Synthesis and Superpotent Anticancer Activity of Tubulysins Carrying Non-hydrolysable N-Substituents on Tubuvaline

Monica Sani, Paolo Lazzari, Marco Folini, Marco Spiga, Valentina Zuco, Michelandrea De Cesare, Ilaria Manca, Sergio Dall'Angelo, Massimo Frigerio, Igor Usai, Andrea Testa, Nadia Zaffaroni, Matteo Zanda

Research output: Contribution to journalArticlepeer-review


Synthetic tubulysins 24 a–m, containing non-hydrolysable N-substituents on tubuvaline (Tuv), were obtained in high purity and good overall yields using a multistep synthesis. A key step was the formation of differently N-substituted Ile-Tuv fragments 10 by using an aza-Michael reaction of azido-Ile derivatives 8 with the α,β-unsaturated oxo-thiazole 5. A structure–activity relationship study using a panel of human tumour cell lines showed strong anti-proliferative activity for all compounds 24 a–m, with IC50 values in the sub-nanomolar range, which were distinctly lower than those of tubulysin A, vinorelbine and paclitaxel. Furthermore, 24 a–m were able to overcome cross-resistance to paclitaxel and vinorelbine in two tumour cell lines with acquired resistance to doxorubicin. Compounds 24 e and 24 g were selected as leads to evaluate their mechanism of action. In vitro assays showed that both 24 e and 24 g interfere with tubulin polymerization in a vinca alkaloid-like manner and prevent paclitaxel-induced assembly of tubulin polymers. Both compounds exerted antimitotic activity and induced apoptosis in cancer cells at very low concentrations. Compound 24 e also exhibited potent antitumor activity at well tolerated doses on in vivo models of diffuse malignant peritoneal mesothelioma, such as MESOII peritoneal mesothelioma xenografts, the growth of which was not significantly affected by vinorelbine. These results indicate that synthetic tubulysins 24 could be used as standalone chemotherapeutic agents in difficult-to-treat cancers.

Original languageEnglish
Pages (from-to)5842-5850
Number of pages9
JournalChemistry - A European Journal
Issue number24
Publication statusPublished - Apr 27 2017


  • antitumor agents
  • Michael addition
  • peptides
  • structure–activity relationships
  • tubulysins

ASJC Scopus subject areas

  • Chemistry(all)


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