Neuroactive steroids and pharmaceutical compositions containing the same

Abstract

The present invention relates to novel neuroactive steroids (NAS). The present invention also relates to pharmaceutical compositions comprising the neuroactive steroids (NAS) and salts thereof. The pharmaceutical compositions can be used to prevent and / or treat CNS conditions or diseases associated with GABA regulation, such as depression, bipolar disorder, dementia, Huntington's disease, Parkinson's disease, and the like. The present invention also relates to methods for treating CNS disorders in subjects in need thereof.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of and priority to U.S. Provisional Application Serial No. 62 / 959,977, filed January 12, 2020, which is incorporated herein by reference in its entirety. Technical Field

[0003] The present disclosure relates to neuroactive steroids (NAS) and pharmaceutical compositions comprising the same. The present disclosure also relates to methods of using neuroactive steroids to treat central nervous system (CNS) disorders. Background Art

[0004] Neuroactive steroids (NAS), including neurosteroids (NS), are modulators of the gamma-aminobutyric acid (GABA) receptor complex (GRC) in the central nervous system (CNS). The primary target of NAS is the inhibitory GABA type A receptor (GABA A R), which helps regulate neuronal excitability and rapid mood changes. NAS can be produced de novo in the brain from cholesterol or by local metabolism of peripherally derived steroid precursors. Endogenous neurosteroids may include 5α-pregnane-3α-ol-20-one (allopregnanolone, also known as brexanolone) and 5α-pregnane-3α,21-diol-20-one (THDOC) (Majewska MD et al., Science. 232:1004–7, 1986 [PubMed:2422758]). Synthetic neuroactive steroids, such as alphaxalone, can also selectively enhance responses to GABA (Harrison NL et al., J Physiol (Lond). 346:42, 1984). Many CNS disorders (including a variety of behavioral states, such as anxiety levels, panic, stress reactions, epileptic seizures, sleep, alertness, and memory) can be linked to GABA. A R is functionally relevant and can be affected by NAS and its synthetic derivatives (Zorumski, CF. et al., Neurosci. Biobehavioral Rev. 37: 109-122, 2013. DOI: 10.1016 / j.neubiorev.2012.10.005).

[0005] Given its critical role in neuronal circuit function, GABA A GABA R is the target of many clinically relevant drugs. ... ARecently, the U.S. Food and Drug Administration (FDA, March 2019) approved a product for intravenous injection of brexanolone solution developed by Sage Therapeutics (Cambridge, MA, USA) and sold under its registered trademark For the treatment of postpartum depression (PPD), a serious and potentially life-threatening condition for which there is currently no specific drug therapy. However, ZULRESSO is inconvenient to use and requires administration to patients via continuous intravenous (IV) infusion lasting a total of approximately 60 hours (2.5 days). A new oral drug, GABA A A positive allosteric modulator of R, called SAGE-217, showed mixed results in reducing depressive symptoms after 14 days of administration in clinical trials. In addition, SAGE-217 showed more adverse events than placebo (Gunduz-Bruce, H., et al., N. Engl. JM ed. 381(10):903-911, September 2019; U.S. Patent No. 9,512,165 and PCT Publication No. WO2014 / 169833). Many new molecules have also been proposed, such as those disclosed in U.S. Patent No. 9,777,037 and U.S. Patent Publication No. 20180340005A1. However, their effectiveness in treating human CNS disorders is still unclear.

[0006] Therefore, it is necessary to regulate GABA A R function and better compounds for treating CNS disorders. Summary of the Invention

[0007] The present disclosure relates to neuroactive steroids (NAS) of formula (1):

[0008]

[0009] or one or more isomers thereof, deuterium-labeled variants thereof, or combinations thereof, wherein: R1 is independently H, D, substituted or unsubstituted C1-C10 alkyl, C1-C5 deuterated alkyl, substituted or unsubstituted C2-C10 alkenyl, substituted or unsubstituted C2-C10 alkynyl, substituted or unsubstituted C3-C10 cycloalkyl, substituted or unsubstituted C3-C10 cycloalkenyl, substituted or unsubstituted C3-C10 heterocycloalkyl, substituted or unsubstituted C3-C10 heterocycloalkenyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R2, R4, and R5 are each independently H, halogen, -CN, substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C2-C10 alkenyl, substituted or unsubstituted C2-C10 alkynyl, substituted or unsubstituted C3-C10 cycloalkyl, R3 is H, D, halogen, -CN, substituted or unsubstituted C1-C10 alkyl, -CD3, substituted or unsubstituted C2-C10 alkenyl, substituted or unsubstituted C2-C10 alkynyl, substituted or unsubstituted C3-C10 cycloalkyl, substituted or unsubstituted C3-C10 cycloalkenyl, substituted or unsubstituted C3-C10 heterocycloalkyl, substituted or unsubstituted C3-C10 heterocycloalkenyl, substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl; R6 is H or D; and m and n are each independently 0, 1, 2 or 3, with the proviso that at least one of m and n is not zero.

[0010] The present invention also relates to a pharmaceutical composition comprising a neuroactive steroid (NAS) of formula (1), one or more isomers thereof, deuterium-labeled variants thereof, a pharmaceutically acceptable salt thereof, or a combination thereof; and a pharmaceutically acceptable excipient; wherein: R1 is independently H, D, substituted or unsubstituted C1-C10 alkyl, C1-C5 deuterated alkyl, substituted or unsubstituted C2-C10 alkenyl, substituted or unsubstituted C2-C10 alkynyl, R2, R4 and R5 are each independently H, halogen, -CN, substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C2-C10 alkenyl, substituted or unsubstituted C3-C10 heterocycloalkyl, substituted or unsubstituted C3-C10 heterocycloalkenyl, substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl; R2, R4 and R5 are each independently H, halogen, -CN, substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C2-C10 alkenyl, substituted or unsubstituted C3-C10 heterocycloalkyl, substituted or unsubstituted C3-C10 heterocycloalkenyl, substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl. R3 is H, D, halogen, -CN, substituted or unsubstituted C1-C10 alkyl, -CD3, substituted or unsubstituted C2-C10 alkynyl, substituted or unsubstituted C3-C10 cycloalkyl, substituted or unsubstituted C3-C10 cycloalkenyl, substituted or unsubstituted C3-C10 heterocycloalkyl, substituted or unsubstituted C3-C10 heterocycloalkenyl, substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl; R3 is H, D, halogen, -CN, substituted or unsubstituted C1-C10 alkyl, -CD3, substituted or unsubstituted C2-C10 alkene R6 is H or D; and m and n are each independently 0, 1, 2 or 3, provided that at least one of m and n is not zero.

[0011] The present disclosure also relates to methods for treating a disease in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein, such as a compound of formula (1) disclosed herein, or a pharmaceutical composition thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 Schematic diagram of Formula 1. The wavy line represents a group attached to the ring structure in any stereochemical configuration. The bond between positions 5 and 6 can be a single bond (CC) or a double bond (C=C).

[0013] Figure 2A-2F .Representative examples of compounds having R3 groups.

[0014] Figure 3A-3F . Additional representative examples of compounds wherein R3 is -CH3.

[0015] Figures 4A-4F . Additional representative examples of compounds wherein R3 is cycloalkyl.

[0016] Figures 5A-5L . Representative examples of compounds having RX group or halogen X.

[0017] Figures 6A-6F . Representative examples of compounds wherein the RX group is -CFH2.

[0018] Figures 7A-7L . Representative examples of compounds having a heterocycle as R3.

[0019] Figures 8A-8F . Representative examples of compounds having a specific heterocycle as R3. DETAILED DESCRIPTION

[0020] The following is a more detailed description of various concepts related to the method and apparatus according to the present disclosure and embodiments of the method and apparatus according to the present disclosure. It should be understood that the various aspects of the subject matter introduced above and discussed in more detail below can be implemented in any of many ways, as the subject matter is not limited to any particular embodiment. Examples of specific embodiments and applications are provided primarily for illustrative purposes.

[0021] As used herein, the term "gamma-aminobutyric acid type A receptor," "GABA A receptors", "GABA A Rs", "GABA A R", "GABAARs", "GABAARs" or their grammatical variations, whether singular or plural, refer to gamma-aminobutyric acid type A receptors (GABA A R), which is a type of receptor that responds to the neurotransmitter gamma-aminobutyric acid (GABA). GABA is the main inhibitory neurotransmitter in the cerebral cortex, which is important for maintaining an inhibitory state that balances neuronal excitation. GABA A Disturbances in receptors or an imbalance in GABA and neural excitation can lead to widespread disturbances in brain circuits and GABA-related functions, which are important for various behavioral states such as anxiety levels, panic, stress responses, seizures, sleep, vigilance, and memory.

[0022] Many natural and synthetic neuroactive steroids can interact with GABA A R binds to and regulates its activity.

[0023] As used herein, the term "neuroactive steroid," "NAS," "neuroactive steroids," "NASs," or variations thereof, refers to a steroid that inhibits neurotransmission, particularly GABA. AOne or more neurosteroids (NS) that inhibit agonists of the agonist receptors. In some embodiments, neuroactive steroids act as modulators of the gamma-aminobutyric acid (GABA) receptor complex (GRC) in the central nervous system (CNS). Examples include, but are not limited to, tetrahydrodeoxycorticosterone (THDOC), androstane, androstane 3α-androstanediol, cholestane cholesterol, pregnane, pregnane pregnanolone (eltanolone), allopregnanolone, brexanolone, ganaxolone, and SAGE-217.

[0024] As used herein, the term "alkyl" refers to a monovalent straight or branched saturated aliphatic carbon chain or group. For example, "C1-C10 alkyl" (or "C1-C10 alkyl" 10 "alkyl") refers to any alkyl group having 1 to 10 carbon atoms, which is a straight or branched chain, such as -CH3, -C2H5, -C3H7, -C4H9, -C5H 11 、-C6H 13 、-C7H 15 、-C8H 17 、-C9H 19 or -C 10 H 21 As another example, "C1-C4 alkyl" refers to n-butyl, isobutyl, sec-butyl and tert-butyl, n-propyl and isopropyl, ethyl or methyl.

[0025] As used herein, the term "alkylene" or "alkylene chain" refers to a fully saturated straight or branched divalent hydrocarbon chain group having one to twelve carbon atoms. Non-limiting examples of C1-C10 alkylene groups include methylene, ethylene, propylene, n-butylene, etc. The alkylene chain can be connected to the rest of the molecule by a single bond and to a group (e.g., those described herein) by a single bond. The connection point of the alkylene chain to the rest of the molecule and to the group can be through one carbon or any two carbons within the chain. Unless otherwise specifically stated in the specification, the alkylene chain may be optionally substituted.

[0026] As used herein, the term "alkenyl" refers to a straight or branched aliphatic hydrocarbon group containing at least one carbon-carbon double bond and having a number of carbon atoms within the specified range. For example, "C2-C10 alkenyl" (or "C2-C10 alkenyl" 10 "Alkenyl" refers to any alkenyl group having 2 to 6 carbon atoms that is linear or branched, or an isomer. In another example, C2-C10 alkenyl may refer to 1-butenyl, 2-butenyl, 3-butenyl, isobutenyl, 1-propenyl, 2-propenyl, or ethenyl (or vinyl).

[0027] As used herein, the term "alkynyl" refers to a group of straight or branched hydrocarbon groups having at least one or more carbon-carbon triple bonds and optionally one or more carbon-carbon double bonds. In some instances, a C2-C10 alkynyl group may have 2 to 10 carbon atoms, one or more carbon-carbon triple bonds (e.g., 1, 2, 3, or 4 carbon-carbon triple bonds), and optionally one or more carbon-carbon double bonds (e.g., 1, 2, 3, or 4 carbon-carbon double bonds). In certain instances, an alkynyl group may not contain a double bond. In some instances, an alkynyl group has 2 to 10 carbon atoms ("C2-C10 alkynyl"). In yet other instances, an alkynyl group has 2 to 9 carbon atoms ("C2-C9 alkynyl"). In yet other instances, an alkynyl group has 2 to 8 carbon atoms ("C2-C8 alkynyl"). In yet other instances, an alkynyl group has 2 to 7 carbon atoms ("C2-C7 alkynyl"). In yet other examples, the alkynyl group has 2 to 6 carbon atoms ("C2-C6 alkynyl"). In yet other examples, the alkynyl group has 2 to 5 carbon atoms ("C2-C5 alkynyl"). In yet other examples, the alkynyl group has 2 to 4 carbon atoms ("C2-C4 alkynyl"). In yet other examples, the alkynyl group has 2 to 3 carbon atoms ("C2-C3 alkynyl"). In yet other examples, the alkynyl group has 2 carbon atoms ("C2 alkynyl"). The one or more carbon-carbon triple bonds can be internal (such as in 2-butynyl) or terminal (such as in 1-butynyl). Examples of C2-C4 alkynyl groups include, but are not limited to, ethynyl (C2), 1-propynyl (C3), 2-propynyl (C3), 1-butynyl (C4), 2-butynyl (C4), and the like. Examples of C2-C6 alkynyl groups can include the above-mentioned C2-C4 alkynyl groups as well as pentynyl (C5), hexynyl (C6), and the like. Additional examples of alkynyl groups can include heptynyl (C7), octynyl (C8), and the like. Unless otherwise indicated, each example of an alkynyl group can independently and optionally be substituted, i.e., unsubstituted ("unsubstituted alkynyl") or substituted ("substituted alkynyl") with one or more substituents; for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. In certain embodiments, an alkynyl group is an unsubstituted C2-C10 alkynyl group. In certain embodiments, an alkynyl group is a substituted C2-C10 alkynyl group.

[0028] As used herein, the term "cycloalkyl" refers to any monocyclic ring of an alkane having a number of carbon atoms within the specified range. For example, "C3-C10 cycloalkyl" (or "C3-C10 cycloalkyl" 10 "Cycloalkyl" refers to a monocyclic ring of an alkane having 3 to 6 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl.

[0029] As used herein, the term "heterocycloalkyl", "heterocycloalkenyl" or "heterocycle" refers to a ring structure having carbon atoms and one or more heteroatoms selected from N, O, S, boron, silicon, phosphorus or a combination thereof as ring structure members. In some embodiments, "heterocycloalkyl" is a "C3-C10 heterocycloalkyl" (or C3-C10 heterocycloalkyl) containing one or more heteroatoms such as N, O, S or a combination thereof. 10 The heterocycloalkyl group may have one or more substitutions. The substitutions may be on one or more carbon atoms or any heteroatom.

[0030] As used herein, the term "haloalkyl" refers to an alkyl group as defined above that is substituted with one or more halogen atoms (e.g., F, Cl, Br, and I). When a haloalkyl group comprises two or more halogen atoms, the halogen atoms may be the same or different. Non-limiting examples of haloalkyl groups include trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, etc. Unless otherwise specifically stated in the specification, a haloalkyl group may be optionally substituted.

[0031] As used herein, the term "heteroalkyl" refers to an alkyl group that further comprises one or more heteroatoms (e.g., N, O, S, boron, silicon, phosphorus, or a combination thereof) within the parent chain, wherein one or more heteroatoms are inserted between adjacent carbon atoms within the parent carbon chain, and / or one or more heteroatoms are inserted between the carbon atom and the parent molecule, i.e., between the points of attachment. In certain instances, heteroalkyl refers to a saturated group ("C1-C10 heteroalkyl") having 1 to 10 carbon atoms and one or more heteroatoms. In some other instances, heteroalkyl is a saturated group ("C1-C9 heteroalkyl") having 1 to 9 carbon atoms and one or more heteroatoms. In other instances, heteroalkyl is a saturated group ("C1-C8 heteroalkyl") having 1 to 8 carbon atoms and one or more heteroatoms. In yet other instances, heteroalkyl is a saturated group ("C1-C7 heteroalkyl") having 1 to 7 carbon atoms and one or more heteroatoms. In yet other examples, heteroalkyl groups are groups having 1 to 6 carbon atoms and one or more heteroatoms (“C1-C6 heteroalkyl”). In yet other examples, heteroalkyl groups are saturated groups having 1 to 5 carbon atoms and 1 or more heteroatoms (“C1-C5 heteroalkyl”). In yet other examples, heteroalkyl groups are saturated groups having 1 to 4 carbon atoms and 1 or 2 heteroatoms (“C1-C4 heteroalkyl”). In yet other examples, heteroalkyl groups are saturated groups having 1 to 3 carbon atoms and 1 heteroatom (“C1-C3 heteroalkyl”). In yet other examples, heteroalkyl groups are saturated groups having 1 to 2 carbon atoms and 1 heteroatom (“C1-C2 heteroalkyl”). In yet other examples, heteroalkyl groups are saturated groups having 1 carbon atom and 1 heteroatom (“C1 heteroalkyl”). In yet other examples, heteroalkyl groups are saturated groups having 2 to 6 carbon atoms and 1 or 2 heteroatoms (“C2-C6 heteroalkyl”). Unless otherwise specified, each instance of a heteroalkyl group is independently unsubstituted (an "unsubstituted heteroalkyl group") or substituted (a "substituted heteroalkyl group") with one or more substituents. In yet other instances, the heteroalkyl group is an unsubstituted C1-C10 heteroalkyl group. In even other instances, the heteroalkyl group is a substituted C1-C10 heteroalkyl group. The substitution may be on one or more carbon atoms or any heteroatom.

[0032] As used herein, the term "heteroalkenyl" refers to an alkenyl group that further includes one or more (e.g., 1, 2, 3, or 4) heteroatoms (e.g., N, O, S, boron, silicon, phosphorus, or a combination thereof), wherein one or more heteroatoms are inserted between adjacent carbon atoms within the parent carbon chain, and / or one or more heteroatoms are inserted between a carbon atom and the parent molecule, i.e., between the points of attachment. In some instances, heteroalkenyl refers to a group having 2 to 10 carbon atoms, at least one double bond, and 1, 2, 3, or 4 heteroatoms ("C2-C10 heteroalkenyl"). In other instances, heteroalkenyl has 2 to 9 carbon atoms, at least one double bond, and 1, 2, 3, or 4 heteroatoms ("C2-C9 heteroalkenyl"). In yet other instances, heteroalkenyl has 2 to 8 carbon atoms, at least one double bond, and 1, 2, 3, or 4 heteroatoms ("C2-C8 heteroalkenyl"). In yet other examples, heteroalkenyl groups have 2 to 7 carbon atoms, at least one double bond, and 1, 2, 3, or 4 heteroatoms (“C-C heteroalkenyl”). In yet other examples, heteroalkenyl groups have 2 to 6 carbon atoms, at least one double bond, and 1, 2, or 3 heteroatoms (“C-C heteroalkenyl”). In yet other examples, heteroalkenyl groups have 2 to 5 carbon atoms, at least one double bond, and 1 or 2 heteroatoms (“C-C heteroalkenyl”). In yet other examples, heteroalkenyl groups have 2 to 4 carbon atoms, at least one double bond, and 1 or 2 heteroatoms (“C-C heteroalkenyl”). In yet other examples, heteroalkenyl groups have 2 to 3 carbon atoms, at least one double bond, and 1 heteroatom (“C-C heteroalkenyl”). In yet other examples, heteroalkenyl groups have 2 to 6 carbon atoms, at least one double bond, and 1 or 2 heteroatoms (“C-C heteroalkenyl”). Unless otherwise specified, each instance of a heteroalkenyl group is independently unsubstituted (an "unsubstituted heteroalkenyl group") or substituted (a "substituted heteroalkenyl group") with one or more substituents. In yet other instances, a heteroalkenyl group is an unsubstituted C2-C10 heteroalkenyl group. In even other instances, a heteroalkenyl group is a substituted C2-C10 heteroalkenyl group. Substitutions may be on one or more carbon atoms or any heteroatom.

[0033] As used herein, the term "heteroalkynyl" refers to an alkynyl group that further comprises one or more heteroatoms (e.g., N, O, S, boron, silicon, phosphorus, or a combination thereof), wherein the one or more heteroatoms are inserted between adjacent carbon atoms within the parent carbon chain, and / or the one or more heteroatoms are inserted between a carbon atom and the parent molecule, i.e., between the points of attachment. In certain instances, a heteroalkynyl group refers to a group having 2 to 10 carbon atoms, at least one triple bond, and 1, 2, 3, or 4 heteroatoms ("C2-C10 heteroalkynyl"). In other instances, a heteroalkynyl group has 2 to 9 carbon atoms, at least one triple bond, and 1, 2, 3, or 4 heteroatoms ("C2-C9 heteroalkynyl"). In yet other instances, a heteroalkynyl group has 2 to 8 carbon atoms, at least one triple bond, and 1, 2, 3, or 4 heteroatoms ("C2-C8 heteroalkynyl"). In yet other examples, the heteroalkynyl group has 2 to 7 carbon atoms, at least one triple bond, and 1, 2, 3, or 4 heteroatoms (“C-C heteroalkynyl”). In yet other examples, the heteroalkynyl group has 2 to 6 carbon atoms, at least one triple bond, and 1, 2, or 3 heteroatoms (“C-C heteroalkynyl”). In yet other examples, the heteroalkynyl group has 2 to 5 carbon atoms, at least one triple bond, and 1 or 2 heteroatoms (“C-C heteroalkynyl”). In yet other examples, the heteroalkynyl group has 2 to 4 carbon atoms, at least one triple bond, and 1 or 2 heteroatoms (“C-C heteroalkynyl”). In yet other examples, the heteroalkynyl group has 2 to 3 carbon atoms, at least one triple bond, and 1 heteroatom (“C-C heteroalkynyl”). In yet other examples, the heteroalkynyl group has 2 to 6 carbon atoms, at least one triple bond, and 1 or 2 heteroatoms (“C-C heteroalkynyl”). Unless otherwise specified, each instance of a heteroalkynyl group is independently unsubstituted (an "unsubstituted heteroalkynyl group") or substituted (a "substituted heteroalkynyl group") with one or more substituents. In yet other instances, the heteroalkynyl group is an unsubstituted C2-C10 heteroalkynyl group. In even other instances, the heteroalkynyl group is a substituted C2-C10 heteroalkynyl group. The substitution may be on one or more carbon atoms or any heteroatom.

[0034] As used herein, the term "cycloalkylalkyl" refers to an alkyl group in which the alkyl group is substituted by a cycloalkyl group. Typical cycloalkylalkyl groups include, but are not limited to, cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, cycloheptylmethyl, cyclooctylmethyl, cyclopropylethyl, cyclobutylethyl, cyclopentylethyl, cyclohexylethyl, cycloheptylethyl, and cyclooctylethyl. Cycloalkylalkyl groups may be unsubstituted or substituted. Substitutions may be on one or more carbon atoms or any heteroatom.

[0035] As used herein, the term "heterocyclylalkyl" refers to an alkyl group in which the alkyl group is substituted with a heterocyclyl group. Typical heterocyclylalkyl groups include, but are not limited to, pyrrolidinylmethyl, piperidinylmethyl, piperazinylmethyl, morpholinylmethyl, pyrrolidinylethyl, piperidinylethyl, piperazinylethyl, morpholinylethyl, and the like. Heterocyclylalkyl groups may be unsubstituted or substituted. Substitutions may be on one or more carbon atoms or any heteroatom.

