CA2238310A1 - Substituted aryl or heteroarylamides having retinoid-like biological activity - Google Patents

Abstract

Compounds of formula (1), wherein X is CH or N; R1 is independently H or alkyl of 1 to 6 carbons; m is an integer having the value of 0-5; p is an integer having the value of 0-2; r is an integer having the value 0-2; L is -(C=Z)-NH-or -NH-(C=Z)- where Z is O or S; Y is a phenyl or naphthyl group, or heteroaryl selected from a group consisting of pyridyl, thienyl, furyl, pyridazinyl, pyrimidinyl, pyrazinyl, thiazolyl, oxazolyl, imidazolyl and pyrrazolyl, said phenyl, naphthyl and heteroaryl groups being optionally substituted with one or two R1 groups; A is (CH2)q where q is 0-5, lower branched chain alkyl having 3-6 carbons, cycloalkyl having 3-6 carbons, alkenyl having 2-6 carbons and 1 or 2 double bonds, alkynyl having 2-6 carbons and 1 or 2 triple bonds, and B is COOH or a pharmaceutically acceptable salt thereof, COOR8, CONR9R10, -CH2OH, CH2OR11, CH2OCOR11, CHO, CH(OR12)2, CHOR13O, -COR7, CR7(OR12)2, CR7OR13O, where the remaining substituents are defined in the claims, have retinoid-like biological activity.

Description

CA 022383l0 l998-0~-2l W O 97/19062 PCT~US96/18529 SUBSTI~ ~ ARYL OR ~ ~O~T-~IDES HAVING
2RETINOID-LIKE BIOLOGICAL ACTlv~Y
3BACRGROUND OF THE ~ v~:N-lON
4 1. Field of the Invention ~The present invention re~ates to novel compounds 6having retinoid-like biological activity. More 7specifically, the present invention relates to 8amides formed between aryl or heteraryl amines and ~ aryl or heteroaryl carboxylic acids where one of the aromatic or heteroaromatic moieties bears an electron withdrawing substituent. The compounds 12 have retinoid-like biological activity.
13 2. Background Art 4 Compounds which have retinoid-like activity are well known in the art, and are described in numerous 6 United States and other patents and in scientific 17 publications. It is generally known and accepted in 18 the art that retinoid-like activity is useful for 19 treating ~nim~ls of the m~mm~lian species, including hl-m~ns, for curing or alleviating the symptoms and 21 conditions of numerous diseases and conditions. In 22 other words, it is generally accepted in the art 23 that pharmaceutical compositions having a 24 retinoid-like compound or compounds as the active 26 ingredient are useful as regulators of cell 26 proliferation and differentiation, and particularly 27 as agents for treating skin-related diseases, 28 including, actinic keratoses, arsenic keratoses, inflammatory and non-inflammatory acne, psoriasis, ichthyoses and other keratinization and 31 hyperproliferative disorders of the skin, eczema, 32 atopic dermatitis, Darriers disease, lichen planus, 33 prevention and reversal of glucocorticoid damage CA 02238310 1998-0~-21 W O 97/19062 PCTnUS96/18529 1 (steroid atrophy), as a topical anti-microbial, as 2 skin anti-pigmentation agents and to treat and 3 reverse the e~fects of age and photo damage to the 4 skin. Retinoid compounds are also useful for the 5 prevention and treatment of cancerous and 6 precancerous conditions, including, premalignant and 7 malignant hyperproli~rative diseases such as 8 cancers of the breast, ~kin, prostate, cervix, 9 uterus, colon, bladder, esophagus, stomach, lung, larynx, oral cavity, ~lood and lymphatic system, t1 metaplasias, dysplasias, neoplasias, leukoplakias 12 and papillomas of the mucous membranes and in the 13 treatment o~ Kaposi's sarcoma. In addition, 14 retinoid compounds can be used as agents to treat 15 diseases of the eye, including, without limitation, 16 proliferative vitreoretinopathy (PVR), retinal 17 detachment, dry eye and other corneopathies, as well 18 as in the treatment and prevention of various 19 cardiovascular diseases, including, without 20 limitation, diseases associated with lipid 21 metabolism such as dyslipidemias, prevention of 22 post-angioplasty restenosis and as an agent to 23 increase the level of circulating tissue plasminogen 24 activator (TPA). Other uses for retinoid compounds 25 include the prevention and treatment of conditions 26 and diseases associated with human papilloma virus 27 (HPV), including warts and genital warts, various 28 inflammatory diseases such as pulmonary fibrosis, 2~ ileitis, colitis and K:rohn's disease, 30 neurodegenerative dise,ases such as Alzheimer's 31 disease, Parkinson's d.isease and stroke, improper 32 pituitary function, in~luding insufficient 33 production of growth hormone, modulation of CA 02238310 1998-0~-21 W O 97/19062 PCT~US96/18529 apoptosis, including both the induction of apoptosis and i nh i hition of T-Cell activated apoptosis, 3 restoration of hair growth, including combination therapies with the present compounds and other 5 agents such as MinoxidilR, diseases associated with 6 the immllne system, including use of the present 7 compounds as immllnosuppressants and 8 ;mmllnostimulants, modulation of organ transplant o rejection and facilitation of wound healing, including modulation of chelosis.
United States Patent No. 4,723,028 (Shudo), Published European Patent Application Nos. 0 170 105 (Shudo), German Patent Application No. DE 3524199 Al ~Shudo), PCT WO 91/16051 (Spada et al.), PCT WO
~ 85/04652 (Polus) and J. Med Chem. 1988 31, 2182 -16 2192 (Kagechika et al.), describe or relate to aryl and heteroary or diary substituted olephines or amides having retinoid-like or related biological 19 activity.
United States Patent Nos. 4,992,468, 5,013,744, 21 5,068,252, 5,175,185, 5,202,471, 5,264,456, 22 5,324,840, 5,326,898, 5,349,105, 5,391,753, 23 5,414,007 and 5,434,173 ~assigned to the same 24 assignee as the present application) and patents and 2s publications cited therein, describe or relate to 26 compounds which have retinoid-like biological n activity and a structure wherein a phenyl and a 28 heteroaryl or a phenyl and a second phenyl group is 2~ linked with an olephinic or acetylenic linkage.
Still further, several co-pending applications and 31 recently issued patents which are assigned to the 32 assignee of the present application, are directed to 33 further compounds having retinoid-like activity.

CA 022383l0 l998-05-2l W O 97/19062 PCTAJS96tl8529 _, 1 It is now general knowledge in the art that two 2 main types of retinoid receptors exist in m~m~ s 3 ( and other organisms). The two main types or 4 families of receptors are respectively designated RARS and RXRS . Within each type there are subtypes;
~ in the RAR ~amily the subtypes are designated RARar 7 RA~B and RARr, in RXR the subtypes are: RXRa , RX~3B and 8 ~XRr. It has also been esta}:)lished in the art that ~ the distribution of the two main retinoid receptor types, and of the several sub-types is not uniform in the various tissues and organs of mAmm~lian 12 organisms. Accordingly, among compounds capable of 13 blnding to retinoid receptors, speci:Eicity or 14 selectivity :Eor one o~ the main types or families, ts and even speci:ficity or selectivity :Eor one or more 16 subtypes within a family o~ receptors, is considered a desirable pharmacological property.
he present invention provides compounds having 19 retinoid-like biological activity and speci~ically 20 compounds which bind to one or more RAR retinoid 21 receptor subtypes.
22 SUMMARY OF ~EIE Irlv~ ION
23 The present invention covers compounds oi~
24 Formula 1 26 (R1)m 22 ~ X \
~P

33 For~nula 1 CA 022383l0 l998-05-2l W O 97/l9062 PCT~US96/18S29 1 wherein X is CH or N;
2 Rl is independently H or alkyl of 1 to 6 3 carbons;
4 m is an integer having the value of 0 -- 5;
p is an integer having the value of 0 -- 2; r 6 iS an integer having the value O - 2;
7 L is -~C=Z)-NH- or -NH-~C=Z)- where Z is O or S;
8 Y is a phenyl or naphthyl group, or heteroaryl 9 selected from a group consisting of pyridyl, o thienyl, furyl, pyridazinyl, pyrimidinyl, pyrazinyl, thiazolyl, oxazolyl, imidazolyl and pyrrazolyl, said 2 phenyl, naphthyl and heteroaryl groups being 3 optionally substituted with one or two Rl groups;
4 W is a su~stituent selected from the group consisting of F, Br, Cl, I, Cl_6alkyl, fluoro 6 substituted Cl_6 alkyl, NO2, N3, OH, OCH20CH3, 7 OCl_l0alkyl, tetrazol, CN, SO2Cl_6--alkyl,SO2C1_6--alkyl, 8 SO2C1_6-fluoro substituted alkyl, SO-Cl_6 alkyl, 1~ CO-Cl6alkyl, COOR8, phenyl, phenyl itself substituted with a W group other than with phenyl or substituted 21 phenyl with the proviso that when X is CH and r is 0 ~ then p is not 0 and at least one W group is not 23 alkyl;
24 A is ~CH2)q where q is 0-5, lower branched chain 25 alkyl having 3-6 carbons, cycloalkyl having 3-6 26 carbons, alkenyl having 2-6 carbons and 1 or 2 27 double bonds, alkynyl having 2--6carbons and 1 or 2 28 triple bonds, and 2~ B is COOH or a pharmaceutically acceptable salt 30 thereof, COOR8, CONR9R1o, -CH20H, CH20Rll, CH20CORll, 31 CHO, CH(ORl2)2, CHORl3O, -COR7~ CR7(ORl2)2~ CR7ORl3O, 32 where R7 is an alkyl, cycloalkyl or alkenyl group 33 containing 1 to 5 carbons, R8 is an alkyl group of CA 022383l0 l998-0~-2l W O 97/19062 PCT~US96tl8529 t to 10 carbons or trimethylsilylalkyl where the alkyl 2 group has 1 to 10 aarbons, or a cycloalkyl group of 3 5 to 10 carbons r or R8 is phenyl or lower 4 alkylphenyl, R9 and R~o independently are hydrogen, ~n alkyl group of 1 to 10 carbons, or a cycloalkyl 6 group of 5-10 carbons, or phenyl or lower 7 alkylphenyl, Rl1 is lower alkyl, phenyl or lower 8 alkylphenyl, Rl2 is lower alkyl, and R13 is divalent ~ alkyl radical of 2-5 carbons.
In a second aspect, this invention relates to 11 the use of the compounds of Formula 1 for the 12 treatment of skin-related diseases, including, 13 without limitation, actinic keratoses, arsenic 14 keratoses, inflammatoxy and non--inflammatory acne, 16 psoriasis, ichthyoses and other keratinization and 16 hyperproliferative di;orders of the skin, eczema, 7 atopic dermatitis, Darriers disease, lichen planus, 18 prevention and reversal of glucocorticoid damage 19 (steroid atrophy~, as a topical anti-microbial, as skin anti-pigmentation agents and to treat and 21 reverse the effects of age and photo damage to the 22 skin. The compounds are also useful for the 23 prevention and treatment of cancerous and 24 precancerous conditions, inc~uding, premalignant and 26 malignant hyperproliferative diseases such as 26 cancers of the breast~ skin, prostate, cervix, 27 uterus, colon, bladder, esophagus, stomach, lung, 28 larynx, oral cavity, blood and lymphatic system, 2~ metaplasias, dysplasias, neoplasias, leukoplakias and papillomas of the mucous membranes and in the 31 treatment of Kaposi's sarcoma. In addition, the ~ present compounds can be used as agents to treat 33 diseases of the eye, including, without limitation, CA 02238310 1998-0~-21 W O 97/19062 PCT~US96/18529 1 pro~iferative vitreoretinopathy (PVR), retinal 2 detachment, dry eye and other corneopathies, as well 3 as in the treatment and prevention of various 4 cardiovascular diseases, including, without limitation, diseases associated with lipid 6 metabolism such as dyslipidemias, prevention of 7 post-angioplasty restenosis and as an agent to 8 increase the level of circulating tissue plasminogen 9 activator (TPA). Other uses for the compounds of o the present invention include the prevention and 11 treatment of conditions and diseases associated with 12 human papilloma virus (HPV), including warts and 3 genital warts, various inflammatory diseases such as 4 pulmonary fibrosis, ileitis, colitis and Krohn's disease, neurodegenerative diseases such as 6 Alzheimer's disease, Parkinson's disease and stroke, 7 improper pituitary ~unction, including insufficient 8 production of growth hormone, modulation of 19 apoptosis, including both the induction of apoptosis 20 and ; nh i hition of T-Cell activated apoptosis, 21 restoration of hair growth, including combination 22 therapies with the present compounds and other 23 agents such as MinoxidilR, diseases associated with 24 the ;mmllne system, including use of the present 25 compounds as ;mmtlnosuppressants and 26 i mmtlno5timulants ~ modulation of organ transplant 27 rejection and facilitation of wound healing, 28 including modulation of chelosis.
2~ This invention also relates to a pharmaceutical 30 formulation comprising a compound of Fon~ula 1 in 31 admixture with a pharmaceutically acceptable 32 excipient.
33 In another aspect, this invention relates to WO 97/19062 PCT~US96/18529 1 processes for making a compound of Formula 1 which 2 processes comprise reacting, in the presence of an 3 acid acceptor or water acceptor, a compound o~
4 Formula 2 with a compound of Formula 3 where X1 is ~ OH, halogen, or other group which renders the -COX
6 group reactive ~or amide ~ormation, and where the 7 remaining symbols are defined as in connection with 8 Formula 1. Alternatively, the process o~ the g invention comprises re~cting a compound of Formula 2a with a compound of Formula 3a, where the symbols 11 are defined as above.

