHESIS OF AZABENZOFURANYL CORES
[00244] Ethyl 3-(4-Bromo-2-fluoro-phenylamino)-furo[3,2-c1pyridine-2-carboxylate
Figure imgf000065_0001
[00245] Stepl: 4-Chloro-nicotinic acid
Figure imgf000065_0002
[00246] Following the procedures of Guillier et al (1995) J. Org. Chem. 60(2):292-6, to a cold (-780C) solution of LDA (21 ml, 1.6 M in hexanes, 33.3 mmol) in anhydrous THF (70 ml) was added 4-chloropyridine (5.0 g, 33.3 mmol) under an argon atmosphere. After 1 hour at -780C, the solution was rapidly poured onto a bed of solid CO2 contained within a 250 ml conical flask. After allowing to warm to ambient temperature the solution was quenched with water (30 ml). The volatile organic solvents were removed in vacuo and the remaining aqueous suspension was extracted with diethyl ether (3 x 100 ml). The aqueous phase was cooled to O0C and then adjusted to pH 4 by the addition of concentrated hydrochloric acid. The resultant precipitate was aged for 30 minutes then collected by filtration. The solid was washed with cold diethyl ether (10 ml) to afford the title compound as a white solid (3.2 g, 61%). [00247] Step 2: Ethyl 4-chloro-nicotinate
Figure imgf000066_0001
[00248] A suspension of 4-chloro-nicotinic acid (3.0 g, 19.0 mmol) in thionyl chloride
(50 ml) was heated under reflux for 90 minutes. After cooling to ambient temperature, the solution was concentrated to dryness and then azeotroped with toluene (2 x 50 ml) to afford a solid. The resultant solid was added in portions to a cooled (O0C) solution of ethanol (25 ml) and DIPEA (15 ml). The reaction was stirred at room temperature for 4 hours then concentrated in vacuo before water (75 ml) was added. The solution was extracted with ethyl acetate (2 x 75 ml) then the combined organic phases were dried over sodium sulfate then concentrated to give the title compound as a brown oil (3.3 g, 94%). IH NMR (CDCI3, 400MHz) 9.03 (s, IH), 7.58 (d, J = 5.4 Hz, IH), 7.41 (dd, J = 5.4 Hz, 0.5 Hz, IH), 4.45 (q, J = 7.3 Hz, 2H), 1.43 (t, J = 7.3 Hz, 3H). [00249] Step 3: Ethyl 3-hvdroxy-furor3,2-c"|pyridine-2-carboxylate
Figure imgf000066_0002
[00250] To a cooled (0 0C) solution of ethyl 4-chloro-nicotinate (910 mg, 4.9 mmol) and ethyl glycolate (0.48 ml, 5.1 mmol) in anhydrous DMF (17 ml), under a nitrogen atmosphere, was added sodium hydride (9.8 mmol, 60%, 392 mg). The reaction mixture was stirred for 16 hours (0 0C to room temperature), then acidified by the addition of acetic acid (1.2 ml), and subsequently concentrated to provide a residue. Water (23 ml) was added, and the mixture was stirred for 5 minutes, at which time the resulting brown precipitate was collected by filtration and washed with water 3 x 30 mL to give the title compound as a light brown solid. (875 mg, 86%). IH NMR (DMSO-D6, 400MHz) 9.18 (d, J = 1.2 Hz, IH), 8.60
(d, J = 6.0 Hz, IH), 7.66 (dd, J = 6.0 Hz, 0.8 Hz, IH), 4.32 (q, J = 7.2 Hz, 2H), 1.32 (t, J = 7.2
Hz, 3H). LCMS (method B): Rx = 1.42 min, M+H+ = 208.
[00251] Step 4: Ethyl 3-(trifluoromethanesulfonyloxy)-furor3,2-c1pyridine-2- carboxylate
Figure imgf000067_0001
[00252] A stirred solution of ethyl 3-hydroxy-furo[3,2-c]pyridine-2-carboxylate (3.15 g, 15.204 mmol), N,N-bis(trifluoromethylsulfonyl)aniline (10.08 g, 28.24 mmol) and NN- diisopropylethylamine (11.35 ml, 65.16 mmol) in dimethoxyethane (50 ml) was heated at 950C for 35 minutes. The reaction was then cooled to room temperature and concentrated under reduced pressure. The residue was then purified by flash chromatography (silica, 120 g column, ISCO, 45 mL/Min, 0-60% Ethyl acetate in hexane in 20 minutes) to afford the title compound as a pale yellow oil / white waxy solid (4. H g, 79.7%). IH ΝMR (CDCI3, 400MHz) 9.07 (s, IH), 8.75 (d, IH), 7.59 (d, IH), 4.54 (q, 2H), 1.47 (t, 3H). LCMS (5 min., method 2): Rx = 2.93 min, M+H+ = 339.6.
[00253] Step 5: Ethyl 3-(4-Bromo-2-fluoro-phenylamino)-furor3,2-clpyridine-2- carboxylate
[00254] A suspension of ethyl 3-(trifluoromethanesulfonyloxy)-furo[3,2-c]pyridine-2- carboxylate (4.11 g, 12.11 mmol), 4-bromo-2-fluoroaniline (3.76 g, 19.38 mmol), Pd2dba3 (925 mg, 1.01 mmol), Xantphos (591 mg, 1.02 mmol) and K3PO4 (4.95 g, 22.61 mmol) in toluene (60 ml) was degassed with bubbling nitrogen for 10 minutes and then heated to 1050C for 24 hours. The reaction mixture was then cooled to room temperature and diluted with ethyl acetate (100 ml). The resultant mixture was then filtered through celite 545 and the celite was washed with an additional 50 ml ethyl acetate. The filtrate was then concentrated and purified by flash chromatography (silica, 120 g column, ISCO, 45 mL/Min, 0-70% Ethyl acetate in hexane in 40 minutes) to afford the title compound as a white solid (2.96 g, 64.5%). IH ΝMR (CDCl3, 400MHz) 8.60 (m, 2H), 7.66 (s, IH), 7.50 (d,m IH), 7.39 (d,d, IH), 7.30 (d,m, IH), 7.16 (t, IH), 4.49 (q, 2H), 1.47 (t, 3H). LCMS (5 min., method 2): Rx = 2.47 min, M+H+ = 378.9. [00255] Ethyl 3-(2-fluoro-4-iodo-phenylamino)-furor2,3-c1pyridine-2-carboxylate
Figure imgf000068_0001
[00256] Step 1 : 3-Amino-isonicotinic acid
HON^O
(ym>
[00257] Following the procedures of Zhou et al (2001) Bioorg. Med. Chem. Lett.
9(8):2061-2071, bromine (1.22 ml, 23.9 mmol) was slowly added to a cooled (50C) 2.5N solution of NaOH (60 ml, 150 mmol) and after stirring for 5 minutes pyrrolo[3,4-c]pyridine- 1,3-dione (3.5 g, 23.6 mmol) was added. The temperature was raised to 8O0C and the mixture was stirred for 1 hour before being cooled to ambient temperature. Acetic acid (5.9 ml, 98.3 mmol) was cautiously added (N.B.: gas evolution) and the solution stirred for 10 minutes whereby a precipitate formed that was collected by filtration. The solid was washed with water (20 ml) and MeOH (20 ml) then dried to afford the title compound as a yellow solid (2.1 g, 64%).
[00258] Step 2: 3-Hydroxy-isonicotinic acid
HON^O
(V
[00259] To a suspension of 3-amino-isonicotinic acid (2.1 g, 15.2 mmol) in water (35 ml) was added concentrated sulphuric acid (1.5 ml). The solution was cooled to 50C and vigorously stirred before a solution of sodium nitrite (1.05 g, 15.2 mmol) in water (10 ml) was added. The suspension was slowly heated to 8O0C and held at this temperature for 15 minutes, then cooled to 650C and acetic acid (1.5 ml) was added. The pH of the solution was adjusted to pH 4.5 by the addition of concentrated ammonia solution (approximately 3.5 ml) then the mixture was placed in the refrigerator overnight. The resultant precipitate was collected by filtration, washed with water (20 ml) and dried under vacuum to afford the title compound as a yellow solid (1.85 g, 88%). IH NMR (d4-Me0H, 400 MHz) 8.37 (s, IH), 8.09 (d, J = 5.5 Hz, IH), 7.81 (d, J = 5.5 Hz, IH). [00260] Step 3: Ethyl 3-hydroxy-isonicotinate
Figure imgf000069_0001
[00261] 3-Hydroxy-isonicotinic acid (1.83 g, 13.2 mmol) was heated at reflux for 48 hours in a mixture of ethanol (40 ml) and concentrated sulphuric acid (1.0 ml). The mixture was cooled to room temperature and concentrated to give a residue. The residue was dissolved in water (10 ml) and neutralised by the addition of NaHCθ3 (approximately 2 g). The organic components were extracted with DCM (3 x 20 ml), and the combined organic extracts were dried over magnesium sulfate and concentrated to afford the title compound as a yellow oil that solidified on standing (1.87 g, 85%). IH NMR (d4-MeOH, 400 MHz) 10.40 (s, IH), 8.49 (s, IH), 8.21 (d, J = 5.20 Hz, IH), 7.62 (d, J = 5.20 Hz, IH), 4.47 (q, J = 6.44 Hz, 2H), 1.45 (t, J = 6.44 Hz, 3H). [00262] Step 4: Ethyl 3-ethoxycarbonylmethoxy-isonicotinate
Figure imgf000069_0002
[00263] To a cold (5 0C) solution of ethyl 3-hydroxy-isonicotinate (1.67 g, 1.0 mmol), ethyl glycolate (1.15 ml, 12.0 mmol) and triphenylphosphine (3.93 g, 15.0 mmol) in anhydrous THF (50 ml) was added diisopropylazodicarboxylate (2.94 ml, 15.0 mmol) dropwise. The reaction mixture was gradually warmed to ambient temperature then stirred for an additional 1 hour. The solution was concentrated and purified by flash chromatography (Si-SPE, pentane: diethyl ether, gradient 50:50 to 0: 100) to afford the title compound as a yellow oil (2.15 g, 85%). LCMS (method B): Rx = 2.69 min, M+H+ = 254. [00264] Step 5: Ethyl 3-hydroxy-furo|"2.3-c"lpyridine-2-carboxylate
Figure imgf000069_0003
[00265] A solution of ethyl S-ethoxycarbonylmethoxy-isonicotinate (2.1 g. 8.3 mmol) in THF (50 ml) was carefully added to a cold (O0C) solution of potassium tert-butoxide (966 mg, 8.6 mmol) in THF (20 ml). After 30 minutes the reaction mixture was quenched by the addition of acetic acid (10 ml). Evaporation of the solvents afforded a gum that was dissolved in ethyl acetate (50 ml) and washed with water (2 x 10 ml). The organic layer was isolated and dried over anhydrous sodium sulfate. The solution was concentrated to afford the title compound as a yellow solid (1.60 g, 94%). LCMS (method B): Rx = 1.89 min, M+H+ = 208. [00266] Step 6: Ethyl 3-(nonafluorobutane-l-sulfonyloxy)-furor2,3-c1pyridine-2- carboxylate
Figure imgf000070_0001
[00267] To a stirred suspension of ethyl 3-hydroxy-furo[2,3-c]pyridine-2-carboxylate
(1.16 g, 5.60 mmol) in DCM (15 ml) at O0C was added DIPEA (1.32 ml, 7.5 mmol) followed by nonafluorobutylsulfonyl fluoride (1.25 ml, 6.9 mmol). After 10 minutes the reaction was warmed to room temperature and stirred for an additional 20 hours. The reaction mixture was concentrated, the residue was dissolved in DCM (100 ml) and washed with water (50 ml) followed by IN NaOH solution (20 ml). The combined organic layer was isolated and dried over sodium sulfate and concentrated in vacuo. Purification by flash chromatography (Si- SPE, pentane: diethyl ether, gradient 80:20 to 50:50) afforded the title compound as a white solid (895 mg, 33%). LCMS (method B): Rx = 4.34 min, M+H+ = 490. [00268] Step 7: Ethyl 3-(4-bromo-2-fluoro-phenylamino)-furor2,3-c"|pyridine-2- carboxylate
Figure imgf000070_0002
[00269] A degassed solution of ethyl 3-(nonafluorobutane-l-sulfonyloxy)-furo[2,3- c]pyridine-2-carboxylate (838 mg, 1.71 mmol), 4-bromo-2-fluoroaniline (423 mg, 2.23 mmol), Pd2dba3 (78 mg, 0.09 mmol), Xantphos (99 mg, 0.17 mmol) and DBU (651 μl, 4.28 mmol) in toluene (3.3 ml) was subjected to microwave irradiation at 15O0C for 20 minutes.
The reaction mixture was concentrated and the resultant residue absorbed onto HM-N before being purified by flash chromatography (Si-SPE, pentane: diethyl ether, gradient 80:20 to
0: 100) to afford the title compound as a white solid (369 mg, 57%). LCMS (method B): Rx =
3.77 min, M+H+ = 380/382.
[00270] Step 8: Ethyl 3-(2-fluoro-4-iodo-phenylamino)-furor2.3-c1pyridine-2- carboxylate
[00271] A mixture of ethyl 3-(4-bromo-2-fluoro-phenylamino)-furo[2,3-c]pyridine-2- carboxylate (311 mg, 0.82 mmol), copper(I) iodide (8 mg, 0.04 mmol), sodium iodide (246 mg, 1.64 mmol) and trans-N,N'-dimethyl-l,2-cyclohexane diamine (13 μl, 0.08 mmol) in 1,4- dioxane (0.8 ml) was heated at 1150C for 26 hours under an argon atmosphere. Once the reaction mixture was cooled to room temperature, the mixture was concentrated then purified by flash chromatography (Si-SPE, EtOAc) to afford the title compound as a yellow oil (220 mg, 63%). LCMS (method B): Rx = 3.91 min, M+H+ = 427. [00272] Ethyl 3 -(2-fluoro-4-iodo-phenylamino)-furo[3.2-clpyridine-2 -carboxylate
Figure imgf000071_0001
[00273] Step 1: 2-Fluoro-4-Trimethylsilanyl-Phenylamine
Figure imgf000071_0002
[00274] 4-Chloro-2-fluoronitrobenzene (6.Og, 34.2 mmol) was added to a 100 mL round bottom flask, followed by hexamethyldisilane (18.9g, 129.0 mmol, 26.4 mL) and xylene (13 mL). The mixture was magnetically stirred while nitrogen was bubbled into the solution via glass pipette for 10 minutes or until the entire solid had dissolved. [00275] Tetrakis(triphenylphosphine)palladium(0) (1.0 g, 0.9 mmol) was added, the flask fitted with a reflux condenser, and the reaction was heated at reflux for 24-48 hours while a slow stream of nitrogen was passed through a rubber septum placed in the top of the condenser. After cooling to room temperature, the reaction mixture was diluted with ethyl ether (40 mL) and filtered through a plug of silica gel (30 mL of Siθ2/ethyl ether slurry packed into a 60 mL fritted glass funnel). The filter cake was washed with ethyl ether (60 mL) and the combined organics were concentrated in vacuo to an orange oil, which was purified by flash chromatography (250 mL silica gel, 98: 1: 1 hexane-CH2Cl2-ethyl ether), yielding the 2-fluoro-4-trimethylsilylnitrobenzene (5.45 g, 75%) as a yellow-orange oil. [00276] The 2-fluoro-4-trimethylsilylnitrobenzene (5.45g, 25.6 mmol) was then dissolved in ethanol (100 mL), transferred to a Parr shaker bottle, flushed with nitrogen, then charged with 10% Pd-C (0.4 g). The reaction mixture was hydrogenated for Ih on the Parr apparatus (45 psi H2), and then filtered through a plug of Celite. The filter cake was washed with ethanol, and the combined filtrates were concentrated. The resulting residue was purified by flash chromatography (250 mL silica gel, 95:5 hexane-ethyl ether), to afford the title compound as a tan oil (4.31 g, 92%).
[00277] Step 2: Ethyl 3-(4-Trimethylsilyl-2-fluoro-phenylamino)-furor3,2-clpyridine-
2-carboxylate
Figure imgf000072_0001
[00278] A suspension of ethyl 3-(trifluoromethanesulfonyloxy)-furo[3,2-c]pyridine-2- carboxylate (17.5 g, 51.58 mmol), 2-fluoro-4-trimethylsilanyl-phenylamine (10 g, 54.26 mmol), Pd2dba3 (2.98 g, 3.26 mmol), Xantphos (1.94 mg, 3.26 mmol) and K3PO4 (15.83 g, 72.34 mmol) in toluene (100 ml) was degassed with bubbling nitrogen for 10 minutes in a 300 mL pressure bottle and then heated to 1050C for 24 hours. The reaction mixture was then cooled to room temperature and diluted with ethyl acetate (200 ml). The resultant mixture was then filtered through celite 545 and the celite was washed with an additional 100 ml ethyl acetate. The filtrate was then concentrated and purified by flash column chromatography (silica, 0-55% Ethyl acetate in hexane) to afford the title compound as a yellow solid (17.9 g, 93.2 %). LCMS (method C): Rx = 2.47 min, M+H+ = 373. IH NMR (CDCl3, 400MHz) 8.66 (d, IH), 8.57 (d, IH), 7.52 (s, IH), 7.45 (d,d, IH), 7.30 (m, 2H), 4.50 (q, 2H), 1.49 (t, 3H). [00279] Step 3: Ethyl 3-(2-fluoro-4-iodo-phenylamino)-furor3,2-c1pyridine-2- carboxylate
[00280] 16.O g (72.49 mmol) of AgBF4 was quickly weighed out into a 100OmL round bottom flask and then capped with a rubber septum. The flask was then purged with dry N2 gas for 10 minutes, after which the flask was cooled to -500C, while maintaining an inert atmosphere. To this was added 300 ml dry dichloromethane and then the resultant mixture was stirred for 15 minutes at -500C under nitrogen. To the reaction mixture was then added 9.0 g (24.16 mmol) of ethyl 3-(4-Trimethylsilyl-2-fluoro-phenylamino)-furo[3,2-c]pyridine- 2-carboxylate in 75 ml dry dichloromethane and the mixture stirred for 30 mins at -500C under nitrogen. The color of the reaction was clear yellow. The reaction was then treated with 25 ml ICl (1.0 M in CH2Cl2, 25 mmol) dropwise with stirring over 30 minutes. Addition of ICl resulted in a precipitate (white/brown, the color of the reaction was yellow - localized red color on contact of ICl with the reaction, which turned yellow with white ppt). The reaction was stirred at -500C under nitrogen for 30 minutes. LC/MS showed that the reaction was complete. The reaction was then quenched at -500C by addition of 200 ml sat. Na2S2O3 solution, followed by 100 ml of water. The mixture was then transferred to a sep. funnel and shaken. The mixture was then filtered through filter paper. The black solid on the filter paper was further rinsed with dichloromethane and then discarded. The filtrate was then transferred to a separatory funnel. This was then quickly extracted with dichloromethane (3 X 100 ml). The combined dichloromethane layers were then washed with 170 mL 4M NH4OH solution in a sep. funnel. The dichloromethane layer was then separated and bubbled with Nitrogen to remove the ammonia. This was then dried with Magnesium sulfate, filtered and concentrated under reduced pressure to give a yellow solid. The solid was then powdered and triturated with ether (2 X 30 ml) and then dried under vacuum to give 8.90 g of the title product (yellow solid, 86.4%)). LCMS (method C): Rx = 2.47 min, M+H+ = 427. IH NMR (CDCl3, 400MHz) 8.64 (d, IH), 8.9 (d, IH), 7.66 (s, IH), 7.54 (d,d, IH), 7.46 (d,d,m, 2H), 7.13 (t, IH), 4.49 (q, 2H), 1.49 (t, 3H). [00281] Ethyl 3-(4-iodo-phenylamino)-furor3,2-clpyridine-2-carboxylate
Figure imgf000073_0001
[00282] Step 1: Ethyl 3-amino-furor3,2-clpyridine-2-carboxylate
Figure imgf000073_0002
[00283] To a stirred mixture of sodium hydride (60% suspension in mineral oil, 6.0 g,
150 mmol) in DMF (160 ml) at -1O0C under a nitrogen atmosphere, was added ethyl glycolate (14.5 ml, 150 mmol) over 5 minutes. After 35 minutes the reaction mixture was further cooled to -35°C and a solution of 4-chloronicotinonitrile (6.9 g, 50 mmol) in DMF (40 ml) added over 5 minutes. The reaction mixture was then allowed to warm gradually over 1.5 hours to -5°C, before being quenched with a solution of acetic acid: water (45 ml: 400 ml), and then extracted with ethyl acetate (2 x 200 ml). The separated aqueous phase was basified by addition of solid sodium bicarbonate, and extracted with ethyl acetate (3 x 200 ml). The combined organic extracts were washed with sodium bicarbonate solution (100 ml) and water (2 xlOO ml), then the organic phase was isolated, dried (MgSO4), filtered and evaporated in vacuo. Purification of the resultant residue by flash chromatography (Si-SPE, cyclohexane: ethyl acetate, gradient 60:40 to 0: 100 then ethyl acetate: methanol, 90: 10) afforded the title compound as a pale yellow solid (6.25g, 61%). LCMS (method B): Rx = 1.45 min, M+H+ = 207.
[00284] Step 2: Ethyl 3-(4-iodo-phenylamino)-furor3,2-c"|pyridine-2-carboxylate [00285] A degassed solution of ethyl 3-amino-furo[3,2-c]pyridine-2-carboxylate (206 mg, 1.0 mmol), 1,4-diiodobenzene (3.3 g, 10.0 mmol), Pd2dba3 (24 mg, 26 μmol), Xantphos (30 mg, 52 μmol) and potassium phosphate (424 mg, 2.0 mmol) in toluene (10 ml) was stirred and heated to 1050C under an argon atmosphere for 42 hours. The cooled reaction mixture was poured into aqueous ammonium chloride solution and extracted with ethyl acetate (3 x 70 ml). The combined extracts were washed with water (2 x 100 ml) followed by brine (50 ml) before the organic phase was isolated, dried (MgSO4), filtered and evaporated in vacuo. Purification of the resultant residue by flash chromatography (Si-SPE, cyclohexane: ethyl acetate, gradient 100:0 to 60:40) afforded the title compound as an off white solid (100 mg, 24%). LCMS (method B): Rx = 3.16 min, M+H+ = 409. [00286] Ethyl 3-(2-chloro-4-iodo-phenylamino)-furo[3.2-clpyridine-2-carboxylate
Figure imgf000074_0001
[00287] Step 1 : Ethyl 3-(4-Bromo-2-chloro-phenylamino)-furor3,2-clpyridine-2- carboxylate
Figure imgf000074_0002
[00288] A degassed solution of ethyl 3-(nonafluorobutane-l-sulfonyloxy)-furo[3,2- c]pyridine-2-carboxylate (500 mg, 1.02 mmol), 4-bromo-2-chloroaniline (275 mg, 1.33 mmol), Pd2dba3 (47 mg, 0.05 mmol), Xantphos (59 mg, 0.10 mmol) and DBU (388 μl, 2.56 mmol) in toluene (2.0 ml) was subjected to microwave irradiation at 15O0C for 10 minutes. The reaction mixture was concentrated and the resultant residue absorbed onto HM-N before being purified by flash chromatography (Si-SPE, cyclohexane: ethyl acetate, gradient 100:0 to 0: 100) to afford the title compound as a white solid (183 mg, 47%). LCMS (method B): Rx = 3.54 min, M+H+ = 395/397.
[00289] Step 2: Ethyl 3-(2-chloro-4-iodo-phenylamino)-furor3,2-c1pyridine-2- carboxylate
[00290] A mixture of ethyl 3-(4-bromo-2-chloro-phenylamino)-furo[3,2-c]pyridine-2- carboxylate (183 mg, 0.46 mmol), copper(I) iodide (4 mg, 0.02 mmol), sodium iodide (139 mg, 0.93 mmol) and trans-N,N'-dimethyl-l,2-cyclohexane diamine (7 μl, 0.04 mmol) in 1,4- dioxane (0.5 ml) was heated at 1150C for 44 hours under an argon atmosphere. The reaction mixture was cooled to room temperature, then additional copper(I) iodide (4 mg, 0.02 mmol) and trans-N,N'-dimethyl-l,2-cyclohexane diamine (7 μl, 0.04 mmol) were added and heating resumed at 1150C for 18 hours under an argon atmosphere. The reaction mixture was then cooled to room temperature, diluted with dichloromethane, and washed with a 10% solution of ammonia in water, water then brine. The organic extract was dried over sodium sulfate, filtered and concentrated in vacuo to give a residue that was purified by flash chromatography (Si-SPE, cyclohexane: ethyl acetate, gradient 100:0 to 0: 100) to afford the title compound as an off-white solid (115 mg, 57%). LCMS (method B): Rx = 3.97 min, M+H+ = 443. [00291] Ethyl 3-(2.6-difluoro-4-iodo-phenylamino)-furor3.2-clpyridine-2-carboxylate
Figure imgf000075_0001
[00292] Step 1 : Ethyl 3-(4-bromo-2,6-difluoro-phenylamino)-furor3,2-clpyridine-2- carboxylate
Figure imgf000075_0002
[00293] A degassed solution of ethyl 3-(nonafluorobutane-l-sulfonyloxy)-furo[3,2- c]pyridine-2-carboxylate (500 mg, 1.02 mmol), 4-bromo-2,6-difluoroaniline (277 mg, 1.33 mmol), Pd2dba3 (47 mg, 0.05 mmol), Xantphos (59 mg, 0.10 mmol) and DBU (388 μl, 2.56 mmol) in toluene (2.0 ml) was subjected to microwave irradiation at 15O0C for 10 minutes. The reaction mixture was concentrated and the resultant residue absorbed onto HM-N before being purified by flash chromatography (Si-SPE, cyclohexane: ethyl acetate, gradient 100:0 to 0: 100) to afford the title compound as a white solid (89 mg, 22%). LCMS (method B): Rx = 3.38 min, M+H+ = 397/399.
[00294] Step 2: Ethyl 3-(2.6-difluoro-4-iodo-phenylamino)-furo|"3.2-c]pyridine-2- carboxylate
[00295] A mixture of ethyl 3-(4-bromo-2,6-difluoro-phenylamino)-furo[3,2- c]pyridine-2-carboxylate (165 mg, 0.42 mmol), copper(I) iodide (4 mg, 0.02 mmol), sodium iodide (125 mg, 0.83 mmol) and trans-N,N'-dimethyl-l,2-cyclohexane diamine (7 μl, 0.04 mmol) in 1,4-dioxane (0.5 ml) was subjected to microwave irradiation at 18O0C for 15 minutes. Additional copper(I) iodide (4 mg, 0.02 mmol), sodium iodide (60 mg, 0.40 mmol) and trans-N,N'-dimetriyl-l,2-cyclohexane diamine (7 μl, 0.04 mmol) were added to the reaction mixture that was re-subjected to microwave irradiation at 18O0C for 15 minutes. The reaction mixture was diluted with dichloromethane and washed with a 10% solution of ammonia in water, water then brine. The organic extract was dried over sodium sulfate, filtered and concentrated to give a residue that was purified by flash chromatography (Si- SPE, cyclohexane: ethyl acetate, gradient 100:0 to 0: 100) to afford the title compound as an off-white solid (137 mg, 74%). LCMS (method B): Rx = 3.48 min, M+H+ = 445. [00296] Ethyl 3-(2.5-difluoro-4-iodo-phenylamino)-furo[3.2-clpyridine-2-carboxylate
Figure imgf000076_0001
[00297] Step 1 : Ethyl 3-(4-bromo-2,5-difluoro-phenylamino)-furor3,2-clpyridine-2- carboxylate
Figure imgf000076_0002
[00298] A degassed solution of ethyl 3-(nonafluorobutane-l-sulfonyloxy)-furo[3,2- c]pyridine-2-carboxylate (500 mg, 1.02 mmol), 4-bromo-2,5-difluoroaniline (277 mg, 1.33 mmol), Pd2dba3 (47 mg, 0.05 mmol), Xantphos (59 mg, 0.10 mmol) and DBU (388 μl, 2.56 mmol) in toluene (2.0 ml) was subjected to microwave irradiation at 15O0C for 10 minutes. The reaction mixture was concentrated and the resultant residue absorbed onto HM-N before being purified by flash chromatography (Si-SPE, cyclohexane: ethyl acetate, gradient 100:0 to 0: 100) to afford the title compound as a white solid (231 mg, 57%). LCMS (method B): Rx = 3.22 min, M+H+ = 397/399.
[00299] Step 2: Ethyl 3-(2,5-difluoro-4-iodo-phenylamino)-furor3,2-c"|pyridine-2- carboxylate
[00300] A mixture of ethyl 3-(4-bromo-2,5-difluoro-phenylamino)-furo[3,2- c]pyridine-2-carboxylate (222 mg, 0.56 mmol), copper(I) iodide (5 mg, 0.03 mmol), sodium iodide (168 mg, 1.12 mmol) and trans-N,N'-dimethyl-l,2-cyclohexane diamine (10 μl, 0.06 mmol) in 1,4-dioxane (0.5 ml) was heated at 11O0C for 18 hours. Additional copper(I) iodide (5 mg, 0.03 mmol), and trans-N,N'-dimethyl-l,2-cyclohexane diamine (10 μl, 0.06 mmol) were added to the reaction mixture that was re -heated at HO0C for 6 hours. The reaction mixture was cooled, diluted with dichloromethane, and washed with a 10% solution of ammonia in water, water then brine. The organic extract was dried over sodium sulfate, filtered and concentrated to give a residue that was purified by flash chromatography (Si- SPE, cyclohexane: ethyl acetate, gradient 100:0 to 0: 100) to afford the title compound as a white solid (170 mg, 68%). LCMS (method B): Rx = 3.30 min, M+H+ = 445. [00301] 7-Bromo-3-(2-fluoro-4-iodo-phenylamino)-furor3,2-clpyridine-2-carboxyric acid
Figure imgf000077_0001
[00302] Step 1: Ethyl 7-bromo-3-hvdroxy-furor3,2-c"|pyridine-2-carboxylate
Figure imgf000077_0002
[00303] To a solution of ethyl 4,5-dibromonicotinate (2.68 g, 8.67 mmol) and ethyl glycolate (0.90 g, 8.67 mmol) in DMF (25 ml) at O0C (ice/water), was added sodium hydride (1.04 g, 26 mmol, 60% oil dispersion). The reaction mixture was stirred at O0C for 15 min before being allowed to warm to room temperature for 2h. The reaction mixture was cooled to O0C before the addition of IM HCl (18 ml, 18 mmol). The precipitate was filtered and washed with water to afford the title compound as an off-white solid (2.35 g, 95%). LCMS (method B): Rx = 2.96 min, M+H+ = 285/287.
[00304] Step 2: 3.7-Dibromo-furo|"3.2-c1pyridine-2-carboxyric acid
Figure imgf000077_0003
[00305] A mixture of ethyl 7-bromo-3-hydroxy-furo[3,2-c]pyridine-2-carboxylate
(1.14 g, 4.0 mmol) and phosphorus oxybromide (5.6 g, 19.5 mmol) were heated at 14O0C for 2 hours. The reaction mixture was cooled to room temperature before the addition of crushed ice (ca 30 ml). The mixture was neutralised by the addition of solid NaOH before being adjusted to pH 3.0 by the careful addition of IM HCl. The resultant precipitate was filtered, and then washed with water followed by dichloromethane to afford the title compound as a white solid (1.2 g, 90%). LCMS (method B): Rx = 2.62 min, M+H+ = 320/322/324. [00306] Step 3: 3 J-Dibromo-furorS^-cipyridine^-carboxyric acid (2 -hydroxy- 1.1- dimethyl-ethyD-amide
Figure imgf000078_0001
[00307] A mixture of 3,7-dibromo-furo[3,2-c]pyridine-2-carboxylic acid (1.2 g, 3.74 mmol) and carbonyl diimidazole (0.85 g, 5.24 mmol) in acetonitrile (18 ml) were heated at 5O0C for 2 hours. A further portion of carbonyl diimidazole (0.035 g, 0.5 mmol) was added to the reaction mixture and heating at 5O0C continued for 1 hour. After cooling to ambient temperature 2-amino-2-methyl-propan-l-ol (0.30 ml, 3.13 mmol) was added to the solution. The reaction mixture was left to stand at room temperature for 19 hours then heated to 5O0C for 1 hour before being concentrated in vacuo. Purification of the resultant residue by flash chromatography (Si-SPE, cyclohexane: ethyl acetate, gradient 80:20 to 0: 100) afforded the title compound as a pale yellow solid (0.53 g, 72%). LCMS (method B): Rx = 2.57 min, M+H+ = 391/393/395.
[00308] Step 4: 3 J-Dibromo-2-(4.4-dimethyl-4.5-dihvdro-oxazol-2-yl)-furor3.2- cipyridine
Figure imgf000078_0002
[00309] To a solution of 3,7-dibromo-furo[3,2-c]pyridine-2-carboxylic acid (2- hydroxy-l,l-dimethyl-ethyl)-amide (0.53 g, 1.35 mmol) in dichloromethane (10 ml) was added thionyl chloride (0.25 ml, 3.43 mmol). The mixture was stirred at room temperature for Ih, then heated to reflux for 2h, before cooling to O0C. The mixture was neutralised with IM NaOH (15ml) and the aqueous layer was extracted with dichloromethane (2 x 15 ml). The organic layer was collected then dried over magnesium sulfate and concentrated in vacuo to afford a residue. Purification of the resultant residue by flash chromatography (Si-SPE, dichloromethane then ethyl acetate) afforded the title compound as a pale yellow gum (250 mg, 50%). LCMS (method B): Rx = 3.11 min, M+H+ = 373/375/377.
[00310] Step 5: r7-Bromo-2-(4.4-dimethyl-4.5-dihvdro-oxazol-2-vn-furor3.2- c1pyridin-3-yl1-(2-fmoro-4-iodo-phenvi)-amine
Figure imgf000079_0001
[00311] To a solution of 3,7-dibromo-2-(4,4-dimethyl-4,5-dihydro-oxazol-2-yl)- furo[3,2-c]pyridine (250 mg, 0.67 mmol) and 4-iodo-2-fluoroaniline (474 mg, 2 mmol) in THF (2ml) was added lithium hexamethyldisilazide solution in THF (2ml, IM solution). The reaction mixture was heated at 5O0C for 4 hours then cooled to room temperature and diluted with water (15 ml). The aqueous layer was extracted with dichloromethane (2 x 10 ml), and the combined organic extracts dried over MgSO4 and concentrated in vacuo to afford a residue. Purification of the resultant residue by flash chromatography (Si-SPE, cyclohexane: tert-butyl-methylether, gradient 1 : 1 to 1 :2) afforded the title compound as a light brown solid (150 mg, 42%). LCMS (method A): Rx = 13.97 min, M+H+ = 530/532. [00312] Step 6: 7-Bromo-3-(2-fluoro-4-iodo-phenylamino)-furo|"3.2-c]pyridine-2- carboxylic acid
Figure imgf000079_0002
[00313] A mixture of [7-bromo-2-(4,4-dimethyl-4,5-dihydro-oxazol-2-yl)-furo[3,2- c]pyridin-3-yl]-(2-fluoro-4-iodo-phenyl)-amine (110 mg, 0.2 mmol) and IM HCl (2 ml, 2 mmol) was heated at 1000C for 4 hours then cooled and concentrated in vacuo. The resultant residue was dissolved in methanol (3 ml) and 2.5M NaOH in methanol was added (0.4 ml, 1 mmol) followed by water (1 ml). The mixture was then heated at 750C for 1 hour before IM aqueous NaOH (1 ml, 1 mmol) was added and heating continued for 2 hours. The reaction mixture was concentrated in vacuo and the remaining aqueous layer was washed with ethyl acetate (2 x 2 ml). The aqueous layer was then acidified to pH 4 with IM HCl (~1.5 ml) and concentrated in vacuo to approximately half volume and allowed to stand at room temperature. The resultant precipitate was collected by filtration and washed with water (1 ml) followed by ethyl acetate (1 ml) to afford the title compound as a yellow / brown solid (66 mg, 69%). LCMS (method B): Rx = 3.34 min, M+H+ = 477/479.
[00314] Ethyl 5-(2-fluoro-4-iodo-phenylamino)-furor2,3-d1pyrimidine-6-carboxylate
Figure imgf000080_0001
[00315] Step 1 : Ethyl 4-hydroxy-pyrimidine-5-carboxylate
Figure imgf000080_0002
[00316] To a pre-formed solution of sodium (1.70 g, 73.9 mmol) in absolute ethanol
(300ml) was added 1,3,5-triazine (6.0 g, 74.1 mmol) and diethylmalonate (11.3 ml, 74.1 mmol). The reaction mixture was heated to reflux. After heating for 3h, the reaction mixture was cooled to room temperature and concentrated under reduced pressure to afford a residue. The residue was dissolved in water (300 ml), then cooled to 5°C and acidified by addition of hydrochloric acid (6 ml). The mixture was aged for 48 hours at 5°C and filtered. The resultant solid was washed with water, before being dried under reduced pressure to give the title compound as a beige solid (3.0g, 24%). 1H NMR (d6-DMSO, 400 MHz) 8.47 (s, IH), 8.37 (d, s, IH), 4.22 (q, J = 7.2 Hz, 2H), 1.26 (t, J = 7.2 Hz, 3H). [00317] Step 2: Ethyl 4-chloro-pyrimidine-5-carboxylate
Figure imgf000080_0003
[00318] To a suspension of ethyl 4-hydroxy-pyrimidine-5-carboxylate (3.0g, 17.6 mmol) in toluene (35ml) was added diisopropylethylamine (3.4 ml, 19.6mmol) and phosphorous oxychloride (1.8 ml, 19.6 mmol) dropwise under a nitrogen atmosphere. The reaction mixture was heated to 700C, and stirred for two hours then cooled to 5°C. A IM aqueous solution of sodium hydroxide (26 ml) was added and the mixture was diluted with water and extracted into ethyl acetate. The organic layer was washed with water, saturated sodium hydrogencarbonate, then dried over sodium sulfate, filtered and concentrated to give the title compound as a brown oil (2.56g, 77%). 1H NMR (CDCl3, 400 MHz) 9.13 (s, IH), 9.08 (s, IH), 4.47 (q, J = 6.9 Hz, 2H), 1.44 (t, J = 6.9 Hz, 3H). [00319] Step 3: Ethyl 4-ethoxycarbonylmethoxy-pyrimidine-5-carboxylate
Figure imgf000081_0001
[00320] To a suspension of sodium hydride (60% in mineral oil, 602 mg, 15.1 mmol) in anhydrous THF (55 mL) at 50C under a nitrogen atmosphere was added ethyl glycolate (1.6 ml, 16.5 mmol). The reaction mixture was stirred at 5°C for 30 minutes before a solution of ethyl 4-chloro-pyrimidine-5-carboxylate (2.56 g, 13.8 mmol) in anhydrous THF (20 ml) was added dropwise. The reaction mixture was stirred at 5°C for 30 minutes. Acetic acid (3 ml) was added to the reaction mixture that was then concentrated in vacuo. The resultant residue was dissolved in ethyl acetate and washed with water, then brine, before being dried over sodium sulfate and concentrated under reduced pressure to provide a residue. The residue was absorbed on HM-N and purified by flash chromatography (Si-SPE, cyclohexane: ethyl acetate, gradient 100:00 to 40:60) to afford the title compound as a yellow oil (2.67 g, 76%). 1H NMR (CDCl3, 400 MHz) 9.05 (s, IH), 8.82 (s, IH), 5.05 (s, 2H), 4.41 (q, J = 7.1 Hz, 2H), 4.24 (q, J=7.1 Hz, 2H), 1.40 (t, J = 7.1 Hz, 3H), 1.28 (t, J = 7.1 Hz, 3H). [00321] Step 4: Ethyl 5-hydroxy-furo|"2,3-d1pyrimidine-6-carboxylate
Figure imgf000081_0002
[00322] To a solution of ethyl 4-ethoxycarbonylmethoxy-pyrimidine-5-carboxylate
(2.12 g, 8.3 mmol) in anhydrous THF (80 ml) at 5°C under an inert atmosphere was added sodium tert-butoxide (1.40 g). The reaction mixture was stirred for 30 minutes at 5°C and a IM solution of hydrochloric acid was added. The mixture was diluted with water and extracted into ethyl acetate. The organic layer was separated and washed with water followed by brine, dried over sodium sulfate, filtered and concentrated under reduced pressure to afford the title compound as a white solid (934 mg, 54%). 1H NMR (CDCl3, 400 MHz) 9.22 (s, IH), 9.14 (s, IH), 4.51 (q, J = 7.3 Hz, 2H), 1.47 (t, J = 7.3 Hz, 3H). [00323] Step 5: Ethyl 5-trifluoromethanesulfonyloxy-furor2,3-dlpyrimidine-6- carboxylate
Figure imgf000081_0003
