WO2017025899A1 - Novel c28-amides with c3-modifications of triterpene derivatives as hiv inhibitors - Google Patents

Abstract

Formula (I) The present invention relates to novel C28-amides with C3-modifications of triterpene compounds of formula (I); or pharmaceutically acceptable salts, pharmaceutically acceptable solvates, pharmaceutically acceptable hydrates, tautomers, stereoisomers, prodrugs or combination thereof, wherein R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, R₉, X, and Y are as defined herein. The present invention also relates to pharmaceutical compositions comprising compounds of formula (I) useful for the therapeutic treatment of viral mediated diseases and particularly HIV mediated diseases.

Description

NOVEL C28- AMIDES WITH C3-MODIFICATIONS OF TRITERPENE DERIVATIVES AS HIV INHIBITORS

This application claims the benefit of Indian provisional application no 4181/CHE/2015 filed on 11^th August 2015 which is hereby incorporated by reference in its entirety.

FIELD OF THE INVENTION

The present invention relates to novel C28-amides with C3 -modifications 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-re spondant 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, researchers started exploring betulinic acid derivatives and related compounds intensively for their therapeutic activities. For example, WO 2016/001820 disclosed novel betulinic proline imidazole derivatives as HIV inhibitors; WO 2015/198263 disclosed novel betulinic proline substituted derivatives as hiv inhibitors; WO 2014/105926 disclosed novel betulinic acid proline derivatives as HIV inhibitors; WO 2014/130810 disclosed C-3 alkyl and alkenyl modified betulinic acid derivatives useful in the treatment of HIV; WO 2014/123889 disclosed C-19 modified triterpenoids with HIV maturation inhibitory activity; WO 2013/160810 disclosed novel betulinic acid derivatives as HIV inhibitors; WO 2013/169578 disclosed C-17 bicyclic amines of triterpenoids with HIV maturation inhibitory activity; WO 2013/123019 disclosed C-3 cycloalkenyl triterpenoids with HIV maturation inhibitory activity; WO 2012/106190 disclosed C-17 and C-3 modified triterpenoids with HIV maturation inhibitory activity; WO 2012/106188 disclosed C-28 amines of C-3 modified betulinic acid derivatives as HIV maturation inhibitors; WO 2011/007230 disclosed lupeol-type triterpene derivatives as antivirals; WO 2011/153319 disclosed modified C-28 amides of modified C-3 betulinic acid derivatives as HIV maturation inhibitors; WO 2011/153315 disclosed modified C-3 betulinic acid derivatives as HIV maturation inhibitors; WO 2009/100532 disclosed novel 17 β lupane derivatives as anti-HIV agents. The patent publication WO 2008/057420 describes extended triterpene derivatives as antiretroviral agents; WO 2008/127364 describes preparation of antiviral compounds and use thereof; WO 2007/141392 describes compositions comprising betulonic acid; WO 2007/141391 describes betulin derived compounds useful as antiprotozoal agents; WO 2007/141390 describes preparation of betulin derived compounds as antiviral agents; WO 2007141389 describes betulin derived compounds as antibacterial agents; WO 2007/002411 describes antiviral compounds; WO 2006/053255 describes novel betulin derivatives, preparation and use thereof and WO 2006/105356 describes methods of manufacturing bioactive 3-esters of betulinic aldehyde and betulinic acid; US 2004/0204389 describes anti- HIV agents with dual sites of action; Pharmazie (1998), 53(10), 677-680 describes synthesis of amino acid conjugates of 3a-hydroxylup-20(29)-ene-23,28-dioic acid and Journal of Medicinal Chemistry (1996), 39(5), 1056-68 describes betulinic acid derivatives: a new class of human immunodeficiency virus type 1 specific inhibitors with a new mode of action.

Some additional references disclose betulinic acid related compounds. For example, WO 2007/141383 describes betulin derivatives as antifeedants for plant pests; US 6670345 describes use of betulinic acid and its derivatives for inhibiting cancer growth and process for the manufacture of betulinic acid; WO 2002/091858 describes anxiolytic marcgraviaceae compositions containing betulinic acid, betulinic acid derivatives, and methods of preparation and use; WO 2000/046235 describes preparation of novel betulinic acid derivatives for use as cancer growth inhibitors; and Pharmaceutical Chemistry Journal, 2002, 36(9), 29-32 describes synthesis and anti-inflammatory activity of new acylated betulin 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 compounds of the formula (I):

Formula (j) wherein,

Ri can be substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkadienyl,

substituted or unsubstituted aryl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted heteroaryl (preferably, pyridine, pyrazine, pyrimidine, oxazole or thiazole), or substituted or unsubstituted fused bicycle; wherein R_b at each occurrence can be hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl;

R₂ can be hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted aminoacid, substituted or unsubstituted alkoxy, or substituted or unsubstituted cycloalkyl;

X can be absent, O, S, CH₂ or NR_a; (wherein R_a can be hydrogen, 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 can be taken together to form N-contained heterocyclyl (Preferably, pyrrolidine, piperdine, piperzine, or morpholine));

Y can be absent, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heteroaryl, or -(CR_eRf)_m;

R₃ and R₄ can be independently selected from hydrogen, hydroxy, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted alkoxylalkoxy, or substituted or unsubstituted aminoacid; wherein amino acid is substituted by substituted or unsubstituted alkyl; or R₃ and R₄ can be taken together with their carbon atoms to which they are attached to form a bond; or R₃ and R₄ can be taken together with their carbon atoms to which they are attached to form cycloalkyl; or R₃ and R₄ can be taken together with the carbon atoms to which they are attached to form epoxide;

R5 and R_c can be independently selected from hydrogen, C(0)₂R_d (wherein R_d can be hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl), or substituted or unsubstituted alkyl;

R₆, R7 and R₈ can be independently selected from hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl; or R₆ and R₇ can be taken together with the nitrogen and carbon to which they are attached to form substituted or unsubstituted 4-10 membered heterocyclyl, or substituted or unsubstituted 5-15 membered bridged heterocyclyl; or R₇ and R₈ can be taken together with the carbon atom to which they are attached to form 3-7 membered cycloalkyl or heterocyclyl;

R9 can be absent, substituted or unsubstituted amine, substituted or unsubstituted 4-10 membered heterocyclyl, substituted or unsubstituted 4-10 membered heteroaryl,- C(0)heterocyclyl or substituted or unsubstituted aryl; wherein the substituent can be alkyl;

R_e and R_f can be independently selected from hydrogen, or substituted or unsubstituted alkyl;

'n' can be an integer from 0-1; and 'm' can be an integer from 0-2;

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, the present invention relates to compound of formula (I), wherein Ri is benzoic acid.

According to other embodiment, the present invention relates to compound of formula (I), wherein 'X' is absent.

According to yet another embodiment, the present invention relates to compound of formula (I), wherein R₂ is hydrogen.

According to yet another embodiment, the present invention relates to compound of formula (I), when "X is absent, and R₂ and R5 are hydrogen"; then R₃ and R₄ are taken together with carbon atoms to which they are attached to form a bond.

According to yet another embodiment, the present invention relates to compound of formula (I), when "X is absent, and R₂ and R5 are hydrogen"; then R₃ and R₄ are taken together with carbon atoms to which they are attached form cycloalkyl (preferably cyclopropyl).

According to yet another embodiment, the present invention relates to compound of formula (I), wherein R₆ is hydrogen or alkyl (preferably methyl).

According to yet another embodiment, the present invention relates to compound of formula (I), wherein R₆ and R₇ are taken together with the nitrogen and carbon atoms to which they are attached to form a substituted or unsubstituted 4-10 membered heterocyclyl.

According to yet another embodiment, the present invention relates to compound of formula (I), wherein the above said heterocyclyl is substituted or unsubstituted pyrrolidine.

According to yet another embodiment, the present invention relates to compound of formula (I), wherein R₆ and R₇ are taken together with the nitrogen and carbon atoms to which they are attached to form a substituted or unsubstituted 5-15 membered bridged heterocyclyl. According to yet another embodiment, the present invention relates to compound of formula (I), wherein the above said 5-15 membered bridged heterocyclyl is (S)- hexahydropyrrolo [ 1 ,2- a] pyrazin-4( 1 H) -one .

