US2003207341A1PendingUtilityA1

Methods for determining compounds capable of inhibiting the activities of the Ha-ras oncogene

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Assignee: UNIV COLUMBIAPriority: Nov 24, 1995Filed: Mar 24, 2003Published: Nov 6, 2003
Est. expiryNov 24, 2015(expired)· nominal 20-yr term from priority
Inventors:Paul FisherZao-Zhong Su
A61P 35/00A61K 38/00C07K 14/4703C07K 14/82
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Claims

Abstract

This invention provides a method for determining whether a compound is capable of suppressing ras functions comprising: (a) contacting an effective amount of the compound with Ha-ras transformed cloned rat embryo fibroblast cells under conditions permitting the compound to suppress ras functions in the cells; and (b) determining the expression or inhibition of certain indicator gene or genes, thereby determining whether the compound is capable of suppressing ras function. This invention further provides the determined compound and a pharmaceutical composition comprising the compound and a pharmaceutically acceptable carrier. This invention also provides methods for generating transcriptional switched Ha-ras transformed cloned rat embryo fibroblast cells. This invention also provides the generated cells and different uses of the cells.

Claims

exact text as granted — not AI-modified
What is claimed is:  
     
         1 . A method for determining whether a compound is capable of suppressing the ras functions comprising: 
 (a) contacting an effective amount of the compound with Ha-ras transformed cloned rat embryo fibroblast cells under conditions permitting the compound to suppress the ras functions in the cells; and    (b) determining the expression of lysyl oxidase, the expression of lysyl oxidase indicating that the compound is capable of suppressing the ras functions.    
     
     
         2 . A method for determining whether a compound is capable of suppressing the ras functions comprising: 
 (a) contacting an effective amount of the compound with Ha-ras transformed cloned rat embryo fibroblast cells under conditions permitting the compound to suppress the ras functions in the cells; and    (b) determining the expression of TIMP-1, the expression of TIMP-1 indicating that the compound is capable of suppressing the ras functions.    
     
     
         3 . A method for determining whether a compound is capable of suppressing the ras functions comprising: 
 (a) contacting an effective amount of the compound with Ha-ras transformed cloned rat embryo fibroblast cells under conditions permitting the compound to suppress the ras functions in the cells; and    (b) determining the expression of 92-kDa gelatinase, the inhibition of the expression of 92-kDa gelatinase indicating that the compound is capable of suppressing the ras functions.    
     
     
         4 . A method for determining whether a compound is capable of suppressing the ras functions comprising: 
 (a) contacting an effective amount of the compound with Ha-ras transformed cloned rat embryo fibroblast cells under conditions permitting the compound to suppress the ras functions in the cells; and    (b) determining the expression of transin, the inhibition of the expression of transin indicating that the compound is capable of suppressing the ras functions.    
     
     
         5 . A method of  claim 1 ,  2 ,  3  or  4 , where the compound was not previously known.  
     
     
         6 . The compound determined to be capable of suppressing the ras functions by method of  claim 5 .  
     
     
         7 . A pharmaceutical composition comprising the compound determined to be capable of suppressing the ras functions by method of  claim 1 ,  2 ,  3 , or  4  and a pharmaceutically acceptable carrier.  
     
     
         8 . A method for generating transcriptional switched Ha-ras transformed cloned rat embryo fibroblast cells comprising: 
 (a) introducing a gene to the Ha-ras transformed cloned rat embryo fibroblast cells; and    (b) determining the expression of lysyl oxidase in the introduced cells, the expression of lysyl oxidase indicating that the cells are transcriptional switched.    
     
     
         9 . A method for generating transcriptional switched Ha-ras transformed cloned rat embryo fibroblast cells comprising: 
 (a) introducing a gene to the Ha-ras transformed cloned rat embryo fibroblast cells; and    (b) determining the expression of TIMP-1 in the introduced cells, the expression of TIMP-1 indicating that the cells are transcriptional switched.    
     
     
         10 . A method for generating transcriptional switched Ha-ras transformed cloned rat embryo fibroblast cells comprising: 
 (a) introducing a gene to the Ha-ras transformed cloned rat embryo fibroblast cells; and    (b) determining expression of the 92-kDa gelatinase in the introduced cells, the inhibition of the expression of the 92-kDa gelatinase indicating that the cells are transcriptional switched.    
     
     
         11 . A method for generating transcriptional switched Ha-ras transformed cloned rat embryo fibroblast cells comprising: 
 (a) introducing a gene to the Ha-ras transformed cloned rat embryo fibroblast cells; and    (b) determining expression of the transin in the introduced cells, the inhibition of the expression of the transin indicating that the cells are transcriptional switched.    
     
     
         12 . A method of  claim 8 ,  9 ,  10 , or  11 , wherein the gene is a tumor suppressor gene.  
     
     
         13 . A method of  claim 12 , wherein the tumor suppressor gene is Krev-1.  
     
     
         14 . Cells generated by methods of  claim 8 ,  9 ,  10 ,  11 ,  12  or  13 .  
     
     
         15 . A method to isolate genes which are associated with transcriptional switching of the Ha-ras transformed cloned rat embryo fibroblast cells comprising: 
 (a) selecting genes which are either expressed or suppressed in the transcriptional switched Ha-ras transformed cloned rat embryo fibroblast cells; and    (b) isolating the selected gene.    
     
     
         16 . A method of  claim 15 , wherein the genes are selected by subtractive hybridization.  
     
     
         17 . A method of  claim 15 , wherein the genes are selected by differential display.  
     
     
         18 . Genes isolated by the method of  claim 15 ,  16  or  17 .  
     
     
         19 . Polypeptides encoded by the genes of  claim 18 .  
     
     
         20 . Molecules capable of competitively inhibiting the functions of the polypeptides of  claim 19 .  
     
     
         21 . Molecules capable of mimicking the functions of the polypeptides of  claim 19.

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