Therapeutic DLL4 binding proteins

• US 9,469,689 B2
• Filed: 07/14/2015
• Issued: 10/18/2016
• Est. Priority Date: 03/02/2010
• Status: Active Grant

• Current Assignee: Abbvie Incorporated
• Original Assignee: Abbvie Incorporated
• Inventors: Morgan-Lappe, Susan E., Chen, Ming-Jiu, Hsieh, Chung-Ming, Gu, Jijie, Li, Yingchun
• Primary Examiner(s): Goddard, Laura B
• Assistant Examiner(s): Natarajan, Meera
• Application Number:
  US14/798,849
• Publication Number:
  US 20160031986A1
• Time in Patent Office: 462 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/73 Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Abstract

DLL4 binding proteins are described herein, including antibodies, CDR-grafted antibodies, humanized antibodies, and DLL4 binding fragments thereof, proteins that bind DLL4 with high affinity, and DLL4 binding proteins that neutralize DLL4 and/or VEGF activity. The DLL4 binding proteins are useful for treating or preventing cancers and tumors and especially for treating or preventing tumor angiogenesis.

Claims

1. A binding protein comprising an antigen-binding domain capable of binding DLL4, wherein the antigen-binding domain comprises a set of six complementarity determining region (CDR) sequences:

CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3, wherein;

(a) CDR-H1 is selected from the group consisting of;

residues 31-35 of SEQ ID NO;

157 (CDR-H1 38H12);

residues 31-35 of SEQ ID NO;

161 (CDR-H1 37D10);

residues 31-35 of SEQ ID NO;

163 (CDR-H1 32C7);

residues 31-35 of SEQ ID NO;

165 (CDR-H1 14G1);

residues 31-35 of SEQ ID NO;

167 (CDR-H1 14A11);

residues 31-35 of SEQ ID NO;

169 (CDR-H1 15D6);

residues 31-35 of SEQ ID NO;

179 (CDR-H1 VH.1 38H12);

residues 31-35 of SEQ ID NO;

180 (CDR-H1 VH.1a 38H12);

residues 31-35 of SEQ ID NO;

181 (CDR-H1 VH.1b 38H12); and

residues 31-35 of SEQ ID NO;

182 (CDR-H1 VH.2a 38H12);

(b) CDR-H2 is selected from the group consisting of;

residues 50-66 of SEQ ID NO;

157 (CDR-H2 38H12);

residues 50-68 of SEQ ID NO;

161 (CDR-H2 37D10);

residues 50-66 of SEQ ID NO;

163 (CDR-H2 32C7);

residues 50-66 of SEQ ID NO;

165 (CDR-H2 14G1);

residues 50-66 of SEQ ID NO;

167 (CDR-H2 14A11);

residues 50-66 of SEQ ID NO;

169 (CDR-H2 15D6);

residues 50-66 of SEQ ID NO;

179 (CDR-H2 VH.1 38H12);

residues 50-66 of SEQ ID NO;

180 (CDR-H2 VH.1a 38H12);

residues 50-66 of SEQ ID NO;

181 (CDR-H2 VH.1 b 38H12); and

residues 50-66 of SEQ ID NO;

182 (CDR-H2 VH.2a 38H12);

(c) CDR-H3 is selected from the group consisting of;

residues 99-107 of SEQ ID NO;

157 (CDR-H3 38H12);

residues 101-111 of SEQ ID NO;

161 (CDR-H3 37D10);

residues 99-105 of SEQ ID NO;

163 (CDR-H3 32C7);

residues 99-105 of SEQ ID NO;

165 (CDR-H3 14G1);

residues 99-110 of SEQ ID NO;

167 (CDR-H3 14A11);

residues 99-110 of SEQ ID NO;

169 (CDR-H3 15D6);

residues 99-107 of SEQ ID NO;

179 (CDR-H3 VH.1 38H12);

residues 99-107 of SEQ ID NO;

180 (CDR-H3 VH.1a 38H12);

residues 99-107 of SEQ ID NO;

