Colchicine is a therapeutic agent currently used in therapies of many diseases. It also shows antimitotic effects, and its high cytotoxic activity against different cancer cell lines has been demonstrated many times. To overcome the limitations of colchicine use in anticancer therapy, we synthesized a series of novel triple-modified 4-chloro-7-carbamatethiocolchicines. All the synthesized compounds have been tested in vitro to evaluate their cytotoxicity toward A549, MCF-7, LoVo, LoVo/DX, and BALB/3T3 cell lines. Additionally, their mode of binding to beta-tubulin was evaluated in silico. The majority of triple-modified colchicine derivatives exhibited significantly higher cytotoxicity than colchicine, doxorubicin, and cisplatin against tested cancerous cell lines with much higher selectivity index values for four of them. CI - (c) 2019 John Wiley & Sons Ltd.