Cellular compositions used to restore stem cell or progenitor cell function and methods related thereto

Abstract

This disclosure relates to compounds, compositions and methods of epigenetically transforming cells. In certain embodiments, the disclosure relates to methods of generating epigenetically altered cells comprising mixing isolated cells with compositions disclosed herein under conditions such that epigenetically altered cells are formed. In certain embodiments, methods of treating or preventing vascular or diabetic diseases or conditions are contemplated. In certain embodiments, epigenetically reprogramming adult bone marrow-derived stem or progenitor cells including mesenchymal stem cells (MSCs) or endothelial progenitor cells (EPCs) for autologous treatments are contemplated.

1 . A method of epigenetically modifying progenitor cells comprising mixing stem or progenitor cells and compositions comprising a quinazoline compound under conditions such that cells with enhanced angiogenic gene expression are produced. 
     
     
         2 . The method of  claim 1 , wherein the conditions are such that reduced DNA methylation in the promoters of angiogenic genes occurs. 
     
     
         3 . The method of  claim 1 , wherein the angiogenic genes are one or more or all of the genes selected from Akt1, Hgf, Mapk14, Sphk1, Vegfc, Nudt6, Kdr, Vegfa, and Pten. 
     
     
         4 . The method of  claim 1 , wherein the stem or progenitor cells are mesenchymal stem cells (MSCs) or endothelial progenitor cells (EPCs), cardiac stem cells, myoblasts, adult bone marrow-derived cells, umbilical cord blood cells, hematopoietic stem cells, fibroblasts, or peripheral blood CD34 +  cells. 
     
     
         5 . The method of  claim 4 , wherein the cells endothelial progenitor cells are bone marrow derived. 
     
     
         6 . The method of  claim 5 , wherein the cells are obtained from a subject diagnosed with diabetes and/or cardiovascular disease. 
     
     
         7 . The method of  claim 1 , wherein the composition further comprising a DNA methyltransferase inhibitor, a histone deacetylase (HDAC) inhibitor, DNA methylation inhibitor, a Rho-associated kinase (ROCK) inhibitor, Wnt inhibitor, GSK-3beta inhibitor, or a dihydropyridine. 
     
     
         8 . The method of  claim 7 , wherein the DNA methyltransferase inhibitor is N-phthalyl-L-tryptophan (RG 108). 
     
     
         9 . The method of  claim 7 , wherein the DNA methylation inhibitor is 5-azacitidine or decitabine. 
     
     
         10 . The method of  claim 7 , wherein the HDAC inhibitor is vorinostat-suberoylanilide hydroxamic acid (SAHA), trichostatin A (TSA), or valproic acid (VPA). 
     
     
         11 . The method of  claim 7 , wherein the ROCK inhibitor is 4-(1-aminoethyl)-N-(pyridin-4-yl)cyclohexanecarboxamide (Y-27632) or salt thereof. 
     
     
         12 . The method of  claim 7 , wherein the dihydropyridine is 1,4-dihydro-2,6-dimethyl-5-nitro-4-[2-(trifluoromethyl)phenyl]-3-pyridinecarboxylic acid, methyl ester (BayK8644), ester, derivative, or salt thereof. 
     
     
         13 . The method of  claim 7 , wherein the GSK-3beta inhibitor is 6-[[2-[[4-(2,4-dichlorophenyl)-5-(5-methyl-1H-imidazol-2-yl)-2 pyrimidinyl]amino]ethyl]amino]-3-pyridinecarbonitrile (CHIR99201) or salt thereof. 
     
     
         14 . The method of  claim 1 , wherein the quinazoline compound has formula I, 
       
         
           
           
               
               
           
         
         or salts thereof wherein, 
         R 1 , R 2 , R 3 , R 4 , R 5 , and R 6  are each the same or different hydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl) 2 amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, 
         wherein each R 1 , R 2 , R 3 , R 4 , R 5 , and R 6  are optionally substituted with one or more, the same or different, R 7 ; 
         R 7  is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl) 2 amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R 7  is optionally substituted with one or more, the same or different, R 8 ; and 
         R 8  is halogen, nitro, cyano, hydroxy, trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy, acetyl, acetoxy, methylamino, ethylamino, dimethylamino, diethylamino, N-methyl-N-ethylamino, acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio, ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl, methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl. 
       
