Lipid formulations for delivery of messenger RNA

Patent metadata

Field	Value
Publication number	US-11890377-B2
Application number	US-202117162656-A
Country	US
Kind code	B2
Filing date	Jan 29, 2021
Priority date	Oct 22, 2013
Publication date	Feb 6, 2024
Grant date	Feb 6, 2024

Abstract

The present invention provides, among other things, methods of delivering mRNA in vivo, including administering to a subject in need of delivery a composition comprising an mRNA encoding a protein, encapsulated within a liposome such that the administering of the composition results in the expression of the protein encoded by the mRNA in vivo, wherein the liposome comprises a cationic lipid of formula I-c:

or a pharmaceutically acceptable salt thereof.

First claim

We claim:

1. A method of delivery of messenger RNA (mRNA) in vivo, comprising administering to a subject in need of delivery a composition comprising an mRNA that encodes Factor IX (FIX), wherein said mRNA is encapsulated within a liposome such that the administering of the composition results in the expression of the protein encoded by the mRNA in vivo; and
wherein the liposome comprises a cationic lipid of formula I-g:

or a pharmaceutically acceptable salt thereof,
wherein:
R L is C 1-50 alkyl, C 2-50 alkenyl, or C 2-50 alkynyl, and
wherein the mRNA has a length of or greater than about 0.5 kb.

2. The method of claim 1 , wherein R L is C 8-12 alkyl.

3. The method of claim 2 , wherein R L is linear —C 8 H 17 , —C 9 H 19 , —C 10 H 21 , —C 11 H 23 , or —C 12 H 25 .

4. The method of claim 3 , wherein R L is linear —C 8 H 17 or —C 10 H 21 .

5. The method of claim 4 , wherein the cationic lipid is cKK-E12:

6. The method of claim 1 , wherein the liposome further comprises one or more non-cationic lipids, one or more cholesterol-based lipids and/or one or more PEG-modified lipids.

7. The method of claim 6 , wherein the one or more non-cationic lipids are selected from DSPC (1,2-distearoyl-sn-glycero-3-phosphocholine), DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine), DOPE (1,2-dioleyl-sn-glycero-3-phosphoethanolamine), DOPC (1,2-dioleyl-sn-glycero-3-phosphotidylcholine) DPPE (1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine), DMPE (1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine), DOPG (1,2-dioleoyl-sn-glycero-3-phospho-(1′-rac-glycerol)).

8. The method of claim 6 , wherein the one or more cholesterol-based lipids are cholesterol and/or PEGylated cholesterol.

9. The method of claim 6 , wherein the one or more PEG-modified lipids comprise a poly(ethylene) glycol chain of up to 5 kDa in length covalently attached to a lipid with alkyl chain(s) of C 6 -C 20 length.

10. The method of claim 6 , wherein the liposome comprises cKK-E12, DOPE, cholesterol and DMG-PEG2K.

11. The method of claim 1 , wherein the cationic lipid constitutes about 30-50% of the liposome by molar ratio.

12. The method of claim 11 , wherein the ratio of cKK-E12:DOPE:cholesterol:DMG-PEG2K is approximately 40:30:20:10, 40:30:25:5, or 40:32:25:3 by molar ratio.

13. The method of claim 1 , wherein the liposome has a size less than about 250 nm, 200 nm, 150 nm, 100 nm, 75 nm, or 50 nm.

14. The method of claim 1 , wherein the expression of the protein encoded by the mRNA is detectable in liver, kidney, heart, spleen, serum, brain, skeletal muscle, lymph nodes, skin, and cerebrospinal fluid.

15. The method of claim 1 , wherein the expression of the protein encoded by the mRNA is detectable 3, 6, 12, or 24 hours after the administration or 1 week after the administration.

16. The method of claim 1 , wherein the mRNA has a length of or greater than about 1 kb, 1.5 kb, 2 kb, 2.5 kb, 3 kb, 3.5 kb, 4 kb, 4.5 kb, or 5 kb.

17. The method of claim 1 , wherein the mRNA comprises one or more modified nucleotides.

18. The method of claim 17 , wherein the one or more modified nucleotides comprise pseudouridine, N-1-methyl-pseudouridine, 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyl adenosine, 5-methylcytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5-propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, 0(6)-methylguanine, and/or 2-thiocytidine.

19. The method of claim 1 , wherein the mRNA is unmodified.

20. The method of claim 1 , wherein the mRNA encodes a therapeutic protein.

Assignees

• Translate Bio Inc
• Massachusetts Inst Technology

Inventors

• Heartlein Michael
• Anderson Daniel
• Dong Yizhou
• Derosa Frank

Classifications

• A61K9/1272 Primary

  comprising non-phosphatidyl surfactants as bilayer-forming substances, e.g. cationic lipids or non-phosphatidyl liposomes coated or grafted with polymers (lipids as modifying agents {A61K47/543}) · CPC title

• A61K38/177

  Receptors; Cell surface antigens; Cell surface determinants · CPC title

• A61K38/1709

  from mammals · CPC title

• A61K38/4846

  Factor VII (3.4.21.21); Factor IX (3.4.21.22); Factor Xa (3.4.21.6); Factor XI (3.4.21.27); Factor XII (3.4.21.38) · CPC title

• A61K38/53

  Ligases (6) · CPC title