Reversing TGF-β1-induced fibrotic phenotype in human lung fibroblasts using a PROTAC tool derived from an indoline-based HDAC6 inhibitor

Idiopathic pulmonary fibrosis (IPF) is a progressive and debilitating interstitial lung disease characterized by limited therapeutic options, with only two FDA-approved palliative agents currently available. Given its poor prognosis and the high incidence of lung transplantation, there is a pressing...

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Main Authors: Ilaria Cursaro, Valerio Ciccone, Valeria Tudino, Chiara Papulino, Laura Scalvini, Sandra Donnini, Lucia Morbidelli, Luca Bini, Claudia Landi, Chiara Contri, Silvia Pasquini, Fabrizio Vincenzi, Katia Varani, Alessio Lodola, Marco Mor, Rosaria Benedetti, Lucia Altucci, Stefania Butini, Gabriele Carullo, Sandra Gemma, Giuseppe Campiani
Format: Article
Language:English
Published: Elsevier 2025-08-01
Series:European Journal of Medicinal Chemistry Reports
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Online Access:http://www.sciencedirect.com/science/article/pii/S2772417425000329
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author Ilaria Cursaro
Valerio Ciccone
Valeria Tudino
Chiara Papulino
Laura Scalvini
Sandra Donnini
Lucia Morbidelli
Luca Bini
Claudia Landi
Chiara Contri
Silvia Pasquini
Fabrizio Vincenzi
Katia Varani
Alessio Lodola
Marco Mor
Rosaria Benedetti
Lucia Altucci
Stefania Butini
Gabriele Carullo
Sandra Gemma
Giuseppe Campiani
author_facet Ilaria Cursaro
Valerio Ciccone
Valeria Tudino
Chiara Papulino
Laura Scalvini
Sandra Donnini
Lucia Morbidelli
Luca Bini
Claudia Landi
Chiara Contri
Silvia Pasquini
Fabrizio Vincenzi
Katia Varani
Alessio Lodola
Marco Mor
Rosaria Benedetti
Lucia Altucci
Stefania Butini
Gabriele Carullo
Sandra Gemma
Giuseppe Campiani
author_sort Ilaria Cursaro
collection DOAJ
description Idiopathic pulmonary fibrosis (IPF) is a progressive and debilitating interstitial lung disease characterized by limited therapeutic options, with only two FDA-approved palliative agents currently available. Given its poor prognosis and the high incidence of lung transplantation, there is a pressing need to develop innovative and effective therapeutics. Histone deacetylase 6 (HDAC6) has been identified as a key driver of fibrotic progression in IPF, and selective inhibitors of this isoform were able to revert the fibrotic phenotypes. Proteolysis targeting chimeras (PROTACs) are heterobifunctional molecules that can trigger the degradation of a specific target in cells, including HDAC6 enzyme. Therefore, the application of PROTAC technology may represent a novel therapeutic strategy for IPF. We showcase the design, synthesis, and biological evaluation of a library of first-in-class antifibrotic HDAC6-targeting PROTACs, incorporating our in-house inhibitor 1 as the HDAC6 ligand. Newly developed PROTACs 5a and 5c showed effective degradation of HDAC6 in A549 lung cells and IMR-90 lung fibroblasts. 5a and 5c exhibited significant antifibrotic effects against the TGF-β1 induced fibrotic phenotype on IMR-90 cells, a model that mimics IPF conditions. The generation of putative ternary complexes involving PROTAC molecules, the E3-ligase cereblon (CRBN) and the HDAC6 target protein was supported by molecular modelling techniques, including protein-protein docking and molecular dynamics simulations. Mechanistic investigations, based on pull-down experiments, confirmed that the newly synthetized compounds were able to reduce HDAC6 levels through a proteasome- and CRBN-dependent mechanism as confirmed by experiments with neddylation and proteasome inhibitors. This pioneering exploration of targeted protein degradation in IPF-like conditions provides compelling evidence of its therapeutic promise, paving the way for a broader application of PROTACs in treating rare diseases.
