Dual cyp17a1-hdac inhibitors

Abstract

The present disclosure relates to first-in-class dual CYP17A1-HDAC inhibitors, process of preparing the same and uses thereof in treatment of diseases / disorders mediated by CYP17A1, HDAC and / or both, in particular cancer (e.g., glioblastoma or prostate cancer).

Description

DUAL CYP17A1-HDAC INHIBITORS PRIORITY INFORMATION

[0001] This application claims benefit of and priority to U.S. Provisional Patent Application No. 63 / 518,489, filed August 9, 2023, the contents of which is incorporated by reference in its entirety. FILED OF THE INVENTION

[0002] The present disclosure relates to a field of pharmaceuticals. Particularly, the present disclosure pertains to first-in-class dual CYP17A1-HDAC inhibitors, a process of preparing the same and uses thereof in treatment of diseases / disorders mediated by CYP17A1, HDAC and / or both. BACKGROUND OF THE INVENTION

[0003] Treatments of cancer are important and demands thereof are still increasing nowadays. Among various types of cancer, glioblastoma (GBM) is a highly complex, aggressive, and treatment-resistant brain tumor characterized by a genetically unstable and highly infiltrative population of cells. GBM can develop in any area of the brain, most frequently so in the cerebral hemispheres, which are its largest and most intricate structures. The multimodality treatment protocol for GBM comprises surgical resection flanked by radiotherapy and chemotherapy. Disappointingly, this trimodal therapy in many cases is unable to garner satisfactory response as the cytological heterogeneity of GBM makes the complete surgical removal of the tumor a Gordian knot. The challenges arising due to incomplete surgical resection are further complicated by i) a lack of potential chemotherapeutic options with temozolomide (oral alkylating agent) as the only available first-line agent ii) hindrance in the clinical utility of temozolomide owing to methylguanine DNA methyltransferase mediated innate resistance iii) Tumor recurrence due to the existence of a highly tumorigenic population of cells known as glioma stem cells (GSCs). Given the unsatisfactory outcomes of the GBM treatment regime, it is imperative to expand the armoryof potential chemotherapeutics (anti-GBM agents) in pursuit of an effective treatment protocol. SUMMARY OF THE INVENTION

[0004] The present disclosure provides dual CYP17A1-HDAC inhibitors for preventing and / or treating diseases / disorders mediated by CYP17A1, HDAC and / or both (such as cancer, e.g., glioblastoma or prostate cancer).

[0005] The present disclosure provides a compound of Formula (I):or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, wherein: X is O, NRaor S wherein Rais H or C1-5alkyl, preferably X is O or NH; R is H, C1-5alkyl, -C(=O)Rb, -C(=O)ORbor –C(=O)-NRbwherein Rbis H or C1-5alkyl, preferably R is H; and n is an integer ranging from 2 to 10, preferably 4 to 9, more preferably 5 to 8; wherein each of RA1, RA2, RA3 and RA4 is independently halogen, hydroxy, C1-5alkyl or haloC1-5alkyl; q, r, s and t each independently represent an integer ranging from 0 to 3, provided that the sum of q, r, s and t ranges from 0 to 5.

[0006] The subject disclosure also provides a compound of formula (IA):(IA), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, wherein: X is O, NRaor S wherein Rais H or C1-5alkyl, preferably X is O or NH; R is H, C1-5alkyl, -C(=O)Rb, -C(=O)ORbor –C(=O)-NRbwherein Rbis H or C1-5alkyl, preferably R is H; and n is an integer ranging from 2 to 10, preferably 4 to 9, more preferably 5 to 8; wherein each of RA1, RA2, RA3 and RA4 is independently halogen, hydroxy, C1-5alkyl or haloC1-5alkyl; q, r, s and t each independently represent an integer ranging from 0 to 3, provided that the sum of q, r, s and t ranges from 0 to 5.

[0007] The subject disclosure also provides a compound of formula (IB):or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, wherein: X is O, NRaor S wherein Rais H or C1-5alkyl, preferably X is O or NH;R is H, C1-5alkyl, -C(=O)Rb, -C(=O)ORbor –C(=O)-NRbwherein Rbis H or C1-5alkyl, preferably R is H; and n is an integer ranging from 2 to 10, preferably 4 to 9, more preferably 5 to 8; wherein each of RA1, RA2, RA3 and RA4 is independently halogen, hydroxy, C1-5alkyl or haloC1-5alkyl; q, r, s and t each independently represent an integer ranging from 0 to 3, provided that the sum of q, r, s and t ranges from 0 to 5.

[0008] In one embodiment, the compound of Formula (I) is Formula (I'):or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, wherein: R is H, C1-5alkyl, -C(=O)Rb, -C(=O)ORbor –C(=O)-NRbwherein Rbis H or C1-5alkyl, preferably R is H; and n is an integer ranging from 2 to 10, preferably 4 to 9, more preferably 5 to 8; wherein each of RA1, RA2, RA3and RA4is independently halogen, hydroxy, C1-5alkyl or haloC1-5alkyl; q, r, s and t each independently represent an integer ranging from 0 to 3, provided that the sum of q, r, s and t ranges from 0 to 5.

[0009] In one embodiment, the compound of Formula (I) is Formula (IA'):(IA'), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, wherein: R is H, C1-5alkyl, -C(=O)Rb, -C(=O)ORbor –C(=O)-NRbwherein Rbis H or C1-5alkyl, preferably R is H; and n is an integer ranging from 2 to 10, preferably 4 to 9, more preferably 5 to 8; wherein each of RA1, RA2, RA3 and RA4 is independently halogen, hydroxy, C1-5alkyl or haloC1-5alkyl; q, r, s and t each independently represent an integer ranging from 0 to 3, provided that the sum of q, r, s and t ranges from 0 to 5.

[0010] In one embodiment, the compound of Formula (I) is Formula (IB'):(IB'), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, wherein: R is H, C1-5alkyl, -C(=O)Rb, -C(=O)ORbor –C(=O)-NRbwherein R2is H or C1-5alkyl, preferably R is H; andn is an integer ranging from 2 to 10, preferably 4 to 9, more preferably 5 to 8; wherein each of RA1, RA2, RA3 and RA4 is independently halogen, hydroxy, C1-5alkyl or haloC1-5alkyl; q, r, s and t each independently represent an integer ranging from 0 to 3, provided that the sum of q, r, s and t ranges from 0 to 5.

[0011] In one embodiment, the compound of Formula (I) is Formula (I"):or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, wherein: R is H, C1-5alkyl, -C(=O)Rb, -C(=O)ORbor –C(=O)-NRbwherein Rbis H or C1-5alkyl, preferably R is H; and n is an integer ranging from 2 to 10, preferably 4 to 9, more preferably 5 to 8; wherein each of RA1, RA2, RA3 and RA4 is independently halogen, hydroxy, C1-5alkyl or haloC1-5alkyl; q, r, s and t each independently represent an integer ranging from 0 to 3, provided that the sum of q, r, s and t ranges from 0 to 5.

[0012] In one embodiment, the compound of Formula (I) is Formula (IA"):or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, wherein: R is H, C1-5alkyl, -C(=O)Rb, -C(=O)ORbor –C(=O)-NRbwherein Rbis H or C1-5alkyl, preferably R is H; and n is an integer ranging from 2 to 10, preferably 4 to 9, more preferably 5 to 8; wherein each of RA1, RA2, RA3 and RA4 is independently halogen, hydroxy, C1-5alkyl or haloC1-5alkyl; q, r, s and t each independently represent an integer ranging from 0 to 3, provided that the sum of q, r, s and t ranges from 0 to 5.

[0013] In one embodiment, the compound of Formula (I) is Formula (IB"):or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, wherein: R is H, C1-5alkyl, -C(=O)Rb, -C(=O)ORbor –C(=O)-NRbwherein Rbis H or C1-5alkyl, preferably R is H; andn is an integer ranging from 2 to 10, preferably 4 to 9, more preferably 5 to 8; wherein each of RA1, RA2, RA3 and RA4 is independently halogen, hydroxy, C1-5alkyl or haloC1-5alkyl; q, r, s and t each independently represent an integer ranging from 0 to 3, provided that the sum of q, r, s and t ranges from 0 to 5.

[0014] In any preceding embodiments of Formula (I), (IA) or (IB), X is O, NRaor S wherein Rais H or C1-5alkyl, R is H or C1-5alkyl, and n is an integer ranging from 5 to 8.

[0015] In any preceding embodiments of Formula (I), (IA) or (IB), X is O, NRaor S wherein Rais H or C1-5alkyl, R is -C(=O)Rb, -C(=O)ORbor –C(=O)-NRb, wherein Rbis H or C1-5alkyl, and n is an integer ranging from 5 to 8.

[0016] In any preceding embodiments of Formula (I), (IA) or (IB), X is O, NRaor S wherein Rais H or C1-5alkyl, R is H and n is an integer ranging from 4 to 9.

[0017] In any preceding embodiments of Formula (I), (IA) or (IB), X is O or NRawherein Rais H or C1-5alkyl, R is H or C1-5alkyl and n is an integer ranging from 5 to 8.

[0018] In any preceding embodiments of Formula (I), (IA) or (IB), X is O or NRawherein Rais H or C1-5alkyl, R is -C(=O)Rb, -C(=O)ORbor –C(=O)-NRb, wherein Rbis H or C1-5alkyl, and n is an integer ranging from 5 to 8.

[0019] In any preceding embodiments of Formula (I), (IA) or (IB), X is O or NRawherein Rais H or C1-5alkyl, R is H and n is an integer ranging from 4 to 9.

[0020] In any preceding embodiments, R is H and n is an integer ranging from 5 to 8.

[0021] In any preceding embodiments, each of RA1, RA2, RA3 and RA4 is independently halogen, hydroxy, C1-5alkyl or haloC1-5alkyl; q, r, s and t each independently represent an integer ranging from 0 to 1. In any preceding embodiments, each of RA1, RA2, RA3 and RA4 is independently halogen, hydroxy, C1-5alkyl or haloC1-5alkyl; q, r, s and t each independently represent an integer ranging from 0 to 1 and the sum of q, r, s and t ranges from 0 to 2.

[0022] In any preceding embodiments, the sum of q, r, s and t is 0.

[0023] Examples of the compounds of Formula (I) include, but are not limited to:or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof.

[0024] The subject disclosure also provides a process of preparing a compound of Formula (I), Formula (IA), Formula (IB), Formula (I'), Formula (IA'), Formula (IB'), Formula (I"), Formula (IA") or Formula (IB").

[0025] In one embodiment, the process uses a compound of Formula (II)as a reactant or intermediate to prepare the compound of Formula (I), Formula (IA), Formula (IB), Formula (I'), Formula (IA'), Formula (IB'), Formula (I"), Formula (IA") or Formula (IB"), wherein X is O, NRaor S, wherein Rais H or C1-5alkyl, and the definition of RA1, RA2, RA3, RA4, q, r, s and t are as those described above.

[0026] In one embodiment, the compound of Formula (II) is Compound (8) or (9).

[0027] In one embodiment, the compound of Formula (II) is Compound (10) or (11).

[0028] The subject application also provides a method of treating a disease or disorder mediated by CYP17A1, HDAC or both, comprising administering the compound describedherein or a pharmaceutically acceptable salt, hydrate, solvate or prodrug of any of the foregoing to a subject in need thereof.

[0029] The subject application also provides a pharmaceutical composition comprising the compound described herein or a pharmaceutically acceptable salt, hydrate, solvate or prodrug of any of the foregoing, and a pharmaceutically acceptable carrier.

[0030] The subject application also provides the compound described herein or a pharmaceutically acceptable salt, hydrate, solvate or prodrug of any of the foregoing for use in therapy, preferably in the treatment of a disease or disorder mediated by CYP17A1, HDAC or both.

[0031] In a further embodiment, the HDAC described herein is HDAC6.

[0032] The subject application also provides use / methods of the compound described herein or a pharmaceutically acceptable salt, hydrate, solvate or prodrug of any of the foregoing in the manufacture of a medicament for treating a cancer.

[0033] In a further embodiment, disease or disorder described herein is a CYP17A1-HDAC6 mediated disease or disorder. Preferably, the disease or disorder described herein is a cancer. In some embodiments, the cancers are hematological malignancies or solid tumors.

[0034] Examples of the cancer include, but are not limited to, lymphoma, myeloma (such as multiple myeloma), leukemia, melanoma, lung cancer (such as small cell lung cancer and non-small cell lung carcinoma (NSCLC) (such as squamous cell carcinoma, adenocarcinoma and large cell carcinoma)), breast cancer (such as metastatic breast cancer), cervical cancer, colorectal cancer (CRC), cholangiocarcinoma, esophageal cancer, liver cancer, colon cancer, pancreatic cancer, prostate cancer, stomach cancer, renal cancer and brain cancer.

[0035] In a further embodiment, the brain cancer is a primary brain tumor or prostate cancer or a metastatic brain cancer or prostate cancer. In one embodiment, the brain cancer is glioblastoma (GBM), medulloblastoma, or acoustic neuroma.BRIEF DESCRIPTIONS OF THE DRAWINGS

[0036] Fig. 1A shows the anti-GBM activity of compound 12 in suppressing cell viability in TMZ-resistant U87MG-R and A172-R cells.

[0037] Figs. 1B and 1C show docking of abiraterone (compound 8) and compound 12 with CP17A1 and docking of compound 12 and trichostatin A with HDAC6.

[0038] Figs. 2A-2E show the effect of Compound 12 on viability of normal mouse astrocyte, efficiency of gene manipulation and the effect of Compound 12 (C12) on cell cycle progression and subG1 accumulation.

[0039] Figs. 3A-3F show that Compound 12 induces DNA damage, increases ROS accumulation, decreases the levels of antioxidant proteins and enhances ROS production and induced ROS leakage from mitochondria.

[0040] Figs. 4A-4C show that Compound 12 suppresses the expression of recurrence-associated genes in GBM.

[0041] Figs. 5A-5H show the effect of compound 12 on the growth of TMZ-resistant Pt3R cells in vivo.

[0042] Figs. 6A and 6B show the effect of compound 12 on the growth of 22rv1 prostate cancer cells in vivo. DETAILED DESCRIPTION OF THE INVENTION

[0043] In order to facilitate understanding of the disclosure herein, terms as used herein are hereby defined below.

[0044] In the context of the specification and the claims, the singular forms "a", "an" and "the" include plural referents, unless specifically indicated otherwise. Unless otherwise stated, any and all examples or exemplary language (e.g., "such as") provided herein are merely used for better illustration of the present invention, instead of limiting the scope of the present invention.

[0045] It is to be understood that any numerical range recited in this specification is intended to include all sub-ranges encompassed therein. For example, a range from "50 to 70°C" includes all sub-ranges and specific values between the stated minimum value of 50°C and the stated maximum value of 70°C, inclusive, e.g. from 58°C to 67°C, and from 53°C to 62°C, 60°C or 68°C. Since the numerical ranges disclosed are continuous, they contain each numerical value between the minimum and maximum value. Unless otherwise specified, the various numerical ranges indicated in this specification are approximate.

