title: Biochemical characterization and structure determination of a potent, selective antibody inhibitor of human MMP9.
abstract: Matrix metalloproteinase 9 (MMP9) is a member of a large family of proteases that are secreted as inactive zymogens. It is a key regulator of the extracellular matrix, involved in the degradation of various extracellular matrix proteins. MMP9 plays a pathological role in a variety of inflammatory and oncology disorders and has long been considered an attractive therapeutic target. GS-5745, a potent, highly selective humanized monoclonal antibody inhibitor of MMP9, has shown promise in treating ulcerative colitis and gastric cancer. Here we describe the crystal structure of GS-5745·MMP9 complex and biochemical studies to elucidate the mechanism of inhibition of MMP9 by GS-5745. GS-5745 binds MMP9 distal to the active site, near the junction between the prodomain and catalytic domain, and inhibits MMP9 by two mechanisms. Binding to pro-MMP9 prevents MMP9 activation, whereas binding to active MMP9 allosterically inhibits activity.
authors.affiliation: From Gilead Sciences, Inc., Foster City, California 94404.
authors.forename: Todd C
authors.initials: TC
authors.lastname: Appleby
authors.affiliation: From Gilead Sciences, Inc., Foster City, California 94404.
authors.forename: Andrew E
authors.initials: AE
authors.lastname: Greenstein
authors.affiliation: From Gilead Sciences, Inc., Foster City, California 94404.
authors.forename: Magdeleine
authors.initials: M
authors.lastname: Hung
authors.affiliation: From Gilead Sciences, Inc., Foster City, California 94404.
authors.forename: Albert
authors.initials: A
authors.lastname: Liclican
authors.affiliation: From Gilead Sciences, Inc., Foster City, California 94404.
authors.forename: Maile
authors.initials: M
authors.lastname: Velasquez
authors.affiliation: From Gilead Sciences, Inc., Foster City, California 94404.
authors.forename: Armando G
authors.initials: AG
authors.lastname: Villaseñor
authors.affiliation: From Gilead Sciences, Inc., Foster City, California 94404.
authors.forename: Ruth
authors.initials: R
authors.lastname: Wang
authors.affiliation: From Gilead Sciences, Inc., Foster City, California 94404.
authors.forename: Melanie H
authors.initials: MH
authors.lastname: Wong
authors.affiliation: From Gilead Sciences, Inc., Foster City, California 94404.
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Liu
authors.affiliation: From Gilead Sciences, Inc., Foster City, California 94404.
authors.forename: Giuseppe A
authors.initials: GA
authors.lastname: Papalia
authors.affiliation: From Gilead Sciences, Inc., Foster City, California 94404.
authors.forename: Brian E
authors.initials: BE
authors.lastname: Schultz
authors.affiliation: From Gilead Sciences, Inc., Foster City, California 94404.
authors.forename: Roman
authors.initials: R
authors.lastname: Sakowicz
authors.affiliation: From Gilead Sciences, Inc., Foster City, California 94404.
authors.forename: Victoria
authors.initials: V
authors.lastname: Smith
authors.affiliation: From Gilead Sciences, Inc., Foster City, California 94404 HyockJoo.Kwon@gilead.com.
authors.forename: Hyock Joo
authors.initials: HJ
authors.lastname: Kwon
mesh_terms: D000495:Allosteric Site; D000906:Antibodies / Q000737:chemistry; D061067:Antibodies, Monoclonal, Humanized / Q000737:chemistry*; D020134:Catalytic Domain; D003093:Colitis, Ulcerative / Q000188:drug therapy*; D018360:Crystallography, X-Ray; D015195:Drug Design; D004353:Drug Evaluation, Preclinical; D005780:Gelatin / Q000737:chemistry; D017353:Gene Deletion; D057809:HEK293 Cells; D006801:Humans; D020128:Inhibitory Concentration 50; D020780:Matrix Metalloproteinase 9 / Q000737:chemistry*; D061965:Matrix Metalloproteinase Inhibitors / Q000737:chemistry*; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry; D013274:Stomach Neoplasms / Q000188:drug therapy*; D020349:Surface Plasmon Resonance
chemical_list: D000906:Antibodies; D061067:Antibodies, Monoclonal, Humanized; D061965:Matrix Metalloproteinase Inhibitors; D011994:Recombinant Proteins; D005780:Gelatin; C579270:MMP9 protein, human; D020780:Matrix Metalloproteinase 9
keywords: X-ray crystallography; allosteric regulation; antibody; enzyme inhibitor; matrix metalloproteinase (MMP)