Development and Preclinical Characterization of a Humanized Antibody Targeting CXCL12

• Published in: Clinical cancer research Vol. 19; no. 16; pp. 4433 - 4445
• Main Authors: CUILING ZHONG, JIANYONG WANG, QUINTANA, Leah, GRIBLING, Peter, SUTO, Eric, BARCK, Kai, CORPUZ, Racquel, YAO, Jenny, TAKKAR, Rashi, LEE, Wyne P, DAMICO-BEYER, Lisa A, CARANO, Richard D, BING LI, ADAMS, Camellia, KELLEY, Robert F, WEIRU WANG, FERRARA, Napoleone, HONG XIANG, ULTSCH, Mark, COONS, Mary, WONG, Terence, CHIANG, Nancy Y, CLARK, Suzy, CLARK, Robyn
• Format: Journal Article
• Language: English
• Published: Philadelphia, PA American Association for Cancer Research 15.08.2013
• Subjects: Angiogenesis Inhibitors - administration & dosage; Angiogenesis Inhibitors - chemistry; Angiogenesis Inhibitors - pharmacology; Animals; Anti-Inflammatory Agents - administration & dosage; Anti-Inflammatory Agents - chemistry; Anti-Inflammatory Agents - pharmacology; Antibodies, Monoclonal - pharmacology; Antibodies, Monoclonal, Humanized - administration & dosage; Antibodies, Monoclonal, Humanized - chemistry; Antibodies, Monoclonal, Humanized - pharmacology; Antineoplastic agents; Antineoplastic Agents - administration & dosage; Antineoplastic Agents - chemistry; Antineoplastic Agents - pharmacology; Arthritis, Experimental - drug therapy; Arthritis, Experimental - metabolism; Biological and medical sciences; Cell Line, Tumor; Chemokine CXCL12 - antagonists & inhibitors; Chemokine CXCL12 - chemistry; Chemokine CXCL12 - metabolism; Cricetinae; Disease Models, Animal; Drug Evaluation, Preclinical; Drug Synergism; Epitope Mapping; Female; General aspects; Humans; Medical sciences; Mice; Models, Molecular; Neoplasm Metastasis; Neoplasms - drug therapy; Neoplasms - pathology; Pharmacology. Drug treatments; Protein Conformation; Tumor Burden - drug effects; Vascular Endothelial Growth Factor A - antagonists & inhibitors; Xenograft Model Antitumor Assays; Chemokine CXCL12; Rodentia; Preclinical study; Monoclonal antibody; Malignant tumor; In vitro; Biological activity; Inflammatory disease; In vivo; Vertebrata; Mammalia; Animal; Humanized antibody; Hamster; Cancer
• ISSN: 1078-0432; 1557-3265
• DOI: 10.1158/1078-0432.ccr-13-0943

Our goal was to develop a potent humanized antibody against mouse/human CXCL12. This report summarized its in vitro and in vivo activities. Cell surface binding and cell migration assays were used to select neutralizing hamster antibodies, followed by testing in several animal models. Monoclonal antibody (mAb) 30D8 was selected for humanization based on its in vitro and in vivo activities. 30D8, a hamster antibody against mouse and human CXCL12α, CXCL12β, and CXCL12γ, was shown to dose-dependently block CXCL12α binding to CXCR4 and CXCR7, and CXCL12α-induced Jurkat cell migration in vitro. Inhibition of primary tumor growth and/or metastasis was observed in several models. 30D8 alone significantly ameliorated arthritis in a mouse collagen-induced arthritis model (CIA). Combination with a TNF-α antagonist was additive. In addition, 30D8 inhibited 50% of laser-induced choroidal neovascularization (CNV) in mice. Humanized 30D8 (hu30D8) showed similar in vitro and in vivo activities as the parental hamster antibody. A crystal structure of the hu30D8 Fab/CXCL12α complex in combination with mutational analysis revealed a "hot spot" around residues Asn(44)/Asn(45) of CXCL12α and part of the RFFESH region required for CXCL12α binding to CXCR4 and CXCR7. Finally, hu30D8 exhibited fast clearance in cynomolgus monkeys but not in rats. CXCL12 is an attractive target for treatment of cancer and inflammation-related diseases; hu30D8 is suitable for testing this hypothesis in humans.