     
     
         15 . The method of  claim 14 , wherein the quinazoline compound has Formula IA: 
       
         
           
           
               
               
           
         
         or salts thereof wherein, 
         R 1 , R 3 , R 4 , R 5 , and R 6  are each the same or different hydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl) 2 amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein each R 1 , R 2 , R 3 , R 4 , R 5 , and R 6  are optionally substituted with one or more, the same or different, R 7 ; 
         R 7  is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl) 2 amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R 7  is optionally substituted with one or more, the same or different, R 8 ; 
         R 8  is halogen, nitro, cyano, hydroxy, trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy, acetyl, acetoxy, methylamino, ethylamino, dimethylamino, diethylamino, N-methyl-N-ethylamino, acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio, ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl, methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl; 
         R 9  and R 10  are each the same or different hydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl) 2 amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein each R 9  and R 10  are optionally substituted with one or more, the same or different, R 11 ;
 or R 9  and R 10  come together to form a heterocyclyl optionally substituted with one or more, the same or different, R 11 ; 
 
         R 11  is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl) 2 amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R 11  is optionally substituted with one or more, the same or different, R 12 ; and 
         R 12  is halogen, nitro, cyano, hydroxy, trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy, acetyl, acetoxy, methylamino, ethylamino, dimethylamino, diethylamino, N-methyl-N-ethylamino, acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio, ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl, methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl. 
       
     
     
         16 . The method of  claim 1 , wherein the quinazoline compound is selected from:
 N-(1-benzylpiperidin-4-yl)-2-(4-methylpiperazin-1-yl)quinazolin-4-amine,   N-(1-benzylpiperidin-4-yl)-2-(4-methyl-1,4-diazepan-1-yl)quinazolin-4-amine,   N-(1-benzylpiperidin-4-yl)-6,7-dimethoxy-2-(4-methyl-1,4-diazepan-1-yl)quinazolin-4-amine,   N-(1-benzylpiperidin-4-yl)-6,7-dimethoxy-2-(4-methylpiperazin-1-yl)quinazolin-4-amine,   N-(1-benzylpiperidin-4-yl)-6,7-dimethoxy-2-morpholinoquinazolin-4-amine,   N-(1-benzylpiperidin-4-yl)-2-morpholinoquinazolin-4-amine,   N-(1-benzylpiperidin-4-yl)-2-thiomorpholinoquinazolin-4-amine,   N-(1-benzylpiperidin-4-yl)-2-(piperidin-1-yl)quinazolin-4-amine,   2-(azepan-1-yl)-N-(1-benzylpiperidin-4-yl)quinazolin-4-amine,   N-(1-benzylpiperidin-4-yl)-2-(4-methylpiperidin-1-yl)quinazolin-4-amine,   N 4 -(1-benzylpiperidin-4-yl)-N 2 -(piperidin-1-yl)quinazoline-2,4-diamine,   N-(1-benzylpiperidin-4-yl)-2-(pyrrolidin-1-yl)quinazolin-4-amine,   N-(1-benzylpiperidin-4-yl)-2-(4-(pyridin-2-yl)piperazin-1-yl)quinazolin-4-amine,   (4-(4-((1-benzylpiperidin-4-yl)amino)quinazolin-2-yl)piperazin-1-yl)(phenyl)methanone,   N 4 -(1-benzylpiperidin-4-yl)-N 2 -(3-(dimethylamino)propyl)quinazoline-2,4-diamine,   2-((4-((1-benzylpiperidin-4-yl)amino)quinazolin-2-yl)amino)ethanol,   N 4 -(1-benzylpiperidin-4-yl)-N 2 -(4-methylpiperazin-1-yl)quinazoline-2,4-diamine,   N 4 -(1-benzylpiperidin-4-yl)-N 2 -morpholinoquinazoline-2,4-diamine,   N-(1-benzylpiperidin-4-yl)-2-(2-methylpiperidin-1-yl)quinazolin-4-amine,   7-((5-aminopentyl)oxy)-N 4 -(1-benzylpiperidin-4-yl)-N 2 -(3-(dimethylamino)propyl)-6-methoxyquinazoline-2,4-diamine,   N-(1-benzylpiperidin-4-yl)-2-(4-phenylpiperazin-1-yl)quinazolin-4-amine, and   7-((5-aminopentyl)oxy)-N 4 -(1-(5-aminopentyl)piperidin-4-yl)-N 2 -(3-(dimethylamino)propyl)-6-methoxyquinazoline-2,4-diamine or salts thereof.   
     
