Solubilized compositions for controlled proliferation of stem cells / generating inner ear hair cells using gsk3 inhibitors: i

Abstract

The present invention relates to compositions and methods of inducing the self-renewal of stem/progenitor supporting cells, including inducing the stem/progenitor cells to proliferate while maintaining, in the daughter cells, the capacity to differentiate into hair cells.

1 . A pharmaceutical composition comprising:
 a) a pharmaceutically acceptable salt of a 2-pyrimidinylaminoethylamino-2-pyridinyl containing compound;   and   b) a poloxamer;   wherein the pH of the composition is between about 5 and about 9; and   wherein the solubility of the pharmaceutically acceptable salt of the 2-pyrimidinylaminoethylamino-2-pyridinyl containing compound in the pharmaceutical composition is 3-fold higher than the solubility of the pharmaceutically acceptable salt of the 2-pyrimidinylaminoethylamino-2-pyridinyl containing compound in the same composition at the same pH in the absence poloxamer.   
     
     
         2 . The composition of  claim 1 , wherein 2-pyrimidinylaminoethylamino-2-pyridine containing compound is a 2,4-dichlorophenyl-5-(1H-imidazol-2-yl)-2-pyrimidinylaminoethylamino-3-pyridine containing compound. 
     
     
         3 . The composition of  claim 2 , wherein the 2-pyrimidinylaminoethylamino-2-pyridine containing compound is 6-[[2-[[4-(2,4-Dichlorophenyl)-5-(5-methyl-1H-imidazol-2-yl)-2-pyrimidinyl]amino]ethyl]amino]-3-pyridinecarbonitrile, or a pharmaceutically acceptable salt thereof, having a Formula I: 
       
         
           
           
               
               
           
         
       
     
     
         4 . The composition of  claim 1 , wherein the poloxamer comprises at least one of poloxamer 124, Poloxamer 188, Poloxamer 237, Poloxamer 338 or Poloxamer 407. 
     
     
         5 . The composition of  claim 1 , wherein the poloxamer comprises mixtures of two or more of Poloxamer 124, Poloxamer 188, Poloxamer 237, Poloxamer 338 or Poloxamer 407. 
     
     
         6 . The composition of  claim 5 , wherein the mixture of two or more poloxamers comprises Poloxamer 407 and Poloxamer 124. 
     
     
         7 . The composition of  claim 1 , wherein the poloxamer comprises at least one of Poloxamer 188 and Poloxamer 407 or mixtures thereof. 
     
     
         8 . The composition of  claim 1 , wherein the poloxamer is Poloxamer 407. 
     
     
         9 . The composition of  claim 1 , wherein the poloxamer is at a concentration between about 5 wt % and about 25 wt % relative to the composition. 
     
     
         10 . The composition of  claim 1 , wherein the poloxamer is at a concentration between about 10 wt % and about 23 wt % relative to the composition. 
     
     
         11 . The composition of  claim 1 , wherein the poloxamer is at a concentration between about 15 wt % and about 20 wt % relative to the composition. 
     
     
         12 . The composition of  claim 1 , wherein the poloxamer is at a concentration of approximately 17 wt % relative to the composition. 
     
     
         13 . A pharmaceutical composition comprising:
 a) a pharmaceutically acceptable salt of the compound of the Formula I:   
       
         
           
           
               
               
           
         
         and 
         b) a poloxamer; 
         wherein the pH of the composition is between about 5 and about 9; and 
         wherein the solubility of the pharmaceutically acceptable salt of the compound of Formula I in the pharmaceutical composition is 3-fold higher than the solubility of the pharmaceutically acceptable salt of the compound of Formula I in the same composition at the same pH in the absence of poloxamer. 
       
     
     
         14 . The composition of  claim 13 , wherein the poloxamer comprises at least one of Poloxamer 124, Poloxamer 188, Poloxamer 237, Poloxamer 338 or Poloxamer 407. 
     
     
         15 . The composition of  claim 13 , wherein the poloxamer comprises mixtures of two or more of Poloxamer 124, Poloxamer 188, Poloxamer 237, Poloxamer 338 or Poloxamer 407. 
     