14 m(Rl) \~
16 ~ COXlH2N-'Y(W~r-A-B
17 ~W~p~

22 Formula 2 Formula 3 m(Rl 26 \~
27 ~p ~ NH2 XIOC-Y(W)r-A-B

33 Formula 2a For~nula 3a CA 022383l0 l998-0~-2l W O 97/19062 PC~US96/18529 1 Still further, the present invention relates to 2 such reactions performed on the compounds of Formula 3 1 which cause transformations of the B group while 4 the reaction product still remains within the scope of Formula 1.
6 General Em~o~; - tsDefinitions 7 The term alkyl refers to and covers any and all 8 groups which are known as normal alkyl, g branched-chain alkyl and cycloalkyl. The term alkenyl refers to and covers normal alkenyl, branch 11 chain alkenyl and cycloalkenyl groups having one or 12 more sites oE unsaturation. Similarly, the term 13 alkynyl refers to and covers normal alkynyl, and 14 branch chain alkynyl groups having one or more 15 triple bonds.
16 Lower alkyl means the a~ove-defined broad 17 definition of alkyl groups having 1 to 6 carbons in 18 case o~ normal lower alkyl, and as applicable 3 to 6 1~ carbons for lower branch chained and cyc~oalkyl 20 groups. Lower alkenyl is de~ined similarly having 2 21 to 6 carbons for normal lower al~cenyl groups, and 3 22 to 6 carbons for branch chained and cyclo- lower 23 alkenyl groups. Lower alkynyl is also defined 24 similarly, having 2 to 6 carbons for normal lower 25 alkynyl groups, and 4 to 6 carbons for branch 26 chained lower alkynyl groups.
27 The term "ester" as used here refers to and 28 covers any compound falling within the definition of 29 that term as classically used in organic chemistry.
30 It includes organic and inorganic esters. Where B
31 of Formula 1 is --COOH,this term covers the products 32 derived from treatment of this function with 33 alcohols or thioalcohols pref~erably with aliphatic CA 02238310 1998-0~-21 1 alcohols having 1-6 carbons. Where the ester is 2 derived from compounds where B is --CH20~I, this term 3 covers compounds derived from organic acids capable 4 of forming esters including phosphorous based and sulfur based acids, or compounds of the formula 6 -CH20CORll where Rll is any substituted or 7 unsubstituted aliphatic, aromatic, heteroaromatic or 8 aliphatic aromatic group, preferably with 1-6 g carbons in the aliphatic portions.
o Unless stated otherwise in this application, preferred esters are derived from the saturated 12 aliphatic alcohols or acids of ten or fewer carbon 13 atoms or the cyclic or saturated aliphatic cyclic 14 alcohols and acids of 5 to 10 carbon atoms.
1~ Particularly pre~erred aliphatic esters are those 16 derived Irom lower alk~yl acids and alcohols. Also 17 preferred are the phen~yl or lower alkyl phenyl 8 esters.
19 Amides has the meaning classically accorded that 20 term in organic chemisl_ry. In this instance it 21 includes the unsubstituted amides and all aliphatic 22 and aromatic mono- and di- substituted amides.
~ Unless stated otherwise in this application, 24 preferred amides are the mono- and di--substituted amides derived from the saturated aliphatic radicals 26 of ten or fewer carbon atoms or the cyclic or 27 saturated aliphatic-cyclic radicals of 5 to 10 28 carl~on atoms. Particu],arly preferred amides are 29 those derived ~rom substituted and unsubstituted lower alkyl amines. A]so preferred are mono- and 31 disubstituted amides derived from the substituted 32 and unsubstituted phenyl or lower alkylphenyl 33 amines. Unsubstituted amides are also preferred.

CA 02238310 1998-0~-21 W O 97/19062 PCT~US96/18529 1 Acetals and ketals include the radicals of the 2 formula-CK where K is (-OR) 2~ Here, R is lower 3 alkyl. Al~o, K may be --OR70-- where R~ is lower alkyl 4 of 2-~ carbon atoms, straight chain or branched.
A pharmaceutically acceptable salt may be 6 prepared for any compounds in this invention having 7 a functionality capable of forming such-salt, for 8 ~xAmrle an acid functiona~ity. A p~armaceutically 9 acceptable salt is any salt which retains the activity of the parent compound and does not impart 11 any deleterious or untoward ef~ect on the subject to t2 which it is a~m; n; stered and in the context in which 13 it is a~m; n; stered. Pharmaceutically acceptable 14 salts may be derived from organic or inorganic 16 bases. The salt may be a mono or polyvalent ion.
16 Of particular interest are the inorganic ions, 17 sodium, potassium, calcium, and magnesium. Organic 8 salts may by be made with amines, particularly 19 ammonium salts such as mono-, di- and trialkyl amines or ethanol amines. Salts may also be formed 21 with caffeine, tromethamine and similar molecules.
22 Where there is a nitrogen sufficiently basic as to 23 be capable of forming acid addition salts, such may 24 be formed with any inorganic or organic acids or alkylating agent such as methyl iodide. PreEerred 26 salts are those formed with inorganic acids such as 27 hydrochloric acid, sulfuric acid or phosphoric acid.
28 Any of a number of simple organic acids such as 29 mono-, di-- or tri-- acid may also be used.
~ Some of the compounds of the present invention 31 may have trans and cis (E and Z) isomers. In 32 addition, the compounds of the present invention may 33 contain one or more chiral centers and therefore may W O 97/19062 PCT~US96/18529 1 exist in enantiomeric ~nd diastereomeric forms. The 2 scope of the present :invention is intended to cover 3 all such isomers per se, as well as mixtures of cis 4 and trans isomers, mixtures of diastereomers and 6 racemic mixtures of enantiomers (optical isomers) as 6 well.
7 With reEerence to the symbol X in Formula 1, 8 compounds are equally preferred where X is CH or N.
9 When X i5 CH then the benzene ring is pre~erably 1, o 3, 5 substituted with the L group occupying the 1 11 position and the W anfl/or Rl groups occupying the 3 12 and 5 positions. When the symbol X is N, then the t3 pyridine ring is preferably 2,4,6 substituted with 14 the L group occupying the 4 position ~nd the W
~5 and/or Rl groups occupying the 2 and 6 positions.
16 The 1:. group o:E Fonnula 1 is preferably 17 -(C=Z)-NH-, and Z is preferably 0. In other words, 18 those carbamoyl or amide compounds are pre:Eerred in 19 accordance with the present invention where the -NH-moiety is attached, to the Y group.
21 Referring now to the W group in Formula 1, this 22 group is, generally speaking~ an electron 23 withdrawing group. W is present in the aompounds of 24 the invention either in the phenyl or pyridyl ring 2~ (shown in Formula 1 as substituent "~W)p") 2nd/or as 2~ a substituent oE the aryl or heteroaryl group Y.
27 Pre~erably, the W grou,p is present in the Y group, 28 or both in the Y group and in the phenyl or pyridyl 25~ ring discussed above. In the aryl or heteroaryl moiety the W group is preferably located in the 31 position adjacent to the A-B group; pref~erably the 32 A-B group is in Para position in the phenyl ring 33 relative to the L (amide or carbamoyl) moiety, and CA 022383l0 l998-0~-2l W O 97/19062 PCT~US96/18529 1 therefore the W group is preferably in meta position 2 relative to the L (amide or carbamoyl) moiety.
3 Preferred W groups are F, NO2, Br, I, CF3, N3, and - 4 OH. Alternatively, in the phenyl or pyridyl ring (shown in Formula 1 as substituent "(W)y") W is an 6 alkyl group, preferably branch-chained alkyl, such 7 as tertiary butyl, and preferably p is 2. Moreover, 8 the presence of one or two fluoro substituents in g the Y group is especially preferred. When the Y
0 group is phenyl, the Eluoro substituents preferably 11 are in the ortho and ortho' positions relative to 12 the A--B group.
13 With reference to the symbol Y in Formula 1, the 14 preferred compounds o~ the invention are those where Y is phenyl, pyridyl, 2-thiazolyl, thienyl, or 16 furyl, more preferably phenyl. As far as 17 substitutions on the Y (phenyl) and Y (pyridyl) 18 groups are concerned, compounds are preferred where 19 the phenyl group is 1,4 (~ara) substituted by the and A-B groups, and where the pyridine ring is 2,5 21 substituted by the L and A--B groups. (Substitution 22 in the 2,5 positions in the "pyridine" nomenclature 23 corresponds to substitution in the 6-position in the 24 "nicotinic acid" nomenclature.) In the preferred compounds of the invention there is no optional R
26 substituent (other than H) on the Y group.
27 The Rl groups, when present, preferably are H or 28 CH3.
29 The A-B group of the preferred compounds is (CH2)n-COOH or (CH2)n-COOR8, where n and R8 are defined 31 as above. Even more pre~erably n is zero and R8 is 32 lower alkyl, or n is zero and B is COOH or a 33 pharmaceutically acceptable 5 alt thereof.

W O 97/19062 PCT~US96/18529 1 The most preferred compounds of the invention 2 are shown in ~able :L, with reference to Formula 4.

