Slingshot-2 (SSH-2), one of 61 dual specificity phosphatases (DUSPs), has been shown to be associated with the progression of cancer and Alzheimer's disease by other studies. This DUSP activates cofilin through dephosphorylation and serves to regulate several cell functions, including growth and movement. Finding a specific inhibitor for SSH-2 may have profound impact in medicine because of its disease association. This study executed a large-scale virtual screening experiment in an attempt to find specific inhibitors for SSH-2 from a small molecule, chemical database containing over 2 million compounds, called ZINC. Results from 23 DUSP screenings, specifically SSH-2, Cdc14B, Cdc25A, Cdc25b, DUSP18, JSP-1, KAP, MKP-3, MKP-4, MKP-5, MKP-6, MKP-8, MTMR2, PAC-1, PTEN, PRL-1, PRL-3, TMDP, VHR, VHY, VHZ, VH1, and VH3, were used to identify specific inhibitors from the large ZINC database. Eleven compounds were identified as potential specific inhibitors for SSH-2 with a disparity mean of at least 8000 and standard deviation less than 7000.
Preliminary testing on the effectiveness of three compounds, ZINC03377116, ZINC04307500, and ZINC05501214, from the list of 11 identified compounds was investigated in vitro against HeLa cells. Cells treated with ZINC03377116 and ZINC06601214 showed a decrease in actin filaments. This indicates that these compounds did not inhibit the activity of SSH-2. However, cells treated with ZINC05501214 showed an increase in actin stress fibers at low compound concentrations.