Process and formulation of lipid nanoparticles
AssigneeTranslate Bio, Inc.
InventorsFrank DEROSA, Hardip R. GOPANI, Shrirang KARVE
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Abstract
The present invention provides an improved process for preparing a stable lyophilized composition comprising lipid nanoparticles encapsulating mRNA, which results in high mRNA integrity suitable for both short- and long-term storage for therapeutic use. In some embodiments, the present invention provides a method of preparing a stable lyophilized composition comprising the steps of adding ascorbic acid to an aqueous solution comprising one or more lipid nanoparticles, freezing the aqueous solution to obtain a frozen solution, drying the frozen solution to obtain a lyophilized composition, and increasing and holding the temperature of the lyophilized composition to a temperature of between 15° C. to 30° C.
Claims39
A method of preparing a stable lyophilized composition comprising one or more lipid nanoparticles encapsulating mRNA, the method comprising the steps of:
- (a) adding between 5 mM and 200 mM ascorbic acid to an aqueous solution comprising one or more lipid nanoparticles encapsulating mRNA having an initial mRNA integrity,
- (b) freezing the aqueous solution to obtain a frozen solution,
- (c) drying the frozen solution for a period of greater than 10 hours to obtain a lyophilized composition,
- wherein upon reconstitution of the lyophilized composition following storage of the lyophilized composition for four weeks at a temperature of 15 to 25° C., the mRNA integrity following reconstitution is at least 50% the initial mRNA integrity.
The method of any one of the preceding claims, wherein the mRNA integrity following reconstitution is at least 60%, at least 70%, at least 80%, or at least 85% the initial mRNA integrity.
The method of any one of the preceding claims, wherein the method includes, following step (c),
- (d) increasing and holding the temperature of the lyophilized composition to a temperature of between 15° C. to 30° C.
The method of any one of the preceding claims, wherein the aqueous solution comprises about 10 mM to 50 mM ascorbic acid.
The method of any one of the preceding claims, wherein the drying step is performed for longer than 20, 50, 100, 120, or 140 hours.
The method of any one of the preceding claims, wherein the lyophilized composition is held at a temperature of between 15° C. to 30° C. for longer than 1 hour, 5 hours, or 10 hours in step (d).
The method of any one of the preceding claims, wherein the initial mRNA integrity is higher than 50% and the mRNA integrity following reconstitution is higher than 40%.
The method of any one of the preceding claims, wherein the initial mRNA integrity is higher than 40% and the mRNA integrity following reconstitution remains higher than 40%.
The method of any one of the preceding claims, wherein the initial mRNA integrity is higher than 50% and the mRNA integrity following reconstitution remains higher than 50%.
The method of any one of the preceding claims, wherein the mRNA integrity is determined by capillary electrophoresis.
The method of any one of the preceding claims, wherein the drying step is performed at a temperature of between −20 and −30° C.
The method of any one of the preceding claims, wherein the drying step is performed at a pressure of about 50 mTorr.
The method of any one of the preceding claims wherein the temperature of the lyophilized composition is increased and held at a temperature of about 20° C. in step (d).
The method of any one of the preceding claims, wherein the aqueous solution comprises trehalose.
The method of any one of the preceding claims, wherein the aqueous solution has a pH below 7.0.
A stable lyophilized composition comprising one or more lipid nanoparticles encapsulating mRNA prepared by the method of any one of the preceding claims.
A stable lyophilized composition comprising one or more lipid nanoparticles encapsulating mRNA and between 5 mM and 200 mM ascorbic acid, wherein upon reconstitution of the lyophilized composition following storage of the lyophilized composition for four weeks at a temperature of 15 to 25° C., the integrity of the mRNA following reconstitution is at least 50% the initial mRNA integrity.
Dependencies are inferred from the claim text.