JP2002542207A - Cobalamin conjugates useful as antitumor agents - Google Patents

Abstract

The present invention provides a cobalamin compound that binds to a neutron capture therapeutic target (eg, boron-10 or gadolinium-157) and optionally binds to a detectable moiety, and further comprises a pharmaceutical composition comprising the compound. Also provided are methods of using the compounds in medical diagnosis and therapy.

Description

DETAILED DESCRIPTION OF THE INVENTION

RELATED APPLICATIONS [0001] This application is related to US Provisional Patent Application No. 60 / 129,333, filed April 16, 1999; and US Provisional Patent Application No. 60, filed October 15, 1999. / 159,873 claim priority.

BACKGROUND OF THE INVENTION Boron neutron capture therapy involves irradiating a stable isotope ¹⁰ B with low energy (0.025 eV) or thermal neutrons to generate helium nuclei (alpha particles) and ⁷ Li nuclei. Based on nuclear reactions.

[0003] The therapeutic potential of this response was recognized by Locher over 50 years ago (Locher, GL et al., Am. J. Roentgenol Radium).
Ther. , 36, 1-13 (1936)), boron neutron capture therapy (B
Sweet was the first to suggest that NCT was useful in treating brain tumors (Javid, M. et al., J. Clin. Invest, 31, 603-).
610 (1952); Sweet, W.C. H. , N .; Engl. J. Med. , 2
45, 875-878 (1951); Sweet, W.C. H et al. Neuro
osurg, 9, 200-209 (1952)).

Shortly thereafter, Brookhaven National Laboratory, in cooperation with Sweet et al., Launched a clinical trial using borax as a capture agent at Massachusetts General Hospital (
Farr, L .; E. FIG. Et al., Am. J. Roentgenol, 71, 279-2
91 (1954); Godwin, J .; T. Other, Cancer (Phila.)
8, 601-615 (1995)). The goal at that time was to use BNCT as a surgical adjunct to treat patients with the most malignant and difficult-to-treat brain tumor, glioblastoma multiforme.

[0005] Further clinical trials were performed in the early 1960's, but these showed no evidence of therapeutic efficacy (Farr, LE et al., Supra; Godwin, JT. Et al., Supra; Asbury, A. K. et al., J. Neupatholol. Exp.
urol. , 31, 278-303 (1972)), with side effects in normal tissues (Asbury, AK, et al., Supra). More promising clinical studies for the treatment of malignant gliomas by Hatanaka et al. And the melanomas of Misima et al. (Mishima, Y. et al., Lancet, 2, 388-289 (19)
89)) (Hatanaka, HA, J. Neurol.,
209, 81-94 (1975); Hatanaka, H .; Other, Boro
n Neutron Capture Therapy for Tumors
, Chap. 25, pp. 349-378, Niigata; Nishimura Co., Ltd. in Japan), and interest in BNCT in Japan and overseas has been revived.

[0006] Advantages of the theoretical BNCT is that a two-component or binary system consisting of ^{10 B} and thermal neutrons, the ^{10 B} and thermal neutrons bind, without causing significant damage to normal tissue, tumor cells Produces high linear energy imparting (LET) radiation that can selectively destroy For a successful BNCT, a critical amount of ¹⁰ B and a sufficient number of thermal neutrons need to be delivered to individual tumor cells.

[0007] Over the past few years, the Department of Energy and the NIH have renewed their financing of BNCT-related research, which has supported an increasing number of researchers in many different fields. Advances in BNCT in the areas of compound distribution and pharmacokinetics compare favorably with other emerging modalities, such as the use of radiolabeled antibodies for cancer therapy using photon activation therapy, photodynamic therapy, and physiological targeting There is no.

There are many nuclides that have a high tendency to absorb low energy or thermal neutrons, and this property, called neutron capture cross section (σ), is measured by burn (1b = 10 ⁻²⁴ c
m ² ). Of the various nuclides with a large neutron capture cross section, ¹⁰ B is most attractive for the following reasons: (a) it is non-radioactive, readily available, and contains about 20% natural boron: (b) capture Particles released by the reaction [ ¹⁰ B (n, α) ⁷ Li
] Is a very high LET: (c) its path length is about 1 cell diameter (10-14
μm) and the radiation effect is limited to those tumor cells that have absorbed a theoretically sufficient amount of ¹⁰ B, while at the same time not harming normal cells: and (d) due to the additional boron chemistry, It can be contained in many different chemical structures.

[0009] ⁷ Li and α particles are the primary fission products of the neutron capture reaction with ¹⁰ B. Alpha particles are relatively slow, causing closely spaced ionization events consisting of well-defined column trajectories. These have a path length of about 10 μm, high LET, and destroy various bioactive molecules including DNA, RNA and proteins. For these reasons, little, if any, cell repair from damage due to α-particle induced radiation.

[0010] The ¹⁰ B (n, α) ⁷ Li reaction can only be performed in large amounts of biology in the presence of thermal neutrons localized around the cell, on the cell or inside the cell, and a sufficient amount of a critical amount of ¹⁰ B. The resulting radiation can be extremely localized in order to produce a positive effect and does not harm normal cellular components. Therefore, selectivity is both an advantage and a disadvantage of BCNT because tumor cells need to deliver boron 10 in larger quantities than normal cells.

[0011] The boron compound used in BNCT has high specificity for malignant cells and ideally has low concentrations in nearby normal tissues and blood. Because it is desirable to limit radiation to only these cells, intracellular and optimal nuclear localization of boron is preferred.

A number of boron-containing derivatives of chlorpromazine have been synthesized (Nakagw
a, T .; Chem. Pharm. Bull. (Tokyo), 24,
778-781 (1976); Alam, F .; Other, Sthralenthe
r Onkol, 165, 121-125 (1989)), for its in vivo tumor localization properties. p-Boronophenylalanine is another compound that has been studied as a potential capture agent in the treatment of melanoma. The principle of its use is its affinity for melanoma aromatic amino acids and its subsequent incorporation into melanin (Ichihashi, M. et al., J. Invest Der.
matol. 78, 215-218 (1982); Others, Neutral Capture Therapy, 230-236, Niigata, Japan: Nishimura Corporation (1986)).

[0013] Tumor localization after iv administration was determined by whole body autoradiography (Code
rre, J. et al. A. Other, Cancer Res. , 48, 6313-631
6 (1988)) in several patients with cutaneous melanoma after perilesional injection (M
ishima, Y .; Other, Sthralenther. Onkol. 165
, 251-254 (1989)). Numerous other boron-containing amino acids have been synthesized (Hall, IH, et al., Which, stimulated by the experience of Mishima, can potentially incorporate large amounts into proteins of malignant cells).
J. Pharm. Sci. , 68, 685-688 (1979)).

Another method of selective targeting of boron to melanoma is based on the observation that thiouracil is preferentially included in melanoma during melanogenesis (Whitt
acker, J .; R. , J. et al. Biol. Chem. , 246, 6217-
6226 (1971)). This observation highlights the synthesis of multiple boron-containing thiouracils (Gabel, D., Clinical Aspects).
of Neutron Capture Therapy, 233-241
, New York: Plenum Publishing Co. (1989)
), These are currently being evaluated in animals.

[0015] Two other compounds that have a propensity to localize in malignancies are porphyrins and related phthalocyanines. Although the biochemical principle for reaching high concentrations of these compounds in malignant tumors is unknown, this observation provides a rationale for the use of hematoporphyrin in cancer photochemotherapy (Daugherty.
T. J. Others, Porphyrin Photosensitization,
3-13, New York: Plenum Publishing Co. (
1981)).

Due to the high concentration of these compounds in tumors and their localization and persistence in cells, several groups of researchers have found that borated porphyrins (Kahl, SB et al., Neutron Capture Therapy, 61). −67, Niigata,
Japan: Nishimura Corporation (1986)) and phthalocyanine (Alam, F
. Others, Sthralenther Oncol, 165, 121-123
(1989) was synthesized as a potential capture agent. Borated porphyrin is N
than a ₂ B ₁₂ H ₁₁ monomers or dimers shaped SH, 3 to 4 times per unit dose in the cell culture appears to be effective (Laster, B
. H. Other, Sthralenther Oncol, 165, 203-2
05 (1989)). Liver concentrations of these compounds are also high (Kahl, SB).
. Et al., Supra), but this does not limit its use in capture agents for treating brain tumors.

The last class of low molecular weight boron compounds are boron-containing purines and pyrimidines, and their nucleosides. The rationale for developing them is that such compounds are selectively included in rapidly growing tumor cells and are trapped inside cells after being converted to the corresponding nucleotides. Alternatively, these bases and their nucleosides may function as analogs of the natural precursor of the nucleic acid and become incorporated into nuclear DNA.

[0018] These heavy particles resulting from the capture reaction deliver large amounts of energy to the nuclear target, which allows for lower boron concentrations than would be required if the compound were located extracellularly, The cytoplasmic localization, or preferably the nuclear localization, of all the boron compounds is advantageous (Gabel, D. et al., Radiat Re.
s, 111, 14-25 (1987); Fairchild, R .; G. FIG. Int. J. Radiat. Oncol. Biol. Phys. , 1
1, 831-840 (1985)). Schinazi and Prusoff (S
chinazi, R .; F. Et al., Tetrahedron. Lett. , 5
0,4981-4984 (1978)) synthesizes the first boron-containing nucleoside, an analog of thymidine, 5-dihydroxyboryl-2′-deoxyuridine, at a concentration of 1600 μM against African green monkey (Vero) cells. Levels were not cytotoxic (Laster, BH et al.
Neutron Capture Therapy, 46-54, Niigata, Japan: Nishimura Corporation (1986)). In vitro neutron emission studies of cells growing in the presence of 5-dihydroxy-2'-deoxyuridine indicate that a biological equivalent to ¹⁰ B at concentrations up to 6 μg is sufficient, if achievable in vivo, for BNCT. The effect occurred.

In the 1960's and early 1970's, there was an interest in the potential use of polyclonal antibodies targeting tumor-associated antigens to deliver drugs and radioisotopes to tumors (Pressman, D. et al., Cancer). Res.,
40, 3001-3007 (1957); Ghose, T .; Other, Br.
Med. J. , 1, 90-93 (1967); Ghose, T .; Other, Ca
ncer (Phila.), 29, 1398-1400 (1972)). 1
Soloway in 964 years, antibodies were suggested for the selective targeting of ¹⁰ B against tumors (Soloway, A.H.., Supra). Hawth
Orne et al., (Hawthorne, MF et al., J. Medical C.
hem. , 15, 449-452 (1972)) are antibodies of diazonium salts with 1- (4-aminophenyl) -1,2-dicarbo-closo-dodecarborane targeting bovine serum albumin and human and mouse histocompatibility. Inclusion in polyclonal antibodies targeting the antigen was reported (Hawtho).
rn, M .; F. Others, ibid.).

From in vitro experiments, as evidenced by reduced viability after neutron irradiation,
These immunoconjugates show that human as well as murine lymphocytes have the lethal ¹⁰ B (n, α
) 7 It was claimed that sufficient boron could be delivered to maintain the reaction. But,
The immunoconjugate contained only 0.2% by weight of natural boron, which corresponded to 6 atoms of ¹⁰ B / molecule of the antibody. Instead, there must have been another explanation for the observed decrease in cell viability. Sneath et al. (Sneath
, R .; L. Jr., Jr. , J. et al. Medicinal Chem. , 17, 7
96-799 (1974)) showed that water-solubilizing groups must be included in protein-bound borane to maintain protein solubility in aqueous systems.

Thereafter, the groups of the polyhedral borane derivative containing the protein-binding functional group were bound to the IgG molecule by the carbodiimide reaction without showing any signs of precipitation (Sneath,
R. L. Jr., Jr. , J. et al. Medicinal Chem. , 19, 12
90-1294 (1976)).

