Lipid nanoparticles for transfection and related methods

Abstract

Transfection reagent composition, lipid nanoparticles prepared from the transfection reagent composition, kits that include the transfection reagent composition, and methods for making and using lipid nanoparticles prepared from the transfection reagent composition. Lipids when dispersed in aqueous media readily form liposomes, such as unilamellar vesicles and multilamellar vesicles. Liposomes have been used successfully to encapsulate and deliver a wide range of chemicals including nucleic acids, proteins and, small molecule drugs, to cells.

1 . A transfection reagent composition comprising:
 (a) 1,17-bis(2-octylcyclopropyl)heptadecan-9-yl-4-(dimethylamino)butanoate, or a pharmaceutically acceptable salt thereof;   (b) 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC);   (c) cholesterol; and   (d) one or more surfactants selected from the group consisting of polyoxyethylene (20) oleyl ether, polyoxyethylene (23) lauryl ether, and polyoxyethylene (40) stearate.   
     
     
         2 . The transfection reagent composition of  claim 1 , wherein the one or more surfactants is polyoxyethylene (20) oleyl ether. 
     
     
         3 . The transfection reagent composition of  claim 2 , wherein a molar ratio of 1,17-bis(2-octylcyclopropyl)heptadecan-9-yl-4-(dimethylamino)butanoate:DSPC:cholesterol:polyoxyethylene (20) oleyl ether is 50:10:38:2. 
     
     
         4 . The transfection reagent composition of  claim 1 , wherein the one or more surfactants is polyoxyethylene (23) lauryl ether. 
     
     
         5 . The transfection reagent composition of  claim 4 , wherein a molar ratio of 1,17-bis(2-octylcyclopropyl)heptadecan-9-yl-4-(dimethylamino)butanoate:DSPC:cholesterol:polyoxyethylene (23) lauryl ether is 50:10:38:2. 
     
     
         6 . The transfection reagent composition of  claim 1 , wherein the one or more surfactants is polyoxyethylene (40) stearate. 
     
     
         7 . The transfection reagent composition of  claim 6 , wherein a molar ratio of 1,17-bis(2-octylcyclopropyl)heptadecan-9-yl-4-(dimethylamino)butanoate:DSPC:cholesterol:polyoxyethylene (40) stearate is 50:10:37.5:2.5. 
     
     
         8 . A lipid nanoparticle comprising the transfection reagent composition of  claim 1  and an anionic macromolecule. 
     
     
         9 . The lipid nanoparticle of  claim 8 , wherein the lipid nanoparticle has a solid core. 
     
     
         10 . The lipid nanoparticle of  claim 8 , wherein the anionic macromolecule is selected from the group consisting of nucleic acids, anionic proteins, and anionic peptides. 
     
     
         11 . The lipid nanoparticle of  claim 8 , wherein the anionic macromolecule is a nucleic acid. 
     
     
         12 . The lipid nanoparticle of  claim 11 , wherein the nucleic acid is a DNA, an RNA, a locked nucleic acid, a nucleic acid analog, or a plasmid capable of expressing a DNA or an RNA. 
     
     
         13 . The lipid nanoparticle of  claim 11 , wherein the nucleic acid is an ssRNA, an siRNA, a microRNA, or an mRNA. 
     
     
         14 . The lipid nanoparticle of  claim 11 , wherein the nucleic acid is an antisense oligonucleotide. 
     
     
         15 . The lipid nanoparticle of  claim 11 , wherein the nucleic acid is a hybridization probe. 
     
     
         16 . The lipid nanoparticle of  claim 15 , wherein the hybridization probe is a molecular beacon.