Claims
- 1. A multi-functional N-maleimidyl polymer derivative 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:
- 2. The polymer of claim 1, wherein said water soluble and non-peptidic polymer backbone has from about 2 to about 300 termini.
- 3. The polymer 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 polymer 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 polymer of claim 1, having the structure:
- 6. The polymer of claim 5, wherein POLY is selected from the group consisting of poly(alkylene glycol), copolymers thereof, terpolymers thereof, and mixtures thereof.
- 7. The polymer 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 polymer of claim 5, wherein X is selected from the group consisting of hydroxyl, protected hydroxyl, active ester, such as N-hydroxysuccinimidyl esters and 1-benzotriazolyl esters, active carbonate, such as N-hydroxysuccinimidyl carbonates and 1-benzotriazolyl carbonates, acetal, aldehyde, aldehyde hydrates, alkenyl, acrylate, methacrylate, acrylamide, active sulfone, amine, 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.
- 9. The polymer of claim 1, wherein the water soluble and non-peptidic polymer backbone has the structure:
- 10. The polymer of claim 9, wherein X is selected from the group consisting of hydroxyl, protected hydroxyl, active ester, active carbonate, acetal, aldehyde, aldehyde hydrates, alkenyl, acrylate, methacrylate, acrylamide, active sulfone, amine, 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.
- 11. The polymer of claim 9, wherein X is selected from the group consisting of —OH, —NH2, —CO2H, —CHO, —CH(OC2H5)2, N-hydroxysuccinimidyl esters, 1-benzotriazolyl esters, N-hydroxysuccinimidyl carbonates, 1-benzotriazolyl carbonates, and tresylate.
- 12. The polymer of claim 1, having the structure:
- 13. The polymer of claim 1, having the structure:
- 14. The polymer of claim 13, wherein R is selected from the group consisting of glycerol, glycerol oligomers, pentaerythritol, sorbitol, and lysine.
- 15. A method of preparing a multi-functional N-maleimidyl polymer derivative, comprising:
providing 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 bonded to an amine group; reacting the polymer backbone with maleic anhydride to form an open ring intermediate; and heating the open ring intermediate in the presence of acetic anhydride and a salt of acetic acid to form a multi-functional N-maleimidyl polymer derivative product.
- 16. The method of claim 15, wherein the water soluble and non-peptidic polymer backbone has from about 2 to about 300 termini.
- 17. The method of claim 15, wherein the water soluble and non-peptidic polymer backbone is selected from the group consisting of poly(alkylene glycol), copolymers thereof, terpolymers thereof, and mixtures thereof.
- 18. The method of claim 15, wherein the 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.
- 19. The method of claim 15, wherein said reacting step comprises reacting maleic anhydride with X-PEG-NH2, wherein PEG is poly(ethylene glycol) and X is a functional group, to form an open ring intermediate having the following structure:
- 20. The method of claim 19, wherein X is selected from the group consisting of hydroxyl, protected hydroxyl, acetal, alkenyl, amine, protected amine, protected hydrazide, protected thiol, carboxylic acid, protected carboxylic acid, maleimide, dithiopyridine, and vinylpyridine.
- 21. The method of claim 15, further comprising purifying the open ring intermediate by precipitation prior to said heating step.
- 22. The method of claim 15, wherein said heating step comprises heating the open ring intermediate at a temperature of about 50° C. to about 140° C. for about 0.2 hours to about 5 hours.
- 23. The method of claim 15, further comprising purifying the multi-functional N-maleimidyl polymer derivative product.
- 24. The method of claim 23, wherein said purifying step comprises:
passing the N-maleimidyl polymer derivative product through an ion exchange column; collecting an eluent containing the N-maleimidyl polymer derivative product from the column; and precipitating the N-maleimidyl polymer derivative product by contacting the product with a solvent.
- 25. The method of claim 24, wherein the solvent is selected from the group consisting of ethyl ether, isopropanol, and mixtures thereof
- 26. The method of claim 15, further comprising the step of reacting the N-maleimidyl polymer derivative product with a biologically active agent to form a biologically active polymer conjugate.
- 27. The method of claim 26, wherein the biologically active agent is selected from the group consisting of peptides, proteins, enzymes, small molecule drugs, dyes, lipids, nucleosides, oligonucleotides, cells, viruses, liposomes, microparticles and micelles.
- 28. A method of preparing a N-maleimidyl polymer derivative, comprising
providing 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 terminal amine group; and reacting the polymer backbone with an N-alkoxycarbonylmaleimide to form a N-maleimidyl polymer derivative product.
- 29. The method of claim 32, wherein the water-soluble and non-peptidic polymer backbone has the formula X-PEG-NH2, wherein PEG is poly(ethylene glycol) and X is a capping group.
- 30. The method of claim 29, wherein X is selected from the group consisting of alkoxy, hydroxyl, protected hydroxyl, acetal, alkenyl, amine, protected amine, protected hydrazide, protected thiol, carboxylic acid, protected carboxylic acid, maleimide, dithiopyridine, and vinylpyridine.
- 31. The method of claim 28, further comprising purifying the N-maleimidyl polymer derivative product.
- 32. The method of claim 28, wherein the water soluble and non-peptidic polymer backbone has from about 2 to about 300 termini.
- 33. The method of claim 28, wherein the water soluble and non-peptidic polymer backbone is selected from the group consisting of poly(alkylene glycol), copolymers thereof, terpolymers thereof, and mixtures thereof.
- 34. The method of claim 28, wherein the 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.
- 35. The method of claim 28, further comprising the step of reacting the N-maleimidyl polymer derivative product with a biologically active agent to form a biologically active polymer derivative.
- 36. The method of claim 35, wherein the biologically active agent is selected from the group consisting of peptides, proteins, enzymes, small molecule drugs, dyes, lipids, nucleosides, oligonucleotides, cells, viruses, liposomes, microparticles and micelles.
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional Application No. 60/183,833, filed on Feb. 22, 2000, which is incorporated by reference herein in its entirety.
Provisional Applications (1)
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Number |
Date |
Country |
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60183833 |
Feb 2000 |
US |