# US 10,501,553 B2 · Methods and compositions for cancer treating conditions relating to over expressions of EphA2 > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,501,553 B2 - Document ID: US10501553B2 - Status: Granted - Kind: B2 - Application number: 15/528,073 - Filed: 2015-11-18 - Published: 2017-12-07 - Granted: 2019-12-10 - Earliest claimed: 2014-11-19 - Applicants: Memorial Sloan-Kettering Cancer Center; University of Turku - Inventors: Dimitar B Nikolov; Juha Himanen; Urpo Lamminmaki; Hanna Sanmark - Web page: https://patentgrep.com/patent/US10501553B2 ## CPC - C07K 16/2887: {against CD20} - A61K 39/395: Antibodies (agglutinins A61K38/36 {; as drug carriers A61K47/50}); Immunoglobulins; Immune serum, e.g. antilymphocytic serum - A61K 39/3955: {against proteinaceous materials, e.g. enzymes, hormones, lymphokines} - C07K 16/2866: {against receptors for cytokines, lymphokines, interferons} - C07K 16/2896: {against molecules with a "CD"-designation, not provided for elsewhere} - C07K 16/30: from tumour cells - C07K 16/468: {Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies} - A61K 2039/505: {comprising antibodies} - A61K 2039/545: {characterised by the dose, timing or administration schedule} - C07K 2317/31: multispecific - C07K 2317/622: Single chain antibody (scFv) - C07K 2317/73: Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation - C07K 2317/76: Antagonist effect on antigen, e.g. neutralization or inhibition of binding - C07K 2319/00: Fusion polypeptide - 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) - 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 An anti-EphA2 scFv single-chain antibody binds with high specificity for EphA2 and blocks ephrin binding to Eph-A2. The antibody may be linked with other antigen-targeting domains, such as an anti-CD20 domain, or conjugated with toxins, or used in combination therapy to for treatment of conditions related to overexpression of EphA2. The antibody may also be expressed in vivo by an expression vector that is designed to facilitate such treatment. ## Claims ### Claim 1 A protein comprising an anti-EphA2 scFv domain comprising the amino acid sequence of SEQ ID NO: 1, 5, 6 or 7. ### Claim 2 (depends on 1) A protein according to claim 1, further comprising an anti-CD20 scFV domain and a peptide linker, wherein the peptide linker connects the anti-EphA2 scFv domain to the anti-CD20 scFV domain. ### Claim 3 (depends on 2) A protein according to claim 2, wherein the anti-EphA2 scFv domain comprises the amino acid sequence of SEQ ID NO: 1. ### Claim 4 (depends on 2) A protein according to claim 2, further comprising an N-terminal signal peptide sequence for secretion of the protein. ### Claim 5 (depends on 2) A polynucleotide comprising a nucleotide sequence encoding the protein of claim 2. ### Claim 6 (depends on 5) An expression vector comprising the polynucleotide of claim 5 operably linked to one or more regulatory sequences sufficient for expressing the protein in a cell. ### Claim 7 (depends on 6) A cultured cell comprising the expression vector of claim 6. ### Claim 8 (depends on 7) A method for producing a protein comprising an anti-EphA2 scFv domain, said method comprising culturing the cell of claim 7 to produce the protein. ### Claim 9 (depends on 2) A method of inhibiting the proliferation of lymphoma or leukemia cells that express EphA2, the method comprising contacting the lymphoma or leukemia cells in vitro with an effective amount of a protein according to claim 2. ### Claim 10 (depends on 1) A protein according to claim 1, further comprising an anti-CD20 scFV domain comprising the amino acid sequence of SEQ ID NO:3 and a peptide linker, wherein the peptide linker connects the anti-EphA2 scFv domain to the anti-CD20 scFV domain. ### Claim 11 (depends on 10) A protein according to claim 10, wherein the anti-EphA2 scFv domain comprises the amino acid sequence of SEQ ID NO: 1. ### Claim 12 (depends on 10) A protein according to claim 10, comprising the amino acid sequence of SEQ ID NO:4 or SEQ ID NO:9. ### Claim 13 (depends on 10) A protein according to claim 10, further comprising an N-terminal signal peptide sequence for secretion of the protein. ### Claim 14 (depends on 10) A polynucleotide comprising a nucleotide sequence encoding the protein of claim 10. ### Claim 15 (depends on 14) An expression vector comprising the polynucleotide of claim 14 operably linked to one or more regulatory sequences sufficient for expressing the protein in a cell. ### Claim 16 (depends on 13) A cultured cell comprising the expression vector of claim 13. ### Claim 17 (depends on 16) A method for producing a protein comprising an anti-EphA2 scFv domain, said method comprising culturing the cell of claim 16 to produce the protein. ### Claim 18 (depends on 14) A polynucleotide according to claim 14, wherein the polynucleotide comprises SEQ ID NO:2 or SEQ ID NO:8. ### Claim 19 (depends on 18) An expression vector comprising the polynucleotide of claim 18 operably linked to one or more regulatory sequences sufficient for expressing the protein in a cell. ### Claim 20 (depends on 19) A cultured cell comprising the expression vector of claim 19. ### Claim 21 (depends on 20) A method for producing a protein comprising an anti-EphA2 scFv domain, said method comprising culturing the cell of claim 20 to produce the protein. ### Claim 22 (depends on 10) A method of inhibiting the proliferation of lymphoma or leukemia cells that express EphA2, the method comprising contacting the lymphoma or leukemia cells in vitro with an effective amount of a protein according to claim 10. ### Claim 23 (depends on 1) A protein according to claim 1, wherein the anti-EphA2 scFv domain comprises the amino acid sequence of SEQ ID NO: 1. ### Claim 24 (depends on 1) A protein according to claim 1, further comprising an N-terminal signal peptide sequence for secretion of the protein. ### Claim 25 (depends on 1) A polynucleotide comprising a nucleotide sequence encoding the protein of claim 1. ### Claim 26 (depends on 25) An expression vector comprising the polynucleotide of claim 25 operably linked to one or more regulatory sequences sufficient for expressing the protein in a cell. ### Claim 27 (depends on 1) A method of inhibiting the proliferation of lymphoma or leukemia cells that express EphA2, the method comprising contacting the lymphoma or leukemia cells in vitro with an effective amount of a protein according to claim 1.