NOVEL SUBSTITUTED AMIDES OF TRITERPENE DERIVATIVES AS HIV

INHIBITORS

This application claims the benefit of Indian provisional application no 4058/CHE/2015 filed on 05 ^th August 2015 which is hereby incorporated by reference in its entirety.

FIELD OF THE INVENTION

The present invention relates to novel substituted amides of triterpene derivatives and related compounds, compositions useful for therapeutic treatment of viral diseases and particularly HIV mediated diseases.

BACKGROUND OF THE INVENTION

The Human Immunodeficiency Virus (HIV) has now been established as the causative agent of the Acquired Immunodeficiency Syndrome (AIDS) for over 20 years (Science 1983, 220, 868-871; N.Eng.J.Med.1984, 311, 1292-1297). AIDS is characterized by the destruction of the immune system, particularly of CD4+T-cells. HIV is a retrovirus, and the HIV life cycle encompasses several crucial steps, starting from the attachment of the virus to the host cell membrane and finishing with the release of progeny virons from the cell.

The natural compound betulinic acid, isolated from Syzygium clavifolium and several other plant species was found to possess anti-HIV activity. Chemical modifications were undertaken by several research groups in an attempt to identify potent anti-HIV agents by making semi- synthetic analogs of betulinic acid, leading to the discovery of Bevirimat as a compound with a novel mechanism of action (J. Nat. Prod. 1994, 57(2): 243-7; J. Med. Chem. 1996, 39(5), 1016). Further studies shown that Bevirimat acts by disrupting Gag processing (Proc. Natl. Acad. Sci. USA 2003, 100 (23): 13555-60; Antimicrob. Agents. Chemother. 2001, 45(4), 1225-30; J. Virol. 2004, 78(2): 922-9; J. Biol. Chem. 2005, 280(51): 42149-55; J. Virol. 2006, 80(12): 5716-22) and to be a first-in-class maturation inhibitor with a potent activity against HIV-1. Bevirimat went up to phase 2 clinical trials, in clinic despite optimal plasma concentrations, not all patients given bevirimat have a robust viral load reduction. It was reported that non-respondant patients had more frequent base line Gag polymorphisms near the capsid SP-1 cleavage site than responders. (HIV gag polymorphism determines treatment response to bevirimat. XVII international HIV drug resistance work shop June 10-14, 2008, Sitges, Spain).

Encouraged by these developments, medicinal chemists started exploring betulinic acid derivatives and related compounds intensively for their therapeutic activities. For example, WO 2014/105926 describes novel betulinic acid proline derivatives as HIV inhibitors; WO 2013/160810 describes novel betulinic acid derivatives as HIV inhibitors; WO 2011/007230 describes lupeol-type triterpene derivatives as antivirals; WO 2011/153319 describes C-28 amides of modified C-3 betulinic acid derivatives as HIV maturation inhibitors; WO 2011/153315 describes modified C-3 betulinic acid derivatives as HIV maturation inhibitors; WO 2009/082819 describes novel lupane derivatives; WO 2009/100532 discloses novel 17 β lupine derivatives as anti-HIV agents; The patent publication WO 2014/093941 describers Pharmaceutical compositions of betulin derivatives; WO 2008/115281 describes preparation of triterpene derivatives for therapeutic use in the treatment of viral infections; WO 2007/002411 describes preparation of triterpenes as antiviral agents; WO 2006/053255 describes novel Betulin derivatives, Preparation thereof & use thereof; WO 2008/057420 extended triterpene derivatives as antiretroviral agents; US 2013/0096094 describes preparation of derivatives of 3-0-(3',3'-dimethylsuccinyl)-betulinic acid for the treatment of HIV; WO 2010/132334 describes 3,28-Disubstituted betulinic acid derivatives as Anti-HIV agents; US 2004/0204389 describes anti-HIV agents with dual sites of action.

Some additional references disclose betulinic acid related compounds. For example,

WO 2013/148067 describes preparation of 3,28-disubstituted betulinic acid derivatives as anti-HIV agents; WO 2012/154554 describes fatty acid triterpene derivatives and their uses; WO 2010/032123 describes preparation of triterpenoid compounds for pharmaceutical use; WO 2008/138200 describes preparation of lupane derivatives as NMDA and MC receptor antagonists exhibiting neuroprotective and memory enhancing activities; WO 2008/127364 describes preparation of betulinic acid derivatives for use in antiviral and anticancer pharmaceutical compositions; WO 2008/097341 describes antiviral compounds and use thereof; WO 94/26725 describes preparation of lupane antiviral agents; WO 2006/053255 describes novel Betulin derivatives, preparation thereof & use thereof; CN 103342729 describes caffeoyl-substituted pentacyclic triterpene derivative useful in treatment of various diseases and its preparation; CN 103127135 describes triterpene derivatives, their preparation method and use; CN 102108092 describes 23-Hydroxybetulinic acid derivative, its preparation method and application as antitumor agents and tumor multidrug resistance reversing agents; US 2011/0152229 describes betulinic acid derivatives as anti-HrV agents; CN 1861627 describes synthesis of 23-hydroxybetulinic acid derivatives and application as antitumor agents; EP 542622 describes preparation of [lup-20(29)-en-28- oylamino]alkanoates as antiviral agents; CN 101648988 describes preparation of pentacyclic triterpene-28-carboxylic amide derivative containing isoxazole ring and medical application; WO 2007/141392 describes compositions comprising betulonic acid; WO 2007/141391 describes betulin derived compounds useful as antiprotozoal agents; Bioorganic & Medicinal Chemistry Letters (2008), 18(24), 6377-6380, describes triterpene based compounds with potent anti-maturation activity against HIV-1; Bioorganic & Medicinal Chemistry (2006), 14(7), 2279-2289, describes synthesis and anti-HIV activity of bi-functional betulinic acid derivatives.

Given the fact of the world wide epidemic level of AIDS, there is a strong continued need for new effective drugs for treatment of HIV infected patients, disease conditions and/or disorders mediated by HIV by discovering new compounds with novel structures and/or mechanism of action(s).

SUMMARY OF THE INVENTION

The present invention relates to the com ounds of the formula (I):

wherein,

Ri can be substituted or unsubstituted alkyl, R ₂ can be hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted aminoacids, substituted or unsubstituted alkoxy or substituted or unsubstituted cycloalkyl;

X can be absent, O, S, CH ₂ or NR _a (wherein R _a is H, C(0)R _c , C(S)R _C , substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl; or R _a with its nitrogen and adjacent carbon together form N-contained heterocyclyl (Preferably, pyrrolidine, piperdine, piperazine, or morpholine));

Y can be C(O), C(S) or CR _e R _f (wherein R _e and R _f are independently selected from hydrogen, substituted or unsubstituted alkyl);

R ₃ and R ₄ can be independently selected from hydrogen, hydroxy, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, or substituted or unsubstituted aminoacids; wherein amino acids are substituted by substituted or unsubstituted alkyl, phosphoric acid, or phosphorus prodrugs; R ₃ and R ₄ can be taken together with their carbon atoms to which they are attached to form a bond; R ₃ and R4 can be taken together with their carbon atoms to which they are attached to form cycloalkyl; or R ₃ and R4 can be taken together with their carbon atoms to which they are attached to form epoxide;

R5 and R _c can be independently selected from hydrogen, C0 ₂ R _d (wherein R _d is hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl), or substituted or unsubstituted alkyl;

R ₆ , R7 and R ₈ can be independently selected from hydrogen, substituted or unsubstituted alkyl or substituted or unsubstituted cycloalkyl;

R ₇ and R ₈ can be taken together with carbon atom to which they are attached to form substituted or unsubstituted 3-7 membered cycloalkyl or heterocyclyl; be 4-7 membered heterocyclyl or 4-7 membered heteroaryl;

Zi, Z ₂ and Z ₃ can be independently selected from hydrogen, hydroxy, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted amino alkyl, or substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl; wherein the substituents are alkyl, cycloalkyl, alkoxy, halo, amino or haloalkyl.

Pharmaceutically acceptable salts of the compounds of the formula (I) are also contemplated. Likewise, pharmaceutically acceptable solvates, including hydrates, of the compounds of the formula (I) are contemplated.

It should be understood that the formula (I) structurally encompasses all stereoisomers, including enantiomers, diastereomers, racemates, and combinations thereof which may be contemplated from the chemical structure of the genus described herein. It should be understood that the formula (I) structurally encompasses all tautomers. Also contemplated are prodrugs of the compounds of the formula (I), including ester prodrugs.

According to one embodiment, there is provided a compound of formula (I), wherein

According to other embodiment, there is provided a compound of formula (I), wherein 'X' is absent.

According to yet another embodiment, there is provided a compound of formula (I), wherein R ₂ is hydrogen.

According to yet another embodiment, there is provided a compound of formula (I), when "X is absent, and R ₂ and R5 are hydrogen"; then R ₃ and R4 are taken together with carbon atoms to which they are attached to form a bond.

According to yet another embodiment, there is provided a compound of formula (I), when "X is absent, and R ₂ and R5 are hydrogen"; then R ₃ and R4 are taken together with carbon atoms to which they are attached form cycloalkyl (preferably cyclopropyl).

According to yet another embodiment, there is provided a compound of formula (I), wherein R ₆ is hydrogen.

According to yet another embodiment, there is provided a compound of formula (I), wherein R ₇ and R ₈ are hydrogen.

According to yet another embodiment, there is provided a compound of formula (I), wherein R ₇ is hydrogen and R ₈ is alkyl (preferably methyl or isopropyl).

According to yet another embodiment, there is provided a compound of formula (I), wherein R ₇ and R ₈ are alkyl (preferably methyl).

According to yet another embodiment, there is provided a compound of formula (I), wherein R ₇ and R ₈ can be taken together with carbon atom to which they are attached to form substituted or unsubstituted 3-7 membered cycloalkyl (preferably cyclopropyl, cyclobutyl or cyclopentyl).

Accordin to yet another embodiment, there is provided a compound of formula (I),

wherein

Accordingly, another aspect of the present invention provides compounds of formula

(IA): Formula (IA) , Ri, R ₂ , R ₃ , R ₄ , R5 X', R ₇ , Rg, i, Z ₂ and Z ₃ are as defined above including pharmaceutically acceptable salts, pharmaceutically acceptable solvates, pharmaceutically acceptable hydrates, tautomers, stereoisomers, prodrugs, or combination thereof.

In a further embodiment, there is provided a method for treating mammals infected with a virus, especially wherein said virus is HIV, comprising administering to said mammal an antiviral effective amount of a compound which is selected from the group of compounds of formula (I), and one or more pharmaceutically acceptable carriers, excipients or diluents. Optionally, the compound of formula (I) can be administered in combination with an antiviral effective amount of another AIDS treatment agent selected from the group consisting of: (a) an AIDS antiviral agent or (b) an anti-infective agent

Another embodiment of the present invention is a pharmaceutical composition comprising one or more compounds of formula (I), and one or more pharmaceutically acceptable carriers, excipients, and/or diluents; and optionally in combination with another AIDS treatment agent selected from the group consisting of: AIDS antiviral agent or anti- infective agent

In another embodiment of the present invention there is provided one or more methods for preparation of the compounds of formula (I).

In one further embodiment, the present invention also encompasses the method(s) of preparation of intermediates used in the preparation of compound of formula (I).

Below are the representative examples according to formula (I) , which are illustrative in nature only and are not intended to limit to the scope of the invention (Nomenclature has been generated from ChemBioDraw Ultra 13.0 version):

(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-

5a,5b, 8,8,1 la-pentamethyl-3a-(((S)- l-(5-phenyl- lH-imidazol-2-yl)ethyl)carbamo yl)-l-(prop- l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbon yl)cyclobutane-l-carboxylic acid (Example 1), 2.2- dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR,7aR,9S, 1 laR, 1 IbR, 13aR, 13bR)- 5a,5b,8,8, l la-pentamethyl-3a-(((S)-l-(5-phenyl-lH-imidazol-2-yl)ethyl)c arbamoyl)-l-(prop- l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)butano ic acid (Example 2),

3.3- dimethyl-5-oxo-5-(((lR,3aS,5aR,5bR,7aR,9S, 1 laR, 1 IbR, 13aR, 13bR)- 5a,5b,8,8, l la-pentamethyl-3a-(((S)-l-(5-phenyl-lH-imidazol-2-yl)ethyl)c arbamoyl)-l-(prop- l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)pentan oic acid (Example 3),

(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR, l lbR, 13aR, 13bR)-3a-(((S)-l-(l-(2- (dimethylamino)ethyl)-4-phenyl-lH-imidazol-2-yl)ethyl)carbam oyl)-5a,5b, 8,8, 1 la- pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chr ysen-9-yl)oxy)carbonyl)-2,2- dimethylcyclobutane-l-carboxylic acid (Example 4),

(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S, l laR,l lbR,13aR,13bR)- 5a,5b, 8,8, 1 la-pentamethyl-3a-((2-(5-phenyl- lH-imidazol-2-yl)propan-2-yl)carbamoyl)- 1- (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy) carbonyl)cyclobutane-l- carboxylic acid (Example 5),

2,2-dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR,7aR,9S, 1 laR, 1 IbR, 13aR, 13bR)- 5a,5b, 8,8, 1 la-pentamethyl-3a-((2-(5-phenyl- lH-imidazol-2-yl)propan-2-yl)carbamoyl)- 1- (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy) butanoic acid (Example 6),

(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((2-(l-(2- (dimethylamino)ethyl)-4-phenyl-lH-imidazol-2-yl)propan-2-yl) carbamoyl)-5a,5b, 8,8,1 la- pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chr ysen-9-yl) oxy)carbonyl)-2,2- dimethylcyclobutane-l-carboxylic acid (Example 7),

4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((2-(l-(2- (dimethylamino)ethyl)-4-phenyl-lH-imidazol-2-yl)propan-2-yl) carbamoyl)-5a,5b,8,8,l la- pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chr ysen-9-yl)oxy)-2,2-dimethyl- 4-oxobutanoic acid (Example 8),

(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 5a,5b,8,8,l la-pentamethyl-3a-((2-(4-phenyl-l-(2-(pyrrolidin-l-yl)ethyl) -lH-imidazol-2- yl)propan-2-yl)carbamoyl)-l-(prop-l-en-2-yl)icosahydro-lH-cy clopenta[a]chrysen-9- yl)oxy)carbonyl)cyclobutane-l-carboxylic acid (Example 9),

(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S, l laR,l lbR,13aR,13bR)- 5a,5b,8,8, l la-pentamethyl-3a-(((S)-2-methyl-l-(5-phenyl-lH-imidazol-2-y l)propyl) carbamoyl)- 1 -(prop- l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbon yl) cyclobutane-l-carboxylic acid (Example 10),

2,2-dimethyl-4-oxo-4-((( lR,3aS ,5aR,5bR,7aR,9S , 11 aR, 1 IbR, 13aR, 13bR)- 5a,5b,8,8,l la-pentamethyl-3a-(((S)-2-methyl-l-(5-phenyl-lH-imidazol-2- yl)propyl)carbamoyl)-l-(prop-l-en-2-yl)icosahydro-lH-cyclope nta[a]chrysen-9- yl)oxy)butanoic acid (Example 11),

(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclopropyl)-3a-((2-(5-phenyl-lH-i midazol-2- yl)propan-2-yl)carbamoyl)icosahydro-lH-cyclopenta[a]chrysen- 9-yl)oxy)

carbonyl)cyclobutane-l-carboxylic acid (Example 12),

(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 5a,5b, 8,8,1 la-pentamethyl-3a-((l -(5-phenyl- lH-imidazol-2-yl)cyclobutyl)carbamo yl)- 1- (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy) carbonyl)cyclobutane-l- carboxylic acid (Example 13),

2,2-dimethyl-4-oxo-4-((( lR,3aS ,5aR,5bR,7aR,9S , 11 aR, 1 IbR, 13aR, 13bR)- 5a,5b, 8, 8, l la-pentamethyl-3a-((l -(5-phenyl- lH-imidazol-2-yl)cyclobutyl)carbamoyl)-l- (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy) butanoic acid (Example 14),

(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(5-(4- fluorophenyl)-lH-imidazol-2-yl)cyclobutyl)carbamoyl)-5a,5b, 8,8,1 la-pentamethyl- l-(prop- l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbon yl)-2,2-dimethylcyclobutane- 1-carboxylic acid (Example 15),

4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(5-(4-fluorophenyl)-lH imidazol-2-yl)cyclobutyl)carbamoyl)-5a,5b,8,8,l la-pentamethyl- 1 -(prop- l-en-2- yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)-2,2-dimethyl -4-oxobutanoicacid (Exampl 16),

4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(5-(4-fluorophenyl)-lH imidazol-2-yl)cyclobutyl)carbamoyl)-5a,5b,8,8,l la-pentamethyl- 1-(1- methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)ox y)-2,2-dimethyl-4-oxo butanoic acid (Example 17),

2,2-dimethyl-4-oxo-4-((( lR,3aS ,5aR,5bR,7aR,9S , 11 aR, 1 IbR, 13aR, 13bR)- 5a,5b, 8, 8, l la-pentamethyl-3a-((l -(5-phenyl- lH-imidazol-2-yl)cyclopentyl)carbamoyl)-l- (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy) butanoic acid (Example 18),

(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 5a,5b, 8 ,8 , 11 a-pentamethyl-3 a-(( 1 -(5-phenyl- 1 H-imidazol-2-yl)cyclopropyl)carbamoyl) - 1 - (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy) carbonyl)cyclobutane-l- carboxylic acid (Example 19),

2,2-dimethyl-4-oxo-4-((( lR,3aS ,5aR,5bR,7aR,9S , 11 aR, 1 IbR, 13aR, 13bR)- 5a,5b, 8 ,8 , 11 a-pentamethyl-3 a-(( 1 -(5-phenyl- 1 H-imidazol-2-yl)cyclopropyl)carbamoyl) - 1 - (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy) butanoic acid (Example 20), (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(l-(2-

(dimethylamino)ethyl)-4-phenyl-lH-imidazol-2-yl)cycloprop yl)carbamoyl)-5a,5b,8,8,l la- pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chr ysen-9-yl)oxy)carbonyl)-2,2- dimethylcyclobutane-l-carboxylic acid (Example 21),

(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-

5a,5b, 8,8,1 la-pentamethyl-3a-(((5-phenyl- lH-imidazol-2-yl)methyl)carbamo yl)-l-(prop- 1- en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl )cyclobutane-l-carboxylic acid (Example 22), and

2,2-dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR,7aR,9S, 1 laR, 1 IbR, 13aR, 13bR)- 5a,5b, 8,8, 1 la-pentamethyl-3a-(((5-phenyl-lH-imidazol-2-yl)methyl)carbam oyl)-l-(prop-l- en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)butanoic acid (Example 23);

or pharmaceutically acceptable salts, pharmaceutically acceptable solvates, pharmaceutically acceptable hydrates, tautomers, stereoisomers, prodrugs are also contemplated.

The present invention also provides a pharmaceutical composition that includes at least one compound as described herein and at least one pharmaceutically acceptable excipient (such as a pharmaceutically acceptable carrier or diluent). Specifically, the pharmaceutical composition comprises a therapeutically effective amount of at least one compound described herein. The compound(s) present in the composition may be associated with a pharmaceutically acceptable excipient (such as a carrier or a diluent) or may be diluted by a carrier, or enclosed within a carrier which may be in the form of a capsule, sachet, or other container.

The compounds and pharmaceutical compositions described herein are useful in the treatment of diseases, conditions and/or disorders mediated by viral infections.

The present invention further provides a method of treating a disease, condition and/or disorder mediated by viral infections in a subject in need thereof by administering to the subject one or more compounds described herein in a therapeutically effective amount to cure that infection, specifically in the form of a pharmaceutical composition.

The invention provides a method for preventing; ameliorating or treating a HIV mediated disease, disorder or syndrome in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a compound of the invention. The invention further provides a method, wherein the HIV mediated disease, disorder or syndrome is like AIDS, AIDS related complex, or a syndrome characterized by symptoms such as persistent generalized lymphadenopathy, fever and weight loss, or a retroviral infection genetically related to AIDS. Anti HIV inhibitory potential of the compounds of present invention may be demonstrated by any one or more methodologies known in the art, such as by using the assays described in Mossman T, December 1983, Journal of immunological methods, 65 (1- 2), 55-63 and SPC Cole, cancer chemotherapy and Pharmacology, 1986, 17, 259-263.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides substituted amides of triterpene derivatives and related compounds, which may be used as antiviral particularly as anti-HIV compounds and processes for the synthesis of these compounds. Pharmaceutically acceptable salts, pharmaceutically acceptable solvates, enantiomers, diastereomers of the derivatives, together with pharmaceutically acceptable carriers, excipients or diluents, which can be used for the treatment of diseases, condition and/or disorders mediated by viral infections, are also provided.

The following definitions apply to the terms as used herein:

The terms "halogen" or "halo" includes fluorine, chlorine, bromine, or iodine.

The term "alkyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation, having from one to eight carbon atoms, and which is attached to the rest of the molecule by a single bond, e.g., methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), isobutyl, n-butyl, n-pentyl, and 1,1-dimethylethyl (t-butyl).

The term "amino" refers to -N¾.

The term "alkoxy" refers to a straight or branched hydrocarbon chain with oxygen radical consisting carbon and hydrogen atoms, containing saturation or unsaturation, having from one to eight carbon atoms, and which is attached through oxygen atom to the rest of the molecule by a single bond, e.g., methyloxy, ethyloxy, n-propyloxy, 1-methylethyloxy (isopropyloxy), n-butyloxy, n-pentyloxy, and 1 , 1 -dimethylethyloxy (t-butyloxy).

The term "cycloalkyl" denotes a non-aromatic mono or multicyclic ring system of 3 to about 12 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Examples of multicyclic cycloalkyl groups include, but are not limited to, perhydronapththyl, adamantyl and norbornyl groups, bridged cyclic groups and spirobicyclic groups, e.g., spiro (4,4) non-2-yl.

The term "amino acid(s)" refers to a straight or branched hydrocarbon chain containing an amine group, a carboxylic acid group, and a side-chain that is specific to each amino acid and which is attached through the nitrogen atom of the amine group to the rest of the molecule by a single bond, e.g., alanine, valine, isoleucine, leucine, phenylalanine, or tyrosine. The term "haloalkyl" refers to alkyl group (as defined above) is substituted with one or more halogens. A monohaloalkyl radical, for example, may have a chlorine, bromine, iodine or fluorine atom. Dihalo and polyhaloalkyl radicals may have two or more of the same or different halogen atoms. Examples of haloalkyl include, but are not limited to, chloromethyl, dichloromethyl, trichloromethyl, dichloroethyl, dichloropropyl, fluoromethyl, difluoromethyl, trifluoromethyl, pentafluoroethyl, heptafluoropropyl, difluoro chloromethyl, dichloro fluoromethyl, difluoroethyl, difluoropropyl and the like.

The term "aryl" refers to an aromatic radical having from 6 to 14 carbon atoms such as phenyl, naphthyl, tetrahydronapthyl, indanyl, and biphenyl.

The term 'hydroxy' refers to -OH group.

The terms "heterocyclyl" and "heterocyclic ring" refer to a saturated 3- to 15- membered ring radical which consists of carbon atoms and from one to five heteroatoms selected from nitrogen, phosphorus, oxygen and sulfur. For purposes of this invention, the heterocyclic ring radical may be a monocyclic, bicyclic or tricyclic ring system, which may include fused, bridged or spiro ring systems, and the nitrogen, phosphorus, carbon, oxygen or sulfur atoms in the heterocyclic ring radical may be optionally oxidized to various oxidation states. In addition, the nitrogen atom may be optionally quaternized; Examples of such heterocyclic ring radicals include, but are not limited to, , tetrahydroisouinolyl, piperidinyl, piperazinyl, azapanyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxoazepinyl, azepinyl, pyrrolyl, 4-piperidonyl, pyrrolidinyl, pyrazinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, decahydroisoquinolyl, tetrahydrofurtyl, tetrahydropyranyl, thienyl, benzothienyl, thiamorpholinyl, thiamorpholinyl sulfoxide, thiamorpholinyl sulfone, dioxaphospholanyl, oxadiazolyl. The heterocyclic ring radical may be attached to the main structure at any heteroatom or carbon atom that results in the creation of a stable structure.