[00324] To a solution of ethyl 5-hydroxy-furo[2,3-d]pyrimidine-6-carboxylate (1.2 g,
5.8 mmol) and diisopropylethylamine (1.5 ml, 8.7 mmol) in dimethoxyethane (25 ml) was added N-phenyltrifluoromethanesulfonimide (2.3g, 6.4 mmol). The reaction mixture was heated to reflux and stirred for 1 hour, then cooled to room temperature and concentrated under reduced pressure. The residue was dissolved in ethyl acetate then washed with water, saturated aqueous sodium hydrogencarbonate and brine. The organic layer was dried over sodium sulfate, filtered, absorbed on HM-Ν and purified by flash chromatography (Si-SPE, cyclohexane: ethyl actetate, gradient 100:0 to 50:50) to afford the title compound as a colourless oil (1.5 g, 77%). 1H ΝMR (CDCl3, 400 MHz) 9.23 (s, 2H), 4.54 (q, J = 7.2 Hz, 2H), 1.47 (t, J = 7.2 Hz, 3H).
[00325] Step 6: Ethyl 5-(2-fluoro-4-trimethylsilanyl-phenylamino)-furor2,3- dipyrimidine-6-carboxylate
Figure imgf000082_0001
[00326] A degassed solution of ethyl 5-trifluoromethanesulfonyloxy-furo[2,3- d]pyrimidine-6-carboxylate(1.5 g, 4.4 mmol), 2-fluoro-4-trimethylsilanyl-phenylamine mg, 4.8 mmol), Pd2dba3 (202 mg, 0.22 mmol), Xantphos (127 mg, 0.22 mmol) and K3PO4 (1.9 g, 8.8 mmol) in toluene (20 ml) was heated to reflux and stirred for 4 hours under a nitrogen atmosphere. The reaction mixture was cooled to room temperature and filtered through celite, and the filtered cake was washed with ethyl acetate. The organic layer was washed with water then brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The resultant residue was dissolved in dichloromethane, absorbed on HM- Ν and purified by flash chromatography (Si-SPE, ethyl acetate: cyclohexane, gradient 0: 100 to 40:60) to afford the title compound as an oil that crystallized on standing (1.2 g, 75%). LCMS (method B): Rx = 4.39 min, M+H+ = 374.
[00327] Step 7: Ethyl 5-(2-fluoro-4-iodo-phenylamino)-furor2,3-dlpyrimidine-6- carboxylate
Figure imgf000082_0002
To a solution of ethyl 5-(2-fluoro-4-trimethylsilanyl-phenylamino)-furo[2,3- d]pyrimidine-6-carboxylate(1.2 g, 3.2 mmol) in dichloromethane (10 ml) at 5°C was added a solution of iodine monochloride (674 mg, 4.2 mmol) in dichloromethane (5 ml). The reaction mixture was stirred a 50C for 1 hour, before a saturated aqueous solution of sodium thiosulfate was added. The organic layer was separated and washed with water then brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The resultant residue was triturated with hot ethanol, aged overnight at room temperature. The resulting precipitate collected by filtration then washed with cold ethanol before being dried under vacuum to afford the title compound as a white solid (864 mg, 63%). 1H NMR (CDCl3, 400 MHz) 9.08 (s, IH), 8.70 (s, IH), 7.78 (s, IH), 7.57 (dd, J=9.6 Hz, 1.9 Hz, IH), 7.51 (ddd, J=8.4 Hz, 1.7Hz, 1.7Hz, IH), 7.03 (dd, J=8.2Hz, 8.2 Hz, IH), 4.49 (q, J = 7.4 Hz, 2H), 1.46 (t, J = 7.4 Hz, 3H).
[00328] 3-((2-Fluoro-4-iodophenyl)methylamino)furor3,2-clpyridine-2-carboxylic acid ethyl ester
Figure imgf000083_0001
[00329] Sodium hydride (60% dispersion in mineral oil, 45 mg, 1.12 mmol) was added portionwise to a stirred solution of 3-(2-fluoro-4-iodo-phenylamino)-furo[3,2-c]pyridine-2- carboxylic acid ethyl ester (430 mg, 1.0 mmol) and iodomethane (310 μL, 4.98 mmol) in DMF (3 mL) under an inert atmosphere. This mixture was stirred for 3 hours, then quenched with brine and extracted with ethyl acetate (3 x 40 mL). The combined organic extracts were washed with brine, dried (MgSO4), filtered and concentrated in vacuo. The resultant residue was purified using flash chromatography (Si-SPE gradient 40: 100 to 100: 100 ether) to afford the title compound as a yellow solid (57 mg, 13%). LCMS (method B): Rx = 3.26min; M+H+ 440.
[00330] 3-(4-Bromo-2-chloro-phenylamino)-furo[3.2-clpyridine-2-carboxylic acid ethyl ester
Figure imgf000083_0002
[00331] A degassed solution of 3-trifluoromethanesulfonyloxy-furo[3,2-c]pyridine-2- carboxylic acid ethyl ester (300 mg, 0.88 mmol), 4-bromo-2-fluoro aniline (201 mg, 0.97 mmol), Pd2dba3 (40 mg, 0.044 mmol), Xantphos (59 mg, 0.044 mmol) and potassium phosphate tribasic (373 mg, 1.76 mmol) in toluene (5 ml) was heated at reflux under an argon atmosphere for 16 hours. The reaction mixture was filtered and concentrated in vacuo. The resultant residue was purified by flash chromatography (Si-SPE, pentane: diethyl ether, gradient 80:20 to 50:50) to afford the title compound as a yellow solid (177 mg, 51%). LCMS (method B): Rx = 3.76 min, M+H+ = 395 / 397.
[00332] 3-(4-Methyl-2-fluoro-phenylamino)-furor3,2-clpyridine-2-carboxylic acid ethyl ester
Figure imgf000084_0001
[00333] A degassed solution of 3-amino-furo[3,2-c]pyridine-2-carboxylic acid ethyl ester (300 mg, 1.46 mmol), 4-bromo-3-fluorotoluene (277 μl, 2.19 mmol), Pd2dba3 (67 mg, 0.073 mmol), Xantphos (84 mg, 0.15 mmol) and potassium phosphate tribasic (620 mg, 2.92 mmol) in toluene (10 ml) was heated at reflux under an argon atmosphere for 16 hours. The reaction mixture was concentrated in vacuo and the residue was purified by flash chromatography (Si-SPE, pentane: diethyl ether, gradient 100:0 to 75:25) to afford the title compound as a yellow solid (252 mg, 55%). LCMS (method B): Rx = 3.14 min, M+H+ = 315. [00334] 3-(2-Fluoro-4-methylsulfanyl-phenylamino)-furor3,2-c1pyridine-2-carboxylic acid ethyl ester
Figure imgf000084_0002
[00335] Step 1: 4-Bromo-3-fluoro-benzenethiol
Figure imgf000084_0003
[00336] 4-Bromo-3-fluoro-benzenesulfonyl chloride (324 μl, 2.19 mmol) was added dropwise to a solution of triphenylphosphine (1.73 g, 6.58 mmol) in a mixture of dimethylformamide (125 μl) and dichloromethane (5 ml). The solution was stirred at room temperature for 16 hours, then 1 M aqueous hydrochloric acid (5 ml) was added and the layers were separated. The organic layer was concentrated in vacuo and the resultant residue taken up in 1 M aqueous sodium hydroxide (10 ml). The resulting suspension was filtered through celite ® and the filtrate washed with ether (10 ml x 3), then neutralised by addition of 1 M aqueous hydrochloric acid (10 ml). The solution was extracted with ether (10 ml x 3) and the combined organic extracts were dried (Na2SO4) then concentrated in vacuo to afford the title compound as a colourless oil (225 mg, 50%). IH NMR (CDCl3, 300 MHz) 7.47 (IH, dd, J= 8.4, 7.5 Hz), 7.06 (IH, dd, J= 8.9, 2.2 Hz), 6.93 (IH, ddd, J= 8.4, 2.1, 0.7 Hz), 3.54 (IH, br s). [00337] Step 2: l-Bromo-2-fluoro-4-methylsulfanyl -benzene
Figure imgf000085_0001
[00338] A solution of 4-bromo-3-fluoro-benzenethiol (225 mg, 1.09 mmol) in tetrahydrofuran (3 ml) was cooled to O0C. Sodium hydride (60% dispersion in mineral oil, 52 mg, 1.31 mmol) was added and the mixture was stirred for 5 minutes. Iodomethane (78 μl, 1.25 mmol) was then added and the mixture was allowed to return to room temperature with stirring over 20 minutes. Dichloromethane (10 ml) was added and the reaction was quenched with 1 M aqueous hydrochloric acid. The layers were separated and the organic layer was washed with water, dried (MgSO4), then concentrated in vacuo. The residue was purified by flash chromatography (Si-SPE, pentane: diethyl ether, gradient 100:0 to 90:10) to afford the title compound as a bright yellow oil (208 mg, 86%). IH NMR (CDCl3, 400 MHz) 7.43 (IH, dd, J= 8.4, 7.2), 7.00 (IH, dd, J= 9.4, 2.3), 6.91 (IH, ddd, J=8.4, 2.1, 0.7), 2.48 (3 H, s). [00339] Step 3 : 3-(2-Fluoro-4-methylsulfanyl-phenylamino)-furor3,2-c1pyridine-2- carboxylic acid ethyl ester
Figure imgf000085_0002
[00340] A degassed solution of 3-amino-furo[3,2-c]pyridine-2-carboxylic acid ethyl ester (121 mg, 0.59 mmol), l-bromo-2-fluoro-4-methylsulfanyl-benzene (195 mg, 0.88 mmol), Pd2dba3 (27 mg, 0.030 mmol), Xantphos (34 mg, 0.059 mmol) and potassium phosphate tribasic (250 mg, 1.18 mmol) in toluene (3 ml) was heated at reflux under an argon atmosphere for 60 hours. The reaction mixture was filtered and the filtrate concentrated in vacuo. The resultant residue was purified by flash chromatography (Si-SPE, pentane: diethyl ether, gradient 100:0 to 50:50) to afford the title compound as a yellow solid (128 mg, 63%). LCMS (method B): Rx = 3.24 min, M+H+ = 347.
[00341] 7-Chloro-3-(2-fluoro-4-iodo-phenylamino)-furor3,2-c1pyridine-carboxylic acid ethyl ester
Figure imgf000086_0001
[00342] Step 1 : 4,5-Dichloropyridine-3-carbaldehvde
Figure imgf000086_0002
[00343] To a solution of diisopropylamine (10.73 ml, 75.9 mmol) in THF (60 ml) at - 4O0C, was added n-butyllithium (47.45 ml, 75.9 mmol, 1.6M in hexanes) and the solution was stirred for 15 min at -4O0C, before cooling to -7O0C. A solution of 3,4-dichloropyridine (10.7 g, 72.3 mmol) in THF (30 ml) was added dropwise to maintain the temperature below -650C. The reaction was stirred at -7O0C for 2 h before the addition of DMF (6.74 ml, 86.8 mmol). The reaction was then stirred at -4O0C for 1 h and then allowed to warm to -50C before the careful addition of saturated ammonium chloride solution (50 ml) with rapid stirring over 3 min. The mixture was then partitioned between saturated ammonium chloride (150 ml) and dichloromethane (150 ml) and the layers separated. The aqueous layer was extracted with dichloromethane (2 x 100 ml) and the combined organic layers were dried over magnesium sulfate, then concentrated in vacuo. Purification by flash chromatography (Si-SPE, dichloromethane : ethyl acetate gradient 100:0 to 94:6) afforded the title compound as white waxy solid (8.01 g, 63%). [00344] Step 2: 4.5-Dichloropyridine-3-carbaldehyde oxime
Figure imgf000086_0003
[00345] A solution of 4,5-dichloropyridine-3-carbaldehyde (8.01 g, 45.51 mmol) in ethanol (50 ml) was added to a rapidly stirred solution of hydroxylamine hydrochloride (3.48 g, 50.06 mmol) in water (50 ml). The reaction was stirred at r.t. for 45 min then partitioned between ethyl acetate (100 ml) and water (100 ml). The aqueous layer was extracted with ethyl acetate (2 x 50 ml) and the combined organic layers were dried over magnesium sulfate before being concentrated in vacuo to afford the title compound as a white solid (8.3 g, 96%). [00346] Step 3: 4,5-Dichloronicotinonitrile
Figure imgf000087_0001
[00347] To a suspension of 4,5-dichloropyridine-3-carbaldehyde oxime (7.84 g, 41.05 mmol) in dichloromethane (150 ml) was added carbonyl diimidazole (7.99 g , 49.26 mmol). The mixture was then heated to reflux for 1.5 h before cooling then washing with saturated aqueous sodium bicarbonate (70 ml) and water (70 ml). The organic layer was dried over magnesium sulfate and concentrated in vacuo. Purification of the resultant residue by flash chromatography (Si-SPE, cyclohexane : dichloromethane gradient 20:80 to 0: 100) afforded the title compound as a white solid (0.53 g, 72%). LCMS (method B): Rτ = 2.86 min, no ion present. [00348] Step 4: 3-Amino-7-chloro-furor3,2-c"|pyridine-2-carboxylic acid ethyl ester
Figure imgf000087_0002
[00349] To a solution of ethyl glycolate (1.48 ml, 15.7 mmol) in DMF (15 ml) at -1O0C was added sodium hydride (0.63 g, 15.7 mmol, 60% dispersion in oil). The mixture was stirred at this temperature for 35 min then cooled to -4O0C. A solution of 3,4- dichloronicotinonitrile (0.906 g, 5.24 mmol) in DMF (5 ml) was added dropwise before allowing the reaction to warm to -150C for 30 min then -50C for 1 h. The mixture was poured into 10: 1 water/acetic acid (25 ml), diluted with water (25 ml) and extracted with ethyl acetate (2 x 30 ml). The aqueous phase was taken to pH 8 with saturated aqueous sodium bicarbonate then extracted with ethyl acetate (2 x 25 ml). The combined organic layers were dried over magnesium sulfate and concentrated in vacuo. Purification of the resultant residue by flash chromatography (Si-SPE, ethyl acetate : triethylamine 98:2) afforded the title compound as a yellow solid (0.60 g, 48%). LCMS (method B): Rx = 2.79 min, M+H+ = 241, 243. [00350] Step 5: 7-Chloro-3-(2-fluoro-4-trimethylsilanyl-phenylamino)-furor3,2- clpyridine-carboxylic acid ethyl ester
Figure imgf000088_0001
[00351] To a solution of 3-amino-7-chloro-furo[3,2-c]pyridine-2-carboxylic acid ethyl ester (4.16 g, 17.3 mmol) in toluene (100 ml) was added caesium carbonate (11.27 g, 34.6 mmol) and the mixture was degassed (argon / vacuum). To this was added trifluoro- methanesulfonic acid 2-fluoro-4-trimethylsilanyl-phenyl ester (7.1 g, 22.5 mmol), Pd2dba3 (395 mg, 0.432 mmol) and Xantphos (0.5 g, 0.865 mmol) and the vessel was flushed with argon. The reaction mixture was heated to reflux for 19 h, cooled and poured into saturated ammonium chloride (150 ml). The aqueous layer was extracted with ethyl acetate (3 x 60 ml), the combined organic layers were dried over magnesium sulfate and concentrated in vacuo. Purification by flash chromatography (Si-SPE, cyclohexane : dichloromethane gradient 1 :0 to 0: 1) afforded the title compound as a pale yellow solid (5.13 g, 73%). LCMS (method B): Rx = 4.80 min, M+H+ = 407, 409.
[00352] Step 6: 7-Chloro-3-(2-fluoro-4-iodo-phenylamino)-furor3,2-c1pyridine- carboxylic acid ethyl ester
Figure imgf000088_0002
[00353] To a solution of 7-chloro-3-(2-fluoro-4-trimethylsilanyl- phenylamino)furo[3,2c]-pyridine-carboxylic acid ethyl ester (250 mg, 0.615 mmol) in dichloromethane (25 ml) at O0C was added iodine monochloride (1.23 ml, 1.23 mmol, IM solution in dichloromethane) and the solution was stirred at this temperature for 1 hour. A saturated solution of sodium thiosulfate (5 ml) was added and the mixture was poured into saturated sodium thiosulfate (25 ml). The aqueous layer was extracted with dichloromethane (2 x 25 ml), the combined organic layers were washed with brine, dried over magnesium sulfate and concentrated in vacuo. Purification of the resultant residue by flash chromatography (Si-SPE, cyclohexane : dichloromethane gradient 1:0 to 0: 1) afforded the title compound as a yellow waxy solid (0.22 g, 78%). LCMS (method B): Rx = 4.30 min, M+H+ = 461, 463. [00354] 7-Cyano-3-(2-fluoro-4-iodo-phenylamino)-furor3,2-clpyridine-carboxyric acid ethyl ester
Figure imgf000089_0001
[00355] Step 1 : 7-Cvano-3-(2-fluoro-4-trimethylsilanyl-phenylamino)-furor3,2- clpyridine-carboxylic acid ethyl ester
Figure imgf000089_0002
[00356] To a solution of 7-chloro-3-(2-fluoro-4-trimethylsilanyl- phenylamino)furo[3,2c]-pyridine-carboxylic acid ethyl ester (0.64 g, 1.57 mmol) in DMF (15 ml) was added zinc (II) cyanide (0.22 g, 12.63 mmol) and the mixture was degassed (argon/vacuum). Pd2dba3 (72 mg, 0.079 mmol) and 2-dicyclohexylphosphino-2',6'- dimethoxy-l,r-biphenyl (S-Phos, 65 mg, 0.158 mmol) were then added and the vessel was flushed with argon, sealed then heated to 15O0C under microwave irradiation for 30 minutes. The reaction was cooled, the volatiles removed and the residue azeotroped with toluene (3 x 15 ml). Purification of the resultant residue by flash chromatography (Si-SPE, cyclohexane : dichloromethane gradient 1:0 to 0: 1 then 10% ethyl acetate in dichloromethane) afforded the title compound as a pale yellow solid (0.46 g, 74%). LCMS (method B): Rx = 4.52 min, M+H+ = 398.
[00357] Step 2: 7-Cyano-3-(2-fluoro-4-iodo-phenylamino)-furo|"3.2-c]pyridine- carboxylic acid ethyl ester
[00358] To a solution of 7-cyano-3-(2-fluoro-4-trimethylsilanyl- phenylamino)furo[3,2c]-pyridine-carboxylic acid ethyl ester (0.46 g, 1.16 mmol) in dichloromethane (40 ml) at O0C was added iodine monochloride (2.32 ml, 2.32 mmol, IM solution in dichloromethane), and the resultant mixture was stirred at this temperature for 30 minutes. A saturated solution of sodium thiosulfate (5 ml) was added and the mixture poured into saturated sodium thiosulfate (35 ml). The aqueous layer was extracted with dichloromethane (2 x 25 ml) and the combined organic layers were washed with brine, dried over magnesium sulfate then concentrated in vacuo. Purification of the resultant residue by flash chromatography (Si-SPE, dichloromethane : ethyl acetate gradient 10:0 to 10: 1) afforded the title compound as a yellow waxy solid (0.36 g, 69%). LCMS (method B): Rτ = 4.10 min, M+H+ = 452.
[00359] 3-(2-Fluoro-4-triisopropylsilanyloxymethyl-phenylamino)-furo[3.2- cipyridine-2-carboxylic acid ethyl ester
Figure imgf000090_0001
[00360] Step 1 : (4-Bromo-3-fluoro-benzyloxy)-triisopropyl-silane.
Figure imgf000090_0002
[00361] To a solution of (4-bromo-3-fluoro-phenyl)-methanol (410 mg, 2.0 mmol) and imidazole (163 mg, 2.4 mmol) in DMF (10 mL) was added triisopropylsilyl chloride (0.472 mL, 2.2 mmol). The reaction mixture was stirred at room temperature for 18 hours and then partitioned between ethyl acetate and water. The organic layer was isolated, washed with brine, dried (Na2SO4), filtered and concentrated in vacuo. The resultant residue was purified by flash chromatography (Si-SPE, pentane) to provide the title compound as a colourless oil (643 mg, 89%). IH NMR (CDCl3, 400MHz) 7.48 (dd, J=8.1, 7.0 Hz, IH), 7.16 (d, J = 9.7 Hz, IH), 6.99 (d, J=8.7 Hz, IH), 4.78 (s, 2H), 1.04-1.24 (m, 21H).
[00362] Step 2: 3-(2-Fluoro-4-triisopropylsilanyloxymethyl-phenylamino)-furo|"3.2- cipyridine-2-carboxylic acid ethyl ester
[00363] A degassed solution of 3-amino-furo[3,2-c]pyridine-2-carboxylic acid ethyl ester (206 g, 1.0 mmol), (4-bromo-3-fluoro-benzyloxy)-triisopropyl-silane (433 mg, 1.2 mmol), Pd2dba3 (36 mg, 0.039 mmol), Xantphos (46 mg, 0.08 mmol) and K3PO4 (297 mg, 1.4 mmol) in toluene (1 ml) was heated to 1100C then stirred for 4 hours. The reaction mixture was cooled to ambient temperature then diluted with EtOAc and filtered through a pad of celite. The filtrate was concentrated in vacuo to give a black oil. The oil was purified by flash chromatography (Si-SPE, MeOH: DCM, gradient 0: 100 to 5:95) to provide the title compound as a yellow oil (166 mg, 34%). LCMS (method B): Rx = 5.39 min, M+H+ = 487. [00364] 3-(2-Fluoro-4-methoxy-phenylamino)-furor3.2-clpyridine-2-carboxylic acid ethyl ester
Figure imgf000091_0001
[00365] Step 1 : l-Bromo-2-fluoro-4-methoxy-benzene.
Figure imgf000091_0002
[00366] To a solution of 4-bromo-3-fluoro-phenol (500 mg, 2.62 mmol) in anhydrous
THF (10 mL) was added sodium hydride (60% dispersion in mineral oil, 115 mg, 2.88 mmol) portionwise. The reaction mixture was stirred for 20 min before iodomethane (0.500 mL, 8.0 mmol). The resultant mixture was stirred at room temperature for 16 hours before being partitioned between EtOAc and water. The organic layer was separated, washed with a saturated solution of sodium hydrogencarbonate followed by brine, dried (Na2SO4), filtered and concentrated to give the title compound as a pale yellow oil (518 mg, 96%). IH NMR (CDCl3, 400MHz) 7.41 (dd, J=8.8, 8.0 Hz, IH), 6.69 (dd, J = 10.3, 2.8 Hz, IH), 6.61 (ddd, J=8.8, 2.8, 1.0 Hz, IH), 3.79 (s, 3H).
[00367] Step 2: 3-(2-Fluoro-4-methoxy-phenylamino)-furor3,2-c1pyridine-2- carboxylic acid ethyl ester
[00368] A degassed solution of 3-amino-furo[3,2-c]pyridine-2-carboxylic acid ethyl ester (206 g, 1.0 mmol), l-bromo-2-fluoro-4-methoxy-benzene (246 mg, 1.2 mmol), Pd2dba3 (46 mg, 0.050 mmol), Xantphos (58 mg, 0.10 mmol) and K3PO4 (254 mg, 1.2 mmol) in toluene (5 ml) was heated to 1100C then stirred for 18 hours. The reaction mixture was cooled to ambient temperature then diluted with EtOAc and filtered through a pad of celite. The filtrate was concentrated in vacuo to give a black oil. The oil was purified by flash chromatography (Si-SPE, MeOH: DCM, gradient 0: 100 to 10:90) to provide the title compound as a yellow oil (130 mg, 39%). LCMS (method B): Rx = 2.93 min, M+H+ = 331. [00369] Ethyl 3-(4-Bromo-2,5-difluorophenylamino)furo[3,2-clpyridine-2-carboxylate
Figure imgf000091_0003
[00370] A degassed solution of ethyl 3-(trifluoromethanesulfonyloxy)furo [3,2- c]pyridine-2-carboxylate (678 mg, 2.00 mmol), 4-bromo-2,5-difluoroaniline (670 mg, 3.22 mmol), Pd2dba3 (147 mg, 0.160 mmol), Xantphos (97.0 mg, 0.168 mmol) and finely powdered K3PO4 (793 mg, 3.74 mmol) in toluene (7.5 ml) was heated in a sealed tube at 1050C overnight. The reaction mixture was cooled to ambient temperature then diluted with ethyl acetate (15 ml) and filtered through a plug of silica gel (15 ml packed in ethyl ether). After washing the filter cake with more ethyl acetate (20 ml), the filtrate was dried over magnesium sulfate and concentrated in vacuo to give a brown oil. The residue was purified by flash chromatography (silica gel, using using 5:3:2 hexane-methylene chloride-ethyl ether) to afford the title compound as a tan solid (329 mg, 41%). [00371] 3-(2-Fluoro-4-iodo-phenoxy)-furor3,2-clpyridine-2-carboxylic acid ethyl ester
[00372] Step 1 : 3-(2-Fluoro-4-nitro-phenoxy)-furor3.2-c1pyridine-2-carboxylic acid ethyl ester
Figure imgf000092_0002
[00373] Ethyl 3-hydroxy-furo[3,2-c]pyridine-2-carboxylate (2.70 g, 13.0 mmol), followed by 3,4-difluoro nitrobenzene (2.89 mL, 26.1 mmol), and 18-crown-6 (3.45 g, 13.0 mmol) were added to a suspension of potassium hydride (1.10 g, 27.4 mmol) in DMF (30 mL) at room temperature. The reaction mixture was heated to 100 0C for 2h, then cooled to room temperature and poured into a water / brine mixture. The aqueous layer was extracted 3 times with EtOAc, then the combined organics were washed once with brine, dried over Na2SO4 and concentrated. The residue was purified by silica chromatography (30 - 80% EtOAc: Hex) to afford the title compound (442 mg, 10% yield) as a yellow syrup. LCMS (method C): Rx = 2.07 min, M+H+ = 347 . 1H NMR (CDCl3, 400MHz) 8.83 (d, J= 1.2 Hz, IH), 8.72 (d, J= 6.0 Hz, IH), 8.17 (dd, J= 10.0, 2.4 Hz, IH), 8.03 (ddd, J= 9.2, 2.8, 1.6 Hz, IH), 7.59 (dd, J= 5.6, 0.8 Hz, IH), 7.08 (dd, J= 9.2, 8.0 Hz, IH), 4.36 (q, J= 7.2 Hz, 2H), 1.28 (t, J= 7.2 Hz, 3H).
[00374] Step 2: 3-(4-Amino-2-fluoro-phenoxy)-furor3,2-c1pyridine-2-carboxylic acid ethyl ester
Figure imgf000093_0001
[00375] Fe powder (299 mg, 5.36 mmol) was added to a solution of 3-(2-Fluoro-4- nitro-phenoxy)-furo[3,2-c]pyridine-2-carboxylic acid ethyl ester (460 mg, 1.3 mmol) in ethanol (8 mL) and 2N aqueous HCl (8 mL). The reaction mixture was heated to 50 0C, then cooled to room temperature. The unreacted iron was removed with a magnet, then the reaction mixture was concentrated in vacuo. 10 mL each of water and ethanol were added, followed by solid sodium bicarbonate (1.5g). 3.2 g of silica gel was added and the volatiles were removed in vacuo. The residue was purified by silica chromatography (40 - 80% EtOAc: Hex) to afford the title compound (130 mg, 31% yield) as an off-white foam. LCMS (method C): Rx = 1.35 min, M+H+ = 317. 1H NMR (CDCl3, 400MHz) 8.54 (d, J= 3.6 Hz, IH), 8.16 (d, J= 0.8 Hz, IH), 7.45 (dd, J= 5.6, 0.8 Hz, IH), 7.09 (t, J= 8.8 Hz, IH), 6.53 (dd, J= 12.4, 2.8 Hz, IH), 6.44 (ddd, J= 8.8, 2.8, 1.6 Hz, IH), 4.47 (q, J= 7.2 Hz, 2H), 3.81 (br, 2H), 1.42 (t, J= 7.2 Hz, 3H).
[00376] Step 3: 3-(2-Fluoro-4-iodo-phenoxy)-furor3,2-clpyridine-2-carboxylic acid ethyl ester
[00377] Sodium nitrite (1.18 mL of a 0.382 M aqueous solution) was added dropwise to a suspension of 3-(4-Amino-2-fluoro-phenoxy)-furo[3,2-c]pyridine-2-carboxylic acid ethyl ester (130 mg, 0.41 mmol) in a 2 M aqueous solution of HCl (3.5 mL) at 0 0C. The reaction mixture was stirred for 45 minutes at 0 0C, and then sodium iodide (1.18 mL of a 1.39 M aqueous solution, 1.64 mmol) was added. The reaction mixture was stirred overnight at 0 0C - room temperature. Sodium hydroxide (7 mL of IN aqueous solution) and Na2S2θ3 (5 mL of saturated aqueous solution) were added, and the aqueous layer was extracted three times with CH2Cl2. The combined organics were dried over Na2SO4 , filtered and concentrated. The residue was purified by silica chromatography (30 - 70% EtOAc:Hex) to afford the title compound (60 mg, 30% yield) as a white solid. LCMS (method C): Rτ = 2.29 min, M+H+ = 428. 1H NMR (CDCl3, 400MHz) 8.63 (d, J= 6.4 Hz, IH), 8.53 (d, J= 1.2 Hz, IH), 7.58 (dd, J= 9.6, 2.0 Hz, IH), 7.52 (dd, J= 6.0, 1.2 Hz, IH), 7.37 (dt, J= 8.8, 1.6 Hz, IH), 6.86 (t, J = 8.4 Hz, IH), 4.41 (q, J= 7.2 Hz, 2H), 1.34 (t, J= 7.2 Hz, 3H).
[00378] 3-(2-Fluoro-44odophenylamino)-7-phenylfuro|"3,2-c]pyridine-2-carboxyric acid ethyl ester
Figure imgf000094_0001
[00379] Step 1 : 4,5-Dibromonicotinic acid
Figure imgf000094_0002
[00380] 5-Bromonicotinic acid (25.25 g, 125 mmol) was stirred as a solution in dry tetrahydrofuran (500 ml) under nitrogen and cooled to -7O0C. The resultant mixture was treated dropwise over 1 hour with lithium diisopropylamide (1.8 M, 144 ml, 260 mmol). After the addition was complete, the solution was stirred for 2.5 hours at - 550C then cooled to -7O0C and treated portionwise over 30 minutes with 1,2-dibromotetrachloroethane (50 g, 154.5 mmol). After stirring for 30 minutes the mixture was allowed to warm to -2O0C over 2 hours before the cautious addition water (150 ml). The organic solvent was then removed in vacuo and the residue diluted with water (500 ml), then washed with ethyl acetate before acidifying the aqueous layer to pH 3.00 with c. HCl. The precipitated product was collected by filtration and dried at 6O0C in vacuo to give the title compound (14.2g). The filtrate was extracted with ethyl acetate, the extract washed with water, dried (MgSO4), filtered and concentrated in vacuo to give further title compound (18.6g, total yield 32.8 g, 93%). 1H NMR (DMSO-d6, 400MHz) 8.92 (s, IH), 8.73 (s, IH). [00381] Step 2: 4.5-Dibromonicotinic acid ethyl ester
Figure imgf000094_0003
[00382] 4,5-Dibromonicotinic acid (32.8 g, 116.7 mmol) was stirred as a suspension in acetonitrile (550 ml) at room temperature and treated portionwise with 1,1' - carbonyldiimidazole (29.87 g, 180 mmol) over 10 minutes. The resulting mixture was stirred for 3 hours at room temperature. After this time ethanol (78 ml) was added and stirring continued for a further 48 hours. The solution was then filtered and the filtrate evaporated in vacuo to give a light brown oil. The oil was dissolved in ethyl acetate and the solution was washed water followed by brine then dried over magnesium sulfate, filtered and evaporated in vacuo to give a brown oil. The oil was purified by flash chromatography (SiC^ dichloromethane eluent) to give the title compound (20.6 g 57 %) IH NMR (CDCl3, 400 MHz) 8.80 (s, IH), 8.75 (s, IH), 4.45 (q, 2H J = 7.0 Hz), 1.39 (t, 3H J= 7.0 Hz)
[00383] Step 3: 7-Bromo-3-hvdroxyfuror3,2-clpyridine-2-carboxylic acid ethyl ester
Figure imgf000095_0001
[00384] A solution ethyl glycolate (6.30 ml, 66.5 mmol) in dry DMF (50 ml) was added dropwise to a stirred suspension of sodium hydride (8.00 g, 60% dispersion, 200 mmol) in dry DMF (80 ml) whilst cooling to maintain the temperature below 1O0C. After the addition, the mixture was stirred for 30 minutes before the dropwise addition of 4,5- dibromonicotinic acid ethyl ester (20.60 g, 66.5 mmol) as a solution in dry DMF (50ml) whilst again maintaining the temperature below 1O0C. The resulting dark red/brown solution was allowed to warm to room temperature slowly over 1.5 hours before quenching and acidifying to pH 3.00 with aqueous IM HCl. The resulting solid precipitate was collected by filtration, the residue was washed with water then cold acetone and dried in vacuo at 450C to give the title compound (11.98 g 63%). IH NMR (DMSO-d6, 400MHz) 9.13 (s,lH), 8.76 (s, IH), 4.35 (q, 2H J= 7.3 Hz), 1.33 (t, 3H J= 7.3 Hz). LCMS (method B): Rx = 2.82 min, M+H+ = 286, 288. [00385] Step 4: 7-Bromo-3-trifluoromethanesulfonyloxyfuror3 ,2-clpyridine-2- carboxylic acid ethyl ester
Figure imgf000095_0002
[00386] Trifluoromethanesulfonic anhydride (8.32 ml, 49.66 mmol) in dry DCM (70 ml) was added dropwise to a stirred solution of 7-bromo-3-hydroxyfuro[3,2-c]pyridine-2- carboxylic acid ethyl ester (12.80 g, 44.7 mmol) and pyridine (10.88 ml, 128 mmol) in dry DCM (400 ml) at 5-1O0C. The resulting mixture was stirred for 1.5 hours at 5-1O0C then allowed to warm to room temperature slowly over 3 hours before being left to stand for 16 hours. The mixture was diluted with DCM, washed with IM aqueous HCl, water, saturated aqueous NaHCO3 and brine, before being dried over magnesium sulfate, filtered and concentrated in vacuo to give light brown oil. The oil was purified by flash chromatography (Siθ2, dichloromethane) to give the title compound as a yellow solid (11.84 g 63%). IH NMR (CDCl3, 400 MHz) 8.96 (s, IH), 8.83 (s, IH), 4.55 (q, 2H J = 7.2 Hz), 1.47 (t, 3H J= 7.2 Hz).
[00387] Step 5: 7-Bromo-3-(2-fluoro-4-trimethylsilanylphenylamino)furor3,2- cipyridine-2-carboxylic acid ethyl ester
Figure imgf000096_0001
[00388] 7-Bromo-3-trifluoromethanesulfonyloxyfuro[3,2-c]pyridine-2-carboxylic acid ethyl ester (11.84 g, 28.3 mmol) was stirred in dry toluene (160 ml) with Pd2(dba)3 (1.0 g, 1.20 mmol), Xantphos (0.572 g, 1.0 mmol) and potassium phosphate tribasic (11.25 g, 53.75 mmol). The mixture was de-gassed before the addition of a solution 2-fluoro-4- trimethylsilanylphenylamine (5.38 g, 29.54 mmol) in dry toluene (10 ml). The mixture was de-gassed again before heating at 1150C for 4 hours. The reaction mixture was partitioned between ethyl acetate and water then filtered and the layers separated. The organic layer was washed with water then brine, dried over magnesium sulfate, filtered and evaporated in vacuo to give a brown solid. The solid was purified by flash chromatography (SiO2, 30% cyclohexane in DCM ) to give the title compound as a pale yellow solid (7.1 g, 55%). IH NMR (CDCl3, 400 MHz) 8.66 (s, IH), 8.50 (s, IH), 7.73 (s, IH), 7.27 (m, 3H), 4.50 (q, 2H J = 7.0 Hz), 1.47 (t, 3H J= 7.0 Hz), 0.29 (s, 9H). LCMS (method B): Rx = 4.81 min, M+H+ = 451, 453. [00389] Step 6: 3-(2-Fluoro-4-trimethylsilanylphenylamino)-7-phenylfuror3,2- clpyridine-2-carboxylic acid ethyl ester
Figure imgf000096_0002
[00390] 7-Bromo-3-(2-fluoro-4-trimethylsilanylphenylamino)furo[3,2-c]pyridine-2- carboxylic acid ethyl ester (100 mg, 0.22 mmol) was stirred as a suspension at room temperature in ethanol (2 ml) with phenyl boronic acid (30 mg, 0.242 mmol) under argon. After stirring for 20 minutes Pd(OAc)2 (2 mg, 0.66 μmol), triphenyl phosphine (0.5 mg, 0.002 mmol) and 2M aqueous Na2CO3 (130 μl, 0.264 mmol) were added and the reaction mixture de-gassed then heated at reflux under argon for 3 hours. The reaction mixture was cooled to room temperature and diluted with water then extracted with ethyl acetate. The organic layer was washed with water then brine and then dried over magnesium sulfate, filtered and evaporated in vacuo to give a yellow solid. This solid was purified by flash chromatography (SiO2 30% cyclohexane in DCM) to give the title compound as a pale yellow solid (31 mg, 31%). IH NMR (CDCl3 , 400 MHz) 8.73 (s, IH), 8.61 (s, IH), 7.87 (d, 2H J = 7.80 Hz), 7.72 (s,lH), 7.59-7.44 (m, 3H), 7.35 - 7.25 (m, 3H), 4.33 (q, 2H J = 7.0 Hz) 1.43 (t, 3H J= 7.0 Hz) 0.30 (s, 9H). LCMS (method B): Rx = 4.91 min, M+H+ = 449. [00391] Step 7: 3-(2-Fluoro-4-iodophenylamino)-7-phenylfuro|"3,2-c]pyridine-2- carboxylic acid ethyl ester
[00392] 3-(2-Fluoro-4-trimethylsilanylphenylamino)-7-phenylfuro[3,2-c]pyridine-2- carboxylic acid ethyl ester (30 mg, 0.067 mmol) was stirred as a solution in DCM (2 ml) at 0- 50C and treated dropwise with IM ICl in DCM (130 μl, 0.13 mmol). The resulting mixture was stirred at 0-50C for 2 hours before the addition IM aqueous Na2S2θ3 (1 ml). The layers were separated and the organic layer was washed with water, followed by brine, dried over magnesium sulfate, filtered and evaporated in vacuo to give the title compound (quantitative). IH NMR (CDCl3, 400MHz) 8.75 (s, IH), 8.58 (s, IH), 7.87 (d, 2H J = 7.80 Hz), 7.67 (s, IH), 7.59-7.44 (m, 5H), 7.05 (t, IH J = 8.50 Hz), 4.47 (q, 2H J = 7.10 Hz), 1.43 (t, 3H J= 7.10 Hz). LCMS (method B): Rx = 4.40 min, M+H+ = 503. [00393] 3-(2-Fluoro-4-iodophenylamino)-7-methylfuror3,2-c1pyridine-2-carboxylic acid ethyl ester
Figure imgf000097_0001
[00394] Step 1 : 3-(2-Fluoro-4-trimethylsilanylphenylamino)-7-methylfuro[3,2- clpyridine-2-carboxylic acid ethyl ester
Figure imgf000097_0002
[00395] 7-Bromo-3-(2-fluoro-4-trimethylsilanylphenylamino)furo[3,2-c]pyridine-2- carboxylic acid ethyl ester (1.0 g, 2.2 mmol) was stirred with potassium carbonate (456.5 mg, 3.3 mmol), tetrakistriphenylphosphine palladium(O) (255 mg, 0.22 mmol) and trimethylboroxine (305 μl, 2.2 mmol) in dry 1,4-dioxane (5 ml). The reaction mixture was de-gassed before heating at HO0C under argon for 6 hours then cooled to room temperature and left to stand for 16 hours. The reaction mixture was diluted with dichloromethane and water. The organic layer was separated, washed with water flowed by brine, dried over magnesium sulfate, then filtered and evaporated in vacuo to give a residue. The crude residue was purified by flash chromatography (SiO2, 30% cyclohexane in DCM then 1% methanol in DCM) to give the title compound as a pale yellow solid (710 mg, 83%) IH NMR (CDCl3 , 400 MHz) 8.50 (s, IH), 8.38 (s, IH), 7.70 (s, IH), 7.32-7.22 (m, 3H), 4.48 (q, 2H J = 7.0 Hz), 2.54 (s, 3H ), 1.46 (t, 3H J = 7.0 Hz), 0.29 (s 9H). LCMS (method B): Rx = 4.12 min, M+H+ = 387.
[00396] Step 2: 3-(2-Fluoro-4-iodophenylamino)-7-methylfuro|"3.2-c]pyridine-2- carboxylic acid ethyl ester
[00397] 3-(2-Fluoro-4-trimethylsilanylphenylamino)-7-methylfuro[3,2-c]pyridine-2- carboxylic acid ethyl ester (710 mg, 1.84 mmol) was stirred as a solution in DCM (25 ml) at 0-50C and treated drop wise with IM ICl in DCM (3.5 ml, 3.5 mmol). The resulting mixture was stirred at 0-50C for 2 hours before the addition IM aqueous Na2S2O3 (12 mL). The layers were separated and the organic layer was washed with water, brine, then dried over magnesium sulfate, filtered and evaporated in vacuo to give a residue. The crude residue was purified by flash chromatography (SiO2, gradient 0-1% MeOH in DCM) to give the title compound as a pale yellow solid (448 mg, 55%). IH NMR (CDCl3 , 400 MHz) 8.47 (s, IH), 8.39 (s, IH), 7.65 (s, IH), 7.52 (dd, IH J = 9.8, 1.9 Hz), 7.44 (dt, IH J = 8.4, 1.3 Hz), 7.00 (t, IH J = 8.5 Hz), 4.48 (q, 2H J = 7.0 Hz), 2.53 (s, 3H ), 1.46 (t, 3H J = 7.0 Hz). LCMS (method B): Rx = 3.39 min, M+H+ = 441.