181 (CDR-H3 VH.1 b 38H12); and

residues 99-107 of SEQ ID NO;

182 (CDR-H3 VH.2a 38H12);

(d) CDR-L1 is selected from the group consisting of;

residues 24-34 of SEQ ID NO;

158 (CDR-L1 38H12);

residues 24-34 of SEQ ID NO;

162 (CDR-L1 37D10);

residues 24-34 of SEQ ID NO;

164 (CDR-L1 32C7);

residues 24-34 of SEQ ID NO;

166 (CDR-L1 14G1);

residues 23-37 of SEQ ID NO;

168 (CDR-L1 14A11);

residues 23-37 of SEQ ID NO;

170 (CDR-L1 15D6);

residues 24-34 of SEQ ID NO;

183 (CDR-L1 VL.1 38H12);

residues 24-34 of SEQ ID NO;

184 (CDR-L1 VL.1a 38H12);

residues 24-34 of SEQ ID NO;

185 (CDR-L1 VL.1b 38H12); and

residues 24-34 of SEQ ID NO;

186 (CDR-L1 VL.2a 38H12);

(e) CDR-L2 is selected from group consisting of;

residues 50-56 of SEQ ID NO;

158 (CDR-L2 38H12);

residues 50-56 of SEQ ID NO;

162 (CDR-L2 37D10);

residues 50-56 of SEQ ID NO;

164 (CDR-L2 32C7);

residues 50-56 of SEQ ID NO;

166 (CDR-L2 14G1);

residues 53-59 of SEQ ID NO;

168 (CDR-L2 14A11);

residues 53-59 of SEQ ID NO;

170 (CDR-L2 15D6);

residues 50-56 of SEQ ID NO;

183 (CDR-L2 VL.1 38H12);

residues 50-56 of SEQ ID NO;

184 (CDR-L2 VL.1a 38H12);

residues 50-56 of SEQ ID NO;

185 (CDR-L2 VL.1b 38H12); and

residues 50-56 of SEQ ID NO;

186 (CDR-L2 VL.2a 38H12);

and

(f) CDR-L3 is selected from the group consisting of;

residues 89-97 of SEQ ID NO;

158 (CDR-L3 38H12);

residues 89-97 of SEQ ID NO;

162 (CDR-L3 37D10);

residues 89-98 of SEQ ID NO;

164 (CDR-L3 32C7);

residues 89-97 of SEQ ID NO;

166 (CDR-L3 14G1);

residues 92-100 of SEQ ID NO;

168 (CDR-L3 14A11);

residues 92-100 of SEQ ID NO;

170 (CDR-L3 15D6);

residues 89-97 of SEQ ID NO;

183 (CDR-L3 VL.1 38H12);

residues 89-97 of SEQ ID NO;

184 (CDR-L3 VL.1a 38H12);

residues 89-97 of SEQ ID NO;

185 (CDR-L3 VL.1b 38H12); and

residues 89-97 of SEQ ID NO;

186 (CDR-L3 VL.2a 38H12).

2. The binding protein according to claim 1, wherein the binding protein comprises a CDR set of three CDRs selected from the group consisting of:

VH 38H12 CDR Set, comprising;

CDR-H1;

residues 31-35 of SEQ ID NO;

157

CDR-H2;

residues 50-66 of SEQ ID NO;

157 and

CDR-H3;

residues 99-107 of SEQ ID NO;

157

VL 38H12 CDR Set, comprising;

CDR-L1;

residues 24-34 of SEQ ID NO;

158

CDR-L2;

residues 50-56 of SEQ ID NO;

158 and

CDR-L3;

residues 89-97 of SEQ ID NO;

158

VH 37D10 CDR Set, comprising;

CDR-H1;

residues 31-35 of SEQ ID NO;

161

CDR-H2;

residues 50-68 of SEQ ID NO;

161 and

CDR-H3;

residues 101-111 of SEQ ID NO;

161

VL 37D10 CDR Set, comprising;