The term "heteroaryl" refers to an aromatic heterocyclic ring radical. The heteroaryl ring radical may be attached to the main structure at any heteroatom or carbon atom that results in the creation of a stable structure. Examples of such heteroaryl ring include, but are not limited to imidazolyl, pyrazolyl, pyrrolyl, lH-pyrrolo[2,3-b]pyridinyl, tetrazoyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolinyl, oxazolidinyl, triazolyl, isoxazolyl, isoxasolidinyl, thiazolyl, thiazolinyl, thiazolidinyl, isothiazolyl, quinuclidinyl, isothiazolidinyl, indolyl, isoindolyl, indolinyl, isoindolinyl, quinolyl, isoquinolyl, benzimidazolyl, thiadiazolyl, benzothiazolyl, benzooxazolyl, furyl,

"Substituted" refers to 1-3 substituents on the same position or on different positions with the same groups or different groups. Unless otherwise specified, the term "substituted" as used herein refers to substitution with any one or any combination of the following substituents: hydroxy, halogen, carboxyl, cyano, nitro, oxo (=0), thio (=S), substituted or unsubstituted alkyl, haloalkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted amino, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocyclylalkyl ring, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted heterocyclic ring.

The term "prodrug" denotes a derivative of a compound, which derivative, when administered to warm -blooded animals, e.g. humans, is converted into the compound (drug). The enzymatic and/or chemical hydrolytic cleavage of the compounds of the present invention occurs in such a manner that the proven drug form (parent carboxylic acid drug) is released, and the moiety or moieties split off remain nontoxic or are metabolized so that nontoxic metabolic products are produced. For example, a carboxylic acid group can be esterified, e.g., with a methyl group or ethyl group to yield an ester. When an ester is administered to a subject, the ester is cleaved, enzymatically or non-enzymatically, reductively, oxidatively, or hydrolytically, to reveal the anionic group. An anionic group can be esterified with moieties (e.g., acyloxymethyl esters) which are cleaved to reveal an intermediate compound which subsequently decomposes to yield the active compound. A discussion of the use of prodrugs is provided by T. Higuchi and W. Stella, "Pro-drugs as Novel Delivery Systems," Vol. 14 of the A.C.S. Symposium Series, and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987.

The term "treating" or "treatment" of a state, disease, disorder or condition includes:

(1) preventing or delaying the appearance of clinical symptoms of the state, disease, disorder or condition developing in a subject that may be afflicted with or predisposed to the state, disease, disorder or condition but does not yet experience or display clinical or subclinical symptoms of the state, disease, disorder or condition;

(2) inhibiting the state, disease, disorder or condition, i.e., arresting or reducing the development of the state, disease, disorder or condition or at least one clinical or subclinical symptom thereof; or

(3) relieving the state, disease, disorder or condition, i.e., causing regression of the state, disease, disorder or condition or at least one of its clinical or subclinical symptoms.

The term "subject" includes mammals (especially humans) and other animals, such as domestic animals (e.g., household pets including cats and dogs) and non-domestic animals (such as wildlife). The term "therapeutically effective amount" means the amount of a compound that, when administered to a subject for treating a state, disease, disorder or condition, is sufficient to effect such treatment. The "therapeutically effective amount" will vary depending on the compound, the state, disease, disorder or condition and its severity and the age, weight, physical condition and responsiveness of the subject receiving treatment.

The compounds of the present invention may form salts. Non-limiting examples of pharmaceutically acceptable salts forming part of this invention include salts derived from inorganic bases, salts of organic bases, salts of chiral bases, salts of natural amino acids and salts of non-natural amino acids. Certain compounds of the present invention are capable of existing in stereo isomeric forms (e.g., diastereomers, enantiomers, racemates, and combinations thereof). With respect to the overall compounds described by the formula (I), the present invention extends to these stereo isomeric forms and to mixtures thereof. To the extent prior art teaches synthesis or separation of particular stereoisomers, the different stereo isomeric forms of the present invention may be separated from one another by the methods known in the art, or a given isomer may be obtained by stereo specific or asymmetric synthesis. Tautomeric forms and mixtures of compounds described herein are also contemplated.

Pharmaceutically acceptable solvates includes hydrates and other solvents of crystallization (such as alcohols). The compounds of the present invention may form solvates with low molecular weight solvents by methods known in the art.

PHARMACEUTICAL COMPOSITIONS

The pharmaceutical compositions provided in the present invention include at least one compound described herein and at least one pharmaceutically acceptable excipient (such as a pharmaceutically acceptable carrier or diluent). Specifically, the contemplated pharmaceutical compositions include a compound(s) described herein in an amount sufficient to treat viral infection in a subject.

The compound of the present invention may be associated with a pharmaceutically acceptable excipient (such as a carrier or a diluent) or be diluted by a carrier, or enclosed within a carrier which can be in the form of a capsule, sachet, or other container.

Examples of suitable carriers include, but are not limited to, water, salt solutions, alcohols, polyethylene glycols, polyhydroxyethoxylated castor oil, peanut oil, olive oil, gelatin, lactose, terra alba, sucrose, dextrin, magnesium carbonate, sugar, cyclodextrin, amylose, magnesium stearate, talc, gelatin, agar, pectin, acacia, stearic acid or lower alkyl ethers of cellulose, silicic acid, fatty acids, fatty acid amines, fatty acid monoglycerides and diglycerides, pentaerythritol fatty acid esters, polyoxyethylene, hydroxymethylcellulose and polyvinylpyrrolidone.

The carrier or diluent may include a sustained release material, such as, for example, glyceryl monostearate or glyceryl distearate, alone or mixed with a wax.

The pharmaceutical composition may also include one or more pharmaceutically acceptable auxiliary agents, wetting agents, emulsifying agents, suspending agents, preserving agents, salts for influencing osmotic pressure, buffers, sweetening agents, flavoring agents, colorants, or any combination of the foregoing. The pharmaceutical composition of the invention may be formulated so as to provide quick-, sustained-, or delayed-release of the active ingredient after administration to the subject by employing procedures known in the art.

The pharmaceutical compositions described herein may be prepared, e.g., as described in Remington: The Science and Practice of Pharmacy, 20 ^th Ed., 2003 (Lippincott Williams & Wilkins). For example, the active compound can be mixed with a carrier, or diluted by a carrier, or enclosed within a carrier, which may be in the form of an ampule, capsule, or sachet. When the carrier serves as a diluent, it may be a solid, semi- solid, or liquid material that acts as a vehicle, excipient, or medium for the active compound.

The pharmaceutical compositions may be, for example, capsules, tablets, aerosols, solutions, suspensions, liquids, gels, or products for topical application.

The route of administration may be any route which effectively transports the active compound to the appropriate or desired site of action. Suitable routes of administration include, but are not limited to, oral, nasal, pulmonary, buccal, subdermal, intradermal, transdermal, parenteral, rectal, depot, subcutaneous, intravenous, intraurethral, intramuscular, intranasal, ophthalmic (such as with an ophthalmic solution) or topical (such as with a topical ointment). The oral route is specifically suitable.

Solid oral formulations include, but are not limited to, tablets, capsules (soft or hard gelatin), dragees (containing the active ingredient in powder or pellet form), troches and lozenges. Tablets, dragees, or capsules having talc and/or a carbohydrate carrier or binder or the like are particularly suitable for oral application. Exemplary carriers for tablets, dragees, or capsules include lactose, cornstarch, and/or potato starch. A syrup or elixir can be used in cases where a sweetened vehicle can be employed.

A typical tablet that may be prepared by conventional tableting techniques.

Liquid formulations include, but are not limited to, syrups, emulsions, soft gelatin and sterile injectable liquids, such as aqueous or non-aqueous liquid suspensions or solutions. For parenteral application, particularly suitable are injectable solutions or suspensions, specifically aqueous solutions with the active compound dissolved in polyhydroxylated castor oil.

METHODS OF SCREENING

Antiviral HIV activity and cytotoxicity of compounds according to present invention can be measured in parallel by following the methods published in the literature.

The cytotoxic effect of compounds can be analyzed by measuring the proliferation of cells using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazlium bromide (MTT) staining. Cells (5 x 10 cells /well) will be incubated in in 96 well plates in the presence or absence of compounds. At the end of treatment, 20μ1 of MTT (5mg/ml in PBS) will be added to each well and incubated for an additional 4 hours at 37°C. The purple -blue MTT formazan precipitate will be dissolved in a triplex reagent containing 10% SDS, 5% isobutanol and 10 mmol/lit HC1. The activity of mitochondria, reflecting cellular growth and viability, will be evaluated by measuring the optical density at 570 nm on micro titer plate.

Action of compounds on replication of HIV in Sup-Tl cells can be determined by the method published by Roda Rani et al., 2006 (Archives of Biochemistry and Biophysics, Volume 456, Issue 1, 1 December 2006, Pages 79-92).

Briefly, lxlO ⁶ Sup-Tl cells with 100% cell viability will be seeded in RPMI 1640, 0.1% FBS four 12 well plates. Increasing concentrations of Epap-1 peptides will be added to the cells and will be infected with HIV1 93 I 101 each at final concentration of virus equivalent to 2 ng of p24 per ml. The infected cells will be incubated at 37 C and 5% C02 incubator for 2 hours. After 2hrs the cells will be pelleted at 350 g for 10 min, supernatant will be discarded and cell will be held with RPMI 1640 containing 10% FBS. The cells will be resuspended in the same medium with increasing concentrations of Epap-1 peptides and will be incubated for 96 hours. The cells will be supplemented with peptides at every 24 hours.The supernatants will be collected after 96 hours and analyzed using P24 antigen capture assay kit (SAIC Fredrick). The infection in the absence of Epap-1 will be considered to be 0% inhibition Azidothymidine (AZT) will be taken as positive control.

Action of compound on virus entry and quantification of virus entered can be done in terms of GFP expression by the following the methods published J. Virol. 72, 6988 (1998) by in Cecilia et al., and Analytical Biochemistry Volume 360, Issue 2, 15 January 2007, Pages 315-317 (Dyavar S. Ravi and Debashis Mitra).

Briefly, cells will be seeded in to wells of 24 well plates 1 day prior to the experiment. The cells will be transfected with Tat-reporter. The virus inoculum will be adjusted to 1,000- 4,000 TCID 50/ ml in assay medium (DMEM,10%FCS,glutamine and antibiotics), 50 μΐ aliquots will be incubated with serial dilutions of compounds (50 μΐ ) for lhr at 37°C. The reporter expression will be quantified at appropriate time calculated inhibitory doses referrers to the concentration of these agents in this preincubation mixture.

Other relevant references useful for screening antiviral HIV activity are: Averett, D.R.1989. Anti-HIV compound assessment by two novel high capacity assays. J. Virol. Methods 23: 263-276; Schwartz, O., et al.1998; A rapid and simple colorimeric test fror the study of anti HIV agents. AIDS Res. and Human Retroviruses, 4(6):441-447; Daluge, S. M., et al. 1994. 5-Chloro-2',3'-deoxy-3'fluorouridine (935U83), a selective anti human immunodeficiency virus agent with an improved metabolic and toxicological profile; Antimicro. Agents and Chemotherapy, 38(7): 1590-1603; H.Mitsuya and S.Border, Inhibition of the in vitro infectivity and cytopathic effect of human T-lympho tropic virus type lymphadenopathy-associated virus (HLTV-III/LAV) by 2,3'-dideoxynucleosides, Proc. Natl. Acad. Sci. USA,83, 1911-15(1986); Pennington et al., Peptides 1990; Meek T.D et al., Inhibition of HIV-1 protease in infected T-limphocytes by synthetic peptide analogues, Nature, 343, p90 (1990); Weislow et al., J. Natl. Cancer Inst. 81, 577-586, 1989; T. Mimoto et al ., J. Med. Chem., 42, 1789-1802, 1999; Uckun et al 1998, Antimicobial Agents and Chemotherapy 42:383; for P24 antigen assay Erice et al., 1993, Antimicrob. Ag. Chemotherapy 37: 385-383; Koyanagi et al., Int. J. Cancer, 36, 445-451, 1985; Balzarini et al. AIDS (1991), 5, 21-28; Connor et al., Journal of virology, 1996, 70, 5306-5311; Popik et al., Journal of virology, 2002, 76, 4709-4722; Harrigton et al., Journal of Virology Methods, 2000, 88, 111-115; Roos et al.,Virology 2000, 273, 307-315; Fedyuk N.V. et al; Problems of Virology 1992, (3)P135; Mosmann T, December 1983, Journal of immunological methods, 65 (1-2), 55-63 ; SPC Cole, cancer chemotherapy and Pharmacology, 1986, 17, 259-263.

METHODS OF TREATMENT

The present invention provides compounds and pharmaceutical compositions thereof that are useful in the treatment of diseases, conditions and/or disorders mediated by viral infections. The connection between therapeutic effect and antiviral is illustrated. For example, PCT publication Nos. WO 01//07646, WO 01/65957, or WO 03/037908; US publication Nos. US 4,598,095 or US 2002/0068757; EP publication Nos. EP 0989862 or EP 0724650; Bioorganic & Medicinal Chemistry Letters, 16, (6), 1712-1715, 2006; and references cited therein, all of which are incorporated herein by reference in their entirety and for the purpose stated.

The present invention further provides a method of treating a disease, condition and/or disorder mediated by viral infections in a subject in need thereof by administering to the subject a therapeutically effective amount of a compound or a pharmaceutical composition of the present invention.

Diseases, conditions, and/or disorders that are mediated by viral infections are believed to include, but are not limited to, HIV infection, HBV, HCV, a retroviral infection genetically related to HIV, AIDS, inflammatory disease, respiratory disorders (including adult respiratory distress syndrome (ARDS), bronchitis, chronic bronchitis, chronic obstructive pulmonary disease, cystic fibrosis, asthma, emphysema, rhinitis and chronic sinusitis), inflammatory bowel disease (including Crohn's disease and ulcerative colitis), multiple sclerosis, rheumatoid arthritis, graft rejection (in particular but not limited to kidney and lung allografts), endometriosis, type I diabetes, renal diseases, chronic pancreatitis, inflammatory lung conditions, chronic heart failure and bacterial infections (in particular but not limited to tuberculosis).

The compounds of the present invention can obtain more advantageous effects than additive effects in the prevention or treatment of the above diseases when using suitably in combination with the available drugs. Also, the administration dose can be decreased in comparison with administration of either drug alone, or adverse effects of co administrated drugs other than antiviral can be avoided or declined.

METHODS OF PREPARATION

The compounds described herein may be prepared by techniques known in the art. In addition, the compounds described herein may be prepared by following the reaction sequence as depicted in Scheme- 1 to 2. Further, in the following schemes, where specific bases, acids, reagents, solvents, coupling agents, etc., are mentioned, it is understood that other bases, acids, reagents, solvents, coupling agents etc., known in the art may also be used and are therefore included within the present invention. Variations in reaction conditions, for example, temperature and/or duration of the reaction, which may be used as known in the art, are also within the scope of the present invention. All the stereoisomers of the compounds in these schemes, unless otherwise specified, are also encompassed within the scope of this invention.

In a further aspect, the compounds of the present invention can also contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the present invention also embraces isotopically-labeled variants of the present invention which are identical to those recited herein, but for the fact that one or more atoms of the compound are replaced by an atom having the atomic mass or mass number different from the predominant atomic mass or mass number usually found in nature for the atom. All isotopes of any particular atom or element as specified are contemplated within the scope of the compounds of the invention, and their uses. Exemplary isotopes that can be incorporated in to compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, sulfur, fluorine, chlorine and iodine, such as H ("D"), ³ H, ⁿ C, ¹³ C, ¹⁴ C, ¹³ N, ¹⁵ N, ¹⁵ 0, ¹⁷ 0, ¹⁸ 0, ³² P, ³³ P, ³⁵ S, ¹⁸ F, ³⁶ C1, ¹²³ I and ¹²⁵ I. Particular isotopes are -CD ₃ or -C(D ₂ )-. Isotopically labeled compounds of the present inventions can generally be prepared by following procedures analogous to those disclosed in the Schemes and/or in the Examples herein below, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

Compounds of the present invention can be synthesized from naturally occurring Betulinic acid or betulinal. Key intermediates required for synthesizing analogues are either commercially available, or can be prepared by the methods published in the literature. For example, the key intermediates in the present invention were prepared by modifying the procedures published in Journal of organic chemistry 2013, 78, 1463-1470; Organic Letters, 2009, 11, 3270-3273; Organic Letters, 2007, 9, 3675-3678; Bio Organic Medicinal Chemistry Letters, 2011, 21, 562-568.

Scheme-1

The compounds of formula 6 (R ₇ and R ₈ are same as defined above and g & gi are substituents such as alkyls, halides or the like) can be prepared as described in scheme 1. The amino acid compounds of formula 1 can be converted to N-protected compounds of formula 2 in the presence of protecting groups such as Di-tert-butyl-dicarbonate [(BOC) ₂ 0], benzyl, Benzyl chloro formate (CbzCl) or the like in the presence of mild bases such as sodium bicarbonate (NaHC0 ₃ ), sodium carbonate (Na ₂ C0 ₃ ), potassium carbonate (K ₂ C0 ₃ ) or the like in the presence of solvents such as 1,4-dioxane, water or the like. The N-protected compounds of formula 2 can be coupled with the bromo compounds of formula 3 to get the O-protected compounds of formula 4 in the presence of suitable coupling agents such as diisopropylethylamine (DIPEA), triethylamine (TEA), potassium carbonate (K ₂ C0 ₃ ) or the like in the solvents such as N,N-Dimethylformamide (DMF), dichloromethane (DCM), dichloroethane, or the like. The O-protected compounds of formula 4 can be cyclized to give the cyclic compounds of formula 5 in the presence of ammonium acetate, or the like in the solvents such as toluene, xylene, or the like. The cyclic compounds (BOC protected) of formula 5 can be deprotected in the presence deprotecting reagents such as HC1, trifluoro acetic acid (TFA), or the like in the solvents such as 1,4-dioxane, dichloromethane (DCM), ethyl acetate (EtOAc) or the like to give the compounds of formula 6.

Scheme - 2

[Formula (I), when Rg is H, Y=0, Z _j is phenyl, Z ₂ & Z ₃ are H]

^~ y ^{~ c} ^Hy ^; is imdazole

The compounds of formula (I) (wherein, Ri, R ₂ , R ₃ , R ₄ , R5, R ₆ , R7, Rg, g, gi and X are same as defined above) can be prepared as described in Scheme 2. The halide compounds of formula 7 (as described in PCT publication WO 2013/160810 A2) can be converted to the C28-amide compounds of formula 8 with their corresponding amine compounds of formula 6 in the presence of suitable bases such as triethylamine (TEA), N,N-diisopropylethylamine (DIPEA), pyridine or the like in the solvents such as Ν,Ν-dimethylformamide (DMF), dichloromethane (DCM), tetrahydrofuran (THF) or the like. The C28-amide compounds (C3 protected) of formula 8 can be deprotected in the presence of bases such as potassium carbonate (K ₂ C0 ₃ ), sodium hydroxide (NaOH), ammonia or the like in the solvents such as methanol: tetrahydrofuran, methanol: water, methanol or the like to give the C3 -hydroxy compounds of formula 9. The C3-hydroxy compounds of formula 9 can be reacted with corresponding acid anhydrides, half protected diacids or their mixed anhydrides or acid chlorides to give the corresponding acid compounds of the present invention formula (I) in the presence of bases such as triethyl amine (TEA), 4-dimethylaminopyridine, diisopropyl ethylamine (DIPEA), pyridine or the like in the solvents such as dichloromethane (DCM), toluene, ethylacetate (EtOAc), tetrahydrofuran (THF) or the like.

The abbreviations used in the entire specification may be summarized herein below with their particular meaning: DIPEA (Ν,Ν-Diisopropylethylamine); °C (degree Celsius); δ (delta); ppm (parts per million); % (percentage); DMSO-d ₆ (Deuterated DMSO); d (Doublet); dd (Doublet of doublet); EtOH (Ethanol); EtOAc (Ethyl acetate); g or gr (gram); H or H ₂ (Hydrogen); HC1 (Hydrochloric acid); h or hr. (Hours); HATU (0-(7-Azabenzotriazol-l-yl)- Ν,Ν,Ν',Ν'-tetramethyluroniumhexafluoro phosphate); Hz (Hertz); HPLC (High-performance liquid chromatography); mmol (Milli mol); M (Molar); ml (Millilitre); mg (Milli gram); m (Multiplet); mm (Millimetre); MHz (Megahertz); ESI-MS (Electron spray Ionization Mass spectra); min (Minutes); mM (Milli molar); NaOH (Sodium hydroxide); N ₂ (Nitrogen); NMR (Nuclear magnetic resonance spectroscopy); S (Singlet); TEA (Triethyl amine); TLC (Thin Layer Chromatography); THF (Tetrahydrofuran); tert (Tertiary), t (Triplet); IC (Inhibitory concentration), nM (Nano molar); pH (Pouvoir hydrogen); (Boc) ₂ 0 (Di-tert-butyl dicarbonate); DCM (dichloromethane); DMF (Ν,Ν-dimethyl formamide); DMAP (4- (Dimethylamino)pyridine); eq (equivalent); Ltr or L (Liter); CDC1 ₃ (Deuterated chloroform); J (Coupling constant); AcOH (Acetic acid); ABq (AB quartet); K ₂ CQ (potassium carbonate); Cs ₂ C0 ₃ (Cesium carbonate); NaHC0 ₃ (Sodium bicarbonate); Na ₂ S0 ₄ (Sodium sulphate); MeOH (methanol); EDCI (l-Ethyl-3-(3-dimethylaminopropyl)carbodiimide); HOBt (1-Hydroxybenzotriazole); brs (broad singlet); DCC (Ν,Ν'-dicyciohexyicarbodiimide): Pd/C (palladium in carbon) and Pd(OAc) ₂ (palladium acetate).

EXPERIMENTAL

The present invention is further illustrated by the following examples, which are not to be construed in any way as imposing limitations upon the scope of this disclosure, but rather are intended to be illustrative only. On the contrary, it is to be clearly understood that resort may be had to various other embodiments, modifications, and equivalents thereof which, after reading the description herein, may suggest themselves to one of ordinary skill in the art without departing from the spirit of the present invention. Thus, the skilled artisan will appreciate how the experiments and examples may be further implemented as disclosed by variously altering the following examples, substituents, reagents, or conditions.

INTERMEDIATES

Intermediate- 1 : Preparation of (S)-l-(5-phenyl-lH-imidazol-2-yl)ethan-l-aminium chloride

Synthesis of ( tert-butoxycarbonyl)-L-alanine

BocHN

OH

To a stirred solution of L-alanine (25.0 gr, 280.61 mmol) in 1,4-dioxane (125 mL) added 2N NaOH solution (125 mL), (Boc) ₂ 0 (91.7 gr, 420.0 mmol) at 0°C and stirred the reaction mixture for about 12 hours at room temperature. After complete conversion of starting material, the reaction mixture was washed with EtOAc (200 mL) to remove the impurities, then aqueous part was acidified with IN HC1 (pH-2-3) and extracted with CH ₂ C1 ₂ (2x300 mL). The combined organic extracts were washed with water, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 30% EtOAc: n-Hexane as an eluent to afford the desired product (50.0 g, yield: 94.3%) as a light yellow liquid. 1H MR (300 MHz, DMSO): δ 12.39 (bs, 1H), 7.11 (d, J = 7.5 Hz, 1H), 3.94-3.86 (m, 1H), 1.38 (s, 9H), 1.21 (d, J = 7.2 Hz); ES Mass: [M+23] ⁺ 212.01.

