Compositions for transfecting resistant cell types
A reagent composition for delivery nucleic acid therapeutics to cells is provided. The reagent includes ionizable lipid, structural lipid, and a stabilizing agent which improves the transfection efficiency of nucleic acids, and leaves transfected cells viable. The 5 transfection reagent is effective in plasmid and mRNA delivery, and in neurons and progenitor-like cells. It may be used for ex vivo cell therapy.
1. A transfection reagent composition comprising:
35-50 Mol % of a cationic ionizable lipid, or a pharmaceutically acceptable salt thereof;
15 to 25 Mol % structural lipid;
35 to 41 Mol % sterol; and
0.5 to about 10 Mol % of a stabilizing agent, wherein the stabilizing agent is a maltoside and comprises more than one surfactant.
2. The composition of claim 1 , wherein the cationic ionizable lipid is an amino lipid or a pharmaceutically acceptable salt thereof.
3. The composition of claim 1 , wherein the stabilizing agent includes a polysorbate.
4. The composition of claim 3 , wherein the polysorbate is polysorbate (80).
5. The composition of claim 1 , wherein the stabilizing agent includes a mixture of polysorbates.
6. A transfection reagent composition comprising:
35-50 Mol % of a cationic ionizable lipid, or a pharmaceutically acceptable salt thereof;
15 to 25 Mol % structural lipid:
35 to 41 Mol % sterol; and
0.5 to about 10 Mol % of a stabilizing agent comprising a mixture of polysorbate and maltoside.
7. The composition of claim 6 , wherein the stabilizing agent is a mixture of polysorbate 80, polysorbate 20, and Tridecyl-D-maltoside.
8. The composition of claim 7 wherein the mixture is an equal ratio of polysorbate 80, polysorbate 20, and Tridecyl-D-maltoside.
9. The composition of claim 1 , wherein the cationic ionizable lipid comprises about 40 Mol % of the composition.
10. The composition of claim 1 , wherein the stabilizing agent comprises about 0.5 to 5 Mol % of the composition.
11. The composition of claim 1 , wherein the sterol is cholesterol.
12. The composition of claim 1 , further comprising a nucleic acid or a peptide.
13. The composition of claim 12 , wherein the nucleic acid is a DNA, an RNA, or a plasmid capable of expressing an RNA.
14. The composition of claim 1 , wherein cationic the ionizable lipid is selected from the group consisting of BOCHD-C3-DMA, KC2, MC3, a-D-Tocopherolsuccinoyl and DODMA, or a pharmaceutically acceptable salt thereof.
15. The composition of claim 1 , wherein the structural lipid is selected from the group consisting of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), DSPE, or DSPC.
16. The composition of claim 1 , wherein the composition exists in the form of lipid nanoparticles having a diameter of from about 15 nm to about 300 nm.
17. A method for transfecting a cell with a nucleic acid therapeutic while maintaining activity of the nucleic acid and viability of the cell, comprising contacting the cell with the composition of claim 1 .
18. The method of claim 17 , wherein the cell is a mammalian cell.
19. The method of claim 17 , wherein the cell is a human-derived cell type.