Synthesis of fluorinated nucleotides
The present invention relates to efficient processes useful in the preparation of fluorinated nucleosides, such as (2S,3R,4S,5R)-5-(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)-3-fluoro-4-hydroxy-2-(mercaptomethyl)tetrahydrofuran-3-yl dihydrogen phosphate, also known as 3′-fluoro-thio-guanosine monophosphate or 3′-F-thio-GMP. Such fluorinated nucleosides may be useful as a biologically active compound and or as an intermediate for the synthesis of more complex biologically active compounds. The present invention also encompasses intermediates useful in the disclosed synthetic processes and the methods of their preparation. (I)
1 . A process for preparing a compound of Formula (I), or a pharmaceutically acceptable salt, hydrate, or solvate thereof:
wherein each R is independently selected from the group consisting of H, Na, and K, comprising
reacting a compound of Formula (I-2) with at least one Reducing Agent to provide a compound of Formula (I-1):
reacting the compound of Formula (I-1) with a thiophosphorylating agent in the presence of at least one Catalyst A and at least one Base A in the presence of at least one Solvent A, to form a compound of Formula (I-1′) and then reacting the compound of Formula (I-1′) with at least one Base B in the presence of at least one Solvent B:
wherein each R 1 is selected from the group consisting of C 1 -C 12 alkyl, benzyl, aryl, and heteroaryl, wherein
the at least one thiophosphorylating agent is thiophosphoryl chloride; and
the at least one Catalyst A is selected from the group consisting of
2 . The process according to claim 1 , wherein the compound of Formula (I) is a compound of Formula (Ia), or a pharmaceutically acceptable salt, hydrate, or solvate thereof:
3 . The process according to claim 1 , wherein the at least one Base A is selected from the group consisting of 2,6-lutidine, pyridine, Et 3 N, 1,8-bis(dimethylamino) naphthalene, 2,6-lutidine, 2,4-lutidine, 2-methyl-pyridine, trimethylpyridine, 3-methoxy-pyridine, 4-methyl-pyridine, quinuclidine, Hunig's base, 3-methyl-pyridine, and 2,6-di-tert-butyl-4-methyl pyridine, and mixtures thereof.
4 . The process according to claim 1 , wherein the at least one Solvent A is selected from the group consisting of THE, MeCN, acetone, DMPU, HFIP, TFE, DME, DMAC, 2-Me-THF, EtOAc, and MIBK, and mixtures thereof.
5 . The process according to claim 1 , wherein the reacting to form the compound of Formula (I-1′) is conducted at a temperature in a range of from about −40° C. to about 40° C.
6 . The process according to claim 1 , wherein the at least one Base B is selected from the group consisting of NaOH, KOH, NH 4 OH, and MeNH 2 , and mixtures thereof.
7 . The process according to claim 1 , wherein the at least one Solvent B is selected from the group consisting of MeOH, IPA, and EtOH, and mixtures thereof.
8 . The process according to claim 1 , further comprising isolating the compound of Formula (I) by crystallization from at least one Solvent C selected from the group of IPA, THE, EtOH, and MeCN, and mixtures thereof.
9 . The process according to claim 1 , wherein the at least one Reducing Agent is at least one chemical reducing agent selected from the group consisting of sodium triacetoxy borohydride, sodium cyanoborohydride, sodium borohydride, sodium tris(trifluoroacetoxy) borohydride, tetramethylammonium triacetoxyborohydride, sodium tri (2-methylacetoxy) borohydride, sodium tri (2-phenylacetoxy) borohydride, and mixtures thereof.
10 . The process according to claim 1 , wherein the at least one Reducing Agent is selected from the group consisting of enzymatic reducing agents.
11 . The process according to claim 10 , wherein the at least one Reducing Agent is selected from the group consisting of ketoreductase enzymes.
12 . The process according to claim 10 , wherein the at least one Reducing Agent is selected from the group consisting of ketoreductase enzymes having an amino acid sequence that is at least about 90% identical to the reference sequence of SEQ ID NO: 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11.
13 . The process according to claim 11 , further comprising pre-treating the ketoreductase enzymes with >20% isopropanol.