Step 2: Synthesis of 2-oxo-2-phenylethyl (tert-butoxycarbonyl)-L-alaninate

To a stirred solution of (tert-butoxycarbonyl)-L- alanine (step 1, 52.0 gr, 273.7 mmol) in CH ₂ C1 ₂ (520 mL), added DIPEA (100.9 mL, 547.26 mmol), phenacylbromide (17.64 gr, 88.64 mmol) at 0°C and stirred the reaction mixture for about 12 hours at room temperature. After complete conversion of starting material, the reaction mixture was extracted with CH ₂ C1 ₂ (2x200 mL). The combined organic extracts were washed with water, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 20% EtOAc: n-Hexane as an eluent to afford the desired product (60.0 g, yield: 71.4%) as a yellow color solid. 1H MR (300 MHz, DMSO): δ 8.02-7.96 (m, 2H), 7.72-7.67 (m, 1H), 7.59-7.54 (m, 2H), 7.38 (d, J = 7.2 Hz, 1H), 5.5 (ABq, J = 17.5 Hz, 2H), 4.95 (s, 1H), 4.19-4.14 (m, 1H), 1.38-1.35 (m, 12H); ES Mass: [M+23] ⁺ 330.02.

Step 3: Synthesis of tert-butyl (S)-(l-(5-phenyl-lH-imidazol-2-yl)ethyl)carbamate

To a stirred solution of 2-oxo-2-phenylethyl (tert-butoxycarbonyl)-L-alaninate (step 2, 40.0 gr, 130.29 mmol) in toluene (400 mL), added NH ₄ OAc (80.3 gr, 1042.0 mmol) and stirred the reaction mixture for about 12 hours at 90°C. After complete conversion of starting material, the reaction mixture was extracted with EtOAc (2x400 mL). The combined organic extracts were washed with water, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 50% EtOAc: n-Hexane as an eluent to afford the desired product (30.0 g, yield: 80.2%) as a brown color solid. 1H MR (300 MHz, DMSO): δ 11.78 (bs, 1H), 7.78-7.3 (m, 2H), 7.49 (s, 1H), 7.41-7.29 (m, 2H), 7.23-7.08 (m, 2H), 4.72-4.67 (m, 1H), 1.41-1.39 (m, 12H); ES Mass: [M+l] ⁺ 288.1.

Step 4: Synthesis of (S)-l-(5-phenyl-lH-imidazol-2-yl)ethan-l-aminium chloride

To a stirred solution of tert-butyl (S)-(l-(5-phenyl-lH-imidazol-2-yl)ethyl)carbamate (step 3, 15.0 g, 52.23 mmol, 1.0 eq) in CH ₂ C1 ₂ (30 ml) was added 4M HCl in dioxane (150 mL).The reaction mixture was stirred at room temperature for about 3 hours. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was concentrated under reduced pressure to get the residue. (11.7 gr, yield: 100%) and used directly for next step without further purification.

Intermediate-2: Preparation of (S)-2-methyl-l-(5-phenyl-lH-imidazol-2-yl)propan-l- aminium chloride

Step 1: Synthesis of (tert-butoxycarbonyl)-L-valine

o

BocHN. A. OH

To a stirred solution of L-valine (5.0 gr, 42.7 mmol) in 1,4-dioxane (25 mL) added 2N NaOH solution (25 mL), (Boc) ₂ 0 (13.97 gr, 64.1 mmol) at 0°C and stirred the reaction mixture for about 12 hours at room temperature. After complete conversion of starting material, the reaction mixture was washed with EtOAc (200 mL) to remove the impurities, then aqueous part was acidified with IN HCl (P ^H -2-3) and extracted with CH ₂ C1 ₂ (2x200 mL). The combined organic extracts were washed with water, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 30% EtOAc: n-Hexane as an eluent to afford the desired product (9.0 g, yield: 96.8%) as an off white solid. 1H NMR (300 MHz, CDC1 ₃ ): δ 5.02 (d, J = 8.7 Hz, 1H), 4.28-4.23 (m, 1H), 4.09-4.05 (m, 1H), 2.24-2.18 (m, 1H), 1.45 (s, 9H), 1.0 (d, J = 6.9 Hz, 3H), 0.93 (d, J = 6.9 Hz, 3H); ES Mass: [M+23] ⁺ 240.14.

Step 2: Synthesis of 2-oxo-2-phenylethyl (tert-butoxycarbonyl)-L-valinate

To a stirred solution of (tert-butoxycarbonyl)-L-valine (step 1, 10.0 gr, 46.08 mmol) in CH ₂ C1 ₂ (100 mL) added DIPEA (26.1 mL, 92.0 mmol) and phenacylbromide (11.0 gr, 55.2 mmol) at 0°C and stirred the reaction mixture for about 12 hours at room temperature. After complete conversion of starting material, the reaction mixture was extracted with CH ₂ C1 ₂ (2x200 mL). The combined organic extracts were washed with water, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 20% EtOAc: n-Hexane as an eluent to afford the desired product (15.0 g, yield: 97.0%) as a yellow color solid. 1H NMR (300 MHz, CDC1 ₃ ): δ 7.97- 7.89 (m, 2H), 7.65-7.59 (m, 1H), 7.52-7.47 (m, 2H), 5.53 (d, J = 16.5 Hz, 1H), 5.28 (d, J = 16.5 Hz, 1H), 5.05 (d, J = 9.0 Hz, 1H), 4.43-4.38 (m, 1H), 2.38-2.29 (m, 1H), 1.46 (s, 9H), 1.07 (d, J = 6.9 Hz, 3H), 1.02 (d, J = 6.9 Hz, 3H); ES Mass: [M+l] ⁺ 336.2.

Step 3: Synthesis of tert-butyl (S)-(2-methyl-l-(5-phenyl-lH-imidazol-2-yl)propyl)carbamate

To a stirred solution of 2-oxo-2-phenylethyl (tert-butoxycarbonyl)-L-valinate (step 2, 15.0 gr, 44.7 mmol) in toluene (150 mL) added NH ₄ OAc (34.8 gr, 447.0 mmol) and stirred the reaction mixture for about 12 hours at 90°C. After complete conversion of starting material, the reaction mixture was extracted with EtOAc (2x200 mL). The combined organic extracts were washed with water, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 30% EtOAc: n-Hexane as an eluent to afford the desired product (11.0 g, yield: 78.0%) as a brown color solid. 1H NMR (300 MHz, CDC1 ₃ ): δ 11.76 (s, 1H), 7.75-7.72 (m, 2H), 7.5 (s, 1H), 7.34-7.27 (m, 2H), 6.96-6.94 (m, 1H), 4.43-4.38 (m, 1H), 2.12-2.06 (m, 1H), 1.39 (s, 9H),

0.89 (d, J = 6.6 Hz, 3H), 0.76 (d, J = 6.6 Hz, 3H); ES Mass: [M+l] ⁺ 316.24.

Step 4: Synthesis of (S)-2-methyl-l -(5-phenyl-lH-imidazol-2-yl)propan-l -aminium chloride

To a stirred solution of tert-butyl (S)-(2-methyl-l-(5-phenyl-lH-imidazol-2- yl)propyl)carbamate (step 3, 4.0 g, 12.69 mmol, 1.0 eq) in CH ₂ C1 ₂ (8 ml) was added 4M HCl in dioxane (32 mL). The reaction mixture was stirred at room temperature for about 3 hours. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was concentrated under reduced pressure to get the residue (2.5 g, yield: 78.6%) and used directly for next step without further purification.

Intermediate- 3: Preparation of 2-(5-phenyl-lH-imidazol-2-yl)propan-2-aminium chloride

Step 1: Synthesis of2-((tert-butoxycarbonyl)amino)-2-methylpropanoic acid

To a stirred solution of 2-carboxypropan-2-aminium chloride (15.0 gr, 145.63 mmol) in 1,4-dioxane (75 mL), added 2N NaOH solution (75 mL), (Boc) ₂ 0 (47.62 gr, 218.44 mmol) at 0°C and stirred the reaction mixture for about 12 hours at room temperature. After complete conversion of starting material, the reaction mixture was washed with EtOAc (200 mL) to remove the impurities, then aqueous part was acidified with IN HCl (P -2-3) and extracted with CH ₂ C1 ₂ (2x200 mL). The combined organic extracts were washed with water, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 30% EtOAc: n-Hexane as an eluent to afford the desired product (15.0 g, yield: 50.74%) as an off white solid. 1H MR (300 MHz, DMSO): δ 12.18 (s, 1H), 7.05 (s, 1H), 1.36 (s, 9H), 1.29 (s, 6H); ES Mass: [M+23] ⁺ 226.06. Step 2: Synthesis of 2-oxo-2-phenylethyl 2-((tert-butoxycarbonyl)amino)-2-methyl propanoate

To a stirred solution of 2-((tert-butoxycarbonyl)amino)-2-methylpropanoic acid (step 1, 15.0 gr, 73.89 mmol) in CH ₂ C1 ₂ (150 mL) added DJPEA (40.85 mL, 316.6 mmol), phenacylbromide (17.64 gr, 88.64 mmol) at 0°C and stirred the reaction mixture for about 12 hours at room temperature. After complete conversion of starting material, the reaction mixture was extracted with CH ₂ C1 ₂ (2x200 mL). The combined organic extracts were washed with water, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 20% EtOAc: n-Hexane as an eluent to afford the desired product (22.3 g, yield: 95.05%) as a yellow color solid. 1H MR (300 MHz, CDC1 ₃ ): δ 7.92-7.89 (m, 2H), 7.64-7.59 (m, 1H), 7.52-7.47 (m, 2H), 5.39 (s, 2H), 1.63 (s, 6H), 1.44 (s, 9H); ES Mass: [M+23] ⁺ 343.87.

Step 3: Synthesis of tert-butyl (2-(5-phenyl-lH-imidazol-2-yl)propan-2-yl)carbamate

To a stirred solution of 2-oxo-2-phenylethyl 2-((tert-butoxycarbonyl)amino)-2- methylpropanoate (step 2, 22.3 gr, 69.68 mmol) in toluene (200 mL) added NH ₄ OAc (37.6 gr, 487.81 mmol) and stirred the reaction mixture for about 12 hours at 90°C. After complete conversion of starting material, the reaction mixture was extracted with EtOAc (2x200 mL). The combined organic extracts were washed with water, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 30% EtOAc: n-Hexane as an eluent to afford the desired product (17.0 g, yield: 81.3%) as a brown color solid. 1H NMR (300 MHz, CDC1 ₃ ): δ 10.63 (bs, 1H), 7.63 (bs, 2H), 7.39-7.34 (m, 2H), 7.26-7.22 (m, 2H), 5.14 (s, 1H), 1.77 (s, 6H), 1.44 (s, 9H); ES Mass: [M+l] ⁺ 302.27.

Step 4: Synthesis of2-(5-phenyl-lH-imidazol-2-yl)propan-2-aminium chloride

To a stirred solution of tert-butyl (2-(5-phenyl-lH-imidazol-2-yl)propan-2- yl)carbamate (step 3, 17.0 g, 56.47 mmol, 1.0 eq) in CH ₂ C1 ₂ (34 ml) was added 4M HCl in dioxane (170 mL). The reaction mixture was stirred at room temperature for about 3 hours. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was concentrated under reduced pressure to get the residue (13.4 gr, yield: 100%) and used directly for next step without further purification.

Synthesis of l-(( tert-butoxycarbonyl)amino )cyclobutane-l -carboxylic acid

To a stirred solution of 1-aminocyclobutane-l -carboxylic acid (2 g, 17 mmol) in 1,4- dioxane: H ₂ 0 (20: 20 mL), were added NaHC0 ₃ (4.4 g, 57 mmol) and (Boc) ₂ 0 (4.5 g, 20.4 mmol) at 0°C and stirred the reaction mixture for about 12 hours at room temperature. After complete conversion of starting material, the reaction mixture was washed with EtOAc (30 mL) to remove the impurities, then aqueous layer was acidified with IN HC1 (P -2-3) and extracted with CH ₂ C1 ₂ (2x40 mL). The combined organic extracts were washed with water, brine, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silica gel column chromatography by using 35 % EtOAc: n-Hexane as an eluent to afford the desired product (2.8 g, yield: 75%) as an off white solid. 1H MR (300 MHz, CD ₃ OD): δ 2.62-2.53 (m, 2H), 2.25-2.15 (m, 2H), 2.05-1.97 (m, 2H), 1.42 (s, 9H); ES Mass: 238.04 [M+Na] ⁺ .

Step 2: Synthesis of 2-oxo-2-phenylethyl l-((tert-butoxycarbonyl)amino)cyclobutane-l- carboxylate

To a stirred solution of l-((tert-butoxycarbonyl)amino)cyclobutane-l -carboxylic acid (step 1, 2.8 g, 12 mmol) in CH ₂ C1 ₂ (40 mL), were added DIPEA (4.2 mL, 24 mmol) and 2- bromo-l-phenylethan-l-one (2.6 ml, 13 mmol) at 0°C and stirred the reaction mixture for about 12 hours at room temperature. After complete conversion of starting material, the reaction mixture was extracted with CH ₂ C1 ₂ (2x50 mL). The combined organic extracts were washed with water, brine, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silica gel column chromatography by using 20% EtOAc: n-Hexane as an eluent to afford the desired product (4 g, yield: 93%) as a yellow colour solid.

1H MR (300 MHz, CDCI ₃ ): δ 7.93-7.90 (m, 2H), 7.64-7.59 (m, 3H), 5.42 (s, 2H), 2.86-2.77

(m, 2H), 2.41-2.30 (m, 2H), 2.10-2.04 (m, 2H), 1.44 (s, 9H). ES Mass: 356.11 [M+Na] ⁺ . Step 3: Synthesis of tert-butyl (l-(5-phenyl-lH-imidazol-2-yl)cyclobutyl)carbamate

To a stirred solution of 2-oxo-2-phenylethyl l-((tert-butoxycarbonyl)amino) cyclobutane-l-carboxylate (step 2, 2 g, 6 mmol) in toluene (20 mL), was added NH ₄ OAc (4.8 g, 62 mmol) and stirred the reaction mixture for about 12 hours at 90°C. After complete conversion of starting material, the reaction mixture was extracted with EtOAc (2x50 mL). The combined organic extracts were washed with water, brine, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silica gel column chromatography by using 30% EtOAc: n-Hexane as an eluent to afford the desired product (1 g, yield: 54%) as a brown colour solid. 1H MR (300 MHz, CDC1 ₃ ): δ 7.66 (d, J = 7.5 Hz, 2H), 7.39 (dd, J = 7.5 Hz, 7.5 Hz, 2H), 7.29-7.23 (m, 2H), 5.48 (s, 1H), 2.97-2.88 (m, 2H), 2.44-2.34 (m, 2H), 2.13-1.98 (m, 2H), 1.45 (s, 9H); ES Mass: 314.18 [M+H] ⁺ .

Step 4: Synthesis of 1 -(5 -phenyl- lH-imidazol-2-yl)cyclobutan-l -amine hydro chloride

To a stirred solution of tert-butyl (l-(5-phenyl-lH-imidazol-2-yl)cyclobutyl) carbamate (step 3, 0.75 g, 2.4 mmol) in dioxane (2 ml), was added 6N HC1 in dioxane (8 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (0.59 g, yield: 100%). Next reaction was carried out without any further purification.

Intermediate- 5: l-(5-(4-fluorophenyl)-lH-imidazol-2-yl)cyclobutan-l-amine hydrochloride

Step 1: Synthesis of 2-(4-fluorophenyl)-2-oxoethyl l-((tert-butoxycarbonyl)amino) cyclobutane-1 -carboxylate

To a stirred solution of l-((tert-butoxycarbonyl)amino)cyclobutane-l-carboxylic acid (2 g, 9 mmol) in CH ₂ C1 ₂ (30 mL), were added DIPEA (3.6 mL, 27 mmol) and 2-bromo-l-(4- fluorophenyl)ethan-l-one (2 ml, 13 mmol) at 0°C and stirred the reaction mixture for about 12 hours at room temperature. After complete conversion of starting material, the reaction mixture was extracted with CH ₂ CI ₂ (2x50 mL). The combined organic extracts were washed with water, brine, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silica gel column chromatography by using 20% EtOAc: n- Hexane as an eluent to afford the desired product (2.6 g, yield: 82%) as a yellow colour solid. 1H MR (300 MHz, CDCI ₃ ): δ 7.97-7.93 (m, 2H), 7.17 (dd, J = 8.4 Hz, 8.4 Hz, 2H), 5.38 (s, 2H), 2.85-2.76 (m, 2H), 2.41-2.30 (m, 2H), 2.12-2.07 (m, 2H), 1.44 (s, 9H); ES Mass: 374.09 [M+Na] ⁺ .

Step 2: Synthesis of tert-butyl (l-(5-(4-fluorophenyl)-lH-imidazol-2-yl)cyclobutyl)carbamate

To a stirred solution of 2-(4-fluorophenyl)-2-oxoethyl l-((tert-butoxycarbonyl)amino) cyclobutane-l-carboxylate (step 1, 4 g, 11.3 mmol) in toluene (40 mL), was added NH ₄ OAc (9.6 g, 124 mmol) and stirred the reaction mixture for about 12 hours at 90°C. After complete conversion of starting material, the reaction mixture was extracted with EtOAc (2x100 mL). The combined organic extracts were washed with water, brine, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silica gel column chromatography by using 30% EtOAc: n-Hexane as an eluent to afford the desired product (2.1 g, yield: 56%) as a brown colour solid. 1H MR (300 MHz, CDCI ₃ ): δ 7.66 (dd, J = 5.4 Hz, 8.1 Hz, 2H), 7.18 (s, 1H), 7.06 (dd, J = 8.1 Hz, 8.1 Hz, 2H), 5.47 (s, 1H), 2.95-2.86 (m, 2H), 2.40-2.33 (m, 2H), 2.07-2.00 (m, 2H), 1.45 (s, 9H); ES Mass: 332.22 [M+H] ⁺ .

Step 3: Synthesis of l-(5-(4-fluorophenyl)-lH-imidazol-2-yl)cyclobutan-l-amine hydro chloride

To a stirred solution of tert-butyl (l-(5-(4-fluorophenyl)-lH-imidazol-2-yl)cyclobutyl) carbamate (step 2, 0.8 g, 2.4 mmol) in dioxane (4 ml), was added 6N HC1 in dioxane (10 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (0.64 g, yield: 100%). Next reaction was carried out without any further purification.

Intermediate- 6: Preparation of l-(5-phenyl-lH-imidazol-2-yl)cyclopentan-l-amine hydro chloride

Step 1: Synthesis of 1 -((tert-butoxycarbonyl)amino)cyclopentane-l -carboxylic acid

To a stirred solution of 1-aminocyclopentane-l -carboxylic acid (6 g, 46 mmol) in 1,4- dioxane: H ₂ 0 (50: 50 mL), were added NaHC0 ₃ (13 g, 138 mmol) and (Boc) ₂ 0 (13.5 g, 55.8 mmol) at 0°C and stirred the reaction mixture for about 12 hours at room temperature. After complete conversion of starting material, reaction mixture was washed with EtOAc (50 mL) to remove the impurities, then aqueous layer was acidified with IN HC1 (pH=2-3) and extracted with CH ₂ C1 ₂ (2x100 mL). The combined organic extracts were washed with water, brine, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silica gel column chromatography by using 35% EtOAc: n-Hexane as an eluent to afford the desired product (6.7 g, yield: 63%) as an off white solid. 1H MR (300 MHz, CDC1 ₃ ): δ 4.96 (s, 1H), 2.31-2.22 (m, 2H), 1.96-1.90 (m, 2H), 1.85-1.78 (m, 4H), 1.44 (s, 9H); ES Mass: 252.09 [M+Na] ⁺ .

Step 2: Synthesis of 2-oxo-2-phenylethyl l-((tert-butoxycarbonyl)amino)cyclopentane-l- carboxylate

To a stirred solution of l-((tert-butoxycarbonyl)amino)cyclopentane-l -carboxylic acid (step 1, 3 g, 13 mmol) in CH ₂ C1 ₂ (30 mL), were added DIPEA (4.5 mL, 29 mmol) and 2- bromo-l-phenylethan-l-one (2.6 ml, 13.5 mmol) at 0°C and stirred the reaction mixture for about 12 hours at room temperature. After complete conversion of starting material, the reaction mixture was extracted with CH ₂ C1 ₂ (2x50 mL). The combined organic extracts were washed with water, brine, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silica gel column chromatography by using 20% EtOAc: n-Hexane as an eluent to afford the desired product (2 g, yield: 44%) as a yellow colour solid. Step 3: Synthesis of tert-butyl (l-(5-phenyl-lH-imidazol-2-yl)cyclopentyl)carbamate

To a stirred solution of 2-oxo-2-phenylethyl l-((tert-butoxycarbonyl)amino) cyclopentane-l-carboxylate (step 2, 2 g, 5 mmol) in toluene (20 mL), was added NH ₄ OAc (4.8 g, 57 mmol) and stirred the reaction mixture for about 12 hours at 90°C. After complete conversion of starting material, the reaction mixture was extracted with EtOAc (2x50 mL). The combined organic extracts were washed with water, brine, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silica gel column chromatography by using 30% EtOAc: n-Hexane as an eluent to afford the desired product (1 g, yield: 53%) as a brown colour solid. 1H MR (300 MHz, CDC1 ₃ ): δ 7.66 (d, J = 7.2 Hz, 2H), 7.40 (dd, J = 7.2 Hz, 7.2 Hz, 2H), 7.28-7.26 (m, 1H), 7.24 (s, 1H), 5.12 (s, 1H), 2.53- 2.46 (m, 2H), 2.30-2.25 (m, 2H), 1.87-1.85 (m, 4H), 1.46 (s, 9H); ES Mass: 350.12 [M+Na] ⁺ . Step 4: Synthesis of 1 -(5 -phenyl- lH-imidazol-2-yl)cyclopentan-l -amine hydro chloride:

To a stirred solution of tert-butyl (l-(5-phenyl-lH-imidazol-2-yl)cyclopentyl) carbamate (step 3, 0.2 g, 0.61 mmol) in dioxane (2 ml), was added 6N HC1 in dioxane (4 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (0.16 g, yield: 100%). Next reaction was carried out without any further purification.

Intermediate-7: Preparaation of l-(5-phenyl-lH-imidazol-2-yl)cvclopropan-l-amine hydrochloride

Synthesis of l-(( tert-butoxycarbonyl)amino )cyclopropane-l -carboxylic acid

To a stirred solution of 1-aminocyclopropane-l -carboxylic acid (4 g, 39.6 mmol) in 1,4-dioxane: H ₂ 0 (30: 30 mL), were added NaHC0 ₃ (8.3 g, 99 mmol) and (Boc) ₂ 0 (10.4 g,

47.5 mmol) at 0 C and stirred the reaction mixture for 12 h at room temperature. After complete conversion of starting material, reaction mixture was washed with EtOAc (50 mL) to remove the impurities, then aqueous layer was acidified with IN HC1 (pH-2-3) and extracted with CH ₂ CI ₂ (2 X 50 mL). The combined organic extracts were washed with water, brine, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silica gel column chromatography by using 35 % EtOAc: n-Hexane as an eluent to afford the desired product (7.3 g, yield: 91.7%) as an off white solid. 1H NMR (300 MHz, DMSO-d ₆ ): δ 12.26 (brs, 1H), 7.39 (s, 1H), 1.36 (s, 9H), 1.27-1.23 (m, 2H), 0.95-0.92 (m, 2H).

Step 2: Synthesis of 2-oxo-2-phenylethyl 1 -((tert-butoxycarbonyl)amino)cyclopropane-l - carboxylate

BocHN

To a stirred solution of l-((tert-butoxycarbonyl)amino)cyclopropane-l-carboxylic acid (7.3 g, 36.1 mmol) in CH ₂ C1 ₂ (70 mL), were added DIPEA (12.8 mL, 72.2 mmol) and

2-bromo-l-phenylethan-l-one (7.3 ml, 36.5 mmol) at 0 C and stirred the reaction mixture for 12 h at room temperature. After complete conversion of starting material, reaction mixture was extracted with CH ₂ C1 ₂ (2 X 100 mL). The combined organic extracts were washed with water, brine, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure to afford the desired product (11.5 g, Yield: 100 %). Next reaction was carried out without any further purification.

