# US 11,306,150 B2 · Method of identifying a P-selectin glycoprotein ligand-1 (PSGL-1) antagonist > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,306,150 B2 - Document ID: US11306150B2 - Status: Granted - Kind: B2 - Application number: 16/476,814 - Filed: 2018-01-10 - Published: 2019-10-31 - Granted: 2022-04-19 - Earliest claimed: 2017-01-11 - Assignees: Bristol-Myers Squibb Company; Five Prime Therapeutics, Inc. - Applicants: Bristol-Myers Squibb Company; Five Prime Therapeutics, Inc. - Inventors: Robert J. Johnston; Andrew Rankin; Arathi Krishnakumar; Paul O. Sheppard; Arvind Rajpal - Web page: https://patentgrep.com/patent/US11306150B2 ## CPC - C07K 16/2896: {against molecules with a "CD"-designation, not provided for elsewhere} - A61P 35/00: Antineoplastic agents - A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca - C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered - C07K 2317/54: F(ab')2 - C07K 2317/55: Fab or Fab' - C07K 2317/56: variable (Fv) region, i.e. VH and/or VL - C07K 2317/76: Antagonist effect on antigen, e.g. neutralization or inhibition of binding - C07K 2319/30: Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto - C07K 2319/32: fusions with soluble part of a cell surface receptor, "decoy receptors" - 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) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) ## Abstract Methods of identifying and using PSGL-1 antagonists are provided. Such methods include, but are not limited to, methods of treating cancer. PSGL-1 antagonists include, but are not limited to, antibodies that bind PSGL-1 and antibodies that bind VISTA, wherein the antibodies inhibit PSGL-1 binding to VISTA, e.g., at acidic pH (e.g., pH 6.0), as well as PSGL-1 and VISTA extracellular domain polypeptides. ## Claims ### Claim 1 A method of identifying a PSGL-1 antagonist, comprising: a) forming a composition comprising a candidate molecule, a VISTA molecule, and a PSGL-1 molecule, wherein the VISTA molecule comprises VISTA, a VISTA ECD, or a VISTA ECD fusion molecule, and the PSGL-1 molecule comprises PSGL-1, PSGL-1 ECD, or a PSGL-1 ECD fusion molecule; and b) detecting binding of the VISTA molecule to the PSGL-1 molecule; wherein the composition has a pH in the range of pH 5.5 to pH 6.5, and wherein a reduction in the binding of the VISTA molecule to the PSGL-1 molecule in the presence of the candidate molecule as compared to the binding of the VISTA molecule to the PSGL-1 molecule in the absence of the candidate molecule indicates that the candidate molecule is a PSGL-1 antagonist wherein the PSGL-1 ECD comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 3, 4, 16, 17, or 18, and the VISTA ECD comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of amino acids 1-202 of SEQ ID NO: 10. ### Claim 2 (depends on 1) The method of claim 1, wherein the candidate molecule is selected from a PSGL-1 antibody and a VISTA antibody. ### Claim 3 (depends on 1) The method of claim 1, wherein binding of the VISTA molecule to the PSGL-1 molecule is reduced by at least 30% in the presence of the candidate molecule. ### Claim 4 (depends on 1) The method of claim 1, wherein binding of the VISTA molecule to the PSGL-1 molecule is detected by a method selected from surface plasmon resonance, ELISA, amplified luminescent proximity homogeneous assay, and flow cytometry. ### Claim 5 (depends on 1) The method of claim 1, wherein the VISTA molecule is expressed on the surface of a cell. ### Claim 6 (depends on 1) The method of claim 1, wherein the PSGL-1 molecule is expressed on the surface of a cell. ### Claim 7 (depends on 1) The method of claim 1, wherein the candidate molecule is selected from a small molecule and a small peptide. ### Claim 8 (depends on 1) The method of claim 1, wherein the composition has a pH of 6.0 to pH 6.5. ### Claim 9 (depends on 8) The method of claim 8, wherein the composition has a pH of 6.0. ### Claim 10 (depends on 1) The method of claim 1, wherein the PSGL-1 molecule is a PSGL-1 ECD or PSGL-1 ECD fusion protein, wherein the PSGL-1 ECD comprises the amino acid sequence of SEQ ID NO: 3, 4, 16, 17, or 18. ### Claim 11 (depends on 8) The method of claim 8, wherein the PSGL-1 molecule is a PSGL-1 ECD or PSGL-1 ECD fusion protein, wherein the PSGL-1 ECD comprises the amino acid sequence of SEQ ID NO: 3, 4, 16, 17, or 18. ### Claim 12 (depends on 1) The method of claim 1, wherein the VISTA molecule is a VISTA ECD or VISTA ECD fusion protein, wherein the VISTA ECD comprises the amino acid sequence of amino acids 1-202 of SEQ ID NO: 10 or comprises the amino acid sequence of SEQ ID NO: 9. ### Claim 13 (depends on 8) The method of claim 8, wherein the VISTA molecule is a VISTA ECD or VISTA ECD fusion protein, wherein the VISTA ECD comprises the amino acid sequence of amino acids 1-202 of SEQ ID NO: 10 or comprises the amino acid sequence of SEQ ID NO: 9. ### Claim 14 (depends on 1) The method of claim 1, wherein the PSGL-1 molecule comprises the amino acid sequence of SEQ ID NO: 1, 2, 14, or 15. ### Claim 15 (depends on 1) The method of claim 1, wherein the VISTA molecule comprises the amino acid sequence of SEQ ID NO: 5 or 6. ### Claim 16 (depends on 8) The method of claim 8, wherein the PSGL-1 molecule comprises the amino acid sequence of SEQ ID NO: 1, 2, 14, or 15. ### Claim 17 (depends on 8) The method of claim 8, wherein the VISTA molecule comprises the amino acid sequence of SEQ ID NO: 5 or 6.