Modified nucleosides, nucleotides, and nucleic acids, and uses thereof

Patent metadata

Field	Value
Publication number	US-10064959-B2
Application number	US-201715493829-A
Country	US
Kind code	B2
Filing date	Apr 21, 2017
Priority date	Oct 1, 2010
Publication date	Sep 4, 2018
Grant date	Sep 4, 2018

Abstract

The present disclosure provides modified nucleosides, nucleotides, and nucleic acids, and methods of using thereof.

First claim

What is claimed is:

1. A synthetic messenger ribonucleic acid (mRNA) that is synthesized according to a method comprising the steps of:
a) providing a complementary deoxyribonucleic acid (cDNA) that encodes a pharmaceutical protein of interest;
b) selecting a nucleotide that disrupts a binding of a major groove binding partner with the mRNA, wherein the nucleotide has decreased binding affinity to the major groove binding partner selected from the group consisting of toll-like receptor (TLR) 3, TLR7, TLR8, retinoic acid-inducible gene I (RIG-I), melanoma differentiation-associated gene 5 (MDA5) and laboratory of genetics and physiology 2 (LGP2), and wherein the nucleotide comprises a modification on the major groove face of the nucleobase where an atom of the major groove face of the nucleobase is replaced or substituted with an alkyl group; and
c) contacting the provided cDNA and the selected nucleotide with an RNA polymerase under conditions such that an mRNA transcript is synthesized.

2. The synthetic mRNA of claim 1 , wherein the method further comprises after c), the step of:
d) 5′-capping the mRNA transcript concomitantly or post-transcriptionally such that the mRNA is synthesized.

3. The synthetic mRNA of claim 1 , wherein the mRNA is at least 300 nucleotides in length.

4. The synthetic mRNA of claim 1 , wherein the nucleotide comprises a modification on the major groove face of a pyrimidine nucleobase.

5. The synthetic mRNA of claim 4 , wherein the pyrimidine nucleobase is selected from cytosine and uracil.

6. The synthetic mRNA of claim 5 , wherein the pyrimidine nucleobase is uracil.

7. The synthetic mRNA of claim 5 , wherein the pyrimidine nucleobase is cytosine.

8. The synthetic mRNA of claim 5 , wherein the nucleotide comprises 1-methyl-pseudouridine or 5-methyl-uridine.

9. The synthetic mRNA of claim 8 , wherein the nucleotide comprises 1-methyl-pseudouridine.

10. The synthetic mRNA of claim 7 , wherein the nucleotide comprises 5-methyl-cytidine.

11. The synthetic mRNA of claim 4 , wherein the major groove binding partner is TLR3, TLR7, or TLR8.

12. The synthetic mRNA of claim 4 , wherein the major groove binding partner is RIG-I, MDA5, or LGP2.

13. A pharmaceutical composition comprising the synthetic mRNA of claim 1 and a pharmaceutically acceptable carrier.

14. A pharmaceutical composition comprising the synthetic mRNA of claim 3 and a pharmaceutically acceptable carrier.

15. A pharmaceutical composition comprising the synthetic mRNA of claim 4 and a pharmaceutically acceptable carrier.

16. A pharmaceutical composition comprising the synthetic mRNA of claim 6 and a pharmaceutically acceptable carrier.

17. A pharmaceutical composition comprising the synthetic mRNA of claim 7 and a pharmaceutically acceptable carrier.

18. A pharmaceutical composition comprising the synthetic mRNA of claim 8 and a pharmaceutically acceptable carrier.

19. A pharmaceutical composition comprising the synthetic mRNA of claim 9 and a pharmaceutically acceptable carrier.

20. A pharmaceutical composition comprising the synthetic mRNA of claim 10 and a pharmaceutically acceptable carrier.

21. The synthetic mRNA of claim 2 , wherein the 5′-capping is performed concomitantly.

22. The synthetic mRNA of claim 2 , wherein the mRNA is at least 300 nucleotides in length.

23. The synthetic mRNA of claim 2 , wherein the nucleotide comprises a modification on the major groove face of a pyrimidine nucleobase.

24. The synthetic mRNA of claim 23 , wherein the pyrimidine nucleobase is selected from cytosine and uracil.

25. The synthetic mRNA of claim 24 , wherein the nucleotide comprises 1-methyl-pseudouridine or 5-methyl-uridine.

26. The synthetic mRNA of claim 23 , wherein the major groove binding partner is TLR3, TLR7, or TLR8.

27. The synthetic mRNA of claim 23 , wherein the major groove binding partner is RIG-I, MDA5, or LGP2.

28. A pharmaceutical composition comprising the synthetic mRNA of claim 2 and a pharmaceutically acceptable carrier.

29. A pharmaceutical composition comprising the synthetic mRNA of claim 22 and a pharmaceutically acceptable carrier.

30. A pharmaceutical composition comprising the synthetic mRNA of claim 23 and a pharmaceutically acceptable carrier.

Assignees

• Modernatx Inc

Inventors

• Schrum Jason P
• Siddiqi Suhaib
• Ejebe Kenechi

Classifications

• C12N15/102

  Mutagenizing nucleic acids · CPC title

• C12N15/1138

  against receptors or cell surface proteins · CPC title

• G01N33/559

  through a gel, e.g. Ouchterlony technique · CPC title

• C07H19/10

  with the saccharide radical esterified by phosphoric or polyphosphoric acids · CPC title

• C07K16/00

  Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies · CPC title