The last class of macromolecular species that is considered useful for the delivery of ¹⁰ B is what is termed “encapsulated complexes” such as liposomes, microspheres and low-density lipoproteins (Kahl, SB et al. Strahlenther Onkol.,
165, 137-139 (1989)). Theoretically, a large amount of ¹⁰ B could be encapsulated and these encapsulated complexes could be used to bind tumors to monoclonal antibodies using existing methods or by targeting endogenous expressed cell surface receptors. When targeted, these become powerful delivery systems. Again, due to the preferential localization of the reticuloendothelial system, methods need to be developed to minimize this and maximize tumor resorption.

Cobalamin Vitamin B₁₂Years after its isolation as cyanocobalamin in 1948
Meanwhile, cyanocobalamin and possibly its photolysis product hydroxocobalamin
Was estimated to be produced in humans. Since then, cyanocobalamin has been a vitamin B ₁₂ (Methylcobalamin and hydroxocobalamin) are the natural sources of this vitamin
It is known to be embryogenic.

The structures of these various forms are shown in FIG. 1, where X is CN, OH, C
H is ₃ or adenosyl. In the following, the term cobalamin refers to all molecules except the X group. The basic ring system without cobalt (Co) or side chains is called choline, and octadehydrocholine is called cholor. Figure 1 shows Merck Inde
x, Merck & Co. (11th edition, 1989)
Is above the plane defined by the choline ring and nucleotides are below the plane of the ring. The choline ring has six amidoalkyl (2, 3, 7, 8, 13 and 18 positions)
H ₂ NC (O) Alk) substituents are attached and are called ae and g, respectively. D. L. Anton et al. Amer. Chem. Soc. ,
102, 2215 (1980).

Methylcobalamin acts as a cytoplasmic coenzyme of 5N-methyltetrahydrofolate: homocysteine methyltransferase (methionine synthase, EC 2.1.1.13), catalyzing the production of methionine from homocysteine. Adenosylcobalamin is a mitochondrial coenzyme of methylmalonyl-CoA mutase (EC 5.4.99.2) and interconverts methylmalonyl-CoA and succinyl-CoA.

[0026] All forms of vitamin B ₁₂ (adenosyl -, cyano -, hydroxo - or methylcobalamin), to the biological activity, transport proteins, it is necessary to bind to the (intrinsic factor and transcobalamin II) . In particular gastrointestinal absorption of vitamin B _12, among factors that bound to the intrinsic factor receptors in the terminal end ileum - it depends on vitamin B ₁₂ complex. Similarly, absorption by intravascular transport and subsequent cell of vitamin B ₁₂ throughout the body is transcobalamin II and the cell membrane transcobalamin II
It depends on each receptor. After trans cobalt amine II- vitamin B ₁₂ complex has been absorbed, transport protein undergoes lysozyme degradation, thereby vitamin B ₁₂ is released into the cytoplasm. Then vitamin B ₁₂ in all forms, adenosyl by cell request -, hydroxo - are mutually converted into methylcobalamin. For example, A. E. FIG. Finkler et al., Arch Biochem. Bioph
ys, 120, 79 (1967); Hall et al. Cell. Ph
ysiol. , 133, 187 (1987); E. FIG. Rappazzo
Et al. Clin. Invest. , 51, 1915 (1972) and R
. See Soda et al., Blood, 65, 795 (1985).

[0027] Rapidly growing cells have been shown to have increased thymidine and methionine absorption (eg, ME van Eijkeren et al., Act.
a. Oncology. , 31, 539 (1992); Kob
ota et al. Nucl. Med. , 32, 2118 (1991); Higashi et al. Nucl. Med. , 34, 773 (199
3)). Methylcobalamin, like cobalt 57-cyanocobalamin, has increased absorption in rapidly dividing tissues, as methylcobalamin is directly involved in methionine synthesis and indirectly involved in thymidylic acid and DNA synthesis. (Eg, BA Cooper et al., Nature, 19).
1, 393 (1961); Flodh, Acta Radiol. S
uppl. , 284, 55 (1986); Bloomquist and others,
Experientia, 25, 294 (1969)). Furthermore, in several malignant cells, an upregulation of the number of transcobalmin II receptors has been shown during its accelerated thymidine inclusion and DNA synthesis (J.
Lindemans et al., Exp. Cell. Res. , 184, 44
9 (1989); Amagasaki et al., Blood, 26, 13
8 (1990) and J.M. A. Begley et al. Cell Physio
l. 156, 43 (1993)).

[0028] Vitamin B ₁₂ has a potentially more characteristics that can be an attractive in vivo tumor therapy agents. Vitamin B ₁₂ is water soluble, no known toxic, it is excessively excreted by glomerular filtration. Moreover, absorption of vitamin _{B 12} by administration of nitrous oxide and other drugs, potentially operation can (D. Swanson other, P
hamaceuticals in Medical Imaging, Mac
Millan Pub. Co. , NY (1990) pp. 621-628
).

[0029] ¹²⁵ I- vitamin _{B 12} derivative process of preparation are described in U.S. Patent No. 3,981,863, issued to Niswender et al. In this process, vitamin B ₁₂ is first subjected to mild hydrolysis, Houts mostly below (e) - discloses that including isomers, to produce a mixture of monocarboxylic acids. The mixture is reacted with p- (aminoalkyl) phenol to introduce a B ₁₂ acid in (one of the reaction by the free carboxyl groups) phenol group. Mixed substituent B ₁₂ derivatives are then treatment with iodine in the phenolic group substituents. This patent, such mixing ^{125 I-B} ₁₂ derivative that is produced used the prepared antibody relative to the mixture, are useful in the radioimmunoassay of B _12.

T. M. U.S. Pat. 4,465,775
Reported that components of the radiolabeled mixture of Miswender et al. Did not bind to IF with the same affinity. Houts is radioiodinated derivatives of the pure monocarboxylic (d) isomer, it was disclosed to be useful in assays B ₁₂ to use the IF. However, Houts generally monocarboxylic (d) isomers are labeled with fluorophores or enzymes, may be used in a competition assay of vitamin B ₁₂ in a liquid, suitable for the imaging and treatment of tumors and organs labeled of demand for vitamin B ₁₂ derivative is subsequently disclose that are present.

US Pat. No. 5,739,3 issued to Collins and Hogenkamp
No. 13 reported that a cobalamin analog containing a linking group and a chelating group optionally containing a detectable radionuclide or paramagnetic ion localizes to tumor cells and is useful for tumor imaging. .

[0032] Despite previous efforts to identify neutron capture agents that are localized at high concentrations within tumor cells and are useful in the treatment of cancer, neutron capture currently is useful in treating tumors. There is demand.

DISCLOSURE OF THE INVENTION The present invention is a residue of a compound of formula I attached to a residue of a molecule comprising boron-10 (ie, B-10), wherein X is CN, OH, CH ₃ , a compound of the invention which is a molecule comprising adenosyl or B-10; or a pharmaceutically acceptable salt thereof.

The present invention relates to compounds wherein the residue of the compound of formula I (FIG. 1) is linked to a group of formula QLW-Det, wherein X is CN, OH, CH ₃ , adenosyl or B-10. Or QLW-Det, wherein Det is a chelating group comprising Gd-157, L is or is not a linker, and W and Q are each independently
-N (R) C (= O)-, -C (= O) N (R)-, -OC (= O)-, -C (
= O) O-, -O-, -S-, -S (O)-, -S (O) _2- , -C (= O)-
, —N (R) — or a direct bond, wherein each R is independently H or (C ₁ -C ₆ )
An alkyl or a pharmaceutically acceptable salt thereof is provided.

The present invention relates to compounds wherein the residue of the compound of formula I (FIG. 1) is linked to the residue of a molecule comprising B-10, wherein the residue of the compound of formula I is of the formula QL- even under the W-Det is bonded, wherein X is CN, OH, CH _3, adenosyl, formula Q-L-W-Det
Or a molecule comprising B-10, Det is a chelating group comprising a therapeutic or diagnostic radionuclide, L is or is not a linker, and Q and W are each independently- N (R) C (= O)-, -C (= O) N (R)-,
-OC (= O)-, -C (= O) O-, -O-, -S-, -S (O)-, -S (
O) ₂ —, —C (＝ O) —, —N (R) — or a direct bond, wherein each R is independently H or (C ₁ -C ₆ ) alkyl, or pharmaceutically acceptable A salt thereof is also provided.

The present invention relates to compounds wherein the residue of the compound of formula I (FIG. 1) is linked to the residue of a molecule comprising B-10, wherein the residue of the compound of formula I is of the formula QL- X is also CN, OH, CH ₃ , adenosyl, a group of formula QLW-Det,
Or a molecule containing B-10, Det is a chelating group containing Gd-157, L is or is not a linker, and W and Q are each independently -N (R
) C (= O)-, -C (= O) N (R)-, -OC (= O)-, -C (= O) O
-, -O-, -S-, -S (O)-, -S (O) _2- , -C (= O)-, -N (
R) - or a direct bond, each R is independently H or (C 1 _{-C 6)} alkyl, or also provides a pharmaceutically acceptable salt thereof.

The present invention provides that the residue of the compound of formula I (FIG. 1) is bound to a detectable radionuclide and also to the residue of a molecule containing boron-10 (ie, B-10). (here X CN, OH, CH _3, adenosyl, or a residue of a molecule comprising a B-10) compounds have, or provides a pharmaceutically acceptable salt thereof.

The present invention relates to compounds wherein the residue of the compound of formula I (FIG. 1) is bound to a detectable radionuclide and is also bound to a group comprising Gd-157, wherein X is CN , OH,
CH _3, adenosyl, molecules including B-10, or a group containing Gd-157), or also provides a pharmaceutically acceptable salt thereof. In particular, the formula for the group containing Gd-157 may have a QLW-Det, where Det is a chelating group containing Gd-157, L is or is not a linker, and W and Q is each independently -N (R) C (= O)-, -C (= O) N (R)-, -OC (= O)-, -C
(= O) O-, -O-, -S-, -S (O)-, -S (O) _2- , -C (= O)
—, —N (R) — or a direct bond, wherein each R is independently H or (C ₁ -C ₆ ) alkyl.

The present invention relates to a compound of formula I (FIG. 1) wherein the residue is 1) a molecule comprising B-10 or Gd
-2) a compound bound to a chelating group containing -157 and bound to one or more residues of the formula QLW-Det, wherein each Det independently comprises a metal radionuclide Chelating groups; each L is independently a linker or not;
Are each independently -N (R) C (= O)-, -C (= O) N (R)-, -OC
(= O)-, -C (= O) O-, -O-, -S-, -S (O)-, -S (O) _2- , -C (= O)-, -N (R ) - or a direct bond, each R is independently H or (C 1 _{-C 6)} alkyl, or also provides a pharmaceutically acceptable salt thereof.

The present invention also provides a pharmaceutical composition comprising a compound of the present invention and a pharmaceutically acceptable carrier.

The present 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 performing neutron capture therapy. .

The invention comprises administering to a mammal a detectable amount of a compound of the invention in combination with a pharmaceutically acceptable vehicle effective to image a tumor, and detecting the presence of the compound. Also provided is a method of imaging a tumor in a mammal in need of such imaging, comprising:

The present invention also provides a compound of the present invention for use in medical treatment or diagnosis.

The present invention also provides the use of a compound of the present invention for the manufacture of a medicament for tumor imaging in a mammal (eg, a human).

The present 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 (eg, a human).

The present invention also provides intermediates disclosed in the present invention that are useful for preparing compounds of the present invention, as well as synthetic methods useful for preparing compounds of the present invention.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 shows the structure of cobalamin, where X is CN (cyano), OH, CH ₃ or adenosyl.

FIG. 2 shows the synthesis of cyanocobalamin-nido-carborane conjugate.

