Agonists that enhance binding of integrin-expressing cells to integrin receptors

Abstract

A method of enhancing binding of cells to an integrin-binding ligand comprises treating integrin-expressing cells in vitro with an agonist of integrin, wherein the integrin is selected from the group consisting of α4fβ1, α5β1, α4β7, αvβ3 and αLβ2, and contacting the treated cells with an integrin-binding ligand; integrin agonist compounds having the general formula I; methods of treating integrin-expressing cells with such agonists to enhance binding; and therapeutic methods comprising administering agonist-treated cells or agonist compounds to a mammal.

What is claimed is: 
     
         1 . A chemical compound having the general formula (I) 
       
         
           
           
               
               
           
         
         wherein 
         R 1  is alkyl, aryl or aralkyl, 
         R 2  is selected from the group consisting of aralkyl and alkyl, provided that when R 1  is alkyl, R 2  is aralkyl, 
         M 1  is CO or SO 2 ,
 provided that when M 1  is SO 2  and R 1  is phenyl, 4-methylphenyl or 2,4,6-trimethylphenyl, R 2  is not alkyl, 2-phenethyl, benzyl, or 2-methoxy-2-oxoethyl, and when M 1  is CO and R 1  is 2-furyl, 4-pyridyl, or 3,5-dinitrophenyl, R 2  is not alkyl, benzyl or 2-(1H-indol-2-yl)ethyl, 
 
         M 2  is absent or CH 2 , 
         M 3  and M 4  are absent, 
         M 5  is (CR 11 R 12 ), 
         R 11  is hydrogen, 
         R 12  is selected from the group consisting of hydrogen, NR 21 CONR 22 R 23 , NR 21 COR 24 , NR 21 SO 2 R 24 , NR 21 COOR 24 , CONR 22 R 23 , COOR 24 , O(CH 2 CH 2 O) s R 24  hydroxyalkyl and alkoxyalkyl, wherein s is an integer of 1 to 6, 
         M 6  is (CH 2 ) q  where q is an integer of 0 to 6, 
         R 3  is selected from the group consisting of NR 15 COOR 16 , NR 15 COR 16 ,NR 15 CONR 13 R 14 , and NR 15 SO 2 R 16 , and 
         R 13 , R 21  and R 22 , when present, are independently selected from the group consisting of hydrogen and C 1 -C 6  alkyl, and 
         R 14 , R 15  R 16 , R 23  and R 24 , each of which when present, is independently selected from the group consisting of hydrogen, alkyl, aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl and aralkyl, and 
         R 1 , R 2 , R 3 , R 12 , R 14 , R 15 , R 16 , R 23  and R 24 , when present, may independently be either unsubstituted or substituted with one or more substituents selected from the group consisting of alkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylaryl, hydroxy, alkoxy, azido, haloalkoxy, hydroxyalkyl, aryloxy, hydroxyaryl, alkoxyaryl, halo, haloalkyl, haloaryl, amino, alkylamino, dialkylamino, arylamino, diarylamino, —NHCO(alkyl), —NHCO(aryl), —NHCO(aralkyl), —NHCO(haloalkyl), —NHSO 2 (alkyl), —NHSO 2 (aryl), —NHSO 2 (aralkyl), alkoxycarbonyl, alkoxycarbonylalkyl, —OCO(alkylamino) and —OCO(dialkylamino), wherein the term “aryl” refers to a carbocyclic aromatic group having 6 to 12 carbon atoms or a heterocyclic aromatic group, and 
         wherein the term “aralkyl” refers to an aryl substituted alkyl radical, wherein the term “aryl” is as defined above. 
       
     
     