Step 3: Synthesis of tert-butyl (l-(5-phenyl-lH-imidazol-2-yl)cyclopropyl)carbamate

To a stirred solution of 2-oxo-2-phenylethyl l-((tert- butoxycarbonyl)amino)cyclopropane-l -carboxylate (11.5 g, 36.5 mmol) in toluene (120 mL), was added NH ₄ OAc (28.1 g, 365.6 mmol) and stirred the reaction mixture for 12 h at 90

C. After complete conversion of starting material, reaction mixture was extracted with EtOAc (2 X 150 mL). The combined organic extracts were washed with water, brine, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silica gel column chromatography by using 30% EtOAc: n-Hexane as an eluent gave the desired product (8 g, yield: 74.7%) as a brown colour solid. 1H NMR (300 MHz, DMSO-d ₆ ): δ 11.75 (brs, 1H), 7.70-7.68 (m, 2H), 7.59 (s, 1H), 7.37 (s, 1H), 7.33-7.28 (m, 2H), 7.16-7.12 (m, 1H), 1.40 (s, 9H), 1.31-1.30 (m, 2H), 1.09-1.08 (m, 2H). ESI Mass: 300.21 [M+H] ⁺ . Step 4: Synthesis of 1 -(5 -phenyl- lH- -l -amine hydrochloride

To a stirred solution of tert-butyl (l-(5-phenyl-lH-imidazol-2- yl)cyclopropyl)carbamate (0.68 g, 2.2 mmol) in dioxane (2 ml), was added 6N HCl in dioxane (6 ml) and stirred for 2 h. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (0.534 g, Yield: 100 %). Next reaction was carried out without any further purification.

Intermediate- 8: Preparation of (5-phenyl-lH-imidazol-2-yl)methanamine

Step 1: Synthesis of2-((2-oxo-2-phenylethylidene)amino)acetic acid

O

BzCHN .

OH

To a stirred solution of Glycine hydrochloride (10.0 gr, 90.08 mmol) in 1,4- dioxane:H ₂ 0 (1:2, 150 mL) added Na ₂ C0 ₃ (28.6 gr, 270.24 mmol) and CBzCl (15.4 mL,

108.09 mmol) at 0 °C and stirred the reaction mixture for 12 hr at room temperature. After complete conversion of starting material, reaction mixture was washed with EtOAc (200 mL) to remove the impurities, then aq. part was acidified with IN HCl (pH-2-3) and extracted with CH ₂ C1 ₂ (2x200 mL). The combined organic extracts were washed with water, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 70% EtOAc: n-Hexane as an eluent gave the desired product (11.0 g, 58.1 % yield) as light yellow liquid. 1H MR (300 MHz,

CDCI ₃ +DMSO): δ 7.35-7.31 (m, 5H), 6.04 (bs, 1H), 5.12 (s, 2H), 3.91-3.9 (s, 2H).

Step 2: 2-oxo-2-phenylethyl 2-((2-oxo-2-phenylethylidene)amino)acetate

To a stirred solution of 2-((2-oxo-2-phenylethylidene)amino)acetic acid (4.5 gr, 23.6 mmol) in CH ₂ CI ₂ (50 mL) added DIPEA (9.1 gr, 70.7 mmol) and phenacylbromide (7.1 gr, 35.32 mmol) at 0 °C and stirred the reaction mixture for 12 hr at room temperature. After complete conversion of starting material, reaction mixture was extracted with CH ₂ CI ₂ (2x200 mL). The combined organic extracts were washed with water, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 50% EtOAc: n-Hexane as an eluent gave the desired product (5.5 g, 75.3 % yield) as yellow color solid. 1H NMR (300 MHz, DMSO): δ 7.97 (d, J = 7.2 Hz, 2H), 7.82-7.78 (m, 1H), 7.73-7.68 (m, 1H), 7.59-7.54 (m, 2H), 7.39-7.33 (m, 5H), 5.56 (s, 2H), 5.06 (s, 2H), 3.95 (d, J = 6.0Hz, 2H).

Step 3: l-phenyl-2-(((5-phenyl-l midazol-2-yl)methyl)imino)ethan-l-one

BzC

To a stirred solution of 2-oxo-2-phenylethyl 2-((2-oxo-2- phenylethylidene)amino)acetate (5.7 gr, 18.5 mmol) in toluene (50 mL) added NH ₄ OAc (9.9 gr, 129.08 mmol) and stirred the reaction mixture for 12 hr at 90 °C. After complete conversion of starting material, reaction mixture was extracted with EtOAc (2x200 mL). The combined organic extracts were washed with water, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 70% EtOAc: n-Hexane as an eluent gave the desired product (4.5 g, 84.4 % yield) as brown color solid. 1H NMR (300 MHz, DMSO): δ 11.99 (bs, 1H), 7.77-7.71 (m, 3H), 7.49 (s, 1H), 7.39-7.28 (m, 7H), 7.21-7.13 (m, 1H), 5.07 (s, 2H), 4.27 (d, J = 5.7 Hz, 2H), 1.91 (s, 3H).

Step 4: (5-phenyl-lH-imidazol-2-yl)methanamine

To a stirred solution of l-phenyl-2-(((5-phenyl-lH-imidazol-2- yl)methyl)imino)ethan-l-one (4.0 g, 13.03 mmol, 1.0 eq) in CH ₂ C1 ₂ : MeOH (1: 1, 80 ml) was added 10% Pd/C (500 mg). Hydrogen gas was bubbled through the reaction mixture via a balloon for 2 hrs. The resultant mixture was filtered through celite and washed with methanol and the filtrate was concentrated and the crude residue (2.3 g, 96%) was used directly for next step. 1H NMR (300 MHz, DMSO): δ 7.74 (d, J = 7.5 Hz, 2H), 7.53 (s, 1H), 7.37-7.32 (m, 2H), 7.21-7.16 (m, 1H), 3.93 (s, 2H). ES Mass: [M+l] ⁺ 174.06. EXAMPLES

Example- 1: Preparation of (lR,3S)-2,2-dimethyl-3-

((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR, 13bR)-5a,5b,8,8,l la-pentamethyl-3a-(((S)-l- (5-phenyl-lH-imidazol-2-yl)ethyl)carbamoyl)-l-(prop-l-en-2-y l)icosahydro-lH- cyclopentaralchrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxy lic acid

Step 1: Synthesis of (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla- pentamethyl-3a-(((S)-l -(5 -phenyl- lH-imidazol-2-yl)ethyl)carbamoyl)-l -(prop-1 -en-2- yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate

To a stirred solution of (S)-l-(5-phenyl-lH-imidazol-2-yl)ethan-l-aminium chloride (Intermediate- 1, 5.64 g, 25.24 mmol, 1.5 eq) and triethylamine (12.1 mL, 84.14 mmol, 5.0 eq) in CH ₂ C1 ₂ (50 ml) at 0°C was added a solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR, 13aR,13bR)-3a-(chlorocarbonyl)-5a,5b, 8,8,1 la-pentamethyl-1 -(prop-1 -en-2-yl)icosahydro- lH-cyclopenta[a]chrysen-9-yl acetate (prepared as described in WO 2013/160810 A2, 8.7 g, 16.82 mmol, 1.0 eq) in CH ₂ C1 ₂ (50 ml). The reaction mixture was allowed to stir at room temperature for overnight. The reaction mixture was diluted with water and extracted with CH ₂ C1 ₂ (3x200 ml). The combined organic extracts were dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The residue was purified by silicagel column chromatography by using 30% EtOAc: n-Hexane as an eluent to afford the desired product (10.0 g, yield: 89.0%) as a brown color solid. 1H MR (300 MHz, CDC1 ₃ ): δ 11.98 (bs, 1H), 7.72-7.7 (m, 2H), 7.43-7.37 (m, 3H), 7.32-7.23 (m, 2H), 5.42-5.31 (m, 1H), 4.71 (s, 1H), 4.58 (s, 1H), 4.49-4.42 (m, 1H), 2.43-2.31 (m, 2H), 2.28-2.18 (m, 2H), 2.03 (s, 3H), 1.93-1.12 (m, 22H), 1.03-0.74 (m, 20H); ES Mass: [M+l] ⁺ 668.49.

Step 2: Synthesis of (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-9-hydroxy- 5a,5b,8,8,lla-pentamethyl-N-((S)-l-(5-phenyl-lH-imidazol-2-y l)ethyl)-l -(prop-1 -en-2- yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide:

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 5a,5b,8,8,l la-pentamethyl-3a-(((S)-l-(5-phenyl-lH-imidazol-2-yl)ethyl)c arbamoyl)-l-(prop- l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (step 1, 10.0 g, 14.98 mmol, 1.0 eq) in THF (100 ml), Methanol (100 ml) and water (100 mL) was added Lithium hydroxide mono hydrate (5.03 g, 119.85 mmol, 8.0 eq). The reaction mixture was stirred at room temperature for about 12 hours. TLC indicated starting material was consumed and the desired product was observed. The solvents were removed under reduced pressure, the reaction mixture was diluted with water (200 mL) and extracted with ethyl acetate (2X250 mL). The combined organic layers were washed with brine, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude was purified by silicagel column chromatography by using 30% EtOAc: n-Hexane as an eluent to afford the desired product (5.0 g, yield: 53.2%) as a brown color solid. 1H NMR (300 MHz, CDC1 ₃ ): δ 7.66-7.64 (m, 2H), 7.41-7.32 (m, 2H), 7.29-7.18 (m, 2H), 6.68 (bs, 1H), 5.17-5.09 (m, 1H), 4.74 (s, 1H), 4.61 (s, 1H), 3.21-3.03 (m, 2H), 2.45-2.34 (m, 1H), 2.11-2.02 (m, 1H), 1.88-1.12 (m, 26H), 1.02-0.73 (m, 18H); ES Mass: [M+l] ⁺ 626.44.

Step 3: Synthesis of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)- 5a,5b,8,8 la^entamethyl-3a-(((S)-l-(5-phenyl-lH-imidazol-2-yl)ethyl)ca rbamoyl)-l-(prop- l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-dimethylcyclobutane-l,3- dicarboxylate:

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy- 5a,5b,8,8,l la-pentamethyl-N-((S)-l-(5-phenyl-lH-imidazol-2-yl)ethyl)-l- (prop-l-en-2- yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide (step 2, 1.0 g, 1.59 mmol, 1.0 eq) in toluene (20 ml) was added DMAP (0.39 g, 3.19 mmol, 2.0 eq) and (IS, 3R)-3- (benzyloxycarbonyl)-2,2-dimethylcyclobutane carboxylic 2,4,6-trichlorobenzoic anhydride (prepared as described in WO 2013/160810 A2, 1.4 g, 3.19 mmol, 2.0 eq). The reaction mixture was heated to 90°C for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was evaporated under reduced pressure and diluted with water. The aqueous layer was extracted with CH ₂ CI ₂ (2x100 ml). The combined organic layers were dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 50% EtOAc: n-Hexane as an eluent to obtain the desired product (1.0 g, yield: 71.9%) as a gummy syrup. 1H MR (300 MHz, CDC1 ₃ ): δ 12.18 (bs, 1H), 7.87-7.81 (m, 1H), 7.76-7.71 (m, 3H), 7.5 (s, 1H), 7.41-7.29 (m, 6H), 5.13-4.98 (m, 3H), 4.64 (s, 1H), 4.53 (s, 1H), 4.36-4.29 (m, 1H), 3.04-2.71 (m, 2H), 2.41-2.23 (m, 2H), 2.0-1.91 (m, 3H), 1.82-1.12 (m, 27H), 0.98-0.73 (m, 23H); ES Mass: [M+l] ⁺ 870.

Step 4: Synthesis of (lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR, 13bR)-5a,5b,8,8 la^entamethyl-3a-(((S)-l-(5-phenyl-lH-imidazol-2-yl)ethyl)ca rbamoyl)-l- (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy) carbonyl)cyclobutane-l- carboxylic acid:

To a stirred solution of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,

13bR)-5a,5b,8,8,l la-pentamethyl-3a-(((S)-l-(5-phenyl-lH-imidazol-2-yl)ethyl)c arbamoyl)- 1 -(prop- 1 -en-2-yl)icosahydro- 1 H-cyclopenta[a] chry sen-9-yl) ( 1 R,3 S )-2,2-dimethylcyclo butane- 1,3-dicarboxylate (step 3, 1.0 g, 1.15 mmol, 1.0 eq) in EtOAc: MeOH (1 : 1, 10 mL) was added ammonium formate (363 mg, 5.75, 5.0 eq), 10% Pd/C (100 mg) and stirred the reaction mixture for about 2 hours. The resultant mixture was filtered through celite and washed with methanol. The filtrate was concentrated, the obtained crude residue was dissolved in EtOAc (100 mL), washed with water, brine and organic layer was concentrated under reduced pressure to get the residue, which was purified by silicagel column chromatography by using 50% EtOAc: n-Hexane as an eluent to afford the desired compound (0.5 g, yield: 57.1%) as an off- white solid. 1H NMR (300 MHz, DMSO): δ 12.15 (s, 1H), 11.77 (s, 1H), 7.81-7.71 (m, 3H), 7.47 (bs, 1H), 7.38-7.28 (m, 2H), 7.18-7.11 (m, 1H), 5.08 (m, 1H), 4.64 (s, 1H), 4.53 (s, 1H), 4.36-4.29 (m, 1H), 3.08-2.98 (m, 1H), 2.83-2.7 (m, 1H), 2.62-2.43 (m, 1H), 2.32-2.18 (m, 2H), 2.01-1.72 (m, 3H), 1.68-1.21 (m, 25H), 1.01-0.73 (m, 23H); ES Mass: [M+l] ⁺ 780.6; HPLC purity: 90%.

Example 2: Preparation of 2.2-dimethyl-4-oxo-4-(((lR.3aS.5aR.5bRJaR.9S.l laR.l lbR. 13aR.13bR)-5a.5b.8.8.11a-pentamethyl-3a-(((S)-l-(5-phenyl-lH -imidazol-2-vnethvn carbamoyl)- 1 -(prop- l-en-2-yl)icosahvdro-lH-cvclopentara1chrvsen-9-yl)oxy)butano ic acid

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy- 5a,5b,8,8,l la-pentamethyl-N-((S) -(5-phenyl H-imidazol-2-yl)ethyl)-l-(prop-l-en-2- yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide (step 2, Example 1, 1.0 g, 1.59 mmol, 1.0 eq) and 2,2-dimethyl succinicanhydride (0.82 g, 6.39 mmol, 4.0 eq) in toluene (12 mL) was added DMAP (0.39 g, 3.19 mmol, 2.0 eq). The reaction mixture was heated at 90°C for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was concentrated under reduced pressure, cooled to 0°C, acidified to pH= 6-7 with IN HC1 and extracted with CH ₂ CI ₂ . The combined organic extracts were washed with water, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 50% EtOAc: n- Hexane as an eluent to afford the desired product (0.5 g, yield: 41.7%) as a brown color solid. 1H MR (300 MHz, DMSO): δ 12.16 (s, 1H), 11.75 (bs, 1H), 7.88 (bs, 1H), 7.73-7.68 (m, 2H), 7.53 (bs, 1H), 7.39-7.28 (m, 2H), 7.21 (bs, 1H), 5.05-5.0 (m, 1H), 4.64 (s, 1H), 4.52 (s, 1H), 4.39-4.31 (m, 1H), 3.03-2.93 (m, 1H), 2.29-2.18 (m, 1H), 1.99-1.9 (m, 1H), 1.82-1.02 (m, 31H), 1.02-0.73 (m, 20H); ES Mass: [M+l] ⁺ 794.64; HPLC purity: 84.0%.

Example 3: Preparation of 3.3-dimethyl-5-oxo-5-(((lR.3aS.5aR.5bRJaR.9S.l laR.l lbR. 13aR.13bR)-5a.5b.8.8.11a-pentamethyl-3a-(((S)-l-(5-phenyl-lH -imidazol-2-vnethvn carbamoyl)- 1 -(prop- l-en- -yl)icosahvdro-lH-cvclopentara1chrvsen-9-yl)oxy)pentanoic acid:

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy- 5a,5b,8,8,l la-pentamethyl-N-((S)-l-(5-phenyl-lH-imidazol-2-yl)ethyl)-l- (prop-l-en-2- yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide (Step 2, Example 1, 1.0 g, 1.59 mmol, 1.0 eq) and 3,3-dimethyl glutaric anhydride (0.91 g, 6.38 mmol, 4.0 eq) in toluene (12 mL) was added DMAP (0.39 g, 3.19 mmol, 2.0 eq). The reaction mixture was heated at 90°C for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was concentrated under reduced pressure, cooled to 0°C, acidified to P = 6-7 with IN HC1 and extracted with CH ₂ CI ₂ . The combined organic extracts were washed with water, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 30% EtOAc: n- Hexane as an eluent to afford the desired product (0.5 g, yield: 40.7%) as an off white solid. 1H MR (300 MHz, DMSO): δ 12.02 (s, 1H), 11.75 (bs, 1H), 7.9-7.83 (m, 1H), 7.78-7.71 (m, 2H), 7.6-7.52 (m, 1H), 7.41-7.29 (m, 2H), 7.24-7.18 (m, 1H), 5.05-5.01 (m, 1H), 4.64 (s, 1H), 4.53 (s, 1H), 4.41-4.34 (m, 1H), 3.03-2.95 (m, 1H), 2.4-2.34 (m, 2H), 2.32-2.21 (m, 3H), 1.98-1.89 (m, 1H), 1.82-1.13 (m, 22H), 1.12-0.73 (m, 27H); ES Mass: [M+l] ⁺ 768.63; HPLC purity: 90.1%.

Example-4: Preparation of (lR.3S)-3-((((lR.3aS.5aR.5bRJaR.9S. l laR. l lbR.13aR.13bR)- 3a-(((S)-l-(l-(2-(dimethylamino)ethyl)-4-phenyl-lH-imidazol- 2-yl)ethyl)carbamoyl)- 5a,5b,8,8J la-pentamethyl-l-(prop-l-en-2-yl)icosahvdro-lH-cvclopentara1 chrysen-9- yl)oxy)carbonyl)-2,2-dimethylcvclobutane- 1 -carboxylic acid

Step 1: Synthesis of 1 -benzyl 3-((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-3a-(((S)-l- (l-(2-(dimethylamino)ethyl)-4^henyl-lH-imidazol-2-yl)ethyl)c arbamoyl)-5a,5b,8,8,lla- pentamethyl-1 -(prop-1 -en-2-yl)icosahydro-lH-cyclopenta[a] chrysen-9-yl) (lR,3S)-2,2- dimethylcyclobutane-l,3-dicarboxylate:

To a stirred solution of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,

13bR)-5a,5b,8,8,l la-pentamethyl-3a-(((S)-l-(5-phenyl-lH-imidazol-2-yl)ethyl)c arbamoyl)- 1 -(prop- 1 -en-2-yl)icosahydro- 1 H-cyclopenta[a] chry sen-9-yl) ( 1 R,3 S )-2,2-dimethylcyclo butane- 1,3 -die arboxylate (step 3, Example 1, 0.6 g, 0.689 mmol, 1.0 eq) and 2-Chloro-N,N- dimethylethylamine hydrochloride (0.119 g, 0.83 mmol, 1.2 eq) in Ν,Ν-dimethyl formamide (6 mL) was added Cs ₂ C0 ₃ (0.67 g, 2.07 mmol, 3.0 eq). The reaction mixture was heated at 80°C for about 2 hours. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was diluted with water and extracted with EtOAc (2x100 ml). The combined organic extracts were dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The residue was purified by silicagel column chromatography by using 70% EtOAc: n-Hexane as an eluent to afford the desired product (0.35 g, yield: 53.8%) as an off white solid. 1H MR (300 MHz, DMSO): δ 8.01-7.98 (m, 1H), 7.73-7.68 (m, 2H), 7.61 (s, 1H), 7.41-7.29 (m, 7H), 7.19-7.11 (m, 1H), 5.12-5.03 (m, 2H), 4.65 (s, 1H), 4.52 (s, 1H), 4.36-4.29 (m, 1H), 4.08-3.83 (m, 2H), 3.08-2.79 (m, 4H), 2.28-2.13 (m, 9H), 2.01-1.88 (m, 2H), 1.83-1.12 (m, 30 H), 1.02-0.73 (m, 20H); ES Mass: [M+23] ⁺ 963.68.

Step 2: Synthesis of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,llaR,nbR,13aR,13bR)-3a-( ((S)- l-(l-(2-(dimethylamino)ethyl)-4^henyl-lH-imidazol-2-yl)ethyl )carbamoyl)-5a,5b,8,8,lla- pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chr ysen-9-yl)oxy)carbonyl)-2,2- dimethylcyclobutane-l -carboxylic acid:

To a stirred solution of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR) -3a-(((S)-l-(l-(2-(dimethylamino)ethyl)-4-phenyl-lH-imidazol -2-yl)ethyl)carbamoyl)- 5a,5b,8,8,l la-pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a] chrysen-9-yl) (lR,3S)-2,2-dimethylcyclobutane-l,3-dicarboxylate (step 1, 0.35 g, 0.372 mmol, 1.0 eq) in EtOAc: MeOH (1 : 1, 4 mL) added 10% Pd/C (50 mg). Hydrogen gas was bubbled through the reaction mixture via a balloon for about 2 hours. The resultant mixture was filtered through celite, washed with methanol and the filtrate was concentrated. The crude residue was purified by silicagel column chromatography by using 2% MeOH in CH ₂ C1 ₂ as an eluent to afford the desired compound (0.11 g, yield: 28.4%) as a white solid. 1H MR (300 MHz, DMSO): δ 12.19 (bs, 1H), 8.08-8.05 (m, 1H), 7.73-7.68 (m, 2H), 7.62 (s, 1H), 7.38-7.29 (m, 2H), 7.2-7.13 (m, 1H), 5.23-5.16 (m, 1H), 4.62 (s, 1H), 4.53 (s, 1H), 4.39-4.21 (m, 2H), 4.11- 3.98 (bs, 1H), 3.05-2.94 (m, 1H), 2.93-2.71 (m, 4H), 2.43-2.23 (m, 6H), 1.93-1.78 (m, 3H), 1.71-1.03 (m, 30H), 0.98-0.73 (m, 20H); ES Mass: [M+l] ⁺ 851.7; HPLC purity: 90.5%.