~W~

Formula 4 Table 1 Compound # X Wl W2 W3 R 8 1 N H F H Et 3 N H H H Et 22 5 CH H F H Et 24 7 CH OH F H Et 26 9 N H F F Me 28 11 CH H F F Me 2~ 12 CH H F F H
13 N H NO2 H Me ~ 15l CH H H H H
33 lCompound 15 is prior art, described in J. Med Chem.
34 1988, 31, 2182 (Kaqechika et al.) CA 02238310 1998-0~-21 W O 97/19062 PCT~U~96/18529 ~odes of Z~ministration 2 The compounds of this invention may be 3 a~m; ni stered systemically or topically, depending on 4 such considerations as the condition to be treated, need for site-specific treatment, quantity of drug 6 to be a~m; n; stered, and numerous other 7 considerations.
8 In the treatment of ~ermatoses, it will g generally be preferred to a~m; n; ster the drug topically, though in certain cases such as treatment 11 of severe cystic acne or psoriasis, oral 12 a~m;n; stration may also be used. Any common topical 13 formulation such as a solution, suspension, gel, 14 ointment, or salve and the like may be used.
15 Preparation of such topical formulations are well 16 described in the art o~ pharmaceutical formulations 17 as exemplified, for example, Remington's 18 Pharmaceutical Science, Edition 17, Mack Publishing 1~ Company, Easton, Pennsylvania. For topical 20 application, these compounds could also be 21 a~m; n; stered as a powder or spray, particularly in 22 aerosol form. If the drug is to be administered 23 systemically, it may be confected as a powder, pill, 24 tablet or the like or as a syrup or elixir suitable for oral a~lm; n; stration. For intravenous or 26 intraperitoneal administration, the compound will be 27 prepared as a solution or suspension capable of 28 being a~ministered by injection. In certain cases, 29 it may be useful to formulate these compounds by 30 injection. In certain cases, it may be useful to 31 formulate these compounds in suppository form or as 32 extended release formulation for deposit under the 33 skin or intramuscular injection.
34 Other medicaments can be added to such topical 36 formulation for such secondary purposes as treating 36 skin dryness; providing protection against light;

CA 022383l0 l998-0~-2l 1 other medications for treating dermatoses;
2 medicaments for preventing infection, reducing 3 irritation, in:~lammat:ion and the like.
4 Treatment of derm.atoses or any other indications 5 known or discovered to be susceptible to treatment 6 l~y retinoic acid-like compounds will be effected by 7 a~m;n;stration of the therapeutically efi~ective dose 8 of one or more compounds of the instant invention.
~ A therapeutic concentration will be that concentration which e~fects reduction o~ the 11 particular condition, or retards it expansion. In 12 certain instances, the compound potentially may be used in prophylactic manner to prevent onset o:E a 14 particular condition.
A usef~ul therapeutic or prophylactic 16 concentration will vary from condition to condition and in certain instance~ may vary with the severity 18 of the condition being treated and the patient's 19 susceptibility to trea.tment. Accordingly, no single 20 concentration will be uniformly usef~ll, but will 21 require modification d.epending on the 22 particularities oi~ the disease being treated. Such 23 concentrations can be arrived at through routine 24 experimentation. However, it is anticipated that in 25 the treatment of, for example, acne, or similar 26 dermatoses, that a formulation containing between 27 0 . 01 and 1.0 milligrams per mililiter of formulation 28 Will constitute a therapeutically effective 2~ concentration for total application. If 30 a-lm; n ~stered systemically, an amount between O.01 31 and 5 mg per kg per day of body weight would be 32 expected to effect a t.herapeutic result in the 33 treatment of many disease for which these compounds 34 are useful.
Assay of Retinoid--like Bioloqical Activity 36 The retinoid-like activity of the compounds oi~

CA 02238310 1998-0~-21 W O 97/19062 PCTrUS96/185Z9 _ .

the invention can be confirmed in assays wherein ~ 2 ability o~ the compound to bind to retinoid 3 receptors is measured. As it i~ noted in the - 4 introductory section of this application for patent 5 two main types o~ retinoic acid receptors (RAR and 6 RXR) exist in m~mm~ lS ~ and other organisms). Within 7 each type there are sub-types (RARa r RARB ~ RARr ~ RX~
8 RXR~ and RXRr~ the distribution of which is not ~ uniform in the various tissues and organs of 10 m~mm~ lian organisms. Selective binding of only one 11 or two retinoid receptor subtypes within one 12 retinoid receptor family can give rise to beneficial 13 pharmacological properties because of the varying 14 distribution of the sub-types in the several 16 m~mm~ lian tissues or organs. For the 16 above-sllmm~rized reasons, binding of any or all of 17 the retinoid receptors, as well as ~pecific or 18 selective activity in a receptor family, or 19 selective or specific activity in any one o~ the 20 receptor subtypes, are all considered desirable 21 pharmacological properties.
22 In light o~ the ~oregoing the prior art has 23 developed assay procedures for testing the agonist 24 like activity of compounds in the RAR~, RARB ~ RARr 25 RXRa, RXRB and RXRr receptor subtypes. For ~r~mrle~
26 a chimeric receptor transactivation assay which 27 tests for agonist-like activity in the RARa~ RARB~
28 RAR~ , and RXRa receptor subtypes, and which is based 2~ on work published by Feigner P. L. and Holm M.
~ (1989) Focus, 11 2 is described in detail in U.S.
31 Patent No. 5 r 455,265. The specification of United 32 States Patent No. 5~455,265 is expressly 33 incorporated herein by re~erence.
A holoreceptor transactivation assay and a -~5 ligand binding assay which measure the ability of 36 the compounds of the invention to bind to the CA 022383l0 l998-05-2l WO 97/19062 PCT~US96/18529 1 several retinoid receptor subtypes, respectively, 2 are described in pub].ished PCT Application No. WO
3 W093/11755 (particularly on pages 30 - 33 and 37 -4 41) published on June 24, 1993, the speci~ication o~
which is also incorporated herein by re~erence. A
6 description of the ligand binding assay is al~o 7 provided below.
8 BI~DING ASSAY
g All binding assay~ were performed in a similar fashion. All six receptor types were derived from 11 the expressed receptor type (RA~ a, ~, r and RXR a, 12 33, r ) expressed in Baculovirus. Stock solutions of 13 all compounds were prepared as lOmM ethanol 14 solutions and serial dilutions carried out into 1:1 DMSO; ethanol. Assay buffers consisted of the 16 following l~or all six receptor assays: 8g6 glycerol, 17 120mM KCl, 8mM Tris, 5mM CHAPS 4mM DTT and O.24mM
18 PMSF, pH - 7.4@ room temperature.
19 All receptor biding assays were performed in the 20 same manner. The final assay volume was 250,u1 and 21 contained ~rom 10-40~g oE extract protein depending 22 on receptor being assayed along with 5 nM of [3H]
23 all-trans retinoic acid or lOnM [3H] 9-cis retinoic 24 acid and varying concentrations oi~ competing ligand 25 at concentrations that ranged from O - 10 -5 M. The 26 assays were formatted for a 96 well minitube system.
27 Incubations were carried out at 4~C until 28 equilibrium was achieved. Non-specific binding was 29 defined as that binding remaining in the presence of 30 lOOOnM of the appropriate unlabeled retinoic acid 31 isomer. At the end of the incubation period, 50~1 32 of 6.25% hydroxyapitite was added in the appropriate 33 wash buffer. The wash buffer consisted of lOOmM
34 KCl, lOmM Tris and either 5mM CHAPS (RXR a, 13, r) or 35 0. 596 Triton X--100 (RAR a, 13, r ) . The mixture was 36 vortexed and incubated for 10 minutes at 4~C, CA 02238310 1998-0~-21 W O 97/19~62 PCTnJS96/18529 1 centrifuged and the supernatant removed. The 2 hydroxyapitite was washed three more times with the 3 appropriate wash bu~er. The receptor-ligand ~ 4 complex was adsorbed by the hydroxyapitite. The 6 amount of receptor-ligand complex was determined by 6 liquid scintillation counting of hydroxyapitite 7 pellet.
~ After ~orrecting for non-speci~ic bindin~ IC5~
9 values were determined. The IC50 value is dei~ined as the concentration of competing ligand needed to 11 reduce specific binding by 50%. The IC50 value was 12 determined graphically from a loglogit plot of the 13 data. ~he Kd values were determined by application 14 of the Cheng-Prussof equation to the IC50 values, the 15 labeled ligand concentration and the Kd of the 16 labeled ligand. The results of ligand b; n~; ~g 17 assay are expressed in Kd numbers. (See Cheng et al.

18 Biochemical Pharmacology Vol. 22 pp 3099-3108, 19 expressly incorporated herein by reference.~
Table 2 shows the results of the ligand binding 21 assay for certain exemplary compounds o~ the 22 invention.
23 ~RT.~ 2 24 Ligand Binding Assay 25 Compound #Kd (nanomolar) 26 RARaRAR~ Ru~Rr RXRa RXR~ RXRr 27 2 14.00 O.OO O.O0 O.OO O.OO O.OO
28 ~I 19 ~ 00 0 . 00 0 . 00 0 _ 00 0 . 00 0 _ 00 29 6 26.0 0.00 0.00 0.00 0.00 o.oo 8 77.0 0.00 0.00 0.00 0.00 0.00 31 10 62.0 0.00 0.00 0.00 0.00 O.oo 32 12 87.0 0.00 0.00 0.00 O.Oo 0.00 33 14 94.0 0.00 0.00 0.00 0.00 0.00 34 151 37.0 0.00 0.00 0.00 0.00 0.00 35 0 . 00 indicates ~alue greater than lOOOnM
36 ( nanomolar)1~ompound 15 is prior art, described in J.

CA 022383l0 l998-05-2l WO 97/19062 PCT~US96/18529 1 Med Chem. 1988, 31, 2182 (Kagechika et al.) 2 As it can be seen from the test results 3 sllmm~-ized in Table 2, the therein indicated 4 exemplary compounds of the invention bind 5 specifically or selectively to RAR~ receptors.

8 ~Iormones ~ All trans-Retinoic acid (t-RA) (Sigma Chemicals Co., St. Louis, MO) was stored at -70~C. Prior to 11 each experiment the co~mpound was dissolved in 100%
12 ethanol at l mM and diluted in culture medium 13 immediately before use. All experiments were 14 performed in subdued light. Controls were assayed using the same concentration of ethanol as present 16 in the experimental plates and this concentration of 17 diluent had no effect in either assay.
18 Cells and Cell Culture 1~ All cell lines, RPMI 8226, ME-180 and AML-193 20 were obtained from the American Type Culture 21 Collection (ATCC~ Rock~ille, MD~. RPMI 8226 is a 22 human hematopoietic ce:Ll line obtained from the 23 peripheral blood of a patient with multiple myeloma.

