WO2000062808A2 - Cobalamin conjugates useful as antitumor agents - Google Patents
Cobalamin conjugates useful as antitumor agents Download PDFInfo
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- WO2000062808A2 WO2000062808A2 PCT/US2000/010100 US0010100W WO0062808A2 WO 2000062808 A2 WO2000062808 A2 WO 2000062808A2 US 0010100 W US0010100 W US 0010100W WO 0062808 A2 WO0062808 A2 WO 0062808A2
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F15/00—Compounds containing elements of Groups 8, 9, 10 or 18 of the Periodic System
- C07F15/06—Cobalt compounds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/009—Neutron capture therapy, e.g. using uranium or non-boron material
- A61K41/0095—Boron neutron capture therapy, i.e. BNCT, e.g. using boronated porphyrins
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
- A61K47/55—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds
- A61K47/551—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds one of the codrug's components being a vitamin, e.g. niacinamide, vitamin B3, cobalamin, vitamin B12, folate, vitamin A or retinoic acid
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Definitions
- Boron neutron capture therapy is based on the nuclear reaction that occurs when a stable isotope, 10 B, is irradiated with low energy (0.025 eN) or thermal neutrons to yield helium nuclei ( ⁇ -particles) and 7 Li nuclei.
- BNCT neoplasmic plasminogen activator
- 10 B is the most attractive for the following reasons: (a) it is nonradioactive and readily available, comprising approximately 20% of naturally occurring boron: (b) the particles emitted by the capture reaction [ 10 B(n, ⁇ ) 7 Li] are largely high LET: (c) their path lengths are approximately 1 cell diameter (10-14 ⁇ m), theoretically limiting the radiation effect to those tumor cells that have taken up a sufficient amount of 10 B and simultaneously sparing normal cells and (d) the extensive chemistry of boron is such that it can be incorporated into a multitude of different chemical structures.
- Li and -particles are the primary fission product of the neutron capture reaction with 10 B.
- ⁇ -Particles are relatively slow and give rise to closely spaced ionizing events that consist of tracks of sharply defined columns. They have a path length of approximately 10 ⁇ m, are high LET, and destroy a wide variety of biologically active molecules including DNA, RNA, and proteins. For these reasons there is little, if any, cellular repair from ⁇ -particle-induced radiation injury.
- boron compounds to be used for BNCT should have a high specificity for malignant cells with concomitantly low concentrations in adjacent normal tissues and blood. Since it is desirable to confine the radiation solely to these cells, an intracellular and optimally intranuclear localization of boron would be preferred.
- p- Boronophenylalanine is another compound that is being studied as a potential capture agent for the treatment of melanoma.
- the rationale for its use is the avidity of melanomas for aromatic amino acids and their subsequent incorporation into melanin (Ichihashi, M. et al., J. Invest. Dermatol..
- Tumor localization has been demonstrated following i.v. administration by means of whole body autoradiography (Coderre, J.A. et al., Cancer Res., 48, 6313-6316 (1988)) and in several patients with cutaneous melanoma following perilesional injection (Mishima, Y. et al., Sthralenther. Onkol., 165. 251-254 (1989)).
- porphyrins Two other classes of compounds with a propensity for localizing in malignant tumors are the porphyrins and the related phthalocyanines.
- the biochemical basis by which these compounds achieve elevated concentration in malignant tumors is unknown, but this observation has served as the rationale for the use of hematoporphyrin derivative in the photodyamic therapy of cancer (Dougherty, TJ. et al, Porphyrin Photosensitization, 3-13, New York: Plenum Publishing Corp. (1981)).
- the high concentration of these compounds in tumors and their intracellular localization and persistence have stimulated several groups of investigators to synthesize boronated porphyrins (Kahl, S. B.
- boron-containing purines and pyrimidines and their nucleosides are boron-containing purines and pyrimidines and their nucleosides.
- the rationale for their development is that such compounds may be selectively incorporated into rapidly proliferating tumor cells and trapped within the cell following their conversion to the corresponding nucleotide.
- these bases and their nucleosides may function as analogues of naturally occurring precursors of nucleic acids and become incorporated into nuclear DNA.
- Cytoplasmic or preferably a nuclear localization of all of these boron compounds would be advantageous since the heavy particles resulting from the capture reaction would deliver a greater proportion of their energy to intranuclear targets, thereby permitting lower boron concentrations than would have been required if the compounds were located extracellularly (Gabel, D. et al, Radiat. Res., Ill, 14-25 (1987); Fairchild, R.G. et al., Tnt. J. Rari t. Oncol. Biol. Phys.. 11, 831-840 (1985)). Schinazi and Prusoff (Schinazi, R.F.
- Figure 1 The structure of these various forms is shown in Figure 1, wherein X is CN, OH, CH 3 or adenosyl, respectively.
- cobalamin will be used to refer to all of the molecule except the X group.
- the fundamental ring system without cobalt (Co) or side chains is called corrin and the octadehydrocorrin is called corrole.
- Figure 1 is adapted from The Merck Index, Merck & Co. (11th ed. 1989), wherein X is above the plane defined by the corrin ring and nucleotide is below the plane of the ring.
- the corrin ring has attached six amidoalkyl (H 2 NC(O)Alk) substituents, at the 2, 3, 7, 8, 13, and 18 positions, which can be designated a-e and g, respectively.
- Methylcobalamin serves as the cytoplasmic coenzyme for 5 N- methyltetrahydrofolate:homocysteine methyl transferase (methionine synthetase, EC 2.1.1.13), which catalyzes the formation of methionine from homocysteine.
- Adenosylcobalamin is the mitochondrial coenzyme for methylmalonyl CoA mutase (EC5.4.99.2) which interconverts methylmalonyl CoA and succinyl CoA.
- vitamin B 12 all forms of vitamin B 12 (adenosyl-, cyano-, hydroxo-, or methylcobalamin) must be bound by the transport proteins, Intrinsic Factor and Transcobalamin II to be biologically active. Specifically, gastrointestinal absorption of vitamin B 12 relies upon the intrinsic factor-vitamin B ]2 complex being bound by the intrinsic factor receptors in the terminal ileum. Likewise, intravascular transport and subsequent cellular uptake of vitamin B 12 throughout the body is dependent upon transcobalamin II and the cell membrane transcobalamin II receptors, respectively. After the transcobalamin Il-vitamin B 12 complex has been internalized, the transport protein undergoes lysozymal degradation, which releases vitamin B 12 into the cytoplasm.
