Natural, chimeric and hybrid glycosaminoglycan polymers and methods of making and using same

• US 20060188966A1
• Filed: 07/15/2002
• Published: 08/24/2006
• Est. Priority Date: 04/02/1998
• Status: Abandoned Application

• Current Assignee: The Board of Regents of the University of Colorado (University of Colorado System)
• Original Assignee: The Board of Regents of the University of Colorado (University of Colorado System)
• Inventors: DeAngelis, Paul L.
• Application Number:
  US10/195,908
• Publication Number:
  US 20060188966A1
• Time in Patent Office: Days
• US Class Current: 435/85
• CPC Class Codes: A61K 47/36 Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
• CPC Class Codes: A61K 9/006 Oral mucosa, e.g. mucoadhesive forms, sublingual droplets; Buccal patches or films; Buccal sprays
• CPC Class Codes: A61L 24/08 Polysaccharides A61L24/043 takes precedence
• CPC Class Codes: A61L 29/085 Macromolecular materials
• CPC Class Codes: C08B 37/0063 Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
• CPC Class Codes: C08L 5/08 Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof
• CPC Class Codes: C12N 9/1048 Glycosyltransferases (2.4)
• CPC Class Codes: C12N 9/1051 Hexosyltransferases (2.4.1)
• CPC Class Codes: C12P 19/26 Preparation of nitrogen-containing carbohydrates
• CPC Class Codes: C12P 19/28 N-glycosides

Abstract

The present invention relates to methodology for polymer grafting by a polysaccharide synthase and, more particularly, polymer grafting using the hyaluronate or chondroitin or heparin/heparosan synthases from Pasteurella multocida, in order to create a variety of glycosaminoglycan oligosaccharides having a natural or chimeric or hybrid sugar structure.

Claims

1-172. -172. (canceled)

173. A method for producing a chimeric or hybrid glycosaminoglycan having a non-natural structure, comprising the steps of:

providing a functional acceptor, wherein the functional acceptor has at least two sugar units selected from the group consisting of uronic acid and hexosamine;

providing at least one glycosaminoglycan transferase, wherein the glycosaminoglycan transferase is selected from the group consisting of pmHAS, pmCS, pmHS, PglA, and mutants thereof; and

providing at least one of UDP-GlcUA, UDP-GlcNAc, UDP-Glc and UDP-GalNAc such that the at least one glycosaminoglycan transferase elongates the functional acceptor in a manner so as to provide a chimeric or hybrid glycosaminoglycan having a non-natural structure.

174. The method of claim 173, wherein the functional acceptor is a plurality of functional acceptors immobilized on a substrate.

175. The method of claim 173, wherein the functional acceptor is a plurality of functional acceptors in a liquid phase.

176. A chimeric or hybrid glycosaminoglycan having a non-natural structure produced according to the methodology of claim 173, wherein the chimeric or hybrid glycosaminoglycan having a non-natural structure is composed of from about 1 to about 50 sugars.

177. A library of natural glycosaminoglycan polymers produced according to the process comprising the steps of:

treating at least one functional acceptor with at least one glycosaminoglycan transferase in the presence of at least one UDP-sugar selected from the group consisting of UDP-GlcNAc, UDP-GlcUA, UDP-GalNAc, and UDP-GlcN.

178. The product made according to the process of claim 177, wherein the functional acceptor is immobilized on a microtiter plate.

179. The product made according to the process of claim 177, wherein the functional acceptor is immobilized on a microarray slide.

180. The product made according to the process of claim 177, wherein the functional acceptor is in a liquid phase.

181. The product made according to the process of claim 180, wherein the functional acceptor is in the liquid phase and is encoded with a distinct tag capable of decoding structure.

182-187. -187. (canceled)