Field of the Invention
The present invention relates to cell adhesion inhibitors The compounds of this invention can be useful for inhibition and prevention of cell adhesion and cell adhesion mediated pathologies including inflammatory and autoimmune diseases such as bronchial asthma, rheumatoid arthritis, type I diabetes, multiple sclerosis, allograft rejection or psoriasis This invention also relates to pharmacological compositions containing the compounds of the present invention, and methods of treating bronchial asthma, rheumatoid arthritis, multiple sclerosis, type I diabetes, psoriasis, allograft rejection, and other inflammatory and/or autoimmune disorders using the compounds
Background of the Invention
Cell adhesion is a process by which cells associate with each other, migrate towards a specific target or localize within the extra-cellular matrix These interactions are mediated by specialized molecules called cell adhesion molecules (CAMs) CAMs have been demonstrated to participate in various cell-cell, cell-extracellular matrix, and platelet- platelet interactions They influence the adhesion of leukocytes to the vascular endothelium, their transendothehal migration, retention at extravascular sites and activation of T cells and eosinophils These processes are central to the pathogenesis of inflammatory and autoimmune diseases Therefore, CAMs are considered as potential targets to treat such disorders
CAMs can be classified into three groups - integrins, selectins and the immunoglobulin superfamily Out of these, integrins are key mediators in the adhesive interactions between hemopoietic cells and their microenvironment They are comprised of alpha-beta heterodimers that integrate signals from outside to the inside of cells and vice versa Integrins can be classified on the basis of the beta subumts they contain For example, beta-1 subfamily contains beta-1 subunit noncovalently linked to one of the 10 different alpha subumts
The alpha-4 beta-1 integrin, also known as VLA-4 (very late activation antigen 4), is a member of beta 1 integrin family and comprises of alpha-4 and beta-1 subumts It interacts with two specific ligands - the vascular cell adhesion molecule (VCAM-1) and the CS1 region of the protein fibronectin Adhesion mediated by VLA-4 is central to the process of
transendothehal migration of leukocytes Ligation of VLA-4 is followed by gross rearrangement of the cytoskeleton leading to flattening of cells along the blood vessel wall followed by expression of specific molecules, which digest the endothelial cell wall and diapedesis Once in the extraluminal region, the interactions of VLA-4 with extracellular fibronectin play a crucial role in migration to the site of inflammation, T cell proliferation, expression of cytokines and inflammatory mediators In addition, VLA-4 ligation provides co-stimulatory signals to the leukocytes, resulting in enhanced immunoreactivity Therefore, it is expected that VLA-4 antagonists would ameliorate the immune response through twofold actions - inhibition of T cell recruitment at the site of inflammation and inhibition of costimulatory activation of immune cells
Inhibitors of VLA-4 interactions have demonstrated beneficial therapeutic effects in several animal models of inflammatory, and allergic diseases including sheep allergic asthma (Abraham et al, J Clin Invest, 93, 776 (1994)), arthritis (Wahl et al, J Clin Invest 94, 655 (1994)), experimental allergic encephomyehtis (Yednock et al, Nature (Lond), 356, 63 (1992) and Baron et al, J Exp Med , 177, 57 (1993)), contact hypersensitivity (Chisolm et al Eur J Immunol, 23,682 (1993)), type I diabetes (Yang etaLProc Natl Acad Sci (USA), 90, 10494 (1993)) and inflammatory bowel disease (Podolsky et al, J Clin Invest, 92, 372(1993))
Region of CS1 moiety of fibronectin involved in the interaction with VLA-4 was identified as the tnpeptide Leu-Asp-Val, also known as LDV (Komonya et al, J Biol Chem 266, 15075(1991)) Taking a lead from this, several peptides containing the LDV sequence were synthesized which have shown to inhibit the in vivo interaction of VLA-4 to its hgands (Ferguson et al, Proc Natl Acad Sci (USA), 88, 8072 (1991), Wahl et al, J Clin Invest, 94, 655(1994), Nowhn et al, J Biol Chem , 268(27), 20352(1993) and PCT application PCT/US 91/04862
Despite these advances, there remains a need for inhibitors of VLA-4 dependent cell adhesion molecules New generations of molecules with oral efficacy would provide useful agents for treatment, prevention or suppression of various inflammatory pathologies mediated by VLA-4 binding
An article in Ann Rep Med Chem , 37, (2002) p 65, summarizes the highlights of work in the area of VLA-4 biology and small molecule antagonists
WO 01/12186 discloses non-peptidic compounds, which have been reported to inhibit the binding of hgands to VLA-4 WO 00/71572 and WO 99/26921 disclose (3-alanine derivatives which have been said to be the antagonists of VLA-4 and/or a4p7 WO 99/54321 discloses substituted diamines, which have been reported to have the ability to regulate the interaction of VCAM-1 and fibronectin with the mtegrin VLA-4 (a4/pi) WO 98/53817 discloses biarylalkanoic acids, which have been reported to be useful as cell adhesion inhibitors US 5,340,801 discloses N-arylcarbamoyl proline analogues, which have been said to be useful as cholecytokinin and gastrin antagonists WO 98/53814 discloses heterocyclic amide compounds, which have been described as cell adhesion inhibitors WO 99/67230 discloses compounds, which have been said to be inhibitors of oc4/pi mediated adhesion to either VCAM or CS-1
Summary of the Invention
The present invention provides cell adhesion inhibitors Pharmaceutically acceptable salts, pharmaceutically acceptable solvates, stereoisomers, tautomers, racemates, regioisomers, prodrugs, metabolites, polymorphs or N-oxides of these compounds having the same type of activity are also provided
Compounds provided herein were screened for inhibitory activity in a VLA-4 mediated cell adhesion assay These compounds could be used in treatment of chronic, cell adhesion mediated, allergic, autoimmune and inflammatory disorders, such as bronchial asthma, multiple sclerosis, rheumatoid arthritis etc
Pharmaceutical composition containing the compounds, and which may also contain pharmaceutically acceptable carriers or diluents, can be used for the treatment of cell adhesion mediated pathologies, including inflammatory and autoimmune diseases such as bronchial asthma, rheumatoid arthritis, type I diabetes, multiple sclerosis, allograft rejection or psoriasis
Other objects will be set forth in accompanying description which follows and in the part will be apparent from the description or may be learnt by the practice of the invention
In accordance with one aspect of the invention there are provided compounds having the structure of Formula I
and its pharmaceutically acceptable salts, pharmaceutically acceptable solvates, stereoisomers, tautomers, racemates, regioisomers, prodrugs, metabolites, polymorphs or N-oxides wherein ring A including X can be a five membered ring wherein X can be nitrogen, Y can be -CONH-,
Ri can be aryl (optionally substituted with alkyl, aryl or alkoxy) or aralkyl (wherein aryl group can be fused with a cycloalkyl or heterocyclyl group),
R2 can be nitro, amino, hydroxy, halogen, -NHCOR3 (wherein R3 can be optionally substituted heteroaryl, aryl, alkyl or heterocyclyl, -NHSO2R4 (wherein R4 can be optionally substituted alkyl or aryl) or -O (CH2) 2 R5 (wherein R5 can be (un) substituted heterocyclyl)
The following definitions apply to terms as used herein
The term "alkyl," unless otherwise specified, refers to a monoradical branched or
unbranched saturated hydrocarbon chain having from 1 to 20 carbon atoms This term can be
exemplified by groups such as methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-
butyl, t-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, n-decyl, tetradecyl, and the like Alkyl
groups may be substituted further with one or more substituents selected from alkenyl,
alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino, acyloxy, alkoxycarbonylamino,
azido, cyano, halogen, hydroxy, oxo, thiocarbonyl, carboxy, carboxyalkyl, aryl, heterocyclyl,
heteroaryl, arylthio, thiol, alkylthio, aryloxy, nitro, aminosulfonyl, aminocarbonylamino, -
NHC(=O)Rf, -NRfRq, -C(=O)NR„ -NHC(=O)NRfRq, -C(=O)heteroaryl,
C(=O)heterocyclyl, -0-C(=O)NRfRq {wherein Rf and Rq are independently selected from alkyl, alkenyl, cycloalkyl, cycloalkenyl, aryl, aralkyl, heterocyclyl, heteroaryl, heterocyclylalkyl, heteroarylalkyl}, nitro, or -SO2R60 (wherein R60 is alkyl, alkenyl, alkynyl, cycloalkyl, aralkyl, aryl, heterocyclyl, heteroaryl, heteroarylalkyl or heterocyclylalkyl) Unless otherwise constrained by the definition, alkyl substituents may be further substituted
by 1-3 substituents selected from alkyl, carboxy, -NRfRq, -C(=O)NRfRq, -OC(=O) NRfRq -NHC(=O)NRfRq (wherein Rf and Rq are the same as defined earlier), hydroxy, alkoxy, halogen, CF3, cyano, and -SO2R60, (wherein R60 are the same as defined earlier), or an alkyl group also may be interrupted by 1-5 atoms of groups independently selected from oxygen, sulfur or -NRa- {wherein Ra is selected from hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, acyl, aralkyl,-C(=O)ORf (wherein Rf is the same as defined earlier), SO2R60 (where R60 is as defined earlier), or -C(=O)NRfRq (wherein Rf and Rq are as defined earlier)} Unless otherwise constrained by the definition, all substituents may be substituted further by 1-3 substituents selected from alkyl, carboxy, -NRfRq, -C (=O)NRfRq, -0-C(=O)NRfRq (wherein Rf and Rq are the same as defined earlier) hydroxy, alkoxy, halogen, CF3, cyano, and -SO2R60 (where R60 is same as defined earlier), or an alkyl group as defined above that has both substituents as defined above and is also interrupted by 1-5 atoms or groups as defined above
The term "alkenyl," unless otherwise specified, refers to a monoradical of a branched or unbranched unsaturated hydrocarbon group having from 2 to 20 carbon atoms with cis, trans, or geminal geometry In the event that alkenyl is attached to a heteroatom, the double bond cannot be alpha to the heteroatom Alkenyl groups may be substituted further with one or more substituents selected from alkyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino, acyloxy, -NHC (=O)Rf, -NRfRq, -C(=O)NRfRq, -NHC(=O)NRfRq, -O-C(=O)NRfRq (wherein Rf and Rq are the same as defined earlier), alkoxycarbonylammo, azido, cyano, halogen, hydroxy, oxo, thiocarbonyl, carboxy, arylthio, thiol, alkylthio, aryl, aralkyl, aryloxy, heterocyclyl, heteroaryl, heterocyclyl alkyl, heteroaryl alkyl, ammosulfonyl, aminocarbonylamino, alkoxyamino, nitro, or SO2R60 (wherein R60 is same as defined earlier) Unless otherwise constrained by the definition, alkenyl substituents optionally may be substituted further by 1-3 substituents selected from alkyl, carboxy, hydroxy, alkoxy, halogen, -CF3, cyano, -NRfRq, -C(=O)NRfRq, -0-C(=O)NRfRq (wherein Rf and Rq are the same as defined earlier) and -SO2R60 (where R60 is same as defined earlier)
The term "alkynyl," unless otherwise specified, refers to a monoradical of an unsaturated hydrocarbon, having from 2 to 20 carbon atoms In the event that alkynyl is attached to a heteroatom, the triple bond cannot be alpha to the heteroatom Alkynyl groups may be substituted further with one or more substituents selected from alkyl, alkenyl, alkoxy,
cycloalkyl, cycloalkenyl, acyl, acylamino, acyloxy, alkoxycarbonylamino, azido, cyano, halogen, hydroxy, oxo, thiocarbonyl, carboxy, arylthio, thiol, alkylthio, aryl, aralkyl, aryloxy, aminosulfonyl, aminocarbonylamino, nitro, heterocyclyl, heteroaryl, heterocyclylalkyl, heteroarylalkyl, -NHC(=O)Rf -NRfRq, -NHC(=O)NRfRq , -C(=O)NRfRq, -0-C(=O)NRfRq (wherein Rf and Rq are the same as defined earlier), or -SO2R60 (wherein R60 is as defined earlier) Unless otherwise constrained by the definition, alkynyl substituents optionally may be substituted further by 1-3 substituents selected from alkyl, carboxy, carboxyalkyl, hydroxy, alkoxy, halogen, CF3, -NRfRq, -C(=O)NRfRq, -NHC(=O)NRfRq , -C(=O)NRfRq (wherein Rf and Rq are the same as defined earlier), cyano, or -SO2R60 (where R60 is same as defined earlier)