FIG. 3 shows the synthesis of a representative compound of the invention.

DETAILED DESCRIPTION OF THE INVENTION Applicants have discovered that certain cobalamin analogs are useful for treating tumors in combination with neutron capture therapy. Applicants have also discovered that certain cobalamin analogs are useful as neutron capture agents and tumor imaging agents.

The following definitions were used, unless otherwise stated. Halo is fluoro, chloro, bromo, or iodo. Alkyl, alkoxy, alkenyl, alkynyl and the like represent both linear or branched groups. However, references to individual radicals such as "propyl" include only straight chain radicals, and branched isomers such as "isopropyl" are specifically mentioned. Allyl represents a phenyl radical or an ortho-conjugated bicyclic carbocyclic radical having about 9 to 10 ring atoms in which at least one ring is aromatic.

The specific values or preferred values listed below for radicals, substituents and ranges are for illustration only and exclude other defined values of the radicals or substituents or other values within the defined ranges. do not do.

It will be appreciated by those skilled in the art that compounds of the present invention having a chiral center will recognize and exist in optically active and racemic forms. Some compounds may exhibit polymorphism. The present invention includes racemic, photoactive, polymorphic, stereoisomeric or mixtures thereof of the compounds of the present invention with the useful properties described herein, and methods for preparing photoactive forms ( For example, by resolving racemic system by recrystallization technique,
By synthesis from optically active starting materials, by chiral synthesis, or by chromatographic separation using a chiral stationary phase) and using the standard tests described herein or other similar tests known to those skilled in the art. It should be understood that the methods used to determine antitumor activity or utility as antitumor imaging agents are known to those of skill in the art.

In particular, (C ₁ -C ₁₄ ) alkyl is methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, pentyl, 3-pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, It can be dodecyl, tridecyl or tetrasil.

In particular, (C ₂ -C ₁₄ ) alkyl is 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-nonenyl, 8-nonenyl, 1-decenyl, 2- Decenyl, 3-decenyl, 4-decenyl,
5-decenyl, 6-decenyl, 7-decenyl, 8-decenyl, 9-decenyl, 1
-Undecenyl, 2-undecenyl, 3-undecenyl, 4-undecenyl, 5
-Undecenyl, 6-undecenyl, 7-undecenyl, 8-undecenyl, 9
-Undecenyl, 10-undecenyl, 1-dodecenyl, 2-dodecenyl, 3-
Dodecenyl, 4-dodecenyl, 5-dodecenyl, 6-dodecenyl, 7-dodecenyl, 8-dodecenyl, 9-dodecenyl, 10-dodecenyl, 11-dodecenyl,
1-tridecenyl, 2-tridecenyl, 3-tridecenyl, 4-tridecenyl,
5-tridecenyl, 6-tridecenyl, 7-tridecenyl, 8-tridecenyl, 9-tridecenyl, 10-tridecenyl, 11-tridecenyl, 12-tridecenyl, 1-tetradecenyl, 2-tetradecenyl, 3-tetradecenyl, 4-
Tetradecenyl, 5-tetradecenyl, 6-tetradecenyl, 8-tetradecenyl, 9-tetradecenyl, 10-tetradecenyl, 11-tetradecenyl, 12
-Tetradecenyl, 13-tetradecenyl.

In particular, (C ₂ -C ₁₄ ) alkyl is 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, 2-nonylyl, 3
-Nonylyl, 4-nonylyl, 5-nonylyl, 6-nonylyl, 7-nonylyl, 8-
Nonylyl, 1-decynyl, 2-decynyl, 3-decynyl, 4-decynyl, 5-decynyl, 6-decynyl, 7-decynyl, 8-decynyl, 9-decynyl, 1-undecynyl, 2-undecynyl, 3-undecynyl, 4-undecynyl, 5-undecynyl, 6-undecynyl, 7-undecynyl, 8-undecynyl, 9-undecynyl, 10-undecynyl, 1-dodecynyl, 2-dodecynyl, 3-dodecynyl, 4-dodecynyl, 5-dodecynyl, 6- Dodecinyl, 7-dodecinyl, 8
-Dodecinyl, 9-dodecinyl, 10-dodecinyl, 11-dodecinyl, 1-tridecinyl, 2-tridecinyl, 3-tridecinyl, 4-tridecinyl, 5-tridecinyl, 6-tridecinyl, 7-tridecinyl, 8-tridecinyl, 9-tridecinyl , 10-tridecinyl, 11-tridecynyl, 12-tridecynyl,
1-tetradecynyl, 2-tetradecynyl, 3-tetradecynyl, 4-tetradecynyl, 5-tetradecynyl, 6-tetradecynyl, 7-tetradecynyl, 8-
It can be tetradecynyl, 9-tetradecynyl, 10-tetradecynyl, 11-tetradecynyl, 12-tetradecynyl, 13-tetradecynyl.

In particular, “allyl” can be phenyl, indenyl or naphthyl.

In particular, (C ₃ -C ₈ ) cycloalkyl is cyclopropyl, cyclobutyl,
It can be cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl.

As used herein, “amino acid” refers to naturally occurring amino acid residues in D or L form (eg, Ala, Arg, Asn, Asp, Cys, Glu, Gln, Gly).
, His, Hyl, Hyp, Ile, Leu, Lys, Met, Phe, Pro
, Ser, Thr, Trp, Tyr, Val) as well as unnatural amino acids having one or more open valences (eg, phosphoserine, phosphothreonine; phosphotyrosine; hydroxyproline; γ-carboxyglutamic acid; hippuric acid; Hydroindole-2-carboxylic acid; statin; 1,2,3
4-tetrahydroisoquinoline-3-carboxylic acid; penicillamine; ornithine: α-methyl-alanine; p-benzoylphenylalanine; phenylglycine; propargylglycine; sarcosine; t-butylglycine) residue.
This term refers to natural and unnatural amino acids having conventional amino acid protecting groups (eg, acetyl, acyl, trifluoroacetyl or benzyloxycarbonyl) as well as carboxyl (eg, (C ₁ -C ₆ ) alkyl, phenyl or benzyl esters or (As amides) as well as natural and unnatural amino acids protected by conventional protecting groups. Other suitable amino and carboxyl protecting groups are
It is known to those skilled in the art (for example, TW Greene, Protect).
ing Group In Organic Synthesis; Wiley
: New York, 1981; Voet, Biochemistry, Wi
ley: New York, 1990; Stryer, Biochemist
ry (3rd edition); H. Freeman and Co. : New York, 1
975; March, Advanced Organic Chemis
try Reactions Mechanisms and Structu
re (2nd edition), McGraw Hill: Knee York, 1977; Ca
rey and R.S. Sundberg, Advanced Organic
Chemistry Part B Reactions and Synt
hesis (second edition), Plenum: New York, 1977; and references cited herein). According to the present invention, the amino or carboxy protecting group is
It may also contain radionuclides (eg, fluorine-18, iodine-123 or iodine-124).

As used herein, a “peptide” is from 2 to 25 (eg, as defined above) amino acid or peptide residues having one or more open valencies. This sequence may be linear or cyclic. For example, cyclic peptides can be prepared or produced by the formation of disulfide bridges. The peptides can be linked by the carboxy terminus, amino terminus, or other convenient point of attachment, such as, for example, the cysteine sulfur. Peptide derivatives are disclosed in US Pat. No. 4,612,30.
No. 2; 4,853,371 and 4,684,620 or as described in the Examples below. In particular, the peptide sequences referred to herein are written with the amino terminus on the left and the carboxyl terminus on the right.

The specific or specific values given below for radicals, substituents and ranges are for illustration only and do not exclude other defined values or other values within the specified ranges of radicals and substituents.

In particular, the peptides are 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.

As used herein, “residue of a compound of formula I” is a radical of a compound of formula I having one or more open valencies. If the resulting compound is capable of localizing near a tumor, one or more synthetically feasible atoms of the compound of Formula I will be removed to give an open valency. One skilled in the art can select, based on the desired linkage, appropriately functionalized starting materials obtained from compounds of Formula I using procedures known to those skilled in the art. For example, suitable molecules to be removed, a- carboxamide NH ₂ group of a hydrogen atom from the NH ₂ group of NH ₂ group or a- carboxamide (shown in FIG. 1), b-carboxamide (1) Or N of b-carboxamide
A hydrogen atom from the H ₂ group, the NH ₂ group of d-carboxamide (shown in FIG. 1) or d
- a hydrogen atom from the NH ₂ group of the carboxamide (1) e- carboxamide hydrogen atom from an NH ₂ group or e- carboxamide NH ₂ group of (1) and the 6 positions X, include Can be In addition, the hydroxyl group at the 3 'position of the sugar, the 3'
A hydrogen atom from the hydroxyl group at the position, a hydrogen atom of CH ₂ OH at the 5 ′ position, or a hydrogen atom from the hydroxyl group at the 5 ′ position of the sugar ring may be removed.

As used herein, “adenosyl” is an adenosine radical in which a synthetically available atom or group of atoms has been removed. As a synthetically feasible atom that can be removed, a hydrogen atom of a hydroxyl group at the 5 ′ position can be mentioned. Thus, adenosyl is
The 5 'position of adenosyl is conveniently linked to the 6 position of the compound of formula I.

As used herein, a “molecule comprising B-10” refers to one or more of B-10
Any compound containing an atom may be used. The nature of the molecule containing B-10 is not critical. However, the compounds are non-toxic and must be able to enter or localize near tumor cells when the atom containing B-10 is attached.

As used herein, “residue of a molecule comprising B-10” is a radical of a molecule comprising one or more open valencies. Synthesizable atoms or atoms of molecules, including B-10, can confer open valency as long as substantially biological activity is retained. One of skill in the art can use the procedures known to those skilled in the art to select a suitably functionalized starting material obtained from a compound containing B-10 based on the desired linkage.

As used herein, “residue of o-carborane”, “residue of m-carborane” or “residue of p-carborane” refers to o-carborane, m-carborane or p-residue, respectively. -A carborane radical, having one or more open valencies. One or more synthetically feasible atoms of o-carborane, m-carborane or p-carborane can provide open valency as long as substantially biological activity is retained. One of skill in the art can select an appropriately functionalized starting material obtained from o-, m- or p-carborane based on the desired linkage using procedures known to those skilled in the art.

Boron Compounds Useful for Neutron Capture Therapies The present invention provides compounds of formula I (FIG. 1) linked to one or more molecules, including B-10,
Where X is a molecule that contains CN, OH, CH _3, the adenosyl or B-10)
Or a pharmaceutically acceptable salt thereof.

Various molecules, including B-10, known to those of skill in the art are useful in the present invention. This molecule differs considerably in structure, but is suitable for practicing the present invention. Acceptable species include amino acids, carbohydrates, nucleosides, as well as boron, including carboranes. A wide range of boron-containing compounds are commercially available or known to those skilled in the art.
Various molecules including B-10 are available from Boron Bio, Raleigh, NC
logicals or RysCor Sci, Raleigh, NC
commercially available from ence.

In particular, one or more molecules comprising B-10 can be o-carborane, m-carborane or p-carborane. More specifically, one or more of the molecules, including B-10, is o-carborane. o-Carborane [1,2-dicarbadodecaborane (12)], m-carborane [1,7-dicarbadodecaborane (12)]; and p-carborane [1,12-dicarbadodecaborane (12)] Is commercially available from Aldrich, Milwaukee, Wisconsin.

In particular, each molecule comprising B-10 can independently be o-carborane, m-carborane or p-carborane. More specifically, each molecule containing B-10 has o
-Carborane.

