WO2017017630A1 - Novel betulinic substituted amide derivatives as hiv inhibitors - Google Patents

Novel betulinic substituted amide derivatives as hiv inhibitors Download PDF

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WO2017017630A1
WO2017017630A1 PCT/IB2016/054505 IB2016054505W WO2017017630A1 WO 2017017630 A1 WO2017017630 A1 WO 2017017630A1 IB 2016054505 W IB2016054505 W IB 2016054505W WO 2017017630 A1 WO2017017630 A1 WO 2017017630A1
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carbonyl
chrysen
oxy
cyclopenta
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PCT/IB2016/054505
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French (fr)
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Parthasaradhi Reddy BANDI
Rathnakar Reddy KURA
Panduranga Reddy ADULLA
David Krupadanam GAZULA LEVI
Venkati MUKKERA
Sudhakar NEELA
Vl Subrahmanyam LANKA
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Hetero Research Foundation
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J63/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton has been modified by expansion of only one ring by one or two atoms
    • C07J63/008—Expansion of ring D by one atom, e.g. D homo steroids

Definitions

  • the present invention relates to novel betulinic substituted amide 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 2016001820 describes novel betulinic proline imidazole derivatives as HIV inhibitors
  • WO 2015198263 describes novel betulinic proline substituted derivatives as hiv inhibitors
  • WO 2014/105926 describes novel betulinic acid proline derivatives as HIV inhibitors
  • WO 2013/160810 describes novel betulinic acid derivatives as HIV inhibitors
  • molecular pharmaceutics, volume: 9, issue: 11, pages: 3147-3159, 2012 describes BBA, a derivative of 23-hydroxybetulinic acid, potently reverses ABCB 1 -mediated drug resistance in vitro and in vivo
  • European Journal of Medicinal Chemistry, Volume: 46, Issue: 6, Pages: 2490-2502, 2011 describes synthesis and antiproliferative evaluation of 23-hydroxybetulinic acid derivatives
  • WO 2011/007230 describes lupeol-type triterpene derivatives as anti
  • WO 2013/148067 describes preparation of 3,28-disubstituted betulinic acid derivatives as anti-HIV agents;
  • WO 2012/154554 describes fatty acid triterpene derivatives and their uses;
  • WO 2010/032123 describes preparation of triterpenoid compounds for pharmaceutical use;
  • WO 2008/138200 describes preparation of lupane derivatives as NMDA and MC receptor antagonists exhibiting neuroprotective and memory enhancing activities;
  • WO 2008/127364 describes preparation of betulinic acid derivatives for use in antiviral and anticancer pharmaceutical compositions;
  • WO 2008/097341 describes antiviral compounds and use thereof;
  • WO 94/26725 describes preparation of lupane antiviral agents;
  • WO 2006/053255 describes novel Betulin derivatives, preparation thereof & use thereof;
  • CN 103342729 describes caffeoyl-substituted pentacyclic triterpene derivative useful in treatment of various diseases and its preparation;
  • the present invention relates to the com ounds of the formula (I):
  • n be substituted or unsubstituted alkyl
  • R b can be hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl
  • R 2 can be hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted aminoacids, substituted or unsubstituted alkoxy or substituted or unsubstituted cycloalkyl;
  • X can be absent, O, S, CH 2 or NR a (wherein R a can be H, C(0)R c , C(S)R C , substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, or R a is with their adjacent N and Carbon together form N-contained heterocycle (Preferably, pyrrolidine, piperdine, piperzine, or morpholine));
  • Y can be C(O), C(S) or CR e R f (wherein R e and R f are independently selected from H, substituted or unsubstituted alkyl);
  • 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 are taken together with the carbon atoms to which they are attached to form a bond or R 3 and R 4 are taken together with the carbon atoms to which they are attached to form cycloalkyl or R 3 and R4 are taken together with the carbon atoms to which they are attached to form epoxide;
  • R5 and R c can be independently selected from H, C0 2 R d (wherein R d is H, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl), or substituted or unsubstituted alkyl;
  • R 6 and R 8 can be independently selected from H, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl;
  • R 7 can be substituted or unsubstituted alkyl or substituted or unsubstituted cycloalkyl
  • R 6 and R 7 can be taken together with the nitrogen and carbon to which they are attached to form substituted or unsubstituted 4-7 membered heterocyclyl, substituted or unsubstituted 4-7 membered bridged heterocyclyl wherein the substituents are alkyl, heterocyclyl or -O- heterocyclyl;
  • R 7 and R 8 are taken together with the carbon atom to which they are attached to form 3-
  • Zi, Z 2 and Z 3 can be independently selected from H, hydroxy, halo, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkylsulfonyl, substituted or unsubstituted alkoxy, substituted or unsubstituted alkoxylalkoxy substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, substituted or unsubstituted heterocyclylalkyl, substituted or unsubstituted aminoalkyl, substituted or unsubstituted hydroxyalkyl, -O-heterocyclyl, substituted or unsubstituted heteroaryl; wherein the substituents are aryl, alkyl, alkoxy or alkoxylalkoxy including pharmaceutically acceptable salts, pharmaceutically acceptable solvates, pharmaceutically acceptable hydrates, tautomers, stereoisomers,
  • prodrugs of the compounds of the formula (I), including ester prodrugs are also contemplated.
  • R 7 is alkyl (preferably methyl, isobutyl).
  • ⁇ is CR e R f or -C(O)-; wherein R e and R f are hydrogen.
  • the said 4-10 membered heterocyclyl is morpholinyl, pyrrolidinyl, piperidinyl, piperizinyl, azepane and 8-azabicyclo[3.2.1]octane, thiomorpholine 1,1 -dioxide and thiomorpholinyl.
  • the said 4-10 membered heteroaryl is pyrrole, imidazoylyl, pyrazolyl and lH-pyrrolo[2,3-b]pyridine.
  • prodrugs of the compounds of the formula (IA), including ester prodrugs are also contemplated.
  • Ri, R 2 , R 3 , R4, R5, R 8 , 'X', ⁇ , Zi, Z 2 and Z 3 are as defined above including pharmaceutically acceptable salts, pharmaceutically acceptable solvates, pharmaceutically acceptable hydrates, tautomers, stereoisomers, ester prodrugs, or combination thereof.
  • a method for treating mammals infected with a virus, especially wherein said virus is HIV comprising administering to said mammal an antiviral effective amount of a compound which is selected from the group of compounds of formula (I), and one or more pharmaceutically acceptable carriers, excipients or diluents.
  • the compound of formula (I) can be administered in combination with an antiviral effective amount of another AIDS treatment agent selected from the group consisting of: (a) an AIDS antiviral agent or (b) an anti-infective agent
  • Another embodiment of the present invention is a pharmaceutical composition
  • a pharmaceutical composition comprising one or more compounds of formula (I), and one or more pharmaceutically acceptable carriers, excipients, and/or diluents; and optionally in combination with another AIDS treatment agent selected from the group consisting of: AIDS antiviral agent or anti- infective agent
  • the present invention also encompasses the method(s) of preparation of intermediates used in the preparation of compound of formula (I).
  • 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 a retroviral infection genetically related to AIDS.
  • Anti HIV inhibitory potential of the compounds of present invention may be demonstrated by any one or more methodologies known in the art, such as by using the assays described in Mossman T, December 1983, Journal of immunological methods, 65 (1- 2), 55-63 and SPC Cole, cancer chemotherapy and Pharmacology, 1986, 17, 259-263.
  • the present invention provides betulinic substituted amide 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.
  • halogen or halo includes fluorine, chlorine, bromine, or iodine.
  • alkyl refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation, having from one to eight carbon atoms, and which is attached to the rest of the molecule by a single bond, e.g., methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), isobutyl, n-butyl, n-pentyl, and 1,1-dimethylethyl (t-butyl).
  • 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.
  • 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.
  • alkyl sulfonyl refers to a group designates an "alkyl-S0 2 -" group, wherein alkyl is as defined above.
  • alkyl sulfonyl groups of the invention include methyl sulfonyl, ethyl sulfonyl and propyl sulfonyl.
  • aryl refers to an aromatic radical having from 6 to 14 carbon atoms such as phenyl, naphthyl, tetrahydronapthyl, indanyl, and biphenyl.
  • arylalkyl refers to an aryl group as defined above directly bonded to an alkyl group as defined above, e.g., -CH 2 C 6 H 5 and -C 2 H 5 C 6 H 5 .
  • 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.
  • nitrogen atom may be optionally quaternized;
  • heterocyclic ring radicals include, but are not limited to, , tetrahydroisouinolyl, piperidinyl, piperazinyl, azapanyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxoazepinyl, azepinyl, pyrrolyl, 4-piperidonyl, pyrrolidinyl, pyrazinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, decahydroisoquinolyl, tetrahydrofurtyl, tetrahydropyranyl, thienyl, benzothienyl, thiamorpholinyl, thiamorpholinyl sulfoxide, thiamorpholinyl sulfone, di
  • bridged heterocyclyl refers to unsaturated or saturated bridged cyclic system that is not aromatic. Such a system may contain isolated or conjugated unsaturation, but not aromatic or heteroaromatic rings in its core structure (but may have aromatic substitution thereon). And at least one ring in the system is inclusive of one or more heteroatoms, wherein each ring in the system contains 3 to 7 ring members, e.g., 1 to 10 carbon atoms and 1 to 3 heteroatoms selected from N, O, P or S, wherein the S or P is optionally substituted with one or more oxo to provide the group SO or S0 2 , PO or PO 2 .
  • bridged heterobicyclic ring system examples include (lR,5S)-8- azabicyclo[3.2.1]octane, (lR,5S)-3-azabicyclo[3.1.0]hexane or (lR,5S)-3,8- diazabicyclo[3.2.1]octane.
  • heterocyclylalkyl refers to a heterocyclic ring radical directly bonded to an alkyl group.
  • the heterocyclylalkyl radical may be attached to the main structure at any carbon atom in the alkyl group that results in the creation of a stable structure.
  • 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.
  • heteroaryl ring include, but are not limited to imidazolyl, pyrazolyl, pyrrolyl, lH-pyrrolo[2,3-b]pyridinyl, tetrazoyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolinyl, oxazolidinyl, triazolyl, isoxazolyl, isoxasolidinyl, thiazolyl, thiazolinyl, thiazolidinyl, isothiazolyl, quinuclidinyl, isothiazolidinyl, indolyl, isoindolyl, indolinyl, isoindolinyl, quinolyl
  • prodrug means a compound that is transformed in vivo to yield a compound of formula (1) 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.
  • terapéuticaally 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), 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.
  • 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 I 101 each at final concentration of virus equivalent to 2 ng of p24 per ml. The infected cells will be incubated at 37 C and 5% C02 incubator for 2 hours. After 2hrs the cells will be pelleted at 350 g for 10 min, supernatant will be discarded and cell will be held with RPMI 1640 containing 10% FBS.
  • the cells will be resuspended in the same medium with increasing concentrations of Epap-1 peptides and will be incubated for 96 hours. The cells will be supplemented with peptides at every 24 hours. The supernatants will be collected after 96 hours and analyzed using P24 antigen capture assay kit (SAIC Fredrick). The infection in the absence of Epap-1 will be considered to be 0% inhibition Azidothymidine (AZT) will be taken as positive control.
  • 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, FHV 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
  • the compounds of the present invention can obtain more advantageous effects than additive effects in the prevention or treatment of the above diseases when using suitably in combination with the available drugs. Also, the administration dose can be decreased in comparison with administration of either drug alone, or adverse effects of co administrated drugs other than antiviral can be avoided or declined.
  • 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 3. 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.
  • the compounds of the present invention can also contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds.
  • the present invention also embraces isotopically-labeled variants of the present invention which are identical to those recited herein, but for the fact that one or more atoms of the compound are replaced by an atom having the atomic mass or mass number different from the predominant atomic mass or mass number usually found in nature for the atom. All isotopes of any particular atom or element as specified are contemplated within the scope of the compounds of the invention, and their uses.
  • Exemplary isotopes that can be incorporated in to compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, sulfur, fluorine, chlorine and iodine, such as H ("D"), 3 H, n C, 13 C, 14 C, 13 N, 15 N, 15 0, 17 0, 18 0, 32 P, 33 P, 35 S, 18 F, 36 C1, 123 I and 125 I.
  • Particular isotopes are -CD 3 or -C(D 2 )-.
  • Isotopically labeled compounds of the present inventions can generally be prepared by following procedures analogous to those disclosed in the Schemes and/or in the Examples herein below, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.
  • Compounds of the present invention can be synthesized from naturally occurring Betulinic acid or betulinal.
  • Key intermediates required for synthesizing analogues are either commercially available, or can be prepared by the methods published in the literature.
  • the key intermediates in the present invention were prepared by modifying the procedures published in Journal of organic chemistry 2013, 78, 1463-1470; Organic Letters, 2009, 11, 3270-3273; Organic Letters, 2007, 9, 3675-3678; Bio Organic Medicinal Chemistry Letters, 2011, 21, 562-568.
  • the compounds of formula 5 (R 7 , R 8 , Zi, Z 2 and Z 3 are same as defined above) can be prepared as described in Scheme 1.
  • the nitro compound of formula 1 can be reacted with the aldehydes of formula 2 followed by substituted N-contained heterocycles of formula 3 to give the compounds of formula 4 in the presence of base solutions such as sodium hydroxide (NaOH), potassium hydroxide (KOH) or the like.
  • the compounds of formula 4 can be reduced with reducing agents such as raney nickel, palladium in carbon (Pd/C) or the like to give the compounds of formula 5 in the presence of suitable solvents such as methanol (MeoH), ethylacetate (EtOAc) or the like.
  • the compounds of formula 12 & 14 ( ' ⁇ -—-'' , Zi, Z 2 and Z 3 are same as defined above) can be prepared as described in scheme 2.
  • the prolinal compounds of formula 6 can be converted to N-protected compounds of formula 7 in the presence of protecting groups such as tert-butyloxy carbonyl (BOC), benzyl, carboxybenzyl (cbz) or the like in the presence of solvents such as dichloromethane (DCM), ⁇ , ⁇ -dimethyl formamide (DMF), tetrahydrofuran (THF) or the like.
  • protecting groups such as tert-butyloxy carbonyl (BOC), benzyl, carboxybenzyl (cbz) or the like in the presence of solvents such as dichloromethane (DCM), ⁇ , ⁇ -dimethyl formamide (DMF), tetrahydrofuran (THF) or the like.
  • DCM dichloromethane
  • DMF ⁇ , ⁇
  • the compounds of formula 7 can be converted to compounds of formula 8 in the presence of bases such as lithium aluminum hydride (LAH), sodiumborohydride (NaBH 4 ), boron in tetrahydrofuran or the like in the solvents such as tetrahydrofuran (THF), ether, ethylacetate (EtOAc) or the like.
  • bases such as lithium aluminum hydride (LAH), sodiumborohydride (NaBH 4 ), boron in tetrahydrofuran or the like in the solvents such as tetrahydrofuran (THF), ether, ethylacetate (EtOAc) or the like.
  • the compounds of formula 8 can be converted to O- protected compounds of formula 9 in the presence of protecting groups such as p-toluene sulfonyl chloride, benzyl, i-Butyldimethylsilyl (TBDMS), benzoyl or the like in the solvents such as dichloromethane (DCM), tetrahydrofuran (THF) or the like.
  • protecting groups such as p-toluene sulfonyl chloride, benzyl, i-Butyldimethylsilyl (TBDMS), benzoyl or the like in the solvents such as dichloromethane (DCM), tetrahydrofuran (THF) or the like.
  • the compounds of formula 9 coupled with compounds of formula 10 can be converted to compounds of formula 11 in the presence of solvents such as 1,4-dioxane, tetrahydrofuran (THF), toluene or the like in presence of without base and with bases such as sodium hydroxide (NaOH), sodium bicarbonate (NaHC0 3 ), sodium carbonate (Na 2 C0 3 ) or the like.
  • solvents such as 1,4-dioxane, tetrahydrofuran (THF), toluene or the like
  • bases such as sodium hydroxide (NaOH), sodium bicarbonate (NaHC0 3 ), sodium carbonate (Na 2 C0 3 ) or the like.
  • the compounds of formula 11 can be converted to compounds of formula 12 in the presence of deprotecting reagents such as HC1, trifluoro acetic acid (TFA) or the like in the solvents such as dichloromethane (DCM), dioxane or the like.
  • the compounds of formula 7 can be coupled with the compounds of formula 10 to give the compounds of formula 13 in the presence of l-Ethyl-3- (3-dimethylaminopropyl)carbodiimide (EDCI), N-Hydroxybenzotriazole (HOBt) or the like in the solvents such as ⁇ , ⁇ -dimethyl formamide (DMF), N,N-Diisopropylethylamine (DIPEA) or the like.
  • deprotecting reagents such as HC1, trifluoro acetic acid (TFA) or the like in the solvents such as dichloromethane (DCM), dioxane or the like.
  • the compounds of formula 7 can be coupled with
  • the compounds of formula 13 can be deprotected in the presence deprotecting reagents such as HC1, trifluoro acetic acid (TFA) or the like in the solvents such as dichloromethane (DCM), dioxane or the like to give the compounds of formula 14.
  • deprotecting reagents such as HC1, trifluoro acetic acid (TFA) or the like in the solvents such as dichloromethane (DCM), dioxane or the like to give the compounds of formula 14.
  • the compounds of formula (I) (wherein, Ri, R 2 , R 3 , R4, R5, R 6 , R7, X, Zi, Z 2 and Z 3 are same as defined above) can be prepared as described in Scheme 3.
  • the compounds of formula 15 (as described in our PCT publication WO 2013/160810) can be converted to the halide compounds of formula 16 in the presence of halogenating agents such as thionyl chloride (SOCl 2 ), oxalyl chloride (COCl 2 ), phosphorous bromide, phosphorous oxy bromide, phosphorous pentachloride, phosphorous tribromide, phosphorous pentabromide or the like in the solvents such as benzene, toluene, dichloromethane (DCM) or the like.
  • halogenating agents such as thionyl chloride (SOCl 2 ), oxalyl chloride (COCl 2 ), phosphorous bromide, phosphorous oxy bro
  • the compounds of formula 16 can be converted to amide compounds of formula 17 with their corresponding amine compounds of formula 5 in the presence of bases such as triethylamine (TEA), N,N- diisopropylethylamine (DIPEA), pyridine or the like in the solvents such as N,N- di.methylfonnam.ide (DMF), dichloromethane (DCM), tetrahydrofuran (THF) or the like.
  • bases such as triethylamine (TEA), N,N- diisopropylethylamine (DIPEA), pyridine or the like in the solvents such as N,N- di.methylfonnam.ide (DMF), dichloromethane (DCM), tetrahydrofuran (THF) or the like.
  • C28-amide compounds of formula 17 can also be prepared by using suitable coupling agents such as l-ethyl-3-(3-dimethylaminopropyl) carbodiimide) hydrochloride (EDCI), (l-[Bis(dimethylamino)methylene]-lH-l,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate) (HATU), 1- Hydroxybenzotriazole (HOBt) monohydrate or the like with suitable bases such as tri ethyl amine (TEA), ⁇ , ⁇ -diisopropylethy famine (DIPEA), pyridine or the like with nicotinykhloride or benzoylchioride in the solvents such as N,N- dimethylfomi amide (DM F), dichloromethane (DCM), tetrahydrofuran (TBF) or the like.
  • suitable coupling agents such as l-eth
  • the compounds of formula 17 can be deprotected in the presence of bases such as potassium carbonate, sodium hydroxide, ammonia or the like in the solvents such as methanol: tetrahydrofuran, methanol: water, methanol or the like to give the C3 hydroxy compounds of formula 18.
  • the C3 hydroxy compounds of formula 18 can be reacted with corresponding acid anhydrides, half protected diacids or their mixed anhydrides or acid chlorides to give the corresponding acid compounds of the present invention formula (1) in the presence a bases such as tri ethyl amine (TEA), 4-dimethylaminopyridine, diisopropyl. ethyl mine or pyridine or the like in the solvents such as dichloromethane (DCM), toluene, ethylacetate (EtOAc), tetrahydrofuran (THF) or the like.
  • bases such as potassium carbonate, sodium hydroxide, ammonia or the like in the solvents such as
  • DIPEA ⁇ , ⁇ -Diisopropylethylamine
  • °C degree Celsius
  • ⁇ delta
  • ppm parts per million
  • % percentage
  • DMSO-d 6 Deuterated DMSO
  • d Doublet
  • dd Doublet of doublet
  • EtOH Ethanol
  • EtOAc Etthyl acetate
  • g or gr gram
  • H or H 2 Hydrogen
  • HC1 Hydrochloric acid
  • HATU (0-(7-Azabenzotriazol-l-yl)- ⁇ , ⁇ , ⁇ ', ⁇ '-tetramethyluroniumhexafluoro phosphate); Hz (Hertz); HPLC (High-performance liquid chromatography); mmol (Milli mol); M (Molar); ml (Millilitre); mg (Milli gram); m (Multiplet); mm (Millimetre); MHz (Megahertz); ESI-MS (Electron spray Ionization Mass spectra); min (Minutes); mM (Milli molar); NaOH (Sodium hydroxide); N 2 (Nitrogen); NMR (Nuclear magnetic resonance spectroscopy); S (Singlet); TEA (Triethyl amine); TLC (Thin Layer Chromatography); THF (Tetrahydrofuran); tert (Tertiary), t (Triplet); IC (Inhibitory concentration),
  • Step 2 Synthesis of 2 -methyl- l-morpholinopropan-2 -amine:
  • step 1 4-(2-methyl-2-nitropropyl)morpholine (step 1) in methanol (20 mL) then added Raney Nickel (1.7 g, 29.79 mmol, 1.4 eq) treated with hydrogen (50 psi) at room temperature for about 1 hour. After completion of the reaction (Monitored by TLC), the reaction mixture was filtered through celite bed and the filtrate was concentrated to afford the desired product (3.2 g, yield: 95.0%) as a light yellow solid.
  • Step 1 Synthesis of tert-butyl 4-benzylpiperazine-l-carboxylate:
  • step 1 To the stirred solution of tert-butyl 4-benzylpiperazine-l-carboxylate (step 1, 6.5 g, 18.5 mmol, 1.0 eq) in 1,4-dioxane in HCl and stirred at room temperature for about 3 hours. The reaction mixture was evaporated under reduced pressure to afford the desired product (4.0 g) as a white solid.
  • Step 3 Synthesis of 1 -benzyl-4-(2-methyl-2-nitropropyl)piperazine :
  • step 2 To the stirred solution of 1-benzylpiperazine (step 2, 4.0 g, 29.1 mmol, 1.0 eq), 2- nitropropane (2.8 mL, 29.1 mmol, 1.0 eq) are cooled with ice bath, then added formaldehyde (37%) (2.5 mL, 70.5 mmol, 3.0 eq) and NaOH solution (5.0 mL, 0.5 mol/L) are slowly dropwise at 10 °C. Then the reaction mixture was stirred for about 1 hour at 25 °C and about 1 hour at 50 °C. The solution is treated with water, ether and aqueous phase is extracted with water.
  • Step 4 Synthesis of 1 -(4-benzylpiperazin-l -yl)-2-methylpropan-2-amine:
  • step 3 To the stirred solution of l-benzyl-4-(2-methyl-2-nitropropyl)piperazine (step 3, 2.0 g, 7.3 mmol, 1.0 eq) in Methanol (30 mL), then added Raney Nickel (0.200 g, 4.8 mmol, 0.6 eq) and treated with hydrogen (50 psi) at room temperature for about 12 hours. The reaction mixture was filtered through celite and filtrate was concentrated to obtain the desired product (1.2 g, yield: 65%) as a white powder.
  • Step 1 Synthesis of tert-butyl (R)-3-hydroxypyrrolidine-l-carboxylate:
  • Step 2 Synthesis of tert-butyl (R)-3- 2-methoxyethoxy)pyrrolidine-l-carboxylate:
  • Step 4 Synthesis of (R)-l-(3-(2-methoxyethoxy)pyrrolidin-l-yl)-2-methylpropan-2-amine:
  • step 3 To a stirred solution of (R)-3-(2-methoxyethoxy)-l-(2-methyl-2-nitropropyl) pyrrolidine (step 3, 2.0 g, 8.13 mmol) in methanol (20 ml) was added 10% Pd/C (0.20 g) and purged with nitrogen. The reaction mixture was stirred for about 12 hours under H 2 atmosphere. After completion of the reaction (monitored by TLC), the reaction mixture was filtered and filtrate was concentrated under reduced pressure to give the desired product. Next reaction was carried out without any further purification.
  • Step 1 Synthesis of tert-butyl 4-oxopiperidine-l -carboxylate:
  • Step 2 Synthesis of tert-butyl 4-hydroxypiperidine-l-carboxylate:
  • step 1 To a stirred solution of tert-butyl 4-oxopiperidine-l-carboxylate (step 1, 12.95 g, 65 mmol) in MeOH (130 ml), was slowly added NaBH 4 (12.3 g, 325 mmol) over about 15 minutes (Care - vigorous effervescence) at 0 °C. The reaction mixture was allowed to stir at room temperature for about 5 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by addition of saturated NH 4 C1 solution at 0°C. The solution was extracted with EtOAc (2x150 mL), the combined organic phases were washed with brine, dried over Na 2 S0 4 and concentrated under reduced pressure to give the crude product.
  • EtOAc 2x150 mL
  • Step 3 Synthesis of tert-butyl 4-(2-methoxyethoxy)piperidine-l-carboxylate:
  • Step 4 Synthesis of 4-(2-methoxyethoxy)-l -(2-methyl-2-nitropropyl)piperidine:
  • Step 5 Synthesis of 1 -(4-(2-methoxyethoxy)piperidin-l -yl)-2-methylpropan-2 -amine:
  • step 4 To a stirred solution of 4-(2-methoxyethoxy)-l-(2-methyl-2-nitropropyl)piperidine (step 4, 1.6 g, 6.6 mmol) in methanol (20 ml) was added 10% Pd/C (0.20 g) and purged with nitrogen. The reaction mixture was stirred for about 12 hours under H 2 atmosphere. After completion of the reaction (monitored by TLC), the reaction mixture was filtered and filtrate was concentrated under reduced pressure to give the desired product (1.3 g, yield: 85%) as an oil.
  • Step 2 Synthesis of 4-ethoxy-l -(2-methyl-2-nitropropyl)-4-phenylpiperidine:
  • Step 3 Synthesis of 1 -(4-ethoxy-4-phenylpiperidin-l -yl)-2-methylpropan-2-amine:
  • step 2 To a stirred solution of 4-ethoxy-l-(2-methyl-2-nitropropyl)-4-phenylpiperidine (step 2, 0.3 g, 0.9 mmol) in methanol (8 ml) was added 10% Pd/C (0.03 g) and purged with nitrogen. The reaction mixture was stirred for about 12 hours under H 2 atmosphere. After completion of the reaction (monitored by TLC), the reaction mixture was filtered and filtrate was concentrated under reduced pressure to give the desired product (0.15 g, yield: 72.5%) as a gummy oil. Next reaction was carried out without any further purification.
  • Step 1 Synthesis of tert-butyl (R)-3 -(benz loxy pyrrolidine- 1-carboxylate:
  • Step 2 Synthesis of ' (R)-3-(benzyloxy)pyrrolidine: , ⁇
  • step 1 To the stirred solution of tert-butyl (R)-3-(benzyloxy)pyrrolidine-l-carboxylate (step 1) in 1,4-dioxane in HC1 and stirred at room temperature for about 3 hours. The reaction mixture evaporated under reduced pressure to afford the desired product (2.0 g) as a white solid.
  • Step 3 Synthesis of (R)-3 -(benzyloxy)-l -(2-methyl-2-nitropropyl)pyrrolidine :
  • step 2 To the stirred solution of (R)-3-(benzyloxy)pyrrolidine (step 2, 2.0 g, 11.29 mmol, 1.0 eq) and 2-nitropropane (1.2 mL, 13.55 mmol, 1.2 eq) are cooled with ice bath then added formaldehyde (37%) (2.1 mL, 27.09 mmol, 2.4 eq) and NaOH solution (2.0 mL, 0.5 mol/L) are slowly dropwise at 10 °C. Then the reaction mixture was stirred for about 1 hour at 25 °C and about 1 hour at 50 °C. The solution is treated with water, ether and aqueous phase is extracted with water.
  • Step 4 Synthesis of (R)-l -(3-(benzyloxy)pyrrolidin-l -yl)-2-methylpropan-2-amine:
  • step 3 To the solution of (R)-3-(benzyloxy)-l-(2-methyl-2-nitropropyl)pyrrolidine (step 3, 2.0 g, 7.19 mmol, 1.0 eq) in Methanol (30 mL) then added Raney Nickel (1.0 g, 35.89 mmol, 1.4 eq) and treated with hydrogen (50 psi) and stirred at room temperature for about 1 hour. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through celite and filtrate was concentrated to afford the desired product (1.2 g, yield: 70%) as a white powder.
  • Step 1 Synthesis of tert-butyl piperazine-1 -carboxylate:
  • Step 2 Synthesis of tert-butyl 4-(methylsulfonyl)piperazine-l-carboxylate:
  • Triethyl amine 22 ml, 160 mmol, 3.0 eq
  • methane sulphonyl chloride 4.5 ml, 59 mmol, 1.1 eq
  • DMAP 0.66 g, 5.4 mmol, 0.1 eq
  • step 1 a solution of tert-butyl piperazine- 1-carboxylate
  • step 2 a solution of tert-butyl piperazine- 1-carboxylate
  • step 2 A solution of tert-butyl 4-(methylsulfonyl)piperazine- 1-carboxylate (step 2, 10 g, 38.0 mmol, 1.0 eq) in 1,4-dioxane.HCl (100 mL, 10 vol.) was stirred at room temperature for about 4 hours.
  • Step 4 Synthesis of 1 -(2-methyl-2-nitropropyl)-4-(methylsulfonyl)piperazine:
  • Step 5 Synthesis of 2-methyl-l -(4-(methylsulfonyl)piperazin-l -yl)propan-2-amine:
  • Step 2 Synthesis of tert-butyl (S)-2-(4-ethylpiperazine-l -carbonyl)pyrrolidine-l -carboxylate:
  • step 1 A solution of (tert-butoxycarbonyl)-L-proline (step 1, 25.0 g, 116.82 mmol, 1.0 eq) in DMF (250 mL) was treated sequentially with HOBt (18.9 g, 140.18 mmol, 1.2 eq) and EDCI (33.4 g, 170.5 mmol, 1.5 eq) and 1 -ethylpiperazine (14.67 g, 175.23 mmol, 1.5 eq). The suspension was stirred for about 15 minutes, TEA (80 mL, 584.0 mmol, 5.0 eq) was added, and the mixture stirred for about 12 hours.
  • HOBt 18.9 g, 140.18 mmol, 1.2 eq
  • EDCI 33.4 g, 170.5 mmol, 1.5 eq
  • 1 -ethylpiperazine 14.67 g, 175.23 mmol, 1.5 eq
  • the reaction was quenched by pouring into 50 mL of ethyl acetate, water and extracting with ethylacetate, washed with water, saturated aqueous NaHC0 3 and a portion of brine. The solution was then dried over Na 2 S0 4 , filter and concentrated. The residue was purified by column chromatography on silica gel 2% MeOH and DCM to afford the desired product (5.5 g) as an off white solid.
  • Step 4 Synthesis of (S)-l -ethyl-4-(pyrrolidin-2-ylmethyl)piperazine :
  • reaction mixture was diluted with DCM and washed with water several times and brain solution and organic layer was dried with anhydrous Na 2 S0 4 , and concentrated under reduced pressure to afford the desired product (80.0 g, yield: 74.0%), which was proceed for the next step without purification.
  • Step 2 Synthesis of tert-butyl (R)-2-(pyrrolidine-l -carbonyl)pyrrolidine-l -carboxylate:
  • step 1 A solution of (tert-butoxycarbonyl)-D-proline (step 1, 5.g, 23.0 mmol, 1.0 eq) in DMF (25 mL) was treated sequentially with HOBt (6.0 g, 35.0 mmol, 1.5 eq) and EDCI ( 13.0 g, 69.0 mmol, 3.0 eq) and pyrrolidine (35.0 g, 46.0 mmol, 2.0 eq). The suspension was stirred for about 15 minutes, DIPEA (20 mL, 35.0 mmol, 1.5 eq) was added and the mixture stirred for 12 hours.
  • DIPEA 20 mL, 35.0 mmol, 1.5 eq
  • the reaction was quenched by pouring into 50 mL of ethyl acetate and water extracting with ethylacetate and washed with water, saturated aqueous NaHC0 3 , and a portion of brine. The solution was then dried over Na 2 S0 4 , filtered and concentrated. The residue was purified by column chromatography on silica gel 2% MeOH and DCM to afford the desired product (3.5 g, yield: 59.0%) as an off white solid.
  • Step 3 Synthesis of (R)-l -prolylpyrrolidine : To the stirred solution of tert-butyl (R)-2-(pyrrolidine-l-carbonyl)pyrrolidine-l- carboxylate (step 2, 3.5 g, 13.2 mmol, 1.0 eq) in 1,4-dioxane.HCl (30 mL) which was stirred at room temperature for about 4 hours. After completion of the reaction (monitored by TLC), the volatile solvent was concentrated to afford the desired compound as a brown liquid which was proceed for the next step without purification.
  • Step 1 Synthesis of tert-butyl (S)-2-((S)-2-(hydroxymethyl)pyrrolidine-l-carbonyl) pyrrolidine-1 -carboxylate:
  • reaction mixture was diluted with DCM and washed with water, IN HC1 solution, saturated NaHC0 3 solution, brine and dried over Na 2 S0 4 and concentrated under reduced pressure to give the crude product. Purification by column chromatography with EtOAc- hexane (7:3) as an eluent to afford the desired product (6.8 g, yield: 75.5%) as a thick oil.
  • Step 2 Synthesis of tert-butyl (S)-2-(((S)-2-(hydroxymethyl)pyrrolidin-l-yl)methyl) pyrrolidine-1 -carboxylate:
  • Step 3 Synthesis of tert-butyl (S)-2-(((S)-2-((benzyloxy)methyl)pyrrolidin-l-yl)methyl) pyrrolidine-1 -carboxylate:
  • Step 4 Synthesis of (S)-2-((benzyloxy)methyl)-l-(((S)-pyrrolidin-2-yl)methyl)pyrrolidine hydrochloride:
  • Step 1 Synthesis of tert-butyl (R)-3-(((R)-tetrahydrofuran-3-yl)oxy)pyrrolidine-l- carboxylate:
  • step 1 tert-butyl (R)-3-(((R)-tetrahydrofuran-3-yl)oxy)pyrrolidine-l- carboxylate (step 1, 0.9 g, 3.50 mmol) in dioxane (5 ml), was added 6N HCl in dioxane (5 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (0.67 g, yield: 100%). Next reaction was carried out without any further purification.
  • Step 3 Synthesis of tert-butyl (S)-2-((R)-3-(((R)-tetrahydrofuran-3-yl)oxy)pyrrolidine-l- carbonyl )pyrrolidine-l -carboxylate:
  • Step 4 Synthesis of (R)-l-(L-prolyl)-3-(((R)-tetrahydrofuran-3-yl)oxy)pyrrolidine hydrochloride:
  • Step 5 Synthesis of (R)-l-(((S)-pyrrolidin-2-yl)methyl)-3-(((R)-tetrahydrofuran-3-yl)oxy) pyrrolidine:
  • Step 1 Synthesis of tert-butyl 4,4-difluoropi eridine-l -carboxylate :
  • step 1 To a stirred solution of tert-butyl 4,4-difluoropiperidine- 1-carboxylate (step 1, 3.73 g, 16.87 mmol) in dioxane (20 ml), was added 6N HCl in dioxane (30 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (2.64 g, yield: 100%). Next reaction was carried out without any further purification.
  • Step 3 Synthesis of tert-butyl (S)-2-(4,4-difluoropiperidine-l-carbonyl)pyrrolidine-l- carboxylate:
  • Step 5 Synthesis of (S)-4,4-difluoro-l -(pyrrolidin-2-ylmethyl)piperidine: To a stirred suspension of LiAlH 4 (1.1 g, 27.63 mmol) in dry THF (15 ml), was slowly added a solution of (S)-4,4-difluoro-l-prolylpiperidine hydrochloride (step 4, 3.51 g, 13.81 mmol) in THF (40 ml) at 0 °C. After addition, the reaction mixture was then brought to room temperature and stirred for overnight.
  • reaction mixture was quenched by the sequential addition of 1.0 mL of H 2 0, 3 mL of 15% aq. NaOH and 3 mL of H 2 0.
  • the mixture was then poured into EtOAc (100 ml) and stirred for about 30 minutes.
  • the insoluble material was removed by filtration through celite and the solvent was removed from the filtrate by rotary evaporation and dried over Na 2 S0 4 .
  • the product was isolated by column chromatography on silica gel using MeOH: DCM (3:97) as an eluent to give the desired product (2.64 g, yield: 94%) as a solid.
  • Step 1 Synthesis of tert-butyl 3-oxopyrrolidine-l-carboxylate:
  • Step 2 Synthesis of tert-butyl 3,3-difluoropyrrolidine-l-carboxylate:
  • step 1 To a stirred solution of tert-butyl 3-oxopyrrolidine-l-carboxylate (step 1, 3.44 g, 18.59 mmol) in DCM (40 ml) under N 2 atmosphere at 0 °C, was added DAST (2.7 ml, 20.45 mmol) and stirred for about 6 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by addition of ice cold water. The solution was extracted with DCM (2x100 mL) and the combined organic phases were washed with brine, dried over Na 2 S0 4 and concentrated under reduced pressure to give the crude product.
  • DCM tert-butyl 3-oxopyrrolidine-l-carboxylate
  • step 2 To a stirred solution of tert-butyl 3,3-difluoropyrrolidine-l-carboxylate (step 2, 1.61 g, 7.77 mmol) in dioxane (15 ml), was added 6N HCl in dioxane (20 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (1.11 g, yield: 100%). Next reaction was carried out without any further purification.
  • Step 4 Synthesis of tert-butyl (S)-2-(3,3-difluoropyrrolidine-l-carbonyl)pyrrolidine-l- carboxylate:
  • Step 5 Synthesis of (S)-3 ' ,3-difluoro-l-prolylpyrrolidine hydrochloride:
  • Step 1 Synthesis of tert-butyl (S)-2-(((S)-3-fluoropyrrolidin-l-yl)methyl)pyrrolidine-l- carboxylate:
  • step 1 tert-butyl (S)-2-((4-fluoropiperidin-l-yl)methyl)pyrrolidine-l- carboxylate (step 1, 0.5 g, 1.8 mmol) in dioxane (5 ml), was added 6N HCl in dioxane (5 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (0.444 g, yield: 100%). Next reaction was carried out without any further purification.
  • Step 1 Synthesis of tert-butyl (2S,4R)-4-hydroxy-2-(pyrrolidine-l-carbonyl)pyrrolidine-l- carboxylate:
  • reaction mixture was diluted with DCM and washed with water, IN HCl solution, saturated NaHC0 3 solution, brine, dried over Na 2 S0 4 and concentrated under reduced pressure to give the crude product. Purification by column chromatography with EtOAc-hexane (1: 1) as an eluent to afford the desired product (8 g, yield: 65.5%) as a thick oil.
  • reaction mixture was quenched by the cautious addition of MeOH (70 ml) at 0 °C and allowed to stir at 60 °C for about 12 hours.
  • the reaction mixture was concentrated and the resulting residue was diluted with EtOAc and washed with water, brine, dried over Na 2 S0 4 and concentrated under reduced pressure to give the crude product. Purification by column chromatography with MeOH:DCM (5:95) as an eluent to afford the desired product (4.8 g, yield: 63.15%) as a thick oil.
  • 1H NMR 300 MHz,
  • Step 3 Synthesis of tert-butyl (2S,4R)-2-(pyrrolidin-l-ylmethyl)-4-(((R)-tetrahydrofuran-3-yl) oxy )pyrrolidine-l -carboxylate:
  • Step 4 Synthesis of (2S,4R)-2-(pyrrolidin-l-ylmethyl)-4-(((R)-tetrahydrofuran-3-yl)oxy) pyrrolidine hydrochloride:
  • Step 1 Synthesis of tert-butyl (S)-2-(2-hydroxypropan-2-yl)pyrrolidine-l-carboxylate:
  • step 1 To a stirred solution of tert-butyl (S)-2-(2-hydroxypropan-2-yl)pyrrolidine-l- carboxylate (step 1, 5.9 g, 25.7 mmol) in dioxane (50 ml), was added 6N HC1 in dioxane (50 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (4.2 g, yield: 100%). Next reaction was carried out without any further purification.
  • Step 3 Synthesis of tert-butyl (S)-2-((S)-2-(2-hydroxypropan-2-yl)pyrrolidine-l-carbonyl) pyrrolidine-1 -carboxylate:
  • reaction mixture was diluted with DCM, washed with water, saturated NaHC0 3 solution, brine and dried over Na 2 S0 4 .
  • the solvent was evaporated and to the resulting solid, was added DCM (90 ml) and stirred for about one hour and filtered. The filtrate was concentrated under reduced pressure to afford the title compound (8.0 g, yield: 97.5%) as gummy oil.
  • Step 4 Synthesis of (S)-l-(L-prolyl)-2- -hydroxypropan-2-yl)pyrrolidine hydrochloride:
  • Step 5 Synthesis of2-((S)-l-(((S)-pyrrolidin-2-yl)methyl)pyrrolidin-2-yl)propan-2-ol:
  • 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 1 To a stirred solution of tert-butyl (S)-2-(hydroxymethyl)pyrrolidine-l-carboxylate (step 1, 2.5 g, 12.4 mmol) in DCM (30 ml) under N 2 atmosphere, was added DMP (7.9 g, 18.6 mmol) at 0 °C. The reaction mixture was allowed to room temperature and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with 1 : 1 mixture of saturated NaHC0 3 and saturated Na 2 S 2 0 3 solution. The reaction mixture was extracted with DCM (2x50 ml). The combined organic layer was washed with brine, dried over anhydrous sodium sulphate and concentrated under reduced pressure to give the residue (2.1 g, 84%). Next reaction was carried out without any further purification.
  • Step 3 Synthesis of tert-butyl (S)-2-(((S)-2-(hydroxymethyl)pyrrolidin-l-yl)methyl) pyrrolidine-1 -carboxylate:
  • step 2 To a stirred solution of tert-butyl (S)-2-formylpyrrolidine-l-carboxylate (step 2, 2.1 g, 10.5 mmol) in 1,2-dichloro ethane (25 ml) under N 2 atmosphere, was added AcOH (1 ml, 15.8 mmol) at 0 °C. After stirring about 20 minutes at 0 °C, were added (S)-prolinol (1.1 g, 10.5 mmol) and followed by sodiumtriacetoxy borohydride (3.4 g, 15.8 mmol). The reaction mixture was allowed to room temperature and stirred for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with saturated NH 4 C1 solution.
  • Step 4 Synthesis of tert-butyl (S)-2-((2-formyl-lH-pyrrol-l-yl)methyl)pyrrolidine-l- carboxylate:
  • Step 5 Synthesis of tert-butyl (S)-2-((2-(pyrrolidin-l-ylmethyl)-lH-pyrrol-l-yl)methyl) pyrrolidine-1 -carboxylate:
  • reaction mixture was quenched with saturated NH 4 C1 solution.
  • the reaction mixture was extracted with DCM (2x30 ml). The combined organic layers were washed with brine, dried over anhydrous sodium sulphate and concentrated under reduced pressure to give the residue.
  • Step 6 Synthesis of (S)-2-(pyrrolidin-l-ylmethyl)-l-(pyrrolidin-2-ylmethyl)-lH-pyrrole hydrochloride:
  • Step 1 Synthesis of tert-butyl (S)-2-(4-(hydroxymethyl)piperidine-l-carbonyl)pyrrolidine-l- carboxylate:
  • the reaction was quenched by pouring into 50 mL of ethyl acetate and water, extracting with ethylacetate, washed with water and a portion of brine. The solution was then dried over Na 2 S0 4 , filter and concentrated. The residue was purified by column chromatography on silica gel 3% MeOH and DCM to afford the title compound (2.9 g, yield: 40.0%) as a brown colour liquid.
  • Step 2 Synthesis of tert-butyl (S)-2-(4-((benzyloxy)methyl)piperidine-l-carbonyl)pyrrolidine- 1-carboxylate:
  • Step 1 Synthesis of tert-butyl (2S,4R)-4-hydroxy-2-(hydroxymethyl)pyrrolidine-l- carboxylate: HO
  • Step 2 Synthesis of tert-butyl (2S,4R)-4-hydroxy-2-((2-methoxyethoxy)methyl)pyrrolidine-l- carboxylate:
  • Step 3 Synthesis of tert-butyl (2S,4S)-4-fluoro-2-((2-methoxyethoxy)methyl)pyrrolidine-l- carboxylate:
  • Step 5 Synthesis of tert-butyl (S)-2-((2S,4S)-4-fluoro-2-((2-methoxyethoxy)methyl) pyrrolidine-1 -carbonyl )pyrrolidin -l -carboxylate:
  • reaction mixture was diluted with DCM and washed with water, saturated NaHC0 3 solution, brine and dried over Na 2 S0 4 .
  • the solvent was evaporated and to the resulting solid, was added DCM (7 ml) and stirred for about 1 hour and filtered. The filtrate was concentrated under reduced pressure to afford the title compound (1.0 g, yield: 78.12%) as a gummy oil.
  • Step 6 Synthesis of (2S,4S)-l-(L-prolyl)-4-fluoro-2-((2-methoxyethoxy)methyl)pyrrolidine hydrochloride:
  • Step 7 Synthesis of (2S,4S)-4-fluoro-2-((2-methoxyethoxy)methyl)-l-(((S)-pyrrolidin-2-yl) methyl pyrrolidine :
  • Step 1 Synthesis of tert-butyl (S)-2-((tosyloxy)methyl)pyrrolidine-l-carboxylate:
  • Triethyl amine (0.8 ml, 5.9 mmol, 1.2 eq), followed by p-toluenesulphonyl chloride (1.04 g, 5.5 mmol, 1.1 eq) and catalytic amount of DMAP were added at 0 °C to a solution of tert-butyl (S)-2-(hydroxymethyl)pyrrolidine-l-carboxylate (Intermediate 36-step 1, 1 g, 4.9 mmol, 1.0 eq) in DCM (10 ml). The reaction was stirred for about 4 hours at room temperature. The reaction mass was diluted with water (10 ml) and the organic layer was separated. The compound was further extracted with DCM (2x10 ml).
  • Step 2 Synthesis of tert-butyl (S)-2-(pyrrolidin-l -ylmethyl)pyrrolidine-l -carboxylate:
  • Step 1 Synthesis of tert-butyl (2S,4R)-4-((methylsulfonyl)oxy)-2-(((methylsulfonyl)oxy) methyl)pyrrolidine-l-carboxylate:
  • Step 2 Synthesis of tert-butyl (3'S,5'S)-5'-(pyrrolidin-l-ylmethyl)-[l,3'-bipyrrolidine]-l '- carboxylate:
  • step 1 To a stirred solution of tert-butyl (2S,4R)-4-((methylsulfonyl)oxy)-2-(((methyl sulfonyl)oxy)methyl)pyrrolidine-l-carboxylate (step 1, 3.4 g, 9.11 mmol) in ACN (50 ml) under N 2 atmosphere, were added Cs 2 C0 3 (14.8 g, 45.5 mmol), followed by pyrrolidine (3.8 ml, 45.5 mmol) at room temperature. The reaction mixture was heated to 100 °C and continued for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to room temperature and filtered through celite and the cake was washed with DCM (twice).
  • Step 3 Synthesis of ' (3'S,5'S)-5 '-(pyrrolidin-1 -ylmethyl)-! ,3 ' -bipyrrolidine hydrochloride: To a stirred solution of tert-butyl (3'S,5'S)-5'-(pyrrolidin-l-ylmethyl)-[l,3'- bipyrrolidine]-l '-carboxylate (step 2, 1.0 g, 3.1 mmol) in dioxane (5 ml), was added 6N HCl in dioxane (10 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (0.91 g). Next reaction was carried out without any further purification.
  • Step 1 Synthesis of tert-butyl (S)-2-((4-hydroxypiperidin-l-yl)methyl)pyrrolidine-l- carboxylate:
  • Step 2 Synthesis of tert-butyl (S)-2-((4-fluoropiperidin-l-yl)methyl)pyrrolidine-l- carboxylate:
  • step 1 1-carboxylate (step 1, 1.7 g, 5.98 mmol) in DCM (25 ml) under N 2 atmosphere at 0 °C, was added DAST (1.1 ml, 6.58 mmol) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by addition ice cold water. The solution was extracted with DCM (2x50 mL) and the combined organic phases were washed with brine, dried over Na 2 S0 4 and concentrated under reduced pressure to give the crude product. Purification by column chromatography with MeOH and DCM (2:98) to afford the desired compound (0.5 g, yield: 29.4%) as an oil.
  • Step 1 Synthesis of 3 -(tert-butyl) 2-methyl (lR,2S,5S)-6,6-dimethyl-3-az bicyclo[3.1.0] hexane-2,3-dicarboxylate:
  • Step 2 Synthesis of tert-butyl (lR,2S,5S)-2-(hydroxymethyl)-6,6-dimethyl-3-azabicyclo[3.1.0] hexane-3-carboxylate:
  • Step 3 Synthesis of tert-butyl (lR,2S,5S)-6,6-dimethyl-2-(pyrrolidin-l-ylmethyl)-3-azabicyclo [3.1.0]hexane-3-carboxylate:
  • Step 4 Synthesis of (lR,2S,5S)-6,6-dimethyl-2-(pyrrolidin-l-ylmethyl)-3-az bicyclo[3.1.0] hexane dihydrochloride:
  • Step 2 Synthesis of l-((2S)-2-((lH-pyrazol-l-yl)methyl)-lX 4 -pyrrolidin-l-yl)-2,2,2-trifluoro ethan-l-one:
  • Step 1 Synthesis of tert-butyl (S)-2-(((R)-3-hydroxypyrrolidin-l-yl)methyl)pyrrolidine-l- carboxylate:
  • Step 2 Synthesis of tert-butyl (S)-2-(((R)-3-(2-methoxyethoxy)pyrrolidin-l-yl)methyl) pyrrolidine-1 -carboxylate:
  • Step 3 Synthesis of (R)-3-(2-methoxyethoxy)-l-(((S)-pyrrolidin-2-yl)methyl)pyrrolidine dihydrochloride:
  • the below intermediate 45 were prepared by the procedure similar (including reagents and reaction conditions) to the above described in the synthesis of intermediate-44 by using suitable reagents, reactents and solvents at appropriate conditions.