     
         17 . The method of  claim 1 , wherein the quinazoline compound has Formula II: 
       
         
           
           
               
               
           
         
         or salts thereof wherein, 
         R 2 , R 3 , R 4 , R 5 , and R 6  are each the same or different hydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl) 2 amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein each R 2 , R 3 , R 4 , R 5 , and R 6  are optionally substituted with one or more, the same or different, R 7 ; 
         R 7  is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl) 2 amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R 7  is optionally substituted with one or more, the same or different, R 8 ; 
         R 8  is halogen, nitro, cyano, hydroxy, trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy, acetyl, acetoxy, methylamino, ethylamino, dimethylamino, diethylamino, N-methyl-N-ethylamino, acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio, ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl, methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl; 
         R 13  and R 14  are each the same or different hydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl) 2 amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein each R 13  and R 14  are optionally substituted with one or more, the same or different, R 15 ; 
         R 15  is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, alkanoyl, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl) 2 amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R 15  is optionally substituted with one or more, the same or different, R 16 ; and 
         R 16  is halogen, nitro, cyano, hydroxy, trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy, acetyl, acetoxy, methylamino, ethylamino, dimethylamino, diethylamino, N-methyl-N-ethylamino, acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio, ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl, methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl. 
       
     
     
         18 . The method of  claim 17 , wherein the quinazoline compound is selected from:
 7-((5-aminopentyl)oxy)-N 2 -(3-(dimethylamino)propyl)-6-methoxy-N 4 ,N 4 -dimethylquinazoline-2,4-diamine and   7-((5-aminopentyl)oxy)-6-methoxy-N 2 ,N 2 ,N 4 ,N 4 -tetramethylquinazoline-2,4-diamine or salts thereof.   
     
     
         19 . A method of treating or preventing vascular disease or condition comprising:
 mixing progenitor cells from a subject and a composition as provided in  claim 1 , under conditions such that epigenetically modified cells with enhanced angiogenic gene expression are produced; and   administering an effective amount of a composition comprising the epigenetically modified cells or cells cultured therefrom to a subject in need thereof.   
     
     
         20 . The method of  claim 19 , wherein the progenitor cells are bone marrow derived cells, endothelial progenitor cells, or mesenchymal stem cells. 
     
     
         21 . The method of  claim 20 , wherein the progenitor cells were obtained from the subject receiving the administered composition. 
     
     
         22 . The method of  claim 20 , wherein the vascular disease or condition is peripheral vascular disease, myocardial ischemia, heart failure, or stroke. 
     
     
         23 . A method of treating or preventing a diabetic disease or conditions comprising:
 mixing progenitor cells from a subject and a composition as provided in  claim 1  under conditions such that epigenetically modified cells with enhanced angiogenic gene expression are produced; and   administering an effective amount of a composition comprising the epigenetically modified cells or cells cultured therefrom to a subject in need thereof.   
     
     
         24 . The method of  claim 23 , wherein the progenitor cells are bone marrow derived cells, endothelial progenitor cells, or mesenchymal stem cells. 
     
     
         25 . The method of  claim 24 , wherein the progenitor cells were obtained from the subject receiving the administered composition. 
     
     
         26 . The method of  claim 23 , wherein the diabetic disease or condition is diabetic wounds or diabetic neuropathy.