Terminally Modified RNA

Patent metadata

Field	Value
Publication number	US-10925935-B2
Application number	US-201816152945-A
Country	US
Kind code	B2
Filing date	Oct 5, 2018
Priority date	Nov 26, 2012
Publication date	Feb 23, 2021
Grant date	Feb 23, 2021

Abstract

The invention relates to compositions and methods for the manufacture and optimization of modified mRNA molecules via optimization of their terminal architecture.

First claim

The invention claimed is:

1. A lipid nanoparticle comprising an mRNA, wherein the mRNA comprises
(a) a 5′ untranslated region (5′UTR);
(b) a region of linked nucleosides encoding a polypeptide of interest;
(c) a 3′ untranslated region (3′UTR) comprising at least one microRNA binding site; and
(d) a 3′ tailing region of linked nucleosides;

and wherein each uridine in the mRNA is a modified uridine nucleoside.

2. The lipid nanoparticle of claim 1 , wherein each uridine in the mRNA is a pseudouridine analog.

3. The lipid nanoparticle of claim 2 , wherein the pseudouridine analog is 1-methyl pseudouridine.

4. The lipid nanoparticle of claim 3 , wherein each cytidine in the mRNA is a 5-methyl cytidine.

5. The lipid nanoparticle of claim 3 , wherein the microRNA binding site is for an immune cell specific microRNA.

6. The lipid nanoparticle of claim 5 , wherein the microRNA binding site is for a microRNA selected from miR-142-3p, miR-142-5p, miR-146a and miR-146b.

7. The lipid nanoparticle of claim 6 , wherein the microRNA binding site is miR-142-3p.

8. The lipid nanoparticle of claim 7 , comprising a cationic or ionizable lipid.

9. The lipid nanoparticle of claim 8 , wherein the cationic lipid is DLin-MC3-DMA, DLin-DMA, C12-200, or DLin-KC2-DMA.

10. The lipid nanoparticle of claim 8 , comprising a PEG lipid.

11. The lipid nanoparticle of claim 10 , comprising 1-5% PEG lipid.

12. The lipid nanoparticle of claim 1 , wherein the polypeptide of interest is a therapeutic protein, cytokine, growth factor, antibody or a fusion protein.

13. The lipid nanoparticle of claim 12 , wherein the region encoding the polypeptide of interest is codon optimized.

14. The lipid nanoparticle of claim 1 , wherein the mRNA further comprises a 5′ cap structure.

15. The lipid nanoparticle of claim 14 , wherein the 5′ cap structure is Cap 1.

16. The lipid nanoparticle of claim 1 , wherein the 5′ UTR comprises a translation initiation sequence selected from a group consisting of a Kozak sequence and an internal ribosome entry site (IRES).

17. The lipid nanoparticle of claim 1 , wherein the 3′ tailing region comprises a poly A tail of at least 100 nucleosides.

18. The lipid nanoparticle of claim 1 , wherein the microRNA binding site is for a microRNA selected from miR-122, miR-133, miR-206, miR-208, miR-17-92, miR-126, miR-142-3p, miR-142-5p, miR-16, miR-21, miR-223, miR-24, miR-27, let-7, miR-30c, miR-1d, miR-149, miR-192, miR-194, and miR-204.

19. The lipid nanoparticle of claim 3 , wherein the microRNA binding site is for a microRNA selected from miR-122, miR-133, miR-206, miR-208, miR-17-92, miR-126, miR-142-3p, miR-142-5p, miR-16, miR-21, miR-223, miR-24, miR-27, let-7, miR-30c, miR-1d, miR-149, miR-192, miR-194, and miR-204.

20. The lipid nanoparticle of claim 1 , wherein the microRNA binding site is for miR-142-3p.

21. A lipid nanoparticle comprising an mRNA, wherein the mRNA comprises
(a) a 5′ untranslated region (5′UTR);
(b) a region of linked nucleosides encoding a polypeptide of interest;
(c) a 3′ untranslated region (3′UTR) comprising at least one miR-142-3p binding site; and
(d) a 3′ tailing region of linked nucleosides;

and wherein each uridine in the mRNA is a modified uridine nucleoside.

22. The lipid nanoparticle of claim 21 , comprising a cationic or ionizable lipid.

23. The lipid nanoparticle of claim 22 , comprising a PEG lipid.

24. The lipid nanoparticle of claim 23 , comprising 1-5% PEG lipid.

25. The lipid nanoparticle of claim 21 , wherein the polypeptide of interest is a therapeutic protein, cytokine, growth factor, antibody or a fusion protein.

26. A lipid nanoparticle comprising an mRNA, wherein the mRNA comprises
(a) a 5′ untranslated region (5′UTR);
(b) a region of linked nucleosides encoding a polypeptide of interest;
(c) a 3′ untranslated region (3′UTR) comprising at least one miR-126-3p binding site; and
(d) a 3′ tailing region of linked nucleosides;

and wherein each uridine in the mRNA is a modified uridine nucleoside.

27. The lipid nanoparticle of claim 26 , comprising a cationic or ionizable lipid.

28. The lipid nanoparticle of claim 27 , comprising a PEG lipid.

29. The lipid nanoparticle of claim 28 , comprising 1-5% PEG lipid.

30. The lipid nanoparticle of claim 26 , wherein the polypeptide of interest is a therapeutic protein, cytokine, growth factor, antibody or a fusion protein.

31. A lipid nanoparticle comprising an mRNA, wherein the mRNA comprises
(a) a 5′ untranslated region (5′UTR);
(b) a region of linked nucleosides encoding a polypeptide of interest;
(c) a 3′ untranslated region (3′UTR) comprising at least one miR-122 binding site; and
(d) a 3′ tailing region of linked nucleosides;

and wherein each uridine in the mRNA is a modified uridine nucleoside.

32. The lipid nanoparticle of claim 31 , comprising a cationic or ionizable lipid.

33. The lipid nanoparticle of claim 32 , comprising a PEG lipid.

34. The lipid nanoparticle of claim 33 , comprising 1-5% PEG lipid.

35. The lipid nanoparticle of claim 31 , wherein the polypeptide of interest is a therapeutic protein, cytokine, growth factor, antibody or a fusion protein.

36. The lipid nanoparticle of claim 31 , wherein the miR-122 binding site is a miR-122-3p binding site.

37. The lipid nanoparticle of claim 31 , wherein the miR-122 binding site is a miR-122-5p binding site.

Assignees

• Moderna Tx Inc
• Modernatx Inc

Inventors

• Chakraborty Tirtha
• Bancel Stephane
• Hoge Stephen G
• Roy Atanu
• De Fougerolles Antonin
• Afeyan Noubar B

Classifications

• C12N15/67 Primary

  General methods for enhancing the expression · CPC title

• A61K39/00 Primary

  Medicinal preparations containing antigens or antibodies (materials for immunoassay G01N33/53) · CPC title

• A61P37/06

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