The term "cycloalkyl," unless otherwise specified, refers to cyclic alkyl groups of
from 3 to 20 carbon atoms having a single cyclic ring or multiple condensed rings, which
may optionally contain one or more olefinic bonds, unless otherwise constrained by the
definition Such cycloalkyl groups can include, for example, single ring structures, including
cyclopropyl, cyclobutyl, cyclooctyl, cyclopentenyl, and the like, or multiple ring structures,
including adamantanyl, and bicyclo [2 2 1] heptane, or cyclic alkyl groups to which is fused
an aryl group, for example, indane, and the like Spiro and fused ring structures can also be
included Cycloalkyl groups may be substituted further with one or more substituents
selected from alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino,
acyloxy, alkoxycarbonylamino, azido, cyano, halogen, hydroxy, oxo, thiocarbonyl, carboxy,
carboxyalkyl, arylthio, thiol, alkylthio, aryl, aralkyl, aryloxy, aminosulfonyl,
aminocarbonylamino, -NRfRq, -NHC (=O) NRfRq, -NHC (=O) Rf, -C (=O) NRfRq, -0-C
(=O)NRfRq (wherein Rf and Rq are the same as defined earlier), nitro, heterocyclyl,
heteroaryl, heterocyclylalkyl, heteroarylalkyl, or SO2-R60 (wherein R60 is same as defined
earlier) Unless otherwise constrained by the definition, cycloalkyl substituents optionally
may be substituted further by 1-3 substituents selected from alkyl, carboxy, hydroxy, alkoxy,
halogen, CF3, -NRfRq, -C(=O)NRfRq, -NHC(=O)NRfRq, -OC(=O)NRfRq (wherein Rf and Rq
are the same as defined earlier), cyano or -SO2R60 (where R60 is same as defined earlier)
"Cycloalkylalkyl" refers to alkyl-cycloalkyl group linked through alkyl portion, wherein the
alkyl and cycloalkyl are the same as defined earlier
The term "alkoxy" denotes the group O-alkyl, wherein alkyl is the same as defined
above
The term "aryl," unless otherwise specified, refers to carbocychc aromatic groups, for example, phenyl, biphenyl or napthyl ring and the like, optionally substituted with 1 to 3 substituents selected from halogen (eg, F, CI, Br, I), hydroxy, alkyl, alkenyl, alkynyl, cycloalkyl, alkoxy, acyl, aryloxy, CF3, cyano, nitro, COORe (wherein Re is hydrogen, alkyl, alkenyl, cycloalkyl, aralkyl, heterocyclylalkyl, heteroarylalkyl), NHC(=O)Rf, -NRfRq, -C(=O)NRfRq, -NHC(=O)NRfRq, -0-C(=O)NRfRq (wherein Rf and Rq are the same as defined earlier), -SO2R60 (wherein R60 is same as defined earlier), carboxy, heterocyclyl, heteroaryl, heterocyclylalkyl, heteroarylalkyl or amino carbonyl amino The aryl group optionally may be fused with a cycloalkyl group, wherein the cycloalkyl group may optionally contain heteroatoms selected from O, N or S
The term "aralkyl," unless otherwise specified, refers to alkyl-aryl linked through an alkyl portion (wherein alkyl is as defined above) and the alkyl portion contains 1-6 carbon atoms and aryl is as defined below Examples of aralkyl groups include benzyl, ethylphenyl and the like
The term "aralkenyl," unless otherwise specified, refers to alkenyl-aryl linked through alkenyl (wherein alkenyl is as defined above) portion and the alkenyl portion contains 1 to 6 carbon atoms and aryl is as defined below
The term "aryloxy" denotes the group O-aryl, wherein aryl is as defined above
The term "carboxy," as defined herein, refers to -C(=O)OH
The term "heteroaryl," unless otherwise specified, refers to an aromatic ring structure containing 5 or 6 ring atoms, or a bicychc aromatic group having from 8 to 10 ring atoms, with one or more heteroatom(s) independently selected from N, O or S optionally substituted with 1 to 4 substituent(s) selected from halogen (e g, F, CI, Br, I), hydroxy, alkyl, alkenyl, alkynyl, cycloalkyl, acyl, carboxy, aryl, alkoxy, aralkyl, cyano, nitro, heterocyclyl, heteroaryl, -NRfRq, CH=NOH, -(CH2)wC(=O)Rg {wherein w is an integer from 0-4 and Rg is hydrogen, hydroxy, ORf, NRfRq, -NHORz or -NHOH}, -C(=O)NRfRq and -NHC(=O)NRfRq , -SO2R6o -0-C(=O)NRfRq -0-C(=O)Rf, -0-C(=O)ORf (wherein R60, Rf and Rq are as defined earlier, and Rz is alkyl, cycloalkyl, aryl, heteroaryl, heterocyclyl, heteroarylalkyl or heterocyclylalkyl) Unless otherwise constrained by the definition, the substituents are attached to a ring atom, / e , carbon or heteroatom in the ring Examples of heteroaryl groups
include oxazolyl, lmidazolyl, pyrrolyl, 1,2,3-tnazolyl, 1,2,4-tnazolyl, tetrazolyl, thiazolyl, oxadiazolyl, benzoimidazolyl, thiadiazolyl, pyndinyl, pyndazinyl, pynmidinyl, pyrazinyl, thienyl, isoxazolyl, tnazinyl, fiiranyl, benzofuranyl, indolyl, benzothiazolyl, or benzoxazolyl, and the like
The term 'heterocyclyl," unless otherwise specified, refers to a non-aromatic monocyclic or bicychc cycloalkyl group having 5 to 10 atoms wherein 1 to 4 carbon atoms in a ring are replaced by heteroatoms selected from O, S or N, and optionally are benzofused or fused heteroaryl having 5-6 ring members and/or optionally are substituted, wherein the substituents are selected from halogen (eg, F, CI, Br, I), hydroxy, alkyl, alkenyl, alkynyl, cycloalkyl, acyl, aryl, alkoxy, alkaryl, cyano, nitro, oxo, carboxy, heterocyclyl, heteroaryl, -0-C(=O)Rf, -0-C(=O)ORf -C(=O)NRfRq, SO2R6o, -0-C(=O)NRfRq, -NHC(=O)NRq, -COORf, -NRfRq (wherein R60 Rf and Rq are as defined earlier) or guanidine Heterocyclyl can optionally include rings having one or more double bonds Unless otherwise constrained by the definition, the substituents are attached to the ring atom, / e , carbon or heteroatom in the ring Also, unless otherwise constrained by the definition, the heterocyclyl ring optionally may contain one or more olefinic bond(s) Examples of heterocyclyl groups include oxazohdinyl, tetrahydrofuranyl, dihydrofuranyl, dihydropyndinyl, dihydroisoxazolyl, dihydrobenzofuryl, azabicyclohexyl, dihydroindolyl, pyndinyl, isomdole 1,3-dione, pipendinyl, morphohnyl or piperazinyl
"Heteroarylalkyl" refers to alkyl-heteroaryl group linked through alkyl portion, wherein the alkyl and heteroaryl are as defined earlier
"Heterocyclylalkyl" refers to alkyl-heterocyclyl group linked through alkyl portion, wherein the alkyl and heterocyclyl are as defined earlier
"Acyl" refers to -C(=O)R" wherein R" is selected from hydrogen, alkyl, cycloalkyl, aryl, aralkyl, heteroaryl, heterocyclyl, heteroarylalkyl or heterocyclylalkyl
"Amine," unless otherwise specified, refers to -NH2 "Substituted amine," unless otherwise specified, refers to -N (Rk)2, wherein each Rk independently is selected from hydrogen {provided that both Rk groups are not hydrogen (defined as "ammo")}, alkyl, alkenyl, alkynyl, aralkyl, cycloalkyl, aryl, heteroaryl, heterocyclyl, heterocyclylalkyl, heteroarylalkyl, acyl, SO2R6o (wherein R60 is as defined above), -C(=O)NRfRq NHC(=O)NRfRq or -NHC(=O)ORf (wherein Rf and R, are as defined earlier)
"Thiocarbonyl" refers to -C(=S)H "Substituted thiocarbonyl" refers to-C(=S)R", wherein R" is selected from alkyl, cycloalkyl, aryl, aralkyl, heteroaryl, heterocyclyl, heteroarylalkyl or heterocyclylalkyl, amine or substituted amine
Unless otherwise constrained by the definition, all substituents optionally may be substituted further by 1-3 substituents selected from alkyl, aralkyl, cycloalkyl, aryl, heteroaryl, heterocyclyl, carboxy, carboxyalkyl, hydroxy, alkoxy, halogen, CF3, cyano, -C(=T)NRfRq, -0(C=0)NRfRq (wherein Rf, Rq and T are the same as defined earlier) and -OC(=T)NRfRq, -SO2R6o (where R60 is the same as defined earlier)
The term "pharmaceutically acceptable salts" refers to derivatives of compounds that can be modified by forming their corresponding acid or base salts Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acids salts of basic residues (such as amines), or alkali or organic salts of acidic residues (such as carboxylic acids), and the like
The term "peptide" refers to a molecule comprising a series of amino acids linked through amide linkages Dipeptides comprise 2 amino acids, tnpeptides comprise 3 amino acids and tetrapeptides comprise four amino acids, wherein the term amino acid is as defined earlier
The present disclosure includes all isotopes of atoms occurring in the present compounds Isotopes include those atoms having the same atomic number but different mass numbers By way of general example, and without limitation, isotopes of hydrogen include tritium and deuterium Isotopes of carbon include C-13 and C-14
The compounds provided herein can contain one or more asymmetric carbon atoms and may thus occur as racemates and racemic mixtures, single enantiomers, diastereomeric mixtures and individual diastereomers All such isomeric forms of these compounds are expressly included in the present invention Each stereogenic carbon may be of the R or S configuration Although specific compounds may be depicted in a particular stereochemical configuration, compounds having either the opposite stereochemistry at any given chiral center or mixtures thereof are envisioned as part of the invention Although amino acids and amino acid side chains may be depicted in a particular configuration, both natural and unnatural forms are envisioned
Detailed description of the invention
The compounds disclosed herein may be prepared by techniques well known in the art and familiar to the ordinarily skilled synthetic organic chemist In addition, the compounds provided herein may be prepared by, for example, the following reaction sequences, for example as depicted in Schemes I, II, III and IV
(Scheme Removed)
Compounds of Formula XIII can be prepared by following the Scheme I Accordingly, compounds of Formula II are reacted with compounds of Formula III to give compounds of Formula IV (wherein R6 can be alkyl), which on coupling with compounds of Formula V give compounds of Formula VI (wherein ring A, X can be the same as defined earlier and P can be a protecting group), which on reduction give compounds of Formula VII, which on coupling with compounds of Formula VIII (wherein R7 can be hydroxy or halogen) give compounds of Formula IX ( wherein R3 can be the same as defined earlier), which on deprotection give compounds of Formula X, which on condensation with compounds of Formula XI give compounds of Formula XII ( wherein Rj can be the same as defined earlier), which is finally hydrolysed to give compounds of Formula XIII Compounds of
Formula XIII can be further, converted to pharmaceutically acceptable salts using the methods well known to one ordinary skilled in art
The reaction of compounds of Formula II with an alcohol of Formula III, for example, ethanol or methanol to give compounds of Formula IV can be carried in presence of an acid, for example, sulfuric acid and hydrochloric acid
The coupling of compounds of Formula IV with compounds of Formula V to give compounds of Formula VI can be carried out in dimethylformamide, dichloromethane or tetrahydrofuran, with a coupling agent, for example, l-(3-dimethylamino propyl)-3-ethyl-carbodimide or dicyclohexylcarbodnmide in the presence of 1-hydroxbenzotnazole, and a base, such as N-methylmorphohne or tnethylamine
The reduction of compounds of Formula VI to give compounds of Formula VII can be carried out in an atmosphere of hydrogen in the presence of a catalyst, for example, palladium on carbon or raney nickel in solvents, for example, methanol, ethanol, isopropanol or ethyl acetate
The coupling of compounds of Formula VII with compounds of Formula VIII to give compounds of Formula IX can be carried out in dimethylformamide, dichloromethane or tetrahydrofuran, with or without a coupling agent, for example, l-(3-dimethylamino propyl)-3-ethyl-carbodimide or dicyclohexylcarbodnmide in the presence of 1-hydroxbenzotnazole in presence of a base, for example, N-methylmorphohne or tnethylamine
The deprotection of compounds of Formula IX to give compounds of Formula X can be earned out in the presence of reagents, for example, tnfluoroacetic acid or hydrochloric acid in an organic solvent, for example, dichloromethane, ethyl acetate or dioxane
The condensation of compounds of Formula X with compounds of Formula XI to give compounds of Formula XII can be carried out m an organic solvent, for example, tetrahydrofuran, dichloromethane, dimethylformamide or acetomtnle
