Promoter driven tissue specific cytotoxic agents and title methods of use

Abstract

This invention relates to a recombinant nucleic acid for an RIP-tk (rat insulin promoter-thymidine kinase) construct that selectively targets insulin secreting cells, such as β-cells, PDX-1 positive human pancreatic ductal carcinomas, and other cells containing certain transcription factors. The present invention is useful in the treatment of pancreatic cancers, such as β-cell insulinomas and can also be used to target pancreatic tumors that do not express insulin, such as pancreatic adenocarcinoma.

1 - 118 . (canceled)  
     
     
         119 . A method of killing a pancreatic tumor cell in a subject, the method comprising: 
 a) administering to a subject a nucleic acid comprising a vector with an insulin promoter having SEQ ID NO:1 operatively coupled to a cytotoxic gene, wherein the cytotoxic gene is thereby expressed in a pancreatic tumor cell that does not express insulin,    b) administering a pro-drug to said subject, wherein the prodrug is converted to a cytotoxic compound by the action of the protein encoded by said cytotoxic gene and thereby killing the pancreatic tumor cell that does not express insulin.    
     
     
         120 . The method of  claim 119 , where the cytotoxic gene is the thymidine kinase gene.  
     
     
         121 . The method of  claim 119 , where the cytotoxic gene is the thymidine kinase gene and the prodrug is acyclovir, ganciclovir, FIAU or 6-methoxypurine arabinoside.  
     
     
         122 . The method of  claim 121 , wherein the administration is systemic.  
     
     
         123 . The method of  claim 121 , wherein the administration is by direct administration at the site of the pancreatic tumor cell.  
     
     
         124 . A method of treating pancreatic tumor cells in a subject, the method comprising: 
 a) administering to a subject a nucleic acid comprising a vector with an insulin promoter having SEQ ID NO:1 operatively coupled to a cytotoxic gene, wherein the cytotoxic gene is thereby expressed in a PDX-1 positive pancreatic tumor cell,    b) administering a pro-drug to said subject, wherein the prodrug is converted to a cytotoxic compound by the action of the protein encoded by said cytotoxic gene and thereby killing the PDX-1 positive pancreatic tumor cell.    
     
     
         125 . The method of  claim 123 , where the cytotoxic gene is the thymidine kinase gene.  
     
     
         126 . The method of  claim 123 , where the cytotoxic gene is the thymidine kinase gene and the prodrug is acyclovir, ganciclovir, FIAU or 6-methoxypurine arabinoside.  
     
     
         127 . A method of killing a pancreatic tumor cell in a subject, the method comprising: 
 a) administering to a subject a nucleic acid comprising a vector with an insulin promoter having SEQ ID No:1 operatively coupled to a cytotoxic gene, wherein the cytotoxic gene is thereby expressed in a pancreatic tumor cell,    b) administering a pro-drug to said subject, wherein the prodrug is converted to a cytotoxic compound by the action of the protein encoded by said cytotoxic gene and thereby killing the pancreatic tumor cell.    
     
     
         128 . The method of  claim 127 , where the cytotoxic gene is the thymidine kinase gene.  
     
     
         129 . The method of  claim 127 , where the cytotoxic gene is the thymidine kinase gene and the prodrug is acyclovir, ganciclovir, FIAU or 6-methoxypurine arabinoside.  
     
     
         130 . The method of  claim 129 , wherein the administration is systemic.  
     
     
         131 . The method of  claim 129 , wherein the administration is by direct administration at the site of the pancreatic tumor cell.  
     
     
         132 . A method of killing a tumor cell in a subject, the method comprising: 
 a) administering to a subject with a tumor cell expressing PDX-1, a nucleic acid comprising an adenoviral vector with an insulin promoter having SEQ ID NO:1 operatively coupled to a cytotoxic gene, wherein the cytotoxic gene is thereby expressed in the tumor cell expressing PDX-1,    b) administering a pro-drug to said subject, wherein the prodrug is converted to a cytotoxic compound by the action of the protein encoded by said cytotoxic gene and thereby killing the tumor cell expressing PDX-1.    
     
     
         133 . The method of  claim 132 , where the cytotoxic gene is the thymidine kinase gene.  
     
     
         134 . The method of  claim 132 , where the cytotoxic gene is the thymidine kinase gene and the prodrug is acyclovir, ganciclovir, FIAU or 6-methoxypurine arabinoside.  
     
     
         135 . The method of  claim 132 , wherein the administration is systemic.  
     
     
         136 . A method of killing a tumor cell in a subject, the method comprising: 
 a) administering to a subject with a tumor cell expressing PDX-1 a nucleic acid comprising a vector with an insulin promoter, said insulin promoter comprising multiple copies of SEQ ID NO:2 operatively coupled to multiple copies of SEQ ID NO:3 or 4, said insulin promoter operatively coupled to a cytotoxic gene, wherein the cytotoxic gene is thereby expressed in the tumor cell expressing PDX-1,    b) administering a pro-drug to said subject, wherein the prodrug is converted to a cytotoxic compound by the action of the protein encoded by said cytotoxic gene and thereby killing the tumor cell expressing PDX-1.    
     
     
         137 . The method of  claim 136 , where the cytotoxic gene is the thymidine kinase gene.  
     
     
         138 . The method of  claim 136 , where the cytotoxic gene is the thymidine kinase gene and the prodrug is acyclovir, ganciclovir, FIAU or 6-methoxypurine arabinoside.  
     
     
         139 . The method of  claim 136 , wherein the administration is systemic.