24 The cells resemble the lymphoblastoid cells of other 26 human lymphocyte cell lines and secrete a-type light 26 chains of ;mmllnoglobulin. RPMI-8226 cells are grown 27 in RPMI medium (Gibco~ supplemented with 10% fetal 28 bovine serum, glutamine and antibiotics. The cells 2~ were maintained as suspension cultures grown at 37~C
in a humidified atmosphere of 5~ CO2 in air. The 31 cells were diluted to a concentration of l x 105/ml 32 twice a week.
33 ME-180 is a human epidermoid carcinoma cell line 34 derived from the cervix. The tumor was a highly 3s invasi~e squamous cell carcinoma with irregular cell 3~ cluster~,and no significant keratinization. ME-180 CA 022383l0 l998-0~-2l 1 cells were grown and maintained in McCoy's 5a medium 2 (Gibco) supplemented with 1096 fetal bovine serum, 3 glutamine and antibiotics. The cells were - 4 maintained as monolayer cultures grown at 37~C in a 5 humidified atmosphere of 5% C02 in air. The cells 6 were diluted to a concentration of 1 x 105/ml twice a 7 week.
8 A ~ -193 was established from the blast cells 9 classified as M5 Acute Monocyte Leu}~emia. The growth factor, granulocyte colony-stimulation factor (GM-CSF) was required to establish this cell line t2 and growth factors are necessary for its continuous 3 proliferation in chemically defined medium. AML-193 ~4 cells were grown and maintained in Iscove's modified Dulbecco's medium supplemented with 10% fetal bovine 16 serum, glutAmine and antibiotics with 5~g/ml insulin 17 (Sigma Chemical Co.) and 2 ng/ml rh GM-CSF (R and D
8 Systems). The cells were diluted to a concentration 19 of 3 x 1 O5 /ml twice a week.
Incorporation of 3H--Thymidine 21 The method used for determination of the 22 incorporation of radiolabeled thymidine was adapted 23 :Erom the procedure described by Shrivastav et al.
24 RPMI-8226 cells were plated in a 96 well round bottom microtiter plate (Costar) at a density of 26 1, OOO cells/well. To appropriate wells, retinoid 27 test compounds were added at the final 28 concentrations indicated for a final volume of 150 29 ~l/well. The plates were incubated for 96 hours at 37~C in a humidified atmosphere of 5% C02 in air.
31 Subsequently, 1 ~Ci of [5'-3H]-thymidine (Amersham, 32 U.K. 43 Ci/mmol specific activity) in 25 ,ul culture 33 medium was added to each well and the cells were 34 incubated for an additional 6 hours. The cultures were further processed as described below.
36 M~5--180wells, harvested by trypsinization were CA 022383l0 l998-0~-2l W O 97/19062 PCT~US96/18529 1 plated in a 96 well f:Lat bottom microtiter plate 2 (Costar) at a density of 2,000 cells/well. The 3 cultures were treated as described above for RPMI
4 8226 with the following exceptions. After incubation with thymidine the supernatant was 6 carefully removed, and the cells were washed with a 7 0.5 mM solution of th~nidine in phosphate buffered saline. ME180 cells were briefly treated with 50~1 9 of 2.5% trypsin to dislodge the cells from the plate.
11 AMh-193 cells were plated in a 96 well round 12 bottom microtiter plat:e (Costar) at a density of 13 1,000 cells/well. To appropriate wells, retinoid 14 test compounds were added at the final concentrations indicated for a final volume of 150 16 ~l/well. The plates were incubated for 96 hours at 17 37~C in a humidified aitmosphere of ~96 CO2 in air.
18 Subsequently, 1 ~Ci of {5'-3H]-thymidine (Amershamr 1~ U.K., 43 Ci/mmol specific activity) in 25 ~l culture 20 medium was added to each well and the cells were 21 incubated for an additional 6 hours.
22 All cells lines were then processed as follows:
23 the cellular DNA was p~recipitated with 1096 24 trichloroacetic acid anto glass fiber filter mats 25 using a SKATRON multi-well cell harvester (Skatron 26 Instruments, Sterling VA). Radioactivity 27 incorporated into DNA, as a direct measurement of 28 cell growth~ was measured }~y liquid scintillation 2~ counting. The numbers represent the mean 30 disintegrations per minute of incorporated thymidine 31 f~rom triplicate wells + SEM.
32 In the above notecl in vitro cell lines exemplary 33 ~ompound 2 of the invention caused significant 34 decrease in the proliferation of the tumor cell 35 lines (as measured ~y incorporation of radioactive 36 la~eled thymidine) in the 10-ll to 10-6 molar CA 022383l0 l998-0~-2l 1 concentration range of the test compound.
2 SPECIFIC EMBODI~ENTS
3 The compounds of this invention can be made by - 4 the synthetic chemical pathways illustrated here.
The synthetic chemist will readily appreciate that 6 the conditions set out here are specific embodiments 7 which can be generalized to any and all of the 8 ccmpouncls represented by Formula 1.
Generally speaking the process of preparing compounds of the invention involves the formation of an amide by the reaction of a compound of the 2 general Formula 2 with a compound of general Formula 13 3, or by the reaction of a compound of general 14 Formula 2a with a compound of general Formula 3a as these formulas are defined in the Summary section of 16 the present application for patent. Thus, as is 17 noted above, a compound of Formula 2 is an acid or 8 an ~activated form~ of a carboxylic acid attached to 19 a substituted phenyl (in Formula 1 X is CH) or to a substituted pyridyl (in Formula 1 X is N) nucleus.
21 The term "activated form" of the carboxylic acid ~ should be understood in this regard as such 23 derivative of the carboxylic acid which is capable 24 of forming an amide when reacted with a primary amine of Formula 3. In case of the "reverse amides~
26 the activated form of a carboxylic acid is a 27 derivative ( Formula 3a) that is capable of forming 28 an amide when reacted with a primary amine of 2~ Formula 2a. This, generally speaking, means such derivatives of a carboxylic acid which are normally 31 known and used in the art to form amide linkages 32 with an amine. Examples of suitable forms or 33 derivatives for this purpose are acid chlorides, 34 acid bromides, and esters of the carboxylic acid, particularly active esters, where the alcohol moiety 36 of the ester forms a good leaving group. Presently W O 97/19062 PCT~US96/18529 _ .

1 most preferred as reagents in accordance with 2 Formula 2 (or Formula 3a~ are acid chlorides (Xl is 3 Cl). The acid chlorides of For~ula 2 (or of Formula 4 3a) can be prepared by traditional methods from the corresponding esters (X1 is for example ethyl) by 6 hydrolysis and treatment with thionyl chloride 7 (SOCl2). The acid chlc)rides of Formula 2 (or of 8 Formlala 3a) can also be prepared by direct treatment of the carboxylic acid.s with thionyl chloride, where 0 the carboxylic acid, rather than an ester thereof is available co~me~cially or by a known synthetic 2 procedure. The acid chlorides of Formula 2 (or of 3 Formula 3a) are typically reacted with the amine of 14 Formula 3 (or amine of Formula 2a) in an inert solvent, such as methylene chloride, in the presence 16 of an acid acceptor, such as pyridine.
17 The carboxylic acids themselves in accordance 18 with Formula 2 (or Formu~a 3a) are also suitable for 19 amide formation when reacted with an amine, a 20 catalyst (4-dimethylaminopyridine) in the presence 21 o~ a dehydrating agent, such as 22 dicyclohexylcarbodiimide (DCC) or more pere:ferably 23 1--(3-dimethylaminoprop~yl)-3-ethylcarbodiimide 24 hydrochloride (EDC).
The carboxylic aci.ds or the corresponding esters 26 of Formula 2, are generally speaking, prepared as 27 described in the chemical scientific or patent 28 literature and the literature procedures f~or their 29 preparation may be mod:ified, i:~ necessary, by such 30 chemical reactions or processes which per se are 31 known in the art. Reac:tion Scheme 1 provides an 32 example for the preparation of 33 2,6-di-tert-butylisonicotinic acid (Compound C) 34 which is a reagant in accordance with Formula 2 for the preparation of several preferred compounds of 36 the present invention. Thus, .
2~

1 2~6-di-tert-butyl-4-methylpyridine (availa~le 2 cl~mm~l~cially from Aldrich Chemical Co.~ is reacted 3 with N-bromosucc;n;m;de and benzoyl peroxide to ~ 4 provide 4-brr~mnIn~thyl-2,6-di-tert-butylpyridine ~ ( r~ _ und A). Compound A is reacted with ~ase 6 ( sodium hydroxyde) to yield the coresponding 7 hydroxymethyl compound (Co~round B), which i~
8 thereafter oxidized in a Jones oxydation reaction to g give 2,6-d~-tert-butylisonicotinic acid (Compound 10 C).

14 ~er OH
~6 1 ~ ~
16 ~ NBS. ~ZO)7 ~ ~ NaOH
t7 I Ca4 I 1,1 ~o~me 18 eut~N ~tBu Sut ~N ~tBu eu-~ ~N tSu lG
C~mro~m~l A C~ , ' B

23 fO2H

224 Janc'slacctone, ~
26 ~ut N t~u 28 C. , ~ C

~0 36 Reaction Scheme 1 W O 97/19062 PCTfUS96/18529 ~ul~ Br.lHOAc ~ Bu4NBr ~r 6~ ~ },.uJ,~amme 8OE3U ~RU ~6U

~' , D C~ . 'E

t3E~ut ~CO2H
14 tB~i/CO~

1~ ~u 18 C- . 'F

21Reaction Scheme 1 (continued) 22A ~urther exAmple of a compound which serves as 23 a reagent ~or preparing the carbamoyl (or amide) 24 compounds of the present invention 1s pro~ided in 25 ~eaction Sc~e~e 1. 2~4-Di-tert-butylphenol 26 (Aldrich) is bromi~ted in glacial scetic acid to 27 yield 2-bromo-4,6-di-tert-butylphenol (Co~pound D) 28 which is therea~ter reacted with methoxymethyl 2~ chloride (MOMCl)to give O-methoxymethyl-2-~romo-4,6-di-tert-butylphenol 31 ( Compound E). ~ompouna E is treated with t-butyl 32 lithium folowed by carbon dioxide to yield 33 O-methoxymethyl-3,5-di--tert-~utylsalicylic acid ~ (~ompound F~ Compo~na F is a reagent which dif~ers 35 from the compounds gene~ally encompassed ~y Formula 2 only in that the hydroxyl funtion of this compound , W O 97/19062 PCTrUS96/18S29 1 is protected by the methoxymethyl (MOM) group.
2 However, the methoxymethyl protecting group is 3 removed after formation of the carbamoyl (amide) 4 lin}sage, as exemplif~ied in Reaction Scheme 5.
~ ~eaction of an aromatic bromo compound (such as 6 Compound D) with t--butyl lithium followed by carbon 7 dioxide is a preferred method for preparing several 8 aromatic car~oxylic acids in accordance with Formula 2 and Formula 3a, described in the present o application.

12 ~ Na~Cr2O7~ ~OAc. H,SO.~. 9û~C ~ J~co2c2Hs 14 NOz ~) EtO~I/Py. CH2CI. H2N F
16 Compound G
~7 19 T TO2Mf~ TO2Me F~F 1) SOCl_ F~ H71Pd/C ~j/~/r 21 ~ 3) NaN3/H,.O/CH3CN

24 CompoundH (~(~mpolm~

28 ~ C02H 1) SOC17 /~/C0 li 1 ' 7)Mctb~nol/TEA ll l H2N~ N~2H2N~--N~2 CompoundK

36 Reac~ion Scheme 2 W O 97/19062 PCT~US96/18529 1 Reaction Scheme a provides examples for the 2 preparation o:E aromat:ic amino carboxylic acids or 3 esters which serve as reagents corre:3ponding to 4 Formula 3 described above. Thus, in accordance with Reaction Scheme 2, 3-nitro-6-methyl-fluorobenzene 6 (Aldrich) is subjected to oxidation, conversion of 7 the resulting carboxylic acid to an acid chloride 8 and thereai~ter to an ethyl ester ! followed by 9 reduction of the nitro group, to yield ethyl o 2-~luoro-4-amino-benzoate (Compound G). As another 11 example, 2,4,6-trifluorobenzoic acid (Aldrich~ is 12 converted to the meth~l ester through the acid 13 chloride, and the 4-~luoro atom is displaced by 14 reaction with sodium azide to give the intermediate azido compound (Compound }I). Compound H is reduced 16 by hydrogenation, to yield methyl 17 2,6-difluoro-4--aminobenzoate (Compound I). As 18 still another example, 2-nitro-4--aminobenzoicacid 19 (Research Plus Inc.3 i.s converted to its methyl ester (C~ompound K) through the corresponding acid 21 chloride.