  • Step 1 Synthesis of tert-butyl (S)-2-((3,5-dimethyl-lH-pyrazol-l-yl)methyl)pyrrolidine-l- carboxylate:
  • Step 2 Synthesis of l-((2S)-2-((3,5-dimethyl-lH-pyrazol-l-yl)methyl)-lX 4 -pyrrolidin-l-yl)- 2,2,2-trifluoroethan-l-one:
  • Step 1 Synthesis of methyl 1 -((tert-butoxycarbonyl)amino)cyclobutane-l -carboxylate:
  • Step 2 Synthesis of tert-butyl (l-(hydrox methyl)cyclobutyl)carbamate:
  • Step 3 Synthesis of (l-((tert-butoxycarbonyl)amino)cyclobutyl)methyl 4-methylbenzene sulfonate:
  • Step 4 Synthesis of tert-butyl (1 -(pyrrolidin-1 -ylmethyl)cyclobutyl)carbamate: BocHN
  • step 3 To a stirred solution of (l-((tert-butoxycarbonyl)amino)cyclobutyl)methyl 4- methylbenzenesulfonate (step 3, 3.9 g, 10.1 mmol) in DMF (40 ml) under N 2 atmosphere, were added K 2 C0 3 (4.5 g, 33 mmol), followed by pyrrolidine (1.8 ml, 22 mmol) at room temperature. The reaction mixture was heated to 110 °C and continued for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to room temperature, filtered through celite, was washed with EtOAc (twice).
  • Step 5 Synthesis of 1 -(pyrrolidin- 1 -ylmethyljcyclobutan- 1 -amine hydro chloride :
  • step 1 tert-butyl (S)-2-(pyrrolidine-l-carbonyl)pyrrolidine-l- carboxylate (step 1, 3.5 g, 13.2 mmol, 1.0 eq) in 1,4-dioxane.HCl (30 mL) which was stirred at room temperature for about 4 hours. After completion of the reaction (monitored by TLC), the volatile solvent was concentrated to afford the desired compound as a brown liquid, which was without purification go to next step.
  • Example 1 Preparation of 2,2-dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR, 13aRJ3bR)-5a,5b,8,8 J la-pentamethyl-3a-((2-methyl-l-morpholinopropan-2-yl)carbamoyl)- l-(l-methylcvclopropyl)icosahvdro-lH-cvclo entaralchrvsen-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-3a-((2-methyl-l -morpholinopropan-2-yl)carbamoyl)-l -(1 -methylcyclopropyl) icosahydro-lH-cyclopenta[a ]chrysen-9-yl acetate:
  • reaction mixture was allowed to stir at room temperature for overnight. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with water and extracted with CH 2 C1 2 . The combined organic extracts were dried over Na 2 S0 4 , filtered and evaporated under reduced pressure. The residue was purified by column chromatography by using 8% methanol: DCM as an eluent to afford the desired product (1.4 g, yield: 95.0%) as a semi solid.
  • Step 2 Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-9-hydroxy-5a,5b,8,8, lla ⁇ entamethyl-N-(2-methyl-l-morpholinopropan-2-yl)-l-(l-methylcyclopropyl)icosahydro -3aH-cyclopenta[a]chrysene-3 -carboxamide:
  • reaction mixture was stirred at room temperature for about 48 hours. TLC indicated starting material was consumed and the desired product was observed.
  • the reaction mixture was filtered through a pad of celite and washed with CH 2 C1 2 . The filtrate was evaporated under reduced pressure and the crude was purified by column chromatography by using 1% methanol: dichloromethane as an eluent to afford the desired product (0.750 g, yield: 57.0%) as a white solid.
  • Step 3 Synthesis of 2,2-dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR) -5a,5b,8,8,lla-pentamethyl-3a-((2-methyl-l-morpholinopropan-2-yl)carbamoyl)-l-(l -methyl cyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)butanoic acid:
  • reaction mixture was heated at 90°C for overnight. TLC indicated starting material was consumed and the desired product was observed.
  • the reaction mixture was concentrated under reduced pressure, cooled to 0°C, acidified to pH ⁇ 5 with IN HC1 and extracted with CH 2 CI 2 . The combined organic extracts were washed with water, dried over Na 2 S0 4 , filtered and evaporated under reduced pressure.
  • the crude residue was purified by column chromatography by using 5% methanol: dichloromethane as an eluent to afford the desired product (0.100 g, yield: 27.0%) as a white solid.
  • Example 8 Preparation of 5-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2- ((4-ethylpiperazin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a.5b.8.8 J la-pen tamethyl-l-(l- methylcvclopropyl)icosahvdro-lH-cvclopentaralchrvsen-9-yl)oxy)-3,3-dimethyl-5-oxo pentanoic acid:
  • reaction mixture was allowed to stir at room temperature for overnight.
  • the reaction mixture was diluted with water and extracted with CH 2 CI 2 .
  • the combined organic extracts were dried over Na 2 S0 4 , filtered and evaporated under reduced pressure.
  • the residue was purified by column chromatography by using 5% methanol: DCM as an eluent gave the desired product (1.2 g, yield: 37.0%) as a light yellow solid.
  • Step 2 Synthesis of ((S)-2-((4-ethylpiperazin-l-yl)methyl)pyrrolidin-l-yl)((lR,3aS,5aR,5bR, 7aR,9S,l 1 aR,l lbR,l 3aR,l 3bR)-9-hydroxy-5a,5b, 8,8,11 a-pentamethyl-1 -(1 -methylcyclo propyl)icosahydro-3aH-cyclopenta[a]chrysen-3a-yl)methanone:
  • step 1 ((4-ethylpiperazin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b, 8,8,1 la-pentamethyl-l-(l- methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (step 1, 1.2 g, 1.74 mmol, 1.0 eq) in THF (12 mL), Methanol (12 mL) and water (12 mL) was added LiOH (0.73 g, 17.4 mmol, 10.0 eq). The reaction mixture was stirred at room temperature for about 12 hours. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was diluted with water and extracted with DCM.
  • Step 3 Synthesis of 5-(((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-3a-((S)-2-((4-ethyl piperazin-l -yl)methyl)pyrrolidine-l -carbonyl)-5a,5b,8,8,l 1 a-pentamethyl-1 -(1 -methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)-3,3-dimethyl-5-oxopentanoic acid:
  • the reaction mixture was heated at 90 °C for overnight. TLC indicated starting material was consumed and the desired product was observed.
  • the mixture was concentrated under reduced pressure, cooled to Oo C, acidified to pH ⁇ 5 with IN HC1 and extracted with CH 2 C1 2 . The combined organic extracts were washed with water, dried over Na 2 S0 4 , filtered and evaporated under reduced pressure.
  • the crude residue was purified by column chromatography by using 10% methanol and dichloromethane as an eluent gave the desired product (0.130 g, yield: 35.0%) as a white solid.
  • Example 10 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((l-((R)-3-hvdroxypyrrolidin-l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b,8,8 J la-penta methyl- l-(l-methylcvclopropyl)icosahvdro-lH-cvclopentaralchrvsen-9-yl)oxy)carbonyl)-
  • Step 1 Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-3a-((l-((R)-3-
  • reaction mixture was diluted with water and extracted with CH 2 CI 2 .
  • the combined organic extracts were dried over Na 2 S0 4 , filtered and evaporated under reduced pressure.
  • the residue was purified by column chromatography by using 4% methanol: DCM as an eluent to afford the desired product (1.0 g, yield: 71.0%) as a white solid.
  • Step 2 Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-N-(l-((R)-3- (benzyloxy)pyrrolidin-l-yl)-2-methylpropan-2-yl)-9-hydroxy-5a,5b,8,8,lla-pentamethyl-l-(l- methylcyclopropyl)icosahyd -3aH-cyclopenta[a]chrysene-3a-carboxamide:
  • reaction mixture was stirred at room temperature for about 48 hours. TLC indicated starting material was consumed and the desired product was observed.
  • the reaction mixture was filtered through a pad of celite and washed with CH 2 CI 2 . The filtrate was evaporated under reduced pressure and the crude was purified by column chromatography by using 4% methanol: dichloromethane as an eluent to afford the desired product (0.800 g, yield: 84.0%) as a white solid.
  • Step 3 Synthesis of 1-benzyl 3-((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-3a-((l- ((R)-3-(benzyloxy)pyrrolidin-l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b,8,8,lla- pentamethyl-1 -( 1 -methylcyclopropyl)icosahydro-l H-cyclopenta[a] chrysen-9-yl) (lR,3S)-2,2- dimethylcyclobutane- -dicarboxylate:
  • the reaction mixture was raised to room temperature and stirred for overnight. TLC indicated starting material was consumed and the desired product was observed.
  • the reaction mixture was evaporated under reduced pressure and diluted with water.
  • the aqueous layer was extracted with CH 2 CI 2 .
  • the organic layer was washed with water and brine solution.
  • the combined organic layers were dried over Na 2 S0 4 , filtered and evaporated under reduced pressure.
  • the crude residue was purified by column chromatography by using 3% MeOH and DCM as an eluent to obtain the desired product (0.400 g, yield: 40.0%) as a white solid.
  • Step 4 Synthesis of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-3a-((l- ((R)-3-hydroxypyrrolidin-l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b,8,8,lla-pentamethyl- 1-(1 -methylcyclopropyl)icosahydro-lH-cyclopenta[a ] chrysen-9-yl)oxy )carbonyl)-2,2- dimethylcyclobutane-l-carboxylic acid:
  • Example 55 Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((S)-2-((l J-dioxidothiomorpholino)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8 J la-penta methyl- l-(l-methylcyclopropyl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)carbonyl)-
  • reaction mixture was allowed to stir at room temperature for overnight.
  • the reaction mixture was diluted with water and extracted with CH 2 C1 2 .
  • the combined organic extracts were dried over Na 2 S0 4 , filtered and evaporated under reduced pressure.
  • the residue was purified by column chromatography by using 15% EtOAc and hexane as an eluent gave the desired product (6.0 g, yield: 85.0%) as an off white solid.
  • reaction mixture was stirred at room temperature for about 12 hours. TLC indicated starting material was consumed and the desired product was observed.
  • the reaction mixture was distilled out completely under vacuum to get crude residue.
  • the residue was purified by column chromatography by using 15% EtOAc and hexane as an eluent gave the desired product (5.50 g, yield: 98.0%) as an off white solid.
  • Step 3 Synthesis of 1 -benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8, lla-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-(thiomorpholinomethyl)pyrrolidine-l- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-dimethylcyclobutane-l,3- dicarboxylate:
  • reaction mixture was raised to room temperature stirred for overnight. TLC indicated starting material was consumed and the desired product was observed.
  • the reaction mixture was evaporated under reduced pressure and diluted with water.
  • the aqueous layer was extracted with CH 2 CI 2 .
  • the organic layer was washed with water and brine solution.
  • the combined organic layers were dried over Na 2 S0 4 , filtered and evaporated under reduced pressure.
  • the crude residue was purified by column chromatography by using 15% EtOAc and Hexane as an eluent to obtain the desired product (5.50 g, yield: 72.0%) as a white solid.
  • Step 4 Synthesis of 1 -benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-3a-((S)-2- ((1,1 -dioxidothiomorpholino )methyl )pyrrolidine-l -carbonyl)-5a,5b, 8, 8, 11 a-pentamethyl-1 - (l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-dimethylcyclo butane -1,3 -dicarboxyla
  • the reaction was stirred at 0 °C for about 1.5 hours.
  • the reaction mixture was diluted with EtOAc, washed with saturated aqueous NaHC0 3 , brine, dried over MgS0 4 , and filtered.
  • the filtrate was evaporated under reduced pressure, the crude residue was purified by column chromatography by using 3% MeOH and DCM as an eluent to obtain the desired product (0.7 g, yield: 56.0%) as a white solid.
  • Step 5 Synthesis of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-3a-((S)-2- ((1,1 -dioxidothiomorpholino )methyl )pyrrolidine-l -carbonyl)-5a,5b, 8, 8, 11 a-pentamethyl-1 - (l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl cyclo butane- 1-carboxylic acid:
  • reaction mixture was stirred in hydrogen atmosphere at room temperature for overnight. TLC indicated starting material was consumed and the desired product was observed.
  • the reaction mixture was filtered through a pad of celite and was washed with MeOH and DCM. The filtrate was evaporated under reduced pressure, the crude residue was purified by column chromatography by using 5%
  • Example 56 Synthesis of (lS,3R)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR, 13aR,13bR)-5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-(pyrrolidin-l-yl methyl)pyrrolidine-l-carbonyl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)carbonyl) cyclobutane-l-carboxylic acid:
  • Step 1 Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla-penta methyl-1 -( 1 -methylcyclopropyl)-3a-( (S)-2-(pyrrolidin-l -ylmethyl)pyrrolidine-l -carbonyl) icosahydro-lH-cyclopenta[a ]chrysen-9-yl acetate:
  • reaction mixture was allowed to stir at room temperature for overnight.
  • the reaction mixture was diluted with water and extracted with CH 2 C1 2 .
  • the combined organic extracts were dried over Na 2 S0 4 , filtered and evaporated under reduced pressure.
  • the residue was purified by column chromatography by using 4% methanol: DCM as an eluent gave the desired product (1.6 g, yield: 64.0%) as a white solid.
  • Step 2 Synthesis of ((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-9-hydroxy-5a,5b,8,8, lla-penta methyl-l-(l-methylcyclopropyl)icosahydro-3aH-cyclopenta[a]chrysen-3a-yl)((S)- 2-(pyrrolidin-l-ylmethyl)pyrrolidin-l- l methanone:
  • reaction mixture was stirred at room temperature for about 12 hours. TLC indicated starting material was consumed and the desired product was observed.
  • the reaction mixture was concentrated and this reaction mixture was diluted with water solid product was obtained which solid was filtered and dried.
  • the obtained crude was purified by column chromatography by using 5% methanol and dichloromethane as an eluent gave the desired product (1.0 g, yield: 71.0%) as a white solid.
  • Step 3 Synthesis of 1 -benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8, lla-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidine-l- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)(lS,3R)-2,2-dimethylcyclobutane-l,3- dicarboxylate:
  • Step 4 Synthesis of (lS,3R)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR, 13bR)-5a,5b,8,8,l 1 a-pentamethyl-1 -(1 -methylcyclopropyl)-3a-((S)-2-(pyrrolidin-l -ylmethyl) pyrrolidine-1 -carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane- 1 -carboxylic acid:
  • reaction mixture was stirred in hydrogen atmosphere at room temperature for overnight. TLC indicated starting material was consumed and the desired product was observed.
  • the reaction mixture was filtered through a pad of celite and was washed with MeOH and DCM. The filtrate was evaporated under reduced pressure, the crude residue was purified by column chromatography by using 8% methanol and DCM as an eluent gave the desired compound (0.210 g, yield: 34.0%) as an off white solid.
  • Example 61 Preparation of 2,2-dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR, 13aRJ3bR)-5a,5b,8,8 J la-pentamethyl-3a-((2-methyl-l-(piperidin-l-yl)propan-2-yl) carbamoyl)- l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)butanoic acid:
  • Step 1 Synthesis of (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla-penta methyl-3a-((2-methyl-l -(piperidin-l -yl)propan-2-yl)carbamoyl)-l -(prop-1 -en-2-yl) icosahydro-lH-cyclopenta[a ]chrysen-9-yl acetate:
  • reaction mixture was allowed to stir at room temperature for overnight.
  • the reaction mixture was diluted with water and extracted with CH 2 C1 2 .
  • the combined organic extracts were dried over Na 2 S0 4 , filtered and evaporated under reduced pressure.
  • the residue was purified by column chromatography by using 8% methanol: DCM as an eluent to afford the desired product (1.0 g, yield: 68.0%) as a white solid.
  • Step 2 Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-9-hydroxy-5a,5b,8,8, 11 a-pentamethyl-N -(2 -methyl- 1 -(piperidin-l -yl)propan-2-yl)-l -(prop-1 -en-2-yl)icosahydro- 3aH-cyclopenta[a]chrysene-3a-carboxamide:
  • reaction mixture was stirred at room temperature for about 48 hours. TLC indicated starting material was consumed and the desired product was observed.
  • the reaction mixture was filtered through a pad of celite and washed with CH 2 CI 2 . The filtrate was evaporated under reduced pressure and the crude was purified by column chromatography by using 1% methanol: dichloromethane as an eluent to afford the desired product (0.600 g, yield: 64.0%) as a white solid.
  • Step 3 Synthesis of 2,2-dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR, 13bR)-5a,5b,8, 8,11 a-pentamethyl- 3 a-((2 -methyl- 1 -(piperidin-l -yl)propan-2-yl)carbamoyl)-l - (prop-1 -en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)butanoic acid:
  • the reaction mixture was heated at 90°C for overnight. TLC indicated starting material was consumed and the desired product was observed.
  • the mixture was concentrated under reduced pressure, cooled to 0°C, acidified to pH ⁇ 5 with IN HC1 and extracted with CH 2 CI 2 . The combined organic extracts were washed with water, dried over Na 2 S0 4 , filtered and evaporated under reduced pressure.
  • the crude residue was purified by column chromatography by using 5% methanol: dichloromethane as an eluent to afford the desired product (0.050 g, yield: 19.0%) as a white solid.
  • Example 66 synthesis of Synthesis of 5-(lR.3aS.5aR.5bR.7aR.9S.l laR.l lbR.13aR.13bR)-
  • Step 1 Synthesis of (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-3a-(l-(4-ethylpiperazin -l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b,8,8,lla-pentamethyl-l-(prop-l-en-2-yl) icosahydro-l H-cyclopenta[a ]chrysen-9-yl acetate:
  • reaction mixture was allowed to stir at room temperature for overnight.
  • the reaction mixture was diluted with water and extracted with CH 2 C1 2 .
  • the combined organic extracts were dried over Na 2 S0 4 , filtered and evaporated under reduced pressure.
  • the residue was purified by column chromatography by using 3% methanol: DCM as an eluent gave the desired product (6.0 g, yield: 63.0%) as a solid.
  • Step 2 Synthesis of (lR,3aS,5aR,5bR, 7aR,9S,l 1 aR,l lbR,l 3aR,l 3bR)-N-(l -(4-ethylpiperazin- l-yl)-2-methylpropan-2-yl)-9-hydroxy-5a,5b,8,8,lla-pentamethyl-l-(prop-l-en-2-yl) icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide:
  • reaction mixture was stirred at room temperature for about 48 hours. TLC indicated starting material was consumed and the desired product was observed.
  • the reaction mixture was filtered through a pad of celite and washed with CH 2 CI 2 . The filtrate was evaporated under reduced pressure and the crude was purified by column chromatography by using 3% methanol: dichloromethane as an eluent gave the desired product (3.50 g, yield: 62.0%) as a white solid.
  • Step 3 Synthesis of 5-(lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-3a-(l-(4-ethyl piperazin-l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b,8,8,lla-pentamethyl-l-(prop-l-en-2- yl) icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)-3,3-dimethyl-5-oxopentanoic acid:
  • the reaction mixture was heated at 90 °C for overnight. TLC indicated starting material was consumed and the desired product was observed.
  • the mixture was concentrated under reduced pressure, cooled to 0 0 C, acidified to pH ⁇ 5 with IN HC1 and extracted with CH 2 CI 2 . The combined organic extracts were washed with water, dried over Na 2 S0 4 , filtered and evaporated under reduced pressure.
  • the crude residue was purified by column chromatography by using 4% methanol and dichloromethane as an eluent gave the desired product (0.130 g, tield: 28.0%) as a white solid.
  • Example 67 were prepared by the procedure similar to example-66 by using corresponding intermediates inpresence of suitable reagents, reactents and solvents at appropriate reaction conditions.
  • the corresponding intermediates and the charctrization data are mentioned in the below table.
  • Example 68 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-((2-methyl-l-(pyrrolidin-l-yl)propan-2-yl) carbamoyl)- l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)carbonyl) cyclobutane-l-carboxylic acid:
  • Step 1 Synthesis (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla-penta methyl-3a-((2-methyl-l -(pyrrolidin-l -yl)propan-2-yl)carbamoyl)-l -(prop-1 -en-2-yl) icosahydro-lH-cyclopenta[a ]chrysen-9-yl acetate:
  • reaction mixture was allowed to stir at room temperature for overnight.
  • the reaction mixture was diluted with water and extracted with CH 2 C1 2 .
  • the combined organic extracts were dried over Na 2 S0 4 , filtered and evaporated under reduced pressure.
  • the residue was purified by column chromatography by using 3% methanol: DCM as an eluent to afford the desired product (2.5 g, yield: 51.0%) as a white solid.
  • Step 2 Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-9-hydroxy-5a,5b,8,8, 11 a-pentamethyl-N -(2 -methyl- 1 -(pyrrolidin-l -yl)propan-2-yl)-l -(prop-1 -en-2-yl)icosahydro- 3aH-cyclopenta[a ]chrysene-3a-carboxamide:
  • reaction mixture was stirred at room temperature for about 48 hours. TLC indicated starting material was consumed and the desired product was observed.
  • the reaction mixture was filtered through a pad of celite and washed with CH 2 C1 2 . The filtrate was evaporated under reduced pressure and the crude was purified by column chromatography by using 3% methanol: dichloromethane as an eluent to afford the desired product (1.5 g, yield: 72.0%) as a white solid.
  • Step 3 Synthesis of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-5a,5b, 8,8,11 a-pentamethyl- 3 a-((2 -methyl- 1 -(pyrrolidin-l -yl)propan-2-yl)carbamoyl)-l -(prop-1 -en- 2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-dimethylcyclobutane-l,3- dicarboxylate:
  • the reaction mixture was raised to room temperature and stirred for overnight. TLC indicated starting material was consumed and the desired product was observed.
  • the reaction mixture was evaporated under reduced pressure and diluted with water.
  • the aqueous layer was extracted with CH 2 CI 2 .
  • the organic layer was washed with water and brine solution.
  • the combined organic layers were dried over Na 2 S0 4 , filtered and evaporated under reduced pressure.
  • the crude residue was purified by column chromatography by using 4% MeOH and DCM as an eluent to obtain the desired product (1.2 g, yield: 58.0%) 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-3a-((2-methyl-l-(pyrrolidin-l-yl)propan-2-yl)carbamoyl)-
  • Example 91 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-((2-(pyrrolidin-l-yl methyl)- lH-pyrrol-l-yl)methyl)pyrrolidine-l-carbonyl)icosahydro-lH-cyclopentaralchrysen- 9-yl)oxy)carbonyl)cyclobutane- 1-carboxylic acid:
  • Step 1 Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla-penta methyl-1 -(prop-1 -en-2-yl)-3a-((S)-2-((2-(pyrrolidin-l -ylmethyl)-lH-pyrrol-l-yl)methyl) pyrrolidine-1 -carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate:

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Abstract

The present invention relates to novel betulinic substituted amide compounds of formula (I); and pharmaceutically acceptable salts thereof, wherein R1, R2, R3, R4, R5, R6, R7, R8, X, Y, Z1, Z2, Z3 and are Formula (II) as defined herein. The invention novel betulinic substituted amide derivatives, related compounds, and pharmaceutical compositions useful for the therapeutic treatment of viral diseases and particularly HIV mediated diseases.

Description

NOVEL BETULINIC SUBSTITUTED AMIDE DERIVATIVES AS HIV
INHIBITORS
This application claims the benefit of Indian provisional application no 3851/CHE/2015 filed on 28th July 2015 which is hereby incorporated by reference in its entirety.
FIELD OF THE INVENTION
The present invention relates to novel betulinic substituted amide derivatives and related compounds, compositions useful for therapeutic treatment of viral diseases and particularly HIV mediated diseases.
BACKGROUND OF THE INVENTION
The Human Immunodeficiency Virus (HIV) has now been established as the causative agent of the Acquired Immunodeficiency Syndrome (AIDS) for over 20 years (Science 1983, 220,868-871; N.Eng.J.Med.1984, 311, 1292-1297). AIDS is characterized by the destruction of the immune system, particularly of CD4+T-cells. HIV is a retrovirus, and the HIV life cycle encompasses several crucial steps, starting from the attachment of the virus to the host cell membrane and finishing with the release of progeny virons from the cell.
The natural compound betulinic acid, isolated from Syzygium clavifolium and several other plant species was found to possess anti-HIV activity. Chemical modifications were undertaken by several research groups in an attempt to identify potent anti-HIV agents by making semi-synthetic analogs of betulinic acid, leading to the discovery of bevirimat as a compound with a novel mechanism of action (J. Nat. Prod. 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. 2004,78(2): 922-9; J. Biol. Chem. 2005,280(51):42149-55; J. Virol. 2006,80(12): 5716-22) and to be a first-in-class maturation inhibitor with a potent activity against HIV-1. Bevirimat went up to phase 2 clinical trials, in clinic despite optimal plasma concentrations, not all patients given bevirimat have a robust viral load reduction. It was reported that non-respondant patients had more frequent base line Gag polymorphisms near the capsid SP-1 cleavage site than responders. (HIV gag polymorphism determines treatment response to bevirimat. XVII international HIV drug resistance work shop June 10- 14, 2008, Sitges, Spain).
Encouraged by these developments, medicinal chemists started exploring betulinic acid derivatives and related compounds intensively for their therapeutic activities. For example, WO 2016001820 describes novel betulinic proline imidazole derivatives as HIV inhibitors; WO 2015198263 describes novel betulinic proline substituted derivatives as hiv inhibitors; WO 2014/105926 describes novel betulinic acid proline derivatives as HIV inhibitors; WO 2013/160810 describes novel betulinic acid derivatives as HIV inhibitors; molecular pharmaceutics, volume: 9, issue: 11, pages: 3147-3159, 2012 describes BBA, a derivative of 23-hydroxybetulinic acid, potently reverses ABCB 1 -mediated drug resistance in vitro and in vivo; European Journal of Medicinal Chemistry, Volume: 46, Issue: 6, Pages: 2490-2502, 2011 describes synthesis and antiproliferative evaluation of 23-hydroxybetulinic acid derivatives; WO 2011/007230 describes lupeol-type triterpene derivatives as antivirals; WO 2008/115281 describes preparation of triterpene derivatives for therapeutic use in the treatment of viral infections; WO 2007/002411 describes preparation of triterpenes as antiviral agents; WO 2006/053255 describes novel Betulin derivatives, Preparation thereof & use thereof; WO 2002091858 describes anxiolytic marcgraviaceae compositions containing betulinic acid, betulinic acid derivatives, and methods; CN 102108092 describes 23- Hydroxybetulinic acid derivative, its preparation method and application as antitumor agents and tumor multidrug resistance reversing agents; Bioorganic & Medicinal Chemistry Letters (2008), 18(24), 6377-6380 describes triterpene based compounds with potent anti-maturation activity against HIV-1; Bioorganic & Medicinal Chemistry Letters 9 (1999) 1201-1204 describes preparation of amino acid conjugates of betulinic acid with activity against human melanoma; Pharmazie, Volume: 53, Issue: 10, Pages: 677-680, Journal, 1998 describes synthesis of amino acid conjugates of 3a-hydroxylup-20(29)-ene-23,28-dioic acid; WO 2011/153319 describes C-28 amides of modified C-3 betulinic acid derivatives as HIV maturation inhibitors; WO 2011/153315 describes modified C-3 betulinic acid derivatives as HIV maturation inhibitors; WO 2009/082819 describes novel lupane derivatives; WO 2009/100532 discloses novel 17 β lupine derivatives as anti-HIV agents; The patent publication WO 2014/093941 Al describers pharmaceutical compositions of betulin derivatives; WO 2008/057420 extended triterpene derivatives as antiretroviral agents; US 2013/0096094 describes preparation of derivatives of 3-0-(3',3'-dimethylsuccinyl)-betulinic acid for the treatment of HIV; WO 2010/132334 describes 3,28-Disubstituted betulinic acid derivatives as Anti-HIV agents; US 2004/0204389 describes anti-HIV agents with dual sites of action.
Some additional references disclose betulinic acid related compounds. For example,
WO 2013/148067 describes preparation of 3,28-disubstituted betulinic acid derivatives as anti-HIV agents; WO 2012/154554 describes fatty acid triterpene derivatives and their uses; WO 2010/032123 describes preparation of triterpenoid compounds for pharmaceutical use; WO 2008/138200 describes preparation of lupane derivatives as NMDA and MC receptor antagonists exhibiting neuroprotective and memory enhancing activities; WO 2008/127364 describes preparation of betulinic acid derivatives for use in antiviral and anticancer pharmaceutical compositions; WO 2008/097341 describes antiviral compounds and use thereof; WO 94/26725 describes preparation of lupane antiviral agents; WO 2006/053255 describes novel Betulin derivatives, preparation thereof & use thereof; CN 103342729 describes caffeoyl-substituted pentacyclic triterpene derivative useful in treatment of various diseases and its preparation; CN 103127135 describes triterpene derivatives, their preparation method and use; US 2011/0152229 describes betulinic acid derivatives as anti-HIV agents; CN 1861627 describes synthesis of 23-hydroxybetulinic acid derivatives and application as antitumor agents; EP 542622 describes preparation of [lup-20(29)-en-28- oylamino]alkanoates as antiviral agents; CN 101648988 describes preparation of pentacyclic triterpene-28-carboxylic amide derivative containing isoxazole ring and medical application; WO 2007/141392 describes compositions comprising betulonic acid; WO 2007/141391 describes betulin derived compounds useful as antiprotozoal agents; Bioorganic & Medicinal Chemistry (2006), 14(7), 2279-2289 describes synthesis and anti-HIV activity of bi- functional betulinic acid derivatives.
Given the fact of the world wide epidemic level of AIDS, there is a strong continued need for new effective drugs for treatment of HIV infected patients, disease conditions and/or disorders mediated by HIV by discovering new compounds with novel structures and/or mechanism of action(s).
SUMMARY OF THE INVENTION
The present invention relates to the com ounds of the formula (I):
Figure imgf000004_0001
wherein, n be substituted or unsubstituted alkyl,
Figure imgf000004_0002
Figure imgf000004_0003
Figure imgf000005_0001
(wherein Rb can be hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl);
R2 can be hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted aminoacids, substituted or unsubstituted alkoxy or substituted or unsubstituted cycloalkyl;
X can be absent, O, S, CH2 or NRa (wherein Ra can be H, C(0)Rc, C(S)RC, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, or Ra is with their adjacent N and Carbon together form N-contained heterocycle (Preferably, pyrrolidine, piperdine, piperzine, or morpholine));
Y can be C(O), C(S) or CReRf (wherein Re and Rf are independently selected from H, substituted or unsubstituted alkyl);
R3 and R4 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 R3 and R4 are taken together with the carbon atoms to which they are attached to form a bond or R3 and R4 are taken together with the carbon atoms to which they are attached to form cycloalkyl or R3 and R4 are taken together with the carbon atoms to which they are attached to form epoxide;
R5 and Rc can be independently selected from H, C02Rd (wherein Rd is H, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl), or substituted or unsubstituted alkyl;
R6 and R8 can be independently selected from H, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl;
R7 can be substituted or unsubstituted alkyl or substituted or unsubstituted cycloalkyl;
R6 and R7 can be taken together with the nitrogen and carbon to which they are attached to form substituted or unsubstituted 4-7 membered heterocyclyl, substituted or unsubstituted 4-7 membered bridged heterocyclyl wherein the substituents are alkyl, heterocyclyl or -O- heterocyclyl;
R7 and R8 are taken together with the carbon atom to which they are attached to form 3-
7 membered cycloalkyl or heterocyclyl;
N-~"-\
'·-----'' can be 4-10 membered heterocyclyl or 4-10 membered heteroaryl;
Zi, Z2 and Z3 can be independently selected from H, hydroxy, halo, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkylsulfonyl, substituted or unsubstituted alkoxy, substituted or unsubstituted alkoxylalkoxy substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, substituted or unsubstituted heterocyclylalkyl, substituted or unsubstituted aminoalkyl, substituted or unsubstituted hydroxyalkyl, -O-heterocyclyl, substituted or unsubstituted heteroaryl; wherein the substituents are aryl, alkyl, alkoxy or alkoxylalkoxy including pharmaceutically acceptable salts, pharmaceutically acceptable solvates, pharmaceutically acceptable hydrates, tautomers, stereoisomers, ester prodrugs, or combination thereof.
It should be understood that the formula (I) structurally encompasses all stereoisomers, including enantiomers, diastereomers, racemates, and combinations thereof which may be contemplated from the chemical structure of the genus described herein.
It should be understood that the formula (I) structurally encompasses all tautomers.
Also contemplated are prodrugs of the compounds of the formula (I), including ester prodrugs.
According to one embodiment, there is provided a compound of formula (I), wherein
Figure imgf000006_0001
According to other embodiment, there is provided a compound of formula (I), wherein 'X' is absent.
According to yet another embodiment, there is provided a compound of formula (I), wherein R2 is hydrogen.
According to yet another embodiment, there is provided a compound of formula (I), when "X is absent, and R2 and R5 are hydrogen"; then R3 and R4 are taken together with carbon atoms to which they are attached form a bond.
According to yet another embodiment, there is provided a compound of formula (I), when "X is absent, and R2 and R5 are hydrogen"; then R3 and R4 are taken together with carbon atoms to which they are attached form cycloalkyl (preferably cyclopropyl), According to yet another embodiment, there is provided a compound of formula (I), wherein R3, R4 and R5 are hydrogen when 'X' is absent and R2 is hydrogen.
According to yet another embodiment, there is provided a compound of formula (I), wherein R6 is hydrogen.
According to yet another embodiment, there is provided a compound of formula (I), wherein R7 is alkyl (preferably methyl, isobutyl).
According to yet another embodiment, there is provided a compound of formula (I), wherein R8 is hydrogen and alkyl (preferably methyl).
According to yet another embodiment, there is provided a compound of formula (I), wherein R6 and R7 are taken together with the nitrogen and carbon atoms to which they are attached to form a substituted or unsubstituted 4-10 membered heterocyclyl.
According to yet another embodiment, there is provided a compound of formula (I), wherein the above said heteroc clyl is substituted or unsubstituted pyrrolidine; wherin the
substituents are pyrrolidine and
Figure imgf000007_0001
According to yet another embodiment, there is provided a compound of formula (I), Ύ is CReRf or -C(O)-; wherein Re and Rf are hydrogen.
According to yet another embodiment, there is provided a compound of formula (I),
N "
/ ;
is 4-10 membered heterocyclyl.
According to the preceding embodiment the said 4-10 membered heterocyclyl is morpholinyl, pyrrolidinyl, piperidinyl, piperizinyl, azepane and 8-azabicyclo[3.2.1]octane, thiomorpholine 1,1 -dioxide and thiomorpholinyl.
According to yet another embodiment, there is provided a compound of formula (I),
N ~
/ ;
'·-·---' is 4-10 membered heteroaryl.
According to the preceding embodiment the said 4-10 membered heteroaryl is pyrrole, imidazoylyl, pyrazolyl and lH-pyrrolo[2,3-b]pyridine.
Accordingly, another aspect of the present invention provides compounds of formula
(IA): Formula (IA) wherein,
Ri, R2, R3, R4, R5, R7, Rg, 'X', Re, Rf, Zi, Z2 and Z3 as defined above including pharmaceutically acceptable salts, pharmaceutically acceptable solvates, pharmaceutically acceptable hydrates, tautomers, stereoisomers, ester prodrugs, or combination thereof.
It should be understood that the formula (IA) structurally encompasses all stereoisomers, including enantiomers, diastereomers, racemates, and combinations thereof which may be contemplated from the chemical structure of the genus described herein.
It should be understood that the formula (IA) structurally encompasses all tautomers.
Also contemplated are prodrugs of the compounds of the formula (IA), including ester prodrugs.
Accordingly, yet another aspect of the present invention provides compounds of formula (IB):
Figure imgf000008_0001
wherein
Ri, R2, R3, R4, R5, R8, 'X', Ύ, Zi, Z2 and Z3 are as defined above including pharmaceutically acceptable salts, pharmaceutically acceptable solvates, pharmaceutically acceptable hydrates, tautomers, stereoisomers, ester prodrugs, or combination thereof.
It should be understood that the formula (IB) structurally encompasses all stereoisomers, including enantiomers, diastereomers, racemates, and combinations thereof which may be contemplated from the chemical structure of the genus described herein.
It should be understood that the formula (IB) structurally encompasses all tautomers. Also contemplated are prodrugs of the compounds of the formula (IB), including ester prodrugs.
In a further embodiment, there is provided a method for treating mammals infected with a virus, especially wherein said virus is HIV, comprising administering to said mammal an antiviral effective amount of a compound which is selected from the group of compounds of formula (I), and one or more pharmaceutically acceptable carriers, excipients or diluents. Optionally, the compound of formula (I) can be administered in combination with an antiviral effective amount of another AIDS treatment agent selected from the group consisting of: (a) an AIDS antiviral agent or (b) an anti-infective agent
Another embodiment of the present invention is a pharmaceutical composition comprising one or more compounds of formula (I), and one or more pharmaceutically acceptable carriers, excipients, and/or diluents; and optionally in combination with another AIDS treatment agent selected from the group consisting of: AIDS antiviral agent or anti- infective agent
In another embodiment of the invention there is provided one or more methods for preparation of the compounds of formula (I).
In one further embodiment, the present invention also encompasses the method(s) of preparation of intermediates used in the preparation of compound of formula (I).
Below are the representative examples according to formula (I) , which are illustrative in nature only and are not intended to limit to the scope of the invention (Nomenclature has been generated from ChemBioDraw Ultra 13.0 version):
2,2-dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8, 11 a-pentamethyl-3 a-((2-methyl- 1 -morpholinopropan-2-yl)carbamoyl) - 1 -( 1 -methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)butanoic acid (Example 1),
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-((2-methyl-l-(piperidin-l-yl)propan-2-yl)carbamoyl)-l-(l-methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)butanoic acid (Example 2),
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-((2-methyl-l-(pyrrolidin-l-yl)propan-2-yl)carbamoyl)-l-(l-methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)butanoic acid (Example 3),
4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-((lR,5S)-3-(3-isopropyl- 5-methyl-4H-l,2,4-triazol-4-yl)-8-azabicyclo[3.2.1]octan-8-yl)-2-methylpropan-2-yl) carbamoyl)-5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a] chrysen-9-yl)oxy)-2,2-dimethyl-4-oxobutanoic acid (Example 4), 2,2-dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8, 1 la-pentamethyl- l-(l-methylcyclopropyl)-3a-((S)-2-(pyrrolidin- l-ylmethyl)pyrrolidine- 1- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)butanoic acid (Example 5),
2,2-dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8,l la- pentamethyl- 1 - ( 1 -methylcyclopropyl) - 3a-((S)-2- (pyrrolidine- 1 -c arbonyl)pyrrolidine- 1 - carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)butanoic acid (Example 6),
4- (((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-ethylpiperazin-l- yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl- 1-(1 -methylcyclopropyl) icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)-2,2-dimethyl-4-oxobutanoic acid (Exampl
7) ,
5- (((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-ethylpiperazin-l- yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl- 1-(1 -methylcyclopropyl) icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)-3,3-dimethyl-5-oxopentanoic acid (Exampl
8) ,
5-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-ethylpiperidin-l- yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl- 1-(1 -methylcyclopropyl) icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)-3,3-dimethyl-5-oxopentanoic acid (Exampl
9) ,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-((R)-3- hydroxypyrrolidin- l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b, 8, 8,1 la-pentamethyl- 1-(1- methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl cyclobutane-l-carboxylic acid (Example 10),
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,l la-pentamethyl-3a-((2-methyl-l-morpholinopropan-2-yl)carbamoyl)-l-(l-methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid (Example 11),
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,l la-pentamethyl-3a-((2-methyl-l-(pyrrolidin-l-yl)propan-2-yl)carbamoyl)-l-(l-methyl cyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l- carboxylic acid (Example 12),
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,l la-pentamethyl-3a-((2-methyl-l-(piperidin-l-yl)propan-2-yl)carbamoyl)-l-(l-methyl cyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l- carboxylic acid (Example 13), (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-((lR,5S)-3-(3- isopropyl-5-methyl-4H-l,2,4-triazol-4-yl)-8-azabicyclo[3.2.1]octan-8-yl)-2-methylpropan-2- yl)carbamoyl)-5a,5b,8,8, 1 la-pentamethyl- l-(l-methylcyclopropyl)icosahydro- lH-cyclopenta [a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l-carboxylic acid (Example 14),
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,1 la-pentamethyl- 1-(1 -methylcyclopropyl)-3a-((S)-2-(pyrrolidin- l-ylmethyl)pyrrolidine- l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l- carboxylic acid (Example 15),
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,l la-pentamethyl-3a-((2-methyl-l-((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidin-l-yl)propan-2- yl)carbamoyl)-l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy) carbonyl)cyclobutane-l-carboxylic acid (Example 16),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(4-ethyl piperazin-l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b,8,8,l la-pentamethyl- l-(l-methyl cyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl cyclobutane-l-carboxylic acid (Example 17),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-ethyl piperazin- 1 -yl)methyl)pyrrolidine- 1 -carbonyl)-5a,5b,8,8, 11 a-pentamethyl- 1 -( 1 -methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid (Example 18),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-ethyl piperidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl- 1-(1 -methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid (Example 19),
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,1 la-pentamethyl- 1-(1 -methylcyclopropyl)-3a-((S)-2-(piperidin- l-ylmethyl)pyrrolidine- 1- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid (Example 20),
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,1 la-pentamethyl- 1-(1 -methylcyclopropyl)-3a-((R)-2-(pyrrolidin- 1 -ylmethyl)pyrrolidine- l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l- carboxylic acid (Example 21),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(azepan-l- ylmethyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl- l-(l-methylcyclopropyl) icosa ydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid (Example 22),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((lH- imidazol- l-yl)methyl)pyrrolidine- l-carbonyl)-5a,5b,8,8, 11 a-pentamethyl- 1-(1 -methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid (Example 23),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((2- isopropyl-lH-imidazol-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b, 8,8,11 a-pentamethyl- 1-(1- methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl cyclobutane-l-carboxylic acid (Example 24),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(((S)-3- (dimethylamino)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl- l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2- dimethylcyclobutane-l-carboxylic acid (Example 25),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((3,5- dimethyl- lH-pyrazol- l-yl)methyl)pyrrolidine- l-carbonyl)-5a,5b, 8, 8, 11 a-pentamethyl- 1-(1- methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl cyclobutane-l-carboxylic acid (Example 26),
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8, 11 a-pentamethyl- l-(l-methylcyclopropyl)-3a-((S)-2-(pyrrolidine- l-carbonyl)pyrrolidine- 1- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid (Example 27),
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,11 a-pentamethyl- 1 - ( 1 -methylcycloprop yl) - 3 a- ((S ) -2- (morpholinomethyl)pyrrolidine- 1 - carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid (Example 28),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4- (hydroxymethyl)piperidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b, 8,8,11 a-pentamethyl- 1 - (l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl cyclobutane-l-carboxylic acid (Example 29),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-butyl piperazin- 1 -yl)methyl)pyrrolidine- 1 -carbonyl)-5a,5b,8,8, 11 a-pentamethyl- 1 -( 1 -methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid (Example 30), (lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b,
8,8,l la-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-((4-methylpiperidin-l-yl)methyl) pyrrolidine- l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane- 1-carboxylic acid (Example 31),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4- isopropylpiperazin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b, 8,8,11 a-pentamethyl- 1-(1- methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl eye lobutane- 1-carboxylic acid (Example 32),
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,1 la-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-(((S)-2-(5-phenyl-lH-imidazol-2- yl)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) oxy)carbonyl)cyclobutane- 1-carboxylic acid (Example 33),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((lH- pyrrolo[2,3-b]pyridin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b, 8,8,11 a-pentamethyl- 1-(1- methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl eye lobutane- 1-carboxylic acid (Example 34),
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,l la-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-(((R)-2-methylpyrrolidin-l-yl) methyl)pyrrolidine-l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl) eye lobutane- 1-carboxylic acid (Example 35),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4,4- difluoropiperidin- 1 -yl)methyl)pyrrolidine- 1 -carbonyl)-5a,5b, 8,8, 11 a-pentamethyl- 1 -( 1 - methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl eye lobutane- 1-carboxylic acid (Example 36),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((3,3- difluoropyrrolidin- 1 - yl)methyl)pyrrolidine- 1 -c arbonyl) -5 a, 5b , 8 , 8 , 11 a-pentamethyl- 1 - ( 1 - methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl eye lobutane- 1-carboxylic acid (Example 37),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-fluoro piperidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid (Example 38),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(((S)-3- fluoropyrrolidin- 1 - yl)methyl)pyrrolidine- 1 -c arbonyl) -5 a, 5b , 8 , 8 , 11 a-pentamethyl- 1 - ( 1 -methyl cyclopropyl)icosa ydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclo butane- 1-carboxylic acid (Example 39),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(((S)-2-(2- hydroxypropan-2-yl)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-penta methyl- l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)- 2,2-dimethylcyclobutane- 1-carboxylic acid (Example 40),
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,1 la-pentamethyl-l-(l-methylcyclopropyl)-3a-((2S,4R)-2-(pyrrolidin-l-ylmethyl)-4-(((R)- tetrahydrofuran-3-yl)oxy)pyrrolidine-l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) oxy)carbonyl)cyclobutane- 1-carboxylic acid (Example 41),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(((S)-2- (hydroxymethyl)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl- l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2- dimethylcyclobutane- 1-carboxylic acid (Example 42),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(((S)-3-(2- methoxyethoxy)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl- 1- (l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl eye lobutane- 1-carboxylic acid (Example 43),
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,1 la-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-(((R)-3-(((R)-tetrahydrofuran-3-yl) oxy)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9- yl)oxy)carbonyl)cyclobutane- 1-carboxylic acid (Example 44),
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,1 la-pentamethyl-l-(l-methylcyclopropyl)-3a-((3'S,5'S)-5'-(pyrrolidin-l-ylmethyl)-[l,3'- bipyrrolidine]- -carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl) eye lobutane- 1-carboxylic acid (Example 45),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-cyclo hexylpiperazin- 1 - yl)methyl)pyrrolidine- 1 -c arbonyl) - 5a,5b,8,8,l l a-pentamethyl- 1 - ( 1 -methyl cyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclo butane- 1-carboxylic acid (Example 46),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-(2- methoxyethyl)piperazin- l-yl)methyl)pyrrolidine- l-carbonyl)-5a,5b, 8, 8,1 la-pentamethyl- 1- (l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl eye lobutane- 1-carboxylic acid (Example 47), (lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b,
8,8,l la-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-((4-methylpiperazin-l-yl)methyl) pyrrolidine- l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane- 1-carboxylic acid (Example 48),
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,11 a-pentamethyl- 1 - ( 1 -methylcyclopropyl) - 3a-((S)-2- (thiomorpholinomethyl)pyrrolidine- l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l- carboxylic acid (Example 49),
(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-((2-methyl-lH-imidazol-l-yl)methyl)pyrrolidine-l-carbonyl)- l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)
eye lobutane- 1-carboxylic acid (Example 50),
(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-lH-imidazol-l-yl)methyl)pyrrolidine-l-carbonyl)- l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)
eye lobutane- 1-carboxylic acid (Example 51),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((lR,2S,5S)-6,6- dimethyl-2-(pyrrolidin-l-ylmethyl)-3-azabicyclo[3.1.0]hexane-3-carbonyl)-5a,5b, 8,8,1 la- pentamethyl-l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane- 1-carboxylic acid (Example 52),
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,l la-pentamethyl-3a-((2-methyl-l-(4-(methylsulfonyl)piperazin-l-yl)propan-2-yl) carbamoyl)- l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl) eye lobutane- 1-carboxylic acid (Example 53),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(((2S,4S)-4- fluoro-2-((2-methoxyethoxy)methyl)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b, 8,8,l la-pentamethyl-l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) oxy)carbonyl)-2,2-dimethylcyclobutane- 1-carboxylic acid (Example 54),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((l,l- dioxidothiomorpholino)methyl)pyrrolidine-l-carbonyl)-5a,5b, 8,8,11 a-pentamethyl- 1-(1- methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl eye lobutane- 1-carboxylic acid (Example 55),
(lS,3R)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,l la-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidine- l-carbonyl)icosa ydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l- carboxylic acid (Example 56),
(lS,3R)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-ethyl piperazin- 1 -yl)methyl)pyrrolidine- 1 -carbonyl)-5a,5b, 8,8, 11 a-pentamethyl- 1 -( 1 -methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid (Example 57),
(lS,3R)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-fluoro piperidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid (Example 58),
(lS,3R)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,l la-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-(morpholinomethyl)pyrrolidine-l- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid (Example 59),
(lS,3R)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,l la-pentamethyl-l-(l-methylcyclopropyl)-3a-((R)-2-(pyrrolidin-l-ylmethyl)pyrrolidine-
1- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l- carboxylic acid (Example 60),
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-((2-methyl-l-(piperidin-l-yl)propan-2-yl)carbamoyl)-l-(prop-l-en-2-yl) icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)butanoic acid (Example 61),
4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(4-ethylpiperazin-l-yl)-
2- methylpropan-2-yl)carbamoyl)-5a,5b,8,8,l la-pentamethyl-l-(prop-l-en-2-yl)icosahydro- lH-cyclopenta[a]chrysen-9-yl)oxy)-2,2-dimethyl-4-oxobutanoic acid (Example 62),
2,2-dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8,l la- pentamethyl-l-(prop-l-en-2-yl)-3a-((S)-2-(pyrrolidine-l-carbonyl)pyrrolidine-l-carbonyl) icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)butanoic acid (Example 63),
4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(((R)-3-(2-methoxy ethoxy)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl-l-(prop-l- en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)-2,2-dimethyl-4-oxobutanoic acid (Example 64),
4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-ethylpiperazin-l- yl)methyl)pyrrolidine-l-carbonyl)-5a,5b, 8,8,11 a-pentamethyl- 1 -(prop- l-en-2-yl)icosahydro- lH-cyclopenta[a]chrysen-9-yl)oxy)-2,2-dimethyl-4-oxobutanoic acid (Example 65), 5-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(4-ethylpiperazin-l-yl)-
2-methylpropan-2-yl)carbamoyl)-5a,5b,8,8,l la-pentamethyl- l-(prop-l-en-2-yl)icosahydro- lH-cyclopenta[a]chrysen-9-yl)oxy)-3,3-dimethyl-5-oxopentanoic acid (Example 66),
5-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-ethylpiperazin-l- yl)methyl)pyrrolidine- l-carbonyl)-5a,5b, 8, 8,1 la-pentamethyl- l-(prop-l-en-2-yl)icosahydro- lH-cyclopenta[a]chrysen-9-yl)oxy)-3,3-dimethyl-5-oxopentanoic acid (Example 67),
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8, 1 la-pentamethyl-3a-((2-methyl-l-(pyrrolidin- l-yl)propan-2-yl)carbamoyl)- l-(prop- 1-en- 2-yl)icosahydro- lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane- 1-carboxylic acid (Example 68),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(4-ethyl piperazin-l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b, 8, 8,1 la-pentamethyl- l-(prop-l -en-2- yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid (Example 69),
(lR,3R)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,l la-pentamethyl-3a-((2-methyl-l-(piperidin-l-yl)propan-2-yl)carbamoyl)-l-(prop-l-en- 2-yl)icosahydro- lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane- 1-carboxylic acid (Example 70),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(4-benzyl piperazin-l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b, 8, 8,1 la-pentamethyl- l-(prop-l -en-2- yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid (Example 71),
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8, 1 la-pentamethyl- l-(prop- l-en-2-yl)-3a-((S)-2-(pyrrolidine- l-carbonyl)pyrrolidine- 1- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane- 1-carboxylic acid (Example 72),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-((R)-3-(2- methoxyethoxy)pyrrolidin- l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b,8,8, 1 la-pentamethyl -l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl eye lobutane- 1-carboxylic acid (Example 73),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(4-(2-methoxy ethoxy)piperidin- l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b, 8, 8,1 la-pentamethyl- l-(prop- l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane- 1-carboxylic acid (Example 74), (lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b,
8,8,11 a-pentamethyl- 1 -(prop- 1 -en-2-yl)-3a-(( 1 -(pyrrolidin- 1 -ylmethyl)cyclobutyl) carbamoyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid (Example 75),
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,11 a-pentamethyl- 1 -(prop- 1 -en-2-yl)-3a-(( 1 -(pyrrolidin- 1 -ylmethyl)cyclopentyl) carbamoyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid (Example 76),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(4-ethoxy-4- phenylpiperidin-l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b, 8,8,11 a-pentamethyl- l-(prop- l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane- 1-carboxylic acid (Example 77),
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8, 11 a-pentamethyl- l-(prop- l-en-2-yl)-3a-(((S)- 1 -(pyrrolidin- l-yl)propan-2-yl)carbamoyl) icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid (Example 78),
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8, 1 la-pentamethyl-3a-(((S)-4-methyl- l-(piperidin- l-yl)pentan-2-yl)carbamoyl)-l-(prop- 1- en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid (Example 79),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-ethyl piperazin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b, 8,8,11 a-pentamethyl- l-(prop-l -en-2- yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid (Example 80),
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8, 1 la-pentamethyl-3a-((S)-2-(piperidin- l-ylmethyl)pyrrolidine-l-carbonyl)- l-(prop- l-en-2- yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid (Example 81),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-ethyl piperidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl-l-(prop-l-en-2-yl) icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid (Example 82),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((3,5- dimethyl- lH-pyrazol- l-yl)methyl)pyrrolidine- l-carbonyl)-5a,5b, 8, 8, 11 a-pentamethyl- 1- (prop-l-en-2-yl)icosa ydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclo butane- 1-carboxylic acid (Example 83),
(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-(morpholinomethyl)pyrrolidine-l-carbonyl)-l-(prop-l-en-2- yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane- 1-carboxylic acid (Example 84),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((lH- pyrazol-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl-l-(prop-l-en-2-yl) icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid (Example 85),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(((R)-3-(2- methoxyethoxy)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl-l- (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclo butane- 1-carboxylic acid (Example 86),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(((R)-3-(2- methoxyethoxy)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl-l- (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclo butane- 1-carboxylic acid (Example 87),
(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-((R)-2-(pyrrolidin-l-ylmethyl)pyrrolidine-l- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane- 1-carboxylic acid (Example 88),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(((S)-2-(2- hydroxypropan-2-yl)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-penta methyl- l-(prop-l -en-2-yl)icosahydro- lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2- dimethylcyclobutane- 1-carboxylic acid (Example 89),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((lR,2S,5S)-6,6- dimethyl-2-(pyrrolidin-l-ylmethyl)-3-azabicyclo[3.1.0]hexane-3-carbonyl)-5a,5b,8,8,l la- pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2- dimethylcyclobutane- 1-carboxylic acid (Example 90),
(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-((2-(pyrrolidin-l-ylmethyl)-lH-pyrrol-l- yl)methyl)pyrrolidine-l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl) eye lobutane- 1-carboxylic acid (Example 91), (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-((2-methyl-lH-pyrrol-l-yl)methyl)pyrrolidine-l-carbonyl)-l- (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l- carboxylic acid (Example 92),
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8, 1 la-pentamethyl- l-(prop- l-en-2-yl)-3a-((S)-2-(pyrrolidin- l-ylmethyl)pyrrolidine- 1- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid (Example 93),
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((lH- pyrrol-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl- l-(prop-l-en-2-yl) icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid (Example 94),
(lS,3R)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8, 1 la-pentamethyl- l-(prop- l-en-2-yl)-3a-((S)-2-(pyrrolidin- l-ylmethyl)pyrrolidine- 1- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid (Example 95),
(lR,3S)-3-((((lS,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-l-isopropyl-5a,5b, 8,8,l la-pentamethyl-3a-((2-methyl-l-(pyrrolidin-l-yl)propan-2-yl)carbamoyl)icosahydro- lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l-carboxylic acid (Example 96),
(lR,3S)-3-((((lS,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-l-isopropyl-5a,5b,8,8, l la-pentamethyl-3a-((l-(pyrrolidin-l-ylmethyl)cyclobutyl)carbamoyl)icosahydro-lH-cyclo penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane- 1-carboxylic acid (Example 97),
(lR,3S)-3-((((lS,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-l-isopropyl-3a-((l- ((R)-3-(2-methoxyethoxy)pyrrolidin- l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b,8,8, 1 la- pentamethylicosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclo butane- 1-carboxylic acid (Example 98),
(lR,3S)-3-((((lS,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-l-isopropyl-3a-((l-(4- (2-methoxyethoxy)piperidin- 1 -yl)-2-methylpropan-2-yl)carbamoyl)-5 a,5b, 8 ,8,11 a-penta methylicosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid (Example 99),
(lR,3S)-3-((((lS,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-l-isopropyl-5a,5b,8,8, l la-pentamethyl-3a-((l-(pyrrolidin-l-ylmethyl)cyclopentyl)carbamoyl)icosahydro-lH-cyclo penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane- 1-carboxylic acid (Example
100),
(lR,3S)-3-((((lS,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-l-isopropyl-5a,5b,8,8, l la-pentamethyl-3a-(((S)-4-methyl-l-(pyrrolidin-l-yl)pentan-2-yl)carbamoyl)icosahydro- lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane- 1-carboxylic acid (Example 101),
(lR,3S)-3-((((lS,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(((S)-2-(2- hydroxypropan-2-yl)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)-l-isopropyl-5a,5b,8,8, l la-pentamethylicosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclo butane- 1-carboxylic acid (Example 102), and
(lR,3S)-3-((((lS,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-l-isopropyl-5a,5b,8,8, l la-pentamethyl-3a-((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidine-l-carbonyl)icosahydro-lH- cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane- 1-carboxylic acid (Example 103); or pharmaceutically acceptable salts, solvates, including hydrates and prodrugs of compounds are also contemplated.
