WO2015198263A2 - Nouveaux dérivés substitués par proline bétulinique utilisés comme inhibiteurs du vih - Google Patents

Nouveaux dérivés substitués par proline bétulinique utilisés comme inhibiteurs du vih Download PDF

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WO2015198263A2
WO2015198263A2 PCT/IB2015/054784 IB2015054784W WO2015198263A2 WO 2015198263 A2 WO2015198263 A2 WO 2015198263A2 IB 2015054784 W IB2015054784 W IB 2015054784W WO 2015198263 A2 WO2015198263 A2 WO 2015198263A2
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carbonyl
pyrrolidine
lbr
lar
oxy
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WO2015198263A3 (fr
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Parthasaradhi Reddy BANDI
Rathnakar Reddy KURA
Panduranga Reddy ADULLA
David Krupadanam GAZULA LEVI
Eswara Rao BAMMIDI
Ranga Reddy KOTHAKAPU
Vl Subrahmanyam LANKA
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Hetero Research Foundation
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Hetero Research Foundation
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D451/00—Heterocyclic compounds containing 8-azabicyclo [3.2.1] octane, 9-azabicyclo [3.3.1] nonane, or 3-oxa-9-azatricyclo [3.3.1.0<2,4>] nonane ring systems, e.g. tropane or granatane alkaloids, scopolamine; Cyclic acetals thereof
    • C07D451/02—Heterocyclic compounds containing 8-azabicyclo [3.2.1] octane, 9-azabicyclo [3.3.1] nonane, or 3-oxa-9-azatricyclo [3.3.1.0<2,4>] nonane ring systems, e.g. tropane or granatane alkaloids, scopolamine; Cyclic acetals thereof containing not further condensed 8-azabicyclo [3.2.1] octane or 3-oxa-9-azatricyclo [3.3.1.0<2,4>] nonane ring systems, e.g. tropane; Cyclic acetals thereof
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/04—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings directly linked by a ring-member-to-ring-member bond

Definitions

  • the present invention relates to novel betulinic proline substituted derivatives and related compounds, compositions useful for therapeutic treatment of viral diseases and particularly HIV mediated diseases.
  • HIV Human Immunodeficiency Virus
  • AIDS Acquired Immunodeficiency Syndrome
  • 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.
  • 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. 199457(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.
  • WO 2014/105926 describes novel betulinic acid proline derivatives as HIV inhibitors
  • WO 2013/160810 describes novel betulinic acid derivatives as HIV inhibitors
  • Journal of Medicinal Chemistry (2012), 55(18), 8128-8136 describes Novel C-28 modified bevirimat analogues as potent HIV maturation inhibitors
  • 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 2011/007230 describes lupeol-type triterpene derivatives as antivirals
  • WO 2010/132334 describes 3, 28-disubstituted betulinic acid derivatives as anti- HIV agents
  • Journal of Medicinal Chemistry (2009), 52(10), 3248-3258 describes Design, Synthesis, Metabolic
  • 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
  • WO 2007/141392 describes cosmetic and pharmaceutical compositions comprising betulonic acid and betulin derivatives
  • Pharmaceutical Chemistry Journal, 2002, 36(9), 29-32 describes synthesis and anti-inflammatory activity of new acylated betulin derivatives.
  • the present invention relates to the compounds of the formula (1):
  • Ri can be substituted or unsubstituted alkyl, substituted or unsubstituted aryl,
  • R 2 can be H, 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 Ra can be H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, or Ra can be with their adjacent N and Carbon together form N-contained heterocycle (Preferably, pyrrolidine, piperdine, piperzine, or morpholine);
  • R 3 and R 4 can be independently selected from H, OH, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted alkoxylalkoxy, or substituted or unsubstituted aminoacids and preferably amino acids are substituted by substituted or unsubstituted alkyl, phosphoric acid, or phosphorus prodrugs or R 3 and R 4 can be together with their adjacent carbons to form a bond or R 3 and Rj can be together with their adjacent carbons to form cyclopropyl or R 3 and R 4 can be together with their adjacent carbons to form epoxide;
  • W can be C(O), C(S), or CR 7 R 8 ;
  • R x can be H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl and specifically heterocycles are imidazoles, oxazoles, oxadiazoles, triazoles, thiazoles, thiadizoles, isothiazoles, isothiadiazoles, pyridines, pyrazines, pyrimidines, or pyridazines;
  • R c can be hydrogen or substituted or unsubstituted alkyl
  • R 6 can be independently selected from H, hydroxy, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy;
  • R5, R 7 and R 8 can be independently selected from H, C0 2 Rd (wherein Rd can be H, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl), or substituted or unsubstituted alkyl;
  • n can be an integer from 1 to 3.
  • compositions of the formula (1) are also contemplated.
  • pharmaceutically acceptable solvates, including hydrates, of the compounds of the formula (1) are contemplated.
  • formula (1) structurally encompasses all stereoisomers, including enantiomers, diastereomers, racemates, and combinations thereof which may be contemplated from the chemical structure of the genus described herein.
  • formula (1) structurally encompasses all tautomers. Also contemplated are prodrugs of the compounds of the formula (1), including ester prodrugs.
  • a compound of formula (1) wherein Rx is substituted or unsubstituted oxadiazole, substituted or unsubstituted oxazole, substituted or unsubstituted imidazole and substituted or unsubstituted triazole. Most specifically the substituents are isopropyl, t-butyl, substituted or unsubstituted phenyl, pyridine, and thiophene. According to one embodiment, there is provided a compound of formula (1), wherein
  • R 2 is H.
  • W is C(O).
  • n 1
  • R b can be H, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl
  • R 2 can be H, substituted or unsubstituted alkyl, substituted or unsubstituted aminoacids, substituted or unsubstituted alkoxy or substituted or unsubstituted cycloalkyl;
  • X can be absent, O, S, CH 2i or NR a (wherein R a can be H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, or R a can be with their adjacent N and Carbon together form N-contained heterocycle (Preferably, pyrrolidine, piperdine, piperzine, or morpholine);
  • R 3 and R 4 can be independently selected from H, OH, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted alkoxylalkoxy, or substituted or unsubstituted aminoacids and preferably amino acids are substituted by substituted or unsubstituted alkyl, phosphoric acid, or phosphorus prodrugs or R 3 and R4 can be together with their adjacent carbons to form a bond or R 3 and Rj can be together with their adjacent carbons to form cyclopropyl or R 3 and R 4 can be together with their adjacent carbons to form epoxide;
  • W can be C(O), C(S), or CR 8 R 9 ;
  • R 6 can be H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl and most preferably alkyl is methyl, ethyl, isopropyl, and t-butyl, most preferably aryl is substituted or unsubstituted phenyl, most preferably heteroaryl is pyridine and R 6 is preferably substituted or unsubstituted isopropyl, t-butyl, phenyl, pyridine, pyrazine, pyrimidine or chromene;
  • R5, R 8 and R9 can be independently selected from H, C0 2 R d (wherein R d can be H, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl), or substituted or unsubstituted alkyl;
  • R 7 can be independently selected from H, hydroxy, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy;
  • 'n' can be an integer from 1 to 3.
  • compositions of the formula (1A) are also contemplated.
  • pharmaceutically acceptable solvates, including hydrates, of the compounds of the formula (1A) are 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).
  • 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 cause 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.
  • the present invention provides betulinic proline substituted 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.
  • 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), n-butyl, n-pentyl, and 1,1-dimethylethyl (t-butyl).
  • 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).
  • 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.
  • amino acid 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.
  • 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.
  • 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.
  • aryl refers to an aromatic radical having from 6 to 14 carbon atoms such as phenyl, naphthyl, tetrahydronapthyl, indanyl, and biphenyl.
  • halogen or halo includes fluorine, chlorine, bromine, or iodine.
  • heterocyclyl and “heterocyclic ring” refer to a stable 3- to 15-membered ring radical which consists of carbon atoms and from one to five heteroatoms selected from nitrogen, phosphorus, oxygen and sulfur.
  • 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.
  • the nitrogen atom may be optionally quaternized; and the ring radical may be partially or fully saturated (i.e., heterocyclic or heteroaryl).
  • 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, iso
  • 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.
  • prodrug means a compound that is transformed in vivo to yield a compound of Formula (1), (1A) 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.
  • treating or “treatment” of a state, disease, disorder or condition includes:
  • the benefit to a subject receiving treatment is either statistically significant or at least perceptible to the subject or to the physician.
  • 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).
  • domestic animals e.g., household pets including cats and dogs
  • 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 (1), or (1A), the present invention extends to these stereo isomeric forms and to mixtures thereof.
  • 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.
  • 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).
  • pharmaceutically acceptable excipient such as a pharmaceutically acceptable carrier or diluent.
  • contemplated pharmaceutical compositions include a compound(s) described herein in an amount sufficient to treat viral infection in a subject.
  • the subjects contemplated include, for example, a living cell and a mammal, including human.
  • 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.
  • 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.
  • 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.
  • 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).
  • 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.
  • 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.
  • injectable solutions or suspensions specifically aqueous solutions with the active compound dissolved in polyhydroxylated castor oil.
  • Antiviral HIV activity and cytotoxicity of compounds 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.
  • 20 ⁇ 1 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.
  • 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 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.
  • SAIC Fredrick P24 antigen capture assay kit
  • 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).
  • 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 ⁇ aliquots will be incubated with serial dilutions of compounds (50 ⁇ ) 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.
  • the present invention provides compounds and pharmaceutical formulations 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.
  • 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).
  • ARDS adult respiratory distress syndrome
  • bronchitis chronic bronchitis
  • chronic obstructive pulmonary disease cystic
  • the compounds described herein may be prepared by techniques known in the art.
  • 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.
  • 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.
  • R 6 is more specifically isopropyl, t-butyl, substituted or unsubstituted phenyl, pyridine, and thiophene
  • P can be a protecting group more specifically BOC, benzyl, MOM chloride (chloromethyl methyl ether), tosyl, TBDMS, p-methoxy phenyl (PMP), CBZ chloride, THP, dimethyl acetol, allyl ether, t-butyl ether or pthalimide can be prepared as described in Scheme 1.
  • the acetyl compounds of formula 1 can be converted to bromo compounds of formula 2 in the presence of bromine, N-bromosuccinimide, or the like in the solvents such as THF, diethyl ether, or the like.
