title: Antimitotic and tubulin-interacting properties of vinflunine, a novel fluorinated Vinca alkaloid.
abstract: This study aimed to define the mechanism of action of vinflunine, a novel Vinca alkaloid synthesised from vinorelbine using superacidic chemistry and characterised by superior in vivo activity to vinorelbine in preclinical tumour models. In vitro vinflunine cytotoxicity proved dependent on concentration and exposure duration, with IC50 values (72-hr exposures) generally ranging from 60-300 nM. Vinflunine induced G2 + M arrest, associated with mitotic accumulation and a concentration-dependent reduction of the microtubular network of interphase cells, accompanied by paracrystal formation. These effects, while comparable to those of vincristine, vinblastine or vinorelbine, were achieved with 3- to 17-fold higher vinflunine concentrations. However, vinflunine and the other Vincas all inhibited microtubule assembly at micromolar concentrations. Vinflunine, like vinblastine, vincristine and vinorelbine, appeared to interact at the Vinca binding domain, as judged by proteolytic cleavage patterns, and induced tubulin structural changes favouring an inhibition of GTP hydrolysis. However, vinflunine did not prevent [3H]vincristine binding to unassembled tubulin at concentrations < or = 100 microM, and only weakly inhibited binding of [3H]vinblastine or [3H]vinorelbine. Indeed, specific binding of [3H]vinflunine to tubulin was undetectable by centrifugal gel filtration. Thus, the comparative capacities of these Vincas to bind to or to interfere with their binding to tubulin could be classified as: vincristine > vinblastine > vinorelbine > vinflunine. By monitoring alkylation of sulfhydryl groups, differential effects on tubulin conformation were identified with vinflunine and vinorelbine acting similarly, yet distinctively from vinblastine and vincristine. Overall, vinflunine appears to function as a definite inhibitor of tubulin assembly, while exhibiting quantitatively different tubulin binding properties to the classic Vinca alkaloids.
authors.affiliation: Division of Experimental Cancer Research I, Centre de Recherche Pierre Fabre, Castres, France.
authors.forename: A
authors.initials: A
authors.lastname: Kruczynski
authors.forename: J M
authors.initials: JM
authors.lastname: Barret
authors.forename: C
authors.initials: C
authors.lastname: Etiévant
authors.forename: F
authors.initials: F
authors.lastname: Colpaert
authors.forename: J
authors.initials: J
authors.lastname: Fahy
authors.forename: B T
authors.initials: BT
authors.lastname: Hill
mesh_terms: D000478:Alkylation; D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D002453:Cell Cycle / Q000187:drug effects; D002478:Cells, Cultured; D006160:Guanosine Triphosphate / Q000378:metabolism; D006868:Hydrolysis; D007460:Iodoacetamide / Q000494:pharmacology; D007939:Leukemia L1210 / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008938:Mitosis / Q000187:drug effects*; D011485:Protein Binding; D051381:Rats; D014404:Tubulin / Q000187:drug effects* / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014747:Vinblastine / Q000031:analogs & derivatives* / Q000494:pharmacology; D014750:Vincristine / Q000494:pharmacology; D000077235:Vinorelbine
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D014404:Tubulin; C111217:vinflunine; D014750:Vincristine; D014747:Vinblastine; D006160:Guanosine Triphosphate; D000077235:Vinorelbine; D007460:Iodoacetamide