Dually derivatized chitosan nanoparticles and methods of making and using the same for gene transfer in vivo

• US 10,456,481 B2
• Filed: 08/10/2018
• Issued: 10/29/2019
• Est. Priority Date: 09/25/2013
• Status: Active Grant

• Current Assignee: enGene, Inc.
• Original Assignee: enGene, Inc.
• Inventors: Gao, Jun, Hsu, Eric, Cheung, Anthony
• Primary Examiner(s): Burkhart, Michael D
• Application Number:
  US16/101,327
• Publication Number:
  US 20180344875A1
• Time in Patent Office: 445 Days
• Field of Search: None
• CPC Class Codes: A61K 48/005 characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
• CPC Class Codes: A61K 9/5161 Polysaccharides, e.g. alginate, chitosan, cellulose derivatives; Cyclodextrin
• 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: C12N 15/88 using microencapsulation, e.g. using amphiphile liposome vesicle

Abstract

Provided herein is chitosan-derivative nanoparticle comprising chitosan functionalized with a cationic amino acid and a hydrophilic polyol; and methods of making and using same, e.g., for gene delivery in vivo.

Claims

1. A method of treating diabetes in a patient in need thereof, the method comprising administering a therapeutically effective amount of a therapeutic nucleic acid encoding insulin, a glucagon antagonist, GLP-1 or leptin to a target tissue in the patient, wherein said administering comprises contacting said target tissue with a dually derivatized (DD) chitosan nucleic acid polyplex, said DD chitosan nucleic acid polyplex comprising a chitosan-derivative nanoparticle and said therapeutic nucleic acid, wherein said chitosan-derivative nanoparticle comprises chitosan functionalized with arginine (Arg) and a hydrophilic polyol (HP) of Formula VII:

4. The method of claim 1, 2, or 3, wherein the hydrophilic polyol has a carboxyl group.

5. The method of claim 1, 2, or 3, wherein the hydrophilic polyol is a saccharide selected from the group consisting of glyceraldehyde, threose, erythrose, ribose, arabinose, xylose, lyxose, allose, glucose, altrose, mannose, gulose, idose, galactose, and talose.

6. The method of claim 1, 2, or 3, wherein the nanoparticle has an Arg final functionalization degree:

HP final functionalization degree ratio of between about 1;

1 to about 10;

1.

7. The method of claim 1, 2, or 3, wherein said polyplexes comprise chitosan molecules having an average molecular weight of less than 110 kDa before functionalization.

8. The method of claim 1, 2, or 3, wherein the amine to phosphate ratio of said DD-chitosan nucleic acid polyplex is between 2 to 100.

9. The method of claim 6, wherein the nanoparticle has an Arg final functionalization degree:

HP final functionalization degree ratio of between about 3;

1 to about 7;

1.

10. The method of claim 8, wherein the amine to phosphate ratio of said DD-chitosan nucleic acid polyplex is between 2 to 50, 2 to 30, or 2 to 15.

2. A method of treating inflammatory bowel disease in a patient in need thereof, the method comprising administering a therapeutically effective amount of a therapeutic nucleic acid encoding IL-10, a TNFα

antagonist, or an IL-17 antagonist to a target tissue in the patient, wherein said administering comprises contacting said target tissue with a dually derivatized (DD) chitosan nucleic acid polyplex, said DD chitosan nucleic acid polyplex comprising a chitosan-derivative nanoparticle and said therapeutic nucleic acid, wherein said chitosan-derivative nanoparticle comprises chitosan functionalized with arginine (Arg) and a hydrophilic polyol (HP) of Formula VII;

3. A method of treating of obesity in a patient in need thereof, the method comprising administering a therapeutically effective amount of a therapeutic nucleic acid encoding leptin, cholecystokinin, PYY or GLP-1 to a target tissue in the patient, wherein said administering comprises contacting said target tissue with a dually derivatized (DD) chitosan nucleic acid polyplex, said DD chitosan nucleic acid polyplex comprising a chitosan-derivative nanoparticle and said therapeutic nucleic acid, wherein said chitosan-derivative nanoparticle comprises chitosan functionalized with arginine (Arg) and a hydrophilic polyol (HP) of Formula VII: