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.
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 microm. 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.
Pignol
Bernadette
B
Department of Neurobiology, Ipsen Research Laboratories, Les Ulis, France. Bernadette.pignol@ipsen.com
Auvin
Serge
S
Carré
Denis
D
Marin
Jean-Grégoire
JG
Chabrier
Pierre-Etienne
PE
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D002154:Calpain / Q000037:antagonists & inhibitors* / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology*; D015237:Dinoprost / Q000031:analogs & derivatives / Q000378:metabolism; D004357:Drug Synergism; D006801:Humans; D015227:Lipid Peroxidation / Q000187:drug effects; D008727:Methotrexate / Q000494:pharmacology; D009336:Necrosis; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D010640:Phenothiazines / Q000494:pharmacology*; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012697:Serine Endopeptidases / Q000494:pharmacology
D000975:Antioxidants; C513568:BN 82204; D015853:Cysteine Proteinase Inhibitors; D019384:Nucleic Acid Synthesis Inhibitors; D010640:Phenothiazines; C075750:8-epi-prostaglandin F2alpha; D015237:Dinoprost; D012697:Serine Endopeptidases; D002154:Calpain; C484806:CAPN1 protein, human; D046988:Proteasome Endopeptidase Complex; D008727:Methotrexate