[0036] As used herein, the term "cycloalkenyl" refers to a substituted or unsubstituted carbocyclic group having 3 to 10 carbon atoms and having a single ring or multiple fused rings, including fused rings and bridged ring systems, and having at least one and particularly 1 to 2 sites of olefinic unsaturation. For example, such cycloalkenyl groups include single ring structures such as cyclohexenyl, cyclopentenyl, cyclopropenyl, etc. Cycloalkenyl groups can be unsubstituted or substituted. Substitution can be on one or more carbon atoms or any heteroatom.

[0037] As used herein, the term "aryl" refers to a cyclic or one or more fused hydrocarbon ring systems in which at least one ring is aromatic. The term "heteroaryl" refers to a heteroaromatic ring containing one or more, such as 1 to 4, heteroatoms independently selected from N, O, and S, wherein each N is optionally in the form of an oxide to the extent chemically possible. Aryl or heteroaryl groups may be substituted or unsubstituted. Substitutions may be on one or more carbon atoms or any heteroatom.

[0038] As used herein, the term "halogen" (or "halo") refers to fluorine, chlorine, bromine, and iodine (alternatively referred to as fluorine (-F), chlorine (-Cl), bromine (-Br), and iodine (-I)).

[0039] As used herein, the term "isomer" refers to structural isomers, such as groups or atoms located at different positions in a molecule; stereoisomers, such as chiral isomers, enantiomers, diastereomers, and cis / trans isomers; tautomers; or combinations thereof. Mixtures of isomers may also be suitable. An isomer mixture may contain all proportions of the corresponding isomers. Salts of isomers may also be suitable. The neuroactive steroids of the present invention may comprise an isomer thereof, one or more salts thereof, one or more solvates including hydrates thereof, solvated salts thereof, or mixtures thereof. Absolute stereochemistry or isomeric configuration may be determined by X-ray crystallography, by vibrational circular dichroism (VCD) spectroscopy analysis, or a combination thereof. Isomers that have the desired biological activity in vivo may be particularly preferred.

[0040] The neuroactive steroids disclosed herein can be identified by names based on the nomenclature recommended by the International Union of Pure and Applied Chemistry (IUPAC) or other nomenclature systems. These compounds can also be identified by chemical structure diagrams. Unless clearly stated to the contrary in a particular context, these names and structures are used interchangeably throughout this disclosure.

[0041] As used herein, "effective amount" refers to a therapeutically effective amount or a prophylactic effective amount. "Therapeutically effective amount" refers to an amount that effectively achieves the desired therapeutic outcome, such as tumor size reduction, life extension, or life expectancy extension, at the necessary dosage and time period. The therapeutically effective amount of a compound can vary according to factors such as disease state, age, sex, and subject weight, as well as the ability of the compound to elicit the desired response in the subject. The dosage regimen can be adjusted to provide the best therapeutic response. A therapeutically effective amount is also the amount in which the therapeutically beneficial effects of the compound exceed any toxic or deleterious effects thereof. A "prophylactic effective amount" refers to an amount that effectively achieves the desired prophylactic outcome (such as tumor shrinkage, life extension, life expectancy extension, or prevention of prostate cancer progression to a castration-resistant form) at the necessary dosage and time period. Typically, a prophylactic dose is used for a subject before the disease or in the early stages of the disease so that the prophylactic effective amount can be less than the therapeutically effective amount.

[0042] As used herein, "treating" or "treatment" encompasses the treatment of a disease or condition of interest in a mammal, such as a human, suffering from the disease or condition of interest, and includes, but is not limited to:

[0043] 1. Preventing the disease or condition from occurring in a mammal, particularly when such mammal is susceptible to the condition but has not yet been diagnosed as having the condition;

[0044] 2. To inhibit the disease or condition, i.e. to prevent its development;

[0045] 3. Alleviate the disease or condition, i.e., cause regression of the disease or condition (ranging from reducing the severity of the disease or condition to curing the disease or condition); or

[0046] 4. Relieve symptoms caused by the disease or condition, i.e., relieve pain without addressing the underlying disease or condition.

[0047] As used herein, the terms "disease" and "condition" may be used interchangeably or may be distinct in that a particular disease or condition has no known etiology (such that an etiology has not yet been determined) and therefore has not been recognized as a disease, but only as an undesirable condition or syndrome in which clinicians have identified a more or less specific group of symptoms.

[0048] As used herein, a "subject" can be a human, non-human primate, mammal, rat, mouse, cow, horse, pig, sheep, goat, dog, cat, insect, etc. The subject may be suspected of having cancer (such as leukemia) or another disease or condition or be at risk of having cancer (such as leukemia) or another disease or condition. Methods for diagnosing various cancers and clinical descriptions of cancer are known to those of ordinary skill in the art. The subject may also be suspected of having an infection or cardiovascular dysfunction.

[0049] Unless otherwise expressly stated, all ranges cited herein are inclusive. For example, a heterocycle described as being included in the range of "1 to 4 heteroatoms" means that the ring may contain 1, 2, 3 or 4 heteroatoms. It should also be understood that any range cited herein includes within its scope all subranges within that range. Thus, for example, a heterocycle described as containing "1 to 4 heteroatoms" is intended to include heterocycles containing 2 to 4 heteroatoms, 3 or 4 heteroatoms, 1 to 3 heteroatoms, 2 or 3 heteroatoms, 1 or 2 heteroatoms, 1 heteroatom, 2 heteroatoms, 3 heteroatoms or 4 heteroatoms as aspects thereof. In other examples, C1-C10 alkyl means that the alkyl group contains 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10 carbon atoms, including all subranges. Thus, C1-C10 alkyl can be methyl, ethyl, propyl, C4 alkyl, C5 alkyl, C6 alkyl, C7 alkyl, C8 alkyl, C9 alkyl and C10 alkyl. In addition, each C1-C10 alkyl group can independently be straight chain or branched. Similarly, C2-C10 alkenyl means a straight chain or branched alkenyl group containing 2, 3, 4, 5, 6, 7, 8, 9 and 10 carbon atoms. Straight chain or branched alkenyl groups can be suitable. C3-C10 cycloalkyl means a straight chain or branched cycloalkyl group containing 3, 4, 5, 6, 7, 8, 9 and 10 carbon atoms.

[0050] The present disclosure relates to neuroactive steroids (NAS) of formula (1):

[0051]

[0052] one or more isomers thereof, deuterium-labeled variants thereof, or pharmaceutically acceptable salts thereof,

[0053] in:

[0054] R1 is H, D, substituted or unsubstituted C1-C10 alkyl, C1-C5 deuterated alkyl, substituted or unsubstituted C2-C10 alkenyl, substituted or unsubstituted C2-C10 alkynyl, substituted or unsubstituted C3-C10 cycloalkyl, substituted or unsubstituted C3-C10 cycloalkenyl, substituted or unsubstituted C3-C10 heterocycloalkyl, substituted or unsubstituted C3-C10 heterocycloalkenyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;

[0055] R2, R4 and R5 are each independently H, halogen, -CN, substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C2-C10 alkenyl, substituted or unsubstituted C2-C10 alkynyl, substituted or unsubstituted C3-C10 cycloalkyl, substituted or unsubstituted C3-C10 cycloalkenyl, substituted or unsubstituted C3-C10 heterocycloalkyl, substituted or unsubstituted C3-C10 heterocycloalkenyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl;

[0056] R3 is H, D, halogen, -CN, substituted or unsubstituted C1-C10 alkyl, -CD3, substituted or unsubstituted C2-C10 alkenyl, substituted or unsubstituted C2-C10 alkynyl, substituted or unsubstituted C3-C10 cycloalkyl, substituted or unsubstituted C3-C10 cycloalkenyl, substituted or unsubstituted C3-C10 heterocycloalkyl, substituted or unsubstituted C3-C10 heterocycloalkenyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;

[0057] R6 is H or D; and

[0058] m and n are each independently an integer 0, 1, 2 or 3, provided that at least one of m and n is non-zero.

[0059] Any atom in the compounds disclosed herein may exhibit their natural isotopic abundance, or one or more atoms may be artificially enriched with a particular isotope having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number predominantly found in nature. The present invention is intended to include all suitable isotopic variations of the compounds disclosed herein.

[0060] In some embodiments, the compounds of the present disclosure include all isotopes of atoms present in the intermediates or final compounds. Isotopes include those atoms with the same atomic number but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium. In some embodiments, compounds disclosed herein, such as compounds of formula (1), include one or more deuterium atoms. In some embodiments, one or more of R1, R2, R3, R4 and R5 are deuterium atoms or deuterated alkyl groups (e.g., C1-C10 deuterated alkyl groups or C1-C5 deuterated alkyl groups). In some embodiments, one or more of R1, R2, R3, R4 and R5 are deuterium atoms or -CD3. In some embodiments of formula (1), R3 is a deuterium atom or a deuterated alkyl group, and one or more CH bonds of formula (1) are replaced by CD bonds.

[0061] Figure 1 A schematic diagram of a compound of formula (1) is shown in FIG. 1 , wherein the positions of carbon atoms are indicated by numbers. If a chiral center is present at that position, then Figure 1The wavy line in the figure represents a group attached to the ring structure at the chiral center. The bond between positions 5 and 6, as represented by a pair of solid and dashed lines, can be a single bond (C-C) or a double bond (C=C). In some embodiments, the bond between positions 5 and 6 is a C-C bond. In some embodiments, the bond between positions 5 and 6 is a C=C double bond.

[0062] In some embodiments of formula (1), R1 is H, D, substituted or unsubstituted C1-C10 alkyl, -CD3, substituted or unsubstituted C2-C10 alkenyl, substituted or unsubstituted C2-C10 alkynyl, substituted or unsubstituted C3-C10 cycloalkyl, substituted or unsubstituted C3-C10 cycloalkenyl, substituted or unsubstituted C3-C10 heterocycloalkyl, substituted or unsubstituted C3-C10 heterocycloalkenyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl. In some embodiments, R1 is H, substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C2-C10 alkenyl, substituted or unsubstituted C2-C10 alkynyl, substituted or unsubstituted C3-C10 cycloalkyl, substituted or unsubstituted C3-C10 cycloalkenyl, substituted or unsubstituted C3-C10 heterocycloalkyl, substituted or unsubstituted C3-C10 heterocycloalkenyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl. In some embodiments, R1 is H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, or substituted or unsubstituted cycloalkyl. In some embodiments, R1 is independently H, D, substituted or unsubstituted alkyl, -CD3, or substituted or unsubstituted cycloalkyl. In some embodiments, R1 is independently H, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl. In some embodiments, R1 is independently H, D, substituted or unsubstituted alkyl, or -CD3. In some embodiments, R1 is independently H or substituted or unsubstituted alkyl. In some embodiments, R1 is independently H or unsubstituted alkyl. In some embodiments, R1 is H. In some embodiments, R1 is substituted or unsubstituted alkyl. In some embodiments, R1 is unsubstituted alkyl. In some embodiments, the alkyl group is C1-C10 alkyl. In some embodiments, the alkyl group is C1-C5 alkyl. In some embodiments, the alkyl group is methyl, ethyl, or isopropyl. In some embodiments, the alkyl group is methyl or ethyl. In some embodiments, the alkyl group is methyl. In some embodiments, the alkenyl group is C2-C10 alkenyl. In some embodiments, the alkenyl group is C2-C5 alkenyl. In some embodiments, the alkynyl group is C2-C10 alkynyl. In some embodiments, the alkynyl group is C2-C5 alkynyl. In some embodiments, the cycloalkyl group is C3-C10 cycloalkyl. In some embodiments, the cycloalkyl group is C3-C6 cycloalkyl. In some embodiments, the cycloalkyl group is cyclopropyl or cyclobutyl. In some embodiments, the cycloalkyl group is cyclopropyl.

[0063] In some embodiments of formula (1), R2, R4 and R5 are each independently H, halogen, -CN, substituted or unsubstituted C1-C5 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C3-C6 cycloalkenyl, substituted or unsubstituted C3-C6 heterocycloalkyl, substituted or unsubstituted C3-C6 heterocycloalkenyl, substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl. In some embodiments, R2, R4 and R5 are each independently H, halogen, -CN, substituted or unsubstituted C1-C5 alkyl or C3-C6 cycloalkyl. In some embodiments, R2, R4 and R5 are each independently H, halogen, -CN or substituted or unsubstituted C1-C5 alkyl. In some embodiments, R2, R4, and R5 are each independently H, halogen, or substituted or unsubstituted C1-C5 alkyl. In some embodiments, C1-C5 alkyl is methyl or ethyl. In some embodiments, C1-C5 alkyl is methyl. In some embodiments, C2-C6 alkenyl is ethenyl, propenyl, or isopropenyl. In some embodiments, substituted or unsubstituted C2-C6 alkynyl is substituted or unsubstituted ethynyl, propynyl, or butynyl. In some embodiments, substituted or unsubstituted C3-C6 cycloalkyl is substituted or unsubstituted cyclopropyl or cyclobutyl. In some embodiments, substituted or unsubstituted C3-C6 cycloalkyl is substituted or unsubstituted cyclopropyl. In some embodiments, substituted or unsubstituted C3-C6 heterocycloalkyl is substituted or unsubstituted azetidinyl, pyrrolidinyl, piperidinyl, morpholinyl, or thiomorpholinyl. In some embodiments, substituted or unsubstituted aryl is substituted or unsubstituted phenyl. In some embodiments, the substituted or unsubstituted heteroaryl is a substituted or unsubstituted oxazolyl, thiazolyl, imidazolyl, triazolyl, pyrazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, pyridinyl or pyrimidinyl. In some embodiments, R2, R4 and R5 are each independently H, halogen, -CN, -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)2, -CH=CH2, -C(Me)=CH2, -CH=CH(Me), -C≡CH, -C≡C(Me), cyclopropyl, cyclobutyl, phenyl or pyridinyl.

[0064] In some embodiments of formula (1), R3 is H, D, halogen, -CN, substituted or unsubstituted C1-C5 alkyl, deuterated C1-C5 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C3-C6 cycloalkenyl, substituted or unsubstituted C3-C6 heterocycloalkyl, substituted or unsubstituted C3-C6 heterocycloalkenyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl. In some embodiments, R3 is H, D, halogen, -CN, substituted or unsubstituted C1-C5 alkyl, -CD3, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C3-C6 cycloalkenyl, substituted or unsubstituted C3-C6 heterocycloalkyl, substituted or unsubstituted C3-C6 heterocycloalkenyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl. In some embodiments, R3 is H, halogen, -CN, substituted or unsubstituted C1-C5 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C3-C6 cycloalkenyl, substituted or unsubstituted C3-C6 heterocycloalkyl, substituted or unsubstituted C3-C6 heterocycloalkenyl, substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl. In some embodiments, R3 is H, D, halogen, -CN, substituted or unsubstituted C1-C5 alkyl, deuterated C1-C5 alkyl or C3-C6 cycloalkyl. In some embodiments, R3 is H, D, halogen, -CN, substituted or unsubstituted C1-C5 alkyl, -CD3 or C3-C6 cycloalkyl. In some embodiments, R3 is H, halogen, -CN, substituted or unsubstituted C1-C5 alkyl, or C3-C6 cycloalkyl. In some embodiments, R3 is H, D, halogen, -CN, substituted or unsubstituted C1-C5 alkyl, -CD3. In some embodiments, R3 is H, halogen, -CN, or substituted or unsubstituted C1-C5 alkyl. In some embodiments, R3 is independently H, halogen, or substituted or unsubstituted C1-C5 alkyl. In some embodiments, C1-C5 alkyl is methyl or ethyl. In some embodiments, C1-C5 alkyl is methyl. In some embodiments, halogen is F. In some embodiments, C2-C6 alkenyl is ethenyl, propenyl, or isopropenyl. In some embodiments, substituted or unsubstituted C2-C6 alkynyl is substituted or unsubstituted ethynyl, propynyl, or butynyl. In some embodiments, substituted or unsubstituted C3-C6 cycloalkyl is substituted or unsubstituted cyclopropyl or cyclobutyl. In some embodiments, the substituted or unsubstituted C3-C6 cycloalkyl is substituted or unsubstituted cyclopropyl.In some embodiments, substituted or unsubstituted C3-C6 heterocycloalkyl is substituted or unsubstituted azetidinyl, pyrrolidinyl, piperidinyl, morpholinyl or thiomorpholinyl. In some embodiments, substituted or unsubstituted aryl is substituted or unsubstituted phenyl. In some embodiments, substituted or unsubstituted heteroaryl is substituted or unsubstituted oxazolyl, thiazolyl, imidazolyl, triazolyl, pyrazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, pyridinyl or pyrimidinyl. In some embodiments, R3 is H, halogen, -CN, -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)2, -CH=CH2, -C(Me)=CH2, -CH=CH(Me), -C≡CH, -C≡C(Me), cyclopropyl, cyclobutyl, phenyl or pyridinyl.

[0065] In some embodiments of formula (1), R3 is H, halogen, substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C2-C10 alkenyl, substituted or unsubstituted C2-C10 alkynyl, substituted or unsubstituted C3-C10 cycloalkyl, substituted or unsubstituted C3-C10 cycloalkenyl, substituted or unsubstituted C3-C10 heterocycloalkyl, substituted or unsubstituted C3-C10 heterocycloalkenyl, substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl. Any of the above-mentioned substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C2-C10 alkenyl, substituted or unsubstituted C2-C10 alkynyl, substituted or unsubstituted C3-C10 cycloalkyl and substituted or unsubstituted C3-C10 cycloalkenyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl may be suitable.

[0066] In some embodiments, R3 is H, -D, -CH3, -CD3, -CN, substituted or unsubstituted cyclopropyl, substituted or unsubstituted C1-C10 haloalkyl, substituted or unsubstituted C3-C10 heterocycloalkyl, substituted or unsubstituted C3-C10 heterocycloalkenyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -X, or

[0067]

[0068] wherein X is selected from the group consisting of Cl, F, Br and I.

[0069] In some embodiments, R3 is CH3 (e.g., as in Figure 3A – Figure 3F In the formulas (8) to (13)), substituted or unsubstituted cyclopropyl (for example, as in Figure 4A – Figure 4F substituted or unsubstituted C3-C10 heterocycloalkyl, substituted or unsubstituted C3-C10 heterocycloalkenyl (e.g., Figure 7A – Figure 7L (32)-(37) and (32')-(37')), halogen (e.g., as in Figure 5G – Figure 5L In the formula (20')-(25'), wherein halogen is represented by X) or a substituted or unsubstituted C1-C10 haloalkyl group (for example, as in Figure 5A – Figure 5F In formulas (20)-(25), wherein C1-C10 halogen is represented by R). In some embodiments, halogen is Cl, F, Br, or I. In some embodiments, halogen is Cl or F. In some embodiments, halogen is Br or I. In some embodiments, halogen is Cl. In some embodiments, halogen is F. In some embodiments, halogen is Br. In some embodiments, halogen is I. In some embodiments, R3 is C1-C10 haloalkyl. In some embodiments, C1-C10 haloalkyl is -CXH2, -CX2H, -CX3, -CH2CXH2, -CH2CX2H, or -CH2CX3, wherein X is Cl, F, Br, or I. In some embodiments, C1-C10 haloalkyl is -CC1H2, -CC12H, -CC13, -CFH2, -CF2H, -CF3, -CBrH2, -CBr2H, -CBr3, -CIH2, -CI2H, -CI3, -CC1FH, -CC1BrH, -CC1(I)H, -CFBrH, -CF(I)H, -CBr(I)H, -CC12F, -CC1F2, -CC12Br, -CC1Br2, -CC12(I), -CC1(I)2, -CF2Br, -CFBr2, -CF2(I), -CF(I)2, etc. (for illustrative purposes, some iodine is shown as (I)). In some embodiments, C1-C10 haloalkyl is -CFH2 (e.g., Figures 6A-6F Formula (26)-(31)), -CF2H, -CF3, CH2CFH2, -CH2CF2H or -CH2CF3.

[0070] In some embodiments, R3 is:

[0071]

[0072] In some embodiments, R3 is selected from the group consisting of H, D, F, -CH3, -CD3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl, -CN,

[0073] In some embodiments, R3 is selected from the group consisting of H, F, -CH3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl, -CN,

[0074] In some embodiments, R3 is selected from the group consisting of H, D, F, -CH3, -CD3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl, and -CN. In some embodiments, R3 is H, D, F, -CD3, or -CN.

[0075] In some embodiments, R3 is a substituted or unsubstituted heterocycle of formula (44)-(49):

[0076]

[0077] wherein R6 is H, substituted or unsubstituted alkyl or heteroalkyl, substituted or unsubstituted alkenyl or heteroalkenyl, substituted or unsubstituted alkynyl or heteroalkynyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted heterocycloalkenyl, or a combination thereof.

[0078] In some embodiments, R3 is

[0079]

[0080] wherein p is an integer from 1 to 5. In some embodiments, p is 1. In some embodiments, p is 2. In some embodiments, p is 3. In some embodiments, p is 4. In some embodiments, p is 5.

[0081] In some embodiments, R3 is

[0082] In some embodiments, R6 is H. In some embodiments, R6 is D.

[0083] In some embodiments of formula (I), m is 0 and n is 1, 2, or 3. In some embodiments, m is 1 and n is 0, 1, 2, or 3. In some embodiments, m is 2 and n is 0, 1, 2, or 3. In some embodiments, m is 3 and n is 0, 1, 2, or 3. In some embodiments, n is 0 and m is 1, 2, or 3. In some embodiments, n is 1 and m is 0, 1, 2, or 3. In some embodiments, n is 2 and m is 0, 1, 2, or 3. In some embodiments, n is 3 and m is 0, 1, 2, or 3. In other embodiments of formula (I), m is 1 and n is 1 (e.g., as in formulas (2)-(43), (20')-(25'), and (32')-(37'), Figures 2A-8F ). In some embodiments, m is 2, and n is 1. In some embodiments, m is 3, and n is 0.

[0084] In some embodiments of formula (1), R1, R2, R4 and R5 are each selected from the group consisting of H, D, -CH3, -CD3, -C2H5, -C3H7, -C4H9, -C5H 11 and -C6H 13 ; and R3 is selected from the group consisting of H, D, F, -CH3, -CD3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl, -CN, In some embodiments, R3 is selected from the group consisting of H, D, F, -CH3, -CD3, and -CN.

[0085] In some embodiments of formula (1), R1, R2, R4 and R5 are each selected from the group consisting of H, D, -CH3, -CD3, -C2H5, -C3H7, -C4H9, -C5H 11 and -C6H 13 ; and R3 is selected from the group consisting of H, F, -CH3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl, -CN,

[0086] In some embodiments of formula (1), R1, R2, R4 and R5 are each selected from the group consisting of: H, -CH3, -C2H5, -C3H7, -C4H9, -C5H 11 and -C6H 13; and R3 is selected from the group consisting of H, F, -CH3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl, -CN,

[0087] In some embodiments of formula (1), R1, R2, R4 and R5 are each selected from the group consisting of: H, -CH3, -C2H5, -C3H7, -C4H9, -C5H 11 and -C6H 13 ; and R3 is selected from the group consisting of H, D, F, -CH3, -CD3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl and -CN.

[0088] In some embodiments of formula (1), R1 and R2 are each independently selected from the group consisting of: H, -CH3, -C2H5, -C3H7, -C4H9, -C5H 11 and -C6H 13 ; R3 is selected from the group consisting of: H, F, -CH3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl, -CN, and R4 and R5 are each H.