CA 022383l0 l998-05-2l (R1)m ~ (R,)m 8 ~ X J a~tone (V~p - ~ J_ CON3 9 FoDm~a2 Fonmula~
t1 tBuOH~eat 13 - _ 14 (R1)m (W)p--~ ~9_ N=C=O

X ~
18 -Fonm~a6 23 (R,)m 28 Fonn~a2a 36 Reaction Scheme 3 W O 97/19062 PCT~US96/18529 =

~ Reaction Scheme 3 illustrates the synthesis of 2 the primary amine compounds oi~ Formula 2a :~rom the 3 acid chlorides (Xl = Cl) or other form of activated 4 acids of Formula 2 where the primary amine o:f~
Formula 2a is not available ~y a pu~lished 6 literature procedure. Thus, substantially in 7 accordance with the steps of a Cllrtius rearrangement, the acid chloride of Formula 2 is reacted with sodium azide in acetone to yield the o azide compound of Formula 5. The azide of Formula 5 is heated in a polar high boiling solvent, such as t-butanol, to provide the intermediate isocyanate of Formula 6, which is hyclrolyzed to yield a compound 14 of Form-lla 2a.

16 F ~ CO2H F ~ CO2Et l.EtO~I2SO4 l 18 ~ \ F 2.BuL~cO2 ~ \ F

21 Suvaw ~aet al Compound L
22 Kogyo Kaguku Zass~
23 197073.97~-979 22 F ~ CO2H F ~ CO2Et 28 ll ¦ l.EtO~n~2SO4 ll l 29 Br ~ 2.BuL~cO2 HO2C ~ F

~euman et~l 33 J. Med Chem- Compound M
34 1995.~,2~31-2540 36 Reaction S~heme 4 CA 022383l0 l998-0~-2l W O 97/19062 PCTrUS96/18529 _.~

1 Reac~ion Scheme 4 illustrates examples for 2 preparing compounds of Formula 3a where such 3 compounds are not available comm~rcially or by a 4 published literature procedure. Thus, by way of 6 example 2,5-difluoro-4-bromobenzoic acid (available 6 by the literature procedure of Sugawara et al. Kogyo 7 Kaguku ~asshi 1970, 73, 972-979, incorporated herein 8 by relference~ is ~irst esterified by treatment with ethyl alcohol and acid to yield the corresponding 0 ester, and thereafter is reacted with butyl lithium 11 followed by carbon dioxide to give the monoester o~
12 2,5-di~luoro terephthalic acid (Compound L). A
3 similar sequence of reactions performed on 4 2,3,5,6-difluoro-4-bromobenzoic acid (available by 15 the literature procedure of Reuman et al. J. Med.
16 Chem. 1995, 38, 2531-2540, incorporated herein by 7 reference) yields the monoester of 8 2,3,5,6-tetrafluoroterephthalic acid ~Compound M) 1~ The just illustrated sequence of reactions can be, 20 generally speaking, utilized for the synthesis of 21 all compounds of Formula 3a with such modi~ication 22 which will become readily apparent to those skilled 23 in the art, where such compounds are not available 24 by a known literature procedure.
Numerous other reactions suitable for preparing 26 compounds of the invention, and for converting 27 compounds of Formula 1 within the scope of the 28 present invention into still further compounds of 2~ the invention, and also for preparing the reagents 30 of Formula 2, Formula 3, Formula 2a and Formula 3a 31 will become readily apparent to those 5killed in the 32 art in light of the present disclosure. In this 33 regard the following general synthetic methodology, 34 applicable for conversion of the compounds of 35 Formula 1 into further homologs and/or derivatives, 36 and also for preparing the reagents of Formula 2 and CA 022383l0 l998-0~-2l WO 97/19062 PCT~US96/18529 1 3, (as well as 2a and 3a) is noted.
2 Carboxylic acids are typically esterified by 3 refluxing the acid in a solution of the appropriate 4 alcohol in the presence of an acid catalyst such as hydrogen chloride or thionyl chloride.
6 Alternatively, the carboxylic acid can be condensed 7 with the appropriate a]Lcohol in the presence of 8 dicyclohexylcarbodiimide and dimethylaminopyridine.
9 The ester is recovered and purified by conventional means. Acetals and ketals are readily made by the 11 method described in March, "Advanced Organic 12 Chemistry," 2nd Edition, McGraw--Hill Book Company, p 13 810). Alcohols r aldehydes and ketones all may be 14 protected by forming respectively, ethers and esters, acetals or ketaLls by known methods such as 16 those described in McOmie, Plenum Publishing Press, 17 1973 and Protecting Groups, Ed. Greene, John Wiley &
18 Sons, 1981.
19 A means for making compounds where A is (CH2)~
(q iS 1 -- 5) iS to subject the compounds of Formula 21 l, where B is an acid or other function, to 22 homologation, using the well known Arndt-Eistert 23 method of homologation, or other known homologation 24 procedures. Similar homologations (and several of the other herein mentioned synthetic 26 transformations) can be transformed on the reagents 27 of Formula 3 or 3a. Compounds of the invention, 28 where ~ is an alkenyl group having one or more 29 double bonds can be made, for example, by having the requisite number of double bonds incorporated into 31 the reagent of Formula 3. Generally speaking, such 32 compounds where A is an unsaturated carbon chain can 33 be obtained by synthetic schemes well known to the 34 practicing organic chemist; for example by Wittig and like reactions, or by introduction of a double 36 bond by elimination of halogen from an CA 02238310 1998-0~-21 1 alpha-halo-carboxylic acid, e~ter or like 2 carboxaldehyde. Compounds of the invention where 3 the A group has a triple (acetylenic) bond can be 4 made by using the corresponding aryl or heteroaryl aldehyde intermediate. Such intermediate can be 6 obtained by reactions well known in the art, for 7 example, by reaction of a corresponding methyl 8 ketone with strong ba~e, such as lithium diisopropyl 9 amide.
o The acids and salts derived from compounds of Formula 1 are readily obtainable from the 12 corresponding esters. Basic saponification with an 13 alkali metal base will provide the acid. For 14 example, an ester of Formula 1 may be di~;solved in a polar solvent such as an alkanol, preferably under 16 an inert atmosphere at room temperature, with about 17 a three molar exces~ of base, f~or F-x~mple, potassium 18 or lithium hydroxide. The solution is stirred Eor 19 an extended period of time, between 15 and 20 hours, cooled, acidified and the hydrolysate recovered by 21 conventional means.
~ The amide (in Formul~ 1 B is CONRgRlo) may be 23 formed by any appropriate amidation means known in 24 the art from the corresponding esters or carboxylic 2~s acids. One way to prepare such compounds is to 26 convert an acid to an acid chloride and then treat 27 that compound with ammonium hydroxide or an 28 appropriate amine.
29 Alcohols are made by converting the corresponding acids to the acid chloride with 31 thionyl chloride or other m~;lnS (J. March, "Advanced ~2 Organic Chemistry" r 2nd Edition, McGraw-Hill Book 33 Company), then reducing the acid chloride with 34 sodium borohydride (March, Ibid, pg. 1124), which gives the corresponding alcohols. Alternatively, 36 esters may be reduced with lithium aluminum hydride CA 02238310 1998-0~-21 W O 97/19062 PCT~US96/18529 -1 at reduced temperatures. Alkylating these alcohols 2 with appropriate alky halides under Williamson 3 reaction conditions (~larch, Ibidr pg. 357~ gives the 4 corresponding ethers. These alcohols can be 5 converted to esters by reacting them with 6 appropriate acids in the presence of acid catalysts 7 or dicyclohexylcarbodiimide and ~ dimethylaminQpyr~d7ne.
9 A~dehydes can be prepared from the corresponding primary alcohols using mild oxidi2ing agents such as 11 pyridinium dichromate in methylene chloride (Corey, 12 E. 3., Schmidt, G., Tet. Lett., 399, 1979), or 13 dimethyl sulfoxide/oxalyl chloride in methylene 14 chloride (omura~ K., Swern, D., Tetrahedron, 1978 r 34, 1651).
16 Ketones can be prepared from an appropriate 17 aldehyde by treating the aldehyde with an alkyl 18 Grignard reagent or similar reagent followed by 1~ oxidation.
Acetals or ketals can be prepared from the 21 corresponding aldehyde or ketone by the method 22 described in March, Ibid, p 81~.
23 Compounds of FOrmU1a 1 where B is H can be 24 prepared from the corresponding halogenated aromatic 25 compounds, preferably where the halogen is I.

CA 022383l0 l998-05-2l W O 97/l9062 PCTAUS96/18529 4 ~ 3)E~yl-Ym~-2nuoo benzoate (Compound G~
s~tt N tsu Pyrtdine CH~Ch 6 Comi?OUlld C BU Compound I
P ~C02EI

~ 1) SOC17 , \~J~NJ~
tt2 8ut 1 ~1 21 E~tyl 4-~li t N~J

13 Compound C tsu Compound 3 ~ 3)E~y)4ro~o-2-~o~ r~ ~ ~ J C4 18 benzoate ~Compound G) sut tsu Pyridine CH?CI-19 su Compound 5 2t 23 sut~co2H EDC.Di~L~P But~ J~M N~Co2Et l Ethyi 4-t~mino-2-fiuoro I _ H
~ ~ oenzoate (Compound G) 26 '~ Pyrtdine CH.CI. \T~

28 Compound F tsu Compound N

~ ~ SF~ OEI. ~ N ~ 42 35 tsu Compound N tsu Compound 7 36 Reaction Scheme 5 WO 97/19062 PCT~S96/18S29 S ~"CO~H ~ SOCI ~N ~ ' 7 ~ 2) Cnmro~m~ H F
N~ 3)NaOH~tOH N~

(~nmro~ 1 10 11 rnmrolmri C F

4 ~/ 1) SOCI2 ~ ~ ~ NJ~ ~ o~2H
I ¦ 2) t'nmrmm~l I/Py H
~ 3)NaOHVEtOH ~ ~

18 C~mpo~d12 23 ~_~ ~ l) SOCI~ ~f ~NJ~ cc2H
28 l l ~)~nmroln~lK~Py l l H N02 N~ 3)NaOH~OH N~
('nm~- n~l 14 ~1 f'nmr~ nrl C

