Genuine selective caspase-2 inhibition with new irreversible small peptidomimetics
Bosc et al. Cell Death and Disease (2022) 13:959 (Impact Factor 8.1)
Caspase-2 (Casp2) is a promising therapeutic target in several human diseases, including nonalcoholic steatohepatitis (NASH) and Alzheimer’s disease (AD). However, the design of an active-site-directed inhibitor selective to individual caspase family members is challenging because caspases have extremely similar active sites. Here we present new peptidomimetics derived from the VDVAD pentapeptide structure, harboring non-natural modifications at the P2 position and an irreversible warhead. Enzyme kinetics show that these new compounds, such as LJ2 or its specific isomers LJ2a, and LJ3a, strongly and irreversibly inhibit Casp2 with genuine selectivity. In agreement with the established role of Casp2 in cellular stress responses, LJ2 inhibits cell death induced by microtubule destabilization or hydroxamic acid-based deacetylase inhibition. The most potent peptidomimetic, LJ2a, inhibits human Casp2 with a remarkably high inactivation rate (k3/Ki ~5,500,000 M−1 s−1), and the most selective inhibitor, LJ3a, has close to a 1000 times higher inactivation rate on Casp2 as compared to Casp3. Structural analysis of LJ3a shows that the spatial configuration of Cα at the P2 position determines inhibitor efficacy. In transfected human cell lines overexpressing site-1 protease (S1P), sterol regulatory element-binding protein 2 (SREBP2) and Casp2, LJ2a and LJ3a fully inhibit Casp2-mediated S1P cleavage and thus SREBP2 activation, suggesting a potential to prevent NASH development. Furthermore, in primary hippocampal neurons treated with β-amyloid oligomers, submicromolar concentrations of LJ2a and of LJ3a prevent synapse loss, indicating a potential for further investigations in AD treatment.
Elodie Bosc1, Julie Anastasie1, Feryel Soualmia1, Pascale Coric 2, Ju Youn Kim3, Lily Q. Wang3, Gullen Lacin1,4, Kaitao Zhao 5,6, Ronak Patel5,6, Eric Duplus1, Philippe Tixador1, Andrew A. Sproul5,6, Bernard Brugg1, Michelle Reboud-Ravaux1, Carol M. Troy 5,6,7, Michael L. Shelanski5,6, Serge Bouaziz2, Michael Karin3, Chahrazade El Amri1 and Etienne D. Jacotot 1,5,6
1 INSERM U1164, CNRS UMR 8256, Sorbonne Université, Campus Pierre et Marie Curie, Paris F-75005, France. 2 Université de Paris, CNRS, CiTCoM, F-75006 Paris, France. 3 Laboratory of Gene Regulation and Signal Transduction, Departments of Pharmacology and Pathology, University of California San Diego, School of Medicine, La Jolla, CA 92093, USA. 4 MicroBrain Biotech S.A.S. 52 Avenue de l’Europe, Marly-Le-Roi F-78160, France. 5 Taub Institute for Research on Alzheimer’s Disease and the Aging Brain, Columbia University, New York, NY, USA. 6 Department of Pathology and Cell Biology, Columbia University, New York, NY, USA. 7 Department of Neurology, Columbia University, New York, NY, USA.
Received: 3 December 2021 Revised: 26 October 2022 Accepted: 1 November 2022
Cell Death and Disease (2022) 13:959