METHODS AND COMPOUNDS FOR RESTORING MUTANT p53 FUNCTION

Abstract

Mutations in oncogenes and tumor suppressors contribute to the development and progression of cancer. The present disclosure describes compounds and methods to recover wild-type function to p53 mutants. The compounds of the present invention can bind to mutant p53 and restore the ability of the p53 mutant to bind DNA and activate downstream effectors involved in tumor suppression. The disclosed compounds can be used to reduce the progression of cancers that contain a p53 mutation.

What is claimed is: 
     
         1 . A method of treating a condition, the method comprising administering to a subject in need thereof a therapeutically-effective amount of a compound that binds a p53 mutant in the subject, wherein the binding of the compound to the p53 mutant increases the ability of the p53 mutant to bind DNA by at least about 50% as compared to the ability of the p53 mutant to bind DNA in absence of the compound as determined by an assay, wherein the assay comprises:
 a) contacting the compound with a tagged mutant p53 moiety and an antibody conjugated to a fluorescence energy acceptor against a tag of the tagged mutant p53 moiety in a test chamber;   b) contacting the tagged mutant p53 moiety with a biotin-labeled DNA and streptavidin conjugated to a fluorescence energy donor in the test chamber;   c) irradiating the test chamber with light that promotes fluorescence resonance energy transfer;   d) detecting the fluorescence resonance energy transfer;   e) determining an SC 150  value of the compound based on the fluorescence resonance energy transfer;   f) comparing the SC 150  value of the compound with that of a control sample, wherein the control sample comprises the tagged mutant p53 moiety, the antibody conjugated to the fluorescence energy acceptor against the tag of the tagged mutant p53 moiety, the biotin-labeled DNA, and the streptavidin conjugated to the fluorescence energy donor, wherein the control sample does not comprise the compound; and   g) determining based on the comparing a level of activation of protein-DNA binding in the presence of the compound.   
     
     
         2 . The method of  claim 1 , wherein the p53 mutant has a mutation at amino acid 220. 
     
     
         3 . The method of  claim 2 , wherein the p53 mutant is p53 Y220C. 
     
     
         4 . The method of  claim 1 , wherein the compound induces a conformational change in the p53 mutant. 
     
     
         5 . The method of  claim 1 , wherein the compound selectively binds the p53 mutant as compared to wild type p53. 
     
     
         6 . The method of  claim 1 , wherein the binding of the compound to the p53 mutant induces apoptosis in a cell. 
     
     
         7 . The method of  claim 1 , wherein the binding of the compound to the p53 mutant induces cell cycle arrest in a cell. 
     
     
         8 . The method of  claim 1 , wherein the therapeutically-effective amount is from about 20 mg to about 2000 mg. 
     
     
         9 . The method of  claim 1 , wherein the condition is cancer. 
     
     
         10 . The method of  claim 1 , wherein the condition is ovarian cancer. 
     
     
         11 . The method of  claim 1 , wherein the condition is breast cancer. 
     
     
         12 . The method of  claim 1 , wherein the condition is lung cancer. 
     
     
         13 . The method of  claim 1 , wherein the administration is oral. 
     
     
         14 . The method of  claim 1 , wherein the compound increases the stability of a biologically-active conformation of the p53 mutant relative to the stability of the biologically-active conformation of the p53 mutant in absence of the compound. 
     
     
         15 . The method of  claim 1 , wherein the tagged p53 is His-tagged. 
     
     
         16 . The method of  claim 15 , wherein the tagged p53 comprises a Y220C mutation. 
     
     
         17 . The method of  claim 15 , wherein the tagged p53 comprises amino acids 94-312 of wild type p53. 
     
     
         18 . The method of  claim 1 , wherein the antibody is conjugated to allophycocyanin (APC). 
     
     
         19 . The method of  claim 1 , wherein the DNA is a DNA duplex of SEQ ID NO.: 2. 
     
     
         20 . The method of  claim 1 , wherein the FRET assay is a homogeneous time-resolved fluorescence (HRTF) assay.