NOVEL INHIBITORS OF 17?-HSD7 AND USES THEREOF
20240287127 ยท 2024-08-29
Assignee
Inventors
- Sheng-Xiang LIN (Qu?bec, CA)
- Donald Poirier (L'Ancienne-Lorette, CA)
- Ren? MALTAIS (Qu?bec, CA)
- Jean-Yves SANC?AU (Qu?bec, CA)
Cpc classification
A61K45/06
HUMAN NECESSITIES
A61K31/58
HUMAN NECESSITIES
International classification
C07J73/00
CHEMISTRY; METALLURGY
A61K31/58
HUMAN NECESSITIES
A61K45/06
HUMAN NECESSITIES
Abstract
Novel chemical agents are described herein. More specifically, a novel inhibitor of 17?-HSD7 for decreasing estradiol concentrations while restoring dihydrotestosterone (DHT) concentrations in breast cancer cells is disclosed herein. In a particular embodiment, the inhibitor of 17?-HSD7 has the following structure: (Formula I) A process for producing the novel inhibitors of 17?-HSD7 and their use in the manufacture of pharmaceutical formulations and/or combinations is also disclosed.
##STR00001##
Claims
1. An inhibitor of 17?-HSD7, wherein the inhibitor has the structure of Formula I: ##STR00009## wherein R is alkyl.sub.(C?12) or cycloalkyl.sub.(C?12); and X is O or S; or a pharmaceutically acceptable salt, prodrug or solvate thereof.
2. The inhibitor of claim 1, having the structure: ##STR00010##
3. A method of treating cancer in a subject, the method comprising administering to the subject the inhibitor of claim 1 or 2.
4. The method of claim 3, wherein the cancer is an estrogen-sensitive cancer.
5. The method of claim 4, wherein the cancer is breast cancer.
6. The method of claim 5, wherein the inhibitor is the inhibitor of claim 2.
7. The method of claim 3, further comprising treating the subject with a secondary cancer therapy.
8. The method of claim 6, further comprising treating the subject with a secondary cancer therapy.
9. The method of claim 7 or 8, wherein the secondary cancer therapy is selected from the group consisting of chemotherapy, toxin therapy, radiation therapy, hormone or anti-hormone therapy, surgery, cryotherapy, or immunotherapy.
10. The method of any of claims 3 to 9, further comprising administering the inhibitor at least a second time.
11. The method of any of claims 3 to 9, wherein the inhibitor is administered intravenously, intra-arterially, subcutaneously, topically, or intramuscularly.
12. The method of any of claims 3 to 9, wherein the inhibitor is administered systemically, regionally to a tumor/disease site, locally to a tumor/disease site, into tumor/tissue vasculature or intratumorally.
13. The method of any of claims 3 to 12, wherein the cancer is multi drug resistant.
14. The method of any of claims 3 to 12, wherein the cancer is metastatic.
15. The method of any of claims 3 to 12, wherein the cancer is recurrent.
16. The method of any of claims 3 to 12, wherein treating comprises inhibiting cancer growth, killing cancer cells, reducing tumor burden, reducing tumor size, improving said subject's quality of life or prolonging said subject's length of life.
17. The method of any of claims 3 to 16, wherein the subject is a human.
18. The method of any of claims 3 to 16, wherein the subject is a non-human animal.
19. A pharmaceutical composition comprising a pharmaceutically acceptable amount of the inhibitor according to claim 1 or 2 and a pharmaceutically acceptable carrier.