[0089] In some embodiments of formula (1), R1, R2, R4 and R5 are each independently H or -CH3; R3 is selected from the group consisting of H, F, -CH3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl, -CN,

[0090] In some embodiments of formula (1), R1 and R2 are each independently H or -CH3; R3 is selected from the group consisting of H, F, -CH3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl, -CN, and R4 and R5 are H.

[0091] In some embodiments of formula (1), R1, R2, R4 and R5 are each H; and R3 is selected from the group consisting of H, F, -CH3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl, -CN,

[0092] In some embodiments of formula (1), R1, R2, R4 and R5 are each selected from the group consisting of: H, -CH3, -C2H5, -C3H7, -C4H9, -C5H 11 and -C6H 13 ; and R3 is selected from the group consisting of: H, F and -CN.

[0093] In some embodiments of formula (1), R1 and R2 are each independently selected from the group consisting of: H, -CH3, -C2H5, -C3H7, -C4H9, -C5H 11 and -C6H 13 ; R3 is selected from the group consisting of: H, F and -CN; and R4 and R5 are each H.

[0094] In some embodiments of formula (1), R1, R2, R4 and R5 are each independently H or -CH3; R3 is selected from the group consisting of: H, F and -CN.

[0095] In some embodiments of formula (1), R1 and R2 are each independently H or -CH3; R3 is selected from the group consisting of: H, F, and -CN; and R4 and R5 are H.

[0096] In some embodiments of formula (1), R1, R2, R4, and R5 are each H; and R3 is selected from the group consisting of: H, F, and -CN.

[0097] In some embodiments of formula (1), m is 1, n is 1; R1, R2, R4 and R5 are each selected from the group consisting of: H, D, -CH3, -CD3, -C2H5, -C3H7, -C4H9, -C5H 11 and -C6H 13 ; and R3 is selected from the group consisting of H, D, F, -CH3, -CD3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl, -CN,

[0098] In some embodiments of formula (1), m is 1, n is 1; R1, R2, R4 and R5 are each selected from the group consisting of: H, -CH3, -C2H5, -C3H7, -C4H9, -C5H 11 and -C6H 13 ; and R3 is selected from the group consisting of H, D, F, -CH3, -CD3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl, -CN,

[0099] In some embodiments of formula (1), m is 1, n is 1; R1, R2, R4 and R5 are each selected from the group consisting of: H, -CH3, -C2H5, -C3H7, -C4H9, -C5H 11 and -C6H 13 ; and R3 is selected from the group consisting of H, F, -CH3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl, -CN,

[0100] In some embodiments of formula (1), m is 1, n is 1; R1, R2, R4 and R5 are each selected from the group consisting of: H, D, -CH3, -CD3, -C2H5, -C3H7, -C4H9, -C5H 11 and -C6H 13 ; and R3 is selected from the group consisting of H, D, F, -CH3, -CD3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl and -CN.

[0101] In some embodiments of formula (1), m is 1, n is 1; R1, R2, R4 and R5 are each selected from the group consisting of: H, D, -CH3, -CD3, -C2H5, -C3H7, -C4H9, -C5H 11 and -C6H 13 ; and R3 is selected from the group consisting of H, F, -CH3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl and -CN.

[0102] In some embodiments of formula (1), m is 1, n is 1; R1, R2, R4 and R5 are each selected from the group consisting of H and -CH3; and R3 is selected from the group consisting of H, F, -CH3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl, -CN,

[0103] In some embodiments of formula (1), m is 1, n is 1; R1, R2, R4 and R5 are each H; and R3 is selected from the group consisting of H, F, -CH3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl, -CN,

[0104] In some embodiments of formula (1), m is 1, n is 1; R1, R2, R4, and R5 are each selected from the group consisting of H and -CH3; and R3 is selected from the group consisting of H, F, and -CN.

[0105] In some embodiments of formula (1), m is 2, n is 1; R1, R2, R4 and R5 are each selected from the group consisting of: H, -CH3, -C2H5, -C3H7, -C4H9, -C5H 11 and -C6H 13 ; and R3 is selected from the group consisting of H, F, -CH3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl, -CN,

[0106] In some embodiments of formula (1), m is 2, n is 1; R1, R2, R4 and R5 are each selected from the group consisting of H and -CH3; and R3 is selected from the group consisting of H, F, -CH3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl, -CN,

[0107] In some embodiments of formula (1), m is 2, n is 1; R1, R2, R4 and R5 are each H; and R3 is selected from the group consisting of H, F, -CH3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl, -CN,

[0108] In some embodiments of formula (1), m is 3, n is 0; R1, R2, R4 and R5 are each selected from the group consisting of: H, -CH3, -C2H5, -C3H7, -C4H9, -C5H 11 and -C6H 13 ; and R3 is selected from the group consisting of H, F, -CH3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl, -CN,

[0109] In some embodiments of formula (1), m is 3, n is 0; R1, R2, R4 and R5 are each selected from the group consisting of H and -CH3; and R3 is selected from the group consisting of H, F, -CH3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl, -CN,

[0110] In some embodiments of formula (1), m is 3, n is 0; R1, R2, R4 and R5 are each H; and R3 is selected from the group consisting of H, F, -CH3, -CH2-cyclopropyl, -CH2OH, -COOH, -CH2CN, -CH2F, -CHF2, -CH2CF3, -C≡CH, -cyclopropyl, -CN,

[0111] In some embodiments, the neuroactive steroid of formula (1) has a structure according to:

[0112] one or more isomers thereof or pharmaceutically acceptable salts thereof.

[0113] In some embodiments, the neuroactive steroid of formula (1) has a structure according to:

[0114] one or more isomers thereof or pharmaceutically acceptable salts thereof.

[0115] Neuroactive steroids suitable for use in the present invention may include at least one of R1, R2, R3, R4, and R5, which is a substituted or unsubstituted C3-C10 cycloalkyl group, a substituted or unsubstituted C3-C10 cycloalkenyl group, a substituted or unsubstituted C3-C10 heterocycloalkyl group, a substituted or unsubstituted C3-C10 heterocycloalkenyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted heteroaryl group, or a combination thereof. At least one of R1, R2, R3, R4, and R5 may be a C1-C10 haloalkyl group, wherein the halogen is selected from the group consisting of Cl, F, Br, and I. The halogen substitution may be on one or more carbon atoms. A single carbon atom may have one or more halogen substitutions, which may be the same or different.

[0116] In some embodiments, the neuroactive steroids of the present disclosure have a structure according to:

[0117]

[0118]

[0119] or a pharmaceutically acceptable salt thereof, wherein R3, m and n are as defined above in formula (1).

[0120] In some embodiments of Formula (1A)-Formula (1F), m is 0 and n is 1, 2, or 3. In some embodiments, m is 1 and n is 0, 1, 2, or 3. In some embodiments, m is 2 and n is 0, 1, 2, or 3. In some embodiments, m is 3 and n is 0, 1, 2, or 3. In some embodiments, n is 0 and m is 1, 2, or 3. In some embodiments, n is 1 and m is 0, 1, 2, or 3. In some embodiments, n is 2 and m is 0, 1, 2, or 3. In some embodiments, n is 3 and m is 0, 1, 2, or 3. In some embodiments, m is 1 and n is 1. In some embodiments, m is 2 and n is 1. In some embodiments, m is 3 and n is 0.

[0121] In some embodiments, the neuroactive steroids of the present disclosure have a structure according to:

[0122]

[0123]

[0124] or a pharmaceutically acceptable salt thereof, wherein R3 is as defined above in formula (1).

[0125] In some embodiments, the neuroactive steroids disclosed herein are compounds of formula (38)-(43), such as Figures 8A-8F shown.

[0126] In some embodiments, the neuroactive steroids disclosed herein are compounds of formula (2)-(43), (20')-(25'), and (32')-(37'), such as Figures 2A-8F shown.

[0127] In some embodiments, the neuroactive steroid of formula (1) is a compound of Table 1 shown below, or a pharmaceutically acceptable salt thereof.

[0128] Table 1. Compounds of the present disclosure

[0129]

[0130]

[0131]

[0132]

[0133] The present disclosure also relates to pharmaceutical compositions for treating diseases.

[0134] In some embodiments, a pharmaceutical composition comprises a neuroactive steroid (NAS) disclosed herein, one or more isomers thereof, a pharmaceutically acceptable salt thereof, or a combination thereof; and a pharmaceutically acceptable excipient.

[0135] In some embodiments, the pharmaceutical composition comprises a compound of formula (1), one or more isomers thereof, a pharmaceutically acceptable salt thereof, or a combination thereof; and a pharmaceutically acceptable excipient.

[0136] In some embodiments, the pharmaceutical compositions of the present disclosure comprise a compound of formula (1), wherein at least one of R1, R2, R3, R4, and R5 is a substituted or unsubstituted C3-C10 cycloalkyl, a substituted or unsubstituted C3-C10 cycloalkenyl, a substituted or unsubstituted C3-C10 heterocycloalkyl, a substituted or unsubstituted C3-C10 heterocycloalkenyl, a substituted or unsubstituted aryl, or a substituted or unsubstituted heteroaryl.

[0137] In some embodiments, the pharmaceutical compositions of the present disclosure comprise a compound of formula (1), wherein at least one of R1, R2, R3, R4, and R5 is a C1-C10 haloalkyl group, wherein the halogen is one or more Cl, F, Br, I, or a combination thereof. The halogen substitution may be on one or more carbon atoms. In some embodiments, a carbon atom has one or more halogen substitutions that are the same or different.

[0138] In some embodiments, the pharmaceutical compositions of the present disclosure comprise a compound of Formula (1A), Formula (1B), Formula (1C), Formula (1D), Formula (1E), or Formula (1F), or a pharmaceutically acceptable salt thereof, or a combination thereof; and a pharmaceutically acceptable excipient.

[0139] In some embodiments, the pharmaceutical compositions of the present disclosure comprise a compound of Formula (2) to Formula (7) or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable excipient.

[0140] In some embodiments, the pharmaceutical compositions of the present disclosure comprise compounds of formula (38)-(43), such as Figures 8A-8F As shown; and a pharmaceutically acceptable excipient.

[0141] In some embodiments, the pharmaceutical compositions of the present disclosure comprise compounds of formula (2)-(43), (20')-(25') and (32')-(37'), such as Figures 2A-8F As shown; and a pharmaceutically acceptable excipient.

[0142] In some embodiments, a pharmaceutical composition of the present disclosure comprises a compound of Table 1; and a pharmaceutically acceptable excipient.

[0143] The pharmaceutical compositions of the present disclosure may also comprise any combination of the above-described neuroactive steroids, such as those shown in formulas (1), (2)-(43), (20')-(25'), and (32')-(37'), and any of the other formulae disclosed herein.

[0144] In some embodiments, the pharmaceutical composition comprises a NAS compound disclosed herein, two or more NAS compounds disclosed herein, three or more NAS compounds disclosed herein, or four or more NAS compounds disclosed herein.

[0145] In some embodiments, pharmaceutically acceptable excipients include surfactants, emulsifiers, fillers, carriers, isotonicity agents, dispersants, viscosity modifiers, resuspending agents, buffers, or combinations thereof.

[0146] Pharmaceutical excipients generally do not have the properties of a medicine or pharmaceutically active ingredient (also referred to as an active pharmaceutical ingredient (API)), and are generally used to simplify the manufacturing process or packaging of the active ingredient, or for delivering the API to a patient or other subject. Pharmaceutically acceptable carriers, excipients, or inactive ingredients from the inactive ingredient database (https: / / www.fda.gov / drugs / drug-approvals-and-databases / inactive-ingredients-database-download) available from the U.S. FDA may be suitable. Some food substances that are generally recognized as safe (GRAS) obtained from the GRAS substance (SCOGS) database (https: / / www.fda.gov / food / generally-recognized-safe-gras / gras-substances-scogs-database) of the U.S. FDA may also be suitable.

[0147] In some embodiments, the pharmaceutically acceptable excipient is a pharmaceutically acceptable carrier. In embodiments of the present disclosure, the pharmaceutically acceptable carrier may include gum arabic, animal oil, benzyl alcohol, benzyl benzoate, calcium stearate, carbomer, cetostearyl alcohol, cetyl alcohol, cholesterol, cyclodextrin, dextrose, diethanolamine, emulsifying wax, ethylene glycol stearate palmitate, glycerol, glyceryl monostearate, glyceryl stearate, glyceryl monooleate, glyceryl monostearate, hydrate, histidine, hydrochloric acid, hydroxypropyl cellulose, hydroxypropyl-β-cyclodextrin (HPBCD), hydroxypropyl methylcellulose (hydroxypropyl methylcellulose (HPMC)), lanolin, lanolin alcohol, lecithin, medium chain triglycerides, metal soap, methylcellulose, mineral oil, sodium monophosphate, monoethanolamine, oleic acid, polyethylene glycol (PEG 3350, PEG 4000, PEG 6000), polyoxyethylene-polyoxypropylene copolymer (poloxamer), polyoxyethylene alkyl ether, polyoxyethylene castor oil, polyoxyethylene castor oil derivative, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene stearate, polysorbate, polyoxyethylene (20) sorbitan monolaurate (Tween 20, polysorbate 20), polyoxyethylene (20) sorbitan monooleate (Tween 80, polysorbate 80), povidone, propylene glycol alginate, saline, sodium chloride, sodium citrate, sodium citrate dihydrate, sodium hydroxide, sodium lauryl sulfate, sodium dihydrogen phosphate, disodium hydrogen phosphate, sorbitan esters, stearic acid, stearyl alcohol, sunflower oil, tragacanth gum, triethanolamine, vegetable oil, water, xanthan gum, or a combination thereof.

[0148] In some embodiments, the pharmaceutically acceptable carrier includes dextrose, glycerol, histidine, hydrochloric acid, hydroxypropyl cellulose, hydroxypropyl-β-cyclodextrin (HPBCD), hydroxypropyl methylcellulose (hydroxypropyl methylcellulose (HPMC)), polyoxyethylene (20) sorbitan monolaurate (Tween 20, polysorbate 20), polyethylene glycol (PEG 3350, PEG 4000, PEG 6000), polyoxyethylene-polyoxypropylene copolymer (poloxamer 188, poloxamer 407), polyoxyethylene (20) sorbitan monooleate (Tween 80, polysorbate 80), saline, sodium chloride, sodium citrate, sodium citrate dihydrate, sodium lauryl sulfate, sodium dihydrogen phosphate, disodium hydrogen phosphate, or a combination thereof.

[0149] The present disclosure also relates to a method for treating a disease or condition in a subject in need thereof, the method comprising administering to the subject a therapeutically effective dose of a compound or pharmaceutical composition disclosed herein.Any compound and pharmaceutical composition disclosed herein, or a combination thereof, may be suitable for treating a disease or condition.

[0150] Exemplary CNS diseases and conditions associated with GABA modulation include, but are not limited to, sleep disorders (e.g., insomnia), mood disorders (e.g., depression, dysthymic disorder (e.g., minor depression), bipolar disorder (e.g., I and / or II), anxiety disorders (e.g., generalized anxiety disorder (GAD), social anxiety disorder), stress, post-traumatic stress disorder (PTSD), obsessive-compulsive disorder (e.g., obsessive-compulsive disorder (OCD)), schizophrenia spectrum disorders (e.g., schizophrenia, schizoaffective disorder), convulsive disorders (e.g., epilepsy (e.g., status epilepticus (SE)), seizures), memory and / or cognitive disorders (e.g., attention disorders (e.g., attention deficit hyperactivity disorder (ADHD)), dementia (e.g., Alzheimer's type dementia, Lewis body type dementia, vascular dementia), movement disorders (e.g., Huntington's disease, Parkinson's disease, In some embodiments, the CNS disease or condition is a combination of autism, autism spectrum disorder (ASD), a personality disorder (e.g., antisocial personality disorder, obsessive-compulsive personality disorder), autism spectrum disorder (ASD) (e.g., autism, monogenic causes of autism, such as synaptic dysfunction, e.g., Rett syndrome, fragile X syndrome, Angelman syndrome), pain (e.g., neuropathic pain, injury-related pain syndrome, acute pain, chronic pain), traumatic brain injury (TBI), vascular disease (e.g., stroke, ischemia, vascular malformation), substance abuse disorder and / or withdrawal syndrome (e.g., opioid, cocaine and / or alcohol addiction), tinnitus, or a combination thereof. In some embodiments, the CNS disease or condition is CDD, MDD, PPD, essential tremor, PTSD, SE, ESE, fragile X syndrome, Parkinson's disease, or treatment-resistant depression. In some embodiments, the CNS disease or condition is CDD, MDD, PPD, excessive tremor, PTSD, SE, ESE, or fragile X syndrome.

[0151] In some embodiments of the methods disclosed herein, the disease or condition comprises a sleep disorder, insomnia, a mood disorder, depression, dysthymic disorder, minor depression, bipolar disorder, anxiety disorder, generalized anxiety disorder (GAD), social anxiety disorder, stress, post-traumatic stress disorder (PTSD), obsessive compulsive disorder, obsessive compulsive disorder (OCD), schizophrenia spectrum disorder, schizophrenia, schizoaffective disorder, convulsive disorder, epilepsy, status epilepticus (SE), seizures, memory and / or cognitive disorders, attention disorders, attention deficit hyperactivity disorder (ADHD), dementia, Alzheimer's dementia, dementia with Lewy bodies, vascular dementia, movement disorders, Huntington's disease, Parkinson's disease, personality disorders, antisocial personality disorder, obsessive-compulsive personality disorder, autism spectrum disorder (ASD), autism, monogenic causes of autism, synaptic dysfunction, Rett syndrome, fragile X syndrome, Angelman syndrome, neuropathic pain, injury-related pain syndrome, acute pain, chronic pain, traumatic brain injury (TBI), vascular disease, stroke, ischemia, vascular malformation, substance abuse disorder and / or withdrawal syndrome, opioid addiction (addition to opiates), cocaine addiction (addition to cocaine), alcohol addiction (addition to alcohol), tinnitus, or a combination thereof.

[0152] In some embodiments of the present method, the disorder is anxiety, major depressive disorder, postpartum disorder, Alzheimer's disease, Parkinson's disease, epilepsy, focal seizures, PCDH19 pediatric epilepsy, pediatric genetic epilepsy, CDKL5 deficiency (CDD), catamenial epilepsy, infantile spasms, fragile X syndrome, depression, postpartum depression, or premenstrual syndrome.

[0153] In some embodiments, the present disclosure relates to the use of a neuroactive steroid disclosed herein for the manufacture of a medicament for treating a disease, wherein the disease comprises a sleep disorder, insomnia, a mood disorder, depression, dysthymic disorder, minor depression, bipolar disorder, anxiety disorder, generalized anxiety disorder (GAD), social anxiety disorder, stress, post-traumatic stress disorder (PTSD), obsessive compulsive disorder, obsessive compulsive disorder (OCD), schizophrenia spectrum disorder, schizophrenia, schizoaffective disorder, convulsive disorder, epilepsy, status epilepticus (SE), seizures, memory and / or cognitive disorders, attention disorders, attention deficit hyperactivity disorder (ADHD), DHD), dementia, Alzheimer's type dementia, dementia with Lewy bodies, vascular dementia, movement disorders, Huntington's disease, Parkinson's disease, personality disorders, antisocial personality disorder, obsessive-compulsive personality disorder, autism spectrum disorder (ASD), autism, monogenic causes of autism, synaptic dysfunction, Rett syndrome, fragile X syndrome, Angelman syndrome, neuropathic pain, injury-related pain syndrome, acute pain, chronic pain, traumatic brain injury (TBI), vascular disease, stroke, ischemia, vascular malformation, substance abuse disorder and / or withdrawal syndrome, opioid addiction, cocaine addiction, alcohol addiction, tinnitus, or a combination thereof.

[0154] Any of the neuroactive steroids disclosed herein, or combinations thereof, may be useful in treating the diseases and conditions described above.

[0155] The pharmaceutical composition can be administered to a subject by intramuscular (IM) injection, subcutaneous (SC) injection, intravenous (IV) injection, oral administration, topical administration, implantation administration, or a combination thereof.

[0156] In some embodiments, the NAS compounds disclosed herein have excellent pharmaceutical properties.

[0157] The present disclosure will now be illustrated in the following non-limiting examples.

[0158] Example

[0159] The present invention is further defined in the following examples. It should be understood that these examples, while indicating preferred embodiments of the present invention, are given by way of illustration only. From the above discussion and these examples, one skilled in the art will be able to ascertain the essential characteristics of the present invention and, without departing from the spirit and scope of the present invention, may make various changes and modifications to adapt the present invention to various uses and conditions.

[0160] Example 1: (3R,5S,8R,9S,10S,13S,14S,17S)-3,10,13-trimethyl-17-(3-methyloxetane-3-yl)-1,2,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydrocyclopenta[a]phenanthrene-3-ol

[0161]

[0162] Preparation of (3R,5S,8R,9S,10S,13S,14S,17S)-17-(2-methoxy-1-methyl-vinyl)-3,10, 1,2,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydrocyclopenta[a]phenanthrene-3-ol

[0163]

[0164] To a solution of 1-[(3R,5S,8R,9S,10S,13S,14S,17S)-3-hydroxy-3,10,13-trimethyl-1,2,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydrocyclopenta[a]phenanthren-17-yl]ethanone (1.00 g, 3.01 mmol, 1 eq) and methoxymethyl(triphenyl)phosphonium chloride (1.34 g, 3.91 mmol, 1.3 eq) in THF (10 mL) was added potassium 2-methylpropan-2-olate (439 mg, 3.91 mmol, 1.3 eq). The mixture was stirred at 20 ° C for 16 h. The reaction mixture was quenched with H O (10 mL) and extracted with DCM (10 mL x 3). The combined organic layers were washed with H2O (20 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography ( 12g S Purification on a silica gel flash column with a gradient of 0-30% ethyl acetate in petroleum ether at 25 mL / min gave (3R,5S,8R,9S,10S,13S,14S,17S)-17-(2-methoxy-1-methyl-vinyl)-3,10,13-trimethyl-1,2,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydrocyclopenta[a]phenanthrene-3-ol (950 mg, 87.6% yield) as a white solid as a mixture of E and Z isomers in a ratio of ~3:1. The major isomer 1 H NMR (400MHz, CDCl3) δ (ppm) 5.79 (s, 1H), 3.57 (s, 3H), 1.95-0.85 (m, 28H), 0.82-0.70 (m, 4H), 0.56 (s, 3H).

[0165] Preparation of 2-[(3R,5S,8R,9S,10S,13S,14S,17R)-3-hydroxy-3,10,13-trimethyl-1,2,4,5,6, 7,8,9,11,12,14,15,16,17-Tetradecahydrocyclopenta[a]phenanthren-17-yl]propanal

[0166]

[0167] To a solution of (3R,5S,8R,9S,10S,13S,14S,17S)-17-(2-methoxy-1-methyl-vinyl)-3,10,13-trimethyl-1,2,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydrocyclopenta[a]phenanthren-3-ol (950 mg, 2.63 mmol, 1 eq) in THF (5 mL) was added HCl (510 mg, 5.18 mmol, 0.5 mL, 37% in H2O, 1.96 eq). The mixture was stirred at 20 °C for 0.5 h. The reaction mixture was neutralized with saturated Na2CO3 to pH ~7 and then extracted with DCM (10 mL x 3). The combined organic layers were washed with H2O (10 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography ( 4g The product was purified by silica gel flash column (eluting with a gradient of 0-20% ethyl acetate in petroleum ether at 25 mL / min) to afford 2-[(3R,5S,8R,9S,10S,13S,14S,17R)-3-hydroxy-3,10,13-trimethyl-1,2,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydrocyclopenta[a]phenanthren-17-yl]propanal (900 mg, 98.6% yield) as a white solid. 1 HNMR (400MHz, CDCl3) δ (ppm) 9.53 (d, J = 5.2Hz, 1H), 2.42-2.24 (m, 1H), 1.96-1.78 (m, 1H), 1.70-0.76 (m, 27H), 0.86-0.70 (m, 4H), 0.67 (s, 3H).

