# US 2008/0187532 A1 · Compositions and methods for diagnosing and treating cancer > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2008/0187532 A1 - Document ID: US20080187532A1 - Status: Published application - Kind: A1 - Application number: 11/905,392 - Filed: 2007-09-28 - Published: 2008-08-07 - Earliest claimed: 2006-09-29 - Assignees: OncoMed Pharmaceuticals, Inc. - Inventors: Austin Gurney; Fumiko Axelrod; Tim Hoey; Sanjeev Satyal - Web page: https://patentgrep.com/patent/US20080187532A1 ## CPC - A61K 39/39558: {against tumor tissues, cells, antigens} - A61K 31/4745: condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines (yohimbine derivatives, vinblastine A61K31/475; ergoline derivatives A61K31/48) - A61K 31/513: having oxo groups directly attached to the heterocyclic ring, e.g. cytosine - A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca - A61N 5/10: X-ray therapy; Gamma-ray therapy; Particle-irradiation therapy (A61N5/01 takes precedence) - A61P 35/00: Antineoplastic agents - A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 - C07K 16/30: from tumour cells - C07K 16/3015: {Breast} - C07K 16/3023: {Lung} - C07K 16/303: {Liver or Pancreas} - C07K 16/3046: {Stomach, Intestines} - C07K 16/3069: {Reproductive system, e.g. ovaria, uterus, testes, prostate} - A61K 2039/505: {comprising antibodies} - A61K 2039/507: {Comprising a combination of two or more separate antibodies} - C07K 2317/21: from primates, e.g. man - C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered - C07K 2317/34: Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues - C07K 2317/56: variable (Fv) region, i.e. VH and/or VL - C07K 2317/565: Complementarity determining region [CDR] - C07K 2317/73: Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation - C07K 2317/732: Antibody-dependent cellular cytotoxicity [ADCC] - C07K 2317/734: Complement-dependent cytotoxicity [CDC] - C07K 2317/76: Antagonist effect on antigen, e.g. neutralization or inhibition of binding - 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) - A61N: ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY (measurement of bioelectric currents A61B; surgical instruments, devices or methods for transferring non-mechanical forms of energy to or from the body A61B18/00; anaesthetic apparatus in general A61M; incandescent lamps H01K; infrared radiators for heating H05B) - 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 An isolated antibody that specifically binds to an extracellular domain of human DLL4 and affects growth of a tumor comprising cancer stem cells is described. Also described is a method of treating cancer comprising administering a therapeutically effective amount of an anti-DLL4 antibody. ## Claims ### Claim 1 An isolated antibody that specifically binds to a human DLL4 epitope comprising amino acids within human DLL4 N-terminal region (SEQ ID NO: 27), wherein the antibody affects growth of a tumor comprising cancer stem cells. ### Claim 2 (canceled) ### Claim 3 (depends on 1) The isolated antibody of claim 1, wherein the human DLL4 epitope further comprises amino acids within human DLL4 DSL (SEQ ID NO: 26). ### Claim 4 (depends on 1) The isolated antibody of claim 1, wherein the human DLL4 epitope comprises amino acids 66-73 (QAVVSPGP) of the human DLL4 N-terminal region. ### Claim 5 (canceled) ### Claim 6 (depends on 1) The isolated antibody of claim 1, wherein the human DLL4 epitope comprises amino acids 139-146 (LISKIAIQ) of the human DLL4 N-terminal region. ### Claim 7 (canceled) ### Claim 8 (depends on 1) The isolated antibody of claim 1, wherein the human DLL4 epitope comprises amino acids 66-73 (QAVVSPGP) and 139-146 (LISKIAIQ) of the human DLL4 N-terminal region. ### Claim 9 (canceled) ### Claim 10 (depends on 1) The isolated antibody of claim 1, wherein the affect is a reduction in tumor volume. ### Claim at position 11 11-12. (canceled) ### Claim 13 (depends on 1) The antibody of claim 1, wherein the antibody is a humanized antibody or human antibody. ### Claim at position 13 14-18. (canceled) ### Claim 19 (depends on 1) The isolated antibody of claim 1 comprising: i. a heavy chain variable region comprising CDR amino acid sequences CDR1 (SEQ ID NO: 1); CDR2 (SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4); and CDR3 (SEQ ID NO: 5); and ii. a light chain variable region comprising CDR amino acid sequences CDR1 (SEQ ID NO: 9); CDR2 (SEQ ID NO: 10); and CDR3 (SEQ ID NO: 11). ### Claim 20 (depends on 19) The isolated antibody of claim 19 produced by hybridoma deposited with ATCC on Sep. 28, 2007 and having ATCC deposit no. PTA-8670. ### Claim 21 An antibody that competes for specific binding to human DLL4 with an antibody produced by hybridoma deposited with