MS-444 suppresses the activity of purified smooth muscle myosin light chain kinase (MLCK) (IC50: 10 μM).

MS-444 is a small molecule RNA-binding protein HuR (ELAVL1) inhibitor. Colorectal cancer (CRC) cells that show HuR overexpression are treated with MS-444 (1-100 μM) for 48 hr (IC50s: 10.98±1.76 μM, 12.84±2.10 μM, 5.60±0.90 μM, 14.21±2.11 μM, and 10.98±1.24 μM for HCT116, HCA-7, RKO, HT-29, and SW480 cells, respectively). Contrasting effects are observed using non-transformed small intestinal (RIE-1 (IC50=40.70±3.53 μM)) and colonic (YAMC (IC50=28.16±3.23 μM)) epithelial cells. Both non-transformed cell lines are ~3- to 4-fold less responsive to MS-444-mediated growth inhibition, with IC50 values of 40.70 μM and 28.16 μM (P<0.05). Growth inhibition is observed in all CRC lines with IC50 values ranging from 5.60 μM to 14.21 μM with observable effects seen at 10 μM MS-444 [2].

Mice bearing HCT116 cell xenografts receive IP injections of MS-444 (25 mg/kg bw) or vehicle every 48 hr to test the effects of MS-444 on CRC cell growth in vivo. Over the experiment course, mice do not show any adverse effects and maintained similar weights. Anti-tumor effects of MS-444 are observed with an approximately 1.7-fold reduction in tumor size. Mice treated with MS-444 shows a marked 2- to 3-fold reduction in microvessel density (MVD), showing the anti-angiogenic potential of MS-444[2].