The hydrolysis of compounds of Formula XII to give compounds of Formula XIII can be carried out in a solvent, for example, aqueous tetrahydrofuran, aqueous methanol or aqueous ethanol and combination(s) thereof, in the presence of a base, for example, lithium hydroxide, sodium hydroxide or potassium hydroxide
Illustrative compounds prepared following Scheme I include
- (S)-3-{4-[(Pyndine-4-carbonyl)-amino]-phenyl}-2-[((S)-l-p-
tolylcarbamoylpyrrohdine2-carbonyl)-amino]-propiomc acid (Compound No 1)
and its pharmaceutically acceptable salts, pharmaceutically acceptable solvates, stereoisomers, tautomers, racemates, regioisomers, prodrugs, metabolites, polymorphs or N-oxides
(Scheme Removed)
Compounds of Formula XXIV and XXVII can be prepared by following Scheme II Accordingly, compounds of Formula XIV on reaction with compounds of Formula XV (wherein X can be halogen) give compounds of Formula XVI (wherein R\ can be the same as defined earlier), which on condensation with compounds of Formula XVII give compounds of Formula XVIII (wherein ring A, R6 and X can be the same as defined earlier), which on hydrolysis give compounds of Formula XIX, which on coupling with compounds of Formula IV give compounds of Formula XX, which on reduction give compounds of Formula XXI,
(a) which on reaction with compounds of Formula XXII give compounds of Formula
XXIII (wherein R4 can be the same as defined earlier), which is finally
hydrolyzed to give compounds of Formula XXIV
(b) which on reaction with compounds of Formula XXV give compounds of Formula
XXVI (wherein R3 can be same as defined earlier), which is finally hydrolyzed to
give compounds of Formula XXVII
Compounds of Formula XXIV and XXVII can be further, converted to pharmaceutically acceptable salts using the methods well known to one ordinary skilled in art
The reaction of compounds of Formula XIV with compounds of Formula XV to give compounds of Formula XVI can be carried out in an organic solvent, for example, tetrahydrofuran, dichloromethane, dimethylformamide or acetonitnle in the presence of an inorganic base, for example, sodium bicarbonate or potassium bicarbonate
The condensation of compounds of Formula XVI with compounds of Formula XVII to give compounds of Formula XVIII can be carried out in an organic solvent, for example, tetrahydrofuran, dimethylformamide or dioxane in presence of an organic base such as tnethylamine, dnsopropylethylamine or pyridine
The hydrolysis of compounds of Formula XVIII to give compounds of Formula XIX can be carried out in a solvent, for example, aqueous tetrahydrofuran, aqueous methanol or aqueous ethanol and combination(s) thereof, in the presence of a base, for example, lithium hydroxide, sodium hydroxide or potassium hydroxide
The coupling of compounds of Formula XIX with compounds of Formula IV to give compounds of Formula XX can be carried out in dimethylformamide, dichloromethane or tetrahydrofuran, with a coupling agent, for example, l-(3-dimethylammo propyl)-3-ethyl-carbodimide or dicyclohexylcarbodnmide in the presence of 1-hydroxbenzotnazole, and a base, such as N-methylmorphohne or tnethylamine
The reduction of compounds of Formula XX to give compounds of Formula XXI can be carried out in an atmosphere of hydrogen in the presence of a catalyst, for example, palladium on carbon or raney nickel in solvents, for example, methanol, ethanol, isopropanol or ethyl acetate
The reaction of compounds of Formula XXI with compounds of Formula XXII or compounds of Formula XXV to give compounds of Formula XXIII or compounds of Formula XXVI respectively can be carried out in an organic solvent, for example, tetrahydrofuran, dichloromethane, dimethylformamide or acetonitnle in the presence of an organic base such as tnethylamine, dnsopropylethylamine or pyridine
The hydrolysis of compounds of Formula XXIII or compounds of Formula XXVI to give compounds of Formula XXIV or compounds of Formula XXVII respectively can be carried out in a solvent, for example, aqueous tetrahydrofuran, aqueous methanol or aqueous ethanol and combination(s) thereof, in the presence of a base, for example, lithium hydroxide, sodium hydroxide or potassium hydroxide
Illustrative compounds prepared following Scheme II include
- (S)-3-(4-Acetylamino-phenyl)-2- {[ 1 -(biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-propionic acid (Compound No 2)
(S)-2-{[l-Biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-3-[4-(2,2,2-tnfluoro-acetylamino)-phenyl] -propionic acid (Compound No 3)
(S)-2-{[l-(Biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-3-(4-methanesulfonyl amino-phenyl)-propionic acid (Compound No 4)
(S)-3-(4-Benzenesulfonylamino-phenyl)-2-{[l-(biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-propionic acid (Compound No 5)
(S)-2-{[l-(Biphenyl-2-ylcarbamoyl)-pyrrolidme-2-carbonyl]-amino}-3-(4-toluene-4-sulfonyl amino) phenyl)-propiomc acid (Compound No 6)
and their pharmaceutically acceptable salts, pharmaceutically acceptable solvates,
stereoisomers, tautomers, racemates, regioisomers, prodrugs, metabolites, polymorphs or N-
oxides
(Scheme Removed)
Compounds of Formula XIII, XXXI and XXXII can be prepared by following Scheme III Accordingly, compounds of Formula VI, on deprotection give compounds of
Formula XXVIII (wherein ring A, R6 and X can be the same as defined earlier), which on condensation with compounds of Formula XI or compounds of Formula XVI give compounds of Formula XXIX,
(a) which on reduction give compounds of Formula XXX, which on coupling with compounds of Formula VIII ( wherein R7 can be the same as defined earlier) give compounds of Formula XII (wherein R3 can be the same as defined earlier), which is finally hydrolysed to give compounds of Formula XIII
(b) which on hydrolysis give compounds of Formula XXXI, which is finally reduced to give compounds of Formula XXXII
Compounds of Formula XIII, XXXI and XXXII can be further, converted to pharmaceutically acceptable salts using the methods well known to one ordinary skilled in art
The deprotection of compounds of Formula VI to give compounds of Formula XXVIII can be carried out in the presence of reagents, for example, trifluoroacetic acid, or hydrochloric acid in an organic solvent, for example, dichloromethane, ethyl acetate or dioxane
The condensation of compounds of Formula XXVIII with compounds of Formula XI or compounds of Formula XVI to give compounds of Formula XXIX can be carried out in an organic solvent, for example, tetrahydrofuran, dimethylformamide or dioxane in presence of an organic base such as triethylamine, dnsopropylethylamine or pyridine
The reduction of compounds of Formula XXIX or compounds of Formula XXXI to give compounds of Formula XXX or compounds of Formula XXXII respectively can be carried out in an atmosphere of hydrogen, in the presence of a catalyst, for example, palladium on carbon or raney nickel in solvents, for example, methanol, ethanol, isopropanol or ethyl acetate
The coupling of compounds of formula XXX with the compounds of Formula VIII to give compounds of Formula XII can be carried out in dimethylformamide, dichloromethane or tetrahydrofuran, with a coupling agent, for example, l-(3-dimethylamino propyl)-3-ethyl-carbodimide or dicyclohexylcarbodumide in the presence of 1-hydroxbenzotnazole, and a base, such as N-methylmorpholine or triethylamine
The hydrolysis of compounds of Formula XII or compounds of Formula XXIX to give compounds of Formula XIII or compounds of Formula XXXI respectively can be carried out in a solvent, for example, aqueous tetrahydrofuran, aqueous methanol or aqueous ethanol and combination(s) thereof, in the presence of a base, for example, lithium hydroxide, sodium hydroxide or potassium hydroxide
Illustrative compounds prepared following Scheme III include
- (S)-3-{4-[(Pyndine-4-carbonyl)-amino]-phenyl}-2-[((S)-l-o-tolylcarbamoyl-
pyrrohdine-2-carbonyl)-amino]-propionic acid (Compound No 7)
(S)-2-{[S-l-(2-Methoxy-phenylcarbamoyl)-pyrrolidine-2-carbonyl]-ammo}-3-{4-[(pyndine-4-carbonyl)-amino]-propionic acid (Compound No 8)
(S)-2-({(S)-l-[(Benzo[l,3]dioxol-5-ylmethyl)-carbamoyl]-pyrrohdine-2-carbonyl}-amino)-3{4-[(pyndine-4-carbonyl)-amino]-phenyl}-propionic acid (Compound No 9)
(S)-2-{[(S)-l-(Biphenyl-2-ylcarbamoyl)-pyrrohdme-2-carbonyl]-amino}-3-{4-[(pyndine-4-carbonyl)-amino]-phenyl}-propiomc acid (Compound No 10)
(S)-3-(4-Benzoylammo-phenyl)-2-{ [ 1 -(S)-(biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-propionic acid (Compound No 11)
- 4-[4-((S)-2-{[(S)-l-(Biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-2-
carboxy-ethyl)-phenylcarbamoyl]-pipendine-l-carboxyhc acid tert-butyl ester
(Compound No 12)
(S)-2-{[(S)-l-(Biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-3-(4-nitro-phenyl)-propiomc acid (Compound No 14)
(S)-3-(4-Amino-phenyl)-2-{[(S)-l-(biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino} propionic acid (Compound No 15)
- 4-{4-[(S)-2-({(S)-l-[(Benzo[l,3]dioxol-5-ylmethyl)carbamoyl]-pyrrohdine-2-
carbonyl}-amino)-2-carboxy-ethyl]-phenylcarbamoyl}-pipendine-l-carboxylic acid
tert-butyl ester (Compound 16)
and their pharmaceutically acceptable salts, pharmaceutically acceptable solvates,
stereoisomers, tautomers, racemates, regioisomers, prodrugs, metabolites,
polymorphs or N-oxides,
- (S)-2-{[(S)-l-(Biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-3-{4-
[(pipendine-4-carbonyl)-amino]-phenyl}-propionic acid TFA salt (Compound No
13)
and its pharmaceutically acceptable solvates, stereoisomers, tautomers, racemates,
regioisomers, prodrugs, metabolites, polymorphs or N-oxides
(Scheme Removed)
Compounds of Formula XXXV and XXXVIII can be prepared by following Scheme IV Accordingly, the coupling of compounds of Formula XIX with compounds of Formula XXXIII gives compounds of Formula XXXIV (wherein ring A, X, Ri, R2 and R6 can be the same as defined earlier)
(a) which, is finally hydrolyzed to give compounds of Formula XXXV
(b) which on reaction with compounds of Formula XXXVI ( wherein Xi can be halogen) give compounds of Formula XXXVII (wherein R5 can be the same as defined earlier), which is finally hydrolyzed to give compounds of Formula XXXVIII
Compounds of Formula XXXV and XXXVIII can be further, converted to pharmaceutically acceptable salts using the methods well known to one ordinary skilled in art
The coupling of compounds of Formula XIX with compounds of Formula XXXIII to give compounds of Formula XXXIV can be carried out in dimethylformamide, dichloromethane or tetrahydrofuran, with a coupling agent, for example, l-(3-dimethylamino propyl)-3-ethyl-carbodimide or dicyclohexylcarbodnmide in the presence of 1-hydroxbenzotnazole, and a base, such as N-methylmorphohne or tnethylamine
The reaction of compounds of Formula XXXIV with the compounds of Formula XXXVI to give compounds of Formula XXXVII can be carried out inorganic solvent, for example, dimethylformamide, dichloromethane or tetrahydrofuran in the presence of inorganic base, for example, potassium carbonate or sodium carbonate
The hydrolysis of compounds of Formula XXXIV or compounds of Formula XXXVII to give compounds of Formula XXXV and compounds of Formula XXXVIII respectively can be carried out in a solvent, for example, aqueous tetrahydrofuran, aqueous methanol or aqueous ethanol and combination(s) thereof, in the presence of a base, for example, lithium hydroxide, sodium hydroxide or potassium hydroxide
Illustrative compounds prepared following Scheme IV include
- (S)-2-{[l-(Biphenyl-2-ylcarbamoyl)-pyrrolidine-2-carbonyl]-ammo}-3-(4-fluoro-
phenyl)-propionic acid (Compound No 17)
- (S)-2-{[l-(Biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-3-(4-hydroxy-phenyl)-propiomc acid (Compound No 18)
- (S)-2- {[ 1 -(Biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino } -3-[4-(2-morpholin-4yl-ethoxy)-phenyl}-propiomc acid (Compound No 19)
and their pharmaceutically acceptable salts, pharmaceutically acceptable solvates,
stereoisomers, tautomers, racemates, regioisomers, prodrugs, metabolites, polymorphs or N-
oxides
The present disclosure includes all isotopes of atoms occurring in the present compounds Isotopes include those atoms having the same atomic number but different mass numbers By way of general example, and without limitation, isotopes of hydrogen include tritium and deuterium Isotopes of carbon include C-13 and C-14
