Polymer derivatives

• US 20050085619A1
• Filed: 10/07/2004
• Published: 04/21/2005
• 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.
• Granted Patent: US 7,230,068 B2
• Time in Patent Office: Days
• US Class Current: 528/405
• 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 and at least one terminal azide moiety, wherein said polymer is selected from the group consisting of poly(alkylene oxides), poly(oxyethylated polyols), and poly(olefinic alcohols).

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), poly(oxyethylated glucose), and poly(vinyl alcohol).

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 3, wherein said compound comprises poly(ethylene glycol)azide and the azide moiety is covalently attached to a polymer backbone via a linker.

6. 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.

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

8. 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 can be the same or different from said azide moiety.

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

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

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

12. The water-soluble compound of claim 1, wherein said azide moiety forms a linkage with a reactive moiety on a surface or in a molecule.

13. The water-soluble compound of claim 12, wherein said reactive moiety is an acetylene moiety.

14. 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.

15. The water-soluble compound of claim 14, 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.

16. The water-soluble compound of claim 14, wherein said second functional group is amine-specific and said linker moiety includes an active amine moiety.

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

18. The water-soluble compound of claim 14, wherein said second functional group is specific for reaction with an activated ester and said linker moiety includes an activated ester.

19. The water-soluble compound of claim 14, wherein said second functional group is specific for reaction with a ketone and said linker moiety includes an ketone.

20. The water-soluble compound of claim 14, wherein said second functional group is specific for reaction with a thiol nucleophile and said linker moiety includes a thiol nucleophile.

21. The water-soluble compound of claim 14 wherein said second functional group is specific for reaction with a hydroxyl nucleophile and said linker moiety includes a hydroxyl nucleophile.

22. The water-soluble compound of claim 14 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—

.

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

24. 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 can be the same or different from the azide.

25. The water soluble activated polymer of claim 24 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, and tresylate, alkene, and ketone.

26. The water soluble activated polymer of claim 24 wherein n is from about 5 to 2,200.

27. The water soluble activated polymer of claim 24 wherein n is from about 34 to 1,100.

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

29. The water soluble activated polymer of claim 24 wherein R is methoxy (CH₃O—

).

30. The water soluble activated polymer of claim 24 wherein R is carboxylic acid (HO₂C—

).

31. The water soluble activated polymer of claim 24 wherein R is amine (H₂N—

).

32. The water soluble activated polymer of claim 24 wherein R is maleimide.

33. The water soluble activated polymer of claim 24 wherein R is reactive with amines.

34. The water soluble activated polymer of claim 24 wherein R is reactive with an electrophilic carbonyl group.

35. The water soluble activated polymer of claim 24 wherein R is reactive with thiols.

36. The water soluble activated polymer of claim 24 wherein R is reactive with hydroxyls.

37. A water-soluble compound comprising a polymer and at least one acetylene moiety, wherein said polymer is selected from the group consisting of poly(alkylene oxides), poly(oxyethylated polyols), and poly(olefinic alcohols).

38. The water-soluble compound of claim 37 wherein said acetylene moiety is a terminal acetylene group.

39. 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).

40. The water-soluble compound of claim 37 wherein said polymer is poly(ethylene glycol).

41. The water-soluble compound of claim 40 wherein said compound is poly(ethylene glycol)acetylene and said acetylene moiety is covalently attached directly to a polymer backbone.

42. The water-soluble compound of claim 40, wherein said polymer is poly(ethylene glycol)acetylene and wherein the acetylene moiety is covalently linked to a polymer backbone via a linker moiety.

43. The water-soluble compound of claim 40 wherein said polymer is poly(ethylene glycol)acetylene, wherein said acetylene moiety is covalently bound to a polymer backbone via an ether linkage.

44. The water-soluble compound of claim 37 wherein said polymer is a straight chain polymer that is not substituted beyond the acetylene moiety.

45. The water-soluble compound of claim 37 wherein said polymer is a random or block copolymer or terpolymer.

46. The water-soluble compound of claim 37 wherein said compound comprises a dumbbell structure comprising:

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

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

wherein the second functional group can be the same or different from said acetylene moiety.

47. The water-soluble compound of claim 46, wherein said second functional group is not reactive with acetylene moieties.

48. The water-soluble compound of claim 37 wherein said polymer comprises at least one arm of a branched molecular structure.

49. The water-soluble compound of claim 48 wherein said branched molecular structure is dendritic.

50. The water-soluble compound of claim 46 wherein said acetylene moiety forms a linkage with a reactive moiety on a surface or in a molecule.

51. The water-soluble compound of claim 46 wherein said reactive moiety is an acetylene moiety.

52. The water-soluble compound of claim 37 wherein said acetylene 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 acetylene moiety for linking to said linker moiety.

53. The water-soluble compound of claim 46 wherein said second functional group is specific for nucleophilic displacement and said linker moiety comprises a nucleophilic moiety capable of reacting with such active moiety.

54. The water-soluble compound of claim 52 wherein said second functional group is amine-specific and said linker moiety includes an active amino moiety.

55. The water-soluble compound of claim 52 wherein said second functional group is specific for reaction at an electrophilic carbonyl and said linker moiety includes an electrophilic carbonyl moiety.

56. The water-soluble compound of claim 52 wherein said second functional group is specific for reaction with an activated ester and said linker moiety includes an activated ester.

57. The water-soluble compound of claim 52 wherein said second functional group is specific for reaction with an ketone and said linker moiety includes an ketone.

58. The water-soluble compound of claim 52 wherein said second functional group is specific for reaction with a thiol nucleophile and said linker moiety includes a thiol nucleophile.

59. The water-soluble compound of claim 52 wherein said second functional group is specific for reaction with a hydroxyl nucleophile and said linker moiety includes a hydroxyl nucleophile.

60. The water-soluble compound of claim 52 wherein said linker moiety is selected from the group consisting of:

—

NH—

CO—

CH₂—

CH₂—

; and

—

CO—

NH—

CH₂—

CH₂—

.

61. The water-soluble compound of claim 37 wherein said activated polymer is stable in aqueous environments at a pH of about 11 or less.

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

R—

CH₂CH₂—

(OCH₂CH₂)_n—

C≡

CH

wherein n is from about 5 to 3,000, and R is a capping group, a functional group, or a leaving group that can be the same or different from the azide.

63. The water soluble activated polymer of claim 62 wherein X is 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, and tresylate, alkene, and ketone.

64. The water soluble activated polymer of claim 62 wherein n is from about 5 to 2,200.

65. The water soluble activated polymer of claim 62 wherein n is from about 34 to 1,100.

66. The water soluble activated polymer of claim 62 wherein n is from about 45 to 110.

67. The water soluble activated polymer of claim 62 wherein R is methoxy (CH₃O—

).

68. The water soluble activated polymer of claim 62 wherein R is carboxylic acid (HO₂C—

).

69. The water soluble activated polymer of claim 62 wherein R is amine (H₂N—

).

70. The water soluble activated polymer of claim 62 wherein R is maleimide.

71. The water soluble activated polymer of claim 62 wherein R is reactive with amines.

72. The water soluble activated polymer of claim 62 wherein R is reactive with electrophilic carbonyl groups.

73. The water soluble activated polymer of claim 62 wherein R is reactive with thiols.

74. The water soluble activated polymer of claim 62 wherein R is reactive with hydroxyls.