- vitamin B 12 can then be interconverted into adenosyl-, hydroxo-, or methylcobalamin depending upon cellular demand. See, for example, A.E. Finkler et al., Arch. Biochem. Biophys.. 12Q, 79 (1967); C. Hall et al, J. Cell Physiol.. 132, 187 (1987); M.E. Rappazzo et al., J. Clin. Invest, 51, 1915 (1972) and R. Soda et al, Blood, £5, 795 (1985).
- methylcobalamin is directly involved with methionine synthesis and indirectly involved in the synthesis of thymidylate and DNA, it is not surprising that methylcobalamin as well as Cobalt-57-cyanocobalamin have also been shown to have increased uptake in rapidly dividing tissue (for example, see, B.A. Cooper et al., Nature, 121, 393 (1961); H. Flodh, A ta Radiol. Snpp1. : 2S4, 55 (1968); L. Bloomquist et al., Experientia, 25, 294 (1969)).
- Vitamin B 12 has several characteristics which potentially make it an attractive in vivo tumor therapeutic agent. Vitamin B 12 is water soluble, has no known toxicity, and in excess is excreted by glomerular filtration. In addition, the uptake of vitamin B 12 could potentially be manipulated by the administration of nitrous oxide and other pharmacological agents (D. Swanson et al., Pharmaceuticals in Medical Imaging, MacMillan Pub. Co., NY (1990) at pages 621-628). A process for preparing 125 I-vitamin B 12 derivatives is described in
- the invention provides a compound of the invention which is a residue of a compound of formula I ( Figure 1) 1 linked to a residue of a molecule comprising Boron-10 (i.e., B-10), wherein X is CN, OH, CH 3 , adenosyl, or a molecule comprising B-10; or a pharmaceutically acceptable salt thereof.
- Figure 1 a residue of a compound of formula I ( Figure 1) 1 linked to a residue of a molecule comprising Boron-10 (i.e., B-10), wherein X is CN, OH, CH 3 , adenosyl, or a molecule comprising B-10; or a pharmaceutically acceptable salt thereof.
- each R is independently H or (C r C 6 )alkyl; or a pharmaceutically acceptable salt thereof.
- the invention also provides a compound wherein a residue of a compound of formula I ( Figure 1) 1 linked to a detectable radionuclide and also linked to a residue of a molecule comprising Boron-10 (i.e., B-10), wherein X is CN, OH, CH 3 , adenosyl, or a molecule comprising B-10; or a pharmaceutically acceptable salt thereof.
- the invention also provides a compound wherein a residue of a compound of formula I ( Figure 1) is linked to a detectable radoinuclide; and is also linked to a group comprising Gd-157; wherein X is CN, OH, CH 3 , adenosyl, a molecule comprising B-10, or a group comprising Gd-157; or a pharmaceutically acceptable salt thereof.
- the invention also provides a pharmaceutical composition
- a pharmaceutical composition comprising a compound of the present invention and a pharmaceutically acceptable carrier.
- the invention also provides a method of treating a tumor in a mammal in need of such treatment comprising administering to the mammal an effective amount of a compound of the present invention; and administering neutron capture therapy.
- the invention also provides a method for imaging a tumor in a mammal in need of such imaging comprising administering to the mammal a detectable amount of a compound of the present invention in combination with a pharmaceutically acceptable vehicle effective to image the tumor; and detecting the presence of the compound.
- the invention also provides a compound of the present invention for use in medical therapy or diagnosis.
- the invention also provides the use of a compound of the present invention for the manufacture of a medicament for imaging a tumor in a mammal (e.g., a human).
- the invention also provides the use of a compound of the present invention for the manufacture of a medicament for treating a tumor in a mammal (e.g., a human).
- a mammal e.g., a human
- the invention also provides intermediates disclosed herein that are useful in the preparation of the compounds of the present invention as well as synthetic methods useful for preparing the compounds of the invention.
- Figure 1 depicts the structure of cobalamin wherein X is CN
- Figure 2 illustrates the synthesis of a cyanocobalamin-nidQ- carborane conjugate.
- FIG. 3 illustrates the synthesis of representative compounds of the present invention.
- cobalamin analogs are useful, in combination with neutron capture therapy, to treat tumors.
- cobalamin analogs are useful as neutron capture agents and as tumor imaging agents.
- halo is fluoro, chloro, bromo, or iodo.
- Alkyl, alkoxy, alkenyl, alkynyl, etc. denote both straight and branched groups; but reference to an individual radical such as "propyl” embraces only the straight chain radical, a branched chain isomer such as "isopropyl” being specifically referred to.
- Aryl denotes a phenyl radical or an ortho-fused bicyclic carbocyclic radical having about nine to ten ring atoms in which at least one ring is aromatic. Specific and preferred values listed below for radicals, substituents, and ranges, are for illustration only; they do not exclude other defined values or other values within defined ranges for the radicals and substituents.
- the present invention encompasses any racemic, optically-active, polymorphic, or stereoisomeric form, or mixtures thereof, of a compound of the invention, which possess the useful properties described herein, it being well known in the art how to prepare optically active forms (for example, by resolution of the racemic form by recrystallization techniques, by synthesis from optically-active starting materials, by chiral synthesis, or by chromatographic separation using a chiral stationary phase) and how to determine antitumor activity or utility as an antitumor imaging agent using the standard tests described herein, or using other similar tests which are well known in the art.
- (C,-C 14 )alkyl can be methyl, ethyl, propyl, isopropyl, butyl, iso-butyl, sec-butyl, pentyl, 3-pentyl, hexyl, heptyl, octyl, nonyl, decyl undecyl, dodecyl, tridecyl or tetradecyl.
