Nanostructured lipid carriers and stable emulsions and uses thereof
Provided herein are nanostructured lipid carrier compositions, and methods of making and using thereof. The compositions comprise a nanostructured lipid carrier (NLC), where the NLC comprises an oil core comprising a mixture of a liquid phase lipid and a solid phase lipid, a cationic lipid, a sorbitan ester, and a hydrophilic surfactant, and optionally a bioactive agent. The bioactive agent can be associated with the NLC. The compositions are capable of delivery of a biomolecule to a cell for the generation of an immune response, for example, for vaccine, therapeutic, or diagnostic uses. Compositions and methods related to making the compositions and using the compositions for stimulating an immune response are also provided.
1. A composition comprising nanostructured lipid carrier (NLC) particles for delivery of a bioactive agent to a cell, wherein the NLC particles comprise:
(a) an oil core comprising a mixture of a liquid phase lipid and a solid phase lipid that is trimyristin;
(b) a cationic lipid;
(c) a sorbitan ester;
(d) a hydrophilic surfactant; and
(e) the bioactive agent,
wherein relative amounts and specific components of the composition are selected such that when the bioactive agent is replicating viral RNA (rvRNA) encoding secreted alkaline phosphatase (SEAP), the highest peak expression of the bioactive agent occurs at a N:P molar ratio of 15 as measured by an in vitro SEAP expression assay using a SEAP-expressing replicon and measured by relative luminescence units (RLUs).
2. The composition of claim 1 , wherein the sorbitan ester is sorbitan monostearate.
3. The composition of claim 1 , wherein the bioactive agent is not encapsulated by the NLC particles and is associated with the surface of the NLC particles.
4. The composition of claim 1 , wherein the bioactive agent is a protein, encodes a protein, is a protein antigen, encodes a protein antigen, is RNA, is DNA, is mRNA, is oncolytic viral RNA, is non-coding RNA, or is an adjuvant.
5. The composition of claim 1 , wherein the average polydispersity index of the NLC particles is from 0.1 to about 0.5.
6. The composition of claim 1 , wherein the z-average diameter of the NLC particles is from about 40 nm to about 80 nm.
7. The composition of claim 1 , having an oil to surfactant molar ratio of about 0.5 to about 12.
8. The composition of claim 7 , having an oil to surfactant molar ratio of about 0.5.
9. The composition of claim 1 , having a hydrophilic surfactant to cationic component ratio of about 0.6.
10. A method of making the composition of claim 1 , comprising:
(a) mixing the solid phase lipid, the liquid phase lipid, the cationic lipid, and the sorbitan ester to form an oil phase mixture;
(b) mixing the hydrophilic surfactant and water to form an aqueous phase mixture; and
(c) mixing the oil phase mixture with the aqueous phase mixture to form the NLC particles.
11. A method of generating or enhancing an immune response comprising administering to a subject in need thereof a therapeutically effective amount of the composition of claim 1 , wherein the bioactive agent is an antigen or encodes an antigen.
12. The composition of claim 1 , further comprising a citrate buffer.
13. The composition of claim 1 , wherein the amount of hydrophilic surfactant is no greater than about 35% of the sum of the mass of the hydrophilic surfactant, sorbitan ester, and cationic lipid.
14. A composition comprising nanostructured lipid carrier (NLC) particles for delivery of a bioactive agent to a cell, wherein the NLC particles comprise:
(a) an oil core comprising a mixture of a liquid phase lipid that is squalene and a solid phase lipid that is trimyristin;
(b) a cationic lipid that is DOTAP;
(c) a sorbitan ester that is sorbitan monostearate;
(d) a hydrophilic surfactant that is polysorbate 80; and
(e) the bioactive agent,
wherein the NLC particles comprise about 3.75% w/v liquid phase lipid, about 0.25% w/v solid phase lipid, about w/v 3% cationic lipid, about 3.7% w/v sorbitan ester, and about 3.7% w/v hydrophilic surfactant.
15. The composition of claim 14 , wherein the bioactive agent is not encapsulated by the NLC particles and is associated with the surface of the NLC particles.
16. The composition of claim 14 , wherein the bioactive agent is a protein, encodes a protein, is a protein antigen, encodes a protein antigen, is RNA, is DNA, is mRNA, is oncolytic viral RNA, is non-coding RNA, or is an adjuvant.
17. The composition of claim 14 , wherein the average polydispersity index of the NLC particles is from 0.1 to about 0.5.
18. The composition of claim 14 , wherein the z-average diameter of the NLC particles is from about 40 nm to about 80 nm.
19. The composition of claim 14 , having an oil to surfactant molar ratio of about 0.5 to about 12.
20. The composition of claim 19 , having an oil to surfactant molar ratio of about 0.5.
21. The composition of claim 14 , having a hydrophilic surfactant to cationic lipid ratio of about 0.2 to about 1.5.
22. The composition of claim 21 , having a hydrophilic surfactant to cationic component ratio of about 0.6.
23. A method of making the composition of claim 14 , comprising:
(a) mixing the solid phase lipid, the liquid phase lipid, the cationic lipid, and the sorbitan ester to form an oil phase mixture;
(b) mixing the hydrophilic surfactant and water to form an aqueous phase mixture; and
(c) mixing the oil phase mixture with the aqueous phase mixture to form the NLC particles.