  • the bromo compounds of formula 2 can be coupled with BOC protected proline compounds of formula 3 to get the compound of prolinated compounds of formula 4 in the presence of suitable coupling agents such as DIPEA, TEA, potassium carbonate or the like in the solvents such as N,N-Dimethylformamide, dichloromethane, dichloroethane, or the like.
  • the prolinated compounds of formula 4 can be cyclized to proline-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 proline-cyclic compounds (BOC protected) of formula 5 can be deprotected to give NH proline compounds of formula 6 in the presence of TFA, HC1 or the like in the solvents such as DCM, ethyl acetate or the like.
  • a base like triethyl amine, diisopropyl ethyl mine, or pyridine
  • an inert solvent like DCM, toluene, THF or a basic solvent like pyridine with or without addition of a catalyst like DMAP.
  • a C-3 alcohol of compounds of the formula 7 can be protected by an acetyl group in the presence of acetic anhydride (as described in T.W.Greene and P.G.M.Wuts, protective groups in organic synthesis, 3 edition, John Wiley & sons, New York, 1999) in the solvents such as pyridine or the like to give the C-3 acetyl compounds of formula 8.
  • acetic anhydride as described in T.W.Greene and P.G.M.Wuts, protective groups in organic synthesis, 3 edition, John Wiley & sons, New York, 1999
  • the C-3 acetyl (C-28 acid) compounds of formula 8 can be couple with substituted proline compounds of formula 9 (synthesized as described in Scheme 1) to give the C-28 cyclic amide compounds of formula 10 in the presence of oxylylchloride, TEA or the like in the presence of solvents such as DCM, or the like.
  • the C-28 cyclic amide (C-3 ester) compounds of formula 10 can be hydrolysed to give C-3 hydroxy compounds of formula 11 in the presence of bases such as potassium carbonate, sodium hydroxide, ammonia or the like in the solvents such as methanol:THF, methanol: water, 1,4-dioxane, methanol or the like.
  • Step 1 Synthesis of 2-bromo-l -(pyridin-2- ethanone: To the stirred solution of l-(pyridin-2-yl) ethanone (10 g, 82.6 mmol) in 120 ml of CC14 at 0°C was added bromine (4.2 ml, 26.2 mmol) dissolved in CC14 (20 ml) for about 15 min, and stirred for about 2 hrs at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was, concentrated and the crude product was dissolved in n-hexane and stirred for 30 min. The solid obtained was filtered and washed with ft-hexane, dried and proceeded for next step (wt 11.0 g).
  • Step 2 Synthesis of (S)-l-tert-butyl 2-(2-oxo-2-(pyridin-2-yl)ethyl) pyrrolidine-1,2- dicarboxylate:
  • Step 3 Synthesis of tert-butyl (S)-2-(5-(pyridin-2-yl)-lH-imidazol-2-yl)pyrrolidine-l- carboxylate:
  • step 2 To a stirred solution of (S)-l -tert-butyl 2-(2-oxo-2-(pyridin-2-yl)ethyl) pyrrolidine- 1,2-dicarboxylate (step 2, 13 g, 38.9 mmol) in toluene (130 ml) was added ammonium acetate (29.9 g, 388.3 mmol) at room temperature and refluxed for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated and the crude was diluted with EtOAc. The organic layer was washed with water, brine and then dried over Na 2 S0 4 .
  • Step 2 Synthesis of 1 -(tert-butyl) 2-(2-(3-fluorophenyl)-2-oxoethyl) (S)-pyrrolidine-l,2- dicarboxylate:
  • Step 3 Synthesis of tert-butyl (S)-2-(5-(3-fluorophenyl)-lH-imidazol-2-yl)pyrrolidine-l- carboxylate:
  • Step 4 Synthesis of (S)-5-(3-fluorophenyl)-2-(pyrrolidin-2-yl)-lH-imidazole:
  • Step 1 Synthesis of 2-bromo-l -(3-fluoro-4-meth lphenyl)ethan-l -one:
  • Step 2 Synthesis of l-(tert-butyl) 2-(2-(3-fluoro-4-methylphenyl)-2-oxoethyl) (S)-pyrrolidine- 1 ,2-dicarboxylate:
  • Step 3 Synthesis of tert-butyl (S)-2-(5-(3-fluoro-4-methylphenyl)-lH-imidazol-2- yl)pyrrolidine-l -carboxylate:
  • Step 4 Synthesis of (S)-5-(3-fluoro-4-methylphenyl)-2-(pyrrolidin-2-yl)-lH-imidazole:
  • Step 1 Synthesis of 1 -(4-(benzyloxy)phenyl)ethan-l -one :
  • Step 2 Synthesis of 1 -(4-(benzyloxy)phenyl)-2-bromoethan-l -one:
  • Step 1 To the stirred solution of l-(4-(benzyloxy)phenyl)ethan-l-one (step 1, 25 g, 110.5 mmol) in 200 ml of MeOH at 0°C was added Bromine (4.5 ml, 28.5 mmol) (dropwise addition), stirred for about 30 minutes and stirred for about 4 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated and the crude product was dissolved in n-hexane and stirred for about 30 minutes. The obtained solid was filtered and washed with n-hexane then dried and proceeded for next step (wt: 16.0 g).
  • Step 3 Synthesis of 2-(2-(4-(benzyloxy)phenyl)-2-oxoethyl) l-(tert-butyl) (S)-pyrrolidine-l ,2- dicarboxylate:
  • Step 4 tert-butyl (S)-2-(5-(4-(benzyloxy)phenyl)-lH-imidazol-2-yl)pyrrolidine-l-carboxylate:
  • Step 5 Synthesis of tert-butyl (S)-2-(5-(4-hydroxyphenyl)-lH-imidazol-2-yl)pyrrolidine-l- carboxylate:
  • Step 6 Synthesis of tert-butyl (S)-2-(5-(4-(2-methoxyethoxy)phenyl)-lH-imidazol-2- yl)pyrrolidine-l -carboxylate:
  • reaction mixture was diluted with ethyl acetate, washed with water, IN HC1, water, brine and dried over Na 2 S0 4 .
  • the solvent was evaporated and purified by silica gel column (100-200 mesh, elution 22% acetone: hexane) to afford the title compound (wt: 1.2 g) as an off white solid.
  • Step 7 Synthesis of (S)-5-(4-(2-methoxyethoxy)phenyl)-2-(pyrrolidin-2-yl)-lH-imidazole:
  • Step 1 Synthesis of 2-bromo-l-(pyridin-3-yl) ethanone:
  • Step 2 Synthesis of (S)-l-tert-butyl 2-(2-oxo-2-(pyridin-3-yl)ethyl) pyrrolidine -1,2- dicarboxylate:
  • Step 3 Synthesis of tert-butyl (S)-2-(5-(pyridin-3-yl)-lH-imidazol-2-yl)pyrrolidine-l- carboxylate:
  • step 2 To a stirred solution of (S)-l -tert-butyl 2-(2-oxo-2-(pyridin-3-yl)ethyl) pyrrolidine- 1,2-dicarboxylate (step 2, 13 g, 38.9 mmol) in toulene (130 ml) was added ammonium acetate (29.9 g, 388.3 mmol) at room temperature and refluxed for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated and the crude was diluted with EtOAc. The organic layer was washed with water, brine and then dried over Na 2 S0 4 .
  • Step 1 Synthesis of2-bromo-l-(4-(trifluoromethyl)phenyl)ethan-l-one:
  • Step 2 Synthesis of 1 -(tert-butyl) 2-(2-oxo-2-(4-(trifluoromethyl) phenyl)ethyl) (S)- pyrrolidine-1 ,2-dicarboxylate: To a stirred solution of (S)-l-(tert-butoxycarbonyl)pyrrolidine-2-carboxylic acid (0.4 g, 1.86 mmol) in DCM (20 ml) was added DIPEA (0.64 ml, 4.96 mmol) at 0°C.
  • step 1 2-bromo-l-(4-(trifluoromethyl)phenyl)ethan-l-one (step 1, 0.5 g, 1.86 mmol) was added and the reaction mass was 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 followed by saturated brine and the organic layer was concentrated under reduced pressure. The resulting crude was proceeded to next step without further purification (wt: 0.6 g).
  • Step 3 Synthesis of tert-butyl (S)-2-(5-(4-(trifluoromethyl)phenyl)-lH-imidazol-2- yl)pyrrolidine-l -carboxylate :
  • Step 4 Synthesis of (S)-2-(pyrrolidin-2-yl)-5-(4-(trifluoromethyl)phenyl)-lH-imidazole:
  • Step 1 Synthesis of 2-bromo-l - ⁇ 4 -fluorophenyl) ethan-l-one:
  • Step 2 Synthesis of l-(tert-butyl) 2-(2-(4-fluorophenyl)-2-oxoethyl) (S)-pyrrolidine-l,2- dicarboxylate:
  • Step 3 tert-butyl (S)-2-(5-(4-fluorophenyl)-lH-imidazol-2-yl)pyrrolidine-l-carboxylate:
  • Step 4 Synthesis of (S)-5-(4-fluorophenyl)-2-(pyrrolidin-2-yl)-lH-imidazole:
  • Step 2 Synthesis of (S,Z)-tert-butyl 2-((amino(phenyl)methyleneaminooxy)carbonyl) pyrrolidine-1 -carboxylate: To a stirred solution of Boc-proline (15.8 g, 73.05 mmol) in DCM (200 ml), EDCI (21.05 g, 109.5 mmol) and HOBt (13.4 g, 88.1 mmol) were added at room temperature. After 10 minutes, (Z)-N'-hydroxybenzimidamide (step 1, 10.0 g, 73.05 mmol) was added and the reaction mass was 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 with brine and the organic layer was concentrated under reduced pressure. The resulting crude was proceeded to next step without further purification (wt:
  • Step 3 Synth (S)-tert-butyl 2-(3-phenyl-l ,2,4-oxadiazol-5-yl)pyrrolidine-l - carboxylate:
  • reaction mixture was poured into ice cold water, the solution pH was adjusted to 2 to 3 by addition of 4N HC1 ⁇ Note: the temperature of reaction mixture should be 5-10°C).