[0046] In the present invention, the term "about" refers to an acceptable deviation of a given value measured by a person of ordinary skill in the art, depending, in part, on how to measure or determine the value.

[0047] In the present invention, the term "alkyl" refers to a saturated, straight or branched alkyl, which comprises preferably 1-10 carbon atoms, and more preferably 1-4 carbon atoms. Examples of alkyl include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, 2-ethylbutyl, n-pentyl, isopentyl, 1-methylpentyl, 1,3-dimethylbutyl, n-hexyl, 1-methylhexyl, n-heptyl, isoheptyl, 1,1,3,3-tetramethylbutyl, 1-methylheptyl, 3-methylheptyl, n-octyl, 2-ethylhexyl, 1,1,3-trimethylhexyl, 1,1,3,3-tetramethylpentyl, nonyl, decyl, 1,1,3,3,5,5-hexamethylhexyl, or the like.

[0048] In the present invention, the term "alkoxyl" or "alkoxy" as used herein means a group having a formula "-O-alkyl," wherein the definition of the "alkyl" in said formula has the meaning of "alkyl" as stated above.

[0049] In the present invention, the term "halogen" or "halo" denotes fluorine, chlorine, bromine or iodine.

[0050] In the present disclosure, the term "therapeutically acceptable salt" refers to salts or zwitterions of pharmaceutical compounds which are water or oil-soluble or dispersible, suitable for treatment of disorders and effective for their intended use. The salts may beprepared, for instance, during the final isolation and purification of the compounds or separately by reacting an amino group of the compounds with a suitable acid.

[0051] In the present disclosure, the term "isomer" refers to compounds having the same atomic composition (molecular formula) but different stereochemical formulae. Examples of isomers include, but are not limited to, stereoisomers (such as enantiomers and diastereomers), tautomers and optical isomers.

[0052] The term "prodrug" as used herein refers to a pharmacologically non-active substance that can be metabolized within the body into a pharmacologically active drug after intake. Examples of forms of a prodrug include, but are not limited to, a cleavable ester moiety, a cleavable carboxylic moiety, etc.

[0053] The term "solvate" as used herein refers to a solvent molecule(s) surrounding or complexing to the active compound. Examples of solvates include, but are not limited to, methanolate, ethanolate, trifluoroacetic acid (TFA) solvate, etc.

[0054] The term "hydrate" as used herein refers to a water molecule(s) surrounding or complexing to the active compound, e.g., a hydrated form or water molecule(s) present in the solid form of the active compound.

[0055] The term "treatment" or "treating" as used herein includes the alleviation, prevention, reversal, amelioration or control of a pathology, disease, disorder, process, condition or event, such as diabetes, or the symptoms of such pathology, disease, disorder, process, condition or event.

[0056] HDAC inhibitors are the most intensely pursued research area in the field of antitumor scaffold construction owing to their ability to induce the acetylation of histones and non-histone proteins involved in the regulation of gene expression and in various cellular pathways. Accordingly, HDAC inhibitors have also been explored to ascertain their potential in treating cancer. Particularly, HDAC6, a member of the HDAC family, has emerged as a promising target for cancer drug development due to its significant role in oncogenic celltransformation, particularly in its major substrate, α-tubulin. The HDAC6 gene, located in Xp11.23, encodes a protein with 1,215 amino acids. Overexpression of HDAC6 has been linked to tumorigenesis and improved survival. HDAC6 exerts its function through both deacetylation-dependent and -independent mechanisms. For instance, it plays a crucial role in the ubiquitin proteasome system by promoting the proteasomal degradation of HSP90. Additionally, it regulates various cellular processes such as cell migration, invasion, adhesion, and microtubule and skeletal dynamics through the deacetylation of tubulin and cortactin. Furthermore, HDAC6 is involved in the apoptosis pathway through the deacetylation of Ku70. HDAC6 also plays a role in controlling the expression of several proteins that are important in the human immune system, including PD-1, PD-L1, and tumor associated antigens. These proteins have become important targets for immune therapy in the treatment of cancer, and antibodies like nivolumab and pembrolizumab, which block these proteins, have shown encouraging results in fighting cancer and have been approved for use in tumor therapy.

[0057] For example, enriching the chemical toolbox of anti-GBM scaffolds requires the identification of new tractable targets. Explorations in this direction have led to several revelations advocating the candidature of cytochrome P45017A1 (CYP17A1) as an appealing drug target for GBM. Some seminal disclosures in this context are a) correlation of CYP17A1 overexpression with poor prognosis and decreased sensitivity to temozolomide in patients with GBM b) CYP17A1-mediated neurosteroid dehydroepiandrosterone (DHEA) production leading to temozolomide resistance in glioma cells c) upregulation of CYP17A1 leading to temozolomide (TMZ) resistance in glioma cells via promotion of DNA repair pathways and cell survival d) CYP17A1 mediated activation of the angiogenic factor (VEGF vascular epithelial growth factor) leading to tumor development and TMZ resistance e) interconnection between androgen receptors that acts as a transcription factor regulating the expression of genes involved in GBM cell growth and CYP17A1. In light of these revelations, steering thewheels of the anti-GBM drug discovery programs via rational design of CYP17A1 inhibitors appears to be a sagacious strategy.

[0058] The present disclosure assembles new anti-cancer chemical architectures furnished via structural enablement of the histone deacetylase (HDAC) inhibitory pharmacophore templates [(Surface recognition part (SRP) – linker - zinc binding group (ZBG)]. In particular, attempts have been made to target the HDAC6 isoform via small molecule HDAC6 inhibitors / abrogators as its apparent potential surpasses that of other HDAC isoforms as a target for cancers.

[0059] The present disclosure considers the fabrication of both single-target and dual-target HDAC inhibitory scaffolds to extract anti-cancer efficacy. While both the former and the latter proved to be potentially promising, the relatively favorable outcome of the endeavors centered on the dual targeting HDAC inhibitors tilts the inclination towards the approach of accomplishing multi-targeting anti-cancer templates. In addition, considering the emerging candidature of CYP17A1 as a druggable GBM target, the present disclosure conceives that installation of a CYP17A1 inhibitor within the HDAC inhibitory model as the SRP surface recognition part can pay dividends in the context of substantial anti-cancer efficacy. Accordingly, the present disclosure provides compounds for treating and / or preventing disease or disorder mediated by CYP17A1, HDAC (such as HDAC6) or both. Particularly, the compounds of the present disclosure are HDAC6-selective inhibitors.

[0060] In one embodiment, the compound of Formula (I) or (IA) may be selected from the group consisting of Compounds (12) to (15) and (24) to (27). In one embodiment, the compound of Formula (I) or (IB) may be selected from the group consisting of Compounds (16) to (23). In one embodiment, the compound of Formula (I) or (I') may be selected from the group consisting of Compounds (12) to (19). In one embodiment, the compound of Formula (I) or (I") may be selected from the group consisting of Compounds (20) to (27). In one embodiment, the compound of Formula (I), (I'), (IA) or (IA') may be selected from thegroup consisting of Compounds (12) to (15). In one embodiment, the compound of Formula (I), (I"), (IA) or (IA") may be selected from the group consisting of Compounds (24) to (27). In one embodiment, the compound of Formula (I), (I'), (IB) or (IB') may be selected from the group consisting of Compounds (16) to (19). In one embodiment, the compound of Formula (I), (I"), (IB) or (IB") may be selected from the group consisting of Compounds (20) to (23). The structures of compounds (12) to (27) are depicted above.

[0061] In one embodiment, the compound of Formula (I) as described herein can be prepared via the following steps:

[0062] (a) reacting a compound of Formula (II)(II),

[0063] wherein X is O, NRaor S wherein Rais H or C1-5alkyl, preferably O or NH; wherein each of RA1, RA2, RA3 and RA4 is independently halogen, hydroxy, C1-5alkyl or haloC1-5alkyl; q, r, s and t each independently represent an integer ranging from 0 to 3, provided that the sum of q, r, s and t ranges from 0 to 5;

[0064] with a compound of Formula (III)(III),

[0065] wherein n is an integer ranging from 2 to 10, preferably 4 to 9, more preferably 5 to 8; and R1 is C1-3alkoxy, preferably R1 is methoxy,

[0066] to give a compound of Formula (IV)

[0067] wherein X, n and R1 are as defined above, and

[0068] (b) converting the compound of Formula (IV) to the compound of Formula (I).

[0069] In one embodiment, step (b) of the method comprises reacting the compound of Formula (IV) with a compound of formula (V):

[0070] wherein R is H, C1-5alkyl, -C(=O)Rb, -C(=O)ORbor –C(=O)-NRbwherein Rbis H or C1-5alkyl, preferably R is H,

[0071] to give the compound of Formula (I).

[0072] In one embodiment, the compound of Formula (II) is selected from the group consisting of Formula (IIA), (IIB), (IIA'), (IIB'), (IIA") and (IIB"):

[0073] wherein X, RA1, RA2, RA3, RA4, q, r, s and t are as defined above.

[0074] In one embodiment, the compound of Formula (II) is Compound (8) or (9).

[0075] In one embodiment, the compound of Formula (II) is Compound (10) or (11).

[0076] In one embodiment, the process comprises a step of inverting a compound of Formula (IIA') to a compound of Formula (IIB'). In one embodiment, this inversion is achieved by a Mitsunobu reaction.

[0077] In one embodiment, the process comprises a step of converting a compound of Formula (IIA') to a compound of Formula (IIA"). In one embodiment, this conversion is achieved by sulfonylation, followed by azidization and reduction of the azide.

[0078] In one embodiment, the process comprises a step of converting a compound of Formula (IIB') to a compound of Formula (IIB"). In one embodiment, this conversion is achieved by sulfonylation, followed by azidization and reduction of the azide.

[0079] In one embodiment, the conversion of the compound of Formula (IV) to the compound of Formula (I) in step (b) is achieved by reacting the compound of Formula (IV) with hydroxylamine.

[0080] In one embodiment, the compound of Formula (IV) is selected from the group consisting of Formula (IVA), (IVB), (IVA'), (IVB'), (IVA") and (IVB"):Formula (IVA):Formula (IVB):Formula (IVA'):Formula (IVB'):Formula (IVA"):Formula (IVB"),

[0081] wherein X, n, R1, RA1, RA2, RA3, RA4, q, r, s and t are as defined above

[0082] In one embodiment, the process of preparing the compound of Formula (I) is based on Scheme 1, 2, 3 or 4:

[0083] Scheme 1:,

[0084] (a) alkoxyoxoalkanoic acids (n= 5-8), DCC, DIMAP, DCM, rt., 5h; (b) 50% Hydroxylamine (aq), DBU, MeOH, rt, 30 min.

[0085] Commercially available abiraterone (8) was subjected to a Stiglich esterification reaction for condensation with various alkoxy alkanoic acids employing the carbodiimide-mediated methodology. The resulting intermediates (28-31) were then treated with hydroxylamine in the presence of DBU to furnish the target compounds (12-15).

[0086] Scheme 2:,

[0087] (a) Benzoic acid, (C6H5)3P, DIAD, tetrahydrofuran (THF) rt, 3h; (b) sodium methoxide solution in MeOH, DCM: MeOH, rt, overnight; (c) alkoxy oxoalkanoic acids (n= 5-8), DCC, DIMAP, DCM, rt, 5h; (d) 50% Hydroxylamine (aq), DBU, MeOH, rt, 30 min.

[0088] For the inversion of the configuration at position 3 (3β-alcohol), starting material (8) was subjected to the Mitsunobu reaction using benzoic acid, diisopropyl azodicarboxylate (DIAD), and triphenylphosphine (TPP) in THF to generate the benzoate ester (32). The benzoate esters were then hydrolyzed with sodium methoxide in methanol to obtain the compound (9) (3α-alcohol). Subsequently, the compound (9) was esterified with alkoxy alkanoic acids of varied lengths to produce the intermediates (33-36), which were further treated with hydroxylamine in the presence of DBU to accomplish the synthesis of target compounds (16-19).

[0089] Scheme 3:,

[0090] (a) CH3SO2Cl, pyridine, stirring, rt. (b) NaN3, dimethylformamide (DMF), stirring, 80 °C (c) triphenylphosphine (PPh3), CH3OH, THF, stirring, 50 °C (d) various alkoxyalkanoic acids, EDC.HCl, HOBt, DIPEA, DMF, rt.; e) (i) LiOH (aq.), dioxane, rt; (ii) NH2OTHP, EDC.HCl, HOBt, DIPEA, DMF, rt; (iii) TFA (10% aq.), CH3OH, rt.

[0091] The route began with sulfonylation of abiraterone (8) with methane sulfonyl chloride in pyridine to yield compound (37). The intermediate (37) was subjected to azidation employing sodium azide in DMF. The azide intermediate (38) was further reduced to amine (10) using triphenylphosphine, and the amine obtained was amidated with alkoxy alkanoicacids using the EDC / HOBt-based methodology to yield compounds (39-42). The esters (39-42) were hydrolyzed using LiOH to afford carboxylic acids, which were subsequently treated with NH2OTHP to form the amides, and the tetrahydropyranyl functionality of the amides was cleaved using TFA, resulting in the formation of corresponding hydroxamic acids (20-23).,

[0093] (a) CH3SO2Cl, pyridine, stirring, rt; (b) NaN3, DMF, stirring, 80 °C; (c) PPh3, CH3OH, THF, stirring, 50 °C; (d) various alkoxy alkanoic acids, EDC.HCl, HOBt, DIPEA, DMF, rt; e) (i) LiOH (aq.), dioxane, rt; (ii) NH2OTHP, EDC.HCl, HOBt, DIPEA, DMF, rt; (iii) TFA (10% aq), CH3OH, rt.

[0094] The same reaction sequence as described in scheme 3 was employed with the only point of difference being the starting material. 3α- alcohol (9) was used as the starting precursor in this scheme and the desired adducts were attained in satisfactory yields (24-27).

[0095] The applications or uses of the compounds or derivatives thereof or composition / combination thereof described herein relate to the treatment of diseases / disorders cancers, including solid cancers and hematological neoplasms. In one embodiment, thecancer is a metastatic cancer. Examples of the cancer include, but are not limited to, lymphoma, myeloma, leukemia, lung cancer (such as non-small cell lung carcinoma (NSCLC), breast cancer, cervical cancer, colorectal cancer (CRC), pancreatic cancer, prostate cancer, stomach cancer, renal cancer and brain cancer. In some embodiments, the brain cancer is a primary brain tumor or a metastatic brain cancer, for example glioma or meningioma, or more specifically glioblastoma, medulloblastoma, or acoustic neuroma.

