N-maleimidyl polymer derivatives

• US 6,911,197 B2
• Filed: 04/07/2003
• Issued: 06/28/2005
• Est. Priority Date: 02/22/2000
• Status: Expired due to Term

• Current Assignee: Nektar Therapeutics
• Original Assignee: Nektar Therapeutics Al Corporation (Nektar Therapeutics)
• Inventors: Shen, Xiaoming
• Primary Examiner(s): Spear, James M.
• Assistant Examiner(s): FUBARA, BLESSING M
• Application Number:
  US10/408,834
• Publication Number:
  US 20040109841A1
• Time in Patent Office: 813 Days
• Field of Search: 424/78.08, 424/78.31, 424/78.32, 424/78.33, 525/404
• US Class Current: 424/78.08
• CPC Class Codes: A61K 31/74 Synthetic polymeric materials
• CPC Class Codes: A61K 47/58 obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. poly[meth]acrylate, polyacrylamide, polystyrene, polyvinylpyrrolidone, polyvinylalcohol or polystyrene sulfonic acid resin
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61K 47/65 Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers
• CPC Class Codes: C08G 65/00 Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
• CPC Class Codes: C08G 65/33337 cyclic
• CPC Class Codes: C08G 65/33396 having oxygen in addition to nitrogen
• CPC Class Codes: C08G 65/48 Polymers modified by chemical after-treatment

Abstract

The invention is directed to multi-functional N-maleimidyl polymer derivatives comprising a water soluble and non-peptidic polymer backbone having a terminal carbon, such as a poly(alkylene glycol), the terminal carbon of the polymer backbone being directly bonded to the nitrogen atom of a N-maleimidyl moiety without a linking group therebetween. The invention also provides two methods of preparing such linkerless N-maleimidyl polymer derivatives.

Claims

1. A polymer conjugate comprising a multi-functional polymer attached to at least one biologically active agent, said polymer comprising a water soluble and non-peptidic polymer backbone having an average molecular weight from about 800 Da to about 100,000 Da, the polymer backbone having a first terminus bonded to a first functional group and a second terminus having a terminal carbon, wherein said terminal carbon of said second terminus is directly bonded to a N-maleimidyl moiety having the structure:

wherein L is the point of bonding to the terminal carbon of the second terminus of the polymer backbone, said first functional group being different from said N-maleimidyl moiety, and wherein said biologically active agent is attached to the polymer via reaction with the N-maleimidyl moiety or the first functional group.

2. The conjugate of claim 1, wherein said water soluble and non-peptidic polymer backbone has from about 2 to about 300 termini.

3. The conjugate of claim 1, wherein said water soluble and non-peptidic polymer backbone is selected from the group consisting of poly(alkylene glycol), copolymers thereof, terpolymers thereof, and mixtures thereof.

4. The conjugate of claim 1, wherein said water soluble and non-peptidic polymer backbone is selected from the group consisting of poly(ethylene glycol), poly(propylene glycol), and copolymers of ethylene glycol and propylene glycol.

5. The conjugate of claim 1, wherein the polymer has the structure:

wherein;

POLY is a linear water soluble and non-peptidic polymer having a terminal carbon, the terminal carbon being directly bonded to the nitrogen atom of the N-maleimidyl moiety; and

X is said first functional group.

6. The conjugate of claim 5, wherein POLY is selected from the group consisting of poly(alkylene glycol), copolymers thereof, terpolymers thereof, and mixtures thereof.

7. The conjugate of claim 5, wherein POLY is selected from the group consisting of poly(ethylene glycol), poly(propylene glycol), and copolymers of ethylene glycol and propylene glycol.

8. The conjugate of claim 5, wherein POLY is poly(ethylene glycol) having an average molecular weight from about 6,000 Da to about 80,000 Da.

