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WO2011080562A1 - Novel aza-peptides containing 2,2-disubstituted cyclobutyl and/or substituted alkoxy benzyl derivatives as antivirals - Google Patents

Novel aza-peptides containing 2,2-disubstituted cyclobutyl and/or substituted alkoxy benzyl derivatives as antivirals Download PDF

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Publication number
WO2011080562A1
WO2011080562A1 PCT/IB2010/003313 IB2010003313W WO2011080562A1 WO 2011080562 A1 WO2011080562 A1 WO 2011080562A1 IB 2010003313 W IB2010003313 W IB 2010003313W WO 2011080562 A1 WO2011080562 A1 WO 2011080562A1
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methyl
hydroxy
substituted
unsubstituted
hydrazinyl
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French (fr)
Inventor
Bandi Parthasaradhi Reddy
Bandi Vamsi Krishna
Vedula Manohar Sharma
Kura Rathnakar Reddy
Musku Madhanmohan Reddy
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Hetero Research Foundation
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Hetero Research Foundation
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/24—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
    • C07D213/36—Radicals substituted by singly-bound nitrogen atoms
    • C07D213/42—Radicals substituted by singly-bound nitrogen atoms having hetero atoms attached to the substituent nitrogen atom
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/18—Antivirals for RNA viruses for HIV
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C271/00—Derivatives of carbamic acids, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups
    • C07C271/06—Esters of carbamic acids
    • C07C271/08—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms
    • C07C271/10—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms
    • C07C271/22—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms to carbon atoms of hydrocarbon radicals substituted by carboxyl groups
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
    • C07D215/02—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom
    • C07D215/04—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, directly attached to the ring carbon atoms
    • C07D215/06—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, directly attached to the ring carbon atoms having only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, attached to the ring nitrogen atom
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
    • C07D295/04—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms
    • C07D295/08—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly bound oxygen or sulfur atoms
    • C07D295/084—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly bound oxygen or sulfur atoms with the ring nitrogen atoms and the oxygen or sulfur atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings
    • C07D295/088—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly bound oxygen or sulfur atoms with the ring nitrogen atoms and the oxygen or sulfur atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings to an acyclic saturated chain

Definitions

  • the present invention relates to novel aza-peptides containing 2,2- disubstituted cyclobutyl and/or substituted alkoxy benzyl derivatives and compositions for inhibiting Human Immunodeficiency Virus (HIV) and process for making the compounds.
  • HIV Human Immunodeficiency Virus
  • HIV- 1 infection remains a major medical problem, with an estimated 34 million people infected worldwide.
  • the number of cases of HIV and AIDS (acquired immunodeficiency syndrome) has risen rapidly. In 2009. approximately 2.6 million new infections were reported, and 2.0 million people died from AIDS.
  • HIV protease inhibitors are one important class of therapeutic agents for inhibition and treatment of HIV infection.
  • protease inhibitors like saquinavir, ritonavir, indinavir, nelfinavir, amprenavir, lopinavir, fosamprenavir and atazanavir have been approved as drugs for treatment of HIV infection.
  • novel HIV inhibitors that are very potent and effective against resistant strains of HIV. Therefore, novel anti-HIV agents exhibiting distinct resistance patterns, and favorable pharmacokinetic as well as safety profiles are needed to provide more treatment options.
  • Aza-peptide structural classes of compounds as HIV- protease inhibitors were explored intensively for their therapeutic activities and those include for example, the patent publications: WO 97/40029, US0584991 1 and US06087383 disclosed antivirally active heterocyclic azahexane derivatives; WO 94/ 19332 disclosed retroviral protease inhibiting compounds; WO 97/ 19055 disclosed azahexane derivatives as substrate isosters of retroviral aspirate proteases; WO 98/03476 disclosed anilinopeptide derivatives; and WO 08/ 1 56632 disclosed azapeptide derivatives.
  • no patent or publication in the literature has reported the specific compounds or their anti HIV activity of the present invention. This observation is of practical importance in synthesizing the compounds of the present invention and screening them for their anti-HIV activity, for the discovery of new drugs.
  • Wj and W 2 are independently selected from a bond or substituted or unsubstituted alkylene and preferably the substituent is selected from n-butyl, isobutyl or tertiary butyl ;
  • Zi and Z 2 are independently selected from a bond or substituted or unsubstituted cyclo butyl ; provided that when atleast one of the Zi and Z 2 are substituted or unsubstituted cyclo butyl then Ri can be H, -0[C(R a )2] m -alkyl, - 0[C(R b )2] m -cycloalkyl, -0[C(R c )2] m -heterocyclyl, substituted or unsubstituted phenyl, substituted or unsubstituted pyridine, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl; or when both Zi and Z 2 are a bond then Ri can be -0[C(R a )2] n -Ri a > wherein Ri a can be H, substituted or unsubstituted alkyl, substituted or unsubstit
  • R 2 , R 3 , R 2a and R 3a are independently can be selected from H, C(0)R", C(0)OR", or S(0) : R";
  • R can be H, OH, halogen, NR", C(0)OR", C(0)NR" substituted or unsubstituted alkyl, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl;
  • n can be an integer 0-3;
  • n can be an integer 1 -3 ;
  • R a , R b and R c are independently can be selected from H, or substituted or unsubstituted alkyl ;
  • R" can be H, substituted or unsubstituted alky], substituted or unsubstituted aryl, substituted or unsubstituted cycloalkyl, or substituted or unsubstituted heterocyclyl.
  • the present invention also provides the process for making the compounds of formula (I).
  • the present invention further provides a pharmaceutical composition
  • a pharmaceutical composition comprising a therapeutically effective amount of a compound or combination of compounds of the present invention, or a pharmaceutically acceptable salt form, stereoisomer and a pharmaceutically acceptable carrier.
  • W] and ⁇ 2 are independently can be selected from a bond or substituted or unsubstituted alkylene and preferably the substituent is selected from n-butyl, isobutyl or tertiary butyl ;
  • nx and ny are independently can be an integer 0- 1 and with the proviso that one of nx or ny is 1 ;
  • R can be H, -0[C(R a ) 2 ] m -alkyl, -0[C(R b ) 2 ] m -cycloalkyl, -0[C(R c ) 2 ] m - heterocyclyl, substituted or unsubstituted phenyl, substituted or unsubstituted pyridine, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl;
  • R 2 , R 3 , R 2a and R 3a are independently can be selected from H, C(0)R", C(0)OR", or S(0) 2 R";
  • R4 can be H, OH, halogen, NR", C(0)OR", C(0)NR" substituted or unsubstituted alkyl, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl;
  • R5, R6, R7, R 8 , R a, Rea, ?a and R 8a are independently selected from H, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl ;
  • n can be an integer 0-3;
  • R a , R b and R c are independently selected from H, or substituted or unsubstituted alkyl ;
  • R" can be H, substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted cycloalkyl, or substituted or unsubstituted heterocyclyl.
  • the present invention also provides the process for making the compounds of formula (IA).
  • the present invention further provides a pharmaceutical composition
  • a pharmaceutical composition comprising a therapeutically effective amount of a compound or combination of compounds of the present invention, or a pharmaceutically acceptable salt form, stereoisomer and a pharmaceutically acceptable carrier.
  • W] and W 2 are independently can be selected from substituted or unsubstituted alkylene and preferably the substituent is selected from n-butyl, isobutyl or tertiary butyl;
  • Ria can be H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl ;
  • R2, R 3 , R 2a and R 3a are independently can be selected from H, C(0)R", C(0)OR", or S(0) 2 R";
  • R can be H, OH, halogen, NR", C(0)OR", C(0)NR" substituted or unsubstituted alkyl, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl;
  • n can be an integer 1 -3;
  • R" can be H, substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted cycloalkyl, or substituted or unsubstituted heterocyclyl.
  • the present invention also provides the process for making the compounds of formula (EB).
  • the present invention further provides a pharmaceutical composition
  • a pharmaceutical composition comprising a therapeutically effective amount of a compound or combination of compounds of the present invention, or a pharmaceutically acceptable salt form, stereoisomer and a pharmaceutically acceptable carrier.
  • formula (EB) structurally encompasses all stereo isomers, including enatiomers and diastereomers, which may be contemplated from the chemical structure of the genus described herein. Also contemplated are prodrugs of the compounds of the formula (IB), including ester prodrugs.
  • R 2 , R 3 , R 2a and R 3a are independently H, -C(0)CH 3 , -C(0)OCH 3 , -
  • each R 2 , R3, R 2a and R 3a are independently H, and -C(0)OCH 3 .
  • the representative compounds of fromula ( 1 ) which are illustrative in nature only and are not intended to limit to the scope of the invention (Nomenclature has been generated from Chem. Draw Ultra 1 1 .0 version):
  • the compounds and pharmaceutical compositions described herein are useful in the treatment of diseases, conditions and/or disorders mediated by viral infections more particularly for treating, for example, HIV infection .
  • the present patent application further provides a method of treating a disease, condition and/or disorder mediated by viral infections more particularly HIV infection in a subject in need thereof by administering to the subject one or more compounds described herein in the amount effective to treat that infection.
  • the present invention relates to novel aza-peptides containing 2 ,2- disubstituted cyclobutyl and/or substituted alkoxy benzyl derivatives and, a composition for inhibiting Human Immunodeficiency Virus (HIV) and process for making the compounds.
  • HIV Human Immunodeficiency Virus
  • halogen or halo includes fluorine, chlorine, bromine, or iodine.
  • alkyl refers to a straight or branched hydrocarbon chain radical consisting solel y of carbon and hydrogen atoms, containi ng no unsaturation , having from one to eight carbon atoms, and which is attached to the rest of the molecule by a si ngle bond, e.g.. methyl, ethyl , n-propyl , 1 -methylethyl (isopropyl), n-butyl , n- pentyl , and 1 , 1 -dimethylethyl (t-butyl).
  • alkenyl refers to an aliphatic hydrocarbon group containing a carbon-carbon double bond and which may be a straight or branched chain having from 2 to about 10 carbon atoms, e.g., ethenyl , 1 -propenyl, 2-propenyl (allyl), iso- propenyl, 2-methyl- 1 -propenyl, 1 -butenyl, and 2-butenyl .
  • alkylene refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation, having from one to five carbon atoms, and which is attached to the rest of the molecule by two sides bond, e.g., methylene, ethylene, n-propylene, - C(methyl)(isopropyl)-, -CH(isopropyl )-, -CH(n-biityl)-, -CH(t-butyl)-, -CH(iso- butyl)-, -CH 2 -CH(n-butyl)-, -CH 2 -CH(iso-butyl )-, or -CH 2 -CH(t-butyl)-.
  • acyl group is used to denote a linear or branched aliphatic acyl group (preferably a C 2 . 6 alkanoyl group) or an aromatic acyl group, which contains 2 to 10 carbon atoms.
  • examples include an acetyl group, a propionyl group, a pivaloyl group, a butyryl group, an isobutyryl group, a valeryl group and a benzoyl group, with an acetyl group being preferred.
  • alkoxy group is used to denote a linear or branched alkoxy group containing 1 to 6 carbon atoms. Preferred are C
  • alkoxycarbonyl group isused to denote a structure composed of a linear or branched C
  • C2-5 alkoxycarbonyl groups including a methoxycarbonyl group, an ethoxycarbonyl group, a propoxycarbonyl group, an isopropoxycarbonyl group and a butoxycarbonyl group.
  • a methoxycarbonyl group is preferred.
  • cycloalkyl denotes a non-aromatic mono or multicyclic ring system of from 3 to about 1 2 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 .
  • cycloalkylalkyl refers to a cyclic ring-contai ning radical having from 3 to about 8 carbon atoms directly attached to an alkyl group.
  • the cycloalkylalkyl group may be attached to the main structure at any carbon atom in the alkyl group that results in the creation of a stable structure.
  • Non-limiting examples of such groups include cyclopropyl methyl, cyclobiitylethyl , and cyclopentylethyl .
  • 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., -CH2C 6 H 5 and -C2H5C6H5.
  • Substituted refers to 1 -3 substituents on the same position or on different positions with the same groups or different groups.
  • Alkynyl refers to alkynyl groups having from 2 to 6 carbon atoms and having at least one alkynyl saturation, example for such group includes acetylenyl, propargyl.
  • Carbonyloxy refers to a group such as -C(0)0.
  • heterocyclyl'- and “heterocyclic ring” refer to a stable 3- to 15- membered ring radical which consists of carbon atoms and from one to five heteroatoms selected from nitrogen, phosphorus, oxygen and sulfur.
  • the heterocyclic ring radical may be a monocyclic, bicyclic or tricyclic ring system, which may include fused, bridged or spiro ring systems, and the nitrogen, phosphorus, carbon, oxygen or sulfur atoms in the heterocyclic ring radical may be optionally oxidized to various oxidation states.
  • the nitrogen atom may be optionally quaternized; and the ring radical may be partially or fully saturated (i.e., heterocyclic or heteroaryl).
  • heterocyclic ring radicals include, but are not limited to, azetidinyl , acridinyl, carbazolyl, cinnolinyl, indolizinyl, naphthyridinyl, perhydroazepinyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pyridyl, pteridinyl, purinyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrazoyl, imidazolyl, tetrahydroisouinolyl, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2- oxopyrrolidinyl, 2-oxoazepinyl, azepinyl, pyrrolyl, 4-piperidonyl,
  • 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.
  • 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.
  • heteroarylalkyl refers to a heteroaryl ring radical directly bonded to an alkyl group.
  • the heteroarylalkyl 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.
  • R ⁇ R y and R z are independently selected from hydrogen, substituted or unsubstituted alkyl, haloalkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkyl, substituted or un
  • prodrug means a compound that is transformed in vivo to yield a compound of Formula (I) (IA) or ( ⁇ ) or a pharmaceutically acceptable salt, hydrate or solvate 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
  • treating or “treatment” of a state, disease, disorder or condition includes:
  • the benefit to a subject receiving treatment is either statistically significant or at least perceptible to the subject or to the physician.
  • subject includes mammals (especially humans) and other animals, such as domestic animals (e.g., household pets including cats and dogs) and non-domestic animals (such as wildlife).
  • domestic animals e.g., household pets including cats and dogs
  • non-domestic animals such as wildlife.
  • a “therapeutically effective amount” means, for example, 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 compound of the invention may form salts.
  • pharmaceutically acceptable salts forming part of the invention include salts derived from inorganic/organic acids or bases and amino acids salts.
  • Certain compounds of the invention are capable of existing in stereoisomeric forms (e.g. diastereomers and enantiomers). With respect to the overall compounds described by the Formula (I), Formula (LA) or Formula (LB), the invention extends to these stereoisomeric forms and to mixtures thereof. To the extent prior art teaches synthesis or separation of particular stereoisomers, the different stereoisomeric forms of the invention may be separated from one another by the method known in the art, or a given isomer may be obtained by stereospecific or asymmetric synthesis. Tautomeric forms and mixtures of compounds described herein are also contemplated.
  • compositions provided in the 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.
  • 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 bei ngs.
  • the compound of the present invention may be associated with a pharmaceuticall y acceptable excipient (such as a carrier or a diluent) or be diluted by a carrier.
  • 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, pecti n, 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 auxil obviouslyy agents, wetting agents, emulsifyi ng 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 i ngredient after administration to the subject by employing procedures known in the a .
  • the pharmaceutical compositions described herein may be prepared, e.g., as described in Remington: The Science and Practice of Pharmacy, 20 lh Ed., 2003 (Lippincott Williams & Wilkins).
  • compositions may be, for example, capsules, tablets, aerosols, solutions, suspensions 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 preferred.
  • 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. Preferable 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 tabletting techniques may contain: ( 1 ) Core: Active compound (as free compound or salt thereof), colloidal silicon dioxide (Aerosil®), microcrystalline cellulose (Avicel®), modified cellulose gum (Ac-Di-Sol®), and magnesium stearate; (2) Coating: HPMC, Mywacett 9-40 T and acylated monoglyceride.
  • 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 preferably 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.
  • MTT 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazlium bromide
  • 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.
  • 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.
  • Azidothymidine Azidothymidine
  • 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 patent application further provides a method of treating a disease, condition and/or disorder mediated by viral infections in a subject in need thereof by administering to the subject a therapeutically effective amount of a compound or a pharmaceutical composition of the present invention.
  • Diseases, conditions, and/or disorders that are mediated by viral infections are believed to include, but are not limited to, HIV infection, HBV, HCV, a retroviral infection genetically related to HIV, AIDS, or respiratory disorders (including adult respiratory distress syndrome (ARDS).
  • HIV infection HBV
  • HCV a retroviral infection genetically related to HIV
  • AIDS a retroviral infection genetically related to HIV
  • ARDS adult respiratory distress syndrome
  • the compounds of the present invention can also obtain synergistic effects in the prevention or treatment of the above diseases when used suitably in combination with existing drugs.
  • the administered dose may be decreased in comparison with administration of either drug alone, and in addtion adverse effects of co- administrated drugs can be avoided or reduced.
  • 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 4. 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 stereo isomers of the compounds in these schemes, unless otherwise specified, are also encompassed within the scope of this invention.
  • the compounds of Formulae 6 and 13 can be prepared by the above procedure as described in Scheme 1 by adopting the methods published in Tetrahedron :Assymmetry 19(2008302-308) (wherein, P is a protecting group (for example alkyl or aralkylyl protecting groups such as, methyl, ethyl, propyl, isopropyl, benzyl, tertiorybutyl, benzyl carbonyl (Cbz) or tertiorybutyloxy carbonyl
  • P is a protecting group (for example alkyl or aralkylyl protecting groups such as, methyl, ethyl, propyl, isopropyl, benzyl, tertiorybutyl, benzyl carbonyl (Cbz) or tertiorybutyloxy carbonyl
  • Ra and Rb are independently can be selected from H, C(0)R", C(0)OR", or S(0)2R").
  • Commercially available ve benone (formula 1 ) was oxidized with an oxidizing agent like ruthenium chloride/sodium periodate in a solvent system like CC14-ACN-H20 to give the carboxylic acid of compounds of formula 2.
  • the carboxylic acid of compounds of formula 2 is protected with protecting agent to give the acid protected of compounds of formula 3 in the presence of a base like cesium carbonate, or in the presence of a dehydrating agent like DCC or an acid catalyst or the like in the solvents such as DMF, or the like.
  • the acid protected compounds of formula 3 are subjected to Schmidt rearrangement by treatment with sodium azide and methane sulphonic acid or the like to give the N-acyl compounds of formula 4.
  • the N-acyl of compounds of formula 4 was Hydrogenolysed (when P is Bn) in the presence of Pd-C EtOAc or the like to give the N-acyl amino acid of compounds of formula 5.
  • the N-acyl amino acid of compounds of formula 5 can be acid hydrolised in the presence of hydrogen chloride or the like and treated with BOC anhydride in appropriate conditions to give corresponding carbamate more particularly the tertiary butyl carbamate of compounds of formula 6.
  • the carboxylic acid of compounds of formula 2 can be converted in to the azide compounds of formula 7 (through a Curtius rearrangement) in the presence of chloroformates such as for example, ethyl chloroformate or the like and metal azides such as for example, sodium azide or the like in the solvents such as for example, acetone, dichloromethane, dimethylformamide or the like.
  • chloroformates such as for example, ethyl chloroformate or the like
  • metal azides such as for example, sodium azide or the like
  • solvents such as for example, acetone, dichloromethane, dimethylformamide or the like.
  • the azide compounds of formula 7 can be converted to the N-protected compounds of formula 8 as described in J. Org. Chem.
  • N-protected compounds of formula 8 can be converted to the acid compounds of formula 9 by Lieben degradation of methyl ketone function of formula 8 in the presence of dioxane and aqueous sodiumhypobromite or the like.
  • the acid compounds of formula 9 can be converted to the corresponding ester compounds of formula 10 by alkylation of resultant acid in the presence of alkyl halide such as for example, methyl iodide, ethyl iodide or the like and a base such as potassium or cesium carbonate in a solvent like dimethyl formamide.
  • alkyl halide such as for example, methyl iodide, ethyl iodide or the like
  • a base such as potassium or cesium carbonate in a solvent like dimethyl formamide.
  • the alkyl ester compounds of formula 10 can be N- deprotected in the presence of deprotecting agents such as such as for example, palladium on carbon or the like to give the deprotected compounds of formula 1 1 (also Compounds of formula 1 1 can be prepared either from alpha-pinene or verbenone. Alpha-pinene can be converted into verbenone by allylic oxidation).
  • the deprotected compounds of formula 1 1 can be converted to the N-substituted compounds of formula 12 with corresponding chloroformates or the like in the solvents such as, for example, dichloromethane, dimethylformamide and a base like triethyl amine or the like.
  • the N-substituted compounds of formula 12 can be hydrolysed in the presence of base or hydrogenolysed in the presence of Pd- C EtOAc when ester is a benzyl ester or the like to give the acid compounds of formula 13.
  • the compounds of Formula 23 can be prepared by the above procedure as described in Scheme 2 (wherein, P is a protecting group (for example alkyl or aralkylyl protecting groups such as, methyl, ethyl, propyl, isopropyl, benzyl or tertiorybutyl, benzyl carbonyl (Cbz) or tertiorybutyloxy carbonyl (Boc) and the remaining R-groups are same as defined above) by Schotten-Baumann acylation.
  • P is a protecting group (for example alkyl or aralkylyl protecting groups such as, methyl, ethyl, propyl, isopropyl, benzyl or tertiorybutyl, benzyl carbonyl (Cbz) or tertiorybutyloxy carbonyl (Boc) and the remaining R-groups are same as defined above) by Schotten-Baumann acylation.
  • P
  • amino carboxylic acid compounds of formula 14 can be reacted with corresponding substituents for example, chloroformates or the like in the suitable conditions such as, for example, dioxane, sodium hydroxide or the like to give the substituted amino compounds of formula 15.
  • the substituted amino compounds of formula 15 can be reacted with hydrazinecarboxylates to give the protected hydrazine compounds of formula 16 in the presence of coupling agents such as 1 - Ethyl-3-(3-Dimethylaminopropyl)carbodiimide (EDCI), hydroxybenzotriazole (HOBt) or the like in the solvents such as for example, N-methyl morpholine (NMM), dimethylformamide, diiospropyl ethylamine, dichloromethane, ethyl acetate or the like.
  • coupling agents such as 1 - Ethyl-3-(3-Dimethylaminopropyl)carbodiimide (EDCI), hydroxybenzotriazole (HOBt) or the like
  • solvents such as for example, N-methyl morpholine (NMM), dimethylformamide, diiospropyl ethylamine, dichloromethane, ethyl a
  • the protected hydrazine compounds of formula 16 can be deprotected in suitable conditions such as, for example HCl in dioxane, Hal or the like to give the deprotected hydrazine compounds of formula 17.
  • the deprotected hydrazine compounds of formula 17 can be reacted with substituted or unsubstituted benzaldehyde compounds of formula 18 to give the corresponding (benzylidene)hydrazinyl compounds of formula 19 in the presence of alcohols such as, for example, isopropanol, ethanol, methanol or the like.
  • the (benzylidene)hydrazinyl compounds of formula 19 can be hydrogenolysed in the presence of hydrogenolysing agents such as for example, sodiumcyanoborohydride/p-tolunesulfonicacid, palladium on carbon, sodiumformate or the like in the solvents such as ethanol, water or the like to give the corresponding benzyl hydrazine compounds of formula 20.
  • the benzyl hydrazine compounds of formula 20 can be reacted with N-protected amino epoxide compounds of formula 21 to give the corresponding N-protected peptide compounds of formula 22 in the presence of solvents such as for example, isopropanol, ethanol, ethyl acetate or the like.
  • the N-protected peptide compounds of formula 22 can be deprotected by hydrogen chloride in solvents such as dioxane or tetrahydrofuran or the like to give the free amino peptide compounds of formula 23.
  • the compounds of Formula 27 [Formula (I), wherein W
  • protecting groups for example alkyl or aralkylyl protecting groups such as, methyl, ethyl, propyl, isopropyl, benzyl or tertiorybutyl, benzyl carbonyl (Cbz) or tert
  • N-protected benzyl hydrazine compounds of formula 24 can be reacted with N- protected amino epoxide compounds of formula 21 (as described in Bio org. Med. Chem. Lett.1993, 3, 2837-2842) to give the corresponding N-protected peptide compounds of formula 25 in the presence of solvents such as for example, isopropanol, ethanol, ethyl acetate or the like.
  • solvents such as for example, isopropanol, ethanol, ethyl acetate or the like.
  • the N-protected peptide compounds of formula 25 can be deprotected by hydrogen chloride in the solvents such as
  • Ethyl-3-(3-Dimethylaminopropyl)carbodiimide EDCI
  • HOBt hydroxybenzotriazole
  • solvents such as for example, N-methyl morpholine (NMM), dimethylformamide, diiospropyl ethylamine, dichloromethane, ethyl acetate or the like.
  • Step 1 synthesis of ( lS,3R)-3-acetyl-2,2-dimethylcyclobutanecarbo.xylic acid:
  • reaction mixture was diluted with EtOAc, filtered through celite bed and the organic layer was separated then the aqueous layer was extracted with EtOAc ( 100 ml x 2). Combined organic layers were dried with Na 2 S0 4 and concentrated under reduced pressure. The resulting crude was stirred in hexane, forming the solid. Hexane was decanted, and the resulting solid was dried under vacuum.
  • reaction mixture was diluted with EtOAc (100 ml) and washed with water (50 ml), brine (50 ml) and dried with Na 2 S04, the solvent was evaporated and purified by silica gel column (60-120, elution 4% EtOAc in hexane) chromatography to afford the title compound as a pale yellow syrup.
  • Step 2 Synthesis of benzyl ( lS,3R ethylcyclobut ⁇ lcarbaniate:
  • NaOBr Solution A solution of NaOH (about 9.4 g, 233.6 mmol) in H 2 0 ( 180 ml) Br 2 (3.3 ml, 63.72 mmol) was added at about 0°C slowly then the yellow colored NaOBr solution formed which was used immediately for the reaction.
  • Step 1 Synthesis of ( I R,3S)-methyl 3-(benzyloxycarbonylamino)-2,2- dimethylcyclob tanecarboxylate:
  • Step 2 Synthesis of ( 1 R,3S)-methyI 3-(metho.xycarbonylamino)-2,2- dimethylcyclobntanecarboxylate:
  • step 1 A stirred solution of ( l R,3S)-methyl 3-(benzyloxycarbonylamino)-2,2- dimethylcyclobutanecarboxylate (step 1 , about 1.0 g, 3.436 mmol) in EtOH ( 15 ml) at room temperature 10 % Pd/C (catalytic amount) was added and the reaction mixture was stirred under H 2 gas atmosphere at same temperature for about 12 hours.
  • Step 3 Synthesis of ( lR,3S)-3-(methoxycarbonyla ino)-2,2- dimethylcyclobutanecarboxylic acid:
  • Step 1 Synthesis of (lR,3S)-methyl 3-(tert-butoxycarbony ⁇ amino)-2,2- dimethylcyclobutanecarboxylate:
  • CD 3 OD ⁇ 3.78-3.69 (m, 1 H), 3.65 (s, 3H), 2.66-2.57 (m, 1 H), 2.24-2. 10 (m, 2H), 1 .43 (s, 9H), 1 .24 (s, 3H), 0.86 (s, 3H);
  • IR KBr, cm “ 1 ): 3404, 3384, 2957, 1706, 1522, 1462, 1439, 1367, 1343, 1266, 1 165, 1008, 984, 871 , 778; Mass: [M+Na] + 280 ( 100%); M R: 101 .2- 103.5 °C.
  • Step 2 Synthesis of ( 1 R,3S)-3-(tert-butoxycarbonylainino)-2,2- dimethyicyclobutanecarboxylic acid:
  • step 1 To a stirred solution of ( l R,3S)-methyl 3-(tert-butoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylate (step 1 , about 1 .5 g, 5.836 mmol) in tetrahydrofuran (60 ml) at about 0 °C sodium hydroxide (0.25 M, 46.6 ml) was added and stirred at room temperature for about 2 hours. After completion of the reaction (monitored by TLC), the solvent was evaporated, acidified with citric acid and extracted with EtOAc.
  • Step 1 Synthesis of tert -butyl 2-((2S,3S)-3-(tert-butoxycarbonylamino)-2-hydroxy-4- phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hydrazinecarboxylate:
  • step 1 To a stirred solution of tert-butyl 2-((2S,3S)-3-(tert-butoxycarbonylamino)-2- hydroxy-4-phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hydrazinecarboxylate (step 1 , about 0.5 g, 0.889 mmol) in tetrahydrofuran (2.5 ml) concentrated HCl (0.339 ml) was added drop wise at room temperature to give colored solution. The reaction mixture was stirred at about 45-55 0°C for about 3 hours. After completion of the reaction tetrahydrofuran layer was removed and the remaining oil compound rinsed with tetrahydrofuran ( 1 ml) then it was used as such for further reactions.
  • Step 1 Synthesis of (S)-2-(methoxycarbonylaniino)-3,3-dimethylb tanoic acid:
  • step 1 To a stirred solution of (S)-2-(methoxycarbonylamino)-3,3-dimethylbutanoic acid (step 1 , about 8.50 g, 44.97 mmol) in EtOAc ( 1 10 ml) at about 0 °C l -Ethyl-3- (3-Dimethylaminopropyl)carbodiimide (9.48 g, 49.47 mmol), hydroxybenzotnazole (7.57 g, 49.47 mmol) and N-methyl morpholine (5.95 ml, 53.95 mmol) were added, after 30 minutes t-Butyl carbazate (about 6.53 g, 1 .423 mmol) was added and stirred at room temperature for about 18 hours.
  • Step 3 Synthesis of (S)-niethyl 1 -hydrazinyl-3, 3 -dimethyl- 1 -oxob tan-2- xlcarbamate:
  • Step 4 synthesis of (S,E)-methyl 3,3-di ethyl-l -oxo-l -(2-(4-(pyridin-2- yl)benzylidene)hydrazinyl)buta -2-ylcarbamate: A solution of (S)-methyl l -hydrazinyl-3,3-dimethyl- l -oxobutan-2- ylcarbamate (step 3, about 1.56 g, 7.722 mmol) and 4-(pyridin-2-yl)benzaldehyde (about 1 .407 g, 7.722 mmol) in isopropyl alcohol stirred at reflux temperature for about 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated and purified by silica gel column ( 100-200 mesh, elution
  • Step 5 Synthesis of (S)-methyl 3,3-dimethyl-l -oxo- l-(2-(4-(pyridin-2- yl)benzyl)hydrazinyl)butan-2-ylcarbamate:
  • step 4 To a solution of (S,E)-methyl 3,3-dimethyl- l -oxo- l -(2-(4-(pyridin-2- yl)benzylidene)hydrazinyl)butan-2-ylcarbamate (step 4, about 3.0 g, 8. 16 mmol) in EtOH (20 ml) sodiumformate (about 0.99 g, 14.673 mmol) was dissolved in minimum amount of water and palladium on carbon ( 10%) were added and stirred under H 2 gas atmosphere at room temperature for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through celite bed and the obtained filtrate was concentrated under reduced pressure.
  • Step 6 Synthesis of methyl (S)-l -(2-((2S,3S)-3-(tert-butoxycarbonylamino)-2- hydrox)'-4 ⁇ henylbuty1)-2-(4-(pyridin-2-yl)benzyl)hydrazinyl)-3 -dimethy oxobutan-2-ylcarbamate:
  • Step 7 Synthesis of methyl (S)-l -(2-((2S,3S)-3-amino-2-hydroxy-4-phenylb tyl)-2- (4-(pyridin-2-yl)benzyl)hydraz.inyl)-3, 3 -dimethyl- 1 -oxobittan-2-y /carbamate:
  • Step 1 Synthesis of (S,E)-methyl l-(2-benzylidenehydrazinyl)-3,3-dimethyl-l- oxob tan-2-ylcarbamate: 0
  • Step 2 Synthesis of (S)-methyl 1 -(2-benzylhydrazinyl)-3,3-dimethyl-J -oxobutan-2- vlcarbamate
  • step 1 A solution of (S,E)-methyl l -(2-benzylidenehydrazinyl)-3,3-dimethyl- l - oxobutan-2-ylcarbamate (step 1 , about 2.4 g, 8.24 mmol) in EtOH (40 ml), HCCbNa ( 1 .0 g, 14.84 mmol) was dissolved in minimum amount of water and palladium on carbon ( 10%) were added and stirred under H? gas atmosphere at room temperature for about 12 hours. After completion of the reaction (monitored by TLC) the reaction mixture was filtered through celite bed and the obtained filtrate was concentrated under reduced pressure.
  • Step 3 Synthesis of methyl (S)-l -(2-benzyl-2-((2S S)-3-(tert- b ⁇ toxycarbonylami o)-2 ⁇ ydroxy-4 ⁇ henylb ⁇ t ⁇ 1)hydrazinyl)-3 -dimethyl- l - oxobu tan -2 -y lea rba mate:
  • Step 4 Synthesis of methyl (S)-l -(2-((2S,3S)-3-amino-2-hydroxy-4-phenylbutyl)-2- benzylhydrazinyl)-3, 3 -dimethyl- 1 -oxobutan-2-ylcarbamate :
  • Step 1 Synthesis of biphenyl-4-carbaldehyde :
  • Step 2 Synthesis of (S,E)-methyl l -(2-(biphenyI-4-ylmethylene)hydrazinyl)-3,3- dimethyl-l -oxobutan-2-ylcarbamate:
  • Step 3 synthesis of (S)-methyl l -(2-(biphenyl-4-ylmethyl)hydrazinyl)-3,3-dimethyl- l -oxobutan-2-xlcarbaniate: To a stirred solution of (S,E)-methyl l -(2-(biphenyl-4- ylmethylene)hydrazinyl)-3,3-dimethyl- l -oxobutan-2-ylcarbamate (step 2, about 2.7 g, 7.356 mmol) in ethanol (30 ml) HC0 2 Na (0.90 g, 13.24 mmol) dissolved in minimum amount of water and Pd/C ( 10%) were added and stirred under H? gas atmosphere at room temperature for about 2 hours. After completion of the reaction
  • Step 4 Synthesis of methyl ⁇ S)-l-(2-(biphenyl-4-yhnethyl)-2-((2S, 3S)-3-(tert- butoxycarbony ⁇ amm ' o)-2-hydroxy-4-phenylbuty ⁇ )hydraz ⁇ ny ⁇ )-3
  • Step 5 Synthesis of methyl (S)-l -(2-((2S,3S)-3-amino-2-hydroxy-4-phenylbutyl)-2- ( biphenyl-4-ylmethyl)hydrazinyl)-3, 3 -dimethyl- 1 -oxobutan-2-ylcarbamate:
  • oxob tan-2-ylcarbamate (Step 4, about 0.3 g, 0.474 mmol) in tetrahydrofuran (6.0 ml) at about °C 4N HCl (4 ml) was added drop wise and stirred at about 50 °C for about 2 hours. After completion of the reaction the solvent was removed and dissolved in EtOH then concentrated and dried under vacuum then it was used as such for further reactions.