Compounds of Formula I / Molecules Comprising B-10 The residue of the molecule comprising B-10 is an amide (eg, —N (R) C (＝ O) — or —C
(= O) N (R)-), ester (eg, -OC (= O)-or -C (= O) O-
), Ether (example -O-), amino (example -N (R)-), ketone (example -C (= O
)-), Thioether (example -S-), sulfinyl (example -S (O)-), sulfo
Nil (eg -S (O)₂-) Or by direct (e.g. CC bond) bonding
Can bind to the residue of the compound. Where each R is independently H or (C₁-C ₆ ) Alkyl. Such conjugation can be effected from appropriately functionalized starting materials.
It can be made by synthetic procedures known to those skilled in the art. The person skilled in the art
From the residues of the compounds of formula I using procedures known to those skilled in the art, and
Selects appropriately functionalized starting materials from residues of compounds containing B-10
can do.

The residue of the molecule comprising B-10 can be directly attached at a synthetically feasible position of the residue of the compound of formula I. Suitable attachment points include other attachment points, for example, b-carboxamide, d-carboxamide, e-carboxamide (FIG. 1).
), As well as the 6-position (the position occupied by X in FIG. 1), the 5′-hydroxyl group and the 3′-hydroxyl group of the 5-membered sugar ring. US Patent 5,739,31
3 is a compound (eg, cyanocobalamin-b- (4-aminobutyl) amide, methylcobalamin-b- (4-) which is a useful intermediate for preparing the compound of the present invention.
Aminobutyl) amide and adenosylcobalamin-b- (4-aminobutyl)
Amides).

Compounds in which the residue of the molecule comprising B-10 is attached at position 6 of the compound of formula I can reduce the corresponding Co (II) compound of formula I to form a nucleophilic Co (I) Prepared and converted to a Co (I) compound having a suitable leaving group such as a halide (eg, chloride).
Treatment with residues of the molecule containing -10 (or a derivative thereof).

The present invention also provides compounds having one or more residues of a molecule comprising B-10 directly attached to a compound of formula I. For example, the residue of the molecule comprising B-10 can be directly linked to the residue of b-carboxamide of the compound of formula I, and the residue of another molecule comprising B-10 is the d of the compound of formula I -It can bind directly to carboxamide residues.
In addition, the residue of the molecule containing B-10 can be directly attached to the 6 position of the compound of formula I, and the residue of another molecule containing B-10 is the d- or e of the compound of formula I -It can bind directly to carboxamide residues.

Compounds of Formula I / Linker / Molecules Containing the B-10 Linker The residues of the molecule containing B-10 not only bind directly to the residues of the compound of formula I, but also by a suitable linker It can also bind to a residue of the compound. The structure of the linker is not critical, provided that a compound of the invention is produced that has an effective therapeutic index for the target cell and is localized within or near the tumor molecule.

Suitable linkers include those that separate the residue of the compound of formula I from the residue of the compound comprising B-10 at about 5 to about 200 angstroms. Other suitable linkers include the residue of a compound of Formula I and the residue of a compound comprising B-10 from about 5 to about 1
The residue of the compound of formula I and B-10 together with a linker that separates at
Some linkers separate residues of a compound containing from about 5 to about 50 angstroms, or about 5 to about 25 angstroms. A suitable linker is described, for example, in US Pat.
, 735, 313.

The linker can be attached at any synthetically possible position of the residue of the compound of the formula I residue. Suitable points of attachment are, for example, residues of b-carboxamide, residues of d-carboxamide, residues of e-carboxamide, 6
In addition to the position (ie, the position occupied by X in the compound of formula I), the 5 ′ of the sugar five-membered ring
-Hydroxyl group and 3'-hydroxyl group. Those skilled in the art will be able to
Using procedures known to those skilled in the art, one can select suitably functionalized starting materials from the residues of the compounds of formula I and the residues of a given compound, including B-10.

The linker can be an amide (eg —N (R) C (＝ O) — or —C (＝ O) N (R) —
), Esters (e.g. -OC (= O)-or -C (= O) O-), ethers (e.g.-
O-), ketone (example -C (= O)-), thioether (example -S-), sulfinyl (example -S (O)-), sulfonyl (example -S (O) _2- ), amino (example) −N (R
)-) Or a direct (e.g. CC) bond to the residue of the compound of formula I or B-
10 can bind to residues of the molecule. Wherein each R is independently H or (C ₁ -C ₆ ) alkyl. The linkage can be made from appropriately functionalized starting materials by synthetic procedures known to those skilled in the art. One skilled in the art, based on the desired linkage, will use the procedures known to those skilled in the art to properly functionalize, from a given linker, the residue of the compound of Formula I, the residue of the compound comprising B-10. The selected starting material can be selected.

In particular, the linker may be a divalent radical of the formula WAQ, wherein A is (C ₁ -C ₆ ) alkyl, (C ₂ -C ₆ ) alkenyl, (C ₃ -C ₈ ) cycloalkyl or (C ₆ -C ₁₀ ) allyl, and W and Q are each independently —N (
R) C (= O)-, -C (= O) N (R)-, -OC (= O)-, -C (= O)
O-, -O-, -S-, -S (O)-, -S (O) _2- , -N (R)-, -C (
ＯO) — or a direct bond (ie, no W or Q), wherein each R is independently H or (C ₁ -C ₆ ) alkyl.

In particular, the linker can be a divalent radical of the formula W— (CH ₂ ) nQ, where n is about 1 to about 20, about 1 to about 15, about 2 to about 10, about 2 to about 6 or about 4 to
W and Q are each independently -N (R) C (= O)-, -C (=
O) N (R)-, -OC (= O)-, -C (= O) O-, -O-, -S-, -S
(O)-, -S (O) _2- , -C (= O)-, -N (R)-or a direct bond (i.e., no W or Q), wherein each R is independently H or (C ₁ -C ₆ ) alkyl.

Specifically, W and Q are each independently -N (R) C (= O)-, -C (=
O) N (R)-, -OC (= O)-, -N (R)-, -C (= O) O-, -O-
Or a direct bond (ie, no W or Q).

In particular, the linker is a divalent radical, ie a 1, ω-divalent radical generated from a peptide or an amino acid. The peptide can include 2 to about 20 amino acids, 2 to about 15 amino acids, or 2 to about 12 amino acids.

In particular, the peptide is a poly-L-lysine (ie, [—NHCH [(CH₂) ₄ NH₂] CO-] mQ, where Q is H, (C₁-C₁₄) Alkyl,
Or a suitable carboxy protecting group, wherein m is from about 2 to about 20. For more information,
Li-L-lysine can comprise from about 5 to about 15 residues (ie, m is about 5
~ 15). More specifically, poly-L-lysine has from about 8 to about 11 residues
(I.e., m is from about 8 to about 11).

In particular, the peptides may 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.

Specifically, the linker is 1,6-diaminohexane H₂N (CH₂)₆NH₂, 1
, 5-Diaminopentane H₂N (CH₂)₅NH₂, 1,4-diaminobutane H ₂ N (CH₂)₄NH₂Or 1,3-diaminopropane H₂N (CH₂)₃ NH₂It is prepared from

The linker can include one or more non-metallic radionuclides. In particular, the linker can include more than one non-metallic radionuclide. More specifically, the linker can include 2 to about 10, 2 to about 8, 2 to about 6, or 2 to about 4 non-metallic radioisotopes.

Certain residues of a peptide containing one or more non-metallic radionuclides (ie, linkers)
Is the following formula:

[0089]

Embedded image [Wherein independently for each M, non-metallic radionuclide, each R is _{independently,} (C _{1 -C} 1 _{4) alkyl,} (C 2 _{-C 14) alkenyl,} (C 2 _{-C 14)} alkynyl, (
C ₁ -C ₁₄₎ alkoxy, hydroxy, cyano, nitro, halo, _{trifluoromethyl, N (Ra) (Rb)} , (C 1 -C 14) _alkanoyl, (C 2 _{-C 14)} alkanoyloxy, _{(C 6} - C ₁₀ ) allyl or (C ₃ -C ₈ ) cycloalkyl, Ra and Rb are each independently H or (C ₁ -C ₁₄ ) alkyl, and P; Q is H, (C ₁ -C ₁₄₎ I) alkyl, or a suitable carboxy protecting group, n is 2 to about 20; I is 1 to 5, j is 0 to 4,
I + j is <= 5] or a pharmaceutically acceptable salt thereof.

Specifically, i is 1, j is 0, M is fluorine-18, bromine-76 or iodine-123, and n may be from about 6 to about 12.

A molecule comprising B-10 can contain one or more boron atoms. B-10
Suitable molecules comprising may contain 1 to about 20, 1 to about 15, 1 to about 10, or 1 to about 5 boron atoms. Further, atoms comprising B-10 can include one or more B-10 atoms. Suitable molecules including B-10 are from 1 to about 20
, 1 to about 15, 1 to about 10, or 1 to about 5 B-10 atoms.

[0092] The molecule comprising B-10 may be an amino acid, carbohydrate, nucleoside or carborane. In particular, the molecule comprising B-10 is o-carborane, m-carborane or p-carborane.

Compounds wherein the linker is attached to the 6-position of the compound of formula I prepare a nucleophilic Co (I) species as described herein above, which can be converted to an appropriate compound such as a halide (chloride). By reacting with a linker containing a suitable leaving group.

The present invention also provides compounds having one or more molecules comprising B-10, each linked to Formula I by a linker. For example, the residue of the molecule containing B-10 is conveniently linked to the residue of b-carboxamide of the compound of formula I by a linker.
Can be conveniently linked to the d- or e-carboxamide residue of the compound of formula I by a linker. In addition, the residue of the molecule containing B-10 is attached to the 6 position of the compound of formula I by a linker, and the residue of another molecule containing B-10 is attached to the b-, d of the compound of formula I by a linker. -Or e-carboxamide.

The present invention also provides compounds having one or more molecules comprising B-10 attached directly or via a linker to a compound of formula I. For example, the residue of a molecule comprising B-10 may be conveniently linked, directly or by a linker, to the residue of b-carboxamide of a compound of Formula I, and the residue of another molecule comprising B-10 may be directly or by a linker. , Formula I
To the d- or e-carboxamide residue of the compound of formula (I). In addition, the residue of the molecule comprising B-10 is attached, either directly or by a linker, to the 6 position of the compound of formula I, and the residue of another molecule comprising B-10 is attached, directly or by the linker, to the compound of formula I It can bind to a b-, d- or e-carboxamide residue.

Compounds of Formula I / Chelating Groups Containing Gadolinium 157 US Pat. No. 5,739,313 includes compounds of Formula I, a chelating group comprising a linking group, a detectable radionuclide or a paramagnetic ion. , And cobalamin analogs. Disclosed are compounds that localize within tumor cells after administration and are useful for tumor imaging.

[0097] Gadolinium 157, a metal radionuclide, is particularly useful for performing magnetic resonance imaging. It is also a useful target ion in neutron capture therapy. Applicants
One of the chelating molecules disclosed in US Pat. No. 5,739,313 is G
It has been found that the inclusion of d-157 can provide not only compounds that are particularly useful for performing magnetic resonance imaging, but also compounds that can be used in conjunction with neutron capture therapy to treat tumors.

Thus, the present invention provides a residue of a compound of formula I linked to one or more residues of the formula -QLW-Det, wherein each Det is independently Gd- 157, wherein each L is independently a linker (as defined above) or absent;
Each W and Q is independently -N (R) C (= O)-, -C (= O) N (R
)-, -OC (= O)-, -C (= O) O-, -O-, -S-, -S (O)-,
_{-S (O) 2 -, -} C (= O) -, - N (R) - or a direct bond (i.e. W also Q is empty), H or _(C 1 -C each R is independently ₆ ) It is alkyl.