The present invention also provides a pharmaceutical composition that includes at least one compound as described herein and at least one pharmaceutically acceptable excipient (such as a pharmaceutically acceptable carrier or diluent). Specifically, the pharmaceutical composition comprises a therapeutically effective amount of at least one compound described herein. The compound(s) present in the composition may be associated with a pharmaceutically acceptable excipient (such as a carrier or a diluent) or may be diluted by a carrier, or enclosed within a carrier which may be in the form of a capsule, sachet, or other container.
The compounds and pharmaceutical compositions described herein are useful in the treatment of diseases, conditions and/or disorders mediated by viral infections.
The present invention further provides a method of treating a disease, condition and/or disorder mediated by viral infections in a subject in need thereof by administering to the subject one or more compounds described herein in a therapeutically effective amount to cause that infection, specifically in the form of a pharmaceutical composition.
Also provided herein are processes for preparing compounds described herein.
The invention provides a method for preventing; ameliorating or treating a HIV mediated disease, disorder or syndrome in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a compound of the invention. The invention further provides a method, wherein the HIV mediated disease, disorder or syndrome is like AIDS, AIDS related complex, or a syndrome characterized by symptoms such as persistent generalized lymphadenopathy, fever and weight loss, or a retroviral infection genetically related to AIDS.
Anti HIV inhibitory potential of the compounds of present invention may be demonstrated by any one or more methodologies known in the art, such as by using the assays described in Mossman T, December 1983, Journal of immunological methods, 65 (1- 2), 55-63 and SPC Cole, cancer chemotherapy and Pharmacology, 1986, 17, 259-263.
DETAILED DESCRIPTION OF THE INVENTION
The present invention provides betulinic substituted amide derivatives and related compounds, which may be used as antiviral particularly as anti-HIV compounds and processes for the synthesis of these compounds. Pharmaceutically acceptable salts, pharmaceutically acceptable solvates, enantiomers, diastereomers of the derivatives, together with pharmaceutically acceptable carriers, excipients or diluents, which can be used for the treatment of diseases, condition and/or disorders mediated by viral infections, are also provided.
The following definitions apply to the terms as used herein:
The terms "halogen" or "halo" includes fluorine, chlorine, bromine, or iodine.
The term "alkyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation, having from one to eight carbon atoms, and which is attached to the rest of the molecule by a single bond, e.g., methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), isobutyl, n-butyl, n-pentyl, and 1,1-dimethylethyl (t-butyl).
The term "alkoxy" refers to a straight or branched hydrocarbon chain with oxygen radical consisting carbon and hydrogen atoms, containing saturation or unsaturation, having from one to eight carbon atoms, and which is attached through oxygen atom to the rest of the molecule by a single bond, e.g., methyloxy, ethyloxy, n-propyloxy, 1-methylethyloxy (isopropyloxy), n-butyloxy, n-pentyloxy, and 1,1-dimethylethyloxy (t-butyloxy).
The term "alkoxylalkoxy" refers to a straight or branched hydrocarbon chain with oxygen radical consisting carbon atom, hydrogen atom and alkoxy groups, containing saturation or unsaturation, having from one to eight carbon atoms, and which is attached through oxygen atom to the rest of the molecule by a single bond, e.g., 2- (methyloxy)ethyloxy, 2-(ethyloxy)ethyloxy, 2-(n-propyloxy)ethyloxy, and 3- (isopropyloxy)butyloxy.
The term "cycloalkyl" denotes a non-aromatic mono or multicyclic ring system of from 3 to about 12 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Examples of multicyclic cycloalkyl groups include, but are not limited to, perhydronapththyl, adamantyl and norbornyl groups, bridged cyclic groups and spirobicyclic groups, e.g., spiro (4,4) non-2-yl.
The term "alkyl sulfonyl" refers to a group designates an "alkyl-S02-" group, wherein alkyl is as defined above. Examples of preferred alkyl sulfonyl groups of the invention include methyl sulfonyl, ethyl sulfonyl and propyl sulfonyl.
The term "aryl" refers to an aromatic radical having from 6 to 14 carbon atoms such as phenyl, naphthyl, tetrahydronapthyl, indanyl, and biphenyl.
The term "arylalkyl" refers to an aryl group as defined above directly bonded to an alkyl group as defined above, e.g., -CH2C6H5 and -C2H5C6H5.
The terms "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. For purposes of this invention, the heterocyclic ring radical may be a monocyclic, bicyclic or tricyclic ring system, which may include fused, bridged or spiro ring systems, and the nitrogen, phosphorus, carbon, oxygen or sulfur atoms in the heterocyclic ring radical may be optionally oxidized to various oxidation states. In addition, the nitrogen atom may be optionally quaternized; Examples of such heterocyclic ring radicals include, but are not limited to, , tetrahydroisouinolyl, piperidinyl, piperazinyl, azapanyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxoazepinyl, azepinyl, pyrrolyl, 4-piperidonyl, pyrrolidinyl, pyrazinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, decahydroisoquinolyl, tetrahydrofurtyl, tetrahydropyranyl, thienyl, benzothienyl, thiamorpholinyl, thiamorpholinyl sulfoxide, thiamorpholinyl sulfone, dioxaphospholanyl, oxadiazolyl. The heterocyclic ring radical may be attached to the main structure at any heteroatom or carbon atom that results in the creation of a stable structure.
The term "bridged heterocyclyl" refers to unsaturated or saturated bridged cyclic system that is not aromatic. Such a system may contain isolated or conjugated unsaturation, but not aromatic or heteroaromatic rings in its core structure (but may have aromatic substitution thereon). And at least one ring in the system is inclusive of one or more heteroatoms, wherein each ring in the system contains 3 to 7 ring members, e.g., 1 to 10 carbon atoms and 1 to 3 heteroatoms selected from N, O, P or S, wherein the S or P is optionally substituted with one or more oxo to provide the group SO or S02, PO or PO2. Some non-limiting examples of bridged heterobicyclic ring system include (lR,5S)-8- azabicyclo[3.2.1]octane, (lR,5S)-3-azabicyclo[3.1.0]hexane or (lR,5S)-3,8- diazabicyclo[3.2.1]octane. The term "heterocyclylalkyl" refers to a heterocyclic ring radical directly bonded to an alkyl group. The heterocyclylalkyl radical may be attached to the main structure at any carbon atom in the alkyl group that results in the creation of a stable structure.
The term "heteroaryl" refers to an aromatic heterocyclic ring radical. The heteroaryl ring radical may be attached to the main structure at any heteroatom or carbon atom that results in the creation of a stable structure. Examples of such heteroaryl ring include, but are not limited to imidazolyl, pyrazolyl, pyrrolyl, lH-pyrrolo[2,3-b]pyridinyl, tetrazoyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolinyl, oxazolidinyl, triazolyl, isoxazolyl, isoxasolidinyl, thiazolyl, thiazolinyl, thiazolidinyl, isothiazolyl, quinuclidinyl, isothiazolidinyl, indolyl, isoindolyl, indolinyl, isoindolinyl, quinolyl, isoquinolyl, benzimidazolyl, thiadiazolyl, benzothiazolyl, benzooxazolyl, furyl,
"Substituted" refers to 1-3 substituents on the same position or on different positions with the same groups or different groups. Unless otherwise specified, the term "substituted" as used herein refers to substitution with any one or any combination of the following substituents: hydroxy, halogen, carboxyl, cyano, nitro, oxo (=0), thio (=S), substituted or unsubstituted alkyl, haloalkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted amino, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocyclylalkyl ring, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted heterocyclic ring.
The term "prodrug" means a compound that is transformed in vivo to yield a compound of formula (1) or a pharmaceutically acceptable salt, hydrate or solvate, or metabolite of the compound. The transformation may occur by various mechanisms, such as through hydrolysis in blood. A discussion of the use of prodrugs is provided by T. Higuchi and W. Stella, "Pro-drugs as Novel Delivery Systems," Vol. 14 of the A.C.S. Symposium Series, and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987.
The term "treating" or "treatment" of a state, disease, disorder or condition includes:
(1) preventing or delaying the appearance of clinical symptoms of the state, disease, disorder or condition developing in a subject that may be afflicted with or predisposed to the state, disease, disorder or condition but does not yet experience or display clinical or subclinical symptoms of the state, disease, disorder or condition; (2) inhibiting the state, disease, disorder or condition, i.e., arresting or reducing the development of the state, disease, disorder or condition or at least one clinical or subclinical symptom thereof; or
(3) relieving the state, disease, disorder or condition, i.e., causing regression of the state, disease, disorder or condition or at least one of its clinical or subclinical symptoms.
The benefit to a subject receiving treatment is either statistically significant or at least perceptible to the subject or to the physician.
The term "subject" includes mammals (especially humans) and other animals, such as domestic animals (e.g., household pets including cats and dogs) and non-domestic animals (such as wildlife).
The term "therapeutically effective amount" means the amount of a compound that, when administered to a subject for treating a state, disease, disorder or condition, is sufficient to effect such treatment. The "therapeutically effective amount" will vary depending on the compound, the state, disease, disorder or condition and its severity and the age, weight, physical condition and responsiveness of the subject receiving treatment.
The compounds of the present invention may form salts. Non-limiting examples of pharmaceutically acceptable salts forming part of this invention include salts derived from inorganic bases salts of organic bases salts of chiral bases, salts of natural amino acids and salts of non-natural amino acids. Certain compounds of the present invention are capable of existing in stereo isomeric forms (e.g., diastereomers, enantiomers, racemates, and combinations thereof). With respect to the overall compounds described by the formula (1), the present invention extends to these stereo isomeric forms and to mixtures thereof. To the extent prior art teaches synthesis or separation of particular stereoisomers, the different stereo isomeric forms of the present invention may be separated from one another by the methods known in the art, or a given isomer may be obtained by stereo specific or asymmetric synthesis. Tautomeric forms and mixtures of compounds described herein are also contemplated.
Pharmaceutically acceptable solvates includes hydrates and other solvents of crystallization (such as alcohols). The compounds of the present invention may form solvates with low molecular weight solvents by methods known in the art.
PHARMACEUTICAL COMPOSITIONS
The pharmaceutical compositions provided in the present invention include at least one compound described herein and at least one pharmaceutically acceptable excipient (such as a pharmaceutically acceptable carrier or diluent). Specifically, the contemplated pharmaceutical compositions include a compound(s) described herein in an amount sufficient to treat viral infection in a subject.
The 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.
Examples of suitable carriers include, but are not limited to, water, salt solutions, alcohols, polyethylene glycols, polyhydroxyethoxylated castor oil, peanut oil, olive oil, gelatin, lactose, terra alba, sucrose, dextrin, magnesium carbonate, sugar, cyclodextrin, amylose, magnesium stearate, talc, gelatin, agar, pectin, acacia, stearic acid or lower alkyl ethers of cellulose, silicic acid, fatty acids, fatty acid amines, fatty acid monoglycerides and diglycerides, pentaerythritol fatty acid esters, polyoxyethylene, hydroxymethylcellulose and polyvinylpyrrolidone.
The carrier or diluent may include a sustained release material, such as, for example, glyceryl monostearate or glyceryl distearate, alone or mixed with a wax.
The pharmaceutical composition may also include one or more pharmaceutically acceptable auxiliary agents, wetting agents, emulsifying agents, suspending agents, preserving agents, salts for influencing osmotic pressure, buffers, sweetening agents, flavoring agents, colorants, or any combination of the foregoing. The pharmaceutical composition of the invention may be formulated so as to provide quick-, sustained-, or delayed-release of the active ingredient after administration to the subject by employing procedures known in the art.
The pharmaceutical compositions described herein may be prepared, e.g., as described in Remington: The Science and Practice of Pharmacy, 20th Ed., 2003 (Lippincott Williams & Wilkins). For example, the active compound can be mixed with a carrier, or diluted by a carrier, or enclosed within a carrier, which may be in the form of an ampule, capsule, or sachet. When the carrier serves as a diluent, it may be a solid, semi- solid, or liquid material that acts as a vehicle, excipient, or medium for the active compound.
The pharmaceutical compositions may be, for example, capsules, tablets, aerosols, solutions, suspensions, liquids, gels, or products for topical application.
The route of administration may be any route which effectively transports the active compound to the appropriate or desired site of action. Suitable routes of administration include, but are not limited to, oral, nasal, pulmonary, buccal, subdermal, intradermal, transdermal, parenteral, rectal, depot, subcutaneous, intravenous, intraurethral, intramuscular, intranasal, ophthalmic (such as with an ophthalmic solution) or topical (such as with a topical ointment). The oral route is specifically suitable.
Solid oral formulations include, but are not limited to, tablets, capsules (soft or hard gelatin), dragees (containing the active ingredient in powder or pellet form), troches and lozenges. Tablets, dragees, or capsules having talc and/or a carbohydrate carrier or binder or the like are particularly suitable for oral application. Exemplary carriers for tablets, dragees, or capsules include lactose, cornstarch, and/or potato starch. A syrup or elixir can be used in cases where a sweetened vehicle can be employed.
A typical tablet that may be prepared by conventional tableting techniques.
Liquid formulations include, but are not limited to, syrups, emulsions, soft gelatin and sterile injectable liquids, such as aqueous or non-aqueous liquid suspensions or solutions.
For parenteral application, particularly suitable are injectable solutions or suspensions, specifically aqueous solutions with the active compound dissolved in polyhydroxylated castor oil.
METHODS OF SCREENING
Antiviral HIV activity and cytotoxicity of compounds present invention can be measured in parallel by following the methods published in the literature.
The cytotoxic effect of compounds can be analyzed by measuring the proliferation of cells using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazlium bromide (MTT) staining. Cells (5 x 10 cells /well) will be incubated in in 96 well plates in the presence or absence of compounds. At the end of treatment, 20μ1 of MTT (5mg/ml in PBS) will be added to each well and incubated for an additional 4 hours at 37°C. The purple -blue MTT formazan precipitate will be dissolved in a triplex reagent containing 10% SDS, 5% isobutanol and 10 mmol/lit HC1. The activity of mitochondria, reflecting cellular growth and viability, will be evaluated by measuring the optical density at 570 nm on micro titer plate.
Action of compounds on replication of HIV in Sup-Tl cells can be determined by the method published by Roda Rani et al., 2006 (Archives of Biochemistry and Biophysics, Volume 456, Issue 1, 1 December 2006, Pages 79-92).
Briefly, lxlO6 Sup-Tl cells with 100% cell viability will be seeded in RPMI 1640, 0.1% FBS four 12 well plates. Increasing concentrations of Epap-1 peptides will be added to the cells and will be infected with HIV1 93 I 101 each at final concentration of virus equivalent to 2 ng of p24 per ml. The infected cells will be incubated at 37 C and 5% C02 incubator for 2 hours. After 2hrs the cells will be pelleted at 350 g for 10 min, supernatant will be discarded and cell will be held with RPMI 1640 containing 10% FBS. The cells will be resuspended in the same medium with increasing concentrations of Epap-1 peptides and will be incubated for 96 hours. The cells will be supplemented with peptides at every 24 hours.The supernatants will be collected after 96 hours and analyzed using P24 antigen capture assay kit (SAIC Fredrick). The infection in the absence of Epap-1 will be considered to be 0% inhibition Azidothymidine (AZT) will be taken as positive control.
Action of compound on virus entry and quantification of virus entered can be done in terms of GFP expression by the following the methods published J. Virol. 72, 6988 (1998) by in Cecilia et al., and Analytical Biochemistry Volume 360, Issue 2, 15 January 2007, Pages 315-317 (Dyavar S. Ravi and Debashis Mitra).
Briefly, cells will be seeded in to wells of 24 well plates 1 day prior to the experiment. The cells will be transfected with Tat-reporter. The virus inoculum will be adjusted to 1,000- 4,000 TCID 50/ ml in assay medium (DMEM,10%FCS,glutamine and antibiotics), 50 μΐ aliquots will be incubated with serial dilutions of compounds (50 μΐ ) for lhr at 37°C. The reporter expression will be quantified at appropriate time calculated inhibitory doses referrers to the concentration of these agents in this preincubation mixture.
Other relevant references useful for screening antiviral HIV activity are: Averett,
D.R.1989. Anti-HIV compound assessment by two novel high capacity assays. J. Virol. Methods 23: 263-276; Schwartz, O., et al.1998; A rapid and simple colorimeric test fror the study of anti HIV agents. AIDS Res. and Human Retroviruses, 4(6):441-447; Daluge, S. M., et al. 1994. 5-Chloro-2',3'-deoxy-3'fluorouridine (935U83), a selective anti human immunodeficiency virus agent with an improved metabolic and toxicological profile; Antimicro. Agents and Chemotherapy, 38(7): 1590-1603; H.Mitsuya and S. Border, Inhibition of the in vitro infectivity and cytopathic effect of human T-lymphotropic virus type lymphadenopathy-associated virus (HLTV-III/LAV) by 2,3'-dideoxynucleosides, Proc. Natl. Acad. Sci. USA,83, 1911-15(1986); Pennington et al., Peptides 1990; Meek T.D et al., Inhibition of HIV-1 protease in infected T-limphocytes by synthetic peptide analogues, Nature, 343, p90 (1990); Weislow et al., J. Natl. Cancer Inst. 81, 577-586, 1989; T. Mimoto et al ., J. Med. Chem., 42, 1789-1802, 1999; Uckun et al 1998, Antimicobial Agents and Chemotherapy 42:383; for P24 antigen assay Erice et al., 1993, Antimicrob. Ag. Chemotherapy 37: 385-383; Koyanagi et al., Int. J. Cancer, 36, 445-451, 1985; Balzarini et al. AIDS (1991), 5, 21-28; Connor et al., Journal of virology, 1996, 70, 5306-5311; Popik et al., Journal of virology, 2002, 76, 4709-4722; Harrigton et al., Journal of Virology Methods, 2000, 88, 111-115; Roos et al.,Virology 2000, 273, 307-315; Fedyuk N.V. et al; Problems of Virology 1992, (3)P135; Mosmann T, December 1983, Journal of immunological methods, 65 (1-2), 55-63 ; SPC Cole, cancer chemotherapy and Pharmacology, 1986, 17, 259-263.
METHODS OF TREATMENT The present invention provides compounds and pharmaceutical 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. For example, PCT publication Nos. WO 01//07646, WO 01/65957, or WO 03/037908; US publication Nos. US 4,598,095 or US 2002/0068757; EP publication Nos. EP 0989862 or EP 0724650; Bioorganic & Medicinal Chemistry Letters, 16, (6), 1712-1715, 2006; and references cited therein, all of which are incorporated herein by reference in their entirety and for the purpose stated.
The present invention further provides a method of treating a disease, condition and/or disorder mediated by viral infections in a subject in need thereof by administering to the subject a therapeutically effective amount of a compound or a pharmaceutical composition of the present invention.
Diseases, conditions, and/or disorders that are mediated by viral infections are believed to include, but are not limited to, FHV infection, HBV, HCV, a retroviral infection genetically related to HIV, AIDS, inflammatory disease, respiratory disorders (including adult respiratory distress syndrome (ARDS), bronchitis, chronic bronchitis, chronic obstructive pulmonary disease, cystic fibrosis, asthma, emphysema, rhinitis and chronic sinusitis), inflammatory bowel disease (including Crohn's disease and ulcerative colitis), multiple sclerosis, rheumatoid arthritis, graft rejection (in particular but not limited to kidney and lung allografts), endometriosis, type I diabetes, renal diseases, chronic pancreatitis, inflammatory lung conditions, chronic heart failure and bacterial infections (in particular but not limited to tuberculosis).
The compounds of the present invention can obtain more advantageous effects than additive effects in the prevention or treatment of the above diseases when using suitably in combination with the available drugs. Also, the administration dose can be decreased in comparison with administration of either drug alone, or adverse effects of co administrated drugs other than antiviral can be avoided or declined.
METHODS OF PREPARATION
The compounds described herein may be prepared by techniques known in the art. In addition, the compounds described herein may be prepared by following the reaction sequence as depicted in Scheme- 1 to 3. Further, in the following schemes, where specific bases, acids, reagents, solvents, coupling agents, etc., are mentioned, it is understood that other bases, acids, reagents, solvents, coupling agents etc., known in the art may also be used and are therefore included within the present invention. Variations in reaction conditions, for example, temperature and/or duration of the reaction, which may be used as known in the art, are also within the scope of the present invention. All the stereoisomers of the compounds in these schemes, unless otherwise specified, are also encompassed within the scope of this invention.
In a further aspect, the compounds of the present invention can also contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the present invention also embraces isotopically-labeled variants of the present invention which are identical to those recited herein, but for the fact that one or more atoms of the compound are replaced by an atom having the atomic mass or mass number different from the predominant atomic mass or mass number usually found in nature for the atom. All isotopes of any particular atom or element as specified are contemplated within the scope of the compounds of the invention, and their uses. Exemplary isotopes that can be incorporated in to compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, sulfur, fluorine, chlorine and iodine, such as H ("D"), 3H, nC, 13C, 14C, 13N, 15N, 150, 170, 180, 32P, 33P, 35S, 18F, 36C1, 123I and 125I. Particular isotopes are -CD3 or -C(D2)-. Isotopically labeled compounds of the present inventions can generally be prepared by following procedures analogous to those disclosed in the Schemes and/or in the Examples herein below, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.
Compounds of the present invention can be synthesized from naturally occurring Betulinic acid or betulinal. Key intermediates required for synthesizing analogues are either commercially available, or can be prepared by the methods published in the literature. For example, the key intermediates in the present invention were prepared by modifying the procedures published in Journal of organic chemistry 2013, 78, 1463-1470; Organic Letters, 2009, 11, 3270-3273; Organic Letters, 2007, 9, 3675-3678; Bio Organic Medicinal Chemistry Letters, 2011, 21, 562-568.
Scheme-1
Figure imgf000030_0001
The compounds of formula 5 (R7, R8, Zi, Z2 and Z3 are same as defined above) can be prepared as described in Scheme 1. The nitro compound of formula 1 can be reacted with the aldehydes of formula 2 followed by substituted N-contained heterocycles of formula 3 to give the compounds of formula 4 in the presence of base solutions such as sodium hydroxide (NaOH), potassium hydroxide (KOH) or the like. The compounds of formula 4 can be reduced with reducing agents such as raney nickel, palladium in carbon (Pd/C) or the like to give the compounds of formula 5 in the presence of suitable solvents such as methanol (MeoH), ethylacetate (EtOAc) or the like.
Scheme-2
Figure imgf000031_0001
N-
/ :
The compounds of formula 12 & 14 ( '·-—-'' , Zi, Z2 and Z3 are same as defined above) can be prepared as described in scheme 2. The prolinal compounds of formula 6 can be converted to N-protected compounds of formula 7 in the presence of protecting groups such as tert-butyloxy carbonyl (BOC), benzyl, carboxybenzyl (cbz) or the like in the presence of solvents such as dichloromethane (DCM), Ν,Ν-dimethyl formamide (DMF), tetrahydrofuran (THF) or the like. The compounds of formula 7 can be converted to compounds of formula 8 in the presence of bases such as lithium aluminum hydride (LAH), sodiumborohydride (NaBH4), boron in tetrahydrofuran or the like in the solvents such as tetrahydrofuran (THF), ether, ethylacetate (EtOAc) or the like. The compounds of formula 8 can be converted to O- protected compounds of formula 9 in the presence of protecting groups such as p-toluene sulfonyl chloride, benzyl, i-Butyldimethylsilyl (TBDMS), benzoyl or the like in the solvents such as dichloromethane (DCM), tetrahydrofuran (THF) or the like. The compounds of formula 9 coupled with compounds of formula 10 can be converted to compounds of formula 11 in the presence of solvents such as 1,4-dioxane, tetrahydrofuran (THF), toluene or the like in presence of without base and with bases such as sodium hydroxide (NaOH), sodium bicarbonate (NaHC03), sodium carbonate (Na2C03) or the like. The compounds of formula
11 can be converted to compounds of formula 12 in the presence of deprotecting reagents such as HC1, trifluoro acetic acid (TFA) or the like in the solvents such as dichloromethane (DCM), dioxane or the like. The compounds of formula 7 can be coupled with the compounds of formula 10 to give the compounds of formula 13 in the presence of l-Ethyl-3- (3-dimethylaminopropyl)carbodiimide (EDCI), N-Hydroxybenzotriazole (HOBt) or the like in the solvents such as Ν,Ν-dimethyl formamide (DMF), N,N-Diisopropylethylamine (DIPEA) or the like. The compounds of formula 13 can be deprotected in the presence deprotecting reagents such as HC1, trifluoro acetic acid (TFA) or the like in the solvents such as dichloromethane (DCM), dioxane or the like to give the compounds of formula 14.
Scheme - 3
Figure imgf000032_0001
[Formula (I), when Y=CH2 and n=0]
The compounds of formula (I) (wherein, Ri, R2, R3, R4, R5, R6, R7, X, Zi, Z2 and Z3 are same as defined above) can be prepared as described in Scheme 3. The compounds of formula 15 (as described in our PCT publication WO 2013/160810) can be converted to the halide compounds of formula 16 in the presence of halogenating agents such as thionyl chloride (SOCl2), oxalyl chloride (COCl2), phosphorous bromide, phosphorous oxy bromide, phosphorous pentachloride, phosphorous tribromide, phosphorous pentabromide or the like in the solvents such as benzene, toluene, dichloromethane (DCM) or the like. The compounds of formula 16 can be converted to amide compounds of formula 17 with their corresponding amine compounds of formula 5 in the presence of bases such as triethylamine (TEA), N,N- diisopropylethylamine (DIPEA), pyridine or the like in the solvents such as N,N- di.methylfonnam.ide (DMF), dichloromethane (DCM), tetrahydrofuran (THF) or the like. Alternately, C28-amide compounds of formula 17 can also be prepared by using suitable coupling agents such as l-ethyl-3-(3-dimethylaminopropyl) carbodiimide) hydrochloride (EDCI), (l-[Bis(dimethylamino)methylene]-lH-l,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate) (HATU), 1- Hydroxybenzotriazole (HOBt) monohydrate or the like with suitable bases such as tri ethyl amine (TEA), Ν,Ν-diisopropylethy famine (DIPEA), pyridine or the like with nicotinykhloride or benzoylchioride in the solvents such as N,N- dimethylfomi amide (DM F), dichloromethane (DCM), tetrahydrofuran (TBF) or the like. The compounds of formula 17 can be deprotected in the presence of bases such as potassium carbonate, sodium hydroxide, ammonia or the like in the solvents such as methanol: tetrahydrofuran, methanol: water, methanol or the like to give the C3 hydroxy compounds of formula 18. The C3 hydroxy compounds of formula 18 can be reacted with corresponding acid anhydrides, half protected diacids or their mixed anhydrides or acid chlorides to give the corresponding acid compounds of the present invention formula (1) in the presence a bases such as tri ethyl amine (TEA), 4-dimethylaminopyridine, diisopropyl. ethyl mine or pyridine or the like in the solvents such as dichloromethane (DCM), toluene, ethylacetate (EtOAc), tetrahydrofuran (THF) or the like.
The abbreviations used in the entire specification may be summarized herein below with their particular meaning: DIPEA (Ν,Ν-Diisopropylethylamine); °C (degree Celsius); δ (delta); ppm (parts per million); % (percentage); DMSO-d6 (Deuterated DMSO); d (Doublet); dd (Doublet of doublet); EtOH (Ethanol); EtOAc (Ethyl acetate); g or gr (gram); H or H2 (Hydrogen); HC1 (Hydrochloric acid); h or hr. (Hours); HATU (0-(7-Azabenzotriazol-l-yl)- Ν,Ν,Ν',Ν'-tetramethyluroniumhexafluoro phosphate); Hz (Hertz); HPLC (High-performance liquid chromatography); mmol (Milli mol); M (Molar); ml (Millilitre); mg (Milli gram); m (Multiplet); mm (Millimetre); MHz (Megahertz); ESI-MS (Electron spray Ionization Mass spectra); min (Minutes); mM (Milli molar); NaOH (Sodium hydroxide); N2 (Nitrogen); NMR (Nuclear magnetic resonance spectroscopy); S (Singlet); TEA (Triethyl amine); TLC (Thin Layer Chromatography); THF (Tetrahydrofuran); tert (Tertiary), t (Triplet); IC (Inhibitory concentration), nM (Nano molar); pH (Pouvoir hydrogen); (Boc)20 (Di-tert-butyl dicarbonate); DCM (dichloromethane); DMF (Ν,Ν-dimethyl formamide); DMAP (4- (Dimethylamino)pyridine); eq (equivalent); Ltr or L (Liter); CDC13 (Deuterated chloroform); J (Coupling constant); AcOH (Acetic acid); ABq (AB quartet); K2C(¾ (potassium carbonate); Cs2C03 (Cesium carbonate); NaHC03 (Sodium bicarbonate); Na2S04 (Sodium sulphate); MeOH (methanol); EDCI (l-Ethyl-3-(3-dimethylaminopropyl)carbodiimide); HOBt (1 -Hydroxybenzotriazole); brs (broad singlet); DCC (Ν,Ν'-dicyclohexylcarbodiimide); Pd/C (palladium in carbon) and Pd(OAc)2 (palladium acetate).
EXPERIMENTAL The present invention is further illustrated by the following examples, which are not to be construed in any way as imposing limitations upon the scope of this disclosure, but rather are intended to be illustrative only. On the contrary, it is to be clearly understood that resort may be had to various other embodiments, modifications, and equivalents thereof which, after reading the description herein, may suggest themselves to one of ordinary skill in the art without departing from the spirit of the present invention. Thus, the skilled artisan will appreciate how the experiments and examples may be further implemented as disclosed by variously altering the following examples, substituents, reagents, or conditions.
INTERMEDIATES
Intermediate 1: Preparation of 2-methy -l-morpholinopropan-2-amine:
Figure imgf000034_0001
Step 1: Synthesis of4-(2-methyl-2-nitropropyl)morpholine:
Figure imgf000034_0002
To the stirred solution of morpholine (3 g, 34.48 mmol, 1.0 eq) and 2-nitropropane (2.0 g, 34.48 mmol, 1.0 eq) are cooled with ice bath then added formaldehyde (37%) (1.03 mL, 34.48 mmol, 1.0 eq) and NaOH solution (0.5 mol/L) are slowly added dropwise at 10°C then the reaction mixture was stirred for about 1 hour at 25 °C and about 1 hour at 50 °C. After completion of the reaction (monitored by TLC), the solution is treated with water, ether and aqueous phase is extracted with water. The combined organic phase is dried over NaS04, and combined with HC1 in dioxane (mol/1). The precipitate formed which was filtered and dried under vacuum to afford the desired product (4.0 g, yield: 62.0%) as a white powder. 1H NMR (CD3OD, 300 MHz): δ 3.60 (m, 4H), 2.52 (m, 4H), 2.37 (m, 2H), and 1.18 (s, 6H); Mass: [M]+ 189.18 (100%).
Step 2: Synthesis of 2 -methyl- l-morpholinopropan-2 -amine:
To the stirred solution of 4-(2-methyl-2-nitropropyl)morpholine (step 1, 4.0 g, 21.27 mmol, 1.0 eq) in methanol (20 mL) then added Raney Nickel (1.7 g, 29.79 mmol, 1.4 eq) treated with hydrogen (50 psi) at room temperature for about 1 hour. After completion of the reaction (Monitored by TLC), the reaction mixture was filtered through celite bed and the filtrate was concentrated to afford the desired product (3.2 g, yield: 95.0%) as a light yellow solid. 1H NMR (CD3OD, 300 MHz): δ 7.75 (s, 2H), 3.60 (m, 4H), 2.52 (m, 4H), 2.37 (m, 2H), and 1.18 (s, 6H); Mass: [M]+ 159.18 (100%). The below intermediates 2-6 were prepared by the procedure similar (including reagents and reaction conditions) to the above described in the synthesis of intermediate- 1 by using suitable reagents, reactents and solvents at appropriate conditions.
Figure imgf000035_0002
Intermediate 7: Preparation of l-(4-benzylpiperazin-l-yl)-2-methylpropan-2-amine:
H2NJ^ ~^
Step 1: Synthesis of tert-butyl 4-benzylpiperazine-l-carboxylate:
Figure imgf000035_0001
To the stirred solution of tert-butyl piperazine-l-carboxylate (6.0 g, 33.0 mmol, 1.0 eq) in acetonitrile (60 niL), the reaction mixture was cooled at 0 °C, then added triethylamine (22 mL, 161 mmol, 5.0eq) and (bromomethyl)benzene (6.0 mL, 35.0 mmol, 1.1 eq) drop wise. The reaction mixture was stirred at room temperature for about 6 hours. The reaction mixture was diluted with water and extracted with CH2CI2. The combined organic extracts were dried over Na2S04, filtered and evaporated under reduced pressure to afford the desired product (6.5 g, yield: 76.0%) as a white solid.
Step 2: Synthesis of 1-benzylpiperazine:
Figure imgf000036_0001
To the stirred solution of tert-butyl 4-benzylpiperazine-l-carboxylate (step 1, 6.5 g, 18.5 mmol, 1.0 eq) in 1,4-dioxane in HCl and stirred at room temperature for about 3 hours. The reaction mixture was evaporated under reduced pressure to afford the desired product (4.0 g) as a white solid.
Step 3: Synthesis of 1 -benzyl-4-(2-methyl-2-nitropropyl)piperazine :
Figure imgf000036_0002
To the stirred solution of 1-benzylpiperazine (step 2, 4.0 g, 29.1 mmol, 1.0 eq), 2- nitropropane (2.8 mL, 29.1 mmol, 1.0 eq) are cooled with ice bath, then added formaldehyde (37%) (2.5 mL, 70.5 mmol, 3.0 eq) and NaOH solution (5.0 mL, 0.5 mol/L) are slowly dropwise at 10 °C. Then the reaction mixture was stirred for about 1 hour at 25 °C and about 1 hour at 50 °C. The solution is treated with water, ether and aqueous phase is extracted with water. The combined organic phase is dried over NaS04 and combined with HCl in dioxane (30 mL). Then the precipitate formed which was filtered and dried in vacuum to afford the desired product (2.0 g, yield: 35.0%) as a white solid. 1H NMR (CD3OD, 300 MHz): δ 7.30- 7.22 (m, 5H), 3.41 (s, 2H), 2.79 (s, 2H), 2.46 (t, 4H), 2.29 (t, 4H) and 1.47 (s, 6H); Mass: [M]+ 278.25 (100%).
Step 4: Synthesis of 1 -(4-benzylpiperazin-l -yl)-2-methylpropan-2-amine:
To the stirred solution of l-benzyl-4-(2-methyl-2-nitropropyl)piperazine (step 3, 2.0 g, 7.3 mmol, 1.0 eq) in Methanol (30 mL), then added Raney Nickel (0.200 g, 4.8 mmol, 0.6 eq) and treated with hydrogen (50 psi) at room temperature for about 12 hours. The reaction mixture was filtered through celite and filtrate was concentrated to obtain the desired product (1.2 g, yield: 65%) as a white powder. 1H NMR (CD3OD, 300 MHz): δ 7.30-7.22 (m, 5H), 3.43 (s, 2H), 3.07 (s, 2H), 2.35 (bs, 4H), 2.13 (t, 4H) and 0.93 (s, 6H); Mass: [M]+ 248.16
(100%).
Intermediate 8: Preparation of (R)-l-(3-(2-methoxyethoxy)pyrrolidin-l-yl)-2-methylpropan- 2-amine:
Figure imgf000037_0001
Step 1: Synthesis of tert-butyl (R)-3-hydroxypyrrolidine-l-carboxylate:
H
Figure imgf000037_0002
To a stirred solution of (R)-pyrrolidin-3-ol hydro chloride (3.0 g, 24.2 mmol) in DCM (30 ml), were added N(Et)3 (10.2 ml, 72.8 mmol) and (Boc)20 (6 ml, 26.7 mmol) at 0 °C. The reaction mixture was allowed to stir at room temperature for about 12 hours. After 12 hours of stirring, saturated NH4C1 was added. The solution was extracted with EtOAc (2x60 mL). The combined organic phases were washed with brine, dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by flash chromatography with EtOAc-hexane (4: 6) as an eluent to afford the desired product (4.5 g, yield: 99%) as an oil.
Step 2: Synthesis of tert-butyl (R)-3- 2-methoxyethoxy)pyrrolidine-l-carboxylate:
Figure imgf000037_0003
To a stirred solution of NaH (3.8 g, 95 mmol, 60% in mineral oil w/w) in THF (20 ml) under N2 atmosphere at 0 °C, was added tert-butyl (R)-3-hydroxypyrrolidine-l- carboxylate (step 1, 4.5 g, 24 mmol) in THF (40 ml). After 30 minutes l-bromo-2- methoxyethane (3.4 ml, 36 mmol) was added, the reaction mixture was slowly allowed to attain to room temperature and stirred for overnight. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by addition of saturated NH4C1 solution at 0 °C. The solution was extracted with EtOAc (2x100 mL), the combined organic phases were washed with brine, dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by flash chromatography with EtOAc-hexane (2: 8) as an eluent to afford the desired product (3.0 g, yield: 51%) as an oil. 1H MR (300 MHz, CDC13): δ 4.10-4.05 (m, 1H), 3.60-3.52 (m, 4H), 3.45-3.36 (m, 4H), 3.38 (s, 3H), 2.02-1.93 (m, 2H), 1.45 (s, 9H).
Step 3: Synthesis of (R)-3-(2-methoxyethoxy)-l-(2-methyl-2-nitropropyl)pyrrolidine:
>c OMe
N0 36"°' To a stirred solution of tert-butyl 4-(2-methoxyethoxy)piperidine-l-carboxylate (step
2, 3.0 g, 12.3 mmol) in dioxane (10 ml), was added 6N HC1 in dioxane (30 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product 4-(2-methoxyethoxy) piperidine hydrochloride (2.1 g, yield: 100 %). Next reaction was carried out without any further purification.
To a stirred solution of 4-(2-methoxyethoxy)piperidine hydrochloride (2.0 g, 11 mmol) in dioxane (10 ml), were added 2-nitropropane (1.5 ml, 16.5 mmol), formaldehyde solution (1.5 ml 30% in H20, 16.4 mmol), IN NaOH solution (3 ml) at 0 °C and stirring was continued for about 1 hour at 0 °C. The reaction mixture temperature was raised to 60 °C and continued for about 5 hours. After completion of the reaction (monitored by TLC), the solution was extracted with EtOAc (2x60 mL), the combined organic phases were washed with brine, dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by flash chromatography with EtOAc-hexane (7: 3) as an eluent to afford the desired product (2.0 g, yield: 77%) as an oil. 1H NMR (300 MHz, CDC13): δ 4.02- 3.97 (m, 1H), 3.52-3.51 (m, 4H), 3.37 (s, 3H), 2.99 (d, J = 2.4 Hz, 2H), 2.96-2.91 (m, 1H), 2.74-2.60 (m, 3H), 2.03-1.91 (m, 1H), 1.83-1.75 (m, 1H), 1.56 (s, 6H).
Step 4: Synthesis of (R)-l-(3-(2-methoxyethoxy)pyrrolidin-l-yl)-2-methylpropan-2-amine:
To a stirred solution of (R)-3-(2-methoxyethoxy)-l-(2-methyl-2-nitropropyl) pyrrolidine (step 3, 2.0 g, 8.13 mmol) in methanol (20 ml) was added 10% Pd/C (0.20 g) and purged with nitrogen. The reaction mixture was stirred for about 12 hours under H2 atmosphere. After completion of the reaction (monitored by TLC), the reaction mixture was filtered and filtrate was concentrated under reduced pressure to give the desired product. Next reaction was carried out without any further purification.
Intermediate 9: Preparation of l-(4-(2-methoxyethoxy)piperidin-l-yl)-2-methylpropan-2- amine:
Η,Ν^ ^\ . \ ^OMe
Step 1: Synthesis of tert-butyl 4-oxopiperidine-l -carboxylate:
Figure imgf000038_0001
To a stirred solution of piperidine-4-one hydrochloride (10 g, 65 mmol) in DCM (120 ml), were added N(Et)3 (27 ml, 195 mmol) and (Boc)20 (15 ml, 65 mmol) at 0 °C. The reaction mixture was allowed to stir at room temperature for about 8 hours. After 8 hours of stirring, saturated NH4C1 was added. The solution was extracted with EtOAc (2x150 niL), the combined organic phases were washed with brine, dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by flash chromatography with EtOAc-hexane (3:7) as an eluent to afford the desired product (12 g, yield: 92%) as a colourless oil. 1H NMR (300 MHz, DMSO-d6): δ 3.60 (t, J= 6.3 Hz, 4H), 2.34 (t, J= 6.3 Hz, 4H), 1.42 (s, 9H).
Step 2: Synthesis of tert-butyl 4-hydroxypiperidine-l-carboxylate:
Figure imgf000039_0001
To a stirred solution of tert-butyl 4-oxopiperidine-l-carboxylate (step 1, 12.95 g, 65 mmol) in MeOH (130 ml), was slowly added NaBH4 (12.3 g, 325 mmol) over about 15 minutes (Care - vigorous effervescence) at 0 °C. The reaction mixture was allowed to stir at room temperature for about 5 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by addition of saturated NH4C1 solution at 0°C. The solution was extracted with EtOAc (2x150 mL), the combined organic phases were washed with brine, dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by flash chromatography with EtOAc-hexane (4:6) as an eluent to afford the desired product (11 g, yield: 84%) as a colourless oil. 1H NMR (300 MHz, DMSO- d6): δ 4.69 (d, J= 4.2 Hz, 1H), 3.68-3.58 (m, 3H), 2.97-2.90 (m, 2H), 1.69-1.63 (m, 2H), 1.38 (s, 9H), 1.28-1.16 (m, 2H).