  • Step 1 Synthesis of ( S )-2 -( methoxycarbonylamino)-4-methylpentonoic acid:
  • Step 2 Synthesis of (S)-tert-b t)'l 2-(2-(methoxycarbonylamino)-4-methylpentanoyl) hydrazinecarboxylate :
  • step 1 To a stirred solution (S)-2-(methoxycarbonylamino)-4-methylpentanoic acid (step 1 , about 8.0 g, 42.328 mmol) in EtOAc (50 ml) at about 0 °C l -Ethyl-3-(3- Dimethylaminopropyl)carbodiimide (8.925 g, 46.560 mmol), hydroxybenzotriazole (7. 13 g, 49.560 mmol) and NMM (5.6 ml, 50.793 mmol) were added and stirred for 30 minutes then t-Butyl carbazate (6. 152 g, 46.560 mmol) was added and stirred at room temperature for 18 hours.
  • Step 3 synthesis of (S)-methyl 1 -hydrazinyl-4-methyl- 1 -oxopentan-2-xlcarbamate :
  • step 2 A solution of (S)-tert-butyl 2-(2-(methoxycarbonylamino)-4- methylpentanoyl)hydrazinecarboxylate (step 2, about 1 .0 g, 3.30 mmol) in 2N HC1 and dioxane (20 ml) at about 0°C was stirred and allowed to reach room temperature, then continued stirring for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated, basified with solid NaHC0 , extracted with EtOAc and dried over Na 2 S04 and the solvent was evaporated and the resulting crude was proceeded for next reaction without characterization. Wt: 0.66 g: Yield: 100 %.
  • Step 4 Synthesis of (S, E)-methyl 4-methyl-l -oxo-l -(2-(4-(pyridin-2-
  • Step 5 Synthesis of (S)-methyl 4-methyl-l -oxo-l-(2-(4-(pyridin-2- yl)benzyl)hydrazinyl )pentan-2-ylcarbamate:
  • Step 6 Synthesis of (S) methyl -l-(2-((2S,3S)-3-(tert-butoxycarbonylamino)-2- hydrox -4 ⁇ henylbut ⁇ 'l)-2-(4-(pyridin-2-yl)benzyl)hydrazinyl)-4-methyl-l- oxopentan-2-ylcarbamate :
  • Step 7 Synthesis of methyl (S)-l-(2-((2S3S)-3-amino-2-hydroxy-4-phenylb tyl)-2- (4-(pyridin-2-yl)benzyl)hydrazinyl)- -methyl-l-oxopentan-2-ylcarbainate:
  • Step 2 Synthesis of 4-(2-(4-ethylpiperazin-l-yl) ethoxy) benzaldehyde:
  • Step 3 Synthesis of (R, Z)-methyl l-(2-(4-(2-(4-ethylpiperazin-l -yl) ethoxy) benzylidene) hydrazinyl)-3, 3-dimethyl-l -oxob tan-2-ylcarbamate:
  • reaction mixture was refluxed for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated, diluted with ethyl acetate, washed with water, brine solution and dried over Na2S04 then concentrated under reduced pressure and purified by column chromatography using hexane and ethyl acetate mixture as an eluent to afford the tittle compound (4.0 g) yield 37 %.
  • Step 4 Synthesis of (R)-methyl l -(2-(4-(2-(4-ethylpiperazin-l -yl) ethoxy) benzyl) hydrazinyl)-3, 3 -dimethyl- 1 -oxobutan-2-ylcarbamate:
  • Step 1 Synthesis of 4-(2-(3, 4-dihydroquinolin- 1 (2H )-yl) ethoxy) benzaldehyde:
  • Step 2 Synthesis of (R, Z)-methyl l-(2-(4-(2-(3, 4-dihydroquinolin-l(2H)-yl) ethoxy) benzylidene) hydrazinyl)-3, 3 -dimeth - l -oxobutan-2-ylcarbamate:
  • Step 3 Synthesis of (R)-methyl l-(2-(4-(2-(3, 4-dihydroquinolin-l(2H)-yl) ethoxy) benzyl) hydrazinyl)-3, 3 -dimethyl- l -oxobutan-2-ylcarbamate:
  • Step 2 and step 3 procedure are same as in Intermediate 13 to get the Intermediate 14.
  • Step 3 Synthesis of methyl N-[(lS)-2,2-diinethyl-]- ⁇ N'-[(JE)-(4-[2-(morpholin-4- yljethoxy] phenyl ⁇ methylidene]hydrazinecarbonylj propyl] carbamate:
  • Step 4 Synthesis of methyl N-[( lS)-2,2-dimethyl-l -[N'-( ⁇ 4-[2-(morpholin-4- yljethoxy] phenyl ⁇ methyl)hydrazinecarbony I] propyl] carbamate:
  • Example 1 Preparation of (lS,3R)-3-acetamjdo-N-((2S,3S)-4-(2-((lS,3R)-3- acetamido-2,2-dimethylcyclobutanecarbonyl)-l-(4-(pyridin-2-yl)benzyl)hydrazinyl)- 3-hydroxy-l-phenylbutan- -yl)-2,2-dimethylcvclobutanecarboxamide:
  • Example 2 Preparation of methyl ( l R,3S )-3-((2S,3S)-4-(2-(( l S,3R)-3- methylcarbamato-2,2-dimethylcyclobutanecarbonyl)- l -(4-(pyridin-2- yl)benzyl)hydrazinyr)-3-hydroxy- l -phenylbutan-2-ylcarbamoyl )-2,2- dimethylcyclobutylcarbamate:
  • Example 3 Preparation of ten-butyl (lS,3R>-3-((2S,3S)-4-(2-((lR.3S)-3-(tert- butyloxycarbonylamino)-2,2-dimethylcyclobutanecarbonyl)-l-(4-(pyridin-2- yl)benzyl)hydrazinyl)-3-hydroxy-l-phenylbutan-2-ylcarbamoyl)-2,2- dimethylcyclobutylcarbamat
  • Dimethylaminopropyl)carbodiimide (about 1.04 g, 5.685 mmol) were added at about 0 °C, after 10 minutes (2S,3S)-3-amino-4-phenyl-l-(l-(4-(pyridin-2- yl)benzyl)hydrazinyl)butan-2-ol (Intermediate 7, about 0.671 g, 1.423 mmol) in DCM (1 ml), DMF (0.5 ml) and diiospropyl ethylamine (about 1.2 ml, 7.115 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 16 hours.
  • Example 4 Preparation of benzyl (lS,3R)-3-((2S.3S)-4-(2-((lR,3S)-3- (benzyloxycarbonylamino)-2,2-dimethylcvclobutanecarbonyr)-l-(4-(pyridin-2- yl)benzyl)hydrazinyl)-3-hydroxy-l-phenylbutan-2-ylcarbamoyD-2,2- dimethylcyclobutylcarbama
  • Example 5 Preparation of methyl ( l S R)-3-((2S.3S)-3-hvdroxy-4-(2-(( l R.3S)-3- (methoxycarbonylamino)-2,2-dimethylcyclobutanecarbonyl)- l -(4-(pyridin-2- yPbenzyPhydrazinyl )- 1 -phenylbutan-2-ylcarbamoyl)-2,2- dimethylcyclobutylcarbama
  • Example 7 Preparation of methyl (S)-l-(2-((2S.3S)-3-((lS,3R)-3-(tert- butoxYcarbonylamino)-2,2-dimethylcvclobutanecarboxamido)-2-hvdiOxy-4- phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2- ylcarbamate:
  • reaction mixture was diluted with DCM and washed with water, saturated NaHCC solution and brine.
  • the organic layer was concentrated under reduced pressure and the resulting crude was purified by silica gel column chromatography (100-200 mesh, elution 35% EtOAC in hexane) to afford the title compound as a white solid.
  • Example 8 Preparation of methyl (S)-l-(2-((2S,3S)-3-((lS,3R)-3-acetamido-2,2- dimethylcvclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-(4-(pyridin-2- yl)benzyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate:
  • Example 9 Preparation of methyl (S)-l-(2-((2S,3S)-2-hydroxy-3-((lR,3S)-3- (methoxycarbonylamino)-2,2-dimethylcyclobutanecarboxamido)-4-phenylbutyl)-2- (4-(pyridin-2-yl)benzyl)hvdrazinyr)-3,3-dimeth l-l-oxobutan-2-ylcarbamate:
  • Example 10 Preparation of methyl (S)-l-(2-((2S,3S)-3-((lR,3S)-3-(tert- butoxycarbonylamino)-2,2-dimethylcvclobutanecarboxamjdo)-2-hydroxy-4- phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hvdrazinyl )-3,3-dimethyl- 1 -oxobutan-2- ylcarbamate:
  • reaction mixture was diluted with DCM and washed with water, saturated NaHC0 3 solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100-200 mesh, elution 45% EtOAc in hexane) to afford the title compound as an off white solid.
  • Example 11 Preparation of methyl (S)-l-(2-((2S,3S)-3-((lR,3S)-3- (benzyloxycarbonylamino)-2,2-dimethylcyclobutanecarboxamido)-2-hydroxy-4- phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hvdrazinyr)-3,3-dimethyl-l-oxobutan-2- ylcarbamate:
  • reaction mixture was diluted with DCM and washed with water, saturated NaHC0 3 solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100-200 mesh, elution 45% EtOAC in hexane) to afford the title compound as an off white solid.
  • Example 12 Preparation of methyl (S)-l-(2-benzyl-2-((2S,3S)-2-hydroxy-3- (( lS,3R)-3-(methoxycarbonylarnino)-2,2-dimethylcyclobutanecarboxamido)-4- phenylbutyl)hvdrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate:
  • reaction mixture was diluted with DCM and washed with water, saturated NaHC0 3 solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100-200 mesh, elution 55% EtOAc in hexane) to afford the title compound as a white solid.
  • Dimethylaminopropyl)carbodiirrude (about 0.258 g, 1.347 mmol) were added at about 0 °C for about 10 minutes then methyl (S)-l-(2-((2S,3S)-3-amino-2-hydroxy- 4-phenylbutyl)-2-benzylhydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate (Intermediate 9, about 0.409 g, 0.898 mmol) in DCM (1 ml), DMF (1 ml) and NMM (about 0.5 ml, 4.49 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 42 hours.
  • reaction mixture was diluted with DCM and washed with water, saturated NaHCCh solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100-200 mesh, elution 90% EtOAc in hexane) to afford the title compound as a white solid.
  • Example 14 Preparation of methyl (S)-l-(2-benzyl-2-((2S,3S)-3-((lS.3R)-3-(tert- butoxycarbonylamino)-2,2-dimethylcyclobutanecarboxamido)-2-hydroxy-4- phenylbutyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate:
  • Example 15 Preparation of methyl (S)-l-(2-((2S.3S)-3-((lS.3R)-3-acetamido-2,2- dimethylcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyr)-2-(biphenyl-4- ylmethyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate:
  • Example 16 Preparation of methyl (S)- l -(2-(biphenyl-4-ylmethyl)-2-((2S,3S)-2- hydroxy-3-(( 1 S,3R)-3-(methoxycarbonylamino)-2,2- dimethylcyclobutanecarboxamido)-4-phenylbutyl)hydrazin yl )-3,3-dimethyl- 1 - oxobutan-2-ylcarbamate:
  • reaction mixture was diluted with DCM and washed with water, saturated NaHC0 3 solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100-200 mesh, elution 50% EtOAC in hexane) to afford the title compound as a white solid.
  • Example 18 Preparation of methyl (S)-l-(2-(biphenyl-4-ylmethyl)-2-((2S,3S)-2- hydroxy-3-(( 1 R,3S)-3-(methoxycarbonylamino)-2,2- dimethylcyclobutanecarboxamido)-4-phenylbutyl)hydrazinyl)-3,3-dimethyl-l- oxobutan-2-ylcarbamate:
  • Example 20 Preparation of methyl (S)-l-(2-((2S,3S)-3-((lR,3S)-3- (benzyloxycarbonylamino)-2,2-dimethylcyclobutanecarboxamido)-2-hydroxy-4- phenylbutyl)-2-(biphenyl-4-ylmethvDhvdrazinyl)-3,3-dimethyl-l-oxobutan-2- ylcarbamate:
  • Example 21 Preparation of methyl (S)-l-(2-((2S,3S)-2-hvdroxy-3-((lS.3R)-3- (methoxycarbonylarruno)-2,2-dimethylcyclobutanecarboxarnido)-4-phenylbutyl)-2- (4-(pyridin-2-yl)benzyl)hvdrazinyl)-4-methyl-l-oxopentan-2-ylcarbamate:
  • Example 23 Preparation of methyl (S)-l-(2-((2S,3S)-3-((lS.3R -3-acetamido-2,2- dimethylcvclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-(4-(pyridin-2- yl)benzyl)hydrazinyl)-4-methyl- 1 -oxopentan-2-ylcarbamate:
  • the reaction mixture was diluted with DCM and washed with water, saturated NaHC0 3 solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100- 200 mesh, elution 90% EtOAC in hexane) and the obtained solid was recrystalized from DCM-hexane, MeOH-water to afford the title compound as an off white solid.
  • Example 25 Preparation of methyl (S)-l-(2-((2S.3S)-3-((lR,3S)-3-(tert- butoxycarbonylamino)-2,2-dimethylcyclobutanecarboxamido)-2-hvdroxy-4- phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hvdrazinyl)-4-methyl-l-oxopentan-2- ylcarbamate:
  • reaction mixture was diluted with DCM and washed with water, saturated NaHCCh solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100-200 mesh, elution 90% EtOAC in hexane) to afford the title compound as an off white solid.
  • Step 1 Synthesis of methyl N-[(lS)-l-(N'-[(2S l 3S)-3-([(tert- butoxy)carbonyl)amino ⁇ -2-hydroxy-4-phenylbutyl]-N'- ⁇ 4-[2-(morpholin-4- yl )ethoxy) phenyl Jmethy I )h hy I propyl) carbamate:
  • Step 2 Synthesis of methyl N-[(lS)-l- ⁇ N'-[(2S,3S)-2-hydroxy-3-l(2S)-2- [(methoxycarbonyl)ainino]-3,3-dimethylbi anamido]-4-phenylbut ⁇ 'l]-N'-( ⁇ 4-[2- (morpholin-4-yl)ethoxy] phenyl jmethyl)hydrazinecarbonylj -2,2- dimethylpropyl] carbamate :
  • Example 30 Methyl N-r(lS)-l-fN'-benzyl-N'-f(lS,2R)-2-if(lS,3R -3-(f(tert- butoxy)carbonyllamino ) -2,2-dimethylcyclobutyllformamido ) - 1 -hydroxy-3- phenylpropyllhvdrazinecarbonyl)-2,2-dimethylpropyllcarbamate:
  • Example 31 Methyl N-r(lR.3S)-3-U(lS,2R)-l-hvdiOxy-l- ⁇ l-f(lR,3S)-3- [(methoxycarbonyl)aminol-2,2-dimethylcyclobutyn-N'-( [4-(pyridin-2- yPphenyllmethyl )formohydrazido)-3-phenylpropan-2-yllcarbamoyl 1-2,2- dimethylcyclobutyllcarbamat
  • Example 32 Methyl N-f( 1 S )- 1 - ( N'-f( 1 S.2RV2- f K 1 S.3R>-3- ( idert- butoxy)carbon yllamino ) -2,2-dimethylcyclobutyllformamido ) - 1 -hydiOxy-3- phenylpropyn-N'-( ⁇ 4-f2-(morpholin-4- yDethoxylphenyl ) methyl )hydrazinecarbonyl )-2,2-dimethylpropyllcarbamate:
  • Example 33 Methyl N-I(lS)-l-(N'-f(lS.2R)-2-(f(lR.3S)-3-(i(tert- butox y)carbonyllamino ) -2,2-dimethylcvclobutyllformarnido ) - 1 -h ydroxy-3- phenylpropyll-N'-( ( 4-[2-(morpholin-4- ypethoxylphenyl ImethyPhydrazinecarbonyl 1-2.2-dimethylpropyllcarbamate:
  • Example 34 Methyl N-r(lS)-l-(N'-r(lS,2R)-l-hvdroxy-2-(i(lS,3R)-3- f(methoxycarbonyl)amino1-2,2-dimethylcvclobutynformamido)-3-phenylpropyll- N'-( ( 4-f 2-(morpholin-4-yl)ethoxy1phenyl ) meth vDhvdrazinecarbonyl ⁇ -2,2- dimethylpropyllcarbamate:
  • Example 35 Preparation of methyl N-i(lR)-l-(i(lS.2R)-l-hydroxy-l-f(2S)-2- f (methoxycarbonyl )amino1-3,3-dimethyl-N'-( f 4-f 2-( 1 ,2,3,4-tetrahydroquinolin- 1 - yl)ethoxylphenyl)methyl)butanehydrazidol-3-phenylpropan-2-yllcarbamoyl 1-2,2- dimethylpropyllcarbamate:
  • Step 1 Synthesis of methyl N- ⁇ ⁇ -[ ⁇ '-(3- ⁇ [(tert-butoxy )carbonyl] amino ⁇ -2-hydr
  • Example 36 Preparation of methyl N-f(lR)-l-i iaS,2R)-l-i(2S)-N'-((4-i2-(4- ethylpiperazin-l-yl)ethoxylphenyl )methyl)-2-f(methoxycarbonyl)arrunol-3,3- dimethylbutanehydrazidol-l-hvdroxy-3-phenylpropan-2-yllcarbamoyl)-2,2- dimethylpropyllcarbamate:
  • Step 1 Synthesis of methyl N- ⁇ 1 -[N'-(3- ⁇ [(tert-b toxy)carbonyl]cunino ⁇ -2-hydro 4-phenylb tyl)-N'-( ⁇ 4-[2-(4-ethylpiperazin-l- yl)ethoxy] phenyl ⁇ methyl)hydrazinecarbonyl) -2, 2-dimethylpropylj 'carbamate:
  • Step 2 Synthesis of methyl N-[( 1 R)-l -/ [( 1 S,2R)-J -((2S)-N'-((4-[2-(4-ethylpiperazin- 1 -yljethoxy] phenyl jmethyl)-2f(methoxycarbonyl)amino] -3,3- dimethylbutanehydrazido] - 1 -hydroxy -3-pheny ⁇ propan-2-yl] 'carbamoyl ⁇ '-2,2- dimethylpropyl] carbamate
  • Example 37 Methyl N-f( l R )- l - ( f( l S.2R )- l -hvdroxy- l -f(2S)-2- [(methoxycarbonvDaminol-3,3-dimethyl-N'-( ( 4-[3-( 1 ,2,3,4-tetrahydroquinolin- 1 - yDpropoxylphenyl ) methyl )butanehydrazidol-3-phenylpropan-2-yllcarbamoyl ) -2,2- dimethylpropyllcarbamate:
  • Example 38 Methyl N-f( lS)-l-IN'-i(3S)-3-i(2S)-2-( r(tert-butoxy)carbonyllamino)- 4-methylpentanamidol-2-hydroxy-4-phenylb ⁇ ltyll-N'-((4-[2-(mo holin-4- yl)ethoxy]phenyl )methyl)hydrazinecarbonyl)-2,2-dimethylpropyllcarbamate:
  • Example 39 Screening the activity of antivirals using Cell viability assay:
  • MTT 3-(4,5-dimethylthiozol-2-yl)-2,5-diphenyltetrazolium bromide
  • Example 40 Screening the activity of antiviral :
  • MT2 cells were infected with HIV- 1 strain 92HT599 ( 10 TCID 50/30000 cells).
  • the infected cells were plated at the concentration of -30000 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).
  • cytotoxicity assay For cytotoxicity assay the same amount of MT2 cells as in antivirus without HrV- 1 virus was added to the cytotoxicity plates. The cytotoxicity was measured using MTT reagent in parallel with P24 estimation. The percent viability is calculated in comparison with vehicle control.

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Abstract

The present invention relates to novel aza-peptides containing 2,2-disubstituted 5 cyclobutyl and/or substituted alkoxy benzyl derivatives of formula (I) and compositions for inhibiting Human Immunodeficiency Virus (HIV) and process for making the compounds.

Description

NOVEL AZA-PEPTIDES CONTAINING 2,2-DISUBSTITUTED CYCLOBUTYL AND/OR SUBSTITUTED ALKOXY BENZYL
DERIVATIVES AS ANTIVIRALS This application claims the benefit of Indian Provisional Patent Application
Nos. 3210/CHE/2009 filed on 29th December 2009, which is incorporated herein by reference.
Field of the Invention
The present invention relates to novel aza-peptides containing 2,2- disubstituted cyclobutyl and/or substituted alkoxy benzyl derivatives and compositions for inhibiting Human Immunodeficiency Virus (HIV) and process for making the compounds.
Background of the Invention
HIV- 1 infection remains a major medical problem, with an estimated 34 million people infected worldwide. The number of cases of HIV and AIDS (acquired immunodeficiency syndrome) has risen rapidly. In 2009. approximately 2.6 million new infections were reported, and 2.0 million people died from AIDS.
HIV protease inhibitors are one important class of therapeutic agents for inhibition and treatment of HIV infection. Several protease inhibitors like saquinavir, ritonavir, indinavir, nelfinavir, amprenavir, lopinavir, fosamprenavir and atazanavir have been approved as drugs for treatment of HIV infection. There is a continuing need for novel HIV inhibitors that are very potent and effective against resistant strains of HIV. Therefore, novel anti-HIV agents exhibiting distinct resistance patterns, and favorable pharmacokinetic as well as safety profiles are needed to provide more treatment options.
Aza-peptide structural classes of compounds as HIV- protease inhibitors were explored intensively for their therapeutic activities and those include for example, the patent publications: WO 97/40029, US0584991 1 and US06087383 disclosed antivirally active heterocyclic azahexane derivatives; WO 94/ 19332 disclosed retroviral protease inhibiting compounds; WO 97/ 19055 disclosed azahexane derivatives as substrate isosters of retroviral aspirate proteases; WO 98/03476 disclosed anilinopeptide derivatives; and WO 08/ 1 56632 disclosed azapeptide derivatives. However no patent or publication in the literature has reported the specific compounds or their anti HIV activity of the present invention. This observation is of practical importance in synthesizing the compounds of the present invention and screening them for their anti-HIV activity, for the discovery of new drugs.
Summary of the Invention
The present patent application relates to compounds of the formula (I):
Figure imgf000003_0001
Formula «,!)
or a pharmaceutically acceptable salt, prodrug or stereoisomer there of, wherein,
Wj and W2 are independently selected from a bond or substituted or unsubstituted alkylene and preferably the substituent is selected from n-butyl, isobutyl or tertiary butyl ;
Zi and Z2 are independently selected from a bond or substituted or unsubstituted cyclo butyl ; provided that when atleast one of the Zi and Z2 are substituted or unsubstituted cyclo butyl then Ri can be H, -0[C(Ra)2]m-alkyl, - 0[C(Rb)2]m-cycloalkyl, -0[C(Rc)2]m-heterocyclyl, substituted or unsubstituted phenyl, substituted or unsubstituted pyridine, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl; or when both Zi and Z2 are a bond then Ri can be -0[C(Ra)2]n-Ria > wherein Ria can be H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl;
R2, R3, R2a and R3a are independently can be selected from H, C(0)R", C(0)OR", or S(0):R";
R can be H, OH, halogen, NR", C(0)OR", C(0)NR" substituted or unsubstituted alkyl, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl;
m can be an integer 0-3;
n can be an integer 1 -3 ;
Ra, Rb and Rc are independently can be selected from H, or substituted or unsubstituted alkyl ; R" can be H, substituted or unsubstituted alky], substituted or unsubstituted aryl, substituted or unsubstituted cycloalkyl, or substituted or unsubstituted heterocyclyl.
The present invention also provides the process for making the compounds of formula (I).
The present invention further provides a pharmaceutical composition comprising a therapeutically effective amount of a compound or combination of compounds of the present invention, or a pharmaceutically acceptable salt form, stereoisomer and a pharmaceutically acceptable carrier.
Pharmaceutically acceptable salts of the compounds of the formula (I) are also contemplated.
It should be understood that the formula (I) structurally encompasses all stereo isomers, including enatiomers and diastereomers, which may be contemplated from the chemical structure of the genus described herein.
Also contemplated are prodrugs of the compounds of the formula (I), including for example ester prodrugs.
Accordingly, one aspect of the present invention provides compounds of formula (IA):
Figure imgf000004_0001
Formula ( IA)
or a pharmaceutically acceptable salt, prodrug or stereoisomer there of, wherein,
W] and \ν2 are independently can be selected from a bond or substituted or unsubstituted alkylene and preferably the substituent is selected from n-butyl, isobutyl or tertiary butyl ;
nx and ny are independently can be an integer 0- 1 and with the proviso that one of nx or ny is 1 ;
R, can be H, -0[C(Ra)2]m-alkyl, -0[C(Rb)2]m-cycloalkyl, -0[C(Rc)2]m- heterocyclyl, substituted or unsubstituted phenyl, substituted or unsubstituted pyridine, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl;
R2, R3, R2a and R3a are independently can be selected from H, C(0)R", C(0)OR", or S(0)2R";
R4 can be H, OH, halogen, NR", C(0)OR", C(0)NR" substituted or unsubstituted alkyl, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl;
R5, R6, R7, R8, R a, Rea, ?a and R8a are independently selected from H, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl ;
m can be an integer 0-3;
Ra, Rb and Rc are independently selected from H, or substituted or unsubstituted alkyl ;
R" can be H, substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted cycloalkyl, or substituted or unsubstituted heterocyclyl.
The present invention also provides the process for making the compounds of formula (IA).
The present invention further provides a pharmaceutical composition comprising a therapeutically effective amount of a compound or combination of compounds of the present invention, or a pharmaceutically acceptable salt form, stereoisomer and a pharmaceutically acceptable carrier.
Pharmaceutically acceptable salts of the compounds of the formula (IA) are also contemplated.
It should be understood that the formula (IA) structurally encompasses all stereo isomers, including enatiomers and diastereomers, which may be contemplated from the chemical structure of the genus described herein.
Also contemplated are prodrugs of the compounds of the formula ( IA), including for example ester prodrugs.
Accordingly, one aspect of the present invention provides compounds of formula (IB):
Figure imgf000006_0001
Formula (IB)
or a pharmaceutically acceptable salt, prodrug or stereoisomer there of, wherein,
W] and W2 are independently can be selected from substituted or unsubstituted alkylene and preferably the substituent is selected from n-butyl, isobutyl or tertiary butyl;
Ria can be H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl ;
R2, R3, R2a and R3a are independently can be selected from H, C(0)R", C(0)OR", or S(0)2R";
R can be H, OH, halogen, NR", C(0)OR", C(0)NR" substituted or unsubstituted alkyl, substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl;
n can be an integer 1 -3;
R" can be H, substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted cycloalkyl, or substituted or unsubstituted heterocyclyl.
The present invention also provides the process for making the compounds of formula (EB).
The present invention further provides a pharmaceutical composition comprising a therapeutically effective amount of a compound or combination of compounds of the present invention, or a pharmaceutically acceptable salt form, stereoisomer and a pharmaceutically acceptable carrier.
Pharmaceutically acceptable salts of the compounds of the formula (EB) are also contemplated.
It should be understood that the formula (EB) structurally encompasses all stereo isomers, including enatiomers and diastereomers, which may be contemplated from the chemical structure of the genus described herein. Also contemplated are prodrugs of the compounds of the formula (IB), including ester prodrugs.
According to one embodiment, there is provided a compound of formula (IA), wherein W | is a bond.
According to one embodiment, there is provided a compound of formula
(IA), wherein W2 is a bond.
According to one embodiment, there is provided a compound of formula (IA), wherein W2 is a -CH(t-butyl)- and -CH(iso-butyl)-.
According to one embodiment, there is provided a compound of formula (IA), wherein R5 and R6 are independently methyl.
According to one embodiment, there is provided a compound of formula (IA), wherein Rsa and R6a are independently methyl.
According to one embodiment, there is provided a compound of formula (IA), wherein R4 , R7, Rg R7a and Rsa aje independently H.
According to one embodiment, there is provided a compound of formula
(IA), wherein R | is pyridine.
According to one embodiment, there is provided a compound of formula (IA), wherein R | is phenyl.
According to one embodiment, there is provided a compound of formula (IA), wherein R , is H.
According to one embodiment, there is provided a compound of formula (IA), wherein R ) is 2-(morpholin-4-yl)ethoxy.
According to one embodiment, there is provided a compound of formula (IA), wherein nx is 1 .
According to one embodiment, there is provided a compound of formula
(IA), wherein ny is 1 .
According to one embodiment, there is provided a compound of formula (IA), wherein ny is 0.
According to one embodiment, there is provided a compound of formula (IA), wherein R2, R3, R2a and R3a are independently H, -C(0)CH3, -C(0)OCH3, -
C(0)0-t-butyl, or -C(0)Bz.
According to one embodiment, there is provided a compound of formula (ΓΒ), wherein W | is methylene substituted by t-butyl and methylene substituted by iso-butyl . According to one embodiment, there is provided a compound of formula (EB), wherein W2 is methylene substituted by t-butyl.
According to one embodiment, there is provided a compound of formula (IB), wherein each R2, R3, R2a and R3a are independently H, and -C(0)OCH3.
According to one embodiment, there is provided a compound of formula
(IA), wherein R) a is morpholin-4-yl; 4-ethylpiperazin- l -yl ; and 1 ,2,3,4- tetrahydroquinolin- 1 -yl.
According to one embodiment, there is provided a compound of formula (EB), wherein R4 is H.
According to one embodiment, there is provided a compound of formula
(EB), wherein n is 2 and 3.
