Targeted glycosaminoglycan polymers by polymer grafting and methods of making and using same
First Claim
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1. A method for enzymatically producing glycosaminoglycan polymers having unnatural structures, the method comprising the steps of:
- providing at least one functional acceptor, wherein the functional acceptor has at least one sugar unit selected from the group consisting of uronic acid, hexosamine and structural variants or derivatives thereof;
providing at least one recombinant glycosaminoglycan transferase having an empty acceptor site and being capable of elongating the at least one functional acceptor to form extended glycosaminoglycan-like molecules;
providing at least one UDP-sugar analog, wherein the at least one UDP-sugar analog is not found in mammals in a native state; and
wherein the at least one recombinant glycosaminoglycan transferase elongates the at least one functional acceptor to provide glycosaminoglycan polymers wherein the glycosaminoglycan polymers have an unnatural structure.
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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, in order to create a variety of glycosaminoglycan oligosaccharides having a natural or chimeric or hybrid sugar structure with a targeted size that are substantially monodisperse in size. The present invention also relates to methodology for polymer grafting by a polysachharide synthase to form glycosaminoglycan polymers having an unnatural structure.
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Citations
25 Claims
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1. A method for enzymatically producing glycosaminoglycan polymers having unnatural structures, the method comprising the steps of:
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providing at least one functional acceptor, wherein the functional acceptor has at least one sugar unit selected from the group consisting of uronic acid, hexosamine and structural variants or derivatives thereof;
providing at least one recombinant glycosaminoglycan transferase having an empty acceptor site and being capable of elongating the at least one functional acceptor to form extended glycosaminoglycan-like molecules;
providing at least one UDP-sugar analog, wherein the at least one UDP-sugar analog is not found in mammals in a native state; and
wherein the at least one recombinant glycosaminoglycan transferase elongates the at least one functional acceptor to provide glycosaminoglycan polymers wherein the glycosaminoglycan polymers have an unnatural structure. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9)
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10. A method for enzymatically producing glycosaminoglycan polymers having unnatural structures, the method comprising the steps of:
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providing at least one functional acceptor, wherein the functional acceptor has at least one sugar unit selected from the group consisting of uronic acid, hexosamine and structural variants or derivatives thereof;
providing at least one recombinant glycosaminoglycan transferase having an empty acceptor site and being capable of elongating the at least one functional acceptor to form extended glycosaminoglycan-like molecules; and
providing at least one UDP-sugar selected from the group consisting of UDP-GlcUA, UDP-GlcNAc, UDP-Glc, UDP-GalNAc, UDP-GlcN, UDP-GalN and structural variants or derivatives thereof; and
providing at least one UDP-sugar analog, wherein the at least one UDP-sugar analog that is not found in mammals in a native state; and
wherein the at least one recombinant glycosaminoglycan transferase elongates the at least one functional acceptor to provide glycosaminoglycan polymers wherein the glycosaminoglycan polymers have an unnatural structure. - View Dependent Claims (11, 12, 13, 14, 15, 16, 17, 18)
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- 19. A recombinantly produced, isolated glycosaminoglycan polymer having an unnatural structure, wherein the glycosaminoglycan polymer comprises at least one sugar analog that is not found in mammals in a native state.
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23. A recombinantly produced, isolated glycosaminoglycan polymer having an unnatural structure, comprising:
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a glycosaminoglycan selected from the group consisting of an HA oligosaccharide, an HA polymer, a chondroitin oligosaccharide, a chondroitin polymer, a chondroitin sulfate polymer, a heparosan oligosaccharide, a heparin polymer, a heparin polymer, a heparosan polymer, and combinations thereof; and
at least one sugar analog that is not found in mammals in a native state, wherein the at least one sugar analog is selected from the group consisting of UDP-GlcN, UDP-GlcNAcUA, UDP-GlcNAcNAc, UDP-GlcdiNAcUA, UDP-GlcN[TFA], UDP-GlcNBut, U DP-GlcNPro, UDP-6-F-6-deoxyGlcNAc, UDP-2-F-2-deoxyGlcUA, and combinations thereof. - View Dependent Claims (24)
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25. A method for doing business, comprising the steps of:
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providing at least one functional acceptor, wherein the functional acceptor has at least one sugar unit selected from the group consisting of uronic acid, hexosamine and structural variants or derivatives thereof;
providing at least one recombinant glycosaminoglycan transferase having an empty acceptor site and being capable of elongating the at least one functional acceptor to form extended glycosaminoglycan-like molecules;
providing at least one UDP-sugar analog, wherein the at least one UDP-sugar analog is not found in mammals in a native state;
wherein the at least one recombinant glycosaminoglycan transferase elongates the at least one functional acceptor to provide glycosaminoglycan polymers wherein the glycosaminoglycan polymers have an unnatural structure; and
selling and delivering the glycosaminoglycan polymers having an unnatural structure to a customer.
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Specification