36 . Reaction Scheme 5 (continued~

CA 022383l0 l998-0~-2l 1 Reaction Scheme 5 illustrates examples for the - 2 formation of the carbamoyl (amide) compounds of the 3 present invention by reactlon of a reagent of 4 Formula 2 with a reagent of Formula 3. Thus, 5 2,6-di-tert-butylisonicotinic acid (Compound C) is 6 reacted with thionyl chloride (SOC12) to provide the 7 intermediate acid chloride, which is then reacted ~ with ethy~. ~-fJ.uoro-4-amino-benzoate (Com~ound G~ in g the presence of an acid acceptor (pyridine) to yield ~o ethyl 2-fluoro-4-[(2'6'-di-tert-butylpyrid-4~-11 yl)carbamoyl]benzoate (Compound 1). As another 12 example, 3,5-di-tert-butylbenzoic acid (available by 13 the literature procedure of Kagechika et al., J.
14 Med. Chem. 198~, 31, 2182, incorporated herein by 15 reference) is reacted with thionyl chloride, 16 followed by ethyl 2-:Eluoro-4-amino-benzoate 17 (Compound G) to yield ethyl 18 2-fluoro--4-[(3',5'--di-tert-butylphenyl)carbamoyl~ben 19 zoate ~Compound 5). As still another example, 20 0-methoxymethyl-3,5-di-tert-butylsalicylic acid 21 ( Compound F)is reacted with ethyl 22 2-fluoro-4-amino-benzoate (Compound G) in the 23 presence of 4-dimethylaminopyridine (DMAP~ catalyst 24 and ~-(3-dimethylaminopropyl)-3-ethylcarbodiimide 25 hydrochloride (EDC) to give ethyl 26 2-fluoro-4-~(2'-methoxymethyl-3',5'-di-tert-butylphe 27 nyl)carbamoyl]benzoate (Compound N). The 28 methoxymethyl protecting group is removed from 29 Compound N by treatment with borontrifluoride 30 ethereate and thiophenol to yield ethyl 31 2-fluoro-4-~(2'-hydroxy-32 3',5'-di-tert-butylphenyl)carbamoylJbenzoate 33 ( Compound 7).
~ In yet another example shown in Reaction Scheme 35 5, 2,6-di-tert-butylisonicotinic acid (Compound C) 36 iS reacted with thionyl chloride (SOCl2), the W O 97/19062 PCT~US96/18529 1 resulting intermediate acid chloride is reacted with 2 methyl 2~6-difluoro--4-amino benzoate (Compound I), 3 followed by saponification of the ester group, to 4 yield 2,6-difluoro-4-[~2',6'-di-tert-butylpyrid-5 4'yl)carbamoyl~benzoic acid (Compound 10).
6 3,5-Di-tert-butylbenzoic acid is subjected to the 7 same sequence of reactions to provide 8 2 ~ 6-dif J uoro-4~ 3 ~ r 5'-di-tert-butylphenyl~carbamoyl 9 ] benzoic acid (Compoun~a 12).
0 As yet another exaLmple, shown in Reaction Scheme 11 5, 2,6-di-tert-butylisonicotinic acid (Co~pound C) 12 iS reacted with thionyl chloride (SOCl2), followed by 1S methyl 2-nitro-4-aminobenzoate ( r~ und K) and 14 saponification of the ester function to give 15 2-nitro-4-[(2',6'-di-tert-butylpyrid-4'-yl)carbamoyl 16 ] benzoic acid (Compound 14).
17 ~peci fic 18 ~Y~mrles4-Bromomethyl-2,6-di-t-butylpyridine 1 ~ ( Con~r~und A) To a mixture of 2,6-di-t-butyl-4-methylpyridine 21 (Aldrich, 2.0 g, 9.73 mmol) in 25 ml of dry CCl4 wa~
~ ~dded benzoyl peroxide (24 mg, 0.097 mmol) and NBS
23 (1. 9 g, 10.7 mmol). The reaction mixture was 24 refluxed for 16 hours. After it cooled to room 25 temperature, the solvent was removed in vacuo and 26 the residue was purified by column chromatography 27 (silica gel, hexane) to give an oil (1.957 g) which 28 contained 8296 of the desired product and 1896 of the 29 starting material. lH NMR ~; 7.09 (s, 2H), 4.39 (s, 30 2H), 1.35 (s, 18H).
31 4-Hydroxymethyl--2,6-di--t--butylpyridine (Compound B) 32 A heterogeneous solution of 33 4-bromomethyl-2,6-di-t--butylpyridine (Compound A, 34 1. 743 g, 8296 purity) in 20 ml of 1296 NaOH in water 3~ and 10 ml of 1,4-dioxane was refluxed for 12 hours.
36 The solution spontaneously separated into two 12lyers CA 02238310 1998-0~-21 W O 97/19062 PCTAUS96/~8529 1 as it cooled to room temperature. The upper layer 2 was separated and ethyl acetate was added. This 3 organic layer was then washed with }:~rine, water and 4 dried over MgSO4. The desired product was purified 5 by column chromatography (ethyl acetate/hexane 1/9) 6 to give a white solid. lH NMR ~i 7.09 (s, 2H), 4.67 7 (d, J = 4.4 Hz, 2H), 2.3 (b, lH), 1.36 (s, 18H).
8 2,6-Di-t-butylisonicotinic acid (~ _--uIld C~
~ Jone's reagent was added dropwise to a solution of 4-hydroxymethyl-2,6-di-t-butylpyridine (Compound 11 B, 302 mg, 1.37 mmol) in 5 ml of acetone until the 12 solution changed color from light yellow to orange 13 (55 drops of Jone's reagent were consumed). After 5 14 minutes 2 ml of isopropanol were added to the 15 reaction mixture, and a green precipitate of Cr3+
t6 salt was formed. The precipitate was removed by 17 ~iltration and the solution was diluted with ethyl 18 acetate, then washed with brine, water and dried 19 over MgSO4. After filtration, the solvent was 20 removed to give the desired product as a white solid 21 (227 mg) lH NMR ~ 7.71 (s, 2H), 1.34 (s, 18H).
22 2-Bromo-4,6-di-t-butylphenol (Compound D) 23 To a solution of 2,4-di-t-butylphenol (Aldrich, 24 2.0 g, 9.7 mmol) in 2 ml of HOAc was added Br2 (~ ~
25 ml, 9.7 mmol). The reaction mixture was stirred at 26 room temperature for 12 hours. Solvent was removed 27 under reduced pressure and the residue was purified 28 by column chromatography (ethyl acetate/hexane 1/20) 29 to yield the desired product (2.54 g) as a white 30 solid. 1H NMR ~ 7.33 (d, J = 2.3 Hz, lH), 7.24 (d, J
31 = 2.3 Hz, lH3, 1.41 (s, 9H), 1.29 (s, 9H).
32 O-Methoxymethyl-2-bromo-4~6-di-t-butylphenol 33 ( Compound E) 34 To a solution of 2-bromo-4,6-di-t-butylphenol 35 (Compound D 2.54 g, 8.88 mmol) and catalytic amoun~
36 of Bu4NI in 20 ml of dry CH2C12 at 0~C was added CA 022383l0 l998-05-2l W O 97/19062 PCT~US96/18529 1 diisopropylethylamine (9.51 ml, 53 mmol), followed 2 by methoxymethyl chloride (2.02 ml, 26.6 mmol). The 3 reaction mixture was heated to 4~~C for 12 hours.
4 The reaction mixture was then washed with 10% citric acid, then NaHCO3 (sat.), brine, and dried over 6 MgSO4. After filtration and removal of the solvent 7 under reduced pressure, the residue was puri~ied by 8 co~umn chromatograp~y ~l?ure hex~ne) to yield the title compound ~2.79 g) as a colorless oil. lH NMR
0 7.40 (d, J = 2.44 Hz, lH), 7.30 (d, J = 2.4 Hz, lH), 5.22 (s, 2H), 3.70 (s, 3H), 1.43 (s, 9H), 1.29 (s, 2 9H).
3 O-Methoxymethyl-3',5'-di--t-butylsalicylic acid 4 ( ~O. ~und F) To a solution of O-methoxymethyl-2-bromo-4,6-16 di-t--butylphenol (~ und ~, 2.79 g, 8.5 mmol) in 7 30 ml of dry THF at -78~C under Ar was added 11 ml 8 of t--BuLi (1.7 M in hexane, 18.7 mmol). This 1~ mixture was stirred at -78~C for 1 hour. Then CO2 20 (g) was bubbled into the solution at --78~C for 1 21 hour. After removal o:E the CO2 stream, the reaction 22 mixture was stirred for an additional hour at -78~C.
23 Then 10% of HCl was added and the mixture was 24 allowed to warm to room temperature and extracted 25 with ethyl acetate. The organic layer was washed 26 with brine and dried over Na2SO4. After 27 concentration, the residue was purified by column 28 chromatography (ethyl acetate/hexane 1/1) to yield 29 the title compound as a white solid (492 mg). l~I NMR
30 ~; 7.75 (d, J = 2.81 Hz, lH), 7.60 (d, J = 2.8 Hz, 31 lH), 5.07 (s, 2H), 3.62 (s, 3H), 1.33 (s, 9H), 1.26 32 (S, 9H). EthYl 4--Amino-2-fluorobenzoate (Compound G) 33 To a mixture of 2--fluoro--4--nitrotoluene(1.0 g, 34 6.4 mmol, Aldrich) and Na2Cr2O~ (2.74 g, 8.4 mmol) in 35 13.7 ml of HOAc was added slowly 6.83 ml of H2SO4.
36 This mixture was slowlyr heated to 90 ~C for 1 hour to CA 022383l0 l998-0~-2l 1 give a greenish heterogeneous solution. The mixture 2 was cooled to room temperature and diluted with 3 ethyl acetate. The pH of the sOlution was adjusted 4 to 4 with aqueous NaOH. The mixture was extracted with more ethyl acetate. The combined organic 6 layers were washed with NaHCO3(sat.), then brine and 7 dried over Na2SO4. After filtration, the solution 8 was Goncentrated to dryness which then was dissolved g in 6 ml of SOCl2, and heated at 80 ~C for 1 hour.
The excess of SOCl2 was removed under reduced pressure and the residue was dissolved in 5 ml of 2 CH2C12, 2 ml of EtOH and 2 ml of pyridine. The 3 mixture was stirred at room temperature for 2 hours 4 and concentrated to dryness. Ethyl 2--fluoro-4-nitrobenzoate was obtained as a white 6 solid after column chromatography of the residue 7 with ethyl acetate/hexane (1/9). This solid was 8 then dissolved in 10 ml of ethyl acetate, and Pd/C
19 ( 50 mg) was added. Hydrogenation converted ethyl 2-fluoro-4-nitrobenzoate into the title compound.
21 lH NMR ~; 7.77 (t, J -- 8.4 E~z, lH), 6.41 (dd, Jl =
22 8.~, J2 = 2.2 Hz, lH), 6.33 (dd, J1 = 13.0, J2 = 2.2 23 Hz, lH), 4.33 (q, J = 7.1 Hz, 2H), 4.3 (b, 2H~, 1.37 24 (t, J = 7.1 Hz, 3H).
~ethyl 4--Amino-2,6-difluorobenzoate (Compound I~
26 A solution of trifluorobenzoic acid (150 mg, 27 O. 85 mmol, Aldrich) in 0.5 ml of SOC12 was heated 28 under reflux for 2 hours. The reaction mixture was 29 cooled to room temperature, and excess of SOCl2 was removed under reduced pressure. The residue was 31 dissolved in 1 ml of pyridine and 0.2 ml of 32 methanol. After stirring at room temperature for 30 33 min, solvent was removed and the residue was 34 purified by column chromatography (ethyl acetate/hexane 1/10) to give methyl 36 trifluorobenzoate as a colorless oil. This oil was CA 022383l0 l998-05-2l 1 then dissolved in 1 ml o~ CH3CN, then a solution o~
2 NaN3 (100 mg, 1.54 mmo.L) in 0.5 ml o~ water was 3 added. The reaction m.ixture was refluxed for two 4 days. Salt was removed ~y filtration and the 6 remaining solution was concentrated to an oil. This 6 oil was then dissolved in 1 ml o:~ methanol, followed 7 by a catalytic amount of Pd/C (10%, w/w). The 8 reacti.on mixtu.re was hydxogenated for 12 hours~
g Catalyst was removed and the solution was concentrated to an oil. After column chromatography 11 (ethyl acetate/hexane 1/3), the title compound was 12 obtained as colorless crystals.
13 lH NMR ~ 6.17 (d, J = 10.44 Hz, 2H), 4.2 (b, 2H), ~4 3.87 (s, 3H).
Methyl 2-Nitro-4-aminobenzoate (Co~npound K) 16 2--Nitro--4--aminobenzoicacid (261 mg, 1.43 mmol) 17 was di solved in 1 ml of SOC12. The solution was 8 refluxed for 1 hour. ]3xcess SOCl2 was removed under 19 reduced pressure and 5 ml of C~2Cl2, 1 ml of MeOH and TEA (O.24 ml, 1.7 mmol~ were added to the residue.
21 The reaction mixture w2ls stirred at room temperature for 2 hours. Excess MeOH and TEA were removed and 23 the residue was puri~ied by column chromatography 24 with ethyl acetate/hexane (1/3) to yield the title 25 compound as a yellow solid (316 mg). lH NMR ~; 7.69 26 ~d, J = 8.5 I~z, lH), 6.85 (d, J = 2.2 Hz, lH), 6.67 27 (dd, J = 8.3; 2.1 Hz, lH~, 4.31 (b, 2H~, 3.94 (s, 28 3H).
29 Ethyl 2-f luoro-4- r ( 2' 6~-di-t-butylpyrid-4~-30 yl~ carbamoyllbenzoate (Compound 1) 31 A solution of 2,6-,di--t--butylisonicotinic acid 32 ( Compound C, 47.3 mg, 0.20 mmol) in 2 ml of SOC12 was 33 heated under reflux for 2 hours. Excess SOCl2 was 34 removed in vacuo and th.e residue was dissolved in 2 35 ml of dry CH2Cl2, and et:hyl 2-~luoro--4--amino~enzoate 36 ( Compound G, 40.2 mg, 0.22 mmol) and pyridine 1 (0.0835 ml, 0.69 mmol) were added. The reaction 2 mixture was stirred at room temperatu~e for 12 3 hours. Solvent was removed and the residue was 4 purified by column chromzltography (ethyl acetate/hexane 1/9) to yield the title compound 6 (71.2 mg) as white crystals. lH NMR ~i 8.56 (b, lH), 7 7.91 (t, J = 8.36 Hz, lH), 7.53 (ddr J = 12.82, 2.0 8 Hz, lH), 7.39 (dd, J = 8.7, 2.0 Hz, lH), 4.33 (q, J
9 = 7.1 Hz, 2H), 1.37 (t, J = 7.1 Hz, 3H), 1.35 (s, o 18H).
Ethyl 4- r ( 2~,6~-di-t-but~lpyrid-4~-yl)carbamoyll-12 benzoate (Compound 3) 13 Using the same procedure as for the synthesis of 14 ethyl 2-fluoro--4-~(2'6'--di--t-butylpyrid--4'--yl)carbamoyl]benzoate (~Y- p~und l) but using 16 2,6-di--t-butylisonicotinic acid (C:ompound C, 101 mg, 17 O. 43 mmol) and ethyl 4-aminobenzoate (78 mg, 0.47 18 mmol), the title compound was obtained as a white 19 solid (135 mg). lH NMR ~ 8.43 (b, lH),, 8.02 (d, J =
8.7 Hz, 2H), 7.75 (d, J c 8.7 Hz, 2H), 7.48 (s, 2H), 21 4.33 (q, J = 7.1 Hz, 2H), 1.38 (t, J = 7.1 Hz, 3H), 22 1.35 (s, 18H).
23 Ethyl 2-Fluoro-4- r ( 3~,5'-di-t-butylphen-24 yl) carbamoyl~benzoate (Compound 53 Using the same procedure as for the synthesis of 26 ethyl 2-fluoro-4-[(2'6'-di-t-butylpyrid-4'-27 yl)carbamoyl~benzoate (Compound 1) but using 28 3,5-di--t-butylbenzoic acid (60 mg, 0.26 mmol, 2û available by literature procedure, see Kagechika et al. J. Med Chem. 1988 31, 2182 - 2192) and ethyl 31 2-fluoro-4-aminobenzoate (Compound G, 51.5 mg, 0.28 32 mmol), the title compound was obtained as a white 33 solid (66 mg). lH NMR ~; 8.21 (b, lH), 7.93 (t, J =
34 8.3 Hz, lH)~ 7.79 (dd, J = 12.8, 2.0 Hz, lH), 7.67 (d, J = 1.8 Hz, 2H), 7.65 ~t, J = 1.7 Hz, lH), 7.35 36 (dd, J = 8.7, 2.1 Hz, lH), 4.36 (~, J = 7.2 Hz, 2H), 1 1.39 (t, J = 7.2 Hz, 3H), 1.36 (s, 18H).
2 Ethyl 2-Fluoro-4-l(2'-methoxymethyl-3',5'--di-t-3 butylphenyl)carbamoyllbenzoate (Compound N~
4 To a mixture of 0-methoxymethyl-3',5'-di-t-butylsalicylic acid 6 (Compound F, 150 mg, 0.51 mmol), 7 4-dimethylaminopyridine (142 mg, O.61 mmol) and 8 ethyl 2-fluoro-4-aminobenzoate (Compound G, 102 mgr ~ 0.56 mmol) in 5 ml of ,dry CH2C12 was added 1-(3-di-methylaminopropyl)--3--ethylcarbodiimidehydrochloride 11 (117 mg, 0.61 mmol). The reaction mixture was 12 stirred at room temperature for 12 hours. Solvent 13 was evaporated in vacuo and the residue was 14 dissolved in ethyl ace-tate, then washed with brine, water and dried over MgSO4. After filtration, 16 solvent was removed and the residue was purified by 17 column chromatography ~ethyl acetate/hexane 1/3) to 18 give the title compound (58 mg). 1H NMR ~ 8.97 (b, 19 lH~, 7.94 (t, J = 8.37 Hz, lH), 7.78 (d, J = 2.7 Hz, lH), 7.61 (d, J = 13.0 Hz, lH), 7.~i6 (d, J = 2.6 Hz, 21 lH), 7.35 (d, J = 8.7 ~z, lH), 5.00 (s, 2H), 3.53 ~ (s, 3H), 4.38 (q, J = 7.1 Hz, 2H), 1.47 (s, 9H), 23 1.39 (t, J = 7.2 Hz, 3~I), 1.33 (s, 9H).
24 Ethyl 2-Fluoro-4-r(2'-hydroxy-3',5'-di-t-butylphenYl)carbamoYl~benzoate (f'~mr~und 7) 26 To a solution of ethyl 27 2-fluoro-4-~(2~-methoxymethyl-3',5'-di-t-butylphenyl 28 )carbamoyl]benzoate (Cc~,.. nd N, 34 mg, 0.07 mmol) 29 in 1 ml of THF were added 10 drops of HOAc. The reaction mixture was heated to reflux for 12 hours.
31 Solvent was removed and ethyl acetate was added.
32 The solution was washed with NaCHO3 (sat.), brine, 33 water and dried over MgSO4. Solvent was removed in 34 vacuo to give an oil. The oil was allowed to be exposed to the atmosphere for 12 hours during which 36 time crystals ~ormed. The crystals were collected CA 022383l0 l998-0~-2l W O 97/19062 PCTAUS96/18~29 .