Example-5: Preparation of (lR,3S)-2,2-dimethyl-3-

((((lR.3aS.5aR.5bR JaR.9S.1 laR.1 IbR.13aR. 13bR)-5a.5b.8.8.1 la-pentamethyl-3a-((2-(5- phenyl-lH-imidazol-2-yl)propan-2-yl) carbamoyl)- 1 -(prop- 1 -en-2-yl)icosahydro- 1H- cyclopenta| ^" a ^" |chrysen-9-yl)oxy)carbonyl) cyclobutane-1 -carboxylic acid

Step 1: Synthesis of (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla- pentamethyl-3a-((2-(5-phenyl-lH-imidazol-2-yl)propan-2-yl)ca rbamoyl)-l-(prop-l-en-2- yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate:

To a stirred solution of 2-(5-phenyl-lH-imidazol-2-yl)propan-2-aminium chloride (Intermediate 3) and triethylamine (7.0 mL, 50.2 mmol, 5.0 eq) in CH ₂ CI ₂ (50 ml) at 0°C was added a solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-(chlorocarbonyl)- 5a,5b,8,8,l la-pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta [a]chrysen-9-yl acetate (prepared as described in WO 2013/160810 A2, 5.19 g, 10.04 mmol, 1.0 eq) in CH ₂ CI ₂ (50 ml). The reaction mixture was allowed to stir at room temperature for overnight. The reaction mixture was diluted with water and extracted with CH ₂ CI ₂ (3x200 ml). The combined organic extracts were dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The residue was purified by silicagel column chromatography by using 30% EtOAc: n-Hexane as an eluent to afford the desired product (2.0 g, yield: 29.4%) as a brown color solid. 1H MR (300 MHz, CDC1 ₃ ): δ 7.68-7.65 (m, 2H), 7.43-7.38 (m, 2H), 7.39-7.28 (m, 2H), 6.21 (s, 1H), 4.75 (s, 1H), 4.63 (s, 1H), 4.49-4.43 (m, 1H), 3.11-3.05 (m, 1H), 2.48- 2.39 (m, 1H), 2.03 (s, 3H), 1.95-1.11 (m, 27H), 1.02-0.68 (m, 21H); ES Mass: [M+l] ⁺ 682.6. Step 2: Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-9-hydroxy-5a,5b, 8,8, lla-pentamethyl-N-(2-(5-phenyl-lH-imidazol-2-yl)propan-2-yl) -l-(prop-l-en-2-yl) icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide:

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 5a,5b,8,8,l la-pentamethyl-3a-((2-(5-phenyl-lH-imidazol-2-yl)propan-2-yl )carbamoyl)-l- (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (step 1, 9.5 g, 13.92 mmol, 1.0 eq) in THF (80 ml), Methanol (80 ml) and water (100 mL) was added Lithium hydroxide mono hydrate (4.6 g, 111.36 mmol, 8.0 eq). The reaction mixture was stirred at room temperature for about 12 hours. TLC indicated starting material was consumed and the desired product was observed. The solvents were removed under reduced pressure, then the reaction mixture was diluted with water (200 mL) and extracted with ethyl acetate (2X250 mL). The combined organic layer was washed with brine, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude was purified by silicagel column chromatography by using 50% EtOAc: n-Hexane as an eluent to afford the desired product (6.0 g, yield: 67.3%) as an off-white solid. 1H MR (300 MHz, CDC1 ₃ ): δ 7.75-7.73 (m, 2H), 7.47-7.42 (m, 2H), 7.33-7.28 (m, 2H), 6.15 (s, 1H), 4.83 (s, 1H), 4.69 (s, 1H), 3.27-3.17 (m, 2H), 2.59- 2.48 (m, 1H), 2.01-1.13 (m, 30H), 1.08-0.72 (m, 18H); ES Mass: [M+l] ⁺ 662.57. Step 3: Synthesis of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)- 5a,5b,8,8,lla-pentamethyl-3a-((2-(5-phenyl-lH-imidazol-2-yl) propan-2-yl)carbamoyl)-l- (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-dimethylcyclobutane -1,3-dicarboxylate:

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy-

5a,5b,8,8,l la-pentamethyl-N-(2-(5-phenyl-lH-imidazol-2-yl)propan-2-yl)- l-(prop-l-en-2- yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide (step 2, 3.5 g, 5.47 mmol, 1.0 eq) in toluene (70 ml) was added DMAP (1.33 g, 10.95 mmol, 2.0 eq) and (IS, 3R)-3- (benzyloxycarbonyl)-2,2-dimethylcyclobutane carboxylic 2,4,6-trichlorobenzoic anhydride (prepared as described in WO 2013/160810 A2, 5.13 g, 10.95 mmol, 2.0 eq). The reaction mixture was heated to 90°C for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was evaporated under reduced pressure and diluted with water. The aqueous layer was extracted with CH ₂ C1 ₂ (2x100 ml). The combined organic layers were dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 50% EtOAc: n-Hexane as an eluent to obtain the desired product (4.0 g, yield: 82.82%) as an off- white solid. 1H MR (300 MHz, DMSO): δ 7.79-7.73 (m, 3H), 7.52 (s, 1H), 7.41-7.27 (m, 6H), 7.22-7.18 (m, 1H), 5.12-5.02 (m, 2H), 4.61 (s, 1H), 4.52 (s, 1H), 4.34-4.28 (m, 1H), 2.98-2.78 (m, 3H), 2.43-2.22 (m, 3H), 2.01-1.89 (m, 2H), 1.82-1.03 (m, 28H), 0.98-0.63 (m, 24H); ES Mass: [M+l] ⁺ 884.59.

Step 5: Synthesis of (lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR, 13bR)-5a,5b,8,8,lla-pentamethyl-3a-((2-(5-phenyl-lH-imidazol -2-yl)propan-2-yl) carbamoyl)-! -(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy )carbonyl) cyclobutane-1 -carboxylic acid:

To a stirred solution of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,

13bR)-5a,5b,8,8,l la-pentamethyl-3a-((2-(5-phenyl-lH-imidazol-2-yl)propan-2-yl ) carbamoyl)- l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2- dimethylcyclobutane-l,3-dicarboxylate (step 3, 4.0 g, 4.53 mmol, 1.0 eq) in EtOAc: MeOH (1 : 1, 40 mL) added 10% Pd/C (500 mg). Hydrogen gas was bubbled through the reaction mixture via a balloon for about 2 hours. The resultant mixture was filtered through celite, washed with methanol, the filtrate was concentrated and the crude residue was purified by silicagel column chromatography by using 50% EtOAc: n-Hexane as an eluent to afford the desired compound (1.2 g, yield: 33.4%) as a white solid. 1H MR (300 MHz, DMSO): δ 12.15 (s, 1H), 11.77 (s, 1H), 7.79-7.62 (m, 3H), 7.51 (m, 1H), 7.38-7.28 (m, 2H), 7.19-7.13 (m, 1H), 4.63 (s, 1H), 4.53 (s, 1H), 4.36-4.29 (m, 1H), 3.03-2.91 (m, 1H), 2.81-2.69 (m, 2H), 2.34-2.19 (m, 2H), 1.98-1.68 (m, 4H), 1.67-1.02 (m, 27H), 1.01-0.68 (m, 24H); ES Mass: [M+l] ⁺ 794.64; HPLC purity: 95%.

Example 6: Preparation of 2.2-dimethyl-4-oxo-4-(((lR.3aS.5aR.5bR.7aR.9S.l laR.l lbR. 13aR.13bR)-5a.5b.8.8.11a-pentamethyl-3a-((2-(5-phenyl-lH-imi dazol-2-vnpropan-2-vn carbamoyl)- 1 -(prop- l-en-2-yl)icosahydro-lH-cyclopenta[alchrysen-9-yl)oxy)butano ic acid

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy- 5a,5b,8,8,l la-pentamethyl-N-(2-(5-phenyl-lH-imidazol-2-yl)propan-2-yl)- l-(prop-l-en-2- yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide (Step 2, Example 5, 0.8 g, 1.25 mmol, 1.0 eq) and 2,2-dimethyl succinicanhydride (0.96 g, 7.5 mmol, 4.0 eq) in toluene (12 mL) was added DMAP (0.305 g, 2.5 mmol, 2.0 eq). The reaction mixture was heated at 90°C for overnight. TLC indicated starting material was consumed and the desired product was observed. The mixture was concentrated under reduced pressure, cooled to 0°C, acidified to pH= 6-7 with IN HC1 and extracted with CH ₂ CI ₂ . The combined organic extracts were washed with water, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 50% EtOAc: n- Hexane as an eluent to afford the desired product (0.55 g, yield: 57.3%) as a light yellow color solid. 1H MR (300 MHz, DMSO): δ 12.17 (s, 1H), 11.77 (bs, 1H), 11.78 (bs, 1H), 7.78-7.71 (m, 3H), 7.49 (bs, 1H), 7.35-7.29 (m, 2H), 7.18-7.16 (m, 1H), 4.63 (s, 1H), 4.52 (s, 1H), 4.38-4.29 (m, 1H), 2.99-2.97 (m, 1H), 2.45-2.42 (m, 2H), 2.28-2.21 (m, 1H), 1.97-1.91 (m, 1H), 1.83-1.02 (m, 30H), 1.0-0.68 (m, 21H); ES Mass: [M+l] ⁺ 768.7; HPLC purity: 82%. Example 7: Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((2-(l-(2-(dimethylamino)ethyl)-4-phenyl-lH-imidazol-2-yl )propan-2-yl)carbamoyl)- 5a,5b,8,8 J la-pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaral chrysen-9-yl) oxy)carbonyl)-2,2-dimethylcvclobutane- 1 -carboxylic acid

Step 1: Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-3a-((2-(l-(2- (dimethylamino)ethyl)-4^henyl-lH midazol-2-yl)propan-2-yl)carbamoyl)-5a,5b,8,8,lla- pentamethyl-1 -(prop-1 -en-2-yl)icosahydro-lH-cyclopenta[a] chrysen-9-yl acetate:

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 5a,5b,8,8,l la-pentamethyl-3a-((2-(5-phenyl-lH-imidazol-2-yl)propan-2-yl )carbamoyl)-l- (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (Step 1, Example 5 2.0 g, 2.93 mmol, 1.0 eq) and 2-Chloro-N,N-dimethylethylamine hydrochloride (0.506 g, 3.52 mmol, 1.2 eq) in Ν,Ν-dimethyl formamide (20 mL) was added Cs ₂ C0 ₃ (2.85 g, 8.79 mmol, 3.0 eq). The reaction mixture was heated at 80°C for about 2 hours. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was diluted with water and extracted with EtOAc (2x200 ml). The combined organic extracts were dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The residue was purified by silicagel column chromatography by using 70% EtOAc: n-Hexane as an eluent to afford the desired product (1.8 g, yield: 81.8%) as an off white solid. 1H NMR (300 MHz, DMSO): δ 7.86 (s, 1H), 7.73-7.71 (m, 2H), 7.59 (s, 1H), 7.36-7.3 (m, 2H), 7.18-7.13 (m, 1H), 4.6 (s, 1H), 4.51 (s, 1H), 4.38-4.33 (m, 1H), 4.21-4.11 (m, 2H), 2.92-2.83 (m, 1H), 2.68-2.59 (m, 2H), 2.21 (s, 6H), 1.99 (s, 3H), 1.99-1.88 (m, 1H), 1.73-1.06 (m, 28H), 0.98-0.73 (m, 19H); ES Mass: [M+l] ⁺ 753.52.

Step 2: Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,nbR,13aR,13bR)-N-(2-(l-(2- (dimethylamino)ethyl)-4^henyl-lH-imidazol-2-yl)propan-2-yl)- 9-hydroxy-5a,5b,8,8,lla- pentamethyl-l-(prop-l-en-2-yl)icosahydro-3aH-cyclopenta[a]ch rysene-3a-carboxamide:

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((2-(l- (2-(dimethylamino)ethyl)-4-phenyl-lH-imidazol-2-yl)propan-2- yl)carbamoyl)-5a,5b,8,8,l la- pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chr ysen-9-yl acetate (step 1, 1.5 g, 1.9 mmol, 1.0 eq) in THF (15 ml), methanol (15 ml) and water (20 ml) was added Lithium hydroxide mono hydrate (0.669 g, 15.94 mmol, 8.0 eq). The reaction mixture was stirred at room temperature for about 12 hours. TLC indicated starting material was consumed and the desired product was observed. The solvents were removed under reduced pressure, then the reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (2X100 mL). The combined organic layer was washed with brine and dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude was purified by silicagel column chromatography by using 5% MeOH in CH ₂ C1 ₂ as an eluent to afford the desired product (1.0 g, yield: 71%) as a light yellow solid. 1H NMR (300 MHz, DMSO): δ 7.86 (s, 1H), 7.73-7.69 (m, 2H), 7.62 (s, 1H), 7.39-7.3 (m, 2H), 7.21-7.13 (m, 1H), 4.6 (s, 1H), 4.49 (s, 1H), 4.28- 4.12 (m, 2H), 4.05-3.94 (m, 1H), 2.83-3.01 (m, 2H), 2.68-2.59 (m, 1H), 2.36-2.23 (m, 1H), 2.21 (s, 6H), 2.01-1.88 (m, 1H), 1.74-1.18 (m, 29H), 0.99-0.78 (m, 12H), 0.73 (s, 3H), 0.62 (s, 3H); ES Mass: [M+l] ⁺ 711.61. Step 3: Synthesis of 1 -benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-3a-((2-(l- (2-(dimethylamino)ethyl)-4-phenyl-lH-imidazol-2-yl)propan-2- yl)carbamoyl)-5a,5b,8,8,lla- pentamethyl-1 -(prop-1 -en-2-yl)icosahydro-lH-cyclopenta[a] chrysen-9-yl) (lR,3S)-2,2- dimethylcyclobutane-l,3-dicarboxylate:

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-N-(2-(l-(2- (dimethylamino)ethyl)-4-phenyl- lH-imidazol-2-yl)propan-2-yl)-9-hydroxy-5a,5b,8,8, 1 la- pentamethyl-l-(prop-l-en-2-yl)icosahydro-3aH-cyclopenta[a]ch rysene-3a-carboxamide (step 2, 1.0 g, 1.4 mmol, 1.0 eq) in toluene (70 ml) was added DMAP (0.343 g, 2.81 mmol, 2.0 eq) and (lS,3R)-3-(benzyloxycarbonyl)-2,2-dimethylcyclobutane carboxylic 2,4,6-trichloro benzoic anhydride (prepared as described in WO 2013/160810 A2, 1.32 g, 2.81 mmol, 2.0 eq). The reaction mixture was heated to 90°C for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was evaporated under reduced pressure and diluted with water. The aqueous layer was extracted with CH ₂ CI ₂ (2x100 ml). The combined organic layers were dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 70% EtOAc: n-Hexane as an eluent to obtain the desired product (0.8 g, yield: 60%) as an off-white solid. ¹ H MR (300 MHz, DMSO): δ 7.73-7.68 (m, 4H), 7.41-7.29 (m, 6H), 7.21-7.14 (m, 1H), 5.13-5.02 (m, 2H), 4.61 (s, 1H), 4.5 (s, 1H), 4.34-4.29 (m, 1H), 4.23-4.11 (m, 2H), 2.96-2.79 (m, 3H), 2.72-2.58 (m, 1H), 2.39-2.22 (m, 3H), 2.21 (s, 3H), 2.01-1.89 (m, 2H), 1.73-1.2 (m, 28H), 1.11-1.02 (m, 6H), 0.98-0.73 (m, 23H); ES Mass: [M+l] ⁺ 955.69.

Step 4: Synthesis of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-3a- ((2-(l- (2-(dimethylamino)ethyl)-4-phenyl-lH-imidazol-2-yl)propan-2- yl)carbamoyl)-5a,5b,8,8,lla- pentamethyl-1 -(prop-1 -en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbony l)-2,2- dimethylcyclobutane-1 -carboxylic acid:

To a stirred solution of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR) -3a-((2-(l-(2-(dimethylamino)ethyl)-4-phenyl-lH-imidazol-2-y l)propan-2-yl)carbamoyl)- 5a,5b,8,8,l la-pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a] chrysen-9-yl) (lR,3S)-2,2-dimethylcyclobutane-l,3-dicarboxylate (step 3, 0.8 g, 0.84 mmol, 1.0 eq) in EtOAc: MeOH (1 : 1, 6 mL) added 10% Pd/C (50 mg). Hydrogen gas was bubbled through the reaction mixture via a balloon for about 2 hours. The resultant mixture was filtered through celite, washed with methanol and the filtrate was concentrated. The crude residue was purified by silicagel column chromatography by using 2% MeOH in CH ₂ CI ₂ as an eluent to afford the desired compound (0.46 g, yield: 63.5%) as a white solid. 1H MR (300 MHz, CDCI ₃ ): δ 10.82 (bs, 1H), 7.71-7.48 (m, 3H), 7.4-7.32 (m, 2H), 7.19 (s, 1H), 6.04 (s, 1H), 4.76 (s, 1H), 4.62 (s, 1H), 4.48-4.39 (m, 1H), 3.16-3.06 (m, 1H), 2.86-2.74 (m, 2H), 2.63- 2.42 (m, 3H), 2.1-1.99 (m, 2H), 1.93-1.81 (m, 8H), 1.78-1.22 (m, 30H), 1.13-1.11 (m, 2H), 1.06 (s, 3H), 0.95 (s, 3H), 0.9-0.73 (m, 15H); ES Mass: [M+l] ⁺ 865.7; HPLC purity: 92.6%. Example 8: Preparation of 4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((2-(l- (2-(dimethylamino)ethyl)-4-phenyl-lH-imidazol-2-yl)propan-2- yl)carbamoyl)-5a,5b,8,8 J la- pentamethyl-l-(prop-l-en-2-yl)icosahvdro-lH-cvclopentaralchr vsen-9-yl)oxy)-2,2-dimethyl- 4-oxobutanoic acid

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-N-(2-(l-(2-

(dimethylamino)ethyl)-4-phenyl- lH-imidazol-2-yl)propan-2-yl)-9-hydroxy-5a,5b,8,8, 1 la- pentamethyl-l-(prop-l-en-2-yl)icosahydro-3aH-cyclopenta[a]ch rysene-3a-carboxamide (step 2, Example 7, 0.6 g, 0.8 mmol, 1.0 eq) and 2,2-dimethyl succinicanhydride (0.414 g, 3.2 mmol, 4.0 eq) in toluene (6 mL) was added DMAP (0.197 g, 1.61 mmol, 2.0 eq). The reaction mixture was heated at 90°C for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was concentrated under reduced pressure, cooled to 0°C, acidified to pH= 6-7 with IN HC1 and extracted with CH ₂ CI ₂ . The combined organic extracts were washed with water, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 5% MeOH in CH ₂ CI ₂ as an eluent to afford the desired product (0.3 g, yield: 42.4%) as an off white solid. 1H NMR (300 MHz, DMSO): δ 12.15 (bs, 1H), 7.87 (s, 1H), 7.73-7.68 (m, 2H), 7.59 (s, 1H), 7.38-7.29 (m, 2H), 7.18-7.13 (m, 1H), 4.61 (s, 1H), 4.5 (s, 1H), 4.39-4.32 (m, 1H), 4.22-4.12 (m, 1H), 3.05-2.93 (m, 1H), 2.91-2.82 (m, 1H), 2.7-2.58 (m, 2H), 2.32-2.16 (m, 2H), 2.21 (s, 6H), 2.01-1.9 (m, 1H), 1.73-1.01 (m, 29H), 1.01-0.73 (m, 24H); ES Mass: [M+l] ⁺ 839.58; HPLC purity: 88%. Example 9: Preparation of (lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR, 13aR,13bR)-5a,5b,8,8,l la-pentamethyl-3a-((2-(4-phenyl-l-(2-(pyrrolidin-l-yl)ethyl) -lH- imidazol-2-yl)propan-2-yl)carbamoyl)-l-(prop-l-en-2-yl)icosa hydro-lH-cyclopentaral chrvsen-9-yl)oxy)carbonyl)cvclobutane-l-carboxylic acid

Step 1: Synthesis of (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla- pentamethyl-3a-((2-(4-phenyl-l-(2-(pyrrolidin-l-yl)ethyl)-lH -imidazol-2-yl)propan-2- yl)carbamoyl)-l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]c hrysen-9-yl acetate:

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,l la-pentamethyl-3a-((2-(5-phenyl-lH-imidazol-2-yl)propan-2-yl )carbamoyl)-l-(prop-l- en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (Step 1, Example 5, 4.0 g, 5.86 mmol, 1.0 eq) and l-(2-Chloro ethylene)pyrrolidine hydrochloride (1.5 g, 8.79 mmol, 1.5 eq) in Ν,Ν-dimethyl formamide (40 mL) was added Cs ₂ C0 ₃ (5.72 g, 17.58 mmol, 3.0 eq). The reaction mixture was heated at 80°C for about 2 hours. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was diluted with water and extracted with EtOAc (2x200 ml). The combined organic extracts were dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The residue was purified by silicagel column chromatography by using 50% EtOAc: n-Hexane as an eluent to afford the desired product (1.8 g, yield: 39.4%) as a brown color solid. 1H MR (300 MHz, DMSO): δ 7.85 (s, 1H), 7.76-7.68 (m, 2H), 7.6 (s, 1H), 7.37-7.29 (m, 2H), 7.18-7.12 (m, 1H), 4.61 (s, 1H), 4.51 (s, 1H), 4.36-3.98 (m, 4H), 2.99-2.61 (m, 5H), 2.34-2.24 (m, 1H), 2.01 (s, 3H), 1.92-1.22 (m, 28H), 0.94-0.61 (m, 24H); ES Mass: [M+l] ⁺ 779.6.

Step 2: Synthesis of (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-9-hydroxy-

5a,5b,8,8,lla-pentamethyl-N-(2-(4-phenyl-l-(2-(pyrrolidin -l-yl)ethyl)-lH-imidazol-2- yl)propan-2-yl)-l-(prop-l-en-2-yl)icosahydro-3aH-cyclopenta[ a]chrysene-3a-carboxamide:

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 5a,5b,8,8,l la-pentamethyl-3a-((2-(4-phenyl -(2-(pyrrolidin -yl)ethyl)-lH-imidazol-2- yl)propan-2-yl)carbamoyl)-l-(prop-l-en-2-yl)icosahydro-lH-cy clopenta[a]chrysen-9-yl acetate (step 1, 1.8 g, 2.31 mmol, 1.0 eq) in THF (10 ml), methanol (10 ml) and water (10 mL) was added Lithium hydroxide mono hydrate (0.194 g, 4.62 mmol, 2.0 eq). The reaction mixture was stirred at room temperature for about 12 hours. TLC indicated starting material was consumed and the desired product was observed. The solvents were removed under reduced pressure, then the reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (2X100 mL). The combined organic layer was washed with brine, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude was purified by silicagel column chromatography by using 4% MeOH in CH ₂ C1 ₂ as an eluent to afford the desired product (0.9 g, yield: 53%) as a brown color solid. 1H MR (300 MHz, DMSO): δ 7.85 (s, 1H), 7.76-7.68 (m, 2H), 7.6 (s, 1H), 7.37-7.29 (m, 2H), 7.18-7.12 (m, 1H), 4.61 (s, 1H), 4.51 (s, 1H), 4.36-3.98 (m, 3H), 2.99-2.61 (m, 6H), 2.34-2.24 (m, 1H), 1.92-1.22 (m, 29H), 0.94-0.61 (m, 24H); ES Mass: [M+l] ⁺ 737.62.

Step 3: Synthesis of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)- 5a,5b,8,8 la^entamethyl-3a-((2-(4^henyl-l-(2-(pyrrolidin-l-yl)ethyl)-l H-imidazol-2- yl)propan-2-yl)carbamoyl)-l-(prop-l-en-2-yl)icosahydro-lH-cy clopenta[a]chrysen-9-yl) ( lR,3S)-2,2-dimethylcyclobutane-l,3-dicarboxylate:

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy 5a,5b,8,8,l la-pentamethyl-N-(2-(4-phenyl-l-(2-(pyrrolidin-l-yl)ethyl)-l H-imidazol-2- yl)propan-2-yl)-l-(prop-l-en-2-yl)icosahydro-3aH-cyclopenta[ a]chrysene-3a-carboxamide (step 2, 0.6 g, 0.814 mmol, 1.0 eq) in CH ₂ C1 ₂ (6.0 ml) was added (lS,3R)-3 (benzyloxycarbonyl)-2,2-dimethyl cyclobutanecarboxylic acid (prepared as described in WO

2013/160810 A2, 0.32 g, 1.22 mmol, 1.5 eq), DMAP (99 mg, 0.81 mmol, 1.0 eq) and DCC (0.252 g, 1.22 mmol, 1.5 eq) at 0°C. The reaction mixture was stirred for about 12 hours at room temperature. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was filtered through celite and the collected solid was washed with CH ₂ CI ₂ (10 mL). The filtrate was washed with water, the aqueous layer was extracted with CH ₂ CI ₂ (2x50 ml). The combined organic layers were dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 4% MeOH in CH ₂ CI ₂ as an eluent to obtain the desired product (0.6 g, yield: 75%) as an off white solid. 1H MR (300 MHz, DMSO): δ 7.85 (s, 1H), 7.76-7.68 (m, 2H), 7.6 (s, 1H), 7.37-7.29 (m, 8H), 7.18-7.12 (m, 1H), 5.13-5.01 (m, 2H), 4.61 (s, 1H), 4.51 (s, 1H), 4.36-3.98 (m, 4H), 2.91-2.69 (m, 5H), 2.34-2.24 (m, 2H), 2.01- 1.22 (m, 32H), 1.18-0.75 (m, 29H); ES Mass: [M+l] ⁺ 981.72.

