COMPOSITIONS AND METHODS FOR EFFICACIOUS AND SAFE DELIVERY OF siRNA USING SPECIFIC CHITOSAN-BASED NANOCOMPLEXES

Abstract

There is disclosed a composition and a method for the efficient delivery of a therapeutic RNAi-inducing nucleic acid to cells both in vitro and in vivo through specific formulations of a non viral delivery system using chitosans. Particularly, the composition contains a nucleic acid and a specific chitosan that has the following physico-chemical properties: a number-average molecular weight between 5 kDa and 200 kDa, a degree of deacetylation between 80% and 95% and a chitosan amine to nucleic acid phosphate ratio below 20.

What is claimed is: 
     
         1 . A composition for inhibiting a gene expression in vivo comprising chitosan and an RNAi-inducing nucleic acid sequence against said gene wherein the chitosan has a molecular weight (Mn) of 5 kDa to 200 kDa, a degree of deacetylation (DDA) of 80% to 95%, and wherein the chitosan amine to nucleic acid phosphate ratio (N:P) is below 20. 
     
     
         2 - 6 . (canceled) 
     
     
         7 . The composition of  claim 1 , wherein the RNAi-inducing nucleic acid sequence is a double stranded linear deoxyribonucleic acid or ribonucleic acid sequence between 10 to 50 nucleotides. 
     
     
         8 . The composition of  claim 1 , wherein the RNAi-inducing nucleic acid sequence is a hairpin structure of deoxyribonucleic or ribonucleic acid sequence. 
     
     
         9 . The composition of  claim 1 , wherein the RNAi-inducing nucleic acid sequence is chemically modified either on the sugar backbone, phosphate backbone and/or the nucleotide base ring. 
     
     
         10 . The composition of  claim 1 , wherein the RNAi-inducing nucleic acid sequence is a short interfering RNA, a short hairpin RNA or an RNAi-inducing vector. 
     
     
         11 . The composition of  claim 1 , wherein the RNAi-inducing nucleic acid sequence targets a gene involved in the pathogenesis of type II diabetes, atherosclerosis, cancer, in tumor development, in metastasis, or in induction of chemoresistance. 
     
     
         12 . (canceled) 
     
     
         13 . The composition of  claim 1 , wherein the RNAi-inducing nucleic acid sequence targets a glycoregulating protein, an incretin degrading enzyme, an atherogenic protein, a helicase protein, an RNA helicase, P68, DDX5, DDX32, DDX1, Akt, PKB, a member of the ABC transporters, MDR1, MRP, a member of the RAS family of proteins, SRC, HER2, EGFR, Abl, or Raf. 
     
     
         14 . (canceled) 
     
     
         15 . The composition of  claim 13 , wherein the incretin degrading enzyme is dipeptydilpeptidase-IV (DPP-IV). 
     
     
         16 . (canceled) 
     
     
         17 . The composition of  claim 13 , wherein the atherogenic protein is Apolipoprotein B (ApoB), Apolipoprotein E (ApoE), Apolipoprotein B 100 (ApoB 100), Apolipoprotein B 48 (ApoB 48), Neutrophil gelatinase-associated lipocalin (NGAL), Matrix metalloproteinase-9 (MMP-9), or Cholesteryl ester transfer protein (CETP). 
     
     
         18 . (canceled) 
     
     
         19 . The composition of  claim 13 , wherein the helicase protein is a member of the RecQ family of helicases. 
     
     
         20 . The composition of  claim 13 , wherein the helicase protein is RecQL1 DNA helicase. 
     
     
         21 - 22 . (canceled) 
     
     
         23 . The method of  claim 55 , for treating diabetes mellitus related conditions wherein said diabetes mellitus related conditions are insulin-dependent diabetes mellitus (type I diabetes), noninsulin-dependent diabetes mellitus (type II diabetes), insulin resistance, hyperinsulinemia, diabetes-induced hypertension, obesity, damage to blood vessels, damage to eyes, damage to kidneys, damage to nerves, damage to autonomic nervous system, damage to skin, damage to connective tissue, and damage to immune system. 
     
     
         24 . The method of  claim 55 , for treating atherosclerosis related conditions wherein said atherosclerosis related conditions are cardiovascular diseases. 
     
     
         25 . The method of  claim 24 , wherein the cardiovascular diseases are coronary heart diseases, acute coronary syndromes or angina pectoris. 
     
     
         26 - 54 . (canceled) 
     
     
         55 . A method for treating diabetes mellitus, atherosclerosis or cancer in a patient comprising administering to said patient an effective amount of the composition of  claim 1 . 
     
     
         56 - 75 . (canceled) 
     
     
         76 . The method of  claim 55 , wherein said composition reduces ApoB plasma levels, increases GLP-1 bioavailability, increases the control of glucose metabolism in said patient, reduces the cholesterol level in a patient, reduces the low-density lipoprotein level in said patient, and/or further reduces the weight gain in said patient. 
     
     
         77 - 83 . (canceled) 
     
     
         84 . The method of  claim 55 , said composition further comprising insulin, a glucosidase inhibitor, a sulfonylurea, a DPP-IV inhibitor or a hypoglycemic compound. 
     
     
         85 - 86 . (canceled) 
     
     
         87 . The method of  claim 84 , wherein said hypoglycemic compound is metformin, acarbose, acetohexamide, glimepiride, tolazamide, glipizide, glyburide, tolbutamide, chlorpropamide, thiazolidinediones, alpha glucosidase inhibitors, biguanindine derivatives, troglitazone, or a mixture thereof. 
     
     
         88 . The method of  claim 84 , wherein said sulfonylurea is tolbutanide, tolazamide, glisoxepide, glimipeide or glibomuride. 
     
     
         89 . The method of  claim 84 , wherein said DPP-IV inhibitor is sitagliptin, vildagliptin or saxagliptin. 
     
     
         90 - 96 . (canceled)