Method of modulating ribonucleotide reductase

Abstract

A method of modulating ribonucleotide reductase activity in a neoplastic cell includes administering to the cell an amount of a ribonucleotide reductase allosteric modulator (RRAmod), the amount being effective to inhibit neoplastic cell growth.

Having described the invention, the following is claimed: 
     
         1 . A method of modulating ribonucleotide reductase activity in a neoplastic cell comprising administering to the cell an amount of a ribonucleotide reductase allosteric modulator (RRAmod), the amount being the amount effective to inhibit neoplastic cell growth. 
     
     
         2 . The method of  claim 1 , the RRAmod modulating ribonucleotide reductase activity by selectively binding at the hexamer interface of RR1. 
     
     
         3 . The method of  claim 2 , the hexamer interface of RR1 comprising an epitope having an amino acid sequence corresponding to SEQ ID NO: 1. 
     
     
         4 . The method of  claim 1 , wherein modulating ribonucleotide reductase activity comprises modulating ribonucleotide reductase mediated catalyzation of ribonucleotides to deoxy ribonucleotides in the neoplastic cell, thereby unbalancing the nucleotide pool of DNA precursor molecules required for de novo DNA synthesis. 
     
     
         5 . The method of  claim 1 , the RRAmod comprising a small molecule, a peptide, a peptidomimetic, or an antibody. 
     
     
         6 . The method of  claim 1 , the RRAmod comprising a small molecule having the formula: 
       
         
           
           
               
               
           
         
         wherein each of R 1  to R 20 , R 46  to R 63  can be independently selected from the group consisting of hydrogen, C 1 -C 24  alkyl, C 2 -C 24  alkenyl, C 2 -C 24  alkynyl, C 3 -C 20  aryl, C 6 -C 24  alkaryl, C 6 -C 24  aralkyl, halo, silyl, hydroxyl, sulfhydryl, C 1 -C 24  alkoxy, C 2 -C 24  alkenyloxy, C 2 -C 24  alkynyloxy, C 5 -C 20  aryloxy, acyl (including C 2 -C 24  alkylcarbonyl (—CO-alkyl) and C 6 -C 20  arylcarbonyl (—CO-aryl)), acyloxy (—O-acyl), C 2 -C 24  alkoxycarbonyl (—(CO)—O-alkyl), C 6 -C 20  aryloxycarbonyl (—(CO)—O-aryl), C 2 -C 24  alkylcarbonato (—O—(CO)—O-alkyl), C 6 -C 20  arylcarbonato (—O—(CO)—O-aryl), carboxy (—COOH), carboxylato (—COO − ), carbamoyl (—(CO)—NH 2 ), C 1 -C 24  alkyl-carbamoyl (—(CO)—NH(C 1 -C 24  alkyl)), arylcarbamoyl (—(CO)—NH-aryl), thiocarbamoyl (—(CS)—NH 2 ), carbamide (—NH—(CO)—NH 2 ), cyano(—CN), isocyano (—N + C − ), cyanato (—O—CN), isocyanato (—O—N + ═C − ), isothiocyanato (—S—CN), azido (—N═N + ═N − ), formyl (—(CO)—H), thioformyl (—(CS)—H), amino (—NH 2 ), C 1 -C 24  alkyl amino, C 5 -C 20  aryl amino, C 2 -C 24  alkylamido (—NH—(CO)-alkyl), C 6 -C 20  arylamido (—NH—(CO)-aryl), imino (—CR═NH where R is hydrogen, C 1 -C 24  alkyl, C 5 -C 20  aryl, C 6 -C 24  alkaryl, C 6 -C 24  aralkyl, etc.), alkylimino (—CR═N(alkyl), where R=hydrogen, alkyl, aryl, alkaryl, aralkyl, etc.), arylimino (—CR═N(aryl), where R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro (—NO 2 ), nitroso (—NO), sulfo (—SO 2 —OH), sulfonato (—SO 2 —O − ), C 1 -C 24  alkylsulfanyl (—S-alkyl; also termed “alkylthio”), arylsulfanyl (—S-aryl; also termed “arylthio”), C 1 -C 24  alkylsulfinyl (—(SO)-alkyl), C 5 -C 20  arylsulfinyl (—(SO)-aryl), C 1 -C 24  alkylsulfonyl (—SO 2 -alkyl), C 5 -C 20  arylsulfonyl (—SO 2 -aryl), phosphono (—P(O)(OH) 2 ), phosphonato (—P(O)(O − ) 2 ), phosphinato (—P(O)(O − )), phospho (—PO 2 ), phosphino (—PH 2 ), combinations thereof, and wherein adjacent R groups may be linked to form a cyclic or polycyclic ring, wherein the cyclic ring is aromatic, alicyclic, heteroaromatic, or heterocyclic; 
         X 1  is a C 1 -C 24  alkyl, C 2 -C 24  alkenyl, C 2 -C 24  alkynyl; 
         Y 1  is a C 1 -C 24  alkyl, C 2 -C 24  alkenyl, C 2 -C 24  alkynyl; 
         R 45  is hydrogen, a C 1 -C 24  alkyl, C 2 -C 24  alkenyl, C 2 -C 24  alkynyl, an amino (—NH 2 ), or —(C 1 -C 24  alkyl)-amino, and combinations thereof; 
         and a pharmaceutically acceptable salt thereof. 
       