20. Use of the inhibitor according to claim 1 or 2 for the prophylaxis or treatment of a disease.
21. The use of claim 20, wherein the disease is associated with uncontrolled cell growth, proliferation and/or survival.
22. The use of claim 21, wherein the disease comprises breast cancer.
23. Use of the pharmaceutical composition of claim 19 for the prophylaxis or treatment of a disease.
24. The use of claim 23, wherein the disease is associated with uncontrolled cell growth, proliferation and/or survival.
25. The use of claim 24, wherein the disease comprises breast cancer.
Description
BRIEF DESCRIPTION OF THE DRAWINGS/FIGURES
[0054] The following figures/drawings form part of the present specification and are included to further demonstrate certain aspects of the present specification. The present specification may be better understood by reference to one or more of these figures/drawings in combination with the detailed description. In the appended drawings/figures:
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DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS
[0066] The present disclosure relates to novel inhibitors of 17?-HSD7. More specifically, but not exclusively, the present disclosure relates to novel inhibitors of 17?-HSD7 capable of decreasing estradiol concentrations while restoring dihydrotestosterone (DHT) concentrations in breast cancer cells. The present disclosure also relates to the synthesis of inhibitors of 17?-HSD7. Moreover, the present disclosure relates to compositions and pharmaceutical formulations comprising an inhibitor of 17?-HSD7. Yet moreover, the present disclosure relates to compositions and pharmaceutical formulations comprising an inhibitor of 17?-HSD7 for decreasing estradiol concentrations, while restoring dihydrotestosterone (DHT) concentrations in breast cancer cells. Furthermore, the present disclosure relates to methods of treatment comprising the use of an inhibitor of 17?-HSD7. Yet furthermore, the present disclosure relates to methods of treatment comprising the use of an inhibitor of 17?-HSD7 for decreasing estradiol concentrations, while restoring dihydrotestosterone (DHT) concentrations in breast cancer cells.
[0067] In a particular aspect, the present disclosure relates to an inhibitor of 17?-HSD7 having the structure of Formula I:
##STR00006##
[0068] wherein R is alkyl.sub.(C?12) or cycloalkyl.sub.(C?12); and X is O or S; or a pharmaceutically acceptable salt, prodrug or solvate thereof.
[0069] In a further particular aspect, the present disclosure relates to an inhibitor of 17?-HSD7 having the structure:
##STR00007##
[0070] These and other aspects of the disclosure are described in greater detail below.
[0071] The 17?-HSD7 inhibitors of the present disclosure are shown, for example, above in the summary section and in the examples and claims below. INH7-464 can be synthesized according to the methods described, for example, in the Examples section below.
[0072] Atoms making up INH7-464 and other 17?-HSD7 inhibitors of the present disclosure are intended to include all isotopic forms of such atoms. Isotopes, as used herein, include those atoms having the same atomic number but different mass numbers. By way of general example and without limitation, isotopes of hydrogen include tritium and deuterium, and isotopes of carbon include .sup.13C and .sup.14C.
[0073] INH7-464, and other 17?-HSD7 inhibitors of the present disclosure may also exist in prodrug form. Since prodrugs are known to enhance numerous desirable qualities of pharmaceuticals (e.g., solubility, bioavailability, manufacturing, etc.), INH7-464 may, if desired, be delivered in prodrug form. Thus, the disclosure contemplates prodrugs of the 17?-HSD7 inhibitors of the present disclosure. Prodrugs of INH7-464, and other 17?-HSD7 inhibitors of the present disclosure may be prepared by modifying functional groups present in the compound in such a way that the modifications are cleaved, either in routine manipulation or in vivo, to the parent compound. Accordingly, prodrugs include, for example, compounds described herein in which an amino, or carboxy group is bonded to any group that, when the prodrug is administered to a subject, cleaves to form an amino, or carboxy, respectively.
[0074] It should be recognized that the particular anion or cation forming a part of any salt form of INH7-464, or any other 17?-HSD7 inhibitor of the present disclosure is not critical, so long as the salt, as a whole, is pharmacologically acceptable. Additional examples of pharmaceutically acceptable salts and their methods of preparation and use are presented in Handbook of Pharmaceutical Salts: Properties, and Use (2002), which is incorporated herein by reference.
[0075] Those skilled in the art of organic chemistry will appreciate that many organic compounds can form complexes with solvents in which they are reacted or from which they are precipitated or crystallized. These complexes are known as solvates. For example, a complex with water is known as a hydrate. Solvates of INH7-464, or any other 17?-HSD7 inhibitor of the present disclosure are within the scope of the present disclosure. It will also be appreciated by those skilled in organic chemistry that many organic compounds can exist in more than one crystalline form. For example, crystalline forms may vary from solvate to solvate. Thus, all crystalline forms of INH7-464, or any other 17?-HSD7 inhibitor of the present disclosure or the pharmaceutically acceptable solvates thereof are within the scope of the present disclosure.
Synthetic Methods
[0076] In some aspects, INH7-464, or any other 17?-HSD7 inhibitor of the present disclosure can be synthesized using the methods of organic chemistry as described in this application. These methods can be further modified and optimized using the principles and techniques of organic chemistry as applied by a person skilled in the art. Such principles and techniques are taught, for example, in March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure (2007), which is incorporated by reference herein.
