High-affinity ligands of the colchicine domain in tubulin based on a structure-guided design
| Autor: | Eva-María Priego, Michel O. Steinmetz, María-Jesús Pérez-Pérez, Juan Estévez Gallego, María-José Camarasa, Asier Gómez-SanJuan, Sandra Liekens, Solange Martins, Isabel Barasoain, J. Fernando Díaz, Federico Gago, Andrea E. Prota, Oskía Bueno |
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| Přispěvatelé: | Ministerio de Economía, Industria y Competitividad (España), Swiss National Science Foundation, European Commission |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
lcsh:Medicine Plasma protein binding Ligands 01 natural sciences Article Cell Line 03 medical and health sciences chemistry.chemical_compound Cell Movement Tubulin Cell Line Tumor 0103 physical sciences Humans Colchicine Binding site lcsh:Science Binding Sites Multidisciplinary 010304 chemical physics biology Cyclohexanones Ligand lcsh:R Endothelial Cells Binding constant Tubulin Modulators In vitro Molecular Docking Simulation 030104 developmental biology chemistry Cell culture biology.protein Biophysics lcsh:Q Protein Binding |
| Zdroj: | Digital.CSIC. Repositorio Institucional del CSIC instname Scientific Reports, Vol 8, Iss 1, Pp 1-17 (2018) Repositorio Institucional de la Consejería de Sanidad de la Comunidad de Madrid Consejería de Sanidad de la Comunidad de Madrid Scientific Reports |
| Popis: | Microtubule-targeting agents that bind at the colchicine-site of tubulin are of particular interest in antitumoral therapy due to their dual mechanism of action as antimitotics and vascular disrupting agents. Cyclohexanediones derivatives have been described as a new family of colchicine-domain binders with an association constant to tubulin similar to that of colchicine. Here, the highresolution structures of tubulin in complex with cyclohexanediones TUB015 and TUB075 were solved by X-ray crystallography. A detailed analysis of the tubulin-TUB075 interaction by means of computational affinity maps allowed the identification of two additional regions at the binding site that were addressed with the design and synthesis of a new series of cyclohexanediones with a distal 2-substituted benzofurane. These new compounds showed potent antiproliferative activity with IC50 values in the nM range, arrested cell cycle progression at the G2/M phase and induced apoptosis at sub μM concentrations. Moreover, they caused the destruction of a preformed vascular network in vitro and inhibited the migration of endothelial cells at non-toxic concentrations. Finally, these compounds displayed high affinity for tubulin as substantiated by a Kb value of 2.87 × 108 M−1 which, to the best of our knowledge, represents the highest binding constant measured to date for a colchicine-domain ligand. This project has been supported by the Spanish Ministerio de Economía y Competitividad (SAF2015–64629- C2-1-R to EMP, MJPP. and MJC and SAF2015-64629-C2-2 to F.G.), by the Swiss National Science Foundation (31003A_166608, to MOS) and COST action CM1407 (to MJPP, SL, AP, MS, and JFD). We acknowledge Lizette Van Berckelaer, Eef Meyen and Sam Noppen for excellent technical assistance. We thank V. Olieric and M. Wang for excellent technical assistance with the collection of X-ray data at beamline X06SA of the Swiss Light Source (Paul Scherrer Institut, Villigen PSI, Switzerland). We thank José Fernando Escolar for his help with electron microscopy. We thank ganadería Fernando Díaz for calf brains for tubulin purification. This work was supported in part by grants BFU2016-75319-R (AEI/FEDER, UE) (JFD.) JFD. is a member of the CIB Intramural Program “Molecular Machines for Better Life” (MACBET). We also thank SEQT for the “Ramón Madroñero Award” to O.B. |
| Databáze: | OpenAIRE |
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