Dually derivatized chitosan nanoparticles and methods of making and using the same

• US 9,623,112 B2
• Filed: 03/15/2013
• Issued: 04/18/2017
• Est. Priority Date: 03/21/2012
• Status: Active Grant

• Current Assignee: enGene, Inc.
• Original Assignee: enGene, Inc.
• Inventors: Gao, Jun, Hsu, Eric, Cheung, Anthony
• Primary Examiner(s): MCDONALD, JENNIFER SUE PITRAK
• Application Number:
  US14/386,321
• Publication Number:
  US 20150051265A1
• Time in Patent Office: 1,495 Days
• Field of Search: None
• CPC Class Codes: A61K 47/61 the organic macromolecular compound being a polysaccharide or a derivative thereof
• CPC Class Codes: A61K 47/6939 the polymer being a polysaccharide, e.g. starch, chitosan, chitin, cellulose or pectin
• CPC Class Codes: A61K 9/5161 Polysaccharides, e.g. alginate, chitosan, cellulose derivatives; Cyclodextrin
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C07H 13/04 having the esterifying carboxyl radicals attached to acyclic carbon atoms
• CPC Class Codes: C07H 5/06 Aminosugars
• CPC Class Codes: C08B 37/003 Chitin, i.e. 2-acetamido-2-deoxy-(beta-1,4)-D-glucan or N-acetyl-beta-1,4-D-glucosamine; Chitosan, i.e. deacetylated product of chitin or (beta-1,4)-D-glucosamine; Derivatives thereof
• CPC Class Codes: C08L 5/08 Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2320/32 Special delivery means, e.g. tissue-specific

Abstract

Provided herein is chitosan dually derivatized with arginine and gluconic acid; and methods of making and using the same, e.g., for gene delivery in vivo.

Claims

1. A chitosan-derivative nanoparticle comprising chitosan coupled with gluconic acid and arginine.

2. The nanoparticle according to claim 1, wherein said nanoparticle comprises gluconic acid at an initial concentration of about 5% to about 60%.

3. The nanoparticle according to claim 2, wherein said nanoparticle comprises gluconic acid at an initial concentration of about 8% to about 30%.

4. The nanoparticle according to claim 3, wherein said nanoparticle comprises gluconic acid at a final functionalization of about 3% to about 10%.

5. The nanoparticle according to claim 4, wherein said nanoparticle comprises gluconic acid at a final functionalization of about 5%.

6. The nanoparticle according to claim 1, wherein said nanoparticle comprises arginine at a concentration of about 10% to about 55%.

7. A dually derivatized (DD) chitosan nucleic acid polyplex comprising the nanoparticle according to any one of claims 1-6.

8. The DD-chitosan nucleic acid polyplex according to claim 7, wherein said nucleic acid is DNA or RNA.

9. The DD-chitosan nucleic acid polyplex according to claim 7, wherein the amine to phosphate ratio of said DD-chitosan nucleic acid polyplex is between 2 to 100.

10. The DD-chitosan nucleic acid polyplex according to claim 9, wherein said amine to phosphate ratio of said DD-chitosan nucleic acid polyplex is between 2 to 50.

11. The DD-chitosan nucleic acid polyplex according to claim 10, wherein said amine to phosphate ratio of said DD-chitosan nucleic acid polyplex is between 2 to 30.

12. The DD-chitosan nucleic acid polyplex according to claim 11, wherein said amine to phosphate ratio of said DD-chitosan nucleic acid polyplex is between 2 to 15.

13. A method of delivering a nucleic acid molecule to a cell comprising contacting said cell with a DD-chitosan nucleic acid polyplex according to claim 7.

14. The method of claim 13, wherein said cell is in vivo.

15. The DD-chitosan nucleic acid polyplex according to claim 7, wherein said DD-chitosan nucleic acid polyplex has a combined degree of functionalization with said arginine and said gluconic acid of 1-60%.

16. The DD-chitosan nucleic acid polyplex according to claim 7, wherein said DD-chitosan nucleic acid polyplex has a combined degree of functionalization with said arginine and said gluconic acid of 1-30%.

17. The DD-chitosan nucleic acid polyplex according to claim 7, wherein said nucleic acid is selected from the group consisting of peptide nucleic acid (PNA), phosphorodiamidate morpholino oligo (PMO), locked nucleic acid (LNA), glycol nucleic acid (GNA), and threose nucleic acid (TNA).

18. The DD-chitosan nucleic acid polyplex according to claim 8, wherein said RNA is selected from the group consisting of antisense RNA, siRNA, short hairpin RNA, micro RNA, and enzymatic RNA.

19. The DD-chitosan nucleic acid polyplex according to claim 7, wherein said chitosan molecules have an average molecular weight of less than 110 kDa.

20. The DD-chitosan nucleic acid polyplex according to claim 7, wherein said DD-chitosan nucleic acid polyplex has an average polydispersity index (PDI) selected from the group consisting of less than 0.5, less than 0.4, less than 0.3, and less than 0.25.

21. The DD-chitosan nucleic acid polyplex according to claim 7, wherein said DD-chitosan nucleic acid polyplex has a molar ratio of said arginine to said gluconic acid of between 100:

1 and 1;

100.

22. The DD-chitosan nucleic acid polyplex according to claim 21, wherein said molar ratio of said arginine to said gluconic acid is between 50:

1 and 1;

50.

23. The DD-chitosan nucleic acid polyplex according to claim 21, wherein said molar ratio of said arginine to said gluconic acid is between 10:

1 and 1;

10.

24. The DD-chitosan nucleic acid polyplex according to claim 21, wherein said molar ratio of said arginine to said gluconic acid is between 5:

1 and 1;

5.

25. The DD-chitosan nucleic acid polyplex according to claim 21, wherein said molar ratio of said arginine to said gluconic acid is between 2:

1 and 1;

2.

26. A pharmaceutical composition comprising said DD-chitosan nucleic acid polyplex of claim 7.