Fluorocyclopentenylcytosine methods of use

Abstract

or a hydrate, a solvate, or a pharmaceutically acceptable salt thereof. The disclosed subject matter further provides a method of treating one or more symptoms of cancer comprising administering to a subject in need thereof a compound of formula (I) and a process for preparing such.

1 . A process for preparing of 4-amino-1-((1S,4R,5S)-2-fluoro-4,5-dihydroxy-3-(hydroxymethyl)cyclopent-2-en-1-yl)pyrimidin-2(1H)-one 1H 2 O (RX-3117-MH), comprising converting tert-butyl(((3aR,4R,6aR)-5-fluoro-2,2-dimethyl-6-((trityloxy)methyl)-4,6a-dihydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)oxy)diphenylsilane (ASM11) to 4-amino-1-((3aS,4S,6aR)-5-fluoro-2,2-dimethyl-6-((trityloxy)methyl)-4,6a-dihydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)pyrimidin-2(1H)-one (INT14) in a continuous process with more than one step without isolation of any intermediate. 
     
     
         2 . The process of  claim 1 , further comprising the steps of:
 dissolving tert-butyl(((3aR,4R,6aR)-5-fluoro-2,2-dimethyl-6-((trityloxy)methyl)-4,6a-dihydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)oxy)diphenylsilane (ASM11) in 2-methyl tetrahydrofuran;   adding tetra-n-butylammonium fluoride to form ((3aS,4R,6aR)-5-fluoro-2,2-dimethyl-6-((trityloxy)methyl)-4,6a-dihydro-3aH-cyclopenta[d][1,3]dioxol-4-ol) (INT12) in a reaction solution; and   recovering INT12 in an organic phase.   
     
     
         3 . The process of  claim 2 , wherein said recovering INT12 in the organic phase comprises the steps of:
 washing the reaction solution with an aqueous solution;   separating an aqueous extraction from the organic phase having INT12;   washing the aqueous extraction with 2-methyl tetrahydrofuran to extract INT12 from the aqueous extraction; and   combining the extracted INT12 with the organic phase having INT12.   
     
     
         4 . The process of  claim 2 , further comprising:
 adding triethylamine and methanesulphonyl chloride in 2-methyl tetrahydrofuran to the INT12 in the organic phase to form ((3aR,4R,6aR)-5-fluoro-2,2-dimethyl-6-((trityloxy)methyl)-4,6a-dihydro-3aH-cyclopenta[d][1,3]dioxol-4-yl methanesulfonate) (INT13) in a second reaction solution; and   recovering INT13 in DMSO.   
     
     
         5 . The process of  claim 4 , wherein said recovering INT13 in DMSO comprises the steps of:
 adding the DMSO to the second reaction solution with INT13; and   removing at least 90% w/w of 2-methyl tetrahydrofuran by distillation.   
     
     
         6 . The process of  claim 4 , further comprising:
 adding 2.5 equivalents of cesium carbonate and cytosine to the INT13 in DMSO to form 4-amino-1-((3 aS,4 S,6aR)-5-fluoro-2,2-dimethyl-6-((trityloxy)methyl)-4,6a-dihydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)pyrimidin-2(1H)-one (INT14) in a third reaction solution.   
     
     
         7 . The process of  claim 6 , further comprising:
 maintaining reaction temperature at about 33 to 37° C.   
     
     
         8 . The process of  claim 6 , wherein the INT14 has a ratio of N- to O-isomers of over about 95:5. 
     
     
         9 . The process of  claim 6 , further comprising:
 adding an acid to the third reaction solution with INT14 to form 4-amino-1-((1S,4R,5S)-2-fluoro-4,5-dihydroxy-3-(hydroxymethyl)cyclopent-2-en-1-yl)pyrimidin-2(1H)-one (RX-3117);   washing the RX-3117 with methyl tert-butyl ether and water to form an organic phase and an aqueous phase having RX-3117; and   purifying the RX-3117 to form RX-3117-MH.   
     
     
         10 . The process of  claim 9 , further comprising:
 charging the reaction mixture with methanol and distilling the reaction mixture to remove acetonide until less than about 1.0% area of the acetonide is detected prior to the washing step.   
     
