GLP-1 gene delivery for the treatment of type 2 diabetes
This patent discloses compositions and methods of use thereof to normalize the blood glucose levels of patients with type 2 diabetes. It relates particularly to a plasmid comprising a chicken β actin promoter and enhancer; a modified GLP-1 (7-37) cDNA (pβGLP1), carrying a furin cleavage site, which is constructed and delivered into a cell for the expression of active GLP-1.
1 . A method of normalizing blood glucose levels of a warm blooded animal having type 2 diabetes, comprising:
providing a transfection formulation comprising a cationic polymeric gene carrier complexed with a selected plasmid consisting essentially of: an expression facilitating sequence derived from chicken β-actin promoter and enhancer; an expression sequence comprising ATG (start codon) followed by a sequence SEQ ID NO:5 (CGTCAACGTCGT) coding for a furin cleavage site (FCS) and a sequence coding for the active form of GLP-1 (7-37) or derivatives thereof that are operably linked to said expression facilitating sequence in a proper charge ratio (positive charge of the copolymer/negative charge of the nucleic acid) that is optimally effective for both in vivo and in vitro transfection; and
administering to the animal to be treated an effective amount of the composition such that the cell internalizes the GLP-1 gene.
2 . The method of claim 1 , wherein the cationic polymeric gene carrier is PAGA.
3 . The method of claim 2 , wherein the weight ratio of DNA to PAGA is preferably within a range of 1:0.82 to 1:2.46.
4 . The method of claim 2 , wherein plasmid comprising an nucleotide sequence represented by the SEQ ID NO:1 or having the nucleotide sequence represented by the SEQ ID NO:2.
5 . The method of claim 2 , wherein the cationic polymeric gene carrier further comprises a targeting moiety.