[00398] 2-((R)-2.2-Dimethyl-ri.31dioxolan-4-ylmethoxycarbamoyl)-3-(2-fluoro-4- iodo-phenylamino)-furor3,2-clpyridine-7-carboxylic acid ethyl ester
Figure imgf000098_0001
[00399] Step 1 : 3-Hydroxy-furor3.2-clpyridine-2.7-dicarboxylic acid 2-benzyl ester 7- ethyl ester
Figure imgf000099_0001
[00400] To a solution of 4-chloro-pyridine-3,5-dicarboxylic acid diethyl ester (250 mg,
0.971 mmol) and benzyl glycolate (145 μl, 1.019 mmol) in DMF (5 ml) at O0C, was added sodium hydride (97 mg, 2.43 mmol, 60% dispersion in mineral oil). The mixture was allowed to warm to room temperature and stirred for 3 hours then quenched by the addition of acetic acid (1 ml). The mixture was then concentrated in vacuo and the resulting residue was triturated in water and filtered. The resulting solid was recrystallised from methanol/water to afford the title compound as a pale yellow solid (120 mg, 36%). LCMS (method B): Rx = 3.29 min, M+H+ = 342.
[00401] Step 2: 3-Trifluoromethanesulfonyloxy-furor3.2-clpyridine-2.7-dicarboxylic acid 2-benzyl ester 7-ethyl ester
Figure imgf000099_0002
[00402] To a solution of 3-hydroxy-furo[3,2-c]pyridine-2,7-dicarboxylic acid 2-benzyl ester 7-ethyl ester (120 mg, 0.352 mmol) and pyridine (85 μl, 1.056 mmol) in dichloromethane (1.5 ml) at O0C was added trifluoromethanesulfonic anhydride (63 μl, 0.37 mmol) dropwise. The reaction was stirred at room temperature for 90 minutes then partitioned between dichloromethane (30 ml) and 0.1M HCl (10 ml). The organic layer was isolated and washed with saturated sodium bicarbonate (10 ml) then brine (10 ml). The isolated organic layer was dried over magnesium sulfate before being concentrated in vacuo to afford the title compound as a colourless oil (88 mg, 53%). 1H NMR (CDCl3, 400 MHz) 9.27 (IH, s), 9.17 (IH, s), 7.48 (2H, m), 7.38 (3H, m), 5.48 (2H, s), 4.52 (2H, q, J= 7.2 Hz), 1.43 (3H, t, J = 7.2 Hz).
[00403] Step 3 : 3-(2-Fluoro-4-trimethylsilanyl-phenylamino)-furor3,2-c1pyridine-2,7- dicarboxylic acid 2-benzyl ester 7-ethyl ester
Figure imgf000100_0001
[00404] To a solution of 3-trifluoromethanesulfonyloxy-furo[3,2-c]pyridine-2,7- dicarboxylic acid 2-benzyl ester 7-ethyl ester (88 mg, 0.186 mmol) and 2-fluoro-4- trimethylsilanyl-phenylamine (41 mg 0.223 mmol) in toluene (1.5 ml) was added potassium phosphate (55 mg, 0.26 mmol) before the mixture was degassed. Pd2dba3 (8.5 mg, 0.0093 mmol) and Xantphos (11 mg, 0.0186 mmol) were added to this mixture and the vessel was flushed with argon. The reaction mixture was heated to reflux for 1.5 hours, cooled and filtered through Celite ® washing with ethyl acetate. The filtrate was washed with saturated sodium bicarbonate (10 ml), then dried over magnesium sulfate and concentrated in vacuo. Purification of the resultant residue by flash chromatography (Si-SPE, cyclohexane : t-butyl methyl ether gradient 1 :0 to 3: 1) afforded the title compound as a pale yellow solid (48 mg, 51%). LCMS (method B): Rx = 4.89 min, M+H+ = 507.
[00405] Step 4: 3-(2-Fluoro-4-trimethylsilanyl-phenylamino)-furor3,2-c"|pyridine-2J- dicarboxylic acid 7-ethyl ester
Figure imgf000100_0002
[00406] To a solution of 3-(2-fluoro-4-trimethylsilanyl-phenylamino)-furo[3,2- c]pyridine-2,7dicarboxylic acid 2-benzyl ester 7-ethyl ester (48 mg, 0.0949 mmol) in ethyl acetate (2 ml) under nitrogen, was added palladium on carbon (12 mg, 10% palladium on activated charcoal). The suspension was stirred at room temperature for 2 hours under an atmosphere of hydrogen. The reaction mixture was filtered through Celite ®, washing with ethyl acetate, and the filtrate was concentrated in vacuo to afford the title product as a colourless oil (34 mg, 86%). LCMS (method B): Rx = 4.31 min, M+H+ = 417, [M-H]" = 415. [00407] Step 5: 2-((R)-2,2-Dimethyl-rL31dioxolan-4-ylmethoxycarbamoyl)-3-(2- fluoro-4-iodo-phenylamino)-furor3,2-c1pyridine-7-carboxylic acid ethyl ester [00408] To a solution of 2-((R)-2,2-dimethyl-[l,3]dioxolan-4-ylmethoxycarbamoyl)-
3-(2-fluoro-4-trimethylsilanyl-phenylamino)-furo[3,2-c]pyridine-7-carboxylic acid ethyl ester (37 mg, 0.068 mmol) in dichloromethane (2 ml) at -50C was added iodine monochloride (136 μl, 0.136 mmol, IM solution in dichloromethane) and the solution was stirred at this temperature for 1 hour. A saturated solution of sodium thiosulfate (5 ml) was added and the mixture was poured into saturated sodium thiosulfate (15 ml). The aqueous layer was isolated and extracted with dichloromethane (2 x 25 ml), before the combined organic layers were washed with brine, dried over magnesium sulfate and concentrated in vacuo. Purification of the resultant residue by flash chromatography (Si-SPE, dichloromethane : ethyl acetate gradient 1:0 to 0: 1 then 15% methanol in dichloromethane) afforded the title compound as a yellow waxy solid (29 mg, 71%). LCMS (method B): Rx = 3.92 min, M+H+ = 600. [00409] 2-Dimethylcarbamoyl-3-(2-fluoro-4-trimethylsilanyl-phenylamino)-furor3,2- cipyridine-7-carboxylic acid ethyl ester
Figure imgf000101_0001
[00410] Step 1 : 2-Dimethylcarbamoyl-3-hydroxy-furo[3.2-c]pyridine-7-carboxyric acid ethyl ester
Figure imgf000101_0002
[00411] To a solution of 4-chloro-pyridine-3,5-dicarboxylic acid diethyl ester (430 mg,
1.67 mmol) and 2-hydroxy-N,N-dimethyl-acetamide (189 mg, 1.84 mmol) in DMF (7 ml) at O0C was added sodium hydride (200 mg, 5.01 mmol, 60% dispersion in mineral oil). The resultant mixture was allowed to warm to room temperature and stirred for 2.5 hours. The reaction was quenched by the addition of acetic acid (1 ml). The mixture was then concentrated in vacuo and the resulting residue was triturated in water and filtered to afford the title compound as a pale yellow solid (200 mg, 43%). LCMS (method B): RT = 2.73 min, M+H+ = 279.
[00412] Step 2: 2-Dimethylcarbamoyl-3-trifluoromethanesulfonyloxy-furor3,2- cipyridine-7-carboxylic acid ethyl ester
Figure imgf000102_0001
To a solution of 2-dimethylcarbamoyl-3-hydroxy-furo[3,2-c]pyridine-7-carboxylic acid ethyl ester (440 mg, 1.58 mmol) and pyridine (0.38 ml, 4.74 mmol) in dichloromethane (7 ml) at O0C was added trifluoromethanesulfonic anhydride (0.29 ml, 1.74 mmol) dropwise. The reaction was stirred at room temperature for 120 minutes then partitioned between dichloromethane (50 ml) and 0. IM HCl (20 ml). The organic layer was washed with saturated sodium bicarbonate (20 ml) then brine (20 ml). The combined organic layers were dried over magnesium sulfate before being concentrated in vacuo to afford the title compound as a colourless oil (144 mg, 22%). LCMS (method B): Rx = 3.39 min, M+H+ = 411.
[00413] Step 3 : 2-Dimethylcarbamoyl-3-(2-fluoro-4-trimethylsilanyl-phenylamino)- furo|"3.2-c]pyridine-7-carboxyric acid ethyl ester
[00414] To a solution of 2-dimethylcarbamoyl-3-trifluoromethanesulfonyloxy- furo[3,2-c]pyridine-7-carboxylic acid ethyl ester (144 mg, 0.351 mmol) and 2-fluoro-4- trimethylsilanyl-phenylamine (90 mg 0.492 mmol) in toluene (3 ml) was added potassium phosphate (149 mg, 0.70 mmol) before the mixture was degassed. Pd2dba3 (16.1 mg, 0.0176 mmol) and Xantphos (20 mg, 0.035 mmol) were added to the reaction mixture and the vessel was flushed with argon. The reaction mixture was then heated to reflux for 3 hours, cooled and filtered through Hyflo, washing with ethyl acetate. The filtrate was washed with saturated sodium bicarbonate (30 ml), the organic layer dried over magnesium sulfate and concentrated in vacuo. Purification of the resultant residue by flash chromatography (Si-SPE, cyclohexane : t-butyl methyl ether gradient 3: 1 to 1: 1) afforded the title compound as a pale yellow solid (84 mg, 54%). LCMS (method B): Rx = 4.58 min, M+H+ = 444.
[00415] 7-Fluoro-3-(2-fluoro-4-iodo-phenylamino)-furor3,2-c1pyridine-2-carboxylic acid ethyl ester
Figure imgf000102_0002
[00416] Step 1 : 4-Chloro-5-fluoro-nicotinic acid ethyl ester
Figure imgf000103_0001
[00417] A suspension of 4-chloro-5-fluoro-nicotinic acid (0.36 g, 2.06 mmol) in thionyl chloride (3 ml) was heated at 8O0C for 2 hours until most of the solid had dissolved. The reaction mixture was concentrated in vacuo and the residue azeotroped with toluene (2 x 20 ml). The resultant residue was dissolved in ethanol (5 ml) and diisopropylethylamine (1.76 ml, 10.31 mmol) and the reaction mixture was stirred at room temperature for 18 hours. The reaction mixture was concentrated in vacuo, diluted with ethyl acetate and washed with 0. IM HCl then saturated sodium bicarbonate and brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. Purification of the resultant residue by flash chromatography (Si-SPE, dichloromethane : ethyl acetate gradient 1:0 to 92:8) afforded the title compound as a colourless oil (415 mg, 99%). 1H NMR (CDCl3, 400 MHz) 1.43 (3H, t, J= 7.1 Hz), 4.46 (2H, q, J= 7.1 Hz), 8.60 (IH, d, J = 0.8 Hz), 8.86 (IH, s). [00418] Step 2: 7-Fluoro-3-hvdroxy-furor3,2-c"|pyridine-2-carboxyric acid ethyl ester
Figure imgf000103_0002
[00419] To a solution of 4-chloro-5-fluoro-nicotinic acid ethyl ester (400 mg, 1.975 mmol) and ethyl glycolate (196 μl, 2.074 mmol) in DMF (10 ml) at O0C, was added sodium hydride (158 mg, 3.95 mmol, 60% dispersion in mineral oil) and the mixture was allowed to warm to room temperature then stirred for 2 hours. The reaction was quenched by the addition of acetic acid (1.5 ml). The mixture was then concentrated in vacuo and the resulting residue was triturated in water to afford the title compound as a pale yellow solid (460 mg, quantitative). LCMS (method B): Rx = 2.59 min, M+H+ = 226. [00420] Step 3: 7-Fluoro-3-trifluoromethanesulfonyloxy-furo|"3.2-c]pyridine-2- carboxylic acid ethyl ester
F
Figure imgf000103_0003
[00421] To a solution of 7-fluoro-3-hydroxy-furo[3,2-c]pyridine-2-carboxylic acid ethyl ester (460 mg, 1.97 mmol) and pyridine (0.48 ml, 5.91 mmol) in dichloromethane (10 ml) at O0C was added trifluoromethanesulfonic anhydride (612 mg, 2.17 mmol) dropwise. The reaction was stirred at room temperature for 90 minutes then partitioned between dichloromethane (50 ml) and 0. IM HCl (20 ml). The organic layer was isolated and washed with saturated sodium bicarbonate (20 ml) then brine (20 ml). The combined organic layers were dried over magnesium sulfate before being concentrated in vacuo. Purification of the resultant residue by flash chromatography (Si-SPE, dichloromethane) afforded the title compound as a colourless oil (470 mg, 67%). LCMS (method B): Rτ = 3.76 min, M+H+ = 358.
[00422] Step 4: 7-Fluoro-3-(2-fluoro-4-trimethylsilanyl-phenylamino)-furor3,2- clpyridine-2-carboxylic acid ethyl ester
Figure imgf000104_0001
[00423] To a solution of 7-fluoro-3-trifluoromethanesulfonyloxy-furo[3,2-c]pyridine-
2-carboxylic acid ethyl ester (470 mg, 1.32 mmol) and 2-fluoro-4-trimethylsilanyl- phenylamine (337 mg, 1.84 mmol) in toluene (15 ml) was added potassium phosphate (558 mg, 2.63 mmol) and the mixture was degassed. Pd2dba3 (60.5 mg, 0.066 mmol) and Xantphos (76.5 mg, 0.132 mmol) were added to the reaction mixture and the vessel was flushed with argon. The reaction mixture was heated to reflux for 4 hours, cooled and filtered through Hyflo, washing with ethyl acetate. The filtrate was washed with saturated sodium bicarbonate, the organic layer dried over magnesium sulfate and concentrated in vacuo. Purification of the resultant residue by flash chromatography (Si-SPE, dichloromethane : ethyl acetate gradient 1 :0 to 9: 1) afforded the title compound as a pale yellow solid (490 mg, 95%). LCMS (method B): Rx = 4.70 min, M+H+ = 391.
[00424] Step 5: 7-Fluoro-3-(2-fluoro-4-iodo-phenylamino)-furo|"3.2-c]pyridine-2- carboxylic acid ethyl ester
[00425] To a solution of 7-fluoro-3-(2-fluoro-4-trimethylsilanyl-phenylamino)- furo[3,2-c]pyridine-2-carboxylic acid ethyl ester (490 mg, 1.256 mmol) in dichloromethane (8 ml) at -1O0C was added iodine monochloride (2.51 ml, 2.51 mmol, IM solution in dichloromethane) and the solution was stirred between -1O0C and O0C for 2 h. A saturated solution of sodium thiosulfate (5 ml) was added and the mixture was poured into saturated sodium thiosulfate (15 ml). The aqueous layer was isolated then extracted with dichloromethane (3 x 25 ml) before the combined organic layers were washed with brine, dried over magnesium sulfate and concentrated in vacuo. Purification of the resultant residue by flash chromatography (Si-SPE, cyclohexane : ethyl acetate gradient 1 :0 to 3: 1 then dichloromethane) gave crude material. The crude material was triturated in cyclohexane to afford the title compound as a yellow waxy solid (250 mg, 45%). LCMS (method B): RT = 4.13 min, M+H+ = 445.
[00426] 7-Fluoro-3-(2-fluoro-4-iodo-phenylamino)-furor3,2-c1pyridine-carboxylic acid ethyl ester
Figure imgf000105_0001
[00427] Step 1: 4-Chloro-5-fluoro-pyridine-3-carbaldehyde oxime
Figure imgf000105_0002
[00428] To a cooled (-780C) solution of 3-fluoro-4-chloro-pyridine (11.0 g, 84 mmol) in THF under nitrogen was added lithium diisopropylamide (1.8 M solution, 47 mL, 84 mmol) dropwise and the resultant solution stirred at -70 to -8O0C for 18 hours. DMF (7.68 g, 1.25 eq.) was added dropwise and stirring continued at -780C for 30 minutes before adding the reaction mixture to ice/2M HCl. The solution was extracted with diethyl ether and the organic layer back-extracted with 2M HCl, the two aqueous solutions held separately. The aqueous extracts were each treated with hydroxylamine hydrochloride (8.76 g, 126 mmol) and adjusted to pH 5 with potassium carbonate. After stirring for 1 hour the mixtures were extracted with ethyl acetate (x 2), the combined organic extracts dried (Na2SO4), filtered and concentrated in vacuo to afford the title compound as a tan solid (11.07 g, 76%). LCMS (method B): Rx = 2.49 min, M+H+ 175.
[00429] Step 2: 4-Chloro-5-fluoronicotinonitrile
Figure imgf000105_0003
[00430] To a suspension of 4-chloro-5-fluoro-pyridine-3-carbaldehyde oxime (6.8 g,
39.0 mmol) in dichloromethane (150 ml) was added carbonyl diimidazole (9.5 g, 58.5 mmol). The mixture was then heated at reflux for 30 minutes then cooled to room temperature, before being washed with saturated aqueous sodium bicarbonate followed by water. The organic layer was dried over sodium sulfate and concentrated in vacuo then the resultant residue triturated in diethyl ether / cyclohexane to afford the title compound as a pale yellow solid (4.05 g, 79%). 1H NMR (CDCl3400 MHz) 8.71 (IH, d, J= 0.4 Hz), 8.70 (IH, s). [00431] Step 3: 3-Amino-7-fluoro-furor3,2-c1pyridine-2-carboxylic acid ethyl ester
Figure imgf000106_0001
[00432] 4-Chloro-5-fluoronicotinonitrile (4.0 g, 25.6 mmol) was dissolved in DMF (50 ml) and treated with potassium carbonate (17.8 g, 128 mmol) followed by ethyl glycolate (3.64 ml, 38.4 mmol). The resultant reaction mixture was heated at 8O0C for 50 minutes, then cooled to room temperature and diluted with ethyl acetate. The solution was washed with water (x 2), dried (Na2SO4), filtered and concentrated in vacuo. The resultant solid was triturated in diethyl ether to give the title compound as an off-white solid (3.58 g, 63%). LCMS (method B): Rx = 2.65 min, M+H+ 225.
[00433] Step 4: 7-Fluoro-3-(2-fluoro-4-trimethylsilanyl-phenylamino)- furo|"3.2- cipyridine-2-carboxylic acid ethyl ester
Figure imgf000106_0002
[00434] A degassed solution of 3-amino-7-fluoro-furo[3,2-c]pyridine-2-carboxylic acid ethyl ester (2.5 g, 11.1 mmol), trifluoro-methanesulfonic acid 2-fluoro-4- trimethylsilanyl-phenyl ester (4.2 g, 13.3 mmol), Pd2dba3 (508 mg, 0.56 mmol), Xantphos (642 mg, 1.12 mmol) and Cs2CO3 (7.2 g, 22.2 mmol) in toluene (25 ml) was heated at reflux for 1 hour. The reaction mixture was cooled to ambient temperature then filtered through a pad of Celite ® washing with ethyl acetate. The filtrate was concentrated in vacuo and the resultant residue subjected to flash chromatography (Si-SPE, gradient 0-30% ethyl acetate in cyclohexane) to provide a red/orange residue. The residue was triturated in methanol to give the title compound as a yellow solid (2.5 g, 58%). LCMS (method B): Rx = 4.71 min, M+H+
391.
[00435] Step 5: 7-Fluoro-3-(2-fluoro-4-iodo-phenylamino)-furo|"3.2-c]pyridine- carboxylic acid ethyl ester
[00436] To a solution of 7-fluoro-3-(2-fluoro-4-trimethylsilanyl- phenylamino)furo[3,2c]-pyridine-carboxylic acid ethyl ester (2.5 g, 6.4 mmol) in dichloromethane (60 ml) at O0C was added iodine monochloride (2.08 g, 12.8 mmol, solution in dichloromethane) and the solution was stirred and allowed to warm for 45 minutes. The precipitated solid was filtered off, the residue retained, and the filtrate washed with saturated aqueous sodium thiosulfate, dried (Na2SO4), filtered and concentrated in vacuo to give a residue. The residues from filtration and concentration were combined and triturated in diethyl ether to give a pale tan solid (2.58 g, 91%). LCMS (method B): Rx = 4.14 min, M+H+
= 445.
[00437] 4-Chloro-3-(2-fluoro-4-iodo-phenylamino)-furo[3.2-c]pyridine-2-carboxyric acid ethyl ester
Figure imgf000107_0001
[00438] Step 1 : 2,4-Dichloro-nicotinic acid ethyl ester
Figure imgf000107_0002
[00439] To a solution of diisopropylamine (2.4 ml, 16.9 mmol) in THF (40 ml) at -
780C under inert atmosphere, was added n-butyllithium (10.6 ml, 16.9 mmol, 1.6M in hexanes) and the solution was stirred for 15 min at -780C. 2,4-Dichloropyridine (1.8 mL, 16.9 mmol) was added dropwise and the reaction mixture was stirred at -780C for 2 hours before the addition of ethyl cyanoformate (4.0 ml, 40.4 mmol). The reaction was then stirred at -780C for 1 hour and then allowed to warm to room temperature. The mixture was then partitioned between water and ethyl acetate and the layers separated. The organic layer was washed with a saturated solution of sodium bicarbonate flowed by brine, dried over magnesium sulfate, then concentrated in vacuo. Purification of the resultant residue by flash chromatography (Si-SPE, pentane : diethyl ether gradient 1:0 to 4: 1) afforded the title compound as colourless oil (1.6 g, 43%). 1H NMR (CDCl3, 400 MHz) 8.34 (IH, d, J= 5.4 Hz), 7.33 (IH, d, J= 5.4 Hz), 4.49 (2H, q, J= 7.1 Hz), 1.43 (3H, t, J= 7.1 Hz).
[00440] Step 2: 4-Chloro-3-hvdroxy-furor3,2-clpyridine-2-carboxylic acid ethyl ester
Figure imgf000108_0001
[00441] To a solution of 2,4-dichloro-nicotinic acid ethyl ester (1.6 g, 7.3 mmol) and ethyl glycolate (0.72 mL, 7.6 mmol) in DMF at 00C under an inert atmosphere was added sodium hydride (60% in mineral oil, 584 mg, 14.6 mmol) portionwise. The reaction mixture was stirred at 00C for 3 hours, quenched by careful addition of acetic acid (ca. 5 mL), diluted with water and extracted into ethyl acetate. The organic layer was separated, washed with water, then brine, dried over sodium sulphate and concentrated to give the title compound as a yellow solid (1.75 g, 100%). LCMS (method B): Rx = 2.99 min, M+H+= 242. [00442] Step 3: 4-Chloro-3-trifluoromethanesulfonyloxy-furo[3,2-clpyridine-2- carboxylic acid ethyl ester
Figure imgf000108_0002
[00443] A mixture of 4-chloro-3-hydroxy-furo[3,2-c]pyridine-2-carboxylic acid ethyl ester (1.8 g, 7.5 mmol), N-phenyl-trifluoromethanesulfonimide (5.0 g, 14.0 mmol) and diisopropylethyl amine (5.5 mL, 32.3 mmol) in dimethoxyethane (30 mL) was stirred at 900C for 48 hours, cooled to room temperature and concentrated under reduced pressure. The resultant residue was purified by flash chromatography (Si-SPE, cyclohexane : ethyl acetate, gradient 1: 0 to 1 : 1) to afford the title compound as a pale yellow solid (1.06 g, 38%). LCMS (method B): Rx = 3.94 min, M+H+ = 374.
[00444] Step 4: 4-Chloro-3-(2-fluoro-4-trimethylsilanyl-phenylamino)-furor3,2- cipyridine-2-carboxylic acid ethyl ester
Figure imgf000108_0003
[00445] A degassed solution of 4-chloro-3-trifluoromethanesulfonyloxy-furo[3,2- c]pyridine-2-carboxylic acid ethyl ester (810 mg, 2.17 mmol), 2-fluoro-4-trimethylsilanyl- phenylamine (306 mg, 1.67 mmol), Pd2dba3 (31 mg, 0.03 mmol), Xantphos (58 mg, 0.10 mmol) and cesium carbonate (817 mg, 2.50 mmol) in toluene (17 ml) was heated at reflux under an argon atmosphere for 16 hours. The reaction mixture was filtered through Celite ® and concentrated in vacuo. The resultant residue was purified by flash chromatography (Si- SPE, pentane: diethyl ether, gradient 1 : 0 to 0: 1) to afford the title compound as a white solid (558 mg, 82%). LCMS (method B): Rx = 4.64 min, M+H+ = 407.
[00446] Step 5: 4-Chloro-3-(2-fluoro-4-iodo-phenylamino)-furor3,2-c1pyridine-2- carboxylic acid ethyl ester
[00447] To a solution of 4-chloro-3-(2-fluoro-4-trimethylsilanyl-phenylamino)- furo[3,2-c]pyridine-2-carboxylic acid ethyl ester (265 mg, 0.65 mmol) in dichloromethane (6.5 ml) at O0C was added iodine monochloride (1.3 ml, 1.3 mmol, IM solution in dichloromethane) and the solution was stirred at this temperature for 1 hour. A saturated solution of sodium thiosulfate (5 ml) was added and the mixture was poured into saturated sodium thiosulfate (25 ml). The aqueous layer was extracted with dichloromethane (2 x 25 ml), the combined organic layers were washed with brine, dried over magnesium sulfate and concentrated in vacuo to yield the title compound as a yellow solid (239 mg, 80%). LCMS (method B): Rx = 4.22 min, M+H+ = 461.
[00448] 3-(2-Fluoro-4-iodo-phenylamino)-4-methyl-furor3.2-clpyridine-2-carboxylic acid ethyl ester
Figure imgf000109_0001
[00449] Step 1 : 3-(2-Fluoro-4-trimethylsilanyl-phenylamino)-4-methyl- furor3,21pyridine-2-carboxylic acid ethyl ester
Figure imgf000109_0002
[00450] To a solution of 4-chloro-3-(2-fluoro-4-trimethylsilanyl-phenylamino)- furo[3,2-c]pyridine-2-carboxylic acid ethyl ester (406 mg, 1.0 mmol) in dioxane (5 mL) were added trimethylboroxine (0.14 mL, 1.0 mmol), tetrakis(triphenylphosphine)palladium (115 mg, 0.1 mmol) and potassium carbonate (207 mg, 1.5 mmol) before the mixture was degassed and heated at reflux for 6 hours. The reaction mixture was cooled to room temperature and filtered through a pad of Celite ® which was washed with ethyl acetate. The filtrates were combined and concentrated in vacuo to give a residue which was purified by flash chromatography (Si-SPE, pentane : diethyl ether 1:0 to 0: 1). The title compound was obtained as a pale yellow oil (221 mg, 57%). LCMS (method B): Rx = 3.53 min, M+H+ =
387.
[00451] Step 2: 3-(2-Fluoro-4-iodo-phenylamino)-4-methyl-furor3,2-c1pyridine-2- carboxylic acid ethyl ester
[00452] To a solution of 3-(2-fluoro-4-trimethylsilanyl-phenylamino)-4-methyl- furo[3,2]pyridine-2-carboxylic acid ethyl ester (215 mg, 0.56 mmol) in dichloromethane (5 ml) at O0C was added iodine monochloride (1.1 ml, 1.1 mmol, IM solution in dichloromethane) and the solution was stirred at this temperature for 1 hour. A saturated solution of sodium thiosulfate (5 ml) was added and the mixture was poured into saturated sodium thiosulfate (25 ml). The aqueous layer was extracted with dichloromethane (2 x 25 ml), the combined organic layers were washed with brine, dried over magnesium sulfate and concentrated in vacuo. Purification of the resultant residue by flash chromatography (Si-SPE, cyclohexane : dichloromethane gradient 1 :0 to 0: 1) afforded the title compound as a yellow solid (241 mg, 98%). LCMS (method B): Rx = 2.99 min, M+H+ = 441.
[00453] 3-(2-Fluoro-4-methylsulfanyl-phenylamino)-furor3.2-clpyridine-2-carboxylic acid ethyl ester
Figure imgf000110_0001
[00454] 3-Trifluoromethanesulfonyloxy-furo[3,2-c]pyridine-2-carboxylic acid ethyl ester (1.26 g, 3.71 mmol) and 2-fluoro-4-methylsulfanyl-phenylamine (816 mg, 5.20 mmol) were dissolved in toluene (25 ml) and Pd2(dba)3 (170 mg, 0.19 mmol) was added, followed by Xantphos (214 mg, 0.37 mmol) and potassium phosphate tribasic (1.57 g, 7.42 mmol). The mixture was thoroughly degassed and purged with argon, then stirred under argon at 12O0C for 16 hours. After cooling, the mixture was filtered through Celite ® then concentrated. Purification of the resultant residue by flash chromatography (Si-SPE, etheπpentane gradient 1 :4 to 1 :0) gave the title compound as a tan solid (770 mg, 60%). LCMS (method B): Rx = 3.29, M+H+ 347.
[00455] 7-Fluoro-3-(2-fluoro-4-methylsulfanyl-phenylamino)-furo[3,2- c]pyridine-
2-carboxylic acid ethyl ester
Figure imgf000111_0001
[00456] A degassed solution of 3-amino-7-fluoro-furo[3,2-c]pyridine-2-carboxylic acid ethyl ester (0.2 g, 0.89 mmol), l-bromo-2-fluoro-4-methylsulfanyl-benzene (0.34 g, 1.5 mmol), Pd2dba3 (0.041 g, 0.045 mmol), Xantphos (0.052 g, 0.089 mmol) and K3PO4 (0.38 g, 1.8 mmol) in toluene (5 ml) was heated at reflux for 18 hours. The reaction mixture was cooled to ambient temperature then filtered through a pad of Hyflo washing with ethyl acetate. The filtrate was concentrated in vacuo and the resultant residue subjected to flash chromatography (Si-SPE, gradient 0-10% ethyl acetate in dichloromethane) to provide the title compound as a yellow solid (0.18 g, 55%). LCMS (method B): Rx = 3.95 min, M+H+ 365.
[00457] 7-Chloro-3-(2-fluoro-4-methylsulfanyl-phenylamino)-furor3,2-c1pyridine-2- carboxylic acid ethyl ester
Figure imgf000111_0002
[00458] A degassed solution of 3-amino-7-chloro-furo[3,2-c]pyridine-2-carboxylic acid ethyl ester (0.1 g, 0.42 mmol), l-bromo-2-fluoro-4-methylsulfanyl-benzene (0.16 g, 0.71 mmol), Pd2dba3 (0.019 g, 0.021 mmol), Xantphos (0.024 g, 0.042 mmol) and K3PO4 (0.18 g, 0.83 mmol) in toluene (2.5 ml) was heated at reflux for 18 hours. The reaction mixture was cooled to ambient temperature then filtered through a pad of Hyflo washing with ethyl acetate. The filtrate was concentrated in vacuo and the resultant residue subjected to flash chromatography (Si-SPE, gradient 0-10% ethyl acetate in dichloromethane) to provide the title compound as a pale yellow solid (0.087 g, 54%). LCMS (method B): Rx = 4.14 min, M+H+ 379.
[00459] SYNTHESIS OF REPRESENTATIVE AMINES AND HYDROXYLAMΓNES [00460] Cyclopropylmethylhydroxylamine Hydrochloride
^NH2 HCI
[00461] Prepared according to Marquez et al (2005) Synth. Comm. 35(17):2265-2269 [00462] O-((R)-2.2-dimethyl-[1.31dioxolan-4-ylmethyl)hydroxylamine
Figure imgf000112_0001
[00463] Prepared according to Bailey et al (1991) J. Med. Chem. 34(l):57-65 [00464] O-(2-Vinyloxy-ethyl)-hvdroxylamine
,NH2
O
[00465] Prepared according to WO 0206213 [00466] N-Methyl-O-(2-vinyloxy-ethyl)-hvdroxylamine
Figure imgf000112_0002
[00467] Formaldehyde (37% w/w in water, 80 μL, 1.0 mmol) was added to a cooled
(O0C) solution of O-(2-vinyloxy-ethyl)-hydroxylamine (105 mg, 1.0 mmol) in ethanol (1 mL). The mixture was stirred for 30 minutes before addition of pyridiniumpαrα-toluene sulfonate (250 mg, 1.0 mmol) and sodium cyanoborohydride (70 mg, 1.1 mmol). The resultant suspension was allowed to warm to ambient temperature and stirred for 20 hours. The solvent was evaporated and the residue diluted with ethyl acetate (25 mL) and washed with brine (20 mL), dried (MgSO4), filtered, then evaporated to provide the desired product as an oil (84 mg, 71%). IH NMR (CDCl3, 400MHz) 6.44-6.55 (m, IH), 4.98 (s, IH), 4.16-4.24 (m, IH), 3.98-4.06 (m, IH), 3.82-3.96 (m, 4H), 2.59 (s, 3H). [00468] 4-(tert-Butyl-dimethyl-silanyloxy)-isoxazolidine
Figure imgf000112_0003
[00469] tert-Butyl-dimethyl-chlorosilane (0.5 g, 3.21 mmol) was added to a stirred solution of isoxazolidin-4-ol hydrochloride (0.40 g, 3.18 mmol) in DMF (3 mL) and the mixture left to stir at ambient temperature for 2.5 hours. The solvent was evaporated and the residue partitioned between ethyl acetate (50 mL) and water (20 mL). The organic phase was separated, washed with water (3 x 20 mL) then brine (20 mL), dried (MgSO4), filtered and evaporated to provide the desired product as a colourless oil (0.62 g, 96%). IH NMR (CDCI3, 400MHz) 5.52 (s, IH), 4.60-4.65 (m, IH), 3.45-3.62 (m, IH), 3.80-4.05 (m, IH), 2.80-3.05 (m, 2H), 0.80 (s, 9H), 0.08 (s, 6H). [00470] (S)-3-Aminooxy-pyrroridine- 1-carboxylic acid tert-butylester
Figure imgf000113_0001
[00471] Step 1: (S)-3-d.3-Dioxo-1.3-dihvdro-isoindol-2-yloxy)-pyrrolidine-l- carboxylic acid tert-butyl ester
Figure imgf000113_0002
[00472] (R)-3 -Hydroxy-pyrrolidine- 1-carboxylic acid tert-butyl ester (1.37 g, 7.31 mmol) was dissolved in THF (20 mL), 2-hydroxy-isoindole-l,3-dione (1.19 g, 7.31 mmol) and triphenyl phosphine (1.92 g, 7.31 mmol) were added followed by dropwise addition of diisopropyl azodicarboxylate (1.33 mL, 8.04 mmol) over 10 minutes. The reaction mixture was allowed to stir at ambient temperature for 18 hours then the solvent was evaporated. The residue was purified by flash column chromatography (Si-SPE, DCM: EtOAc, gradient 100:0 to 80:20) to provide the title compound as a colourless oil (1.43 g, 59%). IH NMR (CDCl3, 400MHz) 7.86 (m, 2H), 7.77 (m , 2H), 4.94-5.02 (m, IH), 3.66-3.84 (m, 2H), 3.50-3.65 (m, 2H), 2.24-2.32 (m, IH), 1.93-2.05 (m, IH), 1.49 (s, 9H).
[00473] Step 2: (S)-3-Aminooxy-pyrrolidine- 1-carboxylic acid tert-butylester
[00474] Methyl hydrazine (0.23 mL, 4.40 mmol) was added dropwise over 5 minutes to a solution of (S)-3-(l,3-dioxo-l,3-dihydro-isoindol-2-yloxy)-pyrrolidine-l-carboxylic acid tert-butyl ester (1.43 g, 4.3 mmol) in DCM (12 mL). The mixture was stirred at ambient temperature for 1 hour then evaporated. The residue was suspended in diethyl ether (10 mL) and the solid was filtered. The filtrate was concentrated to provide the title compound as a colourless oil (0.86 g, 99%).1H NMR (CDCl3, 400MHz) 4.24-4.26 (m, IH), 3.60-3.66 (m, IH), 3.44-3.54 (m, IH), 3.30-3.42 (m, 2H), 2.03-2.12 (m, IH), 1.84-1.96 (m, IH), 1.46 (s, 9H). [00475] 2-Aminooxy-2-methyl-propan- 1 -ol hydrochloride HC|1 Λr^H
[00476] Step 1 : 2-(N-Boc-aminooxy)isobutyric acid ethyl ester
Figure imgf000114_0001
[00477] To a solution of N-Boc-hydroxylamine (5.2 g, 39.05 mmol) in ethanol (100 mL) was added potassium hydroxide (2.6294 g, 46.86 mmol) and stirred at room temperature till the potassium hydroxide dissolved into solution. To this was added 2-Bromoisobutyric acid ethyl ester (6.87 mL, 46.86 mmol) and refluxed overnight. A white precipitate was observed after 1 hour. Reaction cooled to room temperature and then filtered. The white solid was discarded and the filterate was concentrated. The oily residue was partitioned between water (75 mL) and ether (3 X 100 mL). The combined ether layer was dried with sodium sulfate, filtered and the filterate was concentrated to give the title compound as a clear oil (9.543 g, 99%). LCMS (method C): Rx = 2.55 min, M+H+ = 247.9. IH NMR (CDCl3, 400MHz) 4.20 (q, 2H), 1.50 (s , 6H), 1.498 (s, 9H), 1.30 (t, 3H). [00478] Step 2: 2-(N-Boc-aminooxy)-2-methylpropan-l-ol
Figure imgf000114_0002
[00479] To a solution of 2-(N-Boc-aminooxy)isobutyric acid ethyl ester (2.35 g, 9.5 mmol) in anhydrous ethyl ether (100 mL) at 00C under nitrogen was added 1.0 M Lithiumtetrahydroaluminate in tetrahydrofuran (17.106 mL, 17 mmol) and stirred at 00C under nitrogen for 5 hours. To this was added a couple of CO2 pellets (dry ice), followed by water (25 mL) at 00C. This was then stirred overnight and let warm to room temperature in the process. The ether layer was decanted off and kept aside. The white solid was triturated with ether and the ether was combined with the ether layer obtained before. The white solid was then discarded. The combined ether layers was dried with sodium sulfate, filtered and concentrated to give the title compound as a white solid ( 1.94 g, 99.5%). IH NMR (CDCI3, 400MHz) 3.40 (s, 2H), 1.50 (s , 9H), 1.20 (s, 6H). [00480] Step 3: 2-Aminooxy-2-methyl-propan-l-ol hydrochloride
[00481] To a solution of 2-(N-Boc-aminooxy)-2-methylpropan-l-ol (1.94 g, 9.45 mmol) in anhydrous dichloromethane (10 mL) was added 4 M HCl in dioxane (47.26 mL, 200 mmol) at room temperature and stirred for 1 hour. The reaction was concentrated under reduced pressure and the residue triturated with ether (3 X 30 mL) to give the title compound as an oil/white solid (HCl salt). The oil/white solid was dried under vacuum and used as is for the coupling step. (1.10 g, 82.2 %). IH NMR (DMSO-d6, 400MHz) 3.58 (s, 2H), 3.48 (s, 2H), 1.34 (s , 6H). [00482] 1 -Aminooxy-2-methylpropan-2-ol
H
H_ >C
O
[00483] Step 1 : 2-(2-Hvdroxy-2-methyl-propoxy)-isoindole- 1 ,3 -dione
Figure imgf000115_0001
[00484] To a solution ofN-Hydroxyphthalimide (18.3 g, 112 mmol) and l,2-Epoxy-3- methyl propane (9.50 mL, 107 mmol) in anhydrous DMF under nitrogen at room temperature was added triethylamine (16.1 mL, 115 mmol). The reaction turned from yellow to dark red. The reaction was then heated to 85°C overnight. The reaction was cooled to room temperature and concentrated under reduced pressure. The residue obtained was partitioned between water (100 mL) and ether (3 X 75 mL). The combined ether layers were washed with water (2 X 50 mL), dried with anhydrous magnesium sulfate, filtered and concentrated to give a yellow oil (26.8 g). This was then treated with dichloromethane (35 mL), which resulted in unreacted N-Hydroxyphthalimide crashing out as a white precipitate. This was filtered off and discarded. The fϊlterate was purified by flash column chromatography (120 g, silica, ISCO, 45 mL/min, 0-10% methanol in dichloromethane in 50 minutes) to give the title compound as a white solid (13.4 g, 53.3 %). LCMS (method C): Rτ = 1.70 min, M+H+ = 236.1 IH NMR (CDCl3, 400MHz) 7.84 (m, 2H), 7.78 (m, 2H), 4.15 (s, 2H), 1.39 (s, 6H). [00485] Step 2: l-Arninooxy-2-methylpropan-2-ol
[00486] To a solution of 2-(2-Hydroxy-2-methyl-propoxy)-isoindole-l,3-dione (3,70 g,
15.7 mmol) in anhydrous dichloromethane (25 mL) under nitrogen at 00C was added methyl hydrazine (0.879 mL, 16.50 mmol) and stirred for 2 hours at 00C. The addition of methyl hydrazine resulted in a pale yellow color followed by a white precipitate. The reaction was filtered after 2 hours at 00C and the solid was discarded. The filtrate was concentrated under reduced pressure to give the title compound as a pale yellow oil (1.65 g, 100%). LCMS (method C): Rx = 0.34 min, M+H+ = 106.1. IH NMR (DMSO-d6, 400MHz) 3.60 (s, 2H), 1.22 (s , 6H). [00487] 3 -aminooxy-3 -methylbutan- 1 -ol