[0168] Preparation of 2-[(3R,5S,8R,9S,10S,13S,14S,17S)-3-hydroxy-3,10,13-trimethyl-1,2,4,5,6, 7,8,9,11,12,14,15,16,17-Tetradecahydrocyclopenta[a]phenanthren-17-yl]-2-methyl-propane-1,3-diol

[0169]

[0170] A mixture of 2-[(3R,5S,8R,9S,10S,13S,14S,17R)-3-hydroxy-3,10,13-trimethyl-1,2,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydrocyclopenta[a]phenanthren-17-yl]propanal (400 mg, 1.15 mmol, 1 eq), HCHO (8.72 g, 107.45 mmol, 8.00 mL, 37%, 93.09 eq), K2CO3 (638.10 mg, 4.62 mmol, 4 eq) in H2O (5 mL) and EtOH (5 mL) was degassed and purged with N2 three times, and then the mixture was stirred at 100 °C under N2 atmosphere for 16 h. The suspension was filtered and the resulting residue was washed with H2O (10 mL) to give a crude product of 2-[(3R,5S,8R,9S,10S,13S,14S,17S)-3-hydroxy-3,10,13-trimethyl-1,2,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydrocyclopenta[a]phenanthrene-17-yl]-2-methyl-propane-1,3-diol as a white solid (390 mg, 89.3% yield). 1 H NMR (400MHz, CD3OD) δ (ppm) 3.68-3.51 (m, 2H), 3.37 (m, 2H), 2.02-1.89 (m, 1H), 1.76-0.67 (m, 34H).

[0171] Preparation of (3R,5S,8R,9S,10S,13S,14S,17S)-3,10,13-trimethyl-17-(3-methyloxetane) alkyl-3-yl)-1,2,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydrocyclopenta[a]phenanthrene-3-ol

[0172]

[0173] To a solution of 2-[(3R,5S,8R,9S,10S,13S,14S,17S)-3-hydroxy-3,10,13-trimethyl-1,2,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydrocyclopenta[a]phenanthren-17-yl]-2-methyl-propane-1,3-diol (200 mg, 0.528 mmol, 1 eq) in THF (3 mL) was added NaH (25.4 mg, 0.634 mmol, 60% in mineral oil, 1.2 eq) and the mixture was stirred at 20 °C for 0.5 h. Then p-Toluenesulfonyl chloride (101 mg, 0.528 mmol, 1 eq) was added and the mixture was stirred at 20 ° C for 1 h, and then another portion of NaH (25.4 mg, 0.634 mmol, 60%, 1.2 eq) was added. The resulting mixture was stirred for another 16 h at 20 ° C. The mixture was then quenched with water (3 mL) and extracted with EtOAc (5 mL x 3). The combined organic layer was dried over Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 4g The product was purified by flash column chromatography on silica gel (eluted with a gradient of 0-40% ethyl acetate / petroleum ether at 20 mL / min) to give the product (60 mg, 31% yield). The product was then recrystallized from EtOAc (2 mL) to give (3R, 5S, 8R, 9S, 10S, 13S, 14S, 17S)-3,10,13-trimethyl-17-(3-methyloxetane-3-yl)-1,2,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydrocyclopenta[a]phenanthrene-3-ol (10 mg, 5.17% yield, 98.5% purity) as colorless crystals. LCMS (ESI) m / z, C 24 H 40 O2: Calculated 360.3, Found [M-OH] + :343.3. 1 H NMR (400MHz, CDCl3) δ (ppm) 4.85 (d, J = 6.0Hz, 1H), 4.59 (d, J = 5.2Hz, 1H), 4.20 (d, J = 6.0Hz, 1H), 4.14 (d, J=5.2Hz,1H),2.09-1.79(m,3H),1.76-1.63(m,2H),1.61-0.82(m,23H),0.80-0.66(m,4H),0.53(s,3H). 13C NMR(100MHz,CDCl3)δ(ppm)83.55,79.93,69.78,56.38,55.26,54.04,43.46,41.93,41.77, 41.08,39.86,35.51,35.03,34.86,31.91,28.38,26.36,24.30,24.24,20.73,12.38,11.19.

[0174] Example 2: (3R,5R,8R,9R,10S,13S,14S,17S)-3,13-dimethyl-17-(3-methyloxetane-3-yl)-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0175]

[0176] The compound (3R,5R,8R,9R,10S,13S,14S,17S)-3,13-dimethyl-17-(3-methyloxetan-3-yl)-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetrahydro-1H-cyclopenta[a]phenanthrene-3-ol was prepared using the same reaction sequence as for the preparation of (3R,5S,8R,9S,10S,13S,14S,17S)-3,10,13-trimethyl-17-(3-methyloxetan-3-yl)-1,2,4,5,6,7,8,9,11,12,14,15,16,17-tetrahydro-1H-cyclopenta[a]phenanthrene-3-ol. The difference was that 1-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl]ethanone (23 mg, 23.7% yield, 98% purity, white solid) was used instead of 1-[(3R,5S,8R,9S,10S,13S,14S,17S)-3-hydroxy-3,10,13-trimethyl-1,2,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydrocyclopenta[a]phenanthrene-17-yl]ethanone. LCMS (ESI) m / z, C 23 H 38 O2: Calculated 346.3, Found [M-OH] + :329.3. 1H NMR (400MHz, CDCl3) 4.85 (d, J = 6.0 Hz, 1H), 4.59 (d, J = 5.2 Hz, 1H), 4.21 (d, J = 6.0 Hz, 1H), 4.14 (d, J = 5.2 Hz, 1H), 2.05-0.86 (m, 30H), 0.54 (s, 3H). 13 C NMR (100MHz, CDCl3) δ (ppm) 83.58, 79.85, 72.07, 55.40, 43.61, 41.93, 41.20, 39. 90,37.55,34.71,34.50,31.40,26.45,25.99,25.55,25.38,24.39,24.13,12.35.

[0177] Example 3: (3R,5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(2-methyloxetane-2-yl)-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0178]

[0179] Preparation of (3R,5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(2-methyloxirane-2- (4-Hydroxy)-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0180]

[0181] A mixture of potassium tert-butoxide (2.11 g, 18.8 mmol, 3 eq) and trimethylsulfoxide iodide (4.15 g, 18.8 mmol, 3 eq) in t-BuOH (25 mL) was stirred at 70 ° C for 1 h, and then 1-[(3R, 5S, 8R, 9S, 10S, 13S, 14S, 17S)-3-hydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl]ethanone (2.00 g, 6.28 mmol, 1 eq) was added. The resulting mixture was stirred at 70 ° C for another 50 h, and then concentrated. The residue was purified by flash silica gel chromatography ( 20g The product was purified by silica gel flash column (eluting with a gradient of 0-10% ethyl acetate in petroleum ether at 20 mL / min) to afford (3R,5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(2-methyloxiran-2-yl)-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol (900 mg, 43% yield) as a white solid.

[0182] Preparation of (3R,5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(2-methyloxetane- 2-yl)-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0183]

[0184] To a solution of trimethylsulfoxide iodide (596 mg, 2.71 mmol, 3 eq) in DMSO (5 mL) was added NaH (180 mg, 4.51 mmol, 60% in mineral oil, 5 eq). The mixture was stirred at room temperature for 1 h, then (3R, 5S, 8R, 9S, 10S, 13S, 14S, 17S)-10,13-dimethyl-17-(2-methyloxetan-2-yl)-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol (299 mg, 0.90 mmol, 1 eq) was added and the resulting mixture was stirred for an additional 16 h. The mixture was diluted with water (15 mL) and extracted with EtOAc (15 mL x 2). The organic layers were combined, washed with brine (20 mL), dried (Na2SO4), filtered and concentrated under reduced pressure. The resulting residue was purified by flash silica gel chromatography ( 4g Silica gel flash column, eluted with 0-10% ethyl acetate / petroleum ether gradient at 15 mL / min) was purified. The product (combined with another batch) was purified again by preparative TLC (dichloromethane / ethyl acetate=30 / 1) to give (3R, 5S, 8R, 9S, 10S, 13S, 14S, 17S)-10,13-dimethyl-17-(2-methyloxetane-2-yl)-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol (19.8 mg, 3.1% yield) as a white solid. LCMS (ESI) m / z, C 23 H 38 O2: Calculated 346.29, Found [M-OH] + :329.28. 1HNMR (400MHz, CDCl3) δ (ppm) 4.54-4.50 (m, 1H) 4.37-4.35 (m, 1H), 4.05 (s, 1H), 2.63-2.56 (m, 1H), 2.20-2. 11(m,2H),2.04-2.01(m,1H),1.89-1.88(m,1H),1.69-1.67(m,4H),1.57-0.88(m,19H),0.78-0.70(m,7H). 13 C NMR (100MHz, CDCl3) δ (ppm) 89.02, 66.58, 64.59, 59.42, 56.81, 54.22, 43.00, 39.96, 39.11, 36. 06,35.85,34.90,33.41,32.15,31.89,28.99,28.52,28.32,23.85,22.85,20.50,12.65,11.17.

[0185] Example 4: (3R,5R,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(2-methyloxetane-2-yl)-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0186]

[0187] Preparation of (3R,5R,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(2-methyloxirane-2- (4-Hydroxy)-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0188]

[0189] To a solution of potassium tert-butoxide (2.11 g, 18.8 mmol, 3 eq) in t-BuOH (25 mL) was added trimethylsulfoxide iodide (4.15 g, 18.8 mmol, 3 eq). The mixture was stirred at 40 ° C for 1 h, then 1-[(3R, 5R, 8R, 9S, 10S, 13S, 14S, 17S)-3-hydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl]ethanone (2.00 g, 6.28 mmol, 1 eq) was added and the resulting mixture was stirred at 40 ° C for 40 h. The mixture was diluted with water (30 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were dried over Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 40g Purification by silica gel flash column, 0-35% ethyl acetate / petroleum ether gradient at 30 mL / min eluent afforded (3R,5R,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(2-methyloxiran-2-yl)-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol (750 mg, 36% yield) as a white solid.

[0190] Preparation of (3R,5R,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(2-methyloxetane- 2-yl)-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0191]

[0192] To a solution of potassium tert-butoxide (807 mg, 7.20 mmol, 6 eq) in t-BuOH (10 mL) was added trimethylsulfoxide iodide (1.58 g, 7.20 mmol, 6 eq), the mixture was stirred at 50° C. for 1 h, and then (3R, 5R, 8R, 9S, 10S, 13S, 14S, 17S)-10,13-dimethyl-17-(2-methyloxiran-2-yl)-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol (400 mg, 1.20 mmol, 1 eq) was added and the resulting mixture was stirred at 70° C. for 64 h. The mixture was diluted with water (30 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were dried over Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 20g Silica gel flash column, 0-35% ethyl acetate / petroleum ether gradient with 30mL / min eluent) to purify the product as a white solid (250mg, 60% yield). The product (150mg) was further purified by preparative TLC (dichloromethane / ethyl acetate = 5 / 1) to give (3R, 5R, 8R, 9S, 10S, 13S, 14S, 17S) -10,13-dimethyl -17- (2-methyloxetane -2- bases) -2,3,4,5,6,7,8,9,11,12,14,15,16,17- tetradecahydro -1H- cyclopenta [a] phenanthren-3-ol (20.7mg, 5.0% yield) as a white solid. LCMS (ESI) m / z, C23H38O2: calculated value 346.29, found value [M-OH]+: 329.28.1H NMR(400MHz, CDCl3)δ(ppm)4.55-4.50(m,1H),4.39-4.33(m,1H),3.68-3.61(m,1H),2.63-2.58(m,1H),2.21-2.09(m,2H),2.05-2.01(m,1H) ,1.92-1.76(m,4H),1.71-1.64(m,2H),1.57(s,3H),1.55-1.49(m,1H) ,1.46-1.38(m,7H),1.32-1.05(m,7H),1.01-0.92(m,4H),0.73(s,3H). 13C NMR (100MHz, CDCl3) δ (ppm) 88.99, 71.84, 64.60, 59.50, 56.80, 43.11, 42.04, 40.36, 40.14, 36. 41,35.31,35.27,34.56,33.42,30.51,28.33,27.16,26.32,23.93,23.37,22.94,20.56,12.61.

[0193] Example 5: (3R,5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(3-methyloxetane-3-yl)-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0194]

[0195] Preparation of (3R,5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(2-methyloxirane-2- (4-Hydroxy)-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0196]

[0197] A mixture of t-BuOK (3.17 g, 28.3 mmol, 3 eq) and trimethylsulfoxide iodide (6.22 g, 28.3 mmol, 3 eq) in t-BuOH (30 mL) was stirred at 50 ° C for 1 h, and then 1-[(3R, 5S, 8R, 9S, 10S, 13S, 14S, 17S)-3-hydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl]ethanone (3.00 g, 9.42 mmol, 1 eq) was added. The resulting mixture was stirred at 50 ° C for another 50 h, then concentrated and diluted with H2O (50 mL) and extracted with DCM (50 mL x 3). The combined organic layers were concentrated. The resulting residue was purified by flash silica gel chromatography ( 40g The product was purified by silica gel flash column (0-20% ethyl acetate / petroleum ether gradient at 35 mL / min eluent) to afford (3R,5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(2-methyloxiran-2-yl)-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol (2.10 g, 67% yield) as a white solid.

[0198] Preparation of 2-[(3R,5S,8R,9S,10S,13S,14S,17R)-3-hydroxy-10,13-dimethyl-2,3,4,5,6,7, 8,9,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propanal

[0199]

[0200] To a solution of (3R, 5S, 8R, 9S, 10S, 13S, 14S, 17S) -10, 13-dimethyl -17- (2-methyloxetane -2-yl) -2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetrahydro -1H- cyclopenta [a] phenanthren-3-ol (1.00 g, 3.01 mmol, 1 eq) in DCM (10 mL) was added BF3 Et2O (554 mg, 3.91 mmol, 0.48 mL, 1.3 eq) at 0 ° C. The resulting mixture was stirred at 0 ° C for 1 h, then diluted with H2O (30 mL) and extracted with DCM (30 mL x3). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, filtered and concentrated. The resulting residue was purified by flash silica gel chromatography ( 12g Purification by silica gel flash column, 0-20% ethyl acetate / petroleum ether gradient at 20 mL / min eluent afforded 2-[(3R,5S,8R,9S,10S,13S,14S,17R)-3-hydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propanal (550 mg, 55% yield) as a white solid. 1 H NMR (400MHz, CDCl3) δ (ppm) 9.57 (m, 1H), 4.05 (s, 1H), 2.36-2.26 (m, 1H), 1.94-1.81 (m, 1H), 1.69-0.93 (m, 24H), 0.79-0.66 (m, 7H).

[0201] Preparation of 3-hydroxy-2-[(3R,5S,8R,9S,10S,13S,14S,17S)-3-hydroxy-10,13-dimethyl-2,3, 4,5,6,7,8,9,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]-2-methyl-propanal

[0202]

[0203] A mixture of 2-[(3R,5S,8R,9S,10S,13S,14S,17R)-3-hydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propanal (480 mg, 1.44 mmol, 1 eq), HCHO (10.9 g, 134 mmol, 10.0 mL, 37% in H2O, 93 eq) and K2CO3 (199 mg, 1.44 mmol, 1 eq) in H2O (8 mL) and EtOH (8 mL) was degassed and purged with N2 three times, and then the mixture was stirred at 100 °C under N2 atmosphere for 16 h. The suspension was filtered, the filter cake was washed with H2O (10 mL), and the resulting residue was purified by flash silica gel chromatography ( 12g Purification by silica gel flash column, 0-10% methanol / dichloromethane gradient at 20 mL / min eluent afforded 3-hydroxy-2-[(3R,5S,8R,9S,10S,13S,14S,17S)-3-hydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]-2-methyl-propanal (300 mg, 57% yield) as a white solid.

[0204] Preparation of 2-[(3R,5S,8R,9S,10S,13S,14S,17S)-3-hydroxy-10,13-dimethyl-2,3,4,5,6,7, 8,9,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]-2-methyl-propane-1,3-diol

[0205]

[0206] To a mixture of 3-hydroxy-2-[(3R,5S,8R,9S,10S,13S,14S,17S)-3-hydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]-2-methyl-propanal (200 mg, 0.55 mmol, 1 eq) in THF (5 mL) and H2O (5 mL) was added HCHO (4.36 g, 53.3 mmol, 4.0 mL, 37% in H2O, 97 eq) and NaOH (88 mg, 2.21 mmol, 4 eq). The resulting reaction mixture was stirred at room temperature for 16 h, then concentrated and diluted with H2O (20 mL). The resulting mixture was filtered, and the filter cake was collected and washed with H2O (20 mL), then dried under vacuum to afford 2-[(3R,5S,8R,9S,10S,13S,14S,17S)-3-hydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]-2-methyl-propane-1,3-diol (190 mg, 94% yield) as a white solid.

[0207] Preparation of (3R,5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(3-methyloxetane- 3-yl)-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0208]

[0209] To a solution of 2-[(3R,5S,8R,9S,10S,13S,14S,17S)-3-hydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]-2-methyl-propane-1,3-diol (100 mg, 0.274 mmol, 1 eq) in DMF (5 mL) was added NaH (55 mg, 1.37 mmol, 60% in mineral oil, 5 eq). The mixture was stirred at room temperature for 0.5 h, then p-toluenesulfonyl chloride (57 mg, 0.301 mmol, 1.1 eq) was added. The resulting mixture was stirred at room temperature for 1 h, then another portion of NaH (10.9 mg, 0.274 mmol, 60% in mineral oil, 1 equivalent) was added, and the mixture was stirred at room temperature for another 16 h. The reaction mixture was then diluted with H2O (10 mL) and extracted with EtOAc (10 mL x 3). The combined organic layer was washed with brine (20 mL), dried over Na2SO4, filtered and concentrated. The crude product was merged with another batch, and the combined crude product was purified by flash silica gel chromatography ( 12g The product was purified by silica gel flash column with 0-20% ethyl acetate / petroleum ether gradient at 20 mL / min. The product was then recrystallized from EtOAc (10 mL) to give (3R, 5S, 8R, 9S, 10S, 13S, 14S, 17S) -10,13-dimethyl-17- (3-methyloxetane-3-yl) -2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetrahydro-1H-cyclopenta[a]phenanthrene-3-ol (67 mg, 32% yield) as a white solid. HRMS (ESI) m / z, C 23 H 38 O2: Calculated value 346.2875, experimental value (M+H) + :347.2948. 1 H NMR (400MHz, CDCl3) δ (ppm) 4.86-4.84 (d, J = 8.0Hz, 1H), 4.59-4.58 (d, J = 4.0Hz, 1H), 4.21-4.20 (d, J = 4.0Hz, 1H), 4.15-4.13 (d, J = 8.0Hz, 1H), 4.05 (s, 1H),2.05-1.82(m,3H),1.73-1.58(m,5H),1.54-1.43(m,8H),1.39-1.30(m ,3H),1.27-1.13(m,5H),1.10-0.89(m,2H),0.77-0.71(m,4H),0.53(s,3H). 13 C NMR (400MHz, CDCl3) δ (ppm) 83.58, 79.91, 66.54, 56.39, 55.26, 54.13, 43.45, 41.94, 39.85, 39. 07,36.04,35.83,34.98,32.12,31.91,28.97,28.47,26.34,24.30,24.21,20.50,12.39,11.16.

[0210] Example 6: (3R,5R,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(3-methyloxetane-3-yl)-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0211]

[0212] Preparation of 2-[(3R,5R,8R,9S,10S,13S,14S,17R)-3-hydroxy-10,13-dimethyl-2,3,4,5,6,7, 8,9,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propanal

[0213]

[0214] To a solution of (3R, 5R, 8R, 9S, 10S, 13S, 14S, 17S) -10, 13-dimethyl -17- (2-methyloxetane -2- bases) -2,3,4,5,6,7,8,9,11,12,14,15,16,17- tetradecahydro -1H- cyclopenta [a] phenanthren-3-ol (250 mg, 0.75 mmol, 1 equiv) in DCM (1 mL) was added B F3 · Et2O (149 mg, 1.05 mmol, 0.13 mL, 1.4 equiv). The resulting mixture was stirred at 20 ° C for 1.5 h. The mixture was quenched with saturated NaHCO3 (15 mL) and extracted with DCM (15 mL x3). The organic layer was washed with brine (20 mL), filtered and concentrated. The residue was purified by flash silica gel chromatography ( 4g Purification by silica gel flash column, 0-30% ethyl acetate / petroleum ether gradient at 25 mL / min eluent afforded 2-[(3R,5R,8R,9S,10S,13S,14S,17R)-3-hydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propanal (120 mg, 48% yield) as a white solid. 1HNMR (400MHz, CDCl3) δppm 9.58-9.53(m,1H),3.66-3.51(m,1H),2.38-2.30(m,1H),1.95-1.75(m,4H),1.71-1.59(m,3H),1.54-1.49(m,2H),1. 45-1.22(m,11H),1.13-1.10(m,4H),1.04(d,J=7.0Hz,2H),1.02-0.96(m,1H),0.94-0.92(m,3H),0.70-0.66(m,3H).

[0215] Preparation of 2-[(3R,5R,8R,9S,10S,13S,14S,17S)-3-hydroxy-10,13-dimethyl-2,3,4,5,6,7, 8,9,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]-2-methyl-propane-1,3-diol

[0216]

[0217] To a mixture of 2-[(3R,5R,8R,9S,10S,13S,14S,17R)-3-hydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl]propanal (120 mg, 0.36 mmol, 1 eq) in EtOH (2 mL) and H2O (2 mL) was added HCHO (2.73 g, 33.5 mmol, 2.5 mL, 37% in H2O, 93 eq) and K2CO3 (200 mg, 1.45 mmol, 4 eq). The resulting mixture was stirred at 100 ° C for 16 h and then concentrated. The residue was washed with water (4 mL) and dried. The resulting residue was purified by flash silica gel chromatography ( 4g The product was purified by silica gel flash column (eluting with a gradient of 0-10% ethyl acetate in petroleum ether at 25 mL / min) to afford 2-[(3R,5R,8R,9S,10S,13S,14S,17S)-3-hydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]-2-methyl-propane-1,3-diol (80 mg, 61% yield) as a white solid.

