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Biochemical characterization and structure determination of a potent, selective antibody inhibitor of human MMP9
Todd C Appleby1, Andrew E Greenstein1, Magdeleine Hung1
1From Gilead Sciences, Inc., Foster City, California 94404.
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.
Insights
GS-5745, an antibody targeting matrix metalloproteinase 9 (MMP9), inhibits MMP9 by blocking its activation and activity. This dual mechanism offers a promising therapeutic strategy for MMP9-related diseases.
Area of Science:
- Biochemistry
- Structural Biology
- Immunology
Background:
- Matrix metalloproteinase 9 (MMP9) is a protease regulating the extracellular matrix.
- MMP9 is implicated in inflammatory and oncology disorders, making it a therapeutic target.
- GS-5745 is a selective antibody inhibitor of MMP9 with therapeutic potential.
Purpose of the Study:
- To elucidate the mechanism of MMP9 inhibition by GS-5745.
- To determine the crystal structure of the GS-5745·MMP9 complex.
- To understand how GS-5745 interacts with MMP9 to inhibit its activity.
Main Methods:
- X-ray crystallography to determine the complex structure.
- Biochemical assays to study MMP9 inhibition.
- Analysis of GS-5745 binding site and its effect on MMP9 activity.
Main Results:
- The crystal structure of the GS-5745·MMP9 complex was determined.
- GS-5745 binds to MMP9 near the prodomain-catalytic domain junction, away from the active site.
- GS-5745 inhibits MMP9 through dual mechanisms: preventing activation of pro-MMP9 and allosterically inhibiting active MMP9.
Conclusions:
- GS-5745 employs a unique dual mechanism to inhibit MMP9.
- The binding site and inhibitory mechanisms elucidated provide insights for developing targeted MMP9 therapies.
- GS-5745 represents a promising therapeutic agent for diseases involving MMP9.
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