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[Cancer Research 57, 4593-4599, October 15, 1997]
© 1997 American Association for Cancer Research
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Humanization of an Anti-Vascular Endothelial Growth Factor Monoclonal Antibody for the Therapy of Solid Tumors and Other Disorders

Leonard G. Presta, Helen Chen, Shane J. O'Connor, Vanessa Chisholm, Y. Gloria Meng, Lynne Krummen, Marjorie Winkler and Napoleone Ferrara1

Departments of Immunology, Process Sciences, Molecular Biology, Bioanalytical Technology and Cardiovascular Research, Genentech, Inc., South San Francisco, California 94080

Vascular endothelial growth factor (VEGF) is a major mediator of angiogenesis associated with tumors and other pathological conditions, including proliferative diabetic retinopathy and age-related macular degeneration. The murine anti-human VEGF monoclonal antibody (muMAb VEGF) A.4.6.1 has been shown to potently suppress angiogenesis and growth in a variety of human tumor cells lines transplanted in nude mice and also to inhibit neovascularization in a primate model of ischemic retinal disease. In this report, we describe the humanization of muMAb VEGF A.4.6.1. by site-directed mutagenesis of a human framework. Not only the residues involved in the six complementarity-determining regions but also several framework residues were changed from human to murine. Humanized anti-VEGF F(ab) and IgG1 variants bind VEGF with affinity very similar to that of the original murine antibody. Furthermore, recombinant humanized MAb VEGF inhibits VEGF-induced proliferation of endothelial cells in vitro and tumor growth in vivo with potency and efficacy very similar to those of muMAb VEGF A.4.6.1. Therefore, recombinant humanized MAb VEGF is suitable to test the hypothesis that inhibition of VEGF-induced angiogenesis is a valid strategy for the treatment of solid tumors and other disorders in humans.

1 To whom requests for reprints should be addressed, at Department of Cardiovascular Research, Genentech, Inc., 460 Point San Bruno Boulevard, South San Francisco, CA 94080. Phone: (415) 225-2968; Fax: (415) 225-6327; E-mail: Ferrara.Napoleone@gene.com.

Received 5/27/97. Accepted 8/16/97.




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 Endocr. Rev.Home page
N. Ferrara
Vascular Endothelial Growth Factor: Basic Science and Clinical Progress
Endocr. Rev., August 1, 2004; 25(4): 581 - 611.
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 The OncologistHome page
M. Cristofanilli and G. N. Hortobagyi
Breast Cancer Highlights: Key Findings from the San Antonio Breast Cancer Symposium: A U.S. Perspective
Oncologist, July 1, 2004; 9(4): 471 - 478.
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 Clin. Cancer Res.Home page
A. B. Sandler, D. H. Johnson, and R. S. Herbst
Anti-Vascular Endothelial Growth Factor Monoclonals in Non-Small Cell Lung Cancer
Clin. Cancer Res., June 15, 2004; 10(12): 4258S - 4262S.
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 NEJMHome page
H. Hurwitz, L. Fehrenbacher, W. Novotny, T. Cartwright, J. Hainsworth, W. Heim, J. Berlin, A. Baron, S. Griffing, E. Holmgren, et al.
Bevacizumab plus Irinotecan, Fluorouracil, and Leucovorin for Metastatic Colorectal Cancer
N. Engl. J. Med., June 3, 2004; 350(23): 2335 - 2342.
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 JCOHome page
D. H. Johnson, L. Fehrenbacher, W. F. Novotny, R. S. Herbst, J. J. Nemunaitis, D. M. Jablons, C. J. Langer, R. F. DeVore III, J. Gaudreault, L. A. Damico, et al.
Randomized Phase II Trial Comparing Bevacizumab Plus Carboplatin and Paclitaxel With Carboplatin and Paclitaxel Alone in Previously Untreated Locally Advanced or Metastatic Non-Small-Cell Lung Cancer
J. Clin. Oncol., June 1, 2004; 22(11): 2184 - 2191.
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 The OncologistHome page
R. S. Herbst and A. B. Sandler
Non-Small Cell Lung Cancer and Antiangiogenic Therapy: What Can Be Expected of Bevacizumab?
Oncologist, June 1, 2004; 9(suppl_1): 19 - 26.
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 The OncologistHome page
H. X. Chen
Expanding the Clinical Development of Bevacizumab
Oncologist, June 1, 2004; 9(suppl_1): 27 - 35.
[Abstract] [Full Text] [PDF]

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 The OncologistHome page
H. S. Rugo
Bevacizumab in the Treatment of Breast Cancer: Rationale and Current Data
Oncologist, June 1, 2004; 9(suppl_1): 43 - 49.
[Abstract] [Full Text] [PDF]