Figure imgf000116_0001
[00488] Step 1: 2-(3-Hvdroxy-3-methyl-butoxy)-isoindole-1.3-dione
Figure imgf000116_0002
To a solution of N-Hydroxyphthalimide (3.13 g, 19.2 mmol) and 3-Hydroxy-3-methyl butane (2.0O g, 19.2 mmol) in anhydrous dichloromethane under nitrogen at room temperature was added Boron trifluoride etherate (2.43 mL, 19.2 mmol) and stirred overnight. The reaction turned black after 18h with a white precipitate (N-Hydroxyphthalimide). To the reaction was added saturated sodium bicarbonate solution (3 mL) and stirred for 5 minutes at room temperature. The reaction was then filtered and the white solid was discarded. The filtrate was concentrated and the black residue obtained was re-dissolved in 25 mL dichlormethane and filtered. The white solid was discarded and the filtrate was concentrated to give a black residue. This was dissolved in dichloromethane (5 mL) and purified by flash column chromatography ( silica, 80 g, ISCO, 30 ml/min, 0-100% ethyl acetate in hexane in 45 minutes) to give the title compound as a yellow oil (228 mg, 4.75%). LCMS (method C): RT = 1.77 min, M+H+ = 250.2. IH NMR (CDCl3, 400MHz) 7.83 (m, 2H), 7.78 (m, 2H), 3.95 (t, 2H), 2.00 (t, 2H), 1.45 (s, 6H).
[00489] Step 2: 3-aminooxy-3-methylbutan-l-ol
[00490] To a solution of 2-(3-Hydroxy-3-methyl-butoxy)-isoindole-l,3-dione (228 mg,
0.91 mmol) in anhydrous dichloromethane (2 mL) under nitrogen at 00C was added methyl hydrazine (0.05 mL, 0.96 mmol) and stirred for 1 hour and let warm to room temperature in the process. The addition of methyl hydrazine resulted in a pale yellow color followed by a white precipitate. The reaction was filtered after 2 hours at 00C and the solid was discarded. The filtrate was concentrated under reduced pressure to give the title compound as a pale yellow solid (95 mg, 87%). LCMS (method C): Rx = 0.34 min, M+H+ = 120. IH NMR (DMSO-d6, 400MHz) 3.75 (t, 2H), 1.83 (t, 2H), 1.24 (s , 6H). [00491] O-Pyridin-2-ylmethyl-hvdroxylamine hydrochloride
Figure imgf000116_0003
[00492] Step 1 : N-Boc-aminooxymethyl(pyridine-2-yl)
Figure imgf000117_0001
[00493] To a solution of N-Boc-hydroxylamine (5.0 g, 37.6 mmol) in ethanol (100 mL) was added potassium hydroxide (4.63 g, 82.61 mmol) and stirred at room temperature till the potassium hydroxide dissolved into solution. To this was added 2- Bromomethylpyridine hydrobromide (11.398 g, 45.06 mmol) and refluxed overnight. A white precipitate was observed after 1 hour. Reaction cooled to room temperature and then filtered. The white solid was discarded and the fϊlterate was concentrated. The oily residue was partitioned between water (75 mL) and ether (3 X 100 mL). The combined ether layer was dried with sodium sulfate, filtered and the filterate was concentrated to give the product as a yellow oil (6.0 g). The oil was dissolved in dichloromethane (10 mL) and purified by flash column chromatography (silica, 120 g, ISCO, 45 mL/min, 0-100 % ethyl acetate in hexane in 40 minutes) to give the title compound as a white solid (1.78g, 21.2%). LCMS (method C): RT = 1.13 min, M+H+ = 225.2. IH NMR (CDCl3, 400MHz) 8.61 (d,m, IH), 7.73 (t,d, 1 H), 7.52 (s, IH), 7.46 (d,t, IH), 5.01 (s, 2H), 1.50 (s , 9H). [00494] Step 2: O-Pyridin-2-ylmethyl-hvdroxylamine hydrochloride
[00495] To a solution of N-Boc-aminooxymethyl(pyridine-2-yl) (860 mg, 3.8 mmol) in anhydrous dichloromethane (2 mL) was added 4 M HCl in dioxane (5.06 mL, 20 mmol) at room temperature and stirred for 2 hours. To the reaction was added ether (25 mL) and stirred for 5 minutes. The solvents were decanted off and the residue was treated with ether (25 mL), followed by stirring and then decanting again. This was repeated one more time and the residue (white solid) was dried under vacuum to give the title compound as a white solid ( 688 mg, 91%). LCMS (method C): Rx = 0.36 min, M+H+ = 125.0. IH NMR (DMSO-d6, 400MHz) 8.70 (m, IH), 8.05 (m, IH), 7.60 (m, 2H), 5.20 (s, 2H). [00496] Q-(I -Phenyl-ethyD-hydroxylamine hydrochloride
Figure imgf000117_0002
[00497] Synthesized from 1 -(bromoethyl)benzene following procedures analogous to those used to synthesize O-Pyridin-2-ylmethyl-hydroxylamine hydrochloride.
[00498] LCMS (method C): Rx = 0.92 min, M+H+ = 138.2. IH NMR (DMSO-d6, 400MHz) 10.90 (s, 2H), 7.45 (m, 5H), 5.25 (q, IH), 1.50 (d, 3H).
[00499] O-r2-(ter?-Butyl-dimethyl-silanyloxy)-propyl1-hvdroxylamine
-Sl~O^%H2
[00500] Step 1 : (2-Benzyloxy-l-methyl-ethoxy)-ter?-butyl-dimethyl-silane
>u
[00501] Tert-butyldimethylsilyl chloride (517 mg, 3.43 mmol) was added to a solution of l-benzyloxy-propan-2-ol (518 mg, 3.12 mmol), imidazole (318 mg, 4.66 mmol), and A- DMAP (95 mg, 0.78 mmol) in CH2Cl2 (3 mL). The reaction mixture was stirred at room temperature for 16 h, then 2g of silica gel was added and the volatiles were removed in vacuo. The residue was purified by silica chromatography (0 - 5% EtOAc :Hex) to afford the title compound (713 mg, 82% yield) as a clear oil. [00502] Step 2: 2-(fert-Butyl-dimethyl-silanyloxy)-propan-l-ol
;suo> — OH
[00503] To a solution of (2-Benzyloxy- 1 -methyl-ethoxy)-ter?-butyl-dimethyl-silane
(640 mg, 2.3 mmol) in ethyl acetate (10 mL) was added 20% Pd on carbon (64 mg). The reaction mixture was evacuated and flushed with H2, then stirred under an atmosphere of H2 for 3h. The reaction mixture was then filtered through celite and concentrated to afford the title compound (430 mg, 99% yield) as a clear oil, which was used without further purification in the next step. [00504] Step 3 : 2-|"2-(ferf-Butyl-dimethyl-silanyloxy)-propoxy"|-isoindole- 1 ,3-dione
Figure imgf000118_0001
[00505] DEAD (0.46 mL, 2.94 mmol) was added dropwise to a solution oϊ2-(tert-
Butyl-dimethyl-silanyloxy)-propan-l-ol (430 mg, 2.26 mmol), triphenylphosphine (593 mg, 2.26 mmol), and N-hydroxyphthalimide (369 mg, 2.26 mmol) in THF (10 mL) at 0 0C. After stirring for 10 minutes at 0 0C, the reaction mixture was brought to room temperature and stirring was continued for a further 48 h. The reaction mixture was filtered through a coarse glass funnel and concentrated in vacuo. The residue was purified by silica chromatography (0 - 40% EtOAc: Hex) to afford the title compound (139 mg, 18% yield) as a white solid. [00506] Step 4: O-r2-(ter?-Butyl-dimethyl-silanyloxy)-propyl1-hvdroxylamine
[00507] N-methylhydrazine (23 μL, 0.43 mmol) was added to a solution of 2-[2-(tert- Butyl-dimethyl-silanyloxy)-propoxy]-isoindole-l,3-dione (135 mg, 0.40 mmol) in CH2CI2 (3 mL). After stirring for Ih at room temperature, the white precipitate was filtered off and the reaction mixture was concentrated in vacuo to afford the title compound (76 mg, 92% yield) as a yellow oil. 1H NMR (CDCl3, 400MHz) 5.48 (br, 2H), 4.04 (m, IH), 3.58 (dd, IH), 3.52 (dd, IH), 1.13 (d, 3H), 0.89 (s, 9H), 0.09 (s, 6H). [00508] O-r2-(ter?-Butyl-dimethyl-silanyloxy)-l-methyl-ethyl1-hvdroxylamine
Figure imgf000119_0001
[00509] Step 1 : l-(ter?-Butyl-dimethyl-silanyloxy)-propan-2-ol
Figure imgf000119_0002
[00510] Tert-butyldimethylsilyl chloride (4.1 g, 27 mmol) was added to a solution of propane- 1,2-diol (2.0 mL, 27 mmol) and triethylamine (4.93 mL, 35.4 mmol) in CH2CI2. After stirring overnight at room temperature, the reaction mixture was washed one time eachwith IN aqueous HCl solution, water, and a 1: 1 saturated solution OfNaHCO3 and brine. The organic layer was dried over Na2SO4, then filtered and concentrated. The crude title compound was used without further purification in the next step.
[00511] Step 2: 2-r2-(fer^Butyl-dimethyl-silanyloxy)-l-methyl-ethoxy1-isoindole-l,3- dione
Figure imgf000119_0003
[00512] DEAD (1.86 mL, 11.8 mmol) was added dropwise to a solution of \-(tert-
Butyl-dimethyl-silanyloxy)-propan-2-ol (1.73 g, 9.09 mmol), triphenylphosphine (2.38 g, 9.09 mmol), and N-hydroxyphthalimide (1.48 g, 9.09 mmol) in THF (45 mL) at 0 0C. After stirring for 10 minutes at 0 0C, the reaction mixture was brought to room temperature and stirring was continued for a further 48 h. The reaction mixture was filtered through a coarse glass funnel and concentrated in vacuo. The residue was purified by silica chromatography (0 - 40% EtOAc:Hex) to afford the title compound (1.80 g, 59% yield) as a clear oil. [00513] Step 3 : O- [2-(ter?-Butyl-dimethyl-silanyloxy)- 1 -methyl-ethyl] -hydroxylamine
[00514] N-methylhydrazine (310 μL, 5.74 mmol) was added to a solution of 2-[2-(tert-
Butyl-dimethyl-silanyloxy)-l-methyl-ethoxy]-isoindole-l,3-dione (1.80 g, 5.36 mmol) in CH2CI2 (20 mL). After stirring for Ih at room temperature, the white precipitate was filtered off and the reaction mixture was concentrated in vacuo to afford the title compound (682 mg, 62% yield) as a yellow oil. 1H NMR (CDCl3, 400MHz) 5.39 (br, 2H), 3.77 - 3.68 (m, IH), 3.67 (dd, IH), 3.61 (dd, IH), 1.13 (d, 3H), 0.90 (s, 9H), 0.08 (s, 6H). [00515] O-(2-Phenyl-1.3-dioxinan-5-yl)-hydroxylamine
Figure imgf000120_0001
[00516] Step 1 : 2-(2-Phenyl-1.3-dioxinan-5-yloxy)-isoindole-1.3-dione
Figure imgf000120_0002
[00517] Diethyl Azodicarboxylate (0.85 mL, 5.41 mmol) added dropwise to a solution of 2-Phenyl-l,3-dioxinan-5-ol (750 mg, 4.16 mmol), Triphenylphosphine (1.09 g, 4.16 mmol; ), and N-Hydroxyphthalimide (0.679 g, 4.16 mmol; ) in THF (20 mL) at 0 0C. Stirred overnight at 0 0C - room temperature, then concentrated in vacuo. Diluted with CH2Cl2, then filtered through a Whatman syringe filter. 4 g silica gel added and concentrated in vacuo. The residue was purified by silica chromatography (30 - 70% EtOAc : Hex, flush with 100% EtOAc) to afford the title compound (495 mg, 37% yield) as a white solid. [00518] Step 2: O-(2-Phenyl-l,3-dioxinan-5-yl)-hvdroxylamine
[00519] N-methylhydrazine (87 μL, 5.74 mmol) was added to a solution of 2-(2-
Phenyl-l,3-dioxinan-5-yloxy)-isoindole-l,3-dione (495 mg, 1.52 mmol) in CH2Cl2 (10 mL). After stirring for 3h at room temperature, the white precipitate was filtered off and the reaction mixture was concentrated in vacuo to afford the title compound (272 mg, 92% yield) as a yellow oil. 1H ΝMR (CDCl3, 400MHz) 7.50 - 7.46 (m, 2H), 7.40 - 7.35 (m, 3H), 5.44 (br, 2H), 5.41 (s, IH) 4.48 - 4.42 (m, 2H), 4.01 - 3.93 (m, IH), 3.66 - 3.60 (m, 2H). [00520] Ν-(2-Aminooxy-ethyl)-methanesulfonamide
H
H2N-O' 3-v
[00521] Step 1 : r2-(l,3-Dioxo-l,3-dihvdro-isoindol-2-yloxy)-ethvH-carbamic acid tert- butyl ester
Figure imgf000120_0003
[00522] To a suspension of N-(tert-butoxycarbonyl)ethanolamine (5.0 g, 31.0 mmol),
N-hydroxyphthalimide (5.1g, 31.0 mmol) and triphenylphosphine (8.5 g, 32.6 mmol) in tetrahydrofuran (30 ml) at 0 0C was added dropwise diisopropyl azodicarboxylate (6.3 ml, 32.6 mmol). The reaction was stirred and allowed to warm to room temperature over 16 hours. The reaction was concentrated in vacuo and the product purified by flash chromatography (SiC^, gradient ethyl acetate:cyclohexane, 20:80 to 30:70) to yield the title compound as an oil (14.2 g). 1H NMR (CDCl3, 400 MHz) 7.87-7.85 (2H, m), 7.79-7.77 (2H, m), 4.26 (2H, t, J= 5.5 Hz), 3.47-3.43 (2H, m), 1.47 (9H, s).
[00523] Step 2: 2-(2-Amino-ethoxy)-isoindole-l,3-dione
Figure imgf000121_0001
[00524] [2-(l,3-Dioxo-l, 3-dihydro-isoindol-2-yloxy)-ethyl]-carbamic acid tert-butyl ester (4.4 g, -8.6 mmol) was dissolved in hydrochloric acid in dioxane (4N, 20 ml) and stirred at room temperature for 3 hours. The reaction was concentrated in vacuo. The resultant residue was dissolved in ethyl acetate (20 ml) and the solution washed with sodium hydroxide solution (20 ml, IN). The aqueous layer was isolated then extracted with ethyl acetate (2 x 10 ml). The combined organic layers were washed with brine and then dried over magnesium sulfate and concentrated in vacuo to yield the title compound as a colourless oil (1.96 g). 1H NMR (CDCl3, 400 MHz) 7.85 (2H, dd, J= 5.4, 2.9 Hz), 7.72 (2H, dd, J= 5.4, 3.0 Hz), 3.99-3.97 (2H, m), 3.86-3.83 (2H, m).
[00525] Step 3: N-r2-α.3-Dioxo-1.3-dihvdro-isoindol-2-yloxy)-ethyl1- methanesulfonamide
Figure imgf000121_0002
[00526] To a solution of 2-(2-amino-ethoxy)-isoindole-l,3-dione (1.96 g, 8.1 mmol) in acetonitrile (20 ml) at 0 0C was added simultaneously methane sulfonylchloride (0.63 ml, 8.1 mmol) and triethylamine (2.3 ml, 16.2 mmol). The reaction was stirred at 0 0C for 1 hour then at room temperature for 1 hour. The reaction was filtered and the filtrate concentrated in vacuo. The resultant residue was dissolved in ethyl acetate (20 ml) and washed with water (20 ml). The aqueous layer was isolated then extracted with ethyl acetate (2 x 10 ml). The combined organic layers were washed with brine, dried over magnesium sulfate and concentrated in vacuo to yield the title compound as a white solid (1.2 g, 44%). 1H NMR (CDCl3, 400 MHz) 7.87 (2H, dd, J= 5.5, 3.1 Hz), 7.79 (2H, dd, J= 5.5, 3.2 Hz), 4.36 (2H, dd, J= 4.82, 4.62 Hz), 3.43-3.47 (2H, m), 3.05 (3H, s).
[00527] Step 4: N-(2-Aminooxy-ethyl)-methanesulfonamide [00528] To a suspension of N-[2-(l,3-dioxo-l,3-dihydro-isoindol-2-yloxy)-ethyl]- ethanesulfonamide (0.55 g, 1.92 mmol) in dichloromethane (15 ml) was added methylhydrazine (0.1 ml, 1.92 mmol). The reaction was stirred at room temperature for 30 minutes, over which time a white precipitate formed. The reaction was filtered and the filtrate concentrated in vacuo to give a residue. The residue was purified by flash chromatography (Siθ2, gradient 1-5% methanol in dichloromethane) to yield the title compound as a white solid (204 mg, 68%). 1H NMR (CDCl3, 400 MHz) 3.80 (2H, t, J = 4.9 Hz), 3.39 (2H, t, J= 4.8 Hz), 3.00 (3H, s). [00529] N-Cyclopropylmethyl-O-(2-vinyloxy-ethyl)-hydroxylamine
Figure imgf000122_0001
[00530] A solution of O-(2-vinyloxy-ethyl)-hydroxylamine (210 mg, 2.0 mmol) and cyclopropane carboxaldehyde (140 mg, 2.0 mmol) in ethanol (2.0 ml) was cooled to O0C under a nitrogen atmosphere then pyridinium p-toluenesulfonic acid (0.5 g, 2.0 mmol) and sodium cyanoborohydride (0.15 g, 2.2 mmol). The resultant mixture was stirred at ambient temperature for 24 hours. The reaction mixture was diluted with ethyl acetate, washed with water then brine, dried (TS^SO4), filtered and concentrated in vacuo to give the a colourless oil which was used crude in the subsequent step. [00531] O- r 1 -(Toluene-4-sulfonyl)- 1 H-imidazol-2-ylmethyli -hydroxylamine
H,
[00532] Step 1 : 2-ri-(Toluene-4-sulfonyl)-lH-imidazol-2-ylmethoxy1-isoindole-l,3- dione
Figure imgf000122_0003
[00533] Diisopropyl azodicarboxylate was added dropwise to a cooled (00C) solution of 2-(hydroxymethyl)-l-(p-tolylsulfonyl)imidazole (0.60 g, 2.4 mmol), triphenylphosphine (0.65 g, 2.5 mmol) and N-hydroxyphthalimide (0.39 g, 2.4 mmol) in THF (20 ml). The reaction was stirred and allowed to warm to room temperature over 40 hours. The reaction was concentrated in vacuo and the residue dissolved in dichloromethane (20 ml) causing the product to precipitate as a white solid. The product was collected by filtration and washed with dichloromethane (5 ml) to yield the title compound as a white solid (580 mg, 61%). LCMS (method B): Rx = 3.46 min, M+H+ = 398.
[00534] Step 2 : Q-[I -(Toluene-4-sulfonyD- 1 H-imidazol-2-ylmethyl] -hydroxylamine
[00535] Methylhydrazine (40 μl, 0.75 mmol) was added to a solution of 2-[l-(toluene-
4-sulfonyl)-lH-imidazol-2-ylmethoxy]-isoindole-l,3-dione (300 mg, 0.75 mmol) in dichloromethane (3 ml) and the reaction stirred at room temperature for 20 minutes. After approximately 10 minutes a white precipitate formed. The reaction was filtered and the filtrate concentrated in vacuo to approximately half the volume. Diethyl ether (5 ml) was added causing a white precipitate to form. The reaction was filtered and the filtrated concentrated in vacuo to give the title compound as a colourless oil (230 mg, 114%). The product was used without further purification. LCMS (method B): RT = 2.46 min, M+H+ = 268. [00536] (3 S.4S)-pyrrolidine-3.4-diol hydrochloride
HO, OH H2CI
[00537] Step 1 : GR.4R)-l-benzyl-3.4-dihvdroxypyrrolidine-2.5-dione
Figure imgf000123_0001
[00538] L-(+)-tartaric acid (1.51 g, 10.06 mmol) and benzyl amine (1.08 g, 10.06 mmol) in m-xylene (50 ml) were heated at reflux while water was collected in a Dean-Stark trap. After stirring overnight, the reaction was concentrated. The residue was taken up in minimal THF/EtOH and purified by silica gel flash chromatography (gradient elution, using 3: 1 hexane-ethyl acetate, ethyl acetate, and 9: 1 ethyl acetate-ethanol) to provide the title compound as a brown solid (0.99 g, 44%). [00539] Step 2: (3S.4S)-l-benzylpyrrolidine-3.4-diol
Figure imgf000123_0002
[00540] The (3R,4R)-l-benzyl-3,4-dihydroxypyrrolidine-2,5-dione (0.98 g, 4.4 mmol) in THF (20 ml) was added slowly to a stirring solution Of LiAlH4 (4.75 ml, 11.87 mmol of 2.5 M solution in THF) in THF cooled to -5°C. After complete addition, the reaction was warmed to room temperature, then heated at reflux overnight. The reaction was cooled to room temperature, then quenched with saturated aqueous NH4Cl until further addition produced no more bubbling. The reaction was diluted with ethyl acetate (20 ml), filtered, and the solid washed with ethyl acetate. The combined filtrates were concentrated, and the residue purified by silica flash chromatography (gradient elution, using EtOAc, and 9: 1 EtOAc-EtOH) to provide the title compound as a tan solid (0.52 g, 61%). [00541] Step 3: (3 S.4S)-pyrrolidine-3.4-diol hydrochloride
[00542] The (3S,4S)-l-benzylpyrrolidine-3,4-diol (0.52 g, 2.7 mmol) was dissolved in ethanol (15 ml) and acetic acid (10 ml) and hydrogenated (50 psi H2) over 10% Pd-C (100 mg) on a Parr apparatus for 6 hours. After filtering through Celite, and washing the filter cake with ethyl acetate, the combined filtrate and washings were concentrated. The residue was diluted with 4N HCl/dioxane (2 ml), methanol (5 ml), then toluene (40 ml) and concentrated. The residue was triturated with ethyl ether to provide the hydrochloride salt of the title compound as a tan solid (0.37 g, 97%). 1H NMR (D2O, 400MHz) 4.35 (d, J=3.4 Hz, 2H), 3.54 (dd, J = 12.8 Hz, 3.4 Hz, 2H), 3.30 (d, J = 12.8 Hz, 2H). [00543] 3 -methylpyrrolidin-3-ol hydrochloride
Figure imgf000124_0001
[00544] Step 1 : tert-butyl 3-hydroxy-3-methylpyrrolidine-l-carboxylate
Figure imgf000124_0002
[00545] The solution of (0.070 g, 0.38 mmol) tert-butyl 3-oxopyrrolidine-l- carboxylate in anhydrous THF (2 mL) was cooled to -78°C. Then the solution of 1 M methylmagnesium bromide in butyl ether was added dropwise. The reaction was stirred at - 78°C for 4 h and quenched by water (2 mL). After concentrating the reaction in vacuo, the residue was partitioned between ethyl acetate and water. The aqueous layer was extracted once more with ethyl acetate, and the combined organic layers were washed with brine, dried (MgSO4) and concentrated. The residue was purified by silica flash chromatography (gradient elution, using 1 : 1 hexane-ethyl acetate and ethyl acetate) to provide the title compound (0.054 g, 70%). [00546] Step 2: 3-methylpyrrolidin-3-ol hydrochloride
[00547] To tert-butyl S-hydroxy-S-methylpyrrolidine-l-carboxylate (0.027 g, 0.13 mmol) was added a 4N HCl/dioxane solution (1 ml) and the mixture was stirred for 2 hours.
The solution was concentrated in vacuo. The residue was diluted with toluene (1 ml) and reconcentrated to provide the title compound as a colorless oil (0.018 g, 100%).
[00548] (9H-fluoren-9-yl)methyl (3R.4R)-4-hvdroxypyrrolidin-3-ylcarbamate hydrochloride
HO
Fmo fc ~O HllCl
[00549] Step 1 : (9H-fluoren-9-vnmethyl GR.4R)-l-αert-butoxycarbonvn-4- hvdroxypyrrolidin-3 -ylcarbamate
Figure imgf000125_0001
[00550] (3R,4R)-tert-butyl 3-amino-4-hydroxypyrrolidine-l-carboxylate (0.05 g, 0.25 mmol) was dissolved in 1,4-dioxane (1 mL), water (1 mL), and toluene (0.3 mL). Then 9- fluorenylmethyl chloroformate (0.077 g, 0.30 mmol) was added slowly followed by sodium bicarbonate (0.083 g, 0.99 mmol). The reaction mixture was stirred at room temperature overnight. After concentrating the reaction in vacuo, the residue was partitioned between ethyl acetate and water. The aqueous layer was extracted once more with ethyl acetate, and the combined organic layers were washed with brine, dried (MgSO4) and concentrated. The residue was purified by silica flash chromatography (gradient elution, using 1 : 1 hexane-ethyl acetate and ethyl acetate) to provide the title compound (0.090 g, 90%).
[00551] Step 2: (9H-fluoren-9-vnmethyl GR.4R)-4-hvdroxypyrrolidin-3-ylcarbamate hydrochloride
[00552] A 4N HCl/dioxane solution (1 ml) was added to (9H-fluoren-9-yl)methyl
(3R,4R)-l-(tert-butoxycarbonyl)-4-hydroxypyrrolidin-3-ylcarbamate and the mixture was stirred for 2 hours. The solution was concentrated in vacuo. The residue was diluted with toluene (1 ml) and reconcentrated to provide the title compound as a colorless oil (0.076 g,
100%).
[00553] (2R.3Ry2-(hvdroxymethvnpyrrolidin-3-ol hydrochloride
Figure imgf000126_0001
[00554] Step 1 : (2S.3R)-l-(tert-butoxycarbonyl)-3-hydroxypyrrolidine-2-carboxylic acid
Figure imgf000126_0002
[00555] The (2S,3R)-l-(tert-butoxycarbonyl)-3-hydroxypyrrolidine-2-carboxylic acid
(1.76 g, 7.63 mmol) and NaHCO3 (1.28 g, 15.3 mmol) were suspended in DMF (10 ml). Methyl iodide (2.37 ml, 5.41 g, 38.13 mmol) was added to the mixture, which was then heated at 500C overnight. After concentrating the reaction in vacuo, the residue was partitioned between ethyl acetate and water. The aqueous layer was extracted once more with ethyl acetate, and the combined organic layers were washed with brine, dried (MgSO4) and concentrated. The residue was purified by silica flash chromatography (gradient elution, using 1 : 1 hexane-ethyl acetate and ethyl acetate) to provide the title compound as a colorless oil (1.66 g, 89%).
[00556] Step 2: (2R.3R)-tert-butyl 3-hvdroxy-2-(hvdroxymethyl)pyrrolidine-l- carboxylate
Figure imgf000126_0003
[00557] To a stirring solution of the (2S,3R)-l-(tert-butoxycarbonyl)-3- hydroxypyrrolidine-2-carboxylic acid (0.36 g, 1.47 mmol) in THF (5 ml) was added LiCl (0.19 g, 4.4 mmol) followed by NaBH4 (0.17 g, 4.4 mmol). After addition of ethanol (10 ml), the resulting mixture was stirred at room temperature overnight. The reaction flask was placed in an ice bath, and the cooled milky white solution was acidified to pH 2-3 with 37% HCl. The solution was concentrated, and the residue was partitioned between ethyl acetate and water. The aqueous layer was extracted with ethyl acetate, and the combined organic layers were washed with brine, dried (MgSO4) and concentrated. Purification of the resulting oil by flash chromatography (silica gel, using ethyl acetate) provided 0.30 g (95%) of the title compound as a colorless oil.
[00558] Step 3: (2R.3R)-2-(hvdroxymethyl)pyrrolidin-3-ol hydrochloride [00559] 4N HCl/dioxane solution (5 ml) was added to (2R,3R)-tert-butyl 3-hydroxy-2-
(hydroxymethyl)pyrrolidine-l-carboxylate and the mixture was stirred for 2 hours. The solution was concentrated in vacuo. The residue was diluted with toluene (5 ml) and reconcentrated to provide the title compound as a colorless oil (0.21 g, 100%). [00560] (3R.5R)-5-(Hvdroxymethyl)pyrrolidin-3-ol hydrochloride
Figure imgf000127_0001
[00561] Step 1 : (2R,4R)-1 -((benzyl oxy)carbonyl)-4-hvdroxypyiroridine-2-carboxyric acid
Figure imgf000127_0002
[00562] cis-4-Hydroxy-D-proline (1.0 g, 7.63 mmol) and NaHCO3 (1.6 g, 19.05 mmol) were dissolved in H2O (16 ml), then a solution of benzyl chloroformate (1.25 ml, 1.49 g, 8.76 mmol) in toluene (4 ml) was added dropwise over a period of 15 minutes. After stirring at room temperature for 16 hours, the two phases were separated. Excess benzyl chloroformate was removed from the aqueous phase by washing with ether (4x5 ml). Acidification of the aqueous phase to pH 2 with concentrated HCl caused the oily product to precipitate and this was extracted into ethyl acetate by repeated washings (3x5 ml) of the aqueous layer. The combined organic layers were dried (MgSO4) and concentrated to give the title compound as a viscous oil (2.02 g, 100%).
[00563] Step 2: (2R, 4R)-I -Benzyl 2 -methyl 4-hvdroxypyrrolidine-L2-dicarboxylate
HQ
-N/" 1CO2Me CO2Bn
[00564] The (2R,4R)-1 -((benzyl oxy)carbonyl)-4-hydroxypyrrolidine -2 -carboxylic acid
(2.02 g, 7.63 mmol) and NaHCO3 (1.28 g, 15.3 mmol) were suspended in DMF (10 ml). Methyl iodide (2.37 ml, 5.41 g, 38.13 mmol) was added to the mixture, which was then stirred and heated at 500C overnight. After concentrating the reaction under reduced pressure, the residue was partitioned between ethyl acetate and water. The aqueous layer was extracted once more with ethyl acetate, and the combined organic layers were washed with brine, dried (MgSO4) and concentrated. The residue was purified by silica flash chromatography (gradient elution, using 1 : 1 hexane-ethyl acetate and ethyl acetate) to provide the title compound as a colorless oil (1.9 g, 89%).
[00565] Step 3: (2R.4RVBenzyl 4-hvdroxy-2-(livdroxymethvnpyrrolidine-l- carboxylate
Figure imgf000128_0001
[00566] To a stirring solution of the (2R, 4R)-I -benzyl 2-methyl 4-hydroxypyrrolidine-
1,2-dicarboxylate (0.41 g, 1.47 mmol) in THF (5 ml) was added LiCl (0.19 g, 4.4 mmol) followed by NaBH4 (0.17 g, 4.4 mmol). After addition of ethanol (10 ml), the resulting mixture was stirred at room temperature overnight. The reaction flask was placed in an ice bath, and the cooled milky white solution was acidified to pH 2-3 with 37% HCl. The solution was concentrated, and the residue was partitioned between ethyl acetate and water. The aqueous layer was extracted with ethyl acetate, and the combined organic layers were washed with brine, dried (MgSO4) and concentrated. Purification of the resulting oil by flash chromatography (silica gel, using ethyl acetate) provided the title compound (0.35 g, 95%) as a colorless oil.
[00567] Step 4: (3R.5R)-5-(hvdroxymethyl)pyrrolidin-3-ol hydrochloride
[00568] (2R,4R)-benzyl 4-hydroxy-2-(hydroxymethyl)pyrrolidine-l-carboxylate (0.35 g, 1.4 mmol) was dissolved in ethanol (30 ml) and transferred into a Parr shaker bottle. After adding 10% Pd-C (0.07 g), the mixture was shaken under an atmosphere of hydrogen at 50 psi for 0.5 h on the Parr apparatus. The catalyst was removed by filtration through Celite. The filter cake was washed with ethanol and the combined filtrate and washings were concentrated in vacuo to yield a colorless oil. For ease of handling, the amine was converted to the hydrochloride salt. A 4N HCl/dioxane solution (1 ml) was added to the residue, along with enough methanol (~1 ml) to completely dissolve the residue. After complete mixture, the solvent was evaporated under reduced pressure. The solid was diluted with toluene (20 ml) and reconcentrated. Finally, the solid was triturated with ether, the ether was discarded, and the solid was dried in vacuo to yield 0.186 g (87%) of the title compound as a pink solid. [00569] (3R. 5S)-5-(Hvdroxymethyl)pyrrolidin-3-ol hydrochloride
HO,
OH
H HCI [00570] To N-Boc-trans-4-hydroxy-L-prolinol (0.422 g, 1.94 mmol) was added a 4N
HCl/dioxane solution (5 ml) and the mixture was swirled for 1 hour. The solution was concentrated in vacuo. The residue was diluted with toluene (10 ml) and reconcentrated. The resulting white solid was triturated with ethyl ether, the ether was discarded, and the solid dried under vacuum to provide 0.29 g (97%) of the title compound as a colorless white solid. 1H NMR (D2O 400 MHz) 4.65-4.67 (m, IH), 3.99-4.06 (m, IH), 3.93 (dd, J=12.5Hz, 3.6 Hz, IH), 3.71 (dd, J=12.5 Hz, 6.9 Hz, IH), 3.44 (dd, J=12.7 Hz, 3.8 Hz, IH), 3.32 (d, J=12.7 Hz, IH), 2.11-2.17 (m, IH), 1.92-1.98 (m, IH).
[00571] tert-Butyl (3R,5R)-5-(hydroxymethyl)pyrrolidin-3-ylcarbamate
Figure imgf000129_0001
[00572] Step 1 : (2R, 4S)-I -benzyl 2 -methyl 4-hvdroxypyrrolidine-l,2-dicarboxylate
Figure imgf000129_0002
[00573] To a stirred solution of the (2R, 4R)-I -benzyl 2-methyl 4-hydroxypyrrolidine-
1,2-dicarboxylate (1.45 g, 5.2 mmol), triphenylphosphine (4.59 g, 17.5 mmol), and p- nitrobenzoic acid (2.6 g, 15.6 mmol) in dry benzene (10 ml) at room temperature was added diethylazodicarboxylate dropwise (2.57 ml, 17.5 mmol). The solution was then stirred at room temperature for 6 h, whereupon the volatile components were removed in vacuo and the residue purified by flash chromatography (silica gel, hexane-ethyl ether 1: 1, and again with hexane-ethyl ether-methylene chloride 2: 1 : 1). This residue was dissolved in methanol (10 ml), K2CO3 was added (0.02 g, 0.14 mmol) and the mixture was stirred for 1 h at room temperature. After removing the volatile components in vacuo, the residue was purified by flash chromatography (silica gel, gradient elution, using ethyl ether to ethyl acetate) to provide the title compound as a colorless oil (0.33 g, 23%).
[00574] Step 2 : (3 S.5R)- 1 -((Benzyloxy)carbonyl)-5-(methoxycarbonyl)pyrrolidin-3 -yl
4-methylbenzenesulfonate
Figure imgf000129_0003
[00575] (2R,4S)-1 -benzyl 2-methyl 4-hydroxypyrrolidine-l,2-dicarboxylate (0.33 g, 1.18 mmol) and DMAP (0.43 g, 3.55 mmol) were dissolve in chloroform and the mixture cooled to -5°C in an ice-ethanol bath. p-Toluenesulfonyl chloride (0.45 g, 2.24 mmol) was added and the reaction was stirred while warming to room temperature for 2 hours. After quenching with water (0.6 ml) and vigorous stirring for 10 minutes, the layers were separated and the aqueous layer extracted with methylene chloride (2x). The organic layers were dried (MgSO4) filtered through a plug of silica gel (7 ml packed in ethyl ether), eluted with ethyl ether and concentrated. The residue was purified by flash chromatography (gradient elution, using hexane-ethyl ether 1 : 1 to ethyl ether) to provide the title compound as a colorless oil (0.49 g, 95%).
[00576] Step 3: (2R,4R)-1 -benzyl 2-methyl 4-azidopyrroridine-l,2-dicarboxylate
Ns
^N' '11CO2Me
CO2Bn
[00577] Sodium azide (0.33 g, 5.07 mmol) was added to the (3S,5R)-1-
((benzyloxy)carbonyl)-5-(methoxycarbonyl)pyrrolidin-3-yl 4-methylbenzenesulfonate (0.49 g, 1.13 mmol) in DMF (8 ml) and the mixture heated at 500C overnight. After concentrating in vacuo, the residue was partitioned between ethyl ether and water. The aqueous layer was extracted with ethyl ether, and the combined organic layers were dried (MgSO4) and concentrated. Purification of the resulting oil by flash chromatography (silica gel, using ethyl ether) provided 0.33 g (97%) of the title compound as a colorless oil. [00578] Step 4: (3R.5R)-l-((benzyloxy)carbonyl)-5-(methoxycarbonyl)pyrrolidin-3- ylcarbamate
Figure imgf000130_0001
[00579] Triphenylphosphine (0.33 g, 1.15 mmol) was added to a solution of the
(2R,4R)-1 -benzyl 2-methyl 4-azidopyrrolidine-l,2-dicarboxylate (0.33 g, 1.08 mmol) in THF (4 ml) and water (2 ml). After stirring at 50 0C overnight, sodium bicarbonate (0.23 g, 2.71 mmol) was added, followed by di-tert-butyl dicarbonate (0.47 g, 2.17 mmol) and stirring was continued at 500C for another 4 h. Volatiles were removed under reduced pressure, and the residue was partitioned between ethyl ether and water. The aqueous layer was extracted with ethyl ether, and the combined organic layers were dried (MgSO4) and concentrated. Purification of the resulting oil by silica gel flash chromatography (gradient elution, using hexane-ethyl ether 1: 1 to 3:7) provided 0.258 g (64%) of the title compound as a colorless oil. [00580] Step 5: tert-butyl (3R.5R)-l-((benzyloxy)carbonyl)-5-
(hydroxymethyl)pyrrolidin-3-ylcarbamate
Figure imgf000131_0001
[00581] To a stirring solution of the tert-butyl (3R,5R)-l-((benzyloxy)carbonyl)-5-
(methoxycarbonyl)pyrrolidin-3-ylcarbamate (0.16 g, 0.42 mmol) in THF (1.5 ml) was added LiCl (0.054 g, 1.27 mmol) and NaBH4 (0.048 g, 1.27 mmol). After addition of ethanol (3 ml), the resulting mixture was stirred at room temperature overnight, and then quenched with water (1 ml). The solution was concentrated, and the residue was partitioned between ethyl acetate (20 ml) and water (3 ml). The aqueous layer was extracted with ethyl acetate (2x 2ml), and the combined organic layers were washed with brine, dried (MgSO4) and concentrated. Purification of the resulting oil by flash chromatography (silica gel, using ethyl ether) provided 0.11 g (74%) of the title compound as a colorless oil. [00582] Step 6: tert-butyl GR.5R)-5-(hvdroxymethvnpyrrolidin-3-ylcarbamate
[00583] tert-Butyl (3R,5R)-l-((benzyloxy)carbonyl)-5-(hydroxymethyl)pyrrolidin-3- ylcarbamate (0. H g, 0.31 mmol) was dissolved in ethanol (20 ml) and transferred to a Parr shaker bottle. After adding 10% Pd-C (0.030 g), the mixture was shaken under an atmosphere of hydrogen at 50 psi for 0.5 h on the Parr apparatus. The catalyst was removed by filtration through Celite. The filter cake was washed with ethanol and the combined filtrate and washings were concentrated in vacuo to yield the title compound as a colorless oil (0.07 g, 100%). [00584] (2R.3 S)-2-(Hvdroxymethvnpyrrolidin-3-ol hydrochloride
.OH
Q H H-Cl '
[00585] Step 1 : (2S.3S)-l-(tert-butoxycarbonyl)-3-hydroxypyrrolidine-2-carboxylic acid
.-OH
Figure imgf000131_0002
[00586] Trans-3-hydroxy-L-proline (2.62 g, 20.0 mmol) and sodium bicarbonate (5.04 g, 60 mmol) were dissolved in water (20 ml). Dioxane was added (20 ml) followed by di- tert-butyl-dicarbonate (8.72 g, 40 mmol). Stirring was continued at room temperature overnight. The reaction was concentrated, and the residue was partitioned between ethyl ether (10 ml) and water (30 ml). The aqueous layer was washed once more with ether, and the organic layers were discarded. Gradual acidification of the aqueous phase with concentrated HCl caused the oily product to precipitate and this was extracted into ethyl acetate by repeated washings (3x10 ml) of the aqueous layer. The combined organic layers were washed with brine, dried (MgSO4) and concentrated to provide the title compound as a viscous oil (4.17 g, 90%).