Compounds of Formula I / Chelating Groups Containing Radionuclides Applicants have one or more neutron capture target atoms (eg, B-10 or Gd-
It has also been discovered that by incorporating 157) into compounds that also contain a detectable radionuclide, compounds useful for tumor imaging or therapy can be prepared. Thus, the present invention relates to a compound comprising B-10 comprising one or more residues linked to a compound of formula -QLWD
The residue of a compound of formula I, wherein each residue is also attached to one or more groups of et
Is independently a chelating group (as defined herein), including Gd-157;
Each L is independently a linker (as defined herein) or absent, and each W and Q is each independently -N (R) C (= O)-, -C (= O) N (R)-,-
OC (= O)-, -C (= O) O-, -O-, -S-, -S (O)-, -S (O
) ₂ —, —C (＝ O) —, —N (R) — or a direct bond (ie, W or Q is empty), and each R is independently H or (C ₁ -C ₆ ) alkyl Or a pharmaceutically acceptable salt thereof.

The present invention provides a residue of a compound of Formula I attached to a molecule comprising B-10 or to a chelating group comprising Gd-157. Wherein the residue of the compound of formula I is attached to one or more groups of the formula -QLW-Det, each Det independently being a chelating (as defined above) comprising a metal radionuclide. And each L is independently a linker (as defined above) or absent, and each W and Q is each independently -N
(R) C (= O)-, -C (= O) N (R)-, -OC (= O)-, -C (= O
) O-, -O-, -S-, -S (O)-, -S (O) _2- , -C (= O)-,-
N (R) - or a direct bond (i.e. W also Q is empty) is, each R is H or _(C 1 -C ₆₎ alkyl.

Compounds of Formula I / Molecules Containing B-10 / Detectable Radionuclides The present invention relates to compounds of formula I (FIG. 1) wherein 1) the residue of a molecule comprising B-10,
And 2) a compound bound to a detectable radionuclide, where X is CN, OH, CH ₃ , Adenosyl or a molecule comprising B-10], or a pharmaceutically acceptable
The salt is also provided.

The detectable radionuclide can be attached directly to the residue of the compound of formula I or by a linker to the residue of the compound of formula I. The linker may be any suitable linker described herein. Specifically, the linker is of the formula WA, where A is (
C ₁ -C ₆ ) alkyl, (C ₂ -C ₆ ) alkynyl, (C ₃ -C ₈ ) cycloalkyl, or (C ₆ -C ₁₀ ) allyl, and W is —N (R) C (＝ O )-,-
C (= O) N (R)-, -OC (= O)-, -C (= O) O-, -O-, -S-
_{, -S (O) -, -} S (O) 2 -, - C (= O) -, - N (R) - or a direct bond, each R is independently H or _(C 1 _-C 6) Alkyl and A is substituted by one or more non-metallic radionuclides. The linker may be a peptide or an amino acid.

Compounds of Formula I / Groups Containing Gd-157 / Detectable Radionuclides The present invention also provides that the residues of the compounds of Formula I (Figure I) include 1) a detectable radionuclide and 2) Gd-157. Or a pharmaceutically acceptable salt thereof is provided.

Gd-157 can be linked to a residue of a compound of Formula I in any suitable manner. For example, a residue of a compound of Formula I can be linked to a group comprising Gd-157 having the formula QLW-Det, wherein Det is a chelate group comprising Gd-157 and L is a linker Or absent, W and Q are each independently -N (R) C (
= O)-, -C (= O) N (R)-, -OC (= O)-, -C (= O) O-,-
O-, -S-, -S (O)-, -S (O) _2- , -C (= O)-, -N (R)-
Or a direct bond, wherein each R is independently H or (C ₁ -C ₆ ) alkyl. Any suitable chelator can be used.

The detectable radionuclide can be linked directly to the residue of the compound of formula I or can be linked to the residue of the compound of formula I by a linker. The linker can be any suitable linker described herein. Especially, the linker can be a formula W-A, the formula A _(C 1 -C _{6) alkyl, (C} 2 -C _{6) alkenyl, (C} 2 -C _{6) alkynyl, (C} 3 -C ₈ ) Cycloalkyl or (C ₆ -C ₁₀ ) aryl, wherein W is —N (R) C (＝ O) —, —C (＝ O) N (R) —, —OC (=
O)-, -C (= O) O-, -O-, -S-, -S (O)-, -S (O) _2- ,
—N (R) —, —C (＝ O) — or a direct bond, wherein each R is independently H or (C ₁ -C ₆ ) alkyl, wherein A is one or It has been replaced by more non-metallic radionuclides. The linker can also be a peptide or an amino acid.

Non-Metallic Radionuclides Any detectable radionuclide that is suitable for imaging can be used in the compounds of the present invention. For example, suitable non-metallic radionuclides include carbon-11, fluorine-18, bromine-76 and iodine-123. In particular, the non-metallic radionuclide is a non-metallic paramagnetic atom (eg, fluorine-19) or a non-metallic positron-emitting radionuclide (eg, carbon-11, fluorine-18, iodine-123 or bromine-7).
6).

Metal Radionuclides Suitable metal radionuclides (ie metal radioisotopes or metal paramagnetic ions)
Include antimony-124, antimony-125, arsenic-74, barium-10
3, 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, hafnium-175, hafnium-
175-181, holmium-166, indium-110, indium-11
1, iridium-192, iron-55, iron-59, krypton-85, lead-210
Manganese-54, mercury-197, mercury-203, molybdenum-99, neodymium-147, neptunium-237, nickel-63, niobium-95, osmium-185 + 191, palladium-103, platinum-195m, praseodymium-14
3, promethium-147, protactinium-233, radium-226, rhenium-186, rhenium-188, rubidium-86, ruthenium-103,
Ruthenium-106, scandium-44, scandium-46, selenium-75
, Silver-110m, silver-111, sodium-22, strontium-85, strontium-89, strontium-90, sulfur-35, tantalum-182, technitium-99m, tellurium-125, tellurium-132, thallium-204, thorium. -228, thorium-232, thallium-170, tin-113, tin-
114, tin-117m, titanium-44, tungsten-185, vanadium-48, vanadium-49, ytterbium-169, yttrium-86,
Yttrium-88, Yttrium-90, Yttrium-91, Zinc-65 and Zirconium-95 are included.

As used herein, a “detectable radionuclide” is any suitable radionuclide (ie, radioisotope) that can detect cancer or other neoplastic cells in an in vivo or in vitro diagnostic procedure. ). Suitable detectable radionuclides include metal radionuclides (ie, metal radioisotopes) and nonmetal radionuclides (
That is, non-metal isotopes).

As used herein, “therapeutic radionuclide” is any suitable radionuclide (ie, radioisotope) that has a therapeutic effect on cancer or other neoplastic cells in vivo or in vitro. Body). Suitable therapeutic radionuclides include metal radionuclides (ie, metal radioisotopes).

Chelating Groups Any suitable chelating group can be incorporated into the compounds of the invention. Suitable chelating groups include those disclosed in US Pat. No. 5,739,313. In particular, the chelating groups are NTA, HEDTA, DCTA, RP414, M
DP, DOTATOC, CDTA, HYNIC, EDTA, DTPA, TETA
, DOTA, DOTMP, DCTA, 15N4, 9N3, 12N3 or MAG
3 (or other suitable polyamino acid chelators), which are described herein below, or may be phosphate chelators (eg, EDMT). More particularly, the chelate group can be DTPA.

DTPA is dimethylenetriaminepentaacetic acid and 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, and 15N4 is 1,4,8,12-tetraazacyclopentadecane-N, N ′, N ″, N ′ ″-tetraacetic acid, 9
N3 is 1,4,7-triazacyclononane-N, N ′, N ″ -triacetic acid, and 12N3 is 1,5,9-triazacyclododecane-N, N ′, N ″ -triacetic acid. Acetic acid, MAG3 is (N- [N- [N-[(benzoylthio) acetyl] glycyl] glycyl] glycine), and DCTA is of the formula

[0112]

Embedded image Wherein R ³ is (C ₁ -C ₄ ) alkyl or CH ₂ CO ₂ —, wherein R ³ is attached via the 4- or 5-position or via the R ³ group Wherein R ³ is one to four detectable metal or nonmetal cations (
M), a monovalent cation, or an alkaline earth metal. Thus, with a metal in oxidation state +1, each individual cyclohexane-based molecule may have up to four metal cations, where both R ³ groups are CH ₂ COOM.
Furthermore, at higher oxidation states, the number of metals will decrease to 2 or 1 per cyclohexane skeleton. This formula is not intended to limit the molecule to any particular stereochemistry.

[0113] NTA, HEDTA and DCTA are available from Poster Sessions, Pr.
receiveds of the 46th Annual Meeting
, J. et al. Nuc. Med. , P. 316, no. 1386. RP
414, Scientific Papers, Proceedings o
f the 46th Annual Meeting, J. Am. Nuc. Med.
, P. 123, No. 499. MDP is Scientific
c Papers, Proceedings of the 46th Ann
ual Metting, J.A. Nuc. Med. , P. 102, no. 413. DOTATOC is available from Scientific Papers, P
rosedsing of the 46th Annual Metin
g. Nuc. Med. , P. 102, no. 414 and Scientif
ic Papers, Proceedings of the 46th An
Null Metting, J. et al. Nuc. Med. , P. 103, no. 415
Is disclosed. CDTA is from Poster Sessions, Proce
edings of the 46th Annual Meeting, J. Am.
Nuc. Med. , P. 318, no. 1396. HYNIC
Is the Poster Sessions, Proceedings of the
46th Annual Meeting, J. Am. Nuc. Med. , P. 31
9, No. 1398.

Bifunctional chelators (ie, chelating groups) based on macrocyclic ligands in which conjugation is via an activation arm attached to the ligand's carbon skeleton are also described in M.A. Moi et al. Amer. Chem. , Soc. , 49, 2639 (1
989) (2-p-nitrobenzyl-1,4,7,10-tetraazacyclododecane-N, N ′, N ″, N ′ ″-tetraacetic acid), S.P. V. Deshpanda
e et al. Nucl. Med. , 31, 473 (1990); Kuser et al., Bioconj. Chem. , 1,345 (1990); J. Broan
Et al. C. S. Chem. Comm. , 23, 1739 (1990), and C.I. J. Anderson et al. Nucl. Med. 36,850 (1995)
) (6-bromoacetamide-benzyl-1,4,8,11-tetraazacyclotetoadecane-N, N ′, N ″, N ′ ″-tetraacetic acid (BAT)) Can be used as a base.

In addition, diagnostic or therapeutic chelators can be used with Scientific
Papers, Proceedings of the 46th Annu
al Metting, J. et al. Nuc. Med. , Wednesday, June
9, 1999, p. 124, no. It can be any of the chelating groups disclosed at 500.

A “detectable chelating group” is a chelating group that includes a metal radionuclide (eg, a metal radioisotope) that can detect cancer or other neoplastic cells in vivo or in vitro.

Among other things, chelating groups are described in Waibel et al., Nature Biotechn.
ology, 897-901, Vol. 17, September 1999 or Sattelberger et al., Nature Biotechnology.
, 849-850, Vol. 17, any one of the carbonyl conjugates disclosed in September 1999.

Among other things, detectable chelates are described by Waibel et al., Nature Bio.
technology, 897-901, Vol. 17, September
1999 or Sattelberger et al., Nature Biotechn.
ology, 849-850, Vol. 17, any of the carbonyl complexes disclosed in September 1999, which may further include a metal radionuclide. More particularly, the groups of detectable chelates include Wabel et al., Na
cure Biotechnology, 897-901, Vol. 17, Se
ptember 1999 or Sattelberger et al., Nature
Biotechnology, 849-850, Vol. 17, Septemb
er 1999, which may further include technitium-99m.

As used herein, “therapeutic chelating group” refers to a metal radionuclide (eg, a metal radioisotope) that has a therapeutic effect on cancer or other neoplastic cells in vivo or in vitro. Chelating groups. Any suitable chelating group can be used.

