Campiani Giuseppe, Khan Tuhina, Ulivieri Cristina, Staiano Leopoldo, Papulino Chiara, Magnano Stefania, Nathwani Seema, Ramunno Anna, Lucena-Agell Daniel, Relitti Nicola, Federico Stefano, Pozzetti Luca, Carullo Gabriele, Casagni Alice, Brogi Simone, Vanni Francesca, Galatello Paola, Ghanim Magda, McCabe Niamh, Lamponi Stefania, Valoti Massimo, Ibrahim Ola, O'Sullivan Jeffrey, Turkington Richard, Kelly Vincent P, VanWemmel Ruben, Díaz J Fernando, Gemma Sandra, Zisterer Daniela, Altucci Lucia, De Matteis Antonella, Butini Stefania, Benedetti Rosaria
Department of Biotechnology, Chemistry and Pharmacy, DoE Department of Excellence 2018-2022, University of Siena, via Aldo Moro 2, 53100, Siena, Italy.
Department of Biotechnology, Chemistry and Pharmacy, DoE Department of Excellence 2018-2022, University of Siena, via Aldo Moro 2, 53100, Siena, Italy.
Eur J Med Chem. 2022 May 5;235:114274. doi: 10.1016/j.ejmech.2022.114274. Epub 2022 Mar 20.
Autophagy is a lysosome dependent cell survival mechanism and is central to the maintenance of organismal homeostasis in both physiological and pathological situations. Targeting autophagy in cancer therapy attracted considerable attention in the past as stress-induced autophagy has been demonstrated to contribute to both drug resistance and malignant progression and recently interest in this area has re-emerged. Unlocking the therapeutic potential of autophagy modulation could be a valuable strategy for designing innovative tools for cancer treatment. Microtubule-targeting agents (MTAs) are some of the most successful anti-cancer drugs used in the clinic to date. Scaling up our efforts to develop new anti-cancer agents, we rationally designed multifunctional agents 5a-l with improved potency and safety that combine tubulin depolymerising efficacy with autophagic flux inhibitory activity. Through a combination of computational, biological, biochemical, pharmacokinetic-safety, metabolic studies and SAR analyses we identified the hits 5i,k. These MTAs were characterised as potent pro-apoptotic agents and also demonstrated autophagy inhibition efficacy. To measure their efficacy at inhibiting autophagy, we investigated their effects on basal and starvation-mediated autophagic flux by quantifying the expression of LC3II/LC3I and p62 proteins in oral squamous cell carcinoma and human leukaemia through western blotting and by immunofluorescence study of LC3 and LAMP1 in a cervical carcinoma cell line. Analogues 5i and 5k, endowed with pro-apoptotic activity on a range of hematological cancer cells (including ex-vivo chronic lymphocytic leukaemia (CLL) cells) and several solid tumor cell lines, also behaved as late-stage autophagy inhibitors by impairing autophagosome-lysosome fusion.
自噬是一种依赖溶酶体的细胞存活机制，在生理和病理情况下对于维持机体稳态至关重要。过去，由于应激诱导的自噬已被证明与耐药性和恶性进展均有关，因此在癌症治疗中靶向自噬备受关注，并且最近该领域的研究兴趣再度兴起。释放自噬调节的治疗潜力可能是设计创新癌症治疗工具的一项有价值策略。微管靶向剂（MTAs）是迄今为止临床上使用的一些最成功的抗癌药物。为加大开发新型抗癌药物的力度，我们合理设计了多功能试剂5a-l，其效力和安全性均有所提高，将微管蛋白解聚功效与自噬流抑制活性相结合。通过计算、生物学、生物化学、药代动力学-安全性、代谢研究及构效关系分析，我们确定了活性化合物5i、k。这些MTAs被表征为强效促凋亡剂，同时也显示出自噬抑制功效。为测定它们抑制自噬的效力，我们通过蛋白质免疫印迹法量化口腔鳞状细胞癌和人白血病中LC3II/LC3I和p62蛋白的表达，并通过对宫颈癌细胞系中的LC3和LAMP1进行免疫荧光研究，调查它们对基础和饥饿介导的自噬流的影响。类似物5i和5k对一系列血液癌细胞（包括体外慢性淋巴细胞白血病（CLL）细胞）和几种实体瘤细胞系具有促凋亡活性，还通过损害自噬体-溶酶体融合而表现为晚期自噬抑制剂。
