Processes of preparing mRNA-loaded lipid nanoparticles
AssigneeTranslate Bio, Inc.
InventorsShrirang Karve, Ashish Sarode, Frank DeRosa
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Abstract
The present invention provides an improved process for lipid nanoparticle formulation and mRNA encapsulation. In some embodiments, the present invention provides a process of encapsulating messenger RNA (mRNA) in lipid nanoparticles comprising a step of mixing a suspension of preformed lipid nanoparticles and mRNA.
Claims19
A process of encapsulating messenger RNA (mRNA) in preformed empty lipid nanoparticles (LNPs), wherein the lipid component of the 0.1:1NPs comprises or consists of a cationic lipid, a non-cationic lipid, a PEG-modified lipid, and optionally cholesterol and said process comprises the following steps:
- adding a first volume of a solution comprising the mRNA to a suspension comprising the preformed empty LNPs such that a mixture comprising LNPs encapsulating the MRNA is formed;
- adding one or more additional volumes of the solution comprising the mRNA to the mixture obtained in the preceding step until a desired molar ratio of cationic lipid to mRNA is achieved;
- and
- wherein the first volume and the one or more additional volumes of the solution comprising the mRNA are added continuously without intervening periods of heating or mixing.
A process of encapsulating messenger RNA (mRNA) in preformed empty lipid nanoparticles (LNPs), wherein the lipid component of the LNPs comprises or consists of a cationic lipid, a non-cationic lipid, a PEG-modified lipid, and optionally cholesterol and said process comprises the following steps:
- adding a first volume of a suspension comprising the preformed empty LNPs to a solution comprising the mRNA such that a mixture comprising LNPs encapsulating the mRNA is formed;
- adding one or more additional volumes of the suspension comprising the preformed empty LNPs to the mixture obtained in the preceding step until a desired molar ratio of cationic lipid to mRNA is achieved;
- and
- wherein the first volume and the one or more additional volumes of the suspension comprising the preformed empty LNPs are added continuously without intervening periods of heating or mixing.
A method of manufacturing a lipid nanoparticle (LNP) composition, the method comprising:
- (a) mixing a first set of preformed empty lipid nanoparticles (LNPs) comprising a first cationic lipid, a first non-cationic lipid, a first PEG-modified lipid, and optionally cholesterol with mRNA under conditions that allow encapsulation of the mRNA;
- (b) combining the mRNA-encapsulating LNPs formed in step (a) with a second set of preformed empty LNPs comprising a second cationic lipid, a second non-cationic lipid, a second PEG-modified lipid, and optionally cholesterol, resulting in the LNP composition.
Dependencies are inferred from the claim text.