Polynucleotides encoding methylmalonyl-CoA mutase

• US 10,406,112 B2
• Filed: 06/07/2018
• Issued: 09/10/2019
• Est. Priority Date: 12/17/2015
• Status: Active Grant

• Current Assignee: Moderna TX, Inc. (Moderna, Inc.)
• Original Assignee: Moderna TX, Inc. (Moderna, Inc.)
• Inventors: Martini, Paolo, Presnyak, Vladimir, Benenato, Kerry
• Primary Examiner(s): Shin, Dana H
• Application Number:
  US16/002,376
• Publication Number:
  US 20180271795A1
• Time in Patent Office: 460 Days
• Field of Search: None
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 38/52 Isomerases (5)
• CPC Class Codes: A61K 48/005 characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
• CPC Class Codes: A61K 9/1272 with substantial amounts of non-phosphatidyl, i.e. non-acylglycerophosphate, surfactants as bilayer-forming substances, e.g. cationic lipids with cholesterol as the only non-phosphatidyl surfactant A61K9/127; lipids as modifying agent A61K47/543
• CPC Class Codes: A61K 9/5123 Organic compounds, e.g. fats, sugars
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 15/52 Genes encoding for enzymes or proenzymes
• CPC Class Codes: C12N 9/90 Isomerases (5.)
• CPC Class Codes: C12Y 504/99002 Methylmalonyl-CoA mutase (5.4.99.2)

Abstract

The disclosure relates to polynucleotides comprising an open reading frame of linked nucleosides encoding human methylmalonyl-CoA mutase precursor, human methylmalonyl-CoA mutase (MCM) mature form, or functional fragments thereof. In some embodiments, the disclosure includes methods of treating methylmalonic acidemia in a subject in need thereof comprising administering an mRNA encoding an MCM polypeptide.

Claims

1. A lipid nanoparticle comprising a messenger RNA (mRNA) and a compound having the formula:

2. The lipid nanoparticle of claim 1, wherein the ORF encodes the human MCM polypeptide of SEQ ID NO:

213.

3. The lipid nanoparticle of claim 2, wherein the ORF is at least 99% identical to the nucleotide sequence of SEQ ID NO:

732.

4. The lipid nanoparticle of claim 2, wherein the ORF is 100% identical to the nucleotide sequence of SEQ ID NO:

732.

5. The lipid nanoparticle of claim 2, wherein the mRNA comprises a 5′

untranslated region (UTR) comprising the nucleotide sequence of SEQ ID NO;

215.

6. The lipid nanoparticle of claim 2, wherein the mRNA comprises a 5′

terminal cap comprising a guanine cap nucleotide containing an N7 methylation and the 5′

-terminal nucleotide of the mRNA contains a 2′

-O-methyl.

7. The lipid nanoparticle of claim 2, wherein the mRNA comprises a poly-A tail 100 residues in length.

8. The lipid nanoparticle of claim 2, wherein at least 95% of uridines in the mRNA are 5-methoxyuridines.

9. The lipid nanoparticle of claim 4, wherein the mRNA comprises a 5′

UTR comprising the nucleotide sequence of SEQ ID NO;

215.

10. The lipid nanoparticle of claim 4, wherein the mRNA comprises a 5′

terminal cap comprising a guanine cap nucleotide containing an N7 methylation and the 5′

-terminal nucleotide of the mRNA contains a 2′

-O-methyl.

11. The lipid nanoparticle of claim 4, wherein the mRNA comprises a poly-A tail 100 residues in length.

12. The lipid nanoparticle of claim 4, wherein at least 95% of uridines in the mRNA are 5-methoxyuridines.

13. The lipid nanoparticle of claim 4, wherein the mRNA comprises a 5′

UTR comprising the nucleotide sequence of SEQ ID NO;

215, wherein the mRNA comprises a 5′

terminal cap comprising a guanine cap nucleotide containing an N7 methylation and the 5′

-terminal nucleotide of the mRNA contains a 2′

-O-methyl, wherein the mRNA comprises a poly-A tail 100 residues in length, and wherein at least 95% of uridines in the mRNA are 5-methoxyuridines.

14. The lipid nanoparticle of claim 1, further comprising a phospholipid, a structural lipid, and a PEG lipid.

15. The lipid nanoparticle of claim 4, further comprising a phospholipid, a structural lipid, and a PEG lipid.

16. The lipid nanoparticle of claim 13, further comprising a phospholipid, a structural lipid, and a PEG lipid.

17. The lipid nanoparticle of claim 14, wherein the phospholipid is 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) and the structural lipid is cholesterol.

18. The lipid nanoparticle of claim 15, wherein the phospholipid is 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) and the structural lipid is cholesterol.

19. The lipid nanoparticle of claim 16, wherein the phospholipid is 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) and the structural lipid is cholesterol.

20. The lipid nanoparticle of claim 1, wherein the ORF encodes the human MCM polypeptide of SEQ ID NO:

208.

21. The lipid nanoparticle of claim 1, wherein the ORF encodes the human MCM polypeptide of SEQ ID NO:

209.

22. The lipid nanoparticle of claim 1, wherein the ORF encodes the human MCM polypeptide of SEQ ID NO:

210.

23. The lipid nanoparticle of claim 1, wherein the ORF encodes the human MCM polypeptide of SEQ ID NO:

211.

24. The lipid nanoparticle of claim 1, wherein the ORF encodes the human MCM polypeptide of SEQ ID NO:

212.