According to yet another embodiment, the present invention relates to compound of formula (I), wherein the above said 5-15 membered bridged heterocyclyl is (S)-octahydro- 4H-pyrido[ 1 ,2-a]pyrazin-4-one.

According to yet another embodiment, the present invention relates to compound of formula (I), wherein Y is absent.

According to yet another embodiment, the present invention relates to compound of formula (I), wherein Ύ' is CR_eR_f; wherein R_e and R_f are hydrogen.

According to yet another embodiment, the present invention relates to compound of formula (I), wherein Ύ' is substituted or unsubstituted cycloalkyl.

According to yet another embodiment, the present invention relates to compound of formula (I), wherein the above said substituted or unsubstituted cycloalkyl is cyclobutyl substituted with one or more alkyl (preferably, methyl).

According to yet another embodiment, the present invention relates to compound of formula (I), wherein Ύ' is substituted or unsubstituted heteroaryl (preferably, imidazole).

According to yet another embodiment, the present invention relates to compound of formula (I), wherein R9 is absent.

According to yet another embodiment, the present invention relates to compound of formula (I), wherein R9 is substituted or unsubstituted 4-10 membered heterocyclyl.

According to yet another embodiment, the present invention relates to compound of formula (I), wherein the above said 4-10 membered heterocyclyl is pyrrolidine.

According to yet another embodiment, the present invention relates to compound of formula (I), wherein the above said 4-10 membered heterocyclyl is morpholine.

According to yet another embodiment, the present invention relates to compound of formula (I), wherein the above said 4-10 membered heterocyclyl is piperidine.

According to yet another embodiment, the present invention relates to compound of formula (I), wherein the above said 4-10 membered heterocyclyl is piperazine substituted with alkyl (preferably, ethyl).

According to yet another embodiment, the present invention relates to compound of formula (I), wherein R9 is diethyl amine.

According to yet another embodiment, the present invention relates to compound of formula (I), wherein R9 is substituted or unsubstituted 4-10 membered heteroaryl. According to yet another embodiment, the present invention relates to compound of formula (I), wherein the above said 4-10 membered heteroaryl is 1,3,4-triazole substituted with alkyl (preferably, methyl and isopropyl).

According to yet another embodiment, the present invention relates to compound of formula (I), wherein R9 is substituted or unsubstituted aryl (preferably, phenyl).

According to yet another embodiment, the present invention relates to compound of formula (I), wherein R9 is -C(0)heterocyclyl (preferably,

According to yet another embodiment, the present invention relates to compound of formula (I), wherein 'n' is an integer from 0-1.

According to yet another embodiment, the present invention relates to compound of formula (I), wherein 'm' is an integer from 0-2.

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 compounds, 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):

4-((lR,3aS,5aR,5bR,7aR,l laS,l lbR,13aR,13bR)-5a,5b,8,8,l la-pentamethyl-l-(l- methylcyclopropyl)-3a-((S)-2-(5-phenyl-lH-imidazol-2-yl)pyrrolidine-l-carbonyl)-2,3,3a, 4,5,5a,5b,6,7,7a,8,l 1,1 la,l lb, 12,13, 13a,13b-octadecahydro-lH-cyclopenta[a]chrysen-9- yl)benzoic acid (Example 1);

4-((lR,3aS,5aR,5bR,7aR,l laS,l lbR,13aR,13bR)-3a-(((lR,3S)-2,2-dimethyl-3- (piperidine-l-carbonyl)cyclobutyl)carbamoyl)-5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclo propyl)-2,3,3a,4,5,5a,5b,6,7,7a,8,l 1,1 la,l lb, 12,13, 13a,13b-octadecahydro-lH-cyclopenta[a] chrysen-9-yl)benzoic acid (Example 2);

4-((lR,3aS,5aR,5bR,7aR,l laS,l lbR,13aR,13bR)-3a-(dimethylcarbamoyl)-5a,5b,8, 8,1 la-pentamethyl-l-(l-methylcyclopropyl)-2,3,3a,4,5,5a,5b,6,7,7a,8,l 1,1 la,l lb, 12,13, 13a, 13b-octadecahydro-lH-cyclopenta[a]chrysen-9-yl)benzoic acid (Example 3); and

4-(lR,3aS,5aR,5bR,7aR,l laS,l lbR,13aR,13bR)-5a,5b,8,8,l la-pentamethyl-l-(l- methylcyclopropyl)-3a-((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidine-l-carbonyl)-2,3,3a,4,5,5a, 5b,6,7,7a,8,l 1,1 la,l lb, 12,13, 13a,13b-octadecahydro-lH-cyclopenta[a]chrysen-9-yl)benzoic acid (Example 4), 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 an 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 novel C28-amides with C3-modifications 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 "alkenyl" refers to a hydrocarbon chain that may be a straight chain or branched chain, containing the indicated number of carbon atoms having at least one -C=C-, for example, a C₂-C6 alkenyl group may have from 2 to 6 (inclusive) -C=C- atoms in it. Examples of C₂-C₆ alkenyl groups include, but are not limited to ethylene, prop-l-ene, but-1- ene, but-2-ene, pent-l-ene, pent-2-ene, hex-l-ene, hex-2-ene and the like.

The term "alkadienyl" refers to a hydrocarbon chain that may be a straight chain and branched, preferably straight chain, saturated, mono-unsaturated and di-unsaturated, respectively, hydrocarbon radicals. Examples of alkadienyl groups are butadienyl, pentadienyl, hexadienyl, heptadienyl, heptadecadienyl.

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 "amino" or "amine" refers to -NH₂.

The term "alkoxylalkoxy" refers to a straight or branched hydrocarbon chain with oxygen radical consisting carbon atom, hydrogen atom and alkoxy groups, 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., 2- (methyloxy)ethyloxy, 2-(ethyloxy)ethyloxy, 2-(n-propyloxy)ethyloxy, and 3- (isopropyloxy)butyloxy.

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 "cycloalkyl" denotes a non-aromatic mono or multicyclic ring system of from 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 "aryl" refers to an aromatic radical having from 6 to 14 carbon atoms such as phenyl, naphthyl, tetrahydronapthyl, indanyl, and biphenyl.

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; and the ring radical may be partially or fully saturated (i.e., heterocyclic or heteroaryl). Examples of such heterocyclic ring radicals include, but are not limited to, tetrazoyl, tetrahydroisouinolyl, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxoazepinyl, azepinyl, pyrrolyl, 4-piperidonyl, pyrrolidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolinyl, oxazolidinyl, triazolyl, isoxazolyl, isoxasolidinyl, morpholinyl, thiazolyl, thiazolinyl, thiazolidinyl, isothiazolyl, quinuclidinyl, isothiazolidinyl, indolyl, isoindolyl, indolinyl, isoindolinyl, octahydroindolyl, octahydroisoindolyl, quinolyl, isoquinolyl, decahydroisoquinolyl, benzimidazolyl, thiadiazolyl, benzothiazolyl, benzooxazolyl, furyl, tetrahydrofurtyl, tetrahydropyranyl, thienyl, benzothienyl, thiamorpholinyl, thiamorpholinyl sulfoxide, thiamorpholinyl sulfone, dioxaphospholanyl, oxadiazolyl and l,4-thiazine-l,l- dione. 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 "bridged heterocyclyl" refers to unsaturated or saturated bridged cyclic system that is not aromatic. Such a system may contain isolated or conjugated unsaturation, but not aromatic or heteroaromatic rings in its core structure (but may have aromatic substitution thereon). And at least one ring in the system is inclusive of one or more heteroatoms, wherein each ring in the system contains 3 to 7 ring members, e.g., 1 to 10 carbon atoms and 1 to 3 heteroatoms selected from N, O, P or S, wherein the S or P is optionally substituted with one or more oxo to provide the group SO or S0₂, PO or P0₂. Some non-limiting examples of bridged heterobicyclic ring system include (lR,5S)-8- azabicyclo[3.2.1]octane, (lR,5S)-3-azabicyclo[3.1.0]hexane, (S)-hexahydropyrrolo[l,2- a]pyrazin-4(lH)-one, (S)-octahydro-4H-pyrido[l,2-a]pyrazin-4-one or (lR,5S)-3,8- diazabicyclo[3.2.1]octane.