[0096] Histone deacetylase (HDAC) inhibitors are a class of anti-cancer agents that play important roles in epigenetic or non-epigenetic regulation, inducing death, apoptosis, and cell cycle arrest in cancer cells.

[0097] Cytochrome P450 17A1 (steroid 17α-monooxygenase, 17α-hydroxylase, 17-alpha-hydroxylase, 17,20-lyase, 17,20-desmolase) is an enzyme of the hydroxylase type that in humans is encoded by the CYP17A1 gene on chromosome 10. HDACs have been found to promote GBM development by the action of phosphorylated sp1 induced by deacetylation (HDAC 1 / 2). In particular, sp1 has recently been identified as a novel substrate for HDAC6, and GBM is reported to hijack sp1 for its transcriptional activity through HDAC1 / 2 / 6's deacetylation, promoting the self-renewal of malignancy based on the upregulation of B cell-specific Mo-MLV integration site 1 (BMI1) and human telomerase reverse transcriptase (hTERT), along with regulation of G2 / M progression and DNA repair through altered gene transcription.

[0098] Without being bound to theory, the compounds and derivatives thereof disclosed herein may be suitable for treating various types of cancer, including those where HDAC6 and CYP17A1 play critical roles in promoting cancer cell proliferation and survival. For example, HDAC6 is implicated in several cancers, including breast cancer, colorectal cancer, lung cancer, and hematological malignancies. In these cancers, HDAC6 contributes to cancer progression and drug resistance, making it an attractive target for therapeutic intervention. Dual inhibition of HDAC6 and CYP17A1 by the compounds and derivativesdisclosed herein could offer a unique advantage in these cancer types by simultaneously targeting key pathways involved in tumor growth and treatment resistance.

[0099] While it may be possible for the compounds of the present disclosure to be administered as the raw chemical, it is also possible to present them as a pharmaceutical composition or formulation. Accordingly, the present disclosure provides a pharmaceutical formulation or composition comprising a compound or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, together with one or more pharmaceutically acceptable carriers thereof and optionally one or more other therapeutic ingredients. The carrier(s) must be "acceptable" in the sense of being compatible with the other ingredients of the composition or formulation and not deleterious to the recipient thereof. Proper composition / formulation is dependent upon the route of administration chosen. Compositions / formulations may take the form of tablets, pills, capsules, semisolids, powders, sustained release formulations, solutions, suspensions, elixirs, aerosols, or any other appropriate compositions; and comprise at least one compound of this disclosure in combination with at least one pharmaceutically acceptable excipient. Suitable excipients are well known to persons of ordinary skill in the art, and they, and the methods of formulating the compositions / formulations, may be found in such standard references as Remington: The Science and Practice of Pharmacy, A. Gennaro, ed., 20th edition, Lippincott, Williams & Wilkins, Philadelphia, Pa. Suitable liquid carriers, especially for injectable solutions, include water, aqueous saline solution, aqueous dextrose solution, and glycols. The pharmaceutical compositions / formulations of the present disclosure may be manufactured in a manner that is itself known, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or compression processes.

[0100] The compositions / formulations include those suitable for oral, parenteral (including subcutaneous, intradermal, intramuscular, intravenous, intraarticular, and intramedullary), intraperitoneal, transmucosal, transdermal, rectal and topical (including dermal, buccal, sublingualand intraocular) administration, although the most suitable route may depend, for example, upon the condition and disorder of the recipient. Oral administration is a preferred route. The compositions / formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. All methods include the step of bringing into association a compound of the present disclosure or a pharmaceutically acceptable salt, prodrug or solvate thereof ("active ingredient") with the carrier which constitutes one or more accessory ingredients. In general, the compositions / formulations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both and then, if necessary, shaping the product into the desired composition / formulation.

[0101] For oral administration, suitable pharmaceutical compositions / formulations of the present disclosure include powders, granules, pills, tablets, lozenges, chews, gels, and capsules as well as liquids, syrups, suspensions, elixirs, and emulsions. These compositions / formulations may also include anti-oxidants, flavorants, preservatives, and suspending, thickening and emulsifying agents, colorants, flavoring agents and other pharmaceutically acceptable additives. Formulations for oral administration may be formulated to be immediate release or modified release, where modified release includes delayed, sustained, pulsed, controlled, targeted and programmed release.

[0102] For parenteral administration, the compounds or compositions / formulations of the present disclosure are administered directly into the blood stream, into muscle, or into an internal organ via an intravenous, intraarterial, intraperitoneal, intramuscular, subcutaneous or other injection or infusion. Parenteral formulations may be prepared in aqueous injection solutions which may contain, in addition to the compound of the disclosure, buffers, antioxidants, bacteriostats, salts, carbohydrates, and other additives commonly employed in such solutions. Parenteral administrations may be immediate release or modified release (such as an injected or implanted depot).

[0103] Compounds or compositions / formulations of the present disclosure may also be administered topically, (intra)dermally, or transdermally to the skin or mucosa. Typical formulations include gels, hydrogels, lotions, solutions, creams, ointments, dressings, foams, skin patches, wafers, implants and microemulsions. Compounds or compositions / formulations of the present disclosure may also be administered via inhalation or intranasal administration, such as with a dry powder, an aerosol spray or as drops. Additional routes of administration for compounds of the present disclosure include intravaginal and rectal (by means of a suppository, pessary or enema), and ocular and aural.

[0104] The following examples are provided to make the present disclosure more comprehensible to those of ordinary skill in the art to which the present disclosure pertains, but are not intended to limit the scope of the disclosure. EXAMPLES

[0105] Abbreviations GBM Glioblastoma TMZ Temozolomide CYP17A1 Cytochrome P45017A1 DHEA Dehydroepiandrosterone HDAC Histone deacetylase UPLC-MS Ultra performance liquid chromatography-mass spectrometry DHR Dihydrorhodamine 123 SRP Surface recognition part CDH23 Cadherin Related 23 LRMP Lymphoid-restricted membrane protein LCP1 Lymphocyte Cytosolic Protein 1 IFI44L Interferon Induced Protein 44 LikeSLC25A27 Solute Carrier Family 25 Member 27 NPY1R Neuropeptide Y Receptor Y1 SLITRK1 SLIT and NTRK Like Family Member 1 SOD Sodium dismutase GPX Glutathione peroxidase

[0106] Experiments and Materials

[0107] Bruker DRX-300 and 600 spectrometers were used for obtaining the1H and13C NMR spectra. High-resolution mass spectra (HRMS) were measured with a JEOL (JMS-700) electron impact (EI) mass spectrometer. Shimadzu LC-2030C with UV / vis detector SPD-20 A using C-18 column (Agilent ZORBAX Eclipse XDB-C 18, 5 μm- 4.6 × 150 mm) was used for determining the final purity of the compounds, and all compounds are >95% pure by HPLC analysis. All reactions were carried out under an atmosphere of dry nitrogen.

[0108] Cell Culture and treatment

[0109] GBM cell lines, A172, LN229, T98G, U373MG and U87MG, were purchased from American Type Culture Collection (Manassas, VA, USA). Patient-derived GBM cell, Pt#3, was established based on previous studies (see, e.g., BMC cancer 2021, 21, 1-9). Cells were cultured in DMEM supplemented with 10% fetal bovine serum and 100 mg / ml streptomycin and penicillin. The procedure for generating TMZ-resistant cells, including A172-R, Pt#3-R and U87MG-R, was also established based on previous studies (see, e.g., Oncogenesis 2017, 6 (5), e339-e339). These resistant cells were culture in the presence of 50 (A172-R and Pt#3-R) or 100 mM (U87MG-R) TMZ (MilliporeSigma Corporate, St. Louis, MO, USA). 22rv1 cell lines were used as a prostate cancer xenograft model.

[0110] HDAC6 knockout and CYP17A1 knockdown

[0111] CRISPR-mediated HDAC6 knockout in U87MG cells were serviced by BIOTOOLS Co., Ltd. (New Taipei City, Taiwan). CYP17A1 siRNA was purchased fromDharmacon Inc. (Lafayette, CO, USA). After transfection with siRNA using DharmaFECT™ transfection reagent for 72 h, cells were treated with the indicated drug for IC50 analysis.

[0112] Patient specimens and immunohistochemistry (IHC)

[0113] Characteristics of patient specimens were clearly provided in the previous study. IHC was performed using the VECTASTAIN® ABC Kit (Newark, CA, USA).

[0114] Steroidomic metabolomics using UPLC-MS

[0115] The detailed procedure was described in previous studies (see, e.g., Free Radical Biology and Medicine 2021, 172, 430-440).

[0116] CCK8 cell viability assay

[0117] The CCK8 reagent was purchased from TargetMOI (Wellesley Hills, MA, USA) and used according to the manufacturer’s instruction.

[0118] CYP17A1 activity assay

[0119] CYP17A1 is a rate-limiting enzyme responsible for DHEA production. Hence, we measured DHEA levels to represent CYP17A1 activity. DHEA levels were estimated using DHEA ELISA kit (ADI-900-093; Enzo Life Sciences, Inc., Farmingdale, NY, USA) according to the manufacturer’s instruction.

[0120] Molecular Docking through iGEMDOCK Software

[0121] Structure-based molecular docking was performed to screen compound 12 based on the binding energy. The docking process was carried out between protein and compounds with a population size of 800, generations of 70, and solutions of 10 through iGEMDOCK version 2.1. Protein structures of CYP17A1 and HDAC6 were obtained from Protein Data Bank (3RUK and 5EDU).

[0122] Western blotting analysis

[0123] Briefly, after electrophoresis and protein transfer, the polyvinylidene difluoride membrane (MilliporeSigma Corporate) was incubated with the first antibody, anti-GFP (GeneTex International Corporation, HsinChu, Taiwan), anti-DDK (GeneScript, Piscataway,NJ, USA), anti-HDAC6 (Santa Cruz Biotechnology, Inc., Dallas, TX, USA), anti-CYP17A1 (Proteintech Group, Inc., Rosemont, IL, USA), anti-Rad51 (Abcam, Cambridge, UK), anti-γH2Ax (Abcam), SOD1 (GeneTex), SOD2 (GeneTex), GPX1 (GeneTex), GPX4 (GeneTex), ^-tubulin (GeneTex) or anti-GAPDH (GeneTex) antibody overnight at 4℃. After incubating with the HRP-conjugated secondary antibody for 1 h at room temperature, the membrane was incubated with enhanced chemiluminescence (MilliporeSigma Corporate), and subjected to an image analyzer.

[0124] Cell cycle analysis

[0125] After fixation in the 70% ethanol at -20℃ overnight, cells were stained with propidium iodide (PI) in the presence of RNase for 30 min at room temperature. Stained cells were analyzed using flow cytometry and the Guava software (MilliporeSigma Corporate).

[0126] RNA-seq

[0127] RNA was isolated using the RNA extracting kit (Zymo Research, Irvine, CA, USA), and subjected to RNA-seq serviced by BIOTOOLS Co., Ltd.

[0128] Immunofluorescence for γH2Ax

[0129] Cells were seeded onto poly-l-lysine-coated coverslips. After treatment, cells were fixed and permeabilized using 4% paraformaldehyde and 0.5% triton x-100, respectively. Subsequently, blocked cells on the coverslip were incubated with the anti-γH2Ax antibody followed by the incubation with FITC-conjugated secondary antibody. Cell foci labeled by γH2Ax were estimated using a fluorescent microscope.

[0130] ROS assay and MitoSOX assay

[0131] Dihydrorhodamine 123 (DHR; Thermo Fisher Scientific, Waltham, MA, USA) was used to label cellular ROS detected by the flow cytometry, and the MitoSOX reagent (Thermo Fisher Scientific) was used to label mitochondrial ROS.

[0132] Animal experiment

[0133] Xenograft model

[0134] Animal experiments were approved by the institutional animal care and use committee of Taipei Medical University (LAC-2020-0303).

[0135] NOD.CB17-Prkdcscid / NCrCrl mice (8-week-old) were purchased from BioLASCO Taiwan Co., Ltd. (Taipei, Taiwan). Cells (1x106) in 50 µl DMEM were injected onto the back of mice. After 10 days, the tumor was touchable. Mice were injected with DMSO (the control group), abiraterone (8) or compound 12 twice per week.

[0136] Othortopic model

[0137] Cells (5x105) in 5 ml DMEM were injected into the brain of mice. After 10 days, the tumor was touchable. Mice were injected with DMSO (the control group) or compound 12 twice per week. For oral administration, a suspension containing compound 12 was prepared with a solution containing 40% of ethyl oleate, 12% of lecithin, 28% of tween 80, and 20% of propylene glycol. Different doses of compound 12 were prepared in 100 µl solution for oral administration.

[0138] Statistical analysis

[0139] Experiments were performed 3 times independently, and data were expressed as mean±s.e.m. P value < 0.05 was considered as the significant difference.

[0140] Synthetic Examples

[0141] Examples 1-4: Synthesis of Compounds (12) to (15)

[0142] Synthesis of 1-((3S,10R,13S)-10,13-dimethyl-17-(pyridin-3-yl)- 2,3,4,7,8,9,10,11,12,13,14,15-dodecahydro-1H-cyclopenta[a]phenanthren-3-yl) 7-methyl heptanedioate (28)

[0143] A mixture of compound 8 (100 mg, 0.00029 mol), DCC (109mg, 0.00057 mol), DIMAP (17 mg, 0.00014 mol) in DCM (2 mL), and 7-methoxy-7-oxo heptanoic acid (80 mg, 0.00043 mol) was stirred at room temperature for 5h. The reaction mixture was quenched with water and extracted with EtOAc (50 mL x 3). The combined organic layer was driedover anhydrousMgSO4,concentrated under reduced pressure, and purified by silica gelchromatography (EtOAc : Hexane: 2:2) to give 28 in 71 % yield;1H NMR (300 MHz, CD3OD):δ 1.07 (s, 3 H), 1.10 (s, 3 H), 1.11-1.20 (m, 2 H), 1.31-1.37 ( m, 2 H), 1.46-1.52 (m, 1H), 1.58-1.63 (m, 6H), 1.65-1.73 (m, 3H), 1.76-1.85( m, 2H), 1.89 -1.93 (m, 1H), 2.03-2.12 (m, 5H), 2.26-2.33 (m, 5H), 3.64 (s, 3H), 4.50-4.57 (m, 1H), 5.42-5.43 (m, 1H), 6.07-6.08 (m, 1H), 7.36 (dd, J = 4.9 & 7.9 Hz, 1H), 7.81-7.83 (m, 1H), 8.36 (dd, J = 1.1 & 4.8 Hz, 1H), 8.51 (d, J = 1.6 Hz, 1H).

