Unravelling the covalent binding of zampanolide and taccalonolide AJ to a minimalist representation of a human microtubule

Sánchez‐Murcia, Pedro A.; Mills, Alberto; Cortés-Cabrera, Álvaro; Gago, Federico

doi:10.1007/s10822-019-00208-w

Cited by 13 publications

(27 citation statements)

References 112 publications

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“…L217, R369, L219, L230, L371, Y283, G225, G370, and S277). Overall, the prediction of key interacting residues based on covalent docking to the 5EZY crystallographic structure qualitatively matched the molecular dynamics simulation results of 2 using the cryo-EM structure of a mammalian microtubule 26 . Thus, besides D226, 6 β-tubulin residues (i.e., K19, H229, R278, L217, L219, and T223) were selected for mutagenesis with similar procedures, as described above ( Supplementary Table 9) followed by immunoblotting to determine their impact on taccalonolide binding.…”

Section: Resultssupporting

confidence: 59%

“…Therefore, the opening of the 22,23-epoxy group of 2 is likely facilitated via direct nucleophilic attack by the carboxylate of βtubulin D226 (Fig. 1d) 26 . This epoxide opening mechanism was supported by covalent docking of 2 into β-tubulin using CovDock affording a lowest-energy docking model that perfectly matched the 5EZY crystal structure (RMSD = 0.221, Fig.…”

Section: Resultsmentioning

confidence: 99%

“…1e and 7a). The T223 hydroxyl was also predicted to play an important role in fixing the carboxylate of D226 to facilitate the covalent reaction with the 22,23-epoxide on the basis of the molecular dynamics simulation of 2 26 . Interestingly, the T223A mutation only had a moderate effect on binding (Fig.…”

Section: Resultsmentioning

confidence: 99%

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Elucidating target specificity of the taccalonolide covalent microtubule stabilizers employing a combinatorial chemical approach

Yee

Ramachandran

et al. 2020

Nat Commun

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The taccalonolide microtubule stabilizers covalently bind β-tubulin and overcome clinically relevant taxane resistance mechanisms. Evaluations of the target specificity and detailed drug-target interactions of taccalonolides, however, have been limited in part by their irreversible target engagement. In this study, we report the synthesis of fluorogenic taccalonolide probes that maintain the native biological properties of the potent taccalonolide, AJ. These carefully optimized, cell-permeable probes outperform commercial taxane-based probes and enable direct visualization of taccalonolides in both live and fixed cells with dramatic microtubule colocalization. The specificity of taccalonolide binding to β-tubulin is demonstrated by immunoblotting, which allows for determination of the relative contribution of key tubulin residues and taccalonolide moieties for drug-target interactions by activity-based protein profiling utilizing site-directed mutagenesis and computational modeling. This combinatorial approach provides a generally applicable strategy for investigating the binding specificity and molecular interactions of covalent binding drugs in a cellular environment.

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Section: Resultssupporting

confidence: 59%

Section: Resultsmentioning

confidence: 99%

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Elucidating target specificity of the taccalonolide covalent microtubule stabilizers employing a combinatorial chemical approach

Yee

Ramachandran

et al. 2020

Nat Commun

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“…Taccalonolide AF marked a milestone in understanding the structure-activity relationships (SARs) of taccalonolides and the mechanism of action of this class of compounds. The absolute configuration of the C22–C23 epoxide of taccalonolides AF and AJ was determined to be R , R , by single-crystal X-ray diffraction analysis [ 17 , 18 ].…”

Section: Structures Of Natural Taccalonolides (1987–2020)mentioning

confidence: 99%

“…These SARs are observed as well by computational analysis such as i) taccalonolides covalently bind to β-tubulin D226 via the C22–C23 epoxide group, and ii) the bulky group at C-1 is involved in interactions with β-tubulin residues through hydrogen bonds [ 17 , 18 , 45 ]. According to established SARs, a group of taccalonolides with a fluorescein group at C-6 such as Flu-tacca-4, Flu-tacca-5, Flu-tacca-7, and Flu-tacca-8 were designed and generated to facilitate the identification of binding site interactions, among which Flu-tacca-7 showed comparable potencies in the proliferation of HeLa and SK-OV-3 cells ( Figure 6 ) [ 18 , 46 ].…”