In particular, therapeutic chelating groups are described in Waibel et al., Nature Biote.
channel, 897-901, Vol. 17, September 19
99 or Sattelberger et al., Nature Biotechnol
ogy, 849-850, Vol. 17, any of the carbonyl complexes disclosed in September 1999, which may further include a metal radionuclide. More particularly, therapeutic chelating groups are described in Waibel et al., Nature.
Biotechnology, 897-901, Vol. 17, Septem
ber 1999 or Sattelberger et al., Nature Biot
technology, 849-850, Vol. 17, September 1
999, further comprising rhenium-186.
Or it may contain rhenium-188.

A tumor that can be treated with the compounds and methods of the present invention can be located in any part of a mammal. Among others, tumors can be found in the breast, lung, thyroid, lymph nodes, genitourinary (eg, kidney, ureter, bladder, uterus, ovary, or prostate), musculoskeletal system (eg, bone, skeletal muscle or bone marrow), gastrointestinal tract (eg, Stomach, esophagus, small intestine, large intestine, duodenum, pancreas,
Liver or smooth muscle), central or peripheral nervous system (eg brain, spinal cord or nerve),
It may be localized to a head and neck tumor (eg, ear, eye, nasopharynx, oropharynx, or salivary gland) or heart.

The compounds disclosed herein are prepared using procedures similar to those described in US Pat. No. 5,739,313 or using procedures similar to those described herein. it can. The residue of the molecule comprising B-10 can be linked to the residue of a compound of formula I as described herein above. Additional intermediates and synthetic procedures that are useful for the preparation of compounds of the present invention are described, for example, in Hogenkamp, H. et al., Synthesis and Characterization of Nido-Carborane-Cobalamin Conjugates (Synthesis and
Characterization of nido-Carborane-
Cobalamine Conjugates), Nucl. Med. & Bio
l. , 2000, 27, 89-92; Collins, D .; , Et al. Tumor Imaging Via Indium 111-Labeled DTPA-Adenosylcobalamin (Tumor Imaging Via Indium 111-Labe)
led DTPA-Adenosylcobalamine), Mayo Cli
nic Proc. No. 60 / 129,733 filed on Apr. 16, 1999; U.S. Pat. No. 60 / 129,733 filed on Oct. 15, 1999. No. 159,874, U.S. Pat. No. 60 / 159,753, filed Oct. 15, 1999;
No. 60 / 159,873, filed Oct. 15, 1999, and the references cited therein.

Where the compound is sufficiently basic or acidic to form stable nontoxic acid or base salts, administration of the compound as a salt may be preferred. Examples of pharmaceutically acceptable salts are pharmaceutically acceptable anions such as tosylate, methanesulfonate, acetate, citrate, malonate, tartrate, succinate, benzoate, ascorbate, α-ketoglutarate and α-ketoglutarate. It is an organic acid addition salt formed with an acid forming glycerophosphate. Suitable non-organic salts may also be formed including hydrogen chloride, sulfuric acid, nitric acid, bicarbonate and carbonate.

Pharmaceutically acceptable salts are prepared by reacting a sufficiently basic compound, such as an amine, with a suitable acid that provides a physiologically acceptable anion, by standard procedures known to those skilled in the art. May be obtained by using Alkali metal (eg, sodium, potassium or lithium) or alkaline earth metal (eg, calcium) salts of carboxylic acids can also be made.

The compounds of the present invention can be formulated as pharmaceutical compositions and administered to human patients in a variety of forms adapted to the chosen route of administration, ie, oral or parenteral (eg, intravenous, intramuscular, intraperitoneal). Such mammalian hosts can be administered. Preferably, the compounds are administered parenterally.

The active compounds 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 non-toxic 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.

Pharmaceutical dosage forms suitable for injection or infusion include sterile aqueous solutions or dispersions or sterile injectable or infusible solutions or active ingredients, optionally in liposomes, which are compatible with the extemporaneous preparation of dispersions. May be included. In all cases, the final dosage form should be sterile, fluid and stable under the conditions of manufacture and storage. The liquid carrier or excipient can be a solvent or liquid dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, liquid polyethylene glycol, and the like), vegetable oils, non-toxic glyceryl esters, and suitable mixtures thereof. Proper fluidity can be maintained, for example, by formation of liposomes, by retention of the required particle size in the case of dispersion, or by use of surfactants. Prevention of the action of microorganisms can be achieved by treating various antimicrobial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid,
This can be done by something like thimerosal. In many cases, it may be preferable to include isotonic agents, for example, sugars, buffers or sodium chloride. Absorption of the injectable compositions can be delayed by using agents in the composition that delay absorption such as, 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 the various other ingredients enumerated above and, if necessary, then sterilizing by filtration.
In the case of powders for the preparation of a sterile injectable solution, the preferred preparation methods are suction drying and lyophilization techniques, which yield a powder of the active ingredient plus any further desirable ingredients already present in the sterile filtered solution.

For illustration, suitable administration of a compound of the invention for use in therapy in combination with a neutron capture will range from about 0.1 μg to about 100 μg per treatment, for example about 0.5 μg Administration in the range of from about 50 μg to about 50 μg, or about 0.5 μg to 15 μg is included. Suitable dosages for use in imaging, or for use in imaging and therapy, range from about 0.1 mg to about 50 g, e.g. Dosages in the range of 5 mg to about 10 g, or about 0.5 g to 2 g, are included.

The desired administration may be in the conventional manner in a single dose or in preferred intervals, for example, as two, three, four or more sub-doses per day. The sub-dose itself may be further divided, eg, into a number of discrete suitably spaced doses.

The compound is preferably dissolved or dispersed in the non-toxic solution carrier, eg, a saline or similar aqueous carrier, at the desired concentration. The therapeutic unit dose selected above is then administered to the test animal or human patient by oral administration or ingestion, or by parenteral administration such as intravenous or intraperitoneal infusion or injection to achieve the desired in vivo concentration. Administrations useful for treating tumors include:
In vitro, as described herein below those known to be effective in treating tumors in a human or an animal model, or animal treatment other already used in the labeled vitamin B ₁₂ molecules It can be derived from administration.

FIGS. 2 and 3 each illustrate a four-step synthesis used to prepare three cyanocobalamin-nido-carborane conjugates. o-Carboranecarboxylic acid (2) was prepared by reacting o-carborane with n-butyllithium in ether and carboxy dioxide at -78 ° C for approximately 1 hour. Treatment with thionyl chloride affords o-carborane carboxylic acid chloride (3), which is reacted with 1,4-butanediamine in pyridine to bind amide (4) to nido-carboranoyl (4-aminobutyl) amide. I got Compound (4) was coupled with the b-monocarboxylic acid of cyanocobalamin, the d-monocarboxylic acid of cyanocobalamin, or both the b- and d-dicarboxylic acids of cyanocobalamin. In each reaction, hydroxybenzotriazole and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide were added to promote amide bond formation.

The present invention will be further illustrated by the following examples.

Examples Ultraviolet-visible spectra were recorded on a diode array spectrophotometer. The ¹¹ B NMR spectrum at 192.656 MHZ by ¹ H decoupling was 8 m
Varian 1NOVA 600 MHz with m broadband probe
Recorded on a spectrometer. Approximately 10 mg of cobalamin-conjugate was analyzed by 5 mm NMR.
It was dissolved in pyridine _{d 6} of 1mL in a tube. 500 scans to 0.13
Collected with a 3 second ring capture time and a 1 second relaxation delay. Chemical shift to BF ₃ :(
CH ₃ CH ₂ ) ₂ O.

[0135] Mass spectrometry data was obtained on a Sciex API 365 LC-MS / MS system (Toronto, Canada). The separation was performed using two LC-10AD pumps and one SCL-10Avp controller (Shimadzu Scientific Instruments, Shima).
zu Scientific Instruments, Columbia, M
D) on a Shimazu HPLC system. Analyze A
Monitored with UV at 214 NM on a BI785 detector. HPLC separation, BD
S-Hypersil C8 column (150 × 4.6 mm, 120A, Kellstone Scientific, Bellefonte)
, PA). The mobile phase was H ₂ O: methanol (98:
2 v: v) and pump B H ₂ O: methanol (2:92 v: v). A linear gradient from 5% B to 30% B over 10 minutes was used and held at 30% B for 20 minutes before returning to initial conditions (mono-carborane synthesis). Dicarborane requires a longer gradient with a similar mobile phase, which is 5 minutes over 25 minutes.
% B to 65% B and hold at 65% B for 15 minutes before returning to initial conditions.
Separation was monitored by UV absorption at 214 NM. The flow rate was 1.0 ml / min, a post-column slit that allowed 〜１０10 μl to flow into the mass spectrometer.

Mass spectral data were collected using electrospray ionization in the positive mode with a dwell time of 0.3 ms / 0.1 AMU in a mass range of 300-2300 AMU. Synthetic samples were prepared at 5 or 10 mg / ml in Pump A mobile phase and some were HP
Injected into LC (1-5 μl). The 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 performed by collecting fractions at elution times for several injections. The collected fractions were combined and dried to a powder. A part of the purified product was dissolved in methanol: H ₂ O (1: 1) and re-analyzed by HPLC-MS to confirm the purity.

O-Carborane, butyllithium (1.6 M solution in hexane) and putrescine were purchased from Aldrich Chemical Company.
ny, Milwaukee, WI). Water-soluble carbodiimide, 1-
Ethyl-3 (3'-dimethylaminopropyl) carbodiimide and 1-hydroxybenzotriazole were purchased from Sigma (St. Louis, MO). Thin layer chromatography (TLC) silica gel plates were obtained from Eastman Kodak Company. Cyanocobalamin-b, d and e monocarboxylic acids and b, d-dicarboxylic acids were prepared as previously described (U.S. Patent No. 5,739,313 and D.L.
Anton, et al. , J. et al. Am. Chem. Soc. , 1980, 102
, 2212-2219), courtesy of Dr. Kahl of the School of Medical Chemistry, University of California, San Francisco.

Example 1 Cyanocobalamin-nido-carborane conjugate (5, FIG. 2) 1 g (〜0.66 mmol) in 100 mL of a water-acetone mixture (2: 1)
B-monocarboxylic acid, d-monocarboxylic acid or b, d- of cyanocobalamin
Dicarboxylic acid, hydroxybenzotriazole (810 mg, 6.0 mmol)
, 1-ethyl-3 (3-dimethylaminopropyl) carbodiimide (11.4 g)
, 6.0 mmol) and 4 (600 mg, 2.0 mmol) were adjusted to pH 6.9 with 1N NaOH. The reaction was stirred at room temperature and its progress was monitored by TLC. After 3 hours, the mixture was concentrated to remove acetone, and the resulting aqueous suspension was extracted into 92% aqueous phenol. The phenol phase was washed with water to remove water-soluble reactants. One volume of acetone and three volumes of ether were added to each phenol phase, and the desired cyanocobalamin-nide-carborane conjugate was re-extracted into water. The aqueous layer was extracted three times with ether to remove residual phenol and unreacted (4). Finally, the aqueous solution was evaporated and dried.
The residue was triturated with acetone and the desired conjugate was isolated as a light-red powder (90-95% yield based on cyanocobalamin-carboxylic acid).

The isotope ratio of the final product was also compared to theory as follows. Theoretical value of product M
+ H ⁺ (C70H109CoB9O15PN15) -1584.8 (12%),
1585.8 (37%), 1586.8 (74%), 1587.8 (100%)
, 1588.8 (84%), 1589.8 (44%), 1590.8 (16%)
, 1591.8 (4%). Synthetic I-1584.8 (14%), 1585.8 (3
75.8), 1586.8 (74%), 1587.8 (100%), 1588.8 (
88%), 1589.8 (48%), 1590.8 (19%), 1591.8 (
6%). Synthesis II-1584.8 (15%), 1585.8 (40%), 158
6.8 (78%), 1587.8 (100%), 1588.8 (86%), 15
89.8 (44%), 1590.8 (17%), 1591.8 (7%).

