A novel small-molecule MRCK inhibitor blocks cancer cell invasion

Unbekandt, M. et al. (2014) A novel small-molecule MRCK inhibitor blocks cancer cell invasion. Cell Communication and Signaling, 12, 54. (doi: 10.1186/s12964-014-0054-x) (PMID:25288205) (PMCID:PMC4195943)

[img] Text
101436.pdf - Published Version
Available under License Creative Commons Attribution.

3MB

Abstract

Background: The myotonic dystrophy kinase-related CDC42-binding kinases MRCKα and MRCKβ regulate actin-myosin contractility and have been implicated in cancer metastasis. Along with the related ROCK1 and ROCK2 kinases, the MRCK proteins initiate signalling events that lead to contractile force generation which powers cancer cell motility and invasion. A potential strategy for cancer therapy is to reduce metastasis by blocking MRCK activity, either alone or in combination with ROCK inhibition. However, to date no potent small molecule inhibitors have been developed with selectivity towards MRCK.<p></p> Results: Screening a kinase-focused small molecule chemical library resulted in the identification of compounds with inhibitory activity towards MRCK. Medicinal chemistry combined with in vitro enzyme profiling led to the discovery of 4-chloro-1-(4-piperidyl)-N-[5-(2-pyridyl)-1H-pyrazol-4-yl]pyrazole-3-carboxamide (BDP00005290; abbreviated as BDP5290) as a potent MRCK inhibitor. X-ray crystallography of the MRCKβ kinase domain in complex with BDP5290 revealed how this ligand interacts with the nucleotide binding pocket. BDP5290 demonstrated marked selectivity for MRCKβ over ROCK1 or ROCK2 for inhibition of myosin II light chain (MLC) phosphorylation in cells. While BDP5290 was able to block MLC phosphorylation at both cytoplasmic actin stress fibres and peripheral cortical actin bundles, the ROCK selective inhibitor Y27632 primarily reduced MLC phosphorylation on stress fibres. BDP5290 was also more effective at reducing MDA-MB-231 breast cancer cell invasion through Matrigel than Y27632. Finally, the ability of human SCC12 squamous cell carcinoma cells to invade a three-dimensional collagen matrix was strongly inhibited by 2 μM BDP5290 but not the identical concentration of Y27632, despite equivalent inhibition of MLC phosphorylation.<p></p> Conclusions: BDP5290 is a potent MRCK inhibitor with activity in cells, resulting in reduced MLC phosphorylation, cell motility and tumour cell invasion. The discovery of this compound will enable further investigations into the biological activities of MRCK proteins and their contributions to cancer progression.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Croft, Dr Daniel and Belshaw, Miss Simone and Crighton, Dr Diane and McConnell, Mrs Patricia and Drysdale, Dr Martin and Schuettelkopf, Dr Alexander and Bower, Dr Justin and Sime, Dr Mairi and Pannifer, Dr Andrew and McArthur, Dr Duncan and Mezna, Dr Mokdad and Unbekandt, Dr Mathieu and Olson, Professor Michael
Authors: Unbekandt, M., Croft, D. R., Crighton, D., Mezna, M., McArthur, D., McConnell, P., Schüttelkopf, A. W., Belshaw, S., Pannifer, A., Sime, M., Bower, J., Drysdale, M., and Olson, M. F.
College/School:College of Medical Veterinary and Life Sciences > Institute of Cancer Sciences
Journal Name:Cell Communication and Signaling
Publisher:BioMed Central
ISSN:1478-811X
ISSN (Online):1478-811X
Published Online:05 October 2014
Copyright Holders:Copyright © 2014 The Authors
First Published:First published in Cell Communication and Signaling 12(54)
Publisher Policy:Reproduced under a Creative Commons License

University Staff: Request a correction | Enlighten Editors: Update this record

Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
576111Development of a novel drug targeting MRCK and ROCKII for the treatment of invasive and/or metastatic cancerMichael OlsonMedical Research Council (MRC)MR/J005126/1ICS - BEATSON INSTITUTE FOR CANCER RES.