Methods and Compounds Useful to Induce Apoptosis in Cancer Cells

Abstract

The present invention provides a method for treating cancer in a mammal comprising contacting the cancer cells with a compound which is a apogossypol, derivative.

1 . A method of treating cancer in a subject, comprising administering to the subject a chemosensitizing agent selected from the group consisting of gossypol, apogossypol, derivatives of apogossypol, theaflavin, theaflavin-3′-gallate, theaflavanin, (−) gallocatechin-3-gallate (GCG), (−) epigallocatechin-3-gallate (EGCG), (−) catechin-3-gallate (CG), (−) epicatechin-3-gallate (ECG), derivatives of purpurogallin, and mixtures thereof, in combination with an anticancer agent. 
     
     
         2 . The method of  claim 1  wherein the derivative of aposossypol is a compound having formula (I): 
       
         
           
           
               
               
           
         
         wherein: each R 6 , R 8 , R 9  and R 10  is independently hydrogen, hydroxyl, —(C 1 -C 6 )alkyl, —O(C 1 -C 6 )alkyl, —(C 1 -C 6 )alkylhalo, —OC(O)(C 1 -C 6 )alkyl, or halo; each R 7  is independently hydrogen, —(C 3 -C 8 )cycloalkyl, —(C 6 -C 10 )aryl, or —(C 1 -C 6 )alkyl(C 6 -C 10 )aryl; or a pharmaceutically acceptable salt thereof. 
       
     
     
         3 . The method of  claim 2  wherein each R 6 , R 8 , R 9 , and R 10  group is independently hydrogen, —OH, —OCH 3 , —CF 3 , —CH 3 , —OC 2 H 5 , —OC(O)CH 3 , F, Cl, or Br. 
     
     
         4 . The method of  claim 2  wherein each R 7  group is independently hydrogen, —C 2 H 5 ; -i-Pr, n-Pr, n-Bu, t-Bu, i-Bu, s-Bu, or cyclohexyl. 
     
     
         5 . The method of  claim 2  wherein each R 6 , R 8 , and R 9 , is —OC(O)CH 3 ; each R 7  is i-propyl; and each R 10  is —CH 3 . 
     
     
         6 . The method of  claim 1  wherein the chemosensitizing agent and the anticancer agent are administered at the same time. 
     
     
         7 . The method of  claim 1  wherein the chemosensitizing agent is administered prior to the anticancer agent. 
     
     
         8 . The method of  claim 1  wherein the anticancer agent is Flavopiridol, Adriamycin, Etoposide, Taxol, cisplatin or a combination thereof. 
     
     
         9 . The method of  claim 1  wherein the purpurogallin derivative is 5D1, 1163, or 1142. 
     
     
         10 . The method of  claim 1  wherein the cancer is lung cancer, breast cancer, prostate cancer, colorectal cancer, or leukemia. 
     
     
         11 . The method of  claim 10  wherein the leukemia is acute lymphocytic leukemia, chronic lymphocytic leukemia, acute myelogenous leukemia, or chronic myelogenous leukemia. 
     
     
         12 . A method for inducing apoptosis, modulating caspase activity, or inducing cell death in a mammal comprising contacting target cells with a compound having the formula (I) 
       
         
           
           
               
               
           
         
       
       wherein: each R 6 , R 8 , R 9 , and R 10  are independently hydrogen, hydroxyl, —(C 1 -C 6 )alkyl, —O(C 1 -C 6 )alkyl, —(C 1 -C 6 )alkylhalo, —OC(O)(C 1 -C 6 )alkyl, or halo; each R 7  is independently hydrogen, —(C 3 -C 8 )cycloalkyl, —(C 6 -C 10 )aryl, or —(C 1 -C 6 )alkyl(C 6 -C 10 )aryl; or a pharmaceutically acceptable salt thereof, effective to induce apoptosis, modulate caspase activity, or induce cell death the target cells. 
     
     
         13 . The method of  claim 12  wherein each R 6 , R 8 , R 9 , and R 10  group is independently hydrogen, —OH, —OCH 3 , —CF 3 , —CH 3 , —OC 2 H 5 , —OC(O)CH 3 , F, Cl, or Br. 
     
     
         14 . The method of  claim 12  wherein each R 7  group is independently hydrogen, —C 2 H 5 ; -i-Pr, n-Pr, n-Bu, t-Bu, i-Bu, s-Bu, or cyclohexyl. 
     
     
         15 . The method of  claim 12  wherein each R 6 , R 8 , and R 9 , is —OC(O)CH 3 ; each R 7  is i-propyl; and each R 10  is —CH 3 . 
     
     
         16 . A method of identifying an agent that inhibits the anti-apoptotic activity of the Bcl-2 family proteins comprising:
 (a) identifying a Bcl-2 inhibitor or a labeled Bcl-2 inhibitor, wherein the inhibitor is selected from the group consisting of gossypol, apogossypol, derivatives of apogossypol, theaflavin, theaflavin-3′-gallate, theaflavanin, (−) gallocatechin-3-gallate (GCG), (−) epigallocatechin-3-gallate (EGCG), (−) catechin-3-gallate (CG), (−) epicatechin-3-gallate (ECG), and derivatives of purpurogallin;   (b) contacting the bound inhibitor with a candidate agent, said candidate agent suspected of being able to inhibit the Bcl-2 family proteins; and   (c) detecting dissociation of the inhibitor from the Bcl-2 family protein, whereby the agent is identified as an agent that inhibits Bcl-2 family proteins.   
     
     
         17 . The method of  claim 16  wherein the Bcl-2 family protein is Bcl-x L , Bcl-2, Mcl-1, Bcl-W, or Bcl-B. 
     
     
         18 . The method of  claim 16  wherein the inhibitor is labeled. 
     
     
         19 . The method of  claim 17  wherein the label is fluorescein, a fluorescein derivative, coumarin or a coumarin derivative. 
     
     
         20 . The method of  claim 19  wherein the label is fluorescein. 
     
     
         21 . The method of  claim 18  wherein the compound is labeled with an isotope. 
     
     
         22 . The method of  claim 21  wherein the isotope, is  13 C,  15 N,  19 F, or  1 H. 
     
     
         23 . The method of  claim 16  wherein the spectral technique comprises Nuclear Magnetic Resonance (NMR) binding assays. 
     
     
         24 . The method of  claim 16  wherein the spectral technique comprises, Fluorescence Polarization Assay (FPA).