The compounds provided herein can contain one or more asymmetric carbon atoms and may thus occur as racemates and racemic mixtures, single enantiomers, diastereomenc mixtures and individual diastereomers All such isomeric forms of these compounds are expressly included in the present invention Each stereogenic carbon may be of the R or S configuration Although specific compounds may be depicted in a particular stereochemical configuration, compounds having either the opposite stereochemistry at any given chiral center or mixtures thereof are envisioned as part of the invention Although amino acids and amino acid side chains may be depicted in a particular configuration, both natural and unnatural forms are envisioned
The compounds disclosed herein may be prepared by techniques well known in the art and familiar to the skilled synthetic organic chemist
Where desired, the compounds of Formula I and/ or their pharmaceutically acceptable salts, pharmaceutically acceptable solvates, stereoisomers, tautomers, racemates, regioisomers, prodrugs, metabolites, polymorphs or N-oxides may be advantageously used in combination with one or more other therapeutic agents Examples of other therapeutic agents, which may be used in combination with compounds of Formula I of this invention and/ or their pharmaceutically acceptable salts, pharmaceutically acceptable solvates, stereoisomers, tautomers, racemates, regioisomers, prodrugs, metabolites, polymorphs or N-oxides include corticosteroids, beta agonists, leukotnene antagonists, 5-hpoxygenase inhibitors, phosphodiesterase inhibitors, chemokme inhibitors and muscarinic receptor antagonists
Examples set forth below demonstrate the general synthetic procedure for the preparation of representative compounds The examples are provided to illustrate particular aspect of the disclosure and do not limit the scope of the present invention
Experimental details
Example 1 Preparation of (S)-2-Amino-3-(4-nitrophenyl)-propiomc acid methyl ester hydrochloride
Methanolic hydrochloric acid (4N, 100 ml) under stirring was added to (S)-4-
Nitrophenyl-alanine (16, l0g) at room temperature and was stirred for 18h and concentrated
in vacuum to obtain the title compound as light yellow solid Yield 5 6g, (100%)
IR (vmax, DCM) 3463, 2915, 1744, 1606, 1350, 1241, 1108, 751 cm ', 1HNMR (DMSO-d6)
δ 3 16-3 27(2H, m, ArCH2O, 3 68 (3H, s, OCH3), 4 35-4 40 (1H, m, NHCHCO), 7 62 (2H, d, J = 9Hz) and 8 20 (2H, d, J = 9 Hz) [aromatic], 8 78 (2H, bs, NH2), MS (m/z) 225 2 (M+l)
Example 2 Preparation of (S)-2-[(S)-l-Methoxycarbonyl-2-(4-nitrophenyl)-
ethylcarbamoyl]-pyrrolidine-l-carboxyhc acid tert-butyl ester
N-methyl morphohne (6 9g, 69 mmol), 1-hydroxybenzotnazole (3 4g, 25 mmol) and
Boc-Prohne (5g, 23 mmol) were added to a solution of amine (example 1, 6g, 23 mmol) in
dimethylformamide (25 ml) at 0°C Reaction mixture was stirred for about 30 minutes and 1-
(3-dimethylaminopropyl)-3-ethyl-carbodimide hydrochloride (4 9g, 25mmol) was added
maintaining same temperature Reaction mixture was stirred at room temperature for
overnight, diluted with water (20 ml) and extracted with ethyl acetate (2 x 50 ml) Combined
organic phase was washed with water (2 x 20 ml), saturated aqueous sodium bicarbonate (20
ml) and brine (2 x 20 ml) Organic phase was dried (anhydrous sodium sulphate) and
concentrated in vacuum to afford the crude product Purification by column chromatography (Silica gel, 100-200mesh, 50 50, ethyl acetate hexane) afforded the title compound as sticky solid Yield 9 2g(94%)
1H NMR (CDCI3) δ 1 43 (9H, s, C(CH3)3), 1 38-2 00 (3H, m) and 2 23-2 36 (1H, m) [2 X CH2 -CH2], 2 88-2 96 (2H, m, Ar CH2), 3 08-3 15 (2H, NHCH2), 3 74 (3H, s, OCH3), 4 24-4 26 (1H, m, NCHCO), 4 87-4 89 (1H, m, NHCHCO), 7 27-7 31 (3H, m) and 8 13-8 15 (2H, d, J = 9Hz) [aromatic and NH], MS (m/z) 422 2 (M+l)
Example 3 (S)-2-r(S)-2-(4-Amino-phenyl)-1 -methoxycarbonyl-ethylcarbamoyl]-pyrrohdine-1-carboxyhc acid tert-butyl ester
Dry palladium on carbon (10%, 200 mg) was added to a solution of nitro compound (example 2, 1 8g, 4 2 mmol) in methanol (20 ml) Hydrogenation was carried out using a Pan-apparatus at 50 psi for about 1 hr and the mixture was filtered through cehte pad Filtrate was concentrated to obtain the title compound as sticky mass Yield 1 5 g (90%)
IR (vmax, DCM) 3428, 1744,1669, 1518, 1402, 1366, 1281, 1256, 1163, 1124, 853, 734 cm ', MS (m/z) 392 4 (M+l)
Example 4 Preparation of (S)-2-((S)-l-Methoxycarbonyl-2{4-[(pyndine-4-carbonyl)-amino]-phenyl}-ethylcarbamoyl)-pyrrohdine-l-carboxyhc acid tert-butyl ester
Isomcotinic acid (440 mg, 3 5 mmol), 1-hydroxybenzotriazole (601 mg, 3 93 mmol)
and N-methyl morphohne (920 mg, 3 93 mmol) were added to a solution of amine (example
3, 200 mg, 0 47 mmol) in dimethylformamide (5ml) at about 0°C Reaction mixture was
stirred for about 30 minutes at 0°C and l-(3-dimethylamino propyl)-3-ethyl-carbodimide
hydrochloride (99 mg, 0 52 mmol) was added Reaction mixture was then stirred at room
temperature for overnight and diluted with ethyl acetate (20 ml), washed with water (2x5
ml) and brine (2 x 15 ml) Organic phase was dried (anhydrous sodium sulphate) and
concentrated under reduced pressure to obtain crude product Purification by column
chromatography (silica gel, 100-200 mesh, 3 97, methanol DCM) afforded the title
compound as hygroscopic solid Yield 1 5g (84%)
IR (vmax, DCM) 3533, 3300, 3051, 2979, 2881, 1750, 1696, 1665, 1642, 1605, 1534, 1437, 1413, 1386, 1325, 1271, 1211, 1157, 1123 cm1, 1H NMR (Acetone-d6) δ 142 (9H, s, C(CH3)3), 1 81-1 83 (2H, m), 1 92-2 01 (2H, m) [CH2CH2], 3 00-3 11 (1H, m) and 3 14-3 21 (1H, m) [ArCH2], 3 35-3 48 (2H, m, NHCH2), 3 73 (3H, s, OCH3), 4 24 (1H, bs, NCHCO), 4 84-4 85 (1H, NHCHCO), 7 12-7 16 (2H, m), 7 56-7 59 (2H, m), 7 71-7 72 (2H, m), 8 0-8 1 (1H, m), 8 79 (2H, d, J = 6Hz) [8H aromatic +1NH], MS (m/z) 497 3 (M+l)
Example 5 (S)-3-{4-rCPyridine-4-carbonyl)-amino]phenvl}-2-[((S)-pyrrohdine-2-carbonyl)-amino]-propiomc acid methyl ester
Tnfluoroacetic acid (6 ml) was added to a solution of Boc compound (example 4, 1 5
g, 3 02 mmol) in dichloromethane (30 ml) at room temperature, the reaction mixture was
stirred for about 2h and concentrated in vacuum to obtain crude mass that was dissolved in
acetone (20 ml) and potassium carbonate (lg) was added Reaction mixture was stirred for
about 10 minutes and filtered Filtrate was concentrated in vacuum to obtain the title
compound as light yellow solid (very hygroscopic) Yield 1 2 g (100%)
1H NMR (Acetone-d6) 6 1 57-1 60 (1H, m), 1 72-1 73 (1H, m) and 1 90-2 10 (2H, m) [CH2CH2], 2 66-2,77 (1H, m) and 2 82-2 85 (1H, m) [ArCH2,], 3 37-3 38 (1H, m) and 3 62-3 63 (1H, m) [NHCH23], 3 71 (3H, s, OCH3), 4 19-4 24 (1H, m, NCHCO), 4 72-4 73 (1H, m, NHCHCO), 7 18 (1H, d, J = 8Hz), 7 27 (1H, d, J = 8Hz), 7 79 (2H, d, J = 8Hz), 7 89 (2H, d, J = 6Hz), 8 75 (2H, d, J = 6Hz) [8H aromatic], 9 90 (1H, s, NH), MS (m/z) 397 3 (M+l)
Example 6 Preparation of (S)-3-{4-[(Pyndine-4-carbonyl)-amino]-phenyl}-2-[((S)-l-p-tolylcarbamoyl-pyrrohdine-2-carbonyl)-armnol -propionic acid methyl ester
4-Tolyl isocyanate (73 mg, 0 55 mmol) was added to a solution of amine (example 5,
220 mg, 0 55 mmol) in acetomtrile (5ml) at about 0°C and stirred at same temperature for
about 30 minutes Reaction mixture was diluted with ethyl acetate (10 ml), washed with
water and brine (10 ml each), dried (anhydrous sodium sulphate) and concentrated to obtain
crude product Purification by column chromatography (silica gel, 100-200 mesh, 4 96,
Methanol dichloromethane) afforded the title compound as sticky mass Yield 190 mg
(64%)
IR(vmax, DCM) 3492, 2924, 1738, 1661, 1600, 1532, 1443, 1411, 1353, 1327, 1243, 1118, 1018, 817, 753, 673 cm 1 1HNMR (Acetone-d6) δ 2 13-2 51 (4H, m, CH2CH2), 2 78 (3H, s, ArCH3), 2 81-2 97 (1H, m) and 3 01-3 11 (1H, m) [ArCHJ, 3 43-3 52 (2H, m, NHCH2), 3 69 (3H, s, OCH3), 4 45-4 48 (1H, m, NCHCO), 4 75-4 76 (1H, m, NHCHCO), 7 03 (2H, d, J = 8Hz), 7 20 (2H, d, J = 8Hz), 7 45 (2H, d, J = 9Hz), 7 54 (1H, s), 7 65-7 68 (2H, m), 7 83-8 75 (3H, m), 8 76-8 77 (2H, m), 9 64 (1H, s) [12 H aromatic + 3 X NH], MS (m/z) δ30 2 (M+l)
Example 7 Preparation of (SV-3-{4-[(Pyndine-4-carbonyl)-amino]-phenyl}-2-r((S)-l-p-tolylcarbamoyl-pyrrohdine-2-carbonyl)-amino]-propionic acid (Compound No 1)
Lithium hydroxide (8 mg, 0 18 mmol) was added to a solution of methyl ester
(example 6, l00mg, 0 18 mmol) in tetrahydrofuran (3 ml), methanol (1 ml) and water (1 ml)
Reaction mixture was stirred for about 3 hr and concentrated to obtain the crude oil, which
was taken in water (10 ml) and extracted with ethyl acetate (2x10 ml) Aqueous layer was
acidified with sodium hydrogensulfate to pH 3 and extracted with ethyl acetate (2x10 ml) This combined organic phase was washed with water and brine (10 ml each), dried (anhydrous sodium suphate) and concentrated to obtain the desired compound as sticky mass Yield 68 mg (70%) (Mixture of diastereomers, 96 2 by HPLC, tR = 18 50 & 22 21 mm , PDA 233 0 nm)
1H NMR (DMSO-d6) δ 1 81-1 97 (4H, m, CH2CH2), 2 18 (3H, s, ArCH3), 2 94-3 02 (2H, m, ArCH2), 2 90-3 09 (2H, m, ArCH2), 3 35-3 51 (2H, m, NHCH2), 4 33-4 36 (2H, m, NHCHCO and NCHCO), 7 02 (2H, d, J = 8Hz), 7 20 (2H, d, J = 8Hz), 7 31 (2H, d, J = 9Hz), 7 61 (2H, d, J = 9Hz), 7 83-7 85 (2H, m), 8 76-8 78 (2H, m) [aromatic], MS (m/z) δ16 2 (M+l)
Example 8 Preparation of (S)-3-(4-Nitro-phenyl)-2-[((S)-pyrrohdine-2-carbonyl)-amino]-propionic acid methyl ester
Boc amine (example 2, lg, 2 3 mmol) was stirred with tnfluoroacetic acid (6 ml) in
dichloromethane (14 ml) at room temperature for about 2h and concentrated under reduced
pressure to obtain the deprotected amine tnfluoroacetate salt as oil, which was then taken in
acetone and potassium carbonate (lg) was added Reaction mixture was stirred for about 5
minutes and filtered through cehte pad Filtrate was concentrated to obtain the yellow oil,
which was used without any purification
Example 9 Preparation of (S)-3-(4-Nitrophenyl)-2-[(l-o-tolylcarbamoyl-pyrrohdine-2-carbonyl) amino]-propionic acid methyl ester
o-Tolyl isocyanate (315mg, 2 3 mmol) was added to a solution of amine (example 8,
crude) in tetrahydrofuran Reaction mixture was stirred for about 30 minutes at about 0°C,
followed by stirring at room temperature for overnight Reaction mixture was then heated at
about 60°C for about 5 h till completion of reaction Reaction mixture was diluted with ethyl
acetate (20 ml), washed with water and brine (15 ml each) Organic layer was dried
(anhydrous sodium sulphate) and concentrated in vacuum to obtain crude product
Purification by column chromatography (silica gel, 100-200 mesh, 3 97 methanol
dichloromethane) afforded the title compound as sticky mass Yield 1 lg (88 %)
IR (vmax, DCM) 3410, 2953, 1742, 1644, 1520, 1453, 1346, 1252, 1112, 753 cm 1 1HNMR (CDCI3) δ 1 87-2 00 (3H, m) and 2 36-2 24 (1H, m) [2 X CH2 -CH2], 2 23 (3H, s, ArCHj), 3 02-3 10 (1H, m) and 3 27-3 37 (3H, m) [ArCHj. and NCH2), 3 72 (3H, m, OCH3) 4 52-4 55 (1H, m, NCHCO), 4 83-4 86 (1H, m, NHCHCO), 6 09 (1H, bs, NH), 7 03-7 08 (1H, m), 7 19-7 30 (4H, m), 7 75-7 80 (2H, m) and 7 95-7 97 (2H, m) [aromatic + NH), MS (m/z) 455 2 (M+l)
Example 10 Preparation of (S)-2-{[(S)-l-(2-Methoxy-phenylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-3-(4-nitro-phenyl)-propionic acid methyl ester
Title compound was prepared via same method as used in example 9 using amine
(example 8, crude) and 2-methoxyphenyl isocyanate (354 mg, 2 3 mmol) Purification by
column chromatography (silica gel, 100-200 mesh, 3 97, methanol DCM) afforded the title
compound as sticky mass Yield (810 mg, 74%)
IR (vmax, DCM) 3425, 1744, 1651, 1522, 1459, 1346, 1249, 1026, 751 cm 1 1HNMR (DMSO-d6) δ 1 73-1 85 (3H, m) and 1 99-2 05 (1H, m) [2 X CH2 -CHJ, 3 11-3 30 (2H, m, A1CH2) 3 38-3 52 (2H, m, NCH2), 3 64 (3H, s, ArOCHs), 3 81 (3H, s, OCH3), 4 29-4 31 (1H, m, NCHCO), 4 57-4 61 (1H, m, NHCHCO), 6 85-6 89 (1H, m), 6 94-7 01 (2H, m), 7 16 (1H, s), 7 51 (2H, d, J = 8Hz), 7 88 (1H, d, J= 7Hz), 8 06 (2H, d, J= 8Hz), 8 42 (1H, d, J= 8Hz) [8H aromatic + 2 NH], MS (m/z) 471 3 (M+l)
Example 11 Preparation of Biphenyl-2-yl-carbamic acid 4-mtro-phenyl ester