1 and washed several times with hexane to afford the 2 title compound as a white solid (13.5 mg). lE~ NMR
3 10.73 (s, lH), 7.98 (d, J = 2.56 Hz, lH), 7.88 (b, 4 lH), 7.75 (t, ~ = 8.26 Hz, lH), 7.60 (d, J = 2.44 6 Hz, lH), 7.32 (dd, J = 12.3, 2.0 ~z, lH), 7.02 (dd, 6 J = 8.6, 2.0 Hz, lH), 4.35 (q, J = 7.2 Hz, 2H), 1.39 7 (S, 9H), 1.37 (t, J = 7.2 Hz, 3H), 1.5 (s, 9H).
8 2,6--Difluoro--4--r ( 2',6'-di--t-butYlpyrid--4~yl)carbamoy llbenzoic Acid (Compound 10) o To 2,6-di-_-butylisonicotinic acid (Compound C, 11 20 mg, 0.085 mmol) was added 1 ml of SOCl2. The 12 mixture was heated under reflux for 2 hours. After 13 cooling to room temperature, excess SOC12 was removed 14 and the residue was dissolved in 2 ml of CH2Cl2. To 15 this solution was added methyl 16 2,6-difluoro-4-aminobenzoate (Compound I, 16 mg, 17 0.085 mmol) and triethylamine (0.015 ml, 0.1 mmol).
18 The reaction mixture was kept at room temperature 19 for 2 hours and then concentrated to dryness. The 20 residue was purified by column chromatography with 21 ethyl acetate/hexane (1/10) to yield the methyl 22 ester of the title compound. This was saponified 23 . according to the general procedure (see below) to 24 give the title compound as a colorless solid. lH N~R
25 ~ 7.44 (s, 2H), 7.40 (d, J = 11.8 Hz, 2H) 1.37 (s, 26 18H).
27 2,6-Difluoro-4- r t3',5'-di-t-butylphenyl)carbamoyllbe 28 nzoic Acid (Compound 12) 29 Using the same procedure as ~or the preparation 30 of 2~6-difluoro-4-[(2~6~-di-t-butylpyrid-31 4~yl)carbamoyl]benzoic acid (Compound 10) but using 32 3,5-di-t-butylbenzoic acid (37 mg, 0.16 mmol) and 33 methyl 2,6-difluoro-4-aminobenzoate (Compound I, 29 34 mg, 0.16 mmol), the title compound was obtained as 35 colorless crystals. lH NMR ~i 7.92 (b, lH) 7.60 (m, 36 3H), 7.42 (d, J = 10.0 Hz, 2H), 1.38 (s, 18H).

