Novel neuropilin/growth factor complexes and uses thereof
AssigneesLudwig Institute for Cancer Research, Licentia Ltd.
InventorsKari Alitalo, Ulf Eriksson, Brigitta Olofsson, Taija Makinen
Life of this patent
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Abstract
Complexes of a protein selected from the group consisting of VEGF-B167, VEGF-C, VEGF-D and processed VEGF-B186 and analogs thereof and the neuropilin-1 (NP-1) receptor, the extracellular domain or a ligand-binding fragment or analogue thereof; the use of such complexes in assays for growth factor proteins having substantially the same binding affinity for a cell surface receptor as VEGF-B167, VEGF-C, VEGF-D or processed VEGF-B186 and/or in promoting or antagonizing a cellular response mediated by VEGF-B167, VEGF-C, VEGF-D and/or processed VEGF-B186; and specific binding partners, e.g. antibodies, for such complexes.
Claims21
An isolated ligand-receptor complex comprising two molecules, one of said molecules defining said ligand and comprising the amino acid sequence encoded by exon 6B of VEGF-B167 or a receptor-binding analog thereof, and a second of said molecules defining said receptor and being selected from the group consisting of a polypeptide chain comprising the amino acid sequence of NP-1, an extracellular domain thereof or a ligand-binding fragment thereof, a VEGF-B167-binding or VEGF-C-binding receptor analog thereof having at least 90% amino acid identity with said ligand-binding fragment, and a polypeptide chain having binding affinity for VEGF-B167 or VEGF-C and encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid encoding said ligand-binding fragment.
An isolated ligand-receptor complex comprising two molecules, one of said molecules defining said ligand and being selected from the group consisting of VEGF-B167, processed VEGF-B186, VEGF-C, and VEGF-D and a second of said molecules defining said receptor and being selected from the group consisting of a polypeptide chain comprising the amino acid sequence of NP-1, an extracellular domain thereof or a ligand-binding fragment thereof, a VEGF-B167-binding, processed VEGF-B186-binding or VEGF-C-binding receptor analog thereof having at least 90% amino acid identity with said ligand-binding fragment, and a polypeptide chain having binding affinity for VEGF-B167, processed VEGF-B186 or VEGF-C and encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid encoding said ligand-binding fragment.
A soluble fusion protein comprising an immunoglobulin and a receptor moiety selected from the group consisting of a polypeptide chain comprising the amino acid sequence of NP-1, an extracellular domain thereof or a ligand-binding fragment thereof, a VEGF-B167-binding, processed VEGF-B186-binding or VEGF-C-binding receptor analog thereof having at least 90% amino acid identity with said ligand-binding fragment, and a polypeptide chain having binding affinity for VEGF-B167, processed VEGF-B186 or VEGF-C and encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid encoding said ligand-binding fragment.
A modular assembly comprising at least a first protein domain and a second protein domain, said first domain comprising the amino acid sequence encoded by exon 6B of VEGF-B167, and said second domain being selected from VEGF165, VEGF-C, VEGF-D and processed VEGF-B186.
A method of modulating binding and bioactivity of a growth factor selected from the group consisting of VEGF-B167, VEGF-C, VEGF-D and processed VEGF-B186, relative to living cells, said method comprising introducing into said cells a vector comprising a nucleotide sequence encoding a receptor sequence selected from the group consisting of a polypeptide chain comprising the amino acid sequence of NP-1, an extracellular domain thereof or a ligand-binding fragment thereof, a VEGF-B167-binding, processed VEGF-B186-binding or VEGF-C-binding receptor analog thereof having at least 90% amino acid identity with said ligand-binding fragment, and a polypeptide chain having binding affinity for VEGF-B167, processed VEGF-B186 or VEGF-C and encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid said ligand-binding fragment.
A method of inhibiting bioactivity of a growth factor selected from the group consisting of VEGF-B167, VEGF-C, VEGF-D and processed VEGF-B186, relative to living cells, said method comprising treating said cells with a polypeptide comprising the amino acid sequence encoded by exon 6B of VEGF-B167 or a polypeptide comprising an NP-1 binding sequence encoded by exon 6A of VEGF-B186.
A method of inhibiting bioactivity of a growth factor selected from the group consisting of VEGF-B167, VEGF-C, VEGF-D and processed VEGF-B186, relative to living cells, said method comprising treating said cells with a polypeptide comprising the NP-1 binding epitope encoded by exon 6B of VEGF-B167 or a polypeptide comprising an NP-1 binding sequence encoded by exon 6A of VEGF-B186.
A method of modulating the responsiveness of cells to collapsin/semaphorin glycoproteins, said method comprising treating said cells with a substance selected from the group consisting of VEGF-B167, VEGF-C, VEGF-D and processed VEGF-B186.
A method of modulating the responsiveness of cells to collapsin/semaphorin glycoproteins, said method comprising treating said cells with a polypeptide comprising the NP-1 binding epitope encoded by exon 6B of VEGF-B167 or a polypeptide comprising an NP-1 binding sequence encoded by exon 6A of VEGF-B186.
A method of modulating the binding and bioactivity of a protein selected from the group consisting of VEGF-B167, VEGF-C, VEGF-D and processed VEGF-B186, relative to living cells, said method comprising introducing into cells a vector comprising a nucleotide sequence encoding a receptor sequence selected from the group consisting of a polypeptide chain comprising the amino acid sequence of NP-1, an extracellular domain thereof or a ligand-binding fragment thereof, a VEGF-B167-binding, processed VEGF-B186-binding or VEGF-C-binding receptor analog thereof having at least 90% amino acid identity with said ligand-binding fragment, and a polypeptide chain having binding affinity for VEGF-B167, processed VEGF-B186 or VEGF-C and encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid encoding said ligand-binding fragment, whereby said cells are caused to express said receptor sequence.
A method of inhibiting growth or proliferation of tumor cells comprising the steps of treating said cells with:
- (a) a soluble protein selected from the group consisting of a polypeptide chain comprising the amino acid sequence of NP-1, an extracellular domain thereof or a ligand-binding fragment thereof, a VEGF-B167-binding, processed VEGF-B186-binding or VEGF-C-binding receptor analog thereof having at least 90% amino acid identity with said ligand-binding fragment, and a polypeptide chain having binding affinity for VEGF-B167, processed VEGF-B186 or VEGF-C and encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid encoding said ligand-binding fragment;
- or
- (b) a polypeptide comprising the NP-1 binding epitope encoded by exon 6B of VEGF-B167 or the NP-1 binding epitope encoded by exon 6A of processed VEGF-B186.
An isolated binding partner having specific binding affinity for an epitope on a ligand-receptor complex, said complex comprising a protein selected from the group consisting of VEGF-B167, VEGF-C, VEGF-D and processed VEGF-B186 and analogs thereof in specific binding interaction with the ligand binding domain of a protein selected from the group consisting of a polypeptide chain comprising the amino acid sequence of NP-1, an extracellular domain thereof or a ligand-binding fragment thereof, a VEGF-B167-binding, processed VEGF-B186-binding or VEGF-C-binding receptor analog thereof having at least 90% amino acid identity with said ligand-binding fragment, and a polypeptide chain having binding affinity for VEGF-B167, processed VEGF-B186 or VEGF-C and encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid encoding said ligand-binding fragment;
- said binding partner having substantially no binding affinity for uncomplexed VEGF-B167, VEGF-C, VEGF-D, processed VEGF-B186 or analog thereof.
Dependencies are inferred from the claim text.