Granted Patent B2

US 12,023,371 · App. 17/127,435 · Granted Jul 2, 2024

Inventors: Tirtha Chakraborty (Medford, MA); Stephane Bancel (Cambridge, MA); Stephen G. Hoge (Brookline, MA); Atanu Roy (Stoneham, MA); Antonin De Fougerolles (Waterloo, BE); Noubar B. Afeyan (Cambridge, MA)

Assignee: ModernaTX, Inc.

A61K39/00C12N15/67

Abstract

Claims (28)

1. A method for reducing expression of a therapeutic polypeptide in human cells or human tissue, comprising contacting the cells or tissue with a messenger RNA (mRNA) comprising:

a. a 5′ untranslated region (UTR);

b. a region of linked nucleosides encoding the therapeutic polypeptide;

c. a 3′ UTR comprising at least one microRNA binding site which binds to at least one microRNA molecule differentially expressed in the cells or tissue; and

d. a 3′ tailing region of linked nucleosides

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

wherein expression of the therapeutic polypeptide is reduced in the cells or tissue expressing the microRNA molecule.

2. The method of claim 1 , wherein the modified uridine nucleoside is a 1-methyl-pseudouridine.

3. The method of claim 1 , wherein each cytidine in the mRNA is a 5-methyl cytidine.

4. The method of claim 1 , wherein the microRNA binding site comprises a sequence selected from the group consisting of: SEQ ID Nos: 1192-2212 and 3234-4254.

5. The method of claim 1 , wherein the microRNA binding site comprises a sequence selected from the group consisting of: SEQ ID Nos: 1250, 1255, 1308, 1311, 1374, 1375, 1404 and 1405.

6. The method of claim 1 , wherein the contacting occurs in vivo.

7. The method of claim 1 , wherein the UTR 3′ UTR comprises 1, 2, 3, 4 or 5 microRNA binding sites.

8. A method for targeting expression of a therapeutic polypeptide in cells or tissue, comprising administering a lipid nanoparticle (LNP) encapsulated mRNA to a human subject, wherein the mRNA comprises:

a. a 5′ UTR;

b. a region of linked nucleosides encoding the therapeutic polypeptide;

c. a 3′ UTR comprising at least one microRNA binding site which binds to at least one microRNA molecule differentially expressed in the cells or tissue; and

d. a 3′ tailing region of linked nucleosides

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

wherein expression of the therapeutic polypeptide is reduced in the cells or tissue expressing the microRNA molecule.

9. The method of claim 8 , wherein the modified uridine nucleoside is a 1-methyl-pseudouridine.

10. The method of claim 8 , wherein each cytidine in the mRNA is a 5-methyl cytidine.

11. The method of claim 8 , wherein the microRNA binding site comprises a sequence selected from the group consisting of: SEQ ID Nos: 1192-2212 and 3234-4254.

12. The method of claim 8 , wherein the microRNA binding site comprises a sequence selected from the group consisting of: SEQ ID Nos: 1250, 1255, 1308, 1311, 1374, 1375, 1404 and 1405.

13. The method of claim 8 , wherein the UTR 3′ UTR comprises 1, 2, 3, 4 or 5 microRNA binding sites.

14. The method of claim 8 , wherein the LNP comprises a cationic or ionizable lipid.

15. The method of claim 14 , wherein the LNP comprises a PEG lipid.

16. The method of claim 15 , wherein the LNP comprises 1-5% PEG-lipid.

Assignments (3)

Continuity (14)