Recombinant p53 adenovirus methods and compositions

Abstract

Described are simplified and efficient methods for preparing recombinant adenovirus using liposome-mediated cotransfection and the direct. observation of a cytopathic effect (CPE) in the transfected cells. Also disclosed are compositions and methods involving novel p53 adenovirus constructs, including methods for restoring p53 function and tumor suppression in cells and animals having abnormal p53.

1 - 21 . (canceled)  
     
     
         22 . A pharmaceutical composition comprising a pharmacologically acceptable solution or buffer and a replication-deficient adenovirus comprising a nucleic acid sequence encoding 
 i) wild-type human p53 under the control of a promoter; and    ii) a polyadenylation signal,    wherein the nucleic acid sequence replaces the E1 A and E1 B regions and the adenovirus comprises adenovirus serotype 5 sequence.    
     
     
         23 . The pharmaceutical composition of  claim 22 , wherein the adenovirus lacks E2, E3, or E4 region.  
     
     
         24 . The pharmaceutical composition of  claim 23 , wherein the adenovirus lacks E2, E3, and E4 regions.  
     
     
         25 . The pharmaceutical composition of  claim 22 , wherein the adenovirus also comprises an origin of replication.  
     
     
         26 . A method comprising administering to a cancer patient the pharmaceutical composition of  claim 22 .  
     
     
         27 . The method of  claim 26 , wherein the patient is administered a pharmaceutical composition containing about 10 3  to about 5×10 12  adenoviruses.  
     
     
         28 . The method of  claim 27 , wherein the patient is administered a pharmaceutical composition containing about 10 10  to about 5×10 12  adenoviruses.  
     
     
         29 . The method of  claim 26 , wherein the composition is injected or infused into the patient.  
     
     
         30 . The method of  claim 26 , wherein the cancer comprises malignant cells.  
     
     
         30 . The method of  claim 26 , wherein the composition is administered after resection of cancer cells.  
     
     
         31 . The method of  claim 26 , wherein the patient has an unresectable cancer.  
     
     
         32 . The method of  claim 26 , wherein the cancer is an epithelial cell cancer.  
     
     
         33 . The method of  claim 26 , wherein the cancer is lung cancer, breast cancer  
     
     
         34 . The method of  claim 26 , wherein the composition is a volume of 10 ml or less.  
     
     
         35 . The method of  claim 26 , wherein the patient receives at least two courses of therapy.