IgM and IgE heavy chain domain 2 as covalently linked homodimerization modules for the generation of fusion proteins with dual specificity
The present invention provides a polypeptides comprising a heavy chain domain 2 (HD2) from IgM or IgE and at least one pharmaceutically active moiety, complexes thereof and their use for therapy and prophylaxis.
1. A polypeptide comprising a heavy chain domain 2 (EHD2) from IgE and at least one pharmaceutically active moiety, under the proviso that the pharmaceutically active moiety is not a Fab or Fc fragment from IgM or IgE, and wherein the polypeptide does not comprise an IgE CH3 domain or CH4 domain,
wherein the EHD2 domain comprises the amino acid sequence according to SEQ ID NO: 2 or is a dimerizing variant thereof having at least 95% sequence identity to SEQ ID NO: 2, and
wherein the pharmaceutically active moiety is selected from the group consisting of an anti-EGFR scFv comprising the amino acid sequence of SEQ ID NO:4, an anti-HER2 scFv comprising the amino acid sequence of SEQ ID NO:3, and a TNF-related apoptosis-inducing ligand (TRAIL) comprising the amino acid sequence of SEQ ID NO:6.
2. The polypeptide of claim 1 , wherein at least one pharmaceutically active moiety is connected to the N- and/or C-Terminus of the EHD2.
3. The polypeptide of claim 1 , wherein at least one pharmaceutically active moiety is connected to the EHD2 directly or indirectly via one or more linkers.
4. The polypeptide of claim 3 , wherein the one or more linkers comprise peptide linkers.
5. The polypeptide of claim 4 , wherein the one or more linkers comprise one or more cleavage sites.
6. The polypeptide of claim 1 , wherein the polypeptide comprises at least two pharmaceutically active moieties connected to the EHD2, wherein the at least two pharmaceutically active moieties are identical pharmaceutically active moieties or are different pharmaceutically active moieties are.
7. The polypeptide of claim 1 , wherein at least one pharmaceutically active moiety is selected from the group consisting of a half-life extension module, and an imaging molecule.
8. The polypeptide of claim 7 , wherein the at least one pharmaceutically active moiety is a half-life extension module, wherein the half-life extension module is selected from the group consisting of immunoglobulin binding domains (IgBD), albumin, albumin-binding domains (ABD), peptides, small molecules, fatty acids, antibody fragments, single-domain antibodies, VHH, scaffold proteins, and natural ligands exhibiting affinity for a long-circulating plasma protein, which are optionally PEGylated, HESylated, Polysialylated, N-glycosylated, O-glycosylated, or PEG- mimicking polypeptides.
9. The polypeptide of claim 7 , wherein the at least one pharmaceutically active moiety is an imaging molecule, wherein the imaging molecule is selected from the group consisting of bioluminescent reagents, chemiluminescent reagents, fluorescent imaging reagents, photosensitizers, chelating reagents, and radioactive moieties.
10. A complex comprising at least two polypeptides according to claim 1 .
11. The complex of claim 10 , wherein the at least two polypeptides are connected via their EHD2 domains.
12. The complex of claim 10 , wherein the at least two polypeptides are connected via covalent or non-covalent bonds.
13. The complex of claim 12 , wherein the covalent bond is a disulfide bond.
14. The complex of claim 10 , wherein the at least two polypeptides are identical or different.
15. A pharmaceutical composition comprising the polypeptide of claim 1 .
16. The pharmaceutical composition of claim 15 , further comprising a pharmaceutically acceptable carrier and/or excipient.
17. The polypeptide of claim 1 , wherein the polypeptide comprises an amino acid sequence selected from the group of SEQ ID NO: 15, 16, 17, 29, 32, and 33.
18. A polypeptide comprising an amino acid sequence selected from the group of SEQ ID NO: 15, 16, 17, 28, 29, 31, 32, and 33.