Method of synthesis of an ionizable cationic lipid
What is described is a method of synthesis of the compound of formula 1A, or a salt thereof, wherein R 3 is a linear or branched alkene of 1, 2, 3, 4, 5 or 6 carbons; R 4 and R 5 are the same or different, each a hydrogen, or a linear or branched alkyl of 1, 2, 3, 4, 5 or 6 carbons; and L 3 is a bond or an alkane of 1, 2, 3, 4, 5 or 6 carbons.
1. A method of delivering a nucleic acid into a hepatocyte, comprising:
administering a composition comprising the nucleic acid encapsulated in a lipid formulation, the lipid formulation comprising:
a.) a compound of Formula IA, or a pharmaceutically acceptable salt thereof:
wherein;
R 3 is a linear or branched alkylene of 1 to 6 carbons;
R 4 and R 5 are the same or different, each, independently, a hydrogen or a linear or branched alkyl of 1 to 6 carbons; and
L 3 is a bond or an alkylene of 1 to 6 carbons;
b.) a helper lipid; and
c.) a cholesterol.
2. The method of claim 1 , wherein the nucleic acid is an RNA.
3. The method of claim 2 , wherein the RNA is siRNA, mRNA, or miRNA.
4. The method of claim 1 , wherein the lipid formulation is a lipid nanoparticle or a liposome.
5. The method of claim 4 , wherein 30-70% of the lipids in the lipid formulation are a compound having Formula IA.
6. The method of claim 5 , wherein 5-30% of the lipids in the lipid formulation are a helper lipid.
7. The method of claim 5 , wherein 20-40% of the lipids in the lipid formulation are cholesterol.
8. The method of claim 1 , wherein the composition is administered in an aqueous solution to a subject via inhalation.
9. The method of claim 8 , wherein the composition is administered to the subject via pulmonary inhalation.
10. The method of claim 8 , wherein the aqueous solution is aerosolized, atomized, or nebulized.
11. The method of claim 1 , wherein the compound of Formula IA is selected from the group consisting of
or a pharmaceutically acceptable salt thereof.
12. The method of claim 11 , wherein the compound of Formula IA is
or a pharmaceutically acceptable salt thereof.
13. The method of claim 11 , wherein the compound of Formula IA is
or a pharmaceutically acceptable salt thereof.
14. The method of claim 1 , wherein the nucleic acid is a mRNA and delivery of the mRNA into the hepatocyte results in expression of the mRNA into a protein of interest.
15. The method of claim 14 , wherein the protein of interest is an enzyme.
16. The method of claim 14 , wherein the protein of interest is an antigen.
17. The method of claim 1 , wherein the nucleic acid is an siRNA and delivery of the siRNA into the hepatocyte results in reduced expression of a gene of interest.
18. The method of claim 1 , wherein the nucleic acid comprises one or more modified nucleotides selected from phosphorothioates, phosphoramidates, methyl phosphonates, 2′-O-methyl ribonucleotides, and peptide-nucleic acids.