Combinations of mRNAs encoding immune modulating polypeptides and uses thereof

Patent metadata

Field	Value
Publication number	US-11344504-B1
Application number	US-202117352904-A
Country	US
Kind code	B1
Filing date	Jun 21, 2021
Priority date	May 18, 2016
Publication date	May 31, 2022
Grant date	May 31, 2022

Abstract

The disclosure relates to compositions and methods for the preparation, manufacture and therapeutic use of combinations of immunomodulatory polynucleotides (e.g., mRNAs) encoding an immune response primer polypeptide (e.g., an interleukin 23 (IL-23) polypeptide or an interleukin 36γ (IL-36-gamma) polypeptide), and an immune response co-stimulatory signal polypeptide (e.g., an OX40L polypeptide).

First claim

What is claimed is:

1. A method for treating cancer in a subject, comprising administering intratumorally to the subject a lipid nanoparticle (LNP) comprising: (i) a first mRNA comprising an open reading frame (ORF) encoding a human IL-23 polypeptide and (ii) a second mRNA comprising an ORF encoding a human IL-36 gamma polypeptide.

2. The method of claim 1 , wherein the IL-23 polypeptide comprises an IL-12p40 polypeptide operably linked, with or without a linker, to an IL-23p19 polypeptide.

3. The method of claim 2 , wherein the IL-12p40 polypeptide is operably linked with a linker to the IL-23p19 polypeptide, and wherein the linker is a Gly/Ser linker.

4. The method of claim 3 , wherein the IL-23 polypeptide comprises the amino acid sequence as set forth in SEQ ID NO: 140.

5. The method of claim 1 , wherein the IL-36 gamma polypeptide comprises an amino acid sequence selected from the group consisting of the amino acid sequence as set forth in SEQ ID NO: 10, the amino acid sequence as set forth in SEQ ID NO: 12, and the amino acid sequence as set forth in SEQ ID NO: 16.

6. A method for treating cancer in a subject, comprising administering intratumorally to the subject a lipid nanoparticle (LNP) comprising: (i) a first mRNA comprising an open reading frame (ORF) encoding a human IL-23 polypeptide, (ii) a second mRNA comprising an ORF encoding a human IL-36 gamma polypeptide, and (iii) a third mRNA comprising an ORF encoding a human OX40L polypeptide.

7. The method of claim 6 , wherein the OX40L polypeptide comprises an amino acid sequence selected from the group consisting of the amino acid sequence as set forth in SEQ ID NO: 2, and the amino acid sequence as set forth in SEQ ID NO: 21.

8. The method of claim 7 , wherein the first mRNA encodes the amino acid sequence as set forth in SEQ ID NO: 140; (ii) the second mRNA encodes the amino acid sequence as set forth in SEQ ID NO: 16; and (iii) the third mRNA encodes the amino acid sequence as set forth in SEQ ID NO: 21.

9. The method of claim 6 , wherein the first, second, and third mRNAs are formulated in the lipid nanoparticle at a mass ratio of IL-23:IL-36 gamma:OX40L of 1:2:1.

10. The method of claim 6 , wherein each of the first, second and third mRNAs comprise a 5′cap, a 5′untranslated region (UTR), a 3′UTR comprising a miR-122 binding site and a polyA tail.

11. The method of claim 6 , wherein each of the first, second and third mRNAs are fully modified with chemically-modified uridines.

12. The method of claim 1 , wherein the LNP comprises an ionizable amino lipid, a phospholipid, a sterol, and a PEG-modified lipid.

13. The method of claim 12 , wherein the LNP comprises a molar ratio of about 20-60% ionizable amino lipid, about 5-25% phospholipid, about 25-55% sterol, and about 0.5-1.5% PEG-modified lipid.