Process Scale-Up
[0077] The synthetic methods described herein can be further modified and optimized for preparative, pilot- or large-scale production, using the principles and techniques of process chemistry as applied by a person skilled in the art. Such principles and techniques are taught, for example, in Practical Process Research & Development (2000), which is incorporated by reference herein. The synthetic method described herein may be used to produce preparative scale amounts of INH7-464, or any other 17?-HSD7 inhibitor of the present disclosure.
Chemical Definitions
[0078] When used in the context of a chemical group: hydrogen means H; hydroxy means OH; oxo means ?O; carbonyl means C(?O); carboxy means C(?O)OH (also written as COOH or CO.sub.2H); halo means independently F, Cl, Br or I; amino means NH.sub.2; hydroxyamino means NHOH; nitro means NO.sub.2; imino means ?NH; cyano means CN; isocyanate means N?C?O; azido means N.sub.3; in a monovalent context phosphate means OP(O)(OH).sub.2 or a deprotonated form thereof; in a divalent context phosphate means OP(O)(OH)O or a deprotonated form thereof; mercapto means SH; and thio means ?S; sulfato means SO.sub.3H, sulfamido means S(O).sub.2NH.sub.2, sulfonyl means S(O).sub.2; and sulfinyl means S(O).
[0079] In the context of chemical formulas, the symbol means a single bond,
means a double bond, and
means a triple bond. The symbol
represents an optional bond, which if present is either single or double. The symbol
represents a single bond or a double bond. Any undefined valency on an atom of a structure shown in this application implicitly represents a hydrogen atom bonded to that atom.
[0080] The term alkyl as used herein, represents a monovalent group derived from a straight or branched chain saturated hydrocarbon comprising, unless otherwise specified, from 1 to 12 carbon atoms and is exemplified by methyl, ethyl, n- and iso-propyl, n-, sec-, iso- and tert-butyl, neopentyl, nonyl and the like and may be optionally substituted with one, two, three or more substituents.
[0081] The term cycloalkyl as used herein, represents a monovalent saturated or unsaturated non-aromatic cyclic hydrocarbon group of three to 12 carbon atoms, unless otherwise specified, and is exemplified by cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, bicyclo[2.2.1.]heptyl and the like. The cycloalkyl groups of the present disclosure can be optionally substituted.
[0082] An alkane or cycloalkane refers to the compound H-R, wherein R is alkyl or cycloalkyl as these terms are defined above. When any of these terms is used with the substituted modifier one or more hydrogen atoms have been independently replaced by a group, non-limiting examples of which include. OH, F, Cl, Br, I, NH.sub.2, NO.sub.2, CO.sub.2H, CO.sub.2CH.sub.3, CN, SH, OCH.sub.3, OCH.sub.2CH.sub.3, C(O)CH.sub.3, NHCH.sub.3, NHCH.sub.2CH.sub.3, N(CH.sub.3).sub.2, C(O)NH.sub.2, OC(O)CH.sub.3, or S(O).sub.2NH.sub.2. The following groups are non-limiting examples of substituted alkyl groups: CH.sub.2OH, CH.sub.2Cl, CF.sub.3, CH.sub.2CN, CH.sub.2C(O)OH, CH.sub.2C(O)OCH.sub.3, CH.sub.2C(O)NH.sub.2, CH.sub.2C(O)CH.sub.3, CH.sub.2OCH.sub.3, CH.sub.2OC(O)CH.sub.3, CH.sub.2NH.sub.2, CH.sub.2N(CH.sub.3).sub.2, and CH.sub.2CH.sub.2Cl.
EXAMPLES
[0083] The following examples are included to demonstrate preferred embodiments of the disclosure. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow represent techniques discovered by the inventors to function well in the practice of the invention, and thus can be considered to constitute preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention.
