Discovery of a novel microtubule destabilizing agent targeting the colchicine site based on molecular docking.

in Biochemical pharmacology by Jiangying Cai, Miao He, Yingying Wang, Hui Zhang, Yaxin Xu, Yubin Wang, Chongge You, Hongwei Gao

TLDR

  • Scientists discovered a new medicine, C10, that has potential to fight cancer by targeting microtubules in tumor cells.
  • C10 was found to be effective at inhibiting tumor cell growth, reducing migratory and invasive ability, and promoting apoptosis.

Abstract

Although a number of microtubule-targeting agents have been used in tumour therapy, their resistance and drug toxicity pose clinical challenges. Microtubule destabilizing agents (MDAs) targeting the colchicine site were found to have the advantage of being able to overcome drug resistance. In our previous studies, we identified a novel MDA from the compound database based on virtual screening methods. Its chemical formula is CHNOS, abbreviated as C10. In this study, molecular docking methods confirmed that the binding pattern of C10 to tubulin is similar to that of colchicine. Immunofluorescence staining and tubulin polymerization experiments showed that C10 disrupts the microtubule network and reduces the polymerization efficiency of tubulin. Cell proliferation and toxicity assay showed that C10 could effectively inhibit the growth of tumour cells. After 72 h treatment, the semi-inhibitory concentrations of A549, MCF-7 and HepG2 were 18.83 μM, 16.32 μM and 16.92 μM. Colony formation assay and EdU staining also showed that C10 significantly inhibited the proliferative capacity of tumour cells. Meanwhile, it was found by wound healing and transwell assay that the migration and invasive ability of tumour cells were relatively weakened after treatment with C10. Furthermore, flow cytometry analysis and western blot revealed that C10 reduced the expression of the B-cell lymphoma 2 (BCL-2) and upregulated the level of the Bcl-2-associated X protein (BAX), which in turn activated caspase-3 to promote apoptosis. Finally, the vivo studies in animal showed that C10 significantly inhibited the growth of tumour in nude mice without significant drug toxicity. Thus, C10 may be a colchicine binding site inhibitor with anticancer potential. Abbreviations: BAX, bcl-2-associated X protein; BCL-2, the B-cell lymphoma 2; BSA, bovine albumin; CBSIs, colchicine binding site inhibitors; CCK-8, cell counting kits-8; DAPI, 4',6-diamidino-2-phenylindole; DMSO, dimethyl sulfoxide; EdU, 5-ethynyl-2'-deoxyuridine; FITC, fluorescein Isothiocyanate; GAPDH,glyceraldehyde-3-phosphate dehydrogenase; H-E, hematoxylin-eosin; HRP, horseradish Peroxidase; IHC, immunohistochemistry; MDAs, microtubule destabilizing agents; MSAs, microtubule stabilizing agents; MTAs, microtubule-targeting agents; PBS, phosphate buffered saline; PI, propidium Iodide; PVDF, polyvinylidenefluoride.

Overview

  • The study aimed to evaluate the anticancer potential of a novel microtubule-targeting agent, C10, which binds to the colchicine site of tubulin.
  • The study used a combination of in vitro and in vivo assays to evaluate C10's ability to inhibit tumor cell growth, proliferation, migration, and invasion, and its effects on apoptosis and toxicology.
  • The primary objective of the study was to identify C10 as a potential agent for cancer therapy, which may overcome the limitations of existing microtubule-targeting agents.

Comparative Analysis & Findings

  • C10 was found to disrupt the microtubule network and reduce the polymerization efficiency of tubulin, leading to the inhibition of tumor cell growth and proliferation.
  • C10 effectively inhibited the growth of tumor cells, with semi-inhibitory concentrations ranging from 16.32 to 18.83 μM, and significantly reduced the expression of BCL-2 and upregulated the level of BAX.
  • C10 reduced the migratory and invasive ability of tumor cells, and activated caspase-3 to promote apoptosis.

Implications and Future Directions

  • The study suggests that C10 may be a promising agent for cancer therapy, which can overcome the limitations of existing microtubule-targeting agents.
  • Further studies are needed to evaluate C10's efficacy and safety in human clinical trials, as well as to understand its mechanisms of action and potential combination therapies.
  • Elucidating the structure-activity relationship of C10 and its analogues may lead to the development of more potent and selective microtubule-targeting agents.
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