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 Cancer Res.Home page
J. Sun, M. A. Blaskovich, R. K. Jain, F. Delarue, D. Paris, S. Brem, M. Wotoczek-Obadia, Q. Lin, D. Coppola, K. Choi, et al.
Blocking Angiogenesis and Tumorigenesis with GFA-116, a Synthetic Molecule that Inhibits Binding of Vascular Endothelial Growth Factor to its Receptor
Cancer Res., May 15, 2004; 64(10): 3586 - 3592.
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 Clin. Cancer Res.Home page
B. I. Rini, S. Halabi, J. Taylor, E. J. Small, and R. L. Schilsky
Cancer and Leukemia Group B 90206: A Randomized Phase III Trial of Interferon-{alpha} or Interferon-{alpha} Plus Anti-Vascular Endothelial Growth Factor Antibody (Bevacizumab) in Metastatic Renal Cell Carcinoma
Clin. Cancer Res., April 15, 2004; 10(8): 2584 - 2586.
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 Cancer Res.Home page
J. S. Beebe, J. P. Jani, E. Knauth, P. Goodwin, C. Higdon, A. M. Rossi, E. Emerson, M. Finkelstein, E. Floyd, S. Harriman, et al.
Pharmacological Characterization of CP-547,632, a Novel Vascular Endothelial Growth Factor Receptor-2 Tyrosine Kinase Inhibitor for Cancer Therapy
Cancer Res., November 1, 2003; 63(21): 7301 - 7309.
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 Endocr. Rev.Home page
H. E. Turner, A. L. Harris, S. Melmed, and J. A. H. Wass
Angiogenesis in Endocrine Tumors
Endocr. Rev., October 1, 2003; 24(5): 600 - 632.
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 Cancer Res.Home page
B. Ruggeri, J. Singh, D. Gingrich, T. Angeles, M. Albom, H. Chang, C. Robinson, K. Hunter, P. Dobrzanski, S. Jones-Bolin, et al.
CEP-7055: A Novel, Orally Active Pan Inhibitor of Vascular Endothelial Growth Factor Receptor Tyrosine Kinases with Potent Antiangiogenic Activity and Antitumor Efficacy in Preclinical Models
Cancer Res., September 15, 2003; 63(18): 5978 - 5991.
[Abstract] [Full Text] [PDF]

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 Diabetes CareHome page
T. A. Ciulla, A. G. Amador, and B. Zinman
Diabetic Retinopathy and Diabetic Macular Edema: Pathophysiology, screening, and novel therapies
Diabetes Care, September 1, 2003; 26(9): 2653 - 2664.
[Abstract] [Full Text] [PDF]

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 NEJMHome page
J. C. Yang, L. Haworth, R. M. Sherry, P. Hwu, D. J. Schwartzentruber, S. L. Topalian, S. M. Steinberg, H. X. Chen, and S. A. Rosenberg
A Randomized Trial of Bevacizumab, an Anti-Vascular Endothelial Growth Factor Antibody, for Metastatic Renal Cancer
N. Engl. J. Med., July 31, 2003; 349(5): 427 - 434.
[Abstract] [Full Text] [PDF]

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 Clin. Cancer Res.Home page
F. Ciardiello, R. Caputo, V. Damiano, R. Caputo, T. Troiani, D. Vitagliano, F. Carlomagno, B. M. Veneziani, G. Fontanini, A. R. Bianco, et al.
Antitumor Effects of ZD6474, a Small Molecule Vascular Endothelial Growth Factor Receptor Tyrosine Kinase Inhibitor, with Additional Activity against Epidermal Growth Factor Receptor Tyrosine Kinase
Clin. Cancer Res., April 1, 2003; 9(4): 1546 - 1556.
[Abstract] [Full Text] [PDF]

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 Mol. Pathol.Home page
E S Bamberger and C W Perrett
Angiogenesis in epithelian ovarian cancer
Mol. Pathol., December 1, 2002; 55(6): 348 - 359.
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 Cancer Res.Home page
M. Hagedorn, L. Zilberberg, J. Wilting, X. Canron, G. Carrabba, C. Giussani, M. Pluderi, L. Bello, and A. Bikfalvi
Domain Swapping in a COOH-terminal Fragment of Platelet Factor 4 Generates Potent Angiogenesis Inhibitors
Cancer Res., December 1, 2002; 62(23): 6884 - 6890.
[Abstract] [Full Text] [PDF]

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 Cancer Res.Home page
E. Barbera-Guillem, J. K. Nyhus, C. C. Wolford, C. R. Friece, and J. W. Sampsel
Vascular Endothelial Growth Factor Secretion by Tumor-infiltrating Macrophages Essentially Supports Tumor Angiogenesis, and IgG Immune Complexes Potentiate the Process
Cancer Res., December 1, 2002; 62(23): 7042 - 7049.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
N. Gao, B.-H. Jiang, S. S. Leonard, L. Corum, Z. Zhang, J. R. Roberts, J. Antonini, J. Z. Zheng, D. C. Flynn, V. Castranova, et al.
p38 Signaling-mediated Hypoxia-inducible Factor 1alpha and Vascular Endothelial Growth Factor Induction by Cr(VI) in DU145 Human Prostate Carcinoma Cells
J. Biol. Chem., November 15, 2002; 277(47): 45041 - 45048.
[Abstract] [Full Text] [PDF]