Example 1Synthesis of INH7-464
[0084] ##STR00008##
[0085] (4aR,4bS,6aS,7R,9aS,9bR,11aR)-4a,6a-dimethyl-3-nonyltetradecahydrospiro [indeno[5,4-f]quinoline-7,5-oxazolidine]-2,2(1H)-dione (INH 464): To an ice cooled solution of N-nonylamino-4-azasteroid (84 mg, 0.18 mmol) in pyridine (5 ml) was added DMAP (2 mg) and triphosgene (80 mg, 0.27 mL). The cooling bath was subsequently removed and the resulting mixture was stirred at rt for 10 h. The reaction mixture was then poured into ice and extracted with DCM. The organic extracts were washed with brine and dried with sodium sulfate. After evaporation, the crude compound was purified by flash chromatography with DCM/MeOH (99:1) to yield 60 mg (69% yield) of INH 464 as a yellowish solid. .sup.1HNMR (400 MHz, CDCl.sub.3) ?=5.39 (s, 1H, NH), 3.48 (d, J=9.2 Hz, 1H, NCH.sub.2), 3.25 (d, J=9.2 Hz, 1H, NCH.sub.2), 3.22 (t, J=7.4 Hz, 2H, NCH.sub.2CH.sub.2), 3.06 (dd, J=4.1 and 11.8 Hz, 1H, H.sub.5?), 2.47-2.33 (m, 2H), 2.18-2.11 (m, 1H), 1.96-1.22 (m, 26H), 1.06 (m, 1H), 0.90 (s, 3H), 0.88 (s, 3H), 0.87 (t, J=7.0 Hz, 3H, CH.sub.2CH.sub.3), 0.72 (s, 3H). MS (APCI positive) m/z [M+H] 473.8. HPLC purity=95.3%.
Comparison of the Effect of Various INH7 Inhibitors on Cancer Cell Proliferation
[0086] The effects of various inhibitors, including INH7-464, on MCF-7 cell proliferation over a period of 144 hours is illustrated in
[0087] The effect of INH7-10 and INH7-464 on both xenograft breast tumor cell models MCF-7 and T47D was further investigated. For the MCF-7 xenograft model, a decrease in the tumor size of more than 60%, relative to the control (E1), could be observed with both INH7-10 (59%?5%) and INH7-464 (60%?5%) on day 22. For the T47D xenograft model, a decrease in the tumor size of more than 51%, relative to the control (E1), could be observed with INH7-464 (51%?5%) on day 22. A decrease in the tumor size of more than 31%, relative to the control (E1), could be observed with INH7-10 (31%?6%) on day 22 for the T47D xenograft model. INH7-464, thus demonstrates significant efficacy in reducing xenograft tumors induced by both MCF-7 and T47D breast cancer cell lines.
Toxicity Assay for INH7-464
[0088] Paclitaxel (PTX) was used as a reference compound and was dissolved in DMSO for the stock solutions. The stock solutions were subsequently diluted at multiple concentrations with culture media in order to obtain the final desired concentrations ranging from 1 nM-100 ?M. The incubation time was 3 days. The IC.sub.50 values (at which 50% of cell growth inhibitionconcentration that inhibits 50% of cell proliferationis observed) were calculated using an iterative least square regression method. The cytotoxicity values for the INH7-464 inhibitor were lower than those for Paclitaxel (PTX) in the different cell lines tested [581.6 nM (MCF-7); and 2669 nM (MCF-10A) respectively](cf. Table 3).
[0089] The toxicity of INH7-464 was subsequently evaluated in mice (8 mice/group) at various concentrations and was found to be very well tolerated after weekly subcutaneous injection at various concentrations (20-60 mg/kg) over a period of 21 days; there being no apparent toxicity at 60 mg/day/mouse (s.c.) (
General Methods and Materials
[0090] Reagents and solvents were obtained from commercial suppliers (Sigma Aldrich, Strem, Combi-blocks, Alfa Aesar) and used without further purification, unless otherwise noted. All reactions that were moisture and air-sensitive were carried out in flame-dried glassware, under an argon atmosphere. Reaction progress was monitored by thin layer chromatography (TLC), using EMD silica gel 60 F254 aluminum plates. Spots were visualized with UV light (254 nm), followed by staining using a cerium ammonium molybdate (CAM) solution or a potassium-permanganate solution, followed by heating on a hot plate. SiliCycle? R10030B 230-400 mesh silica gel (Qu?bec, QC, Canada) was used for flash chromatography. High-performance liquid chromatography (HPLC) analyses for chemical purities were performed on a Shimadzu Prominence instrument (Kyoto, Japan) using a diode array detector and an Altima C18 analytical reverse phase column. Nuclear magnetic resonance (NMR) spectra were recorded on a Bruker Avance 400 digital spectrometer (Billerica, MA, USA) at 400 MHz for .sup.1H NMR. The following abbreviations were used to designate multiplicities: s=singlet, d=doublet, t=triplet, q=quartet, quint=quintuplet, m=multiplet, br=broad. Low-resolution mass spectra (LRMS) were recorded on a Shimadzu Prominence instrument (Kyoto, Japan) equipped with a Shimadzu LCMS-2020 mass spectrometer and an APCI (atmospheric pressure chemical ionization) probe.
