Polymer derivatives

• US 7,230,068 B2
• Filed: 10/07/2004
• Issued: 06/12/2007
• Est. Priority Date: 10/09/2003
• Status: Active Grant

• Current Assignee: Ambrx, Inc. (Johnson & Johnson)
• Original Assignee: Ambrx, Inc. (Johnson & Johnson)
• Inventors: Wilson, Troy E.
• Primary Examiner(s): SELLERS, ROBERT E
• Application Number:
  US10/960,674
• Publication Number:
  US 20050085619A1
• Time in Patent Office: 978 Days
• Field of Search: 514/772.4, 514/772.7, 525/54, 525/11, 525/61, 525/404, 525/408, 525/409, 528/421
• US Class Current: 528/407
• CPC Class Codes: C08F 216/16 Monomers containing no hetero atoms other than the ether oxygen
• CPC Class Codes: C08F 8/02 Alkylation
• CPC Class Codes: C08F 8/30 Introducing nitrogen atoms or nitrogen-containing groups
• CPC Class Codes: C08G 65/325 containing nitrogen
• CPC Class Codes: C08G 65/329 with organic compounds
• CPC Class Codes: C08G 65/333 containing nitrogen

Abstract

The invention provides water-soluble compounds that include a polymer and at least one terminal azide or acetylene moiety. Also provided are highly efficient methods for the selective modification of proteins with PEG derivatives, which involves the selective incorporation of non-genetically encoded amino acids, e.g., those amino acids containing an azide or acetylene moiety, into proteins in response to a selector codon and the subsequent modification of those amino acids with a suitably reactive PEG derivative.

Claims

1. A water-soluble compound comprising a polymer containing at least one terminal azide moiety and at least one terminal non-azide moiety selected from the group consisting of hydroxyl, protected hydroxyl, alkoxyl, active ester, active carbonate, acetal, aldehyde, aldehyde hydrates, alkenyl, acrylate, methacrylate, acrylamide, active sulfone, amine, aminooxy, protected amine, hydrazide, protected hydrazide, protected thiol, protected carboxylic acid, isocyanate, isothiocyanate, maleimide, vinylsulfone, dithiopyridine, vinylpyridine, iodoacetamide, epoxide, glyoxals, diones, mesylates, tosylates, tresylate, alkene, and ketone, wherein said polymer is selected from the group consisting of poly(alkylene oxides) and poly(oxyethylated polyols), wherein said compound has a molecular weight selected from the group consisting of between about 6000 and about 80,000, between about 6,000 and 20,000, between about 80,000 and about 100,000, about 6,000, about 20,000, about 80,000, and about 100,000.

2. The water-soluble compound of claim 1, wherein said polymer is selected from the group consisting of poly(ethylene glycol), poly(propylene glycol), poly(oxyethylated glycerol), poly(oxyethylated sorbitol), and poly(oxyethylated glucose).

3. The water-soluble compound of claim 1, wherein said polymer comprises poly(ethylene glycol).

4. The water-soluble compound of claim 3, wherein said compound comprises poly(ethylene glycol)azide and the azide moiety is covalently attached directly to a polymer backbone.

5. The water-soluble compound of claim 1, wherein said polymer comprises a straight chain polymer and the compound is not substituted beyond the azide moiety.

6. The water-soluble compound of claim 1, wherein said polymer is a random or block copolymer or terpolymer.

7. The water-soluble compound of claim 1, wherein said compound comprises a dumbbell structure comprising:

a) said azide moiety on at least a first end of a polymer backbone; and

b) at least a second functional group on a second end of the polymer backbone;

wherein the second functional group is different from said azide moiety.

8. The water-soluble compound of claim 7, wherein said second functional group is not reactive with azide moieties.

9. The water-soluble compound of claim 1, wherein said compound comprises at least one arm of a branched molecular structure.

10. The water-soluble compound of claim 9, wherein said branched molecular structure is dendritic.

11. The water-soluble compound of claim 1, wherein said azide moiety is linked to said polymer by a linkage that comprises a linker moiety and wherein said polymer comprises at least a second functional group other than said azide moiety for linking to said linker moiety.

12. The water-soluble compound of claim 11, wherein said second functional group is specific for nucleophilic displacement and said linker moiety includes a nucleophilic moiety capable of reacting with said functional group.

13. The water-soluble compound of claim 11, wherein said second functional group is amine-specific and said linker moiety includes an active amine moiety.

14. The water-soluble compound of claim 11, wherein said second functional group is specific for reaction at an electrophilic carbonyl and said linker moiety includes an electrophilic carbonyl moiety.

15. The water-soluble compound of claim 11, wherein said second functional group is specific for reaction with an activated ester and said linker moiety includes an activated ester.

16. The water-soluble compound of claim 11, wherein said second functional group is specific for reaction with a ketone and said linker moiety includes an ketone.

17. The water-soluble compound of claim 11, wherein said second functional group is specific for reaction with a thiol nucleophile and said linker moiety includes a thiol nucleophile.

18. The water-soluble compound of claim 11 wherein said second functional group is specific for reaction with a hydroxyl nucleophile and said linker moiety includes a hydroxyl nucleophile.

19. The water-soluble compound of claim 11 wherein said linker moiety is selected from the group consisting of:

—

NH—

CO—

CH₂—

CH₂—

;

—

CO—

NH—

CH₂—

CH₂—

;

—

S—

CH₂—

CH₂—

; and

—

O—

CH₂—

CH₂—

.

20. The water-soluble compound of claim 1, wherein said compound is stable in aqueous environments at a pH of about 11 or less.

48. The water-soluble compound of claim 1, wherein said at least one terminal non-azide moiety is a N-hydroxysuccinimidyl ester.