The term "fused bicycle" refers to a fused cycloalkane ring system wherein each ring has from 4 to 8 carbon atoms (i.e. C₈-Ci₆ fused bicyclic) and the fused ring system has from 0 to 3 bridge head carbon atoms. One or both of the fused rings may contain zero or one double bond. Examples of fused bicyclics include but are not limited to bicyclo[2,2,l]heptyl, bicyclo[2,2,l]heptenyl.

The term "heteroaryl" refers to an aromatic heterocyclic ring radical. Examples of such heteroaryl include, but are not limited to pyridyl, pyrazinyl, pyrimidinyl, oxazolyl, furanyl, quinolinyl, tetrazoyl, triazolyl, 1,3-Diaza-lH-indenyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, pyrazolo[l,5-a]pyrimidinyl, 1,3,4-Oxadiazolyl, thiazolidinyl, isothiazolyl, quinuclidinyl, isothiazolidinyl, indolyl, isoindolyl, indolinyl and isoindolinyl. 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.

"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" means a compound that is transformed in vivo to yield a compound of formula (I) or a pharmaceutically acceptable salt, hydrate or solvate, or metabolite of the compound. The transformation may occur by various mechanisms, such as through hydrolysis in blood. 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).

A "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, lxlO⁶ 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 IN 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 for 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-lymphotropic 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-lymphocytes 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, Antimicrobial 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. 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"), ³H, ⁿC, ¹³C, ¹⁴C, ¹³N, ¹⁵N, ¹⁵0, ¹⁷0, ¹⁸0, ³²P, ³³P, ³⁵S, ¹⁸F, ³⁶C1, ¹²³I and ¹²⁵I. Particular isotopes are -CD₃ or -C(D₂)-. 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 2010, 75, 1285-1288; Journal of organic chemistry 2000, 65, 3934-3940; Tetrahedron: asymmetry 2008, 19, 302-308; or Tetrahedron: asymmetry 2003, 14, 217-223.

Scheme- 1

[where in Ri is benzoic _acid, R₃ & R₄ together cyclopropyl, R2 & R5 is H, X is absent]

Compounds of formula (I) (wherein, Rf„ R₇, Rg, R9 and Y are same as defined above) can be prepared as described in Scheme 1. The cyclopropyl compounds of formula 1 (prepared as described in patent publication WO 2013/160810 and Pi & P₂ are protecting groups such as acetyl, benzyl or the like) can be converted to give the C3-hydroxy compounds of formula 2 in the presence of suitable bases such as potassium carbonate (K₂CO₃), sodium carbonate (Na₂C0₃), lithium hydroxide (LiOH), cesium carbonate (CS₂CO₃) or the like in the solvents such as methanol (MeOH), tetrahydrofuran (THF), acetone or the like. The C3-hydroxy compounds of formula 2 can be converted to give the C3-keto compounds of formula 3 in the presence of oxidising agents such as pyridinium chlorochromate (PCC), oxalyl chloride (COCl₂), Dess-martin reagent or the like in the solvents such as dimethyl sulfoxide (DMSO), dichloromethane (DCM), tetrahydrofuran (THF), acetone or the like, in presence of weak base such as triethylamine (TEA), N,N- diisopropylethylamine (DIPEA) or the like. The C3-keto compounds of formula 3 can be converted to give the C3-triflate compounds of formula 4 in the presence of N-phenyl-O- ((trifluoromethyl)sulfonyl)-N-(((trifluoromethyl)sulfonyl)oxy)hydroxylamine or the like in the presence of a base such as potassium bis(trimethylsilyl)amide (KHMDS) or the like in the solvents such as THF or the like. The C3-triflate compounds of formula 4 can be converted to give the C3-aryl compounds of formula 6 accomplished via Suzuki coupling with the corresponding boronic acid compounds of formula 5 in the presence of palladium (Pd) catalyst in the solvents such as 1,4-dioxane, propanol, water or the like in presence of a base such as sodium carbonate (Na₂C0₃), potassium carbonate (K₂C0₃) or the like. The selective deprotection and protection of the ester compounds of formula 6 in the C-28 position can be converted to give the C28-silylated compounds of formula 7 in the presence of tert-butyl dimethyl silane, palladium acetate (Pd(OAc)₂) or the like in the solvents such as dichloromethane (DCM), tetrahydrofuran (THF) or the like in the presence of a weak base such as triethylamine (TEA) or the like. The C28-silylated compounds of formula 7 can be deprotected in the presence of tetra-n-butyl ammonium fluoride (TBAF) or the like in the solvents such as 1,4-dioxane or the like affords the corresponding C28-carboxylic acid compounds of formula 8 which is converted to the acid chloride compounds of formula 9 in the presence of acid chlorides such as thionyl chloride (SOCl₂), oxalyl chloride (COCl₂) or the like in the solvents such as toluene or the like. The acid chloride compounds of formula 9 can be reacted with the amine compounds of formula 10 to provide the C-28 amide compounds of formula 11 in the presence of solvents such as dichloromethane (DCM), tetrahydrofuran (THF), acetone or the like in presence of a weak base such as triethylamine (TEA), Ν,Ν-Diisopropylethylamine (DIPEA) or the like. The C3-benzoic ester compounds of formula 11 can be deprotected to give the final C3-acid compounds of formula (I) in presence of a base such as lithium hydroxide (LiOH), lithium diisopropyl amide (LDA), potassium tert-butoxide (t-BuOK) or the like in the solvents such as tetrahydrofuran (THF), water 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-d₆ (Deuterated DMSO); d (Doublet); MeOH (Methanol); EtOAc (Ethyl acetate); g or gr (gram); H or H₂ (Hydrogen); HC1 (Hydrochloric acid); h or hr. (Hours); 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); N₂ (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)₂0 (Di-tert-butyl dicarbonate); DCM (dichloromethane); eq (equivalent); Ltr or L (Liter); CDC1₃ (Deuterated chloroform); J (Coupling constant); K₂C0₃ (potassium carbonate); Cs₂C0₃ (Cesium carbonate); NaHC0₃ (Sodium bicarbonate); Na₂S0₄ (Sodium sulphate); Pd/C (palladium in carbon) and Pd(OAc)₂ (palladium acetate); LiOH (lithium hydroxide); LDA (lithium diisopropyl amide); SOCl₂ (thionyl chloride); COCl₂ (oxalyl chloride); TBAF (tetra-n-butyl ammonium fluoride); Na₂C0₃ (sodium carbonate); KHMDS (potassium bis(trimethylsilyl)amide) .

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 them 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.

