Industrially scalable nucleoside synthesis
View Patent ↗An industrially scalable two-step process for preparing a β-L-2′-deoxy-nucleoside that results in a predominance of the β- over the α-anomeric form of the compound is described. An optional third step may be used to prepare 3′-prodrugs of desirable β-L-2′-deoxy-nucleosides for the delivery of these pharmaceuticals effective for treating viral diseases. The synthetic process is applicable in particular to the formation of β-L-2′-deoxy-cytidine, a pharmaceutically acceptable salt or prodrug thereof. The process can provide a relatively uncontaminated product that may require no further isolation or purification, thereby making the synthesis easily scalable for industrial manufacture.
1. An industrially scalable process for preparing a mixture of α-L-2′-deoxycytidine and β-L-2′-deoxycytidine or a mixture of α-L-thymidine and β-L-thymidine or a pharmaceutically acceptable salt thereof, comprising the steps of:
(a) reacting an N- and/or O-protected cytosine or thymine with a silylating reagent at a temperature from about 10° C. to 50° C., or at ambient temperature, optionally in the presence of a catalyst, to form a silylated cytosine or thymine;
(b) raising the temperature to reflux in order to run the reaction to completion;
(c) cooling the mixture of (a) to about 0° C. to −20° C.;
(d) reacting the silylated cytosine or thymine with a protected 1-halo-2-deoxy-L-sugar in the presence of a Lewis acid at a temperature of about 20° C. to form an N- and/or O-protected deoxycytidine or thymidine mixture;
(e) reacting the N- and/or O-protected deoxycytidine or thymidine anomeric mixture with a deprotecting reagent, to form a mixture of α-L-2′-deoxycytidine and β-L-2′-deoxycytidine or a mixture of α-L-thymidine and β-L-thymidine,
wherein the Lewis acid is selected from SnCl 4 , TiCl 4 , AlCl 3 , ZnCl 2 , CuII, SbCl 5 , and BF 3 (OEt 2 ), wherein said sugar moiety is a protected 2-deoxy-1-L-ribofuranosyl moiety, and wherein the predominate product in each product mixture is the β anomer.
2. The process of claim 1 wherein the final nucleoside product has an α:β anomeric ratio of about 1:5 to 1:6.
3. The process of claim 1 wherein the 1-halo-2-deoxy-L-sugar is a 1-chloro-2-deoxy-L-ribose.
4. The process of claim 1 wherein the 1-halo-2-deoxy-L-sugar is an α-1-halo-2-deoxy-L-ribose.
5. The process of claim 4 wherein the α-1-halo-2-deoxy-L-sugar is an α-1-chloro-2-deoxy-L-ribose.
6. The process of claim 1 wherein the Lewis acid is a Friedel-Crafts catalyst.
7. The process of claim 1 wherein the deprotecting reagent is an alkoxide or ammonia in an alcoholic solvent.
8. An industrially scalable process for preparing a mixture of 2′-deoxy-α-L-cytidine and 2′-deoxy-β-L-cytidine or a pharmaceutically-acceptable salt thereof, comprising the steps of:
(a) reacting an N 4 -protected cytosine with a silylating reagent, optionally in the presence of a catalyst, to form a silylated cytosine;
(b) reacting the silylated cytosine with a protected 1-halo-2-deoxy-L-ribofuranose in the presence of a Lewis acid to form a mixture of N 4 -protected 2′-deoxy-α-L-cytidine and N 4 -protected 2′-deoxy-β-L-cytidine; and
(c) reacting the N 4 -protected 2′-deoxy-α-L-cytidine and N 4 -protected 2′-deoxy-β-L-cytidine mixture with a deprotecting reagent, to form a mixture of 2′-deoxy-α-L-cytidine and 2′-deoxy-β-L-cytidine, wherein the coupling reaction of step (b) is performed at a temperature of about −5° C. or less,
wherein the Lewis acid is selected from SnCl 4 , TiCl 4 , AlCl 3 , ZnCl 2 , CuI, SbCl 5 , and BF 3 (OEt 2 ) and wherein the predominate product in the product mixture is the β anomer.