49. The water-soluble compound of claim 1, wherein said at least one terminal non-azide moiety is a 1-benzotriazolyl ester.

50. The water-soluble compound of claim 1, wherein said at least one terminal non-azide moiety is a N-hydroxysuccinimidyl carbonate.

51. The water-soluble compound of claim 1, wherein said at least one terminal non-azide moiety is a 1-benzotriazolyl carbonate.

21. A water soluble activated polymer that is stable against hydrolysis, said activated polymer having the formula:

R—

CH₂CH₂—

(OCH₂ CH₂)_n—

N₃;

wherein

n is from about 5 to 3,000, and R is a capping group, a functional group, or a leaving group that is different from the azide, wherein R is a functional group selected from the group consisting of hydroxyl, protected hydroxyl, alkoxyl, N-hydroxysuccinimidyl ester, 1-benzotriazolyl ester, N-hydroxysuccinimidyl carbonate, 1-benzotriazolyl carbonate, acetal, aldehyde, aldehyde hydrates, alkenyl, acrylate, methacrylate, acrylamide, active sulfone, amine, aminooxy, protected amine, hydrazide, protected hydrazide, protected thiol, carboxylic acid, protected carboxylic acid, isocyanate, isothiocyanate, maleimide, vinylsulfone, dithiopyridine, vinylpyridine, iodoacetamide, epoxide, glyoxals, diones, mesylates, tosylates, tresylate, alkene, and ketone.

22. The water soluble activated polymer of claim 21 wherein n is from about 5 to 2,200.

23. The water soluble activated polymer of claim 21 wherein n is from about 34 to 1,100.

24. The water soluble activated polymer of claim 21 wherein n is from about 45 to 110.

25. The water soluble activated polymer of claim 21 wherein R is carboxylic acid (HO₂C—

).

26. The water soluble activated polymer of claim 21 wherein R is amine (H₂N—

).

27. The water soluble activated polymer of claim 21 wherein R is maleimide.

28. The water soluble activated polymer of claim 21 wherein R is reactive with amines.

29. The water soluble activated polymer of claim 21 wherein R is reactive with an electrophilic carbonyl group.

30. The water soluble activated polymer of claim 21 wherein R is reactive with thiols.

31. The water soluble activated polymer of claim 21 wherein R is reactive with hydroxyls.

32. A water soluble compound comprising a polymer containing at least one terminal azide moiety and at least one terminal methoxy moiety, wherein said polymer is selected from the group consisting of poly(alkylene oxides), poly(oxyethylated polyols), and poly(olefinic alcohols), and wherein said polymer has a molecular weight between about 20 kDa and about 100 kDa.

33. The water-soluble compound of claim 32, wherein said polymer is selected from the group consisting of poly(ethylene glycol), poly(propylene glycol), poly(oxyethylated glycerol), poly(oxyethylated sorbitol), poly(oxyethylated glucose), and poly(vinyl alcohol).

34. The water-soluble compound of claim 32, wherein said polymer comprises poly(ethylene glycol).

35. A water soluble compound comprising a polymer and at least one terminal azide moiety and at least one terminal methoxy moiety, wherein said polymer has a molecular weight of about 20 kDa.

36. A water soluble compound comprising a polymer and at least one terminal azide moiety and at least one terminal methoxy moiety, wherein said polymer has a molecular weight of about 100 kDa.

37. A water soluble compound comprising a polymer containing at least one terminal azide moiety and at least one terminal methoxy moiety, wherein said polymer is selected from the group consisting of poly(alkylene oxides), poly(oxyethylated polyols), and poly(olefinic alcohols), and wherein said polymer has a molecular weight between about 6 kDa and about 80 kDa.

38. The water-soluble compound of claim 37, wherein said polymer is selected from the group consisting of poly(ethylene glycol), poly(propylene glycol), poly(oxyethylated glycerol), poly(oxyethylated sorbitol), poly(oxyethylated glucose), and poly(vinyl alcohol).

39. The water-soluble compound of claim 37, wherein said polymer comprises poly(ethylene glycol).

40. A water soluble compound comprising a polymer and at least one terminal azide moiety and at least one terminal methoxy moiety, wherein said polymer has a molecular weight of about 6 kDa.

41. A water soluble compound comprising a polymer containing at least one terminal azide moiety and at least one terminal methoxy moiety, wherein said polymer has a molecular weight of about 80 kDa.

42. A water soluble compound comprising a polymer containing at least one terminal azide moiety and at least one terminal methoxy moiety, wherein said polymer is selected from the group consisting of poly(alkylene oxides), poly(oxyethylated polyols), and poly(olefinic alcohols), and wherein said polymer has a molecular weight between about 6 kDa and about 20 kDa.

43. The water-soluble compound of claim 42, wherein said polymer is selected from the group consisting of poly(ethylene glycol), poly(propylene glycol), poly(oxyethylated glycerol), poly(oxyethylated sorbitol), poly(oxyethylated glucose), and poly(vinyl alcohol).

44. The water-soluble compound of claim 42, wherein said polymer comprises poly(ethylene glycol).

45. A water soluble compound comprising a polymer containing at least one terminal azide moiety and at least one terminal methoxy moiety, wherein said polymer is selected from the group consisting of poly(alkylene oxides), poly(oxyethylated polyols), and poly(olefinic alcohols), and wherein said polymer has a molecular weight between about 80 kDa and about 100 kDa.

46. The water-soluble compound of claim 45, wherein said polymer is selected from the group consisting of poly(ethylene glycol), poly(propylene glycol), poly(oxyethylated glycerol), poly(oxyethylated sorbitol), poly(oxyethylated glucose), and poly(vinyl alcohol).

47. The water-soluble compound of claim 45, wherein said polymer comprises poly(ethylene glycol).