         2 . A compound of  claim 1  selected from the group consisting of methyl (2S)-6-{[(benzyloxy)carbonyl]amino}-2-[benzyl(2-thienylsulfonyl)amino]hexanoate; methyl (2S)-6-{[(benzyloxy)carbonyl]amino}-2-[benzyl(phenylsulfonyl)amino]hexanoate; methyl (2S)-6-{[(benzyloxy)carbonyl]amino}-2-[(2-thienylcarbonyl)(2-thienylmethyl)amino]hexanoate; methyl (2S)-6-{[(benzyloxy)carbonyl]amino}-2-[(2-thienylacetyl)(2-thienylmethyl)amino]hexanoate; methyl (2S)-2-[benzyl(isobutylsulfonyl)amino]-6-{[(benzyloxy)carbonyl]amino}hexanoate; benzyl {(5S)-5-[(tert-butoxycarbonyl)amino]-6-[(2-thienylmethyl)(2-thienylsulfonyl)amino]hexyl}carbamate; benzyl {(5S)-5-[(tert-butoxycarbonyl)amino]-6-[(2-thienylacetyl)(2-thienylmethyl)amino]hexyl}carbamate; benzyl {(5S)-5-[(tert-butoxycarbonyl)amino]-6-[(methyl sulfonyl)(2-thienylmethyl)amino]hexyl}carbamate; benzyl {(5S)-5-[(tert-butoxycarbonyl)amino]-6-[(phenylsulfonyl)(2-thienylmethyl)amino]hexyl}carbamate; benzyl {(5S)-5-[(tert-butoxycarbonyl)amino]-6-[(2-thienylcarbonyl)(2-thienylmethyl)amino]hexyl}carbamate; N,N′-heptane-1,7-diylbis[N-(2-thienylmethyl)benzamide]; N,N′-heptane-1,7-diylbis[N-(2-thienylmethyl)thiophene-2-carboxamide]; benzyl [(5S)-5-[(tert-butoxycarbonyl)amino]-6-{[(4-methoxyphenyl)sulfonyl](2-thienylmethyl)amino}hexyl]carbamate; benzyl {(5S)-5-[(tert-butoxycarbonyl)amino]-6-[(4-methoxybenzoyl)(2-thienylmethyl)amino]hexyl}carbamate; N,N′-hexane-1,6-diylbis[N-(2-thienylmethyl)thiophene-2-carboxamide]; N,N′-hexane-1,6-diylbis[N-(3-methoxybenzyl)thiophene-2-carboxamide]; tert-butyl {5-[(4-methoxybenzyl)(2-thienylsulfonyl)amino]pentyl}carbamate; N,N′-pentane-1,5-diylbis[N-(3-methoxybenzyl)thiophene-2-sulfonamide]; N-(3-methoxybenzyl)-N-{5-[(2-thienylsulfonyl)amino]pentyl}thiophene-2-sulfonamide; tert-butyl {5-[(2-thienylcarbonyl)(2-thienylmethyl)amino]pentyl}carbamate; N-(3-methoxybenzyl)-N-{5-[(2-thienylcarbonyl)amino]pentyl}thiophene-2-carboxamide; and N,N′-pentane-1,5-diylbis[N-(3-methoxybenzyl)thiophene-2-carboxamide]. 
     
     
         3 . A chemical compound of  claim 1  having the general formula (I) 
       
         
           
           
               
               
           
         
         wherein 
         R 1  is alkyl, aryl or aralkyl, 
         R 2  is selected from the group consisting of arylmethyl, arylethyl and alkyl, provided that when R 1  is alkyl, R 2  is arylmethyl, or arylethyl 
         M 1  is CO or SO 2 ,
 provided that when M 1  is SO 2  and R 1  is phenyl, 4-methylphenyl or 2,4,6-trimethylphenyl, R 2  is not alkyl, 2-phenethyl, benzyl, or 2-methoxy-2-oxoethyl, and when M 1  is CO and R 1  is 2-furyl, 4-pyridyl, or 3,5-dinitrophenyl, R 2  is not alkyl, benzyl or 2-(1H-indol-2-yl)ethyl, 
 
         M 2  is absent or CH 2 , 
         M 3  and M 4  are absent, 
         M 5  is (CR 11 R 12 ), 
         R 11  is hydrogen, 
         R 12  is selected from the group consisting of hydrogen, NR 21 COOR 24 , COOR 24 , 
         M 6  is (CH 2 ) q  where q is an integer of 0 to 6, 
         R 3  is selected from the group consisting of NR 15 COOR 16 , NR 15 COR 16  and NR 15 SO 2 R 16 , and 
         R 21  when present is selected from the group consisting of hydrogen and C 1 -C 6  alkyl, and 
         R 15 , R 16 and R 24 , each of which, when present is independently selected from the group consisting of hydrogen, alkyl, aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl and aralkyl, and 
         R 1 , R 2 , R 3 , R 12 , R 15 , R 16  and R 24  when present may independently be either unsubstituted or substituted with one or more substituents selected from the group consisting of alkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylaryl, hydroxy, alkoxy, azido, haloalkoxy, hydroxyalkyl, aryloxy, hydroxyaryl, alkoxyaryl, halo, haloalkyl, haloaryl, amino, alkylamino, dialkylamino, arylamino, diarylamino, —NHCO(alkyl), —NHCO(aryl), —NHCO(aralkyl), —NHCO(haloalkyl), —NHSO 2 (alkyl), —NHSO 2 (aryl), —NHSO 2 (aralkyl), alkoxycarbonyl, alkoxycarbonylalkyl, —OCO(alkylamino) and —OCO(dialkylamino). 
       
