Alternative nucleic acid molecules and uses thereof

Patent metadata

Field	Value
Publication number	US-10286086-B2
Application number	US-201515319840-A
Country	US
Kind code	B2
Filing date	Jun 19, 2015
Priority date	Jun 19, 2014
Publication date	May 14, 2019
Grant date	May 14, 2019

Abstract

The present disclosure provides alternative nucleosides, nucleotides, and nucleic acids, and methods of using them.

First claim

The invention claimed is:

1. An mRNA encoding a polypeptide, wherein at least one base is 1-methyl-pseudouracil and one base is 5-methoxy-uracil.

2. A pharmaceutical composition comprising the mRNA of claim 1 and a pharmaceutically acceptable excipient.

3. A method of expressing a polypeptide in a mammalian cell, said method comprising the steps of:
(i) providing an mRNA of claim 1 ; and
(ii) introducing said mRNA to a mammalian cell under conditions that permit the expression of the polypeptide by the mammalian cell,

thereby expressing the polypeptide in the mammalian cell.

4. The mRNA of claim 1 , wherein about 25% of the uracils are 1-methyl-pseudouracil and about 75% of the uracils are 5-methoxy-uracil.

5. The mRNA of claim 1 , wherein about 50% of the uracils are 1-methyl-pseudouracil and about 50% of the uracils are 5- methoxy-uracil.

6. The mRNA of claim 1 , wherein about 75% of the uracils are 1-methyl-pseudouracil and about 25% of the uracils are 5-methoxy-uracil.

7. The mRNA of claim 1 , wherein at least one base is 5-methyl-cytosine.

8. The mRNA of claim 1 , wherein at least one nucleotide is alpha-thio-adenosine or alpha-thio-guanosine.

9. The mRNA of claim 8 , wherein about 2-100% of the adenosines and/or guanosines in the mRNA are alpha-thio-adenosine and/or alpha-thio-guanosine.

10. The mRNA of claim 9 , wherein about 2% of the adenosines and/or guanosines in the mRNA are alpha-thio-adenosine and/or alpha-thio-guanosine.

11. The mRNA of claim 1 further comprising:
(i) at least one 5′-cap structure;
(ii) a 5′-UTR optionally comprising a Kozak sequence; and
(iii) a 3′-UTR.

12. The mRNA of claim 11 , wherein the at least one 5′-cap structure is Cap0, Cap1, ARCA, inosine, N1-methyl-guanosine, 2′-fluoro-guanosine, 7-deaza-guanosine, 8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine, or 2-azido-guanosine.

13. The mRNA of claim 11 further comprising a poly-A tail.

14. The mRNA of claim 1 , wherein the mRNA is codon optimized.

Assignees

• Modernatx Inc

Inventors

• Roy Atanu
• Conlee Christopher R
• De Fougerolles Antonin
• Fraley Andrew W
• Butora Gabor
• Stanton Matthew

Classifications

• C07D239/54

  as doubly bound oxygen atoms or as unsubstituted hydroxy radicals · CPC title

• C07D473/16

  two nitrogen atoms · CPC title

• C07D473/40

  with halogen atoms or perhalogeno-alkyl radicals directly attached in position 2 or 6 · CPC title

• C07H19/067

  with ribosyl as the saccharide radical · CPC title

• C07D487/16

  Peri-condensed systems · CPC title