CDR-L1;

residues 24-34 of SEQ ID NO;

162

CDR-L2;

residues 50-56 of SEQ ID NO;

162 and

CDR-L3;

residues 89-97 of SEQ ID NO;

162

VH 32C7 CDR Set, comprising;

CDR-H1;

residues 31-35 of SEQ ID NO;

163

CDR-H2;

residues 50-66 of SEQ ID NO;

163 and

CDR-H3;

residues 99-105 of SEQ ID NO;

163

VL 32C7 CDR Set, comprising;

CDR-L1;

residues 24-34 of SEQ ID NO;

164

CDR-L2;

residues 50-56 of SEQ ID NO;

164 and

CDR-L3;

residues 89-98 of SEQ ID NO;

164

VH 14G1 CDR Set, comprising;

CDR-H1;

residues 31-35 of SEQ ID NO;

165

CDR-H2;

residues 50-66 of SEQ ID NO;

165 and

CDR-H3;

residues 99-105 of SEQ ID NO;

165

VL 14G1 CDR Set, comprising;

CDR-L1;

residues 24-34 of SEQ ID NO;

166

CDR-L2;

residues 50-56 of SEQ ID NO;

166 and

CDR-L3;

residues 89-97 of SEQ ID NO;

166

VH 14A11 CDR Set, comprising;

CDR-H1;

residues 31-35 of SEQ ID NO;

167

CDR-H2;

residues 50-66 of SEQ ID NO;

167 and

CDR-H3;

residues 99-110 of SEQ ID NO;

167

VL 14A11 CDR Set, comprising;

CDR-L1;

residues 23-37 of SEQ ID NO;

168

CDR-L2;

residues 53-59 of SEQ ID NO;

168 and

CDR-L3;

residues 92-100 of SEQ ID NO;

168

VH 15D6 CDR Set, comprising;

CDR-H1;

residues 31-35 of SEQ ID NO;

169

CDR-H2;

residues 50-66 of SEQ ID NO;

169 and

CDR-H3;

residues 99-110 of SEQ ID NO;

169

VL 15D6 CDR Set, comprising;

CDR-L1;

residues 23-37 of SEQ ID NO;

170

CDR-L2;

residues 53-59 of SEQ ID NO;

170 and

CDR-L3;

residues 92-100 of SEQ ID NO;

170

VH.1 38H12 CDR Set, comprising;

CDR-H1;

residues 31-35 of SEQ ID NO;

179

CDR-H2;

residues 50-66 of SEQ ID NO;

179 and

CDR-H3;

residues 99-107 of SEQ ID NO;

179

VH.1a 38H12 CDR Set, comprising;

CDR-H1;

residues 31-35 of SEQ ID NO;

180

CDR-H2;

residues 50-66 of SEQ ID NO;

180 and

CDR-H3;

residues 99-107 of SEQ ID NO;

180

VH.1b 38H12 CDR Set, comprising;

CDR-H1;

residues 31-35 of SEQ ID NO;

181

CDR-H2;

residues 50-66 of SEQ ID NO;

181 and

CDR-H3;

residues 99-107 of SEQ ID NO;

181

VH.2a 38H12 CDR Set, comprising;

CDR-H1;

residues 31-35 of SEQ ID NO;

182

CDR-H2;

residues 50-66 of SEQ ID NO;

182 and

CDR-H3;

residues 99-107 of SEQ ID NO;

182

VL.1 38H12 CDR Set, comprising;

CDR-L1;

residues 24-34 of SEQ ID NO;

183

CDR-L2;

residues 50-56 of SEQ ID NO;

183 and

CDR-L3;

residues 89-97 of SEQ ID NO;

183

VL.1a 38H12 CDR Set, comprising;

CDR-L1;

residues 24-34 of SEQ ID NO;

184

CDR-L2;

residues 50-56 of SEQ ID NO;

184 and

CDR-L3;

residues 89-97 of SEQ ID NO;

184

VL.1b 38H12 CDR Set, comprising;

CDR-L1;

residues 24-34 of SEQ ID NO;

185

CDR-L2;

residues 50-56 of SEQ ID NO;

185 and

CDR-L3;

residues 89-97 of SEQ ID NO;

185

and

VL.2a 38H12 CDR Set, comprising;

CDR-L1;

residues 24-34 of SEQ ID NO;

186

CDR-L2;

residues 50-56 of SEQ ID NO;

186 and

CDR-L3;

residues 89-97 of SEQ ID NO;

186.

