Interaction of Vinca Alkaloids with Tubulin: A Comparison of Vinblastine, Vincristine, and Vinorelbine †

School of Nursing, University of Mississippi Medical Center, Jackson 39216, USA.
Biochemistry (Impact Factor: 3.02). 06/1996; 35(21):6806-14. DOI: 10.1021/bi953037i
Source: PubMed


Vinca alkaloids are antimitotic drugs that inhibit microtubule assembly and induce tubulin self-association into coiled spiral aggregates. Previous sedimentation velocity results with vinblastine have been interpreted by a mechanism involving isodesmic ligand-mediated or ligand-mediated plus ligand-facilitated self-association [Lobert et al. (1995) Biochemistry 34, 8050-8060]. In this study, we compare the vincristine- or vinorelbine-induced self-association of porcine brain tubulin with our prior vinblastine studies in the presence of 50 microM GDP or 50 microM GTP. Vincristine demonstrates the highest overall affinity for tubulin, K1K2, and vinorelbine the lowest (vincristine > vinblastine > vinorelbine). These and the first quantitative studies comparing the interaction of a new vinca alkaloid derivative, vinorelbine (Navelbine), with other vinca alkaloids. The relative binding affinities reported here correlate with the weekly drug doses used clinically in cancer chemotherapy, where vincristine is used at the lowest dosages and vinorelbine at the highest. Surprisingly, K1, the affinity of drug for tubulin heterodimers, is identical for all three drugs. When data are fit with the ligand-mediated model, the differences in overall affinity are due to effects on K2, the affinity of liganded heterodimers for spiral polymers. When data are fit with the ligand-mediated plus-facilitated model, affinity differences are also reflected in K3, the binding of the drug to unliganded polymers. We find that GDP enhances self-association in the presence of all three drugs 3-5-fold over GTP. The enhancement is manifested in K2 and K3 and amounts to an average of 0.90 +/= 0.17 kcal/mol. Thus, nucleotide enhancement is linked to the self-association step. Data collected at 5, 25, and 36 degrees C for all three drugs show increased maximum s-20,w values with increasing temperature and are consistent with an entropically driven reaction for the overall process. To investigate these results further, stopped-flow light scattering experiments have been conducted. Relaxation times are longest for the largest vincristine polymers and shortest for the smallest vinorelbine polymers, consistent with a cascade of events corresponding to successive dissociation events from spiral polymers, the larger the polymers the longer the relaxation time. Relaxation times for any single drug decrease with increasing tubulin concentration, consistent with the occurrence of oligomer annealing in addition to the association of liganded heterodimers to the ends of the growing spirals. Relaxation times were used to estimate on and off rates for liganded heterodimer association with spirals, and their ratio gives affinity constants (Kapp) that are independently consistent with K2 estimates from sedimentation velocity results for vinblastine and vinorelbine. For vincristine-induced tubulin polymers, a two-step process is observed with a second relaxation time more than 20-fold longer than times observed for vinblastine or vinorelbine. Sedimentation velocity experiments at low speeds and electron microscopy are consistent with the presence of a small amount of larger polymers (> or = 40S) in the vincristine samples, possibly involving alignment of spirals. Under our experimental conditions, these larger polymers appear to have a minimal effect on the estimated energetics of the vincristine-induced self-association of tubulin.

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Available from: John Correia, Aug 24, 2015
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    • "Antitubulin vinca alkaloids prevent tubulin polymerization from soluble dimers into microtubules. The affinity for tubulin differs among vinca-alkaloid compounds (decreasing in order vincristine , vinblastine, vinorelbine), which might explain the distinct neurotoxic profiles of these drugs [37]. Vincristine is the most neurotoxic one, vinblastine, and vinorelbine are less neurotoxic. "
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    • "The incidence and severity of vincristine-induced neuropathic pain is positively correlated with the duration of treatment and doses used. The anti-tumour action of vincristine is due to its high binding affinity to β-tubulin, leading to aborted cell division and cell death (Wilson et al. 1975; Lobert et al. 1996). An animal model of vincristine-induced nociceptive sensory neuropathy was previously developed and behaviourally characterized (Aley et al. 1996; Nozaki-Taguchi et al. 2001; Authier et al. 2003). "
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