Non-enzymatic synthesis of O-acetyl-ADP-ribose and analogues thereof

Patent metadata

Field	Value
Publication number	US-9376461-B2
Application number	US-201214124395-A
Country	US
Kind code	B2
Filing date	Jun 6, 2012
Priority date	Jun 6, 2011
Publication date	Jun 28, 2016
Grant date	Jun 28, 2016

Abstract

Provided herein is a simple, one-step, non-enzymatic synthesis of O-Acetyl-ADP-ribose (OAADPR) from NAD and sodium acetate in acetic acid. The extension of this reaction to other carboxylic acids, demonstrates that the reaction between NAD, and NAD analogs produces mixtures of the corresponding 2′- and 3′-carboxylic esters. Included are O-carboxyl-ADP-ribose compounds and corresponding methods of synthesis (e.g., O-propionyl-ADP-ribose, O-succinyl-ADP-ribose, O-malonyl-ADP-ribose), as well as non-adenosine nucleoside compounds.

First claim

We claim:

1. A method of non-enzymatic synthesis of compound (III), a 2′-O— or 3′-O-carboxylic ester of the ribose moiety of adenosine-5′-diphosphate ribose (ADP-ribose) or a 2′-O— or 3 ‘-O-carboxylic ester of the ribose moiety of a non-adenosine nucleoside-5’-diphosphate ribose (NDP-ribose) analog, comprising reacting compound (I) defined as
(i) nicotinamide adenine dinucleotide (NAD) or a non-adenosine nucleoside-5′-diphosphate ribose (NDP-ribose) analog (I) wherein the non-adenosine substituent moiety is selected from the group consisting of 5′-guanosinyl, 5′-(5-methyluridinyl), 5′-uridinyl, and 5′-cytidinyl; and
(ii) the variable LG (leaving group) is selected from the group consisting of a 1-nicotinamidinyl moiety, a 1-nicotinic acid-derived moiety and a halide ion;
with a carboxylate ion (II) to produce a mixture of the two O-acylated compounds (III a) and (III b)

wherein one of R 1 and R 2 is H, and the other is —C(O)—R x such that the product (III) comprises a mixture of 2′-O-carboxyl-ADP/NDP-ribose (III a, wherein R 1 is H and R 2 is —C(O)—R x ) and 3′-O-carboxyl-ADP/NDP-ribose (III b, wherein R 1 is —C(O)—R x and R 2 is H), wherein R x is an optionally substituted C 1 -C 12 straight or branched alkyl or
alkenyl moiety, or an optionally substituted carbocyclyl moiety that is saturated, unsaturated, or aromatic 5-7 membered monocyclic ring or 8-12 membered bicyclic ring, or an
optionally substituted heterocyclyl moiety that is a saturated, unsaturated, or aromatic 5-7 membered monocyclic ring or 8-12 membered bicyclic rings comprising one or more heteroatoms selected from N,O, and S.

2. The method of claim 1 , wherein (I) is NAD.

3. The method of claim 2 , wherein the carboxylate (II) is acetate and the product is a mixture of 2′-O-acetyl ADP-ribose (III a) and 3′-O-acetyl ADP-ribose (III b).

4. The method of claim 1 , wherein the carboxylate (II) is selected from the group consisting of propionate, n-butyrate, isobutyrate, trimethylacetate, trans-2-butenoate, n-pentanoate, 3,3-dimethylacrylate, n-hexanoate, n-heptanoate, benzoate, succinate, citrate, DL-lactate and L-malate.

5. The method of claim 1 , wherein the leaving group is a halide ion selected from the group consisting of Cl − , Br − , I − and F − .

6. The method of claim 1 , wherein the carboxylate is reacted with the nicotinamide adenine dinucleotide (NAD) or analog thereof at a temperature of 20° C. to 120° C. in a reaction mixture optionally further comprising an organic solvent.

7. The method of claim 6 , further comprising wherein the carboxylate is formed by the reaction of an inorganic or an organic base with a carboxylic acid before the addition of the NAD or analog thereof.

8. The method of claim 7 , wherein the inorganic base is Na 2 CO 3 or NaHCO 3 .

9. The method of claim 7 , wherein the organic base is selected from the group consisting of a trialkylamine, a 5 to 6-membered nitrogen containing heterocycle, and an optionally C 1 -C 4 substituted 5 to 6-membered nitrogen containing heterocycle.

10. The method of claim 7 , wherein the carboxylic acid is selected from the group consisting of propionic acid, n-butyric acid, isobutyric acid, trimethylacetic acid, trans-butenoic acid, n-pentanoic acid, 3, 3-dimethylacrylic acid, n-hexanoic acid, n-heptanoic acid, benzoic acid, succinic acid, citric acid, DL-lactic acid, and L-malic acid.

