POLYMER NANOPARTICLE COMPOSITION FOR PLASMID DNA DELIVERY, AND PREPARATION METHOD THEREFOR
Disclosed are a pharmaceutical composition containing plasmid DNA, characterized by that it comprises plasmid DNA as active ingredients; a peptide comprising a nuclear localization signal (NLS) sequence or RGD peptide sequence; a cationic compound; and an amphiphilic copolymer, and the plasmid DNA binds to a peptide to form a complex with the cationic compound and the complex is entrapped in the nanoparticle structure of the amphiphilic block copolymer, and a preparation method thereof.
1 . A composition for delivering plasmid DNA, comprising
plasmid DNA as active ingredients;
a peptide comprising a nuclear localization signal (NLS) sequence or RGD peptide sequence;
a cationic compound; and
an amphiphilic block copolymer,
characterized by that the plasmid DNA binds to the peptide to form a complex with the cationic compound by electrostatic interactions, and the complex is entrapped in a nanoparticle structure of the amphiphilic block copolymer.
2 . The composition for delivering plasmid DNA according to claim 1 , wherein the plasmid DNA consists of 30,000 base pairs or more.
3 . The composition for delivering plasmid DNA according to claim 1 , wherein the plasmid DNA is comprised in an amount of 0.001 to 10% by weight based on the weight of the total composition to be finally prepared.
4 . The composition for delivering plasmid DNA according to claim 1 , wherein the peptide is covalently bound to a DNA binding group via a linker.
5 . The composition for delivering plasmid DNA according to claim 4 , wherein the DNA binding group is polyamine.
6 . The composition for delivering plasmid DNA according to claim 5 , wherein the polyamine is spermine.
7 . The composition for delivering plasmid DNA according to claim 1 , wherein the cationic compound is one or more selected from the group consisting of a cationic lipid and a cationic polymer.
8 . The composition for delivering plasmid DNA according to claim 7 , wherein the cationic lipid is a cationic lipid of the following Chemical formula 1:
in the formula,
n, m and 1 are respectively 0 to 12, with a proviso that 1≤n+m+1≤12, and a, b and c are respectively 1 to 6, and R1, R2 and R3 are each independently hydrogen or a saturated and unsaturated hydrocarbon having 11 to 25 carbon atoms, with a proviso that at least one of R1, R2 and R3 is a saturated or unsaturated hydrocarbon having 11 to 25 carbon atoms.
9 . The composition for delivering plasmid DNA according to claim 1 , wherein the cationic compound is used in an amount of 0.01 to 50% by weight based on the total weight of the final composition.
10 . The composition for delivering plasmid DNA according to claim 1 , wherein the ratio of the cationic charge (N) of the cationic compound to the anionic charge (P) of the plasmid DNA (N/P) is 0.1 to 128.
11 . The composition for delivering plasmid DNA according to claim 1 , wherein the amphiphilic block copolymer is an A-B type block copolymer comprising a hydrophilic block (A) and a hydrophobic block (B), and the hydrophilic A block is one or more selected from the group consisting of polyalkylene glycol, polyvinyl alcohol, polyvinylpyrrolidone, polyacrylamide and its derivatives, and the hydrophobic B block is one or more selected from the group consisting of polyester, polyanhydride, polyamino acid, polyorthoester, and polyphosphazene.
12 . The composition for delivering plasmid DNA according to claim 1 , wherein the ratio of (a) the weight of the plasmid DNA and cationic compound complex to (b) the weight of the amphiphilic block copolymer [a/b×100; (plasmid DNA weight+cationic compound weight)/amphiphilic block copolymer weight×100] is 0.001 to 100% by weight.
13 . The composition for delivering plasmid DNA according to claim 11 , wherein the terminal hydroxyl group of the hydrophobic B block is modified with one or more selected from the group consisting of cholesterol, tocopherol and a fatty acid having 10 to 24 carbon atoms.
14 . The composition for delivering plasmid DNA according to claim 1 , wherein the content of the amphiphilic block copolymer is 40 to 99.98% by weight based on the total dry weight of the final composition.
15 . The composition for delivering plasmid DNA according to claim 11 , wherein the weight ratio of the hydrophilic block (A) and the hydrophobic block (B) is respectively 40 to 70% by weight based on the total weight of the amphiphilic block copolymer,
16 . The composition for delivering plasmid DNA according to claim 1 , further comprising a fusogenic lipid.
17 . The composition for delivering plasmid DNA according to claim 4 ,
wherein the linker is one or more selected from the group consisting of SMCC (succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-yl-carboxylate), SMPB (succinimidyl 4-(p-maleimidophenyl)butyrate) and GMBS (N-γ-maleimidobutyryl-oxysuccinimide ester).