  • the aqueous layer was extracted with ethyl acetate (2x500 ml), combined organic layer was washed with brine, dried over Na 2 S0 4i filtered and solvent was evaporated under reduced pressure to provide title compound (wt: 85.0 g, yield: 96%).
  • Step 2 Synthesis of (2S,4R)-l-(tert-butoxycarbonyl)-4-(2-methoxyethoxy)pyrrolidine-2- carboxylic acid:
  • step 1 A solution of (2S,4R)-l-(tert-butoxycarbonyl)-4-hydroxypyrrolidine-2-carboxylic acid (step 1, 20.0 g, 86.9 mmol) in DMF (200 ml) was treated with sodium hydride (8.43 g, 347.8 mmol) at 0°C. After stirring about 15 minutes at 0°C, l-bromo-2-methoxyethane (24.2 g, 174.0 mmol) was added and stirred at ambient temperature for about 18 hours. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0°C, then 400 ml water was added slowly over 30 minutes and stirred at room temperature for about 4 hours.
  • Step 3 Synthesis of (2S,4R)-l-tert-butyl 2-(2-oxo-2-phenylethyl) 4-(2-methoxyethoxy) pyrrolidine-1 ,2-dicarboxylate : To a stirred solution of (2S,4R)-l-(tert-butoxycarbonyl)-4-(2-methoxyethoxy) pyrrolidine -2-carboxylic acid (step 2, 3.20 g, 11.07 mmol) in DCM (50 ml) at 0°C was added DIPEA (3.80 ml, 22.14 mmol).
  • Step 4 Synthesis of (2S,4R)-tert-butyl 4-(2-methoxyethoxy)-2-(4-phenyl-lH-imidazol-2-yl) pyrrolidine-1 -carboxylate:
  • Step 5 Synthesis of2-((2S,4R)-4-(2-methoxyethoxy)pyrrolidin-2-yl)-4-phenyl-lH-imidazole:
  • step 4 To a solution of (2S,4R)-tert-butyl 4-(2-methoxyethoxy)-2-(4-phenyl-lH-imidazol-2- yl) pyrrolidine-1 -carboxylate (step 4, 4.0 g, 10.36 mmol) in DCM (40 ml) was added trifluoroacetic acid (10 ml) and stirred at room temperature for about 4 hours. After completion of the reaction, solvents were evaporated and aqueous sodium bicarbonate was added to the residue. The product was extracted with dichloromethane (2x100 ml), washed with brine, dried over Na 2 S0 4 and filtered.
  • Step 1 Synthesis of (2S,4R)-l-tert-butyl 2-(2-(3-fluorophenyl)-2-oxoethyl) 4-(2-methoxy ethoxy)pyrrolidine-l,2-dicarboxylate:
  • Step 2 Synthesis of (2S,4R)-tert-butyl 2-(4-(3-fluorophenyl)-lH-imidazol-2-yl)-4-(2-methoxy ethoxy)pyrrolidine -1-carboxylate:
  • step 1 To a stirred solution of (2S,4R)-l-tert-butyl 2-(2-(3-fluorophenyl)-2-oxoethyl) 4-(2- methoxyethoxy)pyrrolidine-l,2-dicarboxylate (step 1, 5.0 g, 11.76 mmol) in toluene (50 ml), ammonium acetate (18.11 g, 235.29 mmol) was added at room temperature and resulting mixture was refluxed for about 18 hours. The reaction mixture was cooled to room temperature, added water and extracted with ethyl acetate (2x250 ml) and the organic layer was washed with IN HC1 (2x100 ml).
  • Step 3 Synthesis of 4-(3-fluorophenyl)-2-((2S,4R)-4-(2-methoxyethoxy)pyrrolidin-2-yl)-lH- imidazole 2, 2, 2-trifluoroacetate:
  • step 2 To a solution of ((2S,4R)-tert-butyl 2-(4-(3-fluorophenyl)-lH-imidazol-2-yl)-4-(2- methoxyethoxy)pyrrolidine-l-carboxylate (step 2, 2.50 g, 6.17 mmol) in DCM (30 ml) was added trifluoroacetic acid (12 ml) and stirred at room temperature for about 3 hours. After completion of the reaction, the solvents were evaporated under reduced pressure to afford title compound (wt: 2.58 g, yield: 100%).
  • Step 1 Synthesis of (2S,4R)-l-tert-butyl 2-(2-(4-fluorophenyl)-2-oxoethyl) 4-(2-methoxy ethoxy)pyrrolidine-l,2-dicarboxylate:
  • reaction mixture was diluted with water, extracted with dichloromethane (200 ml) and washed with brine solution (100 ml), dried over Na 2 S0 4 , filtered and concentrated under reduced pressure.
  • the residue was purified on silica gel column (100-200 mesh, eluted with 30% ethyl acetate/hexanes) to afford the title compound (4.5 g, 96%).
  • Step 2 Synthesis of (2S,4R)-tert-butyl 2-(4-(4-fluorophenyl)-lH-imidazol-2-yl)-4-(2-methoxy ethoxy )pyrrolidine-l -carboxylate:
  • step 1 To a stirred solution of (2S,4R)-l-tert-butyl 2-(2-(4-fluorophenyl)-2-oxoethyl) 4-(2- methoxyethoxy)pyrrolidine-l,2-dicarboxylate (step 1, 4.5 g, 10.58 mmol) in toluene (60 ml), ammonium acetate (8.15 g, 105.8 mmol) was added at room temperature and refluxed for about 18 hours. The reaction mixture was cooled to room temperature, diluted with water, extracted with ethyl acetate (200 ml) and the organic layer was washed with IN HCl (2x200 ml).
  • Step 3 Synthesis of 4-(4-fluorophenyl)-2-((2S,4R)-4-(2-methoxyethoxy)pyrrolidin-2-yl)-lH- imidazole 2, 2, 2-trifluoroacetate:
  • Step 1 Synthesis of (2S,4R)-l-tert-butyl 2-(2-oxo-2-(pyridin-3-yl)ethyl) 4-(2-methoxyethoxy) pyrrolidine-1 ,2-dicarboxylate :
  • Step 2 Synthesis of (2S,4R)-tert-butyl 4-(2-methoxyethoxy)-2-(4-(pyridin-3-yl)-lH-imidazol- 2-yl)pyrrolidine-l -carboxylate:
  • step 1 To a stirred solution of (2S,4R)-l-tert-butyl 2-(2-oxo-2-(pyridin-3-yl)ethyl) 4-(2- methoxyethoxy)pyrrolidine-l,2-dicarboxylate (step 1, 16.0 g, 39.21 mmol) in toluene (100 ml), ammonium acetate (24.15 g, 313.72 mmol) was added at room temperature and refluxed for about 24 hours. Reaction mixture was cooled to room temperature, diluted with water, extracted with ethyl acetate (200 ml) and the organic layer was washed with IN HCl (2x150 ml).
  • Step 3 Synthesis of 3-(2-((2S,4R)-4-(2-methoxyethoxy)pyrrolidin-2-yl)-lH-imidazol-4- yl )pyridine 2, 2, 2-trifluoroacetate:
  • step 1 To a mixture of (S)-benzyl 2-(hydrazinecarbonyl)pyrrolidine-l-carboxylate (step 1, 2.20 g, 8.36 mmol), benzoic acid (1.50 g, 12.29 mmol) and HATU ( 4.7 g, 12.36 mmol) in dichloromethane (40 ml) was added diisopropylethyl amine (2.1 g, 32.55 mmol) and stirred at room temperature for about 3 hours. The mixture was diluted with dichloromethane (100 ml), successively washed with water and brine, dried over sodium sulfate and concentrated.
  • Step 3 Synthesis of (S)-Benzyl 2-(5-phenyl-l ,3 ,4-oxadiazol-2-yl)pyrrolidine-l -carboxylate :
  • Step 1 Synthesis of (S)-l-tert-butyl 2-(3,3-dimethyl-2-oxobutyl) pyrrolidine -1,2- dicarboxylate:
  • Step 2 Synthesis of (S)-tert-butyl 2-(4-tert-butyloxazol-2-yl)pyrrolidine-l-carboxylate:
  • step 1 To a stirred solution of (SJ-l-tert-butyl 2-(3,3-dimethyl-2-oxobutyl) pyrrolidine- 1,2- dicarboxylate (step 1, 2.50 g, 7.98 mmol) in toluene (30 ml) was added ammonium acetate (5.53 g, 71.88 mmol) at room temperature and refluxed for about 18 hours. Cooled to room temperature, diluted with ethyl acetate (150 ml) and the organic layer was washed with water, brine (100 ml), dried over Na 2 S0 4 , filtered and the organic layer was concentrated under reduced pressure to afford the title compound (1.2 g, 44% over 2 steps).
  • Step 3 Synthesis of (S)-4-tert-butyl-2-(pyrrolidin-2-yl)oxazole :
  • step 2 1.20 g, 4.09 mmol
  • dichloromethane (10 ml) was added trifluoroacetic acid (2 ml) and stirred at room temperature for about 4 hours. After completion of the reaction, diluted with water and washed with ethyl acetate (2x75 ml). The aqueous layer was basified with aq.
  • Step 2 Synthesis of (S)-tert-butyl 2-(5-methyl-4-phenyl-lH-imidazol-2-yl)pyrrolidine-l- carboxylate:
  • step 2 To a solution of (S)-tert-butyl 2-(l-methyl-4-phenyl-lH-imidazol-2-yl)pyrrolidine-l- carboxylate (step 2, 1.20 g, 3.66 mmol) in dichloromethane (20 ml) was added trifluoroacetic acid (5 ml) and stirred at room temperature for about 4 hours. Reaction mixture was concentrated, aqueous sodium bicarbonate solution was added to the residue and extracted with dichloromethane (2x100 ml), washed with brine, dried over Na 2 S0 4 and filtered. The solvents were evaporated under reduced pressure to afford title compound (660 mg, 79%).
  • Step 1 Synthesis of (S)-tert-butyl 2-(3-methyl-2-oxobutylcarbamoyl)pyrrolidine-l- carboxylate:
  • reaction mixture was extracted with ethyl acetate (2x250 ml) and washed with water (200 ml), saturated brine (100 ml) and the organic layer was concentrated under reduced pressure. The residue was purified by column chromatography (silica gel 100-200 mesh, 30-40% ethyl acetate in hexanes) to afford the title compound (8.10 g, 58%).