[0144] Synthesis of 1-((3S,10R,13S)-10,13-dimethyl-17-(pyridin-3-yl)- 2,3,4,7,8,9,10,11,12,13,14,15-dodecahydro-1H-cyclopenta[a]phenanthren-3-yl) 8-methyl octanedioate (29)

[0145] The title compound 29 was synthesized using compound 8 and 8-methoxy-8-oxooctanoic acid in a manner similar to that described forcompound 28 in 70 %yield;1H NMR (300 MHz, CD3OD): δ 1.09 (s, 3 H), 1.12 (s, 3 H), 1.34-1.38 (m, 8 H), 1.60 – 1.65 (m, 9 H), 1.84-1.92 (m, 3 H), 2.07-2.15 (m, 3 H), 2.31 – 2.33 (m, 6 H), 3.67 (s, 3 H), 4.55 (m, 1 H), 5.44 (d, J = 4.8 Hz, 1 H) 6.11 (m, 1 H), 7.39 (m, 1 H), 7.85 (m, 1 H), 8.42 (bs, 1 H), 8.57 (bs, 1 H)

[0146] Synthesis of 1-((3S,10R,13S)-10,13-dimethyl-17-(pyridin-3-yl)- 2,3,4,7,8,9,10,11,12,13,14,15-dodecahydro-1H-cyclopenta[a]phenanthren-3-yl) 9-methyl nonanedioate (30).

[0147] The title compound 30 was synthesized using compound 8 and 9- methoxy-9-oxononanoic acid in a manner similar to that described forcompound 28 in 72 %yield;1H NMR (300 MHz, CD3OD): δ 1.01 (s, 3 H), 1.05 (s, 3 H), 1.27 (bs, 9 H), 1.56 -1.62 (m, 10 H), 1.66-1.85 (m, 4 H), 1.98 -2.07 (m, 3 H), 2.21 -2.29 (m, 5 H), 3.62 (s, 3 H), 4.58 (m, 1 H), 5.38 (d, J = 4.8 Hz, 1 H), 5.96 (m, 1 H), 7.18 (m, 1 H), 7.617.8 Hz, 1 H), 8.41 (bs, 1 H), 8.59 (bs, 1 H).

[0148] Synthesis of 1-((3S,10R,13S)-10,13-dimethyl-17-(pyridin-3-yl)- 2,3,4,7,8,9,10,11,12,13,14,15-dodecahydro-1H-cyclopenta[a]phenanthren-3-yl) 10- methyl decanedioate (31)

[0149] The title compound 31 was synthesized using compound 8 and 10- methoxy-10-oxononanoic in a manner similar to that described for compound 28 in 70 % yield;1H NMR (300 MHz, CD3OD): 1.08 (s, 3 H), 1.10 (s, 3 H), 1.31(s, 7 H), 1.12-1.19 (m, 2 H), 1.49 (m, 1 H), 1.58-164 (m, 7 H), 1.66 -1.77 (m, 3 H), 1.77 -1.86 (m, 2 H), 1.91 (m, 1 H), 2.01-2.13 (m, 5 H), 2.23-2.34 (m, 5 H), 3.63 (s, 3 H), 4.53 (m, 1 H), 5.43 (d, J = 5.1 Hz, 1 H), 6.07 (m, 1 H), 7.35 (dd, J = 4.9 Hz & 7.9 Hz, 1 H), 7.82 (m, 1H), 8.36 (dd, J = 1 Hz & 4.7 Hz, 1H), 8.51 (d, J = 1.5 Hz, 1H).

[0150] Synthesis of (3S,10R,13S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15- dodecahydro-1H-cyclopenta[a]phenanthren-3-yl 7-(hydroxyamino)-7-oxoheptanoate (12)

[0151] In a suspension of the compound (28) (100 mg, 0.0002 mol) in methanol (0.5 ml), DBU (90 mg,0.00059 mol) and hydroxyl amine (0.06 ml, 0.002mol) were added and the mixture was stirred at room temperature for 30 min. The reaction mixture was quenched with water and extracted with EtOAc (50 mL x 3). The combined organic layer was dried over anhydrous MgSO4, concentrated under reduced pressure and was purified by silica gel chromatography (DCM: Methanol: 95:5) to give compound (12) in 10% yield; mp: 180-181 °C; HPLC purity: - 96.77%.1H NMR (300 MHz, CD3OD): δ 1.12 (s, 3 H), 1.15 (s, 3 H), 1.16-1.23 (m, 2 H), 1.32-1.44 (m, 4 H), 1.60-1.79 (m, 10 H), 1.84-1.97 (m, 3 H), 2.07-2.19 (m, 3 H), 2.33 – 2.37 (m, 5 H), 4.57 (m, 1 H), 5.47 (d, J = 4.0 Hz, 1 H), 6.12 (m, 1 H), 7.40 (m, 1 H) 7.87 (m, 1 H), 8.36 (dd, J = 1.1 Hz & 4.8 Hz, 1H), 8.51 (d, J = 1.7 Hz, 1H). 13CNMR (75 MHz, CDCl3): 16.58, 19.27, 20.82, 24.85, 25.32, 27.77, 28.80, 29.70, 30.40, 31.51, 31.82, 34.60, 35.20, 36.79, 36.92, 38.17, 47.33, 50.24, 57.47, 73.73, 122.27, 123.25, 129.52,133.21, 134.09, 140.05, 147.45, 151.47, 173.43. HRMS (ESI) for C31H43N2O4 (M + H+): calcd, 507.3223; found, 507.3225.

[0152] Synthesis of (3S,10R,13S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15- dodecahydro-1H-cyclopenta[a]phenanthren-3-yl 8-(hydroxyamino)-8-oxooctanoate (13)

[0153] The title compound (13) was synthesized using compound (29) in a manner similar to that described for compound (12) in 10 % yield; mp: 138-139 °C; HPLC purity: - 95.55 %.1H NMR (300 MHz, CD3OD): d 1.07 (s, 3 H), 1.11 (s, 3 H), 1.18 (m, 1 H), 1.36 (bs, 5 H), 1.52-1.66 (m, 11 H), 1.87-1.91 (m, 2 H), 2.05 - 2.18 (m, 5 H), 2.26-2.37 (m, 5 H), 4.64 (m, 1 H), 5.44 (d, J = 4.9 Hz, 1 H), 6.04 (bs, 1 H), 7.29 (m, 1H), 7.70 (d, J = 7.9 Hz, 1H), 8.49(s, 1H), 8.64 (s, 1H).13CNMR (75 MHz, CDCl3): 16.58, 19.27, 20.82, 22.63, 24.73, 27.22, 27.78, 28.58, 29.33, 29.71, 31.52, 31.82, 34.49, 35.21, 36.80, 36.92, 38.16, 47.34, 50.25, 57.47, 73.79, 122.31, 123.23, 129.51, 134.07, 140.03, 147.53, 151.50, 173.39. HRMS (ESI) for C32H45N2O4 (M + H+): calcd, 521.3379; found, 521.3374.

[0154] Synthesis of (3S,10R,13S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15- dodecahydro-1H-cyclopenta[a]phenanthren-3-yl 9-(hydroxyamino)-9-oxononanoate (14)

[0155] The title compound (14) was synthesized using compound (30) in a manner similar to that described for compound (12) in 10 % yield; mp: 127-128 °C; HPLC purity: - 95.63%.1H NMR (300 MHz, CD3OD): d 1.07 (s, 3 H), 1.11 (s, 3 H), 1.18 (m, 1 H), 1.33 (bs, 6 H), 1.50 -1.78 (m, 12 H), 1.87 - 1.91 (m, 2 H), 2.02 - 2.18 (m, 5 H), 2.27 - 2.37 (m, 5 H), 2.30-2.33 (m, 2 H), 4.63 (m, 1 H), 5.44 (d, J = 4.5 Hz, 1 H), 6.03 (bs, 1 H), 7.27 (s, 1 H), 7.70 (d, J= 7.8 Hz, 1 H), 8.48 (s, 1 H), 8.64 (s, 1 H).13C NMR (75 MHz, CDCl3): 16.56, 17.82, 19.25, 20.80, 21.29, 21.70, 23.11, 24.50, 25.11, 28.47, 30.37, 31.49, 31.80, 34.35, 35.18, 36.76, 38.12, 38.85, 47.30, 50.22, 57.44, 73.79, 122.25, 123.28, 129.56, 133.20, 134.13,140.00, 147.33, 151.37, 171.14, 173.26. HRMS (ESI) for C33H47N2O4 (M + H+): calcd, 535.3536; found, 535.3536.

[0156] Synthesis of (3S,10R,13S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15- dodecahydro-1H-cyclopenta[a]phenanthren-3-yl 10-(hydroxyamino)-10-oxodecanoate (15).

[0157] The title compound (15) was synthesized using compound (31) in a manner similar to that described for compound (12) in 10 % yield; mp: 79-80 °C; HPLC purity: - 95.98%.1H NMR (300 MHz, CD3OD): 1.08 (s, 3 H), 1.10 (s, 3 H), 1.12-1.90 (m, 2 H), 1.31 (s, 7 H), 1.49 (m, 1 H), 1.58-1.64 (m, 7 H), 1.66-1.73 (m, 3 H), 1.76-1.85 (m, 2 H), 1.91 (m, 1 H), 2.00-2.14 (m, 5 H), 2.26-2.29 (m, 3 H), 2.31-2.33 (m, 2 H), 4.53 (m, 1 H), 5.43 (d, J = 5 Hz, 1 H), 6.07 (m, 1 H), 7.35 (dd, J = 4.8 Hz & 7.9 Hz, 1 H), 7.82 (m, 1 H), 8.36 (dd, J = 1 Hz & 4.8 Hz, 1 H), 8.51 (d, J = 1.8 Hz, 1 H).13C (75 MHz, DMSO): 16.27, 18.53, 20.15, 24.68, 25.11, 25.61, 28.31, 28.50, 29.88, 31.03, 31.30, 32.60, 33.15, 33.98, 34.61, 35.80, 36.77, 40.10, 46.68, 48.61, 49.84, 57.10, 69.75, 121.16, 123.39, 129.04, 132.17, 133.34, 138.86, 147.21, 147.82, 151.03, 169.08, 172.36. HRMS (ESI) for C34H49N2O4(M + H+): calcd, 549.3692; found, 549.3695.

[0158] Examples 5-8: Synthesis of Compounds (16) to (19)

[0159] Synthesis of (3S,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15-dode cahydro-1H-cyclopenta[a]phenanthren-3-yl benzoate (32)

[0160] A mixture of compound (8) (5g, 0.0143 mol), benzoic acid (1.75g, 0.0143 mol), triphenylphosphine (3.93g, 0.015 mol), and diisopropylazodicarboxylate (3ml, 0.0149 mol) in THF was stirred at room temperature for 3 h. The reaction mixture was quenched with water and extracted with EtOAc (50 ml x 3). The combined organic layers were dried over anhydrous MgSO4 and concentrated under reduced pressure. The residue was purified by silica gel chromatography (n-Hexane: EtOAc = 92:8) to give a solid product (32) in 52.7%yield;1H NMR (300 MHz, CD3OD): δ 1.13, (s, 3H), 1.20 (s, 3H), 1.28 -1.60 (m, 5H), 1.71 -1.99 (m, 7H), 2.09-2.43 (m, 5H), 4.60 (bs, 1H), 5.39 (s, 1H), 6.12 (s, 1H), 7.39-7.43 (m, 3H), 7.56 (m, 1H), 7.88 (m, 1H), 7.98-8.06 (m, 2H), 8.414.8 Hz and 1.5 Hz, 1H), 8.56 (m, 1H).

[0161] Synthesis of (3R,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15-dode cahydro-1H-cyclopenta[a]phenanthren-3-ol (9).

[0162] Compound (32) (6g, 0.013 mol) was dissolved in DCM and methanol (50:50), and sodium methoxide 5.4 M (3ml, 0.055 mol) was added to the solution. The reaction mixture was stirred overnight at room temperature and then quenched with water and extracted with EtOAc (50 ml x 3). The combined organic layers were dried over anhydrous MgSO4 and concentrated under reduced pressure. The residue was purified by silica gel chromatography (n-Hexane: EtOAc = 8:2) to give a solid product (9) in 57.6% yield;1H NMR (300 MHz, CDCl3): δ 1.08 (s, 3H), 1.10 (s, 3H), 1.28 -1.53(m, 5H), 1.67-1.83 (m, 7H), 2.63 (m, 5H), 4.06 (bs, 1H), 5.48 (bs, 1H), 6.03 (dd, J = 3.3 and 1.8 Hz, 1H), 7.25 (m, 1H), 7.68 (m, 1H), 8.49 (dd, J = 4.8 and 1.5 Hz, 1H), 8.65 (d, J = 1.8 Hz, 1H).

[0163] Synthesis of 1-((3R,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15-d odecahydro-1H-cyclopenta[a]phenanthren-3-yl) 7-ethyl heptanedioate (33)

[0164] A mixture of compound (9) (240 mg, 0.0006 mol), 7-ethoxy-7-oxoheptanoic acid (193 mg, 0.0010 mol), DCC (354 mg, 0.0017 mol), and DIMAP (41 mg, 0.0003 mol) in DCM (10 ml) was stirred at room temperature for 5 hours and then quenched with water and extracted with EtOAc (50 ml x 3). The combined organic layers were dried over anhydrous MgSO4and concentrated under reduced pressure. The residue was purified by silica gel chromatography (n-hexane: EtOAc = 7.8: 2.2) to give a solid product (33) in 72.60% yield;1H NMR (300 MHz, CDCl3): δ 1.13 (s, 6H), 1.24-1.42 (m, 11H), 1.64-1.81 (m, 12H),2.31-2.33 (m,7H), 4.13 (t, J = 7.2 Hz, 2H), 5.03 (bs, 1H), 5.39 (d, J = 5.4 Hz, 1H), 6.24 (s, 1H), 7.61 (bs, 1H), 8.09 (d, J = 7.8 Hz, 1H), 8.57 (s, 1H), 8.71 (s, 1H).

[0165] Synthesis of 1-((3R,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15-d odecahydro-1H-cyclopenta[a]phenanthren-3-yl) 8-methyl octanedioate (34).

[0166] The compound (34) was obtained in 64% yield using compound (9), and 8-methoxy-8-oxooctanoic acid in a manner similar to that described for the synthesis of compound (33);1H NMR (300 MHz, CDCl3): δ 1.10 (s, 6H), 1.28(s, 3H), 1.32-1.39 (m, 10H), 1.62-1.69 (m, 6H), 1.70 - 2.10(m, 4H), 2.20 - 2.34 (m, 6H), 3.68 (s, 3H), 5.03 (bs, 1H), 5.32(d, J = 4.8 Hz, 1H), 6.25 (bs, 1H), 7.64 (m, 1H), 8.11 (d, J = 7.8 Hz, 1H), 8.57 (d, J = 4.2 Hz, 1H), 8.72(s, 1H).

[0167] Synthesis of 1-((3R,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15-d odecahydro-1H-cyclopenta[a]phenanthren-3-yl) 9-methyl nonanedioate (35).