[0218] Preparation of (3R,5R,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(3-methyloxetane- 3-yl)-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0219]

[0220] To a solution of 2-[(3R,5R,8R,9S,10S,13S,14S,17S)-3-hydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]-2-methyl-propane-1,3-diol (80 mg, 0.22 mmol, 1 eq) in DMF (5 mL) at 25° C. was added NaH (35.1 mg, 0.88 mmol, 60% in mineral oil, 4 eq). The mixture was stirred at 25° C. for 30 min, then 4-methylbenzenesulfonyl chloride (46.0 mg, 0.24 mmol, 1.1 eq) was added. The resulting mixture was stirred at 25° C. for 64 h. The mixture was diluted with EtOAc (50 mL), washed with water (20 mL x 2), dried over Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 12g Purification by silica gel flash column, 0-20% ethyl acetate / petroleum ether gradient at 20 mL / min eluent afforded (3R,5R,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(3-methyloxetan-3-yl)-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol (32.0 mg, 42% yield) as a white solid. LCMS (ESI) m / z, C 23 H 38 O2: Calculated 346.29, found [M+H] + :347.29. 1 H NMR (400MHz, CDCl3) δ (ppm) 4.85 (d, J = 8.0Hz, 1H), 4.59 (d, J = 8.0Hz, 1H), 4.21 (d, J = 4.0Hz, 1H), 4.14 (d, J = 4.0Hz, 1H), 3.66 -3.61(m,1H),2.05-1.66(m,8H),1.57-1.45(m,4H),1.47-1.34(m,7H),1.31-1.07(m,7H),1.04-0.90(m,4H),0.52(s,3H). 13 C NMR(100MHz,CDCl3)δ(ppm)83.59,79.88,71.79,56.39,55.35,43.55,42.00,41.93,40.31, 40.03,36.38,35.35,35.28,34.54,30.48,27.10,26.36,24.41,24.30,23.32,20.56,12.35.

[0221] Example 7: (3R,5R,8R,9R,10S,13S,14S,17S)-3,13-dimethyl-17-(3-methyltetrahydrofuran-3-yl)-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0222]

[0223] Preparation of 2-cyano-2-((3R,5R,8R,9R,10S,13S,14S)-3-hydroxy-3,13-dimethylhexadecahydro-17 H - Ethyl cyclopenta[a]phenanthren-17-ylidene) acetate

[0224]

[0225] To a mixture of (3R,5R,8R,9R,10S,13S,14S)-3-hydroxy-3,13-dimethyl-1,2,4,5,6,7,8,9,10,11,12,14,15,16-tetradecahydrocyclopenta[a]phenanthrene-17-one (7.00 g, 24.1 mmol, 1 eq) in toluene (200 mL) was added ethyl 2-cyanoacetate (13.6 g, 120 mmol, 5 eq), NHOAc (5.57 g, 72.3 mmol, 3 eq) and HOAc (26.2 g, 436 mmol, 25 mL, 18.1 eq). The resulting mixture was stirred at 135° C. under a Dean-Stark water separator for 16 h. The reaction mixture was then concentrated and diluted with EtOAc (400 mL), washed with saturated aqueous NaHCO3 (200 mL x 2) and brine (200 mL), and the organic layer was dried over Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 220g The product was purified by silica gel flash column (eluting with a 0-10% ethyl acetate in dichloromethane gradient at 80 mL / min) to afford ethyl 2-cyano-2-((3R,5R,8R,9R,10S,13S,14S)-3-hydroxy-3,13-dimethylhexahydro-17H-cyclopenta[a]phenanthren-17-ylidene)acetate (7.00 g, 73.3% yield) as a white solid.

[0226] Preparation of 2-cyano-2-[(3R,5R,8R,9R,10S,13S,14S,17R)-3-hydroxy-3,13-dimethyl-2,4,5, 6,7,8,9,10,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]acetic acid ethyl ester

[0227]

[0228] To a solution of ethyl 2-cyano-2-((3R,5R,8R,9R,10S,13S,14S)-3-hydroxy-3,13-dimethylhexahydro-17H-cyclopenta[a]phenanthrene-17-ylidene)acetate (7.02 g, 18.2 mmol, 1 equiv) in EtOH (25 mL) and THF (75 mL) was added Pd / C (800 mg, 10 wt% loading), and the mixture was stirred under H (15 psi) at 20 °C for 6 h. The resulting reaction mixture was filtered and the filtrate was concentrated to give ethyl 2-cyano-2-[(3R,5R,8R,9R,10S,13S,14S,17R)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetrahydro-1H-cyclopenta[a]phenanthren-17-yl]acetate (6.84 g, 96.8% yield) as a white solid, which was used directly in the next step without further purification.1 H NMR (400MHz, CDCl3) δ (ppm) 4.28-4.23 (m, 2H) 3.41-3.26 (m, 1H) 2.21-2.05 (m, 2H) 1.81-1.79 (m, 6H) 1.42-1.06 (m, 22H) 0.77-0.76 (d, J = 4.0Hz, 3H).

[0229] Preparation of 2-cyano-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5, Ethyl 6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propionate

[0230]

[0231] To a solution of ethyl 2-cyano-2-[(3R,5R,8R,9R,10S,13S,14S,17R)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl]acetate (500 mg, 1.29 mmol, 1 eq) in DMF (5 mL) was added KCO (356 mg, 2.58 mmol, 2 eq) and MeI (915 mg, 6.45 mmol, 5 eq). The resulting mixture was stirred at room temperature for 3 h, then diluted with EtOAc (15 mL), washed with water (10 mL x3), brine (10 mL). The organic layer was dried over NaSO, filtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography ( 4g The product was purified by silica gel flash column (eluting with a gradient of 0-30% ethyl acetate in petroleum ether at 20 mL / min) to afford ethyl 2-cyano-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propanoate (450 mg, 86.9% yield) as a white solid.

[0232] Preparation of 3-hydroxy-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5, 6,7,8,9,10,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]-2-methyl-propionitrile

[0233]

[0234] To a mixture of ethyl 2-cyano-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl]propanoate (430 mg, 1.07 mmol, 1 eq) and NaBH4 (486 mg, 12.8 mmol, 12 eq) in THF (4 mL) was added MeOH (2 mL). The resulting mixture was stirred at 75 ° C for 16 h. The reaction mixture was quenched with water (8 mL) and extracted with EtOAc (10 mL x2). The combined organic layers were dried over Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 4g The product was purified by silica gel flash column (eluting with a gradient of 0-80% ethyl acetate in petroleum ether at 20 mL / min) to afford 3-hydroxy-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]-2-methyl-propionitrile (360 mg, 93.5% yield) as a white solid.

[0235] Preparation of 2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8, 9,10,11,12,14,15,16,17-Tetrahydro-1H-cyclopenta[a]phenanthren-17-yl]-2-methyl-3-oxo-propionitrile

[0236]

[0237] To a solution of 3-hydroxy-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl]-2-methyl-propionitrile (200 mg, 0.56 mmol, 1 eq) in DCM (3 mL) was added Dess-Martin periodinane (306 mg, 0.72 mmol, 1.3 eq). The resulting mixture was stirred at room temperature for 2 h. The reaction mixture was quenched with saturated Na2SO3 (3 mL) and then extracted with DCM (5 mL x 2). The combined organic layers were washed with H2O (10 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography ( 4g The product was purified by silica gel flash column (eluting with a gradient of 0-50% ethyl acetate in petroleum ether at 20 mL / min) to afford 2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]-2-methyl-3-oxo-propionitrile (180 mg, 90.5% yield) as a white solid.

[0238] Preparation of 2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8, 9,10,11,12,14,15,16,17-Tetrahydro-1H-cyclopenta[a]phenanthren-17-yl]-2-methyl-but-3-enenitrile

[0239]

[0240] A mixture of t-BuOK (113 mg, 1.01 mmol, 2 eq) and methyl(triphenyl)phosphonium bromide (360 mg, 1.01 mmol, 2 eq) in THF (5 mL) was stirred at 30 ° C for 1 h, and then a mixture of 2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetrahydro-1H-cyclopenta[a]phenanthren-17-yl]-2-methyl-3-oxo-propionitrile (180 mg, 0.50 mmol, 1 eq) in THF (3 mL) was added. The resulting mixture was stirred at 30 ° C for another 2 h, then diluted with EtOAc (15 mL) and washed with water (5 mL x 3). The organic layer was washed with brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 4g The product was purified by silica gel flash column (eluting with a gradient of 0-25% ethyl acetate in petroleum ether at 20 mL / min) to afford 2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]-2-methyl-but-3-enenitrile (160 mg, 89.4% yield) as a white solid. 1H NMR (400MHz, CDCl3) δ (ppm) 5.70-5.63 (m, 1H), 5.56-5.52 (d, J = 17.2Hz, 1H), 5.18-5.16 ( d,J=10.0Hz,1H),1.98-1.79(m,8H),1.65-1.55(m,3H),1.44-0.99(m,20H),0.86(s,3H).

[0241] Preparation of 2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8, 9,10,11,12,14,15,16,17-Tetrahydro-1H-cyclopenta[a]phenanthren-17-yl]-2-methyl-but-3-enal

[0242]

[0243] To a solution of 2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl]-2-methyl-but-3-enenitrile (120 mg, 0.34 mmol, 1 eq) in toluene (3 mL) was added DIBAL-H (1.0 M in toluene, 1.01 mL, 3 eq). The resulting mixture was stirred at 0 °C for 3 h, then quenched with saturated aqueous NH4Cl solution (3 mL) and extracted with DCM (5 mL x 3). The organic layers were combined, washed with brine (10 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography ( 4g The product was purified by silica gel flash column (eluting with a gradient of 0-20% ethyl acetate in petroleum ether at 20 mL / min) to afford 2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]-2-methyl-but-3-enal (100 mg, 82.6% yield) as a white solid.

[0244] Preparation of (3R,5R,8R,9R,10S,13S,14S,17S)-17-[1-(hydroxymethyl)-1-methyl-allyl]-3,13- Dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0245]

[0246] To a solution of 2-[(3R, 5R, 8R, 9R, 10S, 13S, 14S, 17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl]-2-methyl-but-3-enal (110 mg, 0.31 mmol, 1 equiv) in THF (2 mL) was added NaBH4 (58.0 mg, 1.55 mmol, 5 equiv). The resulting mixture was stirred at 20 ° C for 2 h. The reaction mixture was then diluted with water (2 mL) and extracted with EtOAc (5 mL x 2). The organic layers were combined, washed with brine, dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography ( 4g The product was purified by silica gel flash column (eluting with a gradient of 0-40% ethyl acetate in petroleum ether at 20 mL / min) to afford (3R,5R,8R,9R,10S,13S,14S,17S)-17-[1-(hydroxymethyl)-1-methyl-allyl]-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol (70 mg, 63.3% yield) as a white solid.

[0247] Preparation of 2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]-2-methyl-butane-1,4-diol

[0248]

[0249] To a solution of (3R,5R,8R,9R,10S,13S,14S,17S)-17-[1-(hydroxymethyl)-1-methyl-allyl]-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol (40 mg, 0.11 mmol, 1 eq) in THF (3 mL) was added BH3·THF (1.0 M in THF, 0.55 mL, 5 eq) at 0° C. The resulting mixture was stirred at 20° C. for 4 h and then quenched with H2O (0.5 mL). To the resulting mixture was added NaOH (3.0 M in H2O, 0.4 mL, 11 eq) and H2O2 (472 mg, 4.16 mmol, 0.4 mL, 30% in H2O, 38 eq) in sequence, and the mixture was stirred at 20°C for another 16 h. The resulting mixture was diluted with saturated Na2SO3 (2 mL) and extracted with EtOAc (8 mL x 3). The organic layers were combined, washed with brine (10 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC (petroleum ether / ethyl acetate = 1 / 1) to give 2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetrahydro-1H-cyclopenta[a]phenanthren-17-yl]-2-methyl-butane-1,4-diol (20 mg, 47.6% yield) as a colorless oil. 1 H NMR (400MHz, CDCl3) δ (ppm) 3.73-3.66 (m, 3H), 3.51-3.49 (d, J = 11.2Hz, 1H), 2.73 (s, 1H), 1.96-0.94 (m, 33H), 0.79 (s, 3H).

[0250] Preparation of (3R,5R,8R,9R,10S,13S,14S,17S)-3,13-dimethyl-17-(3-methyltetrahydrofuran-3- 1H-cyclopenta[a]phenanthrene-3-ol

[0251]

[0252] To a solution of 2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]-2-methyl-butane-1,4-diol (20 mg, 0.053 mmol, 1 eq) in DMF (1 mL) was added NaH (21.1 mg, 0.53 mmol, 60% in mineral oil, 10 eq). The mixture was stirred at 20 ° C for 1 h, then 4-methylbenzenesulfonyl chloride (15.1 mg, 0.079 mmol, 1.5 eq) was added and the mixture was stirred for an additional 2 h. The reaction mixture was then quenched with water (1 mL) and extracted with EtOAc (5 mL x 2). The combined organic layers were washed with H2O (3 mL), brine (3 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by preparative TLC (petroleum ether / ethyl acetate = 2 / 1) to give (3R, 5R, 8R, 9R, 10S, 13S, 14S, 17S) -3,13-dimethyl-17- (3-methyltetrahydrofuran-3-yl) -2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetrahydro-1H-cyclopenta [a] phenanthren-3-ol (6.1 mg, 32.0% yield) as a white solid. LCMS (ESI) m / z, C 24 H 40 O2: Calculated 360.57, Found [M-OH] + :343.3. 1 H NMR (400MHz, CDCl3) δ (ppm) 3.89-3.79 (m, 2H) 3.52-3.49 (m, 2H) 1.86-1.79 (m, 5H) 1.64-1.61 (m, 5H) 1.42-1.07 (m, 23H) 0.74 (s, 3H). 13 CNMR(100MHz,CDCl3)δ(ppm)79.39,72.05,67.42,57.80,55.28,45.54,43.91,41.27,41.21,40.36, 40.06,38.20,37.72,34.76,34.57,31.46,26.44,25.97,25.54,25.43,23.94,23.87,23.83,14.27.

[0253] Example 8: (3R,5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(2-methyltetrahydrofuran-2-yl)-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0254]

[0255] Preparation of (3R,5S,8R,9S,10S,13S,14S,17S)-17-(1-hydroxy-1-methyl-but-3-enyl)-10,13- Dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0256]

[0257] To a solution of 1-[(3R,5S,8R,9S,10S,13S,14S,17S)-3-hydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl]ethanone (1.00 g, 3.14 mmol, 1 eq) in THF (15 mL) was added allyl(bromide)magnesium (1.0 M in ether, 5.0 mL, 1.6 eq) dropwise at 0 ° C. The resulting mixture was allowed to warm to room temperature and stirred for 20 h. The reaction mixture was then diluted with water (15 mL) and extracted with EtOAc (20 mL x 2). The organic layers were combined, washed with brine (10 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography ( 12g The product was purified by silica gel flash column (eluting with a gradient of 0-10% ethyl acetate in petroleum ether at 20 mL / min) to afford (3R,5S,8R,9S,10S,13S,14S,17S)-17-(1-hydroxy-1-methyl-but-3-enyl)-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol) as a white solid (340 mg, 30.0% yield).

[0258] Preparation of 4-[(3R,5S,8R,9S,10S,13S,14S,17S)-3-hydroxy-10,13-dimethyl-2,3,4,5,6,7, 8,9,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]pentane-1,4-diol

[0259]

[0260] To a solution of (3R,5S,8R,9S,10S,13S,14S,17S)-17-(1-hydroxy-1-methyl-but-3-enyl)-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol (130 mg, 0.36 mmol, 1 eq) in THF (5 mL) at 0°C was added BH3·THF (1.0 M in THF, 1.08 mL, 3 eq). The resulting mixture was stirred at room temperature for 3 h, then NaOH (3.0 M in water, 1.32 mL, 11 eq) and H2O2 (1.53 g, 13.5 mmol, 1.38 mL, 30% in H2O, 37.5 eq) were added, and the mixture was stirred at room temperature for another 16 h. The reaction was quenched by the addition of saturated aqueous Na2SO3 (5 mL) and extracted with EtOAc (15 mL x 3). The combined organic layers were washed with water (100 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give 4-[(3R,5S,8R,9S,10S,13S,14S,17S)-3-hydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl]pentane-1,4-diol (110 mg, crude) as a white solid which was used in the next step without further purification.

[0261] Preparation of (3R,5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(2-methyltetrahydrofuran-2- (4-Hydroxy)-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0262]

[0263] To a solution of 4-[(3R,5S,8R,9S,10S,13S,14S,17S)-3-hydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]pentane-1,4-diol (100 mg, 0.26 mmol, 1 eq) in THF (5 mL) was added NaH (31.7 mg, 0.79 mmol, 60% in mineral oil, 3 eq). The mixture was stirred at 25 °C for 1 h before the addition of 4-methylbenzenesulfonyl chloride (59.3 mg, 0.31 mmol, 1.2 eq). The resulting mixture was stirred at 25 ° C for another 1 h, then another portion of NaH (31.7 mg, 0.79 mmol, 60% in mineral oil, 3 eq) was added, and the mixture was stirred at 25 ° C for another 48 h. The reaction mixture was quenched with water (2 mL), adjusted to pH ~ 8 with 1.0 M HCl aqueous solution, and then extracted with EtOAc (10 mL x 2). The organic layers were combined, washed with brine (15 mL), dried over Na2SO4, filtered and concentrated. The resulting residue was purified by flash silica gel chromatography ( 4g Purification on a silica gel flash column using a gradient of 0-15% ethyl acetate / petroleum ether at 15 mL / min afforded (3R,5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-17-(2-methyltetrahydrofuran-2-yl)-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol (11.2 mg, 11.4% yield) as a white solid. LCMS (ESI) m / z, C 24 H 40 O2: Calculated value 360.57, experimental value (M-OH) + :343.3. 1 H NMR (400MHz, CDCl3) δ (ppm) 4.05 (s, 1H) 3.92-3.88 (m, 1H) 3.79-3.77 (m, 1H) 2.07-2.03(m,1H)1.87-1.60(m,10H)1.54-0.95(m,19H)0.79-0.74(m,7H). 13C NMR (100MHz, CDCl3) δ (ppm) 84.97, 68.26, 66.64, 59.50, 56.69, 54.38, 42.93, 40.29, 39.16, 37.99, 36.09,35.91,35.05,32.19,31.93,29.03,28.58,26.59,25.17,23.77,22.98,20.66,13.45,11.20.

[0264] Example 9: (3R,5R,8R,9R,10S,13S,14S,17S)-17-[3-(cyclopropylmethyl)oxetane-3-yl]-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0265]

[0266] Preparation of 2-((3R,5R,8R,9R,10S,13S,14S)-3-hydroxy-3,13-dimethylhexahydro-17H-cyclopentanediol Ethyl 1,7-phenanthren-1,7-ylidene) acetate

[0267]

[0268] To a solution of (3R,5R,8R,9R,10S,13S,14S)-3-hydroxy-3,13-dimethyl-1,2,4,5,6,7,8,9,10,11,12,14,15,16-tetradecahydrocyclopenta[a]phenanthrene-17-one (2.00 g, 6.89 mmol, 1 eq) in THF (20 mL) and ethanol (20 mL) was added sodium ethoxide (1.5 M in EtOH, 45.9 mL, 10 eq) and ethyl 2-diethoxyphosphoryl acetate (15.4 g, 68.9 mmol, 13.7 mL, 10 eq). The resulting mixture was stirred at 85 °C for 16 h and then concentrated. The resulting residue was diluted with EtOAc (100 mL), washed with 1.0 M HCl in H2O (50 mL), saturated aqueous NaHCO3 (50 mL), brine (50 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 80g Purification on silica gel flash column, 0-20% ethyl acetate / petroleum ether gradient at 60 mL / min eluent afforded ethyl 2-((3R,5R,8R,9R,10S,13S,14S)-3-hydroxy-3,13-dimethylhexahydro-17H-cyclopenta[a]phenanthren-17-ylidene)acetate (2.40 g, 96.6% yield) as a white solid.

[0269] Preparation of 2-[(3R,5R,8R,9R,10S,13R,14S,17R)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8, 9,10,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]acetic acid ethyl ester

[0270]

[0271] To a solution of ethyl 2-((3R,5R,8R,9R,10S,13S,14S)-3-hydroxy-3,13-dimethylhexahydro-17H-cyclopenta[a]phenanthrene-17-ylidene)acetate (3.00 g, 8.32 mmol, 1 equiv) in EtOH (40 mL) under N2 atmosphere was added Pd / C (400 mg, 10 wt% loading), and the mixture was stirred under H2 (15 psi) at room temperature for 18 h. The resulting reaction mixture was filtered, and the filtrate was concentrated to give ethyl 2-[(3R,5R,8R,9R,10S,13R,14S,17R)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]acetate (3.00 g, 99.4% yield) as a colorless oil. 1 H NMR (400MHz, CDCl3) δ (ppm) 4.12 (q, J = 6.8Hz, 2H), 2.36 (dd, J = 4.8, 14.4Hz, 1H), 2.11 (dd, J = 9.6, 14.4Hz, 1H), 1.97-1.75(m,5H),1.67-1.60(m,3H),1.50-1.37(m,6H),1.36-1.20(m,11H),1.18-0.97(m,6H),0.60(s,3H).

[0272] Preparation of 2-[(3R,5R,8R,9R,10S,13S,14S,17R)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8, 9,10,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]malonic acid diethyl ester

[0273]

[0274] To a solution of DIPA (1.40 g, 13.8 mmol, 1.95 mL, 2.5 equiv) in THF (40 mL) was added n-BuLi (2.5 M in hexanes, 5.52 mL, 2.5 equiv) dropwise at 0 °C. The mixture was stirred at 0° C. for 0.5 h, and then ethyl 2-[(3R,5R,8R,9R,10S,13R,14S,17R)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetrahydro-1H-cyclopenta[a]phenanthren-17-yl]acetate (2.00 g, 5.52 mmol, 1 eq) and N-[bis(dimethylamino)phosphoryl]-N-methyl-methanamine (1.0 M, 5.52 mL, 1 eq) were added dropwise at −78° C. The resulting mixture was stirred at −78° C. for another 0.5 h, and then ethyl chloroformate (1.22 g, 11.3 mmol, 1.07 mL, 2.04 eq) was added dropwise at −78° C., and the mixture was then stirred at −78° C. for 4 h. The reaction mixture was quenched with saturated NH4Cl aqueous solution (50 mL) and extracted with EtOAc (50 mL x 2). The organic layers were combined, washed with salt water (50 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 80g Purification on a silica gel flash column with a gradient of 0-20% ethyl acetate / petroleum ether at 40 mL / min eluent afforded diethyl 2-[(3R,5R,8R,9R,10S,13S,14S,17R)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]malonate (1.35 g, 59% yield) as a yellow solid. LCMS (ESI) m / z, C 26 H 42 O5: Calculated 434.30, Found (M+Na) + :457.2. 1 H NMR (400MHz, CDCl3) δ (ppm) 4.22-4.11 (m, 4H), 3.30 (d, J = 11.6Hz, 1H), 2.26-2.16 (m, 1H), 2.00-1.88(m,1H),1.87-1.77(m,3H),1.65-1.61(m,2H),1.49-1.02(m,27H),0.71(s,3H).