W O 97/19062 PCT~US96/18529 1 2-Nitro-4- r ( 2r,6'-di-t-butylpYrid-4'-yl)carbamoyl~be 2 nzoic Acid (Compound 14) 3 Using the same procedure as for the preparation 4 of~ 2,6-dii~luoro-4--[(2',6'--di-t--butylpyrid-4'yl)carbamoyl]benzoic acid (l~o~~round 10) but using 6 2,6-di-t-butylisonicotinic acid (40 mg, 0.17 mmol) 7 and methyl 2-nitro-4-aminobenzoate (Compound K, 33 8 mg~ Q.17 mmol~, the titl~ compound wa~ obtained as a 9 light yellow oil. 1H NMR ~ (acetone-d6) 10.25 (b, 10 lH), 8.32 (s, lH), 7.97 (d, J = 8.1 Hz, lH), 7.93 11 (b, lH), 7.70 (s, 2H), 1-36 (s~ 18H)-12 General procedure for the syntheses o~ benzoic acid 13 derivatives by hydrolyzing the corresponding methyl 14 or ethyl esters To a solution o~ ~ster (3.0 mmol) in 20 ml o~
6 EtOH was added 5 ml of 1 N NaOH in water. The 7 reaction mixture was stirred at room temperature ~or 8 overnight and neutralized with 1096 HCl to PH=5. The 19 alcohol was removed by evaporation and the aqueous 20 layer was extracted with ethyl acetate (3xlOml).
21 The ethyl acetate layer was further washed with 22 NaHCO3 (sat.), brine and dried over MgSO4. After 23 concentration, the desired carboxylic acid was 24 obtained which could be recrystallized in ethyl 25 acetate or acetonitrile.
26 2-Fluoro--4--r (2',6'-di--t-butylpyrid--4'-yl)carbamoyllb 27 enzoic Acid (Cont~ound Z) 28 lH NMR ~ (CD30D) 7.g2 (t, J = 8.36 Hz, lH), 7.82 29 (dd, J = 12.82, 2.0 Hz, lH), 7.63 (s, 2H), 7.55 (dd, 30 J = 8.7, 2.1 Hz, lH~, 1.39 (s, 18H~.
31 4- r ( 2~,6~-Di-t-butylpyrid-4~-yl)carbamoyllbenzoic 32 acid (Compound 4) 33 lH NMR ~i (CD30D) 8.02 (d, J = 8.85 Hz, 2H), 7.85 34 (d, J = 8.85 Hz, 2H), 7.63 (s, 2H), 1.40 (s, 18H).
35 2-Fluoro-4- r ( 3~,5~-di-t-butyl)phenylcarbamoyllbenzoi 36 C acid (Compound 6) W O 97/19062 PCT~US96/18S29 1 lH NMR ~ (CD30D~ 7.92 (t, J = 8.3 Hz, lH), 7.80 2 (dd, J = 12.8, 2.0 Hz, lH), 7.79 (d, J = 1.8 Hz, 3 2H), 7.69 (t, J = 1.7 Hz, lH), 7.57 ~dd, J = 8.7, 4 2.1 Hz, lH), 1.37 (s, 18H).
2-Fluoro-4- r ( 2~-hydroxy-3~,5~-di-t-butyl)phenylcarba 6 moyl~benzoic acid (Compound 8) 7 lH NMR ~; (acetone-d6) 12.3 (b, lH), 10.07 (b, 8 lH~, 7 98 ~t, J = 8.48 Hz~ lH)~ 7.80 (m, 2H~, 7.58 g (d, J = 2.3 Hz, lH), 7.56 (dd, J = 8.8, 2.0 Hz, lH), o 1.44 (s, 9H), 1.31 (s, 9H).

Claims

WHAT IS CLAIMED IS:
1. A compound of the formula wherein X is CH or N;
R1 is independently H or alkyl of 1 to 6 carbons;
m is an integer having the value of 0 - 5;
p is an integer having the value of 0 - 2;
r is an integer having the value 0 - 2;
L is -(C=Z)-NH- or -NH-(C=Z)- where Z is O or S;
Y is a phenyl or naphthyl group, or heteroaryl selected from a group consisting of pyridyl, thienyl, furyl, pyridazinyl, pyrimidinyl, pyrazinyl, thiazolyl, oxazolyl,imidazolyl and pyrrazolyl, said phenyl, naphthyl and heteroaryl group being optionally substituted with one or two R1 groups;
W is a substituent selected from the group consisting of F, Br, Cl, I, C1-6alkyl, fluoro subtituted C1-6 alkyl, NO2, N3, OH, OCH2OCH3, OC1-10alkyl, retrazol, CN, SO2C1-6-alkyl, SO2C1-6alkyl, SO2C1-6-fluoro substituted alkyl, SO-C1-6 alkyl, CO-C1-6alkyl, COOR6, phenyl, phenyl CLAIM

itself substituted with a W group other than with phenyl or substituted phenyl with the provisos that when r is 0, or when Y is phenyl and (W)r represent OH then p is not 0 and at least one W in the ring having the X group is not selected from the group consisting of C1-6alkyl, OH and OC1-10alkyl;
A is (CH2)q where q is 0-5, lower branched chain alkyl having 3-6 carbons, cycloalkyl having 3-6 carbons, alkenyl having 2-6 carbons and 1 or 2 double bonds, alkynyl having 2-6 carbons and 1 or 2 triple bonds, and B is COOH or a pharmaceutically acceptable salt thereof, COOR8, CONR9R10, -CH2OH, CH2OR11, CH2OCOR11, CHO, CH(OR12)2, CHOR13O, -COR7, CR7(OR12)2, CR7OR13O, where R7 is an alkyl, cycloalkyl or alkenyl group containing 1 to 5 carbons, R8 is an alkyl group of 1 to 10 carbons or trimethylsilylalkyl where the alkyl group has 1 to 10 carbons, or a cycloalkyl group of 5 to 10 carbons, or R8, is phenyl or lower alkylphenyl, R9 and R10 independently are hydrogen, an alkyl group of 1 to 10 carbons, or a cycloalkyl group of 5-10 carbons, or phenyl or lower alkylphenyl, R11 is lower alkyl, phenyl or lower alkylphenyl, R12 is lower alkyl, and R13 is divalent alkyl radical of 2-5 carbons.
2. A compound in accordance with Claim 1 wherein L is -(C=Z)-NH-.
3. A compound in accordance with Claim 1 wherein L is -HH-(C=Z)-.
4. A compound in accordance with Claim 1 wherein X is CLAIM

CH.
5. A compound in accordance with Claim 1 wherein X is N.
6. A compound in accordance with Claim 1 wherein Y is selected from the group consisting of phenyl, pyridyl, 2-thiazolyl, thienyl and furyl. 7. A compound in accordance with Claim 6 wherein Y is phenyl.
8. A compound in accordance with Claim 1 wherein W is branch-chained alkyl,F, NO2, Br, I, CF3, N3, or OH.
9. A compound in accordance with Claim 8 wherein (W)p represents two branch-chained alkyl groups.
10. A compound in accordance with Claim 8 wherein (W)r represents one or two fluoro or one or two NO2 groups.
11. A compound of the formula wherein X is CH or N;
R1 is independently H or alkyl of 1 to 6 carbons;
m is an integer having the value of 0 - 5;

CLAIM

p is an integer having the value of 0 - 2;
r is an integer having the value 0 - 2;
Z is O or S;
W is a substituent selected from the group consisting of F, Br, Cl, I, C1-6alkyl, fluro substituted C1-6alkyl, NO2, N5, OH, OCH2OCH3, OC1-6alkyl, tetrazol, CN, SO2C1-6-alkyl, SO2C1-6-alkyl, SO2C1-6-fluoro substituted alkyl, SO-C1-6 alkyl, CO-C1-6alkyl, COOR8, phenyl, phenyl itself substituted with a W group other than with phenyl or substituted phenyl with the provisos that when r is 0, or when Y is phenyl and (W)r represent OH then p in not 0 and at least one W in the ring having the X group is not selected from the group consisting of C1-6alkyl, OH and OC1-10alkyl;
A is (CH2)q where q is 0-5, lower branched chain alkyl having 3-6 carbons, cycloalkyl having 3-6 carbons, alkenyl having 2-6 carbons and 1 or 2 double bonds, alkynyl having 2-6 carbons and 1 or 2 triple bonds, and B is COOH or a pharmaceutically acceptable salt thereof, COOR~, CONR9R10, -CH2OH, CH2OR11, CH2OCOR11, CHO, CH(OR12)2, CHOR13O, -COR7, CR7(OR12)2, CR7OR13O, where R7 is an alkyl, cycloalkyl or alkenyl group containing 1 to 5 carbons, R~ is an alkyl group of 1 to 10 carbons or trimethylsilylalkyl where the alkyl group has 1 to 10 carbons, or a cycloalkyl group of 5 to 10 carbons, or R8 is phenyl or lower alkylphenyl, R9 and R10 independently are hydrogen, an alkyl group of 1 to 10 carbons, or a cycloalkyl group of 5-10 carbons, or phenyl or lower alkylphenyl, R11 is lower alkyl, phenyl or lower alkylphenyl, R12 is lower alkyl, and R13 is divalent alkyl radical of 2-5 carbons.
12. A compound in accordance with Claim 11 wherein X is CH.
13. A compound in accordance with Claim 12 wherein Z is O.
14. A compound in accordance with Claim 13 wherein A is (CH2)q, and B is COOH or a pharmaceutically acceptable salt thereof, COOR8 OR CONR9R10.
15. A compound in accordance with Claim 14 wherein (W)p represents two branch-chained alkyl groups and (W)~ represents one or two fluoro or one or two NO2 groups.
16. A compound in accordance with Claim 15 wherein the phenyl ring is 1,4-substituted by the (C=Z)NH- and -A-B- groups.
17. A compound of the formula wherein X is CH or N;

CLAIM

W1 is H or OH;
W2 is H, F or NO2;
W3 is H, F or NO2;
R~ is H, CH3 or C2H5, with the proviso that when X is CH then W1, W2 and W3 all are not H and at least one of W1 and W2 is selected from the groups consisting of F and NO2.
18. A compound in accordance with Claim 17 wherein X is N.
19. A compound in accordancc with Claim 18 wherein W2 is F and W3 is H.
20. A compound in accordance with Claim 19 which is:
ethyl 2-fluoro-4-[(2'6'-di-t-butylpyrid-4'-yl)carbamoyl]benzoate, or 2-fluoro-4-[(2'6'-di-t-butylpyrid-4'-yl)carbamoy]bezoic acid.
21. A compound in accordance with Claim 18 wherein W2 is F and W3 is F.
22. A compound in accordance with Claim 21 which is:
methyl 2,6-difluoro-4-[(2',6'-di-t-bytylpyrid-4'yl)-carbamoyl]benzoate, or 2,6-difluoro-4-[(2',6'-di-t-bytylpyrid-4'yl)carbamoyl]benzoic acid.
23. A compound in accordance with Claim 18 wherein W3 is NO2.
24. A compound in accordance with Claim 23 which is:
methyl 2-nitro-4-[(2',6'-di-t-bytylpyrid-4'-yl)carbamoyl]benzoate, or 2-nitro-4-[(2',6'-di-t-bytylpyrid-4'-yl)carbamoyl]benzoic acid.

Ch~IM ~PLA(~;EI'IT SHE~T
~3 ~ i. A co~pound m accordance u~ilh C~aim 1~ whe~ein Wl, W.
~nd W3 alC ~ hydrogen.
. A compound in accordanc_ with Claim ~ which is;
et~1 4-~2',6'-di-~ Lyl~ylid-4'- yl)carbamoyl]~en2~0ate, o~
(2',6'-di-t-buty~lpyrId-4'- yl~cal~amoy~b-nz3ic acid.
~7. A compound i~l accoldance ~ith Cla~m 17 ~herein X is i~,. A compound iI~ ac~o,~dan,~e with Ciaim ~7 whel~in W~ is and W, ~s H.
~ g. ~ co~l?c,und in accoTdance with Claim 28 ~hich is:
ethyl 2-flu~Ic,-4-[(~ '-di-t-~utylpherl-yl)ca~bamo~ en~:~ate, c ~ -fluo~o4-[(3',~ t=~ut~l)pheIIy~-car'~amoyl~benzoic a,~id.
30. A compourld in acco~dance with Claim 27 whe~ein W~ is F and W3 iS F
31. ~ compourld in acco~da~cc with Claim 30 which is methyl ~6-difluoro-~(3',~'-di-t-b~tyl-phenyl)ca~Pn~,y~ 70::~t~
2,6-difluoro4-[(3',~'-di-~-bu~yI-phc~yl)car~amoyI]benz~ic acid.
32. A compound in accoldance with Claim ~7 wherem Wl ~s 0~
33. ~ compound in accoIdance with C~laim 37 which i~:
ethyl 2-fluolo-4-[(~'-hyd~oxy-~',5'-di-~-bu~l-phenyl~carbamoy1Jl:~en~oa~c7 or ~-lfuolo~-[(~'-hyd~ox~ '-di-~-butyl)phcIIyl~arbamoyl~benzoic acId.