Comparative Analysis of Small-Molecule LIMK1/2 Inhibitors: Chemical Synthesis, Biochemistry, and Cellular Activity

LIM domain kinases 1 and 2 (LIMK1 and LIMK2) are crucial regulators of actin dynamics, affecting key cellular processes such as proliferation and migration. By phosphorylating and inactivating cofilin, LIMK1 and LIMK2 promote actin polymerization. Consequently, LIMK inhibitors are gaining attention as potential treatments for various cancers and neurological disorders. To advance the understanding of LIMK function in health and disease, high-quality chemical probes are essential. In this study, we conducted a comparative analysis of 17 LIMK1/2 inhibitors using a range of in vitro enzymatic and cellular assays. Our evaluation identified three compounds—TH-257, LIJTF500025, and LIMKi3—as highly potent and selective inhibitors. These compounds are now proposed as valuable pharmacological tools for studying LIMK function in both in vitro and in vivo settings.