EXAMPLES

Example 1: Preparation of 4-((lR,3aS,5aR,5bR,7aR,l laS,l lbR,13aR,13bR)-5a,5b,8,8,l la- pentamethyl-l-(l-methylcvclopropyl)-3a-((S)-2-(5-phenyl-lH-imidazol-2-yl)pyrrolidine-l- carbonyl)-2,3,3a,4,5,5a,5b,6,7,7a,8,l l,l la,l lb,12,13,13a,13b-octadecahvdro-lH- cyclopentaral chrysen-9-yl)benzoic acid:

Step 1: Synthesis of benzyl (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-9-hydroxy- 5a,5b, 8,8,1 la-pentamethyl-1 -( 1 -methylcyclopropyl)icosahydro-3aH-cyclopenta[a Jchrysene- 3a-carboxylate:

To a stirred solution of benzyl (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9- acetoxy-5a,5b,8,8, 1 la-pentamethyl- l-(l-methylcyclopropyl)icosahydro-3aH-cyclopenta[a] chrysene-3a-carboxylate (prepared as described in WO 2013/160810 A2, 10 g, 16.6 mmol, 1.0 eq) in THF (100 ml) in Methanol (100 ml) was added potassium carbonate (16.0 g, 116.2 mmol, 7.0 eq). The reaction mixture was stirred at room temperature for 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 CH₂CI₂. The filtrate was evaporated under reduced pressure and the crude was purified by column chromatography by using 30% ethyl acetate and hexane as an eluent gave the compound (9.0 g, yield: 96.0%) as a white solid. H¹ NMR (CDCI₃, 300 MHz): δ 7.33 (m, 5H), 5.13-5.02 (q, 2H), 3.20-3.17 (t, 1H), 2.26-2.22 (m, 1H), 2.10-1.99 (m, 1H), 1.93-1.73 (m, 3H), 1.72-1.60 (m, 5H), 1.48-1.44 (m, 3H), 1.40-1.16 (m, 9H), 1.10-1.06 (m, 2H), 0.96 (s, 6H), 0.89 (s, 4H), 0.81 (s, 3H), 0.75 (s, 6H), 0.69-0.66 (m, 1H), 0.40-0.30 (m, 2H) and 0.26-0.17 (m, 2H); Mass: [M+H]⁺ 561.10 (100%).

Step 2: Synthesis of benzyl (lR,3aS,5aR,5bR,7aR,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla- pentamethyl-l-(l-methylcyclopropyl)-9-oxoicosahydro-3aH-cyclopenta[a]chrysene-3a- carboxylate:

The oxalyl chloride (2.02 mL, 24.10 mmol, 1.5 eq) was dissolved in DCM (40 mL). The resulting mixture was cooled to -78 °C then added DMSO (3.4 mL, 48.2 mmol, 3.0 eq) in DCM (40 mL) into the mixture under nitrogen atmosphere and stirred for 45 minutes. Then added benzyl (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy-5a,5b,8,8,l la- pentamethyl-l-(l-methylcyclopropyl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxylate (step 1, 9.0 g, 16.07 mmol, 1.0 eq) in DCM (100 mL). The mixture was stirred for 2 hours at -78 °C. Triethylamine (11.18 mL, 80.35 mmol, 5.0 eq) was added into the mixture drop wise. The resulting mixture was stirred for 1 hour at -78 °C and 2 hours at room temperature. The resulting mixture was diluted with ethyl acetate and washed with brine. The organic layer was dried and concentrated. The crude was purified by column chromatography by using 10% ethyl acetate and hexane as an eluent gave the desired compound (7.0 g, yield: 78.0%) as a white solid. H¹ NMR (CDC1₃, 300 MHz): δ 7.35-7.32 (m, 5H), 5.13-5.02 (q, 2H), 2.50- 2.40 (m, 2H), 2.28 (d, 1H), 2.41-2.10 (m, 1H), 2.00-1.80 (m, 3H), 1.71-1.62 (m, 3H), 1.46- 1.12 (m, 15H), 1.06 (s, 3H), 1.02 (s, 3H), 0.96 (s, 3H), 0.91-0.90 (m, 6H), 0.79 (s, 3H), 0.38- 0.32 (m, 2H) and 0.31-0.20 (m, 2H); Mass: [M+H]⁺ 558.20 (100%).

Step 3: Synthesis of benzyl (lR,3aS,5aR,5bR,7aR,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla- pentamethyl-1 -( 1 -methylcyclopropyl)-9-( ( ( trifluoromethyl )sulfonyl )oxy )-l,2, 3,4,5, 5a,5b, 6, 7, 7a, 8,ll,lla,llb,12,13,13a,13b-octadecahydro-3aH-cyclopenta[a]chrysene-3a-carboxylate:

To the stirred solution of benzyl (lR,3aS,5aR,5bR,7aR,l laR,l lbR,13aR,13bR)-

5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclopropyl)-9-oxoicosahydro-3aH-cyclopenta[a] chrysene-3a-carboxylate (step 2, 7.0 g, 12.54 mmol, 1.0 eq) in THF (50 mL) was added KHMDS (75 mL, 25.08 mmol, 2.0 eq) at -78 °C under nitrogen atmosphere and stirred for about 30 minutes, then added N-phenyl-Bis(trifluoromethane)sulfonamide (8.97 g, 37.63 mmol, 3.0 eq) in THF (20 mL) stirred at same temperature for 4 hours. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was quenched with saturated NaCl solution and extracted with DCM. The organic layer was dried with Na₂S0₄ and concentrated 30°C below water bath temperature. The crude was purified by column chromatography by using 2% ethyl acetate and hexane as an eluent give the compound (6.0 g, yield: 69.7%) as a white solid. H¹ NMR (CDC1₃, 300 MHz): δ 7.59- 7.52 (m, 3H), 7.49 (d, 2H), 7.39-7.33 (m, 4H), 5.58 (d, 1H), 5.55-5.02 (q, 2H), 2.26-2.01 (m, 3H), 1.97-1.90 (m, 1H), 1.88-1.61 (m, 4H), 1.50-1.21 (m, 8H), 1.1 (s, 4H), 1.01 (s, 3H), 0.97 (s, 3H), 0.96-0.85 (m, 8H), 0.78 (s, 3H), 0.40-0.31 (m, 2H) and 0.27-0.19 (m, 2H); Mass: [M+H]⁺ 690.29 (100%).

Step 4: Synthesis of benzyl (lR,3aS,5aR,5bR, 7aR,llaS,llbR,13aR,13bR)-9-(4-(methoxy carbonyl)phenyl)-5a,5b,8,8,lla-pentamethyl-l-(l-methylcyclopropyl)-l,2,3,4,5,5a,5b,6, 7,7a, 8,ll,lla,llb,12,13,13a,13b-octadecahydro-3aH-cyclopenta[a]chrysene-3a-carboxylate:

To stirred solution of benzyl (lR,3aS,5aR,5bR,7aR,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,1 la-pentamethyl-l-(l-methylcyclopropyl)-9-(((trifluoromethyl)sulfonyl)oxy)- 1,2,3,4,5, 5a,5b,6,7,7a,8,l 1,1 la,l lb, 12,13, 13a,13b-octadecahydro-3aH-cyclopenta[a]chrysene-3a- carboxylate (step 3, 2.5 g, 3.62 mmol, 1.0 eq ) in 1,4-dioxane (12.5 mL), 2-Propanol (12.5 mL) and water (7.5 mL) was added K₃P0₄ and 4-(methoxy carbonyl) benzene boronic acid (0.98 g, 5.43 mmol, 1.5 eq) at room temperature under nitrogen atmosphere then tetrakiss (triphenylphosphine) palladium (0.25 g, 0.2 mmol, 0.06 eq) were added under nitrogen atmosphere. The mixture was then heated to 90 °C and stirred for overnight. 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 CH₂CI₂. The filtrate was evaporated under reduced pressure and the crude was purified by column chromatography by using 4% ethyl acetate and hexane as an eluent give the desired product (1.0 g, yield: 41.6%) as a white solid. H¹ NMR (CDC1₃, 300 MHz): δ 7.94 (d, 2H), 7.35 (s, 5H), 7.21 (d, 2H), 5.30 (d, 1H), 5.15-5.07 (q, 2H), 3.90 (s, 3H), 2.27-2.09 (m, 2H), 2.23-2.10 (m, 1H), 2.00 (bs, 1H), 1.83- 1.76 (m, 3H), 1.52-1.33 (m, 9H), 1.29-1.22 (m, 5H), 1.19-1.11 (m, 3H), 0.99-0.97 (m, 5H), 0.92-0.88 (m, 9H), 0.85-082 (m, 3H), 0.41-0.32 (m, 2H) and 0.27-0.18 (m, 2H); Mass: [M+H]⁺ 676.19 (100%).