Step 4: Synthesis of (lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR, 13bR)-5a,5b,8,8 la^entamethyl-3a-((2-(4^henyl-l-(2-(pyrrolidin-l-yl)ethyl)-l H-imidazol- 2-yl)propan-2-yl)carbamoyl)-l-(prop-l-en-2-yl)icosahydro-lH- cyclopenta[a]chrysen-9- yl)oxy )carbonyl)cyclobutane-l -carboxylic acid:

To a stirred solution of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR, 13bR)-5a,5b,8,8,l la-pentamethyl-3a-((2-(4-phenyl-l-(2-(pyrrolidin-l-yl)ethyl) -lH-imidazol- 2-yl)propan-2-yl)carbamoyl)-l-(prop-l-en-2-yl)icosahydro-lH- cyclopenta[a]chrysen-9-yl)

(lR,3S)-2,2-dimethylcyclobutane-l,3-dicarboxylate (step 3, 0.6 g, 0.61 mmol, 1.0 eq) in CH ₂ C1 ₂ : MeOH (1: 1, 12 mL) added ammonium formate (0.193 g, 3.06 mmol) and 10% Pd-C (65 mg, 0.06, 0.1 eq) at room temperature and stirred for about 2 hours. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was filtered through celite and the collected solid was washed with CH ₂ CI ₂ : MeOH (1: 1, 20 mL). The filtrate was concentrated under reduced pressure to afford the residue. The residue was re-dissolved in CH ₂ CI ₂ (20 mL), diluted with water. The organic layer was separated and the aqueous layer was extracted with CH ₂ CI ₂ (2x20 ml). The combined organic layers were dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 5% MeOH in CH ₂ CI ₂ as an eluent to obtain the desired product (0.2 g, yield: 36.7%) as an off- white solid. 1H NMR (300 MHz, DMSO): δ 12.14 (bs, 1H), 7.85 (s, 1H), 7.76-7.68 (m, 2H), 7.6 (s, 1H), 7.37-7.29 (m, 2H), 7.18-7.12 (m, 1H), 4.61 (s, 1H), 4.51 (s, 1H), 4.36-3.98 (m, 4H), 2.91-2.69 (m, 5H), 2.34- 2.24 (m, 2H), 2.01-1.22 (m, 32H), 1.18-0.75 (m, 29H); ES Mass: [M+l] ⁺ 891.7; HPLC purity: 93%. Example- 10: Preparation of (lR.3S)-2.2-dimethyl-3-

((((lR.3aS.5aR.5bR.7aR.9S.1 laR.1 IbR.13aR. 13bR)-5a.5b.8.8.1 la-pentamethyl-3a-(((S)-2- m ethyl- 1 -(5 -phenyl- lH-imidazol-2-yl)propyl) carbamoyl)- 1 -(prop- 1 -en-2-yl)icosahydro- 1H- cvclopentara1chrvsen-9-yl)oxy)carbonyl) cyclobutane-l-carboxylic acid

Step 1: Synthesis of (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla- pentamethyl-3a-(((S)-2-methyl-l-(5-phenyl-lH-imidazol-2-yl)p ropyl)carbamoyl)-l-(prop-l- en-2-yl)icosahydro-l H-cyclopenta[a] chrysen-9-yl acetate:

To a stirred solution of (S)-2-methyl-l-(5-phenyl-lH-imidazol-2-yl)propan-l- aminium chloride (Intermediate 4, 5.12 g, 23.9 mmol, 1.2 eq) and triethylamine (14.5 mL, 59.7 mmol, 5.0 eq) in CH ₂ CI ₂ (50 ml) at 0°C was added a solution of (lR,3aS,5aR,5bR,7aR,9S,l laR, HbR,13aR,13bR)-3a-(chlorocarbonyl)-5a,5b,8,8,l la- pentamethyl-l-(prop-l-en-2-yl) icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (prepared as described in WO 2013/160810 A2, 10.3 g, 19.9 mmol, 1.0 eq) in CH ₂ C1 ₂ (50 ml). The reaction mixture was allowed to stir at room temperature for overnight. After complete conversion of starting material, the reaction mixture was diluted with water and extracted with CH ₂ CI ₂ (3x200 ml). The combined organic extracts were dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The residue was purified by silicagel column chromatography by using 20% EtOAc: n-Hexane as an eluent to afford the desired product (10.0 g, yield: 72.5%) as a brown color solid. 1H NMR (300 MHz, DMSO): δ 11.74 (bs, 1H), 7.69-7.76 (m, 1H), 7.61-7.53 (m, 1H), 7.48 (s, 1H), 7.34-7.27 (m, 2H), 7.19-7.12 (m, 1H), 4.76-4.68 (m, 1H), 4.63 (s, 1H), 4.52 (s, 1H), 4.41-4.33 (m, 1H), 3.01-2.92 (m, 1H), 2.63- 2.54 (m, 1H), 2.31-2.13 (m, 4H), 1.98 (s, 3H), 1.92-1.68 (m, 3H), 1.67-1.03 (m, 19H), 0.96- 0.76 (m, 22H); ES Mass: [M+l] ⁺ 696.6. Step 2: Synthesis of (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-9-hydroxy- 5a,5b,8,8,lla-pentamethyl-N-((S)-2-methyl-l-(5-phenyl-lH-imi dazol-2-yl)propyl)-l-(prop-l- en-2-yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide:

stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-

5a,5b, 8,8,1 la-pentamethyl-3a-(((S)-2-methyl- l-(5-phenyl- lH-imidazol-2- yl)propyl)carbamoyl)-l-(prop-l-en-2-yl)icosahydro-lH-cyclope nta[a]chrysen-9-yl acetate (step 1, 15.0 g, 21.5 mmol, 1.0 eq) in THF (150 ml) and Methanol (150 ml) was added potassium carbonate (20.8 g, 150.0 mmol, 7.0 eq). The reaction mixture was stirred at room temperature for about 48 hours. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was filtered through a pad of celite and washed with CH ₂ CI ₂ . The filtrate was evaporated under reduced pressure and the crude was purified by silicagel column chromatography by using 30% EtOAc: n-Hexane as an eluent to afford the desired product (11 g, yield: 78.6%) as a light yellow solid. 1H MR (300 MHz, DMSO): δ 11.78 (bs, 1H), 7.78-7.69 (m, 1H), 7.63-7.54 (m, 1H), 7.49 (s, 1H), 7.38-7.21 (m, 2H), 7.19-7.11 (m, 1H), 4.77-4.69 (m, 1H), 4.62 (s, 1H), 4.52 (s, 1H), 4.27 (d, J = 5.1Hz, 1H),

3.01- 2.92 (m, 2H), 2.63-2.54 (m, 1H), 2.32-2.12 (m, 2H), 1.9-1.78 (m, 1H), 1.64-1.06 (m, 22H), 0.96-0.73 (m, 20H), 0.64 (s, 4H); ES Mass: [M+l] ⁺ 654.62.

Step 3: Synthesis of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)- 5a,5b,8,8,lla-pentamethyl-3a-(((S)-2-methyl-l-(5-phenyl-lH-i midazol-2- yl)propyl)carbamoyl)-l-(prop-l-en-2-yl)icosahydro-lH-cyclope nta[a]chrysen-9-yl) (1R,3S)-

2.2- dimethylcyclobutane- -dicarboxylate:

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy- 5a,5b,8,8,l la-pentamethyl-N-((S)-2-methyl-l-(5-phenyl-lH-imidazol-2-yl) propyl)-l-(prop- l-en-2-yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamid e (step 2, 1.5 g, 2.29 mmol, 1.0 eq) in toluene: pyridine (1: 1, 45 ml) was added DMAP (0.6 g, 4.59 mmol, 2.0 eq) and

(IS, 3R)-3-(benzyloxycarbonyl)-2,2-dimethylcyclobutane carboxylic 2,4,6-trichlorobenzoic anhydride (prepared as described in WO 2013/160810 A2, 2.14 g, 4.59 mmol, 2.0 eq). The reaction mixture was heated to 90°C for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was evaporated under reduced pressure and diluted with water. The aqueous layer was extracted with CH ₂ CI ₂ (2x50 ml). The combined organic layers were dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 30% EtOAc: n-Hexane as an eluent to afford the desired product (0.8 g, yield: 38.8%) as an off-white solid. 1H MR (300 MHz, DMSO): δ 11.74 (s, 1H), 7.73 (d, J = 7.5 Hz, 1H), 7.62-7.54 (m, 1H), 7.48 (s, 1H), 7.42-7.28 (m, 7H), 7.17-7.11 (m, 1H), 5.09 (ABq, J = 6.6 Hz, 2H), 4.77-4.68 (m, 1H), 4.63 (s, 1H), 4.51 (s, 1H), 4.36-4.29 (m, 1H), 2.98-2.77 (m, 2H), 2.39-2.13 (m, 3H), 1.98-1.82 (m, 2H), 1.63-1.07 (m, 29H), 0.97-0.73 (m, 26H); ES Mass: [M+l] ⁺ 898.7.

Step 4: Synthesis of (lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR, 13bR)-5a,5b,8,8,lla-pentamethyl-3a-(((S)-2-methyl-l-(5-pheny l-lH-imidazol-2-yl)propyl) carbamoyl)-! -(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy )carbonyl) cyclobutane-1 -carboxylic acid:

To a stirred solution of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR, 13bR)-5a,5b,8,8,l la-pentamethyl-3a-(((S)-2-methyl-l-(5-phenyl-lH-imidazol-2-y l)propyl) carbamoyl)- l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2- dimethylcyclobutane-l,3-dicarboxylate (step 3, 0.8 g, 0.89 mmol, 1.0 eq) in EtOAc: MeOH (1 : 1, 16 mL) added ammonium formate (281 mg, 4.45, 5.0 eq), 10% Pd/C (100 mg) and stirred the reaction mixture for about 2 hours. TLC indicated starting material was consumed and the desired product was observed. The resultant mixture was filtered through celite and washed with methanol. The collected filtrate was concentrated, obtained crude residue was dissolved in EtOAc (100 mL), washed with water, brine and the organic layer was concentrated under reduced pressure to get the residue, which was purified by silicagel column chromatography by using 30% EtOAc: n-Hexane as an eluent to afford the desired compound (0.16 g, yield: 22.2%) as a white solid. 1H NMR (300 MHz, DMSO): δ 12.17 (bs, 1H), 11.76 (s, 1H), 7.73 (d, J = 7.5 Hz, 1H), 7.6 (d, J = 8.1 Hz, 1H), 7.52-7.49 (m, 1H), 7.43- 7.24 (m, 2H), 7.17-7.12 (m, 1H), 4.77-4.68 (m, 1H), 4.61 (s, 1H), 4.51 (s, 1H), 3.02-2.89 (m, 1H), 2.82-2.71 (m, 1H), 2.64-2.53 (m, 1H), 2.34-2.14 (m, 2H), 1.91-1.83 (m, 2H), 1.67-1.05 (m, 29H), 0.98-0.71 (m, 26H); ES Mass: [M+l] ⁺ 808.55; HPLC purity: 90%. Example 11: Preparation of 2,2-dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR, 13aR,13bR)-5a,5b,8,8,l la-pentamethyl-3a-(((S)-2-methyl-l-(5-phenyl-lH-imidazol-2-y l) propyDcarbamoyl)- l-(prop- l-en-2-yl)icosahydro- lH-cyclopentaralchrysen-9-yl)oxy)butanoic acid

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy- 5a,5b,8,8,l la-pentamethyl-N-((S)-2-methyl-l-(5-phenyl-lH-imidazol-2-yl) propyl)-l-(prop- l-en-2-yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamid e (Step 2, Example 10, 1.5 g, 2.29 mmol, 1.0 eq) and 2,2-dimethyl succinicanhydride (1.18 g, 9.18 mmol, 4.0 eq) in toluene (15 mL) was added DMAP (0.559 g, 4.58 mmol, 2.0 eq). The reaction mixture was heated at 90°C for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was concentrated under reduced pressure, cooled to 0°C, acidified to P ^H = 6-7 with IN HC1 and extracted with CH ₂ C1 ₂ . The combined organic extracts were washed with water, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 60%

EtOAc: n-Hexane as an eluent to afford the desired product (1.0 g, yield: 55.9%) as an off white solid. 1H MR (300 MHz, DMSO): δ 12.1 (bs, 1H), 11.76 (bs, 1H), 7.75-7.68 (m, 2H), 7.59 (d, J = 9.0 Hz, 1H), 7.51-7.42 (m, 1H), 7.38-7.29 (m, 2H), 7.21-7.13 (m, 1H), 4.79-4.69 (m, 1H), 4.62 (s, 1H), 4.51 (s, 1H), 4.39-4.32 (m, 1H), 3.02-2.89 (m, 1H), 2.62-2.39 (m, 3H), 2.32-2.12 (m, 2H), 1.91-1.79 (m, 1H), 1.63-1.03 (m, 28H), 1.0-0.72 (m, 24H); ES Mass: [M+l] ⁺ 782.71; HPLC purity: 88.34%+9.82%.

Example 12: Preparation of (lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR, 13aR,13bR)-5a,5b,8,8,l la-pentamethyl-l-(l-methylcvclopropyl)-3a-((2-(5-phenyl-lH- imidazol-2-yl)propan-2-yl)carbamoyl)icosahvdro-lH-cvclopenta ralchrvsen-9-yl)oxy) carbonvPc vclobutane- 1 -carboxylic acid

Step 1: Synthesis of (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla- pentamethyl-l-(l-methylcyclopropyl)-3a-((2-(5-phenyl-lH-imid azol-2-yl)propan-2-yl) carbamoyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate:

stirred solution of 2-(5-phenyl-lH-imidazol-2-yl)propan-2-aminium chloride

(Intermediate 3, 3.0 g, 12.65 mmol, 1.2 eq) and triethylamine (7.3 mL, 52.7 mmol, 5.0 eq) in CH ₂ C1 ₂ (50 ml) at 0°C was added a solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR, 13aR,13bR)-3a-(chlorocarbonyl)-5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclopropyl) icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (prepared as described in WO 2013/160810 A2, 5.6 g, 10.54 mmol, 1.0 eq) in CH ₂ C1 ₂ (50 ml). The reaction mixture was allowed to stir at room temperature for overnight. The reaction mixture was diluted with water and extracted with CH ₂ C1 ₂ (3x200 ml). The combined organic extracts were dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The residue was purified by silicagel column chromatography by using 50% EtOAc: n-Hexane as an eluent to afford the desired product (5.0 g, yield: 68.2%) as a light yellow solid. 1H MR (300 MHz, CDC1 ₃ ): δ 11.56 (s, 1H), 7.76 (d, J = 7.2 Hz, 1H), 7.47-7.45 (m, 1H), 7.35 (t, J = 7.5Hz, 2H), 7.23-7.17 (m, 2H), 6.08 (s, 1H), 4.47 (t, J = 6.6 Hz, 1H), 2.39 (t, J = 9.6 Hz, 1H), 2.04 (s, 3H), 2.01 -1.85 (m, 3H), 1.8 (s, 6H), 1.72-1.53 (m, 6H), 1.5-1.02 (m, 12H), 0.96 (s, 3H), 0.91 (s, 3H), 0.85 (s, 3H), 0.82 (s, 9H), 0.66 (s, 3H), 0.5-0.32 (m, 2H), 0.31-0.21 (m, 2H); ES Mass: [M+l] ⁺ 696.5.

Step 2: Synthesis of (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-9-hydroxy- 5a,5b,8,8,lla-pentamethyl-l-(l-methylcyclopropyl)-N-(2-(5-ph enyl-lH-imidazol-2- yl)propan-2-yl)icosahydro- -cyclopenta[a]chrysene-3a-carboxamide:

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8, l la-pentamethyl-l-(l-methylcyclopropyl)-3a-((2-(5-phenyl-lH-i midazol-2-yl)propan-2-yl) carbamoyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (step 1, 2.5 g, 3.82 mmol, 1.0 eq) in THF (20 ml), Methanol (20 ml) and water (25 mL) was added Lithium hydroxide mono hydrate (1.3 g, 30.56 mmol, 8.0 eq). The reaction mixture was stirred at room temperature for about 12 hours. TLC indicated starting material was consumed and the desired product was observed. The solvents were removed under reduced pressure, then the reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (2X100 mL). The combined organic layer was washed with brine and dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude was purified by silicagel column chromatography by using 50% EtOAc: n-Hexane as an eluent to afford the desired product (1.8 g, yield: 76.6%) as an off-white solid. 1H NMR (300 MHz, DMSO): δ 11.79 (bs, 1H), 7.74 (d, J = 7.5Hz, 2H), 7.67 (s, 1H), 7.5 (s, 1H), 7.31 (t, J = 7.5Hz, 2H), 7.17-7.12 (m, 1H), 4.25 (d, J = 5.1 Hz, 1H), 2.5 (s, 6H), 2.38-2.21 (m, 3H), 1.91-1.62 (m, 4H), 1.62-1.58 (m, 6H), 1.56-1.02 (m, 13H), 0.92 (s, 3H), 0.91-0.83 (m, 6H), 0.76 (s, 3H), 0.7 (s, 3H), 0.62 (s, 3H), 0.35-0.24 (m, 2H), 0.21-0.11 (m, 2H); ES Mass: [M+Na] ⁺ 676.48.

Step 3: Synthesis of 1 -benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8, lla-pentamethyl-l-(l-methylcyclopropyl)-3a-((2-(5-phenyl-lH- imidazol-2-yl)propan-2- yl)carbamoyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-dimethylcyclobutane- 1,3-dicarboxylate:

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy- 5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclopropyl)-N-(2-(5-phenyl-lH-imi dazol-2-yl) propan-2-yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxam ide (step 2, 2.0 g, 3.06 mmol, 1.0 eq) in toluene (40 ml) was added DMAP (0.746 g, 6.12 mmol, 2.0 eq) and (IS, 3R)-3-(benzyloxycarbonyl)-2,2-dimethylcyclobutane carboxylic 2,4,6-trichlorobenzoic anhydride (prepared as described in WO 2013/160810 A2, 2.87 g, 6.12 mmol, 2.0 eq). The reaction mixture was heated to 90°C for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was evaporated under reduced pressure and diluted with water. The aqueous layer was extracted with CH ₂ C1 ₂ (2x100 ml). The combined organic layers were dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 50% EtOAc: n-Hexane as an eluent to obtain the desired product (1.6 g, yield: 58.2%) as an off-white solid. 1H NMR (300 MHz, DMSO): δ 12.12 (bs, 1H), 7.8-7.65 (m, 3H), 7.5 (s, 1H), 7.36-7.31 (m, 7H), 7.21-7.12 (m, 1H), 5.09 (q, J = 6.3 Hz, 2H), 4.32 (t, J = 7.5 Hz, 1H), 2.83-2.72 (m, 2H), 2.41-2.23 (m, 2H), 1.91-1.76 (m, 4H), 1.7-1.32 (m, 12H), 1.31-1.02 (m, 15H), 0.96-0.8 (m, 12H), 0.88-0.72 (m, 12H), 0.36-0.22 (m, 2H), 0.21-0.09 (m, 2H); ES Mass: [M+l] ⁺ 898.67.

Step 4: Synthesis of (lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR, 13bR)-5a,5b,8,8,lla-pentamethyl-l-(l-methylcyclopropyl)-3a-( (2-(5-phenyl-lH-imidazol-2- yl)propan-2-yl)carbamoyl)icosahydro-lH-cyclopenta[a]chrysen- 9-yl)oxy)carbonyl) cyclobutane-1 -carboxylic acid:

To a stirred solution of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,

13bR)-5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclopropyl)-3a-((2-(5-phenyl-lH-i midazol-2- yl)propan-2-yl)carbamoyl)icosahydro-lH-cyclopenta[a]chrysen- 9-yl) (lR,3S)-2,2-dimethyl cyclobutane-l,3-dicarboxylate (step 3, 1.6 g, 1.78 mmol, 1.0 eq) in EtOAc: MeOH (1 : 1, 32 mL) added 10% Pd/C (200 mg). Hydrogen gas was bubbled through the reaction mixture via a balloon for about 2 hours. The resultant reaction mixture was filtered through celite, washed with methanol, the filtrate was concentrated and the crude residue was purified by silicagel column chromatography by using 50% EtOAc: n-Hexane as an eluent to afford the desired compound (1.0 g, yield: 71.4%) as a white solid. 1H NMR (300 MHz, DMSO): δ 12.16 (s, 1H), 11.9 (s, 1H), 7.75-7.72 (m, 3H), 7.52 (s, 1H), 7.36-7.32 (m, 2H), 7.21-7.17 (m, 1H), 4.33 (t, J = 7.5 Hz, 1H), 2.82-2.74 (m, 2H), 2.41-2.23 (m, 2H), 1.91-1.78 (m, 4H), 1.69-1.38 (m, 11H), 1.37-1.06 (m, 16H), 0.98-0.81 (m, 12H), 0.87-0.73 (m, 12H), 0.36-0.23 (m, 2H), 0.22- 0.1 (m, 2H); ES Mass: [M+l] ⁺ 808.61; HPLC purity: 93%.

Example 13: Preparation of (lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR, 13aR,13bR)-5a,5b,8,8,l la-pentamethyl-3a-((l-(5-phenyl-lH-imidazol-2-yl)cvclobutyl) carbamoyl)- l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)ox y)carbonyl) cyclobutane-1 -carboxylic acid

Step 1: Synthesis of (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla- pentamethyl-3a-((l -(5-phenyl-lH-imidazol-2-yl)cyclobutyl)carbamoyl)-l -(prop-1 -en-2- yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate:

To a stirred solution of l-(5-phenyl-lH-imidazol-2-yl)cyclobutan-l -amine hydro chloride (Intermediate-4, 0.59 g, 2.3 mmol) in DCM (6 ml) and NEt ₃ (2.6 ml, 17.6 mmol), was added a solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a- (chlorocarbonyl)-5a,5b, 8,8,1 la-pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH- cyclopenta[a]chrysen-9-yl acetate (prepared as described in WO 2013/160810 A2, 1.2 g, 2.3 mmol) in DCM (12 ml) at 0°C and allowed to stir at room temperature for about 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM, washed with water, brine and dried over Na ₂ S0 ₄ . The solvent was evaporated and purified by silica gel column (elution 30% EtOAc in n-hexane) to afford the title compound (0.5 g, yield: 32%) as an off white solid. 1H MR (300 MHz, CDC1 ₃ ): δ 7.68 (d, J= 7.5 Hz, 2H), 7.41 (dd, J= 7.5 Hz, 7.5Hz, 2H), 7.32-7.23 (m, 2H), 4.72 (s, 1H), 4.61 (s, 1H), 4.47-4.42 (m, 1H), 3.12-2.92 (m, 3H), 2.46-1.93 (m, 6H), 2.04 (s, 3H), 1.92-1.05 (m, 26H), 0.94 (s, 3H), 0.81 (s, 3H), 0.80 (s, 3H), 0.75-0.70 (m, 6H); ES Mass: 694.51 [M+H] ⁺ . Step 2: Synthesis of (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-9-hydroxy- 5a,5b,8,8,lla-pentamethyl-N-(l-(5-phenyl-lH-imidazol-2-yl)cy clobutyl)-l-(prop-l-en-2- yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide:

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 5a,5b,8,8,l la-pentamethyl-3a-((l-(5-phenyl-lH-imidazol-2-yl)cyclobutyl) carbamoyl)-l-

(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (step 1, 0.5 g, 0.72 mmol) in MeOH: THF (5 ml: 5 ml), was added 4N NaOH (5 ml) at 0°C and allowed to stir at room temperature for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure. The reaction mixture was diluted with DCM, washed with water, brine and dried over Na ₂ S0 ₄ . The solvent was evaporated and the resulting solid was taken in hexane, stirred for about one hour and filtered to afford the title compound (0.34 g, yield: 73%) as a white solid. 1H NMR (300 MHz, CDC1 ₃ ): δ 7.68 (d, J = 7.5 Hz, 2H), 7.38 (dd, J = 7.5 Hz, 7.5Hz, 2H), 7.27-7.25 (m, 1H), 7.22 (s, 1H), 6.50 (s, 1H), 4.75 (s, 1H), 4.61 (s, 1H), 3.19-3.09 (m, 3H), 2.95-2.93 (m, 1H), 2.44-2.35 (m, 3H), 2.05-1.09 (m, 22H), 0.98-0.83 (m, 11H), 0.76- 0.70 (m, 9H), 0.68-0.61 (m, 1H); ES Mass: 652.46 [M+H] ⁺ .