- (C 2 -C 14 )alkenyl can be vinyl, allyl, 1-propenyl, 2- propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1,-pentenyl, 2-pentenyl, 3-pentenyl, 4- pentenyl, 1- hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 1-heptenyl, 2- heptenyl, 3-heptenyl, 4-heptenyl, 5-heptenyl, 6-heptenyl, 1-octenyl, 2-octenyl, 3- octenyl, 4-octenyl, 5-octenyl, 6-octenyl, 7-octenyl, 1 -nonenyl, 2-nonenyl, 3- nonenyl, 4-nonenyl, 5-nonenyl, 6-nonenyl, 7-n-n
- (C 2 -C 14 )alkynyl can be ethynyl, 1-propynyl, 2- propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 1- hexynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, 5-hexynyl, 1-heptynyl, 2-heptynyl, 3-heptynyl, 4-heptynyl, 5-heptynyl, 6-heptynyl, 1-octynyl, 2- octynyl, 3-octynyl, 4-octynyl, 5-octynyl, 6-octynyl, 7-octynyl, 1-nonylyl,
- aryl can be phenyl, indenyl, or naphthyl.
- (C 3 -C 8 )cycloalkyl can be cyclopropyl, cyclobutyl, cyclcopentyl, cyclohexyl, cycloheptyl or cyclooctyl.
- amino acid is a natural amino acid residue (e.g. Ala, Arg, Asn, Asp, Cys, Glu, Gin, Gly, His, Hyl, Hyp, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, and Val) in D or L form, as well as unnatural amino acid (e.g.
- phosphoserine phosphothreonine; phosphotyrosine; hydroxyproline; gamma-carboxyglutamate; hippuric acid; octahydroindole-2-carboxylic acid; statine; l,2,3,4,-tetrahydroisoquinoline-3-carboxylic acid; penicillamine; ornithine; citruline; ⁇ -methyl-alanine; para-benzoylphenylalanine; phenylglycine; propargylglycine; sarcosine; and tert-butylglycine) residue having one or more open valences.
- the term also comprises natural and unnatural amino acids bearing conventional amino protecting groups (e.g.
- acetyl, acyl, trifluoroacetyl, or benzyloxycarbonyl as well as natural and unnatural amino acids protected at carboxy (e.g. as a (C,-C 6 )alkyl, phenyl or benzyl ester or amide) with conventional protecting groups.
- carboxy e.g. as a (C,-C 6 )alkyl, phenyl or benzyl ester or amide
- suitable amino and carboxy protecting groups are known to those skilled in the art (See for example, T.W. Greene, Protecting Groups In Organic Synthesis; Wiley: New York, 1981; D. Voet, Biochemistry, Wiley: New York, 1990; L. Stryer, Biochemistry. (3rd Ed.), W.H. Freeman and Co.: New York, 1975; J. March, Advanced Organic
- the amino or carboxy protecting group can also comprise a radionuclide (e.g., Fluorine-18, Iodine-123, or Iodine- 124).
- a radionuclide e.g., Fluorine-18, Iodine-123, or Iodine- 124.
- a "peptide” is a sequence of 2 to 25 amino acids (e.g. as defined hereinabove) or peptidic residues having one or more open valences.
- the sequence may be linear or cyclic.
- a cyclic peptide can be prepared or may result from the formation of disulfide bridges between two cysteine residues in a sequence.
- a peptide can be linked through the carboxy terminus, the amino terminus, or through any other convenient point of attachment, such as, for example, through the sulfur of a cysteine.
- Peptide derivatives can be prepared as disclosed in U.S. Patent Numbers 4,612,302; 4,853,371 ; and 4,684,620, or as described in the Examples hereinbelow. Peptide sequences specifically recited herein are written with the amino terminus on the left and the carboxy terminus on the right.
- the peptide can be poly-L-lysine, poly-L-glutamic acid, poly-L-aspartic acid, poly-L-histidine, poly-L-ornithine, poly-L-serine, poly-L-threonine, poly-L-tyrosine, poly-L-lysine— L-phenylalanine or poly-L- lysine-L-tyrosine.
- a "residue of a compound of formula I" is a radical of a compound of formula I having one or more open valences. Any synthetically feasible atom or atoms of the compound of formula I may be removed to provide the open valence, provided the resulting compound is able to localize in or near tumors. Based on the linkage that is desired, one skilled in the art can select suitably functionalized starting materials that can be derived from a compound of formula I using procedures that are known in the art.
- suitable atoms that may be removed include the NH 2 group of the a-carboxamide (illustrated in figure 1) or a hydrogen atom from the NH 2 group of the a- carboxamide, the NH 2 group of the b-carboxamide (illustrated in figure 1) or a hydrogen atom from the NH 2 group of the b-carboxamide, the NH 2 group of the d-carboxamide (illustrated in figure 1) or a hydrogen atom from the NH 2 group of the d-carboxamide, the NH 2 group of the e-carboxamide (illustrated in figure 1) or a hydrogen atom from the NH 2 group of the e-carboxamide, and X at the 6- position (illustrated in figure 1).
- the hydrogen atom of the hydroxy group at the 3' position of the sugar may be removed.
- adenosyl is an adenosine radical in which any synthetically feasible atom or group of atoms have been removed, thereby providing an open valence. Synthetically feasible atoms which may be removed include the hydrogen atom of the hydroxy group at the 5' position. Accordingly, adenosyl can conveniently be attached to the 6-position of a compound of formula I via the 5' position of adenosyl.
- a "molecule comprising B-10" can be any compound that contains at least one B-10 atom.
- the nature of the molecule that includes B-10 is not critical. The compound, however, must be nontoxic and must be able to enter the tumor cell or locate near the tumor cell when the molecule comprising B-10 is attached.
- a "residue of a molecule comprising B-10" is a radical of a molecule comprising B-10 having one or more open valences. Any synthetically feasible atom or atoms of the molecule comprising B-10 may be removed to provide the open valence, provided bioactivity is substantially retained. Based on the linkage that is desired, one skilled in the art can select suitably functionalized starting materials that can be derived from a molecule comprising B-10 using procedures that are known in the art.
- a "residue of o-carborane,” a “residue of m- carborane,” or a “residue of p-carborane” is a radical of o-carborane, m- carborane or p-carborane, respectively, having one or more open valences. Any synthetically feasible atom or atoms of o-carborane, m-carborane or p-carborane may be removed to provide the open valence, provided bioactivity is substantially retained.
- the invention provides a compound of formula I ( Figure 1) linked to one or more molecules comprising B-10, wherein X is CN, OH, CH 3 , adenosyl or a molecule comprising B-10; or a pharmaceutically acceptable salt thereof.
- a variety of molecules comprising B-10 known in the art are useful in the present invention.
- the molecules vary considerably in structure but are suitable to practice the present invention.
- Acceptable species include boron containing amino acids, carbohydrates, and nucleosides, as well as carboranes.