  • Step 2 Synthesis of (S)-tert-butyl 2-(4-isopropyl-lH-imidazol-2-yl)pyrrolidine-l-carboxylate:
  • step 1 To a stirred solution of (S)-tert-butyl 2-(3-methyl-2-oxobutylcarbamoyl) pyrrolidine - 1-carboxylate (step 1, 6.0 g, 20.13 mmol) in xylene (100 ml), ammonium acetate (17.0 g, 221.0 mmol) was added at room temperature and heated at 140°C for about 48 hours. The reaction mixture was cooled to room temperature, the pH was adjusted (pH ⁇ 7-8) with aq. NaHC0 3 solution and extracted with ethyl acetate (2x300 ml).
  • step 2 To a solution of (S)-tert-butyl 2-(4-isopropyl-lH-imidazol-2-yl)pyrrolidine-l- carboxylate (step 2, 1.0 g, 3.58 mmol) in dichloromethane (20 ml) was added trifluoroacetic acid (5 ml) and stirred at room temperature for about 4 hours. After completion of the reaction, the solvents were evaporated under reduced pressure to afford title compound (798 mg, 76%).
  • Step 1 Synthesis of (S)-benzyl 2-(2-ox -2-phenylethylcarbamoyl)pyrrolidine-l -carboxylate:
  • Step 2 Synthesis of (S)-benzyl 2-(5-phenyloxazol-2-yl)pyrrolidine-l -carboxylate:
  • step 1 To a stirred solution of (S)-benzyl 2-(2-oxo-2-phenylethylcarbamoyl)pyrrolidine-l- carboxylate (step 1, 1.0 g, 2.73 mmol) in acetonitrile (10 ml) at 0°C was added POCl 3 (2.5 ml, 27.32 mmol) and the resulting solution was heated at 90°C for about 3 hours. The reaction mixture was cooled to room temperature, quenched with ice water and extracted with ethyl acetate (2x50 ml). The organic layer was washed with aq. NaHC0 3 solution, brine, dried over Na 2 S0 4 , filtered and concentrated.
  • Step 1 Synthesis of (S)-benzyl 2-(3,3-dimethyl-2-oxobutylcarbamoyl)pyrrolidine-l- carboxylate:
  • Step 2 Synthesis of (S)-benzyl 2-(5-ter -butyloxazol-2-yl)pyrrolidine-l-carboxylate:
  • step 1 To a stirred solution of (S)-benzyl 2-(3,3-dimethyl-2-oxobutylcarbamoyl)pyrrolidine- 1-carboxylate (step 1, 1.6 g, 4.62 mmol) in acetonitrile (30 ml) at 0°C was added POCl 3 (7.0 g, 46.24 mmol) and resulting solution was heated at 90°C for about 3 hours. The reaction mixture was cooled to room temperature, quenched with ice water and extracted with ethyl acetate (2x100 ml). The organic layer was washed with aq. NaHC0 3 solution, brine, dried over Na 2 S0 4 , filtered and concentrated.
  • Step 3 Synthesis of (S)-5-tert-butyl-2-(pyrrolidin-2-yl)oxazole :
  • Step 1 Synthesis of (S)-l -benzyl 2-(2-oxo-2-phenylethyl) pyrrolidine-1 ,2-dicarboxylate:
  • Step 2 Synthesis of (S)-benzyl 2-(4-phenyloxazol-2-yl)pyrrolidine-l -carboxylate :
  • step 1 To a stirred solution of (S)-l-benzyl 2-(2-oxo-2-phenylethyl) pyrrolidine- 1,2- dicarboxylate (step 1, 12.0 g, 32.69 mmol) in acetic acid (150 ml), ammonium acetate (37.7 g, 490.3 mmol) was added at room temperature and heated at 120°C for about 6 hours. Cooled to room temperature, the reaction mixture was concentrated, the residue was dissolved in dichloromethane (500 ml) and the organic layer was washed with aq. NaHC0 3 solution and brine. The organic layer was dried over Na 2 S0 4 , filtered and solvent was evaporated.
  • step 2 To a solution of (S)-benzyl 2-(4-phenyloxazol-2-yl)pyrrolidine-l -carboxylate (step 2, 3.5 g, 10.05 mmol) in EtOAc (40 ml) was added 10% Pd/C (700 mg) and stirred at room temperature under balloon pressure for about 24 hours. The catalyst was filtered through Celite bed and the filtrate was concentrated to afford the title compound (2.12 g, 98%).
  • Step 1 Synthesis of (S)-tert-butyl 2-(3,3-dimethyl-2-oxobutylcarbamoyl)pyrrolidine-l- carboxylate:
  • step 1 To a stirred solution of (S)-tert -butyl 2-(3,3-dimethyl-2-oxobutylcarbamoyl) pyrrolidine- 1-carboxylate (step 1, 11.3 g, 36.33 mmol) in xylene (130 ml), ammonium acetate (56.0 g, 726.6 mmol) was added at room temperature and heated at 140°C for about 48 hours. The reaction mixture was cooled to room temperature, the pH was adjusted (pH ⁇ 7- 8) with aq. NaHC0 3 solution and extracted with ethyl acetate (2x300 ml).
  • Step 3 Synthesis of (S)-4-tert-butyl-2-(pyrrolidin-2-yl)-lH-imidazole 2,2,2 -trifluoroacetate:
  • Step 1 Synthesis of (S)-l-tert-butyl 2- 2-oxo-2-phenylethyl) pyrrolidine-1 ,2-dicarboxylate:
  • Step 2 Synthesis of (S)-tert-butyl 2-(4-phenyl-lH-imidazol-2-yl)pyrrolidine-l-carboxylate:
  • Step 1 Synthesis of l-(tert-butyl) 2-(2-(4-methoxyphenyl)-2-oxoethyl) (S)-pyrrolidine-l ,2- dicarboxylate:
  • Step 2 Synthesis of tert-butyl (S)-2-(5-(4-methoxyphenyl)-lH-imidazol-2-yl)pyrrolidine-l- carboxylate:
  • step 1 To a stirred solution of 1 -(tert-butyl) 2-(2-(4-methoxyphenyl)-2-oxoethyl) (S)- pyrrolidine-l,2-dicarboxylate (step 1, 7.5 g, 20.71 mmol, 1.0 eq) in toluene (75 mL), ammonium acetate (12.76 g, 165.74 mmol, 8.0 eq) was added at room temperature and refluxed for about 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc and the organic layer was washed with 0.5 N hydrochloric acid.
  • Aqueous layer was basified with 2N NaOH (pH 10-11) and extracted with ethyl acetate. The organic layer was washed with water, dried over Na 2 S0 4 , filtered and solvent was evaporated under reduced pressure to provide a light yellow solid was taken in to hexane, stirred for one hour and filtered (4.0 g, yield: 59.0%).
  • Step 1 Synthesis of 1 -(tert-butyl) 2-(2-(4-chlorophenyl)-2-oxoethyl) (S)-pyrrolidine-l,2- dicarboxylate:
  • Step 2 Synthesis of tert-butyl (S)-2-(5-(4-chlorophenyl)-lH-imidazol-2-yl)pyrrolidine-l- carboxylate:
  • step 1 8.0 g, 21.85 mmol
  • ammonium acetate (13.4 g, 174.8 mmol) was added at room temperature and refluxed for about 16 hours.
  • the reaction mixture was diluted with EtOAc and the organic layer was washed with 0.5 N hydrochloric acid.
  • Aqueous layer was basified with 2N NaOH (pH 10-11) and extracted with ethyl acetate. The organic layer was washed with water, dried over Na 2 S0 4 , filtered and the solvent was evaporated under reduced pressure to afford the title compound (5.0 g, yield: 67.0%).
  • Step 1 Synthesis of (S)-l-(tert-butoxycarbonyl)piperidine-3-carboxylic acid: To a stirred solution of (S)-(+)-3-piperidine carboxylic acid (5.0 g, 38.75 mmol, 1.0 eq) in 1,4-dioxane (50 ml) was added IN NaOH (25 mL) and (Boc) 2 0 (12.6 gr, 58.1 mmol). The reaction mixture was stirred at room temperature for overnight. TLC indicated starting material was consumed and the desired product was observed.
  • Step 2 Synthesis of l-(tert-butyl) 3-(2-oxo-2-phenylethyl) (S)-piperidine-l, 3 '-dicarboxylate:
  • step 1 To a stirred solution of (S)-l-(tert-butoxycarbonyl)piperidine-3-carboxylic acid (step 1, 5.3 g, 23.1 mmol, 1.0 eq) in CH 2 C1 2 (53 ml) was added DIPEA (11.93 gr, 92.45 mmol) and 2-bromoacetophenone (13.02 gr, 6.07 mmol). The reaction mixture was stirred at room temperature for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was diluted with water. The aqueous layer was extracted with CH 2 C1 2 (2x100 ml).
  • Step 3 Synthesis of tert-butyl (S)-3-(5-phenyl-lH-imidazol-2-yl)piperidine-l-carboxylate:
  • Step 4 Synthesis of (S)-3-(5-phenyl-lH-imidazol-2-yl)piperidin-l-ium chloride:
  • Step 1 Synthesis of 1 -(tert-butyl) 4-ethyl piperidine-1 ,4-dicarboxylate :
  • Step 2 Synthesis of 1 -(tert-butoxycarbonyl)piperidine-4-carboxylic acid:
  • Step 3 Synthesis of l-(tert-butyl) 4-(2-oxo-2-phenylethyl) piperidine-l ,4-dicarboxylate:
  • Step 4 Synthesis of tert-butyl 4-(5-phenyl- -imidazol-2-yl)piperidine- 1 -carboxylate :
  • Step 5 Synthesis of 4-(5 -phenyl- lH-imidazol-2-yl)piperidine hydrochloride:
  • step 4 To a stirred solution of tert-butyl 4-(5-phenyl-lH-imidazol-2-yl)piperidine-l- carboxylate (step 4, 2.2 g, 6.72 mmol, 1.0 eq) in CH 2 C1 2 (22 ml) was added 4M HCl in dioxane (20 mL). The reaction mixture was stirred at room temperature for 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 (1.52 gr) and used directly for next step without further purification.