Step 3: Synthesis of tert-butyl 4-(2-methoxyethoxy)piperidine-l-carboxylate:
Figure imgf000039_0002
To a stirred solution of NaH (3.3 g, 52 mmol, 60% in mineral oil w/w) in THF (20 ml) under N2 atmosphere at 0 °C, was added tert-butyl 4-hydroxypiperidine-l-carboxylate (step 2, 5.2 g, 26 mmol) in THF (30 ml). After 30 minutes 1 -bromo-2-methoxy ethane (3.7 ml, 39 mmol) was added. The reaction mixture was slowly allowed to attain to room temperature and stirred for overnight. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by addition of saturated NH4C1 solution at 0 °C. The solution was extracted with EtOAc (2x100 mL), combined organic phases were washed with brine, dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by flash chromatography with EtOAc-hexane (2: 8) as an eluent to afford the desired product (4.0 g, yield: 59%) as a colorless oil.1H NMR (300 MHz, DMSO-d6): δ 3.65-3.57 (m, 2H), 3.54-3.51 (m, 2H), 3.47-3.40 (m, 3H), 3.24 (s, 3H), 3.03-2.94 (m, 2H), 1.79-1.73 (m, 2H), 1.38 (s, 9H), 1.36-1.23 (m, 2H).
Step 4: Synthesis of 4-(2-methoxyethoxy)-l -(2-methyl-2-nitropropyl)piperidine:
Figure imgf000040_0001
To a stirred solution of tert-butyl 4-(2-methoxyethoxy)piperidine-l-carboxylate (step 3, 3.93 g, 15.3 mmol) in dioxane (10 ml), was added 6N HC1 in dioxane (30 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product 4-(2- methoxyethoxy)piperidine hydrochloride (2.95 g, yield: 100%). Next reaction was carried out without any further purification.
To a stirred solution of 4-(2-methoxyethoxy)piperidine hydrochloride (2.95 g, 15.1 mmol) in dioxane (10 ml), were added 2-nitropropane (2 ml, 22 mmol), formaldehyde solution (2.2 ml 30% in H20, 22 mmol), IN NaOH solution (4 ml) at 0 °C and stirring was continued for about 1 hour at 0 °C. The reaction mixture temperature was raised to 60 °C and continued for about 5 hours. After completion of the reaction (monitored by TLC), the solution was extracted with EtOAc (2x60 mL), combined organic phases were washed with brine, dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by flash chromatography with EtOAc-hexane (6:4) as an eluent to afford the desired product (1.6 g, yield: 44%) as an oil. 1H NMR (300 MHz, DMSO-d6): δ 3.49-3.45 (m, 2H), 3.41-3.38 (m, 2H), 3.25-3.18 (m, 1H), 3.23 (s, 3H), 2.78 (s, 2H), 2.63-2.57 (m, 2H), 2.30-2.22 (m, 2H), 1.74-1.73 (m, 2H), 1.47 (s, 6H), 1.38-1.31 (m, 2H).
Step 5: Synthesis of 1 -(4-(2-methoxyethoxy)piperidin-l -yl)-2-methylpropan-2 -amine:
To a stirred solution of 4-(2-methoxyethoxy)-l-(2-methyl-2-nitropropyl)piperidine (step 4, 1.6 g, 6.6 mmol) in methanol (20 ml) was added 10% Pd/C (0.20 g) and purged with nitrogen. The reaction mixture was stirred for about 12 hours under H2 atmosphere. After completion of the reaction (monitored by TLC), the reaction mixture was filtered and filtrate was concentrated under reduced pressure to give the desired product (1.3 g, yield: 85%) as an oil. 1H NMR (300 MHz, DMSO-d6): δ 3.49-3.48 (m, 2H), 3.42-3.39 (m, 2H), 3.25-3.18 (m, 1H), 3.23 (s, 3H), 2.74-2.70 (m, 2H), 2.31-2.23 (m, 2H), 2.12 (s, 2H), 1.78-1.75 (m, 2H), 1.48-1.36 (m, 2H), 0.97 (s, 6H).
Intermediate 10: Preparation of l-(4-ethoxy-4-phenylpiperidin-l-yl)-2-methylpropan-2- amine:
Figure imgf000040_0002
Step 1: Synthesis of tert-butyl 4-hydrox -4-phenylpiperidine-l -carboxylate :
Figure imgf000041_0001
To a stirred solution of Mg (0.12 g, 10.1 mmol) in THF (4 ml), were added I2 (cat), bromo benzene (2.3 ml, 15.1 mmol) and stirred for about 30 minutes. Then the reaction mixture was added a solution of tert-butyl 4-oxopiperidine-l -carboxylate (Intermediate 9-step 1, 2 g, 10.1 mmol) in THF (10 ml) and stirred for overnight. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by cautious addition of saturated NH4C1 solution at 0 °C and extracted with EtOAc (2x50 ml). The combined organic layers were washed with brine, dried over anhydrous Na2S04 and concentrated under reduced pressure to get the residue. The crude product was purified via silica gel column chromatography with EtOAc and n-Hexane (30:70) as an eluent to afford the desired compound (1.2 g, yield: 44.4%) as a thick oil. 1H MR (300 MHz, DMSO-d6): δ 7.48 (m, 2H), 7.34-7.29 (m, 2H), 7.23-7.18 (m, 1H), 3.86-3.82 (m, 2H), 3.11-3.08 (m, 2H), 1.83-1.73 (m, 2H), 1.59-1.55 (m, 2H), 1.41 (s, 9H).
Step 2: Synthesis of 4-ethoxy-l -(2-methyl-2-nitropropyl)-4-phenylpiperidine:
Figure imgf000041_0002
To a stirred solution of NaH (0.52 g, 12 mmol, 60% in mineral oil w/w) in DMF (3 ml) under N2 atmosphere at 0 °C, was added tert-butyl 4-hydroxy-4-phenylpiperidine-l- carboxylate (step 1, 1.2 g, 4.3 mmol) in DMF (15 ml). After 30 minutes ethyl iodide (0.5 ml, 6.4 mmol) was added, the reaction mixture was slowly allowed to attain to room temperature and stirred for overnight. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by addition of saturated NH4C1 solution at 0°C. The solution was extracted with EtOAc (2x100 niL), the combined organic phases were washed with brine, dried over Na2S04 and concentrated under reduced pressure to give tert-butyl 4-ethoxy-4- phenylpiperidine- 1 -carboxylate (1.1 g, yield: 92%) as an oil. The next reaction was carried out without any further purification.
To a stirred solution of tert-butyl 4-ethoxy-4-phenylpiperidine-l -carboxylate (1.1 g, 3.6 mmol) in dioxane (5 ml), was added 6N HC1 in dioxane (15 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford 4-ethoxy-4-phenylpiperidine hydrochloride (0.869 g, yield: 100%). Next reaction was carried out without any further purification. To a stirred solution of 4-ethoxy-4-phenylpiperidine hydrochloride (0.6 g, 4.1 mmol) in dioxane (3 ml), were added 2-nitropropane (0.6 ml, 6.1 mmol), formaldehyde solution (0.6 ml 30% in H20, 6.1 mmol), IN NaOH solution (0.5 ml) at 0 °C and stirring was continued for about 1 hour at 0 °C. The reaction temperature was raised to 60 °C and continued for about 5 hours. After completion of the reaction (monitored by TLC), the solution was extracted with EtOAc (2x15 mL), the combined organic phases were washed with brine, dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by flash chromatography with EtOAc-hexane (3:7) as an eluent to afford the desired product (0.3 g, yield: 39.4%) as an oil. 1H MR (300 MHz, DMSO-d6): δ 7.36-7.25 (m, 5H), 3.01-2.93 (q, J = 6.9 Hz, 2H), 2.85 (s, 2H), 2.68-2.35 (m, 4H), 1.87-1.78 (m, 4H), 1.50 (s, 6H), 1.03 (t, J= 6.9 Hz, 3H).
Step 3: Synthesis of 1 -(4-ethoxy-4-phenylpiperidin-l -yl)-2-methylpropan-2-amine:
To a stirred solution of 4-ethoxy-l-(2-methyl-2-nitropropyl)-4-phenylpiperidine (step 2, 0.3 g, 0.9 mmol) in methanol (8 ml) was added 10% Pd/C (0.03 g) and purged with nitrogen. The reaction mixture was stirred for about 12 hours under H2 atmosphere. After completion of the reaction (monitored by TLC), the reaction mixture was filtered and filtrate was concentrated under reduced pressure to give the desired product (0.15 g, yield: 72.5%) as a gummy oil. Next reaction was carried out without any further purification.
Intermediate 11: Preparation of (R)-l-(3-(benzyloxy)pyrrolidin-l-yl)-2-methylpropan-2- amine:
Bn
Figure imgf000042_0001
Step 1: Synthesis of tert-butyl (R)-3 -(benz loxy pyrrolidine- 1-carboxylate:
Figure imgf000042_0002
To the stirred solution of tert-butyl (R)-3-hydroxypyrrolidine-l-carboxylate (Intermediate 8-step 1, 3 g, 16.04 mmol, 1.0 eq) in DMF (30 mL), then the reaction mixture was cooled at 0 °C and added NaH (1.9 g, 80.2 mmol, 5.0 eq) followed by benzyl bromide (3.13 mL, 19.24 mmol, 1.2 eq). The reaction mixture was stirred at room temperature for about 3 hours. The reaction mixture was diluted with water and extracted with CH2C12. The combined organic extracts were dried over Na2S04, filtered and evaporated under reduced pressure. The residue was purified by column chromatography by using 10% ethylacetate and hexane as an eluent to afford the desired product (3.0 g, yield: 68.0%) as a white solid.
Step 2: Synthesis of ' (R)-3-(benzyloxy)pyrrolidine: , ΟΒΠ
HN
To the stirred solution of tert-butyl (R)-3-(benzyloxy)pyrrolidine-l-carboxylate (step 1) in 1,4-dioxane in HC1 and stirred at room temperature for about 3 hours. The reaction mixture evaporated under reduced pressure to afford the desired product (2.0 g) as a white solid.
Step 3: Synthesis of (R)-3 -(benzyloxy)-l -(2-methyl-2-nitropropyl)pyrrolidine :
Bn
Figure imgf000043_0001
To the stirred solution of (R)-3-(benzyloxy)pyrrolidine (step 2, 2.0 g, 11.29 mmol, 1.0 eq) and 2-nitropropane (1.2 mL, 13.55 mmol, 1.2 eq) are cooled with ice bath then added formaldehyde (37%) (2.1 mL, 27.09 mmol, 2.4 eq) and NaOH solution (2.0 mL, 0.5 mol/L) are slowly dropwise at 10 °C. Then the reaction mixture was stirred for about 1 hour at 25 °C and about 1 hour at 50 °C. The solution is treated with water, ether and aqueous phase is extracted with water. The combined organic phase is dried over NaS04, combined with HC1 in dioxane (30 mL), the precipitate formed which was filtered and dried in vacuum to afford the desired product (2.0 g, yield: 64.0%) as a white solid. 1H NMR (CD3OD, 300 MHz): δ 7.28-7.21 (m, 5H), 4.43 (q, 2H), 4.02-3.98 (m, 1H), 2.92 (m, 2H), 2.77 (m, 4H), 1.96-1.86 (m, 1H), 1.83-1.74 (m, 1H) and 1.53 (s, 6H); Mass: [M]+ 279.14 (100%).
Step 4: Synthesis of (R)-l -(3-(benzyloxy)pyrrolidin-l -yl)-2-methylpropan-2-amine:
To the solution of (R)-3-(benzyloxy)-l-(2-methyl-2-nitropropyl)pyrrolidine (step 3, 2.0 g, 7.19 mmol, 1.0 eq) in Methanol (30 mL) then added Raney Nickel (1.0 g, 35.89 mmol, 1.4 eq) and treated with hydrogen (50 psi) and stirred at room temperature for about 1 hour. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through celite and filtrate was concentrated to afford the desired product (1.2 g, yield: 70%) as a white powder. 1H NMR (CD3OD, 300 MHz): δ 7.30 (s, 2H), 7.28-7.21 (m, 5H), 4.43 (q, 2H), 4.02-3.98 (m, 1H), 2.92 (m, 2H), 2.77 (m, 4H), 1.96-1.86 (m, 1H), 1.83-1.74 (m, 1H) and 1.53 (s, 6H); Mass: [M]+ 249.56 (100%).
Intermediate 12: Preparation of 2-methyl-l-(4-(methylsulfonyl)piperazin-l-yl)propan-2- amine:
Figure imgf000043_0002
Step 1: Synthesis of tert-butyl piperazine-1 -carboxylate:
Figure imgf000044_0001
Piperazine (20.0 g, 232.5 mmol, 1.0 eq) dissolved in DCM (400 mL) which was stirred at 0 °C then added (Boc)20 (25.3 mL, 116 mmol, 0.5 eq) in DCM (400 mL) was added over about 30 minutes and was stirred at room temperature for overnight. After completion of the reaction, the reaction mixture was cooled to 0 °C for about 15-20 minutes. The precipitated solid was filtered and the filtrate was concentrated to half of its volume after which, the DCM layer was washed with sat. NaHC03 (300 ml), followed by water (300 ml) and then with sat.brine solution (300 ml). The organic layer was dried over anhydrous Na2S04, and concentrated to afford the crude product, which upon column purification yielded the desired compound (10.0 g, yield: 23.0%). 1H NMR (DMSO-d6, 300 MHz): δ 3.23-3.21 (m, 9H), and 1.39 (s, 9H).
Step 2: Synthesis of tert-butyl 4-(methylsulfonyl)piperazine-l-carboxylate:
p
BocN N-S—
— ' II
o
Triethyl amine (22 ml, 160 mmol, 3.0 eq), methane sulphonyl chloride (4.5 ml, 59 mmol, 1.1 eq) followed by DMAP (0.66 g, 5.4 mmol, 0.1 eq) were added at 0 °C to a solution of tert-butyl piperazine- 1-carboxylate (step 1, 10.0 g, 54 mmol, 1.0 eq) in DCM (100 mL) and stirred for overnight at room temperature. After the completion of reaction (monitored by TLC), the reaction mass was washed with water (2x100 ml) followed by sat. brine solution (100 ml). The organic layer was dried over anhyd. Na2S04, and concentrated. The compound thus obtained was used directly for the next step without further purification (10.0 g, yield: 71.0%). 1H NMR (DMSO-d6, 300 MHz): δ 3.42 (t, 4H, J = 4.8 Hz), 3.06 (t, 4H, J = 4.8 Hz), 2.88 (s, 3H) and 1.40 (s, 9H).
Step 3: Synthesis of 1 -(methylsulfonyl)piperazine hydrochloride:
Figure imgf000044_0002
A solution of tert-butyl 4-(methylsulfonyl)piperazine- 1-carboxylate (step 2, 10 g, 38.0 mmol, 1.0 eq) in 1,4-dioxane.HCl (100 mL, 10 vol.) was stirred at room temperature for about 4 hours. After completion of the reaction (monitored by TLC), the volatile solvent was concentrated to afford the desired compound (6.0 g, yield: 80.0%) as a pale yellow solid, which was used directly for the next step without further purification 1H NMR (DMSO-d6, 300 MHz): δ 9.42 (bs, 2H), 3.35 (t, 4H, J = 5.8 Hz), 3.22-3.18 (m, 4H) and 2.99 (s, 3H).
Step 4: Synthesis of 1 -(2-methyl-2-nitropropyl)-4-(methylsulfonyl)piperazine:
Figure imgf000045_0001
2N aq. NaOH solution (12 ml) was added to l-(methylsulfonyl)piperazine hydrochloride (step 3, 4 g, 20 mmol, 1.0 eq) in a round bottomed flask. 30% formaldehyde (17 ml, 205 mmol, 10.2 eq) and 2-nitro propane (8.5 ml, 94 mmol, 4.7 eq) were added around 15-20 °C to the reaction mixture followed by addition of another lot of 2N aq. NaOH (12 ml) solution. The reaction mixture was stirred at room temperature for an hour after which was heated to 60 °C for about 3-4 hours. After completion of the reaction (monitored by TLC), the temperature was brought down to room temperature. The reaction mass was poured into ice cold water (50 ml) and the compound was extracted with ethylacetate (2x50 ml). The combined organic layers were washed with water (50 ml) followed by sat. brine solution (50 ml). The organic layer was dried over anhyd. Na2S04, and concentrated. The residue was purified by column chromatoghraphy to afford the desired pure compound (4.0 g, yield: 75.0%). 1H NMR (CDC13, 300 MHz): δ 3.03 (t, 4H, = 4.5 Hz), 2.88 (s, 2H), 2.85 (s, 3H), 2.55 (t, 4H, = 4.5 Hz) and 1.50 (s, 6H).
Step 5: Synthesis of 2-methyl-l -(4-(methylsulfonyl)piperazin-l -yl)propan-2-amine:
Raney Ni (10 mol%) was added to a solution of l-(2-methyl-2-nitropropyl)-4-(methyl sulfonyl)piperazine (step 4, 4.0 g, 15 mmol, 1.0 eq) in methanol (40 ml). The reaction mass was stirred for overnight at room temperature under hydrogen atmosphere. After completion of reaction (monitored by TLC), the reaction mass was filtered over celite and celite pad was washed with 10% MeOH in DCM (2x20 ml). The filtrate was dried over anhyd. Na2S04, and concentrated. The residue was purified by column chromatoghraphy to yield pure desired compound (0.79 g, yield: 20.0%). 1H NMR (CDC13, 300 MHz): δ 3.28 (bs, 2H), 3.09 (t, 4H, = 4.5 Hz), 2.87 (s, 3H), 2.60 (t, 4H, = 4.5 Hz), 2.21 (s, 2H) and 1.01 (s, 6H).
Intermediate 13: Preparation of (S)-l-ethyl-4-(pyrrolidin-2-ylmethyl)piperazine:
Figure imgf000045_0002
Step 1: Synthesis of (tert-butoxycarbony -L-proline:
Figure imgf000045_0003
L-proline (61.0 g, 472.86 mmol, 1.0 eq) dissolved in DCM (300 mL) which was stirred at 0 °C then added triethylamine (131.7 mL, 945.7 mmol, 3.0 eq) drop wise over 10 minutes and stirred at same temperature for about 30 minutes. Then added (Boc)20 (162.7 mL, 709.3 mmol, 1.5 eq) in DCM (300 mL) over 15 minutes which was stirred at room temperature for overnight. The reaction mixture was diluted with DCM and washed with water several times and brine solution. The organic layer was dried with anhydrous Na2S04, and concentrated to afford the desired compound (80.0 g, yield: 74.0%), which was proceed for next step without purification.
Step 2: Synthesis of tert-butyl (S)-2-(4-ethylpiperazine-l -carbonyl)pyrrolidine-l -carboxylate:
Figure imgf000046_0001
A solution of (tert-butoxycarbonyl)-L-proline (step 1, 25.0 g, 116.82 mmol, 1.0 eq) in DMF (250 mL) was treated sequentially with HOBt (18.9 g, 140.18 mmol, 1.2 eq) and EDCI (33.4 g, 170.5 mmol, 1.5 eq) and 1 -ethylpiperazine (14.67 g, 175.23 mmol, 1.5 eq). The suspension was stirred for about 15 minutes, TEA (80 mL, 584.0 mmol, 5.0 eq) was added, and the mixture stirred for about 12 hours. After completion of the reaction (monitored by TLC), the reaction was quenched by pouring into 50 mL of ethyl acetate, water and extracting with ethylacetate, washed with water, saturated aqueous NaHC03 and a portion of brine. The solution was then dried over Na2S04, filter and concentrated. The residue was purified by column chromatography on silica gel 2% MeOH and DCM to afford the desired product (5.5 g) as an off white solid. 1H NMR (DMSO-d6, 300 MHz): δ 4.70-4.55 (m, 1H), 3.57-3.38 (m, 4H), 2.44-2.26 (m, 6H), 2.17-2.16 (m, 2H), 1.80-1.67 (m, 4H), 1.39-1.31 (m, 9H) and 1.01 (t, 3H); Mass: [M-H]+ 312.19 (20%).
Step 3: Synthesis of (S)-l -ethyl-4-prolylpiperazine:
)
H O
To the stirred solution of tert-butyl (S)-2-(4-ethylpiperazine-l-carbonyl)pyrrolidine-l- carboxylate (step 2, 5.5 g, 17.7 mmol, 1.0 eq) in 1,4-dioxane.HCl (55 mL, 10 vol.) which was stirred at room temperature for about 4 hours. After completion of the reaction (monitored by TLC), the volatile solvent was concentrated to afford the hydrochloride salt compound as a pale yellow solid. Next reaction was carried out without any further purification.
Step 4: Synthesis of (S)-l -ethyl-4-(pyrrolidin-2-ylmethyl)piperazine :
LiAlH4 (1.9 g, 49.98 mmol, 3.0 eq) in THF (20 mL) was at 0 °C, then added drop wise (S)-l-ethyl-4-prolylpiperazine (step 3, 3.5 g, 16.6 mmol, 1.0 eq) in THF (15 mL) and the reaction mixture was stirred at room temperature for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by cautious addition of sodium sulphate past at 0 °C and stirring was continued for about 1 hour. The reaction mixture was filtered and the residue was washed with 10% MeOH and DCM. Then dried over anhydrous Na2S04 and concentrated organic solvent afford the desired product (3.0 g, yield: 92.0%) as a brown colour liquid. Next reaction was carried out without any further purification. 1H NMR (CDC13, 300 MHz): δ 3.70-3.64 (m, IH), 3.49-3.47 (m, IH), 3.39-3.36 (m, IH), 3.29-3.27 (m, IH), 3.19-3.14 (m, IH), 2.87-2.81 (m, IH), 2.73-2.70 (m, IH), 2.33- 2.19 (m, 9H), 2.10-2.09 (m, IH), 1.77-1.58 (m, 3H) and 0.99 (t, IH); Mass: [M-H]+ 198.16 (40%).
The below intermediates 14-27 were prepared by the procedure similar (including reagents and reaction conditions) to the above described in the synthesis of intermediate- 13 by using suitable reagents, reactents and solvents at appropriate conditions.
Figure imgf000047_0001
1H NMR (CDCI3, 300 MHz): δ 3.22 (m,
1H), 3.07-2.98 (m, 1H), 2.81-2.78 (m, 2H), 2.76-2.64 (m, 2H), 2.59 (m, 3H),
Intermediate 17
2.39-2.57 (m, 3H), 1.74-1.66 (m, 2H),
H and 1.58-1.52 (m, 8H); Mass: [M+H]+
183.12 (100%).
1H NMR (CDCI3, 300 MHz): δ 3.05- 3.01 (m, 1H), 2.81-2.76 (m, 1H), 2.71- 2.55 (m, 4H), 2.39-2.32 (m, 2H), 2.26-
Intermediate 18 2.18 (m, 2H), 2.07 (m, 6H), 1.79-1.61
(m, 2H), 1.59-1.53 (m, 4H), and 1.23-
H 1.18 (m, 1H); Mass: [M+H]+ 198.14
(100%).
1H NMR (CDCI3, 300 MHz): δ 3.70- 3.64 (m, 1H), 3.49-3.47 (m, 1H), 3.39- 3.36 (m, 1H), 3.29-3.27 (m, 1H), 3.19- 3.14 (m, 1H), 2.87-2.81 (m, 1H), 2.73-
Intermediate 19
2.70 (m, 1H), 2.33-2.19 (m, 9H), 2.10-
Figure imgf000048_0001
2.09 (m, 1H), 1.77-1.58 (m, 6H) and
0.99 (m, 3H); Mass: [M+H]+ 226.16 (40%).
1H NMR (CDCI3, 300 MHz): δ 3.48- 3.38 (m, 3H), 3.24-3.9 (m, 1H), 2.86- 2.77 (m, 3H), 2.27-2.19 (m, 2H), 1.91-
Intermediate 20
1.56 (m, 7H), 1.35-1.09 (m, 3H) and
H 0.80 (d, 3H); Mass: [M+H]+ 197.01
(100%).
1H NMR (CDCI3, 300 MHz): δ 3.64- 3.38 (m, 3H), 3.17-3.11 (m, 1H), 2.86- 2.80 (m, 1H), 2.75-2.67 (m, 1H), 2.50-
Intermediate 21 2.54 (m, 1H), 2.39 (m, 6H), 2.28-2.17
(m, 2H), 1.78-1.58 (m, 3H), 1.26-1.23
H
(m, 1H) and 0.95 (d, 6H); Mass: [M+H]+ 212.12 (100%).
Figure imgf000049_0001
Intermediate 28: Preparation of Synthesis of (R)-l-(pyrrolidin-2-ylmethyl)pyrrolidine:
Figure imgf000050_0001
Step 1: Synthesis of (tert-butoxycarbony -D-proline:
Figure imgf000050_0002
D-proline (61.0 g, 472.86 mmol, 1.0 eq) dissolved in DCM (300 mL) which was stirred at 0 °C then added triethylamine (131.7 mL, 945.7 mmol, 3.0 eq) drop wise over about 10 minutes and which was stirred at same temperature for about 30 minutes. After (Boc)20 (162.7 mL, 709.3 mmol, 1.5 eq) in DCM (300 mL) was added over about 15 minutes which was stirred at room temperature for overnight. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water several times and brain solution and organic layer was dried with anhydrous Na2S04, and concentrated under reduced pressure to afford the desired product (80.0 g, yield: 74.0%), which was proceed for the next step without purification.
Step 2: Synthesis of tert-butyl (R)-2-(pyrrolidine-l -carbonyl)pyrrolidine-l -carboxylate:
Figure imgf000050_0003
A solution of (tert-butoxycarbonyl)-D-proline (step 1, 5.g, 23.0 mmol, 1.0 eq) in DMF (25 mL) was treated sequentially with HOBt (6.0 g, 35.0 mmol, 1.5 eq) and EDCI ( 13.0 g, 69.0 mmol, 3.0 eq) and pyrrolidine (35.0 g, 46.0 mmol, 2.0 eq). The suspension was stirred for about 15 minutes, DIPEA (20 mL, 35.0 mmol, 1.5 eq) was added and the mixture stirred for 12 hours. The reaction was quenched by pouring into 50 mL of ethyl acetate and water extracting with ethylacetate and washed with water, saturated aqueous NaHC03, and a portion of brine. The solution was then dried over Na2S04, filtered and concentrated. The residue was purified by column chromatography on silica gel 2% MeOH and DCM to afford the desired product (3.5 g, yield: 59.0%) as an off white solid. 1H NMR (DMSO-d6, 300 MHz): δ 4.38-4.29 (m, IH), 3.51-3.23 (m, 6H), 2.19-1.90 (m, IH), 1.88-1.72 (m, 7H) and 0.69 (s, 9H); Mass: [M+Na]+ 291.16 (60%).
Step 3: Synthesis of (R)-l -prolylpyrrolidine :
Figure imgf000050_0004
To the stirred solution of tert-butyl (R)-2-(pyrrolidine-l-carbonyl)pyrrolidine-l- carboxylate (step 2, 3.5 g, 13.2 mmol, 1.0 eq) in 1,4-dioxane.HCl (30 mL) which was stirred at room temperature for about 4 hours. After completion of the reaction (monitored by TLC), the volatile solvent was concentrated to afford the desired compound as a brown liquid which was proceed for the next step without purification.
Step 4: Synthesis of (R)-l -(pyrrolidin-2-ylmethyl)pyrrolidine:
To a stirred solution of LiAlH4 (1.3 g, 36.46 mmol, 3.0 eq) in THF (70 mL) at 0 °C, was added drop wise (R)-l-prolylpyrrolidine (step 3, 2.2 g, 12.15 mmol, 1.0 eq) in THF (30 mL) and the reaction mixture was stirred at room temperature for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by cautious addition of sodium sulphate paste at 0 °C and stirring was continued for about 1 hour. The reaction mixture was filtered and the residue was washed with 10% MeOH and DCM, then dried over anhydrous Na2S04 and concentrated organic solvent to afford the desired compound (1.5 g, yield: 75.0%) as a brown colour liquid which was proceed for the next step without purification. 1H NMR (CDC13, 300 MHz): δ 3.10-2.97 (m, 1H), 2.82-2.63 (m, 2H), 2.53-2.46 (m, 2H), 2.24-2.17 (m, 1H), 2.08-1.99 (m, 3H), 1.83-1.53 (m, 6H), and 1.33-1.20 (m, 2H); Mass: [M+H]+ 155.14 (100%).
Intermediate 29: Preparation of (S)-2-((benzyloxy)methyl)-l-(((S)-pyrrolidin-2-yl)methyl) pyrrolidine hydrochloride:
Figure imgf000051_0001
Step 1: Synthesis of tert-butyl (S)-2-((S)-2-(hydroxymethyl)pyrrolidine-l-carbonyl) pyrrolidine-1 -carboxylate:
Figure imgf000051_0002
To a stirred solution of (tert-butoxycarbonyl)-L-proline (Intermediate 13-step 1, 7 g, 32.55 mmol) in DCM (70 ml) at 0 °C, were added HOBt (5.27 g, 39.06 mmol), EDCI (7.46 g, 39.06 mmol) and DIPEA (8.4 ml, 48.83 mmol). After stirring about 20 minutes a solution of (S)-pyrrolidin-2-ylmethanol (3.58 g, 35.81 mmol) in DCM (25 ml) was added and allowed to stir at room temperature for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, IN HC1 solution, saturated NaHC03 solution, brine and dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by column chromatography with EtOAc- hexane (7:3) as an eluent to afford the desired product (6.8 g, yield: 75.5%) as a thick oil. 1H MR (300 MHz, DMSO-d6): δ 4.74-4.68 (m, 1H), 4.38-4.29 (m, 2H), 3.94 (m, 1H), 3.54- 3.16 (m, 4H), 1.99-1.69 (m, 8H), 1.31 (s, 9H).
Step 2: Synthesis of tert-butyl (S)-2-(((S)-2-(hydroxymethyl)pyrrolidin-l-yl)methyl) pyrrolidine-1 -carboxylate:
Figure imgf000052_0001
To a stirred suspension of NaBH4 (2.58 g, 68.45 mmol) in dry THF (20 ml) at 0 °C, was added BF3:Et20 (7.1 ml, 57.04 mmol). After stirring about 30 minutes, was added a solution of tert-butyl (S)-2-((S)-2-(hydroxymethyl)pyrrolidine-l-carbonyl)pyrrolidine-l- carboxylate (step 1, 6.8 g, 22.81 mmol) in THF (60 ml) and allowed to stir at 60 °C for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by the cautious addition of MeOH (50 ml) at 0 °C and allowed to stir at 60 °C for about 12 hours. The reaction mixture was concentrated and the resulting residue was diluted with EtOAc and washed with water, brine and dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by column chromatography with MeOH:DCM (5:95) as an eluent to afford the desired product (3.6 g, yield: 55.6%) as a thick oil. 1H MR (300 MHz, CDC13): δ 3.96-3.84 (m, 1H), 3.61-3.54 (m, 1H), 3.40-3.04 (m, 4H), 2.68-2.19 (m, 4H), 1.88-1.68 (m, 8H), 1.47 (s, 9H).
Step 3: Synthesis of tert-butyl (S)-2-(((S)-2-((benzyloxy)methyl)pyrrolidin-l-yl)methyl) pyrrolidine-1 -carboxylate:
Figure imgf000052_0002
To a stirred solution of NaH (1.01 g, 25.35 mmol, 60% in mineral oil w/w) in THF (10 ml) under N2 atmosphere at 0 °C, was added tert-butyl (S)-2-(((S)-2-(hydroxymethyl) pyrrolidin-l-yl)methyl)pyrrolidine-l -carboxylate (step 2, 3.6 g, 12.67 mmol) in THF (40 ml). After 30 minutes benzyl bromide (2 ml, 16.47 mmol) was added and the reaction mixture was slowly allowed to attain to room temperature and stirred for overnight. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by addition of saturated NH4C1 solution at 0 °C. The solution was extracted with EtOAc (2x100 mL) and the combined organic phases were washed with brine, dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by column chromatography with MeOH:DCM (3:97) as an eluent to afford the desired product (3 g, yield: 63.29%) as an oil. 1H MR (300 MHz, CDC13): δ 7.37-7.29 (m, 5H), 4.51 (s, 2H), 3.91-3.78 (m, 1H), 3.45-3.19 (m, 5H), 2.66-2.59 (m, 2H), 2.46-2.37 (m, 1H), 2.28-2.18 (m, 1H), 1.96-1.55 (m, 8H), 1.46 (s, 9H).
Step 4: Synthesis of (S)-2-((benzyloxy)methyl)-l-(((S)-pyrrolidin-2-yl)methyl)pyrrolidine hydrochloride:
To a stirred solution of tert-butyl (S)-2-(((S)-2-((benzyloxy)methyl)pyrrolidin- l- yl)methyl)pyrrolidine-l-carboxylate (step 3, 1.5 g, 4.01 mmol) in dioxane (10 ml), was added 6N HC1 in dioxane (20 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (1.32 g, yield: 100%). Next reaction was carried out without any further purification.
Intermediate 30: Preparation of (R)-l-(((S)-pyrrolidin-2-yl)methyl)-3-(((R)-tetrahydrofuran- 3-yl)oxy)pyrrolidine:
Figure imgf000053_0001
Step 1: Synthesis of tert-butyl (R)-3-(((R)-tetrahydrofuran-3-yl)oxy)pyrrolidine-l- carboxylate:
Figure imgf000053_0002
To a stirred solution of NaH (1.38 g, 36.3 mmol, 60% in mineral oil w/w) in DMF (10 ml) under N2 atmosphere at 0 °C, was added tert-butyl (R)-3-hydroxypyrrolidine- l- carboxylate (Intermediate 8-step 1, 3.4 g, 18.0 mmol) in DMF (40 ml). After 30 minutes (S)- tetrahydrofuran-3-yl 4-methylbenzenesulfonate (4.84 g, 21.8 mmol) was added and the reaction mixture was slowly allowed to attain to room temperature and stirred for overnight. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by addition of saturated NH4C1 solution at 0 °C. The solution was extracted with EtOAc (2x100 mL) and the combined organic phases were washed with brine, dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by column chromatography with EtOAc:hexane (30:70) as an eluent to afford the desired product (0.9 g, yield: 28.1%) as an oil. 1H MR (300 MHz, DMSO-d6): δ 4.22-4.17 (m, 1H), 4.08-4.06 (m, 1H), 3.72-3.56 (m, 4H), 3.28-3.17 (m, 4H), 1.91 -1.82 (m, 4H), 1.39 (s, 9H).
Step 2: Synthesis of (R)-3-(((R)-tetrahydrofuran-3-yl)oxy)pyrrolidine hydrochloride:
Figure imgf000054_0001
To a stirred solution of tert-butyl (R)-3-(((R)-tetrahydrofuran-3-yl)oxy)pyrrolidine-l- carboxylate (step 1, 0.9 g, 3.50 mmol) in dioxane (5 ml), was added 6N HCl in dioxane (5 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (0.67 g, yield: 100%). Next reaction was carried out without any further purification.
Step 3: Synthesis of tert-butyl (S)-2-((R)-3-(((R)-tetrahydrofuran-3-yl)oxy)pyrrolidine-l- carbonyl )pyrrolidine-l -carboxylate:
Figure imgf000054_0002
To a stirred solution of (tert-butoxycarbonyl)-L-proline (Intermediate 13-step 1, 0.742 g, 3.4 mmol) and (R)-3-(((R)-tetrahydrofuran-3-yl)oxy)pyrrolidine hydrochloride (step 2, 0.67 g, 3.4 mmol) and DMAP (0.11 g, 0.9 mmol) in DCM (15 ml), was slowly added DCC (1.5 g, 7.2 mmol) in DCM (10 ml) at 0 °C and allowed to stir at room temperature for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHC03 solution, brine and dried over Na2S04. The solvent was evaporated and to the resulting solid, was added DCM (8 ml) and stirred for about 1 hour and filtered. The filtrate was concentrated under reduced pressure to afford the title compound (0.9 g, yield: 73%) as gummy oil. 1H MR (300 MHz, CDC13): δ 4.44-4.02 (m, 3H), 3.83-3.28 (m, 10H), 2.23-1.75 (m, 8H), 1.38 (s, 9H).
Step 4: Synthesis of (R)-l-(L-prolyl)-3-(((R)-tetrahydrofuran-3-yl)oxy)pyrrolidine hydrochloride:
Figure imgf000054_0003
To a stirred solution of tert-butyl (S)-2-((R)-3-(((R)-tetrahydrofuran-3-yl)oxy) pyrrolidine- l-carbonyl)pyrrolidine-l -carboxylate (step 3, 0.9 g, 2.52 mmol) in dioxane (5 ml), was added 6N HCl in dioxane (6 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (0.73 g, yield: 100%). Next reaction was carried out without any further purification.
Step 5: Synthesis of (R)-l-(((S)-pyrrolidin-2-yl)methyl)-3-(((R)-tetrahydrofuran-3-yl)oxy) pyrrolidine:
To a stirred suspension of LiAlH4 (0.16 g, 4.0 mmol) in dry THF (3 ml), was slowly added a solution of (R)-l-(L-prolyl)-3-(((R)-tetrahydrofuran-3-yl)oxy)pyrrolidine hydrochloride (step 4, 0.7 g, 2 mmol) in THF (10 ml) at 0 °C. After addition, the reaction mixture was then brought to room temperature and stirred for overnight. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by the sequential addition of 0.2 mL of H20, 0.6 mL of 15% aq. NaOH and 0.6 mL of H20. The mixture was then poured into EtOAc (30 ml) and stirred for about 30 minutes. The insoluble material was removed by filtration through celite. The filtrate was dried over Na2S04 and concentrated under reduced pressure to afford the desired product (0.6 g). Next reaction was carried out without any further purification.
Intermediate 31: Preparation of (S)-4,4-difluoro-l-(pyrrolidin-2-ylmethyl)piperidine:
Figure imgf000055_0001
Step 1: Synthesis of tert-butyl 4,4-difluoropi eridine-l -carboxylate :
Figure imgf000055_0002
To a stirred solution of tert-butyl 4-oxopiperidine-l -carboxylate (Intermediate 9-step 1 , 5.0 g, 25.12 mmol) in DCM (50 ml) under N2 atmosphere at 0 °C, was added DAST (3.7 ml, 27.63 mmol) and stirred for about 6 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by addition of ice cold water. The solution was extracted with DCM (2x100 mL) and the combined organic phases were washed with brine, dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by column chromatography with EtOAc and hexane (1 :9) to afford the desired compound (3.73 g, yield: 67.2%) as an oil. 1H MR (300 MHz, CDC13): δ 3.56-3.52 (m, 4H), 1.99-1.86 (m, 4H), 1.46 (s, 9H).
Step 2: Synthesis of 4,4-difluoropiperidine hydrochloride:
Figure imgf000056_0001
To a stirred solution of tert-butyl 4,4-difluoropiperidine- 1-carboxylate (step 1, 3.73 g, 16.87 mmol) in dioxane (20 ml), was added 6N HCl in dioxane (30 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (2.64 g, yield: 100%). Next reaction was carried out without any further purification.
Step 3: Synthesis of tert-butyl (S)-2-(4,4-difluoropiperidine-l-carbonyl)pyrrolidine-l- carboxylate:
Figure imgf000056_0002
To a stirred solution of (tert-butoxycarbonyl)-L-proline (Intermediate 13-step 1, 5.42 g, 25.22 mmol) and 4,4-difluoropiperidine hydrochloride (step 2, 2.64 g, 16.81 mmol) and DMAP (1.0 g, 8.4 mmol) in DCM (70 ml), was slowly added DCC (6.92 g, 33.63 mmol) in DCM (40 ml) at 0 °C and allowed to stir at room temperature for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM, washed with water, saturated NaHC03 solution, brine and dried over Na2S04. The solvent was evaporated and to the resulting solid, was added DCM (30 ml) and stirred for about one hour and filtered. The filtrate was concentrated under reduced pressure to afford the title compound (4.4 g, yield: 82.24%) as a solid. 1H MR (300 MHz, CDC13): δ 4.72-4.67 (m, 1H), 3.93-3.41 (m, 6H), 2.22-1.82 (m, 6H), 1.46 (s, 9H), 1.45-1.28 (m, 2H); ES Mass: 336.28 [M+ H4]+.
Step 4: Synthesis of (S)-4,4-difluoro-l -prolylpiperidine hydrochloride:
Figure imgf000056_0003
To a stirred solution of tert-butyl (S)-2-(4,4-difluoropiperidine-l-carbonyl) pyrrolidine- 1-carboxylate (step 3, 4.4 g, 13.82 mmol) in dioxane (30 ml), was added 6N HCl in dioxane (25 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (3.51 g, yield: 100%). Next reaction was carried out without any further purification. Step 5: Synthesis of (S)-4,4-difluoro-l -(pyrrolidin-2-ylmethyl)piperidine: To a stirred suspension of LiAlH4 (1.1 g, 27.63 mmol) in dry THF (15 ml), was slowly added a solution of (S)-4,4-difluoro-l-prolylpiperidine hydrochloride (step 4, 3.51 g, 13.81 mmol) in THF (40 ml) at 0 °C. After addition, the reaction mixture was then brought to room temperature and stirred for overnight. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by the sequential addition of 1.0 mL of H20, 3 mL of 15% aq. NaOH and 3 mL of H20. The mixture was then poured into EtOAc (100 ml) and stirred for about 30 minutes. The insoluble material was removed by filtration through celite and the solvent was removed from the filtrate by rotary evaporation and dried over Na2S04. The product was isolated by column chromatography on silica gel using MeOH: DCM (3:97) as an eluent to give the desired product (2.64 g, yield: 94%) as a solid. 1H NMR (300 MHz, DMSO-d6): δ 3.16-3.05 (m, 2H), 2.84-2.61 (m, 3H), 2.52 (m, 1H), 2.33-2.18 (m, 2H), 2.01- 1.47 (m, 6H), 1.39-1.01 (m, 2H); ES Mass: 205.15 [M+H]+.
Intermediate 32: Preparation of (S)-3,3-difluoro-l- rrolidin-2-ylmethyl)pyrrolidine:
Figure imgf000057_0001
Step 1: Synthesis of tert-butyl 3-oxopyrrolidine-l-carboxylate:
Figure imgf000057_0002
To a stirred solution of tert-butyl (R)-3-hydroxypyrrolidine-l-carboxylate (Intermediate 8-step 1, 4.5 g, 24.1 mmol) in DCM (60 ml) under N2 atmosphere, was added DMP (20.45 g, 48.12 mmol) at 0 °C. The reaction mixture was allowed to room temperature and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with 1 : 1 mixture of saturated NaHC03 and saturated Na2S203 solution. The reaction mixture was extracted with DCM (2x100 ml). The combined organic layer was washed with brine, dried over anhydrous sodium sulphate and concentrated under reduced pressure to give the residue. Purification by column chromatography with EtOAc and hexane (15:85) to afford the desired compound (3.44 g, yield: 77.3%) as an oil. 1H NMR (300 MHz, CDC13): δ 3.80-3.75 (m, 4H), 2.61-2.56 (m, 2H), 1.48 (s, 9H).
Step 2: Synthesis of tert-butyl 3,3-difluoropyrrolidine-l-carboxylate:
Figure imgf000057_0003
To a stirred solution of tert-butyl 3-oxopyrrolidine-l-carboxylate (step 1, 3.44 g, 18.59 mmol) in DCM (40 ml) under N2 atmosphere at 0 °C, was added DAST (2.7 ml, 20.45 mmol) and stirred for about 6 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by addition of ice cold water. The solution was extracted with DCM (2x100 mL) and the combined organic phases were washed with brine, dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by column chromatography with EtOAc and hexane (1 :9) to afford the desired compound (1.61 g, yield: 41.92%) as an oil. 1H MR (300 MHz, CDC13): δ 3.65-3.55 (m, 4H), 2.38-2.24 (m, 2H), 1.46 (s, 9H).
Step 3: Synthesis of 3,3-difluoropyrrolidine hydrochloride:
Figure imgf000058_0001
To a stirred solution of tert-butyl 3,3-difluoropyrrolidine-l-carboxylate (step 2, 1.61 g, 7.77 mmol) in dioxane (15 ml), was added 6N HCl in dioxane (20 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (1.11 g, yield: 100%). Next reaction was carried out without any further purification.
Step 4: Synthesis of tert-butyl (S)-2-(3,3-difluoropyrrolidine-l-carbonyl)pyrrolidine-l- carboxylate:
Figure imgf000058_0002
To a stirred solution of (tert-butoxycarbonyl)-L-proline (Intermediate 13-step 1, 2.5 g, 11.66 mmol) and 3,3-difluoropyrrolidine hydrochloride (step 3, 1.11 g, 7.77 mmol) and DMAP (0.47 g, 3.88 mmol) in DCM (30 ml), was slowly added DCC (3.2 g, 15.52 mmol) in DCM (20 ml) at 0 °C and allowed to stir at room temperature for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHC03 solution, brine and dried over Na2S04. The solvent was evaporated and to the resulting solid, was added DCM (15 ml) and stirred for about 1 hour and filtered. The filtrate was concentrated under reduced pressure to afford the title compound (2.35 g, yield: 100%) as a solid.
Step 5: Synthesis of (S)-3 ' ,3-difluoro-l-prolylpyrrolidine hydrochloride:
Figure imgf000059_0001
To a stirred solution of tert-butyl (S)-2-(3,3-difluoropyrrolidine-l-carbonyl) pyrrolidine- 1-carboxylate (step 4, 2.35 g, 7.73 mmol) in dioxane (20 ml), was added 6N HCl in dioxane (30 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (2.13 g, yield: 100%). Next reaction was carried out without any further purification. Step 6: Synthesis of (S)-3,3-difluoro-l-(pyrrolidin-2-ylmethyl)pyrrolidine:
To a stirred suspension of LiAlH4 (0.58 g, 15.43 mmol) in dry THF (10 ml), was slowly added a solution of (S)-3,3-difluoro-l-prolylpyrrolidine hydrochloride (step 5, 2.13 g, 7.71 mmol) in THF (25 ml) at 0 °C. After addition, the reaction mixture was then brought to room temperature and stirred for overnight. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by the sequential addition of 0.6 mL of H20, 1.8 mL of 15% aq. NaOH and 1.8 mL of H20. The mixture was then poured into EtOAc (100 ml) and stirred for about 30 minutes. The insoluble material was removed by filtration through celite and the solvent was removed from the filtrate by rotary evaporation, dried over Na2S04. The product was isolated by column chromatography on silica gel using MeOH: DCM (2:98) as an eluent to give the desired product (1.43 g, yield: 98%) as a solid. 1H NMR (300 MHz, DMSO-d6): δ 3.38-3.27 (m, 3H), 3.07-3.03 (m, 1H), 2.92-2.67 (m, 4H), 2.36-2.34 (m, 1H), 2.26-2.14 (m, 2H), 1.76-1.58 (m, 4H); ES Mass: 191.10 [M+H]+.
Intermediate 33: Preparation of (S)-3-fluoro-l-(((S)-pyrrolidin-2-yl)methyl)pyrrolidine dihydrochloride:
Figure imgf000059_0002
Step 1: Synthesis of tert-butyl (S)-2-(((S)-3-fluoropyrrolidin-l-yl)methyl)pyrrolidine-l- carboxylate:
Figure imgf000059_0003
To a stirred solution of tert-butyl (S)-2-(((R)-3-hydroxypyrrolidin-l-yl)methyl) pyrrolidine- 1-carboxylate (1.0 g, 3.7 mmol) in DCM (15 ml) under N2 atmosphere at 0 °C, was added DAST (0.6 ml, 4.0 mmol) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by addition ice cold water. The solution was extracted with DCM (2x20 mL) and the combined organic phases were washed with brine, dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by column chromatography with MeOH and DCM (1 :99) to afford the desired compound (0.5 g, yield: 50%) as an oil.
Step 2: Synthesis of (S)-3-fluoro-l-(((S)-pyrrolidin-2-yl)methyl)pyrrolidine dihydrochloride:
To a stirred solution of tert-butyl (S)-2-((4-fluoropiperidin-l-yl)methyl)pyrrolidine-l- carboxylate (step 1, 0.5 g, 1.8 mmol) in dioxane (5 ml), was added 6N HCl in dioxane (5 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (0.444 g, yield: 100%). Next reaction was carried out without any further purification.
Intermediate 34: Preparation of (2S,4R)-2-(pyrrolidin-l-ylmethyl)-4-(((R)-tetrahydrofuran-3- yl)oxy)pyrrolidine hydrochloride:
Figure imgf000060_0001
Step 1: Synthesis of tert-butyl (2S,4R)-4-hydroxy-2-(pyrrolidine-l-carbonyl)pyrrolidine-l- carboxylate:
Figure imgf000060_0002
To a stirred solution of (2S,4R)-l-(tert-butoxycarbonyl)-4-hydroxypyrrolidine-2- carboxylic acid (10 g, 43.27 mmol) in DCM (120 ml) at 0 °C, were added HOBt (7 g, 51.92 mmol), EDCI (9.9 g, 51.92 mmol) and DIPEA (11.23 ml, 64.9 mmol). After stirring for about 20 minutes pyrrolidine (3.9 ml, 47.5 mmol) was added and allowed to stir at room temperature for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, IN HCl solution, saturated NaHC03 solution, brine, dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by column chromatography with EtOAc-hexane (1: 1) as an eluent to afford the desired product (8 g, yield: 65.5%) as a thick oil. 1H MR (300 MHz, DMSO-d6): δ 4.99 (d, J = 3.3 Hz, 1H), 4.45-4.38 (m, 1H), 4.26 (m, 1H), 3.56-3.22 (m, 6H), 2.11-2.02 (m, 1H), 1.91-1.69 (m, 5H), 1.30 (s, 9H). Step 2: Synthesis of tert-butyl (2S,4R)-4-hydroxy-2-(pyrrolidin-l-ylmethyl)pyrrolidine-l- carboxylate:
Figure imgf000061_0001
To a stirred suspension of NaBH4 (3.2 g, 84.47 mmol) in dry THF (30 ml) at 0 °C, was added BF3:Et20 (8.6 ml, 70.39 mmol). After stirring for about 30 minutes, was added a solution of tert-butyl (2S,4R)-4-hydroxy-2-(pyrrolidine-l-carbonyl)pyrrolidine-l-carboxylate (step 1, 8 g, 28.15 mmol) in THF (50 ml) and allowed to stir at 60 °C for about 2 hours. After completion of the reaction (monitored by TLC), reaction mixture was quenched by the cautious addition of MeOH (70 ml) at 0 °C and allowed to stir at 60 °C for about 12 hours. The reaction mixture was concentrated and the resulting residue was diluted with EtOAc and washed with water, brine, dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by column chromatography with MeOH:DCM (5:95) as an eluent to afford the desired product (4.8 g, yield: 63.15%) as a thick oil. 1H NMR (300 MHz,
DMSO-d6): δ 4.84 (d, J = 3.3 Hz, 1H), 4.21-4.16 (m, 1H), 3.82 (m, 1H), 3.23-3.20 (m, 3H), 2.51-2.40 (m, 5H), 1.90-1.87 (m, 2H), 1.66-1.62 (m, 4H), 1.39 (s, 9H); ES Mass: 271.29
[M+H]+.
