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, 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.
2. The lipid nanoparticle of claim 1 , 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.
3. The lipid nanoparticle of claim 1 , wherein each of the first mRNA, second mRNA, and third mRNA comprise a 3′ untranslated region (UTR) comprising at least one microRNA-binding site.
4. The lipid nanoparticle of claim 3 , wherein the at least one microRNA binding site is a miR-122 binding site.
5. The lipid nanoparticle of claim 4 , wherein the miR-122 binding site is a miR-122-5p binding site.
6. The lipid nanoparticle of claim 5 , wherein the miR-122-5p binding site comprises the nucleotide sequence as set forth in SEQ ID NO: 26.
7. The lipid nanoparticle of claim 3 , wherein the 3′UTR comprises a nucleotide sequence selected from the group consisting of: the nucleotide sequence as set forth in SEQ ID NO: 120, and a nucleotide sequence having at least 90% identity to SEQ ID NO: 120.
8. The lipid nanoparticle of claim 1 , wherein each of the first, second, and third mRNAs comprise a 5′cap and a poly-A tail of about 100 nucleotides in length.
9. The lipid nanoparticle of claim 1 , wherein each of the first, second, and third mRNAs comprise a 5′ untranslated region (UTR).
10. The lipid nanoparticle of claim 9 , wherein the 5′UTR comprises a nucleotide sequence selected from the group consisting of: the nucleotide sequence as set forth in SEQ ID NO: 27, and a nucleotide sequence having at least 90% identity to SEQ ID NO: 27.
11. The lipid nanoparticle of claim 1 , wherein each of the first, second, and third mRNAs are chemically modified.
12. The lipid nanoparticle of claim 11 , wherein each of the first, second, and third mRNAs are fully modified with N1-methylpseudouridine.
13. The lipid nanoparticle of claim 1 , wherein the first, second, and third mRNAs are formulated in the lipid nanoparticle at a mass ratio of OX40L:IL-23:IL-36gamma of 1:1:2.
14. The lipid nanoparticle of claim 1 , comprising an ionizable amino lipid comprising a compound having the formula:
15. The lipid nanoparticle of claim 14 , further comprising a phospholipid, a sterol, and a PEG-modified lipid.
16. 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.
17. The lipid nanoparticle of claim 16 , wherein the miR-122-5p binding site comprises the nucleotide sequence as set forth in SEQ ID NO: 26.
18. The lipid nanoparticle of claim 17 , wherein each of the first, second, and third mRNAs are fully modified with N1-methylpseudouridine.
19. The lipid nanoparticle of claim 16 , comprising an ionizable amino lipid comprising a compound having the formula:
20. The lipid nanoparticle of claim 19 , further comprising a phospholipid, a sterol, and a PEG-modified lipid.
21. 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.
22. The lipid nanoparticle of claim 21 , wherein each of the first, second, and third mRNAs are chemically modified.
23. The lipid nanoparticle of claim 22 , wherein each of the first, second, and third mRNAs are fully modified with N1-methylpseudouridine.
24. The lipid nanoparticle of claim 23 , wherein the first, second, and third mRNAs are formulated in the lipid nanoparticle at a mass ratio of OX40L:IL-23:IL-36gamma of 1:1:2.
25. The lipid nanoparticle of claim 21 , comprising an ionizable amino lipid comprising a compound having the formula:
26. The lipid nanoparticle of claim 25 , further comprising a phospholipid, a sterol, and a PEG-modified lipid.
27. A lipid nanoparticle comprising:
(i) a first mRNA comprising the nucleotide sequence of SEQ ID NO: 142;
(ii) a second mRNA comprising the nucleotide sequence of SEQ ID NO: 144; and
(iii) a third mRNA comprising the nucleotide sequence of SEQ ID NO: 146.
28. The lipid nanoparticle of claim 27 , wherein each of the first, second, and third mRNAs are fully modified with N1-methylpseudouridine.
29. The lipid nanoparticle of claim 28 , comprising an ionizable amino lipid comprising a compound having the formula:
30. The lipid nanoparticle of claim 29 , further comprising a phospholipid, a sterol, and a PEG-modified lipid.