[0275] Preparation of 2-(cyclopropylmethyl)-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl 2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]malonic acid Diethyl ester

[0276]

[0277] To a solution of diethyl 2-[(3R,5R,8R,9R,10S,13S,14S,17R)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]malonate (250 mg, 0.58 mmol, 1 eq) in DMF (5 mL) was added NaH (69 mg, 1.74 mmol, 60% in mineral oil, 3 eq) at 0 ° C. The resulting mixture was stirred at 0 ° C for 0.5 h, then iodomethylcyclopropane (209 mg, 1.15 mmol, 2 eq) was added, and the mixture was stirred at 50 ° C for another 16 h. The reaction mixture was then quenched with H O (5 mL) at 0 ° C. and extracted with EtOAc (5 mL x 3). The organic layers were combined, washed with brine (15 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 4g Purification by silica gel flash column (eluting with a 0-25% ethyl acetate / petroleum ether gradient at 20 mL / min) afforded diethyl 2-(cyclopropylmethyl)-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]malonate (170 mg, 60% yield) as a light yellow oil.

[0278] Preparation of 2-(cyclopropylmethyl)-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl 2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propane-1, 3-diol

[0279]

[0280] To a solution of diethyl 2-(cyclopropylmethyl)-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl]malonate (170 mg, 0.35 mmol, 1 eq) in THF (10 mL) was added LiAlH4 (66 mg, 1.75 mmol, 5 eq) at 0 ° C. The resulting mixture was stirred at 20 ° C for 16 h. The reaction mixture was quenched with aqueous NaOH (1.0 M, 3 mL) and extracted with EtOAc (5 mL x 3). The organic layers were combined, washed with brine (15 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 4g The product was purified by silica gel flash column (eluting with a gradient of 25-75% ethyl acetate in petroleum ether at 20 mL / min) to afford 2-(cyclopropylmethyl)-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propane-1,3-diol (120 mg, 85% yield) as a white solid.

[0281] Preparation of (3R,5R,8R,9R,10S,13S,14S,17S)-17-[3-(cyclopropylmethyl)oxetane-3-yl]- 3,13-Dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0282]

[0283] To a solution of 2-(cyclopropylmethyl)-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propane-1,3-diol (70 mg, 0.17 mmol, 1 eq) in DMF (3 mL) was added NaH (27.7 mg, 0.69 mmol, 60% in mineral oil, 4 eq). The resulting mixture was stirred at 20 °C for 0.5 h, then TsCl (42.3 mg, 0.22 mmol, 1.3 eq) was added, and the mixture was stirred at 20 °C for an additional 16 h. The reaction mixture was quenched with H2O (5 mL) at 0°C and extracted with EtOAc (5 mL x 3). The organic layers were combined, washed with brine (15 mL), dried over Na2SO4, filtered and concentrated. The resulting residue was purified by flash silica gel chromatography ( 4g The product was purified by silica gel flash column with a gradient of 0-20% ethyl acetate / petroleum ether at 20 mL / min to afford (3R,5R,8R,9R,10S,13S,14S,17S)-17-[3-(cyclopropylmethyl)oxetan-3-yl]-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol (19.7 mg, 29% yield) as a white solid. LCMS (ESI) m / z, C 26 H 42 O2: Calculated value 386.32, experimental value (M-OH) + :369.2. 1 H NMR (400MHz, CDCl3) δ (ppm) 4.88 (d, J = 6.02Hz, 1H), 4.61-4.56 (m, 1H), 4.55-4.5 1(m,1H),4.35(d,J=6.5Hz,1H),2.15-2.03(m,1H),1.99-1.89(m,2H),1.87-1.78 (m,4H),1.73-1.55(m,6H),1.50-1.34(m,6H),1.29-1.19(m,8H),1.12-0.99(m, 3H),0.96-0.88(m,1H),0.60-0.50(m,5H),0.26-0.17(m,1H),0.12-0.03(m,1H). 13 C NMR (100MHz, CDCl3) δ (ppm) 75.9,74.3,68.0,51.7,48.6,42.3,39.7,38.6,37.2,36 .3,36.1,33.6,30.7,30.5,27.4,22.5,22.0,21.6,21.4,20.6,20.2,8.4,2.4,1.4.

[0284] Example 10: (3R,5R,8R,9R,10S,13S,14S,17S)-17-(3-fluorooxetane-3-yl)-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0285]

[0286] Preparation of 2-fluoro-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6, 7,8,9,10,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]malonic acid diethyl ester

[0287]

[0288] To a solution of diethyl 2-[(3R,5R,8R,9R,10S,13S,14S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]malonate (500 mg, 1.15 mmol, 1 eq) in DMF (20 mL) was added NaH (230 mg, 5.75 mmol, 60% in mineral oil, 5 eq) at 0° C. The mixture was stirred at 0° C. for 0.5 h, and then 1-(chloromethyl)-4-fluoro-1,4-diazabicyclo[2.2.2]octane ditetrafluoroborate (1.22 g, 3.45 mmol, 3 eq) was added at 0° C. The resulting mixture was stirred at room temperature for 16 h. The reaction mixture is quenched by saturated NH4Cl aqueous solution (20mL), and extracted with EtOAc (15mL x 2). The organic layers are merged, washed with salt water (15mL), dried over Na2SO4, filtered and concentrated. The residue is purified by flash silica gel chromatography ( 12g Purification by silica gel flash column, 0-10% ethyl acetate / dichloromethane gradient at 20 mL / min eluent afforded diethyl 2-fluoro-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]malonate (470 mg, 90% yield) as a white solid. 1 H NMR (400MHz, CDCl3) δ (ppm) 4.38-4.17 (m, 4H), 2.56-2.38 (m, 1H), 1.87-1.75 (m, 5H), 1.66-1.55 (m, 7H ),1.44-1.37(m,4H),1.34-1.25(m,12H),1.20-1.12(m,3H),1.10-1.03(m,2H),0.81(d,J=4.8Hz,3H). 19 F NMR (376MHz, CD Cl3) δ (ppm)-174.29.

[0289] Preparation of 2-fluoro-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6, 7,8,9,10,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propane-1,3-diol

[0290]

[0291] To a solution of LiAlH4 (394 mg, 10.4 mmol, 10 equivalents) in THF (50 mL) was added diethyl 2-fluoro-2-[(3R, 5R, 8R, 9R, 10S, 13S, 14S, 17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetrahydro-1H-cyclopenta[a]phenanthrene-17-yl]malonate (470 mg, 1.04 mmol, 1 equivalent). The mixture was stirred at room temperature for 4 h. The reaction mixture was quenched with H2O (10 mL) and extracted with EtOAc (15 mL x 2). The organic layers were combined, washed with brine (10 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 12g Purification by silica gel flash column, 0-10% methanol / dichloromethane gradient at 20 mL / min eluent afforded 2-fluoro-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propane-1,3-diol (190 mg, 49% yield) as a white solid.

[0292] Preparation of (3R,5R,8R,9R,10S,13S,14S,17S)-17-(3-fluorooxetane-3-yl)-3,13-dimethyl 2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0293]

[0294] To a solution of 2-fluoro-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propane-1,3-diol (190 mg, 0.52 mmol, 1 eq) in DMF (8 mL) was added NaH (103 mg, 2.58 mmol, 60% in mineral oil, 5 eq) at 0° C. The mixture was stirred at 0° C. for 1 h, then 4-methylbenzenesulfonyl chloride (118 mg, 0.62 mmol, 1.2 eq) was added at 0° C., and the resulting mixture was stirred at room temperature for another 15 h. The reaction mixture is quenched by saturated NH4Cl aqueous solution (20mL), and extracted with EtOAc (20mL x 2). The organic layer is merged, washed with salt water (20mL), dried over Na2SO4, filtered and concentrated. The residue is purified by flash silica gel chromatography ( 4g Purification by silica gel flash column, 0-30% ethyl acetate / petroleum ether gradient at 20 mL / min eluent afforded (3R,5R,8R,9R,10S,13S,14S,17S)-17-(3-fluorooxetan-3-yl)-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol (65 mg, 36% yield) as a white solid. LCMS (ESI) m / z, C 22 H 35 FO2: Calculated 350.26, Exp. [M-OH] + :333.26. 1 H NMR (400MHz, CDCl3) δ (ppm) 4.89-4.60 (m, 4H), 2.00-1.79 (m, 7H), 1.74-1.63 (m, 2H), 1.48-1.36 (m, 5H), 1.33-0.98 (m, 14H), 0.65 (d, J = 1.2Hz, 3H). 19 F NMR (376MHz, CDCl3) δ (ppm)-147.27. 13 C NMR (100MHz, CDCl3) δ (ppm) 99.89,97.85,81.32,81.07,80.20,79.94,72.04,54.88,53.66, 43.58,41.17,40.35,39.28,37.73,34.74,31.39,26.47,26.06,25.42,24.25,22.87,12.87.

[0295] Example 11: (3R,5R,8R,9R,10S,13S,14S,17S)-17-[3-(hydroxymethyl)oxetan-3-yl]-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0296]

[0297] Preparation of 3-benzyloxy-2-cyano-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl Ethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propanoate ester

[0298]

[0299] To a solution of ethyl 2-cyano-2-[(3R,5R,8R,9R,10S,13S,14S,17R)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetrahydro-1H-cyclopenta[a]phenanthren-17-yl]acetate (4.00 g, 10.3 mmol, 1 eq) in THF (50 mL) was added LDA (2.5 M in THF, 10.3 mL, 2.5 eq) at -78 ° C under N2 atmosphere. The mixture was stirred at -78 ° C for 0.5 h, and then chloromethoxytoluene (2.40 g, 15.4 mmol, 2.1 mL, 1.5 eq) was added at -78 ° C. The resulting mixture was warmed to 20 ° C and stirred for another 16 h. The reaction mixture is quenched by saturated NH4Cl aqueous solution (15mL), and extracted with EtOAc (50mL x 2). The organic layer is merged, washed with salt water (40mL), dried over Na2SO4, filtered and concentrated. The residue is purified by flash silica gel chromatography ( 24g The mixture was purified by silica gel flash column (eluting with a gradient of 0-20% ethyl acetate in petroleum ether at 20 mL / min) to afford ethyl 3-benzyloxy-2-cyano-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propanoate (3.50 g, 66% yield) as a colorless oil.

[0300] Preparation of 2-(benzyloxymethyl)-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl 2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]-3-oxo- Ethyl propionate

[0301]

[0302] To a solution of ethyl 3-benzyloxy-2-cyano-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propanoate (2.00 g, 3.94 mmol, 1 eq) in DCM (30 mL) was added bis(cyclopentadienyl)zirconium chloride (4.26 g, 95% purity, 15.7 mmol, 4 eq). The resulting mixture was stirred at 20 ° C. under N 2 for 16 h. The reaction mixture was diluted with water (30 mL) and extracted with EtOAc (50 mL x 3). The organic layers were combined, washed with brine (50 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 20g The product was purified by silica gel flash column with a gradient of 0-20% ethyl acetate in petroleum ether at 30 mL / min to afford 2-(benzyloxymethyl)-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]-3-oxo-propionic acid ethyl ester (1.70 g, 84% yield) as a white solid. 1 H NMR (400MHz, CDCl3) δ (ppm) 10.44 (s, 1H), 7.34-7.22 (m, 5H), 4.51-4.48 (d, J = 12.4Hz, 1H), 4.37-4.34 (d, J = 12.4Hz, 1H), 4.3 0-4.21(m,2H),4.00-3.98(d,J=8.0Hz,1H),3.65-3.63(d,J=8.0Hz,1H),2.04-1.77(m,8H),1.43-0.96(m,22H),0.63(s,3H).

[0303] Preparation of 2-(benzyloxymethyl)-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl 2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propane-1, 3-diol

[0304]

[0305] To a solution of ethyl 2-(benzyloxymethyl)-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl]-3-oxo-propionate (1.70 g, 3.33 mmol, 1 eq) in THF (40 mL) was added LiAlH4 (379 mg, 9.99 mmol, 3 eq). The resulting mixture was stirred at 20 ° C for 2 h, then quenched with 10% aqueous NaOH solution (8 mL) and extracted with EtOAc (15 mL x 3). The organic layers were combined, washed with brine (10 mL), dried over Na2SO4, filtered and concentrated. The resulting residue was purified by flash silica gel chromatography ( 12g The mixture was purified by silica gel flash column (eluting with a 0-70% ethyl acetate / petroleum ether gradient at 20 mL / min) to afford 2-(benzyloxymethyl)-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propane-1,3-diol (1.30 g, 83% yield) as a colorless oil.

[0306] Preparation of (3R,5R,8R,9R,10S,13S,14S,17S)-17-[3-(benzyloxymethyl)oxetane-3-yl]- 3,13-Dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0307]

[0308] To a solution of 2-(benzyloxymethyl)-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propane-1,3-diol (1.20 g, 2.55 mmol, 1 eq) in DMF (6 mL) was added NaH (204 mg, 5.10 mmol, 60% in mineral oil, 2 eq). The resulting mixture was stirred at 20 °C for 1 h, then 4-methylbenzenesulfonyl chloride (534 mg, 2.80 mmol, 1.1 eq) was added, and the mixture was stirred at 20 °C for 2 h. The reaction mixture was quenched with water (10 mL) and extracted with EtOAc (15 mL x 3). The organic layers were combined, washed with salt water (10 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 12g The product was purified by silica gel flash column (eluting with a gradient of 0-40% ethyl acetate in petroleum ether at 20 mL / min) to afford (3R,5R,8R,9R,10S,13S,14S,17S)-17-[3-(benzyloxymethyl)oxetan-3-yl]-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol) as a white solid (650 mg, 56% yield). 1 H NMR (400MHz, CDCl3) δ (ppm) 7.37-7.30 (m, 4H), 4.85-4.83 (d, J = 6.4Hz, 1H), 4. 62-4.59(d,J=12.0Hz,1H),4.55-4.52(m,2H),4.46-4.44(d,J=5.6Hz,1H),4. 24-4.23(d,J=6.4Hz,1H),3.91-3.89(d,J=8.8Hz,1H),3.68-3.66(d,J=8.8Hz ,1H),2.12-2.09(m,1H),1.84-1.79(m,7H),1.44-1.00(m,20H),0.52(s,3H).

[0309] Preparation of (3R,5R,8R,9R,10S,13S,14S,17S)-17-[3-(hydroxymethyl)oxetane-3-yl]-3, 13-Dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0310]

[0311] To a solution of (3R, 5R, 8R, 9R, 10S, 13S, 14S, 17S) -17- [3- (benzyloxymethyl) oxetane-3-yl] -3, 13- dimethyl -2, 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 15, 16, 17- tetrahydro -1H- cyclopenta [a] phenanthren-3-ol (650 mg, 1.44 mmol, 1 equiv) in MeOH (2 mL) and THF (4 mL) was added Pd / C (50 mg, 10 wt% load) and Pd (OH) / C (50 mg, 20 wt% load). The mixture was stirred at 20 ° C under H (15 Psi) for 16 h. The resulting reaction mixture was filtered and the filtrate was concentrated. The residue was purified by flash silica gel chromatography ( 4g Purification on a silica gel flash column using a gradient of 0-80% ethyl acetate / petroleum ether at 20 mL / min afforded (3R,5R,8R,9R,10S,13S,14S,17S)-17-[3-(hydroxymethyl)oxetan-3-yl]-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol (360 mg, 69% yield) as a white solid. LCMS (ESI) m / z, C 23 H 38 O3: Calculated 362.28, Found [M-OH] + :345.3. 1 H NMR (400MHz, CDCl3) δ (ppm) 4.87-4.86 (d, J=6.8Hz, 1H), 4.56-4.55 (d, J=6.0Hz, 1H), 4.47-4.46 (d, J=5.6Hz, 1H), 4.2 5-4.24(d,J=6.4Hz,1H),4.11-4.10(m,1H),3.86-3.85(m,1H),2.14-1.68(m,10H),1.40-1.03(m,17H),0.54(s,3H). 13 C NMR(100MH z, CDCl3)δ(ppm)78.74,74.85,72.03,67.07,55.47,49.50,47.09,43.25,41.16,41.13,40 .29,39.51,37.53,34.65,34.47,31.37,26.42,25.93,25.52,25.36,24.18,24.09,12.31.

[0312] Example 12: 3-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]oxetane-3-carboxylic acid

[0313]

[0314] Preparation of 3-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8, 9,10,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]oxetane-3-carboxylic acid

[0315] To a solution of (3R,5R,8R,9R,10S,13S,14S,17S)-17-[3-(hydroxymethyl)oxetan-3-yl]-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol (30 mg, 0.083 mmol, 1 eq) in MeCN (1.5 mL) and HO (1 mL) were added TEMPO (2.6 mg, 0.017 mmol, 0.2 eq), NaClO (123 mg, 0.17 mmol, 10% in HO, 2 eq), sodium chlorite (35 mg, 0.33 mmol, 85%, 4 eq) and sodium dihydrogen phosphate dihydrate (51 mg, 0.33 mmol, 4 eq). The resulting mixture was stirred at 50°C for 16 h and then concentrated. The residue was purified by flash silica gel chromatography ( 4g Purification on a silica gel flash column using a 0-5% methanol / dichloromethane gradient at 15 mL / min afforded 3-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]oxetane-3-carboxylic acid (5.7 mg, 18% yield) as a white solid. LCMS (ESI) m / z, C 23 H 36 O4: Calculated 376.26, experimental [MO H] + :359.3. 1 H NMR (400MHz, CDCl3) δ (ppm) 5.05-5.03 (d, J = 6.8Hz, 1H) 4.82-4.81 (d, J = 6.4Hz, 1H) 4.70-4.64 (m, 2H) 2.06 (s, 3H) 1.80-0.85 (m, 24H) 0.54 (s, 3H). 13 C NMR (100MHz, CDCl3) δ (ppm) 177.97,79.00,74.15,72.82,55.05,52.15,50.01,43.27,41.20, 41.03,40.29,37.62,37.51,34.70,34.26,31.36,29.71,26.44,25.97,25.39,23.55,11.93.

[0316] Example 13: 2-[3-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]oxetane-3-yl]acetonitrile

[0317]

[0318] Preparation of ethyl[3-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6, 7,8,9,10,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]oxetane-3-yl]methane Methanesulfonate

[0319]

[0320] To a solution of (3R, 5R, 8R, 9R, 10S, 13S, 14S, 17S) -17- [3- (hydroxymethyl) oxetan-3-yl] -3, 13- dimethyl -2, 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 15, 16, 17- tetrahydro -1H- cyclopenta [a] phenanthren-3-ol (50 mg, 0.14 mmol, 1 eq) in DCM (2 mL) was added DMAP (50.5 mg, 0.41 mmol, 3 eq). The resulting mixture was stirred at 20 ° C for 0.5 h, then MsCl (18.9 mg, 0.16 mmol, 1.2 eq) was added, and the mixture was stirred at 20 ° C for another 16 h. The reaction mixture was quenched with water (2 mL) and extracted with EtOAc (5 mL x 2). The organic layers were combined, washed with brine (5 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 4g The product was purified by silica gel flash column (eluting with a gradient of 0-35% ethyl acetate in petroleum ether at 20 mL / min) to afford [3-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]oxetan-3-yl]methyl methanesulfonate (30 mg, 49% yield) as a white solid.

[0321] Preparation of 2-[3-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7, 8,9,10,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]oxetane-3-yl]acetonitrile

[0322]

[0323] To a solution of [3-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]oxetan-3-yl]methyl methanesulfonate (30 mg, 0.068 mmol, 1 eq) in DMF (1 mL) was added KCN (8.86 mg, 0.14 mmol, 2 eq). The mixture was stirred at 80 ° C. under N2 atmosphere for 16 h, then diluted with water (2 mL) and extracted with EtOAc (3 mL x 3). The aqueous phase was detoxified with 10% aqueous NaClO and adjusted to pH>11 with 1.0 M aqueous NaOH. The organic layers were combined, washed with brine (5 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 4g The product was purified by silica gel flash column with a gradient of 0-30% ethyl acetate / petroleum ether at 20 mL / min to afford 2-[3-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]oxetan-3-yl]acetonitrile as a white solid (8.9 mg, 33.8% yield). LCMS (ESI) m / z, C 24 H 37 NO2: Calculated value 371.28, experimental value [M-OH] + :354.28. 1 H NMR (400MHz, CDCl3) δ (ppm) 4.96-4.94 (d, J = 6.8Hz, 1H), 4.66-4.65 (d, J = 6.4Hz, 1H), 4.31-4.30 (d, J = 6.4Hz, 1H), 4.1 8-4.16(d,J=6.8Hz,1H),3.17-3.13(d,J=16.8Hz,1H),2.85-2.81(d,J=16.8Hz,1H),2.14-1.03(m,27H),0.54(s,3H). 13C NMR (100MHz, CDCl3) δ (ppm) 117.75, 80.25, 76.48, 71.96, 55.25, 51.31, 44.15, 43.52, 41.21, 41.17, 40.30,39.50,37.46,34.66,34.60,31.37,27.59,26.36,25.89,25.55,25.40,24.61,24.01,12.40.

[0324] Example 14: (3R,5R,8R,9R,10S,13S,14S,17S)-17-[3-(Fluoromethyl)oxetan-3-yl]-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0325]

[0326] Preparation of (3R,5R,8R,9R,10S,13S,14S,17S)-17-[3-(fluoromethyl)oxetane-3-yl]-3, 13-Dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0327] To a solution of [3-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl]oxetane-3-yl]methyl methanesulfonate (30 mg, 0.068 mmol, 1 eq) in THF (2 mL) was added TBAF (1.0 M in THF, 0.34 mL, 5 eq) at 25 °C. The resulting mixture was stirred at 60 °C for 16 h, then diluted with water (3 mL) and extracted with EtOAc (5 mL x2). The organic layers were combined, washed with brine (5 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 4g The product was purified by silica gel flash column with a gradient of 0-30% ethyl acetate / petroleum ether at 20 mL / min to afford (3R,5R,8R,9R,10S,13S,14S,17S)-17-[3-(fluoromethyl)oxetan-3-yl]-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol (8.8 mg, 35.5% yield) as a white solid. LCMS (ESI) m / z, C 23 H 37 FO2: Calculated 364.28, Exp. [M-OH]+ :347.0. 1 H NMR (400MH z, CDCl3) δ (ppm) 4.87-4.84 (m, 2H), 4.75-4.48 (m, 3H), 4.34-4.32 (d, J = 6.8Hz, 1H), 2.16-1.05 (m, 26H), 0.55 (s, 3H). 19 F NMR (376MHz, CDCl3) δ (ppm)-224.86. 13 C NMR(100MHz,CDCl3)δ(p pm)87.31,78.00,73.72,72.02,55.42,49.99,49.96,46.17,43.26,41.17,41.14,4 0.30,39.17,37.52,34.66,34.49,31.38,26.44,25.93,25.47,24.02,23.94,12.42.