[0091] INH7-464 was administered by s.c. injection. INH7-464 was first dissolved in DMSO, followed by the addition of 0.4% methylcellulose to obtain a final 8% concentration of co-solvent (methylcellulose/DMSO (92/8). The injected volume was 0.1 mL (s.c.). The protocol was applied to 2 groups (8 mice/group), and divided-up as follows: Control Group: vehicle administrated weekly by s.c. injection of 0.4% methylcellulose/DMSO (92:8) for 3 weeks; INH7-464 Group: INH7-464 administrated weekly by s.c. injection (20, 30 and 60 mg/kg) for 3 consecutive weeks. All the mice were weighed every day. The liver, kidney, ovaries, and uterus were collected and weighed following 3 weeks of treatment.
[0092] Female athymic nude mice (4 weeks) were ovariectomized under isoflurane-induced anesthesia. One week after ovariectomy (OVX), 5?10.sup.6 MCF-7 cells or 10?10.sup.6 T47D cells suspended in a 100 ml mixture of Matrigel (BD)/culture medium (50:50) were administered by s.c. injection on both side of each mice. All mice were supplemented by daily s.c. injection 0.1 ?g Estradiol (E2) for 5 weeks. Group 1Control: E1 0.1 ?g/mice/day; Group 2: E2 0.1 ?g/mice/day; Group 3: OVX (Group of mice having had an ovariectomy); Group 4: E1 0.1 ?g/mice/day; INH7(10) 5 mg/kg/day; Group 5: E1 0.1 ?g/mice/day; INH7(464) 5 mg/kg/day. The animals were euthanized on day 23, followed by the collection of tumor specimens and blood samples. The estrogen-sensitive organs (uterus and vagina), liver and kidney were removed and excised of fat and weighed. Tumor specimens were photographed and weighed. E1: Estrone; E2: Estradiol; OVX: group of mice having had an ovariectomynot treated with an INH7 inhibitor, but instead were subjected to daily subcutaneous (s.c.) injection with vehicle (0.4% methylcellulose/DMSO (92/8)).
[0093] All of the compounds and/or methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compounds and methods of this disclosure have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the compounds and/or methods and in the steps or in the sequence of steps of the methods described herein without departing from the concept, spirit and scope of the disclosure. More specifically, it will be apparent that certain agents which are chemically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the disclosure as defined by the appended claims.
TABLE-US-00001 TABLE 1 Effect of various inhibitors of 17b-HSD1 (INH7-10 and INH7- 464) on the growth of human MCF-7 breast tumors in nude mice over a period of 22 days expressed as % of initial growth. Days E1 E2 OVX INH7 (10) INH7 (464) 0 100 100 100 100 100 5 102.886 82.8535 73.9924 95.5496 88.1406 9 100.227 97.2662 73.9721 78.5457 81.8741 12 109.033 131.541 83.8482 85.0172 84.2617 13 95.3089 123.252 69.3147 80.2823 66.5956 19 100.249 108.413 67.2073 71.1033 67.8843 22 102.648 120.974 48.2038 59.4114 59.6876
TABLE-US-00002 TABLE 2 Effect of various inhibitors of 17b-HSD1 (INH7-10 and INH7- 464) on the growth of human T47D breast tumors in nude mice over a period of 22 days expressed as % of initial growth. Days E1 E2 OVX INH7 (10) INH7 (464) 0 100 100 100 100 100 5 93.64 112.55 105.67 89.41 100.51 9 113.43 120.84 81.8141 98.79362 78.1158 12 112.0519 121.306 78.6751 80.3944 76.9139 15 122.0034 150.417 69.5895 87.12289 63.5165 19 112.9356 144.117 68.5695 80.2485 67.9195 22 112.831 138.919 67.985 75.3021 50.9558
TABLE-US-00003 TABLE 3 Cytotoxicity of the INH7-464 inhibitor on the MCF-7 and MCF-10A cell lines. Cytotoxicity Paclitaxel (PTX) INH7 (464) IC.sub.50 (MCF-7) 3.082 nM 581.6 nM IC.sub.50 (MCF-10A) 0.6373 nM 2669 nM
REFERENCES
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