     
         11 . The process of  claim 9 , wherein the washing step further comprises:
 separating the aqueous phase having RX-3117 from the organic phase;   washing the aqueous phase having RX-3117 with methyl tert-butyl ether until less than about 0.5% w/w trityl alcohol is detected in the aqueous phase;   adding a basic anion resin to the aqueous phase having RX-3117 to form a slurry;   filtering the slurry to retain a mother liquor;   concentrating the mother liquor to form a concentrate; and   adding acetonitrile to the concentrate to form purified RX-3117-MH.   
     
     
         12 . A continuous process for preparing 4-amino-1-((3aS,4S,6aR)-5-fluoro-2,2-dimethyl-6-((trityloxy)methyl)-4,6a-dihydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)pyrimidin-2(1H)-one (INT14) from tert-butyl(((3aR,4R,6aR)-5-fluoro-2,2-dimethyl-6-((trityloxy)methyl)-4,6a-dihydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)oxy)diphenylsilane (ASM11), comprising the steps of:
 dissolving the ASM11 in 2-methyl tetrahydrofuran;   adding tetra-n-butylammonium fluoride to form ((3aS,4R,6aR)-5-fluoro-2,2-dimethyl-6-((trityloxy)methyl)-4,6a-dihydro-3aH-cyclopenta[d][1,3]dioxol-4-ol) (INT12);   adding trimethylamine and methanesulphonyl chloride in 2-methyl tetrahydrofuran to the INT12 to form ((3aR,4R,6aR)-5-fluoro-2,2-dimethyl-6-((trityloxy)methyl)-4,6a-dihydro-3aH-cyclopenta[d][1,3]dioxol-4-yl methanesulfonate) (INT13); and   adding cesium carbonate and cytosine to the INT13 to form 4-amino-1-((3aS,4S,6aR)-5-fluoro-2,2-dimethyl-6-((trityloxy)methyl)-4,6a-dihydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)pyrimidin-2(1H)-one (INT14);   wherein the steps are performed in one or more fixed reactors without isolation of INT12 or INT13.   
     
     
         13 . A continuous process for preparing 4-amino-1-((1S,4R,5S)-2-fluoro-4,5-dihydroxy-3-(hydroxymethyl)cyclopent-2-en-1-yl)pyrimidin-2(1H)-one 1H 2 O (RX-3117-MH) from 4-amino-1-((3 aS,4 S,6aR)-5-fluoro-2,2-dimethyl-6-((trityloxy)methyl)-4,6a-dihydro-3aH-cyclopenta[d][1,3]dioxol-4-yl)pyrimidin-2(1H)-one (INT14), comprising the steps of:
 reacting the INT14 with an acid to form 4-amino-1-((1 S,4R,5S)-2-fluoro-4,5-dihydroxy-3-(hydroxymethyl)cyclopent-2-en-1-yl)pyrimidin-2(1H)-one (RX-3117);   washing the RX-3117 with methyl tert-butyl ether and water to form an organic phase and an aqueous phase having RX-3117;   separating the aqueous phase having RX-3117 from the organic phase;   washing the aqueous phase having RX-3117 with methyl tert-butyl ether until less than about 0.5% w/w trityl alcohol is detected in the aqueous phase;   adding a strongly basic anion resin to the aqueous phase having RX-3117 to form a slurry;   filtering the slurry to retain a mother liquor;   concentrating the mother liquor to form a concentrate;   adding acetonitrile to the concentrate to form purified RX-3117-MH; and   isolating the purified RX-3117-MH;   wherein the steps are performed in one or more fixed reactors.   
     
     
         14 . A method of sensitizing a cell to an apoptotic signal comprising contacting the cell with an effective amount of a compound of formula (I) 
       
         
           
           
               
               
           
         
         or a hydrate, a solvate, or a pharmaceutically acceptable salt thereof. 
       
     
     
         15 . A method of treating one or more symptoms of cancer in a subject, comprising administering a compound of formula (I) 
       
         
           
           
               
               
           
         
         or a hydrate, a solvate, or a pharmaceutically acceptable salt thereof in an amount effective to inhibit methyltransferase and to upregulate at least one hypomethylated target in the subject.