11. The method of claim 7 , wherein the carboxylic acid is acetic acid.

12. The method of claim 6 , wherein the organic solvent is selected from the group consisting of dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidinone, trimethyl phosphate, triethyl phosphate, formamide and dimethylsulfone.

13. The method of claim 6 , wherein the temperature is about 85° C. to 95° C.

14. The method of claim 7 , wherein the inorganic base is Na 2 CO 3 , the nicotinamide adenine dinucleotide (NAD) or analog thereof is NAD, and the carboxylic acid is acetic acid.

15. The method of claim 7 , wherein the organic base is pyridine, the nicotinamide adenine dinucleotide (NAD) or analog thereof is NAD and the carboxylic acid is selected from the group consisting of succinic acid, citric acid, DL-lactic acid and L-malic acid.

16. The method of claim 1 , further comprising the step of purifying the product by ion chromatography.

17. The method of claim 6 , further comprising the step of purifying the product by ion chromatography.

18. The method of claim 16 , further comprising wherein ion chromatography is performed on an aminopropyl functionalized silica gel column.

19. The method of claim 1 , further comprising wherein when R x is optionally substituted, it is optionally substituted with one or more substituents selected from the group consisting of a halogen, an alkyl, an amine, a hydroxyl, an O-alkyl, a N-alkyl, an aryl, an arylalkyl, a cycloalkyl, a cycloheteroalkyl, a heteroalkyl, a heteroaryl, and a heteroarylalkyl.

20. A method for the non-enzymatic synthesis of a mixture of (III a) and (III b) that are 2′-O— and 3′-O-carboxylic acid esters of the ribose moiety of adenosine diphosphate ribose (ADP-ribose) comprising
reacting compound (IV) defined as
nicotinamide adenine dinucleotide (NAD) wherein Y is a 1-nicotinamidyl moiety, or an analog thereof wherein Y is a 1-nicotinic acid acid-derived moiety;
with a carboxylate ion (II) to produce the O-acylated ADP-ribose products defined by formulas (III a) and (III b),

wherein the product (III, wherein one of R 1 and R 2 is H, and the other is —C(O)—R x ) comprises a mixture of 2′-O-acyl-ADP-ribose (III a, wherein R 1 is H and R 2 is —C(O)—R x ) and 3′-O-acyl-ADP-ribose (III b, wherein R 1 is —C(O)—R x and R 2 is H), wherein R x is an optionally substituted C 1 -C 12 straight or branched alkyl or alkenyl moiety, or an optionally substituted carbocyclyl moiety that is a saturated, unsaturated, or aromatic 5-7 membered monocyclic ring or 8-12 membered bicyclic ring, or an optionally substituted heterocyclyl moiety that is a saturated, unsaturated, or aromatic 5-7 membered monocyclic ring or 8-12 membered bicyclic rings comprising one or more heteroatoms selected from N, O, and S.

21. The method of claim 20 , wherein Y is 1-nicotinamidyl.

22. The method of claim 20 , wherein Y is a 1-nicotinic acid-derived moiety.

23. The method of claim 20 , wherein the carboxylate is acetate and the product is O-acetyl-ADP ribose comprising a mixture of 2′-O-acetyl-ADP-ribose (III a) and 3′-O-acetyl-ADP-ribose (III b).

24. The method of claim 20 , wherein the carboxylate is formed by the reaction of an inorganic base with a carboxylic acid before the addition of the NAD or analog thereof.

25. The method of claim 24 , wherein the inorganic base is Na 2 CO 3 or NaHCO 3 .

26. The method of claim 20 , wherein the carboxylate (III) is formed by the reaction with acetic acid.

27. The method of claim 20 , wherein the nicotinamide adenine dinucleotide (NAD) or analog thereof is NAD.

28. The method of claim 20 , wherein the reaction temperature is about 85° C. to 95° C.

29. The method of claim 20 , wherein the carboxylate is formed by the reaction of a carboxylic acid and Na

Assignees

• Koppetsch Karsten
• Szczepankiewicz Bruce G
• Perni Robert B
• Glaxosmithkline Llc

Inventors

• Koppetsch Karsten
• Szczepankiewicz Bruce G
• Perni Robert B

Classifications

• C07H13/04

  having the esterifying carboxyl radicals attached to acyclic carbon atoms · CPC title

• C07H1/00 Primary

  Processes for the preparation of sugar derivatives · CPC title

• C07H19/20 Primary

  with the saccharide radical esterified by phosphoric or polyphosphoric acids · CPC title