NUCLEIC ACID DELIVERY USING MODIFIED CHITOSANS

• US 20120295355A1
• Filed: 03/12/2012
• Published: 11/22/2012
• Est. Priority Date: 01/29/2009
• Status: Abandoned Application

• Current Assignee: Ruth Baxter, Shenda Baker, Snezna Rogelj, William P. Wiesmann
• Original Assignee: Ruth Baxter, Shenda Baker, Snezna Rogelj, William P. Wiesmann
• Inventors: Baker, Shenda, Wiesmann, William P., Baxter, Ruth, Rogelj, Snezna
• Application Number:
  US13/417,992
• Publication Number:
  US 20120295355A1
• Time in Patent Office: Days
• US Class Current: 435/455
• CPC Class Codes: A61K 48/0041 the non-active part being polymeric
• CPC Class Codes: C12N 15/88 using microencapsulation, e.g. using amphiphile liposome vesicle

Abstract

The present invention is directed to the delivery of nucleic acids in a non-viral vector to cells by positively charged chitosan derivatives, including but not limited to chitosan-arginine, chitosan-lysine and chitosan-histidine.

Claims

1-195. -195. (canceled)

196. A method of transfecting a cell with a nucleic acid, the method comprising:

providing a cell; and

contacting said cell with a composition comprising the nucleic acid, and a functionalized chitosan of the following formula (I);

197. The method of claim 196, wherein the composition comprises a complex or particle, wherein the complex or particle comprises the chitosan derivative and the nucleic acid.

198. The method of claim 196, wherein 55-90% of R¹ substituents are hydrogen, 4-20% of R¹ substituents are acetyl, 4-30% of R¹ substituents are a group of formula (II).

201. The method of claim 196, wherein the molecular weight of the functionalized chitosan is between about 10,000 and about 1,000,000 Da.

202. The method of claim 196, wherein the functionalized chitosan is soluble in aqueous solution between pH 6 and pH 8.

203. The method of claim 196, wherein the functionalized chitosan is substantially free of other impurities.

204. The method of claim 196, wherein the composition further comprises a lipid or a lipid formulation.

205. The method of claim 196, wherein the nucleic acid comprises a DNA or RNA.

206. The method of claim 196, wherein the nucleic acid comprises a therapeutic gene.

207. The method of claim 196, wherein the nucleic acid comprises a vector.

199-200. -200. (canceled)

208. A kit comprising a functionalized chitosan of formula (I):

209. The kit of claim 208, further comprising a nucleic acid.

210. A pharmaceutical composition comprising a nucleic acid and a functionalized chitosan of formula (I):

211. The composition of claim 210, comprising a complex or particle, wherein the complex or particle comprises the chitosan derivative and the nucleic acid.

212. The composition of claim 210, further comprising a second transfection reagent.

213. The composition of claim 210, wherein the nucleic acid comprises a DNA or RNA.

214. The composition of claim 210, wherein the nucleic acid comprises a therapeutic gene.

215. A reaction mixture comprising a nucleic acid and a functionalized chitosan of formula (I):

216. The reaction mixture of claim 215, wherein the nucleic acid comprises a DNA or RNA.

217-219. -219. (canceled)

220. A chitosan derivative/nucleic acid complex comprising:

a functionalized chitosan of formula (I);

221-223. -223. (canceled)

224. A method of delivering a nucleic acid to a cell, the method comprising:

providing chitosan derivative/nucleic acid complex comprising a functionalized chitosan of formula (I);

225. A method of transfecting a cell with a nucleic acid, the method comprising:

providing a cell;

contacting the cell with a functionalized chitosan of formula (I);