title: Identification of novel ROCK inhibitors with anti-migratory and anti-invasive activities.
abstract: ROCK1 and ROCK2 mediate important processes such as cell migration, invasion and metastasis, making them good targets for the development of antitumor agents. Recently, using a fragment-based approach and X-ray crystallography, we reported on the design and synthesis of novel Rho-kinase inhibitors (RKIs). Here, we selected a pair of RKIs, the closely related structural analogs RKI-18 (potent; IC50 values of 397 nM (ROCK1) and 349 nM (ROCK2)) and RKI-11 (weak/inactive; IC50 values of 38 μM (ROCK1) and 45 μM (ROCK2)), as chemical probes and determined their effects on cytoskeleton organization, signaling, apoptosis, anchorage-dependent and independent growth, migration and invasion. RKI-18 but not RKI-11 suppresses potently the phosphorylation of the ROCK substrate myosin light chain 2 (MLC2) in intact human breast, lung, colon and prostate cancer cells. Furthermore, RKI-18 is highly selective at decreasing the levels of P-MLC2 over those of P-Akt, P-S6 and P-Erk ½. RKI-18 suppresses ROCK-mediated actin fiber formation, following stimulation with LPA as well as p21-activated kinase (PAK)-mediated lamellipodia and filopodia formation following bradykinin or PDGF stimulation. Furthermore, RKI-18 but not RKI-11 inhibits migration, invasion and anchorage-independent growth of human breast cancer cells. The fact that the active ROCK inhibitor RKI-18, but not the inactive closely related structural analog RKI-11 is effective at suppressing malignant transformation suggests that inhibition of ROCK with RKI-18 results in preventing migration, invasion and anchorage-independent growth. The potential of this class of RKIs as anti-tumor agents warrants further advanced preclinical studies.
authors: Drug Discovery Department, H Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA.
authors: R A
authors: RA
authors: Patel
authors: Drug Discovery Department, H Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA.
authors: Y
authors: Y
authors: Liu
authors: Drug Discovery Department, H Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA.
authors: B
authors: B
authors: Wang
authors: Drug Discovery Department, H Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA.
authors: R
authors: R
authors: Li
authors: 1] Drug Discovery Department, H Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA [2] Departments of Oncologic Sciences and Molecular Medicine, University of South Florida, Tampa, FL, USA.
authors: S M
authors: SM
authors: Sebti
mesh_terms: D016475:3T3 Cells; D000199:Actins / Q000378:metabolism; D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D024722:Cardiac Myosins / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects*; D003599:Cytoskeleton / Q000187:drug effects; D006801:Humans; D051379:Mice; D018994:Myosin Light Chains / Q000378:metabolism; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D009369:Neoplasms / Q000188:drug therapy / Q000201:enzymology; D010766:Phosphorylation / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects; D054462:p21-Activated Kinases / Q000378:metabolism; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D000199:Actins; D000970:Antineoplastic Agents; D018994:Myosin Light Chains; D047428:Protein Kinase Inhibitors; C107184:myosin light chain 2; C515851:ROCK1 protein, human; C515852:ROCK2 protein, human; D054462:p21-Activated Kinases; D054460:rho-Associated Kinases; D024722:Cardiac Myosins
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