Combinations of mRNAs encoding immune modulating polypeptides and uses thereof

Patent metadata

Field	Value
Publication number	US-11071716-B2
Application number	US-202017071685-A
Country	US
Kind code	B2
Filing date	Oct 15, 2020
Priority date	May 18, 2016
Publication date	Jul 27, 2021
Grant date	Jul 27, 2021

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 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 fusion protein comprising a human IL-12p40 polypeptide fused directly or by a linker to a human IL-23p19 polypeptide, wherein the IL-12p40 polypeptide comprises the amino acid sequence of SEQ ID NO: 3, wherein the IL-23p19 polypeptide comprises the amino acid sequence of SEQ ID NO: 5, wherein the LNP comprises an ionizable amino lipid; a phospholipid; a sterol; and a PEG-modified lipid, and wherein the mRNA encoded fusion protein induces the anti-tumor immune response thereby treating cancer.

2. The method of claim 1 , wherein the IL-12p40 polypeptide is fused directly to the IL-23p19 polypeptide without a linker.

3. The method of claim 1 , wherein the IL-12p40 polypeptide is fused directly to the IL-23p19 polypeptide with a peptide linker.

4. The method of claim 1 , wherein the fusion protein comprises a non-native signal peptide.

5. The method of claim 1 , wherein the fusion protein comprises a human IL-12p40 signal peptide or a human IL-23p19 signal peptide.

6. The method of claim 1 , wherein the human IL-12p40 polypeptide comprises the amino acid sequence of SEQ ID NO: 1.

7. The method of claim 1 , wherein the ORF comprises from 5′ to 3′ a nucleotide sequence selected from the group consisting of:
(i) a nucleotide sequence encoding the IL-12p40 polypeptide, a nucleotide sequence encoding a peptide linker, and a nucleotide sequence encoding the IL-23p19 polypeptide;
(ii) a nucleotide sequence encoding the IL-12p40 polypeptide, and a nucleotide sequence encoding the IL-23p19 polypeptide;
(iii) a nucleotide sequence encoding the IL-23p19 polypeptide, a nucleotide sequence encoding a peptide linker, and a nucleotide sequence encoding the IL-12p40 polypeptide; and
(iv) a nucleotide sequence encoding the IL-23p19 polypeptide, and a nucleotide sequence encoding the IL-12p40 polypeptide.

8. The method of claim 1 , wherein the ORF comprises a nucleotide sequence encoding a signal peptide located at the 5′ terminus of the ORF.

9. The method of claim 1 , wherein the mRNA comprises (i) a 3′ UTR; (ii) a 5′ UTR and (iii) a polyA tail.

10. The method of claim 1 , wherein the mRNA comprises a chemically modified nucleotide.

11. 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 fusion protein, wherein the ORF comprises from 5′ to 3′:
(i) a nucleotide sequence encoding a signal peptide;
(ii) a nucleotide sequence encoding a human IL-12p40 polypeptide comprising the amino acid sequence of SEQ ID NO: 3;
(iii) a nucleotide sequence encoding a peptide linker;
(iv) a nucleotide sequence encoding a human IL-23p19 polypeptide comprising the amino acid sequence of SEQ ID NO: 5,
wherein the LNP comprises an ionizable amino lipid; a phospholipid; a sterol; and a PEG-modified lipid, and wherein the mRNA encoded fusion protein induces an anti-tumor immune response thereby treating cancer.

12. The method of claim 11 , wherein the signal peptide is selected from: (i) a non-native signal peptide; (ii) a human IL-23p19 signal peptide; and (iii) a human IL-12p40 signal peptide.

13. The method of claim 11 , wherein the human IL-12p40 polypeptide comprises the amino acid sequence of SEQ ID NO: 1.

14. The method of claim 11 , wherein the mRNA comprises (i) a 3′ UTR; (ii) a 5′ UTR and (iii) a polyA tail.

15. The method of claim 11 , wherein the mRNA comprises a chemically modified nucleotide.

16. 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 fusion protein, wherein the ORF comprises from 5′ to 3′:
(i) a nucleotide sequence encoding a signal peptide;
(ii) a nucleotide sequence encoding a human IL-12p40 polypeptide comprising the amino acid sequence of SEQ ID NO: 3;
(iii) a nucleotide sequence encoding a human IL-23p19 polypeptide comprising the amino acid sequence of SEQ ID NO: 5,
wherein the LNP comprises an ionizable amino lipid; a phospholipid; a sterol; and a PEG-modified lipid, and wherein the mRNA encoded fusion protein induces an anti-tumor immune response thereby treating cancer.

17. The method of claim 16 , wherein the signal peptide is selected from: (i) a non-native signal peptide; (ii) a human IL-23p19 signal peptide; and (iii) a human IL-12p40 signal peptide.

18. The method of claim 16 , wherein the human IL-12p40 polypeptide comprises the amino acid sequence of SEQ ID NO: 1.

19. The method of claim 16 , wherein the mRNA comprises (i) a 3′ UTR; (ii) a 5′ UTR and (iii) a polyA tail.

20. The method of claim 16 , wherein the mRNA comprises a chemically modified nucleotide.

Assignees

• Modernatx Inc

Inventors

• Frederick Joshua
• 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

• A61K31/7115

  Nucleic acids or oligonucleotides having modified bases, i.e. other than adenine, guanine, cytosine, uracil or thymine · CPC title

• A61K2039/545

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

• C07K14/5443

  IL-15 · CPC title

• C07K14/5434

  IL-12 · CPC title