According to one embodiment, the representative compounds of fromula ( 1 ) which are illustrative in nature only and are not intended to limit to the scope of the invention (Nomenclature has been generated from Chem. Draw Ultra 1 1 .0 version):
( l S,3R)-3-acetamido-N-((2S,3S)-4-(2-(( l S,3R)-3-acetamido-2,2- dimethylcyclobutanecarbonyl)- l -(4-(pyridin-2-yl)benzyl)hydrazinyl)-3-hydroxy- l - phenylbutan-2-yl)-2,2-dimethylcyclobutanecarboxamide (Compound 1 ),
Methyl ( l R,3S)-3-((2S,3S)-4-(2-(( l S,3R)-3-methylcarbamato-2,2- dimethylcyclobutanecarbonyl)- l -(4-(pyridin-2-yl)benzyl)hydrazinyl)-3-hydroxy- l - phenylbutan-2-ylcarbamoyl)-2,2-dimethylcyclobutylcarbamate (Compound 2),
Teil-buty] ( l S,3R)-3-((2S,3S)-4-(2-(( l R,3S)-3-(tert- butyloxycarbonylamino)-2,2-dimethylcyclobutanecarbonyl)- l -(4-(pyridin-2- yl)benzyl)hydrazinyl )-3-hydroxy- l -phenylbutan-2-ylcarbamoyl)-2,2- dimethylcyclobutylcarbamate (Compound 3),
Benzyl ( l S,3R)-3-((2S,3S)-4-(2-(( l R,3S)-3-(benzyloxycarbonylamino)-2,2- dimethylcyclobutanecarbonyl)- 1 -(4-(pyridin-2-yl)benzyl)hydrazinyl)-3-hydroxy- 1 - phenylbutan-2-ylcarbamoyl)-2,2-dimethylcyclobutylcarbamate (Compound 4),
Methyl ( l S,3R)-3-((2S,3S)-3-hydroxy-4-(2-(( l R,3S)-3- (methoxycarbonylamino)-2,2-dimethylcyclobutanecarbonyl)- l -(4-(pyridin-2- yl)benzyl)hydrazinyl)- 1 -phenylbutan-2-ylcarbamoyl)-2,2- dimethylcyclobutylcarbamate (Compound 5),
Methyl ( 1 R,3S)-3-((2S,3S)-3-hydroxy-4-(2-((S)-2-(methoxycarbonylamino)- 3,3-dimethylbutanoyl)- l -(4-(pyridin-2-yl)benzyl)hydrazinyl)- l -phenylbutan-2- ylcarbamoyl)-2,2-dimethylcyclobutylcarbamate (Compound 6), Methyl (S)-l-(2-((2S,3S)-3-((lS,3 )-3-(tert-butoxycarbonylamino)-2,2- dimethylcyclobutanecarboxamjdo)-2-hydroxy-4-phenylbutyl)-2-(4-(pyridin-2- yl)benzyl)hydrazinyl)-3,3-dimethyl- 1 -oxobutan-2-ylcarbamate (Compound 7),
Methyl (S)-l-(2-((2S,3S)-3-((lS,3R)-3-acetamido-2,2- dimethylcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-(4-(pyridin-2- yl)benzyl)hydrazinyl)-3,3-dimethyl- 1 -oxobutan-2-ylcarbamate (Compound 8),
Methyl (S)-l-(2-((2S,3S)-2-hydroxy-3-((lR,3S)-3-(methoxycarbonylamino)-
2.2- dimethylcyclobutanecarboxamido)-4-phenylbutyl)-2-(4-(pyridin-2- yl)benzyl)hydrazinyl)-3,3-dimethyl- 1 -oxobutan-2-ylcarbamate (Compound 9),
Methyl (S)- 1 -(2-((2S,3S)-3-(( 1 R,3S)-3-(tert-butoxycarbonylamino)-2,2- dimethylcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-(4-(pyridin-2- yl)benzyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate (Compound 10),
Methyl (S)-l-(2-((2S,3S)-3-((lR,3S)-3-(benzyloxycarbonylamino)-2,2- dimethylcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-(4-(pyridin-2- yl)benzyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate (Compound 11),
Methyl (S)-l-(2-benzyl-2-((2S,3S)-2-hydroxy-3-((lS,3R)-3- (methoxycarbonylamino)-2,2-dimethylcyclobutanecarboxamido)-4- phenylbutyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate (Compound 12),
Methyl (S)-l-(2-((2S,3S)-3-((lS.3R)-3-acetamido-2,2- dimethylcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-benzylhydrazinyl)-
3.3- dimethyl-l-oxobutan-2-ylcarbamate (Compound 13),
Methyl (S)-l-(2-benzyl-2-((2S,3S)-3-((lS,3R)-3-(tert- butoxycarbonylamino)-2,2-dimethylcyclobutanecarboxamido)-2-hydroxy-4- phenylbutyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate (Compound 14), Methyl (S)-l-(2-((2S,3S)-3-((lS,3R)-3-acetamido-2,2- dimethylcyclobutanecaiboxamido)-2-hydroxy-4-phenylbutyl)-2-(biphenyl-4- ylmethyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate (Compound 15),
Methyl (S)-l-(2-(biphenyl-4-ylmethyl)-2-((2S,3S)-2-hydroxy-3-((lS,3R)-3- (methoxycarbonylamino)-2,2-dimethylcyclobutanecarboxamido)-4- phenylbutyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate (Compound 16),
Methyl (S)-l-(2-(biphenyl-4-ylmethyl)-2-((2S,3S)-3-((lS,3R)-3-(tert- butoxycarbonylamino)-2,2-dimethylcyclobutanecarboxamido)-2-hydroxy-4- phenylbutyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate (Compound 17), Methyl (S)- 1 -(2-(biphenyl-4-ylmethyl)-2-((2S,3S)-2-hydroxy-3-(( 1 R,3S)-3- (methoxycarbonylamino)-2,2-dimethylcyclobutanecarboxamido)-4- phenylbutyl)hydrazinyl)-3,3-dimethyl- 1 -oxobutan-2-ylcarbamate (Compound 18),
Methyl (S)-l-(2-(biphenyl-4-ylmethyl)-2-((2S,3S)-3-((lR,3S)-3-(tert- butoxycarbonylamino)-2,2-dimethylcyclobutanecarboxamido)-2-hydroxy-4- phenylbutyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate (Compound 19),
Methyl (S)-l-(2-((2S,3S)-3-((lR,3S)-3-(benzyloxycarbonylamino)-2,2- dimethylcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-(biphenyl-4- ylmethyl)hydrazinyl)-3,3-dimethyl- 1 -oxobutan-2-ylcarbamate (Compound 20), Methyl (S)-l-(2-((2S,3S)-2-hydroxy-3-((lS,3R)-3-(methoxycarbonylamino)-
2,2-dimethylcyclobutanecarboxamido)-4-phenylbutyl)-2-(4-(pyridin-2- yl)benzyl)hydrazinyl)-4-methyl- 1 -oxopentan-2-ylcarbamate (Compound 21 ),
Methyl (S)- 1 -(2-((2S,3S)-3-(( 1 S,3R)-3-(tert-butoxycarbonylamino)-2,2- dimethylcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-(4-(pyridin-2- yl)benzyl)hydrazinyl)-4-methyl-l-oxopentan-2-ylcarbamate (Compound 22),
Methyl (S)-l-(2-((2S.3S)-3-((lS,3R)-3-acetamido-2,2- dimethylcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-(4-(pyridin-2- yl)benzyl)hydrazinyl)-4-methyl- 1 -oxopentan-2-ylcarbamate (Compound 23),
Methyl (S)-l-(2-((2S,3S)-2-hydroxy-3-((lR,3S)-3-(methoxycarbonylamino)- 2,2-dimethylcyclobutanecarboxamido)-4-phenylbuty )-2-(4-(pyridin-2- y])benzyl)hydrazinyl)-4-methyl- 1 -oxopentan-2-ylcarbamate (Compound 24),
Methyl (S)-l-(2-((2S,3S)-3-((lR,3S)-3-(tert-butoxycarbonylamino)-2,2- dimethylcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-(4-(pyridin-2- yl)benzyl)hydrazinyl)-4-methyl- 1 -oxopentan-2-ylcarbamate (Compound 25),
Methyl (S)-l-(2-((2S,3S)-3-((lR,3S)-3-(benzyloxycarbonylamino)-2,2- dimethylcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-(4-(pyridin-2- yl)benzyl)hydrazinyl)-4-methyl-l-oxopentan-2-ylcarbamate (Compound 26),
Methyl N-[(lS)-l-{N'-[(2S,3S)-2-hydroxy-3-[(2S)-2- [(methoxycarbonyl)amino]-3,3-dimethylbutanamido]-4-phenylbutyl]-N'-({4-[2- (mo holin-4-yl)ethoxy]phenyl}methy])hydrazinecarbonyl }-2,2- dimethylpropyl]carbamate (Compound 27),
Methyl N-[(lR)-l-{[(lS,2R)-l-hydroxy-l-[(2S)-2- [(methoxycarbonyl)amino]-3,3-dimethyl-N,-({4-[2-(mo holin-4- yl)ethoxy]phenyl}methyl)butanehydrazido]-3-phenylpropan-2-yl]carbamoyl}-3- methylbutyl]carbamate (Compound 28),
Methyl N-[(lS)-l-{N'-[(lS,2R)-l-hydroxy-2-{[(lS,3R)-3- [(methoxycarbonyl)amino]-2,2-dimethylcyclobutyl]formamido)-3-phenylpropyl]- N'-( { 4-[2-(mo holin-4-yl)ethoxy]phenyl } methyl)hydrazinecarbonyl } -2,2- dimethylpropyl]carbamate (Compound 29),
Methyl N-[(lS)-l-{N'-benzyl-N'-[(lS,2R)-2-{[(lS,3R)-3-{[(tert- butoxy)carbonyl]amino } -2,2-dimethylcyclobutyl]formamido } - 1 -hydroxy-3- phenylpropyl]hydrazinecarbonyl } -2,2-dimethylpropyl]carbamate (Compound 30), Methyl N-[(lR,3S)-3-{[(lS,2R)-l-hydroxy-l-{ l-[(lR,3S)-3-
[(methoxycarbonyl)amino]-2,2-dimethylcyclobutyl]-N'-{ [4-(pyridin-2- yl)phenyl]methyl } formohydrazido } -3-phenylpropan-2-yl]carbamoyl } -2,2- dimethylcyclobutyl]carbamate (Compound 31),
Methyl N-[(lS)-l-(N'-[(lS,2R)-2-{ [( lS,3R)-3-{ [(tert- butoxy)carbonyl]amino}-2,2-dimethy]cyclobutyl]formamido}-l-hydroxy-3- phenylpropyl]-N'-({4-[2-(mo holin-4- yl)ethoxy]phenyl } methyljhydrazinecarbonyl } -2,2-dimethylpropyl]carbamate (Compound 32),
Methyl N-[(lS)-l-(N*-[(lS,2R)-2-{ [(lR,3S)-3-( [(tert- butoxy)carbonyl]amino } -2,2-dimethylcyclobutyl]formamido } - 1 -hydroxy-3- phenylpropyl]-N,-({4-[2-(mo holin-4- yl)ethoxy]phenyl } methyl)hydrazinecarbonyl } -2,2-dimethylpropyl]carbamate (Compound 33),
Methyl N-[(lS)-l-{N'-[(lS,2R)-l-hydroxy-2-{ [(lS,3R)-3- [(methoxycarbonyl)amino]-2,2-dimethylcyclobutyl]formamido}-3-phenylpiOpyl]- N'-( { 4-[2-(mo holin-4-yl)ethoxy]phenyl } methyl)hydrazinecarbonyl } -2,2- dimethylpiopyl]carbamate (Compound 34),
Methyl N-[(lR)-l-{ [(lS,2R)-l-hydroxy-l-[(2S)-2- [(methoxycarbonyl)amino]-3,3-dimethyl-N'-({4-[2-(l,2,3,4-tetrahydroquinolin-l- yl)ethoxy]phenyl }methyl)butanehydrazido]-3-phenylpropan-2-yl]carbamoyl }-2,2- dimethylpropyl]carbamate (Compound 35),
Methyl N-[(lR)-l-{[(lS,2R)-l-[(2S)-N'-({4-[2-(4-ethylpiperazin-l- yl)ethoxy]phenyl }methyl)-2-[(methoxycarbonyl)amino]-3,3- dimethylbutanehydrazido]- 1 -hydroxy-3-phenylpropan-2-yl]carbamoyl } -2,2- dimethylpropyl]carbamate (Compound 36),
Methyl N-[( 1 R)- 1 - { [( 1 S,2R)- 1 -hydroxy- 1 -[(2S)-2-
[(methoxycarbonyl)amino]-3,3-dimethyl-N'-( { 4-[3-( 1 ,2,3,4-tetrahydroquinolin- 1 - yl)propoxy]phenyl } methyl)butanehydrazido]-3-phenylpropan-2-yl]carbamoyl } -2,2- dimethylpropyl]carbamate (Compound 37),
Methyl N-[( l S)- l - { N'-[(3S)-3-[(2S)-2- { [(tert-butoxy)carbonyl]amino } -4- methylpentanamido]-2-hydlΌxy-4-phenylbutyl]-N'-( { 4-[2-(mo holin-4- yl)ethoxy]phenyl } methyl)hydrazinecarbonyl } -2,2-dimethylpropyl]carbamate (Compound 38), or
pharmaceuticall y acceptable salts, solvates, stereo isomers, including prodrugs of compounds are also contemplated.
The compounds and pharmaceutical compositions described herein are useful in the treatment of diseases, conditions and/or disorders mediated by viral infections more particularly for treating, for example, HIV infection .
The present patent application further provides a method of treating a disease, condition and/or disorder mediated by viral infections more particularly HIV infection in a subject in need thereof by administering to the subject one or more compounds described herein in the amount effective to treat that infection.
Detailed Description of the Invention
The present invention relates to novel aza-peptides containing 2 ,2- disubstituted cyclobutyl and/or substituted alkoxy benzyl derivatives and, a composition for inhibiting Human Immunodeficiency Virus (HIV) and process for making the compounds.
The following defi nitions 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 solel y of carbon and hydrogen atoms, containi ng no unsaturation , having from one to eight carbon atoms, and which is attached to the rest of the molecule by a si ngle bond, e.g.. methyl, ethyl , n-propyl , 1 -methylethyl (isopropyl), n-butyl , n- pentyl , and 1 , 1 -dimethylethyl (t-butyl).
The term "alkenyl" refers to an aliphatic hydrocarbon group containing a carbon-carbon double bond and which may be a straight or branched chain having from 2 to about 10 carbon atoms, e.g., ethenyl , 1 -propenyl, 2-propenyl (allyl), iso- propenyl, 2-methyl- 1 -propenyl, 1 -butenyl, and 2-butenyl .
The term "alkylene" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation, having from one to five carbon atoms, and which is attached to the rest of the molecule by two sides bond, e.g., methylene, ethylene, n-propylene, - C(methyl)(isopropyl)-, -CH(isopropyl )-, -CH(n-biityl)-, -CH(t-butyl)-, -CH(iso- butyl)-, -CH2-CH(n-butyl)-, -CH2-CH(iso-butyl )-, or -CH2-CH(t-butyl)-.
The term "acyl group" is used to denote a linear or branched aliphatic acyl group (preferably a C2.6 alkanoyl group) or an aromatic acyl group, which contains 2 to 10 carbon atoms. Examples include an acetyl group, a propionyl group, a pivaloyl group, a butyryl group, an isobutyryl group, a valeryl group and a benzoyl group, with an acetyl group being preferred.
The term "alkoxy group" is used to denote a linear or branched alkoxy group containing 1 to 6 carbon atoms. Preferred are C | .4 alkoxy groups including a methoxy group, an ethoxy group, a propoxy group, an isopropoxy group, an n- butoxy group, an isobutoxy group and a tert-butoxy group.
The term "alkoxycarbonyl group" isused to denote a structure composed of a linear or branched C | .5 alkoxy group and a carbonyl group. Preferred are C2-5 alkoxycarbonyl groups including a methoxycarbonyl group, an ethoxycarbonyl group, a propoxycarbonyl group, an isopropoxycarbonyl group and a butoxycarbonyl group. Among them, a methoxycarbonyl group is preferred.
The term "cycloalkyl" denotes a non-aromatic mono or multicyclic ring system of from 3 to about 1 2 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 "cycloalkylalkyl" refers to a cyclic ring-contai ning radical having from 3 to about 8 carbon atoms directly attached to an alkyl group. The cycloalkylalkyl group may be attached to the main structure at any carbon atom in the alkyl group that results in the creation of a stable structure. Non-limiting examples of such groups include cyclopropyl methyl, cyclobiitylethyl , and cyclopentylethyl . 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.
"Substituted" refers to 1 -3 substituents on the same position or on different positions with the same groups or different groups.
" Alkynyl " refers to alkynyl groups having from 2 to 6 carbon atoms and having at least one alkynyl saturation, example for such group includes acetylenyl, propargyl.
"Carbonyloxy" refers to a group such as -C(0)0.
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; and the ring radical may be partially or fully saturated (i.e., heterocyclic or heteroaryl). Examples of such heterocyclic ring radicals include, but are not limited to, azetidinyl , acridinyl, carbazolyl, cinnolinyl, indolizinyl, naphthyridinyl, perhydroazepinyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pyridyl, pteridinyl, purinyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrazoyl, imidazolyl, tetrahydroisouinolyl, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2- oxopyrrolidinyl, 2-oxoazepinyl, azepinyl, pyrrolyl, 4-piperidonyl, pyrrolidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolinyl, oxazolidinyl, triazolyl, isoxazolyl, isoxasolidinyl , morpholinyl, thiazolyl, thiazolinyl, thiazolidinyl, isothiazolyl, quinuclidinyl , isothiazolidinyl, indolyl, isoindolyl, indolinyl, isoindolinyl , octahydroindolyl, octahydroisoindolyl, quinolyl, isoquinolyl, decahydroisoquinolyl, benzimidazolyl, thiadiazolyl. benzothiazolyl, benzooxazolyl, furyl, tetrahydrofuryl , tetrahydropyranyl, thienyl, benzothienyl, thiamorpholinyl, thiamorpholiny] sulfoxide, thiamorpholinyl sulfone, dioxaphospholanyl, oxadiazolyl . The heterocyclic ring radical may be attached to the main structure at any heteroatom or carbon atom that results in the creation of a stable structure. The term "heteroaryl" refers to an aromatic heterocyclic ring radical. The heteroaryl ring radical may be attached to the main structure at any heteroatom or carbon atom that results in the creation of a stable structure.
The term "heteroarylalkyl" refers to a heteroaryl ring radical directly bonded to an alkyl group. The heteroarylalkyl 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.
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 alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkenylalkyl, 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, substituted or unsubstiuted guanidine, -COORx, -C(0)R\ -C(S)R\ -C(0)NRxRy, -C(0)ONRxRy, -NRxCONRyRz, -N(Rx)SORy, -N(Rx)S02Ry, -(=N- N(Rx)Ry), -NRxC(0)ORy, -NRxRy, -NRxC(0)Ry, -NRxC(S)Ry, -NRxC(S)NRyR\ - SONR Rv, -S02NRxRy, -OR\ -ORxC(0)NRyRz, -OR C(0)ORy, -OC(0)Rx, -
OC(0)NRxRy, -R NRvC(0)Rz, -RxORy, -RxC(0)ORv, -RxC(0)NRyRz, -RxC(0)Ry, - RxOC(0)Ry, -SRX, -SORx, -S02Rx, and -ON02, wherein R\ Ry and Rz are independently selected from hydrogen, substituted or unsubstituted alkyl, haloalkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, 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 heterocyclylalkyl ring, substituted or unsubstituted heteroarylalkyl, or substituted or unsubstituted heterocyclic ring. The substituents in the aforementioned "substituted" groups cannot be further substituted. For example, when the substituent on "substituted alkyl" is "substituted aryl", the substituent on "substituted aryl" cannot be "substituted alkenyl". The term "prodrug" means a compound that is transformed in vivo to yield a compound of Formula (I) (IA) or (ΓΒ) or a pharmaceutically acceptable salt, hydrate or solvate 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).
A "therapeutically effective amount" means, for example, 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 compound of the invention may form salts. Non-limiting examples of pharmaceutically acceptable salts forming part of the invention include salts derived from inorganic/organic acids or bases and amino acids salts. Certain compounds of the invention are capable of existing in stereoisomeric forms (e.g. diastereomers and enantiomers). With respect to the overall compounds described by the Formula (I), Formula (LA) or Formula (LB), the invention extends to these stereoisomeric forms and to mixtures thereof. To the extent prior art teaches synthesis or separation of particular stereoisomers, the different stereoisomeric forms of the invention may be separated from one another by the method known in the art, or a given isomer may be obtained by stereospecific or asymmetric synthesis. Tautomeric forms and mixtures of compounds described herein are also contemplated.
Pharmaceutical Compositions
The pharmaceutical compositions provided in the invention include at least one compound described herein and at least one pharmaceutically acceptable excipient (such as a pharmaceutically acceptable carrier or diluent). Preferably, 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 bei ngs. The compound of the present invention may be associated with a pharmaceuticall y acceptable excipient (such as a carrier or a diluent) or be diluted by a carrier.
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, pecti n, 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 auxil iary agents, wetting agents, emulsifyi ng 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 i ngredient after administration to the subject by employing procedures known in the a . The pharmaceutical compositions described herein may be prepared, e.g., as described in Remington: The Science and Practice of Pharmacy, 20lh Ed., 2003 (Lippincott Williams & Wilkins).
The pharmaceutical compositions may be, for example, capsules, tablets, aerosols, solutions, suspensions 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 preferred.
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. Preferable 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 tabletting techniques may contain: ( 1 ) Core: Active compound (as free compound or salt thereof), colloidal silicon dioxide (Aerosil®), microcrystalline cellulose (Avicel®), modified cellulose gum (Ac-Di-Sol®), and magnesium stearate; (2) Coating: HPMC, Mywacett 9-40 T and acylated monoglyceride.
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, preferably 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* 103 cells /well) will be incubated 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 1 * 106 Sup-T l 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 HIV 1 93 |N t o i 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.
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 chemothera, 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, 191 1 - 15( 1986); Pennington etal., 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 assy 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, 5306-53 1 1 ; Popik et al., Journal of virology, 2002, 76, 4709-4722; Harrigton et al., Journal of Virology Methods, 2000, 88, 1 1 1 - 1 15; Roos et al., Virology 2000, 273, 307-3 15; 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. EP0989862 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 patent application further provides a method of treating a disease, condition and/or disorder mediated by viral infections in a subject in need thereof by administering to the subject a therapeutically effective amount of a compound or a pharmaceutical composition of the present invention.
Diseases, conditions, and/or disorders that are mediated by viral infections are believed to include, but are not limited to, HIV infection, HBV, HCV, a retroviral infection genetically related to HIV, AIDS, or respiratory disorders (including adult respiratory distress syndrome (ARDS).
The compounds of the present invention can also obtain synergistic effects in the prevention or treatment of the above diseases when used suitably in combination with existing drugs. The administered dose may be decreased in comparison with administration of either drug alone, and in addtion adverse effects of co- administrated drugs can be avoided or reduced.
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 4. 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 stereo isomers of the compounds in these schemes, unless otherwise specified, are also encompassed within the scope of this invention.
Figure imgf000021_0001
The compounds of Formulae 6 and 13 can be prepared by the above procedure as described in Scheme 1 by adopting the methods published in Tetrahedron :Assymmetry 19(2008302-308) (wherein, P is a protecting group (for example alkyl or aralkylyl protecting groups such as, methyl, ethyl, propyl, isopropyl, benzyl, tertiorybutyl, benzyl carbonyl (Cbz) or tertiorybutyloxy carbonyl
(Boc)), Ra and Rb are independently can be selected from H, C(0)R", C(0)OR", or S(0)2R"). Commercially available ve benone (formula 1 ) was oxidized with an oxidizing agent like ruthenium chloride/sodium periodate in a solvent system like CC14-ACN-H20 to give the carboxylic acid of compounds of formula 2. The carboxylic acid of compounds of formula 2 is protected with protecting agent to give the acid protected of compounds of formula 3 in the presence of a base like cesium carbonate, or in the presence of a dehydrating agent like DCC or an acid catalyst or the like in the solvents such as DMF, or the like. The acid protected compounds of formula 3 are subjected to Schmidt rearrangement by treatment with sodium azide and methane sulphonic acid or the like to give the N-acyl compounds of formula 4. The N-acyl of compounds of formula 4 was Hydrogenolysed (when P is Bn) in the presence of Pd-C EtOAc or the like to give the N-acyl amino acid of compounds of formula 5. The N-acyl amino acid of compounds of formula 5 can be acid hydrolised in the presence of hydrogen chloride or the like and treated with BOC anhydride in appropriate conditions to give corresponding carbamate more particularly the tertiary butyl carbamate of compounds of formula 6.
Alternatively, the carboxylic acid of compounds of formula 2 can be converted in to the azide compounds of formula 7 (through a Curtius rearrangement) in the presence of chloroformates such as for example, ethyl chloroformate or the like and metal azides such as for example, sodium azide or the like in the solvents such as for example, acetone, dichloromethane, dimethylformamide or the like. The azide compounds of formula 7 can be converted to the N-protected compounds of formula 8 as described in J. Org. Chem. 1987, 52, 689 in the presence of alcohols such as for example, t-butyl alcohol, benzyl alcohol or the like in the solvents such as, for example toluene, acetone, dichloromethane, dimethylformamide or the like. The N-protected compounds of formula 8 can be converted to the acid compounds of formula 9 by Lieben degradation of methyl ketone function of formula 8 in the presence of dioxane and aqueous sodiumhypobromite or the like. The acid compounds of formula 9 can be converted to the corresponding ester compounds of formula 10 by alkylation of resultant acid in the presence of alkyl halide such as for example, methyl iodide, ethyl iodide or the like and a base such as potassium or cesium carbonate in a solvent like dimethyl formamide.. The alkyl ester compounds of formula 10 can be N- deprotected in the presence of deprotecting agents such as such as for example, palladium on carbon or the like to give the deprotected compounds of formula 1 1 (also Compounds of formula 1 1 can be prepared either from alpha-pinene or verbenone. Alpha-pinene can be converted into verbenone by allylic oxidation). The deprotected compounds of formula 1 1 can be converted to the N-substituted compounds of formula 12 with corresponding chloroformates or the like in the solvents such as, for example, dichloromethane, dimethylformamide and a base like triethyl amine or the like. The N-substituted compounds of formula 12 can be hydrolysed in the presence of base or hydrogenolysed in the presence of Pd- C EtOAc when ester is a benzyl ester or the like to give the acid compounds of formula 13.
Figure imgf000023_0001
The compounds of Formula 23 can be prepared by the above procedure as described in Scheme 2 (wherein, P is a protecting group (for example alkyl or aralkylyl protecting groups such as, methyl, ethyl, propyl, isopropyl, benzyl or tertiorybutyl, benzyl carbonyl (Cbz) or tertiorybutyloxy carbonyl (Boc) and the remaining R-groups are same as defined above) by Schotten-Baumann acylation. The amino carboxylic acid compounds of formula 14 can be reacted with corresponding substituents for example, chloroformates or the like in the suitable conditions such as, for example, dioxane, sodium hydroxide or the like to give the substituted amino compounds of formula 15. The substituted amino compounds of formula 15 can be reacted with hydrazinecarboxylates to give the protected hydrazine compounds of formula 16 in the presence of coupling agents such as 1 - Ethyl-3-(3-Dimethylaminopropyl)carbodiimide (EDCI), hydroxybenzotriazole (HOBt) or the like in the solvents such as for example, N-methyl morpholine (NMM), dimethylformamide, diiospropyl ethylamine, dichloromethane, ethyl acetate or the like. The protected hydrazine compounds of formula 16 can be deprotected in suitable conditions such as, for example HCl in dioxane, Hal or the like to give the deprotected hydrazine compounds of formula 17. The deprotected hydrazine compounds of formula 17 can be reacted with substituted or unsubstituted benzaldehyde compounds of formula 18 to give the corresponding (benzylidene)hydrazinyl compounds of formula 19 in the presence of alcohols such as, for example, isopropanol, ethanol, methanol or the like. The (benzylidene)hydrazinyl compounds of formula 19 can be hydrogenolysed in the presence of hydrogenolysing agents such as for example, sodiumcyanoborohydride/p-tolunesulfonicacid, palladium on carbon, sodiumformate or the like in the solvents such as ethanol, water or the like to give the corresponding benzyl hydrazine compounds of formula 20. The benzyl hydrazine compounds of formula 20 can be reacted with N-protected amino epoxide compounds of formula 21 to give the corresponding N-protected peptide compounds of formula 22 in the presence of solvents such as for example, isopropanol, ethanol, ethyl acetate or the like. The N-protected peptide compounds of formula 22 can be deprotected by hydrogen chloride in solvents such as dioxane or tetrahydrofuran or the like to give the free amino peptide compounds of formula 23.
Figure imgf000024_0001
The compounds of Formula 27 [Formula (I), wherein W | and W2 are a bond; Z| and Z2 are 2,2-dimethylcyclobutyl; R2 and R2a are Ra and R3 and R a are Rb', and the remaining R-groups are same as defined above; each P| and P2 are protecting groups (for example alkyl or aralkylyl protecting groups such as, methyl, ethyl, propyl, isopropyl, benzyl or tertiorybutyl, benzyl carbonyl (Cbz) or tertiorybutyloxy carbonyl (Boc)] can be prepared by the above procedure as described in Scheme 3. The N-protected benzyl hydrazine compounds of formula 24 can be reacted with N- protected amino epoxide compounds of formula 21 (as described in Bio org. Med. Chem. Lett.1993, 3, 2837-2842) to give the corresponding N-protected peptide compounds of formula 25 in the presence of solvents such as for example, isopropanol, ethanol, ethyl acetate or the like. The N-protected peptide compounds of formula 25 can be deprotected by hydrogen chloride in the solvents such as
5 tetrahydrofuran or the like to give the free amino peptide compounds of formula 26.
Finally, the free amino peptide compounds of formula 26 can be reacted with the acid compounds of formula 13 to give the final compounds of formula 27 [Formula (I), wherein W | and W2 are a bond; Z| and Z2 are 2,2-dimethylcyclobutyl; R2 and R2a are Ra and R3 and R a are Rb] in the presence of coupling agents such as
10 N,N,N',N'-tetramethyl-o-(benzotriazol- l -yl)uronium tetrafluoroborate (TBTU), 1 -
Ethyl-3-(3-Dimethylaminopropyl)carbodiimide (EDCI), hydroxybenzotriazole (HOBt) or the like in the solvents such as for example, N-methyl morpholine (NMM), dimethylformamide, diiospropyl ethylamine, dichloromethane, ethyl acetate or the like.
Figure imgf000025_0001
J 5
The compounds of Formula (I) (wherein W | , W2, Z| , Z2 and each R-group are same as defined above) can be prepared by the above procedure as described in Scheme 4. The free amino peptide compounds of formula 23 can be reacted with the acid compounds of formula 13 (which was obtained in scheme- 1 ) to give the final
20 compounds of formula 27 (which was obtained in scheme-2) in the presence of coupling agents such as N,N,N',N'-tetramethyl-o-(benzotriazol- l -yl)uronium tetrafluoroborate (TBTU), 1 -Ethyl-3-(3-Dimethylaminopropyl)carbodiimide (EDCI). hydroxybenzotriazole (HOBt) or the like in the solvents such as for example, N-methyl morpholine (NMM), dimethylformamide, diiospropyl
25 ethylamine, dichloromethane, ethyl acetate or the like. 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 : Synthesis of ( l S,3R)-3-acetamido-2,2- dimethylcvclobutanecarboxylic acid:
Figure imgf000026_0001
Step 1: synthesis of ( lS,3R)-3-acetyl-2,2-dimethylcyclobutanecarbo.xylic acid:
Figure imgf000026_0002
A stirred solution of Verbenone (about 10 g, 66.57 mmol) in CC14: ACN: H20 ( 140 ml) was added NaI04 (about 56.9 g, 266.3 mmol) followed by RuCl3.H20
(catalytic) was added at 0°C and stirred at room temperature for about 12 hours. After completion of the reaction (monitored by TLC), reaction mixture was diluted with EtOAc, filtered through celite bed and the organic layer was separated then the aqueous layer was extracted with EtOAc ( 100 ml x 2). Combined organic layers were dried with Na2S04 and concentrated under reduced pressure. The resulting crude was stirred in hexane, forming the solid. Hexane was decanted, and the resulting solid was dried under vacuum. Wt: 8.56 g: Yield: 76.0 %; Ή NMR (300 MHz, CDC13): δ 2.91 -2.77 (m, 2H), 2.63 (q, 1 H, J = 9Hz), 2.07 (s, 3H), 1.86- 1.95 (m, 1 H), 1 .40 (s, 3H), 0.97 (s, 3H); Mass: [M+ l ]+ 17 1 ( 10%), [M+Na]+ 193 (72%); IR ( Br, cm" 1 ): 3218, 1737, 1694, 1467, 1368, 1283, 1 174, 1080, 836, 703 ; M.R:
109.5 °C - 120.7 °C.
Step 2; Synthesis of (lS, 3R)-benzyl 3-acetyl-2,2-dimethylcyclobutanecarboxylate:
Figure imgf000027_0001
A stirred solution of (lS,3R)-3-acetyl-2,2-dimethylcyclobutanecarboxylic acid (Step 1, about 6.0 g, 35.29 mmol) in DMF (20 ml) Cs2C03 (13.8 g, 42.34 mmol) and BnBr (5.06 ml, 42.34 mmol) were added slowly portion wise at about 0 °C, after addition was completed the reaction was stirred at room temperature for about 2 hours. After completion of the reaction (monitored by TLC) the reaction mixture was diluted with EtOAc (100 ml) and washed with water (50 ml), brine (50 ml) and dried with Na2S04, the solvent was evaporated and purified by silica gel column (60-120, elution 4% EtOAc in hexane) chromatography to afford the title compound as a pale yellow syrup. Wt: 5.6 g: Yield: 61.5 %; Ή NMR (300 MHz, CDCI3): δ 7.42-7.31 (m, 5H), 5.13 (s, 2H), 2.93-2.81 (m, 2H), 2.68 (q, 1H, J = 10.5 Hz,), 2.05 (s, 3H), 1.97-1.88 (m, 1H), 1.43 (s, 3H), 0.87 (s, 3H); Mass: [M+l]+ 261(20%), [M+Na]+283 (87%); IR (KBr, cm"1): 2889, 1732, 1708, 1498, 1456, 1329, 1279, 1183, 1027, 954, 752, 698.
Stpe 3: synthesis of ( lS,3R)-benzyl 3-acetamido-2,2- dimethylcyclob tanecarboxylat
Figure imgf000027_0002
To a stirred solution of (lS,3R)-benzyl 3-acetyl-2,2- dimethylcyclobutanecarboxylate (Step 2, about 1.5 g, 5.76 mmol) in monoglyme (about 18.75 ml) at about -45 °C, NaN3 (1.12 g, 17.30 mmol) and MeSOjH (10.5 ml) were added drop wise slowly and the reaction was allowed to stir at room temperature for about 48 hours. After completion of the reaction (monitored by TLC), the reaction mixture was neutralized with aqueous ammonia solution, solvent was evaporated, diluted with DCM and washed with water, brine dried with Na2S04 then the solvent was evaporated under reduced pressure. The resulting crude purified by silica gel column (60/120mesh, elution: 30% EtOAc in Hexane) chromatography to afford the title compound as off white solid. Wt: 1.3 g; Yield: 82.2 %; Ή NMR (300 MHz, CD3OD): 57.42-7.31 (m, 5H), 5.59 (d, 1H, J = 5.7 Hz), 5.12 (s, 2H), 4.13 (q, 1H.7 = 9.3 Hz), 2.69-2.63 (m, 1H), 2.38-2.34 (m, 1H), 2.13-2.06 (m, 1H), 2.05 (s, 3H), 1.30
(s, 3H), 0.86 (s, 3H); Mass: [M+l]+276 (100%), [M+Na]+298 (66%); IR (KBr, cm 1): 3270, 2963, 2866, 1728, 1647, 1571 , 1455, 1303, 1 151 , 107 1 , 1019, 961 , 755, 742; M.R: 85.6 °C -97.5 °C.
Step 4: Synthesis of (lS,3R)-3-acetamido-2,2-dimethylcyclobutanecarbox}iic acid:
A stirred solution of ( l S,3R)-benzyl 3-acetamido-2,2- dimethylcyclobutanecarboxylate (Step 3, about 0.650 g, 2.36 mmol) in EtOAc ( 10 ml) 10 % Pd/C (catalytic amount) was added at room temperature and the reaction mixture was stirred under H2 gas atmosphere at room temperature for about 1 hour. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through celite bed, the obtained filtrate concentrated under reduced pressure to afford title compound as an off white solid. Wt: 0.240 g: Yield: 54.9%; Ή NMR (300 MHz, CD3OD): δ 8.09 (d, J = 3.0 Hz, 1 H), 4.08-3.96 (m, 1 H), 2.62-2.58 (m, 1 H), 2.20 -2. 15 (m, 2H), 1.93 (s, 3H), 1.26 (s, 3H), 0.92 (s, 3H); Mass: [M+ l ]+ 186 (70%), [M+Na]+ 208 (93%); IR (KBr, cm 1 ): 3349, 2962, 1697, 1624, 1557, 1428, 1377, 1257, 1075. 988, 920, 736; M.R: 194.7 °C -216.2 °C.
Intermediate 2: Synthesis of ( l S,3R)-3-(methoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid:
Figure imgf000028_0001
A stirred suspension of ( l S,3R)-3-acetamido-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 1 , about 2.2 g, 1 1 .89 mmol) in 3N HCl (35 ml) was refluxed for about 2 hours. After completion of the reaction (monitored by TLC), the solvent was evaporated and basified (PH = 8-9) with 2N NaOH, at 0 °C dioxane (5 ml), followed by methyl chloroformate (2. 15 ml, 27.93 mmol) was added then the reaction was allowed to stir at room temperature for about
16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with I N HCl and extracted with EtOAc, washed with water, brine and dried with Na:S04 then the organic layer concentrated under reduced pressure. The resulting crude was purified by silica gel column ( 100-200 mesh, elution: 30% EtOAc in Hexane) chromatography to afford the title compound as an off white solid. Wt: 1 .4 g: Yield: 50%; Ή NMR (300 MHz, CD3OD): δ 7. 10 (d, 1 H, J = 7.2 Hz), 3.80-3.77 (m, 1 H), 3.61 (s, 3H), 2.60-2.54 (m, 1 H), 2.21 (m, 2H), 1 .25 (s, 3H), 0.92 (s, 3H); Mass: [M-l]"200 (10%); IR (KBr, cm"1): 3371, 3322, 2874, 1702, 1554, 1464, 1259, 1197, 1057, 932, 813, 756; M.R: 116.9 °C -122.7 °C.
Intermediate 3: Synthesis of (lS,3R)-3-(tert-butoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid:
Figure imgf000029_0001
A stirred suspension of (lS,3R)-3-acetamido-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 1, about 0.5 g, 2.702 mmol) in 3N HC1 (8 ml) was refluxed for about 2 hours. After completion of the reaction (monitored by TLC), the solvent was evaporated and basified (PH = 8-9) with 2N
NaOH at about 0°C then dioxane (3 ml) followed by (Boc)20 (about 1.24 ml, 5.405 mmol) were added and the reaction was allowed to stir at room temperature for about 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with aqueous saturated citric acid solution and extracted with EtOAc, the organic layer was washed with water, brine, dried with Na2S0 and concentrated under reduced pressure. The resulting crude was purified by silica gel column (100- 200 mesh, elution: 30% EtOAc in Hexane) chromatography to afford the title compound as off white solid. Wt: 0.500 g: Yield: 72.6%; Ή NMR (300 MHz, CD3OD): δ 3.75-3.704 (m, IH), 2.55-2.52 (m, IH), 2.19-2.06 (m, 2H), 1.43 (s, 9H), 1.25 (s, 3H), 0.92 (s, 3H); Mass: [M+Na]+ 266 (100%); IR (KBr, cm'1): 3300, 3249,
2967, 1701, 1643, 1479, 1403, 1265, 1159, 1109, 1029, 858, 778; M.R: 142.8 °C - 149.0 °C.
Intermediate 4: Synthesis of (lR,3S)-3-(benzyloxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid:
azide:
Figure imgf000029_0002
A stirred solution of (lS,3R)-3-acetyl-2,2-dimethylcyclobutanecarboxylic acid (step 1 for Intermediate 1, about 1.0 g, 5.88 mmol) in acetone (20 ml) at about
0°C triethyl amine (about 1.6 ml, 11.76 mmol) and ethyl chloroformate (about 1.12 ml, 1 1.76 mmol) were added slowly by portion wise and the reaction was stirred at about 0°C for about 3 hours. After completion of the reaction (monitored by TLC), NaN3 (about 0.84 g, 12.93 mmol) was dissolved in minimum amount of water and added to reaction mixture at about 0°C then the reaction mixture was allowed to stir at room temperature for about 1 .5 hours. After completion of the reaction, the reaction mixture was extracted with DCM and washed with water, brine and dried with Na2S04 and benzyl alcohol (about 0.95 ml, 8.82 mmol). The resulting crude product was proceeding for next reaction immediately without characterization (considering as 100% yield proceeded for next reaction). Crude wt: 1. 147 g ( 100% yield).