Section: Semisynthetic Taccalonolides and Structure-activity Relatmentioning

confidence: 99%

Taccalonolides: A Novel Class of Microtubule-Stabilizing Anticancer Agents

Chen

Winstead

et al. 2021

Cancers

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Microtubule stabilizing agents, such as paclitaxel, docetaxel, and cabazitaxel have been among the most used chemotherapeutic agents in the last decades for the treatment of a wide range of cancers in the clinic. One of the concerns that limit their use in clinical practice is their intrinsic and acquired drug resistance, which is common to most anti-cancer chemotherapeutics. Taccalonolides are a new class of microtubule stabilizers isolated from the roots of a few species in the genus of Tacca. In early studies, taccalonolides demonstrated different effects on interphase and mitotic microtubules from those of paclitaxel and laulimalide suggesting a unique mechanism of action. This prompts the exploration of new taccalonolides with various functionalities through the identification of minor constituents of natural origin and semi-synthesis. The experiments on the new highly potent taccalonolides indicated that taccalonolides possessed a unique mechanism of covalently binding to the microtubule. An X-ray diffraction analysis of a crystal of taccalonolides AJ binding to tubulin indicated that the covalent binding site is at β-tubulin D226. Taccalonolides circumvent all three mechanisms of taxane drug resistance both in vitro and in vivo. To improve the activity, the structure modification through semi-synthesis was conducted and the structure-activity relationships (SARs) was analyzed based on natural and semi-synthetical taccalonolides. The C22-C23 epoxide can significantly increase the antiproliferation potency of taccalonolides due to the covalent link of C22 and the carboxylic group of D226. Great progress has been seen in the last few years in the understanding of the mechanism of this class of microtube-stabilizing agents. This review summarizes the structure diversity, structure-activity relationships (SARs), mechanism of action, and in vivo activities of taccalonolides.

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Microtubule‐targeting agents for cancer treatment: Seven binding sites and three strategies

Wang,

Gigant,

Zheng

et al. 2023

MedComm – Oncology

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Microtubules are pivotal in diverse cellular functions encompassing cell signaling, morphology, intracellular trafficking, and cell mitosis/division. They are validated targets for disease treatment, notably hematological cancers and solid tumors. Microtubule‐targeting agents (MTAs) exert their effects by modulating microtubule dynamics, impeding cell proliferation, and promoting cell death. Recent advances in structural biology have unveiled novel perspectives for investigating multiple binding sites and mechanisms of action used by MTAs. In this review, we first provide an overview of the intricate structure and dynamics of microtubules. Then we explore the seven binding sites and the three primary strategies (stabilization, destabilization, and degradation) harnessed by MTAs. Furthermore, we introduce the emerging domain of microtubule‐targeting degraders, exemplified by PROteolysis TArgeting Chimeras and small‐molecule degraders, which enable precise degradation of specific microtubule‐associated proteins implicated in cancer pathogenesis. Additionally, we discuss the promising realm of precision‐targeted approaches, including antibody–drug conjugates and the utilization of photopharmacology in MTAs. Lastly, we provide a comprehensive overview of the clinical applications of microtubule‐targeting therapies, assessing their efficacy and current challenges. We aim to provide a global picture of MTAs development as well as insights into the microtubule‐targeting drug discovery for cancer treatment.

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Unravelling the covalent binding of zampanolide and taccalonolide AJ to a minimalist representation of a human microtubule

Cited by 13 publications

References 112 publications

Elucidating target specificity of the taccalonolide covalent microtubule stabilizers employing a combinatorial chemical approach

Elucidating target specificity of the taccalonolide covalent microtubule stabilizers employing a combinatorial chemical approach

Taccalonolides: A Novel Class of Microtubule-Stabilizing Anticancer Agents

Microtubule‐targeting agents for cancer treatment: Seven binding sites and three strategies

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