[0168] The compound (35) was obtained in 71% yield using compound (9) and 9- methoxy-9-oxo nonanoic acid in a manner similar to that described for the synthesis of compound (33);1H NMR (300 MHz, CDCl3): δ 1.09 (s, 3H), 1.10 (s, 3H), 1.31-1.33 (m, 12H), 1.61-1.65 (m, 6H), 1.76-1.81 (m, 6H), 2.06-2.13 (m, 2H), 2.29-2.35 (m, 5H), 3.68 (s, 3H), 5.03 (t, J = 2.8 Hz, 1H), 5.33 (s, 1H), 6.14 (bs, 1H), 7.44 (dd, J = 7.2 and 5.1 Hz, 1H), 7.90 (d, J = 8.1 Hz, 1H), 8.52 (d, J = 4.5 Hz, 1H), 8.68 (s, 1H).

[0169] Synthesis of 1-((3R,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15-d odecahydro-1H-cyclopenta[a]phenanthren-3-yl) 10-methyl decanedioate (36).

[0170] The compound (36) was obtained in 74% yield using compound (9), and 10- methoxy-10-oxo nonanoic acid in a manner similar to that described for the synthesis of compound (33);1H NMR (300 MHz, CDCl3): δ 1.09 (s, 3H), 1.11 (s, 3H), 1.21-1.31 (m, 12H),1.60-1.81 (m, 14H), 2.05-2.35 (m, 7H), 3.68 (s, 3H), 5.03 (t, J = 2.7 Hz, 1H), 5.32 (bs, 1H), 6.13 (dd, J = 3.0 and1.8 Hz, 1H), 7.41 (m, 1H), 7.87 (m, 1H), 8.51 (dd, J = 5.1 and 1.4 Hz, 1H), 8.67 (d, J = 1.8 Hz, 1H).

[0171] Synthesis of (3R,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15-dode cahydro-1H-cyclopenta[a]phenanthren-3-yl 7-(hydroxyamino)-7-oxoheptanoate (16).

[0172] A mixture of compound (33) (310 mg, 0.0005 mol), hydroxylamine (aq) (0.2 ml, 0.005 mol), and DBU (0.27 ml, 0.0017 mol) in methanol was stirred at room temperature for 30 min. The reaction mixture was quenched with water and extracted with EtOAc (50 ml x 3). The combined organic layers were dried over anhydrous MgSO4 and concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM: CH3OH = 95:5) to give a solid product (16) in 15.8% yield; mp: 148-149 °C HPLC purity: - 97.57%.1H NMR (300 MHz, DMSO): δ 1.03 (s, 3H), 1.05 (s, 3H), 1.24 (bs, 5H), 1.44-1.93 (m, 14H), 2.00-2.22 (m, 8H), 4.91 (s, 1H), 5.26 (d, J = 4.5 Hz, 1H), 6.13 (s, 1H), 7.33-7.37 (m, 1H), 7.77 (m, 1H), 8.44 (dd, J = 4.8 and 1.5 Hz, 1H), 8.60 (d, J = 1.8 Hz, 1H), 8.68 (s, 1H), 10.35 (s, 1H).13C (75 MHz, CD3OD): 15.54, 17.86, 20.29, 24.46, 24.99, 25.73, 28.14, 30.28, 31.22, 31.29, 32.15, 33.35, 33.92, 35.04, 35.89, 36.96, 47.10, 50.53, 57.67, 70.71, 121.50, 123.48, 129.55, 133.59, 134.59,138.82, 146.55, 146.68, 151.37, 171.34, 173.54. HRMS (ESI) for C31H43N2O4 (M + H+): calcd, 507.3233; found, 507.3225.

[0173] Synthesis of 1-((3R,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15-d odecahydro-1H-cyclopenta[a]phenanthren-3-yl) 8-methyl octanedioate (17).

[0174] The compound (17) was obtained in 16.8% yield using compound (34) in a manner similar to that described for the synthesis of compound (16); mp: 105-106 °C HPLC purity: - 95.12%.1H NMR (300 MHz, DMSO): δ 1.03 (s, 3H), 1.06 (s, 3H), 1.24 (bs, 11H), 1.47-1.49 (m, 7H), 1.65-1.71 (m, 6H), 1.93 – 2.20 (m, 6H), 4.91(bs, 1H), 5.26 (d, J = 5.1 Hz,1H), 6.14(s, 1H), 7.37 (dd, J = 7.8 and 4.5 Hz, 1H), 7.78 (m, 1H), 8.45 (m, 1H), 8.61 (s, 1H), 10.33 (s, 1H).13C (75 MHz, CD3OD): 15.57, 17.89, 20.30, 24.71, 25.17, 25.75, 28.36, 28.43, 30.28, 31.23, 31.32, 32.30, 33.37, 34.05, 35.03, 35.90, 36.96, 47.11, 50.53, 57.68, 70.68, 121.48, 123.63, 129.76, 133.72, 134.90, 138.83, 146.27, 146.42, 151.24, 171.42, 173.64. HRMS (ESI) for C32H45N2O4 (M + H+): calcd, 521.3379; found, 521.3380.

[0175] Synthesis of (3R,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15-dode cahydro-1H-cyclopenta[a]phenanthren-3-yl 9-(hydroxyamino)-9-oxononanoate (18).

[0176] The compound (18) was obtained in 13.5% yield using compound (35) in a manner similar to that described for the synthesis of compound (16); mp: 122-123 °C HPLC purity: - 97.58%.1H NMR (300 MHz, DMSO): δ 1.03 (s, 3H), 1.05 (s, 3H), 1.24 (bs, 10H), 1.40-1.75 (m, 13H), 1.90 - 2.22 (m, 9H), 4.90 (bs, 1H), 5.25 (d, J = 4.5 Hz, 1H), 6.13 (s, 1H), 7.33-7.37 (m, 1H), 7.78 (m, 1H), 8.44 (dd, J = 4.8 and 1.5 Hz, 1H), 8.59 (d, J = 1.5 Hz, 1H), 10.34 (s, 1H).13C (75 MHz, DMSO): 16.96, 18.96, 20.57, 25.10, 25.53, 26.03, 28.73, 28.92, 30.30, 31.45, 31.72, 32.68, 33.56, 34.40, 35.03, 36.22, 37.19, 47.10, 49.03, 50.26, 57.52, 70.17, 121.57, 123.80, 129.45, 132.58, 133.75, 139.27, 147.61, 148.23, 151.44, 169.48, 172.76. HRMS (ESI) for C33H47N2O4 (M + H+): calcd, 535.3536; found, 535.3540.

[0177] Synthesis of (3R,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15-dode cahydro-1H-cyclopenta[a]phenanthren-3-yl 10-(hydroxyamino)-10-oxodecanoate (19).

[0178] The compound (19) was obtained in 19.6% yield using compound (36) in a manner similar to that described for the synthesis of compound (16); mp: 102-103 °C. HPLC purity: - 96.13%.1H NMR (300 MHz, DMSO): δ 1.03 (s, 3H), 1.05 (s, 3H), 1.24 (bs, 12H), 1.41-1.54 (m, 6H), 1.60-1.89 (m, 7H), 1.94-2.22 (m, 7H), 4.90 (bs, 1H), 5.25 (d, J = 5.1 Hz, 1H), 6.13 (bs, 1H), 7.38 (m, 1H), 7.78 (m, 1H), 8.44 (dd, J = 4.8 and 1.5 Hz, 1H), 8.59 (d, J = 1.8 Hz, 1H), 10.34 (s, 1H).13C (75 MHz, Acetone): 15.97, 18.29, 20.43, 25.04, 25.32, 25.91,28.76, 28.92, 29.11, 29.41, 30.31, 31.37, 31.41, 32.33, 33.52, 34.24, 35.17, 36.10, 37.06, 47.15, 50.55, 57.73, 69.89, 121.43, 123.07, 128.89, 132.73, 133.43, 139.10, 147.69, 147.82, 151.87, 169.75, 172.17. HRMS (ESI) for C34H49N2O4(M + H+): calcd, 549.3692; found, 549.3693.

[0179] Examples 9-12: Synthesis of Compounds (20) to (23)

[0180] Synthesis of (3S,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)- 2,3,4,7,8,9,10,11,12,13,14,15-dodecahydro-1H-cyclopenta[a]phenanthren-3-ylmethanesulfon ate (37)

[0181] A solution of compound (8) in pyridine was treated with methane sulfonyl chloride and stirred at room temperature for 3 hours. The reaction mixture was quenched with water and extracted with EtOAc (50 mL x 3). The combined organic layer was dried over anhydrous MgSO4, concentrated under reduced pressure, and was purified by silica gel chromatography (EtOAc: Hexane 2.5: 7.5) to give compound (37) in 84 % yield;1H NMR (300 MHz, CDCl3): d 1.11 - 1.12 (m, 6 H), 1.64-1.76 (m, 4 H), 1.79 – 1.98 (m, 6 H), 2.06 -2.19 (m, 4 H), 2.40 (m, 1 H), 2.55-2.58 (m, 2 H), 3.04 (s, 3 H), 4.56 (m, 1 H), 5.49 (d, J = 4.8 Hz, 1 H), 6.39 (s, 1 H), 7.88 (s, 1 H), 8.37 (d, J = 7.5 Hz, 1 H), 8.64 (s, 1 H), 8.75 (s, 1 H).

[0182] Synthesis of 3-((3S,8R,9S,10R,13S,14S)-3-azido-10,13-dimethyl2,3,4,7,8,9,10,11,12,13,14,15-dodecahydr o-1H cyclopenta[a]phenanthren-17-yl)pyridine (38).

[0183] A solution of compound (37) (420 mg, 0.001 mol) in DMF (10 mL) was treated with sodium azide (195 mg, 0.003 mol carefully. The mixture was stirred and heated to 80 °C for 3 hours. The mixture was cooled to 25 °C and quenched with water and extracted with EtOAc (50 mL x 3). The combined organic layer was dried over anhydrous MgSO4, concentrated under reduced pressure and was purified by silica gel chromatography (EtOAc : Hexane: 2:8) to give compound (38) in 84 % yield; NMR (300 MHz, DMSO): d 1.03 - 1.05 (s, 6 H), 1.39-1.81 (m, 11 H), 2.02-2.14 (m, 4 H), 2.19-2.23 (m, 2 H), 4.01 (m, 1 H), 5.37(d, J = 5.1 Hz, 1 H), 6.13 (m, 1 H), 7.35 (m, 1 H), 7.80 (m, 1 H), 8.44 (dd, J= 1.5 Hz & 4.7 Hz, 1 H), 8.60 (m, 1 H).

[0184] Synthesis of (3R,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)- 2,3,4,7,8,9,10,11,12,13,14,15-dodecahydro-1H-cyclopenta[a]phenanthren-3-amine (10).

[0185] To a solution of compound (38) in methanol (5 mL) and THF (3 mL) was added PPh (296 mg 0.00113 mol). The mixture was stirred and heated to 50 °C overnight. The reaction mixture was cooled to 25 °C and the pH was adjusted to 1 with 2 N aqueous solution, extracted with DCM (50 mL). The pH of the aqueous solution was adjusted to 9 with sat. NaHCO and extracted with DCM (50 mL x 2). The organic layers were combined and dried over anhydrous MgSO4, concentrated under reduced pressure and purified by silica gel chromatography (DCM: MeOH 93:7) to give compound (10) in 84 % yield;NMR (300 MHz, DMSO): d 0.99 - 1.00 (s, 6 H), 1.11 (m, 1 H), 1.26 (m, 2 H), 1.34-1.40 (m, 2 H), 1.47-1.51 (m, 3 H), 1.53 - 1.68 (m, 4 H), 1.80 (m, 1 H), 1.95-2.09 (m, 4 H), 2.21 (m, 1 H), 3.05 (s, 1 H), 5.27 (d, J= 2.7 Hz, 1 H), 6.09 (m, 1 H), 7.32 (m, 1 H), 7.74 (m, 1 H), 8.41 (m, 1 H), 8.57 (s, 1 H).

[0186] Synthesis of methyl 7-(((3R,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)- 2,3,4,7,8,9,10,11,12,13,14,15-dodecahydro-1H-cyclopenta[a]phenanthren-3-yl)amino)-7- oxoheptanoate (39)

[0187] A mixture of compound (10) (350 mg, 0.00122 mol), EDC.HCl (467mg, 0.0024 mol), HOBt (250 mg, 0.00185 mol), DIPEA (0.46 ml, 0.003 mol) in DMF (3 mL) and 7-methoxy-7-oxoheptanoic acid (230 mg, 1.7 mmol) was stirred at room temperature for 5h. The reaction mixture was quenched with water and extracted with EtOAc (50 mL x 3). The combined organic layer was dried over anhydrous MgSO4, concentrated under reduced pressure and was purified by silica gel chromatography (EtOAc: Hexane: 2:8) to give compound (39) in 84 % yield;1H NMR (300 MHz, CD3OD): δ 1.05 (s, 3H), 1.28 (s, 3H), 1.26 -1.44 (m, 12H), 1.73-2.05 (m, 7H), 2.12-2.40 (m, 11H), 3.61-3.70 (m, 1H), 4.21 (q, J =7.2 Hz, 2H), 5.63 (d, J = 4.6 Hz, 1H), 6.26 (bs, 1H), 7.50 (dd, J = 7.8 and 4.8 Hz, 1H), 7.97 (m, 1H), 8.55 (d, J = 3.9 Hz, 1H), 8.66 (s, 1H).

[0188] Synthesis of methyl 8-(((3R,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)- 2,3,4,7,8,9,10,11,12,13,14,15-dodecahydro-1H-cyclopenta[a]phenanthren-3-yl)amino)-8- oxooctanoate (40).

[0189] The title compound (40) was synthesized using compound (10) and 8-methoxy-8-oxooctanoic acid in a manner similar to that described for compound (39) in 72 % yield;1H NMR (300 MHz, CDCl3): d 1.03 (s, 3 H), 1.06 (s, 3 H), 1.10-1.16 (m, 2 H), 1.29-1.32 (m, 2 H), 1.43-1.49 (m, 2 H), 1.58-1.59 (m, 7 H), 1.66-1.72 (m, 7 H), 1.74-1.83 (m, 4 H), 1.92 (m, 1 H), 2.03-2.06 (m, 3 H), 2.09-2.14 (m, 2 H), 2.26 (m, 1 H), 2.58 (m, 1 H), 4.14 (m, 1 H), 5.41 (s, 1 H), 5.99 (s, 1 H), 7.23 (s, 1 H), 7.64 (d, J = 7.6 Hz, 1 H), 8.46 (s, 1 H), 8.62 (s, 1 H).

[0190] Synthesis of methyl 9-(((3R,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)- 2,3,4,7,8,9,10,11,12,13,14,15-dodecahydro-1H-cyclopenta[a]phenanthren-3-yl)amino)-9- oxononanoate (41).