Step 2: Synthesis of benzyl ( lS,3R ethylcyclobut} lcarbaniate:
Figure imgf000030_0001
A stirred solution of ( l S,3R)-3-acetyl-2,2-dimethylcyclobutanecarbonyl azide (Step 1 , about 1. 147 g, 5.88 mmol) in toluene (20 ml) at 0 °C, benzyl alcohol (about
0.95 ml, 8.82 mmol) was added and stined at reflux temperature for about 4 hours. After completion of the reaction (monitored by TLC), the solvent was evaporated under reduced pressure. The resulting crude was purified by silica gel column (60/120mesh, elution: 5% EtOAc in Hexane) chromatography to afford the title compound as a liquid. Wt: 1.3 g; Yield: 81.2 %;Ή NMR (300 MHz, CDC13): δ 7.38-
7.29 (m, 5H), 5.08 (q, 2H, 7 = 12 Hz), 4.80 (d, 1 H, J = 8.0 Hz), 3.92 (q, 1 H, J = 9.3 Hz), 2.77-2.7 1 (m, 1 H), 2. 16-2.08 (m, 1 H), 2.07 (s, 3H), 1 .40 (s, 3H), 0.82 (s, 3H); Mass: [M+ l ]+ 276 ( 10%), [M+Na]+ 298 ( 100%); IR (KBr, cm" 1 ): 3338, 2956, 2927, 1703, 1648, 1530, 1460, 1370, 1282, 1254, 1 183, 1042, 944.
Step 3: Synthesis of (lR,3S)-3-(benzy!oxycarbonyla ino)-2,2- dimethylcyclobutanecarboxylic acid:
A stirred solution of benzyl ( l S,3R)-3-acetyl-2,2- dimethylcyclobutylcarbamate (step 8, about 1 .3 g, 4.72 mmol) in dioxane (90 ml): water (30 ml) aqueous NaOBr solution (about 80 ml) was added slowly at about -5
°C and stirred at the same temperature for about 5 hours. After completion of the reaction (monitored by TLC), the reaction mixture was washed with methyl t-butyl ether and the aqueous layer was adjusted pH (2-3) with I N HC1 and extracted with EtOAc, dried with Na?S04 and the solvent was evaporated. The resulting crude was purified by silica gel column ( 100-200 mesh, elution: 20% EtOAc in Hexane) chromatography to afford the title compound as a semi solid. Wt: 1 .2 g: Yield: 92%. Ή NMR (300 MHZ, CDC13): δ 7.38-7.31 (m, 5H), 5.09 (q, 2H, J = 10.8 Hz), 4.89 (d, 1 H, J = 8.4 Hz), 3.98-3.89 (m, 1 H), 2.64-2.53 (m, 1 H), 2.5 1 -2.26 (m, 2H), 1.32 (s, 3H), 0.98 (s, 3H); IR (KBr, cm" 1 ): 341 1 , 3351 , 2961 , 1705, 1532, 1456, 1412, 1343, 1256, 1044, 1003, 776, 697; Mass: [M+Na]+ 300 ( 100%).
NaOBr Solution: A solution of NaOH (about 9.4 g, 233.6 mmol) in H20 ( 180 ml) Br2 (3.3 ml, 63.72 mmol) was added at about 0°C slowly then the yellow colored NaOBr solution formed which was used immediately for the reaction.
Intermediate 5: Synthesis of ( l R,3S)-3-(methoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid:
Figure imgf000031_0001
Step 1 : Synthesis of ( I R,3S)-methyl 3-(benzyloxycarbonylamino)-2,2- dimethylcyclob tanecarboxylate:
Figure imgf000031_0002
A stirred solution of ( l R,3S)-3-(benzyloxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 4, about 1.3 g, 4.69 mmol) in DMF ( 10 ml) Cs2CO? ( 1 .987 g, 6. 10 mmol) and Mel (0.32 ml, 5. 16 mmol) were added slowly by portion wise at about 0°C, after addition the reaction mixture was stirred at room temperature for about 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc ( 100 ml) and washed with water (50 ml), brine (50 ml), dried with Na2S04. The solvent was evaporated and purified by silica gel column (60- 120, elution 10% EtOAc in hexane) chromatography to afford the title compound as pale yellow syrup. Wt: 1 .0 g: Yield: 76.9%: Ή NMR (300 MHz, CDjOD): δ 7.39-7.24 (m, 5H), 5.05 (s, 2H),
3.87-3.73 (m, 1 H), 3.65 (s, 3H), 2.66-2.58 (m, 1 H), 2.24-2. 13 (m, 2H), 1 .24 (s, 3H), 0.86 (s, 3H); Mass: [M+Na]+ 3 14 ( 100%).
Step 2: Synthesis of ( 1 R,3S)-methyI 3-(metho.xycarbonylamino)-2,2- dimethylcyclobntanecarboxylate:
Figure imgf000032_0001
A stirred solution of ( l R,3S)-methyl 3-(benzyloxycarbonylamino)-2,2- dimethylcyclobutanecarboxylate (step 1 , about 1.0 g, 3.436 mmol) in EtOH ( 15 ml) at room temperature 10 % Pd/C (catalytic amount) was added and the reaction mixture was stirred under H2 gas atmosphere at same temperature for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through celite bed and the obtained filtrate concentrated under reduced pressure to afford ( l R,3S)-methyl 3-amino-2,2-dimethylcyclobutanecarboxylate as a liquid, which was dissolved in DCM at about 0°C then triethylamine (about 1 .20 ml, 8.591 mmol) was added followed by methyl chloroformate (about 0.3 17 ml, 4. 123 mmol) added and allowed to stir at room temperature for about 12 hours. After completion of the reaction (monitored by TLC), the solvent was evaporated and purified by silica gel column ( 100-200 mesh, elution 10% EtOAc in hexane) chromatography to afford the title compound as a pale yellow syrup. Wt: 0.6 g; Yield: 87.7%; Ή NMR (300 MHz, CD3OD): δ 4.76 (brd, 1 H), 3.96-3.84 (m, 1 H),
3.73 (s, 3H), 3.69 (s, 3H), 2.62-2.54 (m, 1 H), 2.41 -2.32 (m, 1 H), 2. 12-2.01 (m, 1 H), 1 .29 (s, 3H), 0.91 (s, 3H);Mass: [M+ l ]+ 216 ( 100%), [M+Na]+ 238 (25%).
Step 3: Synthesis of ( lR,3S)-3-(methoxycarbonyla ino)-2,2- dimethylcyclobutanecarboxylic acid:
To a stirred solution of ( l R,3S)-methyl 3-(methoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylate (Step 2, about 0.6 g, 2.79 mmol) in tetrahydrofuran (20 ml) NaOH (about 0.25 M) was added at about 0°C and allowed to stir at room temperature for about 1 hour. After completion of the reaction (monitored by TLC), the tetrahydrofuran was evaporated, acidified with I N HC1 and extracted with
EtOAc. The organic layer was dried with Na2S04 and the solvent was evaporated. The resulting crude was purified by silica gel column ( 100-200 mesh, elution: 30% EtOAc in Hexane) chromatography to afford the title compound as a semi solid. Wt: 0.37 g; Yield: 66.0 %; Ή NMR (300 MHZ, CD3OD): δ 8.69 (d, 1 H, J = 7.5 Hz), 3.84-3.75 (m, 1 H), 3.61 (s, 3H), 2.61 -2.50 (m, 1 H), 2.25-2.08 (m, 2H), 1 .25 (s, 3H),
0.92 (s, 3H); IR (KBr, cm 1 ): 3373, 3324, 2962, 1702, 1557, 1538, 1463, 1429, 1339, 1290, 1276, 1 197, 1057, 984, 93 1 , 780; Mass: [M+ l ]+ 202 (20%), (M+Na]+ 224 ( 100%). Intermediate 6: Synthesis of ( l R,3S)-3-(tert-butoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid:
Figure imgf000033_0001
Step 1 : Synthesis of (lR,3S)-methyl 3-(tert-butoxycarbony\amino)-2,2- dimethylcyclobutanecarboxylate:
Figure imgf000033_0002
To a solution of ( l R,3S)-methyl 3-(benzyloxycarbonylamino)-2,2- dimethylcyclobutanecarboxylate (Step 1 for Intermediate 5, about 4.7 g, 16. 151 mmol) in EtOAc (20 ml), palladium on carbon (catalytic amount) was added and stirred under H2 gas atmosphere for about 3 hours at room temperature. After completion of the reaction, the reaction mixture was filtered through celite bed, the filtrate was concentrated and dried under vacuum then dissolved in DCM and cooled to about 0 °C then triethylamine (about 5.3 ml, 38.216 mmol) followed by (Boc):0 (about 3.39 ml, 19. 108) were added and allowed to stir for about 12 hours at room temperature. After completion of the reaction (monitored by TLC) the reaction mass was diluted with DCM, washed with water and brine. The organic layer was dried over Na2S04 and the solvent was evaporated, the resulting crude purified by silica gel column ( 100-200 mesh, elution: 3% EtOAc in Hexane) to afford the title compound as an off white solid. Wt: 1 .5 g; Yield: 36. 14 %; Ή NMR (300 MHz,
CD3OD): δ 3.78-3.69 (m, 1 H), 3.65 (s, 3H), 2.66-2.57 (m, 1 H), 2.24-2. 10 (m, 2H), 1 .43 (s, 9H), 1 .24 (s, 3H), 0.86 (s, 3H); IR (KBr, cm" 1 ): 3404, 3384, 2957, 1706, 1522, 1462, 1439, 1367, 1343, 1266, 1 165, 1008, 984, 871 , 778; Mass: [M+Na]+ 280 ( 100%); M R: 101 .2- 103.5 °C.
Step 2: Synthesis of ( 1 R,3S)-3-(tert-butoxycarbonylainino)-2,2- dimethyicyclobutanecarboxylic acid:
To a stirred solution of ( l R,3S)-methyl 3-(tert-butoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylate (step 1 , about 1 .5 g, 5.836 mmol) in tetrahydrofuran (60 ml) at about 0 °C sodium hydroxide (0.25 M, 46.6 ml) was added and stirred at room temperature for about 2 hours. After completion of the reaction (monitored by TLC), the solvent was evaporated, acidified with citric acid and extracted with EtOAc. The organic layer was dried over Na2S04 and the solvent was evaporated, the resulting crude residue was recrystalized in DCM and hexane to afford the title compound as an off white solid. Wt: 1.1 g; Yield: 78.5 %; Ή NMR (300 MHz, CD3OD): δ 3.79-3.62 (m, 1H), 2.62-2.50 (m, 1H), 2.21-2.02 (m, 2H), 1.43 (s, 9H), 1.25 (s, 3H), 0.91 (s, 3H); IR (KBr, cm"1): 3303, 3251, 2967, 1702, 1645, 1457, 1402, 1367, 1264, 1159, 1109, 1049, 857, 731; Mass: [M-l]" 242
(100%), [M+Na]+266(100%);M.R: 129.4-132.5 °C.
Intermediate 7: Synthesis of (2S,3S)-3-amino-4-phenyl-l-(l-(4-(pyridin-2- yl)benzyl)hydrazinyl)butan-2-ol:
Figure imgf000034_0001
Step 1: Synthesis of tert -butyl 2-((2S,3S)-3-(tert-butoxycarbonylamino)-2-hydroxy-4- phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hydrazinecarboxylate:
Figure imgf000034_0002
A stirred solution of tert-butyl 2-(4-(pyridin-2- yl)benzyl)hydrazinecarboxylate (about 1.3 g, 4.347 mmol) and tert-butyl (S)-1-((R)- oxiran-2-yl)-2-phenylethylcarbamate (about 1.7 g, 6.521 mmol) in Isopropanol was refluxed for about 24 hours. After completion of the reaction (monitored by TLC), the reaction mixture brought to room temperature and water (23 ml) was added and stirred at same temperature for about 16 hours. Resulting solid precipitation was filtered and washed with water (23 ml), cold methyltertiarybutyl ether (3 ml) then dissolved in acetonitrile, water (35 ml) was added and filtered, the resulting solid purified by silica gel column (100-200 mesh, elution: 20% EtOAc in Hexane) to afford the title compound as an off white solid. Wt:1.5 g; Yield: 62.5 %; Ή NMR (300 MHz, CDCI3): δ 8.69 (d, 1H, J = 4.5 Hz,), 7.95 (d, 2H, J = 8.1 Hz), 7.76-7.69 (m, 2H), 7.41 (d, 2H, J = 8.1 Hz), 7.25-7.23 (m, 6H), 5.25 (s, 1H), 5.12 (d, 1H, J =
9.3 Hz), 4.54 (s.1H).4.07-3.94 (m, 2H), 3.68-3.55 (m, 2H).2.95-2.80 (m, 3H), 2.46 (d, 1H, J= 10.2 Hz), 1.38 (s, 9H), 1.32 (s, 9H); Mass: [M+H}+ 563 (3%), [M+Na) + 585 (100%). Step 2: Synthesis of (2S,3S)-3-amino-4-phenyl-l-(]-(4-(pyridin-2- yl)benzyl)hydrazinyl)butan-2-ol. hydrogen chloride salt:
To a stirred solution of tert-butyl 2-((2S,3S)-3-(tert-butoxycarbonylamino)-2- hydroxy-4-phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hydrazinecarboxylate (step 1 , about 0.5 g, 0.889 mmol) in tetrahydrofuran (2.5 ml) concentrated HCl (0.339 ml) was added drop wise at room temperature to give colored solution. The reaction mixture was stirred at about 45-55 0°C for about 3 hours. After completion of the reaction tetrahydrofuran layer was removed and the remaining oil compound rinsed with tetrahydrofuran ( 1 ml) then it was used as such for further reactions.
Intermediate 8: Synthesis of methyl (S)- l -(2-((2S.3S)-3-amino-2-hydroxy-4- phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hydrazinyl)-3.3-dimethyl- l -oxobutan-2- ylcarbamate:
Figure imgf000035_0001
Step 1: Synthesis of (S)-2-(methoxycarbonylaniino)-3,3-dimethylb tanoic acid:
Figure imgf000035_0002
A stirred solution of (S)-2-amino-3,3-dimethylbutanoic acid (about 5.0 g, 38. 16 mmol) in Dioxane (about 20 ml) and sodium hydroxide (2N, 62 ml, PH = 8-9) at about 0 °C, methychlroformate (about 5.88 ml, 76.33 mmol) was added drop wise and stirred at about 60 °C for about 18 hours. The reaction mixture was cooled to room temperature, extracted with DCM, the aqueous layer was separated and acidified with I N HCl. The resulting solution was extracted with EtOAc, dried over Na2S04 and the solvent was evaporated under reduced pressure. The resulting crude was stirred in hexane and decants to afford the title compound as a solid. Wt: 8.5 g: Yield: quantitative; Ή NMR (300 MHZ, CDCI3): δ 5.25 (d, 1 H, J = 10.5 Hz), 4. 19
(d, 1 H, J = 9.6 Hz) 3.70 (s, 3H), 1 .03 (s, 9H); Mass: [M- l ]' 188 ( 100%); IR ( Br, cm 1 ): 3379, 2974, 1727, 1688, 1546, 1466, 1332, 1263. 121 1 , 1070, 1034, 1018, 843, 696. Step 2: Synthesis of (S)-tert-butyl 2-(2-(methoxycarbonyla ino)-3,3- dimethylb tanoyljhydrazinecarboxylate: NH-NHBoc
o
To a stirred solution of (S)-2-(methoxycarbonylamino)-3,3-dimethylbutanoic acid (step 1 , about 8.50 g, 44.97 mmol) in EtOAc ( 1 10 ml) at about 0 °C l -Ethyl-3- (3-Dimethylaminopropyl)carbodiimide (9.48 g, 49.47 mmol), hydroxybenzotnazole (7.57 g, 49.47 mmol) and N-methyl morpholine (5.95 ml, 53.95 mmol) were added, after 30 minutes t-Butyl carbazate (about 6.53 g, 1 .423 mmol) was added and stirred at room temperature for about 18 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc and washed with water, saturated NaHCCh solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column ( 100-200 mesh, elution 35% EtOAC in hexane) to afford the title compound as an off white solid. Wt: 9.8 g; Yield: 72%; Ή NMR (300 MHz, CDC13): δ 8.83 (brs, 1 H), 6.85 (brs, 1 H), 5.71 (d, 1 H, J = 9.9 Hz), 4. 15 (d, 1 H, J = 9.6 Hz), 3.66 (s, 3H), 1 .44 (s, 9H), 1 .03 (s, 9H); Mass: [M+Na]+ 326 ( 100%); IR (KBr, cm" 1 ): 3303, 2978, 1709, 1681 , 1534, 1394, 1239. 1 162, 1060, 1020, 863, 812, 778; M.R: 65.4 °C -67.6 °C.
Step 3: Synthesis of (S)-niethyl 1 -hydrazinyl-3, 3 -dimethyl- 1 -oxob tan-2- xlcarbamate:
Figure imgf000036_0001
A solution of (S)-tert-butyl 2-(2-(methoxycarbonylamino)-3,3- dimethylbutanoyl)hydrazinecarboxylate (step 2, about 2.6 g, 8.58 mmol) in 2N HC1 in dioxane (20 ml) stirred at about 0 °C to room temperature for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated and basified with solid NaHC03 then extracted with EtOAc and dried over
Figure imgf000036_0002
the solvent was evaporated and resulting crude proceeded for next reaction without characterization. Wt: 1 .56 g: Yield: 90 %.
Step 4: synthesis of (S,E)-methyl 3,3-di ethyl-l -oxo-l -(2-(4-(pyridin-2- yl)benzylidene)hydrazinyl)buta -2-ylcarbamate:
Figure imgf000036_0003
A solution of (S)-methyl l -hydrazinyl-3,3-dimethyl- l -oxobutan-2- ylcarbamate (step 3, about 1.56 g, 7.722 mmol) and 4-(pyridin-2-yl)benzaldehyde (about 1 .407 g, 7.722 mmol) in isopropyl alcohol stirred at reflux temperature for about 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated and purified by silica gel column ( 100-200 mesh, elution
35% EtOAc in hexane) to afford the compound as a semi solid. Wt: 2.7 g: Yield: 95.7%; Ή NMR (300 MHz, CD3OD): δ 8.65 (d, 1 H, J = 5. 1 Hz), 8.21 (s, 1 H), 8.07- 7.90 (m, 6H), 7.49-7.42 (m, 1 H), 4.04 (s, 1 H), 3.66 (s, 3H), 1 .06 (s, 9H); Mass: [M+ l ]+ 369 (35%), [M+Na]+391 ( 100%); IR (KBr, cm" 1): 3434, 3325, 2967, 1671 , 1532, 1467, 1370, 1236, 1 175, 1078, 1062, 1015, 781 .
Step 5: Synthesis of (S)-methyl 3,3-dimethyl-l -oxo- l-(2-(4-(pyridin-2- yl)benzyl)hydrazinyl)butan-2-ylcarbamate:
Figure imgf000037_0001
To a solution of (S,E)-methyl 3,3-dimethyl- l -oxo- l -(2-(4-(pyridin-2- yl)benzylidene)hydrazinyl)butan-2-ylcarbamate (step 4, about 3.0 g, 8. 16 mmol) in EtOH (20 ml) sodiumformate (about 0.99 g, 14.673 mmol) was dissolved in minimum amount of water and palladium on carbon ( 10%) were added and stirred under H2 gas atmosphere at room temperature for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through celite bed and the obtained filtrate was concentrated under reduced pressure. The resulting crude purified by silica gel column ( 100-200 mesh, elution 50% EtOAc in hexane) to afford the title compound as a semi solid. Wt: 2. 1 g: Yield: 70 %; Ή NMR (300 MHz, CDClj): δ 8.68 (d, 1 H, J = 4.5 Hz), 7.97-7.92 (m, 3H), 7.78-7.68 (m. 3H), 7.44 (d, 2H, 7 = 8. 1 Hz), 7.27-7. 19 (m, 1 H), 5.56 (d, 1 H, J= 9.6 Hz), 4.02 (dd, 2H, J
= 1 1 .4, 17.7 Hz), 3.88 (d, 1 H, J = 9.6 Hz), 3.63 (s, 3H), 0.96 (s, 9H); Mass: [M+ l ]+ 37 1 (40%), [M+Na]+393 ( 100%); IR (KBr, cm 1 ): 3295, 2959, 1710, 1659, 1588, 1532, 1467, 1435, 1368, 13 19, 1240, 1055, 1017, 777. Step 6: Synthesis of methyl (S)-l -(2-((2S,3S)-3-(tert-butoxycarbonylamino)-2- hydrox)'-4^henylbuty1)-2-(4-(pyridin-2-yl)benzyl)hydrazinyl)-3 -dimethy oxobutan-2-ylcarbamate:
Figure imgf000038_0001
A stirred solution of (S)-methyl 3,3-dimethyl-l-oxo-l-(2-(4-(pyridin-2- yl)benzyl)hydrazinyl)butan-2-ylcarbamate (step 5, about 2.1 g, 5.675 mmol) and tert-butyl (S)-l-((R)-oxiran-2-yl)-2-phenylethylcarbamate (about 2.23 g, 8.513 mmol) in isopropanol (20 ml) was refluxed for about 16 hours. After completion of the reaction (monitored by TLC), the solvent was evaporated and the resulting crude was purified by silica gel column (100-200 mesh, elution: 40% EtOAc in hexane) to afford the title Compound as an off white solid. Wt: 1.1 g; Yield: 30.8%; Ή NMR (300 MHz, CDCI3): δ 8.69 (d, 1H, J = 4.2 Hz), 7.94 (d, 2H, J = 8.1 Hz), 7.80-7.68 (m, 2H), 7.43 (d, 2H, J = 8.1 Hz), 7.33-7.12 (m, 6H), 6.40 (s, 1H), 5.24 (d, 1H, J =
9.0 Hz), 5.07 (d, 1H, J = 9.6 Hz), 4.73 (s, 1H), 4.12 (d, 1H, J = 14.1 Hz), 3.92 (d, 1H, J = 13.8 Hz).3.71-3.48 (m, 6H), 2.99-2.84 (m, 3H), 2.60 (d, 1H, 12.3 Hz), 1.35 (s, 9H), 0.72 (s, 9H); Mass: [M+H]+633 (25%), [M+Na]+656 (100%); R ( Br, cm '): 3400, 2965, 1693, 1656, 1513, 1467, 1367, 1327, 1248, 1170, 1054, 1019, 779, 749, 700, 640; M.R: 177.3 °C -180.2 °C.
Step 7: Synthesis of methyl (S)-l -(2-((2S,3S)-3-amino-2-hydroxy-4-phenylb tyl)-2- (4-(pyridin-2-yl)benzyl)hydraz.inyl)-3, 3 -dimethyl- 1 -oxobittan-2-y /carbamate:
To a stirred solution of methyl (S)-l -(2-((2S,3S)-3-(tert- biuoxycarbonylamino)-2-hydroxy>-4-phenylb ti)-2-(4-(pyridin-2- yl)benzyl)hydrazinyl)-3,3-dimethyl-l -oxobutan-2-ylcarbamate (Step 6, about 0.4 g, 0.632 mmol) in tetrahydrofuran (5.0 ml) at about 0°C 4N HCl (4 ml) was added drop wise and stirred at about 50 °C for about 3 hours. After completion of the reaction the solvent was removed and dissolved in EtOH then concentrated and dried under vacuum then it was used as such for further reactions. ntermediate 9: Synthesis of methyl (S)-l-(2-((2S,3S)-3-amino-2-hydroxy-4- phenylbutyl)-2-benzylhydrazinyr)-3,3-dimethyl-l-oxobutan-2-ylcarbamate:
Figure imgf000038_0002
Step 1: Synthesis of (S,E)-methyl l-(2-benzylidenehydrazinyl)-3,3-dimethyl-l- oxob tan-2-ylcarbamate: 0
A solution of (S)-methyl l -hydrazinyl-3,3-dimethyl- l -oxobutan-2- ylcarbamate (Step 3 for Intermediate 8, about 1 .8 g, 8.866 mmol) and benzaldehyde (about 0.93 g) in isopropyl alcohol at room temperature was stirred for about 1.5 hours. After completion of the reaction (monitored by TLC) the reaction mixture was concentrated and purified by silica gel column ( 100-200 mesh, elution 35% EtOAc in hexane) to afford the title compound as a pale yellow syrup. Wt: 2.4 g: Yield: 93%; Ή NMR (300 MHz, CDC13): δ 9.04 (s, 0.6H), 8. 12 (s, 0.3H), 7.79 (m, 2H), 7.45-7.32 (m, 3H), 5.65 (d, 0.3H, J = 9.9 Hz), 5.52 (d, 0.6H), 5.33 (d, 0.7H, J = 9.6 Hz), 4.02 ((d, 0.3H, J = 9.6 Hz), 3.70, 3.68 (2s, 3H), 1 .08, 1 .06 (2s, 9H); Mass:
[M+ 1 ]+ 292 (50%), [M+Na]+3 14 ( 100%).
Step 2: Synthesis of (S)-methyl 1 -(2-benzylhydrazinyl)-3,3-dimethyl-J -oxobutan-2- vlcarbamate
Figure imgf000039_0001
A solution of (S,E)-methyl l -(2-benzylidenehydrazinyl)-3,3-dimethyl- l - oxobutan-2-ylcarbamate (step 1 , about 2.4 g, 8.24 mmol) in EtOH (40 ml), HCCbNa ( 1 .0 g, 14.84 mmol) was dissolved in minimum amount of water and palladium on carbon ( 10%) were added and stirred under H? gas atmosphere at room temperature for about 12 hours. After completion of the reaction (monitored by TLC) the reaction mixture was filtered through celite bed and the obtained filtrate was concentrated under reduced pressure. The resulting crude was purified by silica gel column ( 100-200 mesh, elution 35% EtOAc in hexane) to afford the title compound as a semi solid. Wt: 2. 1 g: Yield: 87 %; Ή NMR (300 MHZ, CDCI3): δ 7.49 (s, 1 H), 7.38-7.22 (m, 5H), 5.45 (d, 1 H), 4.92 (brs, 1 H), 3.99-3.91 (m, 2H), 3.80 (d, 1 H, J =
9.6 Hz), 3.65 (s, 3H), 0.96 (s, 9H); Mass: [M+ l ]+ 294 (50%), [M+Na]+316 ( 100%); IR (KBr, cm" 1 ): 3298, 2962, 1712, 1656, 1456, 1369, 1243. 1055, 1020, 846, 777, 744, 698. Step 3: Synthesis of methyl (S)-l -(2-benzyl-2-((2S S)-3-(tert- bιιtoxycarbonylami o)-2 ιydroxy-4ψhenylbι^tΛ1)hydrazinyl)-3 -dimethyl- l - oxobu tan -2 -y lea rba mate:
Figure imgf000040_0001
A stirred solution of (S)-methyl l -(2-benzylhydrazinyl)-3,3-dimethyl- l - oxobutan-2-ylcarbamate (step 2, about 2. 1 g, 7.167 n mol) and tert-butyl (S)- 1 -((R)- oxiran-2-yl)-2-phenylethylcarbamate (about 2.82 g, 10.750 mmol) in isopropanol (20 ml) was refluxed for about 16 hours. After completion of the reaction (monitored by TLC) the solvent was evaporated and the resulting crude was purified by silica gel column ( 100-200 mesh, elution: 30% EtOAc in Hexane) to afford the title compound as an off white solid. Wt:2.5 g; Yield: 62.8 %; Ή NMR (300 MHz, CDCI3): δ 7.36-7. 12 (m, 10H), 6.26 (s, 1 H), 5.21 (d, 1 H), 5.05 (d, 1 H, J = 9.3 Hz), 4.69 (s, 1 H), 4.07 (d, 1 H, J = 13.8 Hz), 3.85 (d, 1 H, J = 13.8 Hz), 3.66 (s, 3H), 3.58- 3.42 (m, 3H), 2.95-2.83 (m, 3H), 2.59-2.5 1 (m, 1 H), 1.37 (s, 9H), 0.72 (s, 9H); Mass: [M+H]+ 556 (25%), [M+Na]+ 579 ( 100%); BR (KBr, cm 1 ): 3401 , 3294, 2978, 2964, 1695, 1655, 1509, 1457, 1446, 1367, 1326, 1246, 1 171 , 1 154, 1018, 794, 747,
698; M.R: 1 81.8 UC - 184.9 X.
Step 4: Synthesis of methyl (S)-l -(2-((2S,3S)-3-amino-2-hydroxy-4-phenylbutyl)-2- benzylhydrazinyl)-3, 3 -dimethyl- 1 -oxobutan-2-ylcarbamate :
A stirred solution of methyl (S)-l -(2-benzyl-2-((2S,3S)-3-(tert- butoxycarbonylam\no)-2-hydroxy-4-phenylbutyl )hydrazinyl)-3,3-dimethyl- 1 - oxobutan-2-ylcarbamate (Step 3, about 0.5 g, 0.889 mmol) in tetrahydrofuran (6.0 mJ) at about 0°C 4N HCl (4 ml) was added drop wise and stirred at about 50 °C for about 3 hours. After completion of the reaction the solvent was removed and dried under vacuum then it was used as such for further reactions. Intermediate 10: Synthesis of methyl (S - l -(2-((2S,3S)-3-amino-2-hydroxy-4- phenylbuty )-2-(biphenyl-4-ylmethyr)hydrazinyl)-3,3-dimethyl- l -oxobutan-2- ylcai bamate:
Figure imgf000040_0002
Step 1: Synthesis of biphenyl-4-carbaldehyde :
Figure imgf000041_0001
A stirred solution of phenylboronic acid (about 1.5 g, 12.302 mmol) and 4- bromo benzaldehyde (about 2.048 g, 1 1.071 mmol) in ethanol: toluene (4:3) (about 18 ml) was degassed with N2 then Pd(Ph3P)4 in ethanol: toluene (4:3) (about 3 ml) solution and 3M Na2C03 (8.7 ml) were added and the reaction mass was stirred at about 70 °C for about 3 hours. After completion of the reaction, the reaction mass was diluted with EtOAc and washed with water, brine and dried over Na2S0.4 and the resulting crude was purified by silica gel column ( 100-200 mesh, elution 2% EtOAc in hexane) to afford the title compound as a liquid. Wt: 1.9 g: Yield: 86.3%; Ή NMR (300 MHz, CDC13): δ 10.07 (s, 1 H), 7.96 (d, 2H, J = 8. 1 Hz), 7.76 (d, 2H, J = 8.4 Hz), 7.64 (d, 2H, J = 7.8 Hz), 7.53-7.39 (m, 3H); IR (KBr, cm" 1 ): 3031 , 283 1 . 1704, 1604, 1581 , 1450, 1214, 1 169, 1 109, 837, 762, 696.
Step 2: Synthesis of (S,E)-methyl l -(2-(biphenyI-4-ylmethylene)hydrazinyl)-3,3- dimethyl-l -oxobutan-2-ylcarbamate:
Figure imgf000041_0002
A solution of (S)-methyl l -hydrazinyl-3,3-dimethyl- l -oxobutan-2- ylcarbamate (Step 3 for Intermediate 8, about 1 .67 g, 8.222 mmol) and biphenyl-4- carbaldehyde (step 1 , about 1 .49 g, 8.226 mmol) in isopropanol (20 ml) was stirred at reflux temperature for about 16 hours. After completion of the reaction (monitored by TLC) water was added then the solid was precipitated. The resulting crude was filtered and dried under vacuum. Wt: 2.7 g: Yield: 90%; Ή NMR (300 MHz, CD3OD): δ 7.87 (d, 2H, J = 8. 1 Hz), 7.74-7.62 (m, 5H), 7.52-7.31 (m, 3H), 4.05 (d, 1 H, J = 8. 1 Hz), 3.67 (s, 3H), 1.06 (s, 9H); Mass: [M+ l ]+ 368 ( 15%), [M+Na]+390 ( 100% ).
Step 3: synthesis of (S)-methyl l -(2-(biphenyl-4-ylmethyl)hydrazinyl)-3,3-dimethyl- l -oxobutan-2-xlcarbaniate:
Figure imgf000041_0003
To a stirred solution of (S,E)-methyl l -(2-(biphenyl-4- ylmethylene)hydrazinyl)-3,3-dimethyl- l -oxobutan-2-ylcarbamate (step 2, about 2.7 g, 7.356 mmol) in ethanol (30 ml) HC02Na (0.90 g, 13.24 mmol) dissolved in minimum amount of water and Pd/C ( 10%) were added and stirred under H? gas atmosphere at room temperature for about 2 hours. After completion of the reaction
(monitored by TLC), the reaction mixture was filtered through celite bed and the obtained filtrate was concentrated under reduced pressure. The resulting crude was purified by silica gel column ( 100-200 mesh, elution 40% EtOAc in hexane) to afford the title compound as a semi solid. Wt: 2.0 g: Yield: 74 %; Ή NMR (300 MHz, CDC13): δ 7.94 (s, 1 H), 7.60 (m, 4H), 7.49-7.29 (m, 5H), 5.56 (d, 1 H, J = 9.6
Hz), 4.03-3.91 (m, 2H), 3.89 (d, 1 H, J = 9.6 Hz), 3.62 (s, 3H), 0.97 (s, 9H); Mass:
[M+Na]+392 ( 100%); IR (KBr, cm" 1): 3259, 2988, 2968, 1727, 1646, 1522, 1467, 1374, 1304, 1247, 1056, 957, 823; M.R: 136.2 °C - 142.5 °C.
Step 4:Synthesis of methyl <S)-l-(2-(biphenyl-4-yhnethyl)-2-((2S, 3S)-3-(tert- butoxycarbony\amm' o)-2-hydroxy-4-phenylbuty\)hydraz\ny\)-3
oxobu tan-2-ylca rbamate :
Figure imgf000042_0001
A stirred solution of (S)-methyl l -(2-(biphenyl-4-ylmethyl)hydrazinyl)-3,3- dimethyl- l -oxobutan-2-ylcarbamate (step 3, about 1 .5 g, 4.065 mmol) and tert-butyl
(S)- l -((R)-oxiran-2-yl)-2-phenylethylcarbamate (about 1 .06 g, 4.065 mmol) in isopropanol (30 ml) was refluxed for about 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was brought to room temperature and ice was added then the solid formed was filtered, washed with water and dried under vacuum. Wt: 2.3 g; Yield: 90%; Ή NMR (300 MHz, CDCI3): δ 7.59-7.50 (m,
4H), 7.49-7.3 1 (m, 5H), 7.29-7. 13 (m, 5H), 6.44 (s, 1 H), 5.27 (d, 1 H, J = 9.3 Hz), 5. 10 (d, 1 H), 4.76 (s, 1 H), 4. 12 (d, 1 H, J = 13.5 Hz), 3.89 (d, 1 H, 13.5 Hz), 3.64 (s, 3H), 3.7 1 -3.47 (m, 3H), 2.98-2.83 (m, 3H), 2.64-2.57 (m, 1 H), 1.36 (s, 9H), 0.7 1 (s, 9H); Mass: [M+Na]+ 655 ( 100%); IR (KBr, cm" 1 ): 3397, 3030, 2965, 1692, 1655, 15 13, 1456, 1392, 1367, 1428, 1 17 1 , 1053, 1020, 762, 747; M.R: 199.5 °C -201 °C. Step 5: Synthesis of methyl (S)-l -(2-((2S,3S)-3-amino-2-hydroxy-4-phenylbutyl)-2- ( biphenyl-4-ylmethyl)hydrazinyl)-3, 3 -dimethyl- 1 -oxobutan-2-ylcarbamate:
A stirred solution of methyl (S)-l-(2-(biphenyl-4-ylmethyl)-2-((2S,3S)-3- (tert-bittox carbonylamino)-2-hydrox '-4-phenylbut)'l)hydrazinyl)-3,3-di
oxob tan-2-ylcarbamate (Step 4, about 0.3 g, 0.474 mmol) in tetrahydrofuran (6.0 ml) at about °C 4N HCl (4 ml) was added drop wise and stirred at about 50 °C for about 2 hours. After completion of the reaction the solvent was removed and dissolved in EtOH then concentrated and dried under vacuum then it was used as such for further reactions.
