POLYNUCLEOTIDES ENCODING METHYLMALONYL-CoA MUTASE

• US 20190022019A1
• Filed: 08/27/2018
• Published: 01/24/2019
• Est. Priority Date: 12/17/2015
• Status: Abandoned Application

• Current Assignee: Moderna TX, Inc. (Moderna, Inc.)
• Original Assignee: Moderna TX, Inc. (Moderna, Inc.)
• Inventors: Martini, Paolo, Presnyak, Vladimir
• Application Number:
  US16/113,814
• Publication Number:
  US 20190022019A1
• Time in Patent Office: Days
• 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

1-15. -15. (canceled)

16. A messenger RNA (mRNA) comprising:

(i) a 5′

-terminal cap;

(ii) a 5′

untranslated region (UTR);

(iii) an open reading frame (ORF) encoding the human methylmalonyl-CoA mutase (MCM) polypeptide of SEQ ID NO;

208, SEQ ID NO;

209, SEQ ID NO;

210, SEQ ID NO;

211, SEQ ID NO;

212, or SEQ ID NO;

213, wherein at least 95% of uracils in the ORF are 5-methoxyuracils, and wherein the uracil content in the ORF is below 150% of the theoretical minimum;

(iv) a 3′

UTR; and

(v) a poly-A tail.

17. The mRNA of claim 16, wherein the uracil content in the ORF is above 100% of the theoretical minimum.

18. The mRNA of claim 16, wherein the uracil content in the ORF is between 123% and 125%, between 122% and 126%, between 121% and 127%, between 120% and 128%, between 119% and 129%, between 118% and 130%, between 117% and 131%, between 116% and 132%, between 115% and 133%, between 114% and 134%, between 113% and 135%, between 112% and 136%, or between 111% and 137% of the theoretical minimum.

19. The mRNA of claim 16, wherein the uracil content in the ORF is between 118% and 129% of the theoretical minimum.

20. The mRNA of claim 16, wherein the ORF encodes the human MCM polypeptide of SEQ ID NO:

213.

21. The mRNA of claim 20, wherein the ORF is at least 80% identical to the nucleotide sequence of SEQ ID NO:

732.

22. The mRNA of claim 20, wherein the ORF is at least 85% identical to the nucleotide sequence of SEQ ID NO:

732.

23. The mRNA of claim 20, wherein the ORF is at least 90% identical to the nucleotide sequence of SEQ ID NO:

732.

24. The mRNA of claim 20, wherein the ORF is at least 95% identical to the nucleotide sequence of SEQ ID NO:

732.

25. The mRNA of claim 20, wherein the ORF is at least 98% identical to the nucleotide sequence of SEQ ID NO:

732.

26. The mRNA of claim 20, wherein the ORF is at least 99% identical to the nucleotide sequence of SEQ ID NO:

732.

27. The mRNA of claim 20, wherein the ORF is 100% identical to the nucleotide sequence of SEQ ID NO:

732.

28. The mRNA of claim 21, wherein the 5′

UTR comprises the nucleotide sequence of SEQ ID NO;

215.

29. The mRNA of claim 21, wherein the mRNA comprises the miR-142-3p binding site depicted in SEQ ID NO:

722.

30. The mRNA of claim 21, wherein the 5′

terminal cap is Cap1.

31. The mRNA of claim 21, wherein the poly-A tail is 100 residues in length.

32. The mRNA of claim 27, wherein the 5′

UTR comprises the nucleotide sequence of SEQ ID NO;

215.

33. The mRNA of claim 27, wherein the mRNA comprises the miR-142-3p binding site depicted in SEQ ID NO:

722.

34. The mRNA of claim 27, wherein the 5′

terminal cap is Cap1.

35. The mRNA of claim 27, wherein the poly-A tail is 100 residues in length.

36. The mRNA of claim 27, wherein the 5′

UTR comprises the nucleotide sequence of SEQ ID NO;

215, wherein the mRNA comprises the miR-142-3p binding site depicted in SEQ ID NO;

722, wherein the 5′

terminal cap is Cap1, and wherein the poly-A tail is 100 residues in length.

37. A pharmaceutical composition comprising the mRNA of claim 16 and a pharmaceutically acceptable excipient.

38. A pharmaceutical composition comprising the mRNA of claim 27 and a pharmaceutically acceptable excipient.

39. A pharmaceutical composition comprising the mRNA of claim 36 and a pharmaceutically acceptable excipient.

40. A lipid nanoparticle comprising the mRNA of claim 16.

41. A lipid nanoparticle comprising the mRNA of claim 27.

42. A lipid nanoparticle comprising the mRNA of claim 36.

43. A method of treating methylmalonic acidemia in a human subject in need thereof, the method comprising administering to the human subject an effective amount of the pharmaceutical composition of claim 37.

44. A method of treating methylmalonic acidemia in a human subject in need thereof, the method comprising administering to the human subject an effective amount of the pharmaceutical composition of claim 38.

45. A method of treating methylmalonic acidemia in a human subject in need thereof, the method comprising administering to the human subject an effective amount of the pharmaceutical composition of claim 39.