     
     
         4 . A chemical compound of  claim 3  having the general formula (I) 
       
         
           
           
               
               
           
         
         wherein 
         R 1  is C 1 -C 6  alkyl, arylmethyl, arylethyl, phenyl, thienyl, furyl, pyridyl, thiazolyl, oxazolyl, isoxazolyl or pyrrolyl, 
         R 2  is selected from the group consisting of arylmethyl, arylethyl and alkyl, provided that when R 1  is C 1 -C 6  alkyl, R 2  is arylmethyl or arylethyl, 
         M 1  is CO or SO 2 ,
 provided that when M 1  is SO 2  and R 1  is phenyl, 4-methylphenyl or 2,4,6-trimethylphenyl, R 2  is not alkyl, 2-phenethyl, benzyl, or 2-methoxy-2-oxoethyl, and when M 1  is CO and R 1  is 2-furyl, 4-pyridyl, or 3,5-dinitrophenyl, R 2  is not alkyl, benzyl or 2-(1H-indol-2-yl)ethyl, 
 
         M 2  is absent or CH 2 , 
         M 3  and M 4  are absent, 
         M 5  is (CR 11 R 12 ), 
         R 11  is hydrogen, 
         R 12  is selected from the group consisting of hydrogen, NR 21 COOR 24 , COOR 24 , 
         M 6  is (CH 2 ) q  where q is an integer of 0 to 6, 
         R 3  is selected from the group consisting of NR 15 COOR 16 , NR 15 COR 16  and NR 15 SO 2 R 16 , and 
         R 21  when present is selected from the group consisting of hydrogen and C 1 -C 6  alkyl, and 
         R 15 , R 16  and R 24 , each of which, when present is independently selected from the group consisting of hydrogen, alkyl, aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl and aralkyl, and 
         R 1 , R 2 , R 3 , R 12 , R 15 , R 16 , and R 24  when present may independently be either unsubstituted or substituted with one or more substituents selected from the group consisting of alkyl, aryl, aralkyl, hydroxy, alkoxy, azido, haloalkoxy, hydroxyalkyl, hydroxyaryl, alkoxyaryl, halo, amino, alkylamino, dialkylamino, NHCO(alkyl), —NHCO(aryl), —NHCO(aralkyl), —NHCO(haloalkyl), —NHSO 2 (alkyl), —NHSO 2 (aryl), —NHSO 2 (aralkyl), alkoxycarbonyl. 
       
     
     
         5 . A chemical compound of  claim 4  having the general formula (I) 
       
         
           
           
               
               
           
         
         wherein 
         R 1  is phenyl, thienyl, furyl, pyridyl, thiazolyl, oxazolyl, isoxazolyl or pyrrolyl, 
         R 2  is selected from the group consisting of arylmethyl, or arylethyl, M 1  is CO or SO 2 ,
 provided that when M 1  is SO 2  and R 1  is phenyl, 4-methylphenyl or 2,4,6-trimethylphenyl, R 2  is not 2-phenethyl or benzyl, and when M 1  is CO and R 1  is 2-furyl, 4-pyridyl, or 3,5-dinitrophenyl, R 2  is not benzyl or 2-(1H-indol-2-yl)ethyl, 
 
