Nucleic acid-containing lipid particles and related methods

Abstract

Lipid particles containing a nucleic acid, devices and methods for making the lipid particles, and methods for using the lipid particles.

1 . A method for introducing a polynucleic acid into a cell, comprising contacting a cell with a lipid particle, comprising an electron dense core comprising a polynucleic acid and an ionizable lipid, the core coated with one or more polar lipids comprising a PEG-lipid. 
     
     
         2 . The method of  claim 1 , wherein the ionizable lipid is an amino lipid. 
     
     
         3 . The method of  claim 1 , wherein the ionizable lipid is a dilinoleyl amino lipid. 
     
     
         4 . The method of  claim 1 , wherein the ionizable lipid is selected from the group consisting of DODAC, DOTMA, DDAB, DOTAP, DOTAP.Cl, DC-Chol, DOSPA, DOGS, DOPE, DODAP, DODMA, and DMRIE. 
     
     
         5 . The method of  claim 1 , wherein the ionizable lipid has the formula: 
       
         
           
           
               
               
           
         
         wherein R 1  and R 2  are either the same or different and independently optionally substituted C 10 -C 24  alkyl, optionally substituted C 10 -C 24  alkenyl, optionally substituted C 10 -C 24  alkynyl, or optionally substituted C 10 -C 24  acyl; 
         R 3  and R 4  are either the same or different and independently optionally substituted C 1 -C 6  alkyl, optionally substituted C 2 -C 6  alkenyl, or optionally substituted C 2 -C 6  alkynyl or R 3  and R 4  may join to form an optionally substituted heterocyclic ring of 4 to 6 carbon atoms and 1 or 2 heteroatoms chosen from nitrogen and oxygen; 
         R 5  is either absent or present and when present is hydrogen or C 1 -C 6  alkyl; 
         m, n, and p are either the same or different and independently either 0 or 1 with the proviso that m, n, and p are not simultaneously 0; 
         q is 0, 1 , 2, 3, or 4; and 
         Y and Z are either the same or different and independently O, S, or NH. 
       
     
     
         6 . The method of  claim 1 , wherein the PEG-lipid is selected from the group consisting of PEG-modified phosphatidylethanolamines, PEG-modified phosphatidic acids, PEG-modified ceramides, PEG-modified dialkylamines, PEG-modified diacylglycerols, and PEG-modified dialkylglycerols. 
     
     
         7 . The method of  claim 1 , wherein the PEG-lipid is selected from the group consisting of PEG-c-DMA, PEG-c-DOMG, and PEG-s-DMG. 
     
     
         8 . The method of  claim 1 , comprising from about 1 to about 5 mole percent PEG-lipid. 
     
     
         9 . The method of  claim 1 , comprising about 1 mole percent PEG-lipid. 
     
     
         10 . The method of  claim 1 , wherein the polar lipids are selected from the group consisting of neutral lipids and sterols. 
     
     
         11 . The method of  claim 1 , wherein the polar lipids comprise a neutral lipid selected from the group consisting of diacylphosphatidylcholines, diacylphosphatidylethanolamines, ceramides, sphingomyelins, dihydrosphingomyelins, cephalins, and cerebrosides. 
     
     
         12 . The method of  claim 1 , wherein the polar lipids comprise a neutral lipid selected from DSPC and DOPC. 
     
     
         13 . The method  claim 1 , wherein the polar lipids comprise a sterol. 
     
     
         14 . The method of  claim 1 , wherein the polynucleic 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. 
     
     
         15 . The method of  claim 1 , wherein the polynucleic acid is ssDNA or dsDNA. 
     
     
         16 . The method of  claim 1 , wherein the polynucleic acid is siRNA or microRNA. 
     
     
         17 . The method particle of  claim 1 , wherein the polynucleic acid is an oligonucleotide. 
     
     
         18 . The method of  claim 1 , wherein the polynucleic acid is an antisense oligonucleotide. 
     
     
         19 . The method of  claim 1 , wherein the particle has a diameter from about 30 nm to about 200 nm. 
     
     
         20 . The method of  claim 1 , wherein the particle has a diameter of about 80 nm.