9. The conjugate of claim 5, wherein X is selected from the group consisting of hydroxyl, protected hydroxyl, active ester, active carbonate, acetal, aldehyde, aldehyde hydrate, alkenyl, acrylate, methacrylate, acrylarnide, active sulfone, amine, protected amine, hydrazide, protected hydrazide, protected thiol, carboxylic acid, protected carboxylic acid, isocyanate, isothiocyanate, vinylsulfone, dithiopyridine, vinylpyridine, iodoacetamide, epoxide, glyoxal, dione, mesylate, tosylate, and tresylate.

10. The conjugate of claim 5, wherein X is selected from the group consisting of —

OH, —

NH₂, —

CO₂H, —

CHO, —

CH(OC₂H₅)₂, N-hydroxysuccinimidyl ester, 1-benzotriazolyl ester, N-hydroxysuccinimidyl carbonate, 1-benzotriazolyl carbonate, and tresylate.

11. The conjugate of claim 1, wherein the water soluble and non-peptidic polymer backbone has the structure:

X—

CH₂CH₂O—

(CH₂CH₂O)_n—

CH₂CH₂—

L

wherein;

L is the point of bonding to the nitrogen atom of the N-maleimidyl moiety;

X is said first functional group; and

n is about 20 to about 4,000.

12. The conjugate of claim 11, wherein X is selected from the group consisting of hydroxyl, protected hydroxyl, active ester, active carbonate, acetal, aldehyde, aldehyde hydrate, alkenyl, acrylate, methacrylate, acrylamide, active sulfone, amine, protected amine, hydrazide, protected hydrazide, protected thiol, carboxylic acid, protected carboxylic acid, isocyanate, isothiocyanate, vinylsulfone, dithiopyridine, vinylpyridine, iodoacetamide, epoxide, glyoxal, dione, mesylate, tosylate, and tresylate.

13. The conjugate of claim 11, wherein X is selected from the group consisting of —

OH, —

NH₂, —

CO₂H, —

CHO, —

CH(OC₂H₅)₂, N-hydroxysuccinimidyl ester, 1-benzotriazolyl ester, N-hydroxysuccinimidyl carbonate, 1-benzotriazolyl carbonate, and tresylate.

14. The conjugate of claim 11, wherein n is about 20 to about 2,000.

15. The conjugate of claim 1, wherein the polymer backbone has an average molecular weight from about 6,000 Da to about 80,000 Da.

16. The conjugate of claim 1, wherein the polymer backbone is poly(ethylene glycol) having an average molecular weight from about 6,000 Da to about 80,000 Da.

17. The conjugate of claim 1, wherein the polymer backbone is selected from the group consisting of linear poly(ethylene glycol), branched poly(ethylene glycol), forked poly(ethylene glycol), pendant poly(ethylene glycol), and poly(ethylene glycol) having degradable linkages therein.

18. The conjugate of claim 1, wherein the at least one biologically active agent is selected from the group consisting of peptide, protein, enzyme, small molecule drug, dye, lipid, nucleoside, oligonucleotide, cell, virus, liposome, microparticle, micelle, and combinations thereof.

19. The conjugate of claim 1, wherein the at least one biologically active agent is selected from the group consisting of antibiotic, fungicide, anti-viral agent, anti-inflammatory agent, anti-tumor agent, cardiovascular agent, anti-anxiety agent, hormone, growth factor, steroidal agent, and combinations thereof.

20. The conjugate of claim 1, wherein the biologically active agent comprises a thiol group and is attached to the polymer via reaction with the N-maleimidyl moiety.

21. The conjugate of claim 1, wherein the biologically active agent comprises an amino group and is attached to the polymer via reaction with the first functional group.

22. The conjugate of claim 1, wherein a first biologically active agent is attached to the polymer via reaction with the first functional group and a second biologically active agent is attached to the polymer via reaction with the N-maleimidyl moiety.

23. The conjugate of claim 22, wherein the first biologically active agent comprises an amino group and the first functional group is an active ester.