     
     
         7 . The method of  claim 6 , wherein the RRAmod is a small molecule selected from the group consisting of: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         and pharmaceutically acceptable salts thereof. 
       
     
     
         8 . The method of  claim 1 , the cell comprising a cancer cell. 
     
     
         9 . The method of  claim 8 , the cancer cell comprising a pancreatic, breast, lung, colon or glyoblastoma cancer cell. 
     
     
         10 . A method of treating a neoplastic disorder in a subject comprising:
 administering to neoplastic cells of the subject a therapeutically effective amount of a pharmaceutical composition, the composition comprising a ribonucleotide reductase allosteric modulator (RRAmod), the therapeutically effective amount being the amount to inhibit neoplastic cell growth in the subject.   
     
     
         11 . The method of  claim 10 , the RRAmod modulating ribonucleotide reductase activity by selectively binding to the hexamer interface of RR1. 
     
     
         12 . The method of  claim 11 , the hexamer interface of RR1 comprising an epitope having an amino acid sequence corresponding to SEQ ID NO: 1. 
     
     
         13 . The method of  claim 10 , wherein modulating ribonucleotide reductase activity comprises modulating ribonucleotide reductase mediated catalyzation of ribonucleotides to deoxy ribonucleotides in the cell, thereby unbalancing the nucleotide pool of DNA precursor molecules required for de novo DNA synthesis. 
     
     
         14 . The method of  claim 10 , the RRAmod comprising a small molecule having the formula: 
       
         
           
           
               
               
           