  • Step 3 Synthesis ofN-(l-benzylpiperidin-4- l)isobutyramide:
  • step 2 To a stirred solution of l-benzylpiperidin-4-amine (step 2, 2.9 g, 15.26 mmol, 1.0 eq) in CH 2 C1 2 (40 ml) at 0°C was added triethylamine (5.46 ml, 39.4 mmol, 3.0 eq) and isobutyryl chloride (1.62 g, 15.2 mmol, 1.0 eq). The reaction mixture was allowed to stir at room temperature for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was washed with water (2x50 ml), dried over Na 2 S0 4 , filtered and evaporated under reduced pressure.
  • Step 5 Synthesis of 1 -benzyl-4-(3-isopropyl-5 -methyl-4H-l ,2,4-triazol-4-yl)piperidine :
  • step 1 To a solution of L-Proline methyl ester (step 1, 1 mol) in CH 2 CI 2 was added B0C 2 O (1.5mol), Et 3 N (3mmol), and DMAP (1 mol) at 0°C under Ar. After 18 hours, the crude mixture was washed with aqueous HCl 0.5 M, saturated aqueous NaHC0 3 solution, 3 ⁇ 40, and brine. The organic phase was dried over Na 2 S0 4 , filtered, concentrated in vacuo, and the residue was recrystallized using ether to afford compound.
  • step 2 (2S)-N-(tert-Butoxycarbonyl) proline methyl ester (step 2) was heated in a mixture of THF and 28% aqueous NH 3 (2/9, v/v) at 60°C for about 18 hours under Ar. The reaction mixture was concentrated to give Boc-L-Pro-N3 ⁇ 4, used in the next step without any further purification
  • Step 5 Synthesis of tert-butyl-2-(5-phenyl-4H-l, 2, 4-triazole)pyrrolidine-l-carboxylate: I _ N— N
  • Step 1 Synthesis of (S)-l -tert-butyl 2-(2-(3-fluoro-4-methoxyphenyl)-2-oxoethyl) pyrrolidine- 1 ,2-dicarboxylate:
  • the reaction mixture was diluted with dichloromethane (200 ml), washed with water, saturated brine, dried over Na 2 S0 4 , filtered and the organic layer was concentrated under reduced pressure to afford the title compound (crude 7.5 g, 97%).
  • the crude product was used in the next step without further purification.
  • Step 2 Synthesis of (S)-tert-butyl 2-(4-(3-fluoro-4-methoxyphenyl)-lH-imidazol-2- yl pyrrolidine -1 -carboxylate:
  • step 1 To a stirred solution of (S)-l-tert-butyl 2-(2-(3-fluoro-4-methoxyphenyl)-2-oxoethyl) pyrrolidine- 1,2-dicarboxylate (step 1, 7.5 g, 19.68 mmol) in toluene (60 ml), ammonium acetate (15.15 g, 196.8 mmol) was added at room temperature and refluxed for about 18 hours. Cooled to room temperature, the reaction mixture was diluted with ethyl acetate (200 ml) and the organic layer was washed with 1 N HC1 (2x100 ml).
  • Step 3 Synthesis of (S)-4-(3-fluoro-4-methoxyphenyl)-2-(pyrrolidin-2-yl)-lH-imidazole:
  • Step 1 Synthesis of tert-butyl (S)-2-(hydroxymethyl)pyrrolidine-l -carboxylate :
  • Step 2 Synthesis of tert-butyl ( S)-2-formylpy rrolidine-1 -carboxylate :
  • step 2 To a solution of tert-butyl (S)-2-formylpyrrolidine-l-carboxylate (step 2, 1.6 g, 8.03 mmol, 1.0 eq) in MeOH (20 ml) was added 3,3-dibromo-l,l,l-trifluoropropan-2-one (3.4 ml, 99.03 mmol, 3.7 eq) and aq. NH 3 (1.1 ml 4.16 mmol, 1.09 eq) in one portion. The reaction mixture was stirring at room temperature for about 18 hours.
  • Step 4 Synthesis of 2,2,2-trifluoro-l-((2S)-2-(5-(trifluoromethyl)-lH-imidazol-2-yl)-lX 4 - pyrrolidin-1 -yl)ethan-l -one:
  • step 3 (trifluoromethyl)-lH-imidazol-2-yl)pyrrolidine-l-carboxylate (step 3, 1.9 g, 6.2 mmol, 1) in DCM (30 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the solvent was evaporated under reduced pressure to give the title compound, which was directly used in the next step.
  • step 1 To a solution of (S)-tert-butyl 2-(lH-benzo[d]imidazol-2-yl)pyrrolidine-l-carboxylate (step 1, 2.0 g, 6.90 mmol) in DCM (20 ml) was added trifluoroacetic acid (2.6 ml, 34.84 mmol) and stirred at room temperature for about 2 hours. After completion of the reaction (monitored by TLC), the solvents were evaporated under reduced pressure to afford title compound (2.0 g, yield: 100%) as a TFA salt.
  • Step 1 Synthesis of 2-bromo-l-(2,4-difluorophenyl)ethan-l -one:
  • Step 2 Synthesis of l-(tert-butyl) 2-(2-(2,4-difluorophenyl)-2-oxoethyl) (S)-pyrrolidine-l,2- dicarboxylate:
  • reaction mixture was diluted with DCM, washed with water, saturated brine and the organic layer was concentrated under reduced pressure, the resulting crude was proceeded to next step without further purification (wt: 18.0 g).
  • Step 3 Synthesis of tert-butyl (S)-2-(5-(2,4-difluorophenyl)-lH-imidazol-2-yl)pyrrolidine-l- carboxylate:
  • Step 4 Synthesis of (S)-5-(2,4-difluorophenyl)-2-(pyrrolidin-2-yl)-lH-imidazole:
  • Step 1 Synthesis of 2-bromo-l -(2-fluorophenyl)ethan-l -one:
  • Step 2 Synthesis of l-(tert-butyl) 2-(2-(2-fluorophenyl)-2-oxoethyl) (S)-pyrrolidine-l,2- dicarboxylate:
  • reaction mixture was diluted with DCM, washed with water, saturated brine and the organic layer was concentrated under reduced pressure, the resulting crude was proceeded to next step without further purification (wt: 18.0 g).
  • Step 3 Synthesis of tert-butyl (S)-2-(5-(2-fluorophenyl)-lH-imidazol-2-yl)pyrrolidine-l- carboxylate:
  • Step 4 Synthesis of (S)-5-(2-fluorophenyl)-2-(pyrrolidin-2-yl)-lH-imidazole: A solution of tert-butyl (5)-2-(5-(2-fluorophenyl)-lH-imidazol-2-yl)pyrrolidine-l- carboxylate (step 3, 2.1 g, 6.4 mmol) in TFA:DCM (1:2, 30 mL) stirred at 0 °C - room temperature for about 1 hour. After completion of the reaction, the solvent was evaporated and the crude was dissolved in DCM and proceeded for next step without further purification (wt: 1.4 g).
  • Step 1 Synthesis of (S)-l -tert-butyl 2-(2-(3-chlorophenyl)-2-oxoethyl) pyrrolidine-1,2- dicarboxylate:
  • Step 2 Synthesis of (S)-tert-butyl 2-(4-(3-chlorophenyl)-lH-imidazol-2-yl)pyrrolidine-l- carboxylate:
  • step 1 To a stirred solution of (S)-l-tert-butyl 2-(2-(3-chlorophenyl)-2-oxoethyl) pyrrolidine - 1,2-dicarboxylate (step 1, 16.0 g, 43.71 mmol) in toluene (150 ml), ammonium acetate (33.70 g, 437.15 mmol) was added at room temperature and resulting reaction mixture was refluxed for about 18 hours. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to room temperature, water was added, extracted with ethyl acetate (200 ml) and the organic layer was washed with IN HC1 (2x100 ml).
  • step 2 To a solution of (S)-tert-butyl 2-(4-(3-chlorophenyl)-lH-imidazol-2-yl)pyrrolidine-l- carboxylate (step 2, 2.0 g, 5.78 mmol) in DCM (20 ml) was added trifluoroacetic acid (5.7 ml, 74.51 mmol) and stirred at room temperature for about 18 hours. After completion of the reaction (monitored by TLC), the solvents were evaporated under reduced pressure to afford the title compound (2.0 g) as a TFA salt.
  • Step 1 Synthesis of (2S,4R)-1 -tert-butyl 2-(2-oxo-2-phenylethyl) 4-hydroxypyrrolidine-l ,2- dicarboxylate:
  • Step 2 Synthesis of (2S,4R)-tert-butyl 4-hydroxy-2-(4-phenyl-lH-imidazol-2-yl)pyrrolidine- 1-carboxylate:
  • step 1 stirred solution of (2S,4R)-l-tert-butyl 2-(2-oxo-2-phenylethyl) 4-hydroxy pyrrolidine- 1,2-dicarboxylate (step 1, 20.0 g, 57.5 mmol) in toluene (200 ml), ammonium acetate (44.25 g, 575.0 mmol) was added at room temperature and resulting reaction mixture was refluxed for about 18 hours. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to room temperature, water was added and extracted with ethyl acetate (2x250 ml) and the organic layer was washed with IN HC1 (2x200 ml).
  • Step 2 Synthesis of (2S,4R)-1 -tert-butyl 2-(2-oxo-2-phenylethyl) 4-isopropoxypyrrolidine- 1 ,2-dicarboxylate:
  • step 1 To a stirred solution (2S,4R)-l-(tert-butoxycarbonyl)-4-isopropoxypyrrolidine-2- carboxylic acid (step 1, 10.0 g, 36.6 mmol) in DCM (100 ml) at 0 °C was added triethyl amine (11.1 g, 109.9 mmol) and after stirring about 10 minutes, 2-bromo-l-phenylethanone (8.74 g, 43.9 mmol) was added and the reaction mixture was allowed to stir at room temperature for about 18 hours. After completion of the reaction (monitored by TLC), water was added to the reaction mixture and extracted with dichloromethane (200 ml).