- a wide variety of boron-containing compounds are commercially available, or are known in the art.
- a variety of molecules comprising B-10 are commercially available from Boron Biologicals, Inc., Raleigh, North Carolina and RysCor Science, Inc., Raleigh, North Carolina.
- at least one molecule comprising B-10 can be o- carborane, m-carborane or p-carborane. More specifically, at least one molecule comprising B-10 is o-carborane.
- o-Carborane [l,2-dicarbadodecaborane(12)]; m-carborane [l,7-dicarbadodecaborane(12)]; and p-carborane [1,12- dicarbadodecaborane(12)] are commercially available from Aldrich, Milwaukee, WI.
- each molecule comprising B-10 can independently be o-carborane, m-carborane or p-carborane. More specifically, each molecule comprising B-10 is o-carborane.
- Such a linkage can be formed from suitably functionalised starting materials using synthetic procedures that are known in the art. Based on the linkage that is desired, one skilled in the art can select suitably functional starting materials that can be derived from a residue of a compound of formula I and from a given residue of a molecule comprising B-10 using procedures that are known in the art.
- the residue of the molecule comprising B-10 can be directly linked to any synthetically feasible position on the residue of a compound of formula I.
- Suitable points of attachment include, for example, the b- carboxamide, the d-carboxamide, and the e-carboxamide (illustrated in figure 1), as well as the 6-position (the position occupied by X in figure 1), and the 5'- hydroxy and the 3'-hydroxy groups on the 5-membered sugar ring, although other points of attachment are possible.
- 5,739,313 discloses compounds (e.g., cyanocobalamin-b-(4-aminobutyl)amide, methylcobalamin-b- (4-aminobutyl)amide, and adenosylcobalamin-b-(4-aminobutyl)amide) that are useful intermediates for the preparation of compounds of the present invention.
- Compounds wherein the residue of a molecule comprising B-10 is linked to the 6-position of a compound of formula I can be prepared by reducing a corresponding Co (II) compound of formula I to form a nucleophihc Co (I) compound and treating this Co (I) compound with a residue of a molecule comprising B-10 (or a derivative thereof) comprising a suitable leaving group, such as a halide (e.g., a chloride).
- a suitable leaving group such as a halide (e.g., a chloride).
- the invention also provides compounds having more than one residue of a molecule comprising B-10 directly linked to a compound of formula I.
- the residue of a molecule comprising B-10 can be directly linked to a residue of the b-carboxamide of the compound of formula I and a residue of another molecule comprising B-10 can be directly linked to a residue of the d- carboxamide of the compound of formula I.
- the residue of a molecule comprising B-10 can be directly linked to the 6-position of the compound of formula I and a residue of another molecule comprising B-10 can be directly linked to a residue of the d- or e-carboxamide of the compound of formula I.
- the residue of a molecule comprising B-10 can also be linked to the residue of a compound of formula I by a suitable linker.
- the structure of the linker is not crucial, provided it yields a compound of the invention which has an effective therapeutic index against the target cells, and which will localize in or near tumor molecules.
- Suitable linkers include linkers that separate the residue of a compound of formula I and the residue of a molecule comprising B-10 by about 5 angstroms to about 200 angstroms.
- Other suitable linkers include linkers that separate the residue of a compound of formula I and the residue of a molecule comprising B-10 by about 5 angstroms to about 100 angstroms, as well as linkers that separate the residue of a compound of formula I and the residue of a molecule comprising B-10 by about 5 angstroms to about 50 angstroms, or by about 5 angstroms to about 25 angstroms.
- Suitable linkers are disclosed, for example, in U.S. Patent No. 5,735,313.
- the linker can be linked to any synthetically feasible position on the residue of a compound of the residue of formula I.
- Suitable points of attachment include, for example, a residue of the b-carboxamide, a residue of the d-carboxamide, and a residue of the e-carboxamide, the 6-position (i.e., the position occupied by X in the compound of formula I), as well as a residue of the 5'-hydroxy group and a residue of the 3'-hydroxy group on the 5-membered sugar ring, although other points of attachment are possible.
- suitably functionalized starting materials that can be derived from a compound of formula I and from a given molecule comprising B-10 using procedures that are known in the art.
- the linkage can be formed from suitably functionalised starting materials using synthetic procedures that are known in the art. Based on the linkage that is desired, one skilled in the art can select suitably functional starting materials that can be derived from a residue of a compound of formula I, a residue of a molecule comprising B-10, and from a given linker using procedures that are known in the art.
- the linker can be a divalent radical of the formula W-
- the linker is a divalent radical, i.e., 1 , ⁇ -divalent radicals formed from a peptide or an amino acid.
- the peptide can comprise 2 to about 20 amino acids, 2 to about 15 amino acids, or 2 to about 12 amino acids.
- the peptide can be poly-L-lysine (i.e., [- NHCH[(CH 2 ) 4 NH 2 ]CO-] m -Q, wherein Q is H, (C,-C 14 )alkyl, or a suitable carboxy protecting group; and wherein m is about 2 to about 20.
- the poly-L-lysine can contains about 5 to about 15 residues (i.e., m is between about 5 and about 15). More specifically, the poly-L-lysine can contain about 8 to about 11 residues (i.e., m is between about 8 and about 11).
- the peptide can be poly-L-glutamic acid, poly-L- aspartic acid, poly-L-histidine, poly-L-ornithine, poly-L-serine, poly-L- threonine, poly-L-tyrosine, poly-L-lysine-L-phenylalanine or poly-L-lysine-L- tyrosine.
- the linker is prepared from 1 ,6-diaminohexane H 2 N(CH 2 ) 6 NH 2 , l,5-diaminopentane H 2 N(CH 2 ) 5 NH 2 , 1 ,4-diaminobutane H 2 N(CH 2 ) 4 NH 2 ., or 1,3-diaminopropane H 2 N(CH 2 ) 3 NH 2 .
- the linker can comprise one or more non-metallic radionuclides. Specifically, the linker can comprise more than one non-metallic radionuclides. More specifically, the linker can comprise 2 to about 10, 2 to about 8, 2 to about 6, or 2 to about 4 non-metallic radioisotopes.
- a specific residue of a peptide (i.e., linker) comprising one or more non-metallic radionuclides has the following formula wherein each M is independently a non- metallic radionuclide; each R is independently (C C 14 )alkyl, (C 2 - C 14 )alkenyl, (C 2 - [NHC Q
- R a and R b are each independently H or (C,-C 14 )alkyl; P; Q is H, (C r C 14 )alkyl, or a suitable carboxy protecting group; n is 2 to about 20; I is 1-5, j is 0-4 and I+j is ⁇ 5; or a pharmaceutically acceptable salt thereof.
- i can be 1, j can be 0, M can be Fluorine- 18, Bromine-76, or Iodine-123, and n can be about 6 to about 12.
- the molecule comprising B-10 can comprise one or more boron atoms.
- a suitable molecule comprising B-10 can contain 1 to about 20, 1 to about 15, 1 to about 10, or 1 to about 5 boron atoms.
- the molecule comprising B-10 can comprise one or more B-10 atoms.
- a suitable molecule comprising B-10 can contain 1 to about 20, 1 to about 15, 1 to about 10, or 1 to about 5 B-10 atoms.
- the molecule comprising B-10 can be an amino acid, a carbohydrate, a nucleoside or a carborane. Specifically, the molecule comprising B-10 is o-carborane, m-carborane or p-carborane.
- Compounds wherein the linker is linked to the 6-position of a compound of formula I can be prepared by preparing a nucleophihc Co (I) species as described herein above, and reacting it with a linker comprising a suitable leaving group, such as a halide (e.g. a chloride).
- the invention also provides compounds having more than one molecule comprising B-10 attached to a compound of formula I, each through a linker.
- the residue of a molecule comprising B-10 can conveniently be linked, through a linker, to a residue of the b-carboxamide of the compound of formula I and a residue of another molecule comprising B-10 can conveniently be linked, through a linker, to a residue of the d- or e-carboxamide of the compound of formula I.
- the residue of a molecule comprising B-10 can conveniently be linked, through a linker, to the 6-position of the compound of formula I and a residue of another molecule comprising B-10 can conveniently be linked, through a linker, to a residue of the b-, d- or e- carboxamide of the compound of formula I.
- the invention also provides compounds having more than one molecule comprising B-10 attached to a compound of formula I, either directly or through a linker.
- the residue of a molecule comprising B-10 can conveniently be linked, either directly or through a linker, to a residue of the b- carboxamide of the compound of formula I and a residue of another molecule comprising B-10 can conveniently be linked, either directly or through a linker, to a residue of the d- or e-carboxamide of the compound of formula I.
- the residue of a molecule comprising B-10 can conveniently be linked, either directly or through a linker, to the 6-position of the compound of formula I and a residue of another molecule comprising B-10 can conveniently be linked, either directly or through a linker, to a residue of the b-, d- or e-carboxamide of the compound of formula I.
- U.S. Patent Number 5,739,313 discloses cobalamin analogs comprising a compound of formula I, a linking group, and a chelating group comprising a detectable radionuclide or a paramagnetic ion.
- the compounds are disclosed to localize in tumor cells following administration, and to be useful for imaging tumors.
- the metallic radionuclide Gadolinium- 157 is an especially useful ion for conducting magnetic resonance imaging. It is also a useful target ion for neutron capture therapy.
- incorporation of Gd- 157 into one of the chelating molecules disclosed in U.S. Patent Number 5,739,313, provides a compound that is not only particularly useful for conducting magnetic resonance imaging, but also a compound that can be used in conjunction with neutron capture therapy, to treat tumors.
- each R is independently H or (C
- the invention also provides a compound wherein a residue of a compound of formula I ( Figure 1) is linked 1) to a residue of a molecule comprising B-10, and 2) to a detectable radoinuclide, wherein X is CN, OH, CH 3 , adenosyl, or a molecule comprising B-10; or a pharmaceutically acceptable salt thereof.
- the detectable radoinuclide can be directly linked to the residue of a compound of formula I, or can be linked by a linker to the residue of a compound of formula I.
- the linker can be any suitable linker described herein.
- the linker can also be a peptide or an amino acid.
- the invention also provides a compound wherein a residue of a compound of formula I (Figure 1) is linked 1) to a detectable radoinuclide; and 2) to a group comprising Gd-157; or a pharmaceutically acceptable salt thereof.
- the Gd-157 can be linked to the residue of a compound of formula I by any suitable means.
- the detectable radoinuclide can be directly linked to the residue of a compound of formula I, or can be linked by a linker to the residue of a compound of formula I.
- the linker can be any suitable linker described herein.
- any detectable non-metallic radionuclide that is suitable for imaging can be used in the compounds of the invention.
- suitable non-metallic radionuclides include Carbon- 11 , Fluorine- 18, Bromine-76, and Iodine-123.
- the non-metallic radionuclide can be a non-metallic paramagnetic atom (e.g., Fluorine- 19); or a non-metallic positron emitting radionuclide (e.g., Carbon-11, Fluorine-18, Iodine-123, or Bromine-76).
- Metallic Radionuclide Suitable metallic radionuclides include Antimony- 124, Antimony- 125, Arsenic-74, Barium-103, Barium-140, Beryllium-7, Bismuth-206, Bismuth-207, Cadmium- 109, Cadmium-115m, Calcium-45, Cerium-139, Cerium-141, Cerium-144, Cesium-137, Chromium-51, Cobalt-55, Cobalt-56, Cobalt-57, Cobalt-58, Cobalt-60, Cobalt-64, Copper-67, Erbium- 169, Europium- 152, Gallium-64, Gallium-68, Gadolinium- 153, Gadolinium- 157 Gold-195, Gold-199, Hafiiium- 175, Hafiiium- 175-181, Holmium-166, Indium-110, Indium-I l l, Iridium
- a "detectable radionuclide” is any suitable radionuclide (i.e., radioisotope) capable of detecting cancer or other neoplastic cells in a diagnostic procedure in vivo or in vitro.
- Suitable detectable radionuclides include metallic radionuclides (i.e., metallic radioisotopes) and non-metallic radionuclides (i.e., non-metallic radioisotopes).
- a "therapeutic radionuclide” is any suitable radionuclide (i.e., radioisotope) that possesses therapeutic efficacy against cancer or other neoplastic cells in vivo or in vitro.
- Suitable therapeutic radionuclides include metallic radionuclides (i.e., metallic radioisotopes). Chelating groups
- any suitable chelating group can be incorporated into the compounds of the invention.
- Suitable chelating groups include those disclosed in U.S. Patent Number 5,739,313.
- the chelating group can be NT A, HEDTA, DCTA, RP414, MDP, DOTATOC, CDTA, HYNIC, EDTA, DTP A, TETA, DOTA, DOTMP, DCTA, 15N4, 9N3, 12N3, or MAG3 (or another suitable polyamino acid chelator), which are described herein below, or a phosphonate chelator (e.g. EDMT).
- the chelating group can be DTP A.
- DTP A is diethylenetriaminepentaacetic acid
- TETA is 1,4,8,11- tetraazacyclotetradecane-N,N',N",N'"-tetraacetic acid
- DOTA is 1,4,7,10- tetraazacyclododecane-N,N',N",N"'-tetraacetic acid
- 15N4 is 1,4,8,12- tetraazacyclopentadecane-N,N',N",N'"-tetraacetic acid
- 9N3 is 1,4,7- 5 triazacyclononane-N,N',N"-triacetic acid
- 12N3 is 1,5,9-triazacyclododecane- N,N',N"-triacetic acid
- MAG3 is (N-[N-[N-[(benzoylthio) acetyl] glycyl]glycyl]glycine); and DCTA is a cycl
- R 3 may by (C t -C 4 )alkyl or CH 2 CO 2 -, which may be attached through positions 4 or 5, or through the group R 3 and which carries from 1 to 4 detectable metal or nonmetal cations (M), monovalent cations, or the alkaline earth metals.
- M metal or nonmetal cations
- each individual cyclohexane-based 0 molecule may carry up to 4 metal cations (where both R 3 groups are
- NT A, HEDTA, and DCTA are disclosed in Poster Sessions, 5 Proceedings of the 46th Annual Meeting, J. Nuc.Med.. p. 316, No. 1386.
- RP414 is disclosed in Scientific Papers, Proceedings of the 46th Annual Meeting, Nuc.Med., p. 123, No. 499.
- MDP is disclosed in Scientific Papers, Proceedings of the 46th Annual Meeting, J. Nuc.Med.. p. 102, No. 413.
- DOTATOC is disclosed in Scientific Papers, Proceedings of the 46th Annual Meeting, L 0 Nuc.Med., p. 102, No. 414 and Scientific Papers, Proceedings of the 46th Annual Meeting, J. Nuc.Med., p. 103, No. 415.
- CDTA is disclosed in Poster Sessions, Proceedings of the 46th Annual Meeting, J. Nuc.Med., p. 318, No. 1396.
- HYNIC is disclosed in Poster Sessions, Proceedings of the 46th Annual Meeting, J. Nuc.Med.. p. 319, No. 1398.
- Bifunctional chelators i.e., chelating groups
- macrocyclic ligands in which conjugation is via an activated arm attached to the carbon backbone of the ligand can also be employed as a chelating group, as described by M. Moi et al., J. Amer. Chem., Soc, 49, 2639 (1989) (2-p- nitrobenzyl-l,4,7,10-tetraazacyclododecane-N,N',N",N'"-tetraacetic acid); S. V. Deshpande et al., J. Nucl. Med., 31, 473 (1990); G. Kuser et al., Bioconj. Chem., 1, 345 (1990); C.
- diagnostic chelator or therapeutic chelating groups can be any of the chelating groups disclosed in Scientific Papers, Proceedings of the 46th Annual Meeting, J. Nuc. Med., Wednesday, June 9, 1999, p. 124, No. 500.
- a “detectable chelating group” is a chelating group comprising a metallic radionuclide (e.g., a metallic radioisotope) capable of detecting cancer or other neoplastic cells in vivo or in vitro.
- a metallic radionuclide e.g., a metallic radioisotope
- the chelating group can be any one of the carbonyl complexes disclosed in Waibel et al., Nature Biotechnology, 897-901, Vol. 17, September 1999; or Sattelberger et al., Nature Biotechnology. 849-850, Vol. 17, September 1999.
- the detectable chelating group can be any of the carbonyl complexes disclosed in Waibel et al., Nature Biotechnology, 897-901, Vol. 17, September 1999; or Sattelberger et al., Nature Biotechnology, 849-850, Vol. 17, September 1999, further comprising a metallic radionuclide. More specifically, the detectable chelating group can be any of the carbonyl complexes disclosed in Waibel et al., Nature Biotechnology, 897-901, Vol. 17, September 1999; or Sattelberger et al., Nature Biotechnology, 849-850, Vol. 17, September 1999, further comprising Technetium-99m.
- a "therapeutic chelating group” is a chelating group comprising a metallic radionuclide (e.g., a metallic radioisotope) that possesses therapeutic efficacy against cancer or other neoplastic cells in vivo or in vitro. Any suitable chelating group can be employed.
- the therapeutic chelating group can be any of the carbonyl complexes disclosed in Waibel et al., Nature Biotechnology, 897-901, Vol. 17, September 1999; or Sattelberger et al., Nature Biotechnology, 849-850, Vol. 17, September 1999, further comprising a metallic radionuclide. More specifically, the therapeutic chelating group can be any of the carbonyl complexes disclosed in Waibel et al., Nature Biotechnology. 897-901, Vol. 17, September 1999; or Sattelberger et al., Nature Biotechnology. 849-850, Vol. 17, September 1999, further comprising Rhenium-186 or Rhenium-188.
- Tumors treatable with the compounds and methods of the invention can be located in any part of the mammal.
- the tumor can be located in the breast, lung, thyroid, lymph node, genitourinary system (e.g., kidney, ureter, bladder, ovary, teste, or prostate), musculoskeletal system (e.g., bones, skeletal muscle, or bone marrow), gastrointestinal tract (e.g., stomach, esophagus, small bowel, colon, rectum, pancreas, liver, or smooth muscle), central or peripheral nervous system (e.g., brain, spinal cord, or nerves), head and neck tumors (e.g., ears, eyes, nasopharynx, oropharynx, or salivary glands), or the heart.
- genitourinary system e.g., kidney, ureter, bladder, ovary, teste, or prostate
- musculoskeletal system e.g., bones, skeletal muscle, or bone m
- the compounds disclosed herein can be prepared using procedures similar to those described in U.S. Patent Number 5,739,313, or using procedures similar to those described herein.
