Polynucleotides encoding interleukin-12 (IL12) and uses thereof

Patent metadata

Field	Value
Publication number	US-10646549-B2
Application number	US-201816192274-A
Country	US
Kind code	B2
Filing date	Nov 15, 2018
Priority date	May 18, 2016
Publication date	May 12, 2020
Grant date	May 12, 2020

Abstract

The present disclosure relates to polynucleotides comprising an open reading frame of linked nucleosides encoding human interleukin-12 (IL12), functional fragments thereof, and fusion proteins comprising IL12. In some embodiments, the open reading frame is sequence-optimized. In particular embodiments, the disclosure provides sequence-optimized polynucleotides comprising nucleotides encoding the polypeptide sequence of human IL12, or sequences having high sequence identity with those sequence optimized polynucleotides.

First claim

What is claimed is:

1. A method for treating cancer in a subject by inducing an anti-tumor immune response, comprising administering intratumorally to the subject a lipid nanoparticle (LNP) encapsulated messenger RNA (mRNA) comprising an open reading frame (ORF) encoding a human IL-12 polypeptide, wherein the LNP comprises a molar ratio of about 20-60% ionizable amino lipid; 5-25% phospholipid; 25-55% sterol; and 0.5-15% PEG-modified lipid.

2. The method of claim 1 , wherein the ORF encodes a human IL-12B polypeptide operably linked to a human IL-12A polypeptide.

3. The method of claim 2 , wherein the IL-12B polypeptide is operably linked to the IL-12A polypeptide by a peptide linker, and wherein the peptide linker comprises a Gly/Ser linker.

4. The method of claim 1 , wherein the human IL12 polypeptide comprises an amino acid sequence set forth in SEQ ID NO: 48.

5. The method of claim 1 , wherein the anti-tumor response comprises a T-cell response, IFNγ production, or both a T-cell response and IFNγ production.

6. A method for treating cancer in a subject by inducing an anti-tumor immune response, comprising administering intratumorally to the subject a LNP encapsulated mRNA comprising an ORF encoding a human IL-12 polypeptide, wherein the LNP comprises an ionizable amino lipid; a phospholipid; a sterol; and a PEG-modified lipid, and wherein the mRNA comprises an ORF comprising a nucleotide sequence at least 90% identical to SEQ ID NO: 237.

7. The method of claim 6 , wherein the mRNA comprises the nucleotide sequence as set forth in SEQ ID NO: 237.

8. The method of claim 7 , wherein the mRNA comprises a 3′ UTR comprising a microRNA binding site, wherein the microRNA binding site is a miR-122-3p or a miR-122-5p binding site.

9. The method of claim 8 , wherein the miR-122-5p binding site comprises the sequence set forth in SEQ ID NO: 54.

10. The method of claim 7 , wherein the mRNA comprises a 3′ UTR comprising the sequence set forth in SEQ ID NO: 240.

11. The method of claim 10 , wherein the mRNA comprises a 5′ UTR comprising the sequence set forth in SEQ ID NO: 135.

12. The method of claim 7 , wherein the mRNA is fully modified with chemically-modified uridines.

13. The method of claim 12 , wherein the chemically-modified uridines are N1-methylpseudouridines (m1ψ).

14. The method of claim 6 , wherein the anti-tumor response comprises a T-cell response, IFNγ production, or both a T-cell response and IFNγ production.

15. The method of claim 1 , wherein the mRNA comprises a nucleotide sequence at least 90% to SEQ ID NO: 96.

16. The method of claim 15 , wherein the mRNA comprises the nucleotide sequence as set forth in SEQ ID NO: 96.

17. The method of claim 16 , wherein the mRNA is fully modified with N1-methylpseudouridines (m1ψ).

18. The method of claim 1 , wherein the ionizable amino lipid comprises a compound having the formula

19. The method of claim 1 , wherein the LNP comprises a molar ratio of about 50% ionizable amino lipid; 10% phospholipid; 38.5% cholesterol; and 1.5% PEG-modified lipid.

20. The method of claim 19 , wherein the ionizable amino lipid comprises a compound having the formula

Assignees

• Modernatx Inc

Inventors

• Frederick Joshua
• Hewitt Susannah
• Bai Ailin
• Hoge Stephen
• Presnyak Vladimir
• Mcfadyen Iain
• Benenato Kerry
• Kumarasinghe Ellalahewage Sathyajith

Classifications

• A61K9/5123

  Organic compounds, e.g. fats, sugars · CPC title

• A61K38/208 Primary

  IL-12 · CPC title

• C07K14/5434

  IL-12 · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61K9/0019 Primary

  Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner (non-active ingredients are additionally classified in A61K47/00) · CPC title