Biarylacetamides: a novel class of late-stage autophagy inhibitors

Targeting autophagy is believed to hold great promise for the treatment of various diseases, including cancer. However, since the therapeutic efficacy of currently available autophagy-modulating drugs is limited by off-target effects and the requirement of high dosage, there is an urgent need to dev...

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Main Authors: Mélissa Lallier, Rani Robeyns, Freke Mertens, Angela Sisto, Guido R.Y. De Meyer, Koen Augustyns, Maya Berg, Winnok H. De Vos, Vincent Timmerman, George M.C. Janssen, Peter van Veelen, Alexander L.N. van Nuijs, Nikolai Engedal, Wim Martinet, Pieter Van der Veken
Format: Article
Language:English
Published: Taylor & Francis Group 2025-12-01
Series:Autophagy Reports
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Online Access:https://www.tandfonline.com/doi/10.1080/27694127.2025.2541597
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author Mélissa Lallier
Rani Robeyns
Freke Mertens
Angela Sisto
Guido R.Y. De Meyer
Koen Augustyns
Maya Berg
Winnok H. De Vos
Vincent Timmerman
George M.C. Janssen
Peter van Veelen
Alexander L.N. van Nuijs
Nikolai Engedal
Wim Martinet
Pieter Van der Veken
author_facet Mélissa Lallier
Rani Robeyns
Freke Mertens
Angela Sisto
Guido R.Y. De Meyer
Koen Augustyns
Maya Berg
Winnok H. De Vos
Vincent Timmerman
George M.C. Janssen
Peter van Veelen
Alexander L.N. van Nuijs
Nikolai Engedal
Wim Martinet
Pieter Van der Veken
author_sort Mélissa Lallier
collection DOAJ
description Targeting autophagy is believed to hold great promise for the treatment of various diseases, including cancer. However, since the therapeutic efficacy of currently available autophagy-modulating drugs is limited by off-target effects and the requirement of high dosage, there is an urgent need to develop novel autophagy-targeting compounds. In this study, we report molecules of the biarylacetamide class as novel autophagy inhibitors. These molecules were identified via phenotypic high-throughput screening, and a series of analogues was subsequently synthesized. Among these, 5d and 5j were retained as potent autophagy blockers in HeLa and LNCaP cells. Both compounds inhibited autophagy at a late-stage in the pathway, as evidenced by the strong accumulation of RFP-GFP-LC3 puncta as well as LC3-II, GABARAP-II and SQSTM1 protein levels, resembling the effects obtained with the well-known late-stage autophagy inhibitor Bafilomycin A1. Quantitative proteome profiling combined with metabolomic and lipidomic studies revealed that 5j significantly altered lipid metabolism. These alterations included activation of the cholesterol biosynthesis pathway and changes in the distribution of key lipid classes, such as phospholipids, ceramides and triglycerides. Further mechanistic studies indicated that 5d and 5j triggered an ER stress response and may impair lysosomal function, as suggested by the accumulation of pro-cathepsin D. Collectively, these findings demonstrate that 5j is a novel and potent late-stage autophagy inhibitor with a distinct mechanism of action compared to currently available inhibitors.