14 . The process according to claim 1 , further comprising reacting a compound of Formula (I-3) with at least one Fluorinating Agent A in the presence of at least one Catalyst B, at least one Base C, and water to provide the compound of Formula (I-2):
15 . The process according to claim 14 , wherein the at least one Fluorinating Agent A is selected from the group consisting of N-fluorobenzenesulfonimide, 1-chloromethyl-4-fluoro-1,4-diazoniabicyclo[2.2.2] octane bis(tetrafluoroborate), 1-fluoro-4-methyl-1,4-diazoniabicyclo[2.2.2] octanebis(tetrafluoroborate), N-fluoropyridinium triflate, and N-fluoropyridinium tetrafluoroborate, and mixtures thereof.
16 . The process according to claim 14 , wherein the at least one Catalyst B is selected from the group consisting of (C 1 -C 8 alkyl) NH 2 ,
and mixtures thereof.
17 . The process according to claim 14 , wherein the at least one Base C is selected from the group consisting of lithium carbonate, sodium carbonate, sodium bicarbonate, potassium carbonate, ammonium carbonate, cesium carbonate, lithium dihydrogen phosphate, sodium hydrogen phosphate, sodium dihydrogen phosphate, sodium phosphate, potassium hydrogen phosphate, potassium dihydrogen phosphate, potassium phosphate, ammonium hydrogen phosphate, sodium acetate, ammonium acetate, sodium benzoate, sodium hydrogen sulfate, potassium hydrogen sulfate, potassium fluoride, cesium fluoride, sodium formate, and ammonium formate, and mixtures thereof.
18 . The process according to claim 14 , further comprising
reacting a compound of Formula (I-4) with at least one Base D to form a compound of Formula (I-4 1 )
wherein
protecting group PG 1 is selected from the group consisting of isobutyryl, pivaloyl, trityl, tert-butyldiphenylsilyl, and tert-butyldimethylsilyl,
protecting group PG 2 is selected from the group consisting of isobutyryl, pivaloyl, methanesulfonyl, ethanesulfonyl, benzenesulfonyl, 2-nitrobenzenesulfonyl, 4-nitrobenzensulfonyl, 4-chlorobenzenesulfonyl, 4-fluorobenzenesulfonyl, and p-toluenesulfonyl, and
protecting group PG 3 is selected from the group consisting of methanesulfonyl, ethanesulfonyl, benzenesulfonyl, 2-nitrobenzenesulfonyl, 4-nitrobenzene sulfonyl, 4-chlorobenzenesulfonyl, 4-fluorobenzenesulfonyl, and p-toluenesulfonyl, and
the at least one Base D is selected from the group consisting of lithium hydroxide, lithium methoxide, lithium ethoxide, lithium isopropoxide, lithium tert-butoxide, sodium hydroxide, sodium methoxide, sodium ethoxide, sodium isopropoxide, sodium tert-butoxide, and mixtures thereof; and
reacting the compound of Formula (I-4 1 ) with at least one Acid A to form the compound of Formula (I-3)
wherein the at least one Acid A is selected from the group consisting of TFA, HCl, H 2 SO 4 , methanesulfonyl hydroxide, para-toluenesulfonyl hydroxide, and mixtures thereof.
19 . The process according to claim 18 , wherein the reacting the compound of Formula (I-4 1 ) with at least one Acid A to form the compound of Formula (I-3) further comprises reacting the compound of Formula (I-4 1 ) with at least one Tri-Alkyl Amine A, selected from the group consisting of N(C 1 -C 10 alkyl) 3 and mixtures thereof.