Step 3: Synthesis of tert-butyl (2S,4R)-2-(pyrrolidin-l-ylmethyl)-4-(((R)-tetrahydrofuran-3-yl) oxy )pyrrolidine-l -carboxylate:
Figure imgf000061_0002
To a stirred solution of NaH (1.86 g, 46.66 mmol, 60% in mineral oil w/w) in DMF (10 ml) under N2 atmosphere at 0 °C, was added tert-butyl (2S,4R)-4-hydroxy-2-(pyrrolidin- l-ylmethyl)pyrrolidine-l -carboxylate (step 2, 1.8 g, 6.66 mmol) in DMF (15 ml). After 30 minutes (S)-tetrahydrofuran-3-yl 4-methylbenzenesulfonate (1.6 g, 6.66 mmol) was added and the reaction mixture was slowly allowed to attain to room temperature and stirred for overnight. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by addition of saturated NH4C1 solution at 0 °C. The solution was extracted with EtOAc (2x50 mL) and the combined organic phases were washed with brine, dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by column chromatography with MeOH:DCM (5:95) as an eluent to afford the desired product (0.38 g, yield: 16.8%) as an oil. 1H NMR (300 MHz, CDC13): δ 4.23-4.20 (m, 1H), 4.14-4.08 (m, 2H), 3.92-3.41 (m, 5H), 2.75-2.49 (m, 7H), 2.19-1.71 (m, 8H), 1.46 (s, 9H); ES Mass:
341.30 [M+H]+.
Step 4: Synthesis of (2S,4R)-2-(pyrrolidin-l-ylmethyl)-4-(((R)-tetrahydrofuran-3-yl)oxy) pyrrolidine hydrochloride:
To a stirred solution of tert-butyl (2S,4R)-2-(pyrrolidin-l-ylmethyl)-4-(((R)-tetra hydrofuran-3-yl)oxy)pyrrolidine-l-carboxylate (step 3, 0.38 g, 1.11 mmol) in dioxane (2 ml), was added 6N HC1 in dioxane (4 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (3.4 g, yield: 100%). Next reaction was carried out without any further purification.
Intermediate 35: Preparation of 2-((S)-l-(((S)-pyrrolidin-2-yl)methyl)pyrrolidin-2-yl)propan- 2-ol:
Figure imgf000062_0001
Step 1: Synthesis of tert-butyl (S)-2-(2-hydroxypropan-2-yl)pyrrolidine-l-carboxylate:
Figure imgf000062_0002
To a stirred solution of Mg (3.14 g, 261.66 mmol) in Et20 (40 ml), were added I2 (cat) and iodomethane (9 ml, 130.8 mmol) and stirred for about 30 minutes. Then a solution of 1- (tert-butyl) 2-methyl (S)-pyrrolidine-l,2-dicarboxylate (6 g, 26.2 mmol) in THF (60 ml) was added and stirred for overnight. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by cautious addition of saturated NH4C1 solution at 0 °C and extracted with EtOAc (2x100 ml). The combined organic layers were washed with brine, dried over anhydrous sodium sulphate and concentrated under reduced pressure to get the residue. The crude product was purified via silica gel column chromatography with EtOAc and n-hexane (30:70) as an eluent to afford the desired compound (5.9 g, yield: 98.3%) as a thick oil. 1H MR (300 MHz, DMSO-d6): δ 4.84 (brs, 1H), 3.70-3.68 (m, 1H), 3.51-3.47 (m, 1H), 3.12 (m, 1H), 1.81-1.61 (m, 4H), 1.39 (s, 9H), 1.04 (s, 6H); ES Mass: 252.16 [M+Na]+. Step 2: Synthesis of (S)-2-(pyrrolidin-2-yl)propan-2-ol hydrochloride:
Figure imgf000062_0003
To a stirred solution of tert-butyl (S)-2-(2-hydroxypropan-2-yl)pyrrolidine-l- carboxylate (step 1, 5.9 g, 25.7 mmol) in dioxane (50 ml), was added 6N HC1 in dioxane (50 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (4.2 g, yield: 100%). Next reaction was carried out without any further purification.
Step 3: Synthesis of tert-butyl (S)-2-((S)-2-(2-hydroxypropan-2-yl)pyrrolidine-l-carbonyl) pyrrolidine-1 -carboxylate:
Figure imgf000063_0001
To a stirred solution of (tert-butoxycarbonyl)-L-proline (Intermediate 13 -step 1, 8.2 g, 38.01 mmol) and (S)-2-(pyrrolidin-2-yl)propan-2-ol hydrochloride (step 2, 4.2 g, 25.3 mmol) and DMAP (1.54 g, 12.4 mmol) in DCM (120 ml), was slowly added DCC (10.4 g, 50.7 mmol) in DCM (60 ml) at 0 °C and allowed to stir at room temperature for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM, washed with water, saturated NaHC03 solution, brine and dried over Na2S04. The solvent was evaporated and to the resulting solid, was added DCM (90 ml) and stirred for about one hour and filtered. The filtrate was concentrated under reduced pressure to afford the title compound (8.0 g, yield: 97.5%) as gummy oil. 1H MR (300 MHz, DMSO-d6): δ 6.15 (s, 1H), 4.63-4.60 (m, 1H), 4.26-1.19 (m, 1H), 4.02-3.95 (m, 1H), 3.50-3.29 (m, 3H), 2.12-2.02 (m, 3H), 1.98-1.89 (m, 3H), 1.72-1.68 (m, 2H), 1.41 (s, 9H), 1.19 (s, 3H), 1.03 (s, 3H); ES Mass: 349.30 [M+Na]+.
Step 4: Synthesis of (S)-l-(L-prolyl)-2- -hydroxypropan-2-yl)pyrrolidine hydrochloride:
Figure imgf000063_0002
To a stirred solution of tert-butyl (S)-2-((S)-2-(2-hydroxypropan-2-yl)pyrrolidine-l- carbonyl)pyrrolidine- 1 -carboxylate (step 3, 8.0 g, 24.5 mmol) in dioxane (50 ml), was added 6N HC1 in dioxane (80 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (6.4 g, yield: 100%). Next reaction was carried out without any further purification.
Step 5: Synthesis of2-((S)-l-(((S)-pyrrolidin-2-yl)methyl)pyrrolidin-2-yl)propan-2-ol:
To a stirred suspension of LiAlH4 (1.63 g, 42.8 mmol) in dry THF (20 ml), was slowly added a solution of (S)-l-(L-prolyl)-2-(2-hydroxypropan-2-yl)pyrrolidine hydrochloride (step 4, 6.4 g, 21.4 mmol) in THF (50 ml) at 0 °C. After addition, the reaction mixture was then brought to room temperature and stirred for overnight. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by the sequential addition of 2.0 mL of H20, 6 mL of 15 % aq. NaOH and 6 mL of H20. The mixture was then poured into EtOAc (100 ml) and stirred for about 30 minutes. The insoluble material was removed by filtration through celite and the solvent was removed from the filtrate by rotary evaporation and dried over Na2S04. The product was isolated by flash column chromatography on silica gel using MeOH: DCM (5:95) as an eluent to give the desired product (4.4 g, yield: 98%) as a thick oil. 1H MR (300 MHz, CDC13): δ 3.43-3.25 (m, 3H), 3.08-2.92 (m, 3H), 2.76-2.55 (m, 2H), 1.85-1.43 (m, 8H), 1.11 (s, 3H), 1.10 (s, 3H); ES Mass: 213.62 [M+H]+.
Intermediate 36: Preparation of (S)-2-(pyrrolidin-l-ylmethyl)-l-(pyrrolidin-2-ylmethyl)-lH- pyrrole hydrochloride:
Figure imgf000064_0001
Step 1: Synthesis of tert-butyl (S)-2-(hydroxymethyl)pyrrolidine-l -carboxylate :
Figure imgf000064_0002
To a suspension of lithium aluminium hydride (17.6 g, 0.46 mol, 2.0 eq) in THF (500 ml) at 0 °C, (tert-butoxycarbonyl)-L-proline (Intermediate 13-step 1, 50 g, 0.23 mol, 1.0 eq) in THF (500 ml) was added dropwise over a period of about 1 hour. The reaction was allowed to stir at room temperature for about 90 minutes. The reaction mass was quenched at 0 °C using sat. sodium sulphate, followed by addition of ethylacetate. The reaction mass was filtered over celite. The organic layer from the filtrate was separated and compound was extracted from water layer with ethylacetate. Combined organic layers were dried over anhydrous Na2S04 and concentrated under reduced pressure. The crude compound was purified by column chromatographic technique to afford the desired compound (24 g, yield: 51%), which was used for the next step without further purification. 1H NMR (DMSO, 300 MHz): δ 4.67 (t, 1H, = 5.5 Hz), 3.65 (bs, 1H), 3.50-3.43 (m, 1H), 3.26-3.18 (m, 3H), 1.82- 1.70 (m, 4H), 1.39 (s, 9H).
Step 2: Synthesis of tert-butyl ( S)-2-formylpy rrolidine-1 -carboxylate :
Figure imgf000064_0003
To a stirred solution of tert-butyl (S)-2-(hydroxymethyl)pyrrolidine-l-carboxylate (step 1, 2.5 g, 12.4 mmol) in DCM (30 ml) under N2 atmosphere, was added DMP (7.9 g, 18.6 mmol) at 0 °C. The reaction mixture was allowed to room temperature and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with 1 : 1 mixture of saturated NaHC03 and saturated Na2S203 solution. The reaction mixture was extracted with DCM (2x50 ml). The combined organic layer was washed with brine, dried over anhydrous sodium sulphate and concentrated under reduced pressure to give the residue (2.1 g, 84%). Next reaction was carried out without any further purification.
Step 3: Synthesis of tert-butyl (S)-2-(((S)-2-(hydroxymethyl)pyrrolidin-l-yl)methyl) pyrrolidine-1 -carboxylate:
Figure imgf000065_0001
To a stirred solution of tert-butyl (S)-2-formylpyrrolidine-l-carboxylate (step 2, 2.1 g, 10.5 mmol) in 1,2-dichloro ethane (25 ml) under N2 atmosphere, was added AcOH (1 ml, 15.8 mmol) at 0 °C. After stirring about 20 minutes at 0 °C, were added (S)-prolinol (1.1 g, 10.5 mmol) and followed by sodiumtriacetoxy borohydride (3.4 g, 15.8 mmol). The reaction mixture was allowed to room temperature and stirred for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with saturated NH4C1 solution. The reaction mixture was extracted with DCM (2x50 ml). The combined organic layers were washed with brine, dried over anhydrous sodium sulphate and concentrated under reduced pressure to give the residue. The residue was purified by silica gel column (elution 2% MeOH in DCM) to afford the title compound (1.6 g, yield: 55.1%) as an off white solid. 1H NMR (300 MHz, CDC13): δ 3.97-3.32 (m, 6H), 2.73-2.36 (m, 4H), 1.90-1.79 (m, 8H), 1.43 (s, 9H).
Step 4: Synthesis of tert-butyl (S)-2-((2-formyl-lH-pyrrol-l-yl)methyl)pyrrolidine-l- carboxylate:
Figure imgf000065_0002
To a stirred solution of tert-butyl (S)-2-(((S)-2-(hydroxymethyl)pyrrolidin-l-yl) methyl)pyrrolidine- 1 -carboxylate (step 3, 1.6 g, 5.6 mmol) in DCM (20 ml) under N2 atmosphere, was added DMP (3.6 g, 8.0 mmol) at 0 °C. The reaction mixture was allowed to room temperature and stirred for about 3 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with 1 : 1 mixture of saturated NaHC03 and saturated Na2S203 solution. The reaction mixture was extracted with DCM (2x30 ml). The combined organic layers were washed with brine, dried over anhydrous sodium sulphate and concentrated under reduced pressure to give the residue (1.4 g, yield: 93%). Next reaction was carried out without any further purification.
Step 5: Synthesis of tert-butyl (S)-2-((2-(pyrrolidin-l-ylmethyl)-lH-pyrrol-l-yl)methyl) pyrrolidine-1 -carboxylate:
Figure imgf000066_0001
To a stirred solution of tert-butyl (S)-2-((2-formyl-lH-pyrrol-l-yl)methyl)pyrrolidine- 1-carboxylate (step 4, 1.4 g, 4.0 mmol) in 1,2-dichloro ethane (20 ml) under N2 atmosphere, was added AcOH (0.5 ml, 7.4 mmol) at 0 °C. After stirring about 20 minutes at 0 °C, were added pyrrolidine (0.5 ml, 5.4 mmol) and followed by sodiumtriacetoxy borohydride (1.6 g, 7.4 mmol). The reaction mixture was allowed to room temperature and stirred for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with saturated NH4C1 solution. The reaction mixture was extracted with DCM (2x30 ml). The combined organic layers were washed with brine, dried over anhydrous sodium sulphate and concentrated under reduced pressure to give the residue. The residue was purified by silica gel column (elution 2% MeOH in DCM) to afford the title compound (1.1 g, yield: 78.5%) as an off white solid.1H NMR (300 MHz, CDC13): δ 6.63 (m, 1H), 6.13- 6.06 (m, 2H), 4.06-3.96 (m, 2H), 3.76-3.69 (m, 1H), 3.33-3.28 (m, 2H), 2.75-2.63 (m, 6H), 1.81-1.69 (m, 8H), 1.48 (s, 9H).
Step 6: Synthesis of (S)-2-(pyrrolidin-l-ylmethyl)-l-(pyrrolidin-2-ylmethyl)-lH-pyrrole hydrochloride:
To a stirred solution of tert-butyl (S)-2-((2-(pyrrolidin-l-ylmethyl)-lH-pyrrol-l-yl) methyl)pyrrolidine- 1 -carboxylate (step 5, 1.1 g, 3.3 mmol) in 1,4-dioxane (10 ml), was added 6N HC1 in 1,4-dioxane (10 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (0.88 g, yield: 100%). Next reaction was carried out without any further purification.
Intermediate 37: Preparation of (S)-4-((benzyloxy)methyl)-l-(pyrrolidin-2-ylmethyl) piperidine:
Figure imgf000066_0002
Step 1: Synthesis of tert-butyl (S)-2-(4-(hydroxymethyl)piperidine-l-carbonyl)pyrrolidine-l- carboxylate:
Figure imgf000067_0001
A solution of (tert-butoxycarbonyl)-L-proline (Intermediate 13-step 1, 5.0 g, 23.25 mmol, 1.0 eq) in DMF (50 mL) was treated sequentially with HOBt (5.3 g, 34.5 mmol, 1.5 eq) and EDCI (13.0 g, 69.00 mmol, 3.0 eq) and piperidin-4-ylmethanol (2.97 g, 25.58 mmol, 1.1 eq). The suspension was stirred for about 15 minutes, DIPEA (10.0 mL, 34.8 mmol, 5.0 eq) was added, and the mixture stirred for about 12 hours. After completion of the reaction (monitored by TLC), the reaction was quenched by pouring into 50 mL of ethyl acetate and water, extracting with ethylacetate, washed with water and a portion of brine. The solution was then dried over Na2S04, filter and concentrated. The residue was purified by column chromatography on silica gel 3% MeOH and DCM to afford the title compound (2.9 g, yield: 40.0%) as a brown colour liquid. 1H NMR (DMSO-d6, 300 MHz): δ 4.60 (m, 2H), 4.45-4.27 (m, 3H), 3.95-3.88 (m, IH), 3.30-3.24 (m, 2H), 3.03-2.91 (m, IH), 2.18 (m, IH), 1.84-1.69 (m, 6H), 1.28 (m, 9H) and 1.13-0.95 (m, 3H); Mass: [M+H]+ 213.05 (20%).
Step 2: Synthesis of tert-butyl (S)-2-(4-((benzyloxy)methyl)piperidine-l-carbonyl)pyrrolidine- 1-carboxylate:
Figure imgf000067_0002
A solution of tert-butyl (S)-2-(4-(hydroxymethyl)piperidine-l-carbonyl)pyrrolidine-l- carboxylate (step 1, 2.9 g, 9.2 mmol, 1.0 eq) in DMF (30 mL) then the reaction mixture was cooled at 0 °C and added sodium hydride (0.88 g, 37.06 mmol, 4.0 eq) followed benzylbromide (1.24 mL, 10.18 mmol, 1.1 eq). The reaction mixture was stirred for about 12 hours at room remparature. After completion of the reaction (monitored by TLC), the reaction was quenched by pouring into 50 mL of cold water, extracting with ethylacetate, washed with water and a portion of brine. The organic layer was dried over Na2S04, filter, and concentrated. The residue was purified by column chromatography on silica gel 5% MeOH and DCM to afford the title compound (2.8 g, yield: 75.0%) as a brown colour liquid. 1H NMR (DMSO-d6, 300 MHz): δ 7.36-7.30 (m, 5H), 4.60 (m, 2H), 4.45-4.27 (m, 3H), 3.95- 3.88 (m, 1H), 3.30-3.24 (m, 3H), 3.03-2.91 (m, 1H), 2.18 (m, 1H), 1.84-1.69 (m, 6H), 1.28 (m, 9H) and 1.13-0.95 (m, 3H); Mass: [M+H]+ 403.26 (100%).
Step 3: Synthesis of (S)-4-((benzyloxy)methyl)-l-prolylpiperidine hydrochloride:
Figure imgf000068_0001
To the stirred solution of tert-butyl (S)-2-(4-((benzyloxy)methyl)piperidine-l- carbonyl)pyrrolidine-l-carboxylate (step 2, 2.8 g, 6.9 mmol, 1.0 eq) in 1,4-dioxane.HCl (30 mL, 10 vol.) which was stirred at room temperature for about 4 hours. After completion of the reaction (monitored by TLC), the volatile solvent was concentrated to afford the title compound as a pale yellow solid, which was directly used in the next step without any further purification.
Step 4: Synthesis of (S)-4-((benzyloxy)methyl)-l-(pyrrolidin-2-ylmethyl)piperidine:
LiAlH4 (0.83 g, 19.80 mmol, 3.0 eq) in THF (20 mL) was at 0 °C, then added drop wise (S)-4-((benzyloxy)methyl)-l-prolylpiperidine hydrochloride (step 3, 2.0 g, 6.6 mmol, 1.0 eq) in THF (30 mL) and the reaction mixture was stirred at room temperature for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by cautious addition of sodium sulphate past at 0 °C and stirring was continued for about 1 hour. The reaction mixture was filtered, the residue was washed with 10% MeOH and DCM, then dried over anhydrous Na2S04 and concentrated organic solvent to afford the title compound (1.6 g, yield: 84.0%) as a brown colour liquid, which was directly used in the next step. 1H NMR (DMSO-d6, 300 MHz): δ 7.36-7.29 (m, 5H), 4.53 (m, 2H), 3.64-3.23 (m, 2H), 3.11-3.07 (m, 3H), 2.88-2.65 (m, 3H), 2.25-1.80 (m, 4H), 1.72-1.52 (m, 4H), 1.35-1.00 (m, 4H) and 0.88 (m, 1H); Mass: [M+H]+ 289.19 (100%).
Intermediate 38: Preparation of (2S,4S)-4-fluoro-2-((2-methoxyethoxy)methyl)-l-(((S)- p yrrolidin-2- vDmeth yl)p yrrolidine :
Figure imgf000068_0002
Step 1: Synthesis of tert-butyl (2S,4R)-4-hydroxy-2-(hydroxymethyl)pyrrolidine-l- carboxylate: HO
Boc
To a stirred suspension of LiAlH4 (2.95 g, 77.63 mmol) in dry THF (30 ml), was slowly added a solution of (2S,4R)-l-(tert-butoxycarbonyl)-4-hydroxypyrrolidine-2- carboxylic acid (12.0 g, 51.92 mmol) in THF (150 ml) at 0 °C. After addition, the reaction mixture was then brought to room temperature and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by the sequential addition of 3 mL of H20, 9 mL of 15% aq. NaOH and 9 mL of H20. The mixture was then poured into EtOAc (150 ml) and stirred for about 30 minutes. The insoluble material was removed by filtration through celite, the solvent was removed from the filtrate by rotary evaporation and dried over Na2S04. The product was isolated by flash column chromatography on silica gel using EtOAc: hexane (4:6) as an eluent to give the desired product (4.8 g, yield: 42.62%) as a thick oil. 1H NMR (300 MHz, DMSO-d6): δ 4.84-4.81 (m, 1H), 4.68-4.62 (m, 1H), 4.23-4.17 (m, 1H), 3.74 (m, 1H), 3.49-3.39 (m, 2H), 3.23-3.17 (m, 2H), 1.99-1.79 (m, 2H), 1.38 (s, 9H).
Step 2: Synthesis of tert-butyl (2S,4R)-4-hydroxy-2-((2-methoxyethoxy)methyl)pyrrolidine-l- carboxylate:
Figure imgf000069_0001
To a stirred solution of NaH (4.97 g, 124.42 mmol, 60 % in mineral oil w/w) in DMF
(50 ml) under N2 atmosphere at 0 °C, was added tert-butyl (2S,4R)-4-hydroxy-2- (hydroxymethyl)pyrrolidine-l-carboxylate (step 1, 9.0 g, 41.47 mmol) in THF (80 ml). After 30 minutes l-bromo-2-methoxyethane (5.9 ml, 62.21 mmol) was added and the reaction mixture was slowly allowed to attain room temperature and stirred for overnight. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by addition of saturated NH4C1 solution at 0 °C. The solution was extracted with EtOAc (2x250 mL) and the combined organic phases were washed with brine, dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by column chromatography with EtOAc-hexane (3:7) as an eluent to afford the desired product (4.9 g, yield: 43%) as an oil. 1H NMR (300 MHz, DMSO-d6): δ 4.88 (d, J = 2.1 Hz, 1H), 4.21 (m,
1H), 3.84 (m, 1H), 3.52-3.41 (m, 6H), 3.27-3.19 (m, 2H), 3.23 (s, 3H), 1.86 (m, 2H), 1.39 (s, 9H). Step 3: Synthesis of tert-butyl (2S,4S)-4-fluoro-2-((2-methoxyethoxy)methyl)pyrrolidine-l- carboxylate:
Figure imgf000070_0001
To a stirred solution of tert-butyl (2S,4R)-4-hydroxy-2-((2-methoxyethoxy)methyl) pyrrolidine- l-carboxylate (step 2, 1.4 g, 5.1 mmol) in DCM (15 ml) under N2 atmosphere at 0 °C, was added DAST (1.02 ml, 7.6 mmol) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by addition ice cold water. The solution was extracted with DCM (2x20 mL) and the combined organic phases were washed with brine, dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by column chromatography with EtOAc-hexane (2:8) to afford the desired compound (0.9 g, yield: 67.37%) as an oil. 1H MR (300 MHz, DMSO- d6): δ 5.31-5.14 (m, 1H), 3.93 (m, 1H), 3.68-3.61 (m, 1H), 3.52-3.42 (m, 6H), 3.26-3.17 (m, 1H), 3.23 (s, 3H), 2.19-1.97 (m, 2H), 1.41 (s, 9H).
Step 4: Synthesis of (2S,4S)-4-fluor -2-((2-methoxyethoxy)methyl)pyrrolidine hydrochloride:
Figure imgf000070_0002
To a stirred solution of tert-butyl (2S,4S)-4-fluoro-2-((2-methoxyethoxy)methyl) pyrrolidine- l-carboxylate (step 3, 0.95 g, 3.43 mmol) in dioxane (10 ml), was added 6N HCl in dioxane (5 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (0.73 g, yield: 100%). Next reaction was carried out without any further purification. Step 5: Synthesis of tert-butyl (S)-2-((2S,4S)-4-fluoro-2-((2-methoxyethoxy)methyl) pyrrolidine-1 -carbonyl )pyrrolidin -l -carboxylate:
Figure imgf000070_0003
To a stirred solution of (tert-butoxycarbonyl)-L-proline (Intermediate 13-step 1, 0.543 g, 2.52 mmol), (2S,4S)-4-fluoro-2-((2-methoxyethoxy)methyl)pyrrolidine hydrochloride (step 4, 0.73 g, 2.52 mmol), TEA (0.7 ml, 5.05 mmol) and DMAP (0.06 g, 0.5 mmol) in DCM (15 ml), was slowly added DCC (1.1 g, 5.05 mmol) in DCM (10 ml) at 0 °C and allowed to stir at room temperature for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHC03 solution, brine and dried over Na2S04. The solvent was evaporated and to the resulting solid, was added DCM (7 ml) and stirred for about 1 hour and filtered. The filtrate was concentrated under reduced pressure to afford the title compound (1.0 g, yield: 78.12%) as a gummy oil. 1H MR (300 MHz, DMSO-d6): δ 5.4-5.2 (m, 1H), 4.46-4.42 (m, 1H), 4.22-4.13 (m, 1H), 3.63-3.20 (m, 10H), 3.22 (s, 3H), 2.20-2.04 (m, 2H), 1.79-1.67 (m, 4H), 1.29 (s, 9H).
Step 6: Synthesis of (2S,4S)-l-(L-prolyl)-4-fluoro-2-((2-methoxyethoxy)methyl)pyrrolidine hydrochloride:
Figure imgf000071_0001
To a stirred solution of tert-butyl (S)-2-((2S,4S)-4-fluoro-2-((2-methoxyethoxy) methyl)pyrrolidine-l-carbonyl)pyrrolidine-l-carboxylate (step 5, 1.0 g, 2.67 mmol) in dioxane (10 ml), was added 6N HCl in dioxane (6 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (0.83 g, yield: 100%). Next reaction was carried out without any further purification.
Step 7: Synthesis of (2S,4S)-4-fluoro-2-((2-methoxyethoxy)methyl)-l-(((S)-pyrrolidin-2-yl) methyl pyrrolidine :
To a stirred suspension of LiAlH4 (0.17 g, 4.27 mmol) in dry THF (3 ml), was slowly added a solution of (2S,4S)-l-(L-prolyl)-4-fluoro-2-((2-methoxyethoxy)methyl)pyrrolidine hydrochloride (step 6, 0.83 g, 2.14 mmol) in THF (10 ml) at 0 °C. After addition, the reaction mixture was then brought to room temperature and stirred for overnight. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by the sequential addition of 0.2 mL of H20, 0.6 mL of 15% aq. NaOH and 0.6 mL of H20. The mixture was then poured into EtOAc (30 ml) and stirred for about 30 minutes. The insoluble material was removed by filtration through celite. The filtrate was dried over Na2S04 and concentrated under reduced pressure to afford the desired product (0.5 g, yield: 72.46%). Next reaction was carried out without any further purification.
Intermediate 39: Preparation of (S)-l-(pyrrolidin-2-ylmethyl)pyrrolidine hydrochloride:
Figure imgf000071_0002
Step 1: Synthesis of tert-butyl (S)-2-((tosyloxy)methyl)pyrrolidine-l-carboxylate:
Figure imgf000072_0001
Triethyl amine (0.8 ml, 5.9 mmol, 1.2 eq), followed by p-toluenesulphonyl chloride (1.04 g, 5.5 mmol, 1.1 eq) and catalytic amount of DMAP were added at 0 °C to a solution of tert-butyl (S)-2-(hydroxymethyl)pyrrolidine-l-carboxylate (Intermediate 36-step 1, 1 g, 4.9 mmol, 1.0 eq) in DCM (10 ml). The reaction was stirred for about 4 hours at room temperature. The reaction mass was diluted with water (10 ml) and the organic layer was separated. The compound was further extracted with DCM (2x10 ml). Combined organic layers were dried over anhydrous Na2S04 and concentrated under reduced pressure. The crude compound was purified by column chromatographic technique to afford the desired compound (1.1 g, yield: 63%), which was used for the next step without further purification. 1H NMR (CDC13, 300 MHz): δ 7.77 (d, 2H, = 8.1 Hz), 7.35 (d, 2H, = 8.1 Hz), 4.08 (d, 2H, = 4.8 Hz), 3.96-3.89 (m, 2H), 3.29 (d, 2H, = 6.3 Hz), 2.45 (s, 3H), 1.93-1.80 (m, 4H), 3.26-3.18 (m, 3H), 1.38 (s, 9H).
Step 2: Synthesis of tert-butyl (S)-2-(pyrrolidin-l -ylmethyl)pyrrolidine-l -carboxylate:
Figure imgf000072_0002
Pyrrolidine (0.28 ml, 3.4 mmol, 1.1 eq) was added to a solution of tert-butyl (S)-2- ((tosyloxy)methyl)pyrrolidine-l -carboxylate (step 1, 1.1 g, 3.1 mmol, 1.0 eq) in toluene (10 ml) and refluxed for overnight. The reaction mass was washed with saturated Na2C03 solution, followed by water and brine solution. The organic layer was dried over anhydrous Na2S04 and concentrated under reduced pressure. The crude compound was purified by column chromatographic technique to afford the desired compound (0.4 g, yield: 51%), which was used for the next step without further purification. 1H NMR (DMSO, 300 MHz): δ 3.76 (bs, 1H,), 3.21-3.19 (m, 2H), 2.43-2.41 (m, 5H), 1.82-1.66 (m, 8H), 1.39 (s, 9H); MS(ESI): m/z 255.15 (M + H)+.
Step 3: Synthesis of (S)-l -(pyrrolidin-2-ylmethyl)pyrrolidine hydrochloride:
Tert-butyl (S)-2-(pyrrolidin-l-ylmethyl)pyrrolidine-l -carboxylate (step 2) was dissolved in a solution of HCl in dioxane (4 mL) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the solvent was evaporated under reduced pressure to give title compound, which was directly used in the next step.
Intermediate 40: Preparation of (3'S,5'S)-5'-(pyrrolidin-l-ylmethyl)-l,3'-bipyrrolidine hydrochloride:
Figure imgf000073_0001
Step 1: Synthesis of tert-butyl (2S,4R)-4-((methylsulfonyl)oxy)-2-(((methylsulfonyl)oxy) methyl)pyrrolidine-l-carboxylate:
Figure imgf000073_0002
To a stirred solution of tert-butyl (2S,4R)-4-hydroxy-2-(hydroxymethyl)pyrrolidine-l- carboxylate (Intermediate 38-step 1, 2 g, 9.21 mmol) in DCM (25 ml) under N2 atmosphere, were added N(Et)3 (5.2 ml, 36.8 mmol), followed by MsCl (1.8 ml, 22.2 mmol) and catalytic amount of DMAP at 0 °C. The reaction mixture was allowed to room temperature and stirred for about 3 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM. The organic layer was washed with saturated H4C1 solution, brine, dried over anhydrous sodium sulphate and concentrated under reduced pressure to get the residue (3.43 g) as thick oil. Next reaction was carried out without any further purification. Step 2: Synthesis of tert-butyl (3'S,5'S)-5'-(pyrrolidin-l-ylmethyl)-[l,3'-bipyrrolidine]-l '- carboxylate:
Figure imgf000073_0003
To a stirred solution of tert-butyl (2S,4R)-4-((methylsulfonyl)oxy)-2-(((methyl sulfonyl)oxy)methyl)pyrrolidine-l-carboxylate (step 1, 3.4 g, 9.11 mmol) in ACN (50 ml) under N2 atmosphere, were added Cs2C03 (14.8 g, 45.5 mmol), followed by pyrrolidine (3.8 ml, 45.5 mmol) at room temperature. The reaction mixture was heated to 100 °C and continued for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to room temperature and filtered through celite and the cake was washed with DCM (twice). The filtrate was washed with water, brine, dried over anhydrous sodium sulphate and concentrated under reduced pressure to give the residue. The crude product was purified via silica gel column chromatography with MeOH and DCM (5:95) to afford the desired compound (1.0 g, yield: 34.12%) as thick oil. 1H NMR (300 MHz, DMSO- d6): δ 3.78-3.74 (m, 1H), 3.57 (m, 1H), 2.97-2.91 (m, 1H), 2.61-2.56 (m, 3H), 2.42 (m, 8H), 2.18-2.08 (m, 1H), 1.81-1.76 (m, 1H), 1.66 (m, 8H), 1.39 (s, 9H).
Step 3: Synthesis of '(3'S,5'S)-5 '-(pyrrolidin-1 -ylmethyl)-! ,3 ' -bipyrrolidine hydrochloride: To a stirred solution of tert-butyl (3'S,5'S)-5'-(pyrrolidin-l-ylmethyl)-[l,3'- bipyrrolidine]-l '-carboxylate (step 2, 1.0 g, 3.1 mmol) in dioxane (5 ml), was added 6N HCl in dioxane (10 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (0.91 g). Next reaction was carried out without any further purification.
Intermediate 41: Preparation of (S)-4-fluoro-l-(pyrrolidin-2-ylmethyl)piperidine dihydrochloride:
Figure imgf000074_0001
Step 1: Synthesis of tert-butyl (S)-2-((4-hydroxypiperidin-l-yl)methyl)pyrrolidine-l- carboxylate:
Figure imgf000074_0002
To a stirred solution of tert-butyl (S)-2-((tosyloxy)methyl)pyrrolidine-l-carboxylate (Intermediate 39-step 1, 4.0 g, 10.95 mmol) in 1,4-dioxane (50 ml) under N2 atmosphere, were added Cs2C03 (10.7 g, 32.87 mmol), followed by piperidin-4-ol (1.1 g, 10.95 mmol) at room temperature. The reaction mixture was heated to 100 °C and continued for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to room temperature and filtered through celite and the cake was washed with DCM (twice). The filtrate was washed with water, brine, dried over anhydrous sodium sulphate and concentrated under reduced pressure to give the residue. The crude product was purified via silica gel column chromatography with MeOH and DCM (3 :97) to afford the desired compound (1.7 g, yield: 55.1%) as an off white solid. 1H NMR (300 MHz, DMSO-d6): δ 4.46 (d, J = 3.6 Hz, 1H), 3.73 (m, 1H), 3.40 (m, 1H), 3.19-3.16 (m, 2H), 2.81-2.77 (m, 1H), 2.62- 2.58 (m, 1H), 2.30-2.26 (m, 1H), 2.16-2.09 (m, 2H), 1.99-1.92 (m, 1H), 1.80-1.62 (m, 6H), 1.38 (s, 9H), 1.35-1.23 (m, 2H).
Step 2: Synthesis of tert-butyl (S)-2-((4-fluoropiperidin-l-yl)methyl)pyrrolidine-l- carboxylate:
Figure imgf000074_0003
To a stirred solution of tert-butyl (S)-2-((4-hydroxypiperidin-l-yl)methyl)pyrrolidine-
1-carboxylate (step 1, 1.7 g, 5.98 mmol) in DCM (25 ml) under N2 atmosphere at 0 °C, was added DAST (1.1 ml, 6.58 mmol) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by addition ice cold water. The solution was extracted with DCM (2x50 mL) and the combined organic phases were washed with brine, dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by column chromatography with MeOH and DCM (2:98) to afford the desired compound (0.5 g, yield: 29.4%) as an oil. 1H MR (300 MHz, CDC13): δ 4.76-4.60 (m, 1H), 4.28-4.16 (m, 1H), 4.0-3.82 (m, 1H), 3.35-3.31 (m, 2H), 2.88-2.23 (m, 5H), 2.05-1.85 (m, 6H), 1.46 (s, 9H), 1.27 (m, 2H).
Step 3: Synthesis of (S)-4-fluoro-l -(pyrrolidin-2-ylmethyl)piperidine dihydrochloride:
To a stirred solution of tert-butyl (S)-2-((4-fluoropiperidin-l-yl)methyl)pyrrolidine-l- carboxylate (step 2, 0.5 g, 1.74 mmol) in dioxane (5 ml), was added 6N HC1 in dioxane (5 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (0.452 g, yield: 100%). Next reaction was carried out without any further purification.
Intermediate 42: Preparation of (lR,2S,5S)-6,6-dimethyl-2-(pyrrolidin-l-ylmethyl)-3- azabicyclor3.1.Olhexane dihydrochloride:
Figure imgf000075_0001
Step 1: Synthesis of 3 -(tert-butyl) 2-methyl (lR,2S,5S)-6,6-dimethyl-3-az bicyclo[3.1.0] hexane-2,3-dicarboxylate:
Figure imgf000075_0002
To a stirred solution of methyl (lR,2S,5S)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2- carboxylate hydrochloride (1 g, 4.8 mmol) in 1,4 dioxane and water (8:2 ml) at 0 °C, were added 2N NaOH (2 ml) and (Boc)20 (1.3 ml, 5.8 mmol). The reaction mixture was slowly allowed to attain to room temperature and stirred for overnight. After completion of the reaction (monitored by TLC), the reaction mixture was extracted with DCM (2x20 mL) and the combined organic phases were washed with brine, dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by column chromatography with EtOAc-hexane (1: 9) as an eluent to afford the desired product (1.3 g, yield: 99%) as an oil. 1H NMR (300 MHz, DMSO-d6): δ 3.97 (s, 1H), 3.68 (s, 3H), 3.53-3.47 (m, 1H), 3.32- 3.25 (m, 1H), 1.43-1.39 (m, 2H), 1.30 (s, 9H), 1.00 (s, 3H), 0.91 (s, 3H).
Step 2: Synthesis of tert-butyl (lR,2S,5S)-2-(hydroxymethyl)-6,6-dimethyl-3-azabicyclo[3.1.0] hexane-3-carboxylate:
Figure imgf000076_0001
To a stirred suspension of LiAlH4 (0.23 g, 5.8 mmol) in dry THF (5 ml), was slowly added a solution of 3 -(tert-butyl) 2-methyl (lR,2S,5S)-6,6-dimethyl-3-azabicyclo[3.1.0] hexane-2,3-dicarboxylate (step 1, 1.3 g, 4.83 mmol) in THF (15 ml) at 0 °C. After addition, the reaction mixture was then brought to room temperature and stirred for about 1 hour. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by the sequential addition of 0.3 mL of H20, 0.9 mL of 15% aq. NaOH and 0.9 mL of H20. The mixture was then poured into EtOAc (30 ml) and stirred for about 30 minutes. The insoluble material was removed by filtration through celite and the solvent was removed from the filtrate by rotary evaporation and dried over Na2S04. The product was isolated by flash column chromatography on silica gel using EtOAc: hexane (2:8) as an eluent to give the desired product (1.16 g, yield: 99%) as a thick oil. 1H NMR (300 MHz, DMSO-d6): δ 4.79- 4.72 (m, 1H), 3.54-3.46 (m, 2H), 3.39-3.24 (m, 3H), 1.36 (s, 9H), 1.32 (m, 1H), 1.26-1.24 (m, 1H), 0.99 (s, 3H), 0.82 (s, 3H).
Step 3: Synthesis of tert-butyl (lR,2S,5S)-6,6-dimethyl-2-(pyrrolidin-l-ylmethyl)-3-azabicyclo [3.1.0]hexane-3-carboxylate:
Figure imgf000076_0002
To a stirred solution of tert-butyl (lR,2S,5S)-2-(hydroxymethyl)-6,6-dimethyl-3- azabicyclo[3.1.0]hexane-3-carboxylate (step 2, 1.16 g, 4.8 mmol) in DCM (15 ml) under N2 atmosphere, were added N(Et)3 (1.4 ml, 9.6 mmol), followed by MsCl (0.5 ml, 5.8 mmol) at 0 °C. The reaction mixture was allowed to room temperature and stirred for about 1 hour. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM. The organic layer was washed with saturated NH4C1 solution, brine, dried over anhydrous sodium sulphate and concentrated under reduced pressure to give tert-butyl (lR,2S,5S)-6,6-dimethyl-2-(((methylsulfonyl)oxy)methyl)-3-azabicyclo[3.1.0]hexane-3- carboxylate (1.3 g). Next reaction was carried out without any further purification. To a stirred solution of tert-butyl (lR,2S,5S)-6,6-dimethyl-2-(((methylsulfonyl)oxy) methyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (1.3 g, 4.1 mmol) in ACN (20 ml) under N2 atmosphere, were added Cs2C03 (2.0 g, 6.11 mmol), followed by pyrrolidine (0.4 ml, 4.9 mmol) at room temperature. The reaction mixture was heated to 80 °C and continued for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to room temperature and filtered through celite and the cake was washed with EtOAc (twice). The filtrate was washed with water, brine, dried over anhydrous sodium sulphate and concentrated under reduced pressure to give the residue. The crude product was purified via silica gel column chromatography with MeOH and DCM (2:98) to afford the desired compound (1.19 g, yield: 99%) as a thick oil. 1H NMR (300 MHz, DMSO-d6): δ 3.58 (ddd, J = 3.6, 9.3, 18.9 Hz, 1H), 3.38-3.25 (m, 1H), 3.28 (s, 2H), 2.58-2.48 (m, 2H), 2.44- 2.33 (m, 3H), 1.67-1.66 (m, 4H), 1.41-1.39 (m, 1H), 1.36 (s, 9H), 1.25-1.22 (m, 1H), 0.98 (s, 3H), 0.83 (s, 3H).
Step 4: Synthesis of (lR,2S,5S)-6,6-dimethyl-2-(pyrrolidin-l-ylmethyl)-3-az bicyclo[3.1.0] hexane dihydrochloride:
To a stirred solution of tert-butyl (lR,2S,5S)-6,6-dimethyl-2-(pyrrolidin-l-ylmethyl)- 3-azabicyclo[3.1.0]hexane-3-carboxylate (step 3, 1.19 g, 4.1 mmol) in dioxane (10 ml), was added 6N HC1 in dioxane (10 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (1.07 g, yield: 100%). Next reaction was carried out without any further purification.
Intermediate 43: Preparation of l-((2S)-2-((lH-pyrazol-l-yl)methyl)-l 4-pyrrolidin-l-yl)- 2,2,2-trifluoroethan- 1 -one:
Figure imgf000077_0001
Synthesis of tert-butyl (S)-2-((lH-pyrazol-l-yl)methyl)pyrrolidine-l-carboxyL
Figure imgf000077_0002
Pyrazole (0.43 g, 6.3 mmol, 1.5 eq) was added to a solution of Cs2C03 (3.16 g, 9.69 mmol, 2.3 eq) and tert-butyl (S)-2-((tosyloxy)methyl)pyrrolidine-l-carboxylate (Intermediate 39-step 1, 1.5 g, 4.22 mmol, 1.0 eq) in DMF (30 ml) stirred for overnight. After completion of the reaction (monitored by TLC), the solvent was evaporated under reduced pressure, followed by water wash and brine solution. The organic layer was dried over anhydrous Na2S04 and concentrated under reduced pressure. The crude compound was purified by column chromatographic technique to afford the desired compound (0.68 g, yield: 64%), which was used for the next step without further purification. 1H NMR (CDC13, 300 MHz): δ 1.36-1.39 (m, 1H), 1.50 (s, 9H), 1.87-1.93 (m, 2H), 3.13-3.36 (m, 2H), 4.02-4.09 (m, 1H), 4.20-4.35 (m, 2H), 6.25 (s, 1H), 7.31-7.35 (m, 1H), 7.48 (s, 1H); MS(ESI): m/z 252.16 (M + H)+.
Step 2: Synthesis of l-((2S)-2-((lH-pyrazol-l-yl)methyl)-lX4-pyrrolidin-l-yl)-2,2,2-trifluoro ethan-l-one:
TFA (1.0 ml, 8.7 mmol, 5 eq) was added to a solution of tert-butyl (S)-2-((lH- pyrazol-l-yl)methyl)pyrrolidine-l-carboxylate (step 1, 680 mg, 2.7 mmol, 1 eq) in DCM (15 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the solvent was evaporated under reduced pressure to give title compound, which was directly used in the next step.
Intermediate 44: Preparation of (R)-3-(2-methoxyethoxy)-l-(((S)-pyrrolidin-2-yl)methyl) pyrrolidine dihydrochloride:
Figure imgf000078_0001
Step 1: Synthesis of tert-butyl (S)-2-(((R)-3-hydroxypyrrolidin-l-yl)methyl)pyrrolidine-l- carboxylate:
Figure imgf000078_0002
To a stirred solution of tert-butyl (S)-2-((tosyloxy)methyl)pyrrolidine-l-carboxylate
(Intermediate 39-step 1, 6.0 g, 16.8 mmol) in ACN (50 ml) under N2 atmosphere, were added Cs2C03 (16.4 g, 50.6 mmol), followed by (R)-pyrrolidin-3-ol hydrochloride (2.2 g, 16.8 mmol) at room temperature. The reaction mixture was heated to 100 °C and continued for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to room temperature and filtered through celite and the cake was washed with DCM (twice). The filtrate was washed with water, brine, dried over anhydrous sodium sulphate and concentrated under reduced pressure to give the residue. The crude product was purified via silica gel column chromatography with MeOH and DCM (2:98) to afford the desired compound (1.8 g, yield: 40.8%) as an off white solid. Step 2: Synthesis of tert-butyl (S)-2-(((R)-3-(2-methoxyethoxy)pyrrolidin-l-yl)methyl) pyrrolidine-1 -carboxylate:
Figure imgf000079_0001
To a stirred solution of NaH (1.0 g, 26.6 mmol, 60% in mineral oil w/w) in DMF (7 ml) under N2 atmosphere at 0 °C, was added tert-butyl (S)-2-(((R)-3-hydroxypyrrolidin-l- yl)methyl)pyrrolidine-l -carboxylate (step 1, 1.8 g, 6.6 mmol) in DMF (15 ml). After 30 minutes l-bromo-2-methoxyethane (1.0 ml, 9.9 mmol) was added and the reaction mixture was slowly allowed to attain to room temperature and stirred for overnight. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by addition of saturated NH4C1 solution at 0 °C. The solution was extracted with EtOAc (2x75 mL) and the combined organic phases were washed with brine, dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by column chromatography with EtOAc-hexane (2: 8) as an eluent to afford the desired product (1.5 g, yield: 69%) as an oil. 1H MR (300 MHz, CDC13): δ 4.06-4.02 (m, 1H), 3.92-3.82 (m, 1H), 3.53 (m, 4H), 3.37 (s, 3H), 3.32 (m, 2H), 2.86-2.47 (m, 6H), 2.14-1.80 (m, 6H), 1.46 (s, 9H); ESI MS: 329.3 (M+H)+.
Step 3: Synthesis of (R)-3-(2-methoxyethoxy)-l-(((S)-pyrrolidin-2-yl)methyl)pyrrolidine dihydrochloride:
To a stirred solution of tert-butyl (S)-2-(((R)-3-(2-methoxyethoxy)pyrrolidin-l- yl)methyl)pyrrolidine-l -carboxylate (step 2, 1.5 g, 4.5 mmol) in dioxane (10 ml), was added 6N HC1 in dioxane (15 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (1.38 g, yield: 100%). Next reaction was carried out without any further purification.
The below intermediate 45 were prepared by the procedure similar (including reagents and reaction conditions) to the above described in the synthesis of intermediate-44 by using suitable reagents, reactents and solvents at appropriate conditions.
Figure imgf000079_0002
Intermediate 46: Preparation of l-((2S)-2-((3,5-dimethyl-lH-pyrazol-l-yl)methyl)-l 4- pyrrolidin-l-yl)-2,2,2-trifluoroethan-l-one:
Figure imgf000080_0001
Step 1: Synthesis of tert-butyl (S)-2-((3,5-dimethyl-lH-pyrazol-l-yl)methyl)pyrrolidine-l- carboxylate:
Figure imgf000080_0002
To a stirred solution of 3,5-dimethyl pyrazole (2 g, 20.83 mmol, 1.0 eq) in DMF (20 ml) added sodium hydride (1.0 g, 41.66 mmol, 2 eq) at 0 °C and stirred for about 30 minutes at ambient temperature, then added tert-butyl (S)-2-((tosyloxy)methyl)pyrrolidine-l- carboxylate (Intermediate 39-step 1, 7.39 g, 20.83 mmol, 1 eq) in DMF (30 ml) and stirred for about 2 hours at 80 °C. After completion of the reaction (monitored by TLC), the reaction mass was quenched with ice cold water after that extracted with ethyl acetate, followed by water wash and brine solution. The organic layer was dried over anhydrous Na2S04 and concentrated under reduced pressure. The crude compound was purified by column chromatographic technique to afford the desired compound (3.0 g, yield: 51.7%), which was used for the next step without further purification. 1H NMR (CDC13, 300 MHz): δ 1.47 (s, 9H), 1.73-1.78 (m, 4H), 2.19-2.28 (m, 5H), 3.11-3.61 (m, 3H), 3.91-4.23 (m, 3H), 5.78 (s, 1H)); MS(ESI): m/z 280.21 (M + H)+.
Step 2: Synthesis of l-((2S)-2-((3,5-dimethyl-lH-pyrazol-l-yl)methyl)-lX4-pyrrolidin-l-yl)- 2,2,2-trifluoroethan-l-one:
TFA (4.48 ml, 53.76 mmol, 5 eq) was added to a solution of tert-butyl (S)-2-((3,5- dimethyl-lH-pyrazol-l-yl)methyl)pyrrolidine-l-carboxylate (step 1, 3 g, 10.75 mmol, 1 eq) in DCM (15 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the solvent was evaporated under reduced pressure to afford the title compound, which was directly used in the next step.
The below intermediates 47-51 were prepared by the procedure similar (including reagents and reaction conditions) to the above described in the synthesis of intermediate-46 by using suitable reagents, reactents and solvents at appropriate conditions.
Intermediate no Structure
Figure imgf000081_0001
Intermediate 52: Preparation of l-(p rrolidin-l-ylmethyl)cyclobutan-l -amine hydro chloride:
Figure imgf000081_0002
Step 1: Synthesis of methyl 1 -((tert-butoxycarbonyl)amino)cyclobutane-l -carboxylate:
Figure imgf000081_0003
To a stirred solution of 1-aminocyclobutane-l-carboxylic acid (2.7 g, 23.4 mmol) in MeOH (40 ml), was carefully added acetyl chloride (4.2 ml, 58.6 mmol) drop wise at 0 °C. The reaction mixture was then warmed to room temperature and stirred for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to give methyl 1-aminocyclobutane-l -carboxylate hydrochloride (3.02 g, yield: 100 %) as a white solid.