Step 5: Synthesis of tert-butyldimethylsilyl (lR,3aS,5aR,5bR, 7aR,llaS,llbR,13aR,13bR)-9- (4-(methoxycarbonyl)phenyl)-5a,5b,8,8,lla-pentamethyl-l-(l-methylcyclopropyl)-l,2,3,4,5, 5a,5b,6,7, 7a,8, 11,11 a,l lb, 12, 13,13a,13b-octadecahydro-3aH-cyclopenta[a]chrysene-3a- carboxylate:

To the stirred solution of benzyl (lR,3aS,5aR,5bR,7aR,l laS,l lbR,13aR,13bR)-9-(4- (methoxycarbonyl)phenyl)-5a,5b, 8, 8, l la-pentamethyl-l-(l-methylcyclopropyl)- 1,2,3,4,5, 5a, 5b,6,7,7a,8,l 1,1 la,l lb, 12,13, 13a,13b-octadecahydro-3aH-cyclopenta[a]chrysene-3a- carboxylate (step 4, 1.0 g, 1.47 mmol, 1.0 eq) in DCM (15 mL) was added Triethylamine (0.32 mL, 2.35 mmol, 1.6 eq) and tert. Butyl dimethylsilane (0.47 mL, 2.95 mmol, 2.0 eq) at room temperature under nitrogen atmosphere, then Pd(OAc)₂ (0.16 g, 7.35 mmol, 0.5 eq) was added to reaction mixture. Then the reaction was heated to 60 °C and stirred for about 4 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 CH₂C1₂. The filtrate was evaporated under reduced pressure, the crude was dissolved in acetic acid (5.2 mL), THF (2.6 mL), water (1.3 mL) and stirred for about 1 hour at room temperature. Solid was separated, filtered and washed with water to give the desired product (1.0 g, yield: 98.0%) as an off white solid. H¹ NMR (CDC1₃, 300 MHz): δ 7.94 (d, 2H), 7.21 (d, 2H), 5.31 (d, 1H), 3.91 (s, 3H), 2.25-2.09 (m, 3H), 2.23-2.10 (m, 2H), 2.00 (bs, 1H), 1.83-1.76 (m, 1H), 1.68-1.63 (m, 2H), 1.56 (ds, 4H), 1.51-1.43 (m, 4), 1.36-1.25 (m, 8H), 1.01-0.85 (m, 28H), 0.40-0.31 (m, 2H), 0.26 (s, 6H) and 0.06 (s, 2H); Mass: [M+H]⁺ 700.10 (100%).

Step 6: Synthesis of (lR,3aS,5aR,5bR, 7aR,llaS,llbR,13aR,13bR)-9-(4-(methoxycarbonyl) phenyl)-5a,5b,8,8,lla-pentamethyl-l-(l -methylcyclopropyl)-l,2,3,4,5,5a,5b,6, 7,7a,8,ll,lla, llb,12,13,13a,13b-octadecahydro-3aH-cyclopenta[a]chrysene-3a-carboxylic acid:

The stirred solution of tert-butyldimethylsilyl (lR,3aS,5aR,5bR,7aR,l laS,l lbR, 13aR,13bR)-9-(4-(methoxycarbonyl)phenyl)-5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclo propyl)-l,2,3,4,5,5a,5b,6,7,7a,8,l 1,1 la,l lb, 12,13, 13a,13b-octadecahydro-3aH-cyclopenta[a] chrysene-3a-carboxylate (step 5, 1.0 g, 1.42 mmol, 1.0 eq) in 1,4-dioxane was added TBAF (1.04 mL, 2.85 mmol, 2.0 eq) at room temperature and stirred for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was diluted with water stirred for about 30 minutes. Solid was separated, filtered and dried under vacuum to give the desired product (0.7 g, yield: 84.0%) as an off white solid. H¹ NMR (DMSO-d₆, 300 MHz): δ 11.94 (bs, 1H), 7.89 (d, 2H), 7.26 (d, 2H), 5.26 (d, 1H), 3.84 (s, 3H), 2.25-2.09 (m, 3H), 1.96 (bs, 1H), 1.78-1.58 (m, 5H), 1.51-1.33 (m, 5H), 1.29-1.12 (m, 6H), 0.99-0.94 (m, 9H), 0.89-0.85 (m, 12H), 0.34 (t, 2H) and 0.22 (t, 2H); Mass: [M+H]⁺ 586.10 (100%).

Step 7: Synthesis of methyl 4-((lR,3aS,5aR,5bR,7aR,llaS,llbR,13aR,13bR)-3a-(chloro carbonyl)-5a,5b,8,8,lla-pentamethyl-l-(l-methylcyclopropyl)-2,3,3a,4,5,5a,5b,6,7, 7a,8,ll, lla,llb,12,13,13a,13b-octadecahydro-lH-cyclopenta[a]chrysen-9-yl)benzoate:

To a stirred solution of (lR,3aS,5aR,5bR,7aR,l laS,l lbR,13aR,13bR)-9-(4-(methoxy carbonyl)phenyl)-5a,5b,8 ,8, 1 la-pentamethyl- 1 -( 1 -methylcyclopropyl)- 1 ,2,3 ,4,5, 5a,5b, 6,7,7a, 8, 11, 11 a, l ib, 12,13, 13 a, 13b-octadecahydro-3 aH-cyclopenta[a] chry sene-3 a-carboxylic acid (step 6, 0.450 g, 0.76 mmol, 1.0 eq) in toluene (4.5 ml) at 0 °C was added thionyl chloride (0.304 ml, 3.8 mmol, 5.0 eq). The reaction mixture was allowed to reach room temperature and heated to reflux for about 3 hours. After completion of the reaction monitored by TLC, the solvent was evaporated in vacuo providing a crude residue that was re-dissolved in toluene (2x10 ml) to remove the excess thionyl chloride. The solution was concentrated in vacuo to produce the desired product (0.450 g, yield: 98.0%) which is used as such for next step.

Step 8: Synthesis of methyl 4-((lR,3aS,5aR,5bR, 7aR,llaS,llbR,13aR,13bR)-5a,5b,8,8,lla- pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-(5-phenyl-lH-imidazol-2-yl)pyrrolidine-l- carbonyl)-2, 3, 3 a, 4, 5, 5 a, 5 b, 6,7, 7a,8,ll,lla,llb,12,13,l 3 a, 13b-octadecahydro-l H-cyclopenta [a] chrysen-9-yl)benzoate :

To a stirred solution of 4-phenyl-2-(pyrrolidin-2-yl)-lH-imidazole (prepared as described in WO 2014/105926 Al, 0.317 mg, 1.49 mmol, 2.0 eq) and triethylamine (0.516 niL, 3.7 mmol, 5.0 eq) in CH₂CI₂ (5 ml) at 0 °C was added a solution of methyl 4-((lR,3aS, 5aR,5bR,7aR,l laS,l lbR,13aR,13bR)-3a-(chlorocarbonyl)-5a,5b,8,8,l la-pentamethyl-l-(l- methylcyclopropyl)-2,3,3a,4,5,5a,5b,6,7,7a,8,l 1,1 la,l lb, 12,13, 13a,13b-octadecahydro-lH- cyclopenta [a]chrysen-9-yl)benzoate (step 7, 0.450 g, 0.745mmol, 1.0 eq) in DCM (5 ml). The reaction mixture was allowed to stir at room temperature for overnight. The reaction mixture was diluted with water and extracted with CH₂CI₂ (3x100 ml). The combined organic extracts were dried over Na₂S0₄, filtered and evaporated under reduced pressure. The residue was purified by column chromatography by using 20% ethyl acetate and hexane as an eluent gave the desired compound (0.5 g, yield: 86.0%) as a white solid. H¹ NMR (DMSO-d₆, 300 MHz): δ 11.54 (bs, 1H), 7.89 (d, 2H), 7.72 (d, 2H), 7.40-7.13 (m, 5H), 5.26 (d, 1H), 5.24 (s, 1H), 3.84 (s, 2H), 3.37 (bs, 1H), 3.64 (s, 1H), 2.81(d, 1H), 2.33 (d, 1H), 2.14- 2.20 (m, 4H), 1.86 (m, 3H), 1.86-1.42 (m, 12H), 1.29-1.23 (m, 9H), 0.95-0.86 (m, 17H), 0.27 (t, 2H) and 0.14 (t, 2H); Mass: [M+H]⁺ 782.12 (100%).