ATCC on Sep. 28, 2007 and having ATCC deposit no. PTA-8670. ### Claim 22 A humanized antibody that specifically binds to a human DLL4 epitope comprising amino acids within human DLL4 N-terminal region (SEQ ID NO: 27), wherein the antibody comprises: (i) a heavy chain variable region comprising a nonhuman antigen determination region and a human heavy chain variable framework region, and (ii) a light chain variable region comprising a nonhuman antigen determination region and a human light chain variable framework region. ### Claim 23 (depends on 22) The humanized antibody of claim 22, wherein the human DLL4 epitope is formed by a combination of the human DLL4 N-terminal region (SEQ ID NO: 27) and human DLL4 DSL (SEQ ID NO: 26). ### Claim 24 (canceled) ### Claim 25 (depends on 23) The humanized antibody of claim 23, wherein at least one residue in the human heavy chain variable framework region is substituted. ### Claim at position 21 26-27. (canceled) ### Claim 28 (depends on 25) The humanized antibody of claim 25, wherein the heavy chain nonhuman antigen determination region comprises CDR amino acid sequences CDR1 (SEQ ID NO: 1); CDR2 (SEQ ID NO: 2, SEQ ID NO: 3 or SEQ ID NO: 4); and CDR3 (SEQ ID NO: 5). ### Claim 29 (depends on 23) The humanized antibody of claim 23, wherein at least one residue in the human light chain variable framework region is substituted. ### Claim at position 24 30-31. (canceled) ### Claim 32 (depends on 29) The humanized antibody of claim 29, wherein the light chain nonhuman antigen determination region comprises CDR1 (SEQ ID NO: 9); CDR2 (SEQ ID NO: 10); and CDR3 (SEQ ID NO: 11). ### Claim 33 (depends on 23) The humanized antibody of claim 23, wherein at least one residue in the human heavy chain variable framework region is substituted and at least one residue in the human light chain variable framework region is substituted. ### Claim 34 (depends on 33) The humanized antibody of claim 33, wherein five or six residues in the human heavy chain variable framework region are substituted and two residues in the human light chain variable framework region are substituted. ### Claim 35 (depends on 34) The humanized antibody of claim 34, wherein the heavy chain nonhuman antigen determination region comprises CDR amino acid sequences CDR1 (SEQ ID NO: 1); CDR2 (SEQ ID NO: 2, SEQ ID NO: 3 or SEQ ID NO: 4); and CDR3 (SEQ ID NO: 5); and the chain light nonhuman antigen determination region comprises CDR1 (SEQ ID NO: 9); CDR2 (SEQ ID NO: 10); and CDR3 (SEQ ID NO: 11). ### Claim 36 An isolated antibody that specifically binds to a human DLL4 epitope comprising amino acids within human DLL4 N-terminal region (SEQ ID NO: 27), wherein the antibody inhibits DLL4 binding to a Notch receptor. ### Claim 37 A pharmaceutical composition comprising: i. an isolated antibody that specifically binds to a human DLL4 epitope comprising amino acids within human DLL4 N-terminal region (SEQ ID NO: 27), wherein the antibody affects growth of a tumor comprising cancer stem cells; and ii. a pharmaceutically acceptable carrier. ### Claim 38 A pharmaceutical composition comprising: i. an isolated antibody that competes for specific binding to human DLL4 with an antibody produced by hybridoma deposited with ATCC on Sep. 28, 2007 and having ATCC deposit no. PTA-8670. ii. a pharmaceutically acceptable carrier. ### Claim 39 An isolated polynucleotide vector encoding an antibody that specifically binds to a human DLL4 epitope comprising amino acids within human DLL4 N-terminal region (SEQ ID NO: 27), wherein the antibody affects growth of a tumor comprising cancer stem cells. ### Claim 40 (depends on 39) The isolated polynucleotide vector of claim 39, wherein the antibody comprises: i. a heavy chain variable region comprising CDR amino acid sequences CDR1 (SEQ ID NO: 1); CDR2 (SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4); and CDR3 (SEQ ID NO 5); and ii. a light chain variable region comprising CDR amino acid sequences CDR1 (SEQ ID NO: 9); CDR2 (SEQ ID NO: 10); and CDR3 (SEQ ID NO: 11). ### Claim 41 (depends on 40) The isolated polynucleotide vector of claim 40, wherein: i. the heavy chain variable region polynucleotide sequences are CDR1 (SEQ ID NO: 17), CDR2 (SEQ ID NO: 18, SEQ ID NO: 19, or SEQ ID NO: 20); and CDR3 (SEQ ID NO: 21); and ii. the light chain variable region polynucleotide sequences are CDR1 (SEQ ID NO: 22); CDR2 (SEQ ID NO: 23); and CDR3 (SEQ ID NO: 24). ### Claim 42 An isolated polynucleotide vector encoding a heavy chain variable region of an antibody that specifically binds to a human DLL4 epitope comprising amino acids within human DLL4 N-terminal region (SEQ ID NO: 27), wherein the antibody affects growth of a tumor comprising cancer stem