To a stirred solution of methyl 1-aminocyclobutane-l -carboxylate hydrochloride (3.02 g, 23.4 mmol) in THF (40 ml), were added N(Et)3 (9.6 ml, 69.6 mmol) and (Boc)20 (6.4 ml, 23.8 mmol) at 0 °C. The reaction mixture was then warmed to room temperature and stirred for about 12 hours. After completion of the reaction (monitored by TLC), saturated NH4C1 solution was added, extracted with EtOAc (2 X 100 mL). The combined organic phases were washed with brine, dried over Na2S04 and concentrated under reduced pressure to give the crude product. Purification by flash chromatography with EtOAc-hexane (3: 7) as an eluent to afford the desired product (5.32 g, yield: 100%) as an oil. 1H NMR (300 MHz, DMSO-d6): δ 7.64 (s, 1H), 3.60 (s, 3H), 2.44-2.38 (m, 2H), 2.14-2.04 (m, 2H), 1.88-1.80 (m, 2H), 1.36 (s, 9H).
Step 2: Synthesis of tert-butyl (l-(hydrox methyl)cyclobutyl)carbamate:
Figure imgf000082_0001
To a stirred suspension of LiAlH4 (1.8 g, 46.4 mmol) in dry THF (15 ml), was slowly added a solution of methyl l-((tert-butoxycarbonyl)amino)cyclobutane-l-carboxylate (step 1, 5.32g, 23.2 mmol) in THF (15 ml) at 0 °C. The reaction mixture was brought to room temperature and stirred for about 3 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched by the sequential addition of 1.8 mL of H20, 5.4 mL of 15% aq. NaOH and 5.4 mL of H20. The mixture was then poured into EtOAc (100 ml) and stirred for about 30 minutes. The reaction mixture was filtered through celite, dried over Na2S04 and concentrated under reduced pressure. The product was isolated by flash column chromatography on silica gel using (EtOAc: hexane-70:30) to give the desired product (3.7 g, yield: 80%) as an oil. 1H NMR (300 MHz, DMSO-d6): δ 6.62 (s, 1H), 4.67 (t, J = 5.4 Hz, 1H), 3.41 (d, J = 5.4 Hz, 2H), 2.22-2.09 (m, 2H), 2.01-1.90 (m, 2H), 1.65-1.57 (m, 2H), 1.36 (s, 9H).
Step 3: Synthesis of (l-((tert-butoxycarbonyl)amino)cyclobutyl)methyl 4-methylbenzene sulfonate:
Figure imgf000082_0002
To a stirred solution of tert-butyl (l-(hydroxymethyl)cyclobutyl)carbamate (step 2, 3.7 g, 18.4 mmol) in DCM (40 ml) under N2 atmosphere, were added N(Et)3 (7.8 ml, 55.2 mmol), followed by TsCl (6 g, 31.3 mmol) and catalytic amount of DMAP at 0 °C. The reaction mixture 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. The organic layer was washed with saturated NH4C1 solution, brine, dried over anhydrous Na2S04 and concentrated under reduced pressure to get the residue. The crude product was purified via silica gel column chromatography with EtOAc and n-Hexane (1 :39) as an eluent to afford the desired compound (3.9 g, yield: 57%) as a white solid. 1H NMR (300 MHz, DMSO-d6): δ 7.76 (d, J = 8.4 Hz, 2H), 7.48 (d, J = 8.4 Hz, 2H), 7.18 (s, 1H), 4.14 (s, 2H), 2.40 (s, 3H), 2.01-1.90 (m, 4H), 1.77-1.68 (m, 2H), 1.30 (s, 9H).
Step 4: Synthesis of tert-butyl (1 -(pyrrolidin-1 -ylmethyl)cyclobutyl)carbamate: BocHN
To a stirred solution of (l-((tert-butoxycarbonyl)amino)cyclobutyl)methyl 4- methylbenzenesulfonate (step 3, 3.9 g, 10.1 mmol) in DMF (40 ml) under N2 atmosphere, were added K2C03 (4.5 g, 33 mmol), followed by pyrrolidine (1.8 ml, 22 mmol) at room temperature. The reaction mixture was heated to 110 °C and continued for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to room temperature, filtered through celite, was washed with EtOAc (twice). The filtrate was washed with water, brine, dried over anhydrous Na2S04 and concentrated under reduced pressure to give the residue. The crude product was purified via silica gel column chromatography with EtOAc and n-Hexane (50: 50) to afford the desired compound (2.3 g, yield: 8 %) as a thick oil. 1H MR (300 MHz, DMSO-d6): δ 6.63 (s, 1H), 3.22-3.17 (m, 2H), 2.75-2.69 (m, 2H), 2.39 (s, 2H), 2.22-2.11 (m, 2H), 1.96-1.88 (m, 2H), 1.80-1.76 (m, 2H), 1.65-1.59 (m, 4H), 1.37 (s, 9H).
Step 5: Synthesis of 1 -(pyrrolidin- 1 -ylmethyljcyclobutan- 1 -amine hydro chloride :
To a stirred solution of tert-butyl (1 -(pyrrolidin- l-ylmethyl)cyclobutyl)carbamate (step 4, 2.7 g, 10.6 mmol) in dioxane (10 ml), was added 6N HCl in dioxane (30 ml) and stirred for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to afford the desired product (2 g, yield: 100%). Next reaction was carried out without any further purification.
The below intermediates 53-56 were prepared by the procedure similar (including reagents and reaction conditions) to the above described in the synthesis of intermediate-52 by using suitable reagents, reactents and solvents at appropriate conditions.
Figure imgf000083_0001
Intermediate 57: Preparation of (S)-l-prolylpyrroridine: H O
Synthesis of tert-butyl (S)-2-(pyrrolidine-l -carbonyl)pyrrolidine-l -carboxyh
Figure imgf000084_0001
A solution of (tert-butoxycarbonyl)-L-proline (Intermediate 13 -step 1, 5.g, 23.0 mmol,
1.0 eq) in DMF (25 mL) was treated sequentially with HOBt (6.0 g, 35.0 mmol, 1.5 eq), EDCI (13.0 g, 69.0 mmol, 3.0 eq) and pyrrolidine (35.0 g, 46.0 mmol, 2.0 eq). The suspension was stirred for about 15 minutes. Then DIPEA (20 mL, 35.0 mmol, 1.5 eq) was added and the reaction mixture was stirred for about 12 hours. The reaction was quenched by pouring into 50 mL of ethyl acetate and water, extracting with ethylacetate, washed with water, saturated aqueous NaHC03, and a portion of brine. The solution was then dried over Na2S04, filter, and concentrated. The residue was purified by column chromatography on silica gel 2% MeOH and DCM to afford the desired compound (3.5 g, yield: 59.0%) as an off white solid. 1H NMR (DMSO-d6, 300 MHz): δ 4.38-4.29 (m, 1H), 3.51-3.23 (m, 6H), 2.19- 1.90 (m, 1H), 1.88-1.72 (m, 7H) and 0.69 (s, 9H); Mass: [M+Na]+ 291.16 (60%).
Step 2: Synthesis of (S)-l-prolylpyrrolidine:
To the stirred solution of tert-butyl (S)-2-(pyrrolidine-l-carbonyl)pyrrolidine-l- carboxylate (step 1, 3.5 g, 13.2 mmol, 1.0 eq) in 1,4-dioxane.HCl (30 mL) which was stirred at room temperature for about 4 hours. After completion of the reaction (monitored by TLC), the volatile solvent was concentrated to afford the desired compound as a brown liquid, which was without purification go to next step.
EXAMPLES
Example 1: Preparation of 2,2-dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR, 13aRJ3bR)-5a,5b,8,8 J la-pentamethyl-3a-((2-methyl-l-morpholinopropan-2-yl)carbamoyl)- l-(l-methylcvclopropyl)icosahvdro-lH-cvclo entaralchrvsen-9-yl)oxy)butanoic acid:
Figure imgf000084_0002
Step 1: Synthesis of (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla-penta methyl-3a-((2-methyl-l -morpholinopropan-2-yl)carbamoyl)-l -(1 -methylcyclopropyl) icosahydro-lH-cyclopenta[a ]chrysen-9-yl acetate:
Figure imgf000085_0001
To a stirred solution of 2-methyl-l-morpholinopropan-2-amine (Intermediate 1, 0.80 g, 4.75 mmol, 2.1 eq) and Triethylamine (10.0 mL, 18.79 mmol, 5.0 eq) in CH2C12 (20 ml) at 0°C was added (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-(chlorocarbonyl)-5a,5b, 8,8,l la-pentamethyl-l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (prepared as described in WO 2013/160810 A2, 1.2 g, 2.26 mmol, 1.0 eq) in CH2C12 (10 ml). The reaction mixture was allowed to stir at room temperature for overnight. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with water and extracted with CH2C12. The combined organic extracts were dried over Na2S04, filtered and evaporated under reduced pressure. The residue was purified by column chromatography by using 8% methanol: DCM as an eluent to afford the desired product (1.4 g, yield: 95.0%) as a semi solid. 1H NMR (DMSO-d6, 300 MHz): δ 4.40-4.35 (q, 1H), 3.55 (t, 4H), 3.45 (bs, 4H), 2.76 (s, 1H), 2.57 (m, 1H), 2.39-2.10 (m, 13H), 1.96 (m, 3H), 1.91 (m, 2H), 1.69-1.05 (s, 16H), 0.95 (s, 3H), 0.87 (s, 3H), 0.84 (s, 3H), 0.82 (m, 12H), 0.34-0.29 (t, 2H) and 0.19- 0.18 (q, 2H); Mass: [M]+ 694.39 (100%).
Step 2: Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-9-hydroxy-5a,5b,8,8, lla^entamethyl-N-(2-methyl-l-morpholinopropan-2-yl)-l-(l-methylcyclopropyl)icosahydro -3aH-cyclopenta[a]chrysene-3 -carboxamide:
Figure imgf000085_0002
To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8, 11 a-pentamethyl-3 a-((2-methyl- 1 -morpholinopropan-2-yl)carbamoyl) - 1 -( 1 -methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (step 1, 1.4 g, 2.15 mmol, 1.0 eq) in THF (7 ml) and Methanol (7 ml) was added potassium carbonate (2.0 g, 15.05 mmol, 7.0 eq). The reaction mixture was stirred at room temperature for about 48 hours. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was filtered through a pad of celite and washed with CH2C12. The filtrate was evaporated under reduced pressure and the crude was purified by column chromatography by using 1% methanol: dichloromethane as an eluent to afford the desired product (0.750 g, yield: 57.0%) as a white solid. 1H NMR (CDCI3, 300 MHz): δ 4.28-4.26 (d, 1H), 3.55 (t, 4H), 3.44 (bs, 4H), 3.17 (d, 1H), 2.98 (q, 1H), 2.78-2.63 (m, 2H), 2.35-2.31 (m, 12H), 2.05 (bd, 1H), 1.95-1.87 (m, 3H), 1.61-1.25 (m, 19H), 0.93 (s, 3H), 0.86 (s, 9H), 0.77 (s, 3H), 0.65 (m, 3H), 0.34-0.29 (t, 2H) and 0.19-0.14 (q, 2H); Mass: [M]+ 652.51 (100%).
Step 3: Synthesis of 2,2-dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR) -5a,5b,8,8,lla-pentamethyl-3a-((2-methyl-l-morpholinopropan-2-yl)carbamoyl)-l-(l -methyl cyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)butanoic acid:
To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy- 5a,5b,8,8, 1 la-pentamethyl-N-(2-methyl- l-morpholinopropan-2-yl)- l-(l-methylcyclopropyl) icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide (step 2, 0.30 g, 0.49 mmol, 1.0 eq) and 2,2-dimethyl succinicanhydride (0.251 g, 1.96 mmol, 4.0 eq) in toluene (10 ml) was added DMAP (0.120 g, 0.93 mmol, 2.0 eq). The reaction mixture was heated at 90°C for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was concentrated under reduced pressure, cooled to 0°C, acidified to pH~5 with IN HC1 and extracted with CH2CI2. The combined organic extracts were washed with water, dried over Na2S04, filtered and evaporated under reduced pressure. The crude residue was purified by column chromatography by using 5% methanol: dichloromethane as an eluent to afford the desired product (0.100 g, yield: 27.0%) as a white solid. 1H NMR (CDC13, 300 MHz): δ 4.40-4.35 (q, 1H), 3.57-3.54 (m, 4H), 3.45 (t, 4H), 2.42-2.3 5 (m, 14H), 2.05 (bs, 1H), 1.87 (m, 2H), 1.65-1.23 (m, 18H), 1.16 (m, 8H), 1.10- 0.94 (m, 2H), 0.87 (s, 3H), 0.84-0.78 (m, 16H), 0.32 (t,lH) and 0.19 (t, 2H); Mass: [M]~ 780.49 (100%).
The below examples 2-7 were prepared by the procedure similar to example- 1 by using corresponding intermediates inpresence of suitable reagents, reactents and solvents at appropriate reaction conditions. The corresponding intermediates and the charctrization data are mentioned in the below table.
Figure imgf000086_0001
1H NMR (CDC13, 300 MHz): δ
4.40-4.35 (q, IH), 3.57-3.54 (m,
4f 4H), 3.45 (t, 4H), 2.42-2.35 (m,
14H), 2.05 (bs, IH), 1.87 (m, 2H),
2 1.65-1.23 (m, 18H), 1.16 (m, 8H), o 1 1.10-0.94 (m, 2H), 0.87 (s, 3H),
0.84-0.78 (m, 16H), 0.32 (t,lH) and 0.19 (t, 2H); Mass: [M]~ 780.49 (100%).
1H NMR (CDCI3, 300 MHz): δ 4.40-4.35 (q, IH), 3.57-3.54 (m, 4H), 3.45 (t, 4H), 2.42-2.35 (m, 14H), 2.05 (bs, IH), 1.87 (m, 2H),
3 1.65-1.23 (m, 18H), 1.16 (m, 8H), o * 1.10-0.94 (m, 2H), 0.87 (s, 3H),
0.84-0.78 (m, 16H), 0.32 (t,lH) and 0.19 (t, 2H); Mass: [M]~ 780.49 (100%).
1H NMR (DMSO-d6, 300 MHz): δ 12.21 (s, IH), 6.68 (s, IH), 4.40 (d, 2H), 4.18 (q, IH), 3.24 (s, IH), 3.15 (t, IH), 2.42-2.31 (m, 6H), 2.08 (d, 3H), 1.86 (d, 3H), 1.75 (t,
4 7H), 1.54-1.34 (m, 13H), 1.25 (t,
12H), 1.16 (d, 12H), 1.09 (t, 3H), 0.97 (s, 3H), 0.95 (s, 6H), 0.83 (t, 9H), 0.33 (t, 2H) and 0.19 (t, 3H); Mass: [M]+ 866.72 (100%); HPLC: 88.63%. 1H NMR (DMSO-d6, 300 MHz): δ
12.13 (s, 1H), 4.40 (q, 1H), 4.18 / (bs, 1H), 3.50 (bs, 2H), 3.03 (bs,
3H), 2.43-2.22 (m, 4H), 1.87 (m, 8H), 1.59-1.34 (m, 7H), 1.24-1.15
5 39 HOOC (m, 10H), 1.10-1.06 (m, 12H),
0.96 (s, 3H), 0.93-0.78 (m, 16H), 0.33 (t, 2H) and 0.19 (t, 2H); Mass: [M]+ 725.69 (100%); HPLC: 87.05%.
1H NMR (DMSO-d6, 300 MHz): δ 12.17 (s, 1H), 4.46 (m, 1H), 4.40 (m, 1H), 3.56 (s, 4H), 3.45 (m, 2H), 3.23 (m, 2H), 2.65 (m, 1H), 2.30 (m, 1H), 2.27-1.59 (m, 9H),
6 57
1.54-1.23 (m, 13H), 1.16 (m,
Figure imgf000088_0001
15H), 0.94-0.71 (m, 18H), 0.31 (m, 2H) and 0.20-0.12 (m, 2H); Mass: [M]+ 749.66 (60%); HPLC: 87.22%.
1H NMR (DMSO-d6, 300 MHz): δ 12.13 (s, 1H), 4.39 (m, 1H), 4.14 (m, 1H), 3.50 (m, 2H), 2.78 (m, 3H), 2.40 (m, 3H), 2.24-2.08 (m, 4H), 1.91 (m, 3H), 1.73-1.63 (m,
7 13
HOOC — °* /C fl— 5H), 1.55-1.43 (m, 6H), 1.38-1.23
(m, 10H), 1.16-1.05 (m, 14H), 1.01-0.95 (m, 4H), 0.88-0.78 (m, 18H), 0.34 (m, 2H) and 0.21 (m, 2H); Mass: [M+H]+ 778.66 (100%); HPLC: 91.00%.
Example 8: Preparation of 5-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2- ((4-ethylpiperazin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a.5b.8.8 J la-pen tamethyl-l-(l- methylcvclopropyl)icosahvdro-lH-cvclopentaralchrvsen-9-yl)oxy)-3,3-dimethyl-5-oxo pentanoic acid:
Figure imgf000089_0001
Step:l Synthesis of (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-3a-((S)-2-((4-ethyl piperazin-l -yl)methyl)pyrrolidine-l -carbonyl)-5a,5b,8,8,l 1 a-pentamethyl-1 -(1 -methylcyclo propyl)icosahydro-lH-cyclopenta[a ]chrysen-9-yl acetate:
Figure imgf000089_0002
To a stirred solution of (S)-l-ethyl-4-(pyrrolidin-2-ylmethyl)piperazine (Intermediate
13, 1.4 g, 7.07 mmol, 1.5 eq) and triethylamine (3.3 niL, 23.5 mmol, 5.0 eq) in CH2CI2 (15 ml) at 0 °C was added (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-(chlorocarbonyl) -5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9- yl acetate (prepared as described in WO 2013/160810 A2, 2.5 g, 4.7 mmol, 1.0 eq) in CH2CI2 (10 ml). The reaction mixture was allowed to stir at room temperature for overnight. The reaction mixture was diluted with water and extracted with CH2CI2. The combined organic extracts were dried over Na2S04, filtered and evaporated under reduced pressure. The residue was purified by column chromatography by using 5% methanol: DCM as an eluent gave the desired product (1.2 g, yield: 37.0%) as a light yellow solid. 1H NMR (DMSO-d6, 300 MHz): δ 4.40-4.35 (m, 1H), 4.14 (m, 1H), 3.57 (m, 1H), 2.81 (m, 1H), 2.31-2.27 (m, 10H), 2.08 (m, 2H), 1.99 (s, 3H), 1.92-1.88 (m, 4H), 1.67-1.43 (m, 12H), 1.34-1.23 (m, 11H), 1.08-0.79 (m, 23H), 0.36 (m, 2H) and 0.21 (m, 2H); Mass: [M-H]+ 692.66 (100%).
Step 2: Synthesis of ((S)-2-((4-ethylpiperazin-l-yl)methyl)pyrrolidin-l-yl)((lR,3aS,5aR,5bR, 7aR,9S,l 1 aR,l lbR,l 3aR,l 3bR)-9-hydroxy-5a,5b, 8,8,11 a-pentamethyl-1 -(1 -methylcyclo propyl)icosahydro-3aH-cyclopenta[a]chrysen-3a-yl)methanone:
Figure imgf000089_0003
To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-
((4-ethylpiperazin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b, 8,8,1 la-pentamethyl-l-(l- methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (step 1, 1.2 g, 1.74 mmol, 1.0 eq) in THF (12 mL), Methanol (12 mL) and water (12 mL) was added LiOH (0.73 g, 17.4 mmol, 10.0 eq). The reaction mixture was stirred at room temperature for about 12 hours. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was diluted with water and extracted with DCM. The organic layer was washed with water and dried with Na2S04. The organic layer was evaporated under reduced pressure and the crude was purified by column chromatography by using 8% methanol: dichloromethane as an eluent gave the desired product (0.8 g, yield: 71.0%) as a white solid. 1H NMR (CDC13, 300 MHz): δ 4.26 (d, J=5.1 Hz, 1H), 4.14 (m, 1H), 3.49 (m, 1H), 3.01-2.94 (m, 1H), 2.80 (m, 1H), 2.32-2.30 (m, 9H), 2.03-1.70 (m, 9H), 1.71-1.50 (m, 8H), 1.46-1.19 (m, 12H), 1.15-0.72 (m, 21H), 0.65 (s, 3H), 0.34 (m, 2H) and 0.21 (m, 2H); Mass: [M-H]+ 650.62 (100%).
Step 3: Synthesis of 5-(((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-3a-((S)-2-((4-ethyl piperazin-l -yl)methyl)pyrrolidine-l -carbonyl)-5a,5b,8,8,l 1 a-pentamethyl-1 -(1 -methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)-3,3-dimethyl-5-oxopentanoic acid:
To a stirred solution of ((S)-2-((4-ethylpiperazin-l-yl)methyl)pyrrolidin-l-yl)((lR, 3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy-5a,5b,8,8,l la-pentamethyl-l-(l- methylcyclopropyl)icosahydro-3aH-cyclopenta[a]chrysen-3a-yl)methanone (step 2, 0.400 g, 0.6 mmol, 1.0 eq) and 4,4-dimethyldihydro-2H-pyran-2,6(3H)-dione (0.170 g, 1.2 mmol, 2.0 eq) in toluene (20 ml) was added DMAP (0.120 g, 4.8 mmol, 5.0 eq). The reaction mixture was heated at 90 °C for overnight. TLC indicated starting material was consumed and the desired product was observed. The mixture was concentrated under reduced pressure, cooled to Oo C, acidified to pH~5 with IN HC1 and extracted with CH2C12. The combined organic extracts were washed with water, dried over Na2S04, filtered and evaporated under reduced pressure. The crude residue was purified by column chromatography by using 10% methanol and dichloromethane as an eluent gave the desired product (0.130 g, yield: 35.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 11.97 (s, 1H), 4.41 (q, 1H), 4.13 (bs, 1H), 3.49-3.39 (m, 4H), 2.79 (m, 1H), 2.39-2.07 (m, 10H), 1.90-1.63 (m, 7H), 1.55-1.14 (m, 18H), 1.09-0.95 (m, 20H), 0.88-0.80 (m, 15H), 0.33 (m, 2H) and 0.19 (m, 2H). Mass: [M+H] 792.63 (100%). HPLC: 91.29%.
The below example 9 were prepared by the procedure similar to example-8 by using corresponding intermediates inpresence of suitable reagents, reactents and solvents at appropriate reaction conditions. The corresponding intermediates and the charctrization data are mentioned in the below table.
Figure imgf000091_0001
Example 10: Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((l-((R)-3-hvdroxypyrrolidin-l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b,8,8 J la-penta methyl- l-(l-methylcvclopropyl)icosahvdro-lH-cvclopentaralchrvsen-9-yl)oxy)carbonyl)-
Figure imgf000091_0002
Step 1: Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-3a-((l-((R)-3-
(benzyloxy)pyrrolidin-l-yl)-2-methylpropan-2-yl)carbamoyl)-5a
( 1 -methylcyclopropyl)icosahydro-lH-cyclopenta[a ]chrysen-9-yl acetate:
Figure imgf000091_0003
To a stirred solution of (R)-l-(3-(benzyloxy)pyrrolidin-l-yl)-2-methylpropan-2-amine
(Intermediate 11, 0.96 g, 3.9 mmol, 2.0 eq) and triethylamine (1.3 mL, 9.75 mmol, 5.0 eq) in
CH2C12 (40 ml) at 0 °C was added (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a- (chlorocarbonyl)-5a,5b,8,8, 1 la-pentamethyl- l-(l-methylcyclopropyl)icosahydro- lH-cyclo penta[a]chrysen-9-yl acetate (prepared as described in WO 2013/160810 A2, 1.0 g, 1.90 mmol, 1.0 eq) in CH2CI2 (24 ml). The reaction mixture was allowed to stir at room temperature for overnight. The reaction mixture was diluted with water and extracted with CH2CI2. The combined organic extracts were dried over Na2S04, filtered and evaporated under reduced pressure. The residue was purified by column chromatography by using 4% methanol: DCM as an eluent to afford the desired product (1.0 g, yield: 71.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 7.33-7.29 (m, 5H), 6.71 (s, 1H), 4.41 (s, 2H), 4.04 (s, 1H), 2.96 (t, 2H), 2.78-2.65 (m, 3H), 2.10-2.08 (m, 9H), 1.99 (s, 3H), 1.44-1.13 (m, 19H), 1.08-1.94 (m, 5H), 0.91 (s, 3H), 0.86 (s, 10H), 0.77 (m, 3H), 0.64 (s, 4H), 0.30 (d, 2H) and 0.18 (d, 2H); Mass: [M]+744.62 (100%).
Step 2: Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-N-(l-((R)-3- (benzyloxy)pyrrolidin-l-yl)-2-methylpropan-2-yl)-9-hydroxy-5a,5b,8,8,lla-pentamethyl-l-(l- methylcyclopropyl)icosahyd -3aH-cyclopenta[a]chrysene-3a-carboxamide:
Figure imgf000092_0001
To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-((R)- 3-(benzyloxy)pyrrolidin- l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b,8,8, 1 la-pentamethyl- l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (step 1, 1.0 g, 1.34 mmol, 1.0 eq) in THF (10 ml) and Methanol (10 ml) was added potassium carbonate (1.3 g, 9.43 mmol, 7.0 eq). The reaction mixture was stirred at room temperature for about 48 hours. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was filtered through a pad of celite and washed with CH2CI2. The filtrate was evaporated under reduced pressure and the crude was purified by column chromatography by using 4% methanol: dichloromethane as an eluent to afford the desired product (0.800 g, yield: 84.0%) as a white solid. 1H NMR (CDCI3, 300 MHz): δ 7.35-7.26 (m, 5H), 6.70 (s, 1H), 4.45 (s, 2H), 4.25 (d, 1H), 4.04 (s, 1H), 2.96 (t, 2H), 2.81-2.65 (m, 3H), 2.10-1.55 (m, 9H), 1.44-1.13 (m, 19H), 1.08-1.94 (m, 5H), 0.91(s, 3H), 0.86 (s, 10H), 0.77 (m, 3H), 0.64 (s, 4H), 0.30 (d, 2H) and 0.18 (d, 2H); Mass: [M]+ 701.56 (100%).
Step 3: Synthesis of 1-benzyl 3-((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-3a-((l- ((R)-3-(benzyloxy)pyrrolidin-l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b,8,8,lla- pentamethyl-1 -( 1 -methylcyclopropyl)icosahydro-l H-cyclopenta[a] chrysen-9-yl) (lR,3S)-2,2- dimethylcyclobutane- -dicarboxylate:
Figure imgf000093_0001
To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-N-(l-((R)- 3-(benzyloxy)pyrrolidin-l-yl)-2-methylpropan-2-yl)-9-hydroxy-5a,5b,8,8,l la-pentamethyl-l- (l-methylcyclopropyl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide (step 2, 0.800 g, 2.14 mmol, 1.0 eq) in DCM (40 ml) then added (lS,3R)-3-((benzyloxy)carbonyl)-2,2- dimethylcyclobutane-l-carboxylic acid (prepared as described in WO 2011/007230 A2, 0.45 g, 1.71 mmol, 1.5 eq) and the reaction mixture was cooled to 0 °C, then added DCC (0.47 g, 2.28 mmol, 2.0 eq) followed by DMAP (0.020 g, 0.22 mmol, 0.2 eq). The reaction mixture was raised to room temperature and stirred for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was evaporated under reduced pressure and diluted with water. The aqueous layer was extracted with CH2CI2. The organic layer was washed with water and brine solution. The combined organic layers were dried over Na2S04, filtered and evaporated under reduced pressure. The crude residue was purified by column chromatography by using 3% MeOH and DCM as an eluent to obtain the desired product (0.400 g, yield: 40.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 7.36 (t, 10H), 6.71 (s, 1H), 5.14 (q, 4H), 4.41 (t, 1H), 4.04 (t, 1H), 2.79-2.65 (m, 8H), 2.15- 2.06 (m, 2H), 1.94-1.73 (m, 4H), 1.70-1.46 (m, 7H), 1.43-1.34 (m, 10H), 1.26-1.21 (m, 12H), 1.11 (s, 1H), 1.03-1.01 (m, 2H), 0.94 (s, 3H), 0.91 (s, 3H), 0.87 (s, 6H), 0.82 (s, 9H), 0.32 (d, 2H) and 0.19 (d, 2H); Mass: [M]+ 846.58 (100%).
Step 4: Synthesis of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-3a-((l- ((R)-3-hydroxypyrrolidin-l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b,8,8,lla-pentamethyl- 1-(1 -methylcyclopropyl)icosahydro-lH-cyclopenta[a ] chrysen-9-yl)oxy )carbonyl)-2,2- dimethylcyclobutane-l-carboxylic acid:
To a solution of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a- ((l-((R)-3-(benzyloxy)pyrrolidin-l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b, 8,8,1 la- pentamethyl-l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2- dimethylcyclobutane-l,3-dicarboxylate (step 3, 0.400 g, 0.42 mmol, 1.0 eq) in MeOH (10 mL) and ethylacetate (20 mL) was added palladium carbon (0.026 g, 0.25 mmol, 0.5 eq). The reaction mixture was stirred in hydrogen atmosphere at room temperature for overnight. The reaction mixture was filtered through a pad of celite and was washed with MeOH and DCM.
The filtrate was evaporated under reduced pressure and the crude residue was purified by column chromatography by using 5% methanol: DCM as an eluent to afford the desired compound (0.100 g, yield: 27.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 12.15 (s, 1H), 6.71 (s, 1H), 4.69 (d, 1H), 4.37 (t, 1H), 4.14 (s, 1H), 2.79-2.65 (m, 8H), 2.15-2.06 (m, 2H), 1.94-1.73 (m, 4H), 1.70-1.46 (m, 7H), 1.43-1.34 (m, 10H), 1.26-1.21 (m, 12H), 1.11 (s, 1H), 1.03-1.01 (m, 2H), 0.94 (s, 3H), 0.91 (s, 3H), 0.87 (s, 6H), 0.82 (s, 9H), 0.32 (d, 2H) and 0.19 (d, 2H); Mass: [M]+ 765.50 (100%); HPLC: 89.97%.
The below examples 11-54 were prepared by the procedure similar to example- 10 by using corresponding intermediates inpresence of suitable reagents, reactents and solvents at appropriate reaction conditions. The corresponding intermediates and the charctrization data are mentioned in the below table.
Figure imgf000094_0001
Figure imgf000095_0001
Figure imgf000096_0001
Figure imgf000097_0001
1H NMR (DMSO-d6, 300 MHz): δ
12.15 (bs, IH), 4.35 (m, 2H), 3.59 (m, IH), 3.48 (m, IH), 3.18 (m, IH), 3.09 (m, 2H), 2.82-2.73 (m, 3H), 2.35-2.24 (m, 2H), 1.95-1.80
17 (m, 11H), 1.59-1.47 (m, 11H),
1.40-1.26 (m, 10H), 1.24-1.03 (m, 10H), 0.96-0.81 (m, 21H), 0.35 (m, 2H) and 0.23 (m, 2H); Mass: [M+H]+ 789.77 (100%); HPLC: 96.66%.
1H NMR (DMSO-d6, 300 MHz): δ 12.11 (bs, IH), 7.53 (s, IH), 7.06 (s, IH), 6.89 (s,lH), 4.35 (t, IH), 4.20 (m, IH), 4.06 (m, 2H), 3.50 (m, IH), 2.82 (q, 2H), 2.64 (m,
47 IH), 2.37-2.20 (m, 2H), 1.92-1.86
HOOCT (m, 4H), 1.62-1.33 (m, 16H), 1.26- 1.23 (m, 6H), 1.09-1.06 (m, 2H), 0.96-0.82 (m, 25H), 0.36 (m, 2H) and 0.23 (m, 2H); Mass: [M+H] 758.65 (100%); HPLC: 91.00%
Figure imgf000099_0001
Figure imgf000100_0001
1H NMR (DMSO-d6, 300 MHz): δ
12.05 (s, 1H), 4.37 (t, J=7.8 Hz, 1H), 4.13 (m, 1H), 3.50 (m, 2H), 2.81-2.73 (m, 4H), 2.33-2.20 (m, 5H), 2.05 (m, 1H), 2.33-2.20 (m, 5H), 2.05 (m, 1H), 1.91-1.84 (m,
19
HOOCT 4H), 1.72-1.62 (m, 5H), 1.72-1.44
(m, 9H), 1.39-1.24 (m, 12H), 1.11- 0.08 (m, 3H), 0.95 -0.81 (m, 27H), 0.35 (m, 2H) and 0.21 (m, 2H); Mass: [M-H]+ 832.61 (100%); HPLC: 94.40%.
1H NMR (DMSO-d6, 300 MHz): δ 12.13 (s, 1H), 4.37 (t, J=7.8 Hz, 1H), 4.14 (m, 1H), 3.49-3.40 (m, 1H), 2.8-2.70 (m, 4H), 2.28-2.20 (m, 3H), 1.99-1.80 (m, 7H), 1.71-
20 1.63 (m, 6H), 1.56-1.45 (m, 8H),
HOOCT
1.35-1.24 (m, 15H), 1.10-1.04 (m, 5H), 0.95-0.81 (m, 24H), 0.34 (m, 2H) and 0.20 (m, 2H); Mass: [M+H]+ 789.64 (100%); HPLC: 90.19%.
1H NMR (DMSO-d6, 300 MHz): δ 12.05 (s, 1H), 4.37 (t, J=7.8 Hz, 1H), 4.13 (m, 1H), 3.50 (m, 2H), 2.81-2.73 (m, 3H), 2.60-2.57 (m, 1H), 2.41 (m, 4H), 2.33-2.20 (m,
21 5H), 2.14-2.00 (m, 2H), 1.91-1.84
Figure imgf000101_0001
(m, 4H), 1.72-1.44 (m, 11H), 1.34- 1.23 (m, 11H), 1.11-1.05 (m, 3H), 0.95-0.81 (m, 30H), 0.34 (m, 2H) and 0.19 (m, 2H); Mass: [M+H]+ 818.64 (100%); HPLC: 94.00%. 1H MR (300 MHz, DMSO-d6): δ
12.16 (brs, IH), 11.89 (brs, IH), 7.74-7.13 (m, 6H), 4.37-4.32 (m, IH), 4.11 (m, IH), 3.47 (m, 3H),
27 2.82-2.76 (m, 3H), 2.35-1.12 (m,
40H), 0.93-0.81 (m, 22H), 0.34-
Figure imgf000102_0001
0.29 (m, 2H), 0.21-0.17 (m, 2H); ES Mass: 903.65 [M+H]+; HPLC: 77% +13% (isomer).
1H NMR (DMSO-d6, 300 MHz): δ 12.14 (s, IH), 8.23-8.24 (m, IH), 7.94-7.96 (m, IH), 7.37 (bs, IH), 7.06-7.09 (m, IH), 6.48-6.49 (s, IH), 4.22-4.43 (m, 4H), 3.50 (bs, IH), 3.37 (bs, IH), 2.73-2.82 (m, 3H), 2.25-2.31 (m, 2H), 2.07 (s,
51
HOOCT IH), 1.86-1.97 (m, 3H), 1.57-1.66
(m, 13H), 1.23-1.45 (m, 13H), 1.05-1.14 (m, 3H), 0.96-0.99 (bs, 4H), 0.81-0.91 (m, 20H), 0.32- 0.36 (t, 2H) and 0.18-0.21 (m, 2H); Mass: [M]+ 807.41 (100%); HPLC: 95.34%.
1H NMR (DMSO-d6, 300 MHz): δ 12.00 (bs, IH), 4.38 (t, IH), 4.17 (m, IH), 3.50 (m, 2H), 3.09 (q, 4H), 2.82 (q, 4H), 2.31 (m, IH),
22 2.06-1.83 (m, 8H), 1.66-1.34 (m,
HOOCT 15H), 1.26-1.06 (m, 14H), 0.95- 0.81 (m, 21H), 0.33 (m, 2H) and 0.21 (m, 2H); Mass: [M+H] 775.64 (100%); HPLC: 92.00%.
Figure imgf000103_0001
Figure imgf000104_0001
1H MR (300 MHz, DMSO-d6): δ
11.86 (brs, 1H), 4.37-4.32 (m,
- > n 1H), 4.10-4.01 (m, 3H), 3.71-3.49
(m, 3H), 3.41-3.37 (m, 2H), 2.95-
30 1.07 (m, 47H), 0.95-0.81 (m,
HO
21H), 0.32 (m, 2H), 0.19 (m, 2H); ES Mass: 847.57 [M+H]+; HPLC: 94.47%.
1H NMR (300 MHz, DMSO-d6): δ 12.16 (brs, 1H), 4.38-4.32 (m, 1H), 4.17-4.14 (m, 1H), 3.82-3.78 (m, 1H), 3.56-3.53 (m, 1H), 3.36-
40 2.03 (m, 26H), 2.09-1.13 (m,
24H), 1.08-0.85 (m, 24H), 0.45- 0.43 (m, 1H), 0.35-0.33 (m, 1H), 0.22-0.20 (m, 2H); ES Mass: 830.68 [M+H]+.
1H NMR (DMSO-d6, 300 MHz): δ 12.14 (s, 1H), 4.37 (t, J=7.8 Hz, 1H), 4.09 (m, 1H), 3.48-3.46 (m, 2H), 3.09-2.90 (m, 4H), 2.81-2.75 (m, 3H), 2.35-2.25 (m, 4H), 2.11-
2.04 (m, 3H), 1.91-1.73 (m, 9H),
23 1.65-1.53 (m, 7H), 1.44-1.30 (m,
Figure imgf000105_0001
9H), 1.26-1.19 (m, 10H), 1.11-
1.05 (m, 4H), 1.11-1.05 (m, 4H), 0.95 (s, 3H), 0.91-0.81 (m, 18H), 0.34 (m, 2H) and 0.21-0.16 (m, 2H); Mass: [M+H]+ 858.68 (100%); HPLC: 96.71%. 1H NMR (DMSO-d6, 300 MHz): δ
12.06 (s, 1H), 4.37 (t, J=7.8 Hz, 1H), 4.13 (m, 1H), 3.42-3.40 (m, 5H), 3.21 (s, 3H), 2.79-2.73 (m,
24 2H), 2.44-2.31 (m, 9H), 1.92-1.46
HOOCT
(m, 17H), 1.34-1.08 (m, 16H), 0.94 -0.81 (m, 24H), 0.34 (m, 2H) and 0.20 (m, 2H); Mass: [M+H]+ 834.67 (100%); HPLC: 98.18%.
1H NMR (DMSO-d6, 300 MHz): δ 12.08 (s, 1H), 4.35 (t, J=7.8 Hz, 1H), 4.10 (m, 1H), 3.50 (m, 2H), 2.82 (m, 2H), 2.34-1.87 (m, 5H),
25 1.73-1.56 (m, 16H), 1.43-1.03 (m,
HOOCT
22), 0.93-0.82 (m, 25H), 0.37 (m, 2H) and 0.21 (m, 2H); Mass: [M+H]+ 790.64 (100%); HPLC: 96.43%.
1H NMR (DMSO-d6, 300 MHz): δ 12.04 (s, 1H), 4.37 (t, J=7.8 Hz, 1H), 4.11 (m, 1H), 3.47 (m, 1H), 2.79-2.72 (m, 6H), 2.55-2.47 (m, 7H), 2.34-2.17 (m, 2H), 1.89-1.83
26
Hoocr (m, 4H), 1.72-1.53 (m, 9H), 1.45- 1.19 (m, 13H), 1.13-1.06 (m, 4H), 0.95-0.81 (m, 23H), 0.35 (m, 2H) and 0.20 (m, 2H); Mass: [M+H]+ 793.56 (100%); HPLC: 97.82%.
Figure imgf000107_0001
Figure imgf000108_0001
Example 55: Preparation of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 3a-((S)-2-((l J-dioxidothiomorpholino)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8 J la-penta methyl- l-(l-methylcyclopropyl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)carbonyl)-
Figure imgf000108_0002
Step:l Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla-penta methyl-1 -( 1 -methylcyclopropyl)-3a-( (S)-2-( thiomorpholinomethyl)pyrrolidine-l -carbonyl) icosahydro-lH-cyclopenta[a ]chrysen-9-yl acetate
Figure imgf000109_0001
To a stirred solution of (S)-4-(pyrrolidin-2-ylmethyl)thiomorpholine (Intermediate 26, 2.5 g, 13.48 mmol, 1.3 eq) and triethylamine (7.2 mL, 51.85 mmol, 5.0 eq) in CH2C12 (20 ml) at O °C was added (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-(chlorocarbonyl)-5a, 5b,8,8,l la-pentamethyl-l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (prepared as described in WO 2013/160810 A2, 5.5 g, 10.37 mmol, 1.0 eq) in CH2C12 (40 ml). The reaction mixture was allowed to stir at room temperature for overnight. The reaction mixture was diluted with water and extracted with CH2C12. The combined organic extracts were dried over Na2S04, filtered and evaporated under reduced pressure. The residue was purified by column chromatography by using 15% EtOAc and hexane as an eluent gave the desired product (6.0 g, yield: 85.0%) as an off white solid. 1H NMR (DMSO-d6, 300 MHz): δ 4.40 (m, 1H), 4.10 (m, 1H), 3.47-3.42 (m, 1H), 2.75 (m, 3H), 2.55 (m, 6H), 2.40- 2.20 (m, 3H), 1.99 (s, 3H), 1.93-1.44 (m, 12H), 1.39-1.17 (m, 11H), 1.10-0.95 (m, 6H), 0.88- 0.79 (m, 17H), 0.35-0.30 (m, 2H) and 0.21-0.16 (m, 2H); Mass: [M+H]+ 681.48 (100%). Step:2 Synthesis of ((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-9-hydroxy-5a,5b,8,8, lla-pentamethyl-l-(l-methylcyclopropyl)icosahydro-3aH-cyclopenta[a]chrysen-3a-yl)((S)- 2-( thiomorpholinomethyl)pyrrolidin-l -yl)methanone:
Figure imgf000109_0002
To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8, 11 a-pentamethyl- 1 -( 1 -methylcyclopropyl)-3a-((S)-2-(thiomorpholinomethyl)pyrrolidine- 1 - carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (step 1, 6.0 g, 8.8 mmol, 1.0 eq) in THF (60 mL), Methanol (60 mL) and water (20 mL) was added LiOH (3.7 g, 88.0 mmol, 10.0 eq). The reaction mixture was stirred at room temperature for about 12 hours. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was distilled out completely under vacuum to get crude residue. The residue was purified by column chromatography by using 15% EtOAc and hexane as an eluent gave the desired product (5.50 g, yield: 98.0%) as an off white solid. 1H NMR (CDC13, 300 MHz): δ 4.40 (m, 1H), 4.10 (m, 1H), 3.47-3.42 (m, 1H), 2.75 (m, 3H), 2.55 (m, 6H), 2.40-2.20 (m, 4H), 1.93-1.44 (m, 12H), 1.39-1.17 (m, 11H), 1.10-0.95 (m, 6H), 0.88-0.79 (m, 17H), 0.35- 0.30 (m, 2H) and 0.21-0.16 (m, 2H); Mass: [M+H]+ 639.51 (100%).
Step 3: Synthesis of 1 -benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8, lla-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-(thiomorpholinomethyl)pyrrolidine-l- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-dimethylcyclobutane-l,3- dicarboxylate:
Figure imgf000110_0001
To a stirred solution of ((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy -5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclopropyl)icosahydro-3aH-cyclopenta[a]chrysen- 3a-yl)((S)-2-(thiomorpholinomethyl)pyrrolidin-l-yl)methanone (step 2, 5.50 g, 8.62 mmol, 1.0 eq) in DCM (60 mL) then added (lR,3S)-3-((benzyloxy)carbonyl)-2,2-dimethylcyclo butane- 1-carboxylic acid (prepared as described in WO 2011/007230 A2, 3.38 g, 12.93 mmol, 1.5 eq), this reaction mixture was cooled to 0 °C then added DCC (3.55 g, 17.24 mmole, 2.0 eq) followed by DMAP (0.21 g, 1.72 mmol, 0.2 eq). The reaction mixture was raised to room temperature stirred for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was evaporated under reduced pressure and diluted with water. The aqueous layer was extracted with CH2CI2. The organic layer was washed with water and brine solution. The combined organic layers were dried over Na2S04, filtered and evaporated under reduced pressure. The crude residue was purified by column chromatography by using 15% EtOAc and Hexane as an eluent to obtain the desired product (5.50 g, yield: 72.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 7.37-7.34 (m 5H), 5.10 (q 2H), 4.33 (t, J=7.8 Hz, 1H), 4.08 (m, 1H), 3.46-3.37 (m, 3H), 3.03 (m, 4H), 2.93-2.72 (m, 7H), 2.34-2.17 (m, 2H), 1.89-1.83 (m, 4H), 1.72-1.53 (m, 9H), 1.45- 1.19 (m, 13H), 1.13-1.06 (m, 4H), 0.95-0.81 (m, 23H), 0.35 (m, 2H) and 0.20 (m, 2H); Mass: [M+H]+ 883.63 (20%).
Step 4: Synthesis of 1 -benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-3a-((S)-2- ((1,1 -dioxidothiomorpholino )methyl )pyrrolidine-l -carbonyl)-5a,5b, 8, 8, 11 a-pentamethyl-1 - (l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-dimethylcyclo butane -1,3 -dicarboxyla
Figure imgf000111_0001
solution 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,1 la-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-(thiomorpholinomethyl) pyrrolidine - 1 -carbonyl)icosahydro- 1 H-cyclopenta[a] chry sen-9-yl) ( 1 R,3 S )-2,2-dimethylcyclobutane- 1,3- dicarboxylate (step 3, 1.2 g, 1.36 mmol, 1.0 eq) was dissolved in dry CH2CI2 (20 ml), cooled to 0 °C, and treated with MCPBA (0.468 g, 2.72 mmol, 2.0 eq). The reaction was stirred at 0 °C for about 1.5 hours. The reaction mixture was diluted with EtOAc, washed with saturated aqueous NaHC03, brine, dried over MgS04, and filtered. The filtrate was evaporated under reduced pressure, the crude residue was purified by column chromatography by using 3% MeOH and DCM as an eluent to obtain the desired product (0.7 g, yield: 56.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 7.37-7.34 (m 5H), 5.10 (q 2H), 4.33 (t, J=7.8 Hz, 1H), 4.08 (m, 1H), 3.46-3.37 (m, 3H), 3.03 (m, 4H), 2.93-2.72 (m, 7H), 2.34-2.17 (m, 2H), 1.89-1.83 (m, 4H), 1.72-1.53 (m, 9H), 1.45-1.19 (m, 13H), 1.13-1.06 (m, 4H), 0.95-0.81 (m, 23H), 0.35 (m, 2H) and 0.20 (m, 2H); Mass: [M+H]+ 915.50 (100%).
Step 5: Synthesis of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-3a-((S)-2- ((1,1 -dioxidothiomorpholino )methyl )pyrrolidine-l -carbonyl)-5a,5b, 8, 8, 11 a-pentamethyl-1 - (l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl cyclo butane- 1-carboxylic acid:
To a solution of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a- ((S)-2-((l,l-dioxidothiomorpholino)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-penta methyl- l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-di methylcyclobutane-l,3-dicarboxylate (step 4, 0.700 g, 0.765 mmol, 1.0 eq) in ethylacetate (10 mL) was added Pd/C (0.040 g, 0.382 mmol, 0.5 eq). The reaction mixture was stirred in hydrogen atmosphere at room temperature for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was filtered through a pad of celite and was washed with MeOH and DCM. The filtrate was evaporated under reduced pressure, the crude residue was purified by column chromatography by using 5%
MeOH and DCM as an eluent to obtain the desired product (0.176 g, yield: 28.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 12.10 (s, 1H), 4.37 (t, J=7.8 Hz, 1H), 4.08 (m, 1H), 3.46-3.37 (m, 3H), 3.03 (m, 4H), 2.93-2.72 (m, 7H), 2.34-2.17 (m, 2H), 1.89-1.83 (m, 4H), 1.72-1.53 (m, 9H), 1.45-1.19 (m, 13H), 1.13-1.06 (m, 4H), 0.95-0.81 (m, 23H), 0.35 (m, 2H) and 0.20 (m, 2H); Mass: [M+H]+ 825.56 (100%); HPLC: 93.62%.
Example 56: Synthesis of (lS,3R)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR, 13aR,13bR)-5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-(pyrrolidin-l-yl methyl)pyrrolidine-l-carbonyl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)carbonyl) cyclobutane-l-carboxylic acid:
Figure imgf000112_0001
Step 1: Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla-penta methyl-1 -( 1 -methylcyclopropyl)-3a-( (S)-2-(pyrrolidin-l -ylmethyl)pyrrolidine-l -carbonyl) icosahydro-lH-cyclopenta[a ]chrysen-9-yl acetate:
Figure imgf000112_0002
To a stirred solution of (S)-l-(pyrrolidin-2-ylmethyl)pyrrolidine (Intermediate 39, 1.2 g, 7.8 mmol, 2.0 eq) and triethylamine (3.0 mL, 19.5 mmol, 5.0 eq) in CH2C12 (20 ml) at 0 °C was added (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-(chlorocarbonyl)-5a,5b,8,8, l la-pentamethyl-l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (prepared as described in WO 2013/160810 A2, 2.0 g, 3.9 mmol, 1.0 eq) in CH2C12 (20 ml). The reaction mixture was allowed to stir at room temperature for overnight. The reaction mixture was diluted with water and extracted with CH2C12. The combined organic extracts were dried over Na2S04, filtered and evaporated under reduced pressure. The residue was purified by column chromatography by using 4% methanol: DCM as an eluent gave the desired product (1.6 g, yield: 64.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 4.40 (m, 1H), 4.14 (m, 1H), 3.52 (m, 1H), 3.16 (m, 1H), 2.97 (m, 1H), 2.80 (m, 1H), 2.40 (m, 3H), 2.26 (m, 2H), 1.99 (s, 3H), 1.90-1.72 (m, 6H), 1.6 (m, 5H), 1.57-1.34 (m, 7H), 1.30-1.04 (m, 13H), 0.93 (m, 3H), 0.87 (m, 9H), 0.65 (s, 3H), 0.63 (s, 3H), 0.37 (m, 2H) and 0.22 (m, 2H); Mass: [M]+ 649.66 (100%). Step 2: Synthesis of ((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-9-hydroxy-5a,5b,8,8, lla-penta methyl-l-(l-methylcyclopropyl)icosahydro-3aH-cyclopenta[a]chrysen-3a-yl)((S)- 2-(pyrrolidin-l-ylmethyl)pyrrolidin-l- l methanone:
Figure imgf000113_0001
To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8, 1 la-pentamethyl- l-(l-methylcyclopropyl)-3a-((S)-2-(pyrrolidin- l-ylmethyl)pyrrolidine- 1- carbonyl) icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (step 1, 1.6 g, 2.5 mmol, 1.0 eq) in THF (16 ml) and Methanol (16 ml) and water (16 mL) was added Lithium hydroxide (1.2 g, 24.0 mmol, 10.0 eq). The reaction mixture was stirred at room temperature for about 12 hours. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was concentrated and this reaction mixture was diluted with water solid product was obtained which solid was filtered and dried. The obtained crude was purified by column chromatography by using 5% methanol and dichloromethane as an eluent gave the desired product (1.0 g, yield: 71.0%) as a white solid. 1H NMR (CDC13, 300 MHz): δ 4.28 (m, 1H), 4.10 (m, 1H), 3.49 (m, 1H), 3.16 (m, 1H), 2.97 (m, 1H), 2.80 (m, 1H), 2.40 (m, 3H), 2.26 (m, 2H), 1.90-1.72 (m, 7H), 1.6 (m, 5H), 1.57-1.34 (m, 7H), 1.30-1.04 (m, 13H), 0.93 (s, 3H), 0.87 (m, 9H), 0.65 (s, 3H), 0.63 (s, 3H), 0.37 (m, 2H) and 0.22 (m, 2H); Mass: [M]+ 607.55 (100%).