9. The process of claim 8 wherein the final nucleoside product has an α:β anomeric ratio of about 1:5 to 1:6.
10. The process of claim 8 wherein the 1-halo-2-deoxy-L-ribofuranose is a 1-chloro-2-deoxy-L-ribofuranose.
11. The process of claim 8 wherein the 1-halo-2-deoxy-L-ribofuranose is an α1-halo-2-deoxy-L-ribofuranose.
12. The process of claim 11 wherein the α-1-halo-2-deoxy-L-ribofuranose is an α-1-chloro-2-deoxy-L-ribofuranose.
13. The process of claim 8 wherein the Lewis acid is a Friedel-Crafts catalyst.
14. The process of claim 13 wherein the Friedel-Crafts catalyst is TiCl 4 .
15. The process of claim 8 further comprising directly esterifying 2′-deoxy-α-L-cytidine and/or 2′-deoxy-β-L-cytidine with an amino acid at the 3′-position in the presence of a polar aprotic solvent and a coupling agent selected from the group consisting of EDC (1-[3-(dimethylamino)-propyl]-3-ethyl-carbodiimide hydroxide), carbonyldiimidazole (CDI), BOP reagent (benzotriazol-1-yloxy-tris(dimethylamino)-phosphonium hexafluorophosphate, and diethyl azodicarboxylate in the presence of triphenylphosphine (Mitsunobu reaction), to prepare a 3′-ester of 2′-deoxy-α-L-cytidine and/or 2′-deoxy-β-L-cytidine.
16. An industrially scalable process for preparing a mixture of 2′-deoxy-α-L-cytidine and 2′-deoxy-β-L-cytidine or a pharmaceutically-acceptable salt thereof, comprising the steps of:
(a) reacting an N 4 -benzoylcytosine with a silylating agent, to form silylated N 4 -benzoylcytosine;
(b) adding 1-α-chloro-3′,5′-di-O-toluoyl-2′-deoxy-L-ribofuranose, in the presence of a Lewis acid, to the silylated N 4 -benzoylcytosine to form a mixture of 3′,5′-di-O-toluoyl-2′-deoxy-α-L- and 3′,5′-di-O-toluoyl-2′-deoxy-β-L-cytidines; and
(c) deprotecting the mixture of 3′,5′-di-O-toluoyl-2′-deoxy-αL-cytidine and 3′,5′-di-O-toluoyl-2′-deoxy-β-L-cytidine, wherein the coupling reaction of step (b) is performed at a temperature of about −5° C or less,
wherein the Lewis acid is selected from SnCl 4 , TiCl 4 , AlCl 3 , ZnCl 2 , CuI, SbCl 5 , and BF 3 (OEt 2 ) and wherein the predominate product in the product mixture is the β anomer.
17. The process of claim 16 wherein the final nucleoside product has an α:β anomeric ratio of about 1:5 to 1:6.
18. The process of claim 16 wherein the silylating agent is N,O-bis-(trimethylsilyl)-acetamide.
19. The process of claim 16 wherein the Lewis acid is a Friedel-Crafts catalyst.
20. The process of claim 19 wherein the Friedel-Crafts catalyst is TiCl 4 .
21. The process of claim 16 wherein the reaction is run in dichloromethane.
22. The process of claim 16 wherein the 3′,5′-di-O-toluoyl-2′-deoxy-β-L-cytidine is deprotected by using sodium methoxide in methanol.
23. The process of claim 16 further comprising directly esterifying 2′-deoxy-α-L-cytidine and/or 2′-deoxy-β-L-cytidine with an amino acid at the 3′-position in the presence of a polar aprotic solvent and a coupling agent selected from the group consisting of EDC (1-[3-(dimethylamino)-propyl]-3-ethyl-carbodiimide hydroxide), carbonyldiimidazole (CDl), BOP reagent (benzotriazol-1 -yloxy-tris(dimethylamino)-phosphonium hexafluorophosphate, and diethyl azodicarboxylate in the presence of triphenylphosphine (Mitsunobu reaction), to prepare a 3′-ester of 2′-deoxy-α-L-cytidine and/or 2′-deoxy-β-L-cytidine.