Diproduct theoretical value M + H ⁺ − (C77H129CoB18O16PN16) -18
15.1 (9%), 1816.1 (23%), 1817.1 (47%), 181
8.1 (77%), 1819.1 (99%), 1820.1 (100%), 18
21.1 (77%), 1822.1 (44%), 1823.1 (19%), 18
24.1 (6%). Measured value synthesis-1815.1 (15%), 1816.1 (27
%), 1817.1 (51%), 1818.1 (79%), 1819.1 (10%
0%), 1820.1 (100%), 1821.1 (78%), 1822.1 (
49%), 1823.1 (25%), 1824.1 (12%).

Results show b and d carborane cyanocobalamin (CCC) by LC-MS
Analogue identification is also included. The identified products were subsequently separated by HPLC and purified. The purified product was re-analyzed by LC-MS for detection of starting material. MS
/ MS data was also obtained for the b and d CCC analogs and further structural characterization was performed. These purified products were then tested in a biological assay. e-CCC
Analogs and di-CCC analogs were also separated and identified by LC-MS, however, purification was difficult because the preparation contained more reaction by-products. The isotope ratios of the b- and d-CCC and di-CCC analogs were compared with the theoretical isotope ratios to provide further information on identification.

The mass spectrum was checked for the presence of cyanocobalamin-nido-carborane conjugate. Mass spectrometry analysis and ¹¹ B NMR showed that during the conversion of 3 to 4 (FIGS. 2 and 3), the o-carborane nucleus lacked the boron atom, yielding the nido-carborane derivative 4 (FIG. 3). showed that. UV-visible spectroscopy of the final product shows the largest typical of cyanocobalamin, which retains the choline nucleus,
This suggests that the 5,6-dimethylbenzimidazole nucleotide is still attached to the cobalt atom.

Starting material 4 was prepared as follows (see FIG. 2).

A. o-Carboranecarboxylic acid (2). 500m in a 1L round bottom flask
A solution of o-carborane (1) (5.0 g, 34.7 mmol) in 1 dry ether was cooled in a dry ice-acetone bath to -78 ° C. The solution was slushed with argon, and the flask was sealed with a seal stopper. n-butyl lithium (2
4 mL, 1.6 M in hexane) was slowly injected via syringe over about 20 minutes and the reaction was stirred for another 30 minutes. Then crushed dry ice (10-15
g) was added and the reaction was stirred for 1 hour. The dry ice-acetone bath was removed and the reaction was allowed to come to room temperature. Ether and hexane were removed on a rotary evaporator, water (150 ml) was added, and unreacted o-carborane was removed from hexane (2 × 1).
00 ml). The aqueous phase was acidified with concentrated HCl and the desired product was extracted with hexane (4 × 100 ml). The extracts were combined, dried over _Na 2 SO _4, evaporated to dryness, to give a 5.8 g (88.7%) of o- carborane carboxylic acid (2), used without further purification did.

B. O-Carborane carboxylic acid chloride (3). P ₂ O ₅ on at dried o- carborane carboxylic acid (2.0 g, 10.6 mmol) was dissolved in thionyl chloride 30 ml, it was heated under reflux for 3 hours. The solution was cooled to room temperature, evaporated to _dryness, dried over P 2 _{O 5,} was used to obtain a (3) (2.05g, 9.9mmol) , without further purification.

C. Nido-carboranoyl (4-aminobutyl) amide (4). The acid chloride (3) is dissolved in 15 ml of dry pyridine and 1,4-diaminobutane (1.1
2 g, 12.7 mmol) were added. The reaction mixture was heated under reflux for 3 hours, cooled to room temperature and concentrated. Water (10 ml) was added and the suspension was acidified with 3M HCl. The desired product was extracted with ethyl acetate (4 × 50 ml), the combined organic layers were washed once with water, dried over Na ₂ SO ₄ , evaporated to dryness and 2.3.
5 g (8.0 mmol, 75%) of (4) were obtained. Therefore, the product is resistant to crystallization from various solvent mixtures, but TL on silica gel plates
C (2-propanol _{-NH 4 OH-H 2 O;} 7: 1: 2) by only one ninhydrin positive compounds differ from the diamine was shown.

Example 2 6-carboranoylamidopropyl cobalamin 6- (3-aminoprop-1-yl) cobalamin (300 mg), hydroxybenzotriazole (270 mg), 1-ethyl-3- (3-dimethylaminopropyl) Carbodiimide (382 mg) and o-carborane monocarboxylic acid were dissolved in water (50 ml) and acetone (20 ml) and reacted at room temperature for 3 hours. The reaction mixture was concentrated to remove acetone and desalted via phenol extraction. The concentrated aqueous solution was crystallized from aqueous acetone to obtain the title compound.

The intermediate 6- (3-aminoprop-1-yl) cobalamin was prepared as follows.

A. 6- (3-Aminoprop-1-yl) cobalamin. Cyanocobalamin (5
00 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. After 30 minutes, 3-chloropropylamine hydrogen chloride (130 mg) dissolved in 5 ml of deoxylated ethanol was added. One hour later, at room temperature, the mixture was desalted by phenol extraction. Aminopropylcarboramine was added to one volume of acetone and 3 volumes.
After addition of a volume of ether, it was re-extracted into water. Concentrate the aqueous solution, desired 6
-(3-Aminoprop-1-yl) cobalamin was crystallized from aqueous acetone (
Yield 510 mg).

Example 3 DTPA-Aminopropylcarbalamine (DAPC) Using a coupling procedure similar to that described in Example 2, 6- (3-aminoprop-
1-yl) carbalamine and DTPA were coupled in the presence of hydroxybenzotriazole to give the title compound.

Example 4 In Vitro Biological Activity of Carborane Cyanocobalamin Analogs Assess the in vitro binding of carborane cyanocobalamin (Example 1, CCC) and DTPA-aminopropylcobalamin (Example 3, DPAC) analogs to the transcobalamin protein. to the non-saturated vitamin _{B 12} binding capacity (UB
BC) Assay (DA, Collins and HPC Hogen)
kamp, J .; Nuclear Medicine, 1997, 38, 717-
723) was performed. Serum from Mayo Clinic
) Obtained from 5 patients evaluated for pernicious anemia. The patient's serum was first subjected to the conventional clinical UBBC described above. The CCC and DAPC analogs are Co-
Excess sera from five patients were subjected to modified UBBC when it was possible to inhibit the binding of 57 cyanocobalamin to the transcobalamin protein.

Specifically, under dim light, 0.4 ml of serum was added to 4 μl (concentration 10 μg C
(CC / ml normal saline) was treated with two CCC analogs, carborane-d-cyanocobalamin and carborane-b-cyanocobalamin, and DAPC. Analogs were incubated with patient serum for 20 minutes at room temperature. Clinical and analog-treated samples were assayed for UBBC as usual.

Analogs competitively inhibited Co-57-cyanocobalamin by binding to the transcobalamin protein. Therefore, the cp of the modified UBBC assay
m was significantly lower than that of the clinical sample. The percent PB of the analog relative to the transcobalamin protein was calculated as follows (PB = 100-CC
C _UBBC cpm / clinical UBBC cpm × 100). The mean percent binding (PB) of the five solutions to carborane-d cyanocobalamin and carborane-b cyanocobalamin (n = 10 for each solution, ie two modified UBBC assays per patient) was 35.75% and 92.93, respectively. %
And 98.21% for DAPC.

All publications, patents and patent texts are individually incorporated by reference.
It is incorporated herein by reference. The present invention has been described in terms of various specific and preferred embodiments and techniques. However, it should be understood that many changes and modifications can be made while remaining within the spirit and scope of the invention.

[Brief description of the drawings]

FIG. 1 shows the structure of cobalamin, where X is CN (cyano), OH, CH or adenosyl.

FIG. 2 shows the synthesis of cyanocobalamin-nido-carborane conjugate.

FIG. 3 shows the synthesis of representative compounds of the invention.

──────────────────────────────────────────────────続 き Continuation of front page (81) Designated country EP (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE ), OA (BF, BJ, CF, CG, CI, CM, GA, GN, GW, ML, MR, NE, SN, TD, TG), AP (GH, GM, KE, LS, MW, SD, SL, SZ, TZ, UG, ZW), EA (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), AE, AG, AL, AM, AT, AU, AZ, BA, BB, BG, BR, BY, CA, CH, CN, CR, CU, CZ, DE, DK, DM, DZ, EE, ES, FI, GB, GD, GE, GH, GM, HR , HU, ID, IL, IN, IS, JP, KE, KG, KP, KR, KZ, LC, LK, LR, LS, LT, LU, LV, MA, MD, MG, MK, MN, MW, MX, NO, NZ, PL, PT, RO, RU, SD, SE, SG, SI, SK, SL, TJ, TM, TR, TT, TZ, UA, UG, US, UZ, VN, YU, ZA , ZW (72) Inventor Collins, Douglas Ay United States, Minnesota 55902, Rotiester, Meadowlark Court Southwest 1150 (72) Inventor Hogenkamp, Henrikus PC, United States, Minnesota 55113, Lo Zubil, Marion Road, 2211 F-term (reference) 4C084 AA12 NA14 ZB262 4C085 HH03 JJ11 KA29 KB39 KB56 LL18 4C086 AA01 AA02 CB14 MA01 MA04 NA14 ZB26

Claims (74)