14. The method of claim 13 wherein the lipid nanoparticle comprises (i) a molar ratio of 40-60% ionizable amino lipid, 8-16% phospholipid, 30-45% sterol, and 1-5% PEG modified lipid; or (ii) a molar ratio of 45-65% ionizable amino lipid, 5-10% phospholipid, 25-40% sterol, and 0.5-5% PEG modified lipid.

15. The method of claim 14 , wherein the lipid nanoparticle comprises a molar ratio of 40-60% Compound 18, 8-16% DSPC, 30-45% cholesterol, and 1-5% PEG DMG.

16. The method of claim 14 , wherein the lipid nanoparticle comprises a molar ratio of 45-65% Compound 18, 5-10% DSPC, 25-40% cholesterol, and 0.5-5% PEG DMG.

17. The method of claim 12 , wherein the ionizable lipid comprises a compound having the formula:

18. The method of claim 12 , wherein the PEG lipid comprises PEG DMG.

19. The method of claim 12 , wherein the phospholipid comprises DSPC.

20. The method of claim 12 , wherein the structural lipid comprises cholesterol.

21. The method of claim 5 , wherein the IL-36 gamma polypeptide comprises the amino acid sequence of SEQ ID NO: 16.

22. The method of claim 2 , wherein the ORF of the first mRNA comprises from 5′ to 3′ (a) a nucleotide sequence encoding the IL-12p40 polypeptide and a nucleotide sequence encoding the IL-23p19 polypeptide, or (b) a nucleotide sequence encoding the IL-12p40 polypeptide, a nucleotide sequence encoding a peptide linker, and a nucleotide sequence encoding the IL-23p19 polypeptide.

23. The method of claim 1 , wherein each of the first and second mRNAs comprise a 5′cap, a 5′untranslated region (UTR), a 3′UTR comprising a miR-122 binding site and a polyA tail.

24. The method of claim 1 , wherein each of the first and second mRNAs are fully modified with chemically-modified uridines.

25. The method of claim 6 , wherein the IL-23 polypeptide comprises an IL-12p40 polypeptide operably linked, with or without a linker, to an IL-23p19 polypeptide.

26. The method of claim 25 , wherein the ORF of the first mRNA comprises from 5′ to 3′ (a) a nucleotide sequence encoding the IL-12p40 polypeptide and a nucleotide sequence encoding the IL-23p19 polypeptide, or (b) a nucleotide sequence encoding the IL-12p40 polypeptide, a nucleotide sequence encoding a peptide linker, and a nucleotide sequence encoding the IL-23p19 polypeptide.

27. The method of claim 6 , wherein the LNP comprises an ionizable amino lipid, a phospholipid, a sterol, and a PEG-modified lipid.

28. The method of claim 27 , wherein the LNP comprises (i) a molar ratio of about 20-60% ionizable amino lipid, about 5-25% phospholipid, about 25-55% sterol, and about 0.5-1.5% PEG-modified lipid; (ii) a molar ratio of 40-60% ionizable amino lipid, 8-16% phospholipid, 30-45% sterol, and 1-5% PEG modified lipid; or (iii) a molar ratio of 45-65% ionizable amino lipid, 5-10% phospholipid, 25-40% sterol, and 0.5-5% PEG modified lipid.

29. The method of claim 6 , wherein the lipid nanoparticle comprises (i) a molar ratio of 40-60% Compound 18, 8-16% DSPC, 30-45% cholesterol, and 1-5% PEG DMG; or (ii) a molar ratio of 45-65% Compound 18, 5-10% DSPC, 25-40% cholesterol, and 0.5-5% PEG DMG.

Assignees

• Modernatx Inc

Inventors

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

Classifications

• A61K38/177 Primary

  Receptors; Cell surface antigens; Cell surface determinants · CPC title

• A61K2039/545

  characterised by the dose, timing or administration schedule · CPC title

• A61K2039/53

  DNA (RNA) vaccination · CPC title

• A61K38/2006

  IL-1 · CPC title

• A61K2039/585

  wherein the target is cancer · CPC title