Methods for purification of ionizable lipids
AssigneeModernaTX, Inc.
InventorsGabor Butora, Alison Volkert, Dipendra Gyawali, Atsushi Endo, Zhong Yang, Jin Lim
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Abstract
Provided herein are methods for purifying an ionizable amino lipid (IAL) composition comprising impurities that may react with polynucleotides, such as mRNA. Methods for purifying IAL compositions comprise one or more scavenging-removal steps to selectively remove reactive impurities.
Claims77
A method of purifying an ionizable amino lipid (IAL) present in a first composition comprising the IAL, a first reactive impurity, and a second reactive impurity in a first nonpolar solvent, the method comprising:
- (a) performing a first scavenging-removal step, comprising:
- contacting the first composition with a first scavenger, wherein the first scavenger reacts with the first reactive impurity to form a first scavenger-impurity product;
- and
- separating the first scavenger-impurity product from the first composition to obtain a second composition comprising the IAL and the second reactive impurity;
- (b) performing an acid extraction step, comprising contacting the second composition with an acid, wherein the acid reacts with the IAL to form an IAL salt that is soluble in a polar solvent, and transferring the IAL salt and the second reactive impurity into the polar solvent to obtain an IAL salt solution;
- (c) performing a second scavenging-removal step, comprising:
- contacting the IAL salt solution with a second scavenger, wherein the second scavenger reacts with the second reactive impurity to form a second scavenger-impurity product;
- converting the IAL salt into the IAL with a neutralizing agent, wherein the IAL is soluble in a second nonpolar solvent;
- and
- separating the IAL from the second scavenger-impurity product, or a derivative thereof formed by the neutralizing agent, by transferring the IAL into the second nonpolar solvent to obtain a purified IAL composition comprising purified IAL.
A method of purifying an ionizable amino lipid (IAL) present in a first composition comprising the IAL, a reactive halide impurity, and a reactive aldehyde impurity in a first nonpolar solvent, the method comprising:
- (a) performing a halide scavenging-removal step, comprising:
- contacting the first composition with a halide scavenger that reacts with the reactive halide impurity to form a scavenger-halide product;
- and
- separating the scavenger-halide product from the first composition to obtain a second composition comprising the IAL and reactive aldehyde impurity;
- (b) performing an acid extraction step, comprising contacting the second composition with an acid that reacts with the IAL to form an IAL salt that is soluble in a polar solvent, and transferring the IAL salt and reactive aldehyde impurity into the polar solvent to obtain an IAL salt solution;
- (c) performing an aldehyde scavenging-removal step, comprising:
- contacting the IAL salt solution with an aldehyde scavenger that reacts with the reactive aldehyde impurity to form a scavenger-aldehyde product;
- converting the IAL salt into the IAL with a neutralizing agent, wherein the IAL is soluble in a second nonpolar solvent;
- and
- separating the IAL from the scavenger-aldehyde product, or a derivative thereof formed by the neutralizing agent, by transferring the IAL into the second nonpolar solvent to obtain a purified IAL composition comprising purified IAL.
The method of any one of 2-27, wherein the separating of the halide scavenging-removal step comprises washing the first composition with a polar wash solvent to obtain the second composition.
A method of purifying an ionizable amino lipid (IAL) present in a first composition comprising the IAL and a reactive halide impurity in a nonpolar solvent, the method comprising performing a halide scavenging-removal step, comprising:
- contacting the first composition with a scavenger that reacts with the reactive halide impurity to form a scavenger-halide product;
- and
- separating the scavenger-halide product from the first composition to obtain a purified IAL composition comprising purified IAL.
A method of purifying an ionizable amino lipid (IAL) present in a composition comprising the IAL and a reactive aldehyde impurity in a first nonpolar solvent, the method comprising:
- performing an acid extraction step, comprising contacting the composition with an acid that reacts with the IAL to form an IAL salt that is soluble in a polar solvent, and transferring the IAL salt and reactive aldehyde impurity into the polar solvent to obtain an IAL salt solution;
- and
- performing an aldehyde scavenging-removal step, comprising:
- contacting the IAL salt solution with an aldehyde scavenger that reacts with the reactive aldehyde impurity to form a scavenger-aldehyde product;
- converting the IAL salt into the IAL with a neutralizing agent, wherein the IAL is soluble in a second nonpolar solvent;
- and
- separating the IAL from the scavenger-aldehyde product, or a derivative thereof formed by the neutralizing agent, by transferring the IAL into the second nonpolar solvent to obtain a purified IAL composition comprising purified IAL.
Dependencies are inferred from the claim text.