         
         wherein each of R 1  to R 20 , R 46  to R 63  can be independently selected from the group consisting of hydrogen, C 1 -C 24  alkyl, C 2 -C 24  alkenyl, C 2 -C 24  alkynyl, C 3 -C 20  aryl, C 6 -C 24  alkaryl, C 6 -C 24  aralkyl, halo, silyl, hydroxyl, sulfhydryl, C 1 -C 24  alkoxy, C 2 -C 24  alkenyloxy, C 2 -C 24  alkynyloxy, C 5 -C 20  aryloxy, acyl (including C 2 -C 24  alkylcarbonyl (—CO-alkyl) and C 6 -C 20  arylcarbonyl (—CO-aryl)), acyloxy (—O-acyl), C 2 -C 24  alkoxycarbonyl (—(CO)—O-alkyl), C 6 -C 20  aryloxycarbonyl (—(CO)—O-aryl), C 2 -C 24  alkylcarbonato (—O—(CO)—O-alkyl), C 6 -C 20  arylcarbonato (—O—(CO)—O-aryl), carboxy (—COOH), carboxylato (—COO − ), carbamoyl (—(CO)—NH 2 ), C 1 -C 24  alkyl-carbamoyl (—(CO)—NH(C 1 -C 24  alkyl)), arylcarbamoyl (—(CO)—NH-aryl), thiocarbamoyl (—(CS)—NH 2 ), carbamide (—NH—(CO)—NH 2 ), cyano(—CN), isocyano (—N + C − ), cyanato (—O—CN), isocyanato (—O—N + ═C − ), isothiocyanato (—S—CN), azido (—N═N + ═N − ), formyl (—(CO)—H), thioformyl (—(CS)—H), amino (—NH 2 ), C 1 -C 24  alkyl amino, C 5 -C 20  aryl amino, C 2 -C 24  alkylamido (—NH—(CO)-alkyl), C 6 -C 20  arylamido (—NH—(CO)-aryl), imino (—CR═NH where R is hydrogen, C 1 -C 24  alkyl, C 5 -C 20  aryl, C 6 -C 24  alkaryl, C 6 -C 24  aralkyl, etc.), alkylimino (—CR═N(alkyl), where R=hydrogen, alkyl, aryl, alkaryl, aralkyl, etc.), arylimino (—CR═N(aryl), where R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro (—NO 2 ), nitroso (—NO), sulfo (—SO 2 —OH), sulfonato (—SO 2 —O − ), C 1 -C 24  alkylsulfanyl (—S-alkyl; also termed “alkylthio”), arylsulfanyl (—S-aryl; also termed “arylthio”), C 1 -C 24  alkylsulfinyl (—(SO)-alkyl), C 5 -C 20  arylsulfinyl (—(SO)-aryl), C 1 -C 24  alkylsulfonyl (—SO 2 -alkyl), C 5 -C 20  arylsulfonyl (—SO 2 -aryl), phosphono (—P(O)(OH) 2 ), phosphonato (—P(O)(O − ) 2 ), phosphinato (—P(O)(O − )), phospho (—PO 2 ), phosphino (—PH 2 ), combinations thereof, and wherein adjacent R groups may be linked to form a cyclic or polycyclic ring, wherein the cyclic ring is aromatic, alicyclic, heteroaromatic, or heterocyclic; 
         X 1  is a C 1 -C 24  alkyl, C 2 -C 24  alkenyl, C 2 -C 24  alkynyl; 
         Y 1  is a C 1 -C 24  alkyl, C 2 -C 24  alkenyl, C 2 -C 24  alkynyl; 
         R 45  is hydrogen, a C 1 -C 24  alkyl, C 2 -C 24  alkenyl, C 2 -C 24  alkynyl, an amino (—NH 2 ), or —(C 1 -C 24  alkyl)-amino, and combinations thereof; 
         and a pharmaceutically acceptable salt thereof. 
       
     
     
         15 . The method of  claim 14 , wherein the RRAmod is a small molecule selected from the group consisting of: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         and pharmaceutically acceptable salts thereof. 
       
     
     
         16 . The method of  claim 10 , the neoplastic disorder comprising cancer. 
     
     
         17 . The method of  claim 16 , wherein the cancer includes pancreatic, breast, lung, colon or glyoblastoma cancer. 
     
     
         18 . The method of  claim 10 , further administering another therapeutic agent in conjunction with the RRAmod. 
     
     
         19 . The method of  claim 18 , the other therapeutic agent comprising at least one of a chemotherapeutic agent, an antimetabolite, a DNA damaging agent, a ribonucleotide reductase inhibiting agent, an antitumorgenic agent, an antimitotic agent, an antiviral agent, an antineoplastic agent, an immunotherapeutic agent, and a radiotherapeutic agent. 
     
     
         20 . A pharmaceutical composition comprising a ribonucleotide reductase allosteric modulator (RRAmod), the RRAmod inhibiting cell growth when administered to a neoplastic cell, wherein the RRAmod includes a small molecule having the formula: 
       
         
           
           
               
               
           