3. The binding protein according to claim 2, comprising at least two variable domain CDR sets.

4. The binding protein according to claim 3, wherein the at least two variable domain CDR sets are:

VH 38H12 CDR Set and VL 38H12 CDR Set,

VH 37D10 CDR Set and VL 37D10 CDR Set,

VH 32C7 CDR Set and VL 32C7 CDR Set,

VH 14G1 Set and VL 14G1 CDR Set,

VH 14A11 CDR Set and VL 14A11 CDR Set,

VH 15D6 CDR Set and VL 15D6 CDR Set,

VH.1 38H12 CDR Set and VL.1 38H12 CDR Set,

VH.1 38H12 CDR Set and VL.1a 38H12 CDR Set,

VH.1 38H12 CDR Set and VL.1b 38H12 CDR Set,

VH.1 38H12 CDR Set and VL.2a 38H12 CDR Set,

VH.1a 38H12 CDR Set and VL.1 38H12 CDR Set,

VH.1a 38H12 CDR Set and VL.1a 38H12 CDR Set,

VH.1a 38H12 CDR Set and VL.1b 38H12 CDR Set,

VH.1a 38H12 CDR Set and VL.2a 38H12 CDR Set,

VH.1b 38H12 CDR Set and VL.1 38H12 CDR Set,

VH.1b 38H12 CDR Set and VL.1a 38H12 CDR Set,

VH.1b 38H12 CDR Set and VL.1b 38H12 CDR Set,

VH.1b 38H12 CDR Set and VL.2a 38H12 CDR Set,

VH.2a 38H12 CDR Set and VL.1 38H12 CDR Set,

VH.2a 38H12 CDR Set and VL.1a 38H12 CDR Set,

VH.2a 38H12 CDR Set and VL.1b 38H12 CDR Set, or

VH.2a 38H12 CDR Set and VL.2a 38H12 CDR Set.

5. The binding protein according to claim 1, further comprising a human acceptor framework sequence.

6. The binding protein according to claim 5, wherein the human acceptor framework comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:

6-34 and SEQ ID NOs;

35-98.

7. The binding protein of claim 1, wherein the antigen-binding domain comprises variable domain sequences selected from the group consisting of:

SEQ ID NOs;

157 and 158;

SEQ ID NOs;

161 and 162;

SEQ ID NOs;

163 and 164;

SEQ ID NOs;

165 and 166;

SEQ ID NOs;

167 and 168;

SEQ ID NOs;

169 and 170;

SEQ ID NOs;

179 and 183;

SEQ ID NOs;

179 and 184;

SEQ ID NOs;

179 and 185;

SEQ ID NOs;

179 and 186;

SEQ ID NOs;

180 and 183;

SEQ ID NOs;

180 and 184;

SEQ ID NOs;

180 and 185;

SEQ ID NOs;

180 and 186;

SEQ ID NOs;

181 and 183;

SEQ ID NOs;

181 and 184;

SEQ ID NOs;

181 and 185;

SEQ ID NOs;

181 and 186;

SEQ ID NOs;

182 and 183;

SEQ ID NOs;

182 and 184;

SEQ ID NOs;

182 and 185; and

SEQ ID NOs;

182 and 186.

8. The binding protein according to claim 5, wherein said human acceptor framework comprises at least one framework region amino acid substitution, wherein the amino acid sequence of the framework is at least 65% identical to a sequence of a human germ line acceptor framework and comprises at least 70 amino acid residues identical to the human germline acceptor framework.