20 . The process according to claim 18 , further comprising selectively functionalizing guanosine to provide a compound of Formula (I-4):
wherein protecting group PG 1 is selected from the group consisting of isobutyryl, pivaloyl, trityl, tert-butyldiphenylsilyl, and tert-butyldimethylsilyl; protecting group PG 2 is selected from the group consisting of isobutyryl, pivaloyl, methanesulfonyl, ethanesulfonyl, benzenesulfonyl, 2-nitrobenzenesulfonyl, 4-nitrobenzensulfonyl, 4-chlorobenzenesulfonyl, 4-fluorobenzenesulfonyl, and p-toluenesulfonyl; and protecting group PG 3 is selected from the group consisting of methanesulfonyl, ethanesulfonyl, benzenesulfonyl, 2-nitrobenzenesulfonyl, 4-nitrobenzene sulfonyl, 4-chlorobenzenesulfonyl, 4-fluorobenzenesulfonyl, and p-toluenesulfonyl; wherein the selectively functionalizing guanosine to provide a compound of Formula (I-4) comprises:
a) reacting guanosine with PG 1 -Cl in the presence of at least one Base G, wherein the at least one Base G is selected from the group consisting of pyridine, NMI, 2,6-lutidine, 2,4,6-collidine, DBU, DABCO, tetramethylguanidine, triethylamine, diisopropylethylamine, and mixtures thereof;
b) reacting the product of step a) with PG 2 -Cl and PG 3 -Cl in the presence of at least one Base H, wherein the at least one Base His selected from the group consisting of pyridine, NMI, 2,6-lutidine, 2,4,6-collidine, DBU, DABCO, tetramethylguanidine, triethylamine, diisopropylethylamine, and mixtures thereof; and
c) reacting the product of step b) with R 1 —COCl in the presence of at least one Base I, wherein the at least one Base I is selected form the group consisting of pyridine, NMI, 2,6-lutidine, 2,4,6-collidine, DBU, DABCO, tetramethylguanidine, triethylamine, diisopropylethylamine, and mixtures thereof.
21 . The process according to claim 1 , further comprising
reacting the compound of Formula (I-4) with at least one Base E to form a compound of Formula (I-4 1 )
wherein
protecting group PG 1 is selected from the group consisting of isobutyryl, pivaloyl, trityl, tert-butyldiphenylsilyl, and tert-butyldimethylsilyl chloride,
protecting group PG 2 is selected from the group consisting of isobutyroyl, pivaloyl, methanesulfonyl, ethanesulfonyl, benzenesulfonyl, 2-nitrobenzenesulfonyl, 4-nitrobenzensulfonyl, 4-chlorobenzenesulfonyl, 4-fluorobenzenesulfonyl, and p-toluenesulfonyl, and
protecting group PG 3 is selected from the group consisting of methanesulfonyl, ethanesulfonyl, benzenesulfonyl, 2-nitrobenzenesulfonyl, 4-nitrobenzene sulfonyl, 4-chlorobenzenesulfonyl, 4-fluorobenzenesulfonyl, and p-toluenesulfonyl,
at least one Base E is selected from the group consisting of lithium hydroxide, lithium methoxide, lithium ethoxide, lithium isopropoxide, lithium tert-butoxide, sodium hydroxide, sodium methoxide, sodium ethoxide, sodium isopropoxide, sodium tert-butoxide, and mixtures thereof;
reacting the compound of Formula (I-4 1 ) with at least one Fluorinating Agent B in the presence of at least one Catalyst C, at least one Base F, and water to form a product, followed by reacting the product with at least one Acid B to form a compound of Formula (I-2):
wherein
the at least one Fluorinating Agent B is selected from the group consisting of N-fluorobenzenesulfonimide, 1-chloromethyl-4-fluoro-1,4-diazoniabicyclo[2.2.2] octane bis(tetrafluoroborate), 1-fluoro-4-methyl-1,4-diazoniabicyclo[2.2.2] octanebis(tetrafluoroborate), N-fluoropyridinium triflate, and N-fluoropyridinium tetrafluoroborate, and mixtures thereof,
the at least one Catalyst C is selected from the group consisting of (C 1 -C 8 alkyl) NH 2 ,
and mixtures thereof,
the at least one Base F is selected from the group consisting of lithium carbonate, sodium carbonate, sodium bicarbonate, potassium carbonate, ammonium carbonate, cesium carbonate, lithium dihydrogen phosphate, sodium hydrogen phosphate, sodium dihydrogen phosphate, sodium phosphate, potassium hydrogen phosphate, potassium dihydrogen phosphate, potassium phosphate, ammonium hydrogen phosphate, sodium acetate, ammonium acetate, sodium benzoate, sodium hydrogen sulfate, potassium hydrogen sulfate, potassium fluoride, cesium fluoride, sodium formate, and ammonium formate, and mixtures thereof; wherein the at least one Acid B is selected from the group consisting of TFA, HCl, H 2 SO 4 , methanesulfonyl hydroxide, para-toluenesulfonyl hydroxide, and mixtures thereof; and
the at least one Acid B is selected from the group consisting of TFA, HCl, H 2 SO 4 , MsOH, TsOH, and mixtures thereof.
22 . A compound selected from the group consisting of: