MRNA Therapies Including SIRP-ALPHA
Human SIRPa fusion proteins having an enhanced affinity to CD47 via increased binding valency. The human SIR-Pa fusion proteins described herein may form tetramer, hexamer, octamers, etc. These fusion proteins may be configured to form heterodimers with other fusion proteins, including C-C chemokine receptor type 4 (CCR4) binding fusion proteins.
1 . An mRNA polynucleotide sequence encoding for a fusion protein, wherein the fusion protein comprises:
a) a first signal regulatory protein alpha (SIRPα) polypeptide that binds to a CD47 protein;
b) an IgG Fc domain, wherein the IqG Fc domain comprises at least one modification that facilitates heterodimerization of the fusion protein with another fusion protein that comprises an IqG Fc domain; and
c) a second SIRPα polypeptide that binds to the CD47 protein,
wherein the IqG Fc domain is linked to the first SIRPα polypeptide or the second SIRPα polypeptide via a linker, wherein the first SIRPα polypeptide and the second SIRPα polypeptide are not directly linked to each other.
2 . The mRNA polynucleotide of claim 1 , wherein the IgG Fc domain comprises a human IqG1 Fc domain, wherein the human IqG1 Fc domain comprises a CH2 domain and a CH3 domain.
3 . The mRNA polynucleotide of claim 1 , wherein the IgG Fc domain comprises a knob modification and/or a hole modification.
4 . The mRNA polynucleotide of claim 1 , wherein the fusion protein comprises an amino acid sequence selected from the group consisting of; SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6.
5 . The mRNA polynucleotide of claim 1 , wherein the fusion protein comprises;
an amino acid sequence that is at least 95% homologous to the amino acid sequence of any one of SEQ ID NO: 2, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 19 or SEQ ID NO: 20.
6 . The mRNA polynucleotide of claim 1 , wherein the fusion protein comprises:
a sequence comprising amino acids 23 to 627 of SEQ ID NO: 35, a sequence comprising amino acids 23 to 617 of SEQ ID NO: 59, or a sequence comprising amino acids 23 to 742 of SEQ ID NO: 60.
7 . The mRNA polynucleotide of claim 1 , wherein the mRNA polynucleotide comprises: the nucleotide sequence of any one of SEQ ID NO: 54, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 56, SEQ ID NO: 55, SEQ ID NO: 58, SEQ ID NO: 53, SEQ ID NO: 51, SEQ ID NO: 29, SEQ ID NO: 49, SEQ ID NO: 57, or SEQ ID NO: 50.
8 . The mRNA polynucleotide of claim 1 , wherein the fusion protein is a bivalent fusion protein comprising a human IgG1 Fc domain, wherein the first SIRPα polypeptide is linked to the human IqG1 Fc domain via a first linker and the second SIRPα Polypeptide is linked to the same human IqG1 Fc domain via a second linker.
9 . The mRNA polynucleotide of claim 1 , wherein the fusion protein is a multivalent fusion protein comprising three, four, five, six, seven, or eight independent SIRPα polypeptides and a human IgG1 Fc domain.
10 . The mRNA polynucleotide of claim 1 , wherein the fusion protein is a tetravalent fusion protein comprising four independent SIRPα polypeptides and a human IgG1 Fc domain.
11 . The mRNA polynucleotide of claim 1 , wherein the fusion protein comprises;
a dimer of an amino acid sequence that is at least 95% homologous to the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, a sequence comprising amino acids 23 to 372 of SEQ ID NO: 29, a sequence comprising amino acids 23 to 601 of SEQ ID NO: 30, a sequence comprising amino acids 23 to 727 of SEQ ID NO: 31, a sequence comprising amino acids 23 to 497 of SEQ ID NO: 34, a sequence comprising amino acids 23 to 627 of SEQ ID NO: 35, a sequence comprising amino acids 23 to 492 of SEQ ID NO: 36, or a combination thereof.
12 . The mRNA polynucleotide of claim 10 , wherein the fusion protein comprises;
a dimer of an amino acid sequence of any one of SEQ ID NO: 2, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, a sequence comprising amino acids 23 to 372 of SEQ ID NO: 29, a sequence comprising amino acids 23 to 601 of SEQ ID NO: 30, a sequence comprising amino acids 23 to 727 of SEQ ID NO: 31, a sequence comprising amino acids 23 to 497 of SEQ ID NO: 34, a sequence comprising amino acids 23 to 627 of SEQ ID NO: 35, a sequence comprising amino acids 23 to 492 of SEQ ID NO: 36, or a combination thereof.
13 . The mRNA polynucleotide of claim 10 , wherein the mRNA polynucleotide comprises the nucleotide sequence of any one of SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 51, SEQ ID NO: 49, SEQ ID NO: 57, SEQ ID NO: 50, or SEQ ID NO: 65.
14 . The mRNA polynucleotide of claim 1 , wherein the fusion protein is a hexavalent fusion protein comprising six independent SIRPα polypeptides and a human IgG1 Fc domain.
15 . The mRNA polynucleotide of claim 14 , wherein the fusion protein comprises;
a dimer of an amino acid sequence that is at least 95% homologous to the amino acid sequence of any one of SEQ ID NO: 12, a sequence comprising amino acids 23 to 627 of SEQ ID NO: 35, a sequence comprising amino acids 23 to 617 of SEQ ID NO: 59, or a combination thereof.
16 . The mRNA polynucleotide of claim 14 , wherein the fusion protein comprises:
a dimer of an amino acid sequence any one of SEQ ID NO: 12, a sequence comprising amino acids 23 to 627 of SEQ ID NO: 35, a sequence comprising amino acids 23 to 617 of SEQ ID NO: 59, or a combination thereof.
17 . The mRNA polynucleotide of claim 14 , wherein the mRNA polynucleotide comprises the nucleotide sequence of any one of SEQ ID NO: 56 or SEQ ID NO: 62.
18 . The mRNA polynucleotide of claim 1 , wherein the fusion protein is an octavalent fusion protein comprising eight independent SIRPα domains and a human IgG1 Fc domain.
19 .- 27 . (canceled)
28 . A fusion protein encoded by the mRNA polynucleotide of claim 1 .
29 . A therapeutic composition comprising the mRNA polynucleotide of claim 1 ; and a delivery vehicle molecule comprising an amino-lipidated peptoid.
30 .- 70 . (canceled)