Non-liposomal systems for nucleic acid delivery

Abstract

The present invention provides novel, stable lipid particles having a non-lamellar structure and comprising one or more active agents or therapeutic agents, methods of making such lipid particles, and methods of delivering and/or administering such lipid particles. More particularly, the present invention provides stable nucleic acid-lipid particles (SNALP) that have a non-lamellar structure and that comprise a nucleic acid (such as one or more interfering RNA), methods of making the SNALP, and methods of delivering and/or administering the SNALP.

1 - 32 . (canceled) 
     
     
         33 . A composition comprising:
 a plurality of nucleic acid-lipid particles, wherein each particle in the plurality of particles comprises:   (a) a nucleic acid;   (b) a cationic lipid comprising one or more diastereomers;   (c) a non-cationic lipid; and   (d) a conjugated lipid that inhibits aggregation of particles,   wherein at least about 95% of the particles in the plurality of particles are electron-dense.   
     
     
         34 . The composition of  claim 33 , wherein the nucleic acid is RNA. 
     
     
         35 . The composition of  claim 33 , wherein the nucleic acid is mRNA. 
     
     
         36 . The composition of  claim 33 , wherein the cationic lipid comprises a mixture of diastereomers. 
     
     
         37 . The composition of  claim 33 , wherein the cationic lipid consists of one or more diastereomers. 
     
     
         38 . The composition of  claim 33 , wherein the cationic lipid has a protonatable group with a pK a  of from about 4 to about 7. 
     
     
         39 . The composition of  claim 33 , wherein the non-cationic lipid is a mixture of (i) a phospholipid and cholesterol or (ii) a phospholipid and a cholesterol derivative. 
     
     
         40 . The composition of  claim 33 , wherein the conjugated lipid that inhibits aggregation of particles is a polyethyleneglycol (PEG)-lipid conjugate. 
     
     
         41 . The composition of  claim 40 , wherein the PEG-lipid conjugate is selected from the group consisting of a PEG-diacylglycerol (PEG-DAG) conjugate, a PEG dialkyloxypropyl (PEG-DAA) conjugate, a PEG-phospholipid conjugate, a PEG-ceramide (PEG-Cer) conjugate, and a mixture thereof. 
     
     
         42 . The composition of  claim 33 , wherein the electron-dense particles comprise an inverse hexagonal (Hn) or cubic phase structure. 
     
     
         43 . The composition of  claim 33 , wherein greater than 95% of the particles are electron-dense. 
     
     
         44 . The composition of  claim 33 , wherein at least 96% of the particles are electron-dense. 
     
     
         45 . The composition of  claim 33 , wherein at least 97% of the particles are electron-dense. 
     
     
         46 . The composition of  claim 33 , wherein at least 98% of the particles are electron-dense. 
     
     
         47 . The composition of  claim 33 , wherein at least 99% of the particles are electron-dense. 
     
     
         48 . The composition of  claim 33 , wherein the nucleic acid is fully encapsulated in the particles. 
     
     
         49 . The composition of  claim 33 , wherein the particles have a mean diameter of from about 60 nm to about 130 nm. 
     
     
         50 . The composition of  claim 33 , wherein the particles have a mean diameter of from about 70 nm to about 110 nm. 
     
     
         51 . The composition of  claim 33 , wherein the particles have a mean diameter of from about 90 nm to about 100 nm. 
     
     
         52 . The composition of  claim 33 , wherein the particles have a lipid:nucleic acid ratio of from about 2 to about 25. 
     
     
         53 . The composition of  claim 33 , wherein the particles have a lipid:nucleic acid ratio of from about 3 to about 20. 
     
     
         54 . A pharmaceutical composition comprising a composition of  claim 33  and a pharmaceutically acceptable carrier. 
     
     
         55 . A method for introducing a therapeutic agent into a cell, the method comprising:
 contacting the cell with a composition of  claim 33 .   
     
     
         56 . A method for the in vivo delivery of a therapeutic agent, the method comprising:
 administering to a mammal a composition of  claim 33 .