Sodium bicarbonate (496 mg, 5 9 mmol) followed by 4-mtro-phenylchloroformate (1 19g, 5 9 mmol) was added to cold (0°C) solution of 2-aminobiphenyl (lg, 5 9 mmol) in tetrahydrofuran (10 ml) Reaction mixture was stirred at same temperature for about 30 minutes and at room temperature for about 1 hr Reaction mixture was diluted with ethyl acetate (20 ml) and washed with water (2x10 ml), dried over anhydrous sodium sulphate and concentrated to obtain the oily compound which was stirred with hexane (20 ml) and filtered to obtain crude product as brown solid which was used as such Yield 1 6g (crude)
Example 12 Preparation of Benzo[l,3]dioxol-5-ylmethyl-carbamic acid-4-nitro-methyl ester Piperanyl amine (lg, 6 6 mmol) was reacted with 4-nitrophenyl chloroformate (1 33g, 6 6 mmol) via same procedure as in example 11 to obtain the title compound, which was used further without any purification Yield 1 3g (crude)
Example 13 Preparation of (S)-2-{[(S)-l-(Biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-3-(4-mtro-phenyl)-propionic acid methyl ester
Ethyldnsopropylamine (612 mg, 2 3 mmol) followed by 4-nitrophenyl carbamate
(1 58g, 4 6 mmol) was added to a solution of compound of example 8 (4 lg crude) in
tetrahydrofuran at 0°C and stirred at same temperature for about 30 minutes and at room
temperature for overnight Reaction mixture was diluted with ethyl acetate (20 ml) and
washed with water and brine (15 ml each) Organic phase was dried (anhydrous sodium
sulphate) and concentrated to obtain the crude compound Purification by column

chromatography (silica gel, 100-200 mesh, 40 60, ethyl acetate hexane) afforded the title compound as sticky mass Yield 1 6 g (66%)
IR (vraax, DCM) 3426,3057,2953, 1743, 1673, 1521, 1446, 1347, 1272, 1215, 1112,857, 737, 703cm 1 1H NMR (DMSO-d6) δ1 68-1 98 (4H, m, CH2CH2), 3 03-3 24 (4H, m, NCHJJ and ArCH:), 3 60 (3H, s, OCH3) 4 17-4 19 (1H, m, NCHCO), 4 53-4 54 (1H, m, NHCHCO), 7 18-751 (10H, m), 7 68 (1H, d, J = 9Hz) and 8 08 (2H, d, J = 9Hz) [aromatic] 8 30 (1H, d, NH),MS(m/z) δ17 3(M+1)
Example 14 Preparation of (S)-2-({(S)-l[(Benzo[l 3]dioxol-5-ylmethyl)-carbamoyl]-pyrrohhdine-2-carbonyl}-amino)-3-(4-nitrophenyl)-propionic acid methyl ester
Amine (example 8 crude) and 4-nitrophenyl carbamate (example 11,0 75g, crude)
were reacted via same procedure as for compound in example 13 to obtain title compound as
crude yellow oil Purification by column chromatography (silica gel, 100 - 200 mesh, 60 40
ethyl acetate hexane) afforded the title compound as sticky mass Yield 920 mg (78%)
1H NMR (CDCI3) δ 1 80-1 93 (3H, m) and 2 32-2 34 (1H, m) [CHaCHJ, 3 04-3 35 (4H, m, ArCH2 and NHCH2), 3 74 (3H, s, OCH3), 4 44-4 47 (2H, m, NHQfcAr), 4 67-4 68 (1H, m, NCHCO), 4 84-4 86 (1H, m, NHCHCO), 5 93 (2H, s, OCH2O), 6 76-6 81 (3H, m), 7 26-7 30 (3H, m), 7 89 (1H, m, J = 8Hz) δ 07 (2H, d, J = 9Hz) [7H aromatic and 2H NH], MS (m/z) 499 2 (M+l)
Example 15 Preparation of (S)-3-(4-Amino-phenyl)-2-[((S)-l-o-tolylcarbamoyl-pyrrohdine-2-carbonyl)-amino]-propionic acid methyl ester
Dry 10% palladium on charcoal (200 mg) was added to a solution of nitro compound
(example 9, 850 mg, 1 8 mmol) in methanol (20 ml) Reaction mixture was shaken under
hydrogen atmosphere at 50 psi for about 1 hr and filtered through cehte pad Filtrate was
concentrated to obtain the desired compound as sticky mass Yield 680 mg (86 %)
IR(vmax,DCM) 3358,2951, 1742, 1656, 1519, 1451, 1364, 1251, 1119,755 cm1 ]HNMR (CDCI3) δ 1 94-1 99 (3H, m) and 2 34-2 35 (1H, m) [2 X CH2 -CH2], 2 24 (3H, s, ArCH3), 2 88-2 95 (1H, m) and 3 03-3 09 (1H, m) [ArCHJ, 3 40-3 48 (2H, m, NCH2), 3 69 (3H, s, OCH3) 4 49-4 51 (1H, m, NCHCO), 4 73-4 75 (1H, m, NHCHCO), 6 47-6 49 (2H, d, J = 8Hz), 6 85 (2H, d, J = 8Hz), 7 03-7 05 (1H, m) 7 16-7 26 (2H, m) and 7 75-7 78 (1H, m) 7 35-7 37 (2H, m) [8H aromatic + 2 NH], MS (m/z) 425 2 (M+l)
Example 16 Preparation of (S)-3-(4-Amino-phenyl)-2-[((S)-l-(2-methoxy phenyl carbamoyl) -pyrrolidine-2-carbonyl)-amino]-propiomc acid methyl ester
Nitro compound (example 10, 610 mg, 1 29 mmol) was reduced via same procedure as
for compound in example 15 to obtain the title compound as oil Yield 550 mg (96%)
IR(vmax,DCM) 3430,2952, 1740, 1659, 1520, 1489, 1459, 1435, 1367, 1250, 1216, 1120, 1026, 835, 751 cm 1 1H NMR (DMSO-d6) δ 1 77-1 80 (3H, m) and 2 00-2 05 (1H, m) [2 X
CHz -CM, 2 77-2 84 (2H, m, ArQi,) 3 23-3 42 (2H, m, NCH2), 3 54 (3H, s, ArOCHs), 3 82 (3H, s, OCH3), 4 32-4 49 (2H, m, NCHCO and NHCHCO), 4 88 (2H, s, NH2), 6 81 (2H, d, J = 8Hz), 6 83 (2H, d, J = 8Hz), 6 88-7 02 (3H, m) 7 19 (1H, bs), 7 91 (lH,d, J= 8Hz), 8 21 (1H, d, J= 8Hz) [8H aromatic + 2 NH], MS (m/z) 441 2 (M+l)
Example 17 Preparation of (S)-3-C4-Aminophenyl)-2-({(S)-l-[('benzo[l,3]dioxo-5ylmethyl)-carbamoyll-pyrrohdine-2-carbonyl}-amino)propionic acid methyl ester
Nitro compound (example 14, 610 mg, 1 6 mmol) was reduced via same procedure as
for compound in example 15 to obtain title compound as sticky mass Yield 720 mg (96%,)
1H NMR (DMSO-d6) δ 1 78-1 87 (4H, m, CH2CH2), 2 77-2 78 (2H, m, ArCH2,), 3 21-3 35 (2H, m, NHCH2), 3 55 (3H, s, OCH3), 4 11-4 33 (4H, m, NHCH2Ar,NCHCO and NHCHCO), 5 94 (2H, s, OCH2O), 6 44 (2H, d, 6Hz), 6 73 (2H, d, J = 7Hz), 6 79-6 84 (4H, m), 8 06 (1H, d, J = 7Hz) [7H aromatic and 2H NH], MS (m/z) 469 2 (M+l)
Example 18 Preparation of (S)-3-(4-Amino-phenyl)-2-{r(S)-l-(brphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino} propionic acid methyl ester
Nitro compound (example 13, 800 mg, 1 5 mmol) was reduced via same procedure as
for compound in example 15 to obtain title compound as oil Yield 700 mg (93%,
hygroscopic)
*H NMR (DMSO-d6) δ 1 68-2 00 (4H, m, CH2CH2), 2 74-2 77 (2H, m, ArCH^), 3 08-3 23 (2H, m, NCH2), 4 20-4 23 (1H, m, NCHCO), 4 29-4 32 (1H, m, NHCHCO), 6 43 (2H, d, J = 7Hz), 6 81 (2H, d, J = 7Hz), 7 15-7 43 (9H, m), 7 71 (2H, d, J = 9Hz) δ 02 (1H, d, J = 8Hz) [13H aromatic + 2NH], MS (m/z) 487 0 (M+l)
Example 19 Preparation of (S)-3-{4-r(Pyndine-4-carbonyl)-amino]-phenyl}-2-r((S)-l-o-tolylcarbamoyl-pyrrolidine-2-carbonyl)-amino] -propionic acid methyl ester
Isomcotinic acid (58 mg, 0 47 mmol), 1-hydroxybenzotnazole (79 mg, 0 52 mmol)
and N- methyl morphohne (142 mg, 141 mmol) were added to a solution of amine (example
15, 200 mg, 0 47 mmol) in dimethylformamide at about 0°C Reaction mixture was stirred
for about 30 minutes and l-(3-dimethylamino propyl)-3-ethyl-carbodimide hydrochloride (99
mg, 0 52 mmol) was added maintaining same temperature Reaction mixture was then stirred
at room temperature for overnight and diluted with ethyl acetate (10 ml), washed with water
(2x5 ml) Organic phase was dried (anhydrous sodium sulphate) and concentrated under
reduced pressure to obtain the crude product Purification by column chromatography (silica
gel, 100 - 200 mesh, 30 70 ethyl acetate hexane) afforded the title compound as
hygroscopic sticky solid Yield 280 mg (90%)
IR(vmax, DCM) 3450, 2923, 2852, 1744, 1654, 1533, 1455, 1364, 1210, 1120, 754 cm \ lH NMR (CDCI3) δ-1 84-2 00 (3H, m) and 2 25-2 29 (1H, m) [2 X CH2 -CHJ, 2 22 (3H, s,
ArCH3), 2 88-3 06 (2H, m, ArCtJb), 3 21-3 45 (2H, m, NCH^), 3 75 (3H, m, OCH3) 4 53-4 56 (1H, m, NCHCO), 4 84-4 86 (1H, m, NHCHCO), 6 97-7 20 (5H, m), 7 37-7 40 (2H, m), 7 61 (2H, d, J = 5Hz), 7 66-7 74 (3H, m) δ 78-8 80 (2H, d, J = 4Hz) [aromatic + 2 NH], MS (m/z) δ30 2(M+1)
Example 20 Preparation of (S)-2-{rS-l-(2-Methoxy-phenylcarbamoyl)-pyrrolidine-2-carbonyll-amino}-3-{4-[(pyndine-4-carbonyl)-aminol-propionic acid methyl ester
Amine (example 16, 150 mg, 0 34 mmol) was reacted with isonicotinic acid (65 mg,
0 34 mmol) via same method as for compound in example 19 to obtain the crude product
Purification by column chromatography (silica gel, 100 - 200 mesh, 3 97, methanol
dichloromethane) afforded the title compound as oil Yield 140mg(75%)
IR (vmax, DCM) 3428,2926,2852, 1743, 1663, 1601, 1534, 1460, 1368, 1251, 1120, 1025, 753 cm \ 1H NMR (DMSO-d6) δ 1 76-1 98 (4H, m) [2 X CH2 -CH2], 2 95-3 01(2H, m, ArCHz), 3 20-3 42 (2H, m, NCH2), 3 57 (3H, s, ArOCH3), 3 82 (3H, m, OCH3), 4 32-4 36 (1H, m, NCHCO), 4 49-4 56 (1H, m, NHCHCO), 6 86-6 98 (3H, m), 7 19-7 23 (2H, m), 7 655 (2H, d, J = 9Hz), 7 83-7 91 (2H, m), 8 32-8 35 (1H, m), 8 78 (2H, d, J = 6Hz) [aromatic], MS (m/z) δ46 1 (M+l)
Example 21 Preparation of (S)-2-({(S)-l-[(Benzo[l,3]dioxol-5-ylmethyl)-carbamoyl]-pyrrolidine-2-carbonyl}-amino)-3{4-[(pyridine-4-carbonyl)-aminol-phenyl}-propionicacid methyl ester
Title compound was synthesized using same procedure as for compound in example
19 using amine (example 17, 150 mg, 0 32 mmol) and isonicotinic acid (39 mg, 0 32mmol)
Purification by column chromatography (silica gel, 100 - 200 mesh, 4 96 methanol
dichloromethane) afforded the title compound as yellow solid Yield 160 mg (87%)
IR (vmax, DCM) 3420, 1739, 1637, 1537, 1443, 1325, 1251, 1038, 753 cm ', 1HNMR (DMSO-d6) δ 1 70-1 97 (4H, m, CH2CH2), 2 94-3 06 (2H, m, ArCHz), 3 19-3 38 (2H, m, NHCH2), 3 61 (3H, s, OCH3), 4 11-4 16 (2H, m, NHCHzAr), 4 24-4 27 (1H, NCHCO), 4 45-4 46 (1H, m, NHCHCO), 5 94 (2H, s, OCH2O), 6 71-6 83 (4H, m), 7 185 (2H, d, J = 8Hz), 7 665 (2H, d, J = 8Hz), 7 84 (2H, d, J = 6Hz), 8 20 (1H, d, J = 7Hz) and 8 77 (2H, d, J = 6 Hz) [11H aromatic and 2HNH], 10 43 (1H, s, NH), MS (m/z) δ74 1 (M+l)
Example 22 Preparation of (S)-2-{[(S)-l-(Biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-3-{4-[(pyndine-4-carbonyl)-amino]-phenyl}-propionic acid methyl ester
Amine (example 18,150 mg, 0 3 mmol) and isonicotinic acid (38 mg, 0 3mmol) were
reacted via same procedure as for compound in example 19 to obtain the crude product
Purification by column chromatography (silica gel, 100 - 200 mesh, 3 97, methanol
dichloromethane afforded the title compound as yellow oil Yield 155 mg (85%)
IR (vmax, DCM) 3489, 2924, 1742, 1659, 1528, 1445, 1412, 1356, 1273, 1213, 753 cm ', 1H
NMR (DMSO-d6) δ 1 73-1 98 (4H, m, CH2CH2), 2 94-3 22 (4H, m, ArCH2 and NCH2), 3 58
(3H, s, OCH3), 4 20-4 23 (1H, m, NCHCO), 4 43-4 45 (1H, m, NHCHCO), 7 16-7 43 (10H, m), 7 64-7 72 (3H, m), 7 84 (2H, d, J = 6Hz), 8 18 (1H, d, J= 7Hz), 8 78 (2H, d, J = 6Hz) [17H aromatic and NH], MS (m/z) δ92 2 (M+l)
Example 23 Preparation of (S)-3-(4-Benzoylamino-phenyl)-2-{[(S)-(biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-propionic acid methyl ester
Tnethyl amine (31 mg, 0 3 mmol) and benzoyl chloride (43 mg, 0 3 mmol) were
added to cold solution of amine (example 18, 150 mg, 0 3 mmol) in dichloromethane (10
ml) Reaction mixture was stirred at same temperature for about lhr and diluted with
dichloromethane (10 ml), which was then washed with water, aqueous sodium bicarbonate
and brine (10 ml each) Organic phase was dried (anhydrous sodium sulphate) and
concentrated in vacuum to obtain the crude product Purification by column chromatography
(silica gel, 100 - 200 mesh, 3 97 methanol dichloromethane) afforded the title compound as
yellow oil Yield 133mg(73%)
IR (vmax, DCM) 3425, 2924, 1744, 1651, 1521, 1448, 1412, 1360, 1266, 1214, 1121, 755, 705 cm ', 1H NMR (DMSO-d6) δ 1 73-1 77 (3H, m) and 1 81-1 83 (1H, m) [CH2CH2], 2 91-3 34 (4H, m, NCH2 and ArCH2), 3 58 (3H, s, OCH3), 4 21-4 24 (1H, m, NCHCO), 4 42-4 44 (1H, m, NHCHCO), 7 15-718 (3H, m), 7 25-7 73 (14H, m) and 7 93 (2H, d, J= 9H) δ 17 (1H, d, J = 8Hz) [18H aromatic and 2 NH], MS (m/z) 613 2 (M+l)
Example 24 Preparation of 4-[4((S)-2-{[(S)-l-(Biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyll-amino}-2-methoxycarbonyl-ethyl-)-phenylcarbamoyll-piperidine-l-carboxyhc acid tert-butyl ester
Title compound was synthesized via same procedure as for compound in example 19
using amine (example 18, 300 mg, 0 6 mmol) and Boc-isonipecotic acid (141 mg, 0 6mmol)
Purification by column chromatography (silica gel, 100 - 200 mesh, 3 97, methanol
dichloromethane) afforded the title compound as off white solid Yield 318 mg (74%)
IR U DCM) 3424, 1741, 1654, 1521, 1448, 1365, 1167, 736 cm 1 1H NMR (DMSO-d6)
δ 1 40 (9H, s, C(CH3)3), 1 47-1 76 (8H, m, 4 X CH2), 2 75-2 95 (5H, m), 3 11-3 21 (3H, m) [ArCHs and 3 X NCHJ, 3 58 (3H, s, OCH3), 4 19-4 21 (1H, m, NCHCO), 4 41-4 42 (1H, m, NHCHCO), 7 11 (2H, d, J = 8 Hz), 7 21-7 48 (9H, m), 7 66 (1H, d, J = 8Hz), 8 13 (1H, d, J = 8Hz) [13H aromatic + NH], MS (m/z) 698 3 (M+l)
Example 25 Preparation of (S)-3-{4-[(Pyridine-4-carbonyl)-amino]-phenyl}-2-[((S)-l-o-tolylcarbamoyl-pyrrohdine-2-carbonyl)-amino] -propionic acid (Compound No 7)
Lithium hydroxide (16 mg, 0 37 mmol) was added to a solution of methyl ester