Step 3: Synthesis of 1 -benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8, lla-pentamethyl-3a-((l-(5-phenyl-lH-imidazol-2-yl)cyclobutyl )carbamoyl)-l-(prop-l-en-2- yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-dimethylcyclobutane-l,3- dicarboxylate:

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy- 5a,5b,8,8, 11 a-pentamethyl-N-( 1 -(5-phenyl- 1 H-imidazol-2-yl)cyclobutyl)- 1 -(prop- 1 -en-2- yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide (step 2, 0.2 g, 0.3 mmol) and (lS,3R)-3-((benzyloxy)carbonyl)-2,2-dimethylcyclobutane-l-ca rboxylic acid (prepared as described in WO 2013/160810 A2, 0.12 g, 0.46 mmol) and DMAP (0.01 g, cat) in DCM (3 ml), was slowly added DCC (0.13 g, 0.61 mmol) in DCM (2 ml) at 0°C and allowed to stir at room temperature for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM, washed with water, saturated NaHC0 ₃ solution, brine and dried over Na ₂ S0 ₄ . The solvent was evaporated, to the resulting solid was added DCM (3 ml) and stirred for about 1 hour and filtered. The filtrate was concentrated under reduced pressure to afford the title compound (0.21 g, yield: 76%) as a white solid. 1H NMR (300 MHz, CDCI ₃ ): δ 7.28 (m, 10H), 7.14 (s, 1H), 6.19 (s, 1H), 5.05 (ABq, J= 12.3 Hz, 2H), 4.69 (s, 1H), 4.55 (s, 1H), 4.35-4.33 (m, 1H), 3.08-2.50 (m, 9H), 1.97-1.05 (m, 30H), 0.92- 0.83 (m, 9H), 0.73-0.64 (m, 12H).

Step 4: Synthesis of (lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR, 13bR)-5a,5b,8,8,lla-pentamethyl-3a-((l-(5-phenyl-lH-imidazol -2-yl)cyclobutyl)carbamoyl)- l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)ox y)carbonyl)cyclobutane-l- carboxylic acid:

To a stirred solution of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR) -5a,5b,8,8,l la-pentamethyl-3a-((l-(5-phenyl-lH-imidazol-2-yl)cyclobutyl) carbamoyl)-l- (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-dimethylcyclo butane- 1,3 -die arboxylate (step 3, 0.21 g, 0.23 mmol) in THF (5 ml), were added N(Et) ₃ (0.15 ml, 1 mmol), triethyl silane (0.2 ml, 0.75 mmol) and Pd(OAc) ₂ (0.02 g, cat). The reaction mixture was heated to 70°C and continued for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc, washed with water, saturated NaHC0 ₃ solution, brine and dried over Na ₂ S0 ₄ . The solvent was evaporated under reduced pressure and purified by silica gel column (elution 5% MeOH in DCM) to afford the title compound (0.08 g, yield: 42%) as a white solid. 1H NMR (300 MHz, CDC1 ₃ ): δ 7.75-7.71 (m, 2H), 7.47-7.38 (m, 4H), 4.67 (s, 1H), 4.58 (s, 1H), 4.45-4.41 (m, 1H), 3.13- 1.75 (m, 16H), 1.59-0.71 (m, 39H), 0.61-0.50 (m, 6H); ES Mass: 806.62 [M+H] ⁺ .

Example 14: Preparation of 2,2-dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR, 13aR,13bR)-5a,5b,8,8,l la-pentamethyl-3a-((l-(5-phenyl-lH-imidazol-2-yl)cvclobutyl) carbamoyl)- l-(prop-l-e - -yl)icosahvdro-lH-cvclopentaralchrvsen-9-yl)oxy)butanoic acid

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy-

5a,5b,8,8, 11 a-pentamethyl-N-( 1 -(5-phenyl- 1 H-imidazol-2-yl)cyclobutyl)- 1 -(prop- 1 -en-2- yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide (Step 2, Example 13, 0.13 g, 0.2 mmol) and 2,2-dimethyl succinicanhydride (0.10 g, 0.8 mmol) in toluene (7 ml), was added DMAP (0.05 g, 0.4 mmol). The reaction mixture was heated at 90°C for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure, cooled to 0°C, acidified to P = 6 with IN HC1 and extracted with DCM. The combined organic extracts were washed with water, brine, dried over Na ₂ S0 ₄ then the solvent was evaporated under reduced pressure and to the resulting solid was recrystallized from ACN to give the title compound (0.06 g, yield: 39%). 1H NMR (300 MHz, CDC1 ₃ ): δ 7.76-7.74 (m, 2H), 7.47-7.38 (m, 3H), 7.22 (s, 1H), 4.63 (s, 1H), 4.57 (s, 1H), 4.38-4.33 (m, 1H), 3.12-1.94 (m, 14H), 1.58-1.08 (m, 22H), 0.95-0.77 (m, 21H); ES Mass: 780.67 [M+H] ⁺ .

Example 15: Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((l-(5-(4-fluorophenyl)-lH-imidazol-2-yl)cvclobutyl)carba moyl)-5a,5b,8,8 J la-penta methyl- l-(prop-l-en-2-yl)icosahvdro-lH-cvclopentaralchrvsen-9-yl)ox y)carbonyl)-2,2- dimethylcyclobutane- 1 -carboxylic acid

Step 1: Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-3a-((l-(5-(4- fluorophenyl) -1 H-imidazol-2-yl)cyclobutyl)carbamoyl)-5a,5b, 8,8 ,1 la-pentamethyl-1 -(prop- l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate:

To a stirred solution of l-(5-(4-fluorophenyl)-lH-imidazol-2-yl)cyclobutan-l -amine hydro chloride (Intermediate 5, 0.64 g, 2.4 mmol) in DCM (8 ml) and NEt ₃ (2 ml, 13.5 mmol), was added a solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a- (chlorocarbonyl)-5a,5b,8,8, 1 la-pentamethyl- l-(prop- l-en-2-yl)icosahydro- 1H- cyclopenta[a]chrysen-9-yl acetate (prepared as described in WO 2013/160810 A2, 1 g, 1.94 mmol) in DCM (10 ml) at 0°C and allowed to stir at room temperature for about 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM, washed with water, brine, dried over Na ₂ S0 ₄ , and the solvent was evaporated and purified by silica gel column (elution 30% EtOAc in n- hexane) to afford the title compound (0.7 g, yield: 51%)as an off white solid. ¹ H NMR (300 MHz, CDC1 ₃ ): δ 10.48 (s, 1H), 7.78-7.71 (m, 1H), 7.15 (s, 1H), 7.08-7.03 (m, 2H), 4.76 (s, 1H), 4.62 (s, 1H), 4.48- 4.42 (m, 1H), 3.15-2.93 (m, 3H), 2.41-2.30 (m, 4H), 2.03 (s, 3H), 2.07-0.72 (m, 44H); ES Mass: 712.60 [M+H] ⁺ .

Step 2: Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-N-(l-(5-(4- fluorophenyl)-lH-imidazol-2-yl)cyclobutyl)-9-hydroxy-5a,5b,8 ,8,lla-pentamethyl-l-(prop-l- en-2-yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide:

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(5- (4-fluorophenyl)-lH-imidazol-2-yl)cyclobutyl)carbamoyl)-5a,5 b,8,8,l la-pentamethyl-l- (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (step 1, 0.7 g, 0.98 mmol) in MeOH: THF (7 ml: 7 ml), was added 4N NaOH (7 ml) at 0°C and allowed to stir at room temperature for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure. The reaction mixture was diluted with DCM, washed with water, brine and dried over Na ₂ S0 ₄ . The solvent was evaporated and the resulting solid was taken in hexane and stirred for about one hour and filtered to afford the title compound (0.6 g, yield: 91%) as a white solid. 1H MR (300 MHz, CDC1 ₃ ): δ 10.48 (s, 1H), 7.77-7.73 (m, 2H), 7.15 (s, 1H), 7.05 (dd, J= 8.4 Hz, 2H), 6.23 (s, 1H), 4.76 (s, 1H), 4.62 (s, 1H), 3.14-2.93 (m, 4H), 2.39-2.30 (m, 3H), 2.08-1.84 (m, 4H), 1.62-1.08 (m, 19H), 0.94-0.59 (m, 20H); ES Mass: 670.55 [M+H] ⁺ .

Step 3: Synthesis of 1 -benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-3a-((l-(5- ( 4 -fluorophenyl)-! H-imidazol-2-yl )cyclobutyl )carbamoyl)-5a, 5b, 8, 8, 1 la-pentamethyl-1 -

(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-dimethylcyclobutane 1,3-dicarboxylate:

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-N-(l-(5-(4- fluorophenyl)- lH-imidazol-2-yl)cyclobutyl)-9-hydroxy-5a,5b, 8,8,1 la-pentamethyl-l-(prop- l-en-2-yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamid e (step 2, 0.3 g, 0.44 mmol) and (lS,3R)-3-((benzyloxy)carbonyl)-2,2-dimethylcyclobutane-l-ca rboxylic acid (prepared as described in WO 2013/160810 A2, 0.18 g, 0.67 mmol) and DMAP (0.01 g, cat) in DCM (4 ml), was slowly added DCC (0.18 g, 0.89 mmol) in DCM (3 ml) at 0°C and allowed to stir at room temperature for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM, washed with water, saturated NaHC0 ₃ solution, brine and dried over Na ₂ S0 ₄ . The solvent was evaporated, to the resulting solid was added DCM (4 ml), stirred for about one hour and filtered. The filtrate was concentrated under reduced pressure to afford the title compound (0.4 g, yield: 97%) as a white solid. 1H MR (300 MHz, CDC1 ₃ ): δ 10.61 (brs, 1H), 7.80-7.61 (m, 2H), 7.34 (m, 5H), 7.15 (s, 1H), 7.08- 7.03 (m, 2H), 6.30 (s, 1H), 5.12 (m, 2H), 4.75 (s, 1H), 4.61 (s, 1H), 4.43 (m, 1H), 3.1 -1.82 (m, 20H), 1.69-1.12 (m, 21H), 0.95-0.81 (m, 18H); ES Mass: 914.6 [M+H] ⁺ .

Step 4: Synthesis of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-3a- ((l-(5- ( 4 -fluorophenyl)-! H-imidazol-2-yl )cyclobutyl )carbamoyl)-5a, 5b, 8, 8, 1 la-pentamethyl-1 - (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy) carbonyl)-2,2-dimethyl cyclobutane-1 -carboxylic acid:

To a stirred solution of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR) -3a-((l-(5-(4-fluorophenyl)-lH-imidazol-2-yl)cyclobutyl)carb amoyl)-5a,5b,8,8,l la-penta methyl- l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-dimethyl cyclobutane-l,3-dicarboxylate (step 3, 0.4 g, 0.43 mmol) in THF (5 ml), were added N(Et) ₃ (0.25 ml, 1.7 mmol), triethyl silane (0.22 ml, 1.4 mmol) and Pd(OAc) ₂ (0.04 g, cat). The reaction mixture was heated to 70°C and continued for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc, washed with water, saturated NaHC0 ₃ solution, brine and dried over Na ₂ S0 ₄ . The solvent was evaporated under reduced pressure and purified by silica gel column (elution 5% MeOH in DCM) to afford the title compound (0.14 g, yield: 40%) as a white solid. 1H NMR (300 MHz, DMSO- D ₆ ): δ 12.08 (brs, 1H), 11.46 (s, 1H), 8.13 (s, 1H), 7.76 (dd, J = 5.7 Hz, 8.4 Hz, 2H), 7.41 (s, 1H), 7.12 (dd, J = 8.4 Hz, 8.4 Hz, 2H), 4.60 (s, 1H), 4.50 (s, 1H), 4.34-4.29 (m, 1H), 2.98- 2.92 (m, 1H), 2.81-2.75 (m, 2H), 2.60-2.22 (m, 6H), 1.98-1.01 (m, 31H), 0.95-0.69 (m, 19H); ES Mass: 824.5 [M+H] ⁺ .

Example 16: Preparation of 4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(5- (4-fluorophenyl)-lH-imidazol-2-yl)cyclobutyl)carbamoyl)-5a.5 b.8.8.1 la-pentamethyl- 1- (prop-l-en-2-yl)icosahvdro-lH-cvclopentaralchrvsen-9-yl)oxy) -2.2-dimethyl-4-oxobutanoic acid

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-N-(l-(5-(4- fluorophenyl)-lH-imidazol-2-yl)cyclobutyl)-9-hydroxy-5a,5b,8 ,8,l la-pentamethyl-l-(prop- l-en-2-yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamid e (Step 2, Example 15, 0.15 g, 0.22 mmol) and 2,2-dimethyl succinicanhydride (0.11 g, 0.89 mmol) in toluene (5 ml), was added DMAP (0.055 g, 0.44 mmol). The reaction mixture was heated at 90°C for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure, cooled to 0°C, acidified to pH=6 with IN HCl and extracted with DCM. The combined organic extracts were washed with water, brine, dried over Na ₂ S0 ₄ then the solvent was evaporated under reduced pressure and the resulting solid was recrystallized from ACN to give the title compound (0.08 g, yield: 45%). 1H MR (300 MHz, DMSO-D ₆ ): δ 12.15 (s, 1H), 11.47 (s, 1H), 8.14 (s, 1H), 7.76 (dd, J = 6.0 Hz, 8.7 Hz, 2H), 7.41 (s, 1H), 7.13 (dd, J = 8.7 Hz, 8.7 Hz, 2H), 4.60 (s, 1H), 4.50 (s, 1H), 4.37-4.31 (m, 1H), 2.94-2.91 (m, 1H), 2.65-2.32 (m, 8H), 1.99-1.01 (m, 29H), 0.90-0.68 (m, 19H); ES Mass: 798.5 [M+H] ⁺ .

Example 17: Preparation of 4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(5- (4-fluorophenyl)-lH-imidazol-2-yl)cvclobutyl)carbamoyl)-5a,5 b,8,8 j la-pentamethyl-l-d- methylcyclopropyl)icosahydro-lH-cyclopentaralchrysen-9-yl)ox y)-2,2-dimethyl-4-oxo butanoic acid

Step 1: Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-3a-((l-(5-(4- fluorophenyl)-lH-imidazol-2-yl)cyclobutyl)carbamoyl)-5a,5b,8 ,8,lla-pentamethyl-l-(l- methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate:

To a stirred solution of l-(5-(4-fluorophenyl)-lH-imidazol-2-yl)cyclobutan-l -amine hydrochloride (Intermediate 5, 0.64 g, 2.4 mmol) in DCM (8 ml) and NEt ₃ (2 ml, 13.5 mmol), was added a solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-(chloro carbonyl)-5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclopropyl)icosahydro-lH-cyclopen ta[a] chrysen-9-yl acetate (prepared as described in WO 2013/160810, 1 g, 1.94 mmol) in DCM (10 ml) at 0°C and allowed to stir at room temperature for about 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM, washed with water, brine and dried over Na ₂ S0 ₄ . The solvent was evaporated and purified by silica gel column (elution 30% EtOAc in n- hexane) to afford the title compound (1 g, yield: 73%) as an off white solid. 1H MR (300 MHz, CDC1 ₃ ): δ 7.68-7.64 (m, 2H), 7.14 (s, 1H), 7.07 (dd, J = 8.7 Hz, 8.7 Hz, 2H), 6.51 (s, 1H), 4.49-4.44 (m, 1H), 3.12-3.04 (m, 1H), 2.96-2.84 (m, 1H), 2.41-2.20 (m, 4H), 2.05 (s, 3H), 1.94-1.05 (m, 24H), 1.01-0.71 (m, 20H), 0.44-0.34 (m, 2H), 0.28-0.21 (m, 2H).

Step 2: Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-N-(l-(5-(4- fluorophenyl)-lH midazol-2-yl)cyclobutyl)-9-hydroxy-5a,5b,8,8,lla-pentamethyl -l-(l- methylcyclopropyl)icosahydro-3aH-cyclopenta[a]chrysene-3a-ca rboxamide:

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(5- (4-fluorophenyl)-lH-imidazol-2-yl)cyclobutyl)carbamoyl)-5a,5 b, 8,8,1 la-pentamethyl- 1-(1- methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (step 1, 1 g, 1.3 mmol) in MeOH: THF (10 ml: 10 ml), was added 4N NaOH (10 ml) at 0°C and allowed to stir at room temperature for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure. The reaction mixture was diluted with DCM, washed with water, brine and dried over Na ₂ S0 ₄ . The solvent was evaporated, the resulting solid was taken in hexane and stirred for about one hour and filtered to afford the title compound (0.48 g, yield: 52%) as a white solid. 1H NMR (300 MHz, CDC1 ₃ ): δ 7.70-7.65 (m, 2H), 7.13-7.06 (m, 3H), 3.19-3.08 (m, 2H), 2.96-2.84 (m, 1H), 2.44- 2.23 (m, 3H), 2.04-1.94 (m, 4H), 1.77-1.08 (m, 22H), 1.02-0.65 (m, 20H), 0.38-0.33 (m, 2H), 0.26-0.21 (m, 2H); ES Mass: 684.53 [M+H] ⁺ .

Step 3: Synthesis of 4-(((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-3a-((l-(5-(4- fluorophenyl)-lH-imidazol-2-yl)cyclobutyl)carbamoyl)-5a,5b,8 ,8,lla-pentamethyl-l-(l- methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)ox y)-2,2-dimethyl-4-oxo butanoic acid:

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-N-(l-(5-(4- fluorophenyl)-lH-imidazol-2-yl)cyclobutyl)-9-hydroxy-5a,5b, 8,8,1 la-pentamethyl-l-(l- methylcyclopropyl)icosahydro-3aH-cyclopenta[a]chrysene-3a-ca rboxamide (step 2, 0.125 g, 0.18 mmol) and 2,2-dimethyl succinicanhydride (0.093 g, 0.73 mmol) in toluene (5 ml), was added DMAP (0.044 g, 0.36 mmol). The reaction mixture was heated at 90°C for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure, cooled to 0°C, acidified to pH=6 with IN HCl and extracted with DCM. The combined organic extracts was washed with water, brine, dried over Na ₂ S0 ₄ then the solvent was evaporated under reduced pressure and the resulting solid was recrystallized from ACN to give the title compound (0.066 g, yield: 45%). 1H NMR (300 MHz, DMSO-D ₆ ): δ 12.16 (s, 1H), 11.43 (s, 1H), 8.07 (s, 1H), 7.75 (m, 2H), 7.40 (s, 1H), 7.13 (dd, J = 8.4 Hz, 8.4 Hz, 2H), 4.38-4.33 (m, 1H), 2.62-2.35 (m, 8H), 1.92-1.15 (m, 30H), 0.91-0.67 (m, 19H), 0.31-0.23 (m, 2H), 0.20-0.10 (m, 2H); ES Mass: 812.6 [M+H] ⁺ .

Example 18: Preparation of 2,2-dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR, 13aRJ3bR)-5a,5b,8,8 J la-pentamethyl-3a-((l-(5-phenyl-lH-imidazol-2-yl)cyclopentyl ) carbamoyl)- l-(prop-l- -2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)butanoic acid

Step 1: Synthesis of (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla- pentamethyl-3a-((l -(5-phenyl-lH-imidazol-2-yl)cyclopentyl)carbamoyl)-l -(prop-1 -en-2- yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate:

To a stirred solution of l-(5-phenyl-lH-imidazol-2-yl)cyclopentan-l -amine hydro chloride (Intermediate 6, 0.16 g, 0.61 mmol) in DCM (3 ml) and NEt ₃ (0.5 ml, 3.4 mmol), was added a solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a- (chlorocarbonyl)-5a,5b, 8,8,1 la-pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH- cyclopenta[a]chrysen-9-yl acetate (prepared as described in WO 2013/160810 A2, 0.25 g, 0.49 mmol) in DCM (4 ml) at 0°C and allowed to stir at room temperature for about 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM, washed with water, brine and dried over Na ₂ S0 ₄ . The solvent was evaporated and purified by silica gel column (elution 30% EtOAc in n- hexane) to afford the title compound (0.1 g, yield: 30%) as an off white solid. 1H MR (300 MHz, CDC1 ₃ ): δ 10.89 (s, 1H), 7.75 (m, 2H), 7.54-7.47 (m, 1H), 7.38-7.33 (m, 2H), 7.19 (s, 1H), 5.89 (s, 1H), 4.76 (s, 1H), 4.62 (s, 1H), 4.44 (m, 1H), 3.10 (m, 1H), 2.70-2.18 (m, 5H), 2.04 (s, 3H), 1.86- 1.07 (m, 29H), 0.93-0.63 (m, 16H); ES Mass: 708.55 [M+H] ⁺ .

Step 2: Synthesis of (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-9-hydroxy- 5a,5b,8,8 la^entamethyl-N-(l-(5-phenyl-lH-imidazol-2-yl)cyclopentyl)-l -(prop-l-en-2- yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide:

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 5a,5b,8,8,l la-pentamethyl-3a-((l-(5-phenyl-lH-imidazol-2-yl)cyclopentyl )carbamoyl)-l-

(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (step 1, 0.1 g, 0.14 mmol) in MeOH: THF (2 ml: 2 ml), was added 4N NaOH (2 ml) at 0°C and allowed to stir at room temperature for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure. The reaction mixture was diluted with DCM, washed with water, brine and dried over Na ₂ S0 ₄ . The solvent was evaporated, the resulting solid was taken in hexane, stirred for about one hour and filtered to afford the title compound (0.06 g, yield: 65%) as a white solid. 1H NMR (300 MHz, CDC1 ₃ ): δ 10.89 (s, 1H), 7.54-7.51 (m, 2H), 7.38-7.33 (m, 1H), 7.29-7.28 (m, 2H), 7.18 (s, 1H), 5.89 (s, 1H), 4.76 (s, 1H), 4.61 (s, 1H), 3.15 (m, 1H), 2.70-1.69 (m, 20H), 1.47-0.60 (m, 32H).

Step 3: Synthesis of 2,2-dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR) -5a,5b,8,8,lla-pentamethyl-3a-((l-(5-phenyl-lH-imidazol-2-yl )cyclopentyl)carbamoyl)-l- (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy) butanoic acid:

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy- 5a,5b,8,8, 11 a-pentamethyl-N-( 1 -(5-phenyl- 1 H-imidazol-2-yl)cyclopentyl)- 1 -(prop- 1 -en-2- yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide (step 2, 0.06 g, 0.09 mmol) and 2,2-dimethyl succinicanhydride (0.046 g, 0.36 mmol) in toluene (4 ml), was added DMAP (0.022 g, 0.18 mmol). The reaction mixture was heated at 90°C for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure, cooled to 0°C, acidified to pH=6 with IN HC1 and extracted with DCM. The combined organic extracts were washed with water, brine, dried over Na ₂ S0 ₄ then the solvent was evaporated under reduced pressure and the resulting solid was recrystallized from ACN to afford the title compound (0.022 g, yield: 31%). 1H NMR (300 MHz, CDC1 ₃ ): δ 7.65-7.62 (m, 2H), 7.36-7.29 (m, 4H), 4.59 (s, 1H), 4.51 (s, 1H), 4.46-4.38 (m, 1H), 2.93- 2.91 (m, 1H), 2.62-1.31 (m, 32H), 1.26 (s, 3H), 1.22 (s, 6H), 0.86-0.68 (m, 19H); ES Mass: 794.64 [M+H] ⁺ .