9. The binding protein according to claim 1, wherein the binding protein is capable of blocking DLL4 interaction with a Notch protein, modulating a biological function of DLL4, neutralizing a biological function of DLL4, diminishing the ability of DLL4 to bind to its receptor, inhibiting normal angiogenesis, inhibiting VEGFR2 activity, and/or inhibiting VEGFR1 activity.

10. The binding protein according to claim 1, wherein the binding protein has a dissociation constant (K_D) to DLL4 selected from the group consisting of:

at most about 10<sup>−

7</sup> M;

at most about 10<sup>−

8</sup> M;

at most about 10<sup>−

9</sup> M;

at most about 10<sup>−

10</sup> M;

at most about 10<sup>−

11</sup> M;

at most about 10<sup>−

12</sup> M; and

at most about 10<sup>−

13</sup> M.

11. A binding protein construct comprising the binding protein of claim 1, the binding protein construct further comprising a linker polypeptide or an immunoglobulin constant domain.

12. The binding protein construct according to claim 11, wherein the binding protein is selected from the group consisting of:

an immunoglobulin molecule,

a monoclonal antibody,

a chimeric antibody,

a CDR-grafted antibody,

a Fab,

a Fab′

,

a F(ab′

)₂,

an Fv,

a disulfide-linked Fv,

an scFv,

a single domain antibody,

a diabody,

a multispecific antibody,

a dual specific antibody,

a dual variable domain immunoglobulin (DVD-Ig) binding protein, and

a bispecific antibody.

13. The binding protein construct according to claim 11, wherein the binding protein construct comprises a heavy chain immunoglobulin constant domain selected from the group consisting of:

a human IgM constant domain,

a human IgG1 constant domain,

a human IgG2 constant domain,

a human IgG3 constant domain,

a human IgG4 constant domain,

a human IgE constant domain, and

a human IgA constant domain.

14. The binding protein construct according to claim 11, comprising an immunoglobulin constant domain having an amino acid sequence selected from the group consisting of:

SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, and combinations thereof.

15. A binding protein conjugate comprising the binding protein of claim 1, the binding protein conjugate further comprising an agent selected from the group consisting of:

an imaging agent, a therapeutic agent, a cytotoxic agent, and an immunoadhesion molecule.

16. The binding protein conjugate according to claim 15, wherein the agent is an imaging agent selected from the group consisting of a radiolabel, an enzyme, a fluorescent label, a luminescent label, a bioluminescent label, a magnetic label, and biotin.

17. The binding protein conjugate according to claim 16, wherein the imaging agent is a radiolabel selected from the group consisting of:

³H, ¹⁴C, ³⁵S, ⁹⁰Y, ⁹⁹Tc, ¹¹¹In, ¹²⁵I, ¹³¹I, ¹⁷⁷Lu, ¹⁶⁶Ho, and ¹⁵³Sm.

18. The binding protein conjugate according to claim 15, wherein the agent is a therapeutic or cytotoxic agent selected from the group consisting of:

an anti-metabolite, an alkylating agent, an antibiotic, a growth factor, a cytokine, an anti-angiogenic agent, an anti-mitotic agent, an anthracycline, a toxin, and an apoptotic agent.

19. The binding protein according to claim 1, wherein the binding protein possesses a human glycosylation pattern.

20. The binding protein according to claim 1, wherein the binding protein exists as a crystal.

21. An isolated nucleic acid encoding the binding protein amino acid sequence of claim 1.

22. A vector comprising the isolated nucleic acid according to claim 21.

23. The vector according to claim 22, wherein the vector is selected from the group consisting of:

pcDNA, pTT, pTT3, pEFBOS, pBV, pJV, pcDNA3.1 TOPO, pEF6, pHybE, TOPO and pBJ.

24. A host cell comprising the vector of claim 22.

25. The host cell according to claim 24, wherein the host cell is an Escherichia coli cell, a CHO cell, a COS cell, or a yeast cell.

26. A method of producing a binding protein that binds DLL4, comprising culturing the host cell of claim 24 in a culture medium under conditions sufficient to produce a binding protein that binds DLL4.