(example 19, 200mg, 0 37 mmol) in tetrahydrofuran (3 ml), methanol (1 ml) and water (1
ml) Reaction mixture was stirred for a bout 3 hr and concentrated to obtain the crude product,
which was taken in water (10 ml) and extracted with ethyl acetate (2x10 ml) Aqueous layer was acidified with sodium hydrogen sulphate and extracted with ethyl acetate (2x10 ml) This combined organic phase was washed with water and brine (10 ml each), dried (anhydrous sodium sulphate) and concentrated to obtain the title compound as sticky mass Yield 120 mg (62%) (mixture of diastereomers, 86 12 by HPLC, tR = 13 9 &14 5 mm , Channel 486)
1H NMR (DMSO-d6) δ 1 84-1 98 (4H, m, 2 X CH2 -CH2), 2 15 (3H, s, ArCH,), 2 93-3 02 (2H, m, ArCH2), 3 44-3 53 (2H, m, NCH2), 4 34-4 43 (2H, m, NCHCO and NHCHCO), 6 99-7 33 (5H, m), 7 59-7 65 (3H, m), 7 83-7 85 (2H, m), 7 99 (1H, d, J = 7Hz) and 8 775 (2H, d, J = 5Hz) [aromatic] MS (m/z) δ16 2(M+1)
Example 26 Preparation of (S)-2-{[S-l-(2-Methoxy-phenylcarbamoyl)-pyrrolidine-2-carbonyl]-amino}-3-{4-[(pyridine-4-carbonyl)-aminophenyl]-propiomc acid (Compound No 8)
Ester (example 20,140 mg, 0 25 mmol) was hydrolyzed via same procedure as for
compound in example 25 to obtain the title compound as light yellow solid Yield 115 mg
(85%)
IR(vmax,DCM) 3431, 3309,2951, 1748, 1670, 1602, 1536, 1460, 1434, 1414, 1366, 1248, 1165, 1121, 1027, 955, 752 cm 1HNMR (DMSO-d6) δ 1 76-2 05 (4H, m, CH2-CH2), 2 95-2 98 (1H, m) and 3 05-3 07 (1H, m) [Ar CH2] 3 38-3 41 (1H, m) and 3 50-3 52 (1H, m) [NCH2], 3 81 (3H, s, OCH3), 4 33-4 36 (1H, m, NCHCO), 4 43-4 47 (1H, NHCHCO), 6 87-6 90 (1H, m), 6 98-6 99 (2H, m), 7 22-7 25 (2H, m), 7 59-7 62 (2H, m),7 78-7 84 (3H, m) and 8 75-8 77 (2H, m) [aromatic], MS (m/z) δ32 2 (M+l)
Example 27 Preparation of (S)-2-({(S)-l-[(Benzon,31dioxol-5-ylmethyl)-carbamoyl]-pyrrohdine-2-carbonyl}-amino)-3{4-[(pyndine-4-carbonyl)-amino]-phenyl}-propionicacid (Compound No 9)
Methyl ester (example 21, 150 mg, 0 26 mmol) was hydrolyzed via same procedure
as for compound in example 25 to obtain desired compound as cream-colored solid Yield
130mg(89%)
lH NMR (DMSO-d6) δ 1 84-1 98 (4H, m, CH2CH2), 2 88-3 08 (2H, m, ArCHz), 3 20-3 34 (2H, m, NHCHj.), 4 11-4 27 (4H, m, NHCHaAr, NHCHCO and NCHCO), 5 93 (2H, s, OCH2O), 6 71-6 83 (4H, m), 7 185 (2H, d, J = 8Hz), 7 65 (2H, d, J = 8Hz), 7 84 (2H, d, J = 6Hz), 7 99 (1H, d, J = 7Hz) and 8 77 (2H, d, J = 6 Hz) [11H aromatic and 2H NH], 10 43 (lH,s,NH) MS (m/z) δ60 0 (M+l)
Example 28 Preparation of (S)-2-{[(S)-l-(Biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbony 1]-amino) -3- {4-[(pyndine-4-carbonyl)-amino]-phenyl) -propionic acid (Compound No 10)
Methyl ester (example 22, 150 mg, 0 25 mmol) was hydrolyzed via same procedure as for compound in example 25 to obtain title compound as cream-colored solid Yield 130 mg (89%)
1H NMR (DMSO-d6) δ 1 72-1 82 (4H, m, CH2CH2), 2 91-3 35 (4H, m, ArCH2 and NCH2), 4 20-4 22 (1H, m, NCHCO), 4 36-4 48 (1H, m, NHCHCO), 7 17-7 43 (10H, m), 7 64-7 70 (3H, m), 7 86 (2H, d, J = 6Hz), 7 91 (1H, d, J= 7Hz), 8 78 (2H, d, J = 6Hz)[17H aromatic and NH], MS (m/z) δ78 2 (M+l)
Example 29 Preparation of (S)-3-(4-Benzoylamino-phenyl)-2-{ri-(S)-(biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-propionic acid (Compound No 11)
Methyl ester (example 23, 150 mg, 0 25 mmol) was hydrolyzed via same procedure
as for compound in example 25 using L1OHH2O (10 mg, 0 25 mmol) to obtain title
compound as cream-colored solid Yield 140 mg (96%)
IR (vmax, DCM) 3424, 1736, 1654, 1525, 1449, 1412, 1364, 1322, 1266, 1118, 754, 706 cm1, 1H NMR (DMSO-d6) δ1 72-1 83 (4H, m, CH2CH2), 2 93-3 19 (4H, m, NCH2 and ArCH2), 4 20-4 23 (1H, m, NCHCO), 4 43-4 45 (1H, m, NHCHCO), 7 15-7 68 (17H, m) and 7 92-7 94 (3H, m) [18H aromatic and 2 NH], MS (m/z) δ77 1 (M+l)
Example 30 Preparation of 4-[4((S)-2-U(S)-l-(Biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-2-carboxy-ethyl)-phenylcarbamoyl]-piperidine-l-carboxyhc acid tert-butyl ester (Compound No 12)
Methyl ester (example 24, 300 mg, 0 43 mmol) was hydrolyzed via same procedure
as for compound in example 25 using L1OHH2O (18 mg, 0 43 mmol) to obtain title
compound as off white solid Yield 270mg(92%)
1H NMR (DMSO-d6) δ1 45 (9H, s, C(CH3)3), 1 49-1 99 (8H, m, 4 X CH2), 2 77-3 19 (8H, m, ArCH2 and 3 X NCH2), 4 22-4 23 (1H, m, NCHCO), 4 42-4 45 (1H, m, NHCHCO), 7 09-7 48 (6H, m), 7 69 (1H, d, J = 8Hz) and 7 87 (1H, d, J = 7Hz ) [aromatic], MS (m/z) 684 4 (M+l)
Example 31 Preparation of (S)-2-{[(S)-l-(Biphenyl-2-ylcarbamoyl)-pyrrolidine-2-carbonyll-amino}-3-{4-|"(pipendine-4-carbonyl)-amino]-phenyl}-propionic acid TFA salt (Compound No 13)
Boc amine (example 30, 100 mg, 0 14 mmol) was stirred with 30% trifluoroacetic
acid- dichloromethane (5 ml) for about 3h and concentrated under high vacuum to obtain the
desired compound as sticky mass Yield 95 mg (95%)
1H NMR (DMSO-d6) δ 1 76-1 95 (8H, m, 4 X CH2], 2 87-2 97 (5H, m) and 3 10-3 21 (1H, m) and 3 32-3 70 (2H, m) [ArCfcb, NHCOCH, 3 X NCH2), 4 19-4 22 (1H, m, NCHCO), 4 35-4 37 (1H, m, NHCHCO), 7 11-7 48 (12 H, m), 7 68 (1H, d, J = 8Hz), 7 945 (1H, d, J = 8Hz), 8 31 (1H, bs), 8 55 (1H, bs) [13H aromatic and 3HNH], MS (m/z) δ84 2 (M+l)
Example 32 Preparation of (S)-2-{r(S)-l-(Biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-3-(4-nitro-phenyl)-propionic acid (Compound No 14)
LiOH H20 (24 mg, 0 57 mmol) was added to a solution of methyl ester (example 13,
300mg, 0 57 mmol) in tetrahydrofuran (3 ml), methanol (1 ml) and water (1 ml) Reaction
mixture was stirred for about 3 hr and concentrated to obtain the crude oil, which was taken
in water (10 ml) and extracted with ethyl acetate (2 x 10 ml) Aqueous layer was acidified
with sodium hydrogen sulphate and extracted with ethyl acetate (2x10 ml) This combined
organic phase was washed with water and brine (10 ml each), dried over anhydrous sodium
sulphate and concentrated to obtain the desired compound as sticky mass Yield 240 mg
(82%)
1H NMR (DMSO-d6) δ 1 70-2 07 (4H, m, CH2CH2), 3 00-3 23 (4H, m, NCH2 and ArCHa), 4 17-4 19 (1H, m, NCHCO), 4 45-4 47 (1H, m, NHCHCO), 7 18-7 50 (10H, m), 7 65-7 67 (1H, m) and 8 06-8 09 (2H, m) [aromatic], MS (m/z) δ03 2 (M+l)
Example 33 Preparation of (S)-3-(4-Amino-phenyl)-2-{[(S)-l-(biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbony 11-amino} propionic acid (Compound No 15)
10% Palladium on carbon (50 mg, dry) was added to a solution of mtro compound
(example 32, 50 mg, 0 29 mmol) in methanol (10 ml) Reaction mixture was shook under
hydrogen atmosphere at 50 psi for aboutl hr and filtered through cehte pad Filtrate was
concentrated to obtain the title compound as sticky mass Yield 120 mg (85 %) (mixture of
diastereomers, 87 3 by HPLC, tR = 14 6 &15 5 mm, Channel 486)
1H NMR (DMSO-d6) δ 1 70-2 07 (4H, m, CH2CH2), 2 60-2 33 (4H, m, NCH2 and ArCIi), 4 23-4 45 (2H, m, NCHCO and NHCHCO), 6 48 (2H, d, J = 7Hz), 6 86 (2H, d, J = 7Hz), 718-7 44 (9H, m), 7 68-7 82 (2H, m) [13H aromatic + 2NH], MS (m/z) 473 2 (M+l)
Example 34 4-{4-f(S)-2-({(S)-l-[(Benzo[l,3]dioxol-5-ylmethyl)carbamoyl]-pyrrohdine-2-carbonyl} -amino)-2-carboxy-ethyl] -phenylcarbamoyl} -pipendine-1 -carboxylic acid tert-butyl ester (Compound No 16)
This compound was prepared following the same procedure as for compound in
example 30 starting from the compound of example-17
1H NMR (DMSO-d6) δ1 40 (9H, s, C(CH3)3), 1 44-1 90 (8H, m,4 X CH2), 2 53-2 54 (1H, m, NHCOCH), 2 79-3 01 (4H ,m NCH2 and ArCHa), 3 18-3 20-3 34 (2H, m), 3 97-4 02 (2H, m) [2 X NHCH2], 4 13 (2H, s, ArCEbNH), 4 24-4 45 (2H, m, NCHCO and NHCHCO), 5 91 (2H, s, OCH2O), 6 73-6 81 (3H, m), 7 10-7 13 (2H, m), 7 42-7 45 (2H, m) [aromatic], MS (m/z) 666 3 (M+l)
Example 35 Preparation of l-(Biphenyl-2-ylcarbamoyl)-pyrrolidine-2-carboxyhc acid methyl ester
Ethyldnsopropylamine (9 76g, 75 5 mmol) was added to a solution of proline methyl ester hydrochloride (5g, 30 2mmol) in dry tetrahydrofuran (100 ml) at about 0°C A solution of carbamate (example 11, lOg, 30 2 mmol) in tetrahydrofuran (100 ml) was added drop wise to cold reaction mixture Reaction mixture was stirred at room temperature for overnight, concentrated in vacuum to 1/3 of its volume and diluted with ethyl acetate (100 ml) Organic phase was washed with water and brine (2 X 50 ml each), dried (anhydrous sodium sulphate) and concentrated in vacuum to obtain the crude compound Purification by column chromatography (silica gel, 100 -200 mesh 70 30 ethyl acetate hexane) afforded title compound Yield 8 7g (89%)
IR (vmax, DCM) 3429, 2953, 1745, 1719, 1673, 1584, 1524, 1448, 1356, 1281, 1176, 1009, 753, 705 cm', 1H NMR (DMSO-d6) δ 1 78-1 89 (3H, m), 2 08-2 14 (1H, m) [CH2CH2, 3 21-3 22 (2H, m, NCH2), 3 59 (3H, s, OCH3), 4 25-4 26 (1H, m, NCHCO), 7 20-7 57 (10 H, m, 9H aromatic + 1NH), MS (m/z) 325 0 (M+l)
Example 36 Preparation of l-(Biphenyl-2-yIcarbamoyl)-pyrrolidine-2-carboxyhc acid
Methyl ester (example 35, 8 23g, 25 4 mmol) was hydrolyzed via same procedure as for compound in example 32 to obtain the title compound as white solid Yield 6 25g (80%)
Example 37 Preparation of (S)-2-{[-l-(Biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-3-(4-nitro-phenyl)-propiomc acid methyl ester
Title compound was prepared via same procedure as for compound in example 2
using acid (example 36, 6 4 mmol) and amine (example 1, 6g, 6 45 mmol) Purification of
crude compound by column chromatography (silica gel, 100-200 mesh, 4 96 methanol
Dichloromethane) afforded pure compound Yield 2 8g (84%)
IR (vmax, DCM) 3426, 3057, 2953, 1744, 1674, 1521, 1446, 1348, 1272, 1215, 1112, 858, 737, 703 cm1, 1H NMR (DMSO-d6) δ 1 68-1 90 (4H, m, CH2CH2), 3 02-3 21 (4H, m, NCH2 and ArCH2), 3 60 (s) and 3 63 (s) (3H, OCH,), 4 16-4 19 (1H, m, NCHCO), 4 52-4 57 (1H, m, NHCHCO), 7 19-7 50 (10H, m), 7 68 (1H, d, J = 9Hz) and 8 08 (2H, d, J = 9Hz) [aromatic], 8 30 (1H, d, J = 7Hz, NH), MS (m/z) δ17 3 (M+l)
Example 38 Preparation of (S)-3-(4-Amino-phenyl)-2-{[l-(biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino} propionic acid methyl ester
Nitro compound (example 37, 1 7g, 3 3mmol) was reduced using same procedure as
for compound in example 3 using 10% palladium on carbon (500 mg, dry) in methanol (30
ml) to obtain the title compound as sticky mass Yield 1 5g(94%)
Example 39 Preparation of (S)-3-(4-Acetylamino-phenyl)-2-{[l-(biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-propionic acid methyl ester
Acetic anhydride (41 mg, 0 4 mmol) and tnethylamine (42 mg, 0 4 mmol) were
added to a solution of amine (example 38,200 mg, 0 4 mmol) in dichloromethane at about
0°C Reaction mixture was stirred at same temperature for about lh and at room temperature
for overnight Reaction mixture was diluted with dichloromethane (10ml) and washed with
water and brine (20 ml each) Organic layer was dried (anhydrous sodium sulphate) and
concentrated to obtain the crude compound Purification by column chromatography (silica
gel, 100-200 mesh, 3 97 methanol dichloromethane) afforded the title compound as oil
Yield 170 mg (78 3%)
IR (vmax, DCM) 3427, 1742, 1655, 1518, 1445, 1410, 1367, 1210, 753, 698 cm ', 1H NMR (DMSO-d6) δ 1 66-1 89 (4H, m, CH2CH2), 2 01 (3H, s, COCH3), 2 87-2 94 (2H, m, ArCH2),
3 07-3 09 (1H, m) and 3 21-3 24 (1H, m) [NCHJ, 3 56 (s) and 3 59 (s) [3H, OCH3], 4 20-
4 22 (1H, m, NCHCO), 4 39- 4 41 (1H, m, NHCHCOOH), 7 08-7 46 (10H, m), 7 71 (1H, d, J = 8Hz), 8 13 (1H, d, J = 8Hz) [aromatic], MS (m/z) δ29 1 (M+l)
Example 40 Preparation of (S)-2-{[l-(Biphenyl-2-ylcarbamoyl)-pyrrohdme-2-carbonyl]-amino}-3-[4-(2,2,2-tnfluoro-acetylamino)-phenyl]-propionic acid methyl ester
Title compound was prepared via same procedure as for compound in example 39
using amine (example 38, 200 mg, 0 4 mmol) and trifluoroacetic anhydride (87 mg, 0 4
mmol) in presence of tnethylamine (42 mg, 0 4 mmol) Purification by column
chromatography (silica gel, 100-200 mesh, 3 97 methanol dichloromethane) afforded the
title compound as oil Yield 140 mg (58 5%)
1H NMR (DMSO-d6) δ 1 66-1 92 (4H, m, CH2CH2), 2 90-3 20 (4H, m, ArCH2, NCH2), 3 57 (s) and 3 61 (s) [3H, OCH3], 4 16-4 19 (1H, m, NCHCO), 4 45- 4 47 (1H, m, NHCHCOOMe), 7 17-7 55 (13H, m) 7 71 (1H, d, J = 8Hz), 8 12-8 14 (1H, m) [13H aromatic + 2NH],MS(m/z) δ83 2 (M+l)
Example 41 Preparation of (S)-2- {[ 1 -(Biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl] -amino }-3-(4-methanesulfonyl amino-phenyD-propionic acid methyl ester