Step 3: Synthesis of 1 -benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8, lla-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidine-l- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)(lS,3R)-2,2-dimethylcyclobutane-l,3- dicarboxylate:
Figure imgf000113_0002
To a stirred solution of ((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy
-5a,5b, 8, 8,1 la-pentamethyl- l-(l-methylcyclopropyl)icosahydro-3aH-cyclopenta[a]chrysen-
3a-yl)((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidin-l-yl)methanone (step 2, 1.0 g, 1.6 mmol, 1.0 eq) in DCM (20 ml) then added (lR,3S)-3-((benzyloxy)carbonyl)-2,2-dimethylcyclobutane-
1-carboxylic acid (prepared as described in WO 2014/105926 Al, 0.600 g, 2.4 mmol, 1.5 eq) this reaction mixture was cooled to 0 °C then added DCC (0.670 g, 3.0 mmol, 2.0 eq) followed by DMAP (0.040 g, 0.3 mmol, 0.2 eq). The reaction mixture was raised to room temperature and stirred for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was evaporated under reduced pressure and diluted with water. The aqueous layer was extracted with CH2CI2. The organic layer was washed with water and brine solution. The combined organic layers were dried over Na2S04, filtered and evaporated under reduced pressure. The crude residue was purified by column chromatography by using 6% MeOH and DCM as an eluent to obtain the desired product (0.700 g, yield: 54.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 7.36 (m, 5H), 5.14 (m, 2H), 4.37 (t, 1H), 4.17 (m, 1H), 3.48 (m, 2H), 2.99 (m, 4H), 2.82 (m, 4H), 2.34 (m, 3H), 1.88 (m, 11H), 1.65-1.09 (m, 22H), 1.06-0.88 (m, 23H), 0.86 (m, 2H) and 0.33 (m, 2H); Mass: [M]+ 851.59 (100%).
Step 4: Synthesis of (lS,3R)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR, 13bR)-5a,5b,8,8,l 1 a-pentamethyl-1 -(1 -methylcyclopropyl)-3a-((S)-2-(pyrrolidin-l -ylmethyl) pyrrolidine-1 -carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane- 1 -carboxylic acid:
To a solution of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,l la-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidine- l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lS,3R)-2,2-dimethylcyclobutane-l,3- dicarboxylate (step 3, 0.700 g, 0.82 mmol, 1.0 eq) in ethylacetate (20 mL) was added Pd/C (0.055 g, 0.49 mmol, 0.6 eq). The reaction mixture was stirred in hydrogen atmosphere at room temperature for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was filtered through a pad of celite and was washed with MeOH and DCM. The filtrate was evaporated under reduced pressure, the crude residue was purified by column chromatography by using 8% methanol and DCM as an eluent gave the desired compound (0.210 g, yield: 34.0%) as an off white solid. 1H NMR (DMSO-d6, 300 MHz): δ 12.11 (s, 1H), 4.38 (t, 1H), 4.16 (m, 1H), 3.52 (m, 2H), 2.87-2.79 (m, 4H), 2.78 (q, 4H), 2.34 (q, 3H), 1.88 (bs, 11H), 1.65-1.09 (m, 23H), 1.06-0.88 (m, 23H), 0.33 (m, 2H) and 0.20 (m, 2H); Mass: [M]+ 761.71 (100%); HPLC: 95.03%.
The below examples 57-60 were prepared by the procedure similar to example-56 by using corresponding intermediates inpresence of suitable reagents, reactents and solvents at appropriate reaction conditions. The corresponding intermediates and the charctrization data are mentioned in the below table. Inter
Ex
media Structure Characterization data no.
te no
1H NMR (DMSO-d6, 300 MHz): δ 12.05 (s, 1H), 4.37 (t, J=7.8 Hz, 1H), 4.11 (m, 1H), 3.49 (m, 2H), 2.86 (m, 3H), 2.33-2.20 (m, 7H), 2.08 (m, 2H), 1.92-1.86 (m, 4H),
57 13 1.72-1.63 (m, 6H), 1.55-1.52 (m,
6H), 1.44-1.32 (m, 6H), 1.25-1.25 (m, 7H), 1.12-0.97 (m, 7H), 0.95- 0.80 (m, 24H), 0.34 (m, 2H) and 0.21 (m, 2H); Mass: [M+H]+ 804.60 (100%); HPLC: 91.00%.
1H NMR (300 MHz, CDC13): δ 4.75-4.58 (m, 1H), 4.47-4.44 (m, 1H), 4.33 (m, 1H), 3.52-3.49 (m,
58 41 2H), 3.1-1.25 (m, 45H), 1.05-0.78
(m, 22H), 0.49-0.46 (m, 1H), 0.34- 0.31 (m, 1H), 0.23-0.21 (m, 2H); ES Mass: 793.5 [M+H]+.
1H NMR (DMSO-d6, 300 MHz): δ 12.13 (s, 1H), 4.37 (t, J=7.8 Hz, 1H), 4.17 (m, 1H), 3.54-3.41 (m, 6H), 2.81-2.72 (m, 3H), 2.45-2.42 (m, 2H), 2.33-2.20 (m, 5H), 2.08 (m, 1H), 1.91-1.84 (m, 4H), 1.73-
59 16
HOOCT J--,, 1.62 (m, 5H), 1.55-1.44 (m, 6H),
1.34-1.29 (m, 6H), 1.26-1.19 (m, 7H), 1.11-0.99 (m, 3H), 0.96-0.81 (m, 21H), 0.34 (m, 2H) and 0.20 (m, 2H); Mass: [M+H]+ 777.60 (100%); HPLC: 94.00%. H NMR (DMSO-d6, 300 MHz): δ
12.11 (s, 1H), 4.38 (t, 1H), 4.16
1H), 3.52 (m, 2H), 2.87-2.79
(m, 4H), 2.78 (q, 4H), 2.34 (q,
60 28 3H), 1.88 (bs, 11H), 1.65-1.09 (m,
Figure imgf000116_0001
23H), 1.06-0.88 (m, 23H), 0.32
(m, 2H) and 0.19 (m, 2H); Mass: [M+H]+ 761.50 (100%); HPLC:
98.07%.
Example 61: Preparation of 2,2-dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR, 13aRJ3bR)-5a,5b,8,8 J la-pentamethyl-3a-((2-methyl-l-(piperidin-l-yl)propan-2-yl) carbamoyl)- l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)butanoic acid:
Figure imgf000116_0002
Step 1: Synthesis of (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla-penta methyl-3a-((2-methyl-l -(piperidin-l -yl)propan-2-yl)carbamoyl)-l -(prop-1 -en-2-yl) icosahydro-lH-cyclopenta[a ]chrysen-9-yl acetate:
Figure imgf000116_0003
To a stirred solution of 2-methyl-l-(piperidin-l-yl)propan-2-amine (Intermediate 2, 0.800 g, 4.4 mmol, 2 eq) and triethylamine (2.7 mL, 11.0 mmol, 5.0 eq) in CH2C12 (50 ml) at 0 °C was added (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-(chlorocarbonyl)- 5a,5b,8,8,l la-pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (prepared as described in WO 2013/160810 A2, 1.2 g, 2.2 mmol, 1.0 eq) in CH2C12 (10 ml). The reaction mixture was allowed to stir at room temperature for overnight. The reaction mixture was diluted with water and extracted with CH2C12. The combined organic extracts were dried over Na2S04, filtered and evaporated under reduced pressure. The residue was purified by column chromatography by using 8% methanol: DCM as an eluent to afford the desired product (1.0 g, yield: 68.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 4.40-4.35 (q, 1H), 3.55 (t, 4H), 3.45 (bs, 4H), 2.76 (s, 1H), 2.57 (m, 1H), 2.39-2.10 (m, 13H), 1.96 (m, 3H), 1.91 (m, 2H), 1.69-1.05 (s, 16H), 0.95 (s, 3H), 0.87 (s, 3H), 0.84 (s, 3H), 0.82 (m, 12H), 0.34-0.29 (t, 2H) and 0.19-0.18 (q, 2H); Mass: [M]+ 694.39 (100%).
Step 2: Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-9-hydroxy-5a,5b,8,8, 11 a-pentamethyl-N -(2 -methyl- 1 -(piperidin-l -yl)propan-2-yl)-l -(prop-1 -en-2-yl)icosahydro- 3aH-cyclopenta[a]chrysene-3a-carboxamide:
Figure imgf000117_0001
To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8, l la-pentamethyl-3a-((2-methyl-l-(piperidin-l-yl)propan-2-yl)carbamoyl)-l-(prop-l-en-2-yl) icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (step 1, 1.0 g, 1.52 mmol, 1.0 eq) in THF (5 ml) and Methanol (5 ml) was added potassium carbonate (1.5 g, 10.64 mmol, 7.0 eq). The reaction mixture was stirred at room temperature for about 48 hours. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was filtered through a pad of celite and washed with CH2CI2. The filtrate was evaporated under reduced pressure and the crude was purified by column chromatography by using 1% methanol: dichloromethane as an eluent to afford the desired product (0.600 g, yield: 64.0%) as a white solid. 1H NMR (CDC13, 300 MHz): δ 4.28-4.26 (d, 1H), 3.55 (t, 4H), 3.44 (bs, 4H), 3.17 (d, 1H), 2.98 (q, 1H), 2.78-2.63 (m, 2H), 2.35-2.31 (m, 12H), 2.05 (bd, 1H), 1.95-1.87 (m, 3H), 1.61-1.25 (m, 19H), 0.93 (s, 3H), 0.86 (s, 9H), 0.77 (s, 3H), 0.65 (m, 3H), 0.34-0.29 (t, 2H) and 0.19-0.14 (q, 2H); Mass: [M]+ 652.51 (100%).
Step 3: Synthesis of 2,2-dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR, 13bR)-5a,5b,8, 8,11 a-pentamethyl- 3 a-((2 -methyl- 1 -(piperidin-l -yl)propan-2-yl)carbamoyl)-l - (prop-1 -en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)butanoic acid:
To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy- 5a,5b,8,8,l la-pentamethyl-N-(2-methyl-l-(piperidin-l-yl)propan-2-yl)-l-(prop-l-en-2-yl) icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide (step 2, 0.200 g, 0.328 mmol, 1.0 eq) and 2,2-dimethyl succinicanhydride (0.1312 g, 1.97 mmol, 4.0 eq) in toluene (10 ml) was added DMAP (0.080 g, 0.65 mmol, 2.0 eq). The reaction mixture was heated at 90°C for overnight. TLC indicated starting material was consumed and the desired product was observed. The mixture was concentrated under reduced pressure, cooled to 0°C, acidified to pH~5 with IN HC1 and extracted with CH2CI2. The combined organic extracts were washed with water, dried over Na2S04, filtered and evaporated under reduced pressure. The crude residue was purified by column chromatography by using 5% methanol: dichloromethane as an eluent to afford the desired product (0.050 g, yield: 19.0%) as a white solid. 1H NMR (CDC13, 300 MHz): δ 4.40-4.35 (q, 1H), 3.57-3.54 (m, 4H), 3.45 (t, 4H), 2.42-2.35 (m, 14H), 2.05 (bs, 1H), 1.87 (m, 2H), 1.65- 1.23 (m, 18H), 1.16 (m, 8H), 1.10-0.94 (m, 2H), 0.87 (s, 3H), 0.84-0.78 (m, 16H), 0.32 (t,lH) and 0.19 (t, 2H); Mass: [M]+ 723.49 (100%); HPLC: 92.91%.
The below examples 62-65 were prepared by the procedure similar to example-61 by using corresponding intermediates inpresence of suitable reagents, reactents and solvents at appropriate reaction conditions. The corresponding intermediates and the charctrization data are mentioned in the below table.
Figure imgf000118_0001
1H NMR (DMSO-d6, 300 MHz): δ
12.16 (s, 1H), 4.62 (s, 1H), 4.51
Λ (bs, 2H), 4.38 (m, 1H), 3.60-3.49
(m, 3H), 2.79 (m, 1H), 2.22-2.06 (m, 3H), 1.92 (m, 3H), 1.79-1.74
63 57
(m, 4H), 1.63-1.57 (m, 8H), 1.47- 1.29 (m, 11H), 1.29 (m, 8H), 1.23- 1.15 (m, 6H) and 0.92-0.19 (m, 16H); Mass: [M]+ 734.63 (60%); HPLC: 90.00%.
1H NMR (300 MHz, DMSO-d6): δ 11.86 (brs, 1H), 4.66 (s, 1H), 4.53
A. (s, 1H), 4.38-4.35 (m, 1H), 4.12
(m, 1H), 3.95 (m, 1H), 3.53-3.41
64 44
(m, 6H), 3.23 (s, 3H), 2.92- 2.22 (m, 10H), 1.98-1.06 (m, 37H), 0.94-0.76 (m, 16H); ESI Mass: 795.6 [M+H]+.
XH NMR (300 MHz, DMSO-d6): δ 0.77-0.80 (m, 9H), 0.85 (s, 3H), 0.93 (s, 5H), 1.06-1.16 (m, 12H), 1.20-1.35 (m, 11H), 1.40-1.59 (m, 4H), 1.64-1.75 (m, 6H), 1.83-1.94 (m, 5H), 2.11-2.17 (m, 1H), 2.22-
65 13 2.25 (m, 1H), 2.43 (m, 1H), 2.82-
Figure imgf000119_0001
2.91 (m, 5H), 3.07 (s, 5H), 3.45- 3.49 (m, 3H), 4.13 (bs, 1H), 4.34- 4.38 (m, 1H), 4.54 (s 1H), 4.67 (s, 1H), 10.04 (bs, 1H), 12.15 (bs,lH); Mass: [M+l]+ 764.63 (100%); HPLC Purity: 92.16%.
Example 66: synthesis of Synthesis of 5-(lR.3aS.5aR.5bR.7aR.9S.l laR.l lbR.13aR.13bR)-
3a-(l-(4-ethylpiperazin-l-yl)-2-methylpropan-2-yl)carbamoyl)-5a.5b.8.8J la-pentamethyl-l- (prop-l-en-2-yl)icosahvdro-lH-cvclopentaralchrvsen-9-yl)oxy)-3,3-dimethyl-5-oxopentanoic acid:
Figure imgf000120_0001
Step 1: Synthesis of (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-3a-(l-(4-ethylpiperazin -l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b,8,8,lla-pentamethyl-l-(prop-l-en-2-yl) icosahydro-l H-cyclopenta[a ]chrysen-9-yl acetate:
Figure imgf000120_0002
To a stirred solution of l-(4-ethylpiperazin-l-yl)-2-methylpropan-2-amine (intermediate 5, 5.20 g, 28.20 mmol, 2.0 eq) and triethylamine (10.0 mL, 70.00 mmol, 5.0 eq) in CH2C12 (70 ml) at 0 °C was added (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a- (chlorocarbonyl)-5a,5b,8,8, 1 la-pentamethyl- l-(l-methylcyclopropyl)icosahydro- 1H- cyclopenta[a]chrysen-9-yl acetate (prepared as described in WO 2013/160810 A2, 7.0 g, 14.10 mmol, 1.0 eq) in CH2C12 (10 ml). The reaction mixture was allowed to stir at room temperature for overnight. The reaction mixture was diluted with water and extracted with CH2C12. The combined organic extracts were dried over Na2S04, filtered and evaporated under reduced pressure. The residue was purified by column chromatography by using 3% methanol: DCM as an eluent gave the desired product (6.0 g, yield: 63.0%) as a solid. 1H NMR (DMSO-d6, 300 MHz): δ 6.71 (s, 1H), 4.66 (s, 1H), 4.54 (s, 1H), 4.27 (d, 1H), 2.99 (t, 2H), 2.57 (s, 2H), 2.31 (m, 5H), 2.12 (d, 1H), 1.88 (s, 3H), 1.80 (q, 2H), 1.62 (s, 3H), 1.57- 1.35 (m, 15H), 1.31(s, 9H), 1.28-1.10 (m, 3H), 1.05-0.91 (m, 6H), 0.91 (s, 9H), 0.75 (s, 3H) and 0.45 (q, 4H); Mass: [M-H]+ 666.56 (100%).
Step 2: Synthesis of (lR,3aS,5aR,5bR, 7aR,9S,l 1 aR,l lbR,l 3aR,l 3bR)-N-(l -(4-ethylpiperazin- l-yl)-2-methylpropan-2-yl)-9-hydroxy-5a,5b,8,8,lla-pentamethyl-l-(prop-l-en-2-yl) icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide:
Figure imgf000120_0003
To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(4- ethylpiperazin - l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b,8,8, 1 la-pentamethyl- 1 -(prop- 1- en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (step 1, 6.0 g, 8.8 mmol, 1.0 eq) in THF (48 ml) and Methanol (48 ml) was added potassium carbonate (12.5 g, 88.5 mmol, 10.0 eq). The reaction mixture was stirred at room temperature for about 48 hours. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was filtered through a pad of celite and washed with CH2CI2. The filtrate was evaporated under reduced pressure and the crude was purified by column chromatography by using 3% methanol: dichloromethane as an eluent gave the desired product (3.50 g, yield: 62.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 6.71 (s, 1H), 4.66 (s, 1H), 4.54 (s, 1H), 4.27 (d, 1H), 2.99 (t, 2H), 2.57 (s, 2H), 2.31 (m, 5H), 2.12 (d, 1H), 1.80 (q, 3H), 1.62 (s, 3H), 1.57-1.35 (m, 15H), 1.31(s, 9H), 1.28-1.10 (m, 2H), 1.05-0.91 (m, 7H), 0.91 (s, 9H), 0.75 (s, 3H) and 0.45 (q, 4H); Mass: [M-H]+ 624.58 (100%).
Step 3: Synthesis of 5-(lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-3a-(l-(4-ethyl piperazin-l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b,8,8,lla-pentamethyl-l-(prop-l-en-2- yl) icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)-3,3-dimethyl-5-oxopentanoic acid:
To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-N-(l-(4- ethylpiperazin- l-yl)-2-methylpropan-2-yl)-9-hydroxy-5a,5b, 8, 8,1 la-pentamethyl- l-(prop-l- en-2-yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide (step 2, 0.400 g, 0.64 mmol, 1.0 eq) and 4,4-dimethyldihydro-2H-pyran-2,6(3H)-dione (0.460 g, 3.2 mmol, 5.0 eq) in toluene (10 ml) was added DMAP (0.156 g, 1.20 mmol, 2.0 eq). The reaction mixture was heated at 90 °C for overnight. TLC indicated starting material was consumed and the desired product was observed. The mixture was concentrated under reduced pressure, cooled to 0 0 C, acidified to pH~5 with IN HC1 and extracted with CH2CI2. The combined organic extracts were washed with water, dried over Na2S04, filtered and evaporated under reduced pressure. The crude residue was purified by column chromatography by using 4% methanol and dichloromethane as an eluent gave the desired product (0.130 g, tield: 28.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 12.19 (s, 2H), 6.73 (s, 1H), 4.66 (d, 2H), 4.40 (q, 1H), 3.03 (q, 1H), 2.43-2.27 (m, 5H), 2.23 (d, 5H), 2.13 (d, 1H), 1.77 (d, 2H), 1.63 (d, 5H), 1.52-1.32 (m, 13H), 1.22 (d, 6H), 1.14 (t, 2H), 1.04 (m, 14H), 0.98 (m, 4H), and 0.86 (m, 15H); Mass: [M-H]+ 766.70 (100%); HPLC: 93.89%.
The below example 67 were prepared by the procedure similar to example-66 by using corresponding intermediates inpresence of suitable reagents, reactents and solvents at appropriate reaction conditions. The corresponding intermediates and the charctrization data are mentioned in the below table. Inter
Ex
media Structure Characterization data no.
te no
1H NMR (300 MHz, DMSO-d6): δ 0.79-0.80 (m, 9H ), 0.85 (s, 3H), 0.93 (s, 5H), 1.04-1.16 (m, 11H), 1.21-1.40 (m, 9H), 1.44-1.59 (m, 4H), 1.64-1.75 (m, 9H), 1.86-1.92 (m, 2H), 2.11-2.17 (m, 1H), 2.22-
67 13 2.38 (m, 8H), 2.83-2.93 (m, 5H),
3.07 (s, 4H), 3.42-3.49 (m, 3H), 4.12 (bs, 1H), 4.35-4.39 (m, 1H), 4.54 (s 1H), 4.67 (s, 1H), 9.75 (bs, 1H), 12.00 (bs, 1H); Mass: [M+l]+ 778.66 (100%); HPLC: 96.44%.
Example 68: 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-((2-methyl-l-(pyrrolidin-l-yl)propan-2-yl) carbamoyl)- l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)carbonyl) cyclobutane-l-carboxylic acid:
Figure imgf000122_0001
Step 1: Synthesis (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla-penta methyl-3a-((2-methyl-l -(pyrrolidin-l -yl)propan-2-yl)carbamoyl)-l -(prop-1 -en-2-yl) icosahydro-lH-cyclopenta[a ]chrysen-9-yl acetate:
Figure imgf000122_0002
To a stirred solution of 2-methyl-l-(pyrrolidin-l-yl)propan-2-amine (Intermediate 3, 2.2 g, 16.00 mmol, 2.0 eq) and triethylamine (6.0 mL, 40.0 mmol, 5.0 eq) in CH2C12 (20 ml) at 0 °C was added (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-(chlorocarbonyl)- 5a,5b,8,8,l la-pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (prepared as described in WO 2013/160810 A2, 4.0 g, 80.0 mmol, 1.0 eq) in CH2C12 (20 ml). The reaction mixture was allowed to stir at room temperature for overnight. The reaction mixture was diluted with water and extracted with CH2C12. The combined organic extracts were dried over Na2S04, filtered and evaporated under reduced pressure. The residue was purified by column chromatography by using 3% methanol: DCM as an eluent to afford the desired product (2.5 g, yield: 51.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 6.79 (s, 1H), 4.65 (d, 2H), 4.38 (q, 1H), 3.55 (t, 4H), 3.44 (bs, 3H), 3.17 (d, 1H), 2.98 (q, 1H), 2.78-2.63 (m, 2H), 2.35-2.31 (m, 12H), 2.05 (bd, 1H), 1.99 (s, 3H), 1.95-1.87 (m, 3H), 1.61- 1.25 (m, 14H), 0.93 (s, 3H), 0.86 (s, 9H), 0.77 (s, 3H) and 0.65 (m, 3H); Mass: [M]+ 623.51 (100%).
Step 2: Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-9-hydroxy-5a,5b,8,8, 11 a-pentamethyl-N -(2 -methyl- 1 -(pyrrolidin-l -yl)propan-2-yl)-l -(prop-1 -en-2-yl)icosahydro- 3aH-cyclopenta[a ]chrysene-3a-carboxamide:
Figure imgf000123_0001
To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8, 1 la-pentamethyl-3a-((2-methyl-l-(pyrrolidin- l-yl)propan-2-yl)carbamoyl)- l-(prop- 1-en- 2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (step 1, 2.2 g, 40.0 mmol, 1.0 eq) in THF (18 ml) and Methanol (18 ml) was added potassium carbonate (4.8 g, 35.0 mmol, 10.0 eq). The reaction mixture was stirred at room temperature for about 48 hours. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was filtered through a pad of celite and washed with CH2C12. The filtrate was evaporated under reduced pressure and the crude was purified by column chromatography by using 3% methanol: dichloromethane as an eluent to afford the desired product (1.5 g, yield: 72.0%) as a white solid. 1H NMR (CDC13, 300 MHz): δ 6.82 (s, 1H), 4.65 (d, 2H), 3.55 (t, 4H), 3.44 (bs, 4H), 3.17 (d, 1H), 2.98 (q, 1H), 2.78-2.63 (m, 2H), 2.35-2.31 (m, 12H), 2.05 (bd, 1H), 1.95-1.87 (m, 3H), 1.61-1.25 (m, 15H), 0.93 (s, 3H), 0.86 (s, 9H), 0.77 (s, 3H) and 0.65 (m, 3H); Mass: [M]+ 581.53 (100%).
Step 3: Synthesis of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-5a,5b, 8,8,11 a-pentamethyl- 3 a-((2 -methyl- 1 -(pyrrolidin-l -yl)propan-2-yl)carbamoyl)-l -(prop-1 -en- 2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-dimethylcyclobutane-l,3- dicarboxylate:
Figure imgf000124_0001
To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy- 5a,5b,8,8,l la-pentamethyl-N-(2-methyl-l-(pyrrolidin-l-yl)propan-2-yl)-l-(prop-l-en-2- yl)icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide (step 2, 1.5 g, 2.5 mmol, 1.0 eq) in DCM (30 ml) then added (lS,3R)-3-((benzyloxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid (prepared as described in WO 2011/007230 A2, 1.0 g, 3.8 mmol, 1.5 eq) and the reaction mixture was cooled to 0 °C then added DCC (1.0 g, 5.0 mmol, 2.0 eq) followed by DMAP (0.063 g, 0.5 mmol, 0.2 eq). The reaction mixture was raised to room temperature and stirred for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was evaporated under reduced pressure and diluted with water. The aqueous layer was extracted with CH2CI2. The organic layer was washed with water and brine solution. The combined organic layers were dried over Na2S04, filtered and evaporated under reduced pressure. The crude residue was purified by column chromatography by using 4% MeOH and DCM as an eluent to obtain the desired product (1.2 g, yield: 58.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 7.36 (s, 5H), 6.79 (s, 1H), 5.48 (q, 2H), 4.65 (d, 2H), 4.36 (t, 1H), 3.00 (t, 2H), 2.82 (t, 3H), 2.34 (t, 2H), 2.14 (s, 1H), 1.90-1.83 (m, 6H), 1.63 (s, 3H), 1.57-1.47 (m, 4H), 1.43-1.28 (m, 22H), 1.18-0.98 (m, 4H) and 0.92-0.80 (m, 22H); Mass: [M]+ 825.64 (100%).
Step 4: Synthesis of (lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR, 13bR)-5a,5b,8,8 la^entamethyl-3a-((2-methyl-l-(pyrrolidin-l-yl)propan-2-yl)carbamoyl)-
1- (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l- carboxylic acid:
To a solution of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b,
8,8, 1 la-pentamethyl-3a-((2-methyl-l-(pyrrolidin- l-yl)propan-2-yl)carbamoyl)- l-(prop- 1-en-
2- yl)icosahydro- 1 H-cyclopenta[a] chry sen-9-yl) ( 1 R,3 S )-2,2-dimethylcyclobutane- 1,3- dicarboxylate (step 3, 0. 600 g, 0.72 mmol, 1.0 eq) in THF (20 mL) then added triethylamine (0.2 mL, 1.4 mmol, 2.0 eq) and triethylsiline (0.128 mL, 1.0 mmol, 1.5 eq) followed by palladium acetate (0.100 g). The reaction mixture was refluxed for overnight. The reaction mixture was filtered through a pad of celite, was washed with MeOH and DCM. The filtrate was evaporated under reduced pressure and the crude residue was purified by column chromatography by using 8% methanol: DCM as an eluent to afford the desired compound (0.180g, yield: 38.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 12.10 (s, 1H), 6.99 (s, 1H), 4.65 (d, 2H), 4.36 (t, 1H), 3.00 (t, 2H), 2.82 (t, 3H), 2.34 (t, 2H), 2.14 (s, 1H), 1.90-1.83 (m, 6H), 1.63 (s, 3H), 1.57-1.47 (m, 4H), 1.43-1.28 (m, 22H), 1.18-0.98 (m, 4H) and 0.92-0.80 (m, 22H); Mass: [M]+ 735.63 (100%); HPLC: 93.64%.
The below examples 69-90 were prepared by the procedure similar to example-68 by using corresponding intermediates inpresence of suitable reagents, reactents and solvents at appropriate reaction conditions. The corresponding intermediates and the charctrization data are mentioned in the below table.
Figure imgf000125_0001
Figure imgf000126_0001
Figure imgf000127_0001
1H NMR (300 MHz, CDC13): δ
7.41-7.28 (m, 5H), 6.77 (s, IH), 4.72 (s, IH), 4.57 (s, IH), 4.44-
77 10 4.39 (m, IH), 3.11-1.97 (m, 28H),
1.67-1.25 (m, 22H), 1.13 (t, J = 6.9 Hz, 3H), 1.05-0.75 (m, 22H); ES Mass: 869.66 [M+H]+.
1H NMR (300 MHz, CDC13): δ 12.04 (brs, IH), 7.20 (brs, IH), 4.64 (s, IH), 4.53 (s, IH), 4.36- 4.30 (m, IH), 3.96 (m, IH), 3.01-
78 54
HO 2.98 (m, IH), 2.82-2.75 (m, 2H),
2.61-2.13 (m, 5H), 1.89-1.16 (m, 31H), 1.11-0.77 (m, 27H); ES Mass: 721.53 [M+H]+.
1H NMR (300 MHz, DMSO-d6): δ 12.04 (brs, IH), 7.11 (m, IH), 4.64 (s, IH), 4.52 (s, IH), 4.36- 4.31 (m, IH), 4.06 (m, IH), 3.02-
79 56 2.99 (m, IH), 2.82-2.72 (m, 2H),
Figure imgf000128_0001
2.64-2.15 (m, 8H), 1.89-1.78 (m, 3H), 1.62-1.01 (m, 41H), 0.92- 0.80 (m, 19H); ES Mass: 777.6 [M+H]+.
Figure imgf000129_0001
1H NMR (DMSO-d6, 300 MHz): δ
0.81-0.86 (m, 12H), 0.87-0.94 (m, 13H), 1.03-1.15 (m, 5H), 1.26-
1.1.53 (m, 9H), 1.58-1.65 (m, 6H), 1.75-1.89 (m, 5H), 2.06 (bs, 3H), 2.24-2.31 (m, 6H), 2.72-2.82 (m,
83 46
HOOCT 3H), 2.83-2.92 (m, 2H), 3.47 (bs,
2H), 3.72-3.79 (m, 1H), 4.12-4.16 (m, 1H), 4.31-4.36 (m, 2H), 4.54 (bs, 1H), 4.67 (bs, 1H), 5.77 (s, 1H), 12.05 (bs, 1H); Mass: [M]+
772.54 (M + H)+; HPLC: 92.13%.
1H NMR (DMSO-d6, 300 MHz): δ
Λ 4.67 (s, 1H), 4.53 (s, 1H), 4.37- 4.28 (m, 1H), 4.21-4.12 (m, 1H), 3.58-3.48 (m, 6H), 2.93-2.78 (m,
84 16
HOOCT 4H), 2.42-2.22 (m, 5H), 2.11-1.02
(m, 33H), 0.92-0.72 (m, 21H); Mass: [M+H]+ 763.64 (100%); HPLC: 90.0%.
1H NMR (DMSO-d6, 300 MHz): δ 0.75 (s, 1H), 0.81 (m, 9H), 0.86 (m, 3H), 0.88 (m, 3H), 0.90 (m, 3H), 0.94 (m, 3H), 1.06-1.14 (m, 3H), 1.23-1.26 (m, 7H), 1.33-1.59 (m, 14H), 1.65 (m, 6H), 1.83-2.06
85 43 (m, 5H), 2.76-2.78 (m, 2H), 2.82-
Figure imgf000130_0001
2.97 (m, 2H), 4.12-4.19 (m, 1H),
4.26-4.34 (m, 2H), 4.55 (s, 1H), 4.69 (s, 1H), 6.23 (s, 1H), 7.43 (s, 1H), 7.61 (s, 1H), 12.10 (s, 1H); Mass: [M]+ 744.61 (M + H)+; HPLC: 91.10%.
Figure imgf000131_0001
1H NMR (300 MHz, CDC13): δ
4.72 (s, 1H), 4.57 (s, 1H), 4.46-
Λ. 4.36 (m, 1H), 4.25-4.23 (m, 1H),
3.82-3.78 (m, 1H), 2.97-2.74 (m,
90 42 6H), 2.63-2.56 (m, 1H), 2.37-2.32
HO (m, 1H), 2.20-1.96 (m, 12H),
1.42-1.20 (m, 20H), 1.14-1.12 (m, 3H), 1.06-0.84 (m, 24H), 0.67 (s, 3H), 0.64 (s, 3H); HPLC: 92.4%.
Example 91: 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-((2-(pyrrolidin-l-yl methyl)- lH-pyrrol-l-yl)methyl)pyrrolidine-l-carbonyl)icosahydro-lH-cyclopentaralchrysen- 9-yl)oxy)carbonyl)cyclobutane- 1-carboxylic acid:
Figure imgf000132_0001
Step 1: Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla-penta methyl-1 -(prop-1 -en-2-yl)-3a-((S)-2-((2-(pyrrolidin-l -ylmethyl)-lH-pyrrol-l-yl)methyl) pyrrolidine-1 -carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate:
Figure imgf000132_0002
To a stirred solution of (S)-2-(pyrrolidin-l-ylmethyl)-l-(pyrrolidin-2-ylmethyl)-lH- pyrrole hydrochloride (Intermediate 36, 0.75 g, 2.9 mmol) in DCM (10 ml) and NEt3 (4.5 ml, 30.1 mmol), was added a solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a- (chlorocarbonyl)-5a,5b,8,8,l la-pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a] chrysen-9-yl acetate (prepared as described in WO 2013/160810 A2, 1.6 g, 3.05 mmol) in DCM (20 ml) at 0 °C and allowed to stir at room temperature for about 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, brine and dried over Na2S04, and the solvent was evaporated and purified by silica gel column (elution 2% MeOH in DCM) to afford the title compound (1.3 g, yield: 65%) as an off white solid. 1H NMR (300 MHz, CDC13): δ 6.67 (m, 1H), 6.25 (m, 1H), 6.09 (m, 1H), 4.70 (s, 1H), 4.59 (s, 1H), 4.52-4.48 (m, 1H), 4.42-4.28 (m, 2H), 4.22-4.19 (m, 1H), 3.92-3.52 (m, 3H), 3.05-2.82 (m, 5H), 2.32-1.16 (m, 33H), 0.96-0.75 (m, 21H). 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- ( (2-(pyrrolidin-l -ylmethyl)-! -pyrrol-1 -yl)methyl)pyrrolidin-l -yl)methanone:
Figure imgf000133_0001
To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8, 1 la-pentamethyl- l-(prop- l-en-2-yl)-3a-((S)-2-((2-(pyrrolidin- 1 -ylmethyl)- lH-pyrrol- 1-yl) methyl)pyrrolidine-l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (step 1, 1.3 g, 1.82 mmol) in MeOH: THF (6 ml: 6 ml), was added 4N NaOH (6 ml) at 0 °C and allowed to stir at room temperature for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure. The reaction mixture was diluted with DCM and washed with water, brine and dried over Na2S04. The solvent was evaporated and the resulting solid was taken in hexane and stirred for one hour and filtered to afford the title compound (1.15 g, yield: 94%) as a white solid. 1H NMR (300 MHz, DMSO- d6): δ 6.66 (m, 1H), 5.89-5.85 (m, 2H), 4.65 (s, 1H), 4.54 (s, 1H), 4.28-4.13 (m, 3H), 3.88- 3.84 (m, 1H), 3.64-3.49 (m, 3H), 2.96-2.93 (m, 4H), 2.43-1.02 (m, 35H), 0.93-0.64 (m, 18H). Step 3: Synthesis of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-5a,5b, 8,8,11 a-pentamethyl-1 -(prop-1 -en-2-yl)-3a-((S)-2-((2-(pyrrolidin-l -ylmethyl)-! H-pyrrol-1 - yl)methyl)pyrrolidine-l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2- dimethylcyclobutane-l,3-dicarboxylate:
Figure imgf000133_0002
To a stirred solution of ((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy -5a,5b,8,8,l la-pentamethyl- l-(prop-l-en-2-yl)icosahydro-3aH-cyclopenta[a]chrysen-3a-yl) ((S)-2-((2-(pyrrolidin-l-ylmethyl)-lH-pyrrol-l-yl)methyl)pyrrolidin-l-yl)methanone (step 2, 1.15 g, 1.7 mmol) and (lS,3R)-3-((benzyloxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid (prepared as described in WO 2011/007230 A2, 0.67 g, 2.5 mmol) and
DMAP (0.02 g, cat) in DCM (20 ml), was slowly added DCC (0.71 g, 3.0 mmol) in DCM (8 ml) at 0 °C and allowed to stir at room temperature for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHC03 solution, brine and dried over Na2S04. The solvent was evaporated and to the resulting solid, was added DCM (10 ml) and stirred for one hour and filtered. The filtrate was concentrated under reduced pressure to afford the title compound as a white solid (1.05 g, yield: 67%). 1H MR (300 MHz, CDC13): δ 7.34 (m, 5H), 6.68 (m, 1H), 6.19-6.08 (m, 2H), 5.12 (ABq, J = 12.3 Hz, 2H), 4.72 (s, 1H), 4.59 (s, 1H), 4.46-3.42 (m, 7H), 3.05-2.58 (m, 6H), 2.35-1.13 (m, 39H), 0.99-0.75 (m, 21H).
Step 4: Synthesis of (lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR, 13bR)-5a,5b,8,8,lla-pentamethyl-l-(prop-l-en-2-yl)-3a-((S)-2-((2-(pyrrolidin-l-ylmethyl)- lH^yrrol-l-yl)methyl)pyrrolidine-l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) oxy)carbonyl)cyclobutane-l -carboxylic acid:
To a stirred solution of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,
13bR)-5a,5b,8,8,l la-pentamethyl-l-(prop-l-en-2-yl)-3a-((S)-2-((2-(pyrrolidin-l-ylmethyl)- lH-pyrrol-l-yl)methyl)pyrrolidine-l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-dimethylcyclobutane-l,3-dicarboxylate (step 3, 0.35 g, 0.3 mmol) in THF (8 ml), were added N(Et)3 (0.2 ml, 1.5 mmol), triethyl silane (0.2 ml, 0.45 mmol) and Pd(OAc)2 (0.02 g, cat). The reaction mixture was heated to 70 °C and continued for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc and washed with water, saturated NaHC03 solution, brine and dried over Na2S04. The solvent was evaporated under reduced pressure and purified by silica gel column (elution 5% MeOH in DCM) to afford the title compound as a white solid (0.1 g, yield: 32.2%). 1H NMR (300 MHz, DMSO-d6): δ 12.15 (brs, 1H), 6.89 (m, 1H), 6.29 (m, 1H), 6.03 (m, 1H), 4.65 (s, 1H), 4.56 (s, 1H), 4.39-4.32 (m, 1H), 4.21-4.05 (m, 2H), 3.24- 3.46 (m, 3H), 3.12-2.71 (m, 6H), 2.37-1.09 (m, 40H), 0.95-0.81 (m, 21H); ES Mass: 826.7 [M+H]+; HPLC: 90.86%. Example 92: 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-((2-methyl-lH-pyrrol-l-yl)methyl) pyrrolidine- l-carbonyl)-l-(prop-l-en-2-yl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy) carbonvDcyclobutane-l -carboxylic acid:
Figure imgf000135_0001
To a stirred solution of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR) -5a,5b,8,8,l la-pentamethyl-l-(prop-l-en-2-yl)-3a-((S)-2-((2-(pyrrolidin-l-ylme l)-lH- pyrrol-l-yl)methyl)pyrrolidine-l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-dimethylcyclobutane-l,3-dicarboxylate (Example 91-step 3, 0.35 g, 0.3 mmol) in THF (8 ml), were added N(Et)3 (0.2 ml, 1.5 mmol), triethyl silane (0.2 ml, 0.45 mmol) and Pd(OAc)2 (0.02 g, cat). The reaction mixture was heated to 70 °C and continued for about 18 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc and washed with water, saturated NaHC03 solution, brine and dried over Na2S04. The solvent was evaporated under reduced pressure and purified by silica gel column (elution 2% MeOH in DCM) to afford the title compound (0.12 g, yield: 42%) as a white solid. 1H MR (300 MHz, CDC13): δ 6.57-6.56 (m, 1H), 6.04-6.02 (m, 1H), 5.85 (m, 1H), 4.75 (s, 1H), 4.59 (s, 1H), 4.45-4.43 (m, 2H), 4.19-4.15 (m, 1H), 3.58 (m, 1H), 3.42 (m, 1H), 2.96-2.71 (m, 6H), 2.26 (s, 3H), 2.04-1.11 (m, 30H), 1.06-0.76 (m, 22H); ES Mass: 757.68 [M+H]+; HPLC: 92.60%.
Example 93 : 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-l-(prop-l-en-2-vn-3a-((S)-2-(pyrrolidin-l-ylmethvn pyrrolidine- l-carbonyl)icosahydro-lH-cyclopenta[alchrysen-9-yl)oxy)carbonyl)cyclobutane- 1-carboxylic acid and
Example 94: Preparation of (lR.3S)-3-((((lR.3aS.5aR.5bRJaR.9S.l laR.l lbR.13aR.13bR)- 3a-((S)-2-((lH-pyrrol-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl-l- (prop-l-en-2-yl)icosahydro-lH-cyclopenta[alchrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclo butane- 1-carboxylic acid;
Figure imgf000135_0002
Step 1: Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla-penta methyl-1 -(prop-1 -en-2-yl)-3a-((S )-2-(pyrrolidin-l -ylmethyl)pyrrolidine-l -carbonyl) icosahydro-lH-cyclopenta[a] chrysen-9-yl acetate:
Figure imgf000136_0001
To a stirred solution of (S)-l-(pyrrolidin-2-ylmethyl)pyrrolidine (Intermediate 39, 0.98 mL, 6.01 mmol, 1.5 eq) and triethylamine (1.67 mL, 12.03 mmol, 3.0 eq) in CH2C12 (50 ml) at 0 °C was added (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-(chlorocarbonyl) -5a,5b,8,8,l la-pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (prepared as described in WO 2013/160810 A2, 2.0 g, 4.01 mmol, 1.0 eq) in CH2C12 (10 ml). The reaction mixture was allowed to stir at room temperature for overnight. The reaction mixture was diluted with water and extracted with CH2C12. The combined organic extracts were dried over Na2S04, filtered and evaporated under reduced pressure. The residue was purified by column chromatography by using 3% methanol: DCM as an eluent gave the desired product (1.2 g, yield: 48.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 4.65 (s, 1H), 4.53 (s, 1H), 4.27 (d, 1H), 4.14 (m, 1H), 3.50 (m, 1H), 2.99-2.86 (m, 3H), 2.27 (m, 2H), 1.99 (s, 3H), 1.96-1.63 (m, 9H), 1.58 (s, 3H), 1.53-1.23 (m, 21H) and 0.93-0.78 (m, 20H); Mass: [M]+ 635.72 (100%).
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- (pyrrolidin-l -ylmethyl)pyrrolidin-l -yl)methanone.
Figure imgf000136_0002
To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8, l la-pentamethyl-l-(prop-l-en-2-yl)-3a-((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidine-l-carbonyl) icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (step 1, 1.2 g, 1.89 mmol, 1.0 eq) in THF (10 ml), Methanol (10 ml) and water (15 mL) was added lithium hydroxide (0.85 g, 18.92 mmol, 10.0 eq). The reaction mixture was stirred at room temperature for about 12 hours. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was diluted with water and extracted in DCM. The organic layer was dried with Na2S04, and filtrate was evaporated under reduced pressure and the crude was purified by column chromatography by using 6% methanol: dichloromethane as an eluent gave the desired product (0.600 g, yield: 54.0%) as a white solid. 1H NMR (CDC13, 300 MHz): δ 4.65 (s, 1H), 4.53 (s, 1H), 4.27 (d, 1H), 4.14 (m, 1H), 3.50 (m, 2H), 2.99-2.86 (m, 3H), 2.27 (m,
2H), 1.96-1.63 (m, 9H), 1.58 (s, 3H), 1.53-1.23 (m, 21H), 1.13-1.05 (m, 2H), 0.91 (s, 3H), 0.86 (s, 9H), 0.76 (s, 3H) and 0.64 (s, 3H); Mass: [M]+ 593.68 (100%).
Step 3: Synthesis of 1-benzyl 3-(lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8, lla-pentamethyl-l-(prop-l-en-2-yl)-3a-((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidine-l-carbonyl) icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-dimethylcyclobutane-l,3- dicarboxylate:
Figure imgf000137_0001
To a stirred solution of ((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy -5a,5b,8,8,l la-pentamethyl-l-(prop-l-en-2-yl)icosahydro-3aH-cyclopenta[a]chrysen-3a-yl) ((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidin-l-yl)methanone (step 2, 0.600 g, 1.01 mmol, 1.0 eq) in DCM (50 ml) then added (lS,3R)-3-((benzyloxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid (prepared as described in WO 2011/007230 A2, 0.39 g, 1.52 mmol, 1.5 eq) this reaction mixture was cooled to 0 °C then added DCC (0.41 g, 2.02 mmol, 2.0 eq) followed by DMAP (0.02 g, 0.20 mmol, 0.2 eq). The reaction mixture was raised to room temperature and stirred for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was evaporated under reduced pressure and diluted with water. The aqueous layer was extracted with CH2CI2. The organic year was washed with water and brine solution. The combined organic layers were dried over Na2S04, filtered and evaporated under reduced pressure. The crude residue was purified by column chromatography by using 5% MeOH and DCM as an eluent to obtain the desired product (0.450 g, yield: 56.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 7.36-7.32 (m, 5H), 5.14 (q, 2H), 4.66 (s, 1H), 4.53 (s, 1H), 3.36 (m, 1H), 4.14 (bs, 1H), 3.50 (s, 1H), 2.88- 2.76 (m, 4H), 2.34-2.23 (m, 6H), 1.91-1.85 (m, 7H), 1.67-1.64 (m, 9H), 1.58-1.32 (m, 11H), 1.29 (m, 6H), 1.17 (m, 3H), 0.93 (m, 9H) and 0.75 (m, 13H); Mass: [M]+ 837.69 (100%).
Step 4: Synthesis of (lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR, 13bR)-5a,5b,8,8,lla-pentamethyl-l-(prop-l-en-2-yl)-3a-((S)-2-(pyrrolidin-l-ylmethyl) pyrrolidine-1 -carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane- 1 -carboxylic acid and
Synthesis of (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-3a-((S)-2-((lH- pyrrol-1 -yl )methy I pyrrolidine -1 -carbonyl)-5a,5b, 8,8,1 la-pentamethyl-1 -(prop-1 -en-2-yl) icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid
To a solution of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8, 1 la-pentamethyl- l-(prop- l-en-2-yl)-3a-((S)-2-(pyrrolidin- l-ylmethyl)pyrrolidine- 1- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-dimethylcyclobutane-l,3- dicarboxylate (step 3, 0. 450 g, 0.53 mmol, 1.0 eq) in THF (30 niL) then added triethylamine (0.7 niL, 1.06 mmol, 2.0 eq) and triethylsiline (0.1 mL, 0.64 mmol, 1.2 eq) followed by palladium acetate (0.050 g). The reaction mixture was refluxed for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was filtered through a pad of celite and was washed with MeOH and DCM. The filtrate was evaporated under reduced pressure, the crude residue was purified by column chromatography by using 10% methanol: DCM as an eluent to give the desired compound Example 93 (0.120 g, yield: 60.0%) as a white solid and Example 94 (0.110 g, yield: 60.0%) as a white solid.
Example 93: 1H NMR (DMSO-d6, 300 MHz): δ 12.16 (s, 1H), 4.66 (s, 1H), 4.53 (s, 1H), 3.36 (m, 1H), 4.14 (bs, 1H), 3.50 (s, 1H), 2.88-2.76 (m, 4H), 2.34-2.23 (m, 6H), 1.91-1.85 (m, 7H), 1.67-1.64 (m, 9H), 1.58-1.32 (m, 11H), 1.29 (m, 6H), 1.17 (m, 3H), 0.93 (m, 9H) and 0.75 (m, 13H); Mass: [M]+ 747.54 (100%); HPLC: 97.59%.