27. A binding protein produced according to the method of claim 26.

28. A pharmaceutical composition comprising the binding protein of claim 1, and a pharmaceutically acceptable carrier.

29. The pharmaceutical composition of claim 28, further comprising at least one additional agent for treating a disorder in which DLL4 activity is detrimental.

30. The pharmaceutical composition of claim 29, wherein said additional agent is selected from the group consisting of:

a therapeutic agent;

an imaging agent;

an antineoplastic agent;

a chemotherapeutic agent;

an angiogenesis inhibitor;

an anti-VEGF antibody;

an anti-EGFR antibody;

an anti-cMet antibody;

an anti-ErbB3 antibody;

an anti-HER2 antibody;

an anti-CD20 antibody;

aflibercept;

a kinase inhibitor;

a co-stimulation molecule blocker;

an anti-B7.2 antibody;

a CTLA4-Ig;

an adhesion molecule blocker;

an anti-E selectin antibody;

an anti-L selectin antibody;

an anti-cytokine antibody or functional fragment thereof;

an anti-IL-18 antibody;

an anti-TNF antibody;

anti-IL-6 antibody;

methotrexate;

a corticosteroid;

a cyclosporin;

a rapamycin;

FK506;

a DNA alkylating agent;

cisplatin;

carboplatin;

an anti-tubulin agent;

paclitaxel;

docetaxel;

doxorubicin;

gemcitabine;

gemzar;

an anthracycline;

adriamycin;

a topoisiomersase I inhibitor;

a topoisomerase II inhibitor;

5-fluorouracil (5-FU);

leucovorin;

irinotecan;

a receptor tyrosine kinase inhibitor, an apoptosis inhibitor;

a Bcl2/Bclx inhibitor;

erlotinib, gefitinib, a COX-2 inhibitor, celecoxib, cyclosporin;

rapamycin;

a detectable label or reporter molecule;

a TNF antagonist;

an antirheumatic;

a muscle relaxant;

a narcotic;

an analgesic;

an anesthetic;

a sedative;

a local anesthetic;

a neuromuscular blocker;

an antimicrobial agent;

an antipsoriatic agent;

a corticosteroid;

an anabolic steroid;

an erythropoietin;

an immunoglobulin;

an immunosuppressive agent;

a growth hormone;

a hormone replacement drug;

a radiopharmaceutical drug;

an antidepressant;

an antipsychotic drug;

a stimulant;

an asthma medication;

a beta agonist;

an inhaled steroid;

an epinephrine;

an epinephrine analog thereof;

a cytokine; and

a cytokine antagonist.

31. A method for reducing DLL4 activity in a subject, comprising contacting the subject with the binding protein of claim 1 such that DLL4 activity is reduced in the subject.

32. A method of treating a subject for a disease or a disorder in which DLL4 activity is detrimental, comprising administering to the subject the binding protein of claim 1.

33. The method of claim 32, wherein said disorder is selected from the group consisting of:

breast cancer, colon cancer, rectal cancer, lung cancer, oropharynx cancer, hypopharynx cancer, esophageal cancer, stomach cancer, pancreas cancer, liver cancer, gallbladder cancer, bile duct cancer, small intestine cancer, urinary tract cancer, female genital tract cancer, male genital tract cancer, endocrine gland cancer, skin cancer, hemangioma, melanoma, sarcoma, brain tumor, nerve cancer, eye tumor, meninges cancer, solid tumors from hematopoietic malignancy, tumor metastases, ocular neovascularization, edema, rheumatoid arthritis, atherosclerotic plaques, Crohn'"'"'s disease, inflammatory bowel disease, refractory ascites, psoriasis, sarcoidosis, arterial arteriosclerosis, sepsis, peptic ulcers, burns, pancreatitis, polycystic ovarian disease (POD), endometriosis, uterine fibroid, benign prostate hypertrophy, T-cell acute lymphoblastic leukemia (T-ALL), cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), multiple sclerosis (MS), tetralogy of Fallot (TOF), Alagille syndrome (AS), macular degeneration and age-related macular degeneration diseases, and other angiogenesis independent and dependent diseases characterized by aberrant DLL4 activity.