[0191] The title compound (41) was synthesized using compound (10) and 9- methoxy-9-oxononanoic acid in a manner similar to that described for compound (39) in 76 % yield;1H NMR (300 MHz, CD3OD): d 1.07 (s, 3 H), 1.10 (s, 3 H), 1.21-1.28 (m, 2 H), 1.37 (m, 1 H), 1.44-1.52 (m, 2 H), 1.58-1.62 (m, 7 H), 1.63-1.69 (m, 5 H), 1.73-1.80 (m, 3 H), 2.05-2.13 (m, 7 H), 2.15 (t, J = 7.5 Hz, 2 H), 2.27 (m, 1 H), 2.56 (m, 1 H), 3.60 (s, 3 H), 4.00 (s, 1 H), 5.36 (d, J = 5 Hz, 1 H), 6.05 (s, 1 H), 7.34-7.36 (m, 1 H), 7.80 (d, J= 7.9 Hz, 1 H), 8.35 (d, J = 4.2 Hz, 1 H), 8.51 (s, 1 H).

[0192] Synthesis of methyl 10-(((3R,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)- 2,3,4,7,8,9,10,11,12,13,14,15-dodecahydro-1H-cyclopenta[a]phenanthren-3-yl)amino)- 10-oxodecanoate (42).

[0193] The title compound (42) was synthesized using compound (10) and 10- methoxy-10-oxononanoic acid in a manner similar to that described for compound (39) in 73 % yield;1H NMR (300 MHz, CDCl3): d 1.04 (s, 3 H), 1.07 (s, 3 H), 1.28 (bs, 10 H), 1.59 (bs, 7 H), 1.75 - 1.80 (m, 8 H), 2.05 - 2.14 (m, 5 H), 2.25 - 2.28 (m, 3 H), 3.64 (s, 3 H), 4.14 (s, 1 H), 5.41 (s, 1 H), 6.02 (s, 1 H), 7.26 (s, 1 H), 7.70 (d, J = 7.5 Hz, 1 H), 8.46 (s, 1 H), 8.62 (s, 1 H).

[0194] Synthesis of N1-((3R,10R,13S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9, 10,11,12, 13, 14,15-dodecahydro-1H-cyclopenta[a]phenanthren-3-yl)-N7-hydroxyheptanediamide (20).

[0195] To a solution of compound (39) (150 mg) in dioxane (4ml), LiOH (2 ml, 1M, aq) was added, and the reaction mixture was stirred at room temperature for 4 h. The reaction mixture was then acidified using HCl (3N) and white precipitates (0.16g) were filtered. To the solution of precipitates (0.16g, 0.00032 mol) in DMF (4 ml), EDC.HCl (0.125 g, 0.00065 mol), HOBt (66 mg, 0.00044 mol) and DIPEA (0.14ml, 0.00081 mol) were added and stirred at room temperature for 30 min before adding NH2OTHP (57mg, 0.48 mmol). After being stirred for a further 5 h, the reaction mixture was quenched with water and extracted with EtOAc (50 mL x 3). The combined organic layer was dried over anhydrous MgSO4, concentrated under reduced pressure, and purified by silica gel chromatography (EtOAc: Hexane: 4:6) to obtain a solid residue. To a solution of residue (120 mg) in CH3OH (4mL), 10% TFA (aq) (1mL) was added and the reaction was stirred at room temperature for 5 h. The reaction mixture was concentrated under reduced pressure to give white precipitates. The precipitates were recrystallized from CH3OH to afford the title compound (20) in 62% yield; mp: 90-91 °C; HPLC purity: - 96.11%1H NMR (300 MHz, CD3OD): d 1.08 (s, 3 H), 1.10 (s, 3 H), 1.18 -1.23 (m, 2 H), 1.31-1.35 (m, 3 H), 1.48 (m, 1 H), 1.55-1.60 (m, 6 H), 1.63-1.74 (m, 4 H), 1.76-1.83 (m, 2 H), 2.05-2.09 (m, 5 H), 2.13-2.21 (m, 2 H), 2.29 (m, 1 H), 2.56 (m, 1 H), 4.02 (s, 1 H), 5.38 (s, 1 H), 6.08 (s, 1 H), 7.36 (dd, J = 4.9 Hz & 8 Hz, 1 H), 7.83 (d, J =7.9 Hz, 1 H), 8.36 (s, 1 H), 8.52 (s, 1 H).13C NMR (75 MHz, CD3OD): 13.11, 15.63, 18.02,19.52, 20.33, 25.02, 25.44, 25.58, 28.22, 28.28, 30.28, 31.27, 31.39, 32.21, 33.39, 35.09, 35.26, 36.19, 37.11, 46.06, 120.91, 122.51, 123.72, 129.74, 134.81, 138.78, 141.03, 146.40, 146.60, 151.32, 171.47, 171.61, 174.22. HRMS (ESI) for C31H44N3O3 (M + H+): calcd, 506.3383; found, 506.3385

[0196] Synthesis of N1-((3R,10R,13S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9, 10,11,12,13, 14,15-dodecahydro-1H-cyclopenta[a]phenanthren-3-yl)-N8-hydroxyoctanediamide (21).

[0197] The title compound (21) was synthesized using compound (40) in a manner similar to that described for compound (20) in 61 % yield; mp: 98-99 °C; HPLC purity: - 95.66 %.1H NMR (300 MHz, CD3OD): d 1.09 (s, 3 H), 1.12 (s, 3 H), 1.23 (m, 1 H), 1.29-1.40 (m, 6 H), 1.50 (m, 1 H), 1.57-1.64 (m, 5 H), 1.68-1.75 (m, 4 H), 1.79 - 1.82 (m, 2 H), 2.06 - 2.15 (m, 6 H), 2.17 - 2.22 (m, 2 H), 2.30 (m, 1 H), 2.58 (m, 1 H), 4.03 (bs, 1 H), 5.39 (d, J = 5.1 Hz, 1 H), 6.09 (m, 1 H), 7.37 (dd, J = 4.8 Hz, and 7.8 Hz, 1 H), 7.84 (m, 1 H), 8.38 (d, J = 4.5 Hz, 1 H), 8.53 (s, 1 H).13C NMR (75 MHz, CD3OD): 15.64, 18.03, 20.34, 25.26,25.56, 25.74, 28.46, 30.29, 31.28, 31.38, 33.41, 35.10, 35.44, 36.20, 37.12, 46.07, 50.37, 57.73, 120.90, 122.50, 123.61, 129.48, 129.62, 133.62, 134.64, 138.80, 141.04, 146.78, 151.38, 171.57, 174.38. HRMS (ESI) for C32H46N3O3(M + H+): calcd, 520.3539; found, 520.3539

[0198] Synthesis of N1-((3R,10R,13S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13, 14,15-dodecahydro-1H-cyclopenta[a]phenanthren-3-yl)-N9-hydroxynonanediamide (22).

[0199] The title compound (22) was synthesized using compound (41) in a manner similar to that described for compound (20) in 65 % yield; mp: 101-102° C; HPLC purity: - 95.85%1H NMR (300 MHz, CD3OD): d 1.07 (s, 3 H), 1.10 (s, 3 H), 1.20-1.27 (m, 2 H), 1.36 (m, 1 H), 1.45-1.52 (m, 2 H), 1.58-1.62 (m, 7 H), 1.63-1.69 (m, 5 H), 1.73- 1.80 (m, 3 H),2.04-2.14 (m, 7 H), 2.19 (t, J = 7.7 Hz, 2 H), 2.29 (m, 1 H), 2.56 (m, 1 H), 4.01 (s, 1 H), 5.37 (d, J = 5.1 Hz, 1 H), 6.07 (s, 1 H), 7.36 (m, 1 H), 7.82 (d, J = 8 Hz, 1 H), 8.36 (d, J = 4.3 Hz, 1 H), 8.52 (s, 1 H).13C NMR (75 MHz, CD3OD): 15.68, 18.07, 20.36, 25.36, 25.59, 25.86, 28.70, 28.74, 30.28, 31.29, 31.40, 33.43, 35.12, 35.51, 36.22, 37.12, 46.07, 50.39, 57.75, 120.90, 123.65, 133.60, 141.05, 146.60, 146.80, 151.37, 171.55, 174.35, HRMS (ESI) for C33H48N3O3(M + H +): calcd, 534.3696; found, 534.3696.

[0200] Synthesis of N1-((3R,10R,13S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13, 14,15-dodecahydro-1H-cyclopenta[a]phenanthren-3-yl)-N10-hydroxydecanediamide (23).

[0201] The title compound (23) was synthesized using compound (42) in a manner similar to that described for compound (20) in 64 % yield; mp: 127-128 °C; HPLC purity: - 97.89%.1H NMR (300 MHz, CD3OD): d 1.06 (s, 3 H), 1.09 (s, 3 H), 1.29 (bs, 9 H), 1.57-1.59 (m, 5 H), 1.65 - 1.72 (m, 4 H), 1.75-1.82 (m, 2 H), 2.02-2.07 (m, 6 H), 2.17 (t, J = 7 Hz, 2 H), 2.27 (m, 1 H), 2.55 (m, 1 H), 4.00 (s, 1 H), 5.36 (d, J = 5.1 Hz, 1 H), 6.06 (s, 1 H), 7.34(dd, J = 4.9 Hz, & 7.7 Hz, 1 H), 7.81 (d, J = 8 Hz, 1 H), 8.35 (d, J = 4 Hz, 1 H), 8.50 (s, 1 H).13C NMR (75 MHz, CD3OD): 15.60, 18.00, 20.33, 25.38, 25.53, 25.89, 28.77, 28.95, 33.43, 35.13, 35.52, 36.16, 37.11, 50.46, 57.80, 122.48, 123.59, 129.62, 133.62, 134.64, 138.85, 146.75, 151.40, 174.44 HRMS (ESI) for C34H50N3O3(M + H +): calcd, 548.3852; found, 548.3853.

[0202] Examples 13-16: Synthesis of Compounds (24) to (27)

[0203] Synthesis of (3R,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15-dode cahydro-1H-cyclopenta[a]phenanthren-3-yl methanesulfonate (43).

[0204] To accomplish the synthesis of compound (43), the intermediate (9) (3 g, 0.008 mol) was dissolved in pyridine, and methane sulfonyl chloride was added to the solution. The reaction mixture was stirred for 3 hours at room temperature and then quenched with water and extracted with EtOAc (50 ml x 3). The combined organic layers were dried overanhydrous MgSO4 and concentrated under reduced pressure. The residue was purified by silica gel chromatography (n-Hexane: EtOAc = 7.5:2.5) to give a solid product (43) in 70.8% yield;1H NMR (300 MHz, CDCl3): δ 1.08 (s, 3H), 1.11 (d, J = 3.6 Hz, 3H), 1.62-1.87 (m, 11 H), 2.01-2.48 (m, 6H), 3.03 (s, 3H), 5.04 (bs, 1H), 5.44 (s, 1H), 6.03 (bs, 1H), 7.25 (m, 1H), 7.68 (m, 1H), 8.49 (dd, J = 4.8 Hz and 1.8 Hz, 1H), 8.65 (s, 1H).

[0205] Synthesis of 3-((3R,8R,9S,10R,13S,14S)-3-azido-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15-dodecahydr o-1H-cyclopenta[a]phenanthren-17-yl) pyridine (44).

[0206] A mixture of compound (43) (2.6 g, 0.006 mol) and sodium azide (1.5 g, 0.023 mol) was dissolved in DMF. The reaction mixture was heated at 80ºC for 3 hours and then quenched with water and extracted with EtOAc (50 ml x 3). The combined organic layers were dried over anhydrous MgSO4and concentrated under reduced pressure. The residue was purified by silica gel chromatography (n-Hexane: EtOAc = 7.5:2.5) to give a solid product (44) in 87.2% yield;1H NMR (300 MHz, DMSO): δ 1.03-1.05 (m, 9H), 1.09-1.25 (m, 6H), 1.63-1.71 (m, 2H), 1.98-2.29 (m, 7H), 5.45 (s, 1H), 6.13 (s, 1H), 7.35(m, 1H), 7.78 (dd, J = 7.8 Hz and 1.6 Hz, 1H), 8.45 (dd, J = 4.5 Hz, J = 1.2 Hz, 1H), 8.60 (s, 1H).

[0207] Synthesis of (3S,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15-dode cahydro-1H-cyclopenta[a]phenanthren-3-amine (11).

[0208] For synthesis of title compound (11), the intermediate (44) (1.9g, 0.005 mol), triphenylphosphine (2g, 0.007 mol), was dissolved in methanol and THF in a ratio of 20 ml: 12 ml. The reaction mixture was heated at 50ºC for overnight, quenched with water and extracted with butanol (50 ml x 3). The combined organic layers were dried over anhydrous MgSO4and concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM: Methanol = 95:5) to give a solid product (11) in 60.8% yield;1NMR (300 MHz, DMSO): δ 1.02-1.07 (m, 7H), 1.24-1.44 (m, 3H), 1.52-1.70 (m, 6H),1.77-1.88 (m, 4H), 2.02-2.35 (m, 5H), 3.18 (s, 1H), 5.34 (s, 1H), 6.13 (s, 1H), 7.35 (dd, J = 7.8 Hz and 4.8 Hz, 1H), 7.77 (d, J = 7.8 Hz, 1H), 8.44 (bs, 1H), 8.60 (s, 1H).

[0209] Synthesis of ethyl 7-(((3S,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15-d odecahydro-1H-cyclopenta[a]phenanthren-3-yl) amino)-7-oxoheptanoate (45).

[0210] A mixture of compound (11) (100 mg, 0.0002 mol), 7-ethoxy-7-oxoheptanoic acid (80 mg, 0.0004 mol), EDC.HCl (88 mg, 0.0005 mol) and HOBt (58 mg, 0.0004 mol) in DMF (3 ml) was stirred at room temperature for 10 min before adding DIPEA (0.15 ml, 0.0007 mol). After being stirred for a further 4 h, the reaction mixture was quenched with water and extracted with EtOAc (50 ml x 3). The combined organic layers were dried over anhydrous MgSO4 and concentrated under reduced pressure. The residue was purified by silica gel chromatography (n-hexane: EtOAc = 6: 4) to give a solid product (45) in 97% yield;1H NMR (300 MHz, CD3OD): δ 1.02 (s, 3H), 1.23 (s, 3H), 1.24 -1.43 (m, 12H), 1.72-2.07 (m, 7H), 2.18-2.46 (m, 11H), 3.67-3.71 (m, 1H), 4.25 (q, J = 7.2 Hz, 2H), 5.66 (d, J = 4.6 Hz, 1H), 6.23 (bs, 1H), 7.52 (dd, J = 7.8 and 4.8 Hz, 1H), 7.99 (m, 1H), 8.53 (d, J = 3.9 Hz, 1H), 8.68 (s, 1H).

[0211] methyl 8-(((3S,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15-d odecahydro-1H-cyclopenta[a]phenanthren-3-yl) amino)-8-oxooctanoate (46).