24. A method of generating or enhancing an immune response comprising administering to a subject in need thereof a therapeutically effective amount of the composition of claim 14 , wherein the bioactive agent is an antigen or encodes an antigen.
25. The composition of claim 14 , further comprising a citrate buffer.
26. A composition comprising nanostructured lipid carrier (NLC) particles for delivery of a bioactive agent to a cell, wherein the NLC particles comprise:
(a) an oil core comprising a mixture of a liquid phase lipid that is squalene and a solid phase lipid that is trimyristin;
(b) a cationic lipid that is DOTAP;
(c) a sorbitan ester that is sorbitan monostearate;
(d) a hydrophilic surfactant that is polysorbate 80; and
(e) the bioactive agent,
wherein a hydrophilic surfactant to cationic lipid molar ratio is about 0.2 to about 1.5.
27. The composition of claim 26 , wherein the bioactive agent is not encapsulated by the NLC particles and is associated with the surface of the NLC particles.
28. The composition of claim 26 , wherein the bioactive agent is a protein, encodes a protein, is a protein antigen, encodes a protein antigen, is RNA, is DNA, is mRNA, is oncolytic viral RNA, is non-coding RNA, or is an adjuvant.
29. The composition of claim 26 , wherein the average polydispersity index of the NLC particles is from 0.1 to about 0.5.
30. The composition of claim 26 , wherein the z-average diameter of the NLC particles is from about 40 nm to about 80 nm.
31. The composition of claim 26 , having an oil to surfactant molar ratio of about 0.5 to about 12.
32. The composition of claim 31 , having an oil to surfactant molar ratio of about 0.5.
33. The composition of claim 26 , having a hydrophilic surfactant to cationic component ratio of about 0.6.
34. A method of making the composition of claim 26 , comprising:
(a) mixing the solid phase lipid, the liquid phase lipid, the cationic lipid, and the sorbitan ester to form an oil phase mixture;
(b) mixing the hydrophilic surfactant and water to form an aqueous phase mixture; and
(c) mixing the oil phase mixture with the aqueous phase mixture to form the NLC particles.
35. A method of generating or enhancing an immune response comprising administering to a subject in need thereof a therapeutically effective amount of the composition of claim 26 , wherein the bioactive agent is an antigen or encodes an antigen.
36. The composition of claim 26 , further comprising a citrate buffer.
37. The composition of claim 26 , wherein the amount of hydrophilic surfactant is no greater than about 35% of the sum of the mass of the hydrophilic surfactant, sorbitan ester, and cationic lipid.
38. A composition comprising nanostructured lipid carrier (NLC) particles for delivery of a bioactive agent to a cell, wherein the NLC particles comprise:
(a) an oil core comprising a mixture of a liquid phase lipid and a solid phase lipid that is trimyristin;
(b) a cationic lipid;
(c) a sorbitan ester;
(d) a hydrophilic surfactant; and
(e) the bioactive agent,
wherein the composition is formulated such that when administered to mice in which the bioactive agent is rvRNA encoding ZIKV prM/E from a Venezuelan equine encephalitis virus (VEEV) plasmid, the composition induces mean PRNT 80 titers of between about 1:110 and about 1:2560 when loaded with about 30 ng to about 100 ng of the rvRNA at a N:P molar ratio of about 15.
39. The composition of claim 38 , wherein the bioactive agent is not encapsulated by the NLC particles and is associated with the surface of the NLC particles.
40. The composition of claim 38 , wherein the bioactive agent is a protein, encodes a protein, is a protein antigen, encodes a protein antigen, is RNA, is DNA, is mRNA, is oncolytic viral RNA, is non-coding RNA, or is an adjuvant.
41. The composition of claim 38 , wherein the average polydispersity index of the NLC particles is from 0.1 to about 0.5.
42. The composition of claim 38 , wherein the z-average diameter of the NLC particles is from about 40 nm to about 80 nm.
43. The composition of claim 38 , having an oil to surfactant molar ratio of about 0.5 to about 12.
44. The composition of claim 43 , having an oil to surfactant molar ratio of about 0.5.
45. The composition of claim 38 , having a hydrophilic surfactant to cationic component ratio of about 0.6.
46. A method of making the composition of claim 38 , comprising:
(a) mixing the solid phase lipid, the liquid phase lipid, the cationic lipid, and the sorbitan ester to form an oil phase mixture;
(b) mixing the hydrophilic surfactant and water to form an aqueous phase mixture; and
(c) mixing the oil phase mixture with the aqueous phase mixture to form the NLC particles.
47. A method of generating or enhancing an immune response comprising administering to a subject in need thereof a therapeutically effective amount of the composition of claim 38 , wherein the bioactive agent is an antigen or encodes an antigen.
48. The composition of claim 38 , further comprising a citrate buffer.
49. The composition of claim 38 , wherein the amount of hydrophilic surfactant is no greater than about 35% of the sum of the mass of the hydrophilic surfactant, sorbitan ester, and cationic lipid.