         
         wherein each of R 1  to R 20 , R 46  to R 63  can be independently selected from the group consisting of hydrogen, C 1 -C 24  alkyl, C 2 -C 24  alkenyl, C 2 -C 24  alkynyl, C 3 -C 20  aryl, C 6 -C 24  alkaryl, C 6 -C 24  aralkyl, halo, silyl, hydroxyl, sulfhydryl, C 1 -C 24  alkoxy, C 2 -C 24  alkenyloxy, C 2 -C 24  alkynyloxy, C 5 -C 20  aryloxy, acyl (including C 2 -C 24  alkylcarbonyl (—CO-alkyl) and C 6 -C 20  arylcarbonyl (—CO-aryl)), acyloxy (—O-acyl), C 2 -C 24  alkoxycarbonyl (—(CO)—O-alkyl), C 6 -C 20  aryloxycarbonyl (—(CO)—O-aryl), C 2 -C 24  alkylcarbonato (—O—(CO)—O-alkyl), C 6 -C 20  arylcarbonato (—O—(CO)—O-aryl), carboxy (—COOH), carboxylato (—COO − ), carbamoyl (—(CO)—NH 2 ), C 1 -C 24  alkyl-carbamoyl (—(CO)—NH(C 1 -C 24  alkyl)), arylcarbamoyl (—(CO)—NH-aryl), thiocarbamoyl (—(CS)—NH 2 ), carbamide (—NH—(CO)—NH 2 ), cyano(—CN), isocyano (—N + C − ), cyanato (—O—CN), isocyanato (—O—N + ═C − ), isothiocyanato (—S—CN), azido (—N═N + ═N − ), formyl (—(CO)—H), thioformyl (—(CS)—H), amino (—NH 2 ), C 1 -C 24  alkyl amino, C 5 -C 20  aryl amino, C 2 -C 24  alkylamido (—NH—(CO)-alkyl), C 6 -C 20  arylamido (—NH—(CO)-aryl), imino (—CR═NH where R is hydrogen, C 1 -C 24  alkyl, C 5 -C 20  aryl, C 6 -C 24  alkaryl, C 6 -C 24  aralkyl, etc.), alkylimino (—CR═N(alkyl), where R=hydrogen, alkyl, aryl, alkaryl, aralkyl, etc.), arylimino (—CR═N(aryl), where R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro (—NO 2 ), nitroso (—NO), sulfo (—SO 2 —OH), sulfonato (—SO 2 —O − ), C 1 -C 24  alkylsulfanyl (—S-alkyl; also termed “alkylthio”), arylsulfanyl (—S-aryl; also termed “arylthio”), C 1 -C 24  alkylsulfinyl (—(SO)-alkyl), C 5 -C 20  arylsulfinyl (—(SO)-aryl), C 1 -C 24  alkylsulfonyl (—SO 2 -alkyl), C 5 -C 20  arylsulfonyl (—SO 2 -aryl), phosphono (—P(O)(OH) 2 ), phosphonato (—P(O)(O − ) 2 ), phosphinato (—P(O)(O − )), phospho (—PO 2 ), phosphino (—PH 2 ), combinations thereof, and wherein adjacent R groups may be linked to form a cyclic or polycyclic ring, wherein the cyclic ring is aromatic, alicyclic, heteroaromatic, or heterocyclic; 
         X 1  is a C 1 -C 24  alkyl, C 2 -C 24  alkenyl, C 2 -C 24  alkynyl; 
         Y 1  is a C 1 -C 24  alkyl, C 2 -C 24  alkenyl, C 2 -C 24  alkynyl; 
         R 45  is hydrogen, a C 1 -C 24  alkyl, C 2 -C 24  alkenyl, C 2 -C 24  alkynyl, an amino (—NH 2 ), —(C 1 -C 24  alkyl)-amino, and combinations thereof; 
         and a pharmaceutically acceptable salt thereof. 
       
     
     
         21 . The pharmaceutical composition of  claim 20 , wherein the RRAmod is a small molecule selected from the group consisting of: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         and pharmaceutically acceptable salts thereof. 
       
     
     
         22 . The pharmaceutical composition of  claim 20 , the neoplastic cell comprising a cancer cell. 
     
     
         23 . The pharmaceutical composition of  claim 24 , the cancer cell comprising a pancreatic, breast, lung, colon or glyoblastoma cancer cell. 
     
     
         24 . The pharmaceutical composition of  claim 20 , further comprising another therapeutic agent in conjunction with the RRAmod. 
     
     
         25 . The pharmaceutical composition of  claim 24 , the other therapeutic agent comprising at least one of a chemotherapeutic agent, an antimetabolite, a DNA damaging agent, a ribonucleotide reductase inhibiting agent, an antitumorgenic agent, an antimitotic agent, an antiviral agent, an antineoplastic agent, an immunotherapeutic agent, and a radiotherapeutic agent.