Granted Patent B2

US 12,409,137 · App. 17/637,278 · Granted Sep 9, 2025

Inventors: Maria Helena Christine Van Rooijen (Cambridge, MA); Hok Hei Tam (Newton, MA); Maier Steve Avendaño Amado (Cambridge, MA); Barry Andrew Martin (Boston, MA); Ignacio Martinez (Lexington, MA); Piotr Stanislaw Kowalski (Cork, IE); Nataliya Vladimirovna Nukolova (Cambridge, MA); John Patrick Casey, Jr. (Boston, MA); Siddharth Dilipkumar Patel (Cambridge, MA); Roman Lvovitch Bogorad (Newton, MA)

Assignee: Flagship Pioneering Innovations VI, LLC

A61K9/1272A61K47/60C12N15/88

Abstract

Claims (15)

1. A method for making lipid reconstructed plant messenger packs (LPMPs), the method comprising reconstituting a film comprising purified lipids derived from a plant source in the presence of a synthetic charged lipid, thereby producing a LPMP that comprises the synthetic charged lipid, wherein the synthetic charged lipid is chosen from 1′-((2-(4-(2-((2-(bis(2-hydroxydodecyl)amino)ethyl) (2-hydroxydodecyl)amino)ethyl)piperazin-1-yl)ethyl)azanediyl)bis(dodecan-2-ol) (C12-200), DLin-MC3-DMA (MC3), dioleoyl-3-trimethylammonium propane (DODAP), DC-cholesterol, DOTAP, Ethyl PC, GL67, DLin-KC2-DMA (KC2), MD1 cKK-E12 (MD1), OF2, EPC, ZA3-Ep10, TT3, LP01, 5A2-SC8, heptadecan-9-yl 8-((2-hydroxyethyl)(8-(nonyloxy)-8-oxooctyl)amino)octanoate, a cationic sulfonamide amino lipid, an amphiphilic zwitterionic amino lipid, DODAC, DOBAQ, YSK05, DOBAT, DOBAQ, DOPAT, DOMPAQ, DOAAQ, DMAP-BLP, DLinDMA, DODMA, DOTMA, DSDMA, DOSPA, DODAC, DOBAQ, DMRIE, DOTAP-cholesterol, GL67A, and 98N12-5, or combinations thereof.

2. The method of claim 1 , wherein the synthetic charged lipid is chosen from C12-200, MC3, DODAP, DC-cholesterol, DOTAP, Ethyl PC, GL67, KC2, MD1, OF2, EPC, ZA3-Ep10, TT3, LP01, 5A2-SC8, heptadecan-9-yl 8-((2-hydroxyethyl)(8-(nonyloxy)-8-oxooctyl)amino)octanoate, a cationic sulfonamide amino lipid, and an amphiphilic zwitterionic amino lipid, or combinations thereof.

3. The method of claim 2 , wherein the synthetic charged lipid is chosen from C12-200, MC3, DODAP, and DC-cholesterol, or combinations thereof.

4. The method of claim 1 , wherein the reconstitution is performed in the presence of a sterol, thereby producing a LPMP that comprises a synthetic charged lipid and a sterol.

5. The method of claim 4 , wherein the sterol is cholesterol or sitosterol.

6. The method of claim 1 , wherein the reconstitution is performed in the presence of a PEGylated lipid, thereby producing a LPMP that comprises a synthetic charged lipid and a PEGylated lipid.

7. The method of claim 6 , wherein the PEGylated lipid is C14-PEG2k, C18-PEG2k, or DMPE-PEG2k.

8. The method of claim 1 , wherein the reconstitution is performed in the presence of a sterol and a PEGylated lipid, thereby producing a LPMP that comprises a synthetic charged lipid, a sterol, and a PEGylated lipid.

9. The method of claim 8 , wherein the synthetic charged lipid, lipids derived from a plant source, sterol, and PEGylated lipid comprise a molar ratio of about 30%-75%, about 10%-20%, about 35%-50%, and about 1%-3%, respectively, of the lipids in the LPMP.

10. The method of claim 9 , wherein the synthetic charged lipid, lipids derived from a plant source, sterol, and PEGylated lipid are formulated at a molar ratio of 50:10:38.5:1.5.

11. The method of claim 1 , wherein the LPMPs comprise a heterologous polynucleotide.

12. The method of claim 11 , wherein the heterologous polynucleotide is an mRNA, a siRNA or a precursor thereof, a microRNA (miRNA) or a precursor thereof, a viral RNA, or a plasmid.

13. The method of claim 11 , wherein the encapsulation efficiency of the heterologous polynucleotide by the LPMP is at least 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or more than 99%.

14. The method of claim 1 , wherein the LPMP is formulated for delivery to an animal.

15. The method of claim 1 , wherein the LPMP is formulated for delivery to a plant.

Assignments (4)

Continuity (5)

References Cited (12)