# US 7,407,766 B1 · Screening method for agonists or antagonists of semaphorin 6C by determining the effect of a test compound on the interaction between semaphorin 6C and plexin-A1 > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 7,407,766 B1 - Document ID: US7407766B1 - Status: Granted - Kind: B1 - Application number: 10/148,409 - Filed: 2000-11-27 - Granted: 2008-08-05 - Earliest claimed: 1999-11-30 - Assignees: Japan Science and Technology Agency; Dainippon Sumitomo Pharma Co., Ltd. - Inventors: Hajime Fujisawa; Toru Kimura; Kaoru Kikuchi; Yasunori Murakami - Web page: https://patentgrep.com/patent/US7407766B1 ## CPC - G01N 33/5008: {for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics} - A61P 25/02: for peripheral neuropathies - A61P 27/02: Ophthalmic agents - A61P 27/16: Otologicals - A61P 35/00: Antineoplastic agents - A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 - C07K 14/47: from mammals - C07K 14/4703: {Inhibitors; Suppressors} - C07K 14/705: Receptors; Cell surface antigens; Cell surface determinants {(tumour specific antigens C07K14/4748)} - C07K 14/71: for growth factors; for growth regulators - G01N 33/502: {for testing non-proliferative effects} - G01N 33/68: involving proteins, peptides or amino acids {(involving lipoproteins G01N33/92)} - G01N 33/6896: {Neurological disorders, e.g. Alzheimer's disease} - C07K 2319/00: Fusion polypeptide - G01N 2333/4704: Inhibitors; Supressors - G01N 2500/00: Screening for compounds of potential therapeutic value - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) ## Abstract The present invention provides screening methods for agonists or antagonists of Sema6C using Plexin-A1, and tools for the screening and the like. It evaluates a case of making a recombinant protein having an extracellular domain of Semaphorin 6C contact a protein having an extracellular domain of Plexin-A1 by comparing it with a case of contacting a recombinant protein and a target substance having an extracellular domain of Semaphorin 6C, or the target substance with a protein having an extracellular domain of Plexin-A1. The evaluation will be carried out by the connectivity of a protein having an extracellular domain of Semaphorin 6C, a growth cone collapse activity of Plexin-A1 expressing cell, or a contractile activity of Plexin-A1 expressing cell. ## Claims ### Claim 1 A screening method for agonists or antagonists of Semaphorin 6C, comprising the steps of: a) measuring in an in vitro assay system the binding activity of a protein with an extracellular domain of Plexin-A1, comprising the 1st to 1237th amino acid residues in the sequence of Plexin-A1, consisting of SEQ ID NO:5 or SEQ ID NO:6, to a protein having an extracellular domain of Semaphorin 6C, comprising the 1st to 599th amino acid residues in the sequence of Semaphorin 6C, consisting of SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10, in the presence and absence of a target substance, and b) selecting the target substance as a candidate for an agonist or antagonist of Semaphorin 6C if there is a decrease in said binding activity in the presence of said target substance compared to an assay in which the target substance is absent. ### Claim 2 (depends on 1) The screening method of claim 1 wherein the Semaphorin 6C is a human, mouse or rat Semaphorin 6C. ### Claim 3 (depends on 1) The screening method for agonists or antagonists of Semaphorin 6C according to claim 1. ### Claim 4 (depends on 1) The screening method for agonists or antagonists for Semaphorin 6C according to claim 1, wherein the protein having the extracellular domain of Semaphorin 6C is a protein bound to a marker protein and/or a peptide tag. ### Claim 5 (depends on 4) The screening method for agonists or antagonists for Semaphorin 6C according to claim 4, wherein the marker protein is alkaline phosphatase or an immunoglobulin Fc domain. ### Claim 6 (depends on 1) The screening method for agonists or antagonists for Semaphorin 6C according to claim 1, wherein a cell membrane or a cell expressing a protein with the extracellular domain of Plexin-A1 is used. ### Claim 7 (depends on 6) The screening method for agonists or antagonists for Semaphorin 6C according to claim 6, wherein the cell membrane or cell expressing protein having the extracellular domain of Plexin-A1 is, or is obtained from, a stable transformant. ### Claim 8 (depends on 1) The screening method for agonists or antagonists for Semaphorin 6C according to claim 1, wherein the protein having the extracellular domain of Plexin-A1 is a recombinant protein. ### Claim 9 (depends on 1) The screening method for agonists or antagonists for Semaphorin 6C according to claim 1, wherein the protein having the extracellular domain of Plexin-A1 is Plexin-A1. ### Claim 10 A screening method for agonists or antagonists of Semaphorin 6C, comprising the steps of 1) contacting Plexin-A1 consisting of SEQ ID NO:5 or SEQ D NO:6 with a target substance in an in vitro assay system, and 2) measuring Semaphorin 6C activity in said assay system by detecting the presence or absence of a signal arising from an interaction between a protein having an extracellular domain of Semaphorin 6C comprising the 1st to 599th amino acid residue in the sequence of Semaphorin 6C consisting of SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10 and Plexin-A1, wherein the signal is i) a growth cone collapse activity, or ii) a contractile activity of a Plexin-A1 expressing cell, wherein a decrease in said Semaphorin 6C activity indicates that said target substance is an antagonist and an increase in said Semaphorin 6C activity indicates that said target substance is an agonist. ### Claim 11 (depends on 10) The screening method of claim 10 wherein the Semaphorin 6C is a human, mouse or rat Semaphorin C. ### Claim 12 (depends on 10) The screening method for agonists or antagonists of Semaphorin 6C according to claim 10, wherein the protein is recombinant.