Site-selective modification of polysaccharides and applications thereof

Patent metadata

Field	Value
Publication number	US-10968286-B2
Application number	US-201816223126-A
Country	US
Kind code	B2
Filing date	Dec 18, 2018
Priority date	Jun 2, 2014
Publication date	Apr 6, 2021
Grant date	Apr 6, 2021

Abstract

The present invention relates to site-selective modification of polysaccharides at their reducing end by conjugation with a single aminoxy-Regioselective Addressable Functionalized Template (RAFT) peptide.

First claim

The invention claimed is:

1. A modified polysaccharide selectively modified at its reducing end by conjugation with a single aminoxy-Regioselective Addressable Functionalized Template (RAFT) peptide, wherein said RAFT peptide is a biologically active linear or cyclic RAFT peptide comprising an arginine-glycine-aspartate (RGD) sequence or an heparin binding peptide (HBP) sequence or a combination thereof, or a nuclear localizing sequence (NLS), said RAFT peptide selected from the sequence GGGGRGDYK(SEQ ID NO: 3), GGGGSPPRRARVTY (SEQ ID NO: 4), GGGGSPPLLALVTY (SEQ ID NO: 5), GGGGSPPRRARVTYK (SEQ ID NO: 6), YDGRGGG-(K—ONH2)-GGGGSPPRRARVTY-ONH2 (SEQ ID NO: 7), and GGGGPKKKRKVED (NLS; SEQ ID NO: 8).

2. The modified polysaccharide of claim 1 , wherein said polysaccharide is selected from chitosan, dextran, heparin, heparan sulfate, keratin sulfate, pectin, starch, hyaluronic acid, sulfated hyaluronan, alginate, alginate sulfate, and chondroitin sulfate.

3. The modified polysaccharide of claim 2 , wherein said polysaccharide is alginate or alginate sulfate.

4. The modified polysaccharide of claim 3 , wherein said RAFT comprises an RGD, said modified polysaccharide selected from alginate-GGGGRGDYK, alginate sulfate-GGGGRGDYK, alginate-YDGRGGG-(K—ONH2)-GGGGSPPRRARVTY-ONH2, and alginate sulfate-YDGRGGG-(K—ONH2)-GGGGSPPRRARVTY-ONH2.

5. The modified polysaccharide of claim 2 , wherein said polysaccharide is a dextran.

6. The modified polysaccharide of claim 5 , wherein said RAFT comprises an RGD, said modified polysaccharide selected from dextran-GGGGRGDYK, and dextran-YDGRGGG-(K—ONH2)-GGGGSPPRRARVTY-ONH2.

7. The modified polysaccharide of claim 2 , wherein said polysaccharide is a chitosan.

8. The modified polysaccharide of claim 7 , wherein said RAFT comprises an RGD, said modified polysaccharide selected from chitosan-GGGGRGDYK, and chitosan-YDGRGGG-(K—ONH2)-GGGGSPPRRARVTY-ONH2.

9. The modified polysaccharide of claim 2 , wherein said polysaccharide is hyaluronic acid or sulfated hyaluronan.

10. The modified polysaccharide of claim 9 , wherein said RAFT comprises an RGD or an NLS, said modified polysaccharide selected from hyaluronic acid-GGGGRGDYK, hyaluronic acid-YDGRGGG-(K—ONH2)-GGGGSPPRRARVTY-ONH2, hyaluronic acid-NLS, sulfated hyaluronan-GGGGRGDYK, sulfated hyaluronan-YDGRGGG-(K—ONH2)-GGGGSPPRRARVTY-ONH2, hyaluronan sulfate-GGGGPKKKRKVED, and sulfated hyaluronan-PEG-GGGGPKKKRKVED.

11. The modified polysaccharide of claim 1 , wherein said RAFT peptide is further labeled.

12. The modified polysaccharide of claim 11 , wherein said label is selected from a dye, a fluorescent dye, biotin, a PEG, or a nuclear localization signal (NLS).

13. The modified polysaccharide of claim 12 , wherein said RAFT is NLS and said label is PEG, said labeled RAFT set forth as H2NO-PEG-GGGGPKKKRKVED.

14. The modified polysaccharide of claim 12 , wherein said dye is a fluorophore label 5-carboxytetramethylrhodamine (TAMRA), or a fluorophore label FITC, or a fluorophore labeled Rhodamine-B, linked to the lysine side chain of said RAFT peptide.

15. The modified polysaccharide of claim 14 , wherein said RAFT is GGGGRGDYK (SEQ ID NO: 3) and said dye is FITC.

16. The modified polysaccharide of claim 14 , wherein said RAFT is GGGGSPPRRARVTYK (SEQ ID NO: 6) and said dye is Rhodamine-B.

17. The modified polysaccharide of claim 12 , wherein said RAFT is NLS and said dye is TAMRA.

18. A process for the preparation of a modified polysaccharide according to claim 1 , comprising conjugation of a polysaccharide with an aminoxy-functionalized peptide in the presence of aniline.

Assignees

• B G Negev Tech And Applications Ltd

Inventors

• Cohen Smadar
• Brik Ashraf

Classifications

• C08B37/0021

  Dextran, i.e. (alpha-1,4)-D-glucan; Derivatives thereof, e.g. Sephadex, i.e. crosslinked dextran · CPC title

• C08B37/0072 Primary

  Hyaluronic acid, i.e. HA or hyaluronan; Derivatives thereof, e.g. crosslinked hyaluronic acid (hylan) or hyaluronates · CPC title

• C07K9/001

  the peptide sequence having less than 12 amino acids and not being part of a ring structure · CPC title

• C08B37/0084

  Guluromannuronans, e.g. alginic acid, i.e. D-mannuronic acid and D-guluronic acid units linked with alternating alpha- and beta-1,4-glycosidic bonds; Derivatives thereof, e.g. alginates · CPC title

• C08B37/00

  Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof (cellulose D21; {microbiological processes C12P}) · CPC title