Nucleic acid lipid particle formulations

Abstract

The present invention relates to lipid nanoparticles containing a biodegradable cationic lipid which provide improved delivery of active pharmaceutical ingredients, such as siRNA.

1 . Lipid nanoparticles comprising:
 (a) a biodegradable cationic lipid;   (b) polyethylene glycol-dipalmitoylglycerol (PEG-DPG);   (c) a non-cationic lipid (such as a neutral lipid);   (d) optionally, a sterol; and   (e) an active pharmaceutical ingredient.   
     
     
         2 . The lipid nanoparticles of  claim 1 , wherein the cationic lipid has a pKa ranging from about 5 to about 7. 
     
     
         3 . The lipid nanonparticles of  claim 1 , wherein the cationic lipid has
 (i) a central atom (e.g., a carbon, nitrogen, or phosphorous central atom),   (ii) an nitrogen containing head group bound to the central atom, and   (iii) two hydrophobic tails directly bound to the central atom, each hydrophobic tail comprising a C 8  or greater aliphatic group attached to the central atom, where the aliphatic group (a) is interrupted by a biodegradable group such that there is a chain of at least four carbon atoms between the biodegradable group and the central atom, or (b) includes a biodegradable group at the terminal end of the hydrophobic tail.   
     
     
         4 . The lipid nanoparticles of  claim 1 , wherein the sterol is cholesterol. 
     
     
         5 . The lipid nanoparticles of  claim 1 , wherein the non-cationic lipid comprises a phospholipid. 
     
     
         6 . The lipid nanoparticles of  claim 1 , wherein the non-cationic lipid is distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dip almitoylpho sphatidylglycerol (DPPG), dioleoyl-phosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoylphosphatidylethanolamine (POPE), dioleoyl- phosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), dimyristoyl phosphatidylcholine (DMPC), distearoyl-phosphatidyl-ethanolamine (DSPE), SM, 16-O-monomethyl PE, 16-O-dimethyl PE, 18-1-trans PE, 1-stearoyl-2-oleoyl-phosphatidyethanolamine (SOPE), cholesterol, or a mixture thereof. 
     
     
         7 . The lipid nanoparticles of  claim 7 , wherein the non-cationic lipid is distearoylphosphatidylcholine (DSPC). 
     
     
         8 . The lipid nanoparticles of  claim 1 , wherein the sterol is cholesterol. 
     
     
         9 . The lipid nanoparticles of  claim 1 , wherein the active pharmaceutical ingredient is a nucleic acid. 
     
     
         10 . The lipid nanoparticles of  claim 10 , wherein the active pharmaceutical ingredient is a siRNA. 
     
     
         11 . The lipid formulation of  claim 1 , wherein the ratio of lipid to nucleic acid is about 3 to about 15. 
     
     
         12 . The lipid formulation of  claim 11 , wherein the ratio of lipid to nucleic acid about 5 to about 13. 
     
     
         13 . The lipid nanoparticles of  claim 1 , wherein the nucleic acid is fully encapsulated in the lipid nanoparticle. 
     
     
         14 . The lipid nanoparticles of  claim 1 , wherein the PEG-DPG is present in less than about 3 mol% in the lipid nanoparticles, based upon the total moles of lipid in the lipid nanoparticles. 
     
     
         15 . The lipid nanoparticles of  claim 1 , wherein the lipid nanoparticles comprise from about 45 to about 65% of biodegradable cationic lipid, from about 5 to about 10% of a non-cationic lipid, from about 25 to about 40% of a sterol, and from about 0.5 to about 5% of PEG-DPG, based upon 100% total moles of lipid in the lipid nanoparticles. 
     
     
         16 . The lipid nanoparticles of  claim 1 , wherein the molar ratio of biodegradable cationic lipid, non-cationic lipid, sterol, and PEG-DPG is about 50:10:38.5:1.5. 
     
     
         17 . The lipid nanoparticles of  claim 1 , wherein the lipid nanoparticles are prepared by an in-line mixing method. 
     
     
         18 . The formulation of  claim 1 , wherein the lipid nanoparticles exhibit a single mode size distribution. 
     
     
         19 . A pharmaceutical formulation for parenteral administration comprising
 (a) lipid nanoparticles of  claim 1  any of the preceding claims, in   (b) a medium, wherein the formulation has one or more of the following characteristics:
 (i) the medium is substantially free of anions, 
 (ii) the medium is non-ionic or substantially non-ionic, and 
 (iii) the formulation has a pH less than the pKa of the cationic lipid. 
   
     
     
         20 . The formulation of  claim 19 , wherein the medium comprises a non-ionic diluent. 
     
     
         21 . The formulation of  claim 20 , wherein the non-ionic diluent increases the stability of the lipid nanoparticle thereby preventing aggregation. 
     
     
         22 . The formulation of  claim 19 , wherein the medium comprises water. 
     
     
         23 . The formulation of  claim 22 , wherein the water in the medium has been purified by reverse osmosis. 
     
     
         24 . The formulation of  claim 22 , wherein the water in the medium is deionized. 
     
     
         25 . The formulation of  claim 19 , wherein the medium contains less than 100 ppm of mineral acids. 
     
     
         26 . The formulation of  claim 19 , wherein the lipid nanoparticles have a d 98  of less than about 150 nm. 
     
     
         27 . The formulation of  claim 19 , wherein the formulation further comprises an acid, and the ratio of (a) the anion concentration from the acid to (b) the acid is less than about 0.5. 
     
     
         28 . The formulation of  claim 27 , wherein the ratio is less than about 0.3. 
     
     
         29 . The formulation of  claim 27 , wherein the ratio is less than about 0.2. 
     
     
         30 . The formulation of  claim 1 , wherein the formulation further comprises a nonionic or substantially non-ionic isotonicity agent. 
     
     
         31 . The formulation of  claim 30 , wherein the isotonicity agent is a polyol, sugar, amino acid, or albumin. 
     
     
         32 . The formulation of  claim 31 , wherein said the isotonicity agent is selected from glucose, dextrose, mannitol, sorbitol, trehalose, amino acid, albumin, and combinations thereof. 
     
     
         33 . The formulation of  claim 32 , wherein the isotonicity agent is glucose. 
     
     
         34 . The formulation of  claim 30 , wherein the amount of the isotonicity agent is sufficient for the formulation to obtain an isotonic level. 
     
     
         35 . The formulation of  claim 30 , wherein the concentration of the isotonicity agent in the medium is at most about 300 mM. 
     
     
         36 . (canceled) 
     
     
         37 . The formulation of  claim 35 , wherein the lipid nanoparticles have a d 99  of less than about 500 nm. 
     
     
         38 . The formulation of  claim 37 , wherein the lipid nanoparticles have a d 99  of less than about 250 nm. 
     
     
         39 . The formulation of  claim 38 , wherein the lipid nanoparticles have a d 99  of less than about 100 nm. 
     
     
         40 . The formulation of  claim 19 , wherein the lipid nanoparticles have a d 50  of less than about 50 nm. 
     
     
         41 . The formulation of  claim 19 , wherein the lipid nanoparticles has a mean diameter of less than about 100 nm after about 1 month at 4° C. 
     
     
         42 . The formulation of  claim 19 , wherein the lipid nanoparticles have an encapsulation efficiency of greater than about 90% after about 1 month at 4° C. 
     
     
         43 . The formulation of  claim 19 , wherein the formulation is a solution. 
     
     
         44 . The formulation of  claim 19 , wherein the formulation is a suspension.