Example 94: 1H NMR (DMSO-d6, 300 MHz): δ 12.17 (s, 1H), 6.67 (s, 2H), 5.99 (s, 2H), 4.70 (s, 1H), 4.55 (s, 1H), 3.36 (m, 1H), 4.36 (t, 1H), 4.19 (m, 1H), 4.04-3.94 (m, 2H), 3.46 (m, 1H), 2.96 (m, 2H), 2.82 (m, 2H), 2.34-2.17 (m, 1H), 2.11-1.83 (m, 2H), 1.77-1.23 (m, 22H), 1.19-1.00 (m, 3H) and 0.90-0.74 (m, 25H); Mass: [M]+ 743.54 (100%); HPLC: 92.00%. Example 95: Preparation of (lS,3R)-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-(pyrrolidin-l-ylmethyl) pyrrolidine- l-carbonyl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)carbonyl)cyclobutane- 1-carboxylic acid:
Figure imgf000138_0001
Step 1: Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla-penta methyl-1 -(prop-1 -en-2-yl)-3a-((S)-2-(pyrrolidin-l -ylmethyl)pyrrolidine-l -carbonyl) icosahydro-lH-cyclopenta[a] chrysen-9-yl acetate:
Figure imgf000139_0001
To a stirred solution of (S)-l-(pyrrolidin-2-ylmethyl)pyrrolidine (Intermediate 39, 0.61 g, 4.01mmol, 1.0 eq) and triethylamine (1.7 mL, 11.88 mmol, 3.0 eq) in CH2C12 (15 ml) at O °C was added (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-(chlorocarbonyl)-5a, 5b,8,8,l la-pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (prepared as described in WO 2013/160810 A2, 2.0 g, 4.01 mmol, 1.0 eq) in CH2C12 (10 ml). The reaction mixture was allowed to stir at room temperature for overnight. The reaction mixture was diluted with water and extracted with CH2C12. The combined organic extracts were dried over Na2S04, filtered and evaporated under reduced pressure. The residue was purified by column chromatography by using 3% methanol: DCM as an eluent gave the desired product (1.2 g, yield: 60.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 4.65 (s, 1H), 4.53 (s, 1H), 4.27 (d, 1H), 4.14 (m, 1H), 3.50 (m, 1H), 2.99-2.86 (m, 3H), 2.27 (m, 2H), 1.99 (s, 3H), 1.96-1.63 (m, 9H), 1.58 (s, 3H), 1.53-1.23 (m, 21H) and 0.93-0.78 (m, 20H); Mass: [M]+ 635.72 (100%).
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- (pyrrolidin-l -ylmethyl)pyrrolidin-l -yl)methanone:
Figure imgf000139_0002
To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8, l la-pentamethyl-l-(prop-l-en-2-yl)-3a-((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidine-l-carbonyl) icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (step 1, 1.2 g, 1.89 mmol, 1.0 eq) in THF (10 ml) and Methanol (10 ml) was added potassium carbonate (1.99 g, 15.14mmol, 8 eq). The reaction mixture was stirred at room temperature for about 12 hours. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was evaporated through reduced pressure then diluted with water and extracted in DCM. The organic layer was dride with Na2S04, and filtrate was evaporated under reduced pressure and the crude was purified by column chromatography by using 6% methanol: dichloromethane as an eluent gave the desired product (0.600 g, yield: 54.0%) as a white solid. 1H NMR (CDCI3, 300 MHz): δ 4.65 (s, 1H), 4.53 (s, 1H), 4.27 (d, 1H), 4.14 (m, 1H), 3.50 (m, 2H), 2.99-2.86 (m, 3H), 2.27 (m, 2H), 1.96-1.63 (m, 9H), 1.58 (s, 3H), 1.53-1.23 (m, 21H), 1.13- 1.05 (m, 2H), 0.91 (s, 3H), 0.86 (s, 9H), 0.76 (s, 3H) and 0.64 (s, 3H); Mass: [M]+ 593.68 (100%).
Step 3: Synthesis of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8, lla-pentamethyl-l-(prop-l-en-2-yl)-3a-((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidine-l-carbonyl) icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lS,3R)-2,2-dimethylcyclobutane-l,3- dicarboxylate:
Figure imgf000140_0001
To a stirred solution of ((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy
-5a,5b,8,8,l la-pentamethyl-l-(prop-l-en-2-yl)icosahydro-3aH-cyclopenta[a]chrysen-3a-yl) ((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidin-l-yl)methanone (step 2, 0.600 g, 1.01 mmol, 1.0 eq) in DMF (20 ml) then added (lR,3S)-3-((benzyloxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid (prepared as described in WO 2014/105926 Al, 0.39 g, 1.52 mmol, 1.5 eq). The reaction mixture was cooled to 0 °C then added EDCI (0.38 g, 2.02 mmol, 2.0 eq) followed by DMAP (0.024 g, 0.20 mmol, 0.2 eq). The reaction mixture was raised to room temperature stirred for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was quenching with ice cold water, solid was precipitated out then filtered through Buchner funnel, solid compound was dissolved in DCM and washed with water, sodium bicarbonate solution followed by brine solution. The combined organic layers were dried over Na2S04, filtered and evaporated under reduced pressure. The crude residue was purified by column chromatography by using 5% MeOH and DCM as an eluent to obtain the desired product (0.450 g, yield: 56.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 7.36-7.32 (m, 5H), 5.14 (q, 2H), 4.66 (s, 1H), 4.53 (s, 1H), 3.36 (m, 1H), 4.14 (bs, 1H), 3.50 (s, 1H), 2.88-2.76 (m, 4H), 2.34-2.23 (m, 6H), 1.91-1.85 (m, 7H), 1.67-1.64 (m, 9H), 1.58-1.32 (m, 11H), 1.29 (m, 6H), 1.17 (m, 3H), 0.93 (m, 9H) and 0.75 (m, 13H); Mass: [M]+ 837.69 (100%).
Step 4: Synthesis of (lS,3R)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR, 13bR)-5a,5b,8,8,lla-pentamethyl-l-(prop-l-en-2-yl)-3a-((S)-2-(pyrrolidin-l-ylmethyl) pyrrolidine-1 -carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane- 1 -carboxylic acid: To a solution of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8, 1 la-pentamethyl- l-(prop- l-en-2-yl)-3a-((S)-2-(pyrrolidin- l-ylmethyl)pyrrolidine- 1- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lS,3R)-2,2-dimethylcyclobutane-l,3- dicarboxylate (step 3, 0. 450 g, 0.53 mmol, 1.0 eq) in Ethyl acetate (15 mL) and Methanol (15ml) then added by Pd/C (0.100 g) followed by added ammonium formate (0.16 g, 2.06 mmol) under nitrogen atmosphere. The reaction mixture was stirred at room temperature for about 4 hours. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was filtered through a pad of celite and was washed with MeOH and DCM. The filtrate was evaporated under reduced pressure, the crude residue was purified by column chromatography by using 10% methanol: DCM as an eluent gave the desired compound (0.120 g, yield: 60.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 12.13 (bs, 1H), 4.66 (s, 1H), 4.53 (s, 1H), 4.31-4.35 (m, 1H), 4.13 (bs, 1H), 3.51 (bs, 1H), 3.40 (bs, 1H), 2.71-2.91 (m, 4H), 2.23-2.43 (m, 9H), 1.73-1.79 (m, 6H), 1.59-1.65 (m, 11H), 1.40-1.49 (m, 5H), 1.23-1.36 (m, 12H), 1.04-1.15 (m, 5H), 0.86-0.99 (m, 10H) and 0.79-0.81 (m, 9H); Mass: [M]+ 747.54 (100%); HPLC: 89.5%.
Example 96: Preparation of (lR,3S)-3-((((lS,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- 1 -isopropyl-5 a,5b, 8 ,8 , 11 a-pentamethyl-3 a-((2-methyl- 1 -(pyrrolidin- 1 -yl)propan-2-yl) carbamoyl)icosahydro-lH-cyclopentaralchrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclo butane- 1-carboxylic acid:
Figure imgf000141_0001
Step 1: Synthesis (lR,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8,lla-penta methyl-3a-((2-methyl-l -(pyrrolidin- 1 -yl)propan-2-yl)carbamoyl)-l -(prop-1 -en-2-yl) icosahydro-lH-cyclopenta[a ]chrysen-9-yl acetate:
Figure imgf000141_0002
To a stirred solution of 2-methyl-l -(pyrrolidin- l-yl)propan-2-amine (Intermediate 3, 2.2 g, 16.00 mmol, 2.0 eq) and triethylamine (6.0 mL, 40.0 mmol, 5.0 eq) in CH2C12 (20 ml) at O °C was added (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-(chlorocarbonyl)-5a, 5b, 8, 8,1 la-pentamethyl- l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (prepared as described in WO 2013/160810 A2, 4.0 g, 80.0 mmol, 1.0 eq) in CH2C12 (20 ml). The reaction mixture was allowed to stir at room temperature for overnight. The reaction mixture was diluted with water and extracted with CH2CI2. The combined organic extracts were dried over Na2S04, filtered and evaporated under reduced pressure. The residue was purified by column chromatography by using 3% methanol: DCM as an eluent to afford the desired product (2.5 g, yield: 51.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 6.79 (s, 1H), 4.65 (d, 2H), 4.38 (q, 1H), 3.55 (t, 4H), 3.44 (bs, 3H), 3.17 (d, 1H), 2.98 (q, 1H), 2.78-2.63 (m, 2H), 2.35-2.31 (m, 12H), 2.05 (bd, 1H), 1.99 (s, 3H), 1.95-1.87 (m, 3H), 1.61- 1.25 (m, 14H), 0.93 (s, 3H), 0.86 (s, 9H), 0.77 (s, 3H) and 0.65 (m, 3H); Mass: [M]+ 623.51 (100%).
Step 2: Synthesis of (lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-9-hydroxy-5a,5b,8,8, 11 a-pentamethyl-N -(2 -methyl- 1 -(pyrrolidin-l -yl)propan-2-yl)-l -(prop-1 -en-2-yl)icosahydro- 3aH-cyclopenta[a]chrysene-3a-carboxamide:
Figure imgf000142_0001
To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8, l la-pentamethyl-3a-((2-methyl-l-(pyrrolidin-l-yl)propan-2-yl)carbamoyl)-l-(prop-l-en-2- yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (step 1, 2.2 g, 40.0 mmol, 1.0 eq) in THF (18 ml) and Methanol (18 ml) was added potassium carbonate (4.8 g, 35.0 mmol, 10.0 eq). The reaction mixture was stirred at room temperature for about 48 hours. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was filtered through a pad of celite and washed with CH2C12. The filtrate was evaporated under reduced pressure and the crude was purified by column chromatography by using 3% methanol: dichloromethane as an eluent to afford the desired product (1.5 g, yield: 72.0%) as a white solid. 1H NMR (CDCI3, 300 MHz): δ 6.82 (s, 1H), 4.65 (d, 2H), 3.55 (t, 4H), 3.44 (bs, 4H), 3.17 (d, 1H), 2.98 (q, 1H), 2.78-2.63 (m, 2H), 2.35-2.31 (m, 12H), 2.05 (bd, 1H), 1.95-1.87 (m, 3H), 1.61-1.25 (m, 15H), 0.93 (s, 3H), 0.86 (s, 9H), 0.77 (s, 3H) and 0.65 (m, 3H); Mass: [M]+ 581.53 (100%).
Step 3: Synthesis of 1 -benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,llaR,llbR,13aR,13bR)-5a,5b,8,8, 1 la-pentamethyl- 3 a-((2 -methyl- 1 -(pyrrolidin-l -yl)propan-2-yl)carbamoyl)-l -(prop-1 -en-2- yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-dimethylcyclobutane-l,3- dicarboxylate:
Figure imgf000143_0001
To a stirred solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy- 5a,5b,8,8,l la-pentamethyl-N-(2-methyl-l-(pyrrolidin-l-yl)propan-2-yl)-l-(prop-l-en-2-yl) icosahydro-3aH-cyclopenta[a]chrysene-3a-carboxamide (step 2, 1.5 g, 2.5 mmol, 1.0 eq) in DCM (30 ml) then added (lS,3R)-3-((benzyloxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid (prepared as described in WO 2011/007230 A2, 1.0 g, 3.8 mmol, 1.5 eq) and the reaction mixture was cooled to 0 °C then added DCC (1.0 g, 5.0 mmol, 2.0 eq) followed by DMAP (0.063 g, 0.5 mmol, 0.2 eq). The reaction mixture was raised to room temperature and stirred for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was evaporated under reduced pressure and diluted with water. The aqueous layer was extracted with CH2CI2. The organic layer was washed with water and brine solution. The combined organic layers were dried over Na2S04, filtered and evaporated under reduced pressure. The crude residue was purified by column chromatography by using 4% MeOH and DCM as an eluent to obtain the desired product (1.2 g, yield: 58.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 7.36 (s, 5H), 6.79 (s, 1H), 5.48 (q, 2H), 4.65 (d, 2H), 4.36 (t, 1H), 3.00 (t, 2H), 2.82 (t, 3H), 2.34 (t, 2H), 2.14 (s, 1H), 1.90-1.83 (m, 6H), 1.63 (s, 3H), 1.57-1.47 (m, 4H), 1.43-1.28 (m, 22H), 1.18-0.98 (m, 4H) and 0.92-0.80 (m, 22H); Mass: [M]+ 825.64 (100%).
Step 4: synthesis of (lR,3S)-3-((((lS,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-l- isopropyl-5a,5b,8,8,lla-pentamethyl-3a-((2-methyl-l-(pyrrolidin-l-yl)propan-2-yl) carbamoyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclo butane-1 -carboxylic acid:
To a stirred solution of 1-benzyl 3-((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR) -5a,5b,8,8,l la-pentamethyl-3a-((2-methyl-l-(pyrrolidin-l-yl)propan-2-yl)carbamoyl)-l-(prop - 1 -en-2-yl)icosahydro- 1 H-cyclopenta[a] chry sen-9-yl) ( 1 R,3 S )-2,2-dimethylcyclobutane- 1,3- dicarboxylate (step 3, 0.5 g, 0.60 mmol) in ethyl acetate: methanol (5: 5 ml), was added 10% Pd/C (0.03 g) and purged with nitrogen. The reaction mixture was stirred for about 12 hours under H2 atmosphere. After completion of the reaction (monitored by TLC), the reaction mixture was filtered, concentrated and purified by silica gel column (elution 5% MeOH in
DCM) to afford the desired compound (130 mg, yield: 29%) as a white solid. 1H NMR (300 MHz, DMSO-d6): δ 6.80 (s, 1H), 4.37-4.31 (m, 1H), 2.82-2.79 (m, 2H), 2.62-2.59 (m, 6H), 2.35-2.31 (m, 3H), 2.16-2.12 (m, 2H), 1.92-1.82 (m, 2H), 1.72-1.03 (m, 35H), 0.91-0.88 (m,
9H), 0.85-0.79 (m, 12H), 0.72 (d, J= 6.6 Hz, 3H); ES Mass: 737.56 [M+H]+.
The below examples 97-102 were prepared by the procedure similar to example-96 by using corresponding intermediates inpresence of suitable reagents, reactents and solvents at appropriate reaction conditions. The corresponding intermediates and the charctrization data are mentioned in the below table.
Figure imgf000144_0001
Figure imgf000145_0001
Example 103: Preparation of qR,3Sy3-((((lS,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)- l-isopropyl-5a,5b,8,8 J la-pentamethyl-3a-((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidine-l- carbonyl)icosahvdro-lH-cvclopentaralchrvsen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane- 1-carboxylic acid:
Figure imgf000146_0001
Step 1: Synthesis of (lS,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-9-acetoxy-l-isopropyl- 5a,5b,8,8,lla-pentamethylicosahydro-3aH-cyclopenta[a]chrysene-3a-carboxylic acid:
Figure imgf000146_0002
To a solution of (lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-acetoxy-
5a,5b,8,8,l la-pentamethyl-l-(prop-l-en-2-yl)icosahydro-3aH-cyclopenta[a]chrysene-3a- carboxylic acid (prepared as described in WO 2013/160810 A2, 1.00 g, 0.82 mmol, 1.0 eq) in ethylacetate (40 mL) and methanol (10 mL) was added Pd/C (0.080 g, 0.49 mmol, 0.6 eq). The reaction mixture was stirred in hydrogen atmosphere at room temperature for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was filtered through a pad of celite and was washed with MeOH and DCM. The filtrate was evaporated under reduced pressure, the crude residue was purified by column chromatography by using 8% methanol and DCM as an eluent gave the desired compound (0.900 g, yield: 90.0%) as an off white solid.
Step 2: Synthesis of (lS,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-l-isopropyl-5a,5b,8,8, lla-pentamethyl-3a-((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidine-l-carbonyl)icosahydro-lH- cyclopenta [a]chrysen-9-yl acetate:
Figure imgf000146_0003
To a stirred solution of (S)-l-(pyrrolidin-2-ylmethyl)pyrrolidine (intermediate 39, 1.3 mL, 9.0 mmol, 1.5 eq) and triethylamine (2.4 mL, 18.0 mmol, 3.0 eq) in CH2C12 (60 ml) at 0 °C was added (lS,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-(chlorocarbonyl)-l- isopropyl-5a,5b,8,8,l la-pentamethylicosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (step 1, 3.0 g, 6.0 mmol, 1.0 eq) in CH2C12 (10 ml). The reaction mixture was allowed to stir at room temperature for overnight. The reaction mixture was diluted with water and extracted with CH2C12. The combined organic extracts were dried over Na2S04, filtered and evaporated under reduced pressure. The residue was purified by column chromatography by using 4% methanol: DCM as an eluent gave the desired product (1.8 g, yield: 78.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 4.40 (m, 1H), 4.14 (m, 1H), 3.48 (m, 2H), 2.93 (m, 1H), 2.30 (m, 1H), 2.08-2.05 (m, 1H), 1.99 (s, 3H), 1.82-1.52 (m, 14H), 1.37-1.16 (m, 15H), 1.08- 1.04 (m, 4H), 0.91-0.79 (m, 22H) and 0.60 (m, 3H); Mass: [M+H]+ 637.55 (100%).
Step 3: Synthesis of ((lS,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-9-hydroxy-l- isopropylSa, 5b, 8, 8, 1 la-pentamethylicosahydro -3aH-cyclopenta[a ]chrysen-3a-yl )((S)-2- (pyrrolidin-l -ylmethyl ) pyrrolidin-l -yl )methanone:
Figure imgf000147_0001
To a stirred solution of (lS,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-l-isopropyl -5a,5b,8,8,l la-pentamethyl-3a-((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidine-l-carbonyl) icosahydro-lH-cyclopenta[a]chrysen-9-yl acetate (step 2, 1.8 g, 2.83 mmol, 1.0 eq) in THF (15 ml), Methanol (15 ml) and water (20 mL) was added lithium hydroxide (0.63 g, 14.15 mmol, 5.0 eq). The reaction mixture was stirred at room temperature for about 12 hours. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was diluted with water and extracted in DCM. The organic layer was dride with Na2S04, and filtrate was evaporated under reduced pressure and the crude was purified by column chromatography by using 5% methanol: dichloromethane as an eluent gave the desired product (1.100 g, yield: 68.0%) as a white solid. 1H NMR (CDC13, 300 MHz): δ 4.26 (m, 1H), 4.12 (m, 1H), 3.47 (m, 2H), 2.96 (m, 2H), 2.30 (m, 1H), 2.08-2.05 (m, 1H), 1.82- 1.52 (m, 14H), 1.37-1.16 (m, 15H), 1.08-1.04 (m, 4H), 0.91-0.79 (m, 22H) and 0.60 (m, 3H); Mass: [M+H]+ 595.58 (100%).
Step 4: Synthesis of 1 -benzyl 3-((lS,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-l-isopropyl -5a,5b,8,8,lla-pentamethyl-3a-((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidine-l-carbonyl) icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-dimethylcyclobutane-l,3- dicarboxylate:
Figure imgf000147_0002
To a stirred solution of ((lS,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-9-hydroxy- l-isopropyl-5a,5b,8,8,l la-pentamethylicosahydro-3aH-cyclopenta[a]chrysen-3a-yl)((S)-2- (pyrrolidin-l-ylmethyl)pyrrolidin-l-yl)methanone (step 3, 1.100 g, 1.85 mmol, 1.0 eq) in
DCM (50 ml) then added (lS,3R)-3-((benzyloxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid (prepared as described in WO 2011/007230 A2, 0.73 g, 2.77 mmol, 1.5 eq) this reaction mixture was cooled to 0 °C then added DCC (0.76 g, 3.7 mmol, 2.0 eq) followed by DMAP (0.04 g, 0.37 mmol, 0.2 eq). The reaction mixture was raised to room temperature stirred for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was evaporated under reduced pressure and diluted with water. The aqueous layer was extracted with CH2CI2. The organic year was washed with water and brine solution. The combined organic layers were dried over Na2S04, filtered and evaporated under reduced pressure. The crude residue was purified by column chromatography by using 5% MeOH and DCM as an eluent to obtain the desired product (0.700 g, yield: 46.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 7.36 (m, 5H), 5.14 (q, 2H), 4.36 (m, 1H), 4.14 (bs, 1H), 3.48 (m, 2H), 2.96-2.80 (m, 4H), 2.42-2.30 (m, 4H), 2.06-2.56 (m, 19H), 1.41-1.19 (m, 15H), 1.10-1.05 (m, 4H), 0.95-0.90 (m, 6H), 0.86- 0.80 (m, 16H) and 0.74 (m, 3H); Mass: [M+H]+ 839.68 (60%).
Step 4: Synthesis of (lR,3S)-3-((((lS,3aS,5aR,5bR, 7aR,9S,llaR,llbR,13aR,13bR)-l- isopropyl-5a,5b,8,8 la^entamethyl-3a-((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidine-l- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane- 1 -carboxylic acid:
To a solution of 1-benzyl 3-((lS,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-l- isopropyl-5a,5b,8,8,l la-pentamethyl-3a-((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidine-l- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) (lR,3S)-2,2-dimethylcyclobutane-l,3- dicarboxylate (step 3, 0.700 g, 0.83 mmol, 1.0 eq) in methanol (10 mL) and ethylacetate (30 mL) then added Pd/C (0.02 g, 0.23 mmol, 0.28 eq). The reaction mixture was in H2 gas atmosphere and stirred at room temperature for overnight. TLC indicated starting material was consumed and the desired product was observed. The reaction mixture was filtered through a pad of celite and was washed with MeOH and DCM. The filtrate was evaporated under reduced pressure, the crude residue was purified by column chromatography by using 10% methanol: DCM as an eluent gave the desired compound (0.080g, yield: 13.0%) as a white solid. 1H NMR (DMSO-d6, 300 MHz): δ 12.13 (s, 1H), 4.37 (m, 1H), 4.17 (bs, 1H), 3.49 (m, 3H), 3.02 (m, 3H), 2.87-2.72 (m, 4H), 2.34-2.23 (m, 3H), 2.07-1.85 (m, 11H), 1.69- 1.56 (m, 5H), 1.41-1.19 (m, 15H), 1.10-1.05 (m, 4H), 0.95-0.90 (m, 6H), 0.86-0.80 (m, 16H) and 0.74 (m, 3H); Mass: [M+H]+ 749.66 (100%); HPLC: 96.84%.
PHARMACOLOGICAL ACTIVITY The compounds described herein can be tested for their biological activity following procedures known to a person of ordinary skill in the art. For example, the following protocols can be employed for testing the compounds. These protocols are illustrative and do not limit to the scope of the invention.
Example 104: Evaluation of compounds antiviral activity:
MT2 cells were infected with HIV-1 strain 92HT599 (10 TCID 50/ 30000 cells). The infected cells were plated at the concentration of -30,000 cells per well in 96 well plate. Test compound was added to the micro plate in defined format with the final concentration of DMSO (vehicle) is not more than 1%. Incubation was carried out in C02 incubator for - 96 hours for viral infection. At the end of incubation period an aliquot from each well was taken for p24 estimation. The quantitation of p24 is an index for antiviral activity of the compound. Percent inhibition was calculated with reference to control values (vehicle controls).
P-24 estimation was carried out using Advance biosciences kit as per the procedure detailed by supplier.
For 0% serum binding assay, wherein "A" refers to an IC50 value < 10 nM, "B" refers to IC50 value in range of 10.01-50 nM, and "C" refers to IC50 value > 50 nM.
For 45 mg/ml HSA serum binding assay, wherein "A" refers to an IC50 value < 50 nM, "B" refers to IC50 value in range of 50.01-100 nM, and "C" refers to IC50 values > 100 nM. The IC50 (nM) values are set forth in Table- 1.
TABLE- 1
Figure imgf000150_0001
References:
1. Antiviral methods and protocols (Eds: D Kinchington and R F Schinazi) Humana Press Inc., 2000.
2. HIV protocols (Eds: N L Michael and J H Kim) Humana Press Inc, 1999.
3. DAIDS Virology manual from HIV laboratories, Publication NIH-97-3838, 1997.
4. HIV-1 p24 antigen capture assay, enzyme immunoassay for detection of Human immunodeficiency Virus Type 1 (HIV-1) p24 in tissue culture media - Advanced bio science laboratories, Inc kit procedure.
Although the invention herein has been described with reference to particular embodiments, it is to be understood that these embodiments are merely illustrative of the principles and applications of the present invention. It is therefore to be understood that numerous modifications may be made to the illustrative embodiments and that other arrangements may be devised without departing from the spirit and scope of the present invention as described above.
All publications and patent applications cited in this application are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated herein by reference.

Claims

A com ound of the formula (I)
Figure imgf000152_0001
wherein,
Figure imgf000152_0002
(wherein R is hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl);
R2 is hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted aminoacids, substituted or unsubstituted alkoxy or substituted or unsubstituted cycloalkyl;
X is absent, O, S, CH2 or NRa (wherein Ra can be H, C(0)Rc, C(S)RC, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, or Ra is with their adjacent N and Carbon together form N-contained heterocyclyl ring; Y is C(O), C(S) or CReRf (wherein Re and Rf are independently selected from H, substituted or unsubstituted alkyl);
R3 and R4 are 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 R3 and R4 are taken together with the carbon atoms to which they are attached to form a bond, or R3 and R4 are taken together with the carbon atoms to which they are attached to form cycloalkyl, or R3 and R4 are taken together with the carbon atoms to which they are attached to form epoxide;
R5 and Rc are independently selected from H, C02Rd (wherein Rd is H, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl), or substituted or unsubstituted alkyl;
R6 and R8 are independently selected from H, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl;
R7 is substituted or unsubstituted alkyl or substituted or unsubstituted cycloalkyl;
R6 and R7 are taken together with their nitrogen and carbon to which they are attached to form a substituted or unsubstituted 4-7 membered heterocyclyl, substituted or unsubstituted 4-7 membered bridged heterocyclyl wherein the substituents are alkyl, heterocyclyl or -O-heterocyclyl;
R7 and R8 are taken together with the carbon atom to which they are attached to form 3- 7 membered cycloalkyl or heterocyclyl;
/ ;
''·—■'' is 4-10 membered heterocyclyl or 4-10 membered heteroaryl;
Zi, Z2 and Z3 are independently selected from H, hydroxy, halo, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkylsulfonyl, substituted or unsubstituted alkoxy, substituted or unsubstituted alkoxylalkoxy substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, substituted or unsubstituted heterocyclylalkyl, substituted or unsubstituted aminoalkyl, substituted or unsubstituted hydroxyalkyl, -O-heterocyclyl, substituted or unsubstituted heteroaryl; wherein the substituents are aryl, alkyl, alkoxy or alkoxylalkoxy including pharmaceutically acceptable salts, pharmaceutically acceptable solvates, pharmaceutically acceptable hydrates, tautomers, stereoisomers, ester prodrugs, or combination thereof.
2. The compound according to claim 1, is a compound of the formula (IA):
Figure imgf000154_0001
wherein,
Ri, R2, R3, R4, R5, R7, R8, 'X', Re, Rf, Zi, Z2 and Z3 including pharmaceutically acceptable salts, pharmaceutically acceptable solvates, pharmaceutically acceptable hydrates, tautomers, stereoisomers, ester prodrugs, or combination thereof.
3. The compound la (IB):
Figure imgf000154_0002
wherein
Ri, R2, R3, R4, R5, R8, 'X', Ύ', Zi, Z2 and Z3 including pharmaceutically acceptable salts, pharmaceutically acceptable solvates, pharmaceutically acceptable hydrates, tautomers, stereoisomers, ester prodrugs, or combination thereof.
4. The compound according to claim 1-3, wherein Ri is o ,
Figure imgf000154_0003
or
Figure imgf000154_0004
5. The compound according to claim 1 and 2, wherein R6 is hydrogen.
6. The compound according to claim 1 and 2, wherein R7 is substituted or
unsubstituted alkyl and Rg is hydrogen or substituted or unsubstituted alkyl.
7. The compound according to claim 1 to 6, wherein ring
Figure imgf000154_0005
' is selected from morpholinyl, pyrrolidinyl, piperidinyl, piperizinyl, azepane and 8- azabicyclo[3.2.1]octane, pyrrole, imidazoylyl, pyrazolyl, lH-pyrrolo[2,3- b]pyridine, thiomorpholine 1,1 -dioxide and thiomorpholinyl.
8. Acompound is selected from the group consisting of:
2,2-dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8, 11 a-pentamethyl-3 a-((2-methyl- 1 -morpholinopropan-2-yl)carbamoyl) - 1 -( 1 -methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)butanoic acid,
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-((2-methyl-l-(piperidin-l-yl)propan-2-yl)carbamoyl)-l-(l-methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)butanoic acid,
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-((2-methyl-l-(pyrrolidin-l-yl)propan-2-yl)carbamoyl)-l-(l-methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)butanoic acid,
4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-((lR,5S)-3-(3-isopropyl- 5-methyl-4H-l,2,4-triazol-4-yl)-8-azabicyclo[3.2.1]octan-8-yl)-2-methylpropan-2-yl) carbamoyl)-5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a] chrysen-9-yl)oxy)-2,2-dimethyl-4-oxobutanoic acid,
2,2-dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8, 1 la-pentamethyl- l-(l-methylcyclopropyl)-3a-((S)-2-(pyrrolidin- l-ylmethyl)pyrrolidine- 1- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)butanoic acid,
2,2-dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8,l la- pentamethyl- 1 - ( 1 -methylcyclopropyl) - 3a-((S)-2- (pyrrolidine- 1 -c arbonyl)pyrrolidine- 1 - carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)butanoic acid,
4- (((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-ethylpiperazin-l- yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl- 1-(1 -methylcyclopropyl) icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)-2,2-dimethyl-4-oxobutanoic acid,
5- (((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-ethylpiperazin-l- yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl- 1-(1 -methylcyclopropyl) icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)-3,3-dimethyl-5-oxopentanoic acid,
5-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-ethylpiperidin-l- yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl- 1-(1 -methylcyclopropyl) icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)-3,3-dimethyl-5-oxopentanoic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-((R)-3- hydroxypyrrolidin- l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b, 8, 8,1 la-pentamethyl- 1-(1- methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl cyclobutane-l-carboxylic acid, (lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8, 11 a-pentamethyl-3a-((2-methyl- 1 -morpholinopropan-2-yl)carbamoyl)- 1 -( 1 -methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid,
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8, 11 a-pentamethyl-3a-((2-methyl- 1 -(pyrrolidin- 1 -yl)propan-2-yl)carbamoyl)- 1 -( 1 -methyl cyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l- carboxylic acid,
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8, 11 a-pentamethyl-3a-((2-methyl- 1 -(piperidin- 1 -yl)propan-2-yl)carbamoyl)- 1 -( 1 -methyl cyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l- carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-((lR,5S)-3-(3- isopropyl-5-methyl-4H-l,2,4-triazol-4-yl)-8-azabicyclo[3.2.1]octan-8-yl)-2-methylpropan-2- yl)carbamoyl)-5a,5b,8,8, 1 la-pentamethyl- l-(l-methylcyclopropyl)icosahydro- lH-cyclopenta [a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l-carboxylic acid,
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,1 la-pentamethyl- 1-(1 -methylcyclopropyl)-3a-((S)-2-(pyrrolidin- l-ylmethyl)pyrrolidine- l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l- carboxylic acid,
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,l la-pentamethyl-3a-((2-methyl-l-((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidin-l-yl)propan-2- yl)carbamoyl)-l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy) carbonyl)cyclobutane-l-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(4-ethyl piperazin-l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b,8,8,l la-pentamethyl- l-(l-methyl cyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl cyclobutane-l-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-ethyl piperazin- 1 -yl)methyl)pyrrolidine- 1 -carbonyl)-5a,5b, 8,8, 11 a-pentamethyl- 1 -( 1 -methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-ethyl piperidin- l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl- 1-(1 -methylcyclo propyl)icosa ydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid,
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,l la-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-(piperidin-l-ylmethyl)pyrrolidine-l carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid,
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,l la-pentamethyl-l-(l-methylcyclopropyl)-3a-((R)-2-(pyrrolidin-l-ylmethyl)pyrrolidine- l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l- carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(azepan-l- ylmethyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclopropyl) icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((lH- imidazol- l-yl)methyl)pyrrolidine- l-carbonyl)-5a,5b,8,8, 11 a-pentamethyl- l-(l-methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((2- isopropyl-lH-imidazol-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl-l-(l methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl cyclobutane-1 -carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(((S)-3- (dimethylamino)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl- l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2- dimethylcyclobutane-1 -carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((3,5- dimethyl-lH-pyrazol-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8, 11 a-pentamethyl- 1-(1- methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl cyclobutane-1 -carboxylic acid,
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8, 11 a-pentamethyl- l-(l-methylcyclopropyl)-3a-((S)-2-(pyrrolidine- l-carbonyl)pyrrolidine- 1- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid, (lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,1 la-pentamethyl- 1-(1 -methylcyclopropyl)-3a-((S)-2-(morpholinomethyl pyrrolidine- 1- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4- (hydroxymethyl)piperidin-l-yl)methyl)pyrroM
(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl cyclobutane-l-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-butyl piperazin- 1 -yl)methyl)pyrrolidine- 1 -carbonyl)-5a,5b,8,8, 11 a-pentamethyl- 1 -( 1 -methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid,
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,1 la-pentamethyl- l-(l-methylcyclopropyl)-3a-((S)-2-((4-methylpiperidin-l-yl)methyl) pyrrolidine- l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane- 1 -carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4- isopropylpiperazin- 1 -yl)methyl)pyrrolidine- 1 -carbonyl)-5a,5b, 8,8, 11 a-pentamethyl- 1 -( 1 - methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl cyclobutane-l-carboxylic acid,
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,1 la-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-(((S)-2-(5-phenyl-lH-imidazol-2- yl)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) oxy)carbonyl)cyclobutane- 1 -carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((lH- pyrrolo[2,3-b]pyridin- l-yl)methyl)pyrrolidine- l-carbonyl)-5a,5b, 8, 8,1 la-pentamethyl- 1-(1- methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl cyclobutane-l-carboxylic acid,
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,1 la-pentamethyl- l-(l-methylcyclopropyl)-3a-((S)-2-(((R)-2-methylpyrrolidin-l-yl) methyl)pyrrolidine-l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl) cyclobutane-l-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4,4- difluoropiperidin- 1 -yl)methyl)pyrrolidine- 1 -carbonyl)-5a,5b, 8,8, 11 a-pentamethyl- 1 -( 1 - methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl cyclobutane-l-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((3,3- difluoropyrrolidin- 1 - yl)methyl)pyrrolidine- 1 -c arbonyl) -5 a, 5b , 8 , 8 , 11 a-pentamethyl- 1 - ( 1 - methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl cyclobutane-l-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-fluoro piperidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(((S)-3- fluoropyrrolidin- 1 - yl)methyl)pyrrolidine- 1 -c arbonyl) -5 a, 5b , 8 , 8 , 11 a-pentamethyl- 1 - ( 1 -methyl cyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclo butane- 1-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(((S)-2-(2- hydroxypropan-2-yl)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-penta methyl- l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)- 2,2-dimethylcyclobutane- 1-carboxylic acid,
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,1 la-pentamethyl-l-(l-methylcyclopropyl)-3a-((2S,4R)-2-(pyrrolidin-l-ylmethyl)-4-(((R)- tetrahydrofuran-3-yl)oxy)pyrrolidine-l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) oxy)carbonyl)cyclobutane- 1-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(((S)-2- (hydroxymethyl)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl- l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2- dimethylcyclobutane- 1-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(((S)-3-(2- methoxyethoxy)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl-l- (l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl cyclobutane-l-carboxylic acid,
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,1 la-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-(((R)-3-(((R)-tetrahydrofuran-3-yl) oxy)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9- yl)oxy)carbonyl)cyclobutane- 1-carboxylic acid, (lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,1 la-pentamethyl-l-(l-methylcyclopropy
bipyrrolidine]- -carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl) cyclobutane-l-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-cyclo hexylpiperazin- 1 - yl)methyl)pyrrolidine- 1 -c arbonyl) - 5a,5b,8,8,l l a-pentamethyl- 1 - ( 1 -methyl cyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclo butane- 1-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-(2- methoxyethyl)piperazin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b, 8,8,11 a-pentamethyl- 1 - (l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl cyclobutane-l-carboxylic acid,
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,l la-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-((4-methylpiperazin-l-yl)methyl) pyrrolidine- l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane- 1-carboxylic acid,
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,11 a-pentamethyl- 1 - ( 1 -methylcyclopropyl) - 3a-((S)-2- (thiomorpholinomethyl)pyrrolidine- l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l- carboxylic acid,
(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-((2-methyl-lH-imidazol-l-yl)methyl)pyrrolidine-l-carbonyl)- l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl) cyclobutane-l-carboxylic acid,
(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-lH-imidazol-l-yl)methyl)pyrrolidine-l-carbonyl)- l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl) cyclobutane-l-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((lR,2S,5S)-6,6- dimethyl-2-(pyrrolidin-l-ylmethyl)-3-azabicyclo[3.1.0]hexane-3-carbonyl)-5a,5b, 8,8,1 la- pentamethyl-l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane- 1-carboxylic acid,
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,l la-pentamethyl-3a-((2-methyl-l-(4-(methylsulfonyl)piperazin-l-yl)propan-2-yl) carbamoyl)- l-(l-methylcyclopropyl)icosa ydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl) cyclobutane-l-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(((2S,4S)-4- iluoro-2-((2-methoxyethoxy)methyl)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b, 8,8,l la-pentamethyl-l-(l-methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl) oxy)carbonyl)-2,2-dimethylcyclobutane- 1 -carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((l,l- dioxidothiomorpholino)methyl)pyrrolidine-l-carbonyl)-5a,5b, 8,8,11 a-pentamethyl- 1-(1- methylcyclopropyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl cyclobutane-l-carboxylic acid,
(lS,3R)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,l la-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidine- l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l- carboxylic acid,
(lS,3R)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-ethyl piperazin- 1 -yl)methyl)pyrrolidine- 1 -carbonyl)-5a,5b,8,8, 11 a-pentamethyl- 1 -( 1 -methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid,
(lS,3R)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-fluoro piperidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl-l-(l-methylcyclo propyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid,
(lS,3R)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,l la-pentamethyl-l-(l-methylcyclopropyl)-3a-((S)-2-(morpholinomethyl)pyrrolidine-l- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid,
(lS,3R)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,l la-pentamethyl-l-(l-methylcyclopropyl)-3a-((R)-2-(pyrrolidin-l-ylmethyl)pyrrolidine- l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l- carboxylic acid,
2,2-dimethyl-4-oxo-4-((( lR,3aS,5aR,5bR,7aR,9S, 11 aR, 1 IbR, 13aR, 13bR)-5a,5b,8,8, l la-pentamethyl-3a-((2-methyl-l-(piperidin-l-yl)propan-2-yl)carbamoyl)-l-(prop-l-en-2-yl) icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)butanoic acid, 4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(4-ethylpiperazin-l-yl)-2 methylpropan-2-yl)carbamoyl)-5a,5b,8,8,l la-pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH cyclopenta[a]chrysen-9-yl)oxy)-2,2-dimethyl-4-oxobutanoic acid,
2,2-dimethyl-4-oxo-4-(((lR,3aS,5aR,5bR,9S,l laR,l lbR,13aR,13bR)-5a,5b,8,8,l la- pentamethyl-l-(prop-l-en-2-yl)-3a-((S)-2-(pyrrolidine-l-carbonyl)pyrrolidine-l-carbonyl) icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)butanoic acid,
4-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(((R)-3-(2-methoxy ethoxy)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl-l-(prop-l en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)-2,2-dimethyl-4-oxobutanoic acid,
4- (((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-ethylpiperazin-l- yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl-l-(prop-l-en-2-yl)icosahydro- lH-cyclopenta[a]chrysen-9-yl)oxy)-2,2-dimethyl-4-oxobutanoic acid,
5- (((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(4-ethylpiperazin-l-yl)-2 methylpropan-2-yl)carbamoyl)-5a,5b,8,8,l la-pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH cyclopenta[a]chrysen-9-yl)oxy)-3,3-dimethyl-5-oxopentanoic acid,
5-(((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-ethylpiperazin-l- yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl-l-(prop-l-en-2-yl)icosahydro- lH-cyclopenta[a]chrysen-9-yl)oxy)-3,3-dimethyl-5-oxopentanoic acid,
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8, 1 la-pentamethyl-3a-((2-methyl-l-(pyrrolidin- l-yl)propan-2-yl)carbamoyl)- l-(prop- 1-en- 2-yl)icosahydro- lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane- 1-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(4-ethyl piperazin-l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b,8,8,l la-pentamethyl-l-(prop-l-en-2- yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid,
(lR,3R)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8, l la-pentamethyl-3a-((2-methyl-l-(piperidin-l-yl)propan-2-yl)carbamoyl)-l-(prop- 1-en- 2-yl)icosahydro- lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane- 1-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(4-benzyl piperazin-l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b,8,8,l la-pentamethyl-l-(prop-l-en-2- yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid,
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8, 1 la-pentamethyl- l-(prop- l-en-2-yl)-3a-((S)-2-(pyrrolidine- l-carbonyl)pyrrolidine- 1- carbonyl)icosa ydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-((R)-3-(2- methoxyethoxy)pyrrolidin- l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b,8,8, 1 la-pentamethyl -l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl cyclobutane-l-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(4-(2-methoxy ethoxy)piperidin-l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b,8,8,l la-pentamethyl- l-(prop- l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane- 1-carboxylic acid,
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,11 a-pentamethyl- 1 -(prop- 1 -en-2-yl)-3a-(( 1 -(pyrrolidin- 1 -ylmethyl)cyclobutyl) carbamoyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid,
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8,11 a-pentamethyl- 1 -(prop- 1 -en-2-yl)-3a-(( 1 -(pyrrolidin- 1 -ylmethyl)cyclopentyl) carbamoyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((l-(4-ethoxy-4- phenylpiperidin- l-yl)-2-methylpropan-2-yl)carbamo yl)-5a,5b, 8, 8,1 la-pentamethyl- l-(prop- l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane- 1-carboxylic acid,
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8, 1 la-pentamethyl- l-(prop- l-en-2-yl)-3a-(((S)- 1 -(pyrrolidin- l-yl)propan-2-yl)carbamoyl) icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid,
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8, 1 la-pentamethyl-3a-(((S)-4-methyl- l-(piperidin- l-yl)pentan-2-yl)carbamoyl)-l-(prop- 1- en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-ethyl piperazin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b, 8, 8,1 la-pentamethyl- l-(prop-l -en-2- yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid, (lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8, 1 la-pentamethyl-3a-((S)-2-(piperidin- l-ylmethyl)pyrrolidine-l-carbonyl)- l-(prop- l-en-2 yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((4-ethyl piperidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl-l-(prop-l-en-2-yl) icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((3,5- dimethyl-lH-pyrazol-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl-l- (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclo butane- 1-carboxylic acid,
(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-(morpholinomethyl)pyrrolidine-l-carbonyl)-l-(prop-l-en-2- yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane- 1-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((lH- pyrazol-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl-l-(prop-l-en-2-yl) icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(((R)-3-(2- methoxyethoxy)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethyl^ (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclo butane- 1-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(((R)-3-(2- methoxyethoxy)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-pentamethy (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclo butane- 1-carboxylic acid,
(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-((R)-2-(pyrrolidin-l-ylmethyl)pyrrolidine-l- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane- 1-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(((S)-2-(2- hydroxypropan-2-yl)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)-5a,5b,8,8,l la-penta methyl- l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2- dimethylcyclobutane- 1-carboxylic acid, (lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((lR,2S,5S)-6,6- dimethyl-2-(pyrrolidin-l-ylmethyl)-3-azabicyclo[3.1.0]hexane-3-carbonyl)-5a,5b, 8,8,1 la- pentamethyl-l-(prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2- dimethylcyclobutane-l-carboxylic acid,
(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-((2-(pyrrolidin-l-ylmethyl)-lH-pyrrol-l- yl)methyl)pyrrolidine-l-carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl) cyclobutane-l-carboxylic acid,
(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-((2-methyl-lH-pyrrol-l-yl)methyl)pyrrolidine-l-carbonyl)-l- (prop-l-en-2-yl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l- carboxylic acid,
(lR,3S)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8, 1 la-pentamethyl- l-(prop- l-en-2-yl)-3a-((S)-2-(pyrrolidin- l-ylmethyl)pyrrolidine- 1- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid,
(lR,3S)-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-((lH-pyrrol- l-yl)methyl)pyrrolidine- l-carbonyl)-5a,5b,8,8, 1 la-pentamethyl- 1 -(prop- l-en-2-yl) icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid,
(lS,3R)-2,2-dimethyl-3-((((lR,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-5a,5b, 8,8, 1 la-pentamethyl- l-(prop- l-en-2-yl)-3a-((S)-2-(pyrrolidin- l-ylmethyl)pyrrolidine- 1- carbonyl)icosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)cyclobutane-l-carboxylic acid,
(lR,3S)-3-((((lS,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-l-isopropyl-5a,5b, 8,8,l la-pentamethyl-3a-((2-methyl-l-(pyrrolidin-l-yl)propan-2-yl)carbamoyl)icosahydro- lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l-carboxylic acid,
(lR,3S)-3-((((lS,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-l-isopropyl-5a,5b,8,8, l la-pentamethyl-3a-((l-(pyrrolidin-l-ylmethyl)cyclobutyl)carbamoyl)icosahydro-lH-cyclo penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l-carboxylic acid,
(lR,3S)-3-((((lS,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-l-isopropyl-3a-((l- ((R)-3-(2-methoxyethoxy)pyrrolidin- l-yl)-2-methylpropan-2-yl)carbamoyl)-5a,5b,8,8, 1 la- pentamethylicosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclo butane- 1 -carboxylic acid, (lR,3S)-3-((((lS,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-l-isopropyl-3a-((l-(4- (2-methoxyethoxy)piperidin- 1 -yl)-2-methylpropan-2-yl)carbamoyl)-5 a,5b, 8 ,8,11 a-penta methylicosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l- carboxylic acid,
(lR,3S)-3-((((lS,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-l-isopropyl-5a,5b,8,8, l la-pentamethyl-3a-((l-(pyrrolidin-l-ylmethyl)cyclopentyl)carbamoyl)icosa ydro-lH-cyclo penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l-carboxylic acid,
(lR,3S)-3-((((lS,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-l-isopropyl-5a,5b,8,8, l la-pentamethyl-3a-(((S)-4-methyl-l-(pyrrolidin-l-yl)pentan-2-yl)carbamoyl)icosahydro- lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l-carboxylic acid, (lR,3S)-3-((((lS,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-3a-((S)-2-(((S)-2-(2- hydroxypropan-2-yl)pyrrolidin-l-yl)methyl)pyrrolidine-l-carbonyl)-l-isopropyl-5a,5b,8,8, l la-pentamethylicosahydro-lH-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclo butane- 1 -carboxylic acid; and
(lR,3S)-3-((((lS,3aS,5aR,5bR,7aR,9S,l laR,l lbR,13aR,13bR)-l-isopropyl-5a,5b,8,8, l la-pentamethyl-3a-((S)-2-(pyrrolidin-l-ylmethyl)pyrrolidine-l-carbonyl)icosa ydro-lH- cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-l-carboxylic acid including pharmaceutically acceptable salts, pharmaceutically acceptable solvates, pharmaceutically acceptable hydrates, tautomers, stereoisomers, ester prodrugs, or combination thereof.
9. A pharmaceutical composition comprising a compound according to any one of claims 1-8 and at least one pharmaceutically acceptable excipient.
10. The pharmaceutical composition according to claim 9, wherein the pharmaceutically acceptable excipient is a carrier or diluent.
11. A method for preventing, ameliorating or treating a viral mediated disease, disorder or syndrome in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1-8.
12. The method according to claim 11, wherein the viral mediated disease, disorder or syndrome is HIV infection, HBV, HCV, a retroviral infection genetically related to AIDS, respiratory disorders (including adult respiratory distress syndrome (ARDS)), inflammatory disease, or a combination thereof.
13. A method of treating HIV in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1-8.
14. A method for preventing, ameliorating or treating an HIV mediated disease in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition according to claim 9.
15. The method according to claim 14, wherein the HIV mediated disease is HIV infection, HBV infection, or HCV infection.
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US9868758B2 (en) 2014-06-30 2018-01-16 Hetero Labs Limited Betulinic proline imidazole derivatives as HIV inhibitors
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US10370405B2 (en) 2015-03-16 2019-08-06 Hetero Labs Limited C-3 novel triterpenone with C-28 amide derivatives as HIV inhibitors
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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
JP2020534358A (en) * 2017-09-18 2020-11-26 上海 インスティテュート オブ マテリア メディカ、チャイニーズ アカデミー オブ サイエンシーズＳｈａｎｇｈａｉ Ｉｎｓｔｉｔｕｔｅ Ｏｆ Ｍａｔｅｒｉａ Ｍｅｄｉｃａ， Ｃｈｉｎｅｓｅ Ａｃａｄｅｍｙ Ｏｆ Ｓｃｉｅｎｃｅｓ Pentacyclotriterpene compounds and methods for producing them, drug compositions and uses
US11192916B2 (en) 2017-09-18 2021-12-07 Shanghai Institute Of Materia Medica, Chinese Academy Of Sciences Pentacyclic triterpene compound and preparation method therefor, and pharmaceutical composition and use thereof
JP7029542B2 (en) 2017-09-18 2022-03-03 上海 インスティテュート オブ マテリア メディカ、チャイニーズ アカデミー オブ サイエンシーズ Pentacyclotriterpene compounds and methods for producing them, drug compositions and uses
CN107663184A (en) * 2017-11-15 2018-02-06 上海皓伯化工科技有限公司 A kind of synthetic method of the hydroxy piperidines of N Boc 4
EP3924361B1 (en) * 2019-02-11 2023-09-06 Hetero Labs Limited Novel triterpene derivatives as hiv inhibitors

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