Design and synthesis of multifunctional microtubule targeting agents endowed with dual pro-apoptotic and anti-autophagic efficacy PMID: 35344902

ADC Targets in Oncology

2026-04-10

Design and synthesis of multifunctional microtubule targeting agents endowed with dual pro-apoptotic and anti-autophagic efficacy

Eur J Med Chem. 2022 May 5:235:114274. doi: 10.1016/j.ejmech.2022.114274.

Giuseppe Campiani ¹ , Tuhina Khan ² , Cristina Ulivieri ³ , Leopoldo Staiano ⁴ , Chiara Papulino ⁵ , Stefania Magnano ⁶ , Seema Nathwani ⁶ , Anna Ramunno ⁷ , Daniel Lucena-Agell ⁸ , Nicola Relitti ⁹ , Stefano Federico ² , Luca Pozzetti ² , Gabriele Carullo ² , Alice Casagni ² , Simone Brogi ¹⁰ , Francesca Vanni ¹¹ , Paola Galatello ⁷ , Magda Ghanim ⁶ , Niamh McCabe ¹² , Stefania Lamponi ² , Massimo Valoti ¹¹ , Ola Ibrahim ¹³ , Jeffrey O'Sullivan ¹³ , Richard Turkington ¹² , Vincent P Kelly ⁶ , Ruben VanWemmel ⁸ , J Fernando Díaz ⁸ , Sandra Gemma ² , Daniela Zisterer ⁶ , Lucia Altucci ¹⁴ , Antonella De Matteis ¹⁵ , Stefania Butini ¹⁶ , Rosaria Benedetti ⁵

Affiliations

1. Department of Biotechnology, Chemistry and Pharmacy, DoE Department of Excellence 2018-2022, University of Siena, via Aldo Moro 2, 53100, Siena, Italy. Electronic address: [email protected].
2. Department of Biotechnology, Chemistry and Pharmacy, DoE Department of Excellence 2018-2022, University of Siena, via Aldo Moro 2, 53100, Siena, Italy.
3. Department of Life Sciences, University of Siena, via Aldo Moro 2, 53100, Siena, Italy; Istituto Toscano Tumori, University of Siena, via Aldo Moro 2, I, 53100, Siena, Italy.
4. Cell Biology and Disease Mechanisms, Telethon Institute of Genetics and Medicine, Via Campi Flegrei, 34, 80078, Pozzuoli, Naples, Italy; Institute for Genetic and Biomedical Research, National Research Council (CNR), via Fratelli Cervi 93, 20054, Segrate, Milan, Italy.
5. Department of Precision Medicine, University of Campania Luigi Vanvitelli, Vico L, De Crecchio 7, 80138, Naples, IT, Italy.
6. School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute, Trinity College Dublin, 152-160, Pearse Street, Dublin 2, Ireland.
7. Department of Pharmacy, University of Salerno, via G. Paolo II 132, 84084, Fisciano (SA), Italy.
8. Centro de Investigaciones Biologicas Margarita Salas, Consejo Superior de Investigaciones Cientificas, Ramiro de Maeztu 9, 28040, Madrid, Spain.
9. IRBM Science Park, Via Pontina km 30, 600, 00071, Pomezia, Rome, Italy.
10. Department of Pharmacy, University of Pisa, 56126, Pisa, Italy.
11. Department of Life Sciences, University of Siena, via Aldo Moro 2, 53100, Siena, Italy.
12. School of Medicine, Dentistry and Biomedical Sciences, Queen's University Belfast, 97 Lisburn Road, Health Sciences Building, BT9 7BL, Belfast, United Kingdom.
13. School of Dental Science, Trinity College Dublin, Lincoln Place, Dublin 2, Ireland.
14. Department of Precision Medicine, University of Campania Luigi Vanvitelli, Vico L, De Crecchio 7, 80138, Naples, IT, Italy; Biogem Institute of Molecular Biology and Genetics, Via Camporeale, 83031, Ariano Irpino, Italy.
15. Cell Biology and Disease Mechanisms, Telethon Institute of Genetics and Medicine, Via Campi Flegrei, 34, 80078, Pozzuoli, Naples, Italy.
16. Department of Biotechnology, Chemistry and Pharmacy, DoE Department of Excellence 2018-2022, University of Siena, via Aldo Moro 2, 53100, Siena, Italy; Istituto Toscano Tumori, University of Siena, via Aldo Moro 2, I, 53100, Siena, Italy. Electronic address: [email protected].

Abstract

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.

Keywords

Apoptosis; Autophagy; Autophagy inhibitors; Cancer; Ex vivo; Microtubule-targeting agents.