Coumarin‐30 Enables Site‐Resolved Detection of Tubulin Ligands by Microscale Thermophoresis

Abstract Tubulins are among the most successful targets for cancer chemotherapy. However, the emergence of drug resistance stimulates the continuous search for novel chemotherapeutics. We discovered that coumarin‐30, a widely available laser dye, binds to the colchicine site of tubulin, inhibiting microtubule dynamics and cancer cell division at submicromolar concentrations. Exploiting the excellent fluorescent properties of coumarin‐30, we developed a fast, accurate, and cost‐effective coumarin‐30‐based microscale thermophoresis (C‐MST) assay as an express method for detecting tubulin–ligand interactions and discriminating colchicine site binders from ligands targeting other protein pockets. Using this assay, we identified several potent tubulin polymerization inhibitors associating with the colchicine site and validated them through in vitro microtubule dynamics and cell cycle assays in cancer cells. Furthermore, the C‐MST assay was demonstrated to detect ligands targeting a novel binding site on tubulin, recently established through crystallographic fragment screening. We confirmed detection of a small‐molecule ligand targeting that site and further designed and characterized a series of its analogs. The ability of the C‐MST assay to detect tubulin binders regardless of their binding site or their effect on microtubule dynamics opens new avenues for developing unconventional modulators of tubulin–tubulin and tubulin–effector interactions thereby facilitating anticancer drug discovery.

Authors

Institutions

Publication Details

Journal
Angewandte Chemie
Published
2025-12-01
DOI
https://doi.org/10.1002/ange.202517086
Primary Topic
Field-Flow Fractionation Techniques
Type
article
Field-Weighted Citation Impact
0.00
Controls
|||
ALL TIME
JAN
FEB
MAR
APR
MAY
JUN
JUL
AUG
SEP
OCT
article

Coumarin‐30 Enables Site‐Resolved Detection of Tubulin Ligands by Microscale Thermophoresis

A. N. Romanov, Vladimir A. Mitkevich, Alexander Makarov, Maksim А. Boichenko et al.
Angewandte Chemie
Field-Flow Fractionation Techniques
article

Coumarin‐30 Enables Site‐Resolved Detection of Tubulin Ligands by Microscale Thermophoresis

A. N. Romanov, Vladimir A. Mitkevich, Alexander Makarov, Maksim А. Boichenko, Vadim V. Mustyatsa, Роман А. Новиков, Mikhail N. Anisimov, Оlga А. Ivanova, Igor V. Trushkov, Andrey Yu. Plodukhin, Ivan A. Vorobjev, Vitaly V. Shorokhov, Иван А. Андреев, Нина К. Ратманова, Nikita B. Gudimchuk, В. А. Тафеенко, Sergey S. Zhokhov, Julia N. Borzunova, Marina Janibekova, Timur P. Tikhonov, Ilya A. Lifshits, Olga I. Kechko, Anton A. Sivachev
article en

Abstract

Abstract Tubulins are among the most successful targets for cancer chemotherapy. However, the emergence of drug resistance stimulates the continuous search for novel chemotherapeutics. We discovered that coumarin‐30, a widely available laser dye, binds to the colchicine site of tubulin, inhibiting microtubule dynamics and cancer cell division at submicromolar concentrations. Exploiting the excellent fluorescent properties of coumarin‐30, we developed a fast, accurate, and cost‐effective coumarin‐30‐based microscale thermophoresis (C‐MST) assay as an express method for detecting tubulin–ligand interactions and discriminating colchicine site binders from ligands targeting other protein pockets. Using this assay, we identified several potent tubulin polymerization inhibitors associating with the colchicine site and validated them through in vitro microtubule dynamics and cell cycle assays in cancer cells. Furthermore, the C‐MST assay was demonstrated to detect ligands targeting a novel binding site on tubulin, recently established through crystallographic fragment screening. We confirmed detection of a small‐molecule ligand targeting that site and further designed and characterized a series of its analogs. The ability of the C‐MST assay to detect tubulin binders regardless of their binding site or their effect on microtubule dynamics opens new avenues for developing unconventional modulators of tubulin–tubulin and tubulin–effector interactions thereby facilitating anticancer drug discovery.

Angewandte ChemieVol. 138(12)
Engelhardt Institute of Molecular Biology (RU), Lomonosov Moscow State University (RU), Semenov Institute of Chemical Physics (RU), N.D. Zelinsky Institute of Organic Chemistry (RU), Center for Theoretical Problems of Physicochemical Pharmacology (RU), Nazarbayev University (KZ)
Openalex Percentile: Top 61%
Field-Flow Fractionation Techniques
DOISourceOpenAlex
AI Navigator

Ask Laika to Summarize, Analyze, and Connect papers live on the map.

Summarize Papers & Methodologies

Extract key findings, datasets, and comparative methods across publications.

Benchmark Rankings & Visual Analytics

Rank top research institutions, authors, funders, topics, and journals by Field-Weighted Citation Impact (FWCI) and paper volume with instant charts.

Connect Distant Disciplines

Bridge topological clusters on the map to find hidden collaborative intersections.