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spelling doaj-art-da891a0adb5446c5bb2fe313fa4ba6a12025-08-20T02:29:19ZengElsevierEuropean Journal of Medicinal Chemistry Reports2772-41742025-08-011410027610.1016/j.ejmcr.2025.100276Reversing TGF-β1-induced fibrotic phenotype in human lung fibroblasts using a PROTAC tool derived from an indoline-based HDAC6 inhibitorIlaria Cursaro0Valerio Ciccone1Valeria Tudino2Chiara Papulino3Laura Scalvini4Sandra Donnini5Lucia Morbidelli6Luca Bini7Claudia Landi8Chiara Contri9Silvia Pasquini10Fabrizio Vincenzi11Katia Varani12Alessio Lodola13Marco Mor14Rosaria Benedetti15Lucia Altucci16Stefania Butini17Gabriele Carullo18Sandra Gemma19Giuseppe Campiani20Dipartimento di Biotecnologie, Chimica e Farmacia, Università degli Studi di Siena, Via Aldo Moro 2, 53100, Siena, ItalyDipartimento di Scienze della Vita, Università degli Studi di Siena, Via Aldo Moro 2, 53100, Siena, ItalyDipartimento di Biotecnologie, Chimica e Farmacia, Università degli Studi di Siena, Via Aldo Moro 2, 53100, Siena, ItalyDipartimento di Medicina di Precisione, Università della Campania “Luigi Vanvitelli”, Via de Crecchio 7, 80138, Naples, ItalyDipartimento di Scienze degli Alimenti e del Farmaco, Università di Parma, Parco Area delle scienze 27/A, Campus Universitario, 43124, Parma, ItalyDipartimento di Scienze della Vita, Università degli Studi di Siena, Via Aldo Moro 2, 53100, Siena, ItalyDipartimento di Scienze della Vita, Università degli Studi di Siena, Via Aldo Moro 2, 53100, Siena, ItalyDipartimento di Scienze della Vita, Università degli Studi di Siena, Via Aldo Moro 2, 53100, Siena, ItalyDipartimento di Scienze della Vita, Università degli Studi di Siena, Via Aldo Moro 2, 53100, Siena, ItalyDipartimento di Medicina Traslazionale, Università di Ferrara, Via Luigi Borsari 46, 44121, Ferrara, ItalyDipartimento di Medicina Traslazionale, Università di Ferrara, Via Luigi Borsari 46, 44121, Ferrara, ItalyDipartimento di Medicina Traslazionale, Università di Ferrara, Via Luigi Borsari 46, 44121, Ferrara, ItalyDipartimento di Medicina Traslazionale, Università di Ferrara, Via Luigi Borsari 46, 44121, Ferrara, ItalyDipartimento di Scienze degli Alimenti e del Farmaco, Università di Parma, Parco Area delle scienze 27/A, Campus Universitario, 43124, Parma, ItalyDipartimento di Scienze degli Alimenti e del Farmaco, Università di Parma, Parco Area delle scienze 27/A, Campus Universitario, 43124, Parma, Italy; Microbiome Research Hub, University of Parma, Parma, ItalyDipartimento di Medicina di Precisione, Università della Campania “Luigi Vanvitelli”, Via de Crecchio 7, 80138, Naples, Italy; Program of Medical Epigenetics, Vanvitelli Hospital, 80138, Naples, ItalyDipartimento di Medicina di Precisione, Università della Campania “Luigi Vanvitelli”, Via de Crecchio 7, 80138, Naples, Italy; Program of Medical Epigenetics, Vanvitelli Hospital, 80138, Naples, Italy; Biogem Institute of Molecular and Genetic Biology, 83031, Ariano Irpino, ItalyDipartimento di Biotecnologie, Chimica e Farmacia, Università degli Studi di Siena, Via Aldo Moro 2, 53100, Siena, ItalyDipartimento di Biotecnologie, Chimica e Farmacia, Università degli Studi di Siena, Via Aldo Moro 2, 53100, Siena, Italy; Corresponding author.Dipartimento di Biotecnologie, Chimica e Farmacia, Università degli Studi di Siena, Via Aldo Moro 2, 53100, Siena, Italy; Corresponding author.Dipartimento di Biotecnologie, Chimica e Farmacia, Università degli Studi di Siena, Via Aldo Moro 2, 53100, Siena, ItalyIdiopathic pulmonary fibrosis (IPF) is a progressive and debilitating interstitial lung disease characterized by limited therapeutic options, with only