title: Thiorphan, a neutral endopeptidase inhibitor used for diarrhoea, is neuroprotective in newborn mice.
abstract: Excitotoxic damage appears to be a critical factor in the formation of perinatal brain lesions associated with cerebral palsy (CP). When injected into newborn mice, the glutamatergic analogue, ibotenate, produces cortical lesions and white matter cysts that mimic human perinatal brain lesions. Neuropeptides are neuronal activity modulators and could therefore modulate glutamate-induced lesions. However, neuropeptides are rapidly degraded by peptidases. Racecadotril, which is rapidly metabolized to its active metabolite thiorphan, is a neutral endopeptidase (NEP) inhibitor used in clinical practice for diarrhoea with a remarkable safety profile. This study aimed to test the original hypothesis that thiorphan could be neuroprotective against ibotenate-induced lesions in newborn mice. Intraperitoneal administration of thiorphan reduced ibotenate-induced cortical lesions by up to 57% and cortical caspase-3 cleavage by up to 59%. This neuroprotective effect was long-lasting and was still observed when thiorphan was administered 12 h after the insult, showing a remarkable window for therapeutic intervention. Further supporting the neuroprotective effect of pharmacological blockade of NEP, mouse pups with a genetic deletion of NEP displayed a significantly reduced size of the ibotenate-induced cortical grey matter lesion when compared with wild-type animals. Thiorphan effects were mimicked by substance P (SP) and, in a less potent manner, by neurokinin A. Thiorphan effects were inhibited by blockers of NK1 and NK2 receptors. Real-time reverse transcription-polymerase chain reaction, autoradiography and immunohistochemistry confirmed the expression of NK1 and NK2 receptors in the neonatal murine neocortex. These data demonstrate that thiorphan prevents neonatal excitotoxic cortical damage, an effect largely mediated by SP. Thiorphan could represent a promising drug for the prevention of CP, which remains a challenging disease. In a broader context, these results also raise potential implications for the prevention of neurodegenerative diseases involving glutamate-mediated excitotoxic neuronal death.
authors.affiliation: Inserm U676, Service de Neurologie Pédiatrique, Paris, France.
authors.forename: Fadia
authors.initials: F
authors.lastname: Medja
authors.forename: Vincent
authors.initials: V
authors.lastname: Lelièvre
authors.forename: Romain H
authors.initials: RH
authors.lastname: Fontaine
authors.forename: Fanny
authors.initials: F
authors.lastname: Lebas
authors.forename: Philippe
authors.initials: P
authors.lastname: Leroux
authors.forename: Tanja
authors.initials: T
authors.lastname: Ouimet
authors.forename: Alois
authors.initials: A
authors.lastname: Saria
authors.forename: Catherine
authors.initials: C
authors.lastname: Rougeot
authors.forename: Pascal
authors.initials: P
authors.lastname: Dournaud
authors.forename: Pierre
authors.initials: P
authors.lastname: Gressens
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D053148:Caspase 3 / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000187:drug effects* / Q000473:pathology; D002547:Cerebral Palsy / Q000473:pathology* / Q000517:prevention & control; D018690:Excitatory Amino Acid Agonists; D005260:Female; D007051:Ibotenic Acid; D007150:Immunohistochemistry / Q000379:methods; D007274:Injections, Intraperitoneal; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D015260:Neprilysin / Q000037:antagonists & inhibitors*; D015288:Neurokinin A / Q000494:pharmacology; D064729:Neurokinin-1 Receptor Antagonists; D009474:Neurons / Q000187:drug effects; D018696:Neuroprotective Agents / Q000008:administration & dosage*; D018377:Neurotransmitter Agents / Q000494:pharmacology; D011480:Protease Inhibitors / Q000008:administration & dosage*; D018041:Receptors, Neurokinin-2 / Q000037:antagonists & inhibitors; D012640:Seizures / Q000517:prevention & control; D013373:Substance P / Q000494:pharmacology; D015244:Thiorphan / Q000008:administration & dosage*
chemical_list: D018690:Excitatory Amino Acid Agonists; D064729:Neurokinin-1 Receptor Antagonists; D018696:Neuroprotective Agents; D018377:Neurotransmitter Agents; D011480:Protease Inhibitors; D018041:Receptors, Neurokinin-2; D007051:Ibotenic Acid; D013373:Substance P; D015288:Neurokinin A; D015244:Thiorphan; D053148:Caspase 3; D015260:Neprilysin