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, wherein the liquid phase lipid is squalene and the solid phase lipid is trimyristin;
(b) a cationic component, when the cationic component is DOTAP;
(c) a hydrophobic surfactant, wherein the hydrophobic surfactant is sorbitan monostearate;
(d) a hydrophilic surfactant wherein the hydrophilic surfactant is polysorbate 80; and
(e) the bioactive agent, wherein the bioactive agent is RNA, mRNA, oncolytic viral RNA, non-coding RNA, or DNA,
wherein a hydrophilic surfactant to cationic component molar ratio is about 0.2 to about 1.5.
2. 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.
3. The composition of claim 1 , wherein the composition delivers the bioactive agent to the cell.
4. The composition of claim 1 , wherein the hydrophobic surfactant is present in an amount sufficient to increase the ability of the composition to deliver the bioactive agent to the cell as compared to a comparable composition without the hydrophobic surfactant.
5. The composition of claim 1 , wherein the cell is in a subject, wherein the bioactive agent is an antigen or encodes an antigen, and wherein the composition elicits an immune response in the subject against the antigen.
6. The composition of claim 5 , wherein the composition induces neutralizing antibody titers in the subject at a higher level than the neutralizing antibody titers induced in the subject by a comparable composition lacking the hydrophobic surfactant.
7. The composition of claim 1 , wherein the bioactive agent is RNA, and wherein said RNA encodes an antigen or an antibody.
8. The composition of claim 1 , wherein the average polydispersity index of the NLC particles is from 0.1 to about 0.5.
9. The composition of claim 1 , wherein the z-average diameter of the NLC particles is from about 40 nm to about 80 nm.
10. The composition of claim 1 , having an oil to surfactant molar ratio of about 0.05 to about 12.
11. The composition of claim 1 , wherein the hydrophilic surfactant to cationic component molar ratio is about 0.5 to about 1.
12. The composition of claim 1 , comprising from about 0.2% to about 40% w/v liquid phase lipid, from about 0.1% to about 10% w/v solid phase lipid, from about 0.2% to about 10% w/v cationic component as a cationic lipid, from about 0.25% to about 15% w/v hydrophobic surfactant, and from about 0.2% to about 15% w/v hydrophilic surfactant.
13. The composition of claim 1 , wherein the bioactive agent is RNA, and wherein the RNA encodes one or more TB antigens.
14. The composition of claim 1 , further comprising an adjuvant.
15. The composition of claim 14 , wherein the adjuvant is selected from a TLR agonist, a Rig-I agonist, a saponin, a carbohydrate, a carbohydrate polymer, a conjugated carbohydrate, a whole viral particle, a virus-like particle, viral fragments, and cellular fragments.
16. The composition of claim 11 , wherein the hydrophilic surfactant to cationic component molar ratio is about 0.6.
17. The composition of claim 10 , wherein the oil to surfactant molar ratio is about 0.5 to about 8.
18. The composition of claim 17 , wherein the oil to surfactant molar ratio is about 0.5.
19. A method of making the composition of claim 1 , comprising:
(a) mixing the solid phase lipid, the liquid phase lipid, the cationic component, and the hydrophobic surfactant to form an oil phase mixture;
(b) mixing the hydrophilic surfactant and water to form an aqueous phase mixture;
(c) mixing the oil phase mixture with the aqueous phase mixture to form the NLC particles; and
(d) combining the bioactive agent with the NLC particles such that the bioactive agent associates with the surface of the NLC particles by non-covalent interactions or by reversible covalent interactions.
20. 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 .
21. A method of delivering a bioactive agent to a cell, comprising contacting the cell with the composition of claim 1 .
22. A method of generating an immune response in a subject, comprising
(a) administering to a subject in need thereof a therapeutically effective amount of an oncolytic virus encoding a protein antigen, and
(b) administering to the subject a therapeutically effective amount of the NLC particles of claim 1
wherein the bioactive agent is a nucleic acid molecule encoding a protein antigen.
23. The method of claim 22 , wherein the administration of (a) and the administration of (b) occur at least 1 week, at least 2 weeks, at least 3 weeks, at least 1 month, at least 6 weeks, at least two months, at least three months, at least 6 months, or at least 1 year apart.
24. The method of claim 22 , wherein the bioactive agent is RNA, and wherein the RNA encodes one or more TB antigens.