- the residue of a molecule comprising B-10 can be linked to the residue of a compound of formula I as described hereinabove. Additional intermediates and synthetic procedures useful for preparing compounds of the invention are disclosed, for example, in Hogenkamp, H. et al., Synthesis and Characterization of nido-Carborane- Cobalamin Conjugates, Nucl. Med.
- Examples of pharmaceutically acceptable salts are organic acid addition salts formed with acids which form a physiological acceptable anion, for example, tosylate, methanesulfonate, acetate, citrate, malonate, tartarate, succinate, benzoate, ascorbate, -ketoglutarate, and ⁇ -glycerophosphate.
- Suitable inorganic salts may also be formed, including hydrochloride, sulfate, nitrate, bicarbonate, and carbonate salts.
- compositions may be obtained using standard procedures well known in the art, for example by reacting a sufficiently basic compound such as an amine with a suitable acid affording a physiologically acceptable anion.
- Alkali metal (for example, sodium, potassium or lithium) or alkaline earth metal (for example calcium) salts of carboxylic acids can also be made.
- the compounds of the present invention can be formulated as pharmaceutical compositions and administered to a mammalian host, such as a human patient in a variety of forms adapted to the chosen route of administration, i.e., orally or parenterally (e.g., by intravenous, intramuscular, intraperitoneal). Preferably, the compounds are administered parenterally.
- the active compound may also be administered intravenously or intraperitoneally by infusion or injection.
- Solutions of the active compound or its salts can be prepared in water, optionally mixed with a nontoxic surfactant.
- Dispersions can also be prepared in glycerol, liquid polyethylene glycols, triacetin, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.
- the pharmaceutical dosage forms suitable for injection or infusion can include sterile aqueous solutions or dispersions or sterile powders comprising the active ingredient which are adapted for the extemporaneous preparation of sterile injectable or infusible solutions or dispersions, optionally encapsulated in liposomes.
- the ultimate dosage form should be sterile, fluid and stable under the conditions of manufacture and storage.
- the liquid carrier or vehicle can be a solvent or liquid dispersion medium comprising, for example, water, ethanol, a polyol (for example, glycerol, propylene glycol, liquid polyethylene glycols, and the like), vegetable oils, nontoxic glyceryl esters, and suitable mixtures thereof.
- the proper fluidity can be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersions or by the use of surfactants.
- the prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, buffers or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.
- Sterile injectable solutions are prepared by incorporating the active compound in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filter sterilization.
- the preferred methods of preparation are vacuum drying and the freeze drying techniques, which yield a powder of the active ingredient plus any additional desired ingredient present in the previously sterile-filtered solutions.
- suitable doses of a compound of the invention for use in therapy, in conjunction with neutron capture include doses in the range of from about 0.1 ⁇ g to about 100 ⁇ g, e.g., from about 0.5 ⁇ g to about 50 ⁇ g, or from about 0.5 ⁇ g to 15 ⁇ g per treatment.
- Suitable doses for use in imaging or for use in imaging and therapy include doses in the range of from about 0.1 mg to about 50 g, e.g., from about .5 mg to about 10 g, or from about 0.5 g to 2 g per treatment.
- the desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example, as two, three, four or more sub-doses per day.
- the sub-dose itself may be further divided, e.g., into a number of discrete loosely spaced administrations.
- the compounds are preferably dissolved or dispersed in a nontoxic liquid vehicle, such as physiological saline or a similar aqueous vehicle, to the desired concentration.
- a preselected therapeutic unit dose is then administered to the test animal or human patient, by oral administration or ingestion or by parenteral administration, as by intravenous or intraperitoneal infusion or injection, to attain the desired in vivo concentration.
- Doses useful for treating tumors can be derived, from those found to be effective to treat tumors in humans in vitro or in animal models, such as those described hereinbelow, or from dosages of other labeled vitamin B 12 molecules, previously employed in animal therapy.
- Figure 2 and Figure 3 each illustrate the four step synthesis used to prepare three cyanocobalamin-nidQ-carborane conjugates.
- o-Carborane carboxylic acid (2) was prepared by reacting o-carborane with n-butyllithium and carbondioxide in ether for approximately one hour at -78 °C. Treatment with thionyl chloride gave o-carborane carboxylic acid chloride (3), which was allowed to react with 1 ,4-butanediamine in pyridine to give the amide linked nido-carboranoyl(4-aminobutyl)amide (4).
- Compound (4) was linked to the b- monocarboxylic acid of cyanocobalamin, the d-monocarboxylic acid of cyanocobalamin, or both the b- and d-dicarboxylic acid of cyanocobalamin.
- hydroxybenzotriazole and l-ethyl-3-(3-dimethylaminopropyl) carbodiimide was added to facilitate the formation of the amide bond.
- the invention may be further illustrated by the following examples.
- a linear gradient was used from 5% B to 30% B over 10 minutes and was held at 30% B for 20 minutes before returning to initial conditions (mono-carborane synthesis).
- the di- carborane required a longer gradient using the same mobile phases; 5% B to 65% B over 25 minutes and held at 65% B for 15 minutes before returning to initial conditions.
- the separations were monitored by UN absorption at 214 NM. The flow was 1.0 ml minute and was split post-column allowing - 10 ul to flow into the mass spectrometer.
- Mass spectral data was collected using electrospray ionization in positive mode over a mass range of 300 to 2300 AMU at a dwell time of 0.3 ms/0.1 AMU.
- Synthetic samples were prepared at 5 or 10 mg/ml in pump A mobile phase and an aliquot injected onto the HPLC (1-5 ul). Retention times of the mono- and di -carborane products were determined to be 13.3 minutes and 16.2 minutes respectively.
- Purification of the b and d mono-carborane products was achieved by collecting fractions at the elution times for several injections. The collected fractions were combined and dried to a powder. A portion of the purified product was dissolved in methanol:H 2 O (1 :1) and reanalyzed by HPLC- MS to ascertain the purity.
- Results included the identification of the b and d carborane cyanocobalamin (CCC) analogs by LC-MS. Products identified were subsequently separated and purified by HPLC. The purified products were reanalyzed by LC-MS with no starting material detected. MS/MS data was also obtained on the b and d CCC analogs to provide further structural characterization. These purified products were then tested in the biological assays. The e-CCC analog and the di-CCC analog were also separated and identified by LC-MS, however, the preparations contained more reaction side products resulting in difficult purification.