[0212] The compound (46) was obtained in 84% yield using compound (11) and 8-methoxy-8-oxooctanoic acid in a manner similar to that described for the synthesis of compound (45);1H NMR (300 MHz, CD3OD): δ 1.10 (s, 3H), 1.12 (s, 3H), 1.20-1.38 (m, 10H), 1.61-1.81 (m, 11 H), 2.16 - 2.37 (m, 8H), 3.66 (m, 1H), 3.68 (s, 3H) 5.44 (d, J = 4.8 Hz, 1H), 6.12 (bs, 1H), 7.41 (dd, J = 7.8 and 4.8 Hz, 1H), 7.87 (dt, J = 7.8 and 1.6 Hz, 1H), 8.41 (d, J = 4.8 Hz, 1H), 8.56 (bs, 1H).

[0213] methyl 9-(((3S,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15-d odecahydro-1H-cyclopenta[a]phenanthren-3-yl) amino)-9-oxononanoate (47).

[0214] The title compound (47) was synthesized in 98% yield by using a mixture of compound (11) and 9- methoxy-9-oxononanoic acid in a manner similar to that described for the synthesis of compound (45);1H NMR (300 MHz, CD3OD): δ 1.11 (s, 3H), 1.13 (s, 3H), 1.31- 1.36 (m, 12H), 1.61-1.69 (m, 6H), 1.73-2.10 (m, 6H), 2.16-2.37 (m, 7H), 3.66 (m, 1H), 3.68 (s, 3H), 5.45 (d, J = 4.4 Hz, 1H), 6.12 (dd, J = 3.0 and 1.8 Hz, IH), 7.41 (dd, J = 8.1 and 1.8 Hz, 1H), 7.87 (m, 1H), 8.41 (d, J = 4.2 Hz, 1H), 8.56 (s, 1H).

[0215] methyl 10-(((3S,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15- dodecahydro-1H-cyclopenta[a]phenanthren-3-yl) amino)-10-oxodecanoate (48).

[0216] The compound (48) was obtained in 76 % yield using compound (11) and 10- methoxy-10-oxononanoic acid in a manner similar to that described for the synthesis of compound (45);1H NMR (300 MHz, CD3OD): δ 1.12 (s, 3H), 1.13 (s, 3H), 1.35 (m, 15 H), 1.61-1.78 (m, 12H), 2.07- 2.37 (m, 6H), 3.66 (m, 1H), 3.68 (s, 3H) 5.45 (d, J = 5.4 Hz, 1H), 6.12 (bs, 1H), 7.41(dd, J = 7.5 and 4.8 Hz, 1H), 7.87 (m, 1H), 8.41 (d, J = 4.8 Hz, 1H), 8.56 (bs, 1H).

[0217] Synthesis of N1-((3S,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15- dodecahydro-1H-cyclopenta[a]phenanthren-3-yl)-N7-hydroxyheptanediamide (24).

[0218] To a solution of compound (45) (160 mg, 0.0003 mol) in dioxane (4ml), LiOH (2 ml, 1Maq 0.0015 mol) was added and the reaction mixture was stirred at room temperature for 4 h. The reaction mixture was then acidified using HCl (3N), and white precipitates (0.145 g) were filtered. To the solution of precipitates (0.145 g, 0.0002 mol) in DMF (4 ml), EDC.HCl (91 mg, 0.0005 mol), HOBt (60 mg, 0.0004 mol), and DIPEA (0.15 ml, 0.0007 mol) wereadded and stirred at room temperature for 30 min before adding NH2OTHP (70 mg, 0.0005 mol). After being stirred for a further 5 h, the reaction mixture was quenched with water and extracted with EtOAc (50 mL x 3). The combined organic layer was dried over anhydrous MgSO4, concentrated under reduced pressure, and purified by silica gel chromatography (EtOAc: Hexane: 4:6) to obtain a solid residue. To a solution of residue (140 mg 0.0002 mol) in CH3OH (4mL), 10% TFA (aq) (1.5 mL) was added and the reaction was stirred at room temperature for 3 h. The reaction mixture was concentrated under reduced pressure to give white precipitates. The precipitates were recrystallized from CH3OH to afford the title compound (24) in 18.3% yield; mp: 95-96 °C; HPLC purity: - 95.59%1H NMR (300 MHz, CD3OD): δ 1.08 (s, 3H), 1.09 (s, 3H), 1.13-1.34 (m, 7H), 1.54-1.63 (m, 7H), 1.68-1.90 (m, 5H), 2.06 - 2.21 (m, 8H), 3.54 (m, 1H), 5.39 (d, J = 4.8 Hz, 1H), 6.24 (s, 1H), 7.65 (dd, J = 7.8 and 5.4 Hz, 1H), 8.18 (d, J = 8.4 Hz, 1H), 8.50 (bs, 1H), 8.65 (bs, 1H).13C (75 MHz, CD3OD): 15.43, 18.29, 20.47, 24.94, 25.23, 28.16, 28.22, 30.27, 31.10, 31.50, 32.14, 34.78, 35.55, 36.55, 37.74, 38.43, 47.14, 49.59, 50.48, 57.56, 120.78, 125.20, 132.27, 135.22, 138.73, 141.01, 142.74, 143.05, 149.87, 171.45, 173.76. HRMS (ESI) for C31H44N3O3(M + H+): calcd, 506.3383; found, 506.3383.

[0219] Synthesis of N1-((3S,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15- dodecahydro-1H-cyclopenta[a]phenanthren-3-yl)-N8-hydroxyoctanediamide (25)

[0220] Compound (25) was obtained in 19.16 % yield from compound (46) in a manner similar to that described for the synthesis of compound (24); mp: 104-105 °C HPLC purity: - 98.31 %1H NMR (300 MHz, CD3OD): δ 1.11 (s, 3H), 1.13 (s, 3H), 1.18-1.55 (m, 8H), 1.63-1.82 (m, 11H), 1.94-2.26 (m, 10H), 3.58 (m, 1H), 5.44 (d, J = 4.2 Hz, 1H), 6.13 (bs, 1H), 7.44 (bs, 1H), 7.92 (m, 1H), 8.44 (bs, 1H), 8.58 (bs, 1H).13C (75 MHz, CD3OD): 15.57, 18.31, 20.51, 24.16, 25.49, 28.25, 28.37, 28.40, 30.34, 31.16, 31.33, 32.25, 35.08, 35.71, 36.56, 37.75, 38.46, 47.10, 49.59, 50.55, 57.61, 120.87, 123.56, 129.56, 134.59, 138.78,140.99, 146.51, 146.68, 151.35, 171.44, 173.88. HRMS (ESI) for C32H46N3O3 (M + H+): calcd, 520.3539; found, 520.3544.

[0221] Synthesis of N1-((3S,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15- dodecahydro-1H-cyclopenta[a]phenanthren-3-yl)-N9-hydroxynonanediamide (26).

[0222] Compound (26) was obtained in 24.7 % yield from compound (47) in a manner similar to that described for the synthesis of compound (24);mp: 97-98 °C HPLC purity: - 96.68 %1H NMR (300 MHz, DMSO): δ 1.03 (s, 3H), 1.06 (s, 3H), 1.09-1.25 (m, 13H), 1.45-1.48 (m, 6H), 1.94 -2.25 (m, 9H), 2.10-2.25 (m, 3H), 3.34 (m, 1H), 5.35 (d, J = 4.2 Hz, 1H), 6.13 (bs, 1H), 7.35 (m, 1H), 7.69 (d, J = 7.8 Hz, 1H), 7.78 (m, 1H), 8.45 (dd, J = 4.8 and 1.8 Hz, 1H), 8.60 (d, J = 1.5 Hz, 1H), 8.65 (s, 1H), 10.33 (bs, 1H).13C (75 MHz, CD3OD): 15.57, 16.87, 17.91, 18.31, 19.81, 20.50, 20.74, 22.20, 25.24, 25.58, 28.53, 30.34, 31.33, 32.29, 35.05, 35.78, 36.56, 37.75, 38.46, 50.56, 57.62, 69.64, 120.86, 123.53, 129.57, 133.57, 134.63, 140.99, 146.49, 146.67, 151.33, 171.59, 173.95. HRMS (ESI) for C33H48N3O3 (M + H+): calcd, 534.3696; found, 534.3696.

[0223] Synthesis of N1-((3S,8R,9S,10R,13S,14S)-10,13-dimethyl-17-(pyridin-3-yl)-2,3,4,7,8,9,10,11,12,13,14,15- dodecahydro-1H-cyclopenta[a]phenanthren-3-yl)-N10-hydroxydecanediamide (27).

[0224] Compound (27) was obtained in 30.2% yield from compound (48) in a manner similar to that described for the synthesis of compound (24); mp: 96-97 °C HPLC purity: - 98.08 %.1H NMR (300 MHz, CD3OD): δ 1.05 (s, 3H), 1.14 (s, 3H). 1.18-1.56 (m, 13H), 1.60-1.81 (m, 10H), 1.92-2.26 (m, 10H), 3.60 (m, 1H), 5.45 (d, J = 4.5 Hz, 1H), 6.32 (dd, J = 3.3 and 1.8 Hz, 1H), 7.74 (t, J = 6.0 Hz, 1H), 8.28 (d, J = 7.8 Hz, 1H), 8.58 (bs, 1H), 8.73 (bs, 1H).13C (75 MHz, CD3OD): 15.41, 18.27, 20.47, 25.27, 25.62, 28.24, 28.63, 28.71, 28.73, 28.78, 28.89, 30.28, 31.09, 31.47, 32.33, 34.79, 35.79, 36.50, 37.73, 38.44, 49.57, 50.48,57.57, 120.57, 120.81, 129.42, 132.00, 138.33, 140.99, 143.10, 143.36, 150.04, 171.59, 174.00 HRMS (ESI) for C34H50N3O3 (M + H+): calcd, 548.3852; found, 548.3855.

[0225] Example 17: In vitro cytotoxicity studies

[0226] Initially, all the synthesized compounds were evaluated for their cell growth inhibitory effects against Pt#3 human cancer cell lines. Abiraterone (8) was employed as the standard in this study, which provided key insights into the impact of structural variation on cytotoxicity (Table 1). Careful observation of the results revealed that abiraterone exhibited weak antiproliferative effects against the cell lines; however, attaching the chemical architecture of abiraterone to hydroxamic acid functionality via an ester bond with a long methylene chain (five –CH2- units, compound 12) enhanced the cell growth inhibitory potential of the resulting structural framework. Compound 12 inhibited the Pt#3 cell lines with an IC50value of 10.59 µM. Nevertheless, homologating the alkyl chain did not yield more positive results, as compounds (13-15) bearing longer methylene chains (–CH2- units - 6, 7, and 8) seem to have similar or less antiproliferative effects. Further, evaluations were conducted in pursuit of assessing the impact of the inversion of the configuration at position 3. It seems that compounds (16 – 19) (long alkyl chain hydroxamic acids with the nverted configuration at position 3) failed to elicit higher antitumor effects. Thus, it was deduced that the inversion of configuration at position 3 may not be a key point of enhancing the activity as compounds (16-19) were more or less potent as compared to Abiraterone (8) in inhibiting the Pt#3 cells. Further assessments were aimed at pinpointing a more potent CYP17A1 inhibitor than compound 12 and the strategy of isosteric replacement of O with NH (both configurations – retained as well as inverted, position 3) in the structural template of compound 12 was explored. It appears that none of the compounds (20-27) could outshine their counterpart compound 12 in terms of enhanced cell growth inhibition. However, some interesting insights were still gained while profiling compounds (20-27) as cell growth inhibitors. Notably, the concomitant replacement of O with NH and inversion of configurationled to the regeneration of activity and varied preferability of the SRP for the alkyl chain length. Unlike compound 12 bearing a five methylene chain, the inverted configuration (position 3 and NH-bearing adducts) demonstrated moderate cytotoxic effects with a chain of 8 –CH2-units (compound 23, IC50 = 21.48 µM). Also, the evaluation of the retained configuration of NH-bearing adducts yielded encouraging results, as compounds (24-27) were found to be better than their counterparts (20-23) at inhibiting the Pt#3 cell lines. Specifically, compound 25 with an amide bond in place of an ester bond, and the retained configuration (position 3) exerted significant cell growth inhibitory effects against the employed cell lines (IC50= 13.89 µM). Again, generalization in the context of the length of the methylene chain could not be made as the presence of six methylene chains within the structural framework of compound 25 was found to be the optimal linker length for SRP 11. Despite promising cell growth inhibitory effects, hydroxamic acid 25 seemed not to be superior to scaffold 12. Overall, it was deduced that an ester linkage and retention of configuration at position 3 (S configuration as in Abiraterone) might be crucial for antiproliferative effects. To sum up, compound 12 was identified as a preferred compound and was applied in further mechanistic studies.

[0227] Table 1 shows cell growth inhibitory effects of abiraterone (8) and compounds (12 -27):Table 1

[0228] Example 18: In-vitro mechanistic studies (CYP17A1 and HDAC inhibition)

[0229] Given the promising cell growth inhibitory effects of compound 12, attempts were made to investigate whether the cell growth inhibitory effects of hydroxamic acid 12 are driven by dual modulation of CYP17A1 and HDAC. For the evaluation of CYP17A1 inhibitory activity of compound 12, DHEA levels were measured. Abiraterone (8) was employed as a standard for the assay. Delightfully, the most potent cell growth inhibitor 12 exhibited a magnificent CYP17A1 profile with an IC50value of 0.284 µM. Notably, hydroxamic acid 12 demonstrated a more pronounced CYP17A1 inhibitory activity than the standard compound, Abiraterone (8). Subsequently, the assessment of CYP17A1 inhibitory activity and the HDAC inhibitory effects of compound 12 against the HDAC6 isoform was evaluated. The outcome of the assay (CYP17A1 and HDAC isoform inhibition) is depicted in Table 2, which clearly indicates that compound 12 inhibits HDAC6 with an IC50 value of 0.60 µM. Collectively, the results presented in Table 2 demonstrate the dual inhibitory potential of compound 12. Given the results of CYP17A1 / HDAC6 inhibition coupled with the literature precedents that advocate for the favorable trends with CYP17A1 and HDAC6 isoform targeting in GBM as their overexpression promotes GBM cells proliferation, it was assumed that the anti-glioma effects of compound 12 stem from dual CYP17A1 and HDAC6 inhibition.aND, not determined; Empty cells indicate no inhibition or compound activity that could not be fit to an IC50 curve. Table 2

[0230] In comparison in anti-GBM activity, compound 12 exhibited the higher activity in suppressing cell viability in TMZ-resistant U87MG-R and A172-R cells (see, Fig. 1A). The results of the in-vitro CYP17A1 and HDAC6 inhibition were further rationalized with molecular modeling studies. The outcome of the docking studies reveals that compound 12 was well accommodated in the active site of CYP17A1 and HDAC6, as shown in Figs. 1B and 1C. For CYP17A1, a computational analysis revealed that the wild-type protein exhibited a substantially lower docking energy of -173.908 compared to its mutant form (G297A / G301A), which had a docking energy of -132.436. This substantial difference in docking energy strongly suggests that the wild-type CYP17A1 forms a more robust and favorable interaction with compound 12 than its mutant counterpart. Moreover, when examining the hydrogen bond contributions, it was observed that the wild-type protein had a higher hydrogen bond contribution of 41.512, whereas the mutant form showed a reduced hydrogen bond contribution of -11.436. This divergence in hydrogen bonding energy indicates that the mutations (G297A / G301A) might alter the binding site, thereby diminishing the protein’s ability to form hydrogen bonds with compound 12.