  • Step 4 Synthesis of 2-((2S,4R)-4-isopropoxypyrrolidin-2-yl)-4-phenyl-lH-imidazole TFA salt:
  • Step 1 Synthesis of (2S,4R)-tert-butyl 2-(3,3-dimethyl-2-oxobutylcarbamoyl)-4-(2-methoxy ethoxy )pyrrolidine-l -carboxylate:
  • Step 2 Synthesis of (2S,4R)-tert-butyl 2-(4-tert-butyl-lH-imidazol-2-yl)-4-(2-methoxyethoxy) pyrrolidine-1 -carboxylate:
  • Step 3 Synthesis of 4-tert-butyl-2-((2S,4R)-4-(2-methoxyethoxy)pyrrolidin-2-yl)-lH- imidazole TFA salt: To a solution of (2S,4R)-tert-butyl 2-(4-tert-butyl-lH-imidazol-2-yl)-4-(2-methoxy ethoxy)pyrrolidine-l-carboxylate (step 2, 3.0 g, 8.19 mmol) in DCM (30 ml) was added trifluoroacetic acid (15 ml) and stirred at room temperature for about 3 hours.
  • Step 2 Synthesis of (2S,4R)-1 -tert-butyl 2-(2-oxo-2-phenylethyl) 4-methoxypyrrolidine-l ,2- dicarboxylate:
  • reaction mixture was diluted with dichloromethane (200 ml), washed with water (200 ml), saturated brine (100 ml), dried over Na 2 S0 4 , filtered and concentrated under reduced pressure. The residue was purified on silica gel column (100-200 mesh, eluted in 20-25% ethyl acetate/hexane) to afford the title compound (6.80 g, yield: 77%).
  • Step 3 Synthesis of (2S,4R)-tert-butyl 4-methoxy-2-(4-phenyl-lH-imidazol-2-yl)pyrrolidine- 1-carboxylate:
  • Step 1 Synthesis of (2S,4R)-l-tert-butyl 2-(2-(3-fluorophenyl)-2-oxoethyl) 4-methoxy pyrrolidine-1 ,2-dicarboxylate :
  • reaction mixture was diluted with dichloromethane (200 ml) and washed with water (200 ml), saturated brine (100 ml), dried over Na 2 S0 4 , filtered and concentrated under reduced pressure. The residue was purified on silica gel column (100-200 mesh, eluted in 25% ethyl acetate/hexanes) to afford the title compound (1.8 g, yield: 30%).
  • Step 2 Synthesis of (2S,4R)-tert-butyl 2-(4-(3 -fluorophenyl)-! H-imidazol-2-yl)-4-methoxy pyrrolidine-! -carboxylate:
  • Step 1 Synthesis of (S)-tert-butyl 2-(4,5-dimethyl-lH-imidazol-2-yl)pyrrolidine-l- carboxylate:
  • step 1 To a solution of (S)-tert-butyl 2-(4,5-dimethyl-lH-imidazol-2-yl)pyrrolidine-l- carboxylate (step 1, 3.70 g, 14.01 mmol) in DCM (40 ml) was added trifluoroacetic acid (20 ml) and stirred at room temperature for about 5 hours. After completion of the reaction (monitored by TLC), the solvents were evaporated under reduced pressure to afford title compound (3.80 g) as a TFA salt.
  • Step 1 Synthesis of (S)-tert-butyl 2-(lH-imida ol-2-yl)pyrrolidine-l -carboxylate :
  • 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-l-(prop-l-en-2-yl)-3a-((S)-2-(5-(pyridin-2-yl)-lH- imidazol-2-yl)pyrrolidine-l-carbonyl)icosahvdro-lH-cvclopentaralchrvsen-9-yl)oxy) carbonvDcyclobutane-l-carboxylic acid:
  • Step 1 Synthesis of (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla-penta methyl-l-(prop-l-en-2-yl)-3a-((S)-2-(5-(pyridin-2-yl)-lH-imidazol-2-yl)pyrrolidine-l- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate:
  • reaction mixture was diluted with DCM, washed with water followed by IN HC1, water, brine and dried over Na 2 S0 4 .
  • the solvent was evaporated and purified by silica gel column (100-200 mesh, elution 5%MeOH/DCM) to afford the title compound (2.8 g) as an off white solid.
  • Step 2 Synthesis of ((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-9-hydroxy-5a,5b,8,8, lla-pentamethyl-l-(prop-l-en-2-yl)icosahydro-3aH-cyclopenta[a]chrysen-3a-yl)((S)-2-(5- (pyridin-2-yl)-lH-imidazol-2-yl)pyrrolidin-l-yl)methanone:
  • Step 3 Synthesis of 1 -benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8, lla ⁇ entamethyl-l-(prop-l-en-2-yl)-3a-((S)-2-(5-(pyridin-2-yl)-lH-imidazol-2-yl)pyrrolidine -l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-dimethylcyclobutane-l,3 -dicarboxylate:
  • the reaction mixture was heated FOR overnight at 90°C. 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 Cl 2 (2x50 ml), the DCM layer was washed with IN HC1, followed by water and brine solution.
  • the 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 1.5% methanol: dichloromethane as an eluent to obtain the title compound (0.5 g) as a white solid.
  • 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-l-(prop-l-en-2-yl)-3a-((S)-2-(5-(pyridin-2-yl)-lH-imidazol -2-yl)pyrrolidine-l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclo butane-1 -carboxylic acid:
  • Example 3 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-(prop-l-en-2-yl)-3a-((S)-2-(5-(pyridin-3-yl)-lH- imidazol-2-yl)pyrrolidine-l-carbonyl)icosa ydro-lH-cyclopentaralchrysen-9-yl)oxy) carbonyl)cyclobutane-l-carboxylic acid:
  • Example 7 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a -((2S ⁇ R)-4-(2-methoxyethoxy)-2-(4-phenyl-lH-imidazol-2-yl)pyrrolidine-l-carbonyl)-5a, 5b, 8, 8 J la-pentamethyl-l-(prop-l-en-2-yl)icosa vdro-lH-cvclopentaralchrvsen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane- 1-carboxylic acid:
  • Example 8 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a -((2S,4R)-2-(4-(3-fluorophenyl)-lH-imidazol-2-yl)-4-(2-methoxyethoxy)pyrrolidine-l- carbonyl)-5a,5b,8,8 J la-pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen -9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l-carboxylic acid:
  • Example 10 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((2S,4R)-4-(2-methoxyethoxy)-2-(4-(pyridin-3-yl)-lH-imidazol-2-yl)pyrrolidine-l- carbonyl)-5a,5b,8,8 J la-pentamethyl-l-(prop-l-en-2-yl)icosahvdro-lH-cvclopentaralchrvsen - -yl)oxy)carbonyl)-2,2-dimethylcvclobutane-l-carboxylic acid:
  • Example 11 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)-2-(5-phenyl-l,3,4-oxadiazol-2-yl)pyrrolidine -l-carbonyl)-l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)carbonyl) cyclobutane-l-carboxylic acid:
  • Example 12 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((S)-2-(4-(tert-butyl)oxazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b,8,8 J la-pentamethyl-1- (prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclo butane- 1-carboxylic acid:
  • 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-((S)-2-(5-methyl-4-phenyl-lH-imidazol-2-yl) pyrrolidine- l-carbonyl)-l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy) carbonyl)cyclobutane-l-carboxylic acid:
  • Example 14 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((S)-2-(4-isopropyl-lH-imidazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b,8,8 J la-pentamethyl- l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)carbonyl)-2,2-dimethyl cyclobutane-l-carboxylic aci
  • Example 15 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)-2-(5-phenyloxazol-2-yl)pyrrolidine-l- carbonyl)-l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)carbonyl)cyclo butane- 1-carboxylic acid:
  • Example 16 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((S)-2-(5-(tert-butyl)oxazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b,8,8 J la-pentamethyl-1- (prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)carbonyl)-2,2-dimethylcy butane- 1-carboxylic acid:
  • Example 17 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)-2-(4-phenyloxazol-2-yl)pyrrolidine-l- carbonyl)-l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)carbonyl)cyclo butane- 1-carboxylic acid:
  • Example 18 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((S)-2-(4-(tert-butyl)-lH-imidazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b,8,8 J la-penta methyl- l-(prop-l-en-2-yl)icosahvdro-lH-cvclopentaralchrvsen-9-yl)oxy)carbonyl)-2,2- dimethylcyclobutane-l-carboxylic acid:
  • Example 20 Preparation of (lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR, 13aRJ3bR)-5a,5b,8,8 J la-pentamethyl-3a-((S)-3-(5-phenyl-lH-imidazol-2-yl)piperidine-l- carbonyl)-l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)carbonyl)cyclo butane- 1-carboxylic acid:
  • Example 21 Preparation of (lR,3R)-2,2-dimethyl-3-(2-oxo-2-(((lR,3aS,5aR,5bR,7aR,9S, l laR,l lbR,13aR,13bR)-5a,5b,8,8,l la-pentamethyl-3a-((S)-2-(5-phenyl-lH-imidazol-2-yl) pyrrolidine- l-carbonyl)-l-(prop-l-en-2-yl)icosanydro-lH-cyclopentaralchrysen-9-yl)oxy) ethyl)cyclobutane-l-carboxyric acid:
  • Example 22 Preparation of 2.2-dimethyl-3-((((lR.3aS.5aR.5bR.7aR.9S.l laR.l lbR.13aR. 13bR)-5a.5b.8.8.11a-pentamethyl-3a-((S)-2-(5-phenyl-lH-imidazol-2-yl)pyrrolidine-l- carbonyl)-l-(prop-l-en-2-yl)-icosahvdro-lH-cvclopentaralchrvsen-9-yl)oxy)carbonyl)cvclo propane- 1-carboxylic acid:
  • Example 23 Preparation of 2-methyl-2-(4-((((lR.3aS,5aR.5bR.7aR.9S,l laR.l lbR.13aR.