[Claims] 1. The residue of the compound of formula I (FIG. 1) is linked to the residue of a molecule comprising B-10, wherein X comprises CN, OH, CH ₃ , adenosyl or B-10. A compound, or a pharmaceutically acceptable salt thereof, that is a molecule. 2. The compound of formula I wherein the residue of the molecule comprising B-10 is directly attached to position 6 of the compound of formula I or which is directly attached to the residue of the b, d or e-carboxamide group of the compound of formula I The compound of claim 1, wherein: 3. The residue of the molecule comprising B-10 is directly attached to position 6 of the compound of formula I or directly to the residue of the a, b, d or e-carboxamide group of the compound of formula I The compound of claim 1, wherein the compound is: 4. The compound according to claim 1, wherein the residue of the molecule comprising B-10 is attached to the residue of the b-carboxamide group of the compound of formula I. 5. The compound according to claim 1, wherein the residue of the molecule comprising B-10 is attached to the residue of the d-carboxamide group of the compound of formula I. 6. The compound of claim 1, wherein the residue of the molecule comprising B-10 is attached to the residue of the e-carboxamide group of the compound of formula I. 7. The residue of the molecule containing B-10 is linked to the residue of the b-carboxamide group of the compound of formula I, and the second residue of the molecule containing B-10 is linked to the d-carboxamide group. 2. The compound of claim 1, wherein the compound is attached to a residue of 8. The compound according to claim 1, wherein the residue of the molecule comprising B-10 is attached at position 6 of the compound of formula I. 9. The compound of claim 1, wherein the molecule comprising B-10 contains 1 to up to about 20 boron atoms. 10. The compound according to claim 1, wherein the molecule comprising B-10 is an amino acid, carbohydrate, nucleoside or carborane. 11. The compound according to claim 1, wherein the molecule comprising B-10 is o-carborane, m-carborane or p-carborane. 12. The compound of claim 1, wherein the molecule comprising B-10 is o-carborane. 13. The method of claim 12, wherein the one or more linkers are of the formula WAQ, wherein A is (C ₁ -C₆) Alkyl, (C₂-C₆) Alkenyl, (C₂-C₆) Alkynyl,
(C₃-C₈) Cycloalkyl, or (C₆-C₁₀) Allyl, where
W and Q are independently -N (R) C (= O)-, -C (= O) N (R)-,
-OC (= O)-, -C (= O) O-, -O-, -S-, -S (O)-, -S (
O)₂-, -N (R)-, -C (= O)-or a direct bond, wherein each R is
Independently H or (C₁-C₆4.) The compound of claim 3, which is an alkyl. 14. W is NH ₂ or COOH and Q is NH ₂ or COOH.
14. The compound according to claim 13, which is OH. 15. The compound according to claim 13, wherein A is (C ₁ -C ₆ ) alkyl. 16. The compound of claim 3, wherein the one or more linkers is no less than about 5 angstroms and no more than about 50 angstroms. 17. The compound of claim 3, wherein the one or more linkers comprises a therapeutic or diagnostic radionuclide. 18. The compound according to claim 17, wherein the therapeutic radionuclide is a metal radionuclide. 19. The compound according to claim 17, wherein the diagnostic radionuclide is a metal radionuclide. 20. The therapeutic radionuclide is a non-metallic radionuclide.
The compound according to the above. 21. The compound of claim 3, wherein the one or more linkers is a divalent radical generated from a peptide. 22. The compound of claim 3, wherein the one or more linkers is a divalent radical generated from an amino acid. 23. The method according to claim 20, wherein the at least one linker is 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 4. The compound of claim 3, which is -tyrosine. 24. The compound of claim I, wherein the residue of the compound of formula I is also linked to a linker containing a detectable or therapeutic radionuclide. 25. A molecule wherein the residue of the compound of formula I (FIG. 1) is attached to a group of formula QLW-Det, wherein X is CN, OH, CH ₃ , adenosyl, B-10. ,
Or QLW-Det, wherein Det is a chelating group containing Gd-157, L is or is not a linker, and W and Q are each independently -N (R
) C (= O)-, -C (= O) N (R)-, -OC (= O)-, -C (= O) O
-, -O-, -S-, -S (O)-, -S (O) _2- , -C (= O)-, -N (
R) - or a direct bond, each R is H or (C 1 _{-C 6)} alkyl independently, a compound or a pharmaceutically acceptable salt thereof,. 26. A group of the formula QLW-Det is present at the residue of the b-carboxamide group, the residue of the d-carboxamide group, the residue of the c-carboxamide group or the 6-position of the compound of the formula I 26. The compound of claim 25, wherein the compound is attached. 27. A group of the formula QLW-Det is attached to the residue of the b-carboxamide group of the compound of the formula I, and the second group of the formula QLW-Det is a compound of the formula I Of d
26. The compound according to claim 25, which is attached to the residue of a carboxamide group. 28. The compound of claim 25, wherein the group of formula QLW-Det is about 20 to about 500 angstroms or less in length. 29. The method wherein the one or more chelating groups is EDTA, DTPA, DOTA
26. The compound of claim 25, which is DOTMP, TETA, MAG3 or DCTA. 30. The compound of claim 25, wherein said one or more chelating groups is DPTA comprising Gd-157. 31. The residue of the compound of formula I (FIG. 1) is attached to the residue of a molecule comprising B-10, and the residue of the compound of formula I is also a group of formula QLW-Det. X is C
N, OH, CH ₃ , adenosyl, a group of formula QLW-Det, or a molecule comprising B-10, wherein Det is a chelating group comprising a therapeutic or diagnostic radionuclide; Is or is not a linker, and Q and W are each independently -N (R) C (= O)-, -C (= O) N (R)-, -OC (= O)-, -C
(= O) O-, -O-, -S-, -S (O)-, -S (O) _2- , -C (= O)
—, —N (R) — or a direct bond, wherein each R is independently H or (C ₁ -C ₆ )
A compound which is alkyl, or a pharmaceutically acceptable salt thereof. 32. One or more radionuclides is ^{Tc 99m, In 111, In 1} 10, Gd 157 or ^{Y 86,} compound of claim 31. 33. The compound of claim 31, wherein the molecule comprising B-10 is attached to the compound of formula I at the residue or 6-position of the b-carboxamide, d-carboxamide or c-carboxamide group. . 34. The chelating group is DTA, DTPA, DOTA, TETA,
The compound according to claim 31, which is DOTMP, MAG3 or DCTA. 35. The compound of claim 31, wherein said chelating group is DTPA. 36. The compound of claim 31, wherein the residue of the molecule comprising B-10 contains about 1 to 20 boron atoms. 37. The compound of claim 31, wherein the molecule comprising B-10 is a carbohydrate, amino acid, nucleoside or carborane. 38. The molecule containing B-10 is o-nido-carborane, m-nide-
32. The compound of claim 31, wherein the compound is carborane or p-nido-carborane. 39. The method according to claim 31, wherein the molecule comprising B-10 is o-carborane.
The compound according to the above. 40. The compound of claim 31, wherein the residue of the molecule comprising B-10 is attached to the 6-position of the compound of formula I or to the residue of a b-, d- or e-carboxamide group. 41. The compound of claim 31, wherein the residue of the compound of formula I is linked to the residue of a molecule comprising B-10 by a linker. 42. The compound of claim 41, wherein said linker comprises a non-metallic radionuclide. 43. The compound of claim 41, wherein the linker is between about 5 Angstroms and 50 Angstroms. 44. The compound of claim 1, further comprising a detectable radionuclide. 45. The compound of claim 2, wherein the detectable radionuclide is a non-metallic radionuclide. 46. The non-metallic radionuclide is carbon-11, fluorine-18, bromine-7.
6. The compound according to claim 3, which is iodine-123 or iodine-124. 47. The compound of claim 2, wherein the detectable radionuclide is directly attached to the compound of formula I. 48. The compound of claim 2, wherein said detectable radionuclide is linked to said compound of formula I by a linker. 49. The linker is of formula WA, wherein A is (C ₁ -C ₆ ) alkyl, (C ₂ -C ₆ ) alkenyl, (C ₂ -C ₆ ) alkynyl, (C ₃ -C _{6 8} )
Cycloalkyl or _(C 6 _{-C 10)} aryl, wherein W is -N (R)
C (= O)-, -C (= O) N (R)-, -OC (= O)-, -C (= O) O-
, -O-, -S-, -S (O)-, -S (O) _2- , -N (R)-, -C (= O
) - or a direct bond; wherein each R is H or (C 1 _{-C 6)} alkyl independently; A is optionally substituted by one or more radionuclides A compound according to claim 6. 50. The compound of claim 48, wherein the linker is between about 5 Angstroms and 50 Angstroms. 51. The linker of claim 48, wherein the linker is a divalent peptide or amino acid.
The compound according to the above. 52. A method according to claim 52, wherein the linker is poly-L-glutamic acid, poly-L-aspartic acid, poly-L-histidine, poly-L-ornithine, poly-L-serine, poly-L-threonine, poly-L-tyrosine, 49. The compound of claim 48, which is poly-L-lysine-L-phenylalanine or poly-L-lysine-L-tyrosine. 53. The method according to claim 53, wherein the linker is attached at the 6 position of the compound of formula I.
49. The compound of claim 48, wherein the compound is attached to the residue of an a-, b-, d-, or e-carboxamide group of the compound of 54. A compound wherein the residue of the compound of Formula I (FIG. 1) is 1) conjugated to a detectable radionuclide and 2) conjugated to a group comprising Gd-157. Acceptable salts thereof. 55. The group comprising Gd-157 has the formula QLW-Det,
Is CN, OH, CH _3, adenosyl, molecules containing B-10 or Q-L-W-,
Det, where Det is a chelating group containing Gd-157, L is or is not a linker, and W and Q are each independently -N (R) C (= O)
-, -C (= O) N (R)-, -OC (= O)-, -C (= O) O-, -O-,
—S—, —S (O) —, —S (O) ₂ —, —C (＝ O) —, —N (R) — or a direct bond, and each R is independently H or (C ₁ — C ₆₎ alkyl, a compound according to claim 54. 56. The compound of claim 54, wherein said detectable radionuclide is a non-metallic radionuclide. 57. The non-metallic radionuclide is carbon-11, fluorine-18, bromine-7.
57. The compound of claim 56 which is iodine-123 or iodine-124. 58. The compound of claim 54, wherein the detectable radionuclide is directly attached to the compound of formula I. 59. The compound of claim 54, wherein the detectable radionuclide is linked to the compound of formula I by a linker. 60. The linker is of formula WA wherein A is (C ₁ -C ₆ ) alkyl, (C ₂ -C ₆ ) alkenyl, (C ₂ -C ₆ ) alkynyl, (C ₃ -C _{6 8} )
Cycloalkyl or _(C 6 _{-C 10)} aryl, wherein W is -N (R)
C (= O)-, -C (= O) N (R)-, -OC (= O)-, -C (= O) O-
, -O-, -S-, -S (O)-, -S (O) _2- , -N (R)-, -C (= O
) - or a direct bond, each R is H or (C 1 _{-C 6)} alkyl independently, A is substituted by one or more radionuclides A compound according to claim 59. 61. The compound of claim 59, wherein the linker is between about 5 Angstroms and 50 Angstroms. 62. The linker of claim 59, wherein the linker is a divalent peptide or amino acid.
The compound according to the above. 63. A poly-L-glutamic acid, a poly-L-aspartic acid, a poly-L-histidine, a poly-L-ornithine, a poly-L-serine, a poly-L-threonine, a poly-L-tyrosine, 60. The compound of claim 59 which is poly-L-lysine-L-phenylalanine or poly-L-lysine-L-tyrosine. 64. The linker is attached to position 6 of the compound of formula I or to the residue of an a-, b-, d- or e-carboxamide group of the compound of formula I,
60. The compound of claim 59. 65. The compound of formula I (FIG. 1) wherein the residue is 1) a molecule comprising Gd-157 or a chelating group comprising Gd-157 and 2) one or more of the formula QLW-Det , Det is each independently a chelating group containing a metal radionuclide, L is each independently a linker or not, W and Q are each independently -N ( R) C (= O)-, -C (= O) N (R)-,
-OC (= O)-, -C (= O) O-, -O-, -S-, -S (O)-, -S (
_{O) 2 -, - C (} = O) -, - N (R) - or a direct bond, compounds each R is H or _(C 1 -C ₆₎ alkyl independently, or a pharmaceutically acceptable Be that salt. 66. The compound of claim 1 or 44, wherein the residue of the compound of formula I is also attached to a group comprising Gd-157. 67. A group comprising Gd-157 is of the formula QLW-Det;
Is CN, OH, CH _3, adenosyl, molecules containing B-10 or Q-L-W-,
Det, wherein Det is a chelating group containing Gd-157, L is or is not a linker, and W and Q are each independently -N (R) C (= O)-,-
C (= O) N (R)-, -OC (= O)-, -C (= O) O-, -O-, -S-
_{, -S (O) -, -} S (O) 2 -, - C (= O) -, - N (R) - or a direct bond, each R is independently H or _(C 1 _-C 6) 67. The compound of claim 66, which is an alkyl. 68. A pharmaceutical compound comprising the compound according to any one of claims 1 to 67 and a pharmaceutically acceptable carrier. 69. administering to a mammal an effective amount of a compound according to any of claims 1 to 67 in combination with a pharmaceutically acceptable vehicle, and providing neutron capture therapy. A method of treating a tumor in a mammal in need of such treatment. 70. Imaging a tumor in a mammal, comprising administering to the mammal a detectable amount of a compound according to any of claims 1 to 67 and detecting the presence of the compound. Method. 71. The method of claim 70, further comprising treating the tumor with neutron capture therapy. 72. A compound according to any one of claims 1 to 67 for use in medical treatment or diagnosis. 73. Use of a compound according to any of claims 1 to 67 for the manufacture of a medicament for imaging a tumor in a mammal. 74. Use of a compound according to any of claims 1 to 67 for the manufacture of a medicament for treating a tumor in a mammal in need of such treatment.