Step 9: Synthesis of 4-((lR,3aS,5aR,5bR, 7aR,llaS,llbR,13aR,13bR)-5a,5b,8,8,lla-penta methyl-1 -(1 -methylcyclopropyl)-3a-((S)-2-(5-phenyl-lH-imidazol-2-yl)pyrrolidine-l- carbonyl)-2, 3, 3 a, 4, 5, 5 a, 5 b, 6,7, 7a,8,ll,lla,llb,12,13,l 3 a, 13b-octadecahydro-l H-cyclopenta [a] chrysen-9-yl)benzoic acid:

To a stirred solution of methyl 4-((lR,3aS,5aR,5bR,7aR,l laS,l lbR,13aR,13bR)- 5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-(5-phenyl-lH-imidazol-2-yl) pyrrolidine-l-carbonyl)-2,3,3a,4,5,5a,5b,6,7,7a,8,l 1,1 la,l lb, 12,13, 13a,13b-octadecahydro- lH-cyclopenta[a] chrysen-9-yl)benzoate (step 8, 0.500 g, 0.64 mmol, 1.0 eq) in 1,4-Dioxane (10 ml) and water (10 ml) was added lithium hydroxide monohydried (0.188 g, 4.48 mmol, 7.0 eq) at room temperature. Then the reaction mixture was stirred for about 4 hours. TLC indicated starting material was consumed and the desired product was observed. Then the reaction was evaporated directly under vacuum to give residue. The residue was diluted with water and acidified with IN HC1 p was adjusted to 4. Then solid compound was separated, filtered, washed with water and dried under vacuum to give the pure compound (0.400 g, yield: 81.0%) as a white solid. H¹ NMR (DMSO-d₆, 300 MHz): δ 14.35 (bs, 1H), 12.75 (bs, 1H), 7.87-7.76 (m, 4H), 7.49-7.20 (m, 5H), 5.25 (d, 1H), 5.14 (s, 1H), 3.88 (bs, 1H), 3.65 (bs, 1H), 2.66-2.61 (m, 1H), 2.42-2.29 (m, 1H), 2.18-2.05 (m, 2H), 1.88-1.67 (m, 4H), 1.46-1.23 (m, 19H), 0.98-0.86 (m, 19H), 0.28 (bd, 2H) and 0.15 (bd, 2H); Mass: [M+H]⁺ 768.09 (100%); HPLC: 95.02%. Example 2: Preparation of 4-((lR,3aS,5aR,5bR,7aR,l laS,l lbR,13aR,13bR)-3a-(((lR,3S)- 2,2-dimethyl-3-(piperidine- l-carbonyl)cvclobutyl)carbamoyl)-5a,5b,8,8 J 1 a-pentamethyl- 1 - (l-methylcvclopropyn-2.3.3a.4.5.5a.5b.6.7.7a.8.11.11a.l lb.l2.13.13a.l3b-octadecahvdro- lH-cvclopentaralchrysen-9-yl)benzoic acid:

Step 1: Synthesis of methyl 4-((lR,3aS,5aR,5bR,7aR,llaS,llbR,13aR,13bR)-3a-(((lR,3S)- 2,2-dimethyl-3-(piperidine-l -carbonyl)cyclobut l)carbamoyl)-5a,5b,8,8,l 1 a-pentamethyl-1 - (l-methylcyclopropyl)-2,3,3a,4,5,5a,5b,6,7, 7a,8,ll,lla,llb,12,13,13a,13b-octadecahydro- lH-cyclopenta[a]chrysen-9-yl)benzoate:

To a stirred solution of (lS,3R)-3-amino-2,2-dimethylcyclobutyl)(piperidin-l-yl) methanone (prepared as described in WO 2013/160810 A2, 0.278 g, 1.32 mmol, 2.0 eq) and triethylamine (0.5 mL, 3.3 mmol, 5.0 eq) in CH₂CI₂ (5 ml) at 0 °C was added a solution of methyl 4-((lR,3aS,5aR,5bR,7aR,l laS,l lbR,13aR,13bR)-3a-(chlorocarbonyl)-5a,5b,8,8, 11a- pentamethyl-l-(l-methylcyclopropyl)-2,3,3a,4,5,5a,5b,6,7,7a,8,l 1,1 la,l lb, 12,13, 13a,13b- octadecahydro-lH-cyclopenta[a]chrysen-9-yl)benzoate (Example 1-step 7, 0.400 g, 0.662 mmol, 1.0 eq) in DCM (5 ml). The reaction mixture was allowed to stir at room temperature for overnight. The reaction mixture was diluted with water and extracted with CH₂CI₂ (3x100 ml). The combined organic extracts were dried over Na₂S0₄, filtered and evaporated under reduced pressure. The residue was purified by column chromatography by using 20% ethyl acetate and hexane as an eluent gave the desired compound (0.400 g, yield: 78.0%) as a white solid. H¹ NMR (CDCI3, 300 MHz): δ 7.93 (d, 2H), 7.21 (d, 2H), 5.85 (d, 1H), 5.31 (d, 1H), 4.13-4.05 (q, 1H), 3.90 (s, 3H), 3.70 (bd, 1H), 3.45 (t, 3H), 2.89-2.84 (t, 1H), 2.49-2.23 (m, 3H), 2.04 (m, 2H), 1.91-1.71 (m, 6H), 1.42 (s, 11H), 1.37 (d, 8H), 1.01-0.86 (m, 25H), 0.44-0.26 (m, 2H) and 0.24-0.07 (m, 2H); Mass: [M+H]⁺ 778.09 (100%).

Step 2: Synthesis of 4-((lR,3aS,5aR,5bR, 7aR,llaS,llbR,13aR,13bR)-3a-(((lR,3S)-2,2- dimethyl-3-(piperidine-l-carbonyl)cyclobut l)carbamoyl)-5a,5b,8,8,lla-pentamethyl-l-(l- methylcyclopropyl)-2,3,3a,4,5,5a,5b,6, 7,7a,8,ll,lla,llb,12,13,13a,13b-octadecahydro-lH- cyclopenta[a chrysen-9-yl)benzoic acid:

To a stirred solution of methyl 4-((lR,3aS,5aR,5bR,7aR,l laS,l lbR,13aR,13bR)-3a- (((lR,3S)-2,2-dimethyl-3-(piperidine-l-carbonyl)cyclobutyl)carbamoyl)-5a,5b,8,8,l la- pentamethyl-l-(l-methylcyclopropyl)-2,3,3a,4,5,5a,5b,6,7,7a,8,l 1,1 la,l lb, 12,13, 13a,13b- octadecahydro-lH-cyclopenta[a]chrysen-9-yl)benzoate (step 1, 0.400 g, 0.514 mmol, 1.0 eq) in 1,4-Dioxane (10 ml) and water (10 ml) was added Lithium hydroxide monohydried (0.151 g, 3.59 mmol, 7.0 eq) at room temperature. Then the reaction mixture was stirred for 4 hours. TLC indicated starting material was consumed and the desired product was observed. Then the reaction was evaporated directly under vacuum to give residue. The residue was diluted with water and acidified with IN HC1 p was adjusted to 4. Then solid compound was separated which solid was filtered and washed with water and dried under vacuum to give the pure compound (0.350 g, yield: 89.0%) as a white solid. H¹ NMR (DMSO-d₆, 300 MHz): δ 12.79 (bs, 1H), 7.87 (d, 2H), 7.48 (d, 1H), 7.23 (d, 2H), 5.25 (d, 1H), 3.97 (t, 1H), 3.56 (s, 2H), 2.90-2.84 (t, 1H), 2.20-2.07 (m, 3H), 1.93-2.22 (m, 20H), 1.0-1.04 (m, 11H), 0.96-0.88 (m, 20H), 0.73 (s, 3H), 0.31 (bd, 2H) and 0.19 (bd, 2H); Mass: [M+H]⁺ 765.09 (100%); HPLC: 95.67%.