  • Example 24 Preparation of (lR.3S)-2.2-dimethyl-3-((((lR.3aS.5aR.5bRJaR.9S.l laR.l lbR. 13aR.13bR)-5a.5b.8.8.11a-pentamethyl-3a-((S)-2-(5-phenyl-4H-1.2.4-triazol-3-vnpyrrolidine - 1 -carbonyl)- 1 -(prop- 1 -en-2-yl)icosahydro- lH-cvclopentara1chrvsen-9-yl)oxy)carbonyl) cyclobutane-l-carboxylic acid:
  • Example 25 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((S)-2-(4-(3-fluoro-4-methoxyphenyl)-lH-imidazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b,8, 8 J 1 a-pentamethyl- 1 -(prop- 1 -en-2-yl)icosahydro- 1 H-cyclopentaral chry sen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane- -carboxylic acid:
  • Example 26 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((S)-2-(5-(2,4-difluorophenyl)-lH-imidazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la- pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)carbonyl)-2,2- dimethylcyclobutane- 1-carboxylic acid:
  • Example 28 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((S)-2-(4-(3-chlorophenyl)-lH-imidazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b,8,8 J la-penta methyl- l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)carbonyl)-2,2- dimethylcyclobutane-l-carboxylic acid:
  • Example 29 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((2S,4R)-4-hydroxy-2-(4-phenyl-lH-imidazol-2-yl)pyrrolidine-l-carbo
  • Example 30 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((2S,4R)-4-isopropoxy-2-(4-phenyl-lH-imidazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b,8,8, l la-pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)carbonyl)- 2,2-dimethylcyclobutane- 1-carboxylic acid:
  • Example 32 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((2S ⁇ R)-4-methoxy-2-(4-phenyl-lH-imidazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b,8,8J la -pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)carbonyl)- 2,2-dimethylcyclobutane- 1-carboxylic acid:
  • Example 33 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((2S,4R)-2-(4-(3-fluorophenyl)-lH-imidazol-2-yl)-4-methoxypyrrolidine-l-carbonyl)-5a, 5b, 8, 8 J la-pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane- 1-carboxylic acid:
  • Example 34 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-(prop-l-en-2-yl)-3a-((S)-2-(5-(trifluoromethyl)-lH -imidazol-2-yl)pyrrolidine-l-caA
  • Example 36 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((S)-2-(lH-imidazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b,8,8 J la-pentamethyl-l-(prop-l- en-2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid:
  • Example 37 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((S)-2-(lH-benzordlimidazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b,8,8 J la-pentamethyl-1- (prop-l-en-2-yl)icosahvdro-lH-cvclopentaralchrvsen-9-yl)oxy)carbonyl)-2,2-dimethylcvclo
  • Example 38 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((lR,3R,5S)-3-(3-isopropyl-5-methyl-4H-l,2,4-triazol-4-yl)-8-azabicvclor3.2.11octane-8- carbonyl)-5a,5b,8,8 J la-pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen -9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l-carboxylic acid:
  • Example 39 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-l-(prop-l-en-2-yl)-3a-((S)-2-(5-(pyridin-2-yl)-lH- imidazol-2-yl)pyrrolidine-l-carbonyl)icosah ⁇
  • reaction mixture was diluted with EtOAc, washed with water followed by IN HC1, water and brine solution. The residue was dried over Na 2 S0 4 , the solvent was evaporated under vacuum and purified by silica gel column (100-200 mesh, elution 5% MeOH/DCM) to afford the title compound (wt: 0.120 g) as an off white solid.
  • Example 40 Preparation of 4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2- (5-(3-fluoro-4-methylphenyl)-lH-imidazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b,8,8 J la-penta methyl- l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)-2,2-dimethyl-4-oxo butanoic acid:
  • Example 41 Preparation of 5-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2- (5-(4-fluorophenyl)-lH-imidazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b,8,8 J la-pentamethyl-1- (prop-l-en-2-yl)icosa vdro-lH-cvclopentaralchrvsen-9-yl)oxy)-3,3-dimethyl-5-oxopentanoic acid:
  • Example 42 Preparation of 4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bS)-l-(l,2- dihydroxypropan-2-yl)-5a,5b,8,8 J la-pentamethyl-3a-((R)-2-(3-phenyl-l,2,4-oxadiazol-5-yl) pyrrolidine- l-carbonyl)icosa ydro-lH-cyclopentaralchrysen-9-yl)oxy)-2,2-dimethyl-4-oxo butanoic acid:
  • Example 43 Preparation of 4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((2S, 4R)-4-(2-methoxyethoxy)-2-(4-phenyl-lH-im l la-pentamethyl-l-(prop-l-en-2-yl)icosa vdro-lH-cvclopentaralchrvsen-9-yl)oxy)-2,2- dimethyl-4-oxobutanoic acid:
  • Example 44 Preparation of 4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((2S, 4R)-2-(4-(3-fluorophenyl)-lH-imidazol-2-yl)-4-(2-methoxyethoxy)pyrrolidine-l-carbonyl)- 5a,5b,8,8 J la-pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy) -2,2-dimethyl-4-oxobutanoic acid:
  • Example 45 Preparation of 4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((2S, 4R)-2-(4-(4-fluorophenyl)-lH-imidazol-2-yl)-4-(2-methoxyethoxy)pyrrolidine-l-carbonyl)- 5a,5b,8,8 J la-pentamethyl-l-(prop-l-en-2-yl)icosahvdro-lH-cvclopentaralchrvsen-9-yl)oxy) -2,2-dimethyl-4-oxobutanoic acid:
  • Example 46 Preparation of 4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2- (5-(4-methoxyphenyl)-lH-imidazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b,8,8 J la-pentamethyl- l-(prop-l-en-2-yl)icosahvdro-lH-cvclopentaralchrvsen-9-yl)oxy)-2,2-dimethyl-4-oxo butanoic acid:
  • Example 47 Preparation of 2,2-dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR, 13aRJ3bR)-5a,5b,8,8J la-pentamethyl-3a-((S)-3-(5-phenyl-lH-imidazol-2-yl)piperidine-l- carbonyl)-l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)butanoic acid:
  • Example 48 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-(5-phenyl-4H-l,2,4-triazol-3-yl)pyrrolidine -l-carbonyl)-l-(prop-l-en-2-yl)icosahvdro-lH-cvclopentaralchrvsen-9-yl)oxy)butanoic acid:
  • Example 49 Preparation of 4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((lR, 3R,5S)-3-(3-isopropyl-5-methyl-4H-l,2,4-triazol-4-yl)-8-azabicvclor3.2.11octane-8- carbonyl)-5a,5b,8,8 J la-pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen -9-yl)oxy)-2,2-dimethyl-4-oxobutanoic acid:
  • Example 50 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((S)-2-(5-(3-fluorophenyl)-lH-imidazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b,8,8 J la-penta methyl- l-(l-methylcvclopropyl)icosahvdro-lH-cvclopentaralchrvsen-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-((S)-2-(5-(3-fluoro phenyl)-! H-imidazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b, 8,8 ,11 a-pentamethyl-1 -( 1 -methyl cyclopropyl)icosahydro-lH-cyclopenta[a ]chrysen-9-yl acetate:
  • Step 2 Synthesis of ((S)-2-(5-(3-fluorophenyl)-lH-imidazol-2-yl)pyrrolidin-l-yl)((lR,3aS, 5aR, 5bR, 7aR, 9S, 11 aR, 11 bR, 13aR, 13bR )-9-hydroxy-5a, 5b, 8, 8, 1 la-pentamethyl-1 -( 1 -methyl cyclopropyl)icosahydro-3aH-cyclopenta[a]chrysen-3a-yl)methanone:
  • Step 3 Synthesis of 1 -benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-3a-((S)-2- (5-(3 -fluorophenyl)-! H-imidazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b, 8, 8,11 a-pentamethyl-1 - (1 -methylcyclopropyl)icosahydro-lH-cyclopenta[a] chrysen-9-yl) (lR,3S)-2,2-dimethylcyclo butane -1, 3 -dicarboxy late:
  • the reaction mixture was heated for overnight at 90°C. 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 (2x50 ml), the DCM layer was washed with IN HC1, followed by water and brine solution.
  • the 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 1.5% methanol: dichloromethane as an eluent to afford the title compound (wt: 0.3g) as a white solid.
