Screening of potential new ROCK inhibitors for future preclinical development

Abstract

Rho kinases (ROCKs) regulate a broad range of cellular processes that involve actin-filament assembly and cell contractility. Therefore, ROCKs are involved in inflammation, angiogenesis and fibrosis. Consequently, ROCK inhibitors could have potential therapeutic effects in several pathologies. Amakem Therapeutics develops selective ROCK inhibitors based on the 'localized drug action' platform with the aim to reduce systemic side effects. The aim of this study was to screen 10 Amakem compounds to select potential new ROCK inhibitors as therapeutic candidates for future development. The stability and cytotoxicity profile of the 10 compounds was first established. Based on these results, 6 compounds were selected for further screening in functional cell based assays. All compounds showed a significant effect on HUVEC and HDF-1 migration. However, only one compound showed an effect on transmigration of HL-60. In the HUVEC proliferation assay a stimulatory effect of bFGF and VEGF was observed. However, there was no effect of the compounds on HUVEC proliferation. In addition, the ROCK inhibitors did not affect angiogenesis of the CAM in vivo. Further experiments are needed to select compounds for future preclinical development.