34. The method according to claim 32, wherein the binding protein is administered to the subject by at least one mode selected from the group consisting of:

parenteral, subcutaneous, intramuscular, intravenous, intraarterial, intraarticular, intrabronchial, intraabdominal, intracapsular, intracartilaginous, intracavitary, intracelial, intracerebellar, intracerebroventricular, intracolic, intracervical, intragastric, intrahepatic, intramyocardial, intraosteal, intrapelvic, intrapericardiac, intraperitoneal, intrapleural, intraprostatic, intrapulmonary, intrarectal, intrarenal, intraretinal, intraspinal, intrasynovial, intrathoracic, intrauterine, intravesical, bolus, vaginal, rectal, buccal, sublingual, intranasal, and transdermal.

35. The method according to claim 32, further comprising the step of administering a second agent before, concurrently with, or after administering the binding protein, wherein the second agent is selected from the group consisting of an antibody or fragment thereof capable of binding human VEGFR2;

methotrexate;

an antibody or fragment thereof capable of binding human TNF;

a corticosteroid;

a cyclosporine;

a rapamycin;

FK506;

a non-steroidal anti-inflammatory agent (NSAID);

a radiotherapeutic agent;

an antineoplastic agent;

a chemotherapeutic agent;

a DNA alkylating agent;

cisplatin;

carboplatin;

an anti-tubulin agent;

paclitaxel;

docetaxel;

taxol;

doxorubicin;

gemcitabine;

gemzar;

an anthracycline;

adriamycin;

a topoisomerase I inhibitor;

a topoisomerase II inhibitor;

5-fluorouracil (5-FU);

leucovorin;

irinotecan;

a receptor tyrosine kinase inhibitor;

erlotinib;

gefitinib;

a COX-2 inhibitor;

celecoxib;

a kinase inhibitor;

an angiogenesis inhibitor;

an anti-VEGF antibody;

aflibercept;

a co-stimulation molecule blocker;

an anti-B7.1 antibody;

an anti-B7.2 antibody;

a CTLA4-Ig;

an anti-CD20 antibody;

an adhesion molecule blocker;

an anti-LFA-1 antibody;

an anti-E selectin antibody; and

anti-L selectin antibody;

a small molecule inhibitor;

an anti-cytokine antibody or functional fragment thereof;

an anti-IL-18 antibody;

anti-TNF antibody;

an anti-IL-6 antibody;

an anti-cytokine receptorantibody;

a detectable label or reporter;

a TNF antagonist;

an antirheumatic;

a muscle relaxant;

a narcotic;

an analgesic;

an anesthetic;

a sedative;

a local anesthetic;

a neuromuscular blocker;

an antimicrobial agent;

an antipsoriatic drug;

a corticosteroid;

an anabolic steroid;

an erythropoietin;

an immunization;

an immunoglobulin;

an immunosuppressive agent;

a growth hormone;

a hormone replacement drug;

a radiopharmaceutical drug;

an antidepressant;

an antipsychotic drug;

a stimulant;

an asthma medication;

a beta agonist;

an inhaled steroid;

an epinephrine;

an epinephrine analog;

a cytokine; and

a cytokine antagonist.

36. The binding protein according to claim 1, wherein the binding protein is an antibody.

37. The binding protein according to claim 36, wherein the antibody is selected from the group consisting of a monoclonal antibody, a full-length tetrameric immunoglobulin, an IgG molecule, an IgG1 molecule, a chimeric antibody, a CDR-grafted antibody, a humanized antibody, and an affinity matured antibody.

38. The binding protein according to claim 36, wherein the antibody is a monoclonal antibody.

39. The binding protein according to claim 1, wherein the binding protein is a dual variable domain immunoglobulin (DVD-Ig) binding protein.