Terminally modified RNA

Patent metadata

Field	Value
Publication number	US-10155029-B2
Application number	US-201715429532-A
Country	US
Kind code	B2
Filing date	Feb 10, 2017
Priority date	Nov 26, 2012
Publication date	Dec 18, 2018
Grant date	Dec 18, 2018

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 mRNA comprising
(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,
wherein each uridine in the mRNA is a modified uridine nucleoside.

2. The mRNA of claim 1 , wherein the modified uridine nucleoside is a pseudouridine analog.

3. The mRNA of claim 2 , wherein the pseudouridine analog is a 1-methyl pseudouridine.

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

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

6. The synthetic isolated terminally optimized mRNA of claim 1 , comprising at least one 5′ cap structure.

7. The mRNA of claim 6 , 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, 2-azido-guanosine, Cap2, Cap4, and CAP-003-CAP-225.

8. The mRNA of claim 1 , wherein the at least one microRNA binding site is for an immune cell specific microRNA.

9. The mRNA of claim 8 , wherein the immune cell specific microRNA is selected from the group consisting of miR-142-3p, miR-142-5p, miR-146a and miR-146b.

10. The mRNA of claim 1 , wherein the 3′ tailing region of linked nucleosides further comprises a chain terminating nucleoside.

11. The mRNA of claim 10 , wherein the chain terminating nucleoside is selected from the group consisting of 3′-deoxyadenosine (cordycepin), 3′-deoxyuridine, 3′-deoxycytosine, 3′-deoxyguanosine, 3′-deoxythymine, 2′,3′-dideoxynucleosides, 2′,3′-dideoxyadenosine, 2′,3′-dideoxyuridine, 2′,3′-dideoxycytosine, 2′,3′-dideoxyguanosine, 2′,3′-dideoxythymine, a 2′-deoxynucleoside, and —O— methylnucleoside.

12. The mRNA of claim 1 , wherein the 3′ tailing region comprises a stem loop sequence.

13. The mRNA of claim 1 , wherein the modified uridine nucleoside is 1-methyl pseudouridine, and wherein each cytidine in the mRNA is 5-methyl cytidine.

14. The mRNA of claim 1 , wherein the 3′ tailing region of linked nucleosides comprises a poly A tail of at least 100, at least 120 or at least 140 nucleosides.

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

Assignees

• Modernatx Inc

Inventors

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

Classifications

• A61P37/06

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

• 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