Intermediate 1 1 : Synthesis of methyl (S)- l -(2-((2S,3S )-3-amino-2-hydroxy-4- phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hydrazinyl)-4-methyl- l -oxopentan-2- ylcarbamate:
Figure imgf000043_0001
Step 1: Synthesis of ( S )-2 -( methoxycarbonylamino)-4-methylpentonoic acid:
Figure imgf000043_0002
A stirred solution of (S)-2-amino-4-methylpentanoic acid (about 5.0 g, 38. 16 mmol) in Dioxane (20 ml) at 0 °C 2N NaOH (62 ml, PH = 8-9) and methychloioformate (5.88 ml, 76.33 mmol) were added drop wise and stirred at about 60 °C for about 18 hours. The reaction mixture was cooled to room temperature, extracted with DCM, the aqueous layer was separated and acidified with I N HCl. The resulting solution was extracted with EtOAc, dried over Na2S04 and the solvent was evaporated under the reduced pressure and the resulting crude was stirred in he.xane and decanted to afford the title compound as a semi solid. Wt: 8.0 g: Yield: quantitative; Ή NMR (300 MHz, CDC13): δ 5.09 (d, 1 H, J = 8.4 Hz), 4.43-4.3 1 (m, 1 H), 3.7 1 (s, 3H), 1 .80- 1 .39 (m, 3H), 0.96 (d, 6H, J = 6. 1 Hz); Mass: [M+H]+ 190 (60%) [M+Na]+ 2.12 ( 100%); IR (KBr, cm" 1 ): 3354, 2960, 1709, 1540, 1467, 1456, 1370, 1270, 1235, 1 123, 1058, 924, 781 .
Step 2: Synthesis of (S)-tert-b t)'l 2-(2-(methoxycarbonylamino)-4-methylpentanoyl) hydrazinecarboxylate :
Figure imgf000044_0001
To a stirred solution (S)-2-(methoxycarbonylamino)-4-methylpentanoic acid (step 1 , about 8.0 g, 42.328 mmol) in EtOAc (50 ml) at about 0 °C l -Ethyl-3-(3- Dimethylaminopropyl)carbodiimide (8.925 g, 46.560 mmol), hydroxybenzotriazole (7. 13 g, 49.560 mmol) and NMM (5.6 ml, 50.793 mmol) were added and stirred for 30 minutes then t-Butyl carbazate (6. 152 g, 46.560 mmol) was added and stirred at room temperature for 18 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc and washed with water, saturated NaHC03 solution and brine and the organic layer was concentrated under reduced pressure and the resulting crude was purified by silica gel column ( 100-200 mesh, elution 35% EtOAc in hexane) to afford the title compound as an off white semi solid. Wt: 8.0 g; Yield: 69.2%. Ή NMR (300 MHz, CDCI3): δ 7.96 (brs, 1 H), 6.46 (brs, 1 H), 5.09 (d, 1 H, J = 7.8 Hz), 4.29-4. 18 (m, 1 H), 3.70 (s, 3H), 1 .78- 1 .48 (m, 3H), 1 .46 (s, 9H), 0.96-0.93 (m, 6H); Mass: [M+Na]+ 326 ( 100%); IR (KBr, cm '): 343 1 , 2930, 1728, 1607, 1503, 143 1 , 1251 , 1218, 1 155, 1 130, 1052, 1034, 786,
680.
Step 3: synthesis of (S)-methyl 1 -hydrazinyl-4-methyl- 1 -oxopentan-2-xlcarbamate :
Figure imgf000044_0002
A solution of (S)-tert-butyl 2-(2-(methoxycarbonylamino)-4- methylpentanoyl)hydrazinecarboxylate (step 2, about 1 .0 g, 3.30 mmol) in 2N HC1 and dioxane (20 ml) at about 0°C was stirred and allowed to reach room temperature, then continued stirring for about 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated, basified with solid NaHC0 , extracted with EtOAc and dried over Na2S04 and the solvent was evaporated and the resulting crude was proceeded for next reaction without characterization. Wt: 0.66 g: Yield: 100 %.
Step 4: Synthesis of (S, E)-methyl 4-methyl-l -oxo-l -(2-(4-(pyridin-2-
Figure imgf000044_0003
Figure imgf000045_0001
A solution of (S)-methyl l-hydrazinyl-4-methyl- l-oxopentan-2-ylcarbamate (step 3, about 0.66 g, 3.300 mmol) and 4-(pyridin-2-yl)benzaldehyde (about 0.604 g, 3.300 mmol) in isopropanol and stirred at reflux temperature for about 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated and purified by silica gel column (100-200 mesh, elution 30% EtOAc in hexane) to afford the title compound as a semi solid. Wt: 0.550 g: Yield: 45.83%. Ή NMR (300 MHz, CD3OD): δ 8.63 (d, 1H, J = 4.8 Hz), 8.23 (s, 1H), 8.08-7.81 (m, 6H), 7.41-7.38 (m, 1H), 4.28-4.19 (m, 1H), 3.67 (s, 3H), 1.71-1.56 (m, 3H), 1.10- 0.99 (m, 6H); Mass: [M+l]+ 369 (70%), [M+Na]+391 (100%); DR ( Br, cm"1): 3312, 3228, 2959, 1694, 1669, 1535, 1466, 1436, 1358, 1320, 1242, 1110, 1052, 848, 780; M.R: 185.4 °C -187.9 °C.
Step 5: Synthesis of (S)-methyl 4-methyl-l -oxo-l-(2-(4-(pyridin-2- yl)benzyl)hydrazinyl )pentan-2-ylcarbamate:
Figure imgf000045_0002
To a stirred solution of (S,E)-methyl 4-methyl-l-oxo-l-(2-(4-(pyridin-2- yl)benzylidene)hydrazinyl)pentan-2-ylcarbamate (step 4, about 2.7 g, 7.336 mmol) in EtOH (20 ml), palladium on carbon (10%) catalytic amount was added and stirred under H2 gas atmosphere at room temperature for 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through celite bed and the obtained filtrate was concentrated under reduced pressure. The resulting crude was purified by silica gel column (100-200 mesh, elution 40% EtOAc in hexane) to afford the title compound as a semi solid. Wt: 2.4 g: Yield: 88.8 %; Ή NMR (300 MHz, CD3OD): δ 8.60-8.59 (m, 1H), 7.95-7.79 (m, 4H), 7.49 (d, 2H, J=
8.4 Hz), 7.36-7.29 (m, 1H), 4.10-3.99 (m, 1H), 3.99 (s, 2H), 3.61 (s, 3H), 1.59-1.32 (m, 3H), 0.87-0.83 (m, 6H); Mass: [M+l]+ 371 (20%), [M+Na]+393 (100%); IR (KBr, cm '): 3296, 3059, 2956, 2928, 2869, 2364, 2345, 1674, 1536, 1467, 1437, 1437, 1355, 1253, 1122, 1053, 1017,777. Step 6: Synthesis of (S) methyl -l-(2-((2S,3S)-3-(tert-butoxycarbonylamino)-2- hydrox -4^henylbut}'l)-2-(4-(pyridin-2-yl)benzyl)hydrazinyl)-4-methyl-l- oxopentan-2-ylcarbamate :
Figure imgf000046_0001
A stirred solution of (S)-methyl 4-methyl-l-oxo-l-(2-(4-(pyridin-2- yl)benzyl)hydrazinyl)pentan-2-ylcarbamate (step 5, about 2.4 g, 6.486 mmol) and tert-butyl (S)-l-((R)-oxiran-2-yl)-2-phenylethylcarbamate (about 1.7 g, 6.486 mmol) in Isopropanol (40 ml) was refluxed for about 16 hours. After completion of the reaction (monitored by TLC), the solvent was cooled to room temperature and poured in to an ice water, the resulting solid title compound was filtered and dried under vacuum. Wt:1.5 g; Yield: 44%; Ή NMR (300 MHz, CDCh): δ 8.67 (d, 1H, J = 4.8 Hz).7.93 (d, 2H, J = 8.1 Hz), 7.81-7.68 (m, 2H), 7.41 (d, 2H, J = 7.8 Hz), 7.38-7.11 (m, 6H), 6.80 (s, 1H), 5.10 (d, 1H, J = 9.3 Hz), 4.90-4.70 (m, 3H), 4.12 (d, 1H, J = 13.5 Hz), 3.80-3.78 (m, 2H), 3.77-3.62 (m, 1H), 3.61 (s, 3H), 3.58-3.46 (m, 1H), 2.98-3.86 (m.2H), 2.63-2.52 (m, 1H), 1.37 (s, 9H), 1.30-1.11 (m, 3H), 0.79-
0.64 (m, 6H); Mass: [M+H]+634 (100%); IR (KBr, cm"1): 3373, 3263, 2958, 1691, 1516, 1467, 1366, 1284, 1249, 1171, 1052, 1018, 779, 751; M.R: 176.4 °C -183.1 °C.
Step 7: Synthesis of methyl (S)-l-(2-((2S3S)-3-amino-2-hydroxy-4-phenylb tyl)-2- (4-(pyridin-2-yl)benzyl)hydrazinyl)- -methyl-l-oxopentan-2-ylcarbainate:
Figure imgf000046_0002
To a stirred solution of (S) methyl -l-(2-((2S,3S)-3-(tert- bittoxycarbonylamino)-2-hydrox -4-plienylbut}'l)-2-(4-(pyridin-2- yl)benzyl)hydrazinyl)-4-methyl-l-oxopentan-2-ylcarbamate (Step 6, about 0.4 g,
0.631 mmol) in tetrahydrofuran (8.0 ml) at 0°C, 4N HCl (4 ml) was added drop wise and stirred at about 50 °C for about 3 hours. After completion of the reaction the solvent was removed and dissolved in EtOH, concentrated and dried under vacuum, which was used as such for further reactions. Intermediate 12: Preparation of (R)-methyl l -(2-(4-(2-(4-ethylpiperazin- l -yl) ethoxy) benzyl) hydrazinyl)-3, 3-dimethyl- l -oxobutan-2-ylcarbamate:
Figure imgf000047_0001
Step 1: Synthesis of 4-(2-bromoethoxy) benzaldehyde
Figure imgf000047_0002
To a stirred solution of 4-hydroxy benzaldehyde ( 10 g) in dry acetone ( 100 ml), K2C03 (45.2 g) was added followed by 1 , 2-dibromoethane at room temperature. The reaction mixture was stirred at reflux temperature. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through celite and washed with ethyl acetate. Combined organic layers were concentrated under reduced pressure and purified by column chromatography using hexane and ethyl acetate mixture as an eluent to afford the title compound (6.5 g). H NMR (300 MHZ, CDC13): 9.89 ( 1 H, s), 7.93-7.86 (2H, dd, J= 7.0, 1 .8 Hz), 7.00- 7.03 (2H, dd, J= 7.0, 1 .7 Hz), 4.37 (2H, t, J=6.0 Hz), 3.67 (2H, t, J= 6Hz); Mass: (M- l ); 227.
Step 2: Synthesis of 4-(2-(4-ethylpiperazin-l-yl) ethoxy) benzaldehyde:
Figure imgf000047_0003
To a stirred solution of 4-(24oromoethoxy) benzaldehyde ( 10 g) in dry DMF, n-ethyl piperzine was added at room temperature. The reaction mixture was refluxed for 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was extracted in to ethyl acetate, washed with brine solution and dried over Na2S04 then concentrated under reduced pressure and purified by column chromatography using hexane and ethyl acetate mixture as an eluent to afford 9.2 g of pure product. H NMR (300 MHZ , CDC13 ): 9.89 ( l H, s), 7.93-7.86 (2H, dd, J= 7.0, 1.8 Hz), 7.00-7.03 (2H, dd, J= 7.0, 1 .7 Hz), 4.37 (2H, t, J=6.0 Hz), 3.67 (2H, t, J= 6Hz), 2.77-2.87 (8H, m), 2.61 -2.64 (2H, m), 1 . 16- 1 .21 (m, 3H); Mass: (M- l ); 262. Step 3: Synthesis of (R, Z)-methyl l-(2-(4-(2-(4-ethylpiperazin-l -yl) ethoxy) benzylidene) hydrazinyl)-3, 3-dimethyl-l -oxob tan-2-ylcarbamate:
Figure imgf000048_0001
To a solution of (S)-methyl l -hydrazinyl-3,3-dimethyl- l -oxobutan-2- ylcarbamate (Step 3 for Intermediate 8, 5.35 g) dissolved in EPA ( 100 ml) then 4-(2- (4-ethylpiperazin- l -yl) ethoxy) benzaldehyde (6.9 g) added at ambient temperature.
The reaction mixture was refluxed for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated, diluted with ethyl acetate, washed with water, brine solution and dried over Na2S04 then concentrated under reduced pressure and purified by column chromatography using hexane and ethyl acetate mixture as an eluent to afford the tittle compound (4.0 g) yield 37 %. H
NMR (300 MHZ. CDC13): 7.71 (s, 1 H), 7.22-7.25 (d, J=9Htz, 2H), 685-6.86 (d, J=3Htz, 2H), 5.49-5.52 (m, 1 H), 4.08-4. 1 1 (m, 2H), 3.66-3.89 (m, 3H), 3.64 (s, 3H), 2.84-2.87 ( m, 2H), 2.61 -2.77 (m, 8H), 1 . 16- 1 .21 (m, 3H), 1 .00 (s, 9H); Mass: (M+Na) 447 ( 100 %).
Step 4: Synthesis of (R)-methyl l -(2-(4-(2-(4-ethylpiperazin-l -yl) ethoxy) benzyl) hydrazinyl)-3, 3 -dimethyl- 1 -oxobutan-2-ylcarbamate:
To a stirred solution of (R, Z)-methyl l -(2-(4-(2-(4-ethylpiperazin- l -yl) ethoxy) benzylidene) hydrazinyl)-3, 3-dimethyl- l -oxobutan-2-ylcarbamate (4.0 g) in ethanol (50 ml) Pd/C (50 mg) was added under nitrogen atmosphere. Then the reaction mixture was stirred at room temperature under hydrogen atmosphere for 18 hours. After completion of the reaction (monitored by TLC), filtered through celite and washed with ethyl acetate. Combined organic layers were concentrated under reduced pressure and purified by column chromatography using hexane and ethyl acetate mixture as an eluent to afford 3.0 g of pure product (75 % yield). H NMR
(300 MHZ, CDC13): 7.7 1 (s, 1 H), 7.22-7.25 (d, J=9Htz, 2H), 6.85-6.86 (d, J=3Htz, 2H), 5.49-5.52 (m, 1 H), 4.08-4. 1 1 (m, 2H), 3.66-3.89 (m, 3H), 3.64 (s, 3H), 2.84- 2.87 (m, 2H), 2.61 -2.77 (m, 8H), 1.16- 1.21 (m, 3H), 1 .00 (s, 9H); Mass: (M+Na ) 449( 100 % ). Intermediate 13 : Preparation of (R)-methyl l -(2-(4-(2-(3, 4-dihydroquinolin- l (2H)- yl) ethoxy) benzyl) hydrazinyP-3, 3-dimethyl- l -oxobutan-2-ylcarbamate:
Figure imgf000049_0001
Step 1 : Synthesis of 4-(2-(3, 4-dihydroquinolin- 1 (2H )-yl) ethoxy) benzaldehyde:
Figure imgf000049_0002
To a stirred solution of 4-(2-bromoethoxy) benzaldehyde (Step 1 for
Intermediate 12, 10 g) in dry DMF ( 100 ml) 1 ,2,3,4-tetrahydroquinoline ( 13 g) was added at room temperature. The reaction mixture was refluxed for 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was extracted in to ethyl acetate, washed with brine solution and dried over Na2S04 then concentrated under reduced pressure and purified by column chromatography using hexane and ethyl acetate mixture as an eluent to afford 9.2 gr of pure product. H NMR (300 MHZ, CDC13): 7.96-7.97 (m, 4H), 7.93-7.86 (2H, dd, J= 7.0, 1 .8 Hz), 7.00-7.03 (2H, dd, J= 7.0, 1.7 Hz), 4.37 (2H, t, J=6.0 Hz), 3.67 (2H, t, J= 6Hz); Mass:(M- l ); 280.
Step 2: Synthesis of (R, Z)-methyl l-(2-(4-(2-(3, 4-dihydroquinolin-l(2H)-yl) ethoxy) benzylidene) hydrazinyl)-3, 3 -dimeth - l -oxobutan-2-ylcarbamate:
Figure imgf000049_0003
To a solution of (S)-methyl l -hydrazinyl-3,3-dimethyl- l -oxobutan-2- ylcarbamate (Step 3 for Intermediate 8, 5.35 g) EPA ( 100 ml), 4-(2-(3, 4- dihydroquinolin- l (2H)-yl) ethoxy) benzaldehyde (step 1 , 7.4 g) was added at ambient temperature. The reaction mixture was refluxed for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated, diluted with ethyl acetate, washed with water, brine solution and dried over Na?S04 then concentrated under reduced pressure and purified by column chromatography using hexane and ethyl acetate mixture as an eluent to afford the tittle compound (4.0 g) yield 37 %. H NMR (300 MHZ, CDC13): 7.71 -7.74 (m, 4H), 7.22-7.25 (d, J=9Htz, 2H), 685-6.86 (d, J=3Htz, 2H), 5.49-5.52 (m, 1 H), 4.08- 4. 1 1 (m, 2H), 3.66-3.89 (m, 3H), 3.64 (s, 3H), 2.84-2.87 (m, 2H), 2.61 -2.77 (m, 8H), 1 . 16- 1 .21 (m, 3H), 1 .00 (s, 9H); Mass: (M+ l ) 467 ( 100 %).
Step 3: Synthesis of (R)-methyl l-(2-(4-(2-(3, 4-dihydroquinolin-l(2H)-yl) ethoxy) benzyl) hydrazinyl)-3, 3 -dimethyl- l -oxobutan-2-ylcarbamate:
To a stirred solution of (R, Z)-methyl l -(2-(4-(2-(3, 4-dihydroquinolin- l (2H)-yl) ethoxy) benzylidene) hydrazinyl)-3, 3-dimethyl- l -oxobutan-2- ylcarbamate (Step 2, 4.0 g) in ethanol (50 ml) Pd/C (50 mg) was added under nitrogen atmosphere then the reaction mixture was stirred at room temperature under hydrogen atmosphere for 18 hours. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through celite and washed with ethyl acetate. Combined organic layers were concentrated under reduced pressure and purified by column chromatography using hexane and ethyl acetate mixture as an eluent to afford 3.0 gr of pure product ( 75 % yield). H NMR (300 MHZ, CDC13): 7.71 -7.74 (m, 4H), 7.22-7.25 (d, J=9Htz, 2H ), 685-6.86 (d, J=3Htz, 2H), 5.49-5.52 (m, 1 H), 4.08-4. 1 1 (m, 2H), 3.66-3.89 (m, 3H), 3.64 (s, 3H), 2.84-2.87 (m, 2H), 2.61 -2.77 (m, 8H), 1. 16- 1 .21 (m, 3H), 1 .00 (s, 9H); Mass: (M+Na) 491 ( 100 %).
Intermediate 14: Preparation of (R)-methyl l -(2-(4-(3-(3,4-dihydroquinolin- l (2H)- yl)propoxy)benzyl)hydrazinyl)-3,3-dimethyl- l -oxobutan-2-ylcarbamate:
Figure imgf000050_0001
Step 1: Synthesis of 4-(3-bromopropoxy)benzaldeh
Figure imgf000051_0001
To a stirred solution of 4-hydroxy benzaldehyde ( 10 g) in dry acetone ( 100 ml) K2CO3 (45.2 g) was added followed by 1 ,3-dibromopropane (65.90 g) at room temperature and the reaction mixture was stirred at reflux temperature. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through celite and washed with ethyl acetate, combined organic layers were concentrated under reduced pressure and purified by column chromatography using hexane and ethyl acetate mixture as an eluent to afford 6.5 gr of pure product (35 % yield). H NMR (300 MHZ , CDC13): 9.89 ( 1 H, s), 7.93-7.86 (2H, dd, J= 7.0, 1.8 Hz), 7.00-7.03 (2H, dd, J= 7.0, 1.7 Hz), 4.37 (2H, t, J=6.0 Hz), 3.67 (2H, t, J= 6Hz), 3.40-3.36 (m, 2H); Mass: (M- l ); 240.
Similarly, Step 2 and step 3 procedure are same as in Intermediate 13 to get the Intermediate 14.
Intermediate 15: Preparation of methyl N-[( l S)-2,2-dimethyl- l -[N'-( ( 4-[2- (mo holin-4-yl)ethoxylphenyl ) methyl)hvdrazinecarbonyllpropyllcarbamate:
Figure imgf000051_0002
Step 1 : Synthesis of 4-(2-bromoethox benzaldeh
Figure imgf000051_0003
To a stirred solution of 4-hydroxybenzaldehyde (20 g) in dry acetone (200 ml) K2C03 ( 104 g) was added followed by 1 ,2-dibromoethane at room temperature. The reaction mixture was stirred at reflux temperature overnight. After completion of the reaction (monitored by TLC), the reaction mixture was filtered threw celite and washed with ethyl acetate. Combined organic layers were concentrated under reduced pressure and purified by column chromatography using hexane and ethylacetate mixture as an eiuent to afford 13 gr of pure product (34.7 % yield). Ή NMR (300 MHZ, CDC ): δ 9.89 ( 1 H, s), 7.93-7.86 (2H, dd, J = 7.0, 1 .8 Hz), 7.00- 7.03 (2H, dd, J = 7.0, 1 .7 Hz), 4.37 (2H, t, J = 6.0 Hz), 3.67 1 (2H, t, J = 6Hz); Mass: [Μ- 1 ]' 227 (70%); IR (KBr): 3347, 2798, 2736.9, 1687.4, 1603, 1578.7, 1253,
1230.3, 1 163.3 cm" 1
Step 2: Preparation of 4-(2-morpholinoethoxy)benzaldehyde:
Figure imgf000052_0001
To a stirred solution of 4-(2-bromoethoxy)benzaldehyde (20 g) in dry DMF ( 100 ml) morpholine (30.56 g) was added at room temperature then the reaction mixture was refluxed for over night. After completion of the reaction (monitored by TLC), the reaction mixture was extracted into ethyl acetate, washed with brine solution, dried over Na2S04, and concentrated under reduced pressure to give crude product. The crude compound was purified by column chromatography using hexane and ethyl acetate mixtures as eluent and 100-200 silica-gel mesh as a stationary phase to get 19 gr of pure product (92.2 % Yield). Ή NMR (300 MHZ, CDCh): δ 9.89 ( 1 H, s), 7.85-7.82 (2H, dd, J = 6.9, 2. 1 Hz), 7.00-7.03 (2H, dd, J = 6.9, 2. 1 Hz), 4. 19 (2H, t, J = 5.7 Hz), 3. 74 (4H, t, J = 4.6Hz), 2.8 ( 2H, t, J = 5.7 Hz),
2.59 (2H, t, J = 4.6 Hz); Mass: [M++ H] 236 ( 100%); IR (KBr): 3372, 2956, 2855, 2805, 1689.9, 1602, 1578.4, 1256.0, 123 1 . 1 , 1 161.5 cm 1.
Step 3: Synthesis of methyl N-[(lS)-2,2-diinethyl-]-{N'-[(JE)-(4-[2-(morpholin-4- yljethoxy] phenyl }methylidene]hydrazinecarbonylj propyl] carbamate:
Figure imgf000052_0002
To a solution of (S)-methyl l -hydrazinyl-3,3-dimethyl- l -oxobutan-2- ylcarbamate (Step 3 for Intermediate 8, 5.35 g) dissolved in ΓΡΑ ( 100 ml) benzaldehyde derivative (6. 19 g) at ambient temperature and the reaction mixture was refluxed for over night. After completion of the reaction (monitored by TLC), the reaction mixture was poured in ice cold water to get white solid. The solids were filtered and dissolved in ethyl acetate, dried over Na2S04, concentrated under reduced pressure to give crude product. This crude was further purified by column chromatography using hexane and ethylacetate mixtures as eluent to give 4 gr of pure compound (36.36 % Yield). Ή NMR (300 MHZ, CD OD): δ 8.08 ( 1 H, s), 7.679-7.742 (2H, dd, J = 6.9 & 1.8 Hz), 6.98-7.01 (2H, dd, J = 6.9 & 1.8 Hz), 4.63 (1H, brs), 4.17-4.21 (2H, m), 4.12 (1H, brs), 3.71-3.74 (5H, m), 3.65 (3H, s), 2.83- 2.864 (2H, m), 2.6 (4H, m), 0.99 (9H, s); Mass: [M++H] 421 (100%). Step 4: Synthesis of methyl N-[( lS)-2,2-dimethyl-l -[N'-({4-[2-(morpholin-4- yljethoxy] phenyl }methyl)hydrazinecarbony I] propyl] carbamate:
To a stirred solution of (R,E)-methyl 3,3-dimethyl-l-(2-(4-(2- mo holinoethoxy)benzylidene)hydrazinyl)-l-oxobutan-2-ylcarbamate (Step 3, 4 g) in ethanol (50 ml) Pd/C (10 mg) was added under nitrogen atmosphere. Then the reaction mixture was stirred at room temperature for over night under hydrogen conditions. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through celite and concentrated under reduces pressure to give crude compound. The crude product was then purified by column chromatography using hexane and ethyl acetate mixtures as eluent to afford 3 gr of pure product (75 % yield). Ή NMR (300 MHZ, CDCI3): δ 7.29 (2H, d, J = 8.4 Hz), 6.90 (2H, d, J =
8.4 Hz), 4.11-4.13 (2H, m), 3.83-3.86 (3H, m), 3.7-3.72 (5H, m), 3.65 (3H, s), 0.98 (9H, s); Mass: [M++H] 423 (100%); IR (KBr): 3303, 2960, 2870.6, 1713, 1661, 1533, 1369, 1247, 1117, 1053, 1020, 958, 810, 776 cm"1.
Examples
Example 1: Preparation of (lS,3R)-3-acetamjdo-N-((2S,3S)-4-(2-((lS,3R)-3- acetamido-2,2-dimethylcyclobutanecarbonyl)-l-(4-(pyridin-2-yl)benzyl)hydrazinyl)- 3-hydroxy-l-phenylbutan- -yl)-2,2-dimethylcvclobutanecarboxamide:
Figure imgf000053_0001
To a stirred solution (lS,3R)-3-acetamido-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 1, about 0.330 g, 1.777 mmol) in DMF (8 ml) N,N,N',N'-tetramethyl-o-(benzotriazol-l-yl)uronium tetrafluoroborate (TBTU, about 1.14 g, 3.545 mmol) was added at about 0 °C, after 10 minutes (2S,3S)-3-amino-4-phenyl-l-(l-(4-(pyiidin-2-yl)benzyl)hydrazinyl)butan-2-ol (Intermediate 7, about 0.419 g, 0.888 mmol) in DMF (3 ml) and N-methyl morpholine (0.979 ml, 8.886 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc and washed with water, brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column ( 100-200 mesh, elution 3% MeOH in DCM) to afford the title compound as an off white solid. Wt: 0. 1 g; Yield: 16. 18%; Ή NMR (300 MHz, CD3OD): δ 8.59 (d, 1 H, J =4.2 Hz), 7.89-7.79 (m, 3H), 7.50 (d, 2H, J = 7 Hz), 7.36-7.32 (m, 2H), 7.23-7. 14 (m, 5H), 4.08-3.87 (m, 5H), 2.92-2.76 (m, 3H), 2.61 -2.45 (m, 2H), 2.20- 1.98 (m, 6H), 1. 14 (s, 3H), 1.02 (s, 3H), 0.74 (s, 3H), 0.41 (s, 3H); Mass: [M+ l ]+ 697 (4%), [M+Na]+ 719 ( 100%); HPLC: 93.07%; IR (KBr, cm" 1 ): 3424, 3314, 2927, 1656, 1648, 1510, 1370, 1278, 1 154, 928, 780; M.R: 144.5 °C - 157.3 °C.
Example 2: Preparation of methyl ( l R,3S )-3-((2S,3S)-4-(2-(( l S,3R)-3- methylcarbamato-2,2-dimethylcyclobutanecarbonyl)- l -(4-(pyridin-2- yl)benzyl)hydrazinyr)-3-hydroxy- l -phenylbutan-2-ylcarbamoyl )-2,2- dimethylcyclobutylcarbamate:
Figure imgf000054_0001
thoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 2, about 0.357, 1.776 mmol) in DMF (8 ml) N,N,N',N'-tetramethyl-o-(benzotriazol- l -yl)uronium tetrafluoroborate (TBTU, about 1 . 14 g, 3.545 mmol) was added at about 0 °C, after 10 minutes (2S,3S)-3-amino-4-phenyl- l -( l -(4-(pyridin-2-yl)benzyl)hydrazinyl)butan-2-ol (Intermediate 7, about 0.419 g, 0.888 mmol) in DMF (3 ml) and diisopropyl ethylamine (about 1 .5 ml, 8.886 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc and washed with water, saturated NaHC0 solution, brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column ( 100- 200 mesh, elution 2 % MeOH in DCM) to afford the title compound as an off white solid. Wt: 0. 1 g; Yield: 15.47%; Ή NMR (300 MHz, CDCh): 5 8.68 (d, 1 H, 7 = 4.8 Hz), 7.94 (d, 2H, J = 9.3 Hz), 7.78-7.62 (m, 2H), 7.40 (d, 2H, J = 8. 1 Hz), 7.25-7. 18 (m, 6H), 5.89 (brs, 2H), 5.16-4.98 (m, 2H), 4.88 (brs, 2H), 4.09-3.83 (m, 5H), 3.66 (s, 3H), 3.63 (s, 3H), 3.49-3.31 (m, 1H), 2.91 (m, 2H, J = 7.5 Hz), 2.77 (d, 1H, J = 12 Hz), 2.45-2.16 (m, 3H), 2.06-1.91 (m, 3H), 1.19 (s, 3H), 1.05 (s, 3H), 0.78 (s, 3H),0.51 (s, 3H); Mass: [M+l]+729 (45%), [M+Na)+751 (100%); HPLC: 90.9%; IR (KBr, cm"1): 3424, 3328, 2955, 1706, 1684, 1649, 1653, 1510, 1456, 1354, 1260, 1157, 1054, 1016, 779; M.R: 144.5 °C -157.3 °C.
Example 3: Preparation of ten-butyl (lS,3R>-3-((2S,3S)-4-(2-((lR.3S)-3-(tert- butyloxycarbonylamino)-2,2-dimethylcyclobutanecarbonyl)-l-(4-(pyridin-2- yl)benzyl)hydrazinyl)-3-hydroxy-l-phenylbutan-2-ylcarbamoyl)-2,2- dimethylcyclobutylcarbamat
Figure imgf000055_0001
To a stirred solution of (lS,3R)-3-(tert-butoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 3, about 0.553 g, 2.275 mmol) in DCM (4 ml) hydroxybenzotriazole (about 0.653 g, 4.269 mmol) and l-Ethyl-3-(3-
Dimethylaminopropyl)carbodiimide (about 1.04 g, 5.685 mmol) were added at about 0 °C, after 10 minutes (2S,3S)-3-amino-4-phenyl-l-(l-(4-(pyridin-2- yl)benzyl)hydrazinyl)butan-2-ol (Intermediate 7, about 0.671 g, 1.423 mmol) in DCM (1 ml), DMF (0.5 ml) and diiospropyl ethylamine (about 1.2 ml, 7.115 mmol) were added drop wise and the reaction mass was stirred 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, saturated NaHC03 solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude purified by silica gel column (100-200 mesh, elution 50% EtOAc in hexane) to afford the title compound as a off white solid. Wt: 0.120 g; Yield:
10.43%; Ή NMR (300 MHz, CDC13): δ 8.70 (d, 1H, J = 4.8Hz), 7.95 (d, 2H, J = 8.1Hz), 7.80-7.69 (m, 2H), 7.42 (d, 2H, J = 8.1Hz), 5.90 (d, 1H, J = 8.7 Hz), 5.80 (s, 1H), 4.86 (s, 1H), 4.78 (d, 1H, J = 8.7 Hz), 4.66 (d, 1H, J = 8.7 Hz ), 4.06-3.76 (m, 5H), 3.47 (d, 1H, J = 9.9 Hz), 2.92 (d, 1H, J = 7.8 Hz), 2.83-2.76 (m, lH), 2.49- 2.11 (m, 4H), 2.09-1.88 (m, 3H), 1.44 (s, 9H), 1.43 (s, 9H), 1.21 (s, 3H), 1.07 (s,
3H), 0.81 (s, 3H), 0.51 (s, 3H); Mass: [M+l]+ 813 (4%), [M+Na]+835 (100%); HPLC: 91.7%; I (KBr, cm"1): 3427, 2929, 1703, 1698, 1501, 1366, 1252, 1170, 1045, 778, 747, 702; M.R: 124.9 °C - 136.0 °C.
Example 4: Preparation of benzyl (lS,3R)-3-((2S.3S)-4-(2-((lR,3S)-3- (benzyloxycarbonylamino)-2,2-dimethylcvclobutanecarbonyr)-l-(4-(pyridin-2- yl)benzyl)hydrazinyl)-3-hydroxy-l-phenylbutan-2-ylcarbamoyD-2,2- dimethylcyclobutylcarbama
Figure imgf000056_0001
To a stirred solution (lR,3S)-3-(benzyloxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 4, about 0.671 g, 1.423 mmol) in DCM (4 ml) hydroxybenzotriazole (about 0.653 g, 4.269 mmol) and l-Ethyl-3-(3- Dimethylaminopropyl)carbodiimide (about 1.09 g, 5.691 mmol) were added at about 0 °C, after 10 minutes (2S,3S)-3-amino-4-phenyl-l-(l-(4-(pyridin-2- yl)benzyl)hydrazinyl)butan-2-ol (Intermediate 7, about 0.671 g, 1.423 mmol) in DCM (1 ml), DMF (0.5 ml) and diiospropyl ethylamine (about 1.2 ml, 7.115 mmol) were added drop wise and the reaction mass was stirred 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, saturated NaHC0 solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100-200 mesh, elution 70% EtOAc in hexane) to afford the title compound as an off white solid. Wt: 0.80 g; Yield: 6.6%; Ή NMR (300 MHz, CDC13): δ 8.67 (d, 1H, J =4.2 Hz), 7.93 (d, 2H, J = 8.1 Hz), 7.81-7.68 (m, 2H), 7.42-7.20 (m. 18H), 6.03 (s, 1H), 5.78 (brd, 1H), 5.18-4.91 (m, 7H), 4.14-4.10 (m, 1H), 3.96-(q, 2H, J = 13.5 Hz), 3.84-3.75 (m, 2H), 3.55 (d, 1H, J = 9.9 Hz), 2.90 (d, 1H, J = 7.5 Hz), 2.72-2.45 (m.2H), 2.28-1.95 (m, 7H), 1.16 (s, 3H), 1.06 (s, 3H), 0.59 (s, 3H), 0.56 (s, 3H); Mass: [M+l]+ 882 (35%). [M+Na]+904 (100%); HPLC: 84.9%; IR (KBr, cm"1): 3419, 3315, 3032, 2954, 1702, 1661, 1649, 1513, 1455, 1341, 1257, 1050, 1028, 913, 777, 747, 698; M.R: 107.4 °C -112.4°C. Example 5: Preparation of methyl ( l S R)-3-((2S.3S)-3-hvdroxy-4-(2-(( l R.3S)-3- (methoxycarbonylamino)-2,2-dimethylcyclobutanecarbonyl)- l -(4-(pyridin-2- yPbenzyPhydrazinyl )- 1 -phenylbutan-2-ylcarbamoyl)-2,2- dimethylcyclobutylcarbama
Figure imgf000057_0001
To a stirred solution of ( l R,3S)-3-(methoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 5, about 0.357 g, 1 .776 mmol) in DCM (3 ml) hydroxybenzotriazole (0.408 g, 2.665 mmol) and l -Ethyl-3-(3- Dimethylaminopropyl)carbodiimide (0.681 g, 3.554 mmol) were added at 0 °C, after 10 minutes (2S,3S)-3-amino-4-phenyl- l -( l -(4-(pyridin-2- yl)benzyl)hydraziny])butan-2-ol (Intermediate 7, about 0.419 g, 0.888 mmol) in DCM ( 1 ml), DMF (0.5 ml) and diiospropyl ethylamine (0.76 ml, 4.44 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture w as diluted with DCM and washed with water, saturated NaHCCh solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column ( 100-200 mesh, elution 80% EtOAc in hexane) to afford the title compound as a off white solid. Wt: 120 mg; Yield: 18.5%; Ή NMR (300 MHz, CDC13): δ 8.68 (d, 1 H, J = 4.5 Hz), 7.92 (d, 2H, J = 8. 1 Hz), Ί.Ί9-Ί.63 (m, 2H), 7.39 (d, 2H, J = 7.8 Hz), 7.3 1 -7.09 (m, 6H), 6.09 (s, 1 H), 5.80 (d,
1 H, J = 9 Hz), 5.08-4.86 (m, 3H), 4.14-3.84 (m, 3H), 3.79-3.68 (m, 1 H), 3.63 (s, 3H), 3.61 (s, 3H), 3.59-3.54 (m, 2H), 2.89 (d, 1 H, J = 7.8 Hz), 2.79-2.48 (m, 2H), 2.30-2.85 (m, 7H), 1 . 14 (s, 3H), 1.04 (s, 3H), 0.57 (s, 3H), 0.55 (s,3H); Mass: [M+ l ]+ 729 (30%), [M+Na]+75 1 ( 100%); HPLC: 87.6%; IR (KBr, cm 1 ): 3418, 3301 , 2954, 2868, 1704, 1650, 1644, 15 13, 1460, 1435, 1366, 1350, 1292, 1 257,
1067, 934, 778; M.R: 129.5 °C - 138.9 °C.
Example 6: Preparation of methyl ( l R,3S)-3-((2S,3S)-3-hydroxy-4-(2-((S)-2-
(methoxycarbonylamino)-3,3-dimethylbutanoyl)- l -(4-(pyridin-2- yObenzyPhydrazinyl )- l -phenylbutan-2-ylcarbamoyl)-2.2- dimethylcyclobutylcarbamate:
Figure imgf000058_0001
To a stirred solution of (lS,3R)-3-(methoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 2, about 0.152 g, 0.759 mmol) in DCM (4 ml) at about 0 °C, hydroxybenzotnazole (about 0.145 g, 0.948 mmol) and l-Ethyl-3-(3-Dimethy]aminopropyl)carbodiimide (about 0.181 g, 0.948 mmol) were added, after 10 minutes methyl (S)-l-(2-((2S,3S)-3-amino-2-hydroxy-4- phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2- ylcarbamate (Intermediate 8, about 0.337 g, 0.632 mmol) in DCM (1 ml), DMF (1 ml) and NMM (about 0.34 ml, 3.160 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 48 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHC03 solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100-200 mesh, elution 80% EtOAc in hexane) to afford the title compound as an off white solid. Wt: 0.180 g; Yield: 39.8%; Ή NMR (300 MHz, CDC1?): δ
8.68 (d, 1H), 7.94 (d, 2H, J = 8.1 Hz), 7.80-7.68 (m, 2H), 7.41 (d, 1H, J = 8.4 Hz), 7.32-7.11 (m, 6H), 6.64 (s, 1H), 5.82 (d, 1H, J = 9 Hz), 5.25 (d, 1H, J = 8.4 Hz), 5.08 (d, 1H, J = 9.3 Hz), 4.85 (s, 1H), 4.08-3.88 (m, 3H), 3.84-3.71 (m, 1H), 3.65 (s, 3H), 3.64 (s, 3H), 3.59-3.51 (m, 3H), 2.91 (d, 2H, J = 7.5 Hz), 2.81-2.69 (m, 1H), 2.58-2.48 (m, 1H), 2.31-2.18 (m, 2H), 1.19-1.86 (m, 1H), 1.14 (s, 3H), 0.78 (s, 9H),
0.72 (s, 3H); Mass: [M+l]+717 (15%), [M+Na]+ 739 (100%); HPLC: 98.07%; IR (KBr, cm1): 3433, 3364, 3220, 2952, 1725, 1709, 1675, 1656, 1470, 1241, 1057, 989, 784; M.R: 202.3 °C -212.0 °C. Example 7: Preparation of methyl (S)-l-(2-((2S.3S)-3-((lS,3R)-3-(tert- butoxYcarbonylamino)-2,2-dimethylcvclobutanecarboxamido)-2-hvdiOxy-4- phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2- ylcarbamate:
Figure imgf000059_0001
To a stirred solution of (lS,3R)-3-(tert-butoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 3, about 0.168 g, 0.664 mmol) in DMF (4 ml) hydroxybenzotriazole (about 0.127 g, 0.830 mmol) and l-Ethyl-3-(3- Dimethylaminopropyl)carbodiimide (about 0.159 g, 0.830 mmol) were added at about 0 °C and stirred for about 10 minutes then methyl (S)-l-(2-((2S,3S)-3-amino- 2-hydroxy-4-phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hydrazinyl)-3,3-dimethyl-l- oxobutan-2-ylcarbamate (Intermediate 8, about 0.295 g, 0.553 mmol) in DCM (1 ml), DMF (1 ml) and NMM (about 0.3 ml, 2.768 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 48 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHCC solution and brine. The organic layer was concentrated under reduced pressure and the resulting crude was purified by silica gel column chromatography (100-200 mesh, elution 35% EtOAC in hexane) to afford the title compound as a white solid. Wt: 0.300 g; Yield: 71.6%; Ή NMR (300 MHz, CDC13): δ 8.69 (d, 1H, J = 4.5 Hz), 7.94 (d, 2H, J = 8.1 Hz), 7.81-7.67 (m, 2H), 7.41 (d, 2H, J = 8.1 Hz), 7.34-7.12 (m, 6H), 6.64 (s, 1H), 5.83 (d, 1H, J = 7.5 Hz), 5.25 (d, 1H, J = 8.7 Hz), 4.84 (s, 2H), 4.09-3.86 (m, 3H), 3.80-3.51 (m, 6H), 2.91 (d, 2H, J = 7.5 Hz), 2.85-2.71 (m, 1H), 2.56-2.48 (m, 1H), 2.31-2.12 (m, 2H), 1.98-1.84 (m, 1H), 1.42 (s, 9H), 1.15 (s, 3H), 0.78 (s, 9H), 0.73 (s, 3H); Mass: [M+l]+ 759 (15%), [M+Na]+ 781 (100%); HPLC: 94.3%; ER ( Br, cm"1): 3334, 2965, 1708, 1683, 1679, 1523, 1467.1365, 1251, 1168, 1052, 1019, 894, 779, 708; M.R: 153.6 °C -161.7 °C.
Example 8: Preparation of methyl (S)-l-(2-((2S,3S)-3-((lS,3R)-3-acetamido-2,2- dimethylcvclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-(4-(pyridin-2- yl)benzyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate:
Figure imgf000059_0002
To a stirred solution of (lS,3R)-3-acetamido-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 1, about 0.122 g, 0.664 mmol) in DMF (4 ml) hydroxybenzotriazole (about 0.127 g, 0.830 mmol), l-Ethyl-3-(3- Dimethy]aminopropyl)carbodiimide (about 0.159 g, 0.830 mmol) were added at about 0°C and stirred for about 10 minutes then methyl (S)-l-(2-((2S,3S)-3-amino-2- hydroxy-4-phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hydrazinyl)-3,3-dimethyl-l- oxobutan-2-ylcarbamate (Intermediate 8, about 0.295 g, 0.553 mmol) in DCM (1 ml), DMF (1 ml) and NMM (0.3 ml, 2.768 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 48 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHCC solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100-200 mesh, elution 90% EtOAc in hexane) to afford the title compound as an off white solid. Wt: 0.280 g; Yield: 72%; Ή NMR (300 MHz, CDC13): δ 8.68 (d, 1H, J = 4.2 Hz), 7.93 (d, 2H, J = 8.1 Hz), 7.80-7.68 (m, 2H), 7.42 (d, 2H), 7.34-
7.12 (m, 6H), 6.91 (s, 1H), 6.13 (d, 1H, J = 8.4 Hz), 5.91 (d, 1H, J = 9.3 Hz), 5.29 (d, 1H, J = 7.8 Hz), 4.90 (brs, 1H), 4.09-3.88 (m, 4H), 3.65 (s, 3H), 3.64-3.50 (m, 2H), 2.98-2.84 (m, 2H), 2.81-2.78 (m, 1H), 2.56-2.44 (m, 1H), 2.33-2.14 (m, 2H), 1.96 (s, 3H), 1.95-1.81 (m, 1H), 1.16 (s, 3H), 0.78 (s, 9H), 0.71 (s, 3H); Mass:
[M+l]+ 701 (12%), [M+Na]+ 723 (100%); HPLC: 94.2%; IR (KBr, cm"1): 3347,
2958, 1716.1652, 1516, 1467, 1369, 1251, 1112, 1062, 1017, 780, 748; M.R: 119.7 °C -128.9 °C.
Example 9: Preparation of methyl (S)-l-(2-((2S,3S)-2-hydroxy-3-((lR,3S)-3- (methoxycarbonylamino)-2,2-dimethylcyclobutanecarboxamido)-4-phenylbutyl)-2- (4-(pyridin-2-yl)benzyl)hvdrazinyr)-3,3-dimeth l-l-oxobutan-2-ylcarbamate:
Figure imgf000060_0001
To a stirred solution of (lR,3S)-3-(methoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 5, about 0.114 g, 0.568 mmol) in DCM (4 ml) hydroxybenzotriazole (about 0.108 g, 0.710 mmol) and l-Ethyl-3-(3-
Dimethylaminopropyl)carbodiimide (0.136 g, 0.710 mmol) were added at about 0 °C for about 10 minutes then methyl (S)-l-(2-((2S,3S)-3-amino-2-hydroxy-4- phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2- ylcarbamate (Intermediate 8, about 0.252 g, 0.473 mmol) in DCM (1 ml), DMF (1 ml) and NMM (about 0.26 ml, 2.369 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 48 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHC03 solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100-200 mesh, elution 80% EtOAC in hexane) to afford the title compound as an off white solid. Wt: 0.080 g; Yield: 23.6%. Ή NMR (300 MHz, CDC13): δ 8.70 (d, 1H, J = 4.2 Hz), 7.94 (d, 2H, J = 8.1 Hz), 7.81-7.69 (m, 2H), 7.45 (d, 2H, J = 8.1 Hz), 7.30-7.13 (m, 6H), 6.98 (brs, 1H), 5.78 (d, 1H, J = 9.3 Hz), 5.28 (d, 1H, J = 9.3 Hz), 5.12 (d, 1H, 7 = 8.4 Hz), 4.89 (s, 1H), 4.26-4.14 (m, 1H), 4.07 (d, 1H,J = 13.8 Hz), 3.94 (d, 1H, J = 13.8 Hz), 3.80-3.52 (m, 3H), 3.64 (s, 3H), 3.63 (s, 3H), 2.98-2.89 (m.2H), 2.80-2.67 (m, 2H), 2.29-2.11 (m, 2H), 2.10-1.99 (m, 1H), 1.11 (s, 3H), 0.76 (s, 9H), 0.43 (s, 3H); Mass: [M+l]+ 717 (67%), [M+Na]+ 739 (100%); HPLC: 94.06%; IR ( Br, cm"1): 3425, 2957, 1708, 1675, 1517, 1457, 1368, 1259, 1133, 1058, 1017, 904, 780, 702; M.R: 115.9-121.6 °C.
Example 10: Preparation of methyl (S)-l-(2-((2S,3S)-3-((lR,3S)-3-(tert- butoxycarbonylamino)-2,2-dimethylcvclobutanecarboxamjdo)-2-hydroxy-4- phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hvdrazinyl )-3,3-dimethyl- 1 -oxobutan-2- ylcarbamate:
Figure imgf000061_0001
To a stirred solution of (lR,3S)-3-(tei1-butoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 6, about 0.162 g, 0.663 mmol) in DCM (10 ml), hydroxybenzotriazole (about 0.127 g, 0.829 mmol) and l-Ethyl-3-(3- Dimethylaminopropyl)carbodiimide (about 0.158 g, 0.829 mmol) were added at about 0 °C and stirred about 10 minutes then methyl (S)-l-(2-((2S,3S)-3-amino-2- hydroxy-4-phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hydrazinyl)-3,3-dimethyl-l- oxobutan-2-ylcarbamate (Intermediate 8, about 0.294 g, 0.552 mmol) in DCM (5 ml), DMF (2 ml) and NMM (about 0.3 ml, 2.764 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 48 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHC03 solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100-200 mesh, elution 45% EtOAc in hexane) to afford the title compound as an off white solid. Wt: 0.050 g; Yield: 11.9%; Ή NMR (300 MHz, CDC13): δ 8.68 (d, 1H, J = 4.5 Hz), 7.92 (d, 2H, J = 8.4 Hz), 7.81-7.66 (m, 2H), 7.43 (d, 2H, J - 8.4 Hz), 7.32-7.11 (m, 7H), 5.81 (d, 1H, J = 8.7 Hz), 5.30 (d, 1H, J = 8.7 Hz), 4.94 (d, 1H, J = 8.4 Hz), 4.27-4.18 (m, 1H), 4.04 (d, 1H, J =
13.8 Hz), 3.91 (d, 1H, J = 13.5 Hz), 3.79-3.50 (m, 3H), 3.62 (s, 3H), 2.98-2.83 (m, 2H), 2.79-2.63 (m, 2H), 2.30-1.99 (m, 4H), 1.40 (s, 9H), 1.10 (s, 3H), 0.74 (s, 9H), 0.40 (s, 3H); Mass: [M+l]+ 759 (50%), [M+Na]+ 781 (100%); HPLC: 92.1%; IR (KBr, cm"1): 3354, 2959, 1710, 1679, 1649, 1653, 1588, 1561, 1513, 1467, 1456, 1437, 1366, 1253, 1171, 1109, 1059, 1019, 916, 874, 827, 779, 748, 700; M.R:
116.3 °C -120.9 °C.
Example 11: Preparation of methyl (S)-l-(2-((2S,3S)-3-((lR,3S)-3- (benzyloxycarbonylamino)-2,2-dimethylcyclobutanecarboxamido)-2-hydroxy-4- phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hvdrazinyr)-3,3-dimethyl-l-oxobutan-2- ylcarbamate:
Figure imgf000062_0001
To a stirred solution of (l ,3S)-3-(benzyloxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 4, about 0.210 g, 0.758 mmol) in DCM (20 ml), hydroxybenzotriazole (about 0.145 g, 0.947 mmol) and l-Ethyl-3-(3-
Dimethylaminopropyl)carbodiimide (about 0.181 g, 0.947 mmol) were added at about 0 °C and stirred for about 10 minutes then methyl (S)-l-(2-((2S,3S)-3-amino- 2-hydiOxy-4-phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hydrazinyl)-3,3-dimethyl-l- oxobutan-2-ylcarbamate (Intermediate 8, about 0.336 g, 0.631 mmol) in DCM (20 ml), DMF (1 ml) and NMM (0.35 ml, 3.159 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 48 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHC03 solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100-200 mesh, elution 45% EtOAC in hexane) to afford the title compound as an off white solid. Wt: 0.050 g; Yield: 10%; Ή NMR (300 MHz, CDC13): δ 8.68 (d, 1H, J = 4.2 Hz), 7.93 (d, 2H, J = 7.8 Hz), 7.79-7.68 (m, 2H), 7.43 (d, 2H, J = 7.8 Hz), 7.39-7.10 (m, 11H), 6.65 (s, 1H), 5.69 (d, 1H, J = 9.0 Hz), 5.23 (d, 1H, J = 8.7 Hz), 5.16-4.98 (m, 4H), 4.84 (s, 1H), 4.21-4.11 (m, 1H), 4.06 (d, 1H, J = 13.5 Hz), 3.91 (d, 1H, J = 14.1 Hz), 3.83-3.74 (m, 1H), 3.64 (s, 3H), 3.62-3.55 (m, 3H), 2.90
(d, 2H, J = 7.5 Hz), 2.79-2.59 (m, 2H), 2.30-1.98 (m, 3H), 1.11 (s, 3H), 0.74 (s, 3H), 0.43 (s, 3H); Mass: [M+l]+793 (60%), [M+Na]+815 (100%); HPLC: 99.12%; IR ( Br, cm"1): 3419, 3172, 3031, 2958, 1671, 1534, 1467, 1369, 1305, 1253, 1221, 1109, 1054, 828,778, 747; M.R: 112 °C -117.1 °C.
Example 12: Preparation of methyl (S)-l-(2-benzyl-2-((2S,3S)-2-hydroxy-3- (( lS,3R)-3-(methoxycarbonylarnino)-2,2-dimethylcyclobutanecarboxamido)-4- phenylbutyl)hvdrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate:
Figure imgf000063_0001
To a stirred solution (lS,3R)-3-(methoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 2, about 0.216 g, 1.077 mmol) in DCM (6 ml), hydroxybenzotriazole (about 0.206 g, 1.347 mmol) and l-Ethyl-3-(3- DimethylaminopiOpyl)carbodiimide (about 0.258 g, 1.347 mmol) were added at about 0 °C and stirred for about 10 minutes then methyl (S)-l-(2-((2S,3S)-3-amino- 2-hydroxy-4-phenylbutyl)-2-benzylhydrazinyl)-3,3-dimethyl- 1 -oxobiitan-2- ylcarbamate (Intermediate 9, about 0.409 g, 0.898 mmol) in DCM (4 ml), DMF (1 ml) and NMM (about 0.5 ml, 4.49 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 48 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHC03 solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100-200 mesh, elution 55% EtOAc in hexane) to afford the title compound as a white solid. Wt: 0.3 g; Yield: 52.2%; Ή NMR (300 MHz, CDC13): δ 7.37-7.12 (m, 10H), 6.55 (s, 1H), 5.85 (d, 1H, J = 9 Hz), 5.26 (d, 1H, J = 9 Hz), 5.06 (d, 1H, J = 9.3 Hz), 4.84 (brs, 1H), 4.08-3.76 (m, 4H), 3.67, 3.65 (2s, 6H), 3.60-3.48 (m, 2H), 2.90 (d, 2H, J = 7.5 Hz), 2.78-2.69 (m, 1H), 2.54-2.42 (m, 1H), 2.32-2.18 (m, 2H), 1.99-1.88 (m, 1H), 1.17 (s, 3H), 0.77 (s, 9H), 0.72 (s, 3H); Mass: [M+l]+640 (95%),
[M+Na]+662 (100%); HPLC: 95.8%; IR ( Br, cm1): 3426, 3351, 3241, 2956, 1703, 1656, 1515, 1455, 1368, 1357, 1256, 1059, 923, 776, 699; M.R: 168.4 °C -172.1 °C.
Example 13: Preparation of methyl (S)-l-(2-((2S,3S)-3-((lS,3R)-3-acetamido-2.2- dimethylcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-benzylhvdrazinyl)-
Figure imgf000064_0001
To a stirred solution of (lS,3R)-3-acetamido-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 1, about 0.199 g, 1.077 mmol) in DMF (4 ml), hydroxybenzotriazole (about 0.206 g, 1.347 mmol) and l-Ethyl-3-(3-
Dimethylaminopropyl)carbodiirrude (about 0.258 g, 1.347 mmol) were added at about 0 °C for about 10 minutes then methyl (S)-l-(2-((2S,3S)-3-amino-2-hydroxy- 4-phenylbutyl)-2-benzylhydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate (Intermediate 9, about 0.409 g, 0.898 mmol) in DCM (1 ml), DMF (1 ml) and NMM (about 0.5 ml, 4.49 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 42 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHCCh solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100-200 mesh, elution 90% EtOAc in hexane) to afford the title compound as a white solid.
Wt: 0.200 g; Yield: 35.6%; Ή NMR (300 MHz, CDC1?): δ 7.32-7.13 (m, 10H), 6.61 (s, 1H), 6.05 (d, 1H, J = 8.7 Hz), 5.90 (d, 1H, J = 9 Hz), 5.26 (d, 1H, J = 8.7 Hz), 4.85 (brs, 1H), 4.08-3.82 (m, 4H), 3.67 (s, 3H), 3.60-3.49 (m, 2H), 2.95-2.86 (m, 2H), 2.79-2.68 (m, 1H), 2.52-2.43 (m, 1H), 2.34-2.22 (m, 2H), 1.98 (s, 3H), 1.97- 1.88 (m, 1H), 1.20 (s, 3H), 0.77 (s, 9H), 0.72 (s, 3H); Mass: [M+l]+ 624 (10%),
[M+Na]+646 (100%); HPLC: 93.2%; IR (KBr, cm"1): 3424, 3350, 2958, 1712, 1648, 1656.1521, 1456, 1370, 1254, 1059, 1020, 745; M.R: 100.3 °C - 148.9 °C. Example 14: Preparation of methyl (S)-l-(2-benzyl-2-((2S,3S)-3-((lS.3R)-3-(tert- butoxycarbonylamino)-2,2-dimethylcyclobutanecarboxamido)-2-hydroxy-4- phenylbutyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate:
Figure imgf000065_0001
To a stirred solution of (lS,3R)-3-(tert-butoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 3, about 0.261, 1.077 mmol) in DCM (6 ml), hydroxybenzotriazole (about 0.206 g, 1.347 mmol) and l-Ethyl-3-(3- Dimethylaminopropyl)carbodiimide (about 0.258 g, 1.347 mmol) were added at about 0 °C and stirred for about 10 minutes then methyl (S)-l-(2-((2S,3S)-3-amino-
2-hydroxy-4-phenylbutyl)-2-benzylhydrazinyl)-3,3-dimethyl-l-oxobutan-2- ylcarbamate (Intermediate 9, about 0.475 g, 0.898 mmol) in DCM (4 ml), DM (1 ml) and NMM (about 0.5 ml, 4.49 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 48 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHC03 solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100-200 mesh, elution 40% EtOAc in hexane) to afford the title compound as an off white solid. Wt: 0.3 g; Yield: 49%; Ή NMR (300 MHz, CDC13): δ 7.37- 7.12 (m, 10H), 6.55 (s, 1H), 5.82 (d, 1H), 5.25 (d, 1H, J = 9 Hz), 4.87-4.78 (m, 2H),
4.02-4.71 (m, 4H), 3.67 (s, 3H), 4.59-4.48 (m, 2H), 2.90 (d, 2H, J = 7.5 Hz), 2.78- 2.70 (m, 1H), 2.51-2.43 (m, 1H), 2.31-2.14 (m, 2H), 1.98-1.86 (m, 1H), 1.43 (s, 9H), 1.18 (s, 3H), 0.77 (s, 9H), 0.73 (s, 3H); Mass: [M+l]+ 682 (10%), [M+Na]+ 704 (100%); HPLC: 99.2%; IR ( Br, cm"1): 3426, 3356, 2962, 1709, 1691, 1514, 1509, 1456, 1367, 1253, 1171, 1053, 1022, 782, 748, 700; M.R: 170.8 °C -174.5°C.
Example 15: Preparation of methyl (S)-l-(2-((2S.3S)-3-((lS.3R)-3-acetamido-2,2- dimethylcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyr)-2-(biphenyl-4- ylmethyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate:
Figure imgf000066_0001
To a stirred solution of ( l S,3R)-3-acetamido-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 1 , about 0. 122 g, 0.569 mrnol) in DMF (6 ml), hydroxybenzotriazole (about 0. 109 g, 0.7 12 mrnol) and l -Ethyl-3-(3- Dimethylaminopropyl)carbodiimide (about 0. 136 g, 0.7 12 mrnol) were added at about 0 °C and stirred for about 10 minutes then methyl (S)- l -(2-((2S,3S)-3-amino- 2-hydroxy-4-phenylbutyl)-2-(biphenyl-4-ylmethyl)hydrazinyl)-3,3-dimethyl- l - oxosbutan-2-ylcarbamate (Intermediate 10, about 0.252 g, 0.474 mrnol) in DCM (5 ml), DMF (2 ml) and NMM (about 0.26 ml, 2.373 mrnol) were added drop wise and the reaction mass was stirred at room temperature for about 24 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHC03 solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column ( 100-200 mesh, elution 90% EtOAc in hexane) to afford the title compound as an off white solid. Wt: 0.150 g; Yield: 45.3%; Ή NMR
(300 MHz, CDC13): δ 7.59-7. 12 (m, 14H), 6.70 (s, l H), 6.08 (d, 1 H, J = 8.7 Hz), 5.91 (d, 1 H, J = 9.3 Hz), 5.27 (d, 1 H, J = 9.0 Hz), 4.88 (s, 1 H), 4. 1 1 -3.82 (m, 4H), 3.65 (s, 3H), 3.65-3.49 (m, 2H), 2.96-2.88 (m, 2H), 2.81 -2.72 (m, 1 H), 2.59-2.48 (m, 1 H), 2.39-2.20 (m, 2H), 1 .96 (s, 3H), 1 .96- 1.83 (m, 1 H), 1.20 (s, 3H), 0.76, 0.74 (2s, 12H); Mass: [M+Na]+ 722 ( 100%); HPLC: 94.4%; IR (KBr, cm" 1 ): 3419, 3063,
2960, 1712, 1652, 1648, 1516, 1456, 137 1 , 1264, 1236, 1074, 1055, 914, 762; M.R: 120.5 °C - 123.4 °C.
Example 16: Preparation of methyl (S)- l -(2-(biphenyl-4-ylmethyl)-2-((2S,3S)-2- hydroxy-3-(( 1 S,3R)-3-(methoxycarbonylamino)-2,2- dimethylcyclobutanecarboxamido)-4-phenylbutyl)hydrazin yl )-3,3-dimethyl- 1 - oxobutan-2-ylcarbamate:
Figure imgf000066_0002
To a stirred solution of (lS,3R)-3-(methoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 2, about 0.114 g, 0.569 mmol) in DCM (15 ml), hydroxybenzotriazole (about 0.109 g, 0.712 mmol) and l-Ethyl-3-(3- Dimethylaminopropyl)carbodiimide (about 0.136 g, 0.712 mmol) were added at about 0 °C and stirred for about 10 minutes then methyl (S)-l-(2-((2S,3S)-3-amino-
2-hydroxy-4-phenylbutyl)-2-(biphenyl-4-ylmethyl)hydrazinyl)-3,3-dimethyl-l- oxosbutan-2-ylcarbamate (Intermediate 10, about 0.252 g, 0.474 mmol) in DCM (5 ml), DMF (2 ml) and NMM (0.26 ml, 2.373 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 24 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHC03 solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100-200 mesh, elution 50% EtOAC in hexane) to afford the title compound as an off white solid. Wt: 0.180 g; Yield: 53%; Ή NMR (300 MHz, CDC13): δ 7.60- 7.11 (m, 14H), 6.74 (s, 1H), 5.88 (d, 1H), 5.28 (d.1H, J= 11.7 Hz), 5.15 (d, 1H,7 =
8.7 Hz), 4.88 (s, 1H), 4.10-3.75 (m, 4H), 3.64 (s, 6H), 3.63-3.51 (m, 2H), 2.92 (d, 1H, J = 7.5 Hz), 2.81-2.70 (m, 1H), 2.59-2.48 (m, 1H), 2.32-2.14 (m, 2H), 1.99-1.87 (m, 1H), 1.17 (s, 3H), 0.76 (brs, 12H); Mass: [M+Na]+ 738 (100%); HPLC: 95.2%; IR (KBr, cm"1): 3434, 3366, 3030, 2953, 1714.1677, 1659, 1513, 1456, 1352, 1245, 1057, 824, 764, 702; M.R: 189.5 °C -193.5 °C.
Example 17: Preparation of methyl (S)-l-(2-(biphenyl-4-ylmethyl)-2-((2S,3S)-3-
((lS.3R)-3-(tert-butoxycarbonylamino)-2,2-dimethylcyclobutanecarboxamido)-2- hydroxy-4-phenylbutyl)hydrazinyr)-3,3-dimethyl-l-oxobutan-2-ylcarbamate:
Figure imgf000067_0001
To a stirred solution of (lS,3R)-3-(tert-butoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 3, about 0.186 g, 0.759 mmol) in DCM (10 ml), hydroxybenzotriazole (about 0.145 g, 0.949 mmol) and l-Ethyl-3-(3- Dimethylarrunopropy carbodiimide (about 0.181 g, 0.949 mmol) were added at about 0 °C and stirred for about 10 minutes then methyl (S)-l-(2-((2S,3S)-3-amino-
2-hydroxy-4-phenylbutyl)-2-(biphenyl-4-ylmethyl)hydrazinyl)-3,3-dimethyl-l- oxosbutan-2-ylcarbamate (Intermediate 10, about 0.336 g, 0.632 mmol) in DCM (5 ml), DMF (2 ml) and NMM (0.35 ml, 3.164 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 24 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHC03 solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100-200 mesh, elution 50% EtOAC in hexane) to afford the title compound as a white solid. Wt: 0.200 g; Yield: 41.6%; Ή NMR (300 MHz, CDCh): δ 7.60- 7.11 (m, 14H), 6.79 (s, 1H), 5.88 (d, 1H, J = 9.0 Hz), 5.28 (d, 1H, J = 9.6 Hz), 4.97- 4.82 (m, 2H), 4.11-3.82 (m, 2H), 3.80-3.70 (m, 1H), 3.70-3.51 (m, 5H), 2.97-2.88
(m, 2H), 2.87-2.72 (m, 1H), 2.59-2.48 (m, 1H), 2.37-2.11 (m, 2H), 2.01-1.87 (m, 1H), 1.42 (s,9H), 1.18 (s, 3H), 0.76 (bis, 12H); Mass: [M+Na]+ 780 (100%); HPLC: 97.1%; IR (KBr, cm"1): 3353, 3060, 2959, 1710, 1687, 1678, 1510, 1501, 1456, 1367, 1259, 1172, 1054, 1021, 884, 762; M.R: 178.4 °C -179.3 °C.
Example 18: Preparation of methyl (S)-l-(2-(biphenyl-4-ylmethyl)-2-((2S,3S)-2- hydroxy-3-(( 1 R,3S)-3-(methoxycarbonylamino)-2,2- dimethylcyclobutanecarboxamido)-4-phenylbutyl)hydrazinyl)-3,3-dimethyl-l- oxobutan-2-ylcarbamate:
Figure imgf000068_0001
To a stirred solution of (lR,3S)-3-(methoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 5, about 0.152 g, 0.759 mmol) in DCM (10 ml), hydroxybenzotriazole (about 0.145 g, 0.949 mmol) and l-Ethyl-3-(3- Dimethylaminopropyl)carbodiimide (about 0.182 g, 0.949 mmol) were added at about 0 °C and stirred for about 10 minutes then methyl (S)-l-(2-((2S,3S)-3-amino-
2-hydroxy-4-phenylbutyl)-2-(biphenyl-4-ylmethyl)hydrazinyl)-3,3-dimethyl-l- oxosbutan-2-ylcarbamate (Intermediate 10, about 0.336 g, 0.632 mmol) in DCM (5 ml), DMF (2 ml) and NMM (about 0.35 ml, 3.164 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 24 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 Na SC>4. The organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100-200 mesh, elution 55% EtOAC in hexane) to afford the title compound as an off white solid. Wt: 0.150 g; Yield: 33.2%; 'HNMR (300 MHz, CDC13): 57.60-7.11 (m, 15H), 5.81 (d, 1H,7 = 8.7 Hz), 5.29 (d, 1H, J = 9.3 Hz), 5.14 (d, 1H, J = 8.4 Hz), 4.93 (s, 1H), 4.27-4.18 (m, 1H),
4.04 (d, 1H, J = 13.5 Hz), 3.89 (d, 1H, J = 13.8 Hz), 3.81-3.77 (m, 1H), 3.71-3.52 (m, 8H), 2.98-2.84 (m, 2H), 2.79-2.69 (m, 2H), 2.31-2.03 (m, 3H), 1.10 (s, 3H), 0.73 (s, 9H), 0.39 (s, 3H); Mass: [M+Na]+ 738 (100%); HPLC: 92.6%; IR (KBr, cm"1): 3419, 3334, 3029, 2957, 1709, 1676, 1516, 1367, 1256, 1194, 1109, 1057, 1020, 825, 763, 699, 565; M.R: 125.7 °C -146.7 °C.
Example 19: Preparation of methyl (S)-l-(2-(biphenyl-4-ylmethyl)-2-((2S,3S)-3-
((lR,3S)-3-(tert-butoxycarbonylamino)-2,2-dimethylcyclobutanecarboxamido)-2- hvdroxy-4-phenylbutyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate:
Figure imgf000069_0001
To a stirred solution of (lR,3S)-3-(tert-butoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 6, about 0.186 g, 0.759 mmol) in DCM (10 ml), hydroxybenzotriazole (about 0.145 g, 0.949 mmol) and l-Ethyl-3-(3- Dimethylaminopropyl)carbodiimide (about 0.181 g, 0.949 mmol) were added at about 0 °C and stirred for about 10 minutes then methyl (S)-l-(2-((2S,3S)-3-amino-
2-hydroxy-4-phenylbutyl)-2-(biphenyl-4-ylmethyl)hydrazinyl)-3,3-dimethyl-l- oxosbutan-2-ylcarbamate (Intermediate 10, about 0.336 g, 0.632 mmol) in DCM (5 ml), DMF (2 ml) and NMM (0.35 ml, 3.164 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 24 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHCCh solution, brine and dried over Na2SOj, the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100-200 mesh, elution 40% EtOAC in hexane) to afford the title compound as an off white solid. Wt: 0.180 g; Yield: 37.5%; Ή NMR (300 MHz, CDCb): 57.58-7.11 (m.15H), 5.79 (d.1H, J = 9.0 Hz), 5.28 (d, 1H, J =
9.3 Hz), 4.98-4.79 (m, 2H), 4.28-4.16 (m, 1H), 4.05 (d, 1H, J = 13.8 Hz), 3.88 (d, 1H, J = 13.8 Hz), 3.80-3.51 (m, 5H), 2.98-3.87 (m, 2H), 2.79-2.69 (m, 1H), 2.30- 2.02 (m, 3H), 1.40 (s, 9H), 1.11 (s, 3H), 0.72 (s, 9H), 0.40 (s, 3H); Mass: [M+l]+ 758 (20%), [M+Na]+780 (100%); HPLC: 97.3%; IR (KBr, cm"1): 3364, 2959, 2343, 1710, 1654, 1509, 1456, 1366, 1253, 1171, 1056, 1021, 914, 762, 699;M.R: 106.6 °C-115.6°C.
