Compositions for the in vivo delivery of RNAi agents
View Patent ↗This application describes emulsion formulations containing neutral phospholipids for delivering RNAi, anti-miRNA, or aptamer agents in vivo. The application also relates to methods of making the formulations, and uses of the formulations as delivery agents.
1. A composition for the in vivo delivery of an RNAi agent, comprising an emulsion of a lipid component, an aqueous component, and a non-ionic surfactant, wherein
the lipid component comprises 20-40% by weight of a neutral phospholipid and 60-80% by weight of an oil or wax, and wherein the percentage by weight of neutral phospholipid and the percentage by weight of oil or wax totals 100% of the lipid component;
the aqueous component comprises an RNAi agent in an aqueous medium, wherein the RNAi agent is less than 200 nucleotides in length; and
the surfactant comprises 0.1-50% of the total emulsion by weight; wherein the composition does not comprise a cationic lipid.
2. The composition of claim 1 , wherein the neutral phospholipid is 1,2-dioleoyl-sn-glycero-3-phosphocholine.
3. The composition of claim 1 or 2 , wherein the oil is squalene.
4. The composition of claim 3 , wherein the surfactant is polysorbate 20.
5. The composition of claim 3 , wherein the RNAi agent is a miRNA.
6. The composition of claim 3 , wherein the RNAi agent is an siRNA.
7. The composition of claim 1 , further comprising an antioxidant.
8. The composition of claim 1 , wherein the surfactant comprises 40-50% of the total emulsion by weight.
9. A method of making an emulsion comprising the steps of:
a. mixing a neutral phospholipid, an oil, and a non-ionic surfactant in an organic solvent;
b. evaporating the organic solvent to form a dried lipid component; and
c. mixing the lipid component with an RNAi agent in an aqueous medium;
wherein
the lipid component comprises 20-40% by weight of a neutral phospholipid and 60-80% by weight of an oil or wax and wherein the percentage by weight of neutral phospholipid and the percentage by weight of oil or wax totals 100% of the lipid component;
the RNAi component comprises an RNAi agent in an aqueous medium, wherein the RNAi agent is less than 200 nucleotides in length; and
the surfactant comprises 0.1-50% of the total emulsion by weight,
wherein the emulsion does not comprise a cationic lipid.
10. The method of claim 9 , wherein the neutral phospholipid is 1,2-dioleoyl-sn-glycero-3-phosphocholine.
11. The method of claim 9 or 10 , wherein the oil is squalene.
12. The method of claim 11 , wherein the surfactant is polysorbate 20.
13. The method of claim 11 , wherein the RNAi agent is a miRNA.
14. The method of claim 11 , wherein the RNAi agent is an siRNA.
15. The method of claim 9 , wherein lipid component comprises 20-40% phospholipid and 60-80% oil or wax; and the surfactant comprises 40-50% of the total emulsion by weight.
16. The method of claim 9 , wherein the evaporating is performed by lyophilization.
17. The method of claim 9 , wherein the mixing in step c is performed by sonication.
18. The method of claim 9 , further comprising extruding the mixture formed in step c.
19. The method of claim 9 , further comprising adding an antioxidant to the mixture of step a.
20. A method of delivering an RNAi agent to an animal, comprising administering the composition of claim 1 to the animal.
21. The method of claim 20 , wherein the composition is administered by injection.
22. The method of claim 21 , wherein the composition is administered systemically.
23. The method of claim 21 , wherein the RNAi agent is a miRNA.
24. A composition comprising an emulsion of a lipid component, an aqueous component, and a non-ionic surfactant, wherein
the lipid component comprises 20-40% by weight of a neutral phospholipid and 60-80% by weight of an oil or wax, and wherein the percentage by weight of neutral phospholipid and the percentage by weight of oil or wax totals 100% of the lipid component;
the aqueous component comprises an anti-miRNA agent in an aqueous medium; and
the surfactant comprises 0.1-50% of the total emulsion by weight; wherein the composition does not comprise a cationic lipid.
25. A composition comprising an emulsion of a lipid component, an aqueous component, and a non-ionic surfactant, wherein
the lipid component comprises 20-40% by weight of a neutral phospholipid and 60-80% by weight of an oil or wax, and wherein the percentage by weight of neutral phospholipid and the percentage by weight of oil or wax totals 100% of the lipid component;
the aqueous component comprises an aptamer in an aqueous medium; and
the surfactant comprises 0.1-50% of the total emulsion by weight; wherein the composition does not comprise a cationic lipid.
26. A composition for the in vivo delivery of an RNAi agent, comprising an emulsion of a lipid component, an aqueous component, and polysorbate 20, wherein
the lipid component comprises 20-40% by weight of 1,2-dioleoyl-sn-glycero-3-phosphocholine and 60-80% by weight of squalene, and wherein the percentage by weight of 1,2-dioleoyl-sn-glycero-3-phosphocholine and the percentage by weight of squalene totals 100% of the lipid component;
the aqueous component comprises an RNAi agent in an aqueous medium, wherein the RNAi agent is less than 200 nucleotides in length; and
the polysorbate 20 comprises 0.1-50% of the total emulsion by weight; wherein the composition does not comprise a cationic lipid.
27. The composition of claim 26 , wherein the polysorbate 20 comprises 40-50% of the total emulsion by weight.
28. A composition for the in vivo delivery of an RNAi agent, comprising an emulsion of a lipid component, an aqueous component, and a non-ionic surfactant, wherein
the lipid component comprises a neutral phospholipid and an oil or wax, wherein the ratio of neutral phospholipid to oil or wax is from 1:2 to 1:6, and wherein the lipid component comprises only neutral phospholipid and oil or wax;
the aqueous component comprises an RNAi agent in an aqueous medium, wherein the RNAi agent is less than 200 nucleotides in length; and
the surfactant comprises 0.1-50% of the total emulsion by weight; wherein the composition does not comprise a cationic lipid.
29. The composition of claim 28 , wherein the neutral phospholipid is 1,2-dioleoyl-sn-glycero-3-phosphocholine, the oil or wax is squalene, and the surfactant is polysorbate 20.