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spelling doaj-art-4049817c772f45bfbe624c4dac8b66352025-08-20T03:41:57ZengTaylor & Francis GroupAutophagy Reports2769-41272025-12-014110.1080/27694127.2025.2541597Biarylacetamides: a novel class of late-stage autophagy inhibitorsMélissa Lallier0Rani Robeyns1Freke Mertens2Angela Sisto3Guido R.Y. De Meyer4Koen Augustyns5Maya Berg6Winnok H. De Vos7Vincent Timmerman8George M.C. Janssen9Peter van Veelen10Alexander L.N. van Nuijs11Nikolai Engedal12Wim Martinet13Pieter Van der Veken14Laboratory of Medicinal Chemistry, Department of Pharmaceutical Sciences, University of Antwerp, Antwerp, BelgiumToxicological Centre, Department of Pharmaceutical Sciences, University of Antwerp, Antwerp, BelgiumLaboratory of Physiopharmacology, Department of Pharmaceutical Sciences, University of Antwerp, Antwerp, BelgiumPeripheral Neuropathy Research Group, Department of Biomedical Sciences, University of Antwerp, Antwerpen, BelgiumLaboratory of Physiopharmacology, Department of Pharmaceutical Sciences, University of Antwerp, Antwerp, BelgiumLaboratory of Medicinal Chemistry, Department of Pharmaceutical Sciences, University of Antwerp, Antwerp, BelgiumInfla-Med Centre of Excellence, University of Antwerp, Antwerpen, BelgiumAntwerp Centre for Advanced Microscopy, University of Antwerp, Antwerp, BelgiumPeripheral Neuropathy Research Group, Department of Biomedical Sciences, University of Antwerp, Antwerpen, BelgiumCenter for Proteomics and Metabolomics, Leiden University Medical Center (LUMC), Leiden, NetherlandsCenter for Proteomics and Metabolomics, Leiden University Medical Center (LUMC), Leiden, NetherlandsToxicological Centre, Department of Pharmaceutical Sciences, University of Antwerp, Antwerp, BelgiumAutophagy in Cancer Lab, Department of Tumor Biology, Oslo University Hospital, Oslo, NorwayLaboratory of Physiopharmacology, Department of Pharmaceutical Sciences, University of Antwerp, Antwerp, BelgiumLaboratory of Medicinal Chemistry, Department of Pharmaceutical Sciences, University of Antwerp, Antwerp, BelgiumTargeting autophagy is believed to hold great promise for the treatment of various diseases, including cancer. However, since the therapeutic efficacy of currently available autophagy-modulating drugs is limited by off-target effects and the requirement of high dosage, there is an urgent need to develop novel autophagy-targeting compounds. In this study, we report molecules of the biarylacetamide class as novel autophagy inhibitors. These molecules were identified via phenotypic high-throughput screening, and a series of analogues was subsequently synthesized. Among these, 5d and 5j were retained as potent autophagy blockers in HeLa and LNCaP cells. Both compounds inhibited autophagy at a late-stage in the pathway, as evidenced by the strong accumulation of RFP-GFP-LC3 puncta as well as LC3-II, GABARAP-II and SQSTM1 protein levels, resembling the effects obtained with the well-known late-stage autophagy inhibitor Bafilomycin A1. Quantitative proteome profiling combined with metabolomic and lipidomic studies revealed that 5j significantly altered lipid metabolism. These alterations included activation of the cholesterol biosynthesis pathway and changes in the distribution of key lipid classes, such as phospholipids, ceramides and triglycerides. Further mechanistic studies indicated that 5d and 5j triggered an ER stress response and may impair lysosomal function, as suggested by the accumulation of pro-cathepsin D. Collectively, these findings demonstrate that 5j is a novel and potent late-stage autophagy inhibitor with a distinct mechanism of action compared to currently available inhibitors.https://www.tandfonline.com/doi/10.1080/27694127.2025.2541597Autophagy inhibitorbiarylacetamide derivativesproteomicsmetabolomicslipidomicscholesterol biosynthesis
spellingShingle Mélissa Lallier
Rani Robeyns
Freke Mertens
Angela Sisto
Guido R.Y. De Meyer
Koen Augustyns
Maya Berg
Winnok H. De Vos
Vincent Timmerman
George M.C. Janssen
Peter van Veelen
Alexander L.N. van Nuijs
Nikolai Engedal
Wim Martinet
Pieter Van der Veken
Biarylacetamides: a novel class of late-stage autophagy inhibitors
Autophagy Reports
Autophagy inhibitor
biarylacetamide derivatives
proteomics
metabolomics
lipidomics
cholesterol biosynthesis
title Biarylacetamides: a novel class of late-stage autophagy inhibitors
title_full Biarylacetamides: a novel class of late-stage autophagy inhibitors
title_fullStr Biarylacetamides: a novel class of late-stage autophagy inhibitors
title_full_unstemmed Biarylacetamides: a novel class of late-stage autophagy inhibitors
title_short Biarylacetamides: a novel class of late-stage autophagy inhibitors
title_sort biarylacetamides a novel class of late stage autophagy inhibitors
topic Autophagy inhibitor
biarylacetamide derivatives
proteomics
metabolomics
lipidomics
cholesterol biosynthesis
url https://www.tandfonline.com/doi/10.1080/27694127.2025.2541597
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