[00587] Step 2: (2S.3 S)- l-tert-Butyl-2 -methyl 3-hydroxypyrrolidine-1.2-dicarboxylate
PH
CX CO2Me
Boc
[00588] The (2S,3S)-l-(tert-butoxycarbonyl)-3-hydroxypyrrolidine-2-carboxylic acid
(4.2 g, 18.2 mmol) and NaHCO3 (3.1 g, 36.3 mmol) were suspended in DMF (20 ml). Methyl iodide (5.7 ml, 12.9 g, 91.0 mmol) was added to the mixture, which was then heated at 500C overnight. After concentrating the reaction in vacuo, the residue was partitioned between ethyl acetate and water. The aqueous layer was extracted once more with ethyl acetate, and the combined organic layers were washed with brine, dried (MgSO4) and concentrated. The residue was purified by silica flash chromatography (gradient elution, using 1 : 1 hexane-ethyl ether to ethyl ether) to provide the title compound as a colorless oil (3.7 g, 82%).
[00589] Step 3: (2R.3S)-tert-butyl 3-hvdroxy-2-(hvdroxymethyl)pyrrolidine-l- carboxylate
Figure imgf000132_0001
[00590] The (2S,3S)-l-tert-butyl-2-methyl 3-hydroxypyrrolidine-l,2-dicarboxylate
(0.54g, 2.20 mmol) was dissolved in THF (8.0 ml). Lithium chloride (0.28 g, 6.60 mmol) and sodium borohydride (0.25 g, 6.60 mmol) were added, then ethanol (16.0 ml). The reaction was stirred overnight, then quenched with water (4 mL) and concentrated. The residue was partitioned between ethyl acetate and water. The aqueous layer was extracted with ethyl acetate. The organic layers were combined, washed with brine, and dried (MgSO4). Flash chromatography (50 ml silica gel, gradient elution, using ethyl acetate to 9: 1 ethyl acetate- ethanol) yielded 0.5 grams (100%) of the title compound as a colorless solid. [00591] Step 4: (2R.3S)-2-(hvdroxymethvnpyrrolidin-3-ol hydrochloride
[00592] To (2R,3S)-tert-butyl 3-hydroxy-2-(hydroxymethyl)pyrrolidine-l-carboxylate
(0.50 g, 2.30 mmol) was added a 4N HCl/dioxane solution (6 ml) and the mixture was swirled for 2 hours. The solution was concentrated in vacuo. The residue was diluted with toluene (20 ml) and reconcentrated to provide the title compound as a colorless oil (0.36 g, 100%).
[00593] (2R.3R.4S)-2-(Hvdroxymethyl)pyrrolidine-3.4-diol hydrochloride
HO OH
.OH
H HCl
[00594] Step 1 : (S)-l-(tert-butoxycarbonyl)-2,5-dihydro-lH-pyrrole-2-carboxylic acid
Figure imgf000133_0001
[00595] 3,4-Dehydro-L-Proline (1.0 g, 8.8 mmol) was dissolved in H2O (9.0 ml) and sodium bicarbonate (2.23 g, 26.5 mmol). Dioxane (9.0 ml) was added followed by di-tert- butyldicarbonate (3.86 g, 17.7 mmol). The reaction was stirred overnight, then concentrated. The residue was partitioned between ethyl ether (20 ml) and water (25 ml), and the layers were separated. The aqueous layer was diluted with ethyl acetate (20 ml) and the mixture was slowly acidified with concentrated HCl while stirring the mixture vigorously to extract the precipitate into the organic layer. After acidification to ~ pH2, and further extraction with ethyl acetate, the aqueous layer was saturated with salt and extracted once more with ethyl acetate. The combined organic layers were washed with brine, dried (MgSO4) and concentrated to yield the title compound (2.0 g, 100%) as a viscous, colorless oil. [00596] Step 2: (S)-l-tert-butyl-2-methyl 2H-pyrrole-1.2(5H)-dicarboxylate
\N/""*CO2Me
Boc
[00597] The (S)-l-(tert-butoxycarbonyl)-2,5-dihydro-lH-pyrrole-2-carboxylic acid
(0.85 g, 4.0 mmol) was dissolved in ethyl ether (10 ml) and methanol (10 ml), then cooled to -5°C in an ice/ethanol bath. Trimethylsilyldiazomethane (4.4 ml of 2.0M solution in hexane, 8.8 mmol) was added dropwise. After stirring overnight, volatiles were removed under reduced pressure. The residue was partitioned between ethyl ether (20 ml) and water (5 ml), and the layers were separated. The organic layer was washed with saturated NaHCθ3 and brine, then dried (MgSO4), filtered through a silica gel plug (7 ml) with ethyl ether, and concentrated, yielding the title compound as a colorless oil (0.821 g, 91%).
[00598] Step 3: (2S.3R.4S)-l-tert-Butyl 2-methyl 3.4-dihydroxypyrrolidine-1.2- dicarboxylate
HQ. PH
Q- CO2Me
Boc
[00599] (S)-l-tert-Butyl-2-methyl 2H-pyrrole-l,2(5H)-dicarboxylate (0.83 g, 3.65 mmol) was dissolved in tert-butyl alcohol (15 ml), tetrahydrofuran (4 ml), and water (1.3 ml). Osmium tetraoxide (0.37 ml of 100 mg/ml solution in tert-butyl alcohol, 0.15 mmol) was added, followed by N-methylmorpholine N-oxide (0.51 g, 4.4 mmol). The reaction was stirred at room temperature for 5 hours, and then diluted with saturated sodium thiosulfate (5 ml), ethyl acetate (15 ml) and water (5 ml). After separation of layers, the organic layer was washed once more with sodium thiosulfate, then with brine, dried (MgSO4), and filtered through a silica gel plug (7 ml) with ethyl acetate (75 ml) and concentrated. The oil was taken up in minimal ethyl ether/methylene chloride and purified by flash chromatography (gradient elution, using hexane-ethyl acetate 3:7 to ethyl acetate) to provide the title compound as a colorless oil (0.83 g, 87%).
[00600] Step 4: 3aR.4S.6aSV5-tert-Butyl 4-methyl tetrahvdro-2.2-dimethyl-
|"1.3"ldioxolo|"4.5-c"lpyrrole-4.5-dicarboxylate
CX CO2Me
Boc
[00601] The (2S,3R,4S)- 1 -tert-butyl 2-methyl 3,4-dihydroxypyrrolidine- 1 ,2- dicarboxylate (0.426 g, 1.63 mmol) was dissolved in 2,2-dimethoxypropane (10 ml). Pyridinium p-toluenesulfonate (0.02 g, 0.08 mmol) was added and the reaction was stirred at room temperature overnight. TLC showed reaction to be almost complete. More 2,2- dimethoxypropane (5 ml) was added and the mixture was heated with a heat gun until the mixture boiled and the total volume was reduced by about 1/4 (took about 5 minutes). The reaction was diluted with ethyl ether (10 ml) and the solution was extracted with saturated NaHCO3 and brine, then it was dried (MgSO4), filtered and concentrated, providing a pale yellow oil. Flash chromatography (70 ml silica gel, gradient elution, using hexane-ethyl ether 25: 15 to hexane-ethyl ether 1: 1) yielded the title compound (0.402 g, 82%) as colorless oil. [00602] Step 5: (3aR.4R.6aS)-tert-butyl tetrahydro-4-(hydroxymethyl)-2.2-dimethyl- ri,3"|dioxolor4,5-c"|pyrrole-5-carboxylate
Figure imgf000135_0001
[00603] The (3aR,4S,6aS)-5-tert-butyl 4-methyl tetrahydro-2,2-dimethyl-
[l,3]dioxolo[4,5-c]pyrrole-4,5-dicarboxylate (0.40 g, 1.3 mmol) was dissolved in THF (5.0 ml) and treated sequentially with lithium chloride (0.17 g, 4.0 mmol), sodium borohydride (0.15 g, 4.0 mmol) and ethanol (10 ml). The reaction was stirred overnight, then quenched with water (3 mL) and concentrated. The residue was partitioned between ethyl acetate and water. After extracting the aqueous layer once more with ethyl acetate, the combined organic layers were washed with brine and dried (MgSO4). Flash chromatography (50 ml silica gel, gradient elution, using hexane-ethyl ether 6:4 to ethyl ether) yielded 0.36 grams (99%) of the title compound as a colorless oil.
[00604] Step 6: (2R.3R.4S)-2-(hvdroxymethyl)pyrrolidine-3.4-diol hydrochloride
[00605] The (3aR,4R,6aS)-tert-butyl tetrahydro-4-(hydroxymethyl)-2,2-dimethyl-
[l,3]dioxolo[4,5-c]pyrrole-5-carboxylate (0.36 g, 1.3 mmol) was dissolved in 4N HCl/dioxane (5 mL) and water (0.5 ml) and the reaction was swirled at room temperature for 2 hours. Then volatiles were removed under reduced pressure to yield a pink oil. This residue was diluted with toluene (20 mL) and reconcentrated to a solid which was triturated with ethyl ether. The ether was discarded and the solid dried in vacuo. Yield was 200 mg (90%) of the title compound as a pink solid. 1H NMR (D2O 400 MHz) 4.37-4.39 (m, IH), 4.21(dd, J=8.6 Hz, 4.1 Hz, IH), 3.98 (dd, J=12.7 Hz, 3.5 Hz, IH), 3.83 (dd, J=12.5Hz, 6.0 Hz, IH), 3.62 (ddd, J = 8.6 Hz, 6.0 Hz, 3.5 Hz, IH), 3.50 (dd, J = 13.0 Hz, 4.1 Hz, IH), 3.37 (dd, J=13.0 Hz, 2.0 Hz, IH). [00606] (2R.3S.4S)-2-(Hvdroxymethvnpyrrolidine-3.4-diol hydrochloride
Figure imgf000135_0002
[00607] Step 1 :
Figure imgf000136_0001
[00608] The 2,3,5-tri-O-benzyl-β-L-arabinose (0.5g, 1.19 mmol) was dissolved in ethanol (5 ml) with heating. Sodium bicarbonate (249 mg, 2.96 mmol) in water (2.5 ml) was added, followed by hydroxylamine hydrochloride (247 mg, 3.55 mmol). The heterogeneous mixture was stirred at room temperature for 5 hours. Then more hydroxylamine hydrochloride (100 mg, 1.44 mmol) and sodium bicarbonate (100 mg, 1.19 mmol) was added and the reaction stirred overnight. More sodium bicarbonate (0.084 g, lmmol) was added and the mixture was heated to boiling for 5 minutes. After cooling to room temperature, the reaction was concentrated. The resulting oil was triturated with THF (20 ml) until solids were a fine powder. The solids were filtered off and the filtrate was concentrated. The residue was purified by flash chromatography (hexane:ethyl acetate 3: 1) to yield 0.45 grams (87%) of the product as a colorless oil. [00609] Step 2:
Figure imgf000136_0002
[00610] A solution of the oxime of 2,3,5-tri-O-benzyl-β-L-arabinose (0.45 g, 1.0 mmol) in dry ethyl ether (5 ml) was added dropwise to a solution Of LiAlH4 (0.75 mL of 2.5 M in THF, 1.85 mmol). The mixture was stirred for an additional 2 hours at room temperature after the addition. Ethyl acetate (1.7 ml) was added slowly to decompose the excess LiAlH4, followed by 0.75 mL of 4N NaOH solution. The resulting cloudy suspension was filtered through a bed of Celite, and the Celite cake was washed thoroughly with ether and ethyl acetate. The filtrate and washings were concentrated to provide a viscous oil. The oil was taken up in ethyl acetate (20 ml) and washed with saturated NaHCθ3 and brine, then dried (MgSO4) and concentrated to provide the crude amine as a pale yellow oil (0.44 g, 100%). This amine (0.44 g, 1.0 mmol) was dissolved in THF (3 ml) and water (1.5 ml). Sodium bicarbonate (0.22 g, 2.6 mmol) was added followed by di-tert-butyldicarbonate (0.46 g, 2.1 mmol). The reaction was stirred overnight, then concentrated. The residue was partitioned between ethyl ether (20 ml) and water (10 ml), and the layers were separated. The aqueous layer was extracted once more with ethyl ether, and the combined organic layers were washed with brine, dried (MgSO4) and concentrated. Flash chromatography (silica gel, hexane-ethyl ether 1: 1) yielded 0.29 g (52%) of the Boc amine as a colorless oil. [00611] Step 3:
Figure imgf000137_0001
[00612] The alcohol (0.29 g, 0.55 mmol) and triethylamine (0.13 mL, 0.96 mmol) were dissolved in methylene chloride (2.0 ml) and cooled to -5°C in an ice-ethanol bath. Methanesulfonyl choride (64 μL, 0.83 mmol) was added and the reaction was stirred for 2 hours. After quenching with water (0.2 ml) the mixture was stirred for 30 minutes. The layers were separated, the aqueous layer was washed with methylene chloride and the combined organic layers were dried (MgSO4) and concentrated. The residue was purified by flash chromatography (hexane-ethyl ether-methylene chloride 2: 1 : 1) to yield the title compound (0.30 g, 91%).
[00613] Step 4:
Figure imgf000137_0002
[00614] The N-Boc-O-mesylate compound (0.247 g, 0.412 mmol) was dissolved in
DMF (2.0 mL), then sodium hydride (0.023 grams of 60% oil dispersion) was added directly to the solution and the cloudy mixture was stirred at room temperature for 2.5 hours. TLC showed the reaction to be complete. The reaction was diluted with ethyl ether (8 ml) and filtered directly through a plug of silica gel (7 ml packed in ethyl ether), and the filter cake was washed with ether. After removing volatiles under reduced pressure, the resulting residue was purified by silica gel flash chromatography, (hexane-ethyl ether-methylene chloride 3: 1 : 1) to yield 0.196 g (95%) of the pyrrolidine product as colorless oil. [00615] Step 5: (2R.3 S.4S)-tert-Butyl 3.4-dihydroxy-2-(hvdroxymethyl) pyrrolidine- 1 - carboxylate
Figure imgf000138_0001
[00616] The tri-O-benzyl pyrrolidine (0.24 g, 0.47 mmol) was dissolved in methanol
(50 ml) and added to a Parr shaker bottle. After flushing with nitrogen, 10% Pd-C (150 mg) was added and the mixture was hydrogenated on the Parr apparatus at 50 psi H2 for 4 hours. The reaction mixture was filtered through a pad of Celite, the filter cake washed with methanol, and the solution was concentrated. Purification of the resulting residue by flash chromatography (silica gel, gradient elution, using ethyl acetate to ethyl acetate-ethanol 9: 1) gave the triol product (0.103 g, 95%) as a colorless oil.
[00617] Step 6: (2R.3S.4S)-2-(Hvdroxymethvnpyrrolidine-3.4-diol hydrochloride
Figure imgf000138_0002
[00618] The N-B oc -pyrrolidine (0.103 g, 0.442 mmol) was dissolved in 4N
HCl/dioxane (3 mL) and swirled at room temperature for 1.5 hour. The reaction was concentrated in vacuo to yield a colorless oil. The residue was diluted with toluene (20 mL), reconcentrated, then triturated with ethyl ether to try to induce crystallization. The residue solidified, the ether was discarded, and the solid was dried under vacuum to provide the crude title compound as a white solid (75 mg, 100%). 1H NMR (D2O 400 MHz) 4.39 (d, J = 4.3 Hz, IH), 4.32 (app s, IH), 3.99-4.05 (m, IH), 3.87-3.93 (m, 2H), 3.66 (dd, J = 13.0 Hz, 4.3 Hz, IH), 3.30 (d, J=13.0 Hz, IH).
[00619] EXAMPLE 5; 3-(2-Fluoro-4-iodo-phenylamino)-furo[3.2-c1pyridine-2- carboxylic acid ((R)-2,3-dihydroxy-propoxy)-amide
Figure imgf000138_0003
[00620] Step 1 : 3-(2-Fluoro-4-iodo-phenylamino)-furor3.2-clpyridine-2-carboxylic acid ((R)-2 ,2-dimethyl- [1,31 dioxolan-4-ylmethoxy)-amide
Figure imgf000139_0001
[00621] A mixture of ethyl 3-(2-fluoro-4-iodo-phenylamino)-furo[3,2-c]pyridine-2- carboxylate (60 mg, 0.15 mmol), IN aqueous sodium hydroxide solution (0.18 ml, 0.18 mmol) and methanol (2 ml) were heated at 650C for 30 minutes. The reaction mixture was concentrated then azeotroped with toluene (3 x 10 ml) to give a solid residue. The solid residue was dissolved in anhydrous THF (2 ml) and O-((R)-2,2-dimethyl-[l,3]dioxolan-4- ylmethyl)hydroxylamine (35 mg, 0.23 mmol), EDCI (28 mg, 0.15 mmol), HOBt (22 mg, 0.16 mmol) and DIPEA (61 μl, 0.35 mmol) were added. After stirring overnight at 4O0C, the residue was absorbed onto HM-N and purified by flash chromatography (Si-SPE, diethyl ether: MeOH, gradient 100:0 to 90: 10) to afford the title compound as a yellow foam (30 mg, 48%). LCMS (method B): Rx = 3.10 min, M+H+ = 528.
[00622] Step 2: 3-(2-Fluoro-4-iodo-phenylamino)-furor3,2-c1pyridine-2-carboxylic acid ((R)-2,3-dihydroxy-propoxy)-amide
[00623] 3-(2-Fluoro-4-iodo-phenylamino)-furo[3,2-c]pyridine-2-carboxylic acid ((R)-
2,2-dimethyl-[l,3]dioxolan-4-ylmethoxy)-amide (30 mg, 0.06 mmol) was dissolved in methanol (0.5 ml) and loaded onto an Isolute® SCX-2 cartridge (5g). The cartridge was then washed with methanol (15 ml) and the desired product was subsequently eluted using2M NH3 in MeOH and the eluant was collected and concentrated to give a residue. The residue was absorbed onto HM-N and purified by flash chromatography (Si-SPE, diethyl ether: MeOH, gradient 100:0 to 80:20) to afford the title compound as a white solid (18 mg, 64%). LCMS (method A): Rx = 5.80 min, M+H+ = 488. IH NMR (d4-MeOH, 400MHz) 8.53 (d, J = 5.9 Hz, IH), 8.49 (d, J = 1.0 Hz, IH), 7.62 (dd, J = 5.9 Hz, 1.0 Hz, IH), 7.60 (dd, J = 10.3 Hz, 2.0 Hz, IH), 7.51 (dd, J = 8.5 Hz, 2.0 Hz, IH), 7.06 (t, J = 8.5 Hz, 1.0 Hz, IH), 4.10 (m, IH), 3.96 (m, 2H), 3.63 (m, 2H).
[00624] EXAMPLE 6; 3-(2-Fluoro-4-bromo-phenylamino)-furor3.2-c1pyridine-2- carboxylic acid ((R)-2.3-dihydroxy-propoxy)-amide
Figure imgf000139_0002
[00625] Step 1 : 3-(2-Fluoro-4-bromo-phenylamino)-furor3.2-clpyridine-2-carboxylic acid ((R)-2 ,2-dimethyl- [1,31 dioxolan-4-ylmethoxy)-amide
Figure imgf000140_0001
[00626] A mixture of ethyl 3-(2-fluoro-4-bromo-phenylamino)-furo[3,2-c]pyridine-2- carboxylate (400 mg, 1.06 mmol), IN aqueous sodium hydroxide solution (1.11 ml, 1.11 mmol) and methanol (10 ml) were heated at 650C for 30 minutes. The reaction mixture was concentrated in vacuo then azeotroped with toluene (3 x 10 ml) to give a solid residue. The solid residue was dissolved in anhydrous THF (10 ml) and O-((R)-2,2-dimethyl- [l,3]dioxolan-4-ylmethyl)hydroxylamine (255 mg, 2.12 mmol), EDCI (254 mg, 1.32 mmol), HOBt (200 mg, 1.48 mmol) and DIPEA (556 μl, 3.18 mmol) were added. After stirring overnight at ambient temperature, the reaction mixture was concentrated in vacuo to afford a yellow residue. The resultant residue was dissolved in ethyl acetate (30 ml), washed with water (30 ml) followed by brine (30 ml) before the organic layer was isolated then dried over sodium sulfate and concentrated in vacuo to afford a yellow oil. The oil was purified by flash chromatography (Si-SPE, pentane: ethyl acetate, gradient 50:50 to 0: 100) to afford the title compound as an off-white foam (370 mg, 73%). IH NMR (CDCl3, 400 MHz) 9.22 (s, IH), 7.95 (s, IH), 7.67 (m, IH), 7.51 (m, IH), 7.45-7.31 (m, 3H), 7.13 (t, J = 8.4 Hz, IH), 4.49 (m, IH), 4.08-4.26 (m, 3H), 3.89 (m, IH), 1.49 (s, 3H), 1.40 (s, 3H).
[00627] Step 2: 3-(2-Fluoro-4-bromo-phenylamino)-furor3,2-c1pyridine-2-carboxylic acid ((R)-2.3-dihydroxy-propoxy)-amide
[00628] To 3-(2-Fluoro-4-bromo-phenylamino)-furo[3,2-c]pyridine-2-carboxylic acid
((R)-2,2-dimethyl-[l,3]dioxolan-4-ylmethoxy)-amide (50 mg, 0.10 mmol) was added a solution of 4N HCl in methanol (1 ml) then the reaction mixture was stirred at ambient temperature for 30 minutes. Water (10 ml) and ethyl acetate (10 ml) were added to the reaction mixture and the organic layer was isolated. The resultant organic phase was washed with saturated NaHCO3 solution (10 ml) then dried over sodium sulfate before being concentrated in vacuo to produce a residue. The residue was loaded onto an Isolute® SCX-2 cartridge (5g). The cartridge was then washed with methanol (15 ml) before the desired product was eluted using 2M ammonia in MeOH and the eluant was collected then concentrated to give a residue. The residue was purified by flash chromatography (Si-SPE, DCM: MeOH, gradient 100:0 to 93:7) to afford the title compound as a white solid (27 mg, 59%). LCMS (method A): Rx = 5.55 min, M+H+ = 440/442. IH NMR (d4-MeOH, 400MHz) 8.52 (s, IH), 8.44 (s, IH), 7.60 (d, J = 5.9 Hz, IH), 7.44 (dd, J = 8.8 Hz, 2.2 Hz, IH), 7.32 (m, IH), 7.19 (m, IH), 4.06 (m, IH), 3.91 (m, 2H), 3.59 (m, 2H).
[00629] EXAMPLE 7; 3-(4-Ethvnyl-2-fluoro-phenylamino)-furor3.2-clpyridine-2- carboxylic acid (YR)-2.3-dihvdroxy-propoxy)-amide
Figure imgf000141_0001
[00630] A mixture of 3-(2-fluoro-4-bromo-phenylamino)-furo[3,2-c]pyridine-2- carboxylic acid ((R)-2,2-dimethyl-[l,3]dioxolan-4-ylmethoxy)-amide (100 mg, 0.21 mmol), trimethylsilyl acetylene (288 μl, 2.08 mmol) and PdCl2(PPh3)2 (7.3 mg, 0.01 mmol) in triethylamine (3.0 ml) were subjected to microwave irradiation at 15O0C for 10 minutes. The reaction mixture was then diluted with ethyl acetate (5 ml) and the resultant solution was washed with water (10 ml) followed by brine (5 ml) then the organic layer was dried over sodium sulfate and concentrated in vacuo to give a residue. The residue was dissolved in methanol (3 ml) and potassium carbonate (58 mg, 0.42 mmol) was added, and the reaction mixture was stirred for 1 hour at ambient temperature. The reaction mixture was then evaporated to dryness and the resultant residue dissolved in ethyl acetate (20 ml). The organic phase was washed with water (10 ml) followed by brine (10 ml) and dried over sodium sulfate the concentrated in vacuo to produce a residue. The resultant residue was dissolved in methanol (0.5 ml) and loaded onto an Isolute® SCX-2 cartridge (5g). The cartridge was washed with methanol (15 ml) before the desired product was eluted using 2M triethylamine in MeOH, and the eluant was collected then concentrated to afford the title compound as a white solid (19 mg, 24%). LCMS (method A): Rx = 5.78 min, M+H+ = 386. IH NMR (d4- MeOH, 400MHz) 8.51 (m, 2H), 7.60 (d, J = 5.9 Hz, IH), 7.31 (dd, J = 11.3 Hz, 1.8 Hz, IH), 7.23 (dd, J = 8.4 Hz, 1.6 Hz, IH), 7.06 (t, J = 8.5 Hz, IH), 4.07 (m, IH), 3.92 (m, 2H), 3.55 (m, 2H), 3.49 (s, IH).
[00631] EXAMPLE 8: 3-(2-Fluoro-4-iodo-phenylamino)-furor3.2-c1pyridine-2- carboxylic acid cvclopropylmethoxy-amide
Figure imgf000141_0002
A mixture of ethyl 3-(2-fluoro-4-iodo-phenylamino)-furo[3,2-c]pyridine-2-carboxylate (87 mg, 0.20 mmol), IN aqueous sodium hydroxide solution (0.21 ml, 0.21 mmol) and methanol (2 ml) were heated at 650C for 60 minutes. The reaction mixture was concentrated in vacuo then azeotroped with toluene (3 x 10 ml) to give a solid residue. The solid residue was dissolved in anhydrous THF (5 ml) and cyclopropylmethylhydroxylamine hydrochloride (49 mg, 0.40 mmol), EDCI (48 mg, 0.25 mmol), HOBt (36 mg, 0.27 mmol) and DIPEA (140 μl, 0.80 mmol) were added. After stirring overnight at 4O0C, the residue was absorbed onto HM- N and purified by flash chromatography (Si-SPE, diethyl ether: MeOH, gradient 98:2 to 95:5) to afford the title compound as an off-white solid (31 mg, 33%). IH NMR (CDCl3, 400MHz) 8.84 (s, IH), 8.62 (s, IH), 8.59 (d, J = 5.9 Hz, IH), 7.99 (s, IH), 7.51 (dd, J = 9.7 Hz, 1.5Hz, IH), 7.43 (d, J = 8.6 Hz, IH), 7.38 (d, J = 5.9 Hz, IH), 6.98 (t, J = 8.4 Hz, IH), 3.90 (d, J = 7.5 Hz, 2H), 1.20 (m, IH), 0.66 (m, 2H), 0.37 (m, 2H).
[00632] EXAMPLE 9: 3-(2-Fluoro-4-iodo-phenylamino)-furor2.3-c1pyridine-2- carboxylic acid ((R)-2,3-dihydroxy-propoxy)-amide
Figure imgf000142_0001
[00633] Step 1 : 3-(2-Fluoro-4-iodo-phenylamino)-furor2,3-c1pyridine-2-carboxylic acid ((R)-2 ,2-dimethyl- [1,31 dioxolan-4-ylmethoxy)-amide
Figure imgf000142_0002
[00634] A mixture of ethyl 3-(2-fluoro-4-iodo-phenylamino)-furo[2,3-c]pyridine-2- carboxylate (220 mg, 0.52 mmol), IN aqueous sodium hydroxide (2.0 ml) and methanol (2.0 ml) was heated at reflux for 15 minutes. The reaction mixture was condensed and azeotroped with toluene (3 x 10 ml) to give a solid residue. The solid residue was dissolved in anhydrous THF (8 ml) and O-((R)-2,2-dimethyl-[l,3]dioxolan-4-ylmethyl)hydroxylamine (149 mg, 1.04 mmol), EDCI (123 mg, 0.64 mmol), HOBt (98 mg, 0.73 mmol) and DIPEA (274 μl, 1.54 mmol) were added. After stirring for 16 hours at ambient temperature, the residue was absorbed onto HM-N and purified by flash chromatography (Si-SPE, diethyl ether: MeOH, gradient 100:0 to 90: 10) to afford the title compound as a brown oil (124 mg, 45%). LCMS (method B): Rx = 3.39 min, M+H+ = 528.
[00635] Step 2; 3-(2-Fluoro-4-iodo-phenylamino)-furor2,3-c1pyridine-2-carboxylic acid ((R)-2,3-dihvdroxy-propoxy)-amide
[00636] 3-(2-Fluoro-4-iodo-phenylamino)-furo[2,3-c]pyridine-2-carboxylic acid ((R)- 2,2-dimethyl-[l,3]dioxolan-4-ylmethoxy)-amide (25 mg, 0.05 mmol) was dissolved in methanol (0.5 ml) and subjected to chromatography (Isolute® SCX-2, MeOH: 2M NH3 in MeOH, gradient 100:0 to 50:50). The residue was then absorbed onto HM-N and purified by flash chromatography (Si-SPE, DCM: MeOH, gradient 95:5 to 80:20) to afford the title compound as a white solid (19 mg, 78%). LCMS (method A): Rx = 6.99 min, M+H+ = 488. IH NMR (d4-Me0H, 400MHz) 8.88 (s, IH), 8.32 (d, J = 5.5 Hz, IH), 7.55 (dd, J = 10.3 Hz, 2.0 Hz, IH), 7.45 (ddd, J = 8.5 Hz, 2.0 Hz, 1.0 Hz, IH), 7.32 (dd, J = 5.5 Hz, 1.0 Hz, IH), 6.89 (t, J = 8.5 Hz, IH), 4.11 (dd, J = 9.9 Hz, 3.5 Hz, IH), 3.97 (m, 2H), 3.63 (m, 2H). [00637] EXAMPLE 10; 3-(2-Fluoro-4-iodo-phenylamino)-furor3.2-c|pyridine-2- carboxylic acid (2-hydroxy-ethoxy)-amide
Figure imgf000143_0001
[00638] Step 1 : 3-(2-Fluoro-4-iodo-phenylamino)-furor3,2-c1pyridine-2-carboxylic acid (2-vinyloxy-ethoxy)-amide
Figure imgf000143_0002
[00639] To a solution of Ethyl 3-(2-fluoro-4-iodo-phenylamino)-furo[3,2-c]pyridine-2- carboxylate (6.50 g, 15.2 mmol) in THF (92 mL) and methanol (31 mL) was added a 1.0 M aqueous solution of sodium hydroxide (31 mL, 31 mmol). The reaction mixture was heated to 65 0C for 1.5 h and then cooled to room temperature and concentrated in vacuo. The resulting residue was azeotroped with toluene (3 x 75 mL), and then suspended in THF (75 mL). O-(2-Vinyloxy-ethyl)-hydroxylamine (1.86 g, 18.0 mmol), N-N-diisopropylethylamine (10.4 mL, 60.0 mmol), EDCI (5.75 g, 30.0 mmol), and HOBt (4.46 g, 33.0 mmol) were then added sequentially, and the reaction mixture stirred 18 hours at room temperature. 18.9 g silica gel was then added and the mixture was concentrated in vacuo. The residue was purified by silica gel chromatography (0-7% Methanol :CH2Cl2) to afford the title compound as a pale yellow solid: 4.40 g, 60%. LCMS (method C): Rx = 2.11 min, M+H+ = 484. IH ΝMR (CDCl3, 400MHz) 8.97 (s, IH), 8.63 (s, IH), 8.60 (d, J = 5.6 Hz, IH), 7.98 (s, IH), 7.52 (dd, J = 9.6, 2.0 Hz, IH), 7.44 (m, IH), 7.39 (dd, J = 6.0, 1.2 Hz, IH), 7.00 (t, J = 8.8 Hz, IH), 6.56 (dd, J = 14.4, 6.8 Hz, 1 H), 4.34 (m, 2H), 4.28 (dd, J = 14.0, 2.0 Hz, 1 H), 4.12
(dd, J = 6.4, 2.0 Hz, 1 H), 4.03 (m, 2 H). [00640] 3-(2-Fluoro-4-iodo-phenylamino)-furor3,2-c1pyridine-2-carboxylic acid (2- hydroxy-ethoxy)-amide
[00641] To a suspension of 3-(2-Fluoro-4-iodo-phenylamino)-furo[3,2-c]pyridine-2- carboxylic acid (2-vinyloxy-ethoxy)-amide (4.40 g, 9.10 mmol) in a mixture of methanol (14.3 mL) and ethanol (51.9 mL) was added a 1.0 M aqueous solution of hydrochloric acid (18.2 mL, 18.2 mmol) at 0 0C. After addition was complete, the reaction mixture was brought to room temperature and stirred for 1.5 h. Solid sodium bicarbonate (4.75 g, 56.5 mmol) was then added portionwise, and stirring was continued for 15 minutes. Silica gel (14 g) was added and the mixture concentrated in vacuo. The residual solid was purified by silica gel chromatography (0-10% methanol:CH2Cl2) to afford the title compound as a pale yellow solid: 4.12 g, 91%. LCMS (method C): Rx = 1.61 min, M+H+ = 458. IH NMR (CDCl3, 400MHz) 8.84 (s, IH), 8.61 (s, IH), 8.60 (s, IH), 7.94 (s, IH), 7.53 (dd, J = 9.6, 2.0 Hz, IH), 7.43 (m, IH), 7.38 (dd, J = 6.0, 1.2 Hz, IH), 7.00 (t, J = 8.4 Hz, IH), 4.30 (b, IH), 4.11 (m, 2 H), 3.83 (b, 2 H).
[00642] EXAMPLE 11: 3-(2-Fluoro-4-iodo-phenylamino)-furor3.2-c1pyridine-2- carboxylic acid (2-vinyloxy-ethoxy)-amide
Figure imgf000144_0001
[00643] EXAMPLE 12: 3-(4-Iodo-phenylamino)-furor3.2-c1pyridine-2-carboxylic acid ((R)-2,3-dihydroxy-propoxy)-amide
Figure imgf000144_0002
[00644] Step 1 : 3-(4-Iodo-phenylamino)-furor3,2-c1pyridine-2-carboxylic acid ((R)-
2,2-dimethyl-rL31dioxolan-4-ylmethoxy)-amide
Figure imgf000144_0003
[00645] A mixture of ethyl 3-(4-iodo-phenylamino)-furo[3,2-c]pyridine-2-carboxylate
(100 mg, 0.24 mmol), IN aqueous sodium hydroxide (260 μl) and ethanol (4 ml) was heated at 65°C for 3 hours. The reaction mixture was condensed and azeotroped with toluene (3 x 20 ml) to give a solid residue. The solid residue was dissolved in anhydrous THF (7 ml), to which EDCI (57 mg, 0.30 mmol) and HOBt (45 mg, 0.33 mmol) were added and the mixture stirred for 30 minutes before O-((R)-2,2-dimethyl)-[l,3]dioxolan-4-ylmethyl)hydroxylamine (71 mg, 0.48 mmol) and DIPEA (125 μl, 0.72 mmol) were finally added. After stirring for 16 hours at ambient temperature, the residue was absorbed onto HM-N and purified by flash chromatography (Si-SPE, cyclohexane: ethyl acetate, gradient 50:50 to 0: 100) to afford the title compound as an off white solid (103 mg, 84%). LCMS (method B): Rx = 2.86 min, M+H+ = 510.
[00646] Step 2: 3-(4-Iodo-phenylamino)-furor3,2-c1pyridine-2-carboxylic acid ((R)-
2.3 -dihydroxy-propoxy)-amide
[00647] 3-(4-Iodo-phenylamino)-furo[3,2-c]pyridine-2-carboxylic acid ((R)-2,2- dimethyl-[l,3]dioxolan-4-ylmethoxy)-amide (100 mg, 0.19 mmol) was dissolved in methanol and subjected to chromatography (Isolute® SCX-2, EtOAc then EtOAc: MeOH: Et3N, 89: 10: 1). The resultant residue was absorbed onto HM-N and purified by flash chromatography (Si-SPE, dichloromethane: MeOH, gradient 100:0 to 90: 10) to afford the title compound as a pale yellow solid (38 mg, 42%). LCMS (method A): Rτ = 6.16 min, M+H+ = 470. IH NMR (d4-Me0H, 400MHz) 8.52 (d, J = 5.9 Hz, IH), 8.48 (s, IH), 7.67 (d, J = 8.8 Hz, 2H), 7.60 (dd, J = 6.0Hz, 0.8 Hz, IH), 7.00 (d, J = 8.7 Hz, 2H), 4.09 (dd, J = 9.9 Hz, 3.4 Hz, IH), 3.93 - 4.00 (m, 2H), 3.61 - 3.64 (m, 2H).
[00648] EXAMPLE 13: 3-(2-Chloro-4-iodo-phenylamino)-furor3.2-c1pyridine-2- carboxylic acid ((R)-2.3-dihydroxy-propoxy)-amide
Figure imgf000145_0001
[00649] Step 1 : 3-(2-Chloro-4-iodo-phenylamino)-furor3,2-clpyridine-2-carboxyric acid ((R)-2 ,2-dimethyl- [1,31 dioxolan-4-ylmethoxy)-amide
Figure imgf000145_0002
[00650] A mixture of ethyl 3-(2-chloro-4-iodo-phenylamino)-furo[3,2-c]pyridine-2- carboxylate (115 mg, 0.26 mmol), IN aqueous sodium hydroxide solution (0.27 ml, 0.27 mmol) and industrialized methylated spirits (3.0 ml) were heated at 650C for 60 minutes. The reaction mixture was concentrated then azeotroped with toluene (3 x 10 ml) to give a solid residue. The resultant solid residue was dissolved in anhydrous THF (5 ml), to which 0-((R)- 2,2-dimethyl-[l,3]dioxolan-4-ylmethyl)hydroxylamine (75 mg, 0.51 mmol), EDCI (65 mg, 0.34 mmol), HOBt (49 mg, 0.36 mmol) and DIPEA (175 μl, 1.02 mmol) were added. The reaction mixture was stirred for 48h, and was then concentrated under reduced pressure. The resultant residue was absorbed onto HM-N and purified by flash chromatography (Si-SPE, dichloromethane: methanol, gradient 100:0 to 95:5) to afford the title compound as a yellow oil (119 mg, 84%). LCMS (method B): Rx = 3.14 min, M+H+ = 544. [00651] Step 2: 3-(2-Chloro-4-iodo-phenylamino)-furor3,2-clpyridine-2-carboxyric acid ((R)-2,3-dihydroxy-propoxy)-amide
[00652] 3-(2-Chloro-4-iodo-phenylamino)-furo[3,2-c]pyridine-2-carboxylic acid ((R)-
2,2-dimethyl-[l,3]dioxolan-4-ylmethoxy)-amide (119 mg, 0.22 mmol) was dissolved in methanol (5.0 ml) and loaded onto an Isolute® SCX-2 cartridge (5g). The cartridge was then washed with methanol (15 ml) and the desired product was subsequently eluted using 2M NH3 in MeOH. The eluant was collected and concentrated to give a residue. The residue was absorbed onto HM-N and purified by flash chromatography (Si-SPE, dichloromethane: methanol, gradient 100:0 to 90: 10) to afford the title compound as a white solid (20 mg, 18%). LCMS (method A): Rx = 7.02 min, M+H+ = 504. IH NMR (d4-MeOH, 400MHz) 8.52 (d, J= 6.2 Hz, IH), 8.52 (d, J = 0.9 Hz, IH), 7.81 (d, J = 2.0 Hz, IH), 7.61 (dd, J = 6.2 Hz, 0.9 Hz, IH), 7.58 (dd, J = 8.5 Hz, 2.0 Hz, IH), 7.01 (d, J = 8.5 Hz, IH), 4.09-4.05 (m, IH), 3.98- 3.88 (m, 2H), 3.60-3.58 (m, 2H).
[00653] EXAMPLE 14: 3-(2.6-Difluoro-4-iodo-phenylamino)-furor3.2-c1pyridine-2- carboxylic acid ((R)-2.3-dihydroxy-propoxy)-amide
Figure imgf000146_0001
[00654] Step 1 : 3-(2.6-Difluoro-4-iodo-phenylamino)-furor3.2-clpyridine-2-carboxylic acid ((R)-2 ,2-dimethyl- [1,31 dioxolan-4-ylmethoxy)-amide
Figure imgf000146_0002
[00655] A mixture of ethyl 3-(2,6-difluoro-4-iodo-phenylamino)-furo[3,2-c]pyridine-
2-carboxylate (137 mg, 0.31 mmol), IN aqueous sodium hydroxide solution (0.32 ml, 0.32 mmol) and industrialized methylated spirits (5.0 ml) were heated at 650C for 60 minutes. The reaction mixture was concentrated then azeotroped with toluene (2 x 10 ml) to give a solid residue. The solid residue was dissolved in anhydrous THF (5 ml) before O-((R)-2,2- dimethyl-[l,3]dioxolan-4-ylmethyl)hydroxylamine (89 mg, 0.61 mmol), EDCI (77 mg, 0.40 mmol), HOBt (58 mg, 0.43 mmol) and DIPEA (213 μl, 1.22 mmol) were added. The reaction mixture was stirred for 16 hours and concentrated under reduced pressure. The residue was absorbed onto HM-N and purified by flash chromatography (Si-SPE, dichloromethane: methanol, gradient 100:0 to 95:5) to afford the title compound as a yellow oil (63 mg, 37%). LCMS (method B): Rx = 2.87 min, M+H+ = 546.
[00656] Step 2: 3-(2,6-Difluoro-4-iodo-phenylamino)-furor3,2-c1pyridine-2-carboxylic acid ((R)-2.3-dihydroxy-propoxy)-amide
[00657] 3-(2,6-Difluoro-4-iodo-phenylamino)-furo[3,2-c]pyridine-2-carboxylic acid
((R)-2,2-dimethyl-[l,3]dioxolan-4-ylmethoxy)-amide (63 mg, 0.11 mmol) was dissolved in methanol (4.0 ml) and loaded onto an Isolute® SCX-2 cartridge (5g). The cartridge was then washed with methanol (15 ml) and the desired product was subsequently eluted using 2M NH3 in MeOH. The eluant was collected and concentrated to give a residue. The resultant residue was absorbed onto HM-N and purified by flash chromatography (Si-SPE, dichloromethane: methanol, gradient 100:0 to 90: 10) to afford the title compound as a white solid (17 mg, 31%). LCMS (method A): Rx = 5.97 min, M+H+ = 506. IH NMR (d4-MeOH, 400MHz) 8.48 (d, J= 6.0 Hz, IH), 8.26 (s, IH), 7.55 (d, J = 6.0 Hz, IH), 7.54-7.49 (m, 2H), 4.08-4.05 (m, IH), 3.96-3.87 (m, 2H), 3.60-3.58 (m, 2H).
[00658] EXAMPLE 15: 3-(2.5-Difluoro-4-iodo-phenylamino)-furor3.2-c1pyridine-2- carboxylic acid ((R)-2.3-dihydroxy-propoxy)-amide
Figure imgf000147_0001
[00659] Step 1 : 3-(2,5-Difluoro-4-iodo-phenylamino)-furor3,2-c1pyridine-2-carboxylic acid ((R)-2 ,2-dimethyl- [1,31 dioxolan-4-ylmethoxy)-amide
Figure imgf000147_0002
[00660] A mixture of ethyl 3-(2,5-difluoro-4-iodo-phenylamino)-furo[3,2-c]pyridine-
2-carboxylate (163 mg, 0.37 mmol), IN aqueous sodium hydroxide solution (0.38 ml, 0.38 mmol) and industrialized methylated spirits (4.0 ml) were heated at 650C for 30 minutes. The reaction mixture was concentrated in vacuo then azeotroped with toluene (2 x 10 ml) to give a solid residue. The solid residue was dissolved in anhydrous THF (5 ml) before O-((R)-2,2- dimethyl-[l,3]dioxolan-4-ylmethyl)hydroxylamine (106 mg, 0.72 mmol), EDCI (91 mg, 0.470 mmol), HOBt (69 mg, 0.51 mmol) and DIPEA (250 μl, 1.45 mmol) were added. The reaction mixture was stirred for 16 hours and concentrated under reduced pressure. The residue was absorbed onto HM-N and purified by flash chromatography (Si-SPE, dichloromethane: methanol, gradient 100:0 to 95:5) to afford the title compound as a yellow oil (152 mg, 76%). LCMS (method B): Rx = 3.01 min, M+H+ = 546.
[00661] Step 2: 3-(2,5-Difluoro-4-iodo-phenylamino)-furor3,2-c1pyridine-2-carboxylic acid ((R)-2.3-dihydroxy-propoxy)-amide
