Combinations of mRNAs encoding immune modulating polypeptides and uses thereof
View Patent ↗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).
1. A lipid nanoparticle 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,
wherein the lipid nanoparticle comprises a molar ratio of 20-60% ionizable amino lipid, 5-25% phospholipid, 25-55% sterol, and 0.5-1.5% PEG-modified lipid.
2. The lipid nanoparticle of claim 1 , wherein the lipid nanoparticle comprises a molar ratio of 50% ionizable amino lipid, 10% phospholipid, 38.5% sterol, and 1.5% PEG-modified lipid.
3. The lipid nanoparticle of claim 1 , wherein the ionizable amino lipid comprises a compound having the formula:
4. The lipid nanoparticle of claim 2 , wherein the ionizable amino lipid comprises a compound having the formula:
5. The lipid nanoparticle of claim 4 , wherein the sterol is cholesterol and the PEG-modified lipid is PEG-DMG.
6. The lipid nanoparticle of claim 4 , wherein the human IL-23 polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 140; the human IL-36 gamma polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 16; and the human OX40L polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 21.
7. A lipid nanoparticle comprising:
(i) a first mRNA comprising an ORF encoding a human IL-23 polypeptide, wherein the ORF is at least 80% identical to the nucleotide sequence of SEQ ID NO: 141;
(ii) a second mRNA comprising an ORF encoding a human IL-36 gamma polypeptide, wherein the ORF is at least 80% identical to the nucleotide sequence of SEQ ID NO: 143; and
(iii) a third mRNA comprising an ORF encoding a human OX40L polypeptide, wherein the ORF is at least 80% identical to the nucleotide sequence of SEQ ID NO: 145,
wherein the lipid nanoparticle comprises a molar ratio of 20-60% ionizable amino lipid, 5-25% phospholipid, 25-55% sterol, and 0.5-1.5% PEG-modified lipid.
8. The lipid nanoparticle of claim 7 , wherein the lipid nanoparticle comprises a molar ratio of 50% ionizable amino lipid, 10% phospholipid, 38.5% sterol, and 1.5% PEG-modified lipid.
9. The lipid nanoparticle of claim 7 , wherein the ionizable amino lipid comprises a compound having the formula:
10. The lipid nanoparticle of claim 8 , wherein the ionizable amino lipid comprises a compound having the formula:
11. The lipid nanoparticle of claim 10 , wherein the sterol is cholesterol and the PEG-modified lipid is PEG-DMG.
12. The lipid nanoparticle of claim 10 , wherein
(i) the first mRNA comprises an ORF at least 85%, at least 90%, at least 95%, or at least 98% identical to the nucleotide sequence of SEQ ID NO: 141;
(ii) the second mRNA comprises an ORF at least 85%, at least 90%, at least 95%, or at least 98% identical to the nucleotide sequence of SEQ ID NO: 143; and
(iii) the third mRNA comprises an ORF at least 85%, at least 90%, at least 95%, or at least 98% identical to the nucleotide sequence of SEQ ID NO: 145.
13. The lipid nanoparticle of claim 12 , wherein each of the first mRNA, second mRNA, and third mRNA comprise a 3′ untranslated region (UTR) comprising at least one microRNA-binding site.
14. The lipid nanoparticle of claim 13 , wherein the at least one microRNA binding site is a miR-122 binding site.
15. The lipid nanoparticle of claim 14 , wherein the miR-122 binding site is a miR-122-5p binding site.
16. The lipid nanoparticle of claim 15 , wherein the miR-122-5p binding site comprises the nucleotide sequence as set forth in SEQ ID NO: 26.
17. The lipid nanoparticle of claim 16 , wherein each of the first, second, and third mRNAs are chemically modified.
18. The lipid nanoparticle of claim 17 , wherein each of the first, second, and third mRNAs are fully modified with N1-methylpseudouridine.
19. A lipid nanoparticle, comprising:
(i) a first mRNA comprising an ORF comprising the nucleotide sequence of SEQ ID NO: 141;
(ii) a second mRNA comprising an ORF comprising the nucleotide sequence of SEQ ID NO: 143; and
(iii) a third mRNA comprising an ORF comprising the nucleotide sequence of SEQ ID NO: 145,
wherein each of the first mRNA, second mRNA, and third mRNA comprise a 3′ UTR comprising at least one miR-122-5p microRNA-binding site, and wherein the lipid nanoparticle comprises a molar ratio of 20-60% ionizable amino lipid, 5-25% phospholipid, 25-55% sterol, and 0.5-1.5% PEG-modified lipid.
20. The lipid nanoparticle of claim 19 , wherein the lipid nanoparticle comprises a molar ratio of 50% ionizable amino lipid, 10% phospholipid, 38.5% sterol, and 1.5% PEG-modified lipid.
21. The lipid nanoparticle of claim 19 , wherein the ionizable amino lipid comprises a compound having the formula:
22. The lipid nanoparticle of claim 20 , wherein the ionizable amino lipid comprises a compound having the formula:
23. The lipid nanoparticle of claim 22 , wherein the sterol is cholesterol and the PEG-modified lipid is PEG-DMG.
24. The lipid nanoparticle of claim 21 , wherein the miR-122-5p binding site comprises the nucleotide sequence as set forth in SEQ ID NO: 26.
25. The lipid nanoparticle of claim 24 , wherein each of the first, second, and third mRNAs are chemically modified.
26. The lipid nanoparticle of claim 25 , wherein each of the first, second, and third mRNAs are fully modified with N1-methylpseudouridine.
27. A lipid nanoparticle comprising
(i) a first mRNA encoding a human IL-23 polypeptide, wherein the first mRNA comprises a nucleotide sequence at least 90% identical to the nucleotide sequence of SEQ ID NO: 142;
(ii) a second mRNA encoding a human IL-36 gamma polypeptide, wherein the second mRNA comprises a nucleotide sequence at least 90% identical to the nucleotide sequence of SEQ ID NO: 144; and
(iii) a third mRNA encoding a human OX40L polypeptide, wherein the third mRNA comprises a nucleotide sequence at least 90% identical to the nucleotide sequence of SEQ ID NO: 146,
wherein the lipid nanoparticle comprises a molar ratio of 50% ionizable amino lipid, 10% phospholipid, 38.5% sterol, and 1.5% PEG-modified lipid.
28. The lipid nanoparticle of claim 27 , wherein the ionizable amino lipid comprises a compound having the formula:
29. The lipid nanoparticle of claim 28 , wherein
(i) the first mRNA comprises the nucleotide sequence of SEQ ID NO: 142;
(ii) the second mRNA comprises the nucleotide sequence of SEQ ID NO: 144; and
(iii) the third mRNA comprises the nucleotide sequence of SEQ ID NO: 146.
30. The lipid nanoparticle of claim 29 , wherein each of the first, second, and third mRNAs are fully modified with N1-methylpseudouridine.