Method of lyophilizing lipid nanoparticles
Methods of preparing lyophilized lipid nanoparticle-nucleic acid compositions are provided. The methods comprise preparing a suspension of lipid nanoparticles with a monosaccharide and one or more excipients selected from thiosulfate, potassium sorbate, sodium benzoate, and iodixanol. Lyophilized lipid nanoparticle-nucleic acid compositions and methods of reconstituting and administering the same are further provided.
1. A pharmaceutical composition comprising a solid lyophilized composition, wherein the solid lyophilized composition comprises:
i. lipid nanoparticles encapsulating from about 0.001 to about 1.0% w/w of a ribonucleic acid (RNA) by the weight of the lyophilized composition,
ii. a lyoprotectant composition comprising from about 0.01% w/w to about 1.0% w/w of a poloxamer by the weight of the lyophilized composition and from about 0.5% w/w to about 5.0% w/w of potassium sorbate by the weight of the lyophilized composition, and
iii. from about 85% w/w to about 96% w/w of a sugar by the weight of the lyophilized composition.
2. The pharmaceutical composition of claim 1 , wherein the poloxamer is poloxamer 188.
3. The pharmaceutical composition of claim 1 , wherein the solid lyophilized composition comprises from about 0.5 to about 5.0% w/w of lipids by the weight of the solid lyophilized composition.
4. The pharmaceutical composition of claim 1 , wherein the solid lyophilized composition further comprises from about 0.5 to about 2.5% w/w of tris(hydroxymethyl)aminomethane by the weight of the solid lyophilized composition.
5. The pharmaceutical composition of claim 1 , wherein the solid lyophilized composition further comprises from about 0.75 to about 2.75% w/w of NaCl by the weight of the solid lyophilized composition.
6. The pharmaceutical composition of claim 1 , wherein the sugar is sucrose.
7. A method of preserving the solid lyophilized composition of claim 1 comprising storing the lyophilized product at a temperature of about 2° C. to about 8° C.
8. A method of preserving the solid lyophilized composition of claim 1 comprising storing the lyophilized product at a temperature of about −20° C.
9. A method of reconstituting the solid lyophilized composition of claim 1 comprising adding a liquid medium to the lyophilized composition.
10. The method of claim 9 , wherein the liquid medium is an aqueous medium.
11. The method of claim 9 , wherein the liquid medium further comprises a buffer having a pH of about 7.0 to about 8.5.
12. A method of treating a disease or disorder in a subject comprising administering to the subject the solid lyophilized composition of claim 1 reconstituted in a liquid medium.
13. The method of claim 12 , wherein the reconstituted lyophilized composition is administered intravenously, mucosally, or subcutaneously.
14. The method of claim 12 , wherein the reconstituted lyophilized composition is administered intramuscularly.
15. The method of claim 12 , wherein the reconstituted lyophilized composition is administered via inhalation.
16. A pharmaceutical composition comprising a solid lyophilized composition, wherein the solid lyophilized composition comprises:
i. lipid nanoparticles encapsulating from about 0.001 to about 1.0% w/w of an RNA by the weight of the lyophilized composition,
ii. from about 0.01% w/w to about 1.0% w/w of a poloxamer by the weight of the lyophilized composition and from about 0.5% w/w to about 5.0% w/w of potassium sorbate by the weight of the lyophilized composition,
iiii. from about 0.75% w/w to about 2.75% w/w of a salt by the weight of the lyophilized composition, and
iv. from about 85% w/w to about 96% w/w of a sugar by the weight of the lyophilized composition,
wherein upon reconstitution of the solid lyophilized composition with a liquid medium the lipid nanoparticles have a change in mean diameter less than about 15 nm as compared to a mean diameter prior to lyophilization.