Efficient nucleic acid encapsulation into medium sized...

A - Human Necessities – 61 – K

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A61K 9/127 (2006.01) A01N 43/04 (2006.01) C12N 15/88 (2006.01)

Patent

CA 2472462

A method for preparing liposomes containing at least one nucleic acid encapsulated therein comprising the following steps: (A) mixing a gel or a liquid containing gel particles with aqueous medium Z1 to directly form the liposomes containing the at least one nucleic acid encapsulated therein; (B) (i) mixing a gel or a liquid containing gel particles with aqueous medium Z1 to form a curd or curdy substance; and (ii) mixing the curd or curdy substance with aqueous medium Z2 to directly form the liposomes containing the at least one nucleic acid encapsulated therein; or (C) (i) cooling a gel or a liquid containing gel particles to form a waxy substance; and (ii) mixing the waxy substance with aqueous medium Z1 to directly form the liposomes containing the at least one nucleic acid encapsulated therein; wherein said gel or liquid containing gel particles comprises at least one liposome-forming lipid, at least one fusogenic lipid, a water-miscible organic solvent and the at least one nucleic acid; wherein an amount of the at least one fusogenic lipid is at least 20% by weight of a lipid content of the gel or the liquid containing gel particles; and wherein the aqueous media Z1 and Z2 are the same or different.

L'invention concerne un procédé de préparation de liposomes contenant au moins un acide nucléique encapsulé et consistant à: (A) mélanger un gel ou un liquide contenant des particules de gel avec un milieu aqueux Z1 afin d'obtenir directement les liposomes contenant au moins un acide nucléique encapsulé; (B) (i) mélanger un liquide contenant des particules de gel avec un milieu aqueux Z1 afin d'obtenir un caillé ou une substance avec aspect de caillé; et (ii) mélanger le caillé ou la substance avec aspect de caillé avec un milieu aqueux Z2 afin d'obtenir directement les liposomes contenant au moins un acide nucléique encapsulé ou (C) (i) refroidir un gel ou un liquide contentant des particules de gel afin d'obtenir une substance cireuse; et (ii) mélanger la substance cireuse avec le milieu aqueux Z1 afin d'obtenir directement les liposomes contenant au moins un acide nucléique encapsulé; ledit gel ou liquide contenant les particules de gel renfermant au moins un lipide de formation de liposome, au moins un lipide fusogène, un solvant organique hydrosoluble et au moins un acide nucléique; la quantité d'au moins un lipide fusogène étant d'au moins 20 % en poids de la teneur lipidique du gel ou du liquide contentant les particules de gel; et les milieux aqueux Z1 et Z2 étant identiques ou différents.

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