Thiohydantoin androgen receptor antagonists for the treatment of cancer
A technology of sulfo-receptor, which is applied in the direction of antineoplastic drugs, organic active ingredients, medical preparations containing active ingredients, etc., can solve the problem of tumor growth without compounds
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[0374]The preparation of compounds of the present invention can be found in U.S. Patent Application No. 13 / 579,009, filed February 16, 2011, entitled "Androgen Receptor Modulators and Uses Thereof," which claims the February 2010 The benefit of U.S. Provisional Patent Application No. 61 / 305,082 filed on the 16th, which is hereby incorporated by reference.
[0375] biological example
[0376] As used herein, the term "biological sample" includes, but is not limited to, cell cultures or extracts thereof; biopsy material obtained from mammals or extracts thereof; and blood, saliva, urine, feces, semen, tears, or other bodily fluids or its extracts.
[0377] Antagonism of receptors in biological samples can be used for various purposes known to those skilled in the art. Examples of such purposes include, but are not limited to, bioassays, gene expression studies, and biological target identification.
[0378] Certain embodiments of the invention relate to methods of treatment...
Embodiment 1
[0385] Compounds bind to radioligands for AR, GR, and ER
[0386] Radioligand binding assays were performed using cell extracts and ligands as described below. Complete methods are included in the cited publications. K d Values were determined by non-specific incubation assays.
[0387] receptor
[0388] GR (human) (agonist radioligand) IM-9 cells (cytosol)
[0389] [ 3 H] Dexamethasone 1.5nM 1.5nM triamcinolone acetonide (10 μM) 6h 4°C scintillation counting (Clark, A.F et al., (1996) Invest. Ophtalmol. Vis. Sci., 37:805-813).
[0390] ER (non-selective) (human) (agonist radioligand) MCF-7 cells (cytosol)
[0391] [ 3 H] Estradiol 0.4nM 0.2nM 17-β-estradiol (6μM) 20h 4°C scintillation counting (Parker, G.J et al. (2000) J.Biomol.Screen., 5:77-88).
[0392] AR (human) (agonist radioligand) LNCaP cells (cytosol)
[0393] [ 3 H] Methyltrienone 1nM 0.8nM Mibolone (1μM) for 24 hours, scintillation counting at 4°C.
[0394] Zava, D.T et al. (1979) Endocrinology, 10...
Embodiment 2
[0406] Antagonism of AR (WT or F876L) reporter assays
[0407] LNCaP AR(cs) and LNCaP F876L luciferase cell lines were generated by transduction of each cell line (Description of cell line generation Joseph JD, Lu N, Qian J, Sensintaffar J, Shao G, Brigham D, Moon M, ManevalEC, Chen I, Darimont B, Hager JH. Clinically relevant androgen receptor mutations confer resistance to second-generation antiandrogens enzalutamide and ARN-509. Discovering Tumors (Cancer Discov 2013;3:1020-1029) , which has an androgen response element firefly luciferase lentiviral construct with an MOI (multiplicity of infection) of 50, following the manufacturer's instructions (Qiagen). Stable mixed population cell lines were generated using puromycin (Life Technologies) selection at 1:10,000 v / v. The following protocol was used for both cell lines and for testing compounds of formula (I) of the present invention.
[0408] LNCaP cells were grown to approximately 80% confluency, media removed and cell...
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