Example 3: Preparation of 4-((lR,3aS,5aR,5bR,7aR,l laS,l lbR,13aR,13bR)-3a-(dimethyl carbamoyr)-5a,5b,8,8 J la-pentamethyl-l-(l-methylcvclopropyl)-2,3,3a,4,5,5a,5b,6,7 ,7a,8 ,11, 11 a, l ib, 12, 13, 13a, 13b-octadecahydro- 1 H-cyclopentaralchrysen-9-yl)benzoic acid:

Step 1: Synthesis of methyl 4-((lR,3aS,5aR,5bR, 7aR,llaS,llbR,13aR,13bR)-3a-(dimethyl carbamoyl)-5a,5b,8,8,lla-pentamethyl-l-(l-methylcyclopropyl)-2,3,3a,4,5,5a,5b,6,7, 7a,8,ll, lla,llb,12,13,13a,13b-octadecahydro-lH-cyclopenta[a]chrysen-9-yl)benzoate:

To a stirred solution of dimethylamine (0.87 mL, 1.65 mmol, 2.0 eq) and triethylamine (0.57 mL, 4.13 mmol, 5.0 eq) in CH₂CI₂ (10 ml), a solution of methyl 4-((lR, 3aS,5aR,5bR,7aR,l laS,l lbR,13aR,13bR)-3a-(chlorocarbonyl)-5a,5b,8,8,l la-pentamethyl-l- (l-methylcyclopropyl)-2,3,3a,4,5,5a,5b,6,7,7a,8,l 1,1 la,l lb, 12,13, 13a,13b-octadecahydro-lH -cyclopenta[a]chrysen-9-yl)benzoate (Example 1-step 7, 0.500 g, 0.662 mmol, 1.0 eq) in DCM (10 ml) was added at 0 °C. The reaction mixture was allowed to stir at room temperature for overnight. The reaction mixture was diluted with water and extracted with CH₂CI₂ (3x100 ml). The combined organic extracts were dried over Na₂S0₄, filtered and evaporated under reduced pressure. The residue was purified by column chromatography by using 20% ethylacetate and hexane as an eluent to afford the desired compound (0.400 g, yield: 80.0%) as a white solid. H¹ NMR (CDCI₃, 300 MHz): δ 7.94 (d, 2H), 7.21 (d, 2H), 5.31 (d, 1H), 3.90 (s, 3H), 2.97 (s, 6H), 2.88-2.79 (m, 1H), 2.25-1.99 (m, 4H), 1.80-1.52 (m, 4H), 1.47-1.35 (m, 11H), 1.35-1.21 (m, 5H), 1.19-1.00 (m, 8H), 0.99-0.92 (m, 8H), 0.47-0.31 (m, 2H) and 0.27-0.15 (m, 2H); Mass: [M+H]⁺ 614.09 (100%).

Step 2: Synthesis of 4-((lR,3aS,5aR,5bR, 7aR,llaS,llbR,13aR,13bR)-3a-(dimethyl carbamoyl)-5a,5b,8,8,lla-pentamethyl-l-(l-methylcyclopropyl)-2,3,3a,4,5,5a,5b,6,7, 7a, 8,ll,lla,llb,12,13,13a,13b-octadecahydro-lH-cyclopenta[a] chrysen-9-yl)benzoic acid:

To a stirred solution of methyl 4-((lR,3aS,5aR,5bR,7aR,l laS,l lbR,13aR,13bR)-3a- (dimethylcarbamoyl)-5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclopropyl)-2,3,3a,4,5,5a,5b, 6,7,7a,8, 11 , 1 la, 1 lb, 12, 13, 13a, 13b-octadecahydro- lH-cyclopenta[a]chrysen-9-yl)benzoate (step 1, 0.400 g, 0.652 mmol, 1.0 eq) in THF (7 ml) and water (7 ml), lithium hydroxide monohydried (0.19 g, 4.56 mmol, 7.0 eq) was added at room temperature. Then the reaction mixture was stirred for overnight. TLC indicated starting material was consumed and the desired product was observed. Then the reaction was evaporated directly under vacuum to give residue. The residue was diluted with water and acidified with IN HC1 (p was adjusted to 4). Then solid compound was separated which solid was filtered, washed with water and hexane dried under vacuum to afford the desired pure compound (0.290 g, yield: 71.0%) as a white solid. H¹ NMR (DMSO-d₆, 300 MHz): δ 12.46 (s, 1H), 7.73 (d, 2H), 6.97 (d, 2H), 5.20 (d, 1H), 2.88 (s, 6H), 2.80-2.72 (m, 1H), 2.27-1.98 (m, 4H), 1.71-1.38 (m, 13H), 1.23-1.14 (m, 7H), 0.97 (d, 5H), 0.90 (t, 11H), 0.32 (m, 2H) and 0.20 (m, 2H); Mass: [M+H]⁺ 600.53 (100%); HPLC: 97.78%.

Example 4: Preparation of 4-(lR,3aS,5aR,5bR,7aR,l laS,l lbR,13aR,13bR)-5a,5b,8,8,l la- pentamethyl- 1 -( 1 -methylcyclopropyl)-3 a-((S )-2-(pyrrolidin- 1 -ylmethyDpyrrolidine- 1 - carbonyl)-2,3,3aA5,5a,5b,6J Ja,8 J lJ laJ lbJ2J3 J3aJ3b-octadecahvdro-lH-cvclopenta ralchrysen-9-vDbenzoic acid:

Step 1: Synthesis of methyl 4-((lR,3aS,5aR,5bR, 7aR,llaS,llbR,13aR,13bR)-5a,5b,8,8,lla- pentamethyl-1 -( 1 -methylcyclopropyl)-3a-( (S)-2-(pyrrolidin-l -ylmethyl)pyrrolidine-l - carbonyl)-2, 3, 3 a, 4, 5, 5 a, 5 b, 6,7, 7a,8,ll,lla,llb,12,13,l 3 a, 13b-octadecahydro-l H-cyclopenta [a]chrysen-9-yl)benzoate:

To a stirred solution of (S)-l-(pyrrolidin-2-ylmethyl)pyrrolidine (procured form Aldrich, 0.250 mL, 1.57 mmol, 1.5 eq) and triethylamine (0.72 mL, 5.2 mmol, 5.0 eq) in CH₂C1₂ (40 ml) at 0 °C was added a solution of methyl 4-((lR,3aS,5aR,5bR,7aR,l laS,l lbR, 13aR,13bR)-3a-(chlorocarbonyl)-5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclopropyl)-2,3,3a, 4,5,5a,5b,6,7,7a,8,l 1,1 la,l lb, 12,13, 13a,13b-octadecahydro-lH-cyclopenta[a]chrysen-9-yl) benzoate (Example 1-step 7, 0.600 g, 1.04 mmol, 1.0 eq) in DCM (5 ml). The reaction mixture was allowed to stir at room temperature for overnight. The reaction mixture was diluted with water and extracted with CH₂CI₂ (3x100 ml). The combined organic extracts were dried over Na₂S0₄, filtered and evaporated under reduced pressure. The residue was purified by column chromatography by using 4% MeOH and DCM as an eluent to afford the desired compound (0.600 g, yield: 85.0%) as a white solid. H¹ NMR (DMSO-d₆, 300 MHz): δ 7.87 (d, 2H), 7.23 (d, 2H), 5.26 (d, 1H), 4.19 (s, 1H), 3.84 (s, 3H), 3.51 (bs, 2H), 2.99-2.80 (m, 5H), 2.27-2.08 (m, 2H), 1.89-1.69 (m, 12H), 1.57-1.39 (m, 11H), 1.28-1.16 (m, 7H), 0.98-0.89 (m, 18H), 0.34 (m, 2H) and 0.21 (m, 2H); Mass: [M+H]⁺ 723.65 (100%).

