Tubulin-Binding Drug Screening by MALDI-TOFMS

Abstract

Despite a large amount of drugs available to treat cancer, none is totally satisfactory with respect to its tolerance or side effects. It is very important to discover new compounds that exhibit specific features such as binding to proteic targets. Given the clinical successes of the poisons of the mitotic spindle chemotherapeutic agent class, it is often considered that tubulin represents one of the best cancer targets identified so far, and it seems likely that discovering new drugs of this class will significantly improve the range of active chemotherapeutic agents. The aim of this work is to present the new screening test that has been developed in our laboratory in order to study the binding of compounds to tubulin. We have developed a screening protocol involving three sampling strategies before the MALDI-TOFMS analysis. The three strategies give very accurate and reproducible results and could therefore possibly be used in screening campaigns. We have also proved that no unspecific binding can provide a loss of specificity of the test. Our protocol presents all the requirements for being a useful tool to screen the binding of compounds to tubulin.

Mass spectrometric study of colchicine and its synthetic derivatives 10-alkylthiocolchicines

“…24,25 This fact has prompted us to examine the series of other biologically active colchicine derivatives, i.e.…”

Screening and Determination for Potential α‐Glucosidase Inhibitors from Leaves of Acanthopanax senticosus Harms by Using UF‐LC/MS and ESI‐MSn

“…To overcome the limitations of the in vitro screening assays and enhance the throughput of drug discovery, mass spectrometry (MS)‐based techniques have been widely applied in recent years, including size‐exclusion chromatography LC/MS (Wabnitz and Loo, ), ultrafiltration (UF)‐LC/MS (Liu et al ., ), frontal affinity chromatography LC/MS (Ng et al ., ), capillary MS (Hodgson et al ., ), surface plasmon resonance (SPR)‐MS (Marchesini et al ., ), matrix‐assisted laser desorption/ionisation (MALDI) time of flight (TOF) MS (Hannewald et al ., ), and quantitative MALDI‐Fourier transform (FT) MS (Xu et al ., ). UF‐LC/MS has been proven to be a powerful tool for screening biologically active compounds from botanical extracts because the ultrafiltration step facilitates the separation of ligand‐receptor complexes from unbound compounds, followed by LC/MS identification of the ligands.…”

Diagnosis of Biological Activities by Mass Spectrometry

“…The complex protein-ligand is analyzed by Mass Spectrometry and the direct detection of the ligand reflects its potential inhibition activity against the target. Screening tests have already been developed for the ligands of tubulin [2,3], dihydroxyfolate reductase (DHFR) [4] and recently for reversible and irreversible ligands of CDC25 phosphatases (isoforms A et C) [5]. This approach demonstrates its effectiveness with Madagascar periwinkle for tubulin, with Colchicum and green tea for DHFR and some promising plants are currently evaluated on CDC25s.…”