Patent EP-3301086-A1 (EP)
Inventor: KUBOYAMA TAKESHI
IPC: A61K47/16
Filed: 2012-11-02
Abstract: The present invention relates to a method for producing a composition containing a complex between a membrane composed of a lipid monolayer (reversed micelle) and a nucleic acid, and a lipid membrane for encapsulating the complex therein, comprising the following steps A to D: 
Step A: preparing a complex between the nucleic acid and a liposome comprising a cationic lipid represented by the formula (I) and/or a cationic lipid other than cationic lipid represented by the formula (I); 
Step B: preparing a dispersion liquid by dispersing the complex in water or ethanol aqueous solution; 
Step C: preparing a solution by dissolving the formula (I) and/or the cationic lipid other than cationic lipid represented by the formula (I) in ethanol or an ethanol aqueous solution; and 
Step D: mixing the dispersion liquid and the solution, and optionally adding water; 
wherein the formula (I) is
     
(wherein: 
R 1  and R 2  are, the same or different, each linear or branched alkyl, alkenyl or alkynyl having 12 to 24 carbon atoms, 
X 1  and X 2  are hydrogen atoms, or are combined together to form a single bond or alkylene, 
X 3  is absent or is alkyl having 1 to 6 carbon atoms, or alkenyl having 3 to 6 carbon atoms, 
when X 3  is absent, 
Y is absent, a and b are 0, L 3  is a single bond, R 3  is alkyl having 1 to 6 carbon atoms, alkenyl having 3 to 6 carbon atoms, or alkyl having 1 to 6 carbon atoms or alkenyl having 3 to 6 carbon atoms substituted with 1 to 3 substituent(s), which is(are), the same or different, amino, monoalkylamino, dialkylamino, trialkylammonio, hydroxy, alkoxy, carbamoyl, monoalkylcarbamoyl, or dialkylcarbamoyl, and L 1  and L 2  are -O-, 
Y is absent, a and b are, the same or different, 0 to 3, and are not 0 at the same time, L 3  is a single bond, R 3  is alkyl having 1 to 6 carbon atoms, alkenyl having 3 to 6 carbon atoms, or alkyl having 1 to 6 carbon atoms or alkenyl having 3 to 6 carbon atoms substituted with 1 to 3 substituent(s), which is(are), the same or different, amino, monoalkylamino, dialkylamino, trialkylammonio, hydroxy, alkoxy, carbamoyl, monoalkylcarbamoyl, or dialkylcarbamoyl, L 1  and L 2  are, the same or different, -O-, -CO-O- or -O-CO-, 
Y is absent, a and b are, the same or different, 0 to 3, L 3  is a single bond, R 3  is a hydrogen atom, and L 1  and L 2  are, the same or different, -O-, -CO-O- or -O-CO-, or 
Y is absent, a and b are, the same or different, 0 to 3, L 3  is -CO- or -CO-O-, R 3  is alkyl having 1 to 6 carbon atoms or alkenyl having 3 to 6 carbon atoms substituted with 1 to 3 substituent(s), which is(are), the same or different, amino, monoalkylamino, dialkylamino, trialkylammonio, hydroxy, alkoxy, carbamoyl, monoalkylcarbamoyl, or dialkylcarbamoyl, wherein at least one of the substituents is amino, monoalkylamino, dialkylamino, or trialkylammonio, and L 1  and L 2  are , the same or different, -O-, -CO-O- or -O-CO-, and 
when X 3  is alkyl having 1 to 6 carbon atoms or alkenyl having 3 to 6 carbon atoms, Y is a pharmaceutically acceptable anion, a and b are, the same or different, 0 to 3, L 3  is a single bond, R 3  is alkyl having 1 to 6 carbon atoms, alkenyl having 3 to 6 carbon atoms, L 1  and L 2  are, the same or different, -O-, -CO-O- or -O-CO-).