[0328] Example 15: 1-[[3-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]oxetan-3-yl]methyl]pyrazole-4-carbonitrile

[0329]

[0330] Preparation of 1-[[3-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6, 7,8,9,10,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]oxetane-3-yl]methane 1,2-Dimethylpyrazole-4-carbonitrile

[0331] To a solution of [3-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]oxetane-3-yl]methyl methanesulfonate (10 mg, 0.023 mmol, 1 eq) and 1H-pyrazole-4-carbonitrile (2.11 mg, 0.023 mmol, 1 eq) in DMF (1 mL) was added KCO (9.41 mg, 0.068 mmol, 3 eq). The resulting mixture was stirred at 50 ° C for 16 h. It was then concentrated. The residue was purified by flash silica gel chromatography ( 4g The product was purified by silica gel flash column with a gradient of 0-30% ethyl acetate / petroleum ether at 20 mL / min to afford 1-[[3-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]oxetan-3-yl]methyl]pyrazole-4-carbonitrile (3.7 mg, 37.2% yield) as a white solid. LCMS (ESI) m / z, C 27 H 39 N3O2: Calculated 437.30, found (M+H) + :438.3. 1 H NMR (400MHz, CDCl3) δ (ppm) 7.89 (s, 1H), 7.84 (s, 1H), 4.98-4.96 (d, J = 6.8Hz, 1H), 4.65-4.62 (m, 2 H), 4.54-4.47 (m, 2H), 4.38 (d, J = 14.0Hz, 1H), 2.03-1.77 (m, 7H), 1.54-1.05 (m, 20H), 0.70 (s, 3H). 13 C NMR (100MHz, CDCl3) δ (ppm) 142.35, 135.45, 113.33, 92.38, 78.16, 75.24, 71.98, 58.28, 55.43, 50.89, 46.66, 43 .64,41.13,41.05,40.24,40.17,37.43,34.62,34.57,31.32,26.40,25.85,25.47,25.36,24.33,23.91,12.90.

[0332] Example 16 and Example 17: (3R,5R,8R,9R,10S,13S,14S,17S)-3,13-Dimethyl-17-[3-(triazol-2-ylmethyl)oxetane-3-yl]-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetrahydro-1H-cyclopenta[a]phenanthrene-3-ol and (3R,5R,8R,9R,10S,13S,14S,17S)-3,13-Dimethyl-17-[3-(triazol-1-ylmethyl)oxetane-3-yl]-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetrahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0333]

[0334] Preparation of (3R,5R,8R,9R,10S,13S,14S,17S)-3,13-dimethyl-17-[3-(triazol-2-ylmethyl)oxy] Heterocyclobutane-3-yl]-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene- 3-ol and (3R,5R,8R,9R,10S,13S,14S,17S)-3,13-dimethyl-17-[3-(triazol-1-ylmethyl)oxetane] [alkyl-3-yl]-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0335] To a solution of [3-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]oxetan-3-yl]methyl methanesulfonate (20 mg, 0.045 mmol, 1 eq) and 1,2,3-triazole (9.4 mg, 0.14 mmol, 3 eq) in DMF (1 mL) was added KCO (18.8 mg, 0.14 mmol, 3 eq). The resulting mixture was stirred at 60 ° C for 16 h and then concentrated. The residue was purified by flash silica gel chromatography ( 4g The product was purified by silica gel flash column with a gradient of 0-60% ethyl acetate / petroleum ether at 20 mL / min to give two products. (3R, 5R, 8R, 9R, 10S, 13S, 14S, 17S)-3,13-dimethyl-17-[3-(triazol-2-ylmethyl)oxetane-3-yl]-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol (4.0 mg, 14.2% yield) was obtained as a white solid. LCMS (ESI) m / z, C 25 H 39 N3O2: Calculated 413.30, found (M+H) + :414.3. 1 H NMR (400MHz, CDCl3) δ (ppm) 7.66 (s, 2H), 4.95-4.94 (d, J = 6.8Hz1H), 4.87 (d, J = 14 .0Hz,1H),4.68-4.61(m,4H),1.96-1.77(m,7H),1.42-1.02(m,20H),0.70(s,3H). 13 C NMR (100MHz, CDCl3) δ (ppm) 134.12,78.35,75.16,72.02,59.92,55.47,51.24,46.48,43.53,41.15, 41.10,40.27,39.81,37.47,34.67,34.52,31.36,26.46,25.91,25.47,25.37,24.25,23.98,12.77.

[0336] (3R,5R,8R,9R,10S,13S,14S,17S)-3,13-dimethyl-17-[3-(triazol-1-ylmethyl)oxetan-3-yl]-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol (5.6 mg, 23.8% yield) was obtained as a white solid. LCMS (ESI) m / z, C 25 H 39 N3O2: Calculated 413.30, found (M+H) + :414.3. 1 H NMR (400MHz, CDCl3) δ (ppm) 7.76 (s, 1H), 7.65 (s, 1H), 5.00-4.98 (d, J = 6.8Hz, 1H), 4.84 (d, J = 14.0 Hz,1H),4.67-4.63(m,2H),4.52-4.46(m,2H),2.04-1.76(m,6H),1.40-1.02(m,21H),0.71(s,3H). 13 C NMR (100MHz, CDCl3) δ (ppm) 133.60,124.61,78.15,75.32,71.96,55.88,55.39,50.89,46.55,43.65,41 .14,41.06,40.24,40.07,37.41,34.63,34.51,31.32,26.38,25.86,25.47,25.35,24.37,23.93,12.88.

[0337] Example 18: 3-((3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl)oxetane-3-carbonitrile

[0338]

[0339] Preparation of 3-((3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8, 9,10,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl)oxetane-3-carbaldehyde

[0340]

[0341] To a solution of (3R, 5R, 8R, 9R, 10S, 13S, 14S, 17S) -17- [3- (hydroxymethyl) oxetane -3- base] -3, 13- dimethyl -2, 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 15, 16, 17- tetrahydro -1H- cyclopenta [a] phenanthren-3-ol (50 mg, 0.14 mmol, 1 eq) in DCM (3 mL) was added Dess-Martin periodinane (118 mg, 0.28 mmol, 2 eq). The resulting mixture was stirred at 25 ° C for 2 h, then the mixture was diluted with saturated NaHCO (15 mL) and extracted with DCM (15 mL x 3). The organic layers were combined, washed with brine (20 mL), dried over Na2S O4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 4g The product was purified by silica gel flash column (0-50% ethyl acetate / petroleum ether gradient at 20 mL / min eluent) to afford 3-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]oxetane-3-carbaldehyde (30 mg, 60.3% yield) as a white solid.

[0342] Preparation of 3-((3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8, 9,10,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl)oxetane-3-carbaldehyde oxime

[0343]

[0344] To a solution of 3-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]oxetane-3-carbaldehyde (18 mg, 0.050 mmol, 1 eq) in EtOH (1 mL) was added pyridine (39 mg, 0.50 mmol, 10 eq) and hydroxylamine HCl (6.9 mg, 0.10 mmol, 2 eq). The mixture was stirred at 25 °C for 4 h and then concentrated. The residue was diluted with EtOAc (50 mL), washed with water (25 mL) and brine (25 mL), dried over Na2SO4, filtered and concentrated to give the crude product 3-((3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl)oxetane-3-carbaldehyde oxime (25 mg, crude) as a yellow solid which was used directly in the next step without further purification.

[0345] Preparation of 3-((3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8, 9,10,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl)oxetane-3-carbonitrile

[0346]

[0347] To a mixture of 3-((3R, 5R, 8R, 9R, 10S, 13S, 14S, 17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl)oxetane-3-carbaldehyde oxime (25 mg, 0.067 mmol, 1 equivalent) and CDI (43.2 mg, 0.27 mmol, 4 equivalents) in a microwave tube, THF (2 mL) was added. The resulting mixture was microwaved at 120 ° C for 20 min and then concentrated. The crude product was merged with the crude product from another batch and the combined crude product was purified by flash silica gel chromatography ( 4g The product was purified by silica gel flash column, 0-40% ethyl acetate / petroleum ether gradient at 20 mL / min eluent. The product was further purified by preparative TLC (SiO2, petroleum ether / ethyl acetate = 1 / 1) to give 3-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]oxetane-3-carbonitrile (4.8 mg, 20.2% yield) as a white solid. LCMS (ESI) m / z, C 23 H 35 NO2: Calculated value 357.27, experimental value [M-OH] + :340.26. 1 H NMR (400MHz, CDCl3) δ (ppm) 4.90 (dd, J = 8.0, 4.0Hz, 2H), 4.72 (dd, J = 12.0, 4.0Hz, 2H), 2.10-2.05 (m, 1H), 1.95-1.87 (m,3H),1.82-1.74(m,4H),1.71-1.62(m,4H),1.47-1.39(m,5H),1.36-1.26(m,7H),1.20-1.01(m,4H),0.76(s,3H). 13 C NMR (100MHz, CDCl3) δ (ppm) 122.21,78.94,77.66,72.03,54.69,53.97,43.89,41.10,41.08,40. 23,39.03,38.56,37.66,34.66,34.46,31.29,26.50,25.95,25.39,25.25,23.77,23.72,13.33.

[0348] Example 19: (3R,5R,8R,9R,10S,13S,14S,17S)-17-[3-(difluoromethyl)oxetan-3-yl]-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0349]

[0350] Preparation of 2-(difluoromethyl)-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl- 2,4,5,6,7,8,9,10,11,12,14,15,16,17-Tetrahydro-1H-cyclopenta[a]phenanthren-17-yl]malonic acid diethyl ester

[0351]

[0352] To a solution of diethyl 2-[(3R,5R,8R,9R,10S,13S,14S,17R)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]malonate (150 mg, 0.35 mmol, 1 eq) in CHCN (10 mL) was added t-BuOK (77.5 mg, 0.70 mmol, 2 eq) followed by (bromodifluoromethyl)trimethylsilane (140 mg, 0.70 mmol, 2 eq). The resulting mixture was stirred at 25 °C for another 16 h, then diluted with H0 (5 mL) and extracted with EtOAc (5 mL x 3). The organic layers were combined, washed with brine (15 mL), dried over Na2SO4, filtered and concentrated. The resulting residue was purified by flash silica gel chromatography ( 12g Purification by silica gel flash column (eluting with a 0-5% ethyl acetate / petroleum ether gradient at 20 mL / min) afforded diethyl 2-(difluoromethyl)-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]malonate (110 mg, 65% yield) as a light yellow oil.

[0353] Preparation of 2-(difluoromethyl)-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl- 2,4,5,6,7,8,9,10,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propane-1,3- diols

[0354]

[0355] At 0 ° C, to a solution of LiAlH4 (43.1 mg, 1.15 mmol, 5 equivalents) in THF (5 mL) was added a solution of 2- (difluoromethyl) -2- [(3R, 5R, 8R, 9R, 10S, 13S, 14S, 17S) -3-hydroxy -3,13- dimethyl -2,4,5,6,7,8,9,10,11,12,14,15,16,17- tetrahydro -1H- cyclopenta [a] phenanthrene -17- base] diethyl malonate (110 mg, 0.23 mmol, 1 equivalent). The resulting mixture was stirred at 25 ° C for 3h. The reaction mixture was then quenched with NaOH (1.0 M, 3 mL) and extracted with EtOAc (5 mL x 3). The organic layers were combined, washed with brine (15 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by preparative TLC (SiO 2 , petroleum ether / ethyl acetate = 1 / 1) to give 2-(difluoromethyl)-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propane-1,3-diol (20 mg, 27% yield) as a white solid. 1 H NMR (400MHz, CDCl3) δ (ppm) 6.18 (t, J = 56.0Hz, 1H), 4.03-3.94 (m, 4H), 2.11-2.05 (m, 2H), 1.98-1.77(m,8H),1.51-1.36(m,8H),1.31-1.27(m,6H),1.12-1.00(m,6H),0.80(s,3H).

[0356] Preparation of (3R,5R,8R,9R,10S,13S,14S,17S)-17-[3-(difluoromethyl)oxetane-3-yl]-3, 13-Dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0357]

[0358] To a solution of 2-(difluoromethyl)-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propane-1,3-diol (20 mg, 0.050 mmol, 1 eq) in DMF (5 mL) was added NaH (7.99 mg, 0.20 mmol, 60% in mineral oil, 4 eq) and TsCl (14 mg, 0.075 mmol, 1.5 eq) at 0° C. The resulting mixture was then stirred at 25° C. for 3 h. The reaction mixture was cooled to 0° C. and H O (5 mL) was added, followed by extraction with EtOAc (5 mL x 3). The organic layers were combined, washed with brine (10 mL), dried over Na2SO4, filtered and concentrated. The resulting residue was purified by preparative TLC (SiO2, petroleum ether / ethyl acetate = 3 / 1) to afford (3R, 5R, 8R, 9R, 10S, 13S, 14S, 17S) -17- [3- (difluoromethyl) oxetane-3-yl] -3, 13-dimethyl-2, 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 15, 16, 17-tetrahydro-1H-cyclopenta [a] phenanthren-3-ol (10.4 mg, 54% yield) as a white solid. LCMS (ESI) m / z, C 23 H 36 F2O2: calculated value 382.27, experimental value (M-OH) + :365.2. 1 H NMR (400MHz, CDCl3) δ (ppm) 6.06 (t, J = 56.0Hz, 1H), 4.81 (d, J = 6.8Hz, 1H), 4.73 (d, J = 6.4Hz, 1H), 4.54 (d, J = 7.2Hz, 1H), 4.48 (d, J = 6.0Hz, 1H), 2. 22-2.10(m,1H),2.08-1.97(m,1H),1.87-1.75(m,5H),1.70-1.59(m,4H ),1.52-1.35(m,6H),1.30-1.22(m,8H),1.15-1.00(m,3H),0.60(s,3H). 19 F NMR (376MHz, CDCl3) δ (ppm) -127.99, -128.14, -128.61, -128.94, -129.28, -129.75, -129.90. 13C NMR (100MHz, CDCl3) δ (ppm) 116.36,74.10,72.02,55.48,49.00,43.22,41.13,40. 31,39.25,37.51,34.64,34.52,31.38,26.45,25.92,25.47,25.38,23.96,12.84.

[0359] Example 20: (3R,5R,8R,9R,10S,13S,14S,17S)-3,13-dimethyl-17-(3-(2,2,2-trifluoroethyl)oxetane-3-yl)-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0360]

[0361] Preparation of 2-allyl-2-((3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4, 5,6,7,8,9,10,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl) diethyl malonate

[0362]

[0363] To a solution of diethyl 2-[(3R,5R,8R,9R,10S,13S,14S,17R)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]malonate (1.20 g, 2.76 mmol, 1 eq) in DMF (15 mL) was added NaH (552 mg, 13.8 mmol, 60% in mineral oil, 5 eq) at 0° C. The resulting mixture was stirred at 0° C. for 1 h, then 3-bromoprop-1-ene (501 mg, 4.14 mmol, 1.5 eq) was added at 0° C., and the mixture was stirred at 20° C. for another 16 h. The reaction mixture is quenched by saturated NH4Cl aqueous solution (30mL), and extracted with EtOAc (30mL x 2). The organic layers are merged, washed with salt water (10mL), dried over Na2SO4, filtered and concentrated. The residue is purified by flash silica gel chromatography ( 12g Purification on silica gel flash column, 0-20% ethyl acetate / petroleum ether gradient at 30 mL / min eluent afforded diethyl 2-allyl-2-((3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl)malonate (1.10 g, 83.9% yield) as a white solid. 1 H NMR (400MHz, CDCl3) δ (ppm) 5.90-5.73 (m, 1H), 5.09-4.95 (m, 2H), 4.23-4.04 (m, 4H), 2.99-2.88 (m, 1H), 2.63-2.52 ( m,1H),2.28-2.08(m,2H),1.94-1.76(m,5H),1.67-1.34(m,13H),1.28-1.23(m,8H),1.21-0.90(m,6H),0.67(s,3H).

[0364] Preparation of 2-allyl-2-((3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4, 5,6,7,8,9,10,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl)propane-1,3-diol

[0365]

[0366] To a solution of LiAlH4 (880 mg, 23.2 mmol, 10 equiv) in THF (120 mL) was added diethyl 2-allyl-2-[(3R, 5R, 8R, 9R, 10S, 13S, 14S, 17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetrahydro-1H-cyclopenta[a]phenanthrene-17-yl]malonate (1.10 g, 2.32 mmol, 1 equiv) at 0 ° C. The mixture was stirred at 20 ° C for 16 h. The reaction mixture was then quenched with aqueous NaOH (1.0 M, 4 mL) and H2O (3 mL), MgSO4 was added to the resulting mixture, and the mixture was stirred at 20 ° C for 0.5 h. The mixture was filtered and the filtrate was concentrated. The residue was purified by flash silica gel chromatography ( 12g The product was purified by silica gel flash column (0-75% ethyl acetate / petroleum ether gradient at 25 mL / min eluent) to afford 2-allyl-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propane-1,3-diol (680 mg, 75.1% yield) as a white solid.

[0367] Preparation of (3R,5R,8R,9R,10S,13S,14S,17S)-17-(3-allyloxetane-3-yl)-3,13- Dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0368]

[0369] To a solution of 2-allyl-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propane-1,3-diol (680 mg, 1.74 mmol, 1 eq) in DMF (12 mL) was added NaH (278 mg, 6.96 mmol, 60% in mineral oil, 4 eq) at 0° C. The resulting mixture was stirred at 0° C. for 0.5 h before the addition of 4-methylbenzenesulfonyl chloride (398 mg, 2.09 mmol, 1.2 eq) at 0° C. The mixture was stirred at 25 ° C for another 16 h, then saturated NH4Cl aqueous solution (20 mL) was added and extracted with EtOAc (30 mL x 2). The organic layers were combined, washed with brine (20 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 24g The product was purified by silica gel flash column (0-30% ethyl acetate / petroleum ether gradient at 25 mL / min eluent) to afford (3R,5R,8R,9R,10S,13S,14S,17S)-17-(3-allyloxetan-3-yl)-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol (470 mg, 1.26 mmol, 72.5% yield) as a white solid. 1H NMR (400MHz, CDCl3) δ (ppm) 6.03-5.86 (m, 1H), 5.24-5.12 (m, 2H), 4.90 (d, J = 6.4Hz, 1H), 4.49 (d, J = 5.6Hz, 1H), 4.31 (d, J = 5.6Hz, 1H), 4.23(d,J=6.0Hz,1H),2.76-2.63(m,1H),2.57-2.46(m,1H),2.14-2.01(m,1H),1.93-1.57(m,10H),1.47-1.02(m,17H),0.57(s,3H).

[0370] Preparation of 2-(3-((3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethylhexahydro-1H- cyclopenta[a]phenanthren-17-yl)oxetane-3-yl)acetaldehyde

[0371]

[0372] To a solution of (3R,5R,8R,9R,10S,13S,14S,17S)-17-(3-allyloxetane-3-yl)-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol (470 mg, 1.26 mmol, 1 eq) in THF (10 mL) and H2O (10 mL) was added potassium osmate dihydrate (46.5 mg, 0.13 mmol, 0.1 eq) and sodium periodate (809 mg, 3.78 mmol, 0.21 mL, 3 eq). The mixture was stirred at 20°C for 4 h, then quenched with saturated Na2S2O3 (10 mL) and extracted with EtOAc (10 mL x 2). The organic layers were combined, washed with brine (10 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 12g The product was purified by silica gel flash column (0-50% ethyl acetate / petroleum ether gradient at 25 mL / min eluent) to afford 2-[3-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]oxetan-3-yl]acetaldehyde (410 mg, 86.7% yield) as a white solid. 1H NMR (400MHz, CDCl3) δ (ppm) 9.90 (s, 1H), 4.97 (d, J = 6.4Hz, 1H), 4.71 (d, J = 6.4Hz, 1H), 4.40 (d, J = 6.4Hz, 1H), 4.25 (d, J = 6 .4Hz,1H),3.18-2.98(m,2H),2.18-2.05(m,1H),2.02-1.89(m,1H),1.75-1.55(m,6H),1.50-0.99(m,20H),0.55(s,3H).

[0373] Preparation of 2-(3-((3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7, 8,9,10,11,12,14,15,16,17-Tetrahydro-1H-cyclopenta[a]phenanthren-17-yl)oxetane-3-yl)acetic acid

[0374]

[0375] To a solution of 2-[3-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]oxetan-3-yl]acetaldehyde (71 mg, 0.19 mmol, 1 eq) in t-BuOH (2 mL), DCM (2 mL) and H2O (1 mL) was added 2-methyl-2-butene (52.4 mg, 0.75 mmol, 4 eq), NaClO2 (67.6 mg, 0.75 mmol, 4 eq) and NaH2PO4 (49.3 mg, 0.41 mmol, 2.2 eq). The resulting mixture was stirred at 25°C for 16 h. Reactant mixture is diluted with water (15mL), and extracted with EtOAc (15mL x 3).Organic layer is merged, washed with salt solution (20mL), through NaSODry, filter and concentrate.Crude product is merged with another batch, and grinds 10min with petroleum ether / EOAc (4mL, 1 / 1) at 25 DEG C.Mixture is filtered, and dry filter cake, obtain 2- [3- [(3R, 5R, 8R, 9R, 10S, 13S, 14S, 17S) -3- hydroxy -3,13- dimethyl -2,4,5,6,7,8,9,10,11,12,14,15,16,17- tetrahydro -1H- cyclopenta [a] phenanthrene -17- base] oxetanes -3- base] acetic acid (51mg, 68.5% yield) as white solid.

[0376] Preparation of (3R,5R,8R,9R,10S,13S,14S,17S)-3,13-dimethyl-17-(3-(2,2,2-trifluoroethyl) Oxetane-3-yl)-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a] Phenanthrene-3-ol

[0377]

[0378] To a solution of 2-[3-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]oxetan-3-yl]acetic acid (35 mg, 0.090 mmol, 1 equiv) in EtOAc (8 mL) was added 3,3-dimethyl-1-(trifluoromethyl)-1λ3,2-benzoiodooxolane (44.4 mg , 0.13mmol, 1.5 equivalents), 2-tert-butyl-1,1,3,3-tetramethylguanidine (7.8mg, 0.045mmol, 9.0uL, 0.5 equivalents), Ir[dF(CF3)ppy]2(dtbbpy)PF6(10mg), H2O(48mg, 2.69mmol, 30 equivalents), 3,4,7,8-tetramethyl-1,10-phenanthroline (6.4mg, 0.027mmol, 0.3 equivalents) and CuCl2(2.4mg, 0.018mmol, 0.2 equivalents). The reaction was stirred and irradiated at 25°C for 16h using a 40W blue LED lamp. The mixture was filtered and the filtrate was concentrated. The crude product was merged with another batch and the combined crude product was purified by preparative TLC (SiO2, DCM / EtOAc=3 / 1). The product was further purified by preparative HPLC (column: Boston Prime C18 150mm*30mm*5um; mobile phase: [water (0.225% FA)-ACN]; B%: 55%-85%, 9 min) to give (3R,5R,8R,9R,10S,13S,14S,17S)-3,13-dimethyl-17-[3-(2,2,2-trifluoroethyl)oxetan-3-yl]-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol (2.5 mg, 6.7% yield) as a white solid. 1H NMR (400MHz, CDCl3) δ (ppm) 4.95 (d, J = 8.0Hz, 1H), 4.64 (d, J = 8.0Hz, 1H), 4.55 (d, J = 8.0Hz, 1H), 4.35 (d, J = 8.0Hz, 1H), 2.87-2.74 (m, 1H), 2.68 -2.56(m,1H),2.08-1.92(m,2H),1.85-1.79(m,5H),1.68-1.64(m,5H) ,1.50-1.41(m,5H),1.32-1.22(m,8H),1.14-1.01(m,3H),0.65(s,3H). 19F NMR (376MHz, CDCl3) δ (ppm) -58.68.13C NMR (100MHz, CDCl3) δ (ppm) 125.27,78.98,77.93,72.03,55.51,51.49,43.87,42.97,41.19,41.14, 40.79,40.31,39.90,37.50,34.70,34.55,31.40,26.42,25.94,25.56,25.39,24.42,23.97,12.98.