Step 2: Synthesis of 4-(lR,3aS,5aR,5bR,7aR,llaS,llbR,13aR,13bR)-5a,5b,8,8,lla-penta methyl-l-(l -methylcyclopropyl)-3a-((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidine-l-carbonyl)-2, 3,3a,4,5,5a,5b,6,7, 7a,8,ll,lla,llb,12,13,13a,13b-octadecahydro-lH-cyclopenta[a]chrysen- 9-yl)benzoic acid:

To a stirred solution of methyl 4-((lR,3aS,5aR,5bR,7aR,l laS,l lbR,13aR,13bR)-

5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-(pyrrolidin-l-ylmethyl) pyrrolidine-l-carbonyl)-2,3,3a,4,5,5a,5b,6,7,7a,8,l 1,1 la,l lb, 12,13, 13a,13b-octadecahydro- lH-cyclopenta[a]chrysen-9-yl)benzoate (step 1, 0.600 g, 0.84 mmol, 1.0 eq) in THF (30 ml) and water (10 ml) was added lithium hydroxide monohydried (0.350 g, 8.4 mmol, 10.0 eq). The reaction mixture was refluxed for about 12 hours. TLC indicated starting material was consumed and the desired product was observed. Then the reaction mixture was evaporated directly under vacuum to give the residue. The residue was diluted with water and acidified with IN HC1 (p was adjusted to 4). The obtained solid compound was separated, filtered, washed with water and dried under vacuum to give the crude product which was purified by column chromatography by using 10% MeOH and DCM as an eluent to afford the desired compound (0.130 g, yield: 30.0%) as a white solid.H¹ NMR (DMSO-d₆, 300 MHz): δ 12.94 (bs, 1H), 7.87 (d, 2H), 7.23 (d, 2H), 5.26 (d, 1H), 4.19 (s, 1H), 3.51 (bs, 2H), 2.99-2.80 (m, 5H), 2.27-2.08 (m, 2H), 1.89-1.69 (m, 12H), 1.57-1.39 (m, 11H), 1.28-1.16 (m, 7H), 0.98- 0.89 (m, 18H), 0.34 (m, 2H) and 0.21 (m, 2H); Mass: [M+H]⁺ 709.55 (100%); HPLC: 97.55%.

Although the present application has been illustrated by certain of the preceding examples, it is not to be construed as being limited thereby; but rather, the present application encompasses the generic area as hereinbefore disclosed. Various modifications and embodiments can be made without departing from the spirit and scope thereof. For example, the following compounds are also included in the scope of the present application.

PHARMACOLOGICAL ACTIVITY

The compounds described herein can be tested for their antiviral 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 5: 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 C0₂ 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 IC₅₀ value of the example 4 is less than 50 nM & example 2 is more than 50 nM.

For 45 mg/ml HSA serum binding assay, wherein IC₅₀ value of the example 4 is less than 50 nM & example 2 is more than 50 nM.

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.

Claims

Claims:

1. A compound of the formula (I):

Formula (T)

wherein,

Ri is substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkadienyl,

substituted or unsubstituted aryl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted heteroaryl (such as, pyridine, pyrazine, pyrimidine, oxazole or thiazole), or substituted or unsubstituted fused bicycle; wherein R_b at each occurrence is hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl;

R₂ is hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted aminoacid, substituted or unsubstituted alkoxy, or substituted or unsubstituted cycloalkyl;

X is absent, O, S, CH₂ or NR_a; (wherein R_a is hydrogen, 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 taken together to form N-contained heterocyclyl (such as, pyrrolidine, piperdine, piperzine, or morpholine));

Y is absent, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heteroaryl, or -(CR_eRf)_m;

R₃ and R₄ are independently selected from hydrogen, hydroxy, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted alkoxylalkoxy, or substituted or unsubstituted aminoacid; wherein amino acid is substituted by substituted or unsubstituted alkyl; or R₃ and R₄ are taken together with their carbon atoms to which they are attached to form a bond; or R₃ and R₄ are taken together with their carbon atoms to which they are attached to form cycloalkyl; or R₃ and R₄ are taken together with the carbon atoms to which they are attached to form epoxide;

R5 and R_c are independently selected from hydrogen, C(0)₂R_d (wherein R_d is hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl), or substituted or unsubstituted alkyl;

R₆, R7 and R₈ are independently selected from hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl; or R₆ and R₇ are taken together with the nitrogen and carbon to which they are attached to form substituted or unsubstituted 4-10 membered heterocyclyl, or substituted or unsubstituted 5-15 membered bridged heterocyclyl; or R₇ and R₈ are taken together with the carbon atom to which they are attached to form 3-7 membered cycloalkyl or heterocyclyl;

R9 is absent, substituted or unsubstituted amine, substituted or unsubstituted 4-10 membered heterocyclyl, substituted or unsubstituted 4-10 membered heteroaryl, - C(0)heterocyclyl or substituted or unsubstituted aryl; wherein the substituent can be alkyl;

R_e and R_f are independently selected from hydrogen, or substituted or unsubstituted alkyl;

'n' is an integer from 0-1; and

'm' is an integer from 0-2;

or pharmaceutically acceptable salts, pharmaceutically acceptable solvates, pharmaceutically acceptable hydrates, tautomers, stereoisomers, prodrugs, compositions or combination thereof.

2. The compound of claim 1, wherein Ri is benzoic acid.

3. A compound selected from the group consisting of:

4-((lR,3aS,5aR,5bR,7aR,l laS,l lbR,13aR,13bR)-5a,5b,8,8,l la-pentamethyl-l-(l- methylcyclopropyl)-3a-((S)-2-(5-phenyl-lH-imidazol-2-yl)pyrrolidine-l-carbonyl)-2,3,3a, 4,5,5a,5b,6,7,7a,8,l 1,1 la,l lb, 12,13, 13a,13b-octadeca ydro-lH-cyclopenta[a]chrysen-9- yl)benzoic acid,

4-((lR,3aS,5aR,5bR,7aR,l laS,l lbR,13aR,13bR)-3a-(((lR,3S)-2,2-dimethyl-3- (piperidine-l-carbonyl)cyclobutyl)carbamoyl)-5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclo propyl)-2,3,3a,4,5,5a,5b,6,7,7a,8,l 1,1 la,l lb, 12,13, 13a,13b-octadecahydro-lH-cyclopenta[a] chrysen-9-yl)benzoic acid,

4-((lR,3aS,5aR,5bR,7aR,l laS,l lbR,13aR,13bR)-3a-(dimethylcarbamoyl)-5a,5b,8, 8,1 la-pentamethyl-l-(l-methylcyclopropyl)-2,3,3a,4,5,5a,5b,6,7,7a,8,l 1,1 la,l lb, 12,13, 13a, 13b-octadecahydro-lH-cyclopenta[a]chrysen-9-yl)benzoic acid, and

4-(lR,3aS,5aR,5bR,7aR,l laS,l lbR,13aR,13bR)-5a,5b,8,8,l la-pentamethyl-l-(l- methylcyclopropyl)-3a-((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidine-l-carbonyl)-2,3,3a,4,5,5a, 5b,6,7,7a,8,l 1,1 la,l lb, 12,13, 13a,13b-octadecahydro-lH-cyclopenta[a]chrysen-9-yl)benzoic acid,

or pharmaceutically acceptable salts, pharmaceutically acceptable solvates, hydrates, tautomers, stereoisomers, prodrugs, compositions or combination thereof.

4. A pharmaceutical composition comprising a compound according to any one of claims 1-3 and at least one pharmaceutically acceptable excipient.

5. The pharmaceutical composition according to claim 4, wherein the pharmaceutically acceptable excipient is a carrier or diluent.

6. A method for preventing, ameliorating or treating a viral mediated disease, disorder or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1-3.

7. The method according to claim 6, wherein the viral mediated disease, disorder or syndrome is HIV infection, HBV infection, HCV infection, a retroviral infection genetically related to AIDS, respiratory disorders (including adult respiratory distress syndrome (ARDS)), inflammatory disease, or a combination thereof.

8. A method of treating HIV in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1-3.

9. The method according to claim 6 and 8, wherein the subject is a mammal, including human.

10. Use of a compound according to any of claim 1 to 3, in the manufacture of a medicament for the treatment of viral mediated diseases.