Methods and compositions for the treatment of human and animal cancers

Abstract

Methods and compositions for altering the viability of cells, particularly cancers in animals and humans are disclosed. The compositions of the present invention are formed from a set of components comprising one or more of the following: a dithiocarbonyl, preferably dithiocarbamate, compound; a divalent metal ion; a modulator of cellular glutathione levels; and an inhibitor of the phosphorylation of choline. The compositions described herein induce a relatively selective and rapid effect on the viability of cancer cells by inducing a mixture of apoptotic and necrotic cell death, with the dominant pathway being apoptosis. Particularly preferred active compositions comprise all four components, although combinations of fewer components can be fully effective in certain tumors.

1 .- 10 . (Cancelled)  
     
     
         11 . A method for treatment of cancer cells, comprising the steps of: 
 providing a composition comprising 
 a biologically effective amount of a dithiocarbamate compound; and  
 a biologically effective amount of a modulator of cellular glutathione effective to decrease cellular glutathione levels, wherein the modulator of cellular glutathione is selected from the group consisting of ethacrynic acid, L-buthionine-S,R-sulfoximine, diethylmaleate, 2-cyclohexene-1-one, and 1-chloro-2,4-dinitrobenzene; and  
   causing the composition to contact the cancer cells.    
     
     
         12 . The method of  claim 11 , wherein the step of causing the composition to contact the cancer cells is effected by systemic, parenteral, intravenous, topical, oral, intratumoral, subcutaneous, intradermal, or transdermal administration of the composition.  
     
     
         13 . The method of  claim 11 , wherein the step of causing the composition to contact the cancer cells is effected by intratumoral administration.  
     
     
         14 .- 19 . (Cancelled)  
     
     
         20 . A method of identifying a therapeutic composition for treating one or more cancers, comprising the steps of: 
 preparing a plurality of test compositions, each test composition independently comprising at least two of the following components: 
 a dithiocarbamate compound;  
 a divalent metal ion;  
 a modulator of cellular glutathione; and  
 an inhibitor of the phosphorylation of choline;  
   examining the test compositions to determine if any produce results indicative of biological effect against one or more cancer cell types; and    selecting a therapeutic composition to be used in treating one or more cancers based on information comprising the results.    
     
     
         21 . The method of  claim 20 , wherein the biological effect is determined by measuring apoptosis, necrosis, DNA or cellular proliferation rates, or morphological cellular changes.  
     
     
         22 . The method of  claim 20 , wherein the dithiocarbamate compound is selected from the group consisting of diethyldithiocarbamate; tetraethylthiuram disulfide; and pyrrolidinedithiocarbamate.  
     
     
         23 . The method of  claim 20 , wherein the divalent metal ion is selected from the group consisting of Zn ++  and Cu ++ .  
     
     
         24 . The method of  claim 20 , wherein the modulator of cellular glutathione is selected from the group consisting of ethacrynic acid, L-buthionine-S,R-sulfoximine, diethylmaleate, 2-cyclohexene-1-one, and 1-chloro-2,4-dinitrobenzene.  
     
     
         25 . The method of  claim 20 , wherein the inhibitor of the phosphorylation of choline is dimethylethanolamine.  
     
     
         26 .- 30 . (Cancelled)  
     
     
         31 . The method of  claim 11 , wherein the method is used in combination with radiotherapy.  
     
     
         32 . The method of  claim 11 , wherein the method is used prior to surgical removal of the tumor.  
     
     
         33 . The method of  claim 32 , wherein the step of causing the composition to contact the cancer cells is effected by intratumoral administration.  
     
     
         34 . The method of  claim 33 , wherein the intratumoral administration occurs about 4 hours prior to surgical removal of the tumor.  
     
     
         35 . The method of  claim 33 , wherein the intratumoral administration occurs about 2 hours prior to surgical removal of the tumor.  
     
     
         36 . The method of  claim 11 , wherein the method is used to treat cancer cells in a surgical site remaining after surgical treatment of a primary tumor.  
     
