Abstract: Microtubules are cytoskeletal components which play important roles in a number of cellular processes. Colchicine binding site inhibitors (CSIs) is one of the major classes of tubulin polymerization inhibitors. Many CSIs have been discovered, designed or synthesized as anticancer agents in the past several years and great progress has been made. We discuss the insights gained so far relevant to the mechanism of CSIs and their common structures. The recent development of CSIs with their biological activity and structure-activity relationship is also reviewed.