Abstract
Using high concentration biochemical assays and fragment-based screening assisted by structure-guided design, we discovered a novel class of Rho-kinase inhibitors. Compound 18 was equipotent for ROCK1 (IC(50) = 650 nM) and ROCK2 (IC(50) = 670 nM), whereas compound 24 was more selective for ROCK2 (IC(50) = 100 nM) over ROCK1 (IC(50) = 1690 nM). The crystal structure of the compound 18-ROCK1 complex revealed that 18 is a type 1 inhibitor that binds the hinge region in the ATP binding site. Compounds 18 and 24 inhibited potently the phosphorylation of the ROCK substrate MLC2 in intact human breast cancer cells.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Adenosine Triphosphate / chemistry
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Binding Sites
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Cardiac Myosins / metabolism
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Cell Line, Tumor
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Crystallography, X-Ray
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Humans
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Mitogen-Activated Protein Kinase 1 / metabolism
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Mitogen-Activated Protein Kinase 3 / metabolism
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Models, Molecular
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Molecular Structure
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Myosin Light Chains / metabolism
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Phosphorylation
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Structure-Activity Relationship
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rho-Associated Kinases / antagonists & inhibitors*
Substances
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Myosin Light Chains
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myosin light chain 2
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Adenosine Triphosphate
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ROCK1 protein, human
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ROCK2 protein, human
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rho-Associated Kinases
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Mitogen-Activated Protein Kinase 1
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Mitogen-Activated Protein Kinase 3
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Cardiac Myosins