In vivo production of proteins

Patent metadata

Field	Value
Publication number	US-10583203-B2
Application number	US-201916442168-A
Country	US
Kind code	B2
Filing date	Jun 14, 2019
Priority date	Apr 2, 2012
Publication date	Mar 10, 2020
Grant date	Mar 10, 2020

Abstract

The invention relates to compositions including polynucleotides encoding polypeptides which have been chemically modified by replacing the uridines with 1-methyl-pseudouridine to improve one or more of the stability and/or clearance in tissues, receptor uptake and/or kinetics, cellular access by the compositions, engagement with translational machinery, mRNA half-life, translation efficiency, immune evasion, protein production capacity, secretion efficiency, accessibility to circulation, protein half-life and/or modulation of a cell's status, function, and/or activity.

First claim

What is claimed is:

1. A method of expressing a protein in a mammalian subject, the method comprising administering a pharmaceutical composition comprising a plurality of lipid nanoparticles encapsulating a polynucleotide, wherein the lipid nanoparticle comprises a biodegradable cationic lipid, a neutral lipid, cholesterol, and a PEGylated lipid and the plurality of lipid nanoparticles has a mean particle size of between 80 nm to 160 nm, and
wherein the polynucleotide comprises:
(a) an open reading frame encoding the protein consisting of nucleotides selected from 1-methyl-pseudouridine, cytidine, adenosine, and guanosine;
(b) a 5′-UTR;
(c) at least one 5′ cap structure;
(d) a 3′-UTR; and
(e) a 3′ tailing sequence of linked nucleosides.

2. The method of claim 1 , wherein the biodegradable cationic lipid comprises an ester linkage.

3. The method of claim 1 , wherein the method comprises administering about 0.05 to about 0.5 mg/kg of polynucleotide.

4. The method of claim 1 , wherein the administration is intramuscular administration.

5. The method of claim 1 , wherein the administration is intravenous administration.

6. The method of claim 1 , wherein upon administration, expression of the protein is maximal at 8-24 hours.

7. The method of claim 1 , wherein the 3′-tailing sequence of linked nucleosides is selected from the group consisting of a poly-A tail and a polyA-G quartet.

8. The method of claim 7 , wherein the poly-A comprises approximately 160 nucleotides.

9. The method of claim 1 , wherein the at least one 5′ cap structure is selected from the group consisting of Cap0, Cap1, ARCA, inosine, N1-methyl-guanosine, 2′fluoro-guanosine, 7-deaza-guanosine, 8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine, and 2-azido-guanosine.

10. The method of claim 9 , wherein the at least one 5′-cap structure is cap0, cap1, or ARCA.

11. The method of claim 1 , wherein the plurality of lipid nanoparticles has a mean PDI of between 0.02 and 0.2.

12. The method of claim 1 , wherein the plurality of lipid nanoparticles has a mean lipid to polynucleotide ratio (wt/wt) of between 10 and 20.

13. The method of claim 1 , wherein the 3′-UTR comprises a miR binding site.

14. The method of claim 1 , wherein the 5′-UTR comprises a Kozak sequence.

15. The method of claim 1 , wherein the neutral lipid is a phospholipid.

16. The method of claim 1 , wherein the open reading frame is codon optimized to bias GC content.

17. The method of claim 1 , wherein the plurality of lipid nanoparticles comprise about 50 mol % biodegradable cationic lipid, about 38.5% cholesterol, about 10% neutral lipid and about 1.5% PEGylated lipid.

18. The method of claim 1 , wherein the polynucleotide includes at least two stop codons before the 3′ untranslated region (UTR).

Assignees

• Modernatx Inc

Inventors

• De Fougerolles Antonin
• Guild Justin

Classifications

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P37/08

  Antiallergic agents (antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14) · CPC title

• A61P37/06

  Immunosuppressants, e.g. drugs for graft rejection · CPC title

• A61P37/02

  Immunomodulators · CPC title

• A61P37/00

  Drugs for immunological or allergic disorders · CPC title