[00662] 3-(2,5-Difluoro-4-iodo-phenylamino)-furo[3,2-c]pyridine-2-carboxylic acid
((R)-2,2-dimethyl-[l,3]dioxolan-4-ylmethoxy)-amide (145 mg, 0.27 mmol) was dissolved in methanol (5.0 ml) and loaded onto an Isolute® SCX-2 cartridge (5g). The cartridge was then washed with methanol (15 ml) and the desired product was subsequently eluted using 2M NH3 in MeOH. The eluant was collected and concentrated to give a residue. The residue was absorbed onto HM-N and purified by flash chromatography (Si-SPE, dichloromethane: methanol, gradient 100:0 to 90: 10) to afford the title compound as a white solid (75 mg, 56%). LCMS (method A): Rx = 6.49 min, M+H+ = 506. IH NMR (d4-MeOH, 400MHz) 8.61 (d, J= 0.9 Hz, IH), 8.52 (d, J=6.1 Hz, IH), 7.61 (dd, J = 6.1 Hz, 0.9 Hz, IH), 7.59 (dd, J=IO. IHz, 5.7 Hz, 2H), 6.94 (dd, J=8.4 Hz, 7.5 Hz, IH), 4.07-4.03 (m, IH), 3.96-3.87 (m, 2H), 3.63-3.55 (m, 2H).
[00663] EXAMPLE 16: 4-{r3-(2-Fluoro-4-iodo-phenylamino)-furor3.2-c1pyridine-2- carbonyliaminooxyl-piperidine-l-carboxylic acid tert-butyl ester
Figure imgf000148_0001
[00664] A mixture of 3-(2-fluoro-4-iodo-phenylamino)-furo[3,2-c]pyridine-2- carboxylic acid (398 mg, 1 mmol), HOBT (190 mg, 1.4 mmol) and EDCI (240 mg, 1.25 mmol) in THF (5 ml) was stirred for Ih. To this mixture was added DIPEA (530 μl, 3.0 mmol) and 4-aminooxy-piperidine-l-carboxylic acid tert-butyl ester (432 mg, 2.0 mmol). After stirring overnight, the reaction mixture was concentrated in vacuo, diluted with ethyl acetate (30 ml) and washed with saturated NaHCO3 solution (10 ml). The organic layer was isolated, dried over sodium sulfate and concentrated in vacuo. Purification of the resultant residue by flash chromatography (Si-SPE, dichloromethane methanol, gradient 100% to 95 5) afforded the title compound as a pale yellow foam (285 mg, 47%) LCMS (method A) RT = 10 43 mm, M+H+ = 597 IH NMR (CDCl3, 400MHz) 11 86 (s, IH), 8 15 (s, IH), 7 66 (dd, J = 10 8 Hz, 2 0 Hz, IH), 7 61 (d, J = 8 6 Hz, IH), 7 51 (m, IH), 7 43 (dd, J = 8 8 Hz, IH), 7 32 (d, J = 7 7 Hz, IH), 7 27 (m, IH), 6 86 (t, J = 8 8 Hz, IH), 4 88 (d (br), J = 3 4 Hz, IH), 4 60 (t (br), J = 5 5 Hz, IH), 3 17 (ddd, J= 13 6, 8 8 3 7 Hz, 2H), 1 97-2 03 (m, 2H), 1 7- 1 81 (m, 2H), 1 63 (s, 9H)
[00665] EXAMPLE 17; 3-(2-Fruoro-4-iodo-phenylamino)-furor3.2-clpyridine-2- carboxyhc acid (2-morpholin-4-yl-ethoxy)-amide
Figure imgf000149_0001
[00666] A mixture of 3-(2-fluoro-4-iodo-phenylamino)-furo[3,2-c]pyπdine-2- carboxyhc acid (199 mg, 0 5 mmol), HOBt (95 mg, 0 7 mmol) and EDCI (125 mg, 0 65 mmol) in THF (2 ml) was stirred for Ih To this mixture was added DIPEA (270 μl, 1 5 mmol) and O-(2-(tetrahydropyran-4-yl)ethyl)hydroxylamine (146 mg, 1 0 mmol) After stirring overnight, the solution was concentrated in vacuo, diluted with ethyl acetate (30 ml) and washed with saturated NaHCO3 solution (10 ml) The organic layer was isolated, dried over sodium sulfate and concentrated in vacuo Purification of the resultant residue by flash chromatography (Si-SPE, dichloromethane methanol, gradient 100 0 to 95 5) afforded the title compound as a foam which crystallised from ether/dichloromethane (75 mg, 28%) LCMS (method A) Rx = 5 67 mm, M+H+ = 527 IH NMR (CDCl3, 400MHz) 8 77 (s, IH), 8 63 (d, J = 0 7 Hz, IH), 8 59 (d, J = 5 8Hz, IH), 7 99 ( s, IH), 7 51 (dd, J = 9 7 Hz, 2 0 Hz, IH), 7 42-7 46 (m, IH), 7 37 (dd, J = 6 0, 1 2 Hz, IH), 6 99 (t, J = 8 5 Hz, IH), 4 20-4 12 (m, 2H), 3 92-3 80 (m, 2H), 3 17 (ddd, J = 13 6, 8 8, 3 7 Hz, 2H), 2 03-1 97 (m, 2H), 1 81-1 7 (m, 2H), 1 68-1 61 (m, 2H)
[00667] EXAMPLE 18; 7-Bromo-3-(2-fluoro-4-iodo-phenylamino)-furor3.2- cipyridine-2-carboxylic acid ((R)-2.3-dihydroxy-propoxy)amide
Figure imgf000149_0002
[00668] Step 1 7-Bromo-3-(2-fluoro-4-iodo-phenylamino)-furo|"3.2-c]pyridrne-2- carboxylic acid ((R)-2,2-dimethyl-r 1 ,3"|dioxolan-4-yrmethoxy)-amide
Figure imgf000150_0001
[00669] A mixture of 7-bromo-3-(2-fluoro-4-iodo-phenylamino)-furo[3,2-c]pyridine-
2-carboxylic acid (50 mg, 0.21 mmol) and carbonyl diimidazole (35 mg, 0.21 mmol) in acetonitrile (2 ml) was heated at 5O0C for 4 hours. The reaction was then treated with a solution of O-((R)-2,2-dimethyl-[l,3]dioxolan-4-ylmethyl)-hydroxylamine (46 mg, 0.36 mmol) in acetonitrile (1 ml) and heated at 8O0C for 3.5 hours before cooling and allowing to stand at room temperature. The reaction mixture was filtered, washed with ethyl acetate before the filtrate was collected and concentrated in vacuo. Purification of the resultant residue by flash chromatography (Si-SPE, dichloromethane: ethyl acetate, gradient 1:0 to 4: 1 to 0: 1, then methanol) afforded the title compound as a light brown solid (11 mg, 20%). LCMS (method B): Rx = 3.55 min, M+H+ = 606/608.
[00670] Step 2: 7-Bromo-3-(2-fluoro-4-iodo-phenylamino)-furor3,2-clpyridine-2- carboxylic acid ((R)-2,3-dihydroxy-propoxy)amide
[00671] 7-Bromo-3-(2-fluoro-4-iodo-phenylamino)-furo[3,2-c]pyridine-2 -carboxylic acid ((R)-2,2-dimethyl-[l,3]dioxolan-4-ylmethoxy)-amide (40mg, 0.066mmol) was dissolved in 0.067M methanolic HCl (2.79ml, 0.198mmol) and stirred at room temperature for 40 mins. The reaction mixture was concentrated in vacuo then azeotroped with toluene (2 x 15ml). The resultant residue was dissolved in IMS (4 ml), and then potassium carbonate was added, before stirring at room temperature for 4 mins. The reaction mixture was filtered and washed with IMS, before the filtrate was evaporated in vacuo to afford a solid. The resultant solid was triturated with acetonitrile to afford the desired product as a cream solid (29 mg, 77%). LCMS (Method A): Rx = 9.06 min, M+H+ = 566/568. 1H NMR (CD3OD): 8.68 (IH, s, br), 8.42 (IH, s, br), 7.61 (IH, dd, J = 10.2, 1.9Hz), 7.52 (IH, m), 7.07 (IH, t, J = 8.6Hz), 4.12 (IH, dd, J = 10.0, 3.4Hz), 3.99 (IH, dd, J = 10.0, 6.8Hz), 3.96 (IH, m), 3.63 (2H, m). [00672] EXAMPLE 19: 5-(2-Fluoro-4-iodo-phenylamino)-furor2.3-d1pyrimidine-6- carboxylic acid ((R)-2,3-dihvdroxy-propoxy)-amide
Figure imgf000150_0002
[00673] Step 1 : 5-(2-fluoro-4-iodo-phenylamino)-furo[2.3-dlpyrimidine-6-carboxylic acid ((R)-2 ,2-dimethyl- [1,31 dioxolan-4-ylmethoxy)-amide
Figure imgf000151_0001
[00674] A mixture of ethyl 5-(2-fluoro-4-iodo-phenylamino)-furo[2,3-d]pyrimidine-6- carboxylate (300 mg, 0.70 mmol), IN aqueous sodium hydroxide solution (0.75 ml, 0.75 mmol) and industrialized methylated spirits (8.0 ml) were heated at 650C for 30 minutes. The reaction mixture was concentrated then azeotroped with toluene (3 x 10 ml) to give a solid residue. The solid residue was dissolved in anhydrous THF (5 ml) and O-((R)-2,2-dimethyl- [l,3]dioxolan-4-ylmethyl)hydroxylamine (106 mg, 0.72 mmol), EDCI (91 mg, 0.47 mmol), HOBt (69 mg, 0.51 mmol) and DIPEA (250 μl, 1.45 mmol) were added. The reaction mixture was stirred for 18h before being concentrated under reduced pressure. The resultant residue was absorbed onto HM-N and purified by flash chromatography (Si-SPE, dichloromethane: methanol, gradient 100:0 to 95:5) to afford the title compound (124 mg, 67%). LCMS (method B): Rx = 3.46 min, M+H+ = 529.
[00675] Step 2: 5-(2-Fluoro-4-iodo-phenylamino)-furor2,3-d1pyrimidine-6-carboxylic acid ((R)-2.3-dihydroxy-propoxy)-amide
[00676] To a suspension of 5-(2-fluoro-4-iodo-phenylamino)-furo[2,3-d]pyrimidine-6- carboxylic acid ((R)-2,2-dimethyl-[l,3]dioxolan-4-ylmethoxy)-amide (124 mg, 0.23 mmol) in methanol (5.0 ml) was added concentrated hydrochloric acid (10 drops) and the mixture was stirred for 1 hour. The reaction mixture was then concentrated under reduced pressure, dissolved in methanol (5ml), and potassium sodium carbonate (-200 mg) was added. This mixture was stirred for 5 minutes, absorbed on HM-N and purified by flash chromatography (Si-SPE, dichloromethane: methanol, gradient 100:0 to 0: 100) to afford the title compound as a white solid (60 mg, 54%). LCMS (method A): Rx = 7.85 min, M+H+ = 489. IH NMR (d6- DMSO, 400MHz) 8.96 (s, IH), 8.77 (s, IH), 7.64 (dd, J =10.7 Hz, 1.9 Hz, IH), 7.43 (ddd, J = 8.5 Hz, 1.9 Hz, 0.9 Hz, IH), 6.91 (dd, J = 8.5 Hz, 8.5 Hz, IH), 3.86-3.71 (m, 3H), 3.40-3.30 (m, 4H).
[00677] EXAMPLE 97: 3-(2-Fluoro-4-iodophenylamino)furor3.2-c1pyridine-2- carboxylic acid (l-methylpiperidin-4-yloxy) amide
Figure imgf000152_0001
[00678] A mixture of 3-(2-fmoro-4-iodo-phenylamino)-furo[3,2-c]pyridine-2- carboxylic acid(piperidin-4-yloxy)-amide (80 mg, 0.16 mmol), pyridinium p-toluenesulfonic acid (40 mg, 0.16 mmol) and formaldehyde (0.05 ml, 1OM aq. soln., 0.48 mmol) was suspended in methanol (0.5 ml) and was stirred under argon for 16 hours. Sodium cyanoborohydride (30 mg, 0.47 mmol) was added and the resulting solution was stirred for Ih. The solvent was evaporated and the resultant residue partitioned between ethyl acetate and sodium bicarbonate, the organic layer was isolated, washed with brine, dried (Na2SO4) and evaporated. The resultant product was purified by flash chromatography (Si-SPE 2M methanolic ammonia: DCM gradient 0: 100 to 10: 100) to afford the product as a pale yellow solid (50 mg, 61% yield). LCMS (method A): Rx = 5.49min; M+H+ 511; IH NMR (CDCl3) 1.88 (2H, m), 2.08 (2H, m), 2.22 (2H, m), 2.31 (3 H, s), 2.78 (2H, m), 4.08 (IH, m), 6.98 (IH, t, J = 8.5Hz), 7.37 (IH, dd, J = 5.9, 1.0Hz), 7.44 (IH, m), 7.51 (IH, dd, J = 9.8, 1.9Hz), 7.99 (IH, s), 8.59 (IH, d, J = 5.9Hz), 8.63 (IH, d, J = 1.0Hz).
[00679] EXAMPLE 98: 3-(2-Fluoro-4-iodo-phenylamino)furor3.2-c1pyridine-2- carboxylic acid (2-dimethylaminoethoxy) amide
Figure imgf000152_0002
[00680] A mixture of 3-(2-fluoro-4-iodo-phenylamino)-fur o[3,2-c]pyridine-2-carboxylic acid (2-methylamino-ethoxy)-amide (45 mg, 99 mmol), formaldehyde (8 μL, 107 mmol), pyridinium p-toluenesulfonic acid (25 mg, 97 mmol) in ethanol (1 mL) was stirred at 0-50C for 30min. Sodium cyanoborohydride (7 mg, 106 mmol) was added and the solution was stirred for 30 min at room temperature. HCl (IM, 200 μL) was added and the solution was loaded onto an Isolute® SCX-2 cartridge (2 g) eluting with methanolic ammonia. The appropriate fractions were combined and concentrated to provide a residue which was further purified by flash chromatography (Si-SPE gradient 2M methanolic ammonia: DCM 0: 100 to 10: 100) to afford the product as a white foam. (23 mg). LCMS (method A): Rx = 5.12min M+H+ 484; IH NMR (CD3OD) 2.70 (6H, s), 3.11 (2H, m), 4.15 (2H, m), 6.80 (IH, t, J = 8.7Hz), 7.42 (IH, m), 7.53 (IH, dd, J = 10.6, 1.9Hz), 7.61 (IH, dd, J
= 5.9, 0.9Hz), 8.47 (IH, d, J = 5.9Hz), 8.56 (IH, d, J = 0.9Hz).
[00681] EXAMPLE 99: (2-Fluoro-4-iodo-phenylamino)-N-tert-butoxyfuror3.2- clpyridine-2-carboxamide
Figure imgf000153_0001
A mixture of 3-(2-fluoro-4-iodo-phenylamino)-furo[3,2-c]pyridine-2-carboxylic acid ethyl ester (251 mg, 0.59 mmol), IN aqueous NaOH solution (1.77 ml, 1.77 mmol), methanol (15 ml) and tetrahydrofuran (15 ml) was refluxed for 2 hours. The reaction mixture was concentrated in vacuo then the resultant residue was azeotroped with toluene (3 x 15 ml) to give a solid residue. The solid residue was triturated with ether (3 x 10 ml), and the ether layers were discarded. The resultant solid residue was dried under vacuum. The solid residue was then dissolved in anhydrous DMF (5 ml) before 0-tertbutylhydroxylamine hydrochloride (58 mg, 0.56 mmol), HATU (270 mg, 0.71 mmol) and DIPEA (470 μl, 2.36 mmol) were added. After stirring for 18 hours at ambient temperature the solvent was evaporated and the resultant residue was purified by preparative HPLC to afford the title compound as a white solid (80 mg, 23%). LCMS (method E): Rx = 2.30 min, M+H+ = 470. 1H NMR (CDCl3, 400MHz) 8.82 (d, IH), 8.69 (s, IH), 8.39 (s, IH), 8.15 (s, IH), 7.73 (d, IH), 7.59 (dd, IH), 7.51 (d,m, IH), 6.85 (t, IH), 1.4 (2, IH).
[00682] EXAMPLE 100; G-(2-fluoro-4-iodophenylamino)furor3.2-c1pyridin-2 -ylYl- oxothiazolidin-3 -vDmethanone
Figure imgf000153_0002
[00683] (3-(2-Fluoro-4-iodophenylamino)furo[3,2-c]pyridin-2-yl)(thiazolidin-3- yl)methanone was dissolved in methanol (1 ml) and THF (1 ml) and cooled to -5°C. Oxone (21 mg, 0.035 mmol) in water (0.5 ml) was added, and the reaction was stirred and warmed to room temperature over 1 hour. The reaction was diluted with ethyl acetate (3 ml) and water (1 ml) and solids decanted. The layers were separated, the aqueous layer extracted with ethyl acetate, and the combined organic layers were washed with sat. aqueous NaHCO3, and brine, dried (MgSO4), and concentrated. . The resulting oil was purified by reverse phase HPLC to provide the TFA salt of the title compound as a yellow solid (6.8 mg). LCMS (method D): Rx = 2.45 min M+H+ = 486.
[00684] EXAMPLE 101: G-(2-fluoro-4-iodophenylamino)furor3.2-c1pyridin-2- yl)( 1 , 1 -dioxothiomorpholin-4-yl)methanone
Figure imgf000154_0001
[00685] (3-(2-Fluoro-4-iodophenylamino)furo [3,2-c]pyridin-2-yl)(thiomorpholin-4- yl)methanone (19 mg, 0.039 mmol) was dissolved in methanol (1 ml) and THF (1 ml) and cooled to -5°C. Oxone (30 mg, 0.049 mmol) in water (0.5 ml) was added, and the reaction was stirred while warming to room temperature over 1 hour. The reaction was diluted with ethyl acetate (3 ml) and water (1 ml) and solids decanted. The layers were separated, the aqueous layer extracted with ethyl acetate, and the combined organic layers were washed with saturated aqueous NaHCθ3, and brine, dried (MgSO4), and concentrated. . The resulting oil was purified by reverse phase HPLC to provide the TFA salt of the title compound as a yellow solid (1.5 mg). LCMS (method E) Rx = 4.17, M+H+ = 516. [00686] EXAMPLE 102: (3-(2.5-difluoro-4-(4-pyrazolyl)phenylamino)furor3.2- c1pyridin-2-yl)((R)-3 -hydroxypyrrolidin- 1 -vDmethanone
Figure imgf000154_0002
A degassed solution of (3-(4-bromo-2,5-difluorophenylamino)furo[3,2-c]pyridin-2-yl)((R)-3- hydroxypyrrolidin-l-yl)methanone (53 mg, 0.12 mmol), l-Boc-pyrazole-4-boronic acid pinacol ester (53 mg, 0.18 mmol), Pd(PPh3)4 (7.0 mg, 0.0061 mmol), and Na2CO3 (29 mg, 0.27 mmol) in dimethoxyethane (2.0 ml), ethanol (0.7 ml), and water (0.7 ml) was heated at reflux overnight. The reaction mixture was cooled to ambient temperature, filtered, and the solid was washed with ethyl acetate and dried to provide the crude product as a tan solid (41 mg, 80%). LCMS (method D): Rx = 1.44 min M+H+ = 426). 1H NMR (DMSO-d6, 400MHz) 8.77-8.82 (m, 2H), 8.66-8.68 (m, IH), 8.07 (app s, 2H), 7.91-7.93 (m, 1 H), 7.73-7.78 (m, 1 H), 7.10-7.15 (m, 1 H), 4.32-4.41 (m, 1 H), 3.89-4.04(m, 2 H), 3.76-3.80 (m, 1 H), 3.45-3.62 (m, 2 H), 1.80-2.02 (m, 2 H).
[00687] EXAMPLE 104: 2-Dimethylcarbamoyl-3-(2-fruoro-4-iodo-phenylamino)- furor3.2-c1pyridine-7-carboxylic acid ethyl ester
Figure imgf000155_0001
[00688] To a solution of 2-dimethylcarbamoyl-3-(2-fluoro-4-trimethylsilanyl- phenylamino)furo[3,2-c]pyridine-7-carboxylic acid ethyl ester (84 mg, 0.19 mmol) in dichloromethane (3 ml) at -1O0C was added iodine monochloride (0.38 ml, 0.38 mmol, IM solution in dichloromethane) and the solution was stirred at this temperature for 1 hour. A saturated solution of sodium thiosulfate (5 ml) was added and the mixture was poured into saturated sodium thiosulfate (15 ml). The aqueous layer was isolated then extracted with dichloromethane (2 x 25 ml), before the combined organic layers were washed with brine, dried over magnesium sulfate and concentrated in vacuo. Purification of the resultant residue by flash chromatography (Si-SPE, dichloromethane : t-butyl methyl ether gradient 1 :0 to 1:3) afforded the title compound as a yellow waxy solid (87 mg, 92%). LCMS (method B): Rτ = 3.97 min, M+H+ = 498.
[00689] EXAMPLE 105: 3-(2-Fluoro-4-iodo-phenylamino)-7-hvdroxymethyl- furor3,2-c1pyridine-2-carboxylic acid dimethylamide
Figure imgf000155_0002
[00690] To a solution of 2-dimethylcarbamoyl-3-(2-fluoro-4-iodo-phenylamino)- furo[3,2-c]pyridine-7-carboxylic acid ethyl ester (96 mg, 0.193 mmol) in THF (4 ml) at -4O0C was added dropwise lithium triethylborohydride (0.41 ml, 0.41 mmol, IM solution in THF). The resulting mixture was stirred at -4O0C for 30 minutes before quenching the reaction with the addition of saturated ammonium chloride (20 ml). The aqueous layer was isolated then extracted with dichloromethane (3 x 20 ml) before the combined organic layers were dried over magnesium sulfate and concentrated in vacuo. Purification of the resultant residue by flash chromatography (Si-SPE, dichloromethane : ethyl acetate gradient 1:0 to 0: 1 then ethyl acetate : methanol 85: 15) afforded crude material. The crude material was triturated in methanol to afford the title compound as a white solid (34 mg, 39%). 1H NMR (DMSO-D6, 400 MHz) 3.07 (6H, s, br), 4.83 (2H, d, J = 5.6 Hz), 5.47 (IH, t, J = 5.6 Hz), 6.87 (IH, t, J = 8.7 Hz), 7.43 (IH, m), 7.66 (IH, dd, J = 10.8, 2.0 Hz), 8.54 (2H, m), 8.56 (IH, s). LCMS (method A): Rx = 6.74 min, M+H+ = 456.
[00691] EXAMPLE 106: 3-(2-Fluoro-4-iodo-phenylamino)-7-phenoxymethyl- furor3,2-c1pyridine-2-carboxylic acid dimethylamide
Figure imgf000156_0001
[00692] To a solution of 3-(2-fluoro-4-iodo-phenylamino)-7-hydroxymethyl-furo[3,2- c]pyridine-2-carboxylic acid dimethylamide (34 mg, 0.075 mmol) and triphenylphosphine (20 mg, 0.075 mmol) in THF (3 ml) was added phenol (7.75 mg, 0.083 mmol) and DIAD (18.5 μl, 0.094 mmol) and the mixture was stirred at room temperature for 21 hours. The reaction mixture was diluted with ethyl acetate (40 ml) washed with IM NaOH (15 ml) and brine (15 ml). The organic layer was isolated, dried over magnesium sulfate and concentrated in vacuo. Purification of the resultant residue by flash chromatography (Si-SPE, dichloromethane : ethyl acetate gradient 1 :0 to 0: 1 then ethyl acetate : methanol 85: 15) afforded crude material. Repurifϊcation by flash chromatography (Si-SPE, ethyl acetate : dichloromethane : cyclohexane 1:4: 1) afforded the title compound as a white solid (12 mg, 30%). 1H NMR (CDCl3, 400 MHz) 3.26 (6H, s, br), 5.37 (2H, s), 6.94 (IH, t, J = 8.6 Hz), 7.02 (3H, m), 7.33 (2H, m), 7.41 (IH, m), 7.49 (IH, dd, J = 9.9, 1.9 Hz), 8.55 (IH, s), 8.66 (2H, apparent s). LCMS (method A): Rx = 12.30 min, M+H+ = 532. [00693] EXAMPLE 107: 7-chloro-3-(2-fluoro-4-iodo-phenylamino)-furor3.2- clpyridine-2-carboxylic acid sodium salt
Figure imgf000156_0002
[00694] To a suspension of 7-chloro-3-(2-fluoro-4-iodo-phenylamino)-furo[3,2- c]pyridine-carboxylic acid ethyl ester (100 mg, 0.277 mmol) in methanol (7 ml) was added IM NaOH (0.32 ml, 0.32 mmol) and the mixture was stirred at 550C for 3 hours. The resultant suspension was cooled to room temperature and stirred for 17 hours then heated at 550C for 2.5 hours. The reaction mixture was concentrated in vacuo and the residue azeotroped with toluene (2 x 20 ml). The resultant residue was then triturated in water and filtered to afford the title compound as a white solid (74 mg, 75%). 1H NMR (CD3OD, 400 MHz) 6.95 (IH, t, J = 8.6 Hz), 7.44 (IH, ddd, J = 8.4, 1.8, 1.2 Hz) 7.54 (IH, dd, J = 10.4, 1.9 Hz), 8.48 (IH, s), 8.49 (IH, s). LCMS (method A): Rx = 10.57 min, M+H+ = 433. [00695] EXAMPLE 108; 7-Chloro-3-(2-fluoro-4-iodo-phenylamino)-furor3.2- cipyridine-2-carboxylic acid amide
Figure imgf000157_0001
[00696] To a solution of 7-chloro-3-(2-fluoro-4-iodo-phenylamino)-furo[3,2- c]pyridine-2-carboxylic acid sodium salt (60 mg, 0.110 mmol) and ammonium chloride (17.5 mg, 0.33 mmol) in DMF (1.5 ml) was added HATU (84 mg, 0.22 mmol) and diisopropylethylamine (75 μl, 0.44 mmol) and the resulting solution was stirred at room temperature for 3 hours. The reaction mixture was diluted with ethyl acetate and washed with water then saturated sodium bicarbonate and then brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The resultant residue was triturated in acetonitrile to afford the title compound as a yellow solid (14 mg, 30%). 1H NMR (CDCl3, 400 MHz) 5.69 (IH, s, br), 6.30 (IH, s, br), 6.98 (IH, t, J = 8.4 Hz), 7.46 (IH, m), 7.53 (IH, dd, J = 9.7, 1.9 Hz), 8.03 (IH, s), 8.46 (IH, s), 8.56 (IH, s). LCMS (method A): Rx = 10.46 min, M+H+ = 432.
[00697] EXAMPLE 109: 3-(2-Fluoro-4-iodo-phenylamino)-furor3.2-c1pyridine-2- carboxylic acid (2-methanesulfonylamino-ethoxy)-amide
Figure imgf000157_0002
N-(2-Aminooxy-ethyl)-methanesulfonamide (116 mg, 0.75 mmol), 3-(2-fluoro-4-iodo- phenylamino)-furo[3,2-c]pyridine-2-carboxylic acid (300 mg, 0.75 mmol), EDC (159 mg, 0.83 mmol), HOBT (112 mg, 0.83 mmol) and DIPEA (0.13 ml, 0.75 mmol) were suspended in THF (5 ml) before DMF (5 drops) was added. The reaction was stirred at room temperature for 16 hours. The reaction was concentrated in vacuo and the residue dissolved in ethyl acetate (20 ml) and washed with aqueous saturated sodium bicarbonate solution (20 ml). The aqueous layer was washed with ethyl acetate (2 x 10 ml) and the combined organic extracts were washed with brine and dried over magnesium sulfate and concentrated in vacuo. The resultant residue was subjected to flash chromatography (SiCh, gradient 0-10% methanol in dichloromethane) to yield the title compound as a light yellow solid (120 mg, 49%). 1H NMR (CDCl3, 400 MHz) 8.95 (IH, s), 8.61-8.59 (2H, m), 7.91 (IH, s), 7.54 (IH, dd, J = 9.6, 1.9 Hz), 7.48 (IH, dt, J = 8.4, 1.3 Hz), 7.37 (IH, dd, J = 5.9, 0.8 Hz), 7.03 (IH, t, J = 8.4 Hz), 6.09-6.06 (IH, m), 4.17-4.15 (2H, m), 3.47-3.43 (2H, m), 3.04 (3H, s). LCMS (method A): Rx = 7.34 min, M+H+ = 535.
[00698] EXAMPLE 138: 7-Chloro-3-(2-fluoro-4-iodo-phenylamino)-furor3.2- cipyridine-2- carboxylic acid ((R)-2,3-dihydroxy-propoxy)-amide
Figure imgf000158_0001
[00699] Step 1 : 7-Chloro-3-(2-fluoro-4-iodo-phenylamino)-furor3.2-c1pyridine-2- carboxylic acid
Figure imgf000158_0002
A suspension of 7-chloro-3-(2-fluoro-4-iodo-phenylamino)-furo[3,2-c]pyridine-2-carboxylic acid ethyl ester (150 mg, 0.33 mmol) in IMS (10 ml) was treated with was treated with sodium hydroxide (IM aqueous solution, 0.832 ml) and the reaction mixture heated at 6O0C for 3 hours. The resultant mixture was allowed to cool then concentrated in vacuo, the crude residue treated with water and the mixture adjusted to pH 5 with acetic acid. The resultant suspension was filtered, the residue collected and dried in vacuo to give the title compound as a yellow solid (105 mg, 74%). IH NMR (DMSO-d6 400 MHz) 8.63 (1 H, s), 8.45 (1 H, s), 7.69 (1 H, d, J = 10.28 Hz), 7.49 (1 H, d, J = 8.22 Hz), 7.03 (1 H, s). [00700] Step 2: 7-Chloro-3-(2-fluoro-4-iodo-phenylamino)-furor3.2-c1pyridine-2- carboxylic acid ((R)-2,2-dimethyl-r 1 ,3"|dioxolan-4-yrmethoxy)-amide
Figure imgf000159_0001
[00701] A suspension of 7-chloro-3-(2-fluoro-4-iodo-phenylamino)-furo[3,2- c]pyridine-2-carboxylic acid (177 mg, 0.41 mmol) in dry dichloromethane (6 ml) under an atmosphere of nitrogen was cooled to O0C and treated with DMF (1 drop) and oxalyl chloride (0.102 ml, 1.16 mmol). The reaction mixture was stirred for 1 hour then the solvent removed in vacuo. The resultant residue was re-suspended in dichloromethane and treated dropwise with a solution of O-((R)-2,2-dimethyl-[l,3]dioxolan-4-ylmethyl)-hydroxylamine (101 mg, 0.69 mmol) and DIPEA (0.172 ml, 1.21 mmol) in dichloromethane (4 ml) before being stirred for 3 hours. The reaction mixture was washed (water, brine), dried (Na2SO4), filtered and concentrated in vacuo to give the title compound as a yellow foam (207 mg, 90%). The foam was used in the subsequent step without further analysis or purification. [00702] Step 3: 7-Chloro-3-(2-fluoro-4-iodo-phenylamino)-furor3,2-clpyridine-2- carboxylic acid ((R)-2,3-dihydroxy-propoxy)-amide
[00703] A solution of 7-chloro-3-(2-fluoro-4-iodo-phenylamino)-furo[3,2-c]pyridine-
2-carboxylic acid ((R)-2,2-dimethyl-[l,3]dioxolan-4-ylmethoxy)-amide (280 mg, 0.49 mmol) in methanol/ c.HCl (0.14 ml c.HCl(aq) in 25 ml methanol) was stirred and room temperature until TLC showed no starting material remaining. The reaction mixture was concentrated in vacuo and the residue treated with dichloromethane (11 ml) and triethylamine (0.210 ml), stirring for 20 minutes before re-concentrating in vacuo. The resultant solid residue was subjected to reverse phase HPLC (Phenomenex Luna 5 Cl 8, 0.1% HCO2H in water on a gradient of acetonitrile) to give the title compound as a pale yellow solid (168 mg, 66%). LCMS (method A): Rx = 8.87 min, M+H+ = 522. (CD3OD 400 MHz) 3.61 (IH, dd, J = 11.4, 5.3Hz), 3.65 (IH, dd, J = 11.4, 5.1Hz), 3.94 (IH, m), 3.99 (IH, dd, J = 10.0, 6.8Hz), 4.11 (IH, dd, J = 10.0, 3.5Hz), 7.07 (IH, t, J = 8.6Hz), 7.52 (IH, m), 7.61 (IH, dd, J = 10.2, 1.9Hz), 8.38 (IH, s), 8.56 (IH, s).
[00704] EXAMPLE 139; 7-Fluoro-3-('2-fluoro-4-iodophenylamino)-furor3.2- cipyridine-2-carboxylic acid (2-hydroxyethoxy)amide
Figure imgf000160_0001
[00705] Step 1 : 7-Fluoro-3-(2-fluoro-4-iodophenylamino)-furor3,2-c"|pyridine-2- carboxylic acid
Figure imgf000160_0002
[00706] 7-Fluoro-3-(2-fluoro-4-iodophenylamino)-furo[3,2-c]pyridine-2 carboxylic acid ethyl ester (4.00 g, 9.0 mmol) was stirred as a suspension in ethanol (150 ml) at room temperature before being treated with IM NaOH and heated at 6O0C for 2 hours. The solvent was removed in vacuo and the residue diluted with water (50 ml) and acidified to pH 4 with glacial acetic acid. The resulting solid precipitate was collected by filtration, washed with water and dried at 400C under vacuum (P2O5) to give the title compound (3.75 g quant.). LCMS (method B) Rx 3.51 min M+H+ 417.
[00707] Step 2: 7-Fluoro-3-(2-fluoro-4-iodophenylamino)-furor3,2-clpyridine-2- carboxylic acid (2-vinyloxyethoxy)amide
Figure imgf000160_0003
[00708] 7-Fluoro-3-(2-fluoro-4-iodophenylamino)-furo[3,2-c]pyridine-2 carboxylic acid (3.75 g, 9.0 mmol) was stirred as a suspension in dry DCM (100 ml) under argon at 0 0C, and treated dropwise with oxalyl chloride (2.24 ml, 25.7 mmol), maintaining the temperature below 50C. The resulting mixture was stirred for a further 1 hour before concentration in vacuo. The residue was re-suspended in dry DCM (100 ml) under argon at 0 0C and treated dropwise with a solution O-(2-vinyloxyethyl)hydroxylamine (1.40 g, 14.3 mmol) and diisopropylethylamine (4.70 ml, 3.48g, 27 mmol) in DCM (20 ml). The resulting solution was stirred and allowed to warm to room temperature over 3 hours before being washed with water then saturated saline, dried (MgSO4), filtered and evaporated in vacuo to give a residue which was subjected to flash chromatography (SiC^, gradient 0-10% ethyl acetate in dichloromethane) to give the title compound as a yellow solid (2.41 g 53%). LCMS
(Method B) Rx 3.82 min M+H+ 502.
[00709] Step 3: 7-Fluoro-3-(2-fluoro-4-iodophenylamino)-furor3.2-c1pyridine-2- carboxylic acid (2-hvdroxyethoxy)amide
[00710] 7-Fluoro-3-(2-fluoro-4-iodophenylamino)-furo[3,2-c]pyridine-2 carboxylic acid (2-vinyloxyethoxy)amide (2.41 g, 4.80 mmol) was suspended in ethanol (100 ml) and concentrated hydrochloric acid (2.0 ml) added. The mixture was stirred at room temperature for 1 hour before neutralizing by the addition saturated aqueous sodium hydrogencarbonate solution. The solvent was then removed in vacuo, and the resultant residue was dissolved in
DCM, washed water, dried (MgSO4) filtered and evaporated in vacuo to give a crude residue which was subjected to flash chromatography (Siθ2, gradient 0 to 2% methanol in dichloromethane) followed by recrystallisation (aqueous methanol) to give the title compound as yellow needles (0.837 g, 36%). LCMS (method A): Rx 9.15 min, M+H+ 476;
IH NMR (DMSO-d6, 400 MHz) 3.60 (2 H, m), 3.88-3.93 (2H, m), 4.70 (1 H, br s), 7.02 (IH, t, J = 8.68 Hz), 7.46-7.49 (IH, m), 7.68 (IH, dd, J = 10.55, 1.92 Hz), 8.36 (IH, s), 8.40 (IH, s), 8.64 (IH, d, J = 2.58 Hz), 11.95 (1 H, s).
[00711] EXAMPLES 20-96 and 111-159
[00712] Compounds in Tables 2. 3. and 4 were prepared by general methods outlined below:
[00713] Amides and hydroxamates were prepared from the appropriate acid by using the coupling general method described below. In some cases the intermediate acid was not isolated, the coupling reaction performed on the crude carboxylate salt produced by following the saponification general method.
[00714] Saponification general method
[00715] A mixture of carboxylic acid ester, IN aqueous NaOH (1-2 eq.) and EtOH was heated at 700C for 1 hour. The reaction mixture was concentrated in vacuo and azeotroped with toluene to give the crude carboxylate salt.
[00716] Coupling general method
[00717] The appropriate carboxylic acid or carboxylate salt was suspended in anhydrous THF before the appropriate hydroxylamine or amine (l-4eq.), EDCI (l-2eq.) or HATU (1-2 eq.), HOBt (l-2eq.) and DIPEA (2-4eq.) were added. In some cases DMF was added as a co-solvent to improve solubility. After stirring at ambient temperature until the reaction was complete (LCMS/TLC), the reaction mixture was concentrated in vacuo. The resultant residue was dissolved in ethyl acetate and washed with water before the organic layer was isolated, dried over sodium sulfate, then concentrated in vacuo and purified by one of the general purification methods described below. If necessary, any protecting groups were then removed using one of the deprotection conditions described below. [00718] Deprotection general methods
[00719] Method A: Aqueous HCl (IN or 2N) was added to a mixture of the protected substrate in an appropriate solvent at ambient temperature. The mixture was stirred until analysis (TLC/LCMS) showed complete consumption of starting material. The reaction mixture was neutralized, concentrated in vacuo and subjected to purification. [00720] Method B: A solution of the substrate in methanol was loaded onto an
Isolute® SCX-2 cartridge. The cartridge was then washed with methanol before the desired product was eluted using 2M ammonia in MeOH and the eluent collected then concentrated to give a residue. The residue was subjected to purification.
[00721] Method C: TBAF in THF was added to a solution of the silyl ether, the mixture stirred at ambient temperature until analysis (TLC/LCMS) showed complete consumption of starting material. The reaction mixture was concentrated in vacuo and subjected to purification.
[00722] Method D: TFA was added to the substrate either neat or as a solution in
DCM. The reaction mixture was stirred at ambient temperature until analysis (TLC/LCMS) showed complete consumption of starting material. The reaction mixture was concentrated in vacuo, and subjected to purification.
[00723] Method E: A 20% solution of piperidine in DME was added to the substrate.
The reaction mixture was stirred at ambient temperature until analysis (TLC/LCMS) showed complete consumption of starting material. The reaction mixture was then concentrated. [00724] Method F: A 4N HCl solution in dioxane was added to the substrate. The reaction mixture was stirred at ambient temperature until analysis (TLC/LCMS) showed complete consumption of starting material. The reaction mixture was then concentrated. [00725] Method G: An aliquot (3 mol equivalents) of freshly prepared HCl in methanol solution [concentrated HCl (0.14 ml) in methanol (25 ml)] was added to the coupled substrate at ambient temperature. The mixture was stirred until analysis (TLC/LCMS) showed complete consumption of starting material. The contents were evaporated to dryness and the residue was dissolved in dichloromethane and treated with triethylamine (3 mol equivalents) at room temperature for 10 min. The mixture was then concentrated in vacuo and the residue subjected to purification.
[00726] Purification general methods
[00727] Method A: Si-SPE or Si-ISCO, ethyl acetate/cyclohexane gradient
[00728] Method B: Si-SPE or Si-ISCO, ethyl acetate/DCM gradient
[00729] Method C: Si-SPE or Si-ISCO, methanol/DCM gradient
[00730] Method D: Si-SPE or Si-ISCO, methanol/ethyl acetate gradient
[00731] Method E: reversed phase HPLC Phenomenex Luna 5 phenyl/hexyl, 0.1%
TFA in water on a gradient of methanol
[00732] Method F: reversed phase HPLC Phenomenex Luna 5 phenyl/hexyl, 0.1%
TFA in water on a gradient of acetonitrile
[00733] Method G: reversed phase HPLC Phenomenex Luna 5 phenyl/hexyl, 0.1%
HCO2H in water on a gradient of methanol
[00734] Method H: reversed phase HPLC Phenomenex Luna 5 phenyl/hexyl, 0.1%
HCO2H in water on a gradient of acetonitrile
[00735] Method I: A solution of the substrate in methanol was loaded onto an
Isolute® SCX-2 cartridge. The cartridge was then washed with methanol before the desired product was eluted using 2M ammonia in MeOH.
[00736] Method J: Si-SPE or Si-ISCO, ethyl acetate/hexane gradient.
[00737] Method K: reversed phase HPLC Sunfϊre Cl 8, 0.05% TFA in water on a gradient of acetonitrile.
[00738] Method L: Si-SPE or Si-ISCO, ethanol/ethyl acetate gradient.
[00739] Method M: Si-SPE, ether/pentane gradient then methanol/ether gradient
[00740] Deviations from general methods:
1 Triturated in hot methanol; 2 recrystallised from ethyl acetate; 3 triturated in diethyl ether; 4 recrystallised from diethyl ether; 5 recrystallised from 5% MeOH in CHCl3; 6 Si-SPE ether/pentane then methanol in ether eluent; 7 triturated in ethyl acetate, 8 ester saponification with lithium hydroxide, 9 A C18 column used; 10 reaction carried out in DMF; n chloroform/methanol recrystallisation; 12 trituration in acetonitrile; 13 DMF used as reaction co-solvent; 14 recrystallisation in methanol; 15 final elution with 10% methanol in ethyl acetate; 16 Reaction mixture heated at 550C; 17 triturated in diethyl ether/DCM.
Figure imgf000164_0001
Figure imgf000165_0001
Figure imgf000166_0001
Figure imgf000167_0001
J =
Figure imgf000168_0001
Figure imgf000169_0001
c]pyridine-2-carboxamide
Figure imgf000170_0001
Figure imgf000171_0001
Figure imgf000172_0001
Figure imgf000173_0001
Figure imgf000174_0001
Figure imgf000175_0001
Figure imgf000176_0001
Figure imgf000177_0001
Figure imgf000178_0001
Figure imgf000179_0001
Figure imgf000180_0001
Figure imgf000181_0001
Figure imgf000182_0001
Figure imgf000183_0001
Figure imgf000184_0001
Figure imgf000185_0001
Figure imgf000186_0001
Figure imgf000187_0001
Figure imgf000188_0001
Figure imgf000189_0001