Example 20: Preparation of methyl (S)-l-(2-((2S,3S)-3-((lR,3S)-3- (benzyloxycarbonylamino)-2,2-dimethylcyclobutanecarboxamido)-2-hydroxy-4- phenylbutyl)-2-(biphenyl-4-ylmethvDhvdrazinyl)-3,3-dimethyl-l-oxobutan-2- ylcarbamate:
Figure imgf000070_0001
To a stirred solution of ( lR,3S)-3-(benzyloxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 4, about 0.210 g, 0.759 mmol) in DCM (10 ml), hydroxybenzotriazole (about 0.145 g, 0.949 mmol) and l-Ethyl-3-(3- Dimethylaminopropyl)carbodiimide (about 0.181 g, 0.949 mmol) were added at about 0 °C and stirred for about 10 minutes then methyl (S)-l-(2-((2S,3S)-3-amino- 2-hydroxy-4-phenylbutyl)-2-(biphenyl-4-ylmethyl)hydrazinyl)-3,3-dimethyl-l- oxosbutan-2-ylcarbamate (Intermediate 10, about 0.336 g, 0.632 mmol) in DCM (5 ml), DMF (2 ml) and NMM (0.35 ml, 3.164 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 24 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHCCH solution, brine and dried over Na2SC> , the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100-200 mesh, elution 50% EtOAC in hexane) to afford the title compound as an off white solid. Wt: 0.120 g; Yield: 24%; Ή NMR
(300 MHz, CDCb): δ 7.57-7.12 (m.20H), 6.54 (brs, lH), 5.71 (d, 1H, J = 8.1 Hz), 5.22 (d, 1H, J = 8.7 Hz), 5.15-4.99 (m, 4H), 4.86 (brs, 1H), 4.22-4.04 (m, 2H), 3.88- 3.74 (m, 2H), 3.62 (s, 3H), 3.64-3.55 (m, 1H), 2.91 (d, 1H, J = 7.2 Hz), 2.79-2.51 (m, 2H), 2.31-1.99 (m, 2H), 1.12 (s, 3H), 0.72 (s, 9H), 0.43 (s, 3H); Mass: [M+l]+ 792 (20%), [M+Na]+ 814 (100%); HPLC: 82.6%; IR (KBr, cm"1): 3419, 3030, 2957, 1709, 1513, 1369, 1346, 1255, 1158, 1108, 1055, 916, 826, 762, 699; M.R: 117.2 °C -120.8 °C.
Example 21: Preparation of methyl (S)-l-(2-((2S,3S)-2-hvdroxy-3-((lS.3R)-3- (methoxycarbonylarruno)-2,2-dimethylcyclobutanecarboxarnido)-4-phenylbutyl)-2- (4-(pyridin-2-yl)benzyl)hvdrazinyl)-4-methyl-l-oxopentan-2-ylcarbamate:
Figure imgf000071_0001
To a stirred solution (lS,3R)-3-(methoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 2, about 0.152 g, 0.758 mmol) in DCM (10 ml) at 0 °C hydroxybenzotriazole (0.145 g, 0.947 mmol) and l-Ethyl-3-(3-
Dimethylaminopropy carbodiimide (0.181 g, 0.947 mmol) were added and stirred about 10 minutes then methyl (S)-l-(2-((2S,3S)-3-amino-2-hydroxy-4-phenylbutyl)- 2-(4-(pyridin-2-yl)benzyl)hydrazinyl)-4-methyl-l-oxopentan-2-ylcarbamate
(Intermediate 11, about 0.336 g, 0.631 mmol) in DCM (10 ml) DMF (1 ml) and NMM (0.34 ml, 3.160 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 24 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHC03 solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100- 200 mesh, elution 50% EtOAC in hexane) to afford the title compound as a off white solid, which was recrystalized from DCM-Hexane and MeOH-water. Wt: 0.120 g; Yield: 26.5%; Ή NMR (300 MHz, CDC13): δ 8.68 (d, 1H, J = 4.5 Hz), 7.94 (d, 2H, J = 7.8 Hz), 7.81-7.68 (m, 2H), 7.40 (d, 1H, J = 8.1 Hz), 7.31-7.12 (m, 5H), 6.97 (s, 1H), 5.87 (d, 1H, J = 9 Hz), 5.10 (d, 1H, J = 8.7 Hz), 4.91-4.78 (m, 2H), 4.09-3.72 (m, 4H), 3.64, 3.61 (s, 6H), 3.57-3.49 (m, 1H), 2.92 (d, 2H, J = 7.5 Hz),
2.83-2.71 (m, 1H), 2.51-2.42 (m, 1H), 2.36-2.17 (m, 2H), 2.00-1.88 (m, lH), 1.40- 1.20 (m, 2H), 1.16 (s, 3H), 0.89-0.80 (m, lH), 0.72 (brs, 9 H); Mass: [M+l]+ 717 (15%), [M+Na]+739 (100%); HPLC: 98.26%; IR (KBr, cm'1): 3411, 3344, 2956, 1705, 1648, 1512, 1467, 1354, 1271, 1241, 1108, 1057, 781, 749; M.R: 208.6 °C - 210.9 °C. Example 22; Preparation of methyl (S)-l-(2-((2S,3S)-3-((lS.3R)-3-(tert- butoxycarbonylamjno)-2,2-dimethylcyclobutanecarboxamido)-2-hydroxy-4- phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hydrazinyD-4-methyl-l-oxopentan-2- ylcarbamate:
Figure imgf000072_0001
To a stirred solution (lS,3R)-3-(tert-butoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 3, about 0.156 g, 0.644 mmol) in DCM (10 ml) at 0 °C hydroxybenzotriazole (0.123 g, 0.805 mmol) and l-Ethyl-3-(3- Dimethylaminopropyl)carbodiimide (0.153 g, 0.805 mmol) were added and stirred for about 10 minutes then methyl (S)-l-(2-((2S,3S)-3-amino-2-hydroxy-4- phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hydrazinyl)-4-methyl-l-oxopentan-2- ylcarbamate (Intermediate 11, about 0.286 g, 0.537 mmol) in DCM (10 ml) DMF (1 ml) and NMM (0.29 ml, 2.685 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 24 hours. After completion of the reaction
(monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHCOj solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100- 200 mesh, elution 40% EtOAC in hexane) to afford the title compound as a white solid. Wt: 0.150 g; Yield: 36.76%; Ή NMR (300 MHz, CDCI3): δ 8.68 (d, 1H, J =
4.2 Hz), 7.94 (d, 2H, J = 8.1 Hz), 7.81-7.68 (m, 2H), 7.40 (d, 2H, J = 7.8 Hz), 7.29- 7.11 (m, 6H), 6.94 (s, 1H), 5.86 (d, 1H, J = 9.0 Hz), 4.92-4.74 (m, 3H), 4.10-3.98 (m, 1H), 3.94-3.81 (m, 2H), 3.80-3.69 (m, 1H), 3.61 (s, 3H), 3.58-3.48 (m, 1H), 2.92 (d, 1H, J = 7.5 Hz), 2.83-2.71 (m, 1H), 2.53-2.42(m, 1H), 2.35-2.13 (m, 2H), 1.99- 1.85 (m, 1H), 1.42 (s, 9H), 1.39-1.13 (m, 3H), 1.16 (s, 3H), 0.76 (brs, 9H); Mass:
[M+l]+ 759 (15%), [M+Na]+ 781 (100%); HPLC: 94.6% ;IR (KBr, cm1): 3354, 3060, 2957, 1706, 1702, 1698, 1506, 1510, 1513, 1467, 1453, 1437, 1385, 1366, 1254, 1170, 1051,989, 873, 779; M.R: 116.0 °C -125 °C. Example 23: Preparation of methyl (S)-l-(2-((2S,3S)-3-((lS.3R -3-acetamido-2,2- dimethylcvclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-(4-(pyridin-2- yl)benzyl)hydrazinyl)-4-methyl- 1 -oxopentan-2-ylcarbamate:
Figure imgf000073_0001
To a stirred solution (lS,3R)-3-acetamido-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 1, about 0.140 g, 0.758 mmol) in DMF (10 ml) at 0 °C hydroxybenzotriazole (0.145 g, 0.947 mmol) and l-Ethyl-3-(3- Dimethylaminopropyl)carbodiimide (0.181 g, 0.947 mmol) were added and stirred for about 10 minutes then methyl (S)-l-(2-((2S,3S)-3-amino-2-hydroxy-4- phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hydrazinyl)-4-methyl-l-oxopentan-2- ylcarbamate (Intermediate 11, about 0.336 g, 0.631 mmol) in DCM (10 ml) DMF (1 ml) and NMM (0.34 ml, 2.768 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 24 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHC03 solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100- 200 mesh, elution 90% EtOAC in hexane) and the obtained solid was recrystalized from DCM-hexane, MeOH-water to afford the title compound as an off white solid. Wt: 0.070 g; Yield: 15.8%; Ή NMR (300 MHz, CDCI3): δ 8.67 (d, 1H, J = 4.2 Hz), 7.93 (d, 2H, J = 8.1 Hz), 7.81-7.68 (m, 2H), 7.43-7.35 (d, 1H, J = 7.8 Hz), 7.32-7.12
(m, 6H), 6.94 (s, 1H), 6.06 (d, 1H, J = 8.4 Hz), 5.94 (d, 1H, J = 9.0 Hz), 4.86 (s, 1H), 4.79 (d, 1H, J = 7.5 Hz), 4.09-3.81 (m, 4H), 3.61 (s, 3H), 3.52-3.48 (m, 1H), 2.91 (d, 1H, J = 7.8 Hz), 2.84-2.70 (m, 1H), 2.52-2.43 (m, 1H), 2.37-2.24 (m, 2H), 1.97 (s, 3H), 1.97-1.88 (m, 1H), 1.38-1.19 (m, 3H), 1.18 (s, 3H), 0.99 (s, 3H), 0.74 (brs, 6H); Mass: [M+l]+701 (60%), [M+Na]+723 (100%); HPLC: 96.2%; IR (KBr, cm'1): 3419, 3323, 3061, 2957. 1710, 1702, 1656, 1649, 1529, 1467, 1437, 1369, 1271, 1156, 1113, 1055,828,780, 703; M.R: 115.0 °C -117.8 °C. Example 24: Preparation of methyl (S)-l-(2-((2S,3S)-2-hvdroxy-3-((lR.3S)-3-
(methoxycarbonylamino)-2,2-dimethylcvclobutanecarboxamido)-4-phenylbutyl)-2-
(4-(pyridin-2-yl)benzyl)hydrazinyl)-4-methyl-l-oxopentan-2-ylcarbamate:
Figure imgf000074_0001
To a stirred solution of (lR,3S)-3-(methoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 5, about 0.129 g, 0.644 mmol) in DCM (10 ml) at 0 °C hydroxybenzotriazole (0.109 g, 0.712 mmol) and l-Ethyl-3-(3- Dimethylaminopropyl)carbodiimide (0.136 g.0.712 mmol) were added and stirred for about 10 minutes then methyl (S)-l-(2-((2S,3S)-3-amino-2-hydroxy-4- phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hydrazinyl)-4-methyl-l-oxopentan-2- ylcarbamate (Intermediate 11, about 0.286 g, 0.537 mmol) in DCM (10 ml) DMF (1 ml) and NMM (0.24 ml, 2.369 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 48 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHC03 solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100- 200 mesh, elution 45% EtOAC in hexane) to afford the title compound as an off white solid which was again purified by recrystalization using DCM-he ane, MeOH - water and Isopropanol-diisopropyl ether combinations. Wt: 0.120 g; Yield: 31.2%; Ή NMR (300 MHz, CDC13): δ 8.68 (d, 1H, J = 4.2 Hz), 7.95 (d, 2H, J = 8.1 Hz),
7.80-7.67 (m, 2H), 7.42 (d, 2H, J = 8.1 Hz), 7.32-7.12 (m, 6H), 6.84 (brs, 1H), 5.74 (d, 1H, J = 8.1 Hz), 5.00 (d, 1H), 4.90-4.75 (m, 2H), 4.20-4.01 (m, 2H), 3.92-3.71 (m.3H), 3.61 (s, 6H), 2.91 (d, 1H, J = 7.8 Hz), 2.81-2.69 (m, lH), 2.64-2.53 (m, 1H), 2.32-2.11 (m, 2H), 2.09-1.93 (m, 1H), 1.39-1.18 (m, 3H), 1.14 (s, 3H), 0.80- 0.62 (m, 6H), 0.50 (s, 3H); Mass: [M+l]+ 717 (15%), [M+Na]+ 739 (100%); HPLC:
96.58%; IR (KBr, cm'1): 3419, 3332, 3062, 2956, 1702, 1507, 1517, 1467, 1456, 1354, 1259, 1195, 1155, 1111, 1054,989, 779, 747; M.R: 108.0-118.0 °C.
Example 25: Preparation of methyl (S)-l-(2-((2S.3S)-3-((lR,3S)-3-(tert- butoxycarbonylamino)-2,2-dimethylcyclobutanecarboxamido)-2-hvdroxy-4- phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hvdrazinyl)-4-methyl-l-oxopentan-2- ylcarbamate:
Figure imgf000075_0001
To a stirred solution of (lR,3S)-3-(tert-butoxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 6, about 0.161 g, 0.663 mmol) in
DCM (10 ml) at 0 °C hydroxybenzotriazole (about 0.127 g, 0.829 mmol) and 1- Ethyl-3-(3-Dimethylaminopropyl)carbodiimide (about 0.158 g, 0.829 mmol) were added and stirred for about 10 minutes then methyl (S)-l-(2-((2S,3S)-3-amino-2- hydroxy-4-phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hydrazinyl)-4-methyl-l- oxopentan-2-ylcarbamate (Intermediate 11, about 0.294 g, 0.552 mmol) in DCM (10 ml), DMF (1 ml) and NMM (about 0.3 ml, 2.764 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 48 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHCCh solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100-200 mesh, elution 90% EtOAC in hexane) to afford the title compound as an off white solid. Wt: 0.080 g; Yield: 19%; Ή NMR (300 MHz, CDCI3): δ 8.68 (d, 1H, J = 4.2 Hz), 7.94 (d, 2H, J = 8.1 Hz), 7.80-7.67 (m, 2H), 7.42 (d, 2H, J = 8.1 Hz), 7.36-7.12 (m, 6H), 6.87 (s, 1H), 5.74 (d, 1H, J = 8.4 Hz), 4.92-4.76 (m, 3H), 4.21-4.07 (m, 2H), 3.91-3.80 (m, 2H), 3.79-3.68 (m,
1H), 3.61 (s, 3H), 2.92 (d, 2H, J = 7.5 Hz), 2.84-2.70 (m, 1H), 2.63-2.54 (m, 1H), 2.31-2.12 (m, 2H), 2.09-1.93 (m, 1H), 1.41 (s, 9H), 1.30-1.18 (m, 3H), 1.13 (s, 3H), 0.80-0.62 (Brs, 6H), 0.49 (s, 3H); Mass: [M+l]+ 759 (25%), [M+Na]+ 781 (100%); HPLC: 89.22%; IR ( Br, cm"1): 3419, 3062, 3029, 2957, 1702, 1691, 1589, 1513, 1467, 1456, 1366, 1253, 1170, 1109, 1050, 780, 750; M.R: 115.2 °C -121.2 °C.
Example 26: Preparation of methyl (S)-l-(2-((2S,3S)-3-((lR,3S)-3-
(benzyloxycarbonylamino)-2,2-dimethylcyclobutanecarboxamido)-2-hydroxy-4- phenylbutyl)-2-(4-(pyridin-2-yDbenzyr)hvdrazinyr)-4-methyl-l-oxopentan-2- ylcarbamate:
Figure imgf000076_0001
To a stirred solution of (lR,3S)-3-(benzyloxycarbonylamino)-2,2- dimethylcyclobutanecarboxylic acid (Intermediate 4, about 0.194 g, 0.875 mmol) in DCM (20 ml) at 0 °C hydroxybenzotriazole (about 0.134 g, 0.700 mmol) and 1- Ethyl-3-(3-Dimethylaminopropyl)carbodiimide (about 0.167 g, 0.875 mmol) were added and stirred for about 10 minutes then methyl (S)-l-(2-((2S,3S)-3-amino-2- hydroxy-4-phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hydrazinyl)-4-methyl-l- oxopentan-2-ylcarbamate (Intermediate 11, about 0.311 g, 0.583 mmol) in DCM (20 ml), DMF (1 ml) and NMM (about 0.321 ml, 2.918 mmol) were added drop wise and the reaction mass was stirred at room temperature for about 24 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and washed with water, saturated NaHCO^ solution and brine and the organic layer was concentrated under reduced pressure, the resulting crude was purified by silica gel column (100-200 mesh, elution 90% EtOAC in hexane) to afford the title compound as an off white solid. Wt: 0.070 g; Yield: 15.1%; Ή NMR
(300 MHz, CDCI3): δ 8.66 (d, 1H), 7.98-7.92 (m, 2H), 7.79-7.67 (m, 2H), 7.46-7.13 (m, 13H), 6.99 (s, 1H), 5.77 (d, 1H, J = 9.0 Hz), 5.20-4.98 (m, 3H), 4.97-4.87 (m, 2H), 4.19-4.01 (m, 2H), 3.90-3.72 (m, 3H), 3.59 (s.3H).2.91 (d, 2H, J = 9.0 Hz), 2.80-2.69 (m, 1H), 2.62-2.54 (m, 1H), 2.30-2.14 (m, 2H), 2.10-1.99 (m, 1H), 1.29- 1.19 (m, 3H), 1.14 (s, 3H), 0.69 (brs, 6H), 0.48 (s, 3H); Mass: [M+l]+ 793 (15%),
[M+Na]+815 (100%); HPLC: 89.5%; IR (KBr, cm"1): 3426, 2928, 2956, 1692, 1524, 1467, 1341, 1259, 1156, 1121, 1052, 829, 779, 748; M.R: 106.5 °C -112.5 °C.
Example 27: Preparation of methyl N-[(lS)-l-(N'-[(2S,3S)-2-hydroxy-3-i(2S)-2- f(methoxycarbonyl)aminol-3,3-dimethylbutanamidol-4-phenylbutyn-N'-(i 4-Γ2-
(morpholin-4-yl)ethoxylphenyl ImethvQhydrazinecarbonyl 1-2,2- dimethylpiOpyllcarbamate:
Figure imgf000077_0001
Step 1: Synthesis of methyl N-[(lS)-l-(N'-[(2Sl3S)-3-([(tert- butoxy)carbonyl)amino}-2-hydroxy-4-phenylbutyl]-N'-{{4-[2-(morpholin-4- yl )ethoxy) phenyl Jmethy I )h hy I propyl) carbamate:
Figure imgf000077_0002
IPA (30 ml) was added to a mixture of (S)-l-((R)-oxiran-2-yl)-2- phenylethylcarbamate (0.626 g) and methyl N-[(lS)-2,2-dirnethyl-l-[N'-({4-[2- (moipholin-4-yl)ethoxy]phenyl }methyl)hydrazinecarbonyl]propyl]carbamate (Intermediate 15, 1.0 g) and the reaction mixture was stirred for 12 hours at refluxed temperature. After completion of the reaction (monitored by TLC), cooled the reaction mixture to room temperature, poured into an ice cold water, and collected the solid by filtration and dried the compound to get pure 800 mg of product (50 % Yield). Ή NMR (300 MHZ, CDC1?): δ 7.14-7.31 (7H. m), 6.84 (2H, d, J = 8.4 Hz), 6.52 (1H, brs), 5.30 (1H, d, J = 9.0 Hz), 5.08 (1H, d, J = 9.0 Hz), 4.70 (1H, brs), (4.06-4.10 (2H, m), 3.65-4.00 (5H, m), 3.64 (3H, s), 3.4-3.6 (2H, m), 2.77-2.81 (5H, m), 2.56-2.59 (4H, m), 1.36 (9H, s), 0.78 (9H, s); Mass: [M++H] 685 (100%).
Step 2: Synthesis of methyl N-[(lS)-l-{N'-[(2S,3S)-2-hydroxy-3-l(2S)-2- [(methoxycarbonyl)ainino]-3,3-dimethylbi anamido]-4-phenylbut\'l]-N'-({4-[2- (morpholin-4-yl)ethoxy] phenyl jmethyl)hydrazinecarbonylj -2,2- dimethylpropyl] carbamate :
Methyl N-[(lS)-l-{N'-[(2S,3S)-3-{ [(tert-butoxy)carbonyl]amino}-2- hydroxy-4-phenylbutyl]-N'-({4-[2-(morpholin-4- yl)ethoxy]phenyl}methyl)hydrazinecarbonyl }-2,2-dimethylpropyl]carbamate (Step 1, 400 mg) was taken in THF (8 ml) 4N HC1 (5 ml) was added and the reaction mixture was stirred at reflux temperature for 4 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure and azeotroped by using ethanol to get crude compound 384.3 mg which was used for next step without further purification.
(S)-methyl l-hydrazinyl-3,3-dimethyl-l-oxobutan-2-ylcarbamate (Step 3 for Intermediate 8, 132.43 g) was dissolved in dry DCM (20 ml) and 1 mL of DMF then it was cooled to 0°C and EDCI (167.44 mg), HOBT (134.13 mg) and NMM (294.8 mg) were added under nitrogen atmosphere. Then the above obtained crude free amino alcohol was added to the reaction mixture and the reaction mixture was stirred for over night at ambient temperature. After completion of the reaction (as monitored by TLC), the reaction mixture was diluted with DCM, washed with cold water and saturated NaHC03 solution followed by brine. The combined organic layers were dried over Na?S04, concentrated under reduced pressure to give crude compound which was further purified by column chromatography to afford 100 mg of pure compound along with a few amount of mixture. ER: In KBr(cm '): 3340, 2960, 1723, 1621, 1513, 1247, 1066; H1 NMR EN CDCL3: 0.819 (s, 9H), 0.889 (s, 9H), 2.38-2.43 (m, 1H), 2.44-2.48 (m, 4H), 2.58-2.60 (m, 3H), 2.80-2.91 (m, 2H),
3.51-3.95 (m, 12H), 4.08-4.12 (m,4 H), 5.22-5.35 (m, 2H), 6.451 (m, 2H), 6.82-6.85 (d, 2H, J=8.4Hz) 7.17-7.3 (m, 7H); MASS: 757[M+], [M+ Na+] 780.
Similarly, the following compounds have been prepared by the above corresponding procedures using their corresponding intermediates:
Example 28: Methyl N-f(lR)-l-(r(lS,2R)-l-hvdroxy-l-f(2S)-2-
[(methoxycarbonyl)aminol-3,3-dimethyl-N^(4-f2-(morpholin-4- yl)ethoxy1phenyl|methyl)butanehydrazido1-3-phenylpropan-2-yllcarbamoyl )-3- methylbutyllcarbamate:
Figure imgf000078_0001
ER: In Br(cm '): 3418, 2957, 1723, 1662. 1513, 1247, 1117, 1056; H1 NMR EN CDCL3: 0.722 (s, 9H), 0.861 (m, 6H), 1.4-1.9 (m, 3H), 1.92 (m, 3H), 2.082 (m, 2H), 2.16-2.788 (m, 3H), 2.788 (m, 4H), 2.849-2.945 (m, 5H), 3.56-4.32 (m, 16H), 6.83- 6.866 (d, 2H, J=8.4Hz), 7.15-7.24 (m, 7H); MASS: 771[M+], Example 29: Methyl N-f(lS)-l-fN'-i(lS,2R)-l-hvdroxy-2-(f(lS.3R)-3- [(methoxycarbonyl)arninol-2,2-dimethylcyclobutynformarnjdo)-3-phenylpropy N'-( ( 4-i2-(morpholin-4-yl)ethox ylphenyl I methvDhydrazinecarbon yl ) -2,2- dimethylpropyllcarbamate
Figure imgf000079_0001
IR: In KBr(cm '): 3333, 2957, 1710, 1675, 1513, 1456, 1368, 1249, 1116, 1056; H1 NMR IN CDCL3: 0.74-0.80 (m, 12H), 1.19(s, 3H), 1.97-2.07 (m, 6H), 2.17-2.28 (m, 3H,) 2.47- 2.53 (m, IH), 2.59-2.91 (m, 8H), 3.53-4.10 (m,12H),4.08- 4.12(m,4H)5.22-5.35 (m,2H), 6.451 (m, 2H), 6.82-6.85 (d, 2H. J=8.4Hz), 7.17-7.31 (m, 7H);MASS: 769[M+], [M+Na+]791.
Example 30: Methyl N-r(lS)-l-fN'-benzyl-N'-f(lS,2R)-2-if(lS,3R -3-(f(tert- butoxy)carbonyllamino ) -2,2-dimethylcyclobutyllformamido ) - 1 -hydroxy-3- phenylpropyllhvdrazinecarbonyl)-2,2-dimethylpropyllcarbamate:
Figure imgf000079_0002
ER: In KBr(cm'): 3426, 3356, 2962, 1709, 1691, 1514, 1509, 1367, 1253, 1171; H1 NMR IN CDCL3: 0.73-0.77 (m, 12H), 1.18 (s, 3H), 1.43 (s, 9H), 1.90-1.93 (m, IH), 2.21-2.28 (m, 2H), 2.50 (m, IH), 2.707-2.78 (m, IH), 2.89-2.91 (m, 6H), 3.57-3.60 (m, 2H), 3.67 (s, 3H), 3.67-3.99 (m, 4H), 4.80-4.83 (m, 2H), 5.24-5.27 (m, IH), 5.82 (s, IH), 6.51 (s, IH), 7.21-7.29 (m, 10H); MASS: 682 [M+], [M+ Na+] 705.
Example 31: Methyl N-r(lR.3S)-3-U(lS,2R)-l-hvdiOxy-l-{l-f(lR,3S)-3- [(methoxycarbonyl)aminol-2,2-dimethylcyclobutyn-N'-( [4-(pyridin-2- yPphenyllmethyl )formohydrazido)-3-phenylpropan-2-yllcarbamoyl 1-2,2- dimethylcyclobutyllcarbamat
Figure imgf000079_0003
IR: In KBr(cm"'): 3424, 3328, 2955, 1706, 1684, 1649, 1653, 1510, 1456, 1354, 1260, 1157, 1054, 1016, 779; H1 NMR IN CDCL3: 0.51 (s, 3H), 0.78 (s, 3H), 1.05 (s, 3H), 1.19 (s, 3H), 1.91-2.06 (m, 3H), 2.16-2.45 (m, 3H), 2.77 (d, IH, J=12Hz), 2.91 (m, 2H, J=7.5Hz), 3.31-3.49 (m, IH), 3.63 (s, 3H), 3.66 (s, 3H), 3.83-4.09 (m, 5H), 4.88 (brs, 2H), 4.98-5.16 (m, 2H), 5.89 (brs, 2H), 7.18-7.25 (m, 6H), 7.40 (d,
2H, J=8.1Hz), 7.62-7.78 (m, 2H), 7.94 (d, 2H, J=9.3Hz), 8.68 (d, IH, J=4.8Hz); MASS: 729[ΜΊ], [M+Na+] 751.
Example 32: Methyl N-f( 1 S )- 1 - ( N'-f( 1 S.2RV2- f K 1 S.3R>-3- ( idert- butoxy)carbon yllamino ) -2,2-dimethylcyclobutyllformamido ) - 1 -hydiOxy-3- phenylpropyn-N'-({4-f2-(morpholin-4- yDethoxylphenyl ) methyl )hydrazinecarbonyl )-2,2-dimethylpropyllcarbamate:
Figure imgf000080_0001
IR: In KBi(cm'): 3427, 3030, 2956, 2930, 2175, 1710, 1679, 1513. 1421, 1367, 1249, 1173; H1 NMR IN CDCL3: 0.76-0.80 (m, 12H), 1.19-1.43 (m, 3H), 1.91 (s, 9H), 2.27-2.44 (m, 5H), 2.56-2.59 (m, 5H), 2.78-2.82 (m, IH), 2.88-2.91 (m, 2H), 3.48-3.56 (m, 5H), 3.67 (s.3H), 3.73-3.88 (m, 6H), 4.06-4.10 (m, IH), 4.78-4.82 (m, IH), 5.23-5.26 (d, IH, J=9H), 5.82-5.84 (d, IH, J=8.7H), 6.82-6.85 (m, 3H), 7.17- 7.23 (m, 7H); MASS: [M+ Na+] 833.
Example 33: Methyl N-I(lS)-l-(N'-f(lS.2R)-2-(f(lR.3S)-3-(i(tert- butox y)carbonyllamino ) -2,2-dimethylcvclobutyllformarnido ) - 1 -h ydroxy-3- phenylpropyll-N'-( ( 4-[2-(morpholin-4- ypethoxylphenyl ImethyPhydrazinecarbonyl 1-2.2-dimethylpropyllcarbamate:
Figure imgf000080_0002
IR: In KBr(cm '): 3426, 3358, 2959, 1698, 1709, 1691, 1683, 1679, 1513, 1367, 1248, 1173, 1116; H1 NMR IN CDCL3: 0.44 (s, 3H), 0.76 (s, 9H).1.12 (s.9H), 1.41 (s, 3H), 2.03-2.25 (m, 3H), 2.56-2.90 (m, 9H), 3.59-4.17 (m, 13H), 4.77-4.82 (m, 2H), 5.24-5.27 (d, IH), 5.72-5.75 (d, 2H, J=9), 6.82-6.85(d, 2H, J=8.5), 7.18-7.24 (m, 7H); MASS: 811 [M+], [M+Na] 833.
Example 34: Methyl N-r(lS)-l-(N'-r(lS,2R)-l-hvdroxy-2-(i(lS,3R)-3- f(methoxycarbonyl)amino1-2,2-dimethylcvclobutynformamido)-3-phenylpropyll- N'-( ( 4-f 2-(morpholin-4-yl)ethoxy1phenyl ) meth vDhvdrazinecarbonyl } -2,2- dimethylpropyllcarbamate:
Figure imgf000081_0001
IR: In KBr(cm '): 3427, 2958, 1710, 1652, 1513, 1369, 1248, 1116; H1 NMR IN CDCL3: 0.743-0.906 (m, 15H), 1.21-1.26 (m, 3H), 1.98 (s, 3H), 2.29-2.32 (m, 2H),
2.47 (m, IH), 2.59-2.61 (m, 4H), 2.71 (m, 2H), 2.79-2.91 (m, 2H), 3.49-4.10 (m, 13H), 5.94 (d, IH), 6.089-6.11 (d, IH, J=9), 6.70 (s, IH), 6.81-6.84 (d, 2H, J=8.5), 7.15-7.25 (m, 6H); MASS: 753[M+), [M+Na] 775. Example 35: Preparation of methyl N-i(lR)-l-(i(lS.2R)-l-hydroxy-l-f(2S)-2- f (methoxycarbonyl )amino1-3,3-dimethyl-N'-( f 4-f 2-( 1 ,2,3,4-tetrahydroquinolin- 1 - yl)ethoxylphenyl)methyl)butanehydrazidol-3-phenylpropan-2-yllcarbamoyl 1-2,2- dimethylpropyllcarbamate:
Figure imgf000081_0002
Step 1: Synthesis of methyl N-{ Ί-[Ν'-(3-{ [(tert-butoxy )carbonyl] amino } -2-hydr
4-phenylbutyl)-N '-({4-[2-( 1,2,3, 4-tetrahydroquinolin- 1 '- yljethox}'] phenyl jmethyl)hydrazine 2-dimethylpropyl} carbamate
Figure imgf000081_0003
IPA (30 ml) was added to a mixture of (R)-methyl l-(2-(4-(2-(3, 4- dihydroquinolin-l(2H)-yl) ethoxy) benzyl) hydrazinyl)-3, 3-dimethyl-l-oxobutan-2- ylcarbamate (Intermediate 13, 1.0 g) and (S)-l-((R)-oxiran-2-yl)-2- phenylethylcarbamate (0.56 g) then the reaction mixture was stirred at reflux temperature for 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to room temperature and poured in to ice cold water. The obtained solid was filtered and dried to get the tittle compound (800 mg). H NMR (300 MHZ, CDC13): 0.80 (s, 9H); 1 .09 (s, 9H); 1.10- 1 .21 (m, 3H); 1.94- 1.97
(m, 3H); 2.74-2.93 (m, 5H); 3.41 -3.57 (m, 4H); 3.64 (s, 6H); 3.69-4. 14 (m, 6H); 4.78 (s, 1 H); 5.20-5.33 (m, 2H); 6.42 (s, 1 H); 6.55-6.63 (m, 2H); 6.80-6.96 (m, 2H); 7.02-7. 19 (m, 9H); Mass: (M+ l ) 732 ( 100 %). Step 2: Synthesis of methyl N-[(lR)-l-([(lS,2R)-l -hydroxy-l -[(2S)-2-
[(methox 'carbonyl)amino]-3,3-dimethyl-N'-({4-[2-( 1 ,2 ,3 ' ,4-tetrahydroquinolin- 1 - yl)ethoxy) 'phenyl }methyl)butanehydrazido] '- ' -phenylpropan-2-yl] 'carbamoyl) -2,2- di etl ylp ropy I ] ca rbamate :
To a solution of methyl N- { l -[N'-(3- { [(tert-butoxy)carbonyl]amino } -2- hydroxy-4-phenylbutyl)-N'-( ( 4-[2-( l ,2,3.4-tetrahydroquinolin- l - yl)ethoxy]phenyl ) methyl)hydrazinecarbonyl]-2,2-dimethylpropyl J carbamate (Step 1 , 400 mg) was taken in THF (8 ml) 4N HC1 (5 ml) was added. The reaction mixture was stirred at reflux temperature for 4 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure and azeotroped by using ethanol to get the crude compound then preceded for next step without further purification and characterization.
(S)-2-(methoxycarbonylamino)-3,3-dimethylbutanoic acid (Step- 1 for Intermediate 8, 132.4 mg) was dissolved in dry DCM (20 ml) and DMF ( 1 ml) then it was cooled to 0°C and EDCI ( 167.44 mg), HOBT ( 134. 13 mg), NMM (294.8 mg) was added under nitrogen atmosphere, then the above prepared compound was added to this mixture and the reaction mixture was stirred at room temperature for 18 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with ethyl acetate, washed with water, saturated sodium bicarbonate solution and brine solution, dried over Na2S04, and concentrated under reduced pressure and purified by column chromatography using hexane and ethyl acetate mixture as an eluent to afford the final compound. Ή NMR (300 MHz, CDC13): 0.80 (s, 9H); 0.88 (s, 9H); 1 . 10- 1 .21 (m, 3H); 1 .94- 1 .97 (m. 3H ); 2.74-2.93 (m, 5H); 3.41 -3.57 (m, 4H); 3.64 (s, 6H); 3.69-4. 14 (m, 6H); 4.78 (s, 1 H); 5.20-5.33 (m, 2H); 6.42 (s, 1H); 6.55-6.63 (m, 2H); 6.80-6.96 (m, 2H); 7.02-7.19 (m, 9H); ES Mass: 802 (100%), [M+l] 784; HPLC: 96.5 %.