cells. ### Claim 43 (depends on 42) The isolated polynucleotide vector of claim 42, wherein the antibody comprises a heavy chain variable region comprising CDR amino acid sequences CDR1 (SEQ ID NO: 1); CDR2 (SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4); and CDR3 (SEQ ID NO: 5). ### Claim 44 (depends on 43) The isolated polynucleotide vector of claim 43, comprising heavy chain variable region polynucleotide sequences CDR1 (SEQ ID NO: 17), CDR2 (SEQ ID NO: 18, SEQ ID NO: 19, or SEQ ID NO: 20); and CDR3 (SEQ ID NO: 21). ### Claim 45 An isolated polynucleotide vector encoding a light chain variable region of an antibody that specifically binds to a human DLL4 epitope comprising amino acids within human DLL4 N-terminal region (SEQ ID NO: 27), wherein the antibody affects growth of a tumor comprising cancer stem cells. ### Claim 46 (depends on 45) The isolated polynucleotide vector of claim 45, wherein the antibody comprises a light chain variable region comprising CDR amino acid sequences CDR1 (SEQ ID NO: 9); CDR2 (SEQ ID NO: 10); and CDR3 (SEQ ID NO: 11). ### Claim 47 (depends on 46) The isolated polynucleotide vector of claim 46, comprising light chain variable region polynucleotide sequences CDR1 (SEQ ID NO: 22), CDR2 (SEQ ID NO: 23); and CDR3 (SEQ ID NO: 24). ### Claim 48 A host cell comprising one or more polynucleotide vectors encoding an antibody, wherein said antibody specifically binds to a human DLL4 epitope comprising amino acids within human DLL4 N-terminal region (SEQ ID NO: 27), wherein the antibody affects growth of a tumor comprising cancer stem cells. ### Claim 49 A method of treating cancer comprising administering to a patient a therapeutically effective amount of an antibody that specifically binds to a human DLL4 epitope comprising amino acids within human DLL4 N-terminal region (SEQ ID NO: 27). ### Claim 50 A method of treating cancer comprising administering to a patient a therapeutically effective amount of a humanized antibody that specifically binds to a human DLL4 epitope comprising amino acids within human DLL4 N-terminal region (SEQ ID NO: 27), wherein the antibody comprises: (i) a heavy chain variable region comprising a nonhuman antigen determination region and a human heavy chain variable framework region, and (ii) a light chain variable region comprising a nonhuman antigen determination region and a human light chain variable framework region. ### Claim 51 A method of treating cancer comprising administering to a patient a therapeutically effective amount of an antibody that specifically to a human DLL4 epitope comprising amino acids within human DLL4 N-terminal region (SEQ ID NO: 27) in combination with a second therapeutic agent. ### Claim 52 (canceled) ### Claim 53 (depends on 51) The method of claim 51, wherein the second therapeutic agent is a chemotherapeutic agent or radiation therapy. ### Claim 54 (canceled) ### Claim 55 (depends on 51) The method of claim 51, wherein the second therapeutic agent is a second therapeutic antibody. ### Claim at position 49 56-57. (canceled) ### Claim 58 A method of diagnosing cancer comprising contacting a patient sample with an antibody that specifically binds to a human DLL4 epitope comprising amino acids within human DLL4 N-terminal region (SEQ ID NO: 27), and diagnosing cancer based on the localization of DLL4 expression in the patient sample. ### Claim 59 A method of manufacturing an antibody that specifically binds to a human DLL4 epitope comprising amino acids within human DLL4 N-terminal region (SEQ ID NO: 27), the method comprising: i. transforming host cells with a polynucleotide expression vector or vectors encoding the antibody; ii. culturing the host cells under conditions permitting expression of the antibody; and iii. purifying the antibody from the culture medium. ### Claim 60 A transformed cell line expressing an antibody that specifically binds to a human DLL4 epitope comprising amino acids within human DLL4 N-terminal region (SEQ ID NO: 27). ### Claim 61 A hybridoma that secretes antibody 21M18 deposited with ATCC on Sep. 28, 2007 and having ATCC deposit no. PTA-8670. ### Claim 62 A method of producing antibody 21M18 by culturing a hybridoma deposited with ATCC on Sep. 28, 2007 and having ATCC deposit no. PTA-8670. ### Claim 63 A monoclonal antibody produced by hybridoma deposited with ATCC on Sep. 28, 2007 and having ATCC deposit no. PTA-8670. ### Claim 64 A kit comprising a container and a composition contained therein, wherein the composition comprises an antibody that binds to a human DLL4 epitope comprising amino acids within human DLL4 N-terminal region (SEQ ID NO: 27), and further comprises a package insert indicating that the composition can be used to treat cancer.