two FDA-approved palliative agents currently available. Given its poor prognosis and the high incidence of lung transplantation, there is a pressing need to develop innovative and effective therapeutics. Histone deacetylase 6 (HDAC6) has been identified as a key driver of fibrotic progression in IPF, and selective inhibitors of this isoform were able to revert the fibrotic phenotypes. Proteolysis targeting chimeras (PROTACs) are heterobifunctional molecules that can trigger the degradation of a specific target in cells, including HDAC6 enzyme. Therefore, the application of PROTAC technology may represent a novel therapeutic strategy for IPF. We showcase the design, synthesis, and biological evaluation of a library of first-in-class antifibrotic HDAC6-targeting PROTACs, incorporating our in-house inhibitor 1 as the HDAC6 ligand. Newly developed PROTACs 5a and 5c showed effective degradation of HDAC6 in A549 lung cells and IMR-90 lung fibroblasts. 5a and 5c exhibited significant antifibrotic effects against the TGF-β1 induced fibrotic phenotype on IMR-90 cells, a model that mimics IPF conditions. The generation of putative ternary complexes involving PROTAC molecules, the E3-ligase cereblon (CRBN) and the HDAC6 target protein was supported by molecular modelling techniques, including protein-protein docking and molecular dynamics simulations. Mechanistic investigations, based on pull-down experiments, confirmed that the newly synthetized compounds were able to reduce HDAC6 levels through a proteasome- and CRBN-dependent mechanism as confirmed by experiments with neddylation and proteasome inhibitors. This pioneering exploration of targeted protein degradation in IPF-like conditions provides compelling evidence of its therapeutic promise, paving the way for a broader application of PROTACs in treating rare diseases.http://www.sciencedirect.com/science/article/pii/S2772417425000329HDAC6PROTACsPulmonary fibrosisTargeted protein degradationEpigenetics
spellingShingle Ilaria Cursaro
Valerio Ciccone
Valeria Tudino
Chiara Papulino
Laura Scalvini
Sandra Donnini
Lucia Morbidelli
Luca Bini
Claudia Landi
Chiara Contri
Silvia Pasquini
Fabrizio Vincenzi
Katia Varani
Alessio Lodola
Marco Mor
Rosaria Benedetti
Lucia Altucci
Stefania Butini
Gabriele Carullo
Sandra Gemma
Giuseppe Campiani
Reversing TGF-β1-induced fibrotic phenotype in human lung fibroblasts using a PROTAC tool derived from an indoline-based HDAC6 inhibitor
European Journal of Medicinal Chemistry Reports
HDAC6
PROTACs
Pulmonary fibrosis
Targeted protein degradation
Epigenetics
title Reversing TGF-β1-induced fibrotic phenotype in human lung fibroblasts using a PROTAC tool derived from an indoline-based HDAC6 inhibitor
title_full Reversing TGF-β1-induced fibrotic phenotype in human lung fibroblasts using a PROTAC tool derived from an indoline-based HDAC6 inhibitor
title_fullStr Reversing TGF-β1-induced fibrotic phenotype in human lung fibroblasts using a PROTAC tool derived from an indoline-based HDAC6 inhibitor
title_full_unstemmed Reversing TGF-β1-induced fibrotic phenotype in human lung fibroblasts using a PROTAC tool derived from an indoline-based HDAC6 inhibitor
title_short Reversing TGF-β1-induced fibrotic phenotype in human lung fibroblasts using a PROTAC tool derived from an indoline-based HDAC6 inhibitor
title_sort reversing tgf β1 induced fibrotic phenotype in human lung fibroblasts using a protac tool derived from an indoline based hdac6 inhibitor
topic HDAC6
PROTACs
Pulmonary fibrosis
Targeted protein degradation
Epigenetics
url http://www.sciencedirect.com/science/article/pii/S2772417425000329
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