     
         37 . The method of  claim 11 , wherein the dithiocarbamate compound has the formula: 
       (R 1 )(R 2 )N—C(═S)—S—Y, wherein R 1  and R 2  may be independently selected from the group consisting of hydrogen, C1-C24 straight, branched, or cyclic alkyl, alkenyl, aryl, acyl, alkaryl, aralkyl, and alkoxy groups, optionally substituted with ester, ether, halogen, sulfate, hydroxy, or phosphate groups, and wherein R 1  and R 2  may be optionally connected via a bridge comprising —(CH 2 ) n —, wherein n is 3-8, and wherein said bridge may be optionally substituted independently on any of the carbon atoms with C 1 -C 10  straight, branched, or cyclic alkyl, aryl, aryalkyl, or alkaryl groups, each of said groups optionally substituted with hydroxy, halo, phosphate, sulfate, or sulfonate groups; and    wherein Y is chosen from the group consisting of hydrogen, a pharmaceutically acceptable cation, a physiologically cleavable leaving group, a targeting moiety, and a chemotherapeutic drug.    
     
     
         38 . The method of  claim 11 , wherein the dithiocarbamate compound is selected from the group consisting of: diethyldithiocarbamate; tetraethylthiuram disulfide; and pyrrolidinedithiocarbamate.  
     
     
         39 . The method of  claim 11 , wherein the dithiocarbamate compound is pyrrolidinedithiocarbamate.  
     
     
         40 . The method of  claim 11 , wherein the modulator of cellular glutathione is ethacrynic acid.  
     
     
         41 . The method of  claim 11 , wherein the dithiocarbamate compound is pyrrolidinedithiocarbamate, and the modulator of cellular glutathione is ethacrynic acid.  
     
     
         42 . The method of  claim 41 , wherein the composition comprises about 10 to about 50 μM pyrrolidinedithiocarbamate, and about 10 to about 50 μM ethacrynic acid.  
     
     
         43 . The method of  claim 41 , wherein the composition comprises about 20 μM pyrrolidinedithiocarbamate, and about 10 μM ethacrynic acid.  
     
     
         44 . The method of  claim 11 , further comprising a biologically effective amount of dimethylethanolamine.  
     
     
         45 . The method of  claim 11 , wherein the composition further comprises a biologically effective amount of a divalent metal ion selected from the group consisting of Zn ++  and Cu ++ .  
     
     
         46 . The method of  claim 45 , wherein the divalent metal ion is Zn ++ .  
     
     
         47 . The method of  claim 45 , wherein the composition comprises about 5 to about 50 μM pyrrolidinedithiocarbamate, about 50 to about 200 μM Zn ++ , and about 10 to about 100 μM ethacrynic acid.  
     
     
         48 . The method of  claim 45 , wherein the composition comprises about 10 to about 50 μM pyrrolidinedithiocarbamate, about 30 to about 80 μM Zn ++ , and about 30 to about 80 μM ethacrynic acid.  
     
     
         49 . The method of  claim 45 , further comprising a biologically effective amount of dimethylethanolamine.  
     
     
         50 . The method of  claim 49 , wherein: 
 the dithiocarbamate compound is pyrrolidinedithiocarbamate;    the modulator of cellular glutathione is ethacrynic acid; and    the divalent metal ion is Zn ++ .    
     
     
         51 . The method of  claim 45 , wherein the step of causing the composition to contact the cancer cells is effected by systemic, parenteral, intravenous, topical, oral, intratumoral, subcutaneous, intradermal, or transdermal administration of the composition.  
     
     
         52 . The method of  claim 45 , wherein the step of causing the composition to contact the cancer cells is effected by intratumoral administration.  
     
     
         53 . The method of  claim 45 , wherein the method is used in combination with radiotherapy.  
     
     
         54 . The method of  claim 45 , wherein the method is used prior to surgical removal of the tumor.  
     
     
         55 . The method of  claim 54 , wherein the step of causing the composition to contact the cancer cells is effected by intratumoral administration.  
     
     
         56 . The method of  claim 55 , wherein the intratumoral administration occurs about 4 hours prior to surgical removal of the tumor.  
     
     
         57 . The method of  claim 55 , wherein the intratumoral administration occurs about 2 hours prior to surgical removal of the tumor.  
     
     
         58 . The method of  claim 45 , wherein the method is used to treat cancer cells in a surgical site remaining after surgical treatment of a primary tumor.  
     
     
         59 . The method of  claim 22 , wherein the test compositions comprise about 5 to 250 μM of the dithiocarbamate compound.  
     
     
         60 . The method of  claim 23 , wherein the test compositions comprise about 20 to 500 μM of the divalent metal ion.  
     
