IMPROVED NUCLEIC ACID LIPID PARTICLE FORMULATIONS

• US 20150306039A1
• Filed: 12/04/2013
• Published: 10/29/2015
• Est. Priority Date: 12/07/2012
• Status: Active Grant

• Current Assignee: Alnylam Pharmaceuticals Incorporated
• Original Assignee: Alnylam Pharmaceuticals Incorporated
• Inventors: AKINC, Akin, MAIER, Martin, KUMAR, Varun
• Granted Patent: US 9,687,448 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61K 9/1272 with substantial amounts of non-phosphatidyl, i.e. non-acylglycerophosphate, surfactants as bilayer-forming substances, e.g. cationic lipids with cholesterol as the only non-phosphatidyl surfactant A61K9/127; lipids as modifying agent A61K47/543
• CPC Class Codes: A61K 9/1617 Organic compounds, e.g. phospholipids, fats
• CPC Class Codes: A61K 9/1641 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poloxamers
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 15/88 using microencapsulation, e.g. using amphiphile liposome vesicle
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 2'-R Modification

Abstract

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

Claims

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 C₈ 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), dipalmitoylphosphatidylglycerol (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.

9. The lipid nanoparticles of claim 1, wherein the active pharmaceutical ingredient is a nucleic acid.

11. The lipid formulation of claim 1, wherein the ratio of lipid to nucleic acid is about 3 to about 15.

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.

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 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.

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 d₉₈ 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.

30. The formulation of claim 19, 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.

35. The formulation of claim 30, wherein the concentration of the isotonicity agent in the medium is at most about 300 mM.

40. The formulation of claim 19, wherein the lipid nanoparticles have a d₅₀ 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.

7-8. -8. (canceled)

10. The lipid nanoparticles of claim 10, wherein the active pharmaceutical ingredient is a siRNA.

12-13. -13. (canceled)

16. (canceled)

21. (canceled)

28-29. -29. (canceled)

32-34. -34. (canceled)

36-39. -39. (canceled)

42-44. -44. (canceled)