Claims

We claim:
1. A compound selected from Formula I:
Figure imgf000190_0001
and solvates and salts thereof, wherein:
Z1 Is CR1 Or N;
Z2 is CR2 or N;
Z3 is CR3 or N;
Z4 is CR4 or N; where one or two of Z1, Z2, Z3, and Z4 are N;
R1, R2, R3 and R4 are independently selected from H, halo, CN, CF3, -OCF3, -NO2, -(CR14R15)nC(=Y)Rn, -(CR14R15)nC(=Y)ORn, -(CR14R15)nC(=Y)NRnR12, -(CR14R15)nNRnR12, -(CR14R15)nORn, -(CR14R1 ^nSR1 \ -(CR14R15)nNR12C(=Y)Rn, -(CR14R15)nNR12C(=Y)OR11, -(CR14R15)nNR13C(=Y)NR11R12, -(CR14R15)nNR12SO2R11, -(CR14R1 ^nOCC=Y)R11 , -(CR14R1 ^nOCC=Y)OR11 , -(CR14R1 ^nOCC=Y)NR11R12, -(CR14R15)nOS(O)2(OR11), -(CR14R15)nOP(=Y)(OR11)(OR12), -(CR14R15)nOP(OR11)(OR12), -(CR14R15)nS(O)Rn, -(CR14R15)nS(O)2Rn, -(CR14R15)n S(O)2NR11R12, -(CR14R1^nS(O)(OR11X -(CR14R1^nS(O)2(OR11), -(CR14R15)n SC(=Y)Rn, -(CR14R15)nSC(=Y)ORn, -(CR14R15)nSC(=Y)NRnR12, CJ-CI2 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl;
Figure imgf000190_0002
R5 and R6 are independently selected from H or Ci-Ci2 alkyl;
X1 is selected from R11, -OR11, -NR11R12, -S(O)R11, and -S(O)2R11; when X1 is R11 or -OR11, R11 or -OR11 of X1 and -R5 are optionally taken together with the nitrogen atom to which they are attached to form a 4-7 membered saturated or unsaturated ring having 0-2 additional heteroatoms selected from O, S and N, wherein said ring is optionally substituted with one or more groups selected from halo, CN, CF3, -OCF3, -NO2, oxo, -Si(Ci-C6 alkyl), -(CR19R20)nC(=Y')R16, -(CR19R20)n C(=Y')OR16, -(CR19R20)nC(=Y')NR16R17, -(CR19R20)nNR16R17, -(CR19R20)nOR16, -(CR19R20)n-SR16, -(CR19R20)n NR16C(=Y')R17, -(CR19R20)n NR16C(=Y')OR17, -(CR19R20)n NR18C(=Y')NR16R17, -(CR19R20)nNR17SO2R16, -(CR19R20)nOC(=Y')R16, -(CR19R20)nOC(=Y')OR16, -(CR19R20)nOC(=Y')NR16R17, -(CR19R20)nOS(O)2(OR16), -(CR19R20)nOP(=Y')(OR16)(OR17), -(CR19R20)nOP(OR16)(OR17), -(CR19R20)nS(O)R16, -(CR19R20)nS(O)2R16, -(CR19R20)nS(O)2NR16R17, -(CR19R20)nS(O)(OR16), -(CR19R20X S(O)2(OR16), -(CR19R20)n SC(=Y')R16, -(CR19R20)n SC(=Y')OR16, -(CR19R20X SC(=Y')NR16R17, and R21;
X2 is selected from carbocyclyl, heterocyclyl, aryl, and heteroaryl;
R11, R12 and R13 are independently H, Ci-Ci2 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl, or R11 and R12 together with the nitrogen to which they are attached form a 3-8 membered saturated, unsaturated or aromatic ring having 0-2 heteroatoms selected from O, S and N, wherein said ring is optionally substituted with one or more groups selected from halo, CN, CF3, -OCF3, -NO2, Ci-C6 alkyl, -OH, -SH, -0(Ci-C6 alkyl), -S(Ci-C6 alkyl), -NH2, -NH(Ci-C6 alkyl), -N(Ci-C6 alkyl)2, -SO2(Ci-C6 alkyl), -CO2H, -CO2(Ci-C6 alkyl), -C(O)NH2, -C(O)NH(Ci-C6 alkyl), -C(O)N(Ci-C6 alkyl)2, -N(Ci-C6 alkyl)C(O)(Ci-C6 alkyl), -NHC(O)(Ci-C6 alkyl), -NHSO2(Ci-C6 alkyl), -N(Ci-C6 alkyl)SO2(Ci-C6 alkyl), -SO2NH2, -SO2NH(Ci-C6 alkyl), -SO2N(Ci-C6 alkyl)2, -OC(O)NH2, -OC(O)NH(Ci-C6 alkyl), -OC(O)N(Ci-C6 alkyl)2, -OC(O)O(Ci-C6 alkyl), -NHC(O)NH(Ci-C6 alkyl), -NHC(O)N(Ci-C6 alkyl)2, -N(Ci-C6 alkyl)C(O)NH(Ci-C6 alkyl), -N(Ci-C6 alkyl)C(O)N(Ci- C6 alkyl)2, -NHC(O)NH(Ci-C6 alkyl), -NHC(O)N(Ci-C6 alkyl)2, -NHC(O)O(Ci-C6 alkyl), and -N(Ci-C6 alkyl)C(O)O(Ci-C6 alkyl);
R14 and R15 are independently selected from H, Ci-Ci2 alkyl, aryl, carbocyclyl, heterocyclyl, and heteroaryl; m and n are independently selected from O, 1, 2, 3, 4, 5, or 6;
Y is independently O, NR11, or S; wherein each said alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl and heteroaryl of R1, R2, R3, R4, R5, R6, X1, X2, R11, R12, R13, R14, and R15 is independently optionally substituted with one or more groups independently selected from halo, CN, CF3, -OCF3, -NO2, oxo, -Si(C1-C6 alkyl), -(CR19R20)nC(=Y')R16, -(CR19R20)n C(=Y')OR16, -(CR19R20)nC(=Y')NR16R17, -(CR19R20)nNR16R17, -(CR19R20)nOR16, -(CR19R20)n-SR16, -(CR19R20)n NR16C(=Y')R17, -(CR19R20X NR16C(=Y')OR17, -(CR19R20)n NR18C(=Y')NR16R17, -(CR19R20XNR17SO2R16, -(CR19R20)nOC(=Y')R16, -(CR19R20)nOC(=Y')OR16, -(CR19R20)nOC(=Y')NR16R17, -(CR19R20XOS(O)2(OR16), -(CR19R20)nOP(=Y')(OR16)(OR17), -(CR19R20χθP(OR16)(OR17), -(CR19R20XS(O)R16, -(CR19R20XS(O)2R16, -(CR19R20XS(O)2NR16R17, -(CR19R20)nS(O)(OR16), -(CR19R20X S(O)2(OR16), -(CR19R20X SC(=Y')R16, -(CR19R20X SC(=Y')0R16, -(CR19R20X SC(=Y')NR16R17, and R21; each R16, R17 and R18 is independently H, CJ-CI2 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl, wherein said alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, or heteroaryl is optionally substituted with one or more groups selected from halo, CN, -OCF3, CF3, -NO2, Ci-C6 alkyl, -OH, -SH, -0(Ci-C6 alkyl), -S(Ci-C6 alkyl), -NH2, -NH(Ci-C6 alkyl), -N(Ci-C6 alkyl)2, -SO2(Ci-C6 alkyl), -CO2H, -CO2(Ci-C6 alkyl), -C(O)NH2, -C(O)NH(Ci-C6 alkyl), -C(O)N(Ci-C6 alkyl)2, -N(Ci-C6 alkyl)C(O)(Ci-C6 alkyl), -NHC(O)(Ci-C6 alkyl), -NHSO2(Ci-C6 alkyl), -N(Ci-C6 alkyl)SO2(Ci-C6 alkyl), -SO2NH2, -SO2NH(Ci-C6 alkyl), -SO2N(Ci-C6 alkyl)2, -OC(O)NH2, -OC(O)NH(Ci-C6 alkyl), -OC(O)N(Ci-C6 alkyl)2, -OC(O)O(Ci-C6 alkyl), -NHC(O)NH(Ci-C6 alkyl), -NHC(O)N(Ci-C6 alkyl)2, -N(Ci-C6 alkyl)C(O)NH(Ci-C6 alkyl), -N(Ci-C6 alkyl)C(O)N(Ci-C6 alkyl)2, -NHC(O)NH(Ci-C6 alkyl), -NHC(O)N(Ci-C6 alkyl)2, -NHC(O)O(Ci-C6 alkyl), and -N(Ci-C6 alkyl)C(O)O(Ci-C6 alkyl); or R16 and R17 together with the nitrogen to which they are attached form a 3-8 membered saturated, unsaturated or aromatic ring having 0-2 heteroatoms selected from O, S and N, wherein said ring is optionally substituted with one or more groups selected from halo, CN, -OCF3, CF3, -NO2, Ci-C6 alkyl, -OH, -SH, -0(Ci-C6 alkyl), -S(Ci-C6 alkyl), -NH2, -NH(Ci-C6 alkyl), -N(Ci -C6 alkyl)2, -SO2(Ci-C6 alkyl), -CO2H, -CO2(Ci-C6 alkyl), -C(O)NH2, -C(O)NH(Ci-C6 alkyl), -C(O)N(Ci-C6 alkyl)2, -N(Ci-C6 alkyl)C(O)(Ci-C6 alkyl), -NHC(O)(Ci-C6 alkyl), -NHSO2(Ci-C6 alkyl), -N(Ci-C6 alkyl)SO2(Ci-C6 alkyl), -SO2NH2, -SO2NH(Ci-C6 alkyl), -SO2N(Ci-C6 alkyl)2, -OC(O)NH2, -OC(O)NH(Ci-C6 alkyl), -OC(O)N(CrC6 alkyl)2, -OC(O)O(CrC6 alkyl), -NHC(0)NH(CrC6 alkyl), -NHC(O)N(Ci-C6 alkyl)2, -N(Ci-C6 alkyl)C(O)NH(Ci-C6 alkyl), -N(Ci-C6 alkyl)C(O)N(Ci-
C6 alkyl)2, -NHC(O)NH(Ci-C6 alkyl), -NHC(O)N(Ci-C6 alkyl)2, -NHC(O)O(Ci-C6 alkyl), and -N(C1-C6 alkyl)C(O)O(CrC6 alkyl);
R19 and R20 are independently selected from H, Ci-Ci2 alkyl, -(CH2)n-aryl, -(CH2)n- carbocyclyl, -(CH2)n-heterocyclyl, and -(CH2)n-heteroaryl;
R21 is Ci-Ci2 alkyl, C2-Cs alkenyl, C2-Cs alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl, wherein each member of R21 is optionally substituted with one or more groups selected from halo, oxo, CN, -OCF3, CF3, -NO2, C1-C6 alkyl, -OH, -SH, -0(C1-C6 alkyl), -S(Ci-C6 alkyl), -NH2, -NH(Ci-C6 alkyl), -N(Ci-C6 alkyl)2, -SO2(Ci-C6 alkyl), -CO2H, -CO2(Ci-C6 alkyl), -C(O)NH2, -C(O)NH(Ci-C6 alkyl), -C(O)N(Ci-C6 alkyl)2, -N(Ci-C6 alkyl)C(O)(Ci-C6 alkyl), -NHC(0)(CrC6 alkyl), -NHSO2(Ci-C6 alkyl), -N(Ci -C6 alkyl)SO2(Ci-C6 alkyl), -SO2NH2, -SO2NH(Ci-C6 alkyl), -SO2N(Ci-C6 alkyl)2, -OC(O)NH2, -OC(O)NH(Ci-C6 alkyl), -OC(O)N(Ci-C6 alkyl)2, -OC(O)O(Ci-C6 alkyl), -NHC(O)NH(Ci-C6 alkyl), -NHC(O)N(Ci-C6 alkyl)2, -N(Ci-C6 alkyl)C(O)NH(Ci-C6 alkyl), -N(Ci-C6 alkyl)C(O)N(Ci-C6 alkyl)2, -NHC(0)NH(CrC6 alkyl), -NHC(0)N(CrC6 alkyl)2, -NHC(O)O(Ci-C6 alkyl), and -N(Ci-C6 alkyl)C(O)O(Ci-C6 alkyl); each Y' is independently O, NR22, or S; and
R22 is H or Ci-Ci2 alkyl.
2. The compound of claim 1 selected from Formulae I-b, I-f, I-g, and I-h:
Figure imgf000193_0001
3. The compound of claim 2 wherein X1 is selected from:
Figure imgf000194_0001
I-UINL
OK HO' V HO Y 'O' O *
OH
Figure imgf000194_0002
.x- x
O Ό V- ^. ^.
O O
4. The compound of claim 2 wherein X1 is selected from:
H< X HOV
HO HO
Figure imgf000194_0003
5. The compound of claim 2 wherein X1 is R11, and R11 and R5 are taken together with the nitrogen atom to which they are attached to form:
Figure imgf000194_0004
6. The compound of claim 2 wherein X is:
Figure imgf000195_0001
7. The compound of claim 2 wherein R1 is selected from H, CH3, CF3, CN, -NR11R12, -OR11, and Cl.
8. The compound of claim 2 wherein R is selected from H, CH3, F, or CF3.
9. The compound of claim 2 wherein R4 is selected from CF3, Br, Cl, CN, -NR11R12, -OR11, and -CC=O)NR11R12.
10. The compound of claim 9 wherein R4 is selected from Cl, Br, Me, Et, F, CHF2, CF3 or -OH.
11. The compound of claim 2 wherein R5 is H or methyl.
12. The compound of claim 2 wherein R6 is H or methyl.
13. The compound of claim 1 wherein W is OR11.
14. The compound of claim 13 wherein W is OH.
15. A compound selected from the title compounds in Examples 5-19, 97-109 and 138-139, and Examples 20-96 and 111-160 in Tables 2, 3 and 4.
16. A pharmaceutical composition comprising a compound of any one of claims 1-15, and a pharmaceutically acceptable carrier.
17. The pharmaceutical composition of claim 16, further comprising a second chemotherapeutic agent.
18. The pharmaceutical composition of claim 16, further comprising a second anti-inflammatory agent.
19. A method of inhibiting abnormal cell growth or treating a hyperproliferative disorder in a mammal comprising administering to said mammal a therapeutically effective amount of a pharmaceutical composition of claim 16 or 17.
20. A method of treating an inflammatory disease in a mammal comprising administering to said mammal a therapeutically effective amount of a pharmaceutical composition of claim 16 or 18.
21. The method of claim 19 or 20, wherein said second chemotherapeutic or antiinflammatory agent is administered to said mammal sequentially or consecutively.
22. A method of treating an autoimmune disease, destructive bone disorder, proliferative disorders, infectious disease, viral disease, fϊbrotic disease, neurodegenerative disease, pancreatitis or kidney disease in a mammal comprising administering to said mammal a therapeutically effective amount of a pharmaceutical composition of claim 16.
23. The method of claim 22, further comprising administering to said mammal a second therapeutic agent, wherein said second agent is administered to said mammal sequentially or consecutively.
PCT/US2007/076344 2006-08-21 2007-08-20 Aza-benzofuranyl compounds and methods of use WO2008024725A1 (en)

