Non-viral nanoparticle-based delivery system

Abstract

The present invention concerns a polymeric material for the production of a non-viral nanoparticle. The polymeric material comprises (i) a hydrophilic linear polymer having a first end and a second end, (iii) a cross-linkable cationic polymer covalently bonded to the first end of the hydrophilic linear polymer, and (iii) at least one targeting/penetrating peptide covalently associated to the second end of the hydrophilic linear polymer. Also disclosed herein are nanoparticles produced with these polymeric material, processes for making the polymeric material and the nanoparticles as well as use of the nanoparticles.

1  to  45 . (canceled) 
     
     
         46 . A method of treating a Polo-like kinase-1 (Plk1)-expressing cancer in an individual in need thereof by targeted delivery of a Plk1-targeting siRNA, said method comprising:
 (a) providing a non-viral nanoparticle having and outer layer and an internal core comprising the Plk1-targeting siRNA; and   (b) administering the non-viral nanoparticle to the individual,   wherein the non-viral nanoparticle comprises:   a plurality of polymers, wherein each polymer comprises:   (i) a linear polyethylene glycol polymer (PEG) having a first end and a second end,   (ii) a cross-linkable cationic chitosan polymer covalently associated to the first end of the linear polyethylene glycol polymer via an ester linkage to a monomer of the chitosan polymer,   wherein the cationic chitosan polymer has:
 (1) an average molecular weight of between about 10 and 200 kDa; and 
 (2) a degree of deacetylation between about 70% and 90%; and 
   (iii) at least one targeting/penetrating peptide covalently associated to the second end of the linear polyethylene glycol polymer,   wherein the at least one targeting/penetrating peptide comprises a TAT peptide;   and wherein the plurality of cationic chitosan polymers are cross linked to each to form the outer layer of the nanoparticle to generate the internal core; and the Plk-targeting siRNA is in the internal core, and   the non-viral nanoparticle has an average diameter of between about 5 to 300 nm, or between about 100 nm to 200 nm.   
     
     
         47 . The method of  claim 46 , wherein the non-viral nanoparticle is formulated for intranasal administration and the administration is intranasal. 
     
     
         48 . The method of  claim 46 , wherein the cancer is a brain cancer or a colon cancer. 
     
     
         49 . The method of  claim 46 , wherein the non-viral nanoparticle is formulated for intravenous administration and the administration is intravenous. 
     
     
         50 . The method of  claim 46 , wherein the average molecular weight of the cationic chitosan polymer is from about 50 to about 200 kDa. 
     
     
         51 . The method of  claim 46 , wherein the degree of deactylation of the cationic chitosan polymer is about 80%. 
     
     
         52 . The method of  claim 46 , wherein the non-viral nanoparticle has an average diameter of less than 20 nm. 
     
     
         53 . The method of  claim 46 , wherein the non-viral nanoparticle is formulated for intratumoral administration and the administration is intratumoral. 
     
     
         54 . A non-viral nanoparticle comprising a polymeric material comprising:
 (i) a linear polyethylene glycol polymer having a first end and a second end,   (ii) a cross-linkable cationic chitosan polymer covalently associated to the first end of the linear polyethylene glycol polymer, and   (iii) at least one targeting/penetrating peptide covalently associated to the second end of the linear polyethylene glycol polymer.   
     
     
         55 . The non-viral nanoparticle of  claim 54 , wherein the cationic polymer is cross-linked so as to form an internal core; and an anionic agent entrapped in the internal core. 
     
     
         56 . A polymeric delivery system for nucleic acids and/or anticancer drugs,
 wherein the polymeric delivery system comprises nanoparticles comprising: chitosan, polyethylene glycol and a peptide sequence,   wherein the peptide sequence comprises a cell penetrating peptide, a protein transduction domain, and a cell surface receptor targeting moiety,   wherein the polymeric delivery system is made by a method comprising:
 a. Reaction of amine groups of chitosan with phthalic anhydride as a protection step; 
 b. Activation of monomethoxy polyethylene glycol (mPEG-OH) with succinic anhydride to form mPEG-COOH; 
 c. Activation of mPEG-COOH with thionyl chloride to form mPEG-COCl to form an activated PEG; 
 d. Reaction of hydroxyl groups of chitosan to the activated mPEG-COCl to form PEGylated chitosan; 
 e. Reaction of monomethoxy group of PEGylated chitosan with aluminium chloride in ethane thiol to convert methoxy to hydroxyl group; 
 f. Reaction of hydroxyl terminated PEGylated chitosan with succinic anhydride to form an activated Chitosan-PEG-COOH; 
 g. Reaction of activated Chitosan-PEG-COOH with amine terminated peptide (TAT/MGF/CP15) using EDC and DMAP as a catalyst to form Chitosan-PEG-Peptide; and 
 h. Reaction of Chitosan-PEG-Peptide with hydrazine monohydrate to deprotect amine groups of chitosan.