Example 36: Preparation of methyl N-f(lR)-l-i iaS,2R)-l-i(2S)-N'-((4-i2-(4- ethylpiperazin-l-yl)ethoxylphenyl )methyl)-2-f(methoxycarbonyl)arrunol-3,3- dimethylbutanehydrazidol-l-hvdroxy-3-phenylpropan-2-yllcarbamoyl)-2,2- dimethylpropyllcarbamate:
Figure imgf000083_0001
Step 1: Synthesis of methyl N-{ 1 -[N'-(3-{[(tert-b toxy)carbonyl]cunino}-2-hydro 4-phenylb tyl)-N'-({4-[2-(4-ethylpiperazin-l- yl)ethoxy] phenyl }methyl)hydrazinecarbonyl) -2, 2-dimethylpropylj 'carbamate:
Figure imgf000083_0002
IPA (30 ml) was added to a mixture of (R)-methyl l-(2-(4-(2-(4- ethylpiperazin-l-yl) ethoxy) benzyl) hydrazinyl)-3, 3-dimethyl-l-oxobutan-2- ylcarbamate (Intermediate 12, 1.0 g) and tert-butyl (S)-l-((R)-oxiran-2-yl)-2- phenylethylcarbamate (0.58 g). The reaction mixture was stirred at reflux temperature for 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was cool to room temperature and poured in to ice cold water. The obtained solid was filtered and dried to get the tittle compound (800 mg). H NMR (300 MHZ, CDC13): 0.78 (s, 9H), 1.16-1.31 (m, 3H), 1.34 (s, 9H), 1.99-2.03 (m,
1H), 2.53-2.89 (m, 14H), 3.45-3.49 (m, 2H), 3.58 (s, 3H), 3.63-4.07 (m, 4H), 5.06- 5.09 (m, 1H), 5.28-5.36 (m, 1H), 6.78-6.81 (m. 3H), 7.19-7.21 (m. 7H); Mass: (M+Na) 735 (100 %).
Step 2; Synthesis of methyl N-[( 1 R)-l -/ [( 1 S,2R)-J -((2S)-N'-((4-[2-(4-ethylpiperazin- 1 -yljethoxy] phenyl jmethyl)-2f(methoxycarbonyl)amino] -3,3- dimethylbutanehydrazido] - 1 -hydroxy -3-pheny\propan-2-yl] 'carbamoyl} '-2,2- dimethylpropyl] carbamate
To a solution of methyl N-{l -[N'-(3-([(tert-butoxy)carbonyl]amino}-2- hydroxy-4-phenylb tyl)-N'-({4-[2-(4-ethylpiperazin-l- yl)ethoxy]phenyl}methyl)hydrazinecarbonyl]-2,2-dimethylpropyl}carbamate (Step 1 ,
400 mg) was taken in THF (8 ml) 4N HCI (5 ml) was added. The reaction mixture was stirred at reflux temperature for 4 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure and azeotroped by using ethanol to get the crude compound then preceded for next step without further purification and characterization.
(S)-2-(methoxycarbonylamino)-3,3-dimethylbutanoic acid (Step- 1 for Intermediate 8, 132.4 mg) was dissolved in dry DCM (20 ml) and DMF ( 1 ml) then it was cooled to 0°C and added EDCI ( 167.44 mg), HOBT ( 134. 13 mg), NMM (294.8 mg) under nitrogen atmosphere then the above prepared compound was added to this mixture and stirred the reaction at room temperature for 18 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with ethyl acetate, washed with water, saturated sodium bicarbonate solution and brine solution, dried over Na2S04, and concentrated under reduced pressure and purified by column chromatography using hexane and ethyl acetate mixture as an eluent to afford the final compound. Ή NMR (300 MHz, CDC13): 0.82 (s, 7H); 0.88
(s, 9H); 1.01 (s, 5H); 2.86-2.93 (m, 12H); 3.58-3.67 (m, 2H); 3.80 (m, 6H); 3.86- 4. 10 (m, 6H), 5.23-5.36 (m, 2H), 6.81 -6.84 (d, J=9Htz, 2H), 7. 18-7.20 (m, 7H). ES Mass: 783 ( 100%), [M+ l ] 784. IR cm': In KBr, 3427, 2959, 1722, 1513, 1510, 1247, 1063,
Similarly, the following compound has been prepared by above procedure using the corresponding Intermediate 14:
Example 37: Methyl N-f( l R )- l - ( f( l S.2R )- l -hvdroxy- l -f(2S)-2- [(methoxycarbonvDaminol-3,3-dimethyl-N'-( ( 4-[3-( 1 ,2,3,4-tetrahydroquinolin- 1 - yDpropoxylphenyl ) methyl )butanehydrazidol-3-phenylpropan-2-yllcarbamoyl ) -2,2- dimethylpropyllcarbamate:
Figure imgf000085_0001
Ή NMR (300 MHz, CDC13): 0.80(s, 9H); 0.88 (s, 9H); 1.10-1.21 (m, 3H); 1.94-1.97 (m, 3H); 2.74-2.93 (m, 5H); 3.41-3.57 (m, 6H); 3.64 (s, 6H); 3.69-4.14 (m, 6H); 4.78 (s, IH); 5.20-5.33 (m, 2H); 6.42 (s, IH); 6.55-6.63 (m, 2H); 6.80-6.96 (m, 2H); 7.02-7.19 (m,9H);ES Mass: 816 (100%), [M+l] 817.
Example 38: Methyl N-f( lS)-l-IN'-i(3S)-3-i(2S)-2-( r(tert-butoxy)carbonyllamino)- 4-methylpentanamidol-2-hydroxy-4-phenylbιltyll-N'-((4-[2-(mo holin-4- yl)ethoxy]phenyl )methyl)hydrazinecarbonyl)-2,2-dimethylpropyllcarbamate:
Figure imgf000085_0002
IR: In KBr(cm'): 3418, 2959, 1715, 1678, 1513, 1367, 1249, 1170, 1118, 1057; H1 NMR IN CDC13: 0.90 (s, 9H), 1-1.2 (m, 3H), 1.24-1.26 (m, 6H), 1.28 (s, 9H), 1.67- 1.69 (m, 5H), 2.45 (s, 2H), 2.47 (m, IH), 2.57-2.60 (m, 4H), 2.78-2.82 (m, 2H), 2.91-2.93 (m, 2H), 3.5 (s, 3H), 3.56-3.60 (m, 4H), 3.73-3.99 (m, 2H), 4.01-4.16 (m, 3H), 4.73-4.83 (m, IH), 5.24-5.27 (m, IH), 6.82-6.85 (d, 2H, J=9.3Hz), 7.18-7.25 (m, 7H); MASS: 799 [M+l], 821 [M+Na].
Pharmacological activity
The compounds described herein were tested for their anti-HIV activity following procedures given below. These protocols are illustrative and do not limit to the scope of the invention.
Example 39: Screening the activity of antivirals using Cell viability assay:
Reduction of 3-(4,5-dimethylthiozol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, sigma) is chosen as an optimal mend point of cell viability measurement (Mosmann, 1983; Cole, 1986; alley et al„ 1988). Cells (0.2xl06 cells per well) were seeded in 96-well plates. Increasing concentrations of compound were added to the cells and incubated at 37°C for about 14 hours in a C02 Incubator with 5% C02 The media was replaced with a fresh growth medium along with 20 μL· of 5 mg /ml 3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Sigma). After incubation for about 4 hours in a humidified atmosphere, the media was removed and 200 of 0. 1 N acidic isopropyl alcohol was added to the wells to dissolve the MTT-formazan crystals. The absorbance was recorded at 570 nm, immediately after the development of purple colour. Each experiment was conducted in triplicate and the data are represented as average, with standard deviation. Percent viability of the cells was computed with reference to the absorbance of reduced MTT in the experiments conducted in absence of any compound.
The above described examples were tested in the two biological assays as described above and some compounds were found following results as shown in table 1 .
Cell line Supt- 1 ; Virus: HIV- 1 -93IN 101 .
Table 1
Figure imgf000086_0001
Example 40: Screening the activity of antiviral :
Antiviral Assay:
MT2 cells were infected with HIV- 1 strain 92HT599 ( 10 TCID 50/30000 cells). The infected cells were plated at the concentration of -30000 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.
Compound Preparation:
l OmM stock was made by dissolving test compound in DMSO. Subsequent dilutions were made with DMSO to make necessary working stocks ( 100X) Cytotoxicity Assay:
For cytotoxicity assay the same amount of MT2 cells as in antivirus without HrV- 1 virus was added to the cytotoxicity plates. The cytotoxicity was measured using MTT reagent in parallel with P24 estimation. The percent viability is calculated in comparison with vehicle control.
The above described examples were tested in the two biological assays as described above and some compounds were found following results as shown in table 2 and 3.
Table 2
Figure imgf000087_0001
Table 3
Figure imgf000087_0002
References:
1 . Mosmami T, December 1983. Journal of immunological methods, 65 (1-2), 55-63.
2. SPC Cole, cancer chemotherapy and Pharmacology, 1986, 17, 259-263. 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

WE CLAIM :
1 . A compound of the formula
Figure imgf000089_0001
Formula (1 )
or a pharmaceutically acceptable salt, prodrug or stereoisomer there of, wherein,
Wi and 2 are independent and can be selected from a bond or substituted or unsubstituted alkylene and preferably the substi tuent is selected from n-butyl, isobutyl or terti ary butyl ;
Zi and Z2 are independent and can be selected from a bond or substituted or unsubstituted cyclo butyl ; provided that w hen atleast one of the Zi and Z2 is substituted or unsubstituted cyclo butyl then Ri can be H, -0[C(Ra)2]m-alkyl, -0[C(Rb)2]m- cycloalkyl, -0[C(Rc)2]m-heterocyclyl , substituted or unsubstituted phenyl, substituted or unsubstituted pyridi ne, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl ; or when both Zi and Z2 are a bond then Rj can be -0[C(Ra)2]n- Ria, wherei n Rla can be H, substituted or unsubstituted alky] , substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, or substi tuted or unsubstituted heteroaryl ;
R2, R3, R2a and R3a are i ndependent and can be selected from H, C(0)R", C(0)2R". or S(0)2R";
R4 can be H, OH, halogen, NR", C(0)2R", C(0)NR" substituted or unsubstituted alkyl , substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl ;
m can be an i nteger 0-3 ;
n can be an integer 1 -3;
Ra, Rb and Rc are independent and can be selected from H, or substi tuted or unsubstituted alkyl ;
R" can be H, substi tuted or unsubstituted alkyl , substituted or unsubstituted aryl , substi tuted or unsubstituted cycloalkyl , or substi tuted or unsubstituted heterocyclyl, an analog thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable solvate thereof, a pharmaceutically acceptable hydrate thereof, an N-oxide thereof, a tautomer thereof, a regioisomer thereof, a stereoisomer thereof, a prodrug thereof or a polymorph thereof.
2. A compound of the formula (IA):
Figure imgf000090_0001
Formula ( I A )
or a pharmaceutically acceptable salt, prodrug or stereoisomer there of, wherei n,
Wi and W2 are independent and can be selected from a bond or substituted or unsubstituted alkylene and preferably the substi tuent is selected from n-butyl, isobutyl or tertiary butyl ;
nx and ny are i ndependent and can be an i nteger 0- 1 and with the proviso that one of nx or ny is 1 ;
Ri can be H, -0[C(R3)2]m-alkyl , -0[C(Rb)2]m-cycloalkyl, -0[C(Rc)2]m- heterocyclyl, substituted or unsubstituted phenyl , substituted or unsubstituted pyridine, substituted or unsubstituted heterocyclyl , or substituted or unsubstituted heteroaryl ;
R2, R3, R2a and R3a are independent and can be selected from H, C(0)R", C(0)2R", or S(0)2R";
R4 can be H, OH, halogen, NR", C(0)2R", C(0)NR" substituted or unsubstituted alky] , substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl ;
R5, R6, R7, R8, Rsa, R a. ^7» and R8a are i ndependent and can be selected from H, substituted or unsubstituted alkyl , or substituted or unsubstituted cycloalkyl ;
m can be an integer 0-3 ;
Ra, Rb and Rc are i ndependent and can be selected from H, or substi tuted or unsubstituted alkyl ; R" can be H, substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted cycloalkyl, or substituted or unsubstituted heterocyclyl , an analog thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable solvate thereof, a pharmaceutically acceptable hydrate thereof, an N-oxide thereof, a tautomer thereof, a regioisomer thereof, a stereoisomer thereof, a prodrug thereof or a polymorph thereof.
3. A compound of the formula (IB):
Figure imgf000091_0001
Formula (IB )
or a pharmaceutically acceptable salt, prodrug or stereoisomer there of, wherein,
VVj and YV2 are independent and can be selected from substi tuted or unsubstituted alkylene and preferably the substituent is selected from n-butyl , isobutyl or tertiary butyl;
Ria can be H, substituted or unsubsti tuted alkyl , substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, or substi tuted or unsubstituted heteroaryl ;
R2, R3, R2a and R3a are independent and can be selected from H, C(0)R", C(0)2R", or S(0)2R";
R_, can be H, OH, halogen, NR", C(0)2R". C(0)NR" substituted or unsubstituted alkyl , substituted or unsubstituted alkyl, or substituted or unsubstituted cycloalkyl ;
n can be an integer 1 -3;
R" can be H, substi tuted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted cycloalkyl , or substituted or unsubsti tuted heterocycl yl , an analog thereof, a pharmaceutical ly acceptable salt thereof, a pharmaceutical ly acceptable solvate thereof, a pharmaceutically acceptable hydrate thereof, an N-oxide thereof, a tautomer thereof, a regioisomer thereof, a stereoisomer thereof, a prodrug thereof or a polymorph thereof.
4. A compound selected from the group consisting of:
( 1 S,3 )-3-acetamido-N-((2S,3S)-4-(2-(( 1 S,3R)-3-acetamido-2,2- dimethylcyclobutanecarbonyl)- 1 -(4-(pyridin-2-yl)benzyl)hydrazinyl)-3-hydroxy- 1 - phenylbutan-2-yl)-2,2-dimethylcyclobutanecarboxamide.
Methyl (lR)3S)-3-((2S,3S)-4-(2-((lS,3R)-3-methylcarbamato-2,2- dimethylcyclobutanecarbonyl)-l-(4-(pyridin-2-yl)benzyl)hydrazinyl)-3-hydroxy-l- phenylbutan-2-ylcarbamoyl)-2,2-dimethylcyclobutylcarbamate,
Tert-butyl (lS,3R)-3-((2S,3S)-4-(2-((lR,3S)-3-(tert-butyloxycarbonylamino)-2,2- dimethylcyclobutanecarbonyl)- l-(4-(pyridin-2-yl)benzyl)hydrazinyl)-3-hydroxy-l- phenylbutan-2-ylcarbamoyl)-2,2-dimethylcyclobutylcarbamate,
Benzyl (lS,3R)-3-((2S,3S)-4-(2-((lR,3S)-3-(benzyloxycarbonylamino)-2,2- dimethylcyclobutanecarbonyl)-l-(4-(pyridin-2-yl)benzyl)hydrazinyl)-3-hydroxy-l- phenylbutan-2-ylcarbamoyl)-2,2-dimethyIcyclobutylcarbamate,
Methyl (lS,3R)-3-((2S,3S)-3-hydroxy-4-(2-((lR,3S)-3-(methoxycarbonylamino)-
2.2- dimethylcyclobutanecarbonyl)-l-(4-(pyridin-2-yl)benzyl)hydrazinyl)-l-phenylbutan- 2-ylcarbamoyl)-2,2-dimethylcyclobutylcarbamate,
Methyl ( 1 R,3S)-3-((2S,3S)-3-hydroxy-4-(2-((S)-2-(methoxycarbonylamino)-3,3- dimethylbutanoyl)-l-(4-(pyridin-2-yl)benzyl)hydrazinyl)-l-phenylbutan-2-ylcarbamoyl)- 2,2-dimethylcyclobutylcarbamate,
Methyl (S)- 1 -(2-((2S,3S)-3-(( 1 S,3R)-3-(tert-butoxycarbonylamino)-2,2- dimethylcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-(4-(pyridin-2- yl)benzyl)hydrazinyl)-3,3-dimethyl- 1 -oxobutan-2-ylcarbamate,
Methyl (S)-l-(2-((2S,3S)-3-((lS,3R)-3-acetamido-2,2- dimethylcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-(4-(pyridin-2- yl)benzyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate,
Methyl (S)- 1 -(2-((2S,3S)-2-hydroxy-3-(( 1 R,3S)-3-(methoxycarbonylamino)-2,2- dimethylcyclobutanecarboxamido)-4-phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hydrazinyl)-
3.3- dimethyl-l-oxobutan-2-ylcarbamate, Methyl (S)-l-(2-((2S,3S)-3-((lR,3S)-3-(tert-butoxycarbonylamino)-2,2- dimethylcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-(4-(pyridin-2- yl)benzyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate,
Methyl (S)- 1 -(2-((2S,3S)-3-(( 1 R,3S)-3-(benzyloxycarbonylamino)-2,2- dimethylcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-(4-(pyridin-2- yl)benzyl)hydraziny])-3,3-dimethy]-l-oxobutan-2-ylcarbamate,
Methyl (S)-l-(2-benzy1-2-((2S,3S)-2-hydroxy-3-((lS,3R)-3- (methoxycarbonylamino)-2,2-dimethyIcyclobutanecarboxamido)-4- phenylbutyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate,
Methyl (S)-l-(2-((2S,3S)-3-((lS,3R)-3-acetamido-2,2- dimethylcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-benzylhydrazinyl)-3,3- di methyl- 1 -oxobutan-2-ylcarbamate,
Methyl (S)-l-(2-benzyl-2-((2S,3S)-3-((lS,3R)-3-(tert-butoxycarbonylamino)-2,2- dimethylcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)hydrazinyl)-3,3-dimethyl-l- oxobutan-2-ylcarbamate,
Methyl (S)- 1 -(2-((2S,3S)-3-(( 1 S,3R)-3-acetamido-2,2- dimethylcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-(biphenyl-4- ylmethyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate,
Methyl (S)-l-(2-(biphenyl-4-ylmethyl)-2-((2S,3S)-2-hydroxy-3-((lS,3R)-3- (methoxycarbonylamino)-2,2-dimethylcyclobutanecarboxamido)-4- phenylbutyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate,
Methyl (S)-l-(2-(biphenyl-4-ylmethyl)-2-((2S,3S)-3-((lS,3R)-3-(tert- butoxycarbonylamino)-2,2-dimethylcyclobutanecarboxamido)-2-hydroxy-4- phenylbutyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate,
Methyl (S)-l-(2-(biphenyl-4-ylmethyl)-2-((2S,3S)-2-hydroxy-3-((lR,3S)-3-
(methoxycajbonylamino)-2,2-dimethylcyclobutanecarboxamido)-4- phenylbutyl)hydrazinyl)-3,3-dimethyl-l -oxobutan-2-ylcarbamate,
Methyl (S)-l-(2-(biphenyl-4-ylmethyl)-2-((2S.3S)-3-((lR,3S)-3-(tert- butoxycarbonylamino)-2,2-dimethylcyclobutanecarboxamido)-2-hydroxy-4- phenylbutyl)hydrazinyl)-3,3-dimethyl- 1 -oxobutan-2-ylcarbamate, Methyl (S)-l-(2-((2S,3S)-3-((lR,3S)-3-(benzyloxycarbonylamino)-2,2- dimethyIcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-(biphenyl-4- ylmethyl)hydrazinyl)-3,3-dimethyl-l-oxobutan-2-ylcarbamate,
Methyl (S)-l-(2-((2S,3S)-2-hydroxy-3-((lS,3R)-3-(methoxycarbonylamino)-2,2- dimethylcyclobutanecarboxamido)-4-phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hydrazinyl)- 4-methyl- 1 -oxopentan-2-ylcarbamate,
Methyl (S)-l-(2-((2S,3S)-3-((lS,3R)-3-(tert-butoxycarbonylamino)-2,2- dimethylcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-(4-(pyridin-2- yl)benzyl)hydrazinyl)-4-methyl- 1 -oxopentan-2-ylcarbamate,
Methyl (S)-l-(2-((2S,3S)-3-((lS,3R)-3-acetamido-2,2- dimethylcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-(4-(pyridin-2- yl)benzyl)hydrazinyl)-4-methyl-l-oxopentan-2-ylcarbamate,
Methyl (S)-l-(2-((2S,3S)-2-hydroxy-3-((lR,3S)-3-(methoxycarbonylamino)-2,2- dimethylcyclobutanecarboxamido)-4-phenylbutyl)-2-(4-(pyridin-2-yl)benzyl)hydrazinyl)- 4-methyl- 1 -oxopentan-2-ylcarbamate,
Methyl (S)- 1 -(2-((2S,3S)-3-(( 1 R,3S)-3-(tert-butoxycarbonylamino)-2,2- dimethylcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-(4-(pyridin-2- yl)benzyl)hydrazinyl)-4-methyl-l-oxopentan-2-ylcarbamate,
Methyl (S)-l-(2-((2S,3S)-3-((lR,3S)-3-(benzy]oxycarbonylamino)-2,2- dimethylcyclobutanecarboxamido)-2-hydroxy-4-phenylbutyl)-2-(4-(pyridin-2- yl)benzyl)hydrazinyl)-4-methyl-l-oxopentan-2-ylcarbamate,
Methyl N-[(lS)-l-(N'-[(2S,3S)-2-hydroxy-3-[(2S)-2-[(methoxycarbonyl)amino]- 3,3-dimethylbutanamido]-4-phenylbutyI]-N'-({4-[2-(mo holin-4- yl)ethoxy]phenyl ) methyl)hydrazinecarbonyl } -2,2-dimethylpropyl]carbamate,
Methyl N-[( 1 R)- 1 - { [( 1 S,2R)- 1 -hydroxy- 1 -[(2S)-2-[(methoxycarbonyl)amino]-
3,3-dimethyl-N'-({4-[2-(moφholin-4-yl)ethoxy]phenyl }methyl)butanehydrazido]-3- phenylpropan-2-yl]carbamoyl }-3-methylbutyl]carbamate,
Methyl N-[(lS)-l-{N'-[(lS,2R)-l-hydroxy-2-{[(lS,3R)-3- [(methoxycarbonyl)amino]-2,2-dimethylcyclobutyl]formamido}-3-phenylpropyl]-N'-({4- [2-(morpholin-4-yl)ethoxy]phenyl }methyl)hydrazinecarbonyl }-2,2- dimethylpropyl]carbamate, Methyl N-[( 1 S)- 1 - { N'-benzyl-N'-[( 1 S,2R)-2- { [( 1 S,3R)-3- { [(tert- butoxy)carbonyl]amino)-2,2-dimethylcyclobuty]]formamido}-l-hydroxy-3- phenylpropyl]hydrazinecarbonyl } -2,2-dimethylpropyl]carbamate,
Methyl N-[(lR,3S)-3-{ [(lS,2R)-l-hydroxy-l-{ 1 -[( 1 R,3S)-3- [(methoxycarbonyl)amino]-2,2-dimethylcyclobutyl]-N'-{ [4-(pyridin-2- yl)phenyl]methyl)formohydrazido)-3-phenylpropan-2-yl]carbamoyl}-2,2- dimethylcyclobutyl]carbamate,
Methyl N-[(lS)-l-{N'-[(lS,2R)-2-{ [(lS,3R)-3-{[(tert-butoxy)carbonyl]amino)- 2,2-dirnethylcyclobutyl]formamido}-l-hydroxy-3-phenylpropyl]-N'-({4-[2-(moφholin-4- yl)ethoxy]phenyl}methyl)hydrazinecarbonyl }-2,2-dimethy]propyl]carbamate,
Methyl N-[(lS)-l-{N'-[(lS,2R)-2-{[(lR,3S)-3-{[(tert-butoxy)carbonyl]amino}-
2.2- dimethy)cyclobutyl]formamido}-l-hydroxy-3-phenylpropyl]-N'-({4-[2-(mo holin-4- yl)ethoxy]phenyl } methyl )hydrazinecarbonyl }-2,2-dimethylpropyl]carbamate.
Methyl N-[(lS)-l-{N (lS,2R)-l-hydroxy-2-{[(lS,3R)-3- [(methoxycarbonyl)amino]-2,2-dimethylcyclobutyl]formamido}-3-phenyIpropyl]-N'-({4- [2-(moφholin-4-yl)ethoxy]phenyl } methyl )hydrazi necarbonyl ) -2,2- dimethylpropyl]carbamate,
Methyl N-[(lR)-l-{ [(IS, 2R)-1 -hydroxy- 1 -[(2S)-2-[(methoxycarbony l)amino]-
3.3- dimethyl-N'-({4-[2-(l,2,3,4-tetrahydroquinolin-l- yl)ethoxy]phenyl }methyl)butanehydrazido]-3-phenylpropan-2-yl]carbamoyl }-2,2- dimethylpropyl]carbamate,
Methyl N-[(l R)-l-{ [(1 S,2R)-l-[(2S)-N'-({4-[2-(4-ethylpiperazin-l- yl)ethoxy]phenyl }methyl)-2-[(methoxycarbonyl)amino]-3,3-dimethylbutanehydrazido]- l-hydroxy-3-phenylpropan-2-yl]carbamoyl }-2,2-dimethylpropyl]carbamate,
Methyl N-[(lR)-l-([(lS,2R)-1-hydroxy-l-[(2S)-2-[(methoxycarbonyl)amino]-
3,3-dimethyl-N'-({4-[3-(l,2,3,
4-tetrahydroquinolin-l- yl)propoxy]phenyl}methyl)butanehydrazido]-3-phenylpropan-2-yl]carbamoyl }-2,2- dimethylpropyl]carbamate.
Methyl N-[(lS)-l-(N,-[(3S)-3-[(2S)-2-{[(tert-butoxy)carbonyl]amino}-4- methylpentanamido]-2-hydroxy-4-phenylbutyl]-N,-({4-[2-(moφholin-4- yl)ethoxy]phenyl}methyl)hydrazinecarbonyl}-2,2-dimethylpropyl]carbamate, and pharmaceutically acceptable salts thereof.
5. A pharmaceutical composition comprising a compound according to any one of claims 1-4 and a pharmaceutically acceptable excipient.
6. The pharmaceutical composition according to claim 5, wherein the pharmaceutically acceptable excipient is a carrier or diluent.
7. 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 1-4.
8. The method according to claim 7, wherein the viral mediated disease, disorder or syndrome is selected from the group consisting of HIV infection and a retroviral infection genetically related to AIDS.
9. 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 1-4.
10. 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 claim 5.
11. The method according to claim 10, wherein the viral mediated disease, disorder or syndrome is selected from the group consisting of HIV infection and a retroviral infection genetically related to AIDS.
PCT/IB2010/003313 2009-12-29 2010-12-20 Novel aza-peptides containing 2,2-disubstituted cyclobutyl and/or substituted alkoxy benzyl derivatives as antivirals Ceased WO2011080562A1 (en)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10294234B2 (en) 2017-02-06 2019-05-21 Gilead Sciences, Inc. HIV inhibitor compounds
US11052087B2 (en) 2018-07-30 2021-07-06 Gilead Sciences, Inc. Anti-HIV compounds
CN113603634A (en) * 2021-08-06 2021-11-05 江苏八巨药业有限公司 Preparation method of atazanavir intermediate
CN114890916A (en) * 2022-04-25 2022-08-12 常州吉恩药业有限公司 A kind of preparation method of N-methoxycarbonyl-L-tertiary leucine

Citations (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4598095A (en) 1982-03-05 1986-07-01 Tamio Nishimura Antiviral isothiosemicarbazones
WO1994019332A1 (en) 1993-02-25 1994-09-01 Abbott Laboratories Retroviral protease inhibiting compounds
EP0724650A1 (en) 1993-07-09 1996-08-07 University Of Pittsburgh Of The Commonwealth System Of Higher Education Selection and use of antiviral peptides
WO1997019055A1 (en) 1995-11-21 1997-05-29 Novartis Ag Azahexane derivatives as substrate isosters of retroviral asparate proteases
WO1997040029A1 (en) 1996-04-22 1997-10-30 Novartis Ag Antivirally active heterocyclic azahexane derivatives
WO1998003476A1 (en) 1996-07-17 1998-01-29 Novartis Ag Anilinopeptide derivatives
US5753652A (en) * 1991-07-03 1998-05-19 Novartis Corporation Antiretroviral hydrazine derivatives
US5849911A (en) 1996-04-22 1998-12-15 Novartis Finance Corporation Antivirally active heterocyclic azahexane derivatives
EP0989862A1 (en) 1997-06-23 2000-04-05 N-Gene Research Laboratories Inc. Pharmaceutical composition with antiviral activity containing an hydroxymic acid derivative and an antiviral agent
US6087383A (en) 1998-01-20 2000-07-11 Bristol-Myers Squibb Company Bisulfate salt of HIV protease inhibitor
WO2001007646A2 (en) 1999-07-21 2001-02-01 Martin Heinkelein Method for quantization of the antiviral effect of antiviral active principles
WO2001065957A2 (en) 2000-03-07 2001-09-13 Albert David M Grip-enhancing glove
US20020068757A1 (en) 1995-06-23 2002-06-06 Advanced Life Sciences, Inc. Biflavanoids and derivatives thereof as antiviral agents
WO2003037908A1 (en) 2001-10-31 2003-05-08 Ribapharm Inc. Antiviral combination therapy and compositions
WO2008011117A2 (en) * 2006-07-21 2008-01-24 Gilead Sciences, Inc. Antiviral protease inhibitors
WO2008156632A1 (en) 2007-06-12 2008-12-24 Concert Pharmaceuticals, Inc. Azapeptide derivatives

Patent Citations (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4598095A (en) 1982-03-05 1986-07-01 Tamio Nishimura Antiviral isothiosemicarbazones
US5753652A (en) * 1991-07-03 1998-05-19 Novartis Corporation Antiretroviral hydrazine derivatives
WO1994019332A1 (en) 1993-02-25 1994-09-01 Abbott Laboratories Retroviral protease inhibiting compounds
EP0724650A1 (en) 1993-07-09 1996-08-07 University Of Pittsburgh Of The Commonwealth System Of Higher Education Selection and use of antiviral peptides
US20020068757A1 (en) 1995-06-23 2002-06-06 Advanced Life Sciences, Inc. Biflavanoids and derivatives thereof as antiviral agents
WO1997019055A1 (en) 1995-11-21 1997-05-29 Novartis Ag Azahexane derivatives as substrate isosters of retroviral asparate proteases
WO1997040029A1 (en) 1996-04-22 1997-10-30 Novartis Ag Antivirally active heterocyclic azahexane derivatives
US5849911A (en) 1996-04-22 1998-12-15 Novartis Finance Corporation Antivirally active heterocyclic azahexane derivatives
WO1998003476A1 (en) 1996-07-17 1998-01-29 Novartis Ag Anilinopeptide derivatives
EP0989862A1 (en) 1997-06-23 2000-04-05 N-Gene Research Laboratories Inc. Pharmaceutical composition with antiviral activity containing an hydroxymic acid derivative and an antiviral agent
US6087383A (en) 1998-01-20 2000-07-11 Bristol-Myers Squibb Company Bisulfate salt of HIV protease inhibitor
WO2001007646A2 (en) 1999-07-21 2001-02-01 Martin Heinkelein Method for quantization of the antiviral effect of antiviral active principles
WO2001065957A2 (en) 2000-03-07 2001-09-13 Albert David M Grip-enhancing glove
WO2003037908A1 (en) 2001-10-31 2003-05-08 Ribapharm Inc. Antiviral combination therapy and compositions
WO2008011117A2 (en) * 2006-07-21 2008-01-24 Gilead Sciences, Inc. Antiviral protease inhibitors
WO2008156632A1 (en) 2007-06-12 2008-12-24 Concert Pharmaceuticals, Inc. Azapeptide derivatives

Non-Patent Citations (26)

* Cited by examiner, † Cited by third party
Title
"Bioreversible Carriers in Drug Design", 1987, AMERICAN PHARMACEUTICAL ASSOCIATION AND PERGAMON PRESS
"Remington: The Science and Practice of Pharmacy", 2003, LIPPINCOTT WILLIAMS & WILKINS
AVERETT, D.R.: "Anti-HIV compound assessment by two novel high capacity assays", J.VIROL.METHODS, vol. 23, 1989, pages 263 - 276, XP023788447, DOI: doi:10.1016/0166-0934(89)90159-6
BALZARINI ET AL., AIDS, vol. 5, 1991, pages 21 - 28
BIO ORG. MED. CHEM. LETT., vol. 3, 1993, pages 2837 - 2842
BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, vol. 16, no. 6, 2006, pages 1712 - 1715
CONNOR ET AL., JOURNAL OF VIROLOGY, vol. 70, 1996, pages 5306 - 5311
DALUGE, S. M. ET AL.: "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 CHEMOTHERA, vol. 38, no. 7, 1994, pages 1590 - 1603
ERICE ET AL., ANTIMICROB. AG. CHEMOTHERAPY, vol. 37, 1993, pages 385 - 383
FEDYUK N.V. ET AL., PROBLEMS OF VIROLOGY, 1992, pages 135
H.MITSUYA; 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, vol. 83, 1986, pages 1911 - 15
HARRIGTON ET AL., JOURNAL OF VIROLOGY METHODS, vol. 88, 2000, pages 111 - 115
J. ORG. CHEM., vol. 52, 1987, pages 689
KOYANAGI ET AL., INT. J. CANCER, vol. 36, 1985, pages 445 - 451
MEEK T.D ET AL.: "Inhibition of HIV-1 protease in infected T-limphocytes by synthetic peptide analogues", NATURE, vol. 343, 1990, pages 90, XP000083795, DOI: doi:10.1038/343090a0
MOSMANN T, JOURNAL OF IMMUNOLOGICAL METHODS, vol. 65, no. 1-2, December 1983 (1983-12-01), pages 55 - 63
PENNINGTON ET AL., PEPTIDES, 1990
POPIK ET AL., JOURNAL OF VIROLOGY, vol. 76, 2002, pages 4709 - 4722
RODA RANI ET AL., ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, vol. 456, no. 1, 1 December 2006 (2006-12-01), pages 79 - 92
ROOS ET AL., VIROLOGY, vol. 273, 2000, pages 307 - 315
SCHWARTZ, O. ET AL.: "A rapid and simple colorimeric test fror the study of anti HIV agents", AIDS RES. AND HUMAN RETROVIRUSES, vol. 4, no. 6, 1998, pages 441 - 447
SPC COLE, CANCER CHEMOTHERAPY AND PHARMACOLOGY, vol. 17, 1986, pages 259 - 263
T. HIGUCHI; W. STELLA: "Pro-drugs as Novel Delivery Systems", A.C.S. SYMPOSIUM SERIES, vol. 14
T.MIMOTO ET AL., J.MED. CHEM., vol. 42, 1999, pages 1789 - 1802
UCKUN ET AL., ANTIMICOBIAL AGENTS AND CHEMOTHERAPY, vol. 42, 1998, pages 383
WEISLOW ET AL., J.NATL.CANCER INST., vol. 81, 1989, pages 577 - 586

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US11078208B1 (en) 2017-02-06 2021-08-03 Gilead Sciences, Inc. HIV inhibitor compounds
US12084455B2 (en) 2017-02-06 2024-09-10 Gilead Sciences, Inc. HIV inhibitor compounds
US12479853B2 (en) 2017-02-06 2025-11-25 Gilead Sciences, Inc. HIV inhibitor compounds
US11052087B2 (en) 2018-07-30 2021-07-06 Gilead Sciences, Inc. Anti-HIV compounds
CN113603634A (en) * 2021-08-06 2021-11-05 江苏八巨药业有限公司 Preparation method of atazanavir intermediate
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