Calpain inhibitors and antioxidants act synergistically to prevent cell necrosis: effects of the novel dual inhibitors (cysteine protease inhibitor and antioxidant) BN 82204 and its pro‐drug BN 82270

Abstract

Cell death is a common feature observed in neurodegenerative disorders, and is often associated with calpain activation and overproduction of reactive oxygen species (ROS). This study investigated the use of calpain inhibitors and antioxidants in combination to protect cells against necrosis. Maitotoxin (MTX), which induces a massive influx of calcium, was used to provoke neuronal cell death. This toxin increased, in a concentration-dependent manner, both calpain activity and ROS formation. Calpain inhibitors or antioxidants inhibited MTX-induced necrosis only marginally (below 20%), whereas their association protected against cell death by 40-66% in a synergistic manner. BN 82204, which possesses both calpain-cathepsin L inhibitory and antioxidant properties, and its acetylated pro-drug BN 82270, totally protected cells at 100 lM. The pro-drug BN 82270, which had better cell penetration, was twice as effective as the active principle BN 82204 in protecting glioma C6 or neuroblastoma SHSY5Y cells against death. These results suggest the potential therapeutic relevance of using a single molecule with multiple activities (cysteine protease inhibitor/antioxidant), and warrant further in vivo investigations in models of neuronal disorders.

From Cochlear Cell Death Pathways To New Pharmacological Therapies

“…Consistent with this, when the calpain inhibitor leupeptin was infused into the basal turn of the chinchilla cochlea, it reduced sensory hair cell loss following a 105 dB SPL exposure by as much as 60% [99]. In addition, infusion of BN82270, a novel dual inhibitor of calpains and of lipid peroxidation [151] into the scala tympani of the guinea pig cochlea prevented the formation of calpain-cleaved fodrin, DNA fragmentation and hair cell degeneration caused by sound trauma. This was confirmed by functional tests in vivo, showing a clear dose-dependent reduction of permanent hearing loss.…”

Calpains and neuronal damage in the ischemic brain: The swiss knife in synaptic injury

“…Compounds 11 to 14 (Abell et al, 2005;Abell et al, 2009;Tyndall et al, 2005) (Bihovsky et al, 2004;Cuerrier et al, 2005;Donkor et al, 2008;Donkor et al, 2001) Peptidomimetics with enhanced pharmacokinetic profiles Compounds 49 to 51, SNJ-1715, 52 SNJ-1757, 53, Amphiphile and 54 SNJ-1945 (Nakamura and Inoue, 2002;Nakamura et al, 2003;Shirasaki et al, 2005;Shirasaki et al, 2006) Site specific targeting of calpain inhibitors Compounds 55 to 60 (Lescop et al, 2005) Dual function calpain inhibitors BN-82204 61, BN-82270 62, 63 (Auvin et al, 2004;Fareed et al, 2006;Pignol et al, 2006) …”

The Mitochondrial Complex I Inhibitor Rotenone Induces Endoplasmic Reticulum Stress and Activation of GSK-3β in Cultured Rat Retinal Cells

“…calpain-l contributed to nonapoptotic neuronal death but not to apoptotic glial cell death. Indeed, the prevention of cell death by calpain inhibition has been demonstrated in human neuroblastoma 49 and hypoxic-ischemic brain injury, 50 where nonapoptotic modes of cell death are involved. Accordingly, one limitation of the present investigations is that we did not determine the proportion of cell death due to necrosis.…”