24. The conjugate of claim 22, wherein the second biologically active agent comprises a thiol group.

25. The conjugate of claim 22, wherein the first and second biologically active agents are proteins or peptides.

26. A polymer conjugate comprising a multi-functional polymer attached to at least one biologically active agent, said polymer having the structure:

wherein;

POLY is poly(ethylene glycol) having an average molecular weight from about 800 Da to about 100,000 Da and a terminal carbon, the terminal carbon being directly bonded to the nitrogen atom of the N-maleimidyl moiety; and

X is selected from the group consisting of —

CO₂H, —

CHO, —

CH(OC₂H₅)₂, N-hydroxysuccinimidyl ester, 1-benzotriazolyl ester, N-hydroxysuccinimidyl carbonate, 1-benzotriazolyl carbonate, and tresylate;

wherein said biologically active agent is attached to the polymer via reaction with the N-maleimidyl moiety or X.

27. The conjugate of claim 26, wherein the at least one biologically active agent is selected from the group consisting of peptide, protein, enzyme, small molecule drug, dye, lipid, nucleoside, oligonucleotide, cell, virus, liposome, microparticle, micelle, and combinations thereof.

28. The conjugate of claim 26, wherein the at least one biologically active agent is selected from the group consisting of antibiotic, fungicide, anti-viral agent, anti-inflammatory agent, anti-tumor agent, cardiovascular agent, anti-anxiety agent, hormone, growth factor, steroidal agent, and combinations thereof.

29. The conjugate of claim 26, wherein the biologically active agent comprises a thiol group and is attached to the polymer via reaction with the N-maleimidyl moiety.

30. The conjugate of claim 26, wherein the biologically active agent comprises an amino group and is attached to the polymer via reaction with X.

31. The conjugate of claim 26, wherein a first biologically active agent is attached to the polymer via reaction with X and a second biologically active agent is attached to the polymer via reaction with the N-maleimidyl moiety.

32. The conjugate of claim 31, wherein the first biologically active agent comprises an amino group and X is N-hydroxysuccinimidyl ester.

33. The conjugate of claim 31, wherein the second biologically active agent comprises a thiol group.

34. The conjugate of claim 31, wherein the first and second biologically active agents are proteins or peptides.

35. A polymer conjugate comprising a branched multi-functional polymer attached to at least one biologically active agent, said branched polymer comprising a central branched core moiety and a plurality of water soluble and non-peptidic polymer backbones attached to said central branched core moiety, each polymer backbone comprising a terminal carbon and having an average molecular weight from about 800 Da to about 100,000 Da, wherein said terminal carbon of each polymer backbone is directly bonded to a N-maleimidyl moiety having the structure:

wherein L is the point of bonding to the terminal carbon of each polymer backbone, and wherein said biologically active agent is attached to the polymer via reaction with the N-maleimidyl moiety.

36. The conjugate of claim 35, wherein each polymer backbone is poly(ethylene glycol) having an average molecular weight from about 6,000 Da to about 80,000 Da.

37. The conjugate of claim 35, wherein the central branched core moiety is a polyol or an amino acid.

38. The conjugate of claim 35, wherein the central branched core moiety is selected from the group consisting of glycerol, glycerol oligomers, pentaerythritol, sorbitol, and lysine.

39. The conjugate of claim 35, wherein the polymer has the structure:

wherein;

POLY is a water-soluble and non-peptidic polymer backbone having a terminal carbon, the terminal carbon being directly bonded to the nitrogen atom of the N-maleimidyl moiety;

R is a central core molecule; and

q is an integer from 2 to about 300.

40. The conjugate of claim 39, wherein R is selected from the group consisting of glycerol, glycerol oligomers, pentaerythritol, sorbitol, and lysine.

41. The conjugate of claim 39, wherein POLY is poly(ethylene glycol) having an average molecular weight from about 6,000 Da to about 80,000 Da.

42. The conjugate of claim 39, wherein q is 4.