Example- 19: Preparation of. ( lR,3S)-2,2-dimethyl-3-

((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8,l la-pentamethyl-3a-((l-(5- phenyl-lH-imidazol-2-yl)cvclopropyl)carbamoyl)-l-(prop-l-en- 2-yl)icosahvdro-lH- cvclopentaralchrvsen-9-yl)oxy)carbonyl)cvclobutane-l-carboxy lic acid

Step 1: Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla- pentamethyl-3a-((l -(5-phenyl-lH-imidazol-2-yl)cyclopropyl)carbamoyl)-l -(prop-1 -en-2- yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate

To a stirred solution of l-(5-phenyl-lH-imidazol-2-yl)cyclopropan-l -amine hydrochloride (Intermediate 7, 0.534 g, 2.3 mmol) in DCM (6 ml) and NEt ₃ (2.0 ml, 13.5 mmol), was added a solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a- (chlorocarbonyl)-5a,5b,8,8, 1 la-pentamethyl- l-(prop- l-en-2-yl)icosahydro- 1H- cyclopenta[a]chrysen-9-yl acetate (prepared as described in WO 2013/160810 A2, 1.0 g, 1.9 mmol) in DCM (12 ml) at 0 C and allowed to stir at room temperature for about 16 h. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, brine and dried over Na ₂ S0 ₄ , and the solvent was evaporated and purified by silica gel column (elution 30 % EtOAc in n- hexane) to afford the title compound as an off white solid (0.8 g, Yield: 58.8%). 1H NMR (300 MHz, DMSO-d ₆ ): δ 11.28 (s, 1H), 8.38 (s, 1H), 7.73-7.70 (m, 2H), 7.40 (s, 1H), 7.31-7.26 (m, 2H), 7.15-7.10 (m, 1H), 4.62 (s, 1H), 4.51 (s, 1H), 4.37-4.32 (m, 1H), 3.00-2.94 (m, 1H), 2.27-2.24 (m, 1H), 2.05-1.95 (m, 1H), 1.99 (s, 3H), 1.80-1.74 (m, 1H), 1.62-1.00 (m, 24H), 0.91-0.72 (m, 19H). ESI Mass: 680.55 [M+H] ⁺

Step 2: Synthesis of (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-9-hydroxy-

5a,5b,8,8,lla-pentamethyl-N-(l-(5-phenyl-lH-imidazol-2-yl )cyclopropyl)-l-(prop-l-en-2- yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-

5a,5b, 8 ,8 , 11 a-pentamethyl-3 a-(( 1 -(5-phenyl- 1 H-imidazol-2-yl)cyclopropyl)carbamoyl) - 1 - (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (0.8 g, 1.2 mmol) in MeOH: THF (5 ml: 5 ml), was added 4N NaOH (8 ml) at 0 °C and allowed to stir at room temperature for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure. The reaction mixture was diluted with DCM and washed with water, brine and dried over Na ₂ S0 ₄ . The solvent was evaporated and the resulting solid was taken in hexane and stirred for one hour and filtered to afford the title compound as a white solid (0.65 g, yield: 86.6%).

Step 3: Synthesis of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)- 5a,5b,8,8,lla-pentamethyl-3a-((l-(5-phenyl-lH-imidazol-2-yl) cyclopropyl)carbamoyl)-l- (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2- dimethylcyclobutane-l -dicarboxylate

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy- 5a,5b,8,8, 11 a-pentamethyl-N-( 1 -(5-phenyl- 1 H-imidazol-2-yl)cyclopropyl)- 1 -(prop- 1 -en-2- yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide (0.65 g, 1.0 mmol) and (lS,3R)-3- ((benzyloxy)carbonyl)-2,2-dimethylcyclobutane-l-carboxylic acid (0.4 g, 1.5 mmol) and DMAP (0.02 g, cat) in DCM (10 ml), was slowly added DCC (0.42 g, 2.0 mmol) in DCM (5 ml) at 0°C and allowed to stir at room temperature for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHC0 ₃ solution, brine and dried over Na ₂ S0 ₄ . The solvent was evaporated and to the resulting solid, was added DCM (4 ml) and stirred for one hour and filtered. The filtrate was concentrated under reduced pressure to afford the title compound as a white solid (0.6 g, yield: 53.4%).

Step 4: Synthesis of (lR,3S)-2,2-dimethyl-3-

((((1R, 3aS, 5aR, 5bR, 7aR, 9S, llaR l bR, 13aR, 13bR ) -5 a, 5b, 8, 8, 1 la-pentamethyl-3a-((l -(5- phenyl-lH-imidazol-2-yl)cyclopropyl)carbamoyl)-l-(prop-l-en- 2-yl)icosahydro-lH- cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxy lic acid

To a stirred solution of 1-benzyl 3- ((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8,l la-pentamethyl-3a-((l-(5- phenyl-lH-imidazol-2-yl)cyclopropyl)carbamoyl)-l-(prop-l-en- 2-yl)icosahydro-lH- cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-dimethylcyclobutane-l,3-dicarboxylate (0.6 g, 0.68 mmol) in THF (10 ml), were added N(Et) ₃ (0.4 ml, 2.9 mmol), triethyl silane (0.4 ml, 2.6 mmol) and Pd(OAc) ₂ (0.03 g, cat). The reaction mixture was heated to 70 °C and continued for 12 h. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc and washed with water, saturated NaHC0 ₃ solution, brine and dried over Na ₂ S0 ₄ . The solvent was evaporated under reduced pressure and purified by silica gel column (elution 5 % MeOH in DCM) to afford the title compound as a white solid (0.32 g, Yield: 59.5%). 1H NMR (300 MHz, DMSO-d ₆ ): δ 12.13 (s, 1H), 11.28 (s, 1H), 8.38 (s, 1H), 7.73-7.70 (m, 2H), 7.40 (s, 1H), 7.31-7.26 (m, 2H), 7.15-7.10 (m, 1H), 4.62 (s, 1H), 4.51 (s, 1H), 4.34-4.29 (m, 1H), 3.02-2.96 (m, 1H), 2.81-2.72 (m, 2H), 2.30-2.23 (m, 2H), 1.99-1.11 (m, 33H), 0.99-0.74 (m, 19H). ESI Mass: 792.58 [M+H] ⁺ .

Example 20j Preparation of of 2,2-dimethyl-4-oxo-4- (((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8,l la-pentamethyl-3a-((l-(5- phenyl-lH-imidazol-2-yl)cvclopropyl)carbamoyl)-l-(prop-l-en- 2-yl)icosahvdro-lH- cvclopentaralchrvse -9-yl)oxy)butanoic acid

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy- 5a,5b,8,8, 11 a-pentamethyl-N-( 1 -(5-phenyl- 1 H-imidazol-2-yl)cyclopropyl)- 1 -(prop- 1 -en-2- yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide (Step 2 Example 19, 0.2 g, 0.31 mmol) and 2,2-dimethyl succinicanhydride (0.16 g, 1.2 mmol) in toluene (7 ml), was added DMAP (0.076 g, 0.6 mmol). The reaction mixture was heated at 90 °C for 12 h. After completion of the reaction (monitored by TLC), reaction mixture was concentrated under reduced pressure, cooled to 0 °C, acidified to pH = 6 with IN HC1 and extracted with DCM. The combined organic extracts was washed with water, brine, dried over Na ₂ S0 ₄ then the solvent was evaporated under reduced pressure and to the resulting solid was recrystallized from ACN to give the title compound as a white solid (0.11 g, Yield: 45.83%). 1H NMR (300 MHz, DMSO-d ₆ ): δ 12.15 (s, 1H), 11.27 (s, 1H), 8.37 (s, 1H), 7.72-7.70 (m, 2H), 7.39 (s, 1H), 7.31-7.26 (m, 2H), 7.15-7.11 (m, 1H), 4.61 (s, 1H), 4.51 (s, 1H), 4.37-4.32 (m, 1H), 3.00-2.94 (m, 1H), 2.27 (m, 1H), 1.98-1.95 (m, 1H), 1.79-1.73 (m, 1H), 1.62-1.00 (m, 32H), 0.91-0.74 (m, 19H). ESI Mass: 766.56 [M+H] ⁺ .

Example -21 : Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((l-(l-(2-(dimethylamino)ethyl)-4-phenyl-lH-imidazol-2-yl )cvclopropyl)carbamoyl)- 5a,5b,8,8 J la-pentamethyl-l-(prop-l-en-2-yl)icosahvdro-lH-cvclopentaral chrvsen-9- yl)oxy)carbonyl)-2,2-dimethylcyclobutane- 1-carboxylic acid

Step 1: Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-3a-((l-(l-(2- (dimethylamino)ethyl)-4-phenyl-lH-imidazol-2-yl)cyclopropyl) carbamoyl)-5a,5b,8,8,lla- pentamethyl-1 -(prop-1 -en-2-yl)icosahydro-lH-cyclopenta[a] chrysen-9-yl acetate

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 5a,5b, 8 ,8 , 11 a-pentamethyl-3 a-(( 1 -(5-phenyl- 1 H-imidazol-2-yl)cyclopropyl)carbamoyl) - 1 - (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (Step 1 Example 19, 2.0 g, 2.9 mmol) in DMF (20 ml) at 0°C, were added Cs ₂ C0 ₃ (2.87 g, 8.8 mmol) and 2-chloro-N,N- dimethylethan-1- amine hydrochloride (0.63 g, 4.4 mmol). The reaction mixture was stirred for overnight at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc and washed with water, brine and dried over Na ₂ S0 ₄ . The solvent was evaporated under reduced pressure and purified by silica gel column (elution 3% MeOH in DCM) to afford the title compound as a white solid (1.0 g, Yield: 45%).

Step 2: Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-N-(l-(l-(2- ( dimethylamino )ethyl ) -4-phenyl-lH-imidazol-2 -yl)cyclopropyl)-9-hydroxy-5a, 5b, 8,8,11a- pentamethyl-1 -(prop-1 -en-2-yl)icosahydro-3aH-cyclopenta[a] chrysene-3a-carboxamide

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(l- (2-(dimethylamino)ethyl) -4-phenyl- 1 H-imidazol-2-yl)cyclopropyl)carbamoyl)-5 a,5b, 8 ,8 , 11a- pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chr ysen-9-yl acetate (0.64 g, 0.85 mmol) in MeOH: THF (5 ml: 5 ml), was added 8N NaOH (5 ml) at 0 °C and allowed to stir at room temperature for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure. The reaction mixture was diluted with DCM and washed with water, brine and dried over Na ₂ S0 ₄ . The solvent was evaporated and the resulting solid was taken in hexane and stirred for one hour and filtered to afford the title compound as a white solid (0.5 g, yield: 83%). 1H MR (300 MHz, DMSO- d ₆ ): δ 8.34 (s, 1H), 7.70-7.68 (m, 2H), 7.54 (s, 1H), 7.34-7.29 (m, 2H), 7.17-7.12 (m, 1H), 4.61 (s, 1H), 4.51 (s, 1H), 4.37-4.13 (m, 3H), 3.28 (m, 1H), 2.95-2.93 (m, 1H), 2.63-2.61 (m, 2H), 2.22 (s, 6H), 2.13-2.11 (m, 1H), 1.76-1.03 (m, 26H), 0.94-0.63 (m, 19H).

Step 3: Synthesis of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-3a-((l-(l- (2-(dimethylamino)ethyl)-4^henyl-lH-imidazol-2-yl)cyclopropy l)carbamoyl)-5a,5b,8,8,lla- pentamethyl-1 -(prop-1 -en-2-yl)icosahydro-lH-cyclopenta[a] chrysen-9-yl) (lR,3S)-2,2- dimethylcyclobutane-l,3-dicarboxylate

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-N-(l-(l-(2- (dimethylamino)ethyl)-4-phenyl-lH-imidazol-2-yl)cyclopropyl) -9-hydroxy-5a,5b,8,8,l la- pentamethyl-l-(prop-l-en-2-yl)icosahydro-3aH-cyclopenta[a]ch rysene-3a-carboxamide (0.5 g, 0.71 mmol) and (lS,3R)-3-((benzyloxy)carbonyl)-2,2-dimethylcyclobutane-l-ca rboxylic acid (prepared as described in WO 2013/160810 A20.28 g, 1.0 mmol) and DMAP (0.02 g, cat) in DCM (8 ml), was slowly added DCC (0.29 g, 1.4 mmol) in DCM (4 ml) at 0°C and allowed to stir at room temperature for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHC0 ₃ solution, brine and dried over Na ₂ S0 ₄ . The solvent was evaporated and to the resulting solid, was added DCM (4 ml) and stirred for one hour and filtered. The filtrate was concentrated under reduced pressure to afford the title compound as a white solid (0.4 g, yield: 60%). ESI Mass: 953.94 [M+H] ⁺ .

Step 4: Synthesis of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,llaR,nbR,13aR,13bR)-3a-( (l-(l- (2-(dimethylamino)ethyl)-4^henyl-lH-imidazol-2-yl)cyclopropy l)carbamoyl)-5a,5b,8,8,lla- pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chr ysen-9-yl)oxy)carbonyl)-2,2- dimethylcyclobutane-l -carboxylic acid

To a stirred solution of 1 -benzyl 3-

((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(l-(2-(dimethylamino)ethyl)-4- phenyl-lH-imidazol-2-yl)cyclopropyl)carbamoyl)-5a,5b, 8,8,1 la-pentamethyl-l-(prop-l-en-

2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) ( 1 R,3 S )-2,2-dimethylcyclobutane- 1,3- dicarboxylate (0.4 g, 0.42 mmol) in THF (8 ml), were added N(Et) ₃ (0.3 ml, 1.6 mmol), triethyl silane (0.3 ml, 1.3 mmol) and Pd(OAc) ₂ (0.03 g, cat). The reaction mixture was heated to 70 °C and continued for 12 h. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc and washed with water, saturated NaHC0 ₃ solution, brine and dried over Na ₂ S0 ₄ . The solvent was evaporated under reduced pressure and purified by silica gel column (elution 5 % MeOH in DCM) to afford the title compound as a white solid (0.16 g, Yield: 44.4%). 1H MR (300 MHz, DMSO-d ₆ ): δ 12.13 (brs, 1H), 8.33 (s, 1H), 7.70-7.68 (m, 2H), 7.54 (s, 1H), 7.34-7.29 (m, 2H), 7.17-7.12 (m, 1H), 4.62 (s, 1H), 4.51 (s, 1H), 4.32-4.29 (m, 2H), 4.24-4.17 (m, 1H), 2.95-2.90 (m, 1H), 2.81-2.72 (m, 2H), 2.64-2.58 (m, 2H), 2.30-2.14 (m, 2H), 2.23 (s, 6H), 1.91-1.04 (m, 31H), 0.98-0.73 (m, 21H). ESI Mass: 863.63 [M+H] ⁺ .

Example 22: Preparation of _. ( lR,3S)-2,2-dimethyl-3-

((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8,l la-pentamethyl-3a-(((5- phenyl-lH-imidazol-2-yl)methyl)carbamoyl)-l-(prop-l-en-2-yl) icosahvdro-lH- cyclopentaralchrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxy lic acid

Step 1: Synthesis of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)- 5a,5b,8,8 la^entamethyl-3a-(((5-phenyl-lH-imidazol-2-yl)methyl)carbamo yl)-l-(prop-l- en-2-yl)icosahydro-l H-cyclopenta[a] chrysen-9-yl) (lR,3S)-2,2-dimethylcyclobutane-l,3- dicarboxylate

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy- 5a,5b,8,8,l la-pentamethyl-N-((5-phenyl-lH-imidazol-2-yl)methyl)-l-(prop -l-en-2- yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide (1.0 g, 1.64 mmol, 1.0 eq) in toluene (10 ml) was added DMAP (0.39 g, 3.27 mmol, 2.0 eq) and (IS, 3R)-3- (benzyloxycarbonyl)-2,2-dimethylcyclobutane carboxylic 2,4,6-trichlorobenzoic anhydride (1.54 g, 3.27 mmol, 2.0 eq). The reaction mixture was heated to 90 °C for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was evaporated under reduced pressure and diluted with water. The aqueous layer was extracted with CH ₂ CI ₂ (2x100 ml). The combined organic layers were dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 70% EtOAc: n-Hexane as an eluent to obtain (1.08 g, 76.9 % yield) as brown color solid. 1H NMR (300 MHz, DMSO): δ 8.11 (bs, 1H), 7.61 (d, J = 7.2 Hz, 2H), 7.39 (t, J = 7.2 Hz, 2H), 7.31-7.22 (m, 7H), 5.08 (ABq, J = 3.9, 12.3 Hz, 2H), 4.7 (s, 1H), 4.68 (s, 1H), 4.41 -4.36 (m, 2H), 3.06-3.01 (m, 1H), 2.81-2.57 (m, 3H), 2.4-2.12 (m, 2H), 2.1-1.72 (m, 3H), 1.68-1.31 (m, 13H), 1.3-1.01 (m, 12H), 1.0-0.98 (m, 1H), 0.96-0.83 (m, 8H), 0.81-0.76 (m, 6H), 0.73-0.61 (m, 7H). ES Mass: [M+l] ⁺ 856.

Step 2 : (1R, 3S)-2, 2-dimethyl-3-((((lR, 3aS, 5aR, 5bR, 7aR, 9S, llaR l bR, 13aR, 13bR)- 5a,5b,8,8 la^entamethyl-3a-(((5-phenyl-lH-imidazol-2-yl)methyl)carbamo yl)-l-(prop-l- en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl )cyclobutane-l -carboxylic acid

To a stirred solution of 1 -benzyl 3-

((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8,l la-pentamethyl-3a-(((5- phenyl-lH-imidazol-2-yl)methyl)carbamoyl)-l-(prop-l-en-2-yl) icosahydro-lH- cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-dimethylcyclobutane-l,3-dicarboxylate (0.82 g, 0.96 mmol, 1.0 eq) in EtOAc: MeOH (1 : 1, 16 mL) added ammonium formate (302 mg, 4.78 mmol, 5.0 eq), 10% Pd/C (100 mg) and stirred the reaction mixture for 2 hr. The resultant mixture was filtered through celite and washed with methanol, collected filtrate was concentrated and obtained crude residue was dissolved in EtOAc (100 mL) and washed with water, brine and organic layer was concentrated under reduced pressure to get the residue, which was purified by silicagel column chromatography by using 70% EtOAc: n-Hexane as an eluent gave the desired compound (0.51 g, 69.4 % yield) as a off- white solid. 1H MR (300 MHz, DMSO): δ 12.15 (bs, 1H), 11.66 (bs, 1H), 8.1 (bs, 1H), 7.73 (d, J = 6.9 Hz, 2H), 7.45 (s, 1H), 7.3 (t, J = 6.9 Hz, 2H), 7.16-7.14 (m, 1H), 4.64 (s, 1H), 4.53 (s, 1H), 4.34-4.21 (m, 3H), 3.41-3.37 (m, 1H), 3.07-2.98 (m, 1H), 2.81-2.72 (m, 2H), 2.38-2.21 (m, 2H), 1.91- 1.7 (m, 3H), 1.69-1.3 (m, 11H), 1.29-1.13 (m, 9H), 1.11-0.98 (m, 4H), 0.92-0.86 (m, 8H), 0.8-0.61 (m, 12H). ES Mass: [M+l] ⁺ 766.5. HPLC purity: 96.52%.

Example 23j Preparation of 2,2-dimethyl-4-oxo-4-

(((lR.3aS.5aR.5bRJaR.9S.l laR.l lbR.13aR.13bR)-5a.5b.8.8.11a-pentamethyl-3a-(((5- phenyl- lH-imidazol-2-yl)methyl)carbamoyl)-l -(prop- 1 -en-2-yl)icosahydro- 1H- cyclopenta[alchrysen-9-yl)oxy)butanoic acid

To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy- 5a,5b,8,8,l la-pentamethyl-N-((5-phenyl-lH-imidazol-2-yl)methyl)-l-(prop -l-en-2- yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide (0.6 g, 0.98 mmol, 1.0 eq) and 2,2-dimethyl succinic anhydride (0.5 g, 3.92 mmol, 4.0 eq) in toluene (12 mL) was added

DMAP (0.239 g, 1.96 mmol, 2.0 eq). The reaction mixture was heated at 90 °C for overnight. TLC indicated starting material was consumed and the desired product was observed. The mixture was concentrated under reduced pressure, cooled to 0 °C, acidified to pH= 6-7 with IN HC1 and extracted with CH ₂ CI ₂ . The combined organic extracts were washed with water, dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The crude residue was purified by silicagel column chromatography by using 60% EtOAc: n-Hexane as an eluent gave the desired product (0.13 g, 18% yield) as brown color solid. 1H MR (300 MHz, DMSO): δ 12.2 (bs, 1H), 11.66 (bs, 1H), 8.13-8.11 (m, 1H), 7.73 (d, J = 6.3 Hz, 2H), 7.32 (s, 1H), 7.3 (t, J = 6.9Hz, 2H), 7.17-7.14 (m, 1H), 4.64 (s, 1H), 4.52 (s, 1H), 4.37-4.22 (m, 4H), 3.05-2.98 (m, 1H), 2.31-2.18 (m, 2H), 1.91-1.61 (m, 6H), 1.6-1.21 (m, 18H), 1.09-0.96 (m, 8H), 0.92 (s, 3H), 0.81-0.63 (m, 12H). ES Mass: [M+l] ⁺ 740.55. HPLC: 86.0%.

PHARMACOLOGICAL ACTIVITY

The compounds described herein can be tested for their biological activity following procedures known to a person of ordinary skill in the art. For example, the following protocols can be employed for testing the compounds. These protocols are illustrative and do not limit to the scope of the invention.

Example 24: Evaluation of compounds antiviral activity:

MT2 cells were infected with HIV-1 strain 92HT599 (10 TCID 50/ 30000 cells). The infected cells were plated at the concentration of -30,000 cells per well in 96 well plate. Test compound was added to the micro plate in defined format with the final concentration of DMSO (vehicle) is not more than 1%. Incubation was carried out in C0 ₂ incubator for ~ 96 hours for viral infection. At the end of incubation period an aliquot from each well was taken for p24 estimation. The quantitation of p24 is an index for antiviral activity of the compound. Percent inhibition was calculated with reference to control values (vehicle controls).

P-24 estimation was carried out using Advance biosciences kit as per the procedure detailed by supplier.

For 0% serum binding assay, wherein "A" refers to an IC ₅₀ value < 10 nM, "B" refers to IC ₅₀ value in range of 10.01-50 nM, and "C" refers to IC ₅₀ value > 50 nM.

For 45 mg/ml HSA serum binding assay, wherein "A" refers to an IC ₅₀ value < 50 nM, "B" refers to IC ₅₀ value in range of 50.01-200 nM, and "C" refers to IC ₅₀ values > 100 nM. The IC ₅₀ (nM) values are set forth in Table- 1. TABLE- 1

References:

1. Antiviral methods and protocols (Eds: D Kinchington and R F Schinazi) Humana Press Inc., 2000.

2. HIV protocols (Eds: N L Michael and J H Kim) Humana Press Inc, 1999.

3. DAIDS Virology manual from HIV laboratories, Publication NIH-97-3838, 1997.

4. HIV-1 p24 antigen capture assay, enzyme immunoassay for detection of Human immunodeficiency Virus Type 1 (HIV-1) p24 in tissue culture media - Advanced bio science laboratories, Inc kit procedure.

Although the invention herein has been described with reference to particular embodiments, it is to be understood that these embodiments are merely illustrative of the principles and applications of the present invention. It is therefore to be understood that numerous modifications may be made to the illustrative embodiments and that other arrangements may be devised without departing from the spirit and scope of the present invention as described above.

All publications and patent applications cited in this application are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated herein by reference.