Methanesulfonyl chloride (47 mg, 0 41 mmol) and tnethylamine (45 mg, 0 45 mmol)
were added to a solution of amine (example 38, 200 mg, 0 41 mmol) in dichloromethane (10
ml) at about 0°C Reaction mixture was stirred for overnight at room temperature and diluted
with dichloromethane (10 ml) Organic layer was washed with water and brine (2X 10ml
each) Organic layer was dried (anhydrous sodium sulphate) and concentrated to obtain the
crude compound Purification by column chromatography (silica gel, 100-200 mesh, 3 97, methanol dichloromethane) afforded the title compound as oil Yield 85 mg (37%)
IR(vmax, DCM) 3426, 2926, 1743, 1658, 1584, 1517, 1447, 1338, 1219, 1152, 972, 755, 705 cm \ 1H NMR (DMSO-d6) δ 1 56-1 87 (4H, m, CH2CH2), 2 84 (3H, s, S02CH3), 2 85-3 23 (4H, m, NCH2 and ArCIfc), 4 17-4 21 (1H, m, NCHCO), 4 43-4 44 (1H, m, NHCHCO), 7 06-7 44 (13H, m), 7 69-7 71 (1H, m), 8 11-8 14 (1H, m) [13H aromatic and 2H NH], MS (m/z) δ65 0(M+1)
Example 42 Preparation of (S)-3-(4-Benzenesulfonylamino-phenyl)-2-{[l-(biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-propionic acid methyl ester
Title compound was prepared via same procedure as for compound in example 41
using amine (example 38, 200 mg, 0 41 mmol) and benzenesulfonyl chloride (73 mg, 0 41
mmol) Purification by column chromatography (silica gel, 100-200 mesh, 4 96, methanol
dichloromethane) afforded the title compound as white solid Yield 180 mg (70%)
IR(vmax, DCM) 3426, 2951, 2870, 1743, 1662, 1584, 1516, 1447, 1346, 1221, 1161, 1092, 923, 755, 689, 583 cm 1HNMR (CDCI3) δ 1 78-2 22 (4H, m, CH2CH2), 2 85-3 06 (4H, m, NCH2 and ArCH2), 3 68 (3H, bs, OCH3), 4 43-4 45 (1H, m, NCHCO), 4 72-4 76 (1H, m, NHCHCO), 6 43-6 46 (1H, m), 6 75-6 76 (2H, m), 6 95-6 99 (2H, m), 7 24-7 50 (14H, m), 7 62-7 65 (2H, m), 8 16-8 19 (1H, m) [18 H aromatic and 2 H NH), MS (m/z) 627 1 (M+l)
Example 43 Preparation of (S)-2-{[l-(Biphenyl-2-ylcarbamoyl)-pyrrolidine-2-carbonylj-amino}-3-(4-toluene-4-sulfonyl amino)phenyl)-propionic acid methyl ester
Title compound was prepared via same procedure as for compound m example 41
using amine (example 38, 200 mg, 0 41 mmol) and 4-toluenesulfonyl chloride (78 mg, 0 41
mmol) Purification by column chromatography (silica gel, 100-200 mesh, 3 97, methanol
dichloromethane) afforded the title compound as sticky mass Yield 230 mg (80%)
IR(vmax,DCM) 3426,2925, 1744, 1665, 1584, 1516, 1447, 1343, 1219, 1159, 1092,923, 815, 755, 706cm ', lH NMR (DMSO-d6) δ 151-19 (4H, m, CH2CH2), 2 29 (3H, s, ArCH2), 2 82-3 22 (4H, m, NCH2 and ArCH2), 3 48 (s) and 3 52 (s) [3H, OCH3], 4 11-4 13 (1H, m, NCHCO), 4 32-4 33 (1H, m, NHCHCO), 6 91-7 01 (4H, m), 7 15-7 42 (11H, m), 7 57-7 66 (3H, m), 8 29 (1H, bs) [17 H aromatic and 2 H NH), MS (m/z) 641 1 (M+l)
Example 44 Preparation of (S)-3-(4-Acetylamino-phenyl)-2-{[l-(biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-propiomc acid (Compound No 2)
Ester (example 39, 170 mg, 0 32 mmol) was hydrolyzed via same procedure as for
compound in example 32 using LiOH H2O (14 mg, 0 35 mmol) to obtain the title compound
as white solid Yield 140mg(85%)
1HNMR (DMS0-d6) δ1 71-1 91 (4H, m, CHzCHz), 2 01 (3H, s, COCH3), 2 87-3 18 (4H, m, ArCHz, NCH2), 4 19-4 22 (1H, m, NCHCO), 4 41- 4 42 (1H, m, NHCHCOOH), 7 08-7 44 (13H, m), 7 69-7 72 (1H, m) [aromatic], 7 88-7 89 (1H, m, NH), MS (m/z) δ15 2 (M+l)
Example 45 Preparation of (S)-2-{[l-Biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amimo}-3-[4-(2,2,2-tnfluoro-acetylammo)-phenyll-propionic acid (Compound No 3)
Ester (example 40, 140 mg, 0 24 mmol) was hydrolyzed via same procedure as for
compound in example 32 using LiOH H2O (15 mg, 0 24 mmol) to obtain the title compound
as white solid Yield 124mg(91%)
1H NMR (DMSO-d6) δ 1 71-1 92 (4H, m, CH2CH2), 2 90-3 21 (4H, m, ArCEb, NCH2), 4 19-4 22 (1H, m, NCHCO), 4 42- 4 43 (1H, m, NHCHCOOH), 7 17-7 52 (12H, m) and 7 51-7 52 (1H, m) [aromatic], MS (m/z) δ69 0 (M+l)
Example 46 (S)-2-{ri-(Biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyll-amino}-3-(4-methanesulfonyl amino-phenyl)-propionic acid (Compound No 4)
Ester (example 41, 75 mg, 0 13 mmol) was hydrolyzed via same procedure as for
compound in example 32 using LiOH H2O (6 mg, 0 15 mmol) to obtain the title compound
as white colored solid Yield 70 mg (98%) M Pt = 125°C
1HNMR (DMSO-d6) δ 1 57-1 85 (4H, m, CHzCHa), 2 84 (3H, s, S02CH3), 2 85-3 19 (4H, m, NCH2 and ArCH2), 4 17-4 21 (1H, m, NCHCO), 4 39- 4 40 (1H, m, NHCHCO), 7 05-7 44 (13H, m), 7 69-7 72 (1H, m) [13H aromatic and 1H NH], MS (m/z) δ51 0 (M+l)
Example 47 Preparation of (S)-3-(4-Benzenesulfonylamino-phenyl)-2-{[l-(biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-propionic acid (Compound No 5)
Ester (example 42, 160 mg, 0 26 mmol) was hydrolyzed via same procedure as for
compound in example 32 using LiOH H2O (12 mg, 0 28 mmol) to obtain the title compound
as light yellow solid Yield 160 mg (100%) M Pt = 130°C
1HNMR (DMSO-d6) δ 1 62-1 91 (4H, m, CH2CH2), 2 79-3 14 (4H, m, NCHzand ArCH^), 4 14-4 17 (1H, m, NCHCO), 4 33-4 34 (1H, m, NHCHCO), 6 93-7 05 (4H, m), 7 25-7 56 (13H, m), 7 70-7 73 (3H, m), [18 H aromatic and 2 H NH), MS (m/z) 613 1 (M+l)
Example 48 Preparation of (S)-2-{[l-(Biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-3-(4-toluene-4-sulfonyl amino)phenyl)-propiomc acid (Compound No 6)
Ester (example 43, 200 mg, 0 31 mmol) was hydrolyzed via same procedure as for
compound in example 32 using LiOH H2O (15 mg, 0 35 mmol) to obtain the title compound
as white solid Yield 160mg(83%) MPt = 132°C
IR(vmax,DCM) 3424,3255,2926, 1733, 1651, 1584, 1516, 1448, 1366, 1339, 1225, 1158, 1092, 923, 815, 756 cm ', 1H NMR (DMSO-d6) δ 1 49-1 9 (4H, m, CH2CH2), 2 31 (3H, s, ArCHs), 2 75-3 14 (4H, m, NCH2 and ArCHz), 4 13-4 16 (1H, m, NCHCO), 4 33-4 34 (1H,
m, NHCHCO), 6 93-7 03 (4H, m), 7 17-7 44 (11H, m), 7 58-7 61 (2H, m), 7 71 (1H, bs) 7 88 (1H, bs) [17 H aromatic and 2 H NH), MS (m/z) 627 1 (M+l)
Example 49 Preparation of (S)-2-{ri-(Biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-3-(4-fluoro-phenyl)-propiomc acid methyl ester
Title compound was prepared via same procedure as for ester in example 2 using acid
(example 36, 200 mg, 0 66 mmol) and (S)- 4-fluorophenyl alanine methyl ester
hydrochloride (142 mg 0 66mmol) Purification by column chromatography (silica gel, 100-
200 mesh, 2 98, methanol dichloromethane) afforded the title compound as sticky mass
Yield 280mg(87%)
1H NMR (DMSO-d6) δ 1 69-1 90 (4H, m, CH2CH2), 2 89-3 26 (4H, m, NCH2 and ArCHz), 3 59 (s) and 3 61 (s) [3H, OCH3], 4 18-4 19 (1H, m, NCHCO), 4 43-4 48 (1H, m, NHCHCO), 7 00-7 03 (2H, m), 7 18-7 44 (11H, m), 7 68-7 69 (1H, m) [13H aromatic and 1HNH], 8 16 (1H, bs, NH), MS (m/z) 490 1 (M+l)
Example 50 Preparation of (S)-2-{[l-(Biphenyl-2-ylcarbamoyl)-pyrrolidine-2-carbonyl]-amino}-3-(4-hydroxy-phenyl)-propionic acid ethyl ester
Title compound was prepared via same procedure as for ester in example 2 using acid
(example 36, 1 5g, 4 84 mmol) and tyrosine ethyl ester hydrochloride (1 19g, 4 84 mmol)
Purification by column chromatography (silica gel, 100-200 mesh, 3 97 methanol
dichloromethane) afforded the title compound as sticky mass Yield 2 2g (90 9%)
1H NMR (DMSO-d6) δ 1 06-1 14 (3H, m, CH2CH3), 1 72-1 95 (4H, m, CH2CH2), 2 75-
3 21(4H, m, NCH2 and ArCH2), 3 99-4 04 (2H, m, CH2CH3), 4 19-4 21 (1H, m, NCHCO),
4 31-4 32 (1H, m, NHCHCO), 6 59-6 64 (2H, m), 6 94-6 98 (2H, m), 7 17-7 44 (8H, m),
7 70-7 71 (1H, m), 8 05 (1H, bs) [13 H aromatic + NH], 9 21 (1H, bs, NH), MS (m/z) δ02 1 (M+l)
Example 51 Preparation of (S)-2-{[l-(Biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyll-amino}-3-[4-(2-morpholin-4-yl-ethoxy)-phenyl}-propionic acid ethyl ester
4-(2-Chloroethyl) morphohne hydrochloride (93 mg, 0 49 mmol) and potassium
carbonate (137 mg, 0 98 mmol) were added to a solution of compound of example 50 (250
mg, 0 49 mmol) Reaction mixture was refluxed for about 18 h and filtered Filtrate was
concentrated in vacuum to obtain the crude compound Purification by column
chromatography (silica gel, 100-200 mesh, 3 97 methanol dichloromethane) afforded the
title compound as sticky mass Yield 210mg(69%)
1 H NMR (DMSO-d6) δ 1 07-1 15 (3H, m, CH2CH3), 1 74-1 91 (4H, m, CH2CH2), 2 42-2 49 (4H, m, N(CH2)2), 2 62-2 65 (2H, m, NCH2), 2 89-3 32 (4H, m, NCH2 and ArCHj,), 3 54-3 57 (4H, bs, OCH2), 3 97-4 06 (4H, m, ArOCHj, and OCH2), 4 24-4 47 (2H, m, NHCHCO and
NCHCO), 6 73-6 80 (2H, m), 7 05-7 10 (2H, d, J = 6Hz), 7 16-7 44 (9H, m), 7 71 (1H, bs), 8 08-8 31 (1H, m) [13H aromatic + 2 NH), 7 46-7 63 (1H, m, NH), MS (m/z) 615 2 (M+l)
Example 52 Preparation of (S)-2-{ri-(Biphenyl-2-ylcarbamoyl)-pyrrolidine-2-carbonyll-amino}-3-(4-fluoro-phenyl)propionic acid (Compound No 17)
Ester (example 49, 280 mg, 1 5 mmol) was hydrolyzed via same procedure as for
compound in example 32 using LiOH H2O (70mg, 1 65 mol) to obtain the title compound
Yield 250mg(92%)
IR(vmax, DCM) 3427, 3272, 3073, 2967, 1722, 1686, 1647, 1563, 1513, 1478, 1447, 1379, 1277, 1234, 1163, 814, 750 cm ', 1H NMR (DMSO-d6) δ 1 58-1 87 (4H, m, CH2CH2), 2 85-3 19 (4H, m, NCH2 and ArCH2,), 4 16-4 21 (1H, m, NCHCO), 4 38-4 43 (1H, m, NHCHCO),
6 97-7 02 (2H, m), 7 18-7 44 (11H, m), 7 66-7 69 (1H, m) [13H aromatic and 1H NH], 7 94-
7 93 (1H, m, NH), MS (m/z) 476 0 (M+l)
Example 53 Preparation of (S)-2-{[l-(Biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-3-(4-hydroxy-phenyl)-propionic acid (Compound No 18)
Ester (example 50, 750mg, 1 5 mmol) was hydrolyzed via same procedure as for
compound in example 32 using LiOH H2O (70mg, 1 65 mol) to obtain the title compound
Yield 500mg(87%)
1H NMR (DMSO-d6) δ 1 63-1 95 (4H, m, CH2CH2,), 2 55-2 83 (1H, m), 2 91-2 95 (1H, m) and 3 08-3 19 (2H, m) [NCH2 and ArCH2,], 4 19-4 20 (1H, m, NCHCO), 4 34-4 38 (1H, m, NHCHCO and), 6 61-6 67 (2H, m), 6 98-7 01 (2H, m), 7 24-7 30 (2H, m)7 36 -7 46 (6H, m), 7 63-7 68 (1H, m) [13 H aromatic], 9 18 (1H, bs, NH), MS (m/z) 474 3 (M+l)
Example 54 Preparation of (S)-2-{[l-(Biphenyl-2-ylcarbamoyl)-pyrrohdine-2-carbonyl]-amino}-3-[4-(2-morphohn-4-yl-ethoxy)-phenyl}-propionic acid (Compound No 19)
Ester (example 51, 187 mg, 0 3 mmol) was hydrolyzed via same procedure as for
compound in example 32 using LiOH H2O (12 mg, 0 3 mmol) to obtain the title compound
as cream colored solid Yield 162mg(89%)
1H NMR (DMSO-d6) δ 1 74-1 91 (4H, m, CH2CH2), 2 42-2 49 (4H, m, N(CH2)2), 2 59-2 65 (2H, m, NCH2), 2 89-3 32 (4H, m, NCHj-and ArCHj), 3 56 (4H, bs, OQh), 3 65-4 12 (4H, m, ArOCH2, NHCHCO and NCHCO), 6 58-6 69 (2H, m), 6 96(2H, d, J = 6Hz), 7 16-7 44 (9H, m) [aromatic], 7 46-7 63 (1H, m, NH), MS (m/z) δ87 2 (M+l)
Example 55 Primary Screen Cell Adhesion Assay
VCAM-1 (l00ng/well) was coated in Maxisorp microtitre module at 4°C overnight
Non-specific blocking was carried out with 3% BSA for 1 hr and the wells washed with TBS
(50mM Tns, 0 15M NaCl pH 7 4, ImM CaCl2, ImM MgCl2, ImM MnCl2) Jurkat J6 cells
were suspended in fresh medium and incubated at 37°C for 2 hrs before the assay Cells were
then washed in TBS solution and 180 µl of cell suspension (5x10 cells/ml in TBS buffer) added per well in VCAM-1 coated wells 20 µl of sample in 50% DMSO and 50% TBS was then added and the cells incubated at 37°C for 1 hr 3 to 5 dilutions of each sample were tested in duplicate After incubation, the non-adherent cells were removed by washing with TBS and the number of adhered cells was quantified by LDH activity estimation The percent adhesion was calculated as compared to control Compounds provided herein showed activities in the range of 1 l->30 µM

WE CLAIM:
1. A compound having the structure of Formula I
(Formula Removed)
and its pharmaceutically acceptable salts, pharmaceutically acceptable solvates,
stereoisomers, tautomers, racemates, regioisomers, prodrugs, metabolites,
polymorphs or N-oxides wherein
ring A including X is a five membered ring wherein X is nitrogen;
Y is-CONH-;
R1 is aryl (optionally substituted with alkyl, aryl or alkoxy) or aralkyl (wherein
aryl group is fused with a cycloalkyl or heterocyclyl group),
R.2 is nitro, amino, hydroxy, halogen, -NHCOR3 (wherein R3 is optionally
substituted heteroaryl, aryl, alkyl or heterocyclyl, -NHSO2R4 (wherein R4 is
optionally substituted alkyl or aryl) or -O (CH2) 2 R5 (wherein R5 is (un)
substituted heterocyclyl).