     
         61 . The method of  claim 24 , wherein the test compositions comprise about 10 to 300 μM of the modulator of cellular glutathione.  
     
     
         62 . The method of  claim 25 , wherein the test compositions comprise about 3 to 40 mM of dimethylethanolamine.  
     
     
         63 . A method for treatment of cancer cells, comprising the steps of: 
 providing a composition comprising 
 a biologically effective amount of a dithiocarbamate compound;  
 a biologically effective amount of a divalent metal ion selected from the group consisting of Zn ++  and Cu ++ ; and  
 a biologically effective amount of dimethylethanolamine; and  
   causing the composition to contact the cancer cells.    
     
     
         64 . The method of  claim 63 , wherein the dithiocarbamate compound has the formula: 
       (R 1 )(R 2 )N—C(═S)—S—Y, wherein R 1  and R 2  may be independently selected from the group consisting of hydrogen, C1-C24 straight, branched, or cyclic alkyl, alkenyl, aryl, acyl, alkaryl, aralkyl, and alkoxy groups, optionally substituted with ester, ether, halogen, sulfate, hydroxy, or phosphate groups, and wherein R 1  and R 2  may be optionally connected via a bridge comprising —(CH 2 ) n —, wherein n is 3-8, and wherein said bridge may be optionally substituted independently on any of the carbon atoms with C1-C10 straight, branched, or cyclic alkyl, aryl, aryalkyl, or alkaryl groups, each of said groups optionally substituted with hydroxy, halo, phosphate, sulfate, or sulfonate groups; and    wherein Y is chosen from the group consisting of hydrogen, a pharmaceutically acceptable cation, a physiologically cleavable leaving group, a targeting moiety, and a chemotherapeutic drug.    
     
     
         65 . The method of  claim 63 , wherein the dithiocarbamate compound is selected from the group consisting of: diethyldithiocarbamate; tetraethylthiuram disulfide; and pyrrolidinedithiocarbamate.  
     
     
         66 . The method of  claim 63 , wherein the dithiocarbamate compound is pyrrolidinedithiocarbamate.  
     
     
         67 . The method of  claim 63 , wherein the divalent metal ion is Zn ++ .  
     
     
         68 . A method for treatment of cancer cells, comprising the steps of: 
 providing a composition comprising 
 tricyclo-[5.2.1.O 2,6 ]-decyl-9[8]-xanthogenate; and  
 a modulator of cellular glutathione selected from the group consisting of ethacrynic acid, L-buthionine-S,R-sulfoximine, diethylmaleate, 2-cyclohexene-1-one, and 1-chloro-2,4-dinitrobenzene; and  
   causing the composition to contact cancer cells.    
     
     
         69 . The method of  claim 68 , wherein the modulator of cellular glutathione is ethacrynic acid.  
     
     
         70 . The method of  claim 68 , wherein the composition further comprises dimethylethanolamine.  
     
     
         71 . The method of  claim 68 , wherein the composition further comprises a divalent metal ion selected from the group consisting of Zn ++  and Cu ++ .  
     
     
         72 . The method of  claim 71 , wherein the divalent metal ion is Zn ++ .  
     
     
         73 . The method of  claim 68 , wherein the modulator of cellular glutathione is ethacrynic acid, and wherein the composition further comprises dimethylethanolamine and Zn ++ .  
     
     
         74 . The method of  claim 68 , wherein the step of causing the composition to contact the cancer cells is effected by systemic, parenteral, intravenous, topical, oral, intratumoral, subcutaneous, intradermal, or transdermal administration of the composition.  
     
     
         75 . The method of  claim 68 , wherein the step of causing the composition to contact the cancer cells is effected by intratumoral administration.  
     
     
         76 . The method of  claim 68 , wherein the method is used in combination with radiotherapy.  
     
     
         77 . The method of  claim 68 , wherein the method is used prior to surgical removal of the tumor.  
     
     
         78 . The method of  claim 77 , wherein the step of causing the composition to contact the cancer cells is effected by intratumoral administration.  
     
     
         79 . The method of  claim 78 , wherein the intratumoral administration occurs about 4 hours prior to surgical removal of the tumor.  
     
     
         80 . The method of  claim 78 , wherein the intratumoral administration occurs about 2 hours prior to surgical removal of the tumor.  
     
     
         81 . The method of  claim 68 , wherein the method is used to treat cancer cells in a surgical site remaining after surgical treatment of a primary tumor.