- CCC carborane cyanocobalamin
- the starting material 4 was prepared as follows (see Figure 2): a. o-carborane carboxylic acid (2).
- a solution of o- carborane (1) (5.0 g, 34.7 mmol) in 500 ml dry ether in a 1 L round bottom flask was cooled to -78°C in a dry ice-acetone bath. The solution was flushed with argon and the flask sealed with a serum stopper.
- n-Butyllithium 24 mL, 1.6 M in hexanes
- o-Carborane carboxylic acid (2.0 g, 10.6 mmol), dried over P 2 O 5 , was dissolved in 30 ml thionylchloride and heated under reflux for 3 hours. The solution was cooled to room temperature, evaporated to dryness and dried over P 2 O 5 (2.05 g, 9.9 mmol, 93%) to provide (3), which was used without further purification.
- Nido-Carboranoyl(4-aminobutyl) amide (4). The acid chloride (3) was dissolved in 15 ml dry pyridine and 1 ,4-diaminobutane (1.12 g, 12.7 mmol) was added.
- 6-(3-aminoprop-l-yl)cobalamin 300 mg
- hydroxybenzotriazole 270 mg
- l-ethyl-3-(3-dimethylaminopropyl)carbodiimide 382 mg
- o- carborane monocarboxylic acid 300 mg
- the reaction mixture was concentrated to remove acetone and desalted via phenol extraction.
- the concentrated aqueous solution was crystallized from aqueous acetone to give the title compound.
- the intermediate 6-(3-aminoprop-l-yl)cobalamin was prepared as follows: a.
- 6-(3-aminoprop-l-yl)cobalamin 6-(3-aminoprop-l-yl)cobalamin.
- Cyano cobalamin 500 mg was dissolved in 100 ml deoxygenated water containing 10 mg CoCl 2 , and reduced with sodium borohydride to give the corresponding Co (I) compound.
- 3-chloropropyl amine hydrochloride 130 mg was dissolved in 5 ml deoxygenated ethanol was added. After one hour, at room temperature, the mixture was desalted via phenol extraction. The aminopropylcarbolamine was back extracted into water after the addition of 1 volume of acetone and 3 volumes of ether. The aqueous solution was concentrated and the desired 6-(3-aminoprop-l-yl)cobalamin crystallized from aqueous acetone (yield 510 mg).
- Example 3 DTPA-aminopropylcarbalamin (DAPC) Using a coupling procedure similar to that described in Example
- Example 4 In vitro biological activity of the carborane cyanocobalamin analogs. To assess the in vitro binding of the carborane cyanocobalamin
- Example 1 CCC
- Example 3 DTPA-aminopropylcobalamin
- DAPC DTPA-aminopropylcobalamin
- the analogs competitively blocked Co-57-cyanocobalamin from binding to the transcobalamin proteins. Therefore, the cpm of the modified UBBC assay was significantly lower than that of the clinical runs.
Abstract
Description
Claims
Priority Applications (8)
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KR1020017013054A KR20020032421A (en) | 1999-04-16 | 2000-04-15 | Cobalamin conjugates useful as antitumor agents |
US10/777,820 US7468432B2 (en) | 1999-04-16 | 2004-02-12 | Cobalamin conjugates useful as antitumor agents |
US12/266,763 US20090060837A1 (en) | 1999-04-16 | 2008-11-07 | Cobalamin conjugates useful as antitumor agents |
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WO2001028592A1 (en) * | 1999-10-15 | 2001-04-26 | Mayo Foundation For Medical Education And Research | Cobalamin conjugates useful as imaging agents and as antitumor agents |
WO2001028595A1 (en) * | 1999-10-15 | 2001-04-26 | Mayo Foundation For Medical Education And Research | Cobalamin conjugates useful as imaging and therapeutic agents |
WO2002042318A2 (en) * | 2000-10-25 | 2002-05-30 | Mayo Foundation For Medical Education And Research | Transcobalamin receptor binding conjugates for neutron capture therapy |
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EP1435973A1 (en) * | 2001-09-28 | 2004-07-14 | Mayo Foundation For Medical Education And Research | Coadministration of transport protein with conjugated cobalamin to deliver agents |
US6806363B1 (en) | 1999-04-16 | 2004-10-19 | Mayo Foundation For Medical Education & Research | Cobalamin conjugates useful as antitumor agents |
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JP2007515457A (en) * | 2003-12-22 | 2007-06-14 | ソリダゴ・アーゲー | Cobalamin derivatives effective for diagnosis and treatment of abnormal cell proliferation |
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US7591995B2 (en) | 1999-10-15 | 2009-09-22 | Mayo Foundation For Medical Education And Research | Cobalamin conjugates useful as imaging and therapeutic agents |
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US6838073B1 (en) | 1999-10-15 | 2005-01-04 | Mayo Foundation For Medical Education And Research | Cobalamin conjugates useful as imaging and therapeutic agents |
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EP1435973A4 (en) * | 2001-09-28 | 2007-05-02 | Mayo Foundation | Coadministration of transport protein with conjugated cobalamin to deliver agents |
WO2004056840A1 (en) * | 2002-12-20 | 2004-07-08 | Astrazeneca Ab | Novel compounds useful in boron neutron capture therapy |
US7232805B2 (en) | 2003-09-10 | 2007-06-19 | Inflabloc Pharmaceuticals, Inc. | Cobalamin conjugates for anti-tumor therapy |
JP2007515457A (en) * | 2003-12-22 | 2007-06-14 | ソリダゴ・アーゲー | Cobalamin derivatives effective for diagnosis and treatment of abnormal cell proliferation |
CN112442095A (en) * | 2020-12-01 | 2021-03-05 | 南京艾斯特医药科技有限公司 | Refining method of vitamin B12 and application of obtained product |
CN112442095B (en) * | 2020-12-01 | 2022-07-22 | 上海科黛生物科技有限公司 | Refining method of vitamin B12 and application of obtained product |
Also Published As
Publication number | Publication date |
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WO2000062808A8 (en) | 2001-12-06 |
KR20020032421A (en) | 2002-05-03 |
WO2000062808A3 (en) | 2001-02-22 |
CA2370419A1 (en) | 2000-10-26 |
AU4243700A (en) | 2000-11-02 |
EP1171162A2 (en) | 2002-01-16 |
MXPA01010395A (en) | 2004-08-12 |
JP2002542207A (en) | 2002-12-10 |
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