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

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.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application claims benefit under 35 U.S.C. 119(e) of U.S. Provisional No. 60 / 305,263, filed Jul. 13, 2001, entitled “NANO HYALURONIC ACID AND METHODS OF MAKING AND USING SAME,” the contents of which is expressly incorporated herein in its entirety by reference. [0002] This application is a continuation-in-part of copending U.S. Ser. No. 09 / 437,277, filed Nov. 11, 1999, entitled “POLYMER GRAFTING BY POLYSACCHARIDE SYNTHASES,” which claims benefit under 35 U.S.C. 119(e) of U.S. Provisional No. 60 / 107,929, filed Nov. 11, 1998, entitled “POLYMER GRAFTING BY POLYSACCHARIDE SYNTHASES,” the contents of both of which are expressly incorporated herein in their entirety by reference. [0003] This application is also a continuation-in-part of copending U.S. Ser. No. 09 / 283,402, filed Apr. 1, 1999, entitled “DNA ENCODING HYALURONAN SYNTHASE FROM PASTEURELLA MULTOCIDA AND METHODS;” which claims benefit under 35 U.S.C. 119(e) of U.S. Provisional ...

AI Technical Summary

Benefits of technology

[0045] The most promising initial target oligosaccharides for inhibition of cancer metastasis are HA chains composed of 10 to 14 sugars. The two current competing state-of-the-art techniques for creating the desired HA-oligosaccharides are extremely limited and will not allow the medical potential of the sugars to be achieved. Small HA molecules are presently made either by: (1) partially depolymerizing costly large polymers with degradative enzymes or with chemical means (e.g. heat, acid, sonification), or (2) highly demanding organic chemistry-based carbohydrate synthesis. The former method is difficult to control, inefficient, costly, and is in a relatively stagnant development stage. For example, the enzyme wants to degrade the polymer to the 4 sugar end stage product, but this sugar is inactive for cancer treatment. The use of acid hydrolysis also removes a fraction of the acetyl groups from the GlcNAc groups thereby introducing a positive charge into an otherwise anionic molecule. The latter

Problems solved by technology

In general, these membrane-bound synthase proteins are difficult to manipulate by typical procedures, and only a few enzymes have been identified after biochemical purification.

Despite this sequence information, the molecular details concerning the three-dimensional native structures, the active sites, and the mechanisms of catalytic action of the polysaccharide syntheses, in general, are very limited or absent.

However, the synthetic control and the reproducibility of large-scale reactions are not always successful.

Additionally, such

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