title: New chemical tools for investigating human mitotic kinesin Eg5.
abstract: We have designed and synthesized a series of monastrol derivatives, an allosteric inhibitor of Eg5, a motor protein responsible for the formation and maintenance of the bipolar spindle in mitotic cells. Sterically demanding structural modifications have been introduced on the skeleton of the parent drug either via a multicomponent Biginelli reaction or a stepwise modification of monastrol. The ability of these compounds to inhibit Eg5 activity has been investigated using two in vitro steady-state ATPase assays (basal and microtubule-stimulated) as well as a cell-based assay. One compound in the series appeared more potent than monastrol by a fivefold factor. Three other compounds that were unable to inhibit Eg5 ATPase activity in vitro proved potent Eg5 inhibitors in the cell-based assay. The results obtained led to the identification of structure-activity relationships further used to design an affinity matrix that can be used for fast and efficient purification of Eg5 from crude lysate of eukaryotic cells.
authors.affiliation: Institut Gilbert-Laustriat, CNRS-Université Louis Pasteur, Faculté de Pharmacie, 74 route du Rhin, 67401 Illkirch, France.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Klein
authors.forename: Salvatore
authors.initials: S
authors.lastname: DeBonis
authors.forename: Bernd
authors.initials: B
authors.lastname: Thiede
authors.forename: Dimitrios A
authors.initials: DA
authors.lastname: Skoufias
authors.forename: Frank
authors.initials: F
authors.lastname: Kozielski
authors.forename: Luc
authors.initials: L
authors.lastname: Lebeau
mesh_terms: D050256:Antimitotic Agents / Q000138:chemical synthesis / Q000494:pharmacology*; D001681:Biological Assay; D002478:Cells, Cultured; D015195:Drug Design; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000494:pharmacology*; D005057:Eukaryotic Cells; D006801:Humans; D016547:Kinesins / Q000037:antagonists & inhibitors*; D008870:Microtubules / Q000378:metabolism; D008938:Mitosis / Q000187:drug effects*; D011743:Pyrimidines / Q000138:chemical synthesis / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D013871:Thiones / Q000138:chemical synthesis / Q000494:pharmacology*
chemical_list: D050256:Antimitotic Agents; D004791:Enzyme Inhibitors; C470243:KIF11 protein, human; D011743:Pyrimidines; D013871:Thiones; C400223:monastrol; D016547:Kinesins