[0231] Turning the attention to HDAC6, the computational analysis indicated a similar trend. The wild-type HDAC6 displayed a lower docking energy of -145.897, while the mutant form (H610F / C621V) had a higher docking energy of -107.162. This discrepancy in docking energy emphasizes the stronger interaction between compound 12 and the wild-type HDAC6. Furthermore, when examining hydrogen bond contributions, it was noted that thewild-type protein exhibited a greater hydrogen bond contribution of -34.5149, whereas the mutant form displayed a reduced hydrogen bond contribution of -16.1599. This disparity implies that mutations in HDAC6 have indeed affected the binding affinity, resulting in altered hydrogen bond interactions and weaker binding forces in the mutant form.

[0232] To sum up, these molecular modeling results provide detailed evidence of how mutations in CYP17A1 and HDAC6 can significantly influence their binding affinities with compound 12. The lower docking energies and altered hydrogen bonding interactions in the wild-type proteins underscore the importance of considering protein mutations in drug development strategies. This understanding not only validates the efficacy of compound 12 but also underscores the critical role of comprehending how mutations impact drug-protein interactions, ultimately guiding the optimization of drug development for more effective treatments, particularly in the context of proteins with mutant variants.

[0233] Example 19: In-vitro cytotoxicity against TMZ-resistant cell lines

[0234] Further, compound 12 was assessed for the cell growth inhibitory effects against TMZ-resistant cells (T98G, A172-R, Pt#3-R, and U87MG-R). Encouragingly, hydroxamic acid 12 manifested an impressive in-vitro antitumor profile against GBM cells as well as TMZ-resistant GBM cells. Notably, compound 12 elicited more pronounced cell growth inhibitory effects against TMZ-resistant cell lines viz U87MG-R with IC50values of 14.26 µM in comparison to GBM cell lines (U87MG). Also, compound 12 was almost equipotent against Pt#3-R (TMZ resistant cell lines, IC50 = 9.76 µM) and Pt#3 cell lines (GBM cells, IC50 = 9.49 µM) (Table 3) as well as against A172R (IC50= 12.23 µM) and A172 (IC50= 11.49 µM). It is important to note that 12 did not exert any cytotoxicity against normal astrocytes (IC50 = 281.6 µM) (see, Fig. 2A) and this observation is indicative of the selective effects of the compound towards cancer cell lines.p p Table 3

[0235] To validate whether the suppressive effect of compound 12 is mediated through CYP17A1 and HDAC6, 2 kinds of U87MG cells were generated, one with overexpression of CYP17A1 and HDAC6 (through gene transfection) and the other with gene silencing (through CRISPR-mediated knockout of HDAC6 and siRNA-mediated CYP17A1 knockdown). (Fig. 2B). The results depicted in Fig. 2C clearly indicate that, as compared with the control group (HA / DDK), overexpression of CYP17A1 and HDAC6 decreased IC50 from 15.22 to 10.65 µM and the silencing of HDAC6 and CYP17A1 increased IC50from 15.22 to 37.06 µM (Fig. 2C). In addition, IC50 was decreased from 37.06 to 27.62 µM by gene overexpression in gene-silent U87MG cells (Fig.2C), indicating that 12 suppresses GBM in the CYP17A1- and HDAC6-dependent manner. Moreover, compound 12, not abiraterone, significantly induced apoptosis at 10 µM in TMZ-resistant U87MG-R and Pt#3-R (Figs.2D-2E)

[0236] Example 20: Induction of DNA damage and increase in ROS accumulation

[0237] To understand the mechanism underlying compound 12-mediated GBM suppression, RNA-seq was also performed to identify the compound 12-influenced genes. In Figs.3A-3B, genes downregulated by compound 12 were functionally annotated.

[0238] In particular, compound 12 exerted the most significant effect on the DNA metabolic process, including DNA damage response. Spurred by the aforementioned findings,the inventors further investigated whether DNA damage was initiated by compound 12. As shown in Figure 3C, DNA double-strand break characterized by H2Ax was clearly induced by compound 12. Notably, this effect was not demonstrated by Abiraterone (Fig.3C).

[0239] Further, in addition to decreasing DNA repair protein Rad51 expression, compound 12 also manifested the ability to decrease the levels of antioxidant proteins, including sodium dismutase (SOD) 1 / 2 and glutathione peroxidase (GPX) 1 / 4 (Fig. 3D). Additionally, compound 12 enhanced ROS production and induced ROS leakage from mitochondria (Fig.3E-3F).

[0240] Example 21: Suppression of the expression of recurrence-associated genes in GBM

[0241] The assessment endeavor was further directed towards the elucidation of the ability of compound 12 to suppress gene expression associated with TMZ resistance and tumor recurrence. As shown in Fig. 4A, hydroxamic acid 12 demonstrated the potential to decrease the expression of genes that are upregulated in TMZ-resistant Pt#3 cells. Notably, among the genes downregulated by compound 12 (Fig. 4A), the expression of seven genes (CDH23, LRMP, LCP1, IFI44L, SLC25A27, NPY1R, and SLITRK1) was found to be elevated after recurrence (Fig. 4B). Also, the study results validated the ability of compound 12 to decrease the expression of IFI44L, LCP1, SLC25A27, SLITRK1, and NPY1R in a dose-dependent manner (Fig.4C).

[0242] Example 22: Inhibition of the growth of TMZ-resistant GBM without inducing significant adverse effects in vivo

[0243] In-vivo study to assess the tumor growth inhibitory potential of compound 12 was conducted in the xenograft model (U87MG-R-induced tumors). Delightfully, compound 12 at the dose of 20 mg / kg significantly suppressed the growth of U87MG-R-induced tumors (Fig. 5A). It is worth mentioning that compound 12, in comparison to abiraterone which failed tosuppress the tumor, elicited tumor growth inhibitory effects even at the dose of 5 mg / kg (Fig. 5B).

[0244] Further in-vivo investigations were performed in the orthotopic brain tumor mouse model (Pt#3-R cells injected into the brain). Encouragingly, administration (i.p) of 5 mg / kg of compound 12 significantly extended the survival period of experimental mice (Fig.5C). Also, tumor size was found to be decreased (Fig. 5D). Importantly, compound 12 exhibited low bio-toxicity as there was no decrease in the body weight of mice (Fig.5E).

[0245] Also, compound 12 at a high dose (100 mg / kg orally, once per week), as well as a low dose (10~20 mg / kg orally, 4 times per week), significantly extended the survival of experimental mice without decreasing the body weight. (Fig.5F-5H).

[0246] Spurred by the precedential claims regarding the candidature of CYP17A1 as a potential target for GBM, this medicinal chemistry campaign was designed to furnish dual CYP17A1 – HDAC inhibitors. To construct dual modulatory assemblages, the accommodative ability of the HDAC inhibitory template to recruit diverse pharmacophores was exploited, and the structural framework of Abiraterone was utilized as a CAP construct. Hydroxamic acid functionality was appended to the Abiraterone core via methylene chain linkers. In addition to the influence of linker length variation (homologation and truncation), the effect of the inversion of configuration at position 3 of the abiraterone framework and isosteric replacement of oxygen with NH on the activity was also studied. The results of the cellular activity revealed that ester functionality bearing five –CH2-units as a linker, retention of configuration, and oxygen functionality at position 3, were the optimized structural features. The correlation of CYP17A1 and HDAC6 inhibitory activity of the compound 12 with the cell growth inhibitory effects against the GBM cell lines revealed that the antitumor potential of compound 12 stems from its dual modulatory ability of CYP17A1 and HDAC6. Delightfully, TMZ-resistant cells (T98G, A172-R, Pt#3-R, and U87MG-R) were found to be equally sensitive to the exposure of compound 12, which indicates the ability of the dual inhibitor to overcome the TMZresistance. Further explorations underscored the effect of compound 12 on the DNA metabolic process, including DNA damage response as well as its ability to suppress gene expression associated with TMZ resistance and tumor recurrence. Encouragingly, the results of the in-vivo studies were overwhelmingly positive as compound 12 could substantially suppress the growth of U87MG-R-induced tumors bearing xenograft. Also, intraperitoneal administration of compound 12 (orthotopic brain tumor mouse model) significantly extended the survival period of experimental mice. Thus, these findings clearly ascertain that the striking in-vitro efficacy of the dual inhibitor 12 translated well to in-vivo efficacy. In a nut shell, compound 12, as well as the compounds of Formula (I), may be promising anti-GBM agents that can overcome TMZ resistance in glioblastoma.

[0247] Example 23: Inhibition of Abiraterone-resistant prostate tumor growth in vivo

[0248] In addition to brain cancer cell lines, it is believed that the compounds and derivatives disclosed herein would exhibit anti-tumor effect on other types of cancer. Thus, 22rv1 cells (1 ^ 106cells in 50 ŦL of DMEM), as an abiraterone-resistant prostate tumor model, was injected into the back of 8-week-old CAnN.Cg-Foxn1nu / CrlBltw (BALB / c nude) male mice. A month after transplantation, mice were administrated with abiraterone (Compound 8) or Compound 12 twice a week by intraperitoneal injection. Tumor size was calculated according to the formula: 1 / 2 x long side x (short side). Results are shown in Figs. 6A and 6B and Compound 12 exhibited superior effects, in particular over the control group (DMSO) and comparative group (Compound 8), on the inhibition of prostate cancer cell growth in the xenograft mice model.

[0249] A person of ordinary skill in the art of the subject disclosure should understand that variations and modifications may be made to the teaching and disclosure of the subject application without departing from the spirit and scope thereof. Based on the preceding contents, the subject application intends to cover any variations and modification thereof with the proviso that the variations or modifications fall within the scope as defined in the appended claims or their equivalents.

Claims

We claim:

1. A compound of Formula (I),or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof, wherein: X is O, NRaor S wherein Rais H or C1-5alkyl, preferably X is O or NH; R is H, C1-5alkyl, -C(=O)Rb, -C(=O)ORbor –C(=O)-NRbwherein Rbis H or C1-5alkyl, preferably R is H; and n is an integer ranging from 2 to 10, preferably 4 to 9, more preferably 5 to 8; wherein each of RA1, RA2, RA3 and RA4 substituent is independently halogen, hydroxy, C1-5alkyl or haloC1-5alkyl; q, r, s and t each independently represent an integer ranging from 0 to 3, provided that the sum of q, r, s and t ranges from 0 to 5. . The compound of claim 1, wherein Formula (I) is Formula (IA) or Formula (IB):wherein X, R, n, RA1, RA2, RA3, RA4, q, r, s and t are as defined in Claim 1.

3. The compound of any one of proceeding claims, wherein Formula (I) is Formula (I'):wherein R, n, RA1, RA2, RA3, RA4, q, r, s and t are as defined in Claim 1.

4. The compound of any one of proceeding claims, wherein Formula (I) is Formula (IA') or Formula (IB'):(IA'),(IB'), wherein R, n, RA1, RA2, RA3, RA4, q, r, s and t are as defined in Claim 1.

5. The compound of any one of proceeding claims, wherein Formula (I) is Formula (I"):wherein R, n, RA1, RA2, RA3, RA4, q, r, s and t are as defined in Claim 1.

6. The compound of any one of proceeding claims, wherein Formula (I) is Formula (IA") or Formula (IB"):wherein R, n, RA1, RA2, RA3, RA4, q, r, s and t are as defined in Claim 1.

7. The compound of any one of proceeding claims, wherein X is O, NRaor S wherein Rais H or C1-5alkyl, R is H or C1-5alkyl, and n is an integer ranging from 5 to 8.

8. The compound of any one of proceeding claims, wherein X is O, NRaor S wherein Rais H or C1-5alkyl, R is H and n is an integer ranging from 4 to 9.

9. The compound of any one of preceding claims, wherein R is H and n is an integer ranging from 5 to 8.

10. The compound of any one of preceding claims, wherein each of RA1, RA2, RA3and RA4is independently halogen, hydroxy, C1-5alkyl or haloC1-5alkyl; q, r, s and t each independently represent an integer ranging from 0 to 1.

11. The compound of any one of preceding claims, wherein the sum of q, r, s and t is 0.

12. The compound of any one of proceeding claims, which is selected from the group consisting of: Formula Structure Formula StructureFormula Structure Formula Structureor a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof.

13. The compound of any one of proceeding claims selected from the group consisting of:, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof.

14. A pharmaceutical composition comprising the compound or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or isomer thereof of any preceding claims, and a pharmaceutically acceptable carrier.

15. A pharmaceutical composition for use in treating and / or preventing a disease or disorder mediated by CYP17A1, HDAC or both in a subject, comprising the compound of any one of proceeding claims or a pharmaceutically acceptable salt, hydrate, solvate or prodrug of any of the foregoing to a subject in need thereof.

16. The pharmaceutical composition of claim 15, wherein the disease or disorder is a CYP17A1-HDAC6 mediated disease or disorder.

17. A pharmaceutical composition for use in treating and / or preventing a cancer in a subject, comprising the compound of any one of proceeding claims or a pharmaceutically acceptable salt, hydrate, solvate or prodrug of any of the foregoing to a subject in need thereof.

18. The method according to claim 17, wherein the cancers are hematological malignancies or solid tumors.

19. The pharmaceutical composition according to claim 17, wherein the cancer is brain cancer, prostate cancer, lymphoma, myeloma (such as multiple myeloma), leukemia, melanoma, lung cancer (such as small cell lung cancer and non-small cell lung carcinoma (NSCLC) (such as squamous cell carcinoma, adenocarcinoma and large cell carcinoma)), breast cancer (such as metastatic breast cancer), cervical cancer, colorectal cancer (CRC), cholangiocarcinoma, esophageal cancer, liver cancer, colon cancer, pancreatic cancer, stomach cancer and renal cancer.

20. The pharmaceutical composition of any one of claims 15 to 19, wherein the cancer is a brain cancer, prostate cancer, a metastatic brain cancer or a metastatic prostate cancer.

21. The pharmaceutical composition of claim 20, wherein the brain cancer is glioma or meningioma.

22. The pharmaceutical composition of claim 20, wherein the brain cancer is glioblastoma (GBM), medulloblastoma, or acoustic neuroma.

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