[0379] Example 21: (3R,5R,8R,9R,10S,13S,14S,17S)-17-(3-cyclopropyloxetane-3-yl)-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0380]

[0381] Preparation of (3R,5R,8R,9R,10S,13S,14S)-3,13-dimethyl-17-methylene-1,2,4,5,6,7,8,9, 10,11,12,14,15,16-Tetradecahydrocyclopenta[a]phenanthrene-3-ol

[0382]

[0383] To a solution of methyltriphenylphosphonium bromide (3.69 g, 10.33 mmol, 3 equiv) in THF (15 mL) was added potassium tert-butoxide (1.16 g, 10.33 mmol, 3 equiv). The mixture was stirred at 60 ° C for 1 h, and then (3R, 5R, 8R, 9R, 10S, 13S, 14S) -3-hydroxy-3,13-dimethyl-1,2,4,5,6,7,8,9,10,11,12,14,15,16-tetradecahydrocyclopenta [a] phenanthrene-17-one (1.00 g, 3.44 mmol, 1 equiv) was added. The resulting mixture was stirred at 60 ° C for another 16 h, then diluted with EtOAc (50 mL), washed with brine (30 mL), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 12g Purification by silica gel flash column, 0-15% ethyl acetate / petroleum ether gradient at 25 mL / min eluent afforded (3R,5R,8R,9R,10S,13S,14S)-3,13-dimethyl-17-methylene-1,2,4,5,6,7,8,9,10,11,12,14,15,16-tetradecahydrocyclopenta[a]phenanthrene-3-ol (900 mg, 91% yield) as a white solid. 1H NMR (400MHz, CDCl3) δ (ppm) 4.63-4.61 (m, 2H), 2.58-2.41 (m, 1H), 2.31-2.16 (m, 1H), 1.94-1.78 ( m,4H),1.74-1.61(m,3H),1.50-1.39(m,5H),1.37-1.19(m,9H),1.17-1.05(m,4H),0.79(s,3H).

[0384] Preparation of (3R,5R,8R,9R,10S,13S,14S,17S)-17-(hydroxymethyl)-3,13-dimethyl-2,4,5,6,7, 8,9,10,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0385]

[0386] To a solution of (3R,5R,8R,9R,10S,13S,14S)-3,13-dimethyl-17-methylene-1,2,4,5,6,7,8,9,10,11,12,14,15,16-tetradecahydrocyclopenta[a]phenanthren-3-ol (900 mg, 3.12 mmol, 1 eq) in THF (20 mL) was added BH3·THF (1.0 M in THF, 9.4 mL, 3 eq). The resulting mixture was stirred at 25°C for 2 h, then aqueous NaOH (2.8 M, 9.4 mL, 8.4 eq) was slowly added at 0°C, followed by H2O2 (11.0 g, 97.0 mmol, 9.4 mL, 37% in H2O, 31 eq). The mixture was stirred at 25 ° C for another 16 h and extracted with EtOAc (100 mL x 2). The combined organic layers were washed with aqueous Na2S2O3 (10%, 100 mL) and brine (100 mL), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 12g Purification by silica gel flash column, 0-40% ethyl acetate / petroleum ether gradient at 30 mL / min eluent afforded (3R,5R,8R,9R,10S,13S,14S,17S)-17-(hydroxymethyl)-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol (900 mg, 94% yield) as a white solid. 1H NMR (400MHz, CDCl3) δ (ppm) 3.77-3.67 (m, 1H), 3.61-3.50 (m, 1H), 1.87-1.78 (m, 5H), 1. 67-1.61(m,5H),1.51-1.43(m,3H),1.34-1.24(m,10H),1.16-1.01(m,6H),0.66(s,3H).

[0387] Preparation of (3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9, 10,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthrene-17-carbaldehyde

[0388]

[0389] To a solution of (3R,5R,8R,9R,10S,13S,14S,17S)-17-(hydroxymethyl)-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol (900 mg, 2.94 mmol, 1 equiv) in DCM (40 mL) was added (1,1-diacetoxy-3-oxo-1λ 5 ,2-benzoiodine oxalyl-1-yl) acetate (2.49 g, 5.87 mmol, 2 equivalents). The mixture was stirred at 25 ° C for 3 h. The reaction mixture was then quenched with saturated aqueous NaHCO 3 solution (25 mL) and Na 2 S 2 O 3 (25 mL) and extracted with DCM (30 mL x 2). The organic layers were combined, washed with brine (50 mL), dried over anhydrous Na 2 SO 4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 25g Purification by silica gel flash column, 0-30% ethyl acetate / petroleum ether gradient at 30 mL / min eluent afforded (3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-17-carbaldehyde (480 mg, 54% yield) as a colorless oil. 1 H NMR (400MHz, CDCl3) δ (ppm) 9.77 (d, J = 2.0Hz, 1H), 2.36-2.26 (m, 1H), 2.14-2.08 (m, 1H), 2.02-1.96 (m,1H),1.90-1.68(m,10H),1.53-1.44(m,3H),1.30-1.20(m,9H),1.13-1.05(m,3H),0.75(s,3H).

[0390] Preparation of (3R,5R,8R,9R,10S,13S,14S,17S)-17-[cyclopropyl(hydroxy)methyl]-3,13-dimethyl- 2,4,5,6,7,8,9,10,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0391]

[0392] To a solution of (3R, 5R, 8R, 9R, 10S, 13S, 14S, 17S) -3-hydroxy-3, 13-dimethyl-2, 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 15, 16, 17-tetrahydro-1H-cyclopenta[a]phenanthrene-17-carbaldehyde (480 mg, 1.58 mmol, 1 equivalent) in THF (10 mL) was added bromo(cyclopropyl)magnesium (1.0 M in THF, 7.8 mL, 5 equivalents) at -78 ° C. The resulting mixture was stirred at 25 ° C for 16 h, then quenched with saturated NH4Cl aqueous solution (50 mL) and extracted with EtOAc (50 mL x 2). The combined organic layer was washed with brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 4g Purification by silica gel flash column, 0-20% ethyl acetate / petroleum ether gradient at 20 mL / min eluent afforded (3R,5R,8R,9R,10S,13S,14S,17S)-17-[cyclopropyl(hydroxy)methyl]-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol (450 mg, 82% yield) as a white solid.

[0393] Preparation of cyclopropyl-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6, 7,8,9,10,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]methanone

[0394]

[0395] To a solution of (3R,5R,8R,9R,10S,13S,14S,17S)-17-[cyclopropyl(hydroxy)methyl]-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol (450 mg, 1.30 mmol, 1 equiv) in DCM (25 mL) was added (1,1-diacetoxy-3-oxo-1λ 5,2-benzoiodine oxalyl-1-yl) acetate (1.10 g, 2.60 mmol, 2 equivalents). The resulting mixture was stirred at 25 ° C for 3 h, then quenched with saturated NaHCO3 aqueous solution (20 mL) and extracted with DCM (20 mL x 2). The combined organic layer was washed with brine (20 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 12g Purification on a silica gel flash column using a gradient of 0-20% ethyl acetate / petroleum ether at 20 mL / min afforded cyclopropyl-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]methanone (270 mg, 60% yield) as a white solid. LCMS (ESI) m / z, C 23 H 36 O2: Calculated value 344.27, found value (M+H) + :345.1. 1 H NMR (400Hz, CDCl3) δ (ppm) 2.78 (t, J = 8.8Hz, 1H), 2.29-2.17 (m, 1H), 2.15-2.08 (m, 1H), 1.94-1.77 (m, 4H), 1 .69-1.57(m,9H),1.57-1.48(m,2H),1.36-1.23(m,8H),1.12-1.05(m,3H),1.03-0.78(m,4H),0.59(s,3H). 13 CNMR(100Hz,CDCl3)δ(ppm)211.30,72.11,64.58,55.71,44.80,41.77,41.14,40.30,39.22,37. 68,34.73,34.48,31.37,26.49,26.08,25.73,25.44,24.31,22.20,21.37,13.74,11.14,10.70.

[0396] Preparation of (3R,5R,8R,9R,10S,13S,14S,17S)-17-(1-cyclopropylvinyl)-3,13-dimethyl-2, 4,5,6,7,8,9,10,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0397]

[0398] To a solution of cyclopropyl-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]methanone (140 mg, 0.41 mmol, 1 eq) in THF (10 mL) at -40 ° C was added trimethylsilylmethyllithium (0.56 M in pentane, 7.3 mL, 10 eq). The mixture was warmed to 25 ° C and stirred at 25 ° C for 16 h. The resulting mixture was then concentrated and the residue was diluted with MeOH (5 mL). 4-Methylbenzenesulfonic acid (706 mg, 4.10 mmol, 10 eq) was added to the resulting mixture, which was then stirred at 25 ° C for 1 h. The reaction mixture was diluted with saturated NaHCO3 aqueous solution (10 mL) and extracted with EtOAc (10 mL x 2). The combined organic layer was washed with salt water (10 mL), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 4g Purification by silica gel flash column, 0-10% ethyl acetate / petroleum ether gradient at 20 mL / min eluent afforded (3R,5R,8R,9R,10S,13S,14S,17S)-17-(1-cyclopropylvinyl)-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol (135 mg, 96% yield) as a colorless oil. LCMS (ESI) m / z, C 24 H 38 O: Calculated 342.29, Found [M-OH] + :325.2. 1 H NMR(400MHz, CDCl3)δ(ppm)4.63-4.61(m,2H),2.28-2.17(m,1H),1.98-1.92(m,1H),1.91-1.79(m,4 H),1.74-1.64(m,4H),1.48-1.41(m,4H),1.32-1.18(m,11H),1.11-0.99(m,3H),0.76-0.28(m,8H).

[0399] Preparation of (3R,5R,8R,9R,10S,13S,14S,17S)-17-(1-cyclopropyl-2-hydroxy-ethyl)-3,13-dimethyl 2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0400]

[0401] To a solution of (3R,5R,8R,9R,10S,13S,14S,17S)-17-(1-cyclopropylvinyl)-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol (135 mg, 0.40 mmol, 1 eq) in THF (15 mL) was added BH3·THF (1.0 M in THF, 4.0 mL, 10 eq). The mixture was stirred at 25°C for 2 h, then aqueous NaOH (2.8 M, 4.0 mL, 28 eq) was added dropwise at 0°C, followed by H2O2 (4.65 g, 41.0 mmol, 4.0 mL, 30% in H2O, 104 eq). The mixture was stirred at 25 ° C for another 16 h and extracted with EtOAc (20 mL x 2). The combined organic layers were washed with aqueous Na2S2O3 (10%, 20 mL), brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 4g The product was purified by silica gel flash column (eluting with a gradient of 0-50% ethyl acetate in petroleum ether at 20 mL / min) to afford (3R,5R,8R,9R,10S,13S,14S,17S)-17-(1-cyclopropylvinyl)-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol (140 mg, 97% yield) as a white solid.

[0402] Preparation of 2-cyclopropyl-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4, 5,6,7,8,9,10,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]acetaldehyde

[0403]

[0404] To a solution of (3R,5R,8R,9R,10S,13S,14S,17S)-17-(1-cyclopropyl-2-hydroxy-ethyl)-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol (140 mg, 0.39 mmol, 1 equiv) in DCM (10 mL) was added (1,1-diacetoxy-3-oxo-1λ 5,2-benzoiodine oxalyl-1-yl) acetate (330 mg, 0.78 mmol, 0.24 mL, 2 equivalents). The mixture was stirred at 25 ° C for 3 h, then quenched with saturated NaHCO 3 (10 mL) and extracted with EtOAc (20 mL x 2). The organic layers were combined, washed with brine (10 mL), dried over Na 2 SO 4, filtered and concentrated. The residue was purified by flash silica gel chromatography ( 4g Purification by silica gel flash column, 0-20% ethyl acetate / petroleum ether gradient at 20 mL / min eluent afforded 2-cyclopropyl-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]acetaldehyde (70 mg, 50% yield) as a colorless oil.

[0405] Preparation of 2-cyclopropyl-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4, 5,6,7,8,9,10,11,12,14,15,16,17-Tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propane-1,3-diol

[0406]

[0407] To a solution of 2-cyclopropyl-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]acetaldehyde (40 mg, 0.11 mmol, 1 eq) in EtOH (2 mL) and H2O (1.5 mL) was added aqueous NaOH (1.0 M, 0.56 mL, 5 eq) and HCHO (1.09 g, 13.4 mmol, 1.33 mL, 37% in H2O, 120 eq). The resulting mixture was stirred at 25° C. for 16 h. The reaction mixture is quenched by saturated NH4Cl aqueous solution (10mL), and extracted with EtOAc (10mL x 2). The organic layers are combined, washed with salt water (10mL), dried over Na2SO4, filtered and concentrated. The residue is purified by flash silica gel chromatography ( 4g Purification on a silica gel flash column with a 0-10% methanol / dichloromethane gradient at 20 mL / min eluent afforded 2-cyclopropyl-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propane-1,3-diol (15 mg, 23% yield, 66% purity) as a colorless oil. LCMS (ESI) m / z, C 25 H 42 O3: Calculated 390.31, Found [M-OH] + :373.3. 1 H NMR (400MHz, CDCl3) δ (ppm) 3.81-3.75 (m, 1H), 3.57-3.48 (m, 3H), 2.02-1.90 (m, 3H), 1.89-1.70 (m, 7H), 1.69-1.57 ( m,4H),1.53-1.43(m,2H),1.34-1.22(m,8H),1.21-0.97(m,6H),0.89(s,3H),0.86-0.78(m,1H),0.49-0.27(m,4H).

[0408] Preparation of (3R,5R,8R,9R,10S,13S,14S,17S)-17-(3-cyclopropyloxetane-3-yl)-3,13- Dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0409]

[0410] To a solution of 2-cyclopropyl-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]propane-1,3-diol (10 mg, 0.026 mmol, 1 eq) in DMF (2 mL) was added NaH (5.2 mg, 0.13 mmol, 60% in mineral oil, 5 eq) at 0° C. The mixture was stirred at 0° C. for 0.5 h, then 4-methylbenzenesulfonyl chloride (7.4 mg, 0.040 mmol, 1.5 eq) was added at 0° C. The mixture was stirred at 25 ° C for 16h, then quenched with saturated NH4Cl aqueous solution (10mL) and extracted with EtOAc (10mL x 2). The organic layers were combined, washed with brine (10mL), dried over Na2SO4, filtered and concentrated. The crude product was combined with another batch and purified by preparative TLC (SiO2, petroleum ether / ethyl acetate=3 / 1) to give (3R, 5R, 8R, 9R, 10S, 13S, 14S, 17S) -17- (3- cyclopropyloxetane -3- bases) -3,13- dimethyl -2,4,5,6,7,8,9,10,11,12,14,15,16,17- tetrahydro -1H- cyclopentadien [a] phenanthrene -3- alcohol (5.2mg, 19% yield). LCMS (ESI) m / z, C 25 H 40 O2: Calculated 372.30, Found [M-OH] + :355.2. 1 H NMR (400MHz, CDCl3) δ (ppm) 4.71 (d, J = 6.0Hz, 1H), 4.43 (d, J = 6.0Hz, 1H), 4.10-4.05 (m, 2H), 2.13-2.04 (m, 2H), 2.00-1.90 (m,1H),1.88-1.79(m,3H),1.76-1.65(m,2H),1.47-1.35(m,6H),1.30-1.22(m,9H),1.15-0.96(m,5H),0.74-0.41(m,8H). 13 C NMR (100MHz, CDCl3) δ (ppm) 78.99,75.26,72.08,56.17,55.54,44.86,43.53,41.26,41.19,40.36,39 .60,37.65,34.74,34.52,31.43,26.46,26.00,25.59,25.41,24.47,24.08,17.26,12.74,3.74,1.63.

[0411] Example 22: (3R,5R,8R,9R,10S,13S,14S,17S)-17-(3-ethynyloxetane-3-yl)-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0412]

[0413] Preparation of (3R,5R,8R,9R,10S,13S,14S,17S)-17-(3-ethynyloxetane-3-yl)-3,13- Dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0414] To a solution of 3-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl]oxetane-3-carbaldehyde (190 mg, 0.53 mmol, 1 eq) and KCO (145 mg, 1.05 mmol, 2 eq) in MeOH (6 mL) at 0°C was added 1-diazo-1-dimethoxyphosphoryl-propane-2-one (111 mg, 0.58 mmol, 1.1 eq). The resulting mixture was stirred at 20°C for 3 h, then filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography ( 24g Purification on a silica gel flash column using a gradient of 0-30% ethyl acetate / petroleum ether at 20 mL / min afforded (3R,5R,8R,9R,10S,13S,14S,17S)-17-(3-ethynyloxetan-3-yl)-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol (120 mg, 63.5% yield) as a white solid. LCMS (ESI) m / z, C 24 H 36 O2: Calculated 356.27, Found [M-OH] + :339.3. 1 HNMR (400MHz, CDCl3) δ (ppm) 4.74-4.70 (m, 4H), 2.46 (s, 1H), 2.04-1.07 (m, 27H), 0.71 (s, 3H). 13C NMR (100MHz, CDCl3) δ (ppm) 87.53, 81.95, 81.39, 73.63, 72.04, 55.10, 55.00, 44.03, 41.19, 40.35,39.59,39.28,37.74,34.77,34.53,31.41,26.46,26.05,25.43,23.84,23.69,13.35.

[0415] Example 23: (3R,5R,8R,9R,10S,13S,14S,17R)-17-(oxetan-3-yl)-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol

[0416]

[0417] (3R,5R,8R,9R,10S,13S,14S,17R)-17-(Oxetan-3-yl)-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetrahydro-1H-cyclopenta[a]phenanthrene-3-ol (12 mg, white solid) was prepared as described herein for (3R,5R,8R,9R,10S,13S,14S,17S)-17-[3-(cyclopropylmethyl)oxetan-3-yl]-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetrahydro-1H-cyclopenta[a]phenanthrene-3-ol , but using diethyl 2-(cyclopropylmethyl)-2-[(3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetrahydro-1H-cyclopenta[a]phenanthrene-17-yl]malonate instead of diethyl 2-[(3R,5R,8R,9R,10S,13S,14S,17R)-3-hydroxy-3,13-dimethyl-2,4,5,6,7,8,9,10,11,12,14,15,16,17-tetrahydro-1H-cyclopenta[a]phenanthrene-17-yl]malonate. LCMS (ESI) m / z, C 22 H 36 O2: Calculated 332.27, Found [M-OH] + :315.3. 1H NMR (400MHz, CDCl3) δ (ppm) 4.72-4.69 (m, 2H), 4.57-4.49 (m, 2H), 3.14-3.04 (m, 1H) 1.82-1.63 (m, 7H), 1.49-1.03 (m, 21H), 0.55 (s, 3H). 13 C NMR (100MHz, CDCl3) δ (ppm) 77.24, 72.03, 54.55, 54.41, 42.83, 41.59, 41.17, 40.36, 38. 76,37.86,36.69,34.79,34.52,31.40,26.43,26.14,25.47,25.35,25.18,24.29,13.06.

[0418] Determination method

[0419] The compounds provided herein can be evaluated in a variety of assays. A patch clamp electrophysiology assay is described here.

[0420] Cellular electrophysiology is used to measure the pharmacological properties of GABA receptor modulators. GABA A The channel was expressed in a stable cell line described herein. A The parental cell line of the channel is the human embryonic kidney (HEK293) cell line, which expresses the tetracycline repressor to support inducible expression of the target protein.

[0421] For GABA A In the α4β3δ cell line, the α4 and β3 subunits are under the control of a tetracycline-inducible expression system, while the δ subunit is constitutively expressed. A In the α1β2γ2 cell line, the α1 and β2 subunits are under the control of a tetracycline-inducible expression system, while the γ2 subunit is constitutively expressed.

[0422] Compounds targeting GABA were analyzed using the SyncroPatch automated platform in positive allosteric modulator (PAM) mode. A α1β2γ2 and GABA A Evaluation of α4β3δ. Six concentrations of 20 nM, 62 nM, 185 nM, 555 nM, 1667 nM, and 5000 nM were tested on the plate, with a single concentration of compound tested in each well. A minimum of three cells were harvested for each concentration of compound.

[0423] Automated patch-clamp recordings were performed using a SyncroPatch 384PE. Voltage protocols and data collection were performed using PatchController 384 V1.6.6 and Data Controller V1.6.0. Steady-state voltage pulses of -80 mV were applied during the measurement.

[0424] A stacking addition protocol was used in which γ-aminobutyric acid (GABA) was rapidly applied and then washed away from the cells. To test for positive allosteric modulator activity (PAM), the agonist GABA EC was first applied. 20 (EC 20 : 10 μM for GABA A α4β3δ channels and 14 μM GABA for GABA A α1β2γ2 channels) (with a wash step in between) were used twice as a control and to show reproducibility of activation, followed by preincubation of the test compound for 1-2 minutes and then application of GABA EC in the presence of the test compound. 20 Finally, after a further washing step, maximal GABA (10 mM) was applied.

[0425] Allopregnanolone concentration responses were tested on compound plates as a control PAM. Fold increases were generated using the following equation: (I comp / I 对照 )-1, where I comp is the current amplitude in the presence of the compound, and I 对照 In the presence of GABA EC alone 20 This establishes the current amplitude of the EC 50 Concentration-response curve, where 0 indicates no PAM activity and >0 indicates PAM activity.

[0426] The maximum % Emax for each compound was generated using the following equation: (I MaxComp / I AveMaxAllo )*100, where I MaxComp is the individual maximum increase in current for each compound, and I AveMaxAllo is the average maximum fold increase produced in the presence of allopregnanolone. max % values ​​were averaged. Compound and allopregnanolone values ​​were obtained from separate cells tested in the same compound run. Two GABA A Subtype.

[0427] The resulting EC50 values ​​and E max The % (relative to allopregnanolone) are summarized in Table 2.

[0428] Table 2. EC50 (μM) and E(subscript A) of compounds tested against GABAA receptors in PAM mode max (relative to allopregnanolone)

[0429]

[0430] EC 50 (nM) Range: A:EC 50 <500; B: 500 <EC 50 <1000; C:1000 <EC 50 <1500; D: 1500 <EC 50

[0431] E max % range (relative to allopregnanolone):+++:E max >90%;++:90%>E max >50%; +: E max <50%

Claims

1. A neuroactive steroid (NAS) having the following structure:

2. The neuroactive steroid according to claim 1, wherein the neuroactive steroid has the following structure:

3. The neuroactive steroid according to claim 1, wherein the neuroactive steroid has the following structure:

4. The neuroactive steroid according to claim 1, wherein the neuroactive steroid has the following structure:

5. The neuroactive steroid according to claim 1, wherein the neuroactive steroid has the following structure:

6. The neuroactive steroid according to claim 1, wherein the neuroactive steroid has the following structure:

7. A pharmaceutical composition comprising the neuroactive steroid (NAS) according to any one of claims 1 to 6; and a pharmaceutically acceptable excipient.

8. Use of a neuroactive steroid according to any one of claims 1 to 6 for the manufacture of a medicament for treating epilepsy in a subject.

9. Use of the neuroactive steroid of any one of claims 1 to 6 in the preparation of a medicament for treating major depressive disorder (MDD) in a subject.

10. Use of the neuroactive steroid according to any one of claims 1 to 6 in the preparation of a medicament for treating postpartum depression (PPD) in a subject.

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