     
         16 . The composition of  claim 15 , wherein the mixture of two or more poloxamers comprises Poloxamer 407 and Poloxamer 124. 
     
     
         17 . The composition of  claim 13 , wherein the poloxamer comprises at least one of Poloxamer 188 and Poloxamer 407 or mixtures thereof. 
     
     
         18 . The composition of  claim 13 , wherein the poloxamer is Poloxamer 407. 
     
     
         19 . The composition of  claim 13 , wherein the poloxamer is at a concentration between about 5 wt % and about 25 wt % relative to the composition. 
     
     
         20 . The composition of  claim 13 , wherein the poloxamer is at a concentration between about 10 wt % and about 23 wt % relative to the composition. 
     
     
         21 . The composition of  claim 13 , wherein the poloxamer is at a concentration between about 15 wt % and about 20 wt % relative to the composition. 
     
     
         22 . The composition of  claim 13 , wherein the poloxamer is at a concentration of approximately 17 wt % relative to the composition. 
     
     
         23 . A method of expanding a population of cochlear cells in a cochlear tissue with a stem cell proliferator comprising a parent population of cells, said method comprising contacting the cochlear tissue with a composition of  claim 1  or  13 . 
     
     
         24 . The method of  claim 23 , wherein the stem cell proliferator is capable in a stem cell proliferation assay of increasing the number of Lgr5 +  cells in a stem cell proliferation assay cell population by a factor of at least 10. 
     
     
         25 . The method of  claim 23 , wherein the stem cell proliferator is capable in a stem cell differentiation assay of forming hair cells from a cell population comprising Lgr5 +  cells. 
     
     
         26 . The method of  claim 23 , wherein the cochlear tissue maintains Native Morphology. 
     
     
         27 . The method of  claim 23 , wherein the cochlear tissue is in a subject. 
     
     
         28 . The method of  claim 27 , wherein the contacting the cochlear tissue with the composition is achieved by administering the composition trans-tympanically to the subject. 
     
     
         29 . The method of  claim 27 , wherein contacting the cochlear tissue with the composition results in improved auditory functioning of the subject. 
     
     
         30 . A method of facilitating the generation of tissue cells, the method comprising administering or causing to be administered to a stem cell population a composition of  claim 1  or  13 . 
     
     
         31 . The method of  claim 30 , wherein the tissue cells are cochlear cells. 
     
     
         32 . The method of  claim 30 , wherein the tissue cells are inner ear hair cells. 
     
     
         33 . A method of treating a subject who has, or is at risk of developing, a disease associated with absence or lack of certain tissue cells, comprising administering or causing to be administered to said subject a composition of  claim 1  or  13 . 
     
     
         34 . The method of  claim 33 , wherein the tissue cells are cochlear cells. 
     
     
         35 . The method of  claim 33 , wherein the tissue cells are inner ear hair cells. 
     
     
         36 . A method of treating a subject who has, or is at risk of developing, hearing loss, the method comprising administering to the subject a composition of  claim 1  or  13 . 
     
     
         37 . The method of  claim 36 , wherein the composition is dispersed in a biocompatible matrix. 
     
     
         38 . The method of  claim 37 , wherein the biocompatible matrix is a biocompatible gel or foam. 
     
     
         39 . The method of  claim 36 , wherein the composition is administered trans-tympanically to a cochlear tissue of the subject. 
     
     
         40 . A system for treating a subject who has, or is at risk of developing, a disease associated with absence or lack of certain tissue cells, comprising administering:
 a pharmaceutical composition of  claim 1  or  13 ; and   a trans-tympantic administrative device.   
     
     
         41 . The pharmaceutical composition of  claim 1  further comprising a Notch agonist or HDAC inhibitor. 
     
     
         42 . The pharmaceutical composition of  claim 41 , wherein the differentiation inhibitor is valproic acid. 
     
     
         43 . The pharmaceutical composition of  claim 13  further comprising a Notch agonist or HDAC inhibitor. 
     
     
         44 . The pharmaceutical composition of  claim 43 , wherein the differentiation inhibitor is valproic acid.