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

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

Journal

Angewandte Chemie International Edition
ISSN: 1433-7851 , E-ISSN: 1521-3773

Output data

Year: 2025, Article number : e17086, Pages count : DOI: 10.1002/anie.202517086

Authors

Anisimov Mikhail N. ^1,2 , Boichenko Maksim A. ³ , Sivachev Anton A. ³ , Romanov Alexey N. ⁴ , Lifshits Ilya A. ¹ , Borzunova Julia N. ¹ , Janibekova Marina ⁵ , Mustyatsa Vadim V. ⁵ , Tikhonov Timur P. ³ , Plodukhin Andrey Yu. ³ , Shorokhov Vitaly V. ³ , Tafeenko Victor A. ³ , Ratmanova Nina K. ⁶ , Zhokhov Sergey S. ³ , Novikov Roman A. ^6,7 , Kechko Olga I. ⁷ , Makarov Alexander A. ⁷ , Mitkevich Vladimir A. ⁷ , Andreev Ivan A. ⁸ , Vorobjev Ivan A. ^5,8 , Trushkov Igor V. ⁶ , Ivanova Olga A. ³ , Gudimchuk Nikita B. ^1,2

Affiliations

1	Department of Physics M. V. Lomonosov Moscow State University Leninskie Gory 1–2 Moscow 119991 Russia
2	Center for Theoretical Problems of Physicochemical Pharmacology Russian Academy of Sciences Srednyaa Kalitnikovskaya 30 Moscow 109029 Russia
3	Department of Chemistry M. V. Lomonosov Moscow State University Leninskie Gory 1–3 Moscow 119991 Russia
4	N. N. Semenov Institute of Chemical Physics Russian Academy of Sciences Kosygina 4 Moscow 119991 Russia
5	National Laboratory Astana, Kabanbai batyr 53, Astana, 010000 Kazakhstan
6	N. D. Zelinsky Institute of Organic Chemistry Russian Academy of Sciences Moscow 119991 Russia
7	Engelhardt Institute of Molecular Biology Russian Academy of Sciences Vavilova 32 Moscow 119991 Russia
8	Department of Biology School of Sciences and Humanities Nazarbayev University Kabanbai batyr 53 Astana 010000 Kazakhstan

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.

Anisimov M.N. , Boichenko M.A. , Sivachev A.A. , Romanov A.N. , Lifshits I.A. , Borzunova J.N. , Janibekova M. , Mustyatsa V.V. , Tikhonov T.P. , Plodukhin A.Y. , Shorokhov V.V. , Tafeenko V.A. , Ratmanova N.K. , Zhokhov S.S. , Novikov R.A. , Kechko O.I. , Makarov A.A. , Mitkevich V.A. , Andreev I.A. , Vorobjev I.A. , Trushkov I.V. , Ivanova O.A. , Gudimchuk N.B.
Coumarin‐30 Enables Site‐Resolved Detection of Tubulin Ligands by Microscale Thermophoresis
Angewandte Chemie International Edition. 2025. e17086 . DOI: 10.1002/anie.202517086 WOS Scopus OpenAlex

Web of science:	WOS:001628084500001
Scopus:	2-s2.0-105023426015
OpenAlex:	W4416848004

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