  • Step 4 Synthesis of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-3a-((S)-2- (5-(3 -fluorophenyl)-! H-imidazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b,8,8,lla-pentamethyl-l- (l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl cyclobutane-1 -carboxylic acid:
  • Example 51 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-((S)-2-(5-(pyridin-2-yl)- lH-imidazol-2-yl)pyrrolidine-l-carbonyl)icosahydro-lH-cyclopentaralchrysen-9-y carbonvDcyclobutane-l-carboxylic acid:
  • Example 52 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((S)-2-(5-(4-(2-methoxyethoxy)phenyl)-lH-imidazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b, 8,8 J la-pentamethyl-l-(l-methylcvclopropyl)icosahvdro-lH-cvclopentaralchrvsen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane- 1-carbox lic acid
  • Example 53 Preparation of 3-(2-((S)-l-((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9- (((lS,3R)-3-carboxy-2,2-dimethylcyclobutane-l-carbonyl)oxy)-5a,5b,8,8 J la-pentamethyl-1- (l-methylcyclopropyl)icosahydro-lH-cyclopentaralchrysene-3a-carbonyl)pyrrolidin-2-yl)-lH -imidazol-5-yl)pyridine 1 -oxide:
  • Example 54 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3 a-((2S ,4R)-2-(4-( 3 -fluorophenyl) - 1 H-imidazol-2- yl)-4-( 2-methoxyethoxy)p yrrolidine- 1 - carbonyl)-5a,5b,8,8 J la-pentamethyl-l-(l-methylcyclopropyl)icosahydro-lH-cyclopentaral chrysen-9-yl)oxy)carbonyl)-2, -dimethylcyclobutane-l-carboxylic acid:
  • Example 55 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((2S,4R)-2-(4-(4-fluorophenyl)-lH-imidazol-2-yl)-4-(2-methoxyethoxy)pyrrolidine-l- carbonyl)-5a,5b,8,8 J la-pentamethyl-l-(l-methylcyclopropyl)icosahydro-lH-cyclopentaral chrysen-9-yl)oxy)carbonyl)-2 -dimethylcyclobutane-l-carboxylic acid:
  • Example 56 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((S)-2-(4-(tert-butyl)-lH-imidazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b,8,8 J la-penta methyl- l-(l-methylcyclopropyl)icosa ydro-lH-cyclopentaralchrysen-9-yl)oxy)carbonyl)-2,2 -dimethylcyclobutane- 1-carboxylic acid:
  • Example 57 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-(4-(5-phenyl-lH- imidazol-2-yl)piperidine-l-carbonyl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)carbonyl) eye lobutane- 1-carboxylic acid:
  • Example 58 Preparation of (lR.3S)-3-((((lR.3aS.5aR.5bR.7aR.9S.l laR.l lbR.13aR.13bR)- 3a-(4-(3-isopropyl-5-methyl-4H-L2 ⁇ -triazol-4-yl)piperidine-l-carbonyl)-5a,5b,8,8 J la- pentamethyl-l-(l-methylcyclopropyl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane- 1-carboxylic acid:
  • Example 59 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-((S)-2-(5-(trifluoro methyl)- lH-imidazol-2-yl)pyrrolidine-l-carbonyl)icosa ydro-lH-cyclopentaralchrysen-9-yl) oxy)carbonyl)cyclobutane- 1-carboxylic acid:
  • Example 60 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((lR,3R,5S)-3-(3-isopropyl-5-methyl-4H-l,2,4-triazol-4-yl)-8-azabicvclor3.2.11octane-8- carbonyl)-5a,5b,8,8 J la-pentamethyl-l-(l-methylcyclopropyl)icosahydro-lH-cyclopentaral chrvsen-9-yl)oxy)carbonyl)- -dimethylcyclobutane- 1-carboxylic acid:
  • Example 61 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((S)-2-(lH-benzordlimidazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b,8,8 J la-pentamethyl-1- (l-methylcyclopropyl)icosahydro-lH-cy ⁇
  • Example 62 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-l-(l-methylcvclopropyl)-3a-((S)-2-(5-(pyridin-2-yl)- lH-imidazol-2-yl)pyrrolidine-l-carbonyl)icosahvdro-lH-cvclopentaralchrysen-9-yl)oxy) butanoic acid:
  • Step 1 Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla-penta methyl-l-(l -methylcyclopropyl)-3a-((S)-2-(5-(pyridin-2-yl)-lH-imidazol-2-yl)pyrrolidine-l- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate:
  • Step 2 Synthesis of ((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-9-hydroxy-5a,5b,8,8, lla ⁇ entamethyl-l-(l-methylcyclopropyl)icosahydro-3aH-cyclopenta[a]chrysen-3a-yl)((S)- 2-(5-(pyridin-2-yl)-lH-imidazol-2-yl)pyrrolidin-l-yl)methanone:
  • Step 3 2, 2-dimethyl-4-oxo-4-(((lR, 3aS, 5aR, 5bR, 7aR, 9S, llaR l bR, 13aR, 13bR ) -5a, 5b, 8, 8, lla-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-(5-(pyridin-2-yl)-lH-imidazol-2-yl) pyrrolidine-1 -carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)butanoic acid:
  • reaction mixture was diluted with EtOAc, washed with water followed by IN HC1, water and brine solution. The residue was dried over Na 2 S0 4 , the solvent was evaporated under vacuum and purified by silica gel column (100-200 mesh, elution 5% MeOH/DCM) to afford the title compound (wt: 0.15 g) as an off white solid.
  • Example 63 Preparation of 3,3-dimethyl-5-oxo-5-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR, 13aR,13bR)-5a,5b,8,8,l la-pentamethyl-l-(l-methylcvclopropyl)-3a-((S)-2-(5-(pyridin-2-yl)- lH-imidazol-2-yl)pyrrolidine-l-carbonyl)icosahvdro-lH-cvclopentaralchrvsen-9-yl)oxy) pentanoic acid:
  • Example 64 Preparation of 5-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2- (5-(3-fluorophenyl)-lH-imidazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b,8,8 J la-pentamethyl-1- (l-methylcyclopropyl)icosahydro-lH-cyclo ⁇
  • Example 65 Preparation of 4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2- (5-(3-fluorophenyl)-lH-imidazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b,8,8 J la-pentamethyl-1- (l-methylcvclopropyl)icosahvdro-lH-cvclopentaralchrvsen-9-yl)oxy)-2,2-dimethyl-4-oxo butanoic acid:
  • Example 66 Preparation of 4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2 -(5-(4-(2-methoxyethoxy)phenyl)- lH-imidazol-2-yl)pyrrolidine- l-carbonyl)-5a,5b,8,8 J la- pentamethyl-l-(l-methylcvclopropyl)icosahvdro-lH-cvclopentaralchrvsen-9-yl)oxy)-2,2- dimethyl-4-oxobutanoic ac
  • Example 68 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-l-(l-methylcvclopropyl)-3a-(4-(5-phenyl-lH- imidazol-2-yl)piperidine-l-carbonyl)icosah ⁇
  • Example 69 Preparation of 4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-(4-(3- isopropyl-5-methyl-4H-l,2,4-triazol-4-yl)piperidine-l-carbonyl)-5a,5b,8,8 J la-pentamethyl- l-(l-methylcvclopropyl)icosahvdro-lH-cvclopentaralchrvsen-9-yl)oxy)-2,2-dimethyl-4-oxo butanoic acid:
  • Example 70 Preparation of 5-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2- (5-(4-fluorophenyl)-lH-imidazol-2-yl)pyrrolidine-l-carbonyl)-5a,5b,8,8 J la-pentamethyl-1- (l-methylcvclopropyl)icosahvdro-lH-cvclopentaralchrvsen-9-yl)oxy)-3,3-dimethyl-5-oxo pentanoic acid:
  • Example 71 Preparation of 4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((lR, 3R,5S)-3-(3-isopropyl-5-methyl-4H-l,2,4-triazol-4-yl)-8-azabicvclor3.2.11octane-8- carbonyl)-5a,5b,8,8 J la-pentamethyl-l-(l-methylcyclopropyl)icosahydro-lH-cyclopentaral chrysen-9-yl)oxy)-2,2-dimethyl-4-oxobutanoic acid:
  • Example 72 Preparation of 5-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((lR, 3R,5S)-3-(3-isopropyl-5-methyl-4H-l,2,4-triazol-4-yl)-8-azabicvclor3.2.11octane-8- carbonyl)-5a,5b,8,8 J la-pentamethyl-l-(l-methylcyclopropyl)icosahydro-lH-cyclopentaral chrysen-9-yl)oxy)-3,3-dimethyl-5-oxopentanoic acid:
  • MT-2 cells were infected with required number of TCID50's of HIV-1 strains, for e.g., 92HT599.
  • the infected cells were plated at the concentration of 30, 000 cells per well in a 96 well plate.
  • Test compound was added to the 96 well plate with the final concentration of DMSO (vehicle - not more than 1%).
  • the plates are then incubated in a C0 2 incubator for -96 hours ( ⁇ 4 days) 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 estimated index for antiviral activity of the compound. Percent inhibition was calculated with reference to control values (vehicle controls).
  • Example 54 Evaluation of compounds cyto-toxicity (MTT Assay):
  • test item was administered through oral route to animals (rat/ mice) at 30mg/kg dose in a suitable vehicle at 10 ml/ kg dose volume.
  • Blood samples ( ⁇ 50 uL at each time point) were collected from retro-orbital plexus using K3 EDTA as anticoagulant in eppend;orf tubes at defined time intervals under light ether anesthesia. The samples were centrifuged at 3500xg to separate plasma and stored at -80°C until analysis.
  • Test samples were analyzed using LC-MS-MS after developing fit- for-purpose method for each of test compound.
  • 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.

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Abstract

La présente invention concerne de nouveaux dérivés substitués par proline bétulinique, des composés apparentés et des compositions pharmaceutiques utiles dans le traitement thérapeutique de maladies virales et en particulier de maladies induites par le VIH. Formule (I)
PCT/IB2015/054784 2014-06-26 2015-06-25 Nouveaux dérivés substitués par proline bétulinique utilisés comme inhibiteurs du vih Ceased WO2015198263A2 (fr)

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US15/321,970 US20170129916A1 (en) 2014-06-26 2015-06-25 Novel betulinic proline imidazole derivatives as hiv inhibitors

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Cited By (4)

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US9868758B2 (en) 2014-06-30 2018-01-16 Hetero Labs Limited Betulinic proline imidazole derivatives as HIV inhibitors
US10370405B2 (en) 2015-03-16 2019-08-06 Hetero Labs Limited C-3 novel triterpenone with C-28 amide derivatives as HIV inhibitors
US10533035B2 (en) 2015-02-09 2020-01-14 Hetero Labs Ltd. C-3 novel triterpenone with C-17 reverse amide derivatives as HIV inhibitors
JPWO2022270628A1 (fr) * 2021-06-25 2022-12-29

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TW200628161A (en) * 2004-11-12 2006-08-16 Panacos Pharmaceuticals Inc Novel betulin derivatives, preparation thereof and use thereof
US20120101098A1 (en) * 2009-05-15 2012-04-26 Kuo-Hsiung Lee 3,28-disubstituted betulinic acid derivatives as anti-hiv agents
US9067966B2 (en) * 2009-07-14 2015-06-30 Hetero Research Foundation, Hetero Drugs Ltd. Lupeol-type triterpene derivatives as antivirals
RS54239B1 (sr) * 2010-06-04 2015-12-31 Bristol-Myers Squibb Company C-28 amidi modifikovanih derivata c-3 betulinske kiseline kao inhibitori sazrevanja hiv-a

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9868758B2 (en) 2014-06-30 2018-01-16 Hetero Labs Limited Betulinic proline imidazole derivatives as HIV inhibitors
US10533035B2 (en) 2015-02-09 2020-01-14 Hetero Labs Ltd. C-3 novel triterpenone with C-17 reverse amide derivatives as HIV inhibitors
US11034718B2 (en) 2015-02-09 2021-06-15 Hetero Labs Limited C-3 novel triterpenone with C-17 reverse amide derivatives as HIV inhibitors
US10370405B2 (en) 2015-03-16 2019-08-06 Hetero Labs Limited C-3 novel triterpenone with C-28 amide derivatives as HIV inhibitors
JPWO2022270628A1 (fr) * 2021-06-25 2022-12-29
WO2022270628A1 (fr) * 2021-06-25 2022-12-29 国立大学法人 大分大学 Dérivé de polyhétéroazole substitué par hétérocycloalkyle utilisé en tant que médicament pour le traitement et/ou la prévention d'une maladie infectieuse à virus rs

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