Priority Applications (12)

Application Number Priority Date Filing Date Title
JP2009525720A JP5448818B2 (en) 2006-08-21 2007-08-20 Azabenzofuranyl compounds and methods of use
ES07814272T ES2376771T3 (en) 2006-08-21 2007-08-20 Aza-benzofuranyl compounds and methods of use
EP07814272A EP2069354B1 (en) 2006-08-21 2007-08-20 Aza-benzofuranyl compounds and methods of use
CA002660546A CA2660546A1 (en) 2006-08-21 2007-08-20 Aza-benzofuranyl compounds and methods of use
BRPI0714635-3A BRPI0714635A2 (en) 2006-08-21 2007-08-20 compounds, pharmaceutical composition, method for inhibiting abnormal cell growth or treating hyperproliferative dysfunction, method for treating an inflammatory disease, and method for treating an autoimmune disease
AT07814272T ATE531720T1 (en) 2006-08-21 2007-08-20 AZA-BENZOFURANYL COMPOUNDS AND METHODS OF USE THEREOF
AU2007286808A AU2007286808B2 (en) 2006-08-21 2007-08-20 Aza-benzofuranyl compounds and methods of use
KR1020097005786A KR101428116B1 (en) 2006-08-21 2007-08-20 Aza―benzofuranyl compounds and methods of use
CN200780039152.0A CN101605794B (en) 2006-08-21 2007-08-20 Aza-benzofuranyl compounds and methods of use
MX2009001878A MX2009001878A (en) 2006-08-21 2007-08-20 Aza-benzofuranyl compounds and methods of use.
IL196956A IL196956A (en) 2006-08-21 2009-02-09 3-phenylamino-furo[3,2-c]pyridine-2-carbonic acid, esters and amides thereof and pharmaceutical compositions comprising the same for treating inflammatory and hyperproliferative diseases
NO20091197A NO20091197L (en) 2006-08-21 2009-03-20 Azabenzofuranyl compounds and uses thereof

Applications Claiming Priority (8)

Application Number Priority Date Filing Date Title
US83916106P 2006-08-21 2006-08-21
US60/839,161 2006-08-21
US87159106P 2006-12-22 2006-12-22
US60/871,591 2006-12-22
US91762307P 2007-05-11 2007-05-11
US60/917,623 2007-05-11
US94474107P 2007-06-18 2007-06-18
US60/944,741 2007-06-18

Publications (1)

Publication Number Publication Date
WO2008024725A1 true WO2008024725A1 (en) 2008-02-28

Family

ID=38925647

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2007/076344 WO2008024725A1 (en) 2006-08-21 2007-08-20 Aza-benzofuranyl compounds and methods of use

Country Status (16)

Country Link
US (1) US20080085886A1 (en)
EP (1) EP2069354B1 (en)
JP (1) JP5448818B2 (en)
KR (1) KR101428116B1 (en)
AR (1) AR062468A1 (en)
AT (1) ATE531720T1 (en)
AU (1) AU2007286808B2 (en)
BR (1) BRPI0714635A2 (en)
CA (1) CA2660546A1 (en)
ES (1) ES2376771T3 (en)
IL (1) IL196956A (en)
MX (1) MX2009001878A (en)
NO (1) NO20091197L (en)
PE (1) PE20080611A1 (en)
TW (1) TWI411614B (en)
WO (1) WO2008024725A1 (en)

Cited By (48)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010068738A1 (en) 2008-12-10 2010-06-17 Dana-Farber Cancer Institute, Inc. Mek mutations conferring resistance to mek inhibitors
WO2010119906A1 (en) 2009-04-14 2010-10-21 日産化学工業株式会社 Haloalkylsulfonanilide derivative
WO2010145197A1 (en) * 2009-06-15 2010-12-23 Chemizon (Beijing), Ltd. Novel 6-arylamino pyridone sulfonamides and 6-arylamino pyrazinone sulfonamdies as mek inhibitors
WO2011106298A1 (en) 2010-02-25 2011-09-01 Dana-Farber Cancer Institute, Inc. Braf mutations conferring resistance to braf inhibitors
WO2012071458A1 (en) 2010-11-22 2012-05-31 Array Biopharma Inc. Heterocyclic inhibitors of beta - secretase for the treatment of neurodegenerative diseases
WO2013019906A1 (en) 2011-08-01 2013-02-07 Genentech, Inc. Methods of treating cancer using pd-1 axis binding antagonists and mek inhibitors
WO2013169858A1 (en) 2012-05-08 2013-11-14 The Broad Institute, Inc. Diagnostic and treatment methods in patients having or at risk of developing resistance to cancer therapy
WO2014071183A1 (en) 2012-11-02 2014-05-08 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Method of reducing adverse effects in a cancer patient undergoing treatment with a mek inhibitor
US9125937B2 (en) 2008-01-31 2015-09-08 Institut National De La Sante Et De La Recherche Medicale (Inserm) Labelled analogues of halobenzamides as multimodal radiopharmaceuticals and their precursors
WO2015138920A1 (en) 2014-03-14 2015-09-17 Novartis Ag Antibody molecules to lag-3 and uses thereof
WO2016011160A1 (en) 2014-07-15 2016-01-21 Genentech, Inc. Compositions for treating cancer using pd-1 axis binding antagonists and mek inhibitors
WO2016040880A1 (en) 2014-09-13 2016-03-17 Novartis Ag Combination therapies of alk inhibitors
WO2016054555A2 (en) 2014-10-03 2016-04-07 Novartis Ag Combination therapies
WO2016061142A1 (en) 2014-10-14 2016-04-21 Novartis Ag Antibody molecules to pd-l1 and uses thereof
US9328066B2 (en) 2011-05-27 2016-05-03 Bayer Intellectual Property Gmbh Chiral synthesis of N-{3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]-6-methoxyphenyl}-1-[2,3-dihydroxy-propyl]cyclopropanesulfonamides
WO2016100882A1 (en) 2014-12-19 2016-06-23 Novartis Ag Combination therapies
US9532987B2 (en) 2013-09-05 2017-01-03 Genentech, Inc. Use of a combination of a MEK inhibitor and an ERK inhibitor for treatment of hyperproliferative diseases
WO2017019894A1 (en) 2015-07-29 2017-02-02 Novartis Ag Combination therapies comprising antibody molecules to lag-3
WO2017019897A1 (en) 2015-07-29 2017-02-02 Novartis Ag Combination therapies comprising antibody molecules to tim-3
WO2017106656A1 (en) 2015-12-17 2017-06-22 Novartis Ag Antibody molecules to pd-1 and uses thereof
WO2017180581A1 (en) 2016-04-15 2017-10-19 Genentech, Inc. Diagnostic and therapeutic methods for cancer
US10047066B2 (en) 2007-11-30 2018-08-14 Newlink Genetics Corporation IDO inhibitors
WO2018213807A1 (en) * 2017-05-19 2018-11-22 Nflection Therapeutics, Inc. Fused heteroaromatic-aniline compounds for treatment of dermal disorders
WO2019051296A1 (en) 2017-09-08 2019-03-14 Genentech, Inc. Diagnostic and therapeutic methods for cancer
WO2019099838A1 (en) 2017-11-16 2019-05-23 Novartis Ag Combination therapies
EP3514179A1 (en) 2014-01-24 2019-07-24 Dana-Farber Cancer Institute, Inc. Antibody molecules to pd-1 and uses thereof
WO2019158579A1 (en) 2018-02-13 2019-08-22 Vib Vzw Targeting minimal residual disease in cancer with rxr antagonists
WO2019224718A2 (en) 2018-05-24 2019-11-28 Janssen Biotech, Inc. Psma binding agents and uses thereof
WO2020012337A1 (en) 2018-07-10 2020-01-16 Novartis Ag 3-(5-amino-1-oxoisoindolin-2-yl)piperidine-2,6-dione derivatives and their use in the treatment of i karos family zinc finger 2 (ikzf2)-dependent diseases
WO2020128972A1 (en) 2018-12-20 2020-06-25 Novartis Ag Dosing regimen and pharmaceutical combination comprising 3-(1-oxoisoindolin-2-yl)piperidine-2,6-dione derivatives
WO2020165833A1 (en) 2019-02-15 2020-08-20 Novartis Ag 3-(1-oxo-5-(piperidin-4-yl)isoindolin-2-yl)piperidine-2,6-dione derivatives and uses thereof
WO2020165834A1 (en) 2019-02-15 2020-08-20 Novartis Ag Substituted 3-(1-oxoisoindolin-2-yl)piperidine-2,6-dione derivatives and uses thereof
US10894830B2 (en) 2015-11-03 2021-01-19 Janssen Biotech, Inc. Antibodies specifically binding PD-1, TIM-3 or PD-1 and TIM-3 and their uses
WO2021123996A1 (en) 2019-12-20 2021-06-24 Novartis Ag Uses of anti-tgf-beta antibodies and checkpoint inhibitors for the treatment of proliferative diseases
US11078540B2 (en) 2010-03-09 2021-08-03 Dana-Farber Cancer Institute, Inc. Methods of diagnosing and treating cancer in patients having or developing resistance to a first cancer therapy
US11161845B2 (en) 2017-05-19 2021-11-02 Nflection Therapeutics, Inc. Pyrrolopyridine-aniline compounds for treatment of dermal disorders
WO2021260528A1 (en) 2020-06-23 2021-12-30 Novartis Ag Dosing regimen comprising 3-(1-oxoisoindolin-2-yl)piperidine-2,6-dione derivatives
WO2022162518A2 (en) 2021-01-28 2022-08-04 Janssen Biotech, Inc. Psma binding proteins and uses thereof
WO2022169780A1 (en) 2021-02-02 2022-08-11 Les Laboratoires Servier Selective bcl-xl protac compounds and methods of use
WO2022195551A1 (en) 2021-03-18 2022-09-22 Novartis Ag Biomarkers for cancer and methods of use thereof
WO2022215011A1 (en) 2021-04-07 2022-10-13 Novartis Ag USES OF ANTI-TGFβ ANTIBODIES AND OTHER THERAPEUTIC AGENTS FOR THE TREATMENT OF PROLIFERATIVE DISEASES
WO2022243846A1 (en) 2021-05-18 2022-11-24 Novartis Ag Combination therapies
US11572344B2 (en) 2018-11-20 2023-02-07 Nflection Therapeutics, Inc. Cyanoaryl-aniline compounds for treatment of dermal disorders
US11591395B2 (en) 2019-04-19 2023-02-28 Janssen Biotech, Inc. Methods of treating prostate cancer with an anti-PSMA/CD3 antibody
WO2023225320A1 (en) 2022-05-20 2023-11-23 Novartis Ag Epha2 bcl-xl inhibitor antibody-drug conjugates and methods of use thereof
WO2023225336A1 (en) 2022-05-20 2023-11-23 Novartis Ag Met bcl-xl inhibitor antibody-drug conjugates and methods of use thereof
WO2024033381A1 (en) 2022-08-10 2024-02-15 Vib Vzw Inhibition of tcf4/itf2 in the treatment of cancer
EP4324518A2 (en) 2014-01-31 2024-02-21 Novartis AG Antibody molecules to tim-3 and uses thereof

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ES2528797T3 (en) * 2006-08-21 2015-02-12 Genentech, Inc. Aza-benzothiophenyl compounds and methods of use
CN106883270B (en) 2015-11-26 2019-03-26 财团法人工业技术研究院 Organometallic compound and organic light-emitting device including the same

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6653346B1 (en) * 2002-02-07 2003-11-25 Galileo Pharmaceuticals, Inc. Cytoprotective benzofuran derivatives
WO2005047292A1 (en) * 2003-11-04 2005-05-26 Merck Patent Gmbh Use of thienopyrimidines
WO2005061476A2 (en) * 2003-12-22 2005-07-07 Basilea Pharmaceutica Ag Aroylfuranes and aroylthiophenes
EP1582521A1 (en) * 2003-01-09 2005-10-05 Tanabe Seiyaku Co., Ltd. Fused furan compound
WO2007027855A2 (en) * 2005-09-01 2007-03-08 Array Biopharma Inc. Raf inhibitor compounds and methods of use thereof

Family Cites Families (24)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5328920A (en) * 1991-04-17 1994-07-12 Hoechst-Roussel Pharmaceuticals Incorporated Substituted (pyridinylamino)-indoles
US5214204A (en) * 1991-07-19 1993-05-25 Abbott Laboratories Arylamidoalkyl-n-hydroxyurea compounds having lipoxygenase inhibitory activity
DE69216509T2 (en) * 1991-08-12 1997-05-28 Takeda Chemical Industries Ltd Condensed pyrimidine derivatives, their preparation and their use as anti-tumor agents
EP0545478A1 (en) * 1991-12-03 1993-06-09 MERCK SHARP &amp; DOHME LTD. Heterocyclic compounds as tachykinin antagonists
JPH05345780A (en) * 1991-12-24 1993-12-27 Kumiai Chem Ind Co Ltd Pyrimidine or triazine derivative and herbicide
US5852046A (en) * 1993-08-03 1998-12-22 Hoechst Aktiengesellschaft Benzo-fused heterocyclic compounds having a 5-membered ring processes for their preparation their use as medicaments their use as diagnostic agents and medicaments containing them
US5350748A (en) * 1993-08-18 1994-09-27 Warner-Lambert Company 3-thio or amino substituted-benzo[b]thiophene-2-carboxamides and 3-oxygen, thio, or amino substituted-benzofuran-2-carboxamides as inhibitors of cell adhesion
DE4425609A1 (en) * 1994-07-20 1996-01-25 Bayer Ag Benzofuranyl and Benzothienyloxazolidinone
FR2777886B1 (en) * 1998-04-27 2002-05-31 Adir NOVEL BENZOTHIOPHENIC, BENZOFURANIC AND INDOLIC DERIVATIVES, PROCESS FOR THEIR PREPARATION AND THE PHARMACEUTICAL COMPOSITIONS CONTAINING THEM
US6410562B1 (en) * 1998-12-18 2002-06-25 Eli Lilly And Company Hypoglycemic imidazoline compounds
JP2000204079A (en) * 1999-01-13 2000-07-25 Warner Lambert Co Diarylamine
TR200102030T2 (en) * 1999-01-13 2002-01-21 Warner-Lambert Company pharmaceutical composition including on
FR2799756B1 (en) * 1999-10-15 2001-12-14 Adir NOVEL BENZOTHIOPHENIC, BENZOFURANIC AND INDOLIC DERIVATIVES, PROCESS FOR THEIR PREPARATION AND THE PHARMACEUTICAL COMPOSITIONS CONTAINING THEM
PL203387B1 (en) * 2000-07-19 2009-09-30 Warner Lambert Co Oxygenated esters of 4-iodo phenylamino benzhydroxamic acids
DE10110750A1 (en) * 2001-03-07 2002-09-12 Bayer Ag Novel aminodicarboxylic acid derivatives with pharmaceutical properties
AR038971A1 (en) * 2002-03-13 2005-02-02 Array Biopharma Inc DERIVATIVES OF BENCIMIDAZOL N3 RENTED AS MEK INHIBITORS
PL378635A1 (en) * 2002-03-13 2006-05-15 Array Biopharma, Inc. N3 alkylated benzimidazole derivatives as mek inhibitors
US20040009976A1 (en) * 2002-04-30 2004-01-15 Kumiko Takeuchi Hypoglycemic imidazoline compounds
CL2003002287A1 (en) * 2002-11-25 2005-01-14 Wyeth Corp COMPOUNDS DERIVED FROM TIENO [3,2-b] -PIRIDINA-6-CARBONITRILOS AND TIENEO [2,3-b] -PIRIDINA-5-CARBONITRILS, PHARMACEUTICAL COMPOSITION, PROCEDURE OF PREPARATION AND INTERMEDIARY COMPOUNDS, AND THEIR USE IN THE TREATMENT OF CANCER, APOPLEJIA, OSTEOPOROSIS
US7144907B2 (en) * 2003-09-03 2006-12-05 Array Biopharma Inc. Heterocyclic inhibitors of MEK and methods of use thereof
WO2006106326A1 (en) * 2005-04-06 2006-10-12 Astrazeneca Ab Substituted heterocycles and their use as chk1, pdk1 and pak inhibitors
TWI389895B (en) * 2006-08-21 2013-03-21 Infinity Discovery Inc Compounds and methods for inhibiting the interaction of bcl proteins with binding partners
ES2528797T3 (en) * 2006-08-21 2015-02-12 Genentech, Inc. Aza-benzothiophenyl compounds and methods of use
CA2662348A1 (en) * 2006-09-21 2008-03-27 Wyeth Indolylalkylpyridin-2-amines for the inhibition of .beta.-secretase

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6653346B1 (en) * 2002-02-07 2003-11-25 Galileo Pharmaceuticals, Inc. Cytoprotective benzofuran derivatives
EP1582521A1 (en) * 2003-01-09 2005-10-05 Tanabe Seiyaku Co., Ltd. Fused furan compound
WO2005047292A1 (en) * 2003-11-04 2005-05-26 Merck Patent Gmbh Use of thienopyrimidines
WO2005061476A2 (en) * 2003-12-22 2005-07-07 Basilea Pharmaceutica Ag Aroylfuranes and aroylthiophenes
WO2007027855A2 (en) * 2005-09-01 2007-03-08 Array Biopharma Inc. Raf inhibitor compounds and methods of use thereof

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
DAVIES, H. ET AL., NATURE, vol. 417, 2002, pages 949 - 954
HOSHINO, R. ET AL., ONCOGENE, vol. 18, 1999, pages 813 - 822
KOHL ET AL., SCIENCE, vol. 260, 1993, pages 1834 - 1837
MILELLA ET AL., J CLIN INVEST, vol. 108, no. 6, 2001, pages 851 - 859
SEBOLT-LEOPOLD ET AL., NATURE-MEDICINE, vol. 5, no. 7, 1999, pages 810 - 816
TRACHET ET AL., AACR, 6 April 2006 (2006-04-06)

Cited By (70)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10047066B2 (en) 2007-11-30 2018-08-14 Newlink Genetics Corporation IDO inhibitors
EP2252605B1 (en) * 2008-01-31 2018-06-13 Institut National de la Santé et de la Recherche Médicale (INSERM) Labelled analogues of halobenzamides as multimodal radiopharmaceuticals and their precursors
US9125937B2 (en) 2008-01-31 2015-09-08 Institut National De La Sante Et De La Recherche Medicale (Inserm) Labelled analogues of halobenzamides as multimodal radiopharmaceuticals and their precursors
US9084781B2 (en) 2008-12-10 2015-07-21 Novartis Ag MEK mutations conferring resistance to MEK inhibitors
WO2010068738A1 (en) 2008-12-10 2010-06-17 Dana-Farber Cancer Institute, Inc. Mek mutations conferring resistance to mek inhibitors
WO2010119906A1 (en) 2009-04-14 2010-10-21 日産化学工業株式会社 Haloalkylsulfonanilide derivative
WO2010145197A1 (en) * 2009-06-15 2010-12-23 Chemizon (Beijing), Ltd. Novel 6-arylamino pyridone sulfonamides and 6-arylamino pyrazinone sulfonamdies as mek inhibitors
WO2011106298A1 (en) 2010-02-25 2011-09-01 Dana-Farber Cancer Institute, Inc. Braf mutations conferring resistance to braf inhibitors
EP3028699A1 (en) 2010-02-25 2016-06-08 Dana-Farber Cancer Institute, Inc. Braf mutations conferring resistance to braf inhibitors
US9279144B2 (en) 2010-02-25 2016-03-08 Dana-Farber Cancer Institute, Inc. Screening method for BRAF inhibitors
US11078540B2 (en) 2010-03-09 2021-08-03 Dana-Farber Cancer Institute, Inc. Methods of diagnosing and treating cancer in patients having or developing resistance to a first cancer therapy
WO2012071458A1 (en) 2010-11-22 2012-05-31 Array Biopharma Inc. Heterocyclic inhibitors of beta - secretase for the treatment of neurodegenerative diseases
US9328066B2 (en) 2011-05-27 2016-05-03 Bayer Intellectual Property Gmbh Chiral synthesis of N-{3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]-6-methoxyphenyl}-1-[2,3-dihydroxy-propyl]cyclopropanesulfonamides
US10646567B2 (en) 2011-08-01 2020-05-12 Genentech, Inc. Methods of treating cancer using PD-1 axis binding antagonists and MEK inhibitors
US9724413B2 (en) 2011-08-01 2017-08-08 Genentech, Inc. Methods of treating cancer using PD-1 axis binding antagonists and MEK inhibitors
WO2013019906A1 (en) 2011-08-01 2013-02-07 Genentech, Inc. Methods of treating cancer using pd-1 axis binding antagonists and mek inhibitors
WO2013169858A1 (en) 2012-05-08 2013-11-14 The Broad Institute, Inc. Diagnostic and treatment methods in patients having or at risk of developing resistance to cancer therapy
WO2014071183A1 (en) 2012-11-02 2014-05-08 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Method of reducing adverse effects in a cancer patient undergoing treatment with a mek inhibitor
US9532987B2 (en) 2013-09-05 2017-01-03 Genentech, Inc. Use of a combination of a MEK inhibitor and an ERK inhibitor for treatment of hyperproliferative diseases
EP3514179A1 (en) 2014-01-24 2019-07-24 Dana-Farber Cancer Institute, Inc. Antibody molecules to pd-1 and uses thereof
EP4324518A2 (en) 2014-01-31 2024-02-21 Novartis AG Antibody molecules to tim-3 and uses thereof
WO2015138920A1 (en) 2014-03-14 2015-09-17 Novartis Ag Antibody molecules to lag-3 and uses thereof
EP3660050A1 (en) 2014-03-14 2020-06-03 Novartis AG Antibody molecules to lag-3 and uses thereof
US10946093B2 (en) 2014-07-15 2021-03-16 Genentech, Inc. Methods of treating cancer using PD-1 axis binding antagonists and MEK inhibitors
WO2016011160A1 (en) 2014-07-15 2016-01-21 Genentech, Inc. Compositions for treating cancer using pd-1 axis binding antagonists and mek inhibitors
EP3563870A1 (en) 2014-07-15 2019-11-06 F. Hoffmann-La Roche AG Methods of treating cancer using pd-1 axis binding antagonists and mek inhibitors
EP3925622A1 (en) 2014-09-13 2021-12-22 Novartis AG Combination therapies
EP3659621A1 (en) 2014-09-13 2020-06-03 Novartis AG Combination therapies for cancer
WO2016040880A1 (en) 2014-09-13 2016-03-17 Novartis Ag Combination therapies of alk inhibitors
WO2016040892A1 (en) 2014-09-13 2016-03-17 Novartis Ag Combination therapies
EP3662903A2 (en) 2014-10-03 2020-06-10 Novartis AG Combination therapies
WO2016054555A2 (en) 2014-10-03 2016-04-07 Novartis Ag Combination therapies
WO2016061142A1 (en) 2014-10-14 2016-04-21 Novartis Ag Antibody molecules to pd-l1 and uses thereof
EP4245376A2 (en) 2014-10-14 2023-09-20 Novartis AG Antibody molecules to pd-l1 and uses thereof
WO2016100882A1 (en) 2014-12-19 2016-06-23 Novartis Ag Combination therapies
EP3964528A1 (en) 2015-07-29 2022-03-09 Novartis AG Combination therapies comprising antibody molecules to lag-3
EP3878465A1 (en) 2015-07-29 2021-09-15 Novartis AG Combination therapies comprising antibody molecules to tim-3
WO2017019897A1 (en) 2015-07-29 2017-02-02 Novartis Ag Combination therapies comprising antibody molecules to tim-3
WO2017019894A1 (en) 2015-07-29 2017-02-02 Novartis Ag Combination therapies comprising antibody molecules to lag-3
US10894830B2 (en) 2015-11-03 2021-01-19 Janssen Biotech, Inc. Antibodies specifically binding PD-1, TIM-3 or PD-1 and TIM-3 and their uses
WO2017106656A1 (en) 2015-12-17 2017-06-22 Novartis Ag Antibody molecules to pd-1 and uses thereof
WO2017180581A1 (en) 2016-04-15 2017-10-19 Genentech, Inc. Diagnostic and therapeutic methods for cancer
US11161845B2 (en) 2017-05-19 2021-11-02 Nflection Therapeutics, Inc. Pyrrolopyridine-aniline compounds for treatment of dermal disorders
WO2018213807A1 (en) * 2017-05-19 2018-11-22 Nflection Therapeutics, Inc. Fused heteroaromatic-aniline compounds for treatment of dermal disorders
US10988483B2 (en) 2017-05-19 2021-04-27 Nflection Therapeutics, Inc. Fused heteroaromatic-aniline compounds for treatment of dermal disorders
CN111093658A (en) * 2017-05-19 2020-05-01 恩福莱克逊治疗有限公司 Fused heteroaromatic-aniline compounds for the treatment of skin diseases
US11542271B2 (en) 2017-05-19 2023-01-03 Nflection Therapeutics, Inc. Fused heteroaromatic-aniline compounds for treatment of dermal disorders
CN111093658B (en) * 2017-05-19 2022-10-14 恩福莱克逊治疗有限公司 Fused heteroaromatic-aniline compounds for the treatment of skin diseases
AU2018270292B2 (en) * 2017-05-19 2022-02-10 Nflection Therapeutics, Inc. Fused heteroaromatic-aniline compounds for treatment of dermal disorders
WO2019051296A1 (en) 2017-09-08 2019-03-14 Genentech, Inc. Diagnostic and therapeutic methods for cancer
WO2019099838A1 (en) 2017-11-16 2019-05-23 Novartis Ag Combination therapies
WO2019158579A1 (en) 2018-02-13 2019-08-22 Vib Vzw Targeting minimal residual disease in cancer with rxr antagonists
US11746157B2 (en) 2018-05-24 2023-09-05 Janssen Biotech, Inc. PSMA binding agents and uses thereof
WO2019224718A2 (en) 2018-05-24 2019-11-28 Janssen Biotech, Inc. Psma binding agents and uses thereof
WO2020012337A1 (en) 2018-07-10 2020-01-16 Novartis Ag 3-(5-amino-1-oxoisoindolin-2-yl)piperidine-2,6-dione derivatives and their use in the treatment of i karos family zinc finger 2 (ikzf2)-dependent diseases
US11572344B2 (en) 2018-11-20 2023-02-07 Nflection Therapeutics, Inc. Cyanoaryl-aniline compounds for treatment of dermal disorders
WO2020128972A1 (en) 2018-12-20 2020-06-25 Novartis Ag Dosing regimen and pharmaceutical combination comprising 3-(1-oxoisoindolin-2-yl)piperidine-2,6-dione derivatives
WO2020165834A1 (en) 2019-02-15 2020-08-20 Novartis Ag Substituted 3-(1-oxoisoindolin-2-yl)piperidine-2,6-dione derivatives and uses thereof
WO2020165833A1 (en) 2019-02-15 2020-08-20 Novartis Ag 3-(1-oxo-5-(piperidin-4-yl)isoindolin-2-yl)piperidine-2,6-dione derivatives and uses thereof
US11591395B2 (en) 2019-04-19 2023-02-28 Janssen Biotech, Inc. Methods of treating prostate cancer with an anti-PSMA/CD3 antibody
WO2021123996A1 (en) 2019-12-20 2021-06-24 Novartis Ag Uses of anti-tgf-beta antibodies and checkpoint inhibitors for the treatment of proliferative diseases
WO2021260528A1 (en) 2020-06-23 2021-12-30 Novartis Ag Dosing regimen comprising 3-(1-oxoisoindolin-2-yl)piperidine-2,6-dione derivatives
WO2022162518A2 (en) 2021-01-28 2022-08-04 Janssen Biotech, Inc. Psma binding proteins and uses thereof
WO2022169780A1 (en) 2021-02-02 2022-08-11 Les Laboratoires Servier Selective bcl-xl protac compounds and methods of use
WO2022195551A1 (en) 2021-03-18 2022-09-22 Novartis Ag Biomarkers for cancer and methods of use thereof
WO2022215011A1 (en) 2021-04-07 2022-10-13 Novartis Ag USES OF ANTI-TGFβ ANTIBODIES AND OTHER THERAPEUTIC AGENTS FOR THE TREATMENT OF PROLIFERATIVE DISEASES
WO2022243846A1 (en) 2021-05-18 2022-11-24 Novartis Ag Combination therapies
WO2023225320A1 (en) 2022-05-20 2023-11-23 Novartis Ag Epha2 bcl-xl inhibitor antibody-drug conjugates and methods of use thereof
WO2023225336A1 (en) 2022-05-20 2023-11-23 Novartis Ag Met bcl-xl inhibitor antibody-drug conjugates and methods of use thereof
WO2024033381A1 (en) 2022-08-10 2024-02-15 Vib Vzw Inhibition of tcf4/itf2 in the treatment of cancer

Also Published As

Publication number Publication date
ES2376771T3 (en) 2012-03-16
ATE531720T1 (en) 2011-11-15
EP2069354B1 (en) 2011-11-02
IL196956A (en) 2014-04-30
BRPI0714635A2 (en) 2013-06-18
JP2010501585A (en) 2010-01-21
KR101428116B1 (en) 2014-08-07
NO20091197L (en) 2009-05-20
IL196956A0 (en) 2009-11-18
KR20090042332A (en) 2009-04-29
AU2007286808B2 (en) 2012-12-06
US20080085886A1 (en) 2008-04-10
TW200817415A (en) 2008-04-16
CA2660546A1 (en) 2008-02-28
AU2007286808A1 (en) 2008-02-28
JP5448818B2 (en) 2014-03-19
AR062468A1 (en) 2008-11-12
TWI411614B (en) 2013-10-11
PE20080611A1 (en) 2008-07-21
EP2069354A1 (en) 2009-06-17
MX2009001878A (en) 2009-03-03

Similar Documents

Publication Publication Date Title
EP2069354B1 (en) Aza-benzofuranyl compounds and methods of use
EP2222675B1 (en) 5-anilinoimidazopyridines and methods of use
AU2007286807B2 (en) Aza-benzothiophenyl compounds and methods of use
AU2008343062B2 (en) 8-Anilinoimidazopyridines and their use as anti-cancer and/or anti-inflammatory agents
EP2099796B1 (en) Aza-indolyl compounds and methods of use
WO2010003025A1 (en) Bicyclic heterocycles as mek kinase inhibitors
RU2448111C2 (en) Azabenzofuranyl compounds and methods of use

Legal Events

Date Code Title Description
WWE Wipo information: entry into national phase

Ref document number: 200780039152.0

Country of ref document: CN

121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 07814272

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 2007286808

Country of ref document: AU

WWE Wipo information: entry into national phase

Ref document number: 867/DELNP/2009

Country of ref document: IN

WWE Wipo information: entry into national phase

Ref document number: 196956

Country of ref document: IL

Ref document number: 12009500274

Country of ref document: PH

WWE Wipo information: entry into national phase

Ref document number: 2660546

Country of ref document: CA

WWE Wipo information: entry into national phase

Ref document number: MX/A/2009/001878

Country of ref document: MX

WWE Wipo information: entry into national phase

Ref document number: 2009525720

Country of ref document: JP

WWE Wipo information: entry into national phase

Ref document number: 2007814272

Country of ref document: EP

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 2007286808

Country of ref document: AU

Date of ref document: 20070820

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: 1020097005786

Country of ref document: KR

ENP Entry into the national phase

Ref document number: 2009110144

Country of ref document: RU

Kind code of ref document: A

ENP Entry into the national phase

Ref document number: PI0714635

Country of ref document: BR

Kind code of ref document: A2

Effective date: 20090225