2. The compound according to claim 1 wherein ring A including X is pyrrolidine, Ri is phenyl (optionally substituted with methyl, phenyl or methoxy) or -(CH2) benzodioxol, Y is -CONH, R2 is nitro, amino, hydroxyl, halogen, -NHCOR3 (wherein R3 is pyridyl, (tertbutoxycarbonyl)piperidinyl, piperidinyl, phenyl, methyl or trifluoromethyl, -NHSO2R4 (wherein R4 is methyl, phenyl or tolyl) and -0(CH2)2R5 (wherein R5 is morpholinyl).
3. A compound, which is selected from:

- (S)-3-{4-[(Pyridine-4-carbonyl)-amino]-phenyl}-2-[((S)-l-p-tolylcarbamoylpyrrolidine2-carbonyl)-amino]-propionic acid,
- (S)-3-(4-Acetylamino-phenyl)-2-{ [ 1 -(biphenyl-2-ylcarbamoyl)-pyrrolidine-2-carbonyl]-amino}-propionic acid,
(S)-2- {[ 1 -Biphenyl-2-ylcarbamoyl)-pyirolidine-2-carbonyl] -amino} -3-[4-(2,2,2-trifluoro-acetylamino)-phenyl]-propionic acid,
- (S)-2-{[l-(Biphenyl-2-ylcarbamoyl)-pyirolidine-2-carbonyl]-amino}-3-(4-
methanesulfonyl amino-phenyl)-propionic acid,
(S)-3-(4-Benzenesulfonylamino-phenyl)-2-{[l-(biphenyl-2-ylcarbamoyl)-pyrrolidine-2-carbonyl] -amino } -propionic acid,
(S)-2-{[l-(Biphenyl-2-ylcarbamoyl)-pyrrolidine-2-carbonyl]-amino}-3-(4-toluene-4-sulfonyl amino) phenyl)-propionic acid,
- (S)-3-{4-[(Pyridine-4-carbonyl)-amino]-phenyl}-2-[((S)-l-o-tolylcarbamoyl-
pyrrolidine-2-carbonyl)-amino]-propionic acid,
(S)-2- {[S -1 -(2-Methoxy-phenylcarbamoy l)-pyrrolidine-2-carbonyl] -amino } -3 -{4-[(pyridine-4-carbonyl)-amino]-propionic acid,
(S)-2-({(S)-l-[(Benzo[l,3]dioxol-5-ylmethyl)-carbamoyl]-pyrrolidine-2-carbonyl}-amino)-3{4-[(pyridine-4-carbonyl)-amino]-phenyl}-propionic acid,
(S)-2-{[(S)-l-(Biphenyl-2-ylcarbamoyl)-pyrrolidine-2-carbonyl]-amino}-3-{4-[(pyridine-4-carbonyl)-amino]-phenyl}-propionic acid,
(S)-3-(4-Benzoylamino-phenyl)-2-{[l-(S)-(biphenyl-2-ylcarbamoyl)-pyrrolidine-2-carbonyl]-amino}-propionic acid,
- 4-[4-((S)-2-{[(S)-l-(Biphenyl-2-ylcarbamoyl)-pyrrolidine-2-carbonyl]-amino}-2-
carboxy-ethyl)-phenylcarbamoyl]-piperidine-l-carboxylic acid tert-butyl ester,
(S)-2-{[(S)-l-(Biphenyl-2-ylcarbamoyl)-pyrrolidine-2-carbonyl]-amino}-3-(4-nitro-phenyl)-propionic acid,
(S)-3-(4-Amino-phenyl)-2-{ [(S)-1 -(biphenyl-2-ylcarbamoyl)-pyrrolidine-2-carbonyl]-amino} propionic acid,
- 4-{4-[(S)-2-({(S)-l-[(Benzo[l,3]dioxol-5-ylmethyl)carbamoyl]-pyrrolidine-2-carbonyl} -amino)-2-carboxy-ethyl] -phenylcarbamoyl }-piperidine-1 -carboxylic acid tert-butyl ester,
- (S)-2-{[l-(Biphenyl-2-ylcarbamoyl)-pyrrolidine-2-carbonyl]-amino}-3-(4-fluoro-phenyl)-propionic acid,
(S)-2-{[l-(Biphenyl-2-ylcarbamoyl)-pyrrolidine-2-carbonyl]-amino}-3-(4-hydroxy-phenyl)-propionic acid,
(S)-2-{ [ 1 -(Biphenyl-2-ylcarbamoyl)-pyrrolidine-2-carbonyl]-amino}-3-[4-(2-morpholin-4yl-ethoxy)-phenyl}-propionic acid,
and its pharmaceutically acceptable salts, pharmaceutically acceptable solvates,
stereoisomers, tautomers, racemates, regioisomers, prodrugs, metabolites,
polymorphs or N-oxides,
- (S)-2- {[(S)-1 -(Biphenyl-2-ylcarbamoyl)-pyrrolidine-2-carbonyl] -amino} -3 - {4-
[(piperidine-4-carbonyl)-amino]-phenyl}-propionic acid TFA salt,
and its pharmaceutically acceptable solvates, stereoisomers, tautomers, racemates, regioisomers, prodrugs, metabolites, polymorphs or N-oxides.
4. A combination of a compound of any of the preceding claims 1 to 3 with the therapeutic agents selected from corticosteroids, beta agonists, leukotriene antagonists, 5-lipoxygenase inhibitors, phosphodiesterase inhibitors, chemokine inhibitors and muscarinic receptor antagonists.
5. A pharmaceutical composition comprising a therapeutically effective amount of a compound of any one of the proceeding claims together with pharmaceutically acceptable carrier, excipients or diluents.
6. Use of compounds according to any of the preceding claims in the manufacture of medicament for treating an animal or a human suffering from bronchial asthma, rheumatoid arthritis, type I diabetes, multiple sclerosis, psoriasis, allograft rejection or other inflammation and/or autoimmune disorders.
7. Use of compounds according to any of the preceding claims in the manufacture of medicament for preventing, inhibiting or suppressing cell adhesion in an animal or human.
8. A method for the preparation of compounds of Formula XIII and their pharmaceutically acceptable salts, pharmaceutically acceptable solvates, stereoisomers, tautomers, racemates, regioisomers, prodrugs, metabolites, polymorphsor N-oxides,
(Formula Removed)
which method comprises
(a) reacting the compounds of Formula II with compounds of Formula III to give compounds of Formula IV (wherein R6 can be alkyl),
(b) coupling the compounds of Formula IV with compounds of Formula V to give compounds of Formula VI (wherein ring A, X can be the same as defined earlier and P can be a protecting group),
(c) reducing the compounds of Formula VI to give compounds of Formula VII,
(d) coupling the compounds of Formula VII with compounds of Formula VIII (wherein R7 can be hydroxy or halogen) to give compounds of Formula IX ( wherein R3 can be the same as defined earlier),
(e) deprotecting the compounds of Formula IX to give compounds of Formula X,
(f) condensing the compounds of Formula X with compounds of Formula XI to give compounds of Formula XII (wherein R1 can be the same as defined earlier),
(g) hydrolyzing the compounds of Formula XII to give compounds of Formula XIII,
(h) converting the compounds of Formula XIII to their pharmaceutically acceptable salts. 9. A method for the preparation of compounds of Formula XXIV and XXVII and their pharmaceutically acceptable salts, pharmaceutically acceptable solvates, stereoisomers, tautomers, racemates, regioisomers, prodrugs, metabolites, polymorphs
or N-oxides,
(Formula Removed)
which method comprises,
(a) reacting the compounds of Formula XIV with compounds of Formula XV (wherein X can be halogen) to give compounds of Formula XVI (wherein R1 can be the same as defined earlier),
(b) condensing the compounds of Formula XVI with compounds of Formula XVII to give compounds of Formula XVIII (wherein ring A, R6 and X can be the same as defined earlier),
(c) hydrolyzing the compounds of Formula XVIII to give compounds of Formula XIX,
(d) coupling the compounds of Formula XIX with compounds of Formula IV to give compounds of Formula XX,
(e) reducing the compounds of Formula XX to give compounds of Formula XXI,
(f) reacting the compounds of Formula XXI with compounds of Formula XXII to give compounds of Formula XXIII (wherein R4 can be the same as defined earlier), or reacting the compounds of Formula XXI with compounds of Formula XXV to give compounds of Formula XXVI (wherein R3 can be same as defined earlier),
(g) hydrolyzing the compounds of Formula XXIII to give compounds of Formula XXIV.
(h) hydrolyzing the compounds of Formula XXVI to give compounds of Formula
XXVII. (i) converting the compounds of Formula XXIV and XXVII to their
pharmaceutically acceptable salts.
10. A method for the preparation of compounds of Formula XIII, XXXI and XXXII
and their pharmaceutically acceptable salts, pharmaceutically acceptable
solvates, stereoisomers, tautomers, racemates, regioisomers, prodrugs,
metabolites, polymorphs or N-oxides,
(Formula Removed)
which method comprises
(a) deprotecting the compounds of Formula VI to give compounds of Formula
XXVIII (wherein ring A, R6 and X can be the same as defined earlier),
(b) condensing the compounds of Formula XXVIII with compounds of Formula XI
or compounds of Formula XVI to give compounds of Formula XXIX,
(c) reducing the compounds of Formula XXIX to give compounds of Formula XXX or hydrolyzing the compounds of Formula XXIX to give compounds of Formula XXXI,
(d) coupling the compounds of Formula XXX with compounds of Formula VIII ( wherein R7 can be the same as defined earlier) to give compounds of Formula XII (wherein R3 can be the same as defined earlier),
(e) hydrolyzing the compounds of Formula XII to give compounds of Formula XIII.
(f) reducing the compounds of Formula XXXI to give compounds of Formula XXXII.
(g) converting the compounds of Formula XIII, XXXI and XXXII to their
pharmaceutically acceptable salts.
11. A method for the preparation of compounds of Formula XXXV and XXXVIII
and their pharmaceutically acceptable salts, pharmaceutically acceptable solvates, stereoisomers, tautomers, racemates, regioisomers, prodrugs, metabolites, polymorphs or N-oxides,
(Formula Removed)
which method comprises
(a) coupling the compounds of Formula XIX with compounds of Formula XXXIII to
give compounds of Formula XXXIV (wherein ring A, X, R1, R2 and R6 can be the
same as defined earlier) or reacting the compounds of Formula XXXIV with
compounds of Formula XXXVI (wherein X1 can be halogen) to give compounds
of Formula XXXVII (wherein R5 can be the same as defined earlier),
(b) hydrolyzing the compounds of Formula XXXIV to give compounds of Formula
XXXV.
(c) hydrolyzing the compounds of Formula XXXVII to give compounds of Formula
XXXVIII.
(d) converting the compounds of Formula XXXV and XXXVIII to their
pharmaceutically acceptable salts.