Identification of ER-000444793, a Cyclophilin D-independent inhibitor of mitochondrial permeability transition, using a high-throughput screen in cryopreserved mitochondria

• DOI: 10.1038/srep37798
• Publication Date: 2016-11-25T10:34:56Z
• Authors: Thomas Briston; Sian Lewis; Mumta Koglin; Kavita Mistry; Yongchun Shen; Naomi Hartopp; Ryosuke Katsumata; Hironori Fukumoto; Michael R. Duchen; Gyorgy Szabadkai; James M. Staddon; Malcolm Roberts; Ben Powney
• Keywords: MPTP; Voltage-dependent anion channel

Abstract Growing evidence suggests persistent mitochondrial permeability transition pore (mPTP) opening is a key pathophysiological event in cell death underlying variety of diseases. While it has long been clear the mPTP druggable target, current agents are limited by off-target effects and low therapeutic efficacy. Therefore identification development novel inhibitors necessary. To rapidly screen large compound libraries for modulators, method was exploited to cryopreserve batches functionally active mitochondria from cells tissues. The cryopreserved maintained respiratory coupling ATP synthesis, Ca 2+ uptake transmembrane potential. A high-throughput (HTS), using an assay -induced swelling identified ER-000444793, potent inhibitor opening. Further evaluation assays membrane depolarisation retention capacity also indicated that ER-000444793 acted as mPTP. neither affected cyclophilin D (CypD) enzymatic activity, nor displaced CsA CypD protein, suggesting mechanism independent inhibition. Here we novel, CypD-independent screening approach described provides workflow additional tool aid search modulators help understand its molecular nature.