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 Clin. Cancer Res.Home page
W. D. Fox, B. Higgins, K. M. Maiese, M. Drobnjak, C. Cordon-Cardo, H. I. Scher, and D. B. Agus
Antibody to Vascular Endothelial Growth Factor Slows Growth of an Androgen-independent Xenograft Model of Prostate Cancer
Clin. Cancer Res., October 1, 2002; 8(10): 3226 - 3231.
[Abstract] [Full Text] [PDF]

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 Proc. Natl. Acad. Sci. USAHome page
E. S. Kim, A. Serur, J. Huang, C. A. Manley, K. W. McCrudden, J. S. Frischer, S. Z. Soffer, L. Ring, T. New, S. Zabski, et al.
Potent VEGF blockade causes regression of coopted vessels in a model of neuroblastoma
PNAS, August 20, 2002; 99(17): 11399 - 11404.
[Abstract] [Full Text] [PDF]

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 Cancer Res.Home page
K. M. Comerford, T. J. Wallace, J. Karhausen, N. A. Louis, M. C. Montalto, and S. P. Colgan
Hypoxia-inducible Factor-1-dependent Regulation of the Multidrug Resistance (MDR1) Gene
Cancer Res., June 1, 2002; 62(12): 3387 - 3394.
[Abstract] [Full Text] [PDF]

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 Cancer Res.Home page
C. L. Reimer, N. Agata, J. G. Tammam, M. Bamberg, W. M. Dickerson, G. D. Kamphaus, S. L. Rook, M. Milhollen, R. Fram, R. Kalluri, et al.
Antineoplastic Effects of Chemotherapeutic Agents Are Potentiated by NM-3, an Inhibitor of Angiogenesis
Cancer Res., February 1, 2002; 62(3): 789 - 795.
[Abstract] [Full Text] [PDF]

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 Cold Spring Harb Symp Quant BiolHome page
J. LECOUTER, R. LIN, and N. FERRARA
The Role of EG-VEGF in the Regulation of Angiogenesis in Endocrine Glands
Cold Spring Harb Symp Quant Biol, January 1, 2002; 67(0): 217 - 222.
[Abstract] [PDF]

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 The OncologistHome page
F. J. Giles
The Vascular Endothelial Growth Factor (VEGF) Signaling Pathway: A Therapeutic Target in Patients with Hematologic Malignancies
Oncologist, October 1, 2001; 6(2008): 32 - 39.
[Abstract] [Full Text] [PDF]

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C. Wulff, H. Wilson, J. S. Rudge, S. J. Wiegand, S. F. Lunn, and H. M. Fraser
Luteal Angiogenesis: Prevention and Intervention by Treatment with Vascular Endothelial Growth Factor TrapA40
J. Clin. Endocrinol. Metab., July 1, 2001; 86(7): 3377 - 3386.
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 Am. J. Physiol. Cell Physiol.Home page
N. Ferrara
Role of vascular endothelial growth factor in regulation of physiological angiogenesis
Am J Physiol Cell Physiol, June 1, 2001; 280(6): C1358 - C1366.
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 Cancer Res.Home page
Q. Shi, X. Le, J. L. Abbruzzese, Z. Peng, C.-N. Qian, H. Tang, Q. Xiong, B. Wang, X.-C. Li, and K. Xie
Constitutive Sp1 Activity Is Essential for Differential Constitutive Expression of Vascular Endothelial Growth Factor in Human Pancreatic Adenocarcinoma
Cancer Res., May 1, 2001; 61(10): 4143 - 4154.
[Abstract] [Full Text]

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 JEMHome page
G. T. Furuta, J. R. Turner, C. T. Taylor, R. M. Hershberg, K. Comerford, S. Narravula, D. K. Podolsky, and S. P. Colgan
Hypoxia-inducible Factor 1-dependent Induction of Intestinal Trefoil Factor Protects Barrier Function during Hypoxia
J. Exp. Med., April 30, 2001; 193(9): 1027 - 1034.
[Abstract] [Full Text] [PDF]

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 JCBHome page
B. R. Zetter
Hold that Line: Angiomotin Regulates Endothelial Cell Motility
J. Cell Biol., March 19, 2001; 152(6): F35 - F36.
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 FASEB J.Home page
H.-Q. MIAO, P. LEE, H. LIN, S. SOKER, and M. KLAGSBRUN
Neuropilin-1 expression by tumor cells promotes tumor angiogenesis and progression
FASEB J, December 1, 2000; 14(15): 2532 - 2539.
[Abstract] [Full Text]

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 Cancer Res.Home page
H.-P. Gerber, J. Kowalski, D. Sherman, D. A. Eberhard, and N. Ferrara
Complete Inhibition of Rhabdomyosarcoma Xenograft Growth and Neovascularization Requires Blockade of Both Tumor and Host Vascular Endothelial Growth
Cancer Res., November 1, 2000; 60(22): 6253 - 6258.
[Abstract] [Full Text] [PDF]


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