The aim of the project is the development of selective myosin inhibitors targeting non-muscle myosin-2 (NM2) isoforms, which play essential roles in cell division, cell movement and cell shape determination, thereby regulating processes such as neuronal plasticity. In our earlier project we identified five NM2 specific tool compounds by high-content screening, which molecules selectively inhibited NM2 isofoms, while skeletal, cardiac and smooth muscle myosin 2 isoforms remained unaffected. In the current project the most efficient tool compound will be further optimized to a lead compound, as the next level of drug development. In the first half year of the project we will synthesize and isolate 130 individual molecules in mg scale. Furthermore, we determine the inhibitory properties (IC50, maximal inhibition) of each molecule on the six human myosin-2 isoforms, which provides the basis for the detailed structure-activity-relationship (SAR) analysis. The best 20 inhibitors will be tested on primary neurons for neurite outgrowth inducing potentials. Moreover, we determine the mutagenicity and cytotoxicity of the most promising inhibitors. Since the binding site of the new compound-family has not been identified on myosins, we will apply dynamic blind docking methods to determine the possible binding sites, which results will significantly contribute to the understanding of their mechanism of inhibition and will also accelerate the development of more efficient inhibitors.

Máté Gyimesi, Shadar Kumar, István Lőrincz

Result_May 2020