         M 2  is absent or CH 2 , 
         M 3  and M 4  are absent, 
         M 5  is (CR 11 R 12 ), 
         R 11  is hydrogen, 
         R 12  is hydrogen, 
         R 3  is selected from the group consisting of NR 15 COR 16 , and NR 15 SO 2 R 16 , and 
         R 15  is hydrogen or aralkyl 
         R 16  is phenyl, thienyl, furyl, pyridyl, thiazolyl, oxazolyl, isoxazolyl or pyrrolyl, 
         R 1 , R 2 , R 3 , R 12 , R 15  and R 16  when present may independently be either unsubstituted or substituted with one or more substituents selected from the group consisting of alkyl, aryl, aralkyl, hydroxy, alkoxy, azido, haloalkoxy, hydroxyalkyl, hydroxyaryl, alkoxyaryl, halo, amino, alkylamino, dialkylamino, NHCO(alkyl), —NHCO(aryl), —NHCO(aralkyl), —NHCO(haloalkyl), —NHSO 2 (alkyl), —NHSO 2 (aryl), —NHSO 2 (aralkyl), alkoxycarbonyl. 
       
     
     
         6 . A pharmaceutical composition comprising:
 a compound of  claim 1  or a pharmaceutically acceptable salt thereof; and   a pharmaceutically acceptable carrier.   
     
     
         7 . A method of treating integrin-expressing cells, comprising:
 contacting at least one integrin-expressing cell in vitro with an agonist of said integrin, wherein said agonist is a compound of  claim 1 .   
     
     
         8 . The method of  claim 7 , wherein said integrin is selected from the group consisting of α4β1, α5β1, α4β7, αvβ3 and  Lβ2. 
     
     
         9 . A method of enhancing binding of cells to an integrin-binding ligand, comprising:
 treating integrin-expressing cells in vitro with an agonist of integrin according to the method of  claim 8 , wherein said integrin is selected from the group consisting of α4β1, α5β1, α4β7, αvβ3 and αLβ2; and   contacting the treated cells with an integrin-binding ligand.   
     
     
         10 . The method of  claim 9 , wherein said integrin-expressing cells are selected from the group consisting of adult stem cells, embryonic stem cells, progenitor cells, and induced pluripotent stem cells. 
     
     
         11 . The method of  claim 9 , wherein contacting the treated cells with an integrin-binding ligand includes contacting a surface comprising an integrin-binding ligand with the agonist-treated cells, to bind the agonist-treated cells to said surface, wherein binding of said agonist-treated cells is enhanced relative to binding of integrin-expressing cells not treated by this method. 
     
     
         12 . The method of  claim 11 , wherein at least 3 fold more agonist-treated cells are bound to said surface than integrin-expressing cells not treated with said agonist. 
     
     
         13 . The method of  claim 11 , wherein said surface is on a tissue comprising an integrin binding protein selected from the group consisting of vascular cell adhesion molecule-1 (VCAM-1), fibronectin, mucosal addressin cellular adhesion molecule-1 (MAdCAM-1), inter-cellular adhesion molecule-1 (ICAM-1), inter-cellular adhesion molecule-2 (ICAM-2) and vitronectin. 
     
     
         14 . An agonist of integrin for use in a method of enhancing retention of exogenously-introduced cells at an in vivo target site in a mammal, the method comprising:
 (a) treating integrin-expressing cells in vitro with said agonist of integrin;   (b) introducing the agonist-treated cells to an in vivo target site in the mammal; and   (c) causing a greater number of said introduced agonist-treated cells to remain at said target site relative to the number of cells retained if integrin-expressing cells not treated with said agonist were introduced to said target site;   wherein said agonist is a compound as defined in any one of  claim 1 .   
     
     
         15 . Agonist-treated integrin-expressing stem cells or progenitor cells for use in a method of treating damaged or diseased vascular tissue of a mammal, the method comprising:
 (a) administering to a damaged or diseased vascular site, including bone marrow, in a vessel of the mammal a plurality of said agonist-treated integrin-expressing stem cells or progenitor cells;   (b) causing a greater number of said administered agonist-treated cells to remain at said vascular site relative to the number of cells retained if integrin-expressing cells not treated with said agonist were administered to said damaged or diseased vascular site, wherein said site comprises cells bearing an integrin-binding ligand on a cell surface; and   (c) allowing said cells at said vascular site to proliferate/differentiate and/or release paracrine factors;   
       wherein said agonist-treated integrin-expressing stem cells or progenitor cells have been treated according to the method of  claim 7 , said integrin selected from the group consisting of α4β1, α5β1, α4β7, αvβ3 and αLβ2.