Neuroprotective Effect of 3-(Naphthalen-2-Yl(Propoxy)Methyl)Azetidine Hydrochloride on Brain Ischaemia/Reperfusion Injury

Because ischaemic stroke is one of the most common brain disorders, diverse effective therapies are urgently required. Recent studies reported a variety of azetidine-based scaffolds for the development of central nervous system-focused lead-like libraries. However, their mechanisms of action and in vivo functions remain unclear. Here, we investigated the potential mechanism and beneficial effects of 3-(naphthalen-2-yl(propoxy)methyl)azetidine hydrochloride (KHG26792), a novel azetidine derivative, on ischaemia/reperfusion (I/R) brain injury. We adapted a mouse brain ischaemia model induced by 2 h of middle cerebral artery occlusion followed by 24 h of reperfusion. We measured apoptotic cell death, inflammatory mediators, free radical generation, and anti-oxidative enzymes activities. We also measured the mitochondrial ATP level and Na+, K+-ATPase and cytochrome c oxidase activities. Using western blotting, we analysed the protein levels of inducible NOS, hypoxia-upregulated protein 1, PTEN-induced putative kinase, uncoupling protein 2, p-Akt, MMP-3, and full-length receptor for advanced glycation end-products (RAGE). KHG26792 significantly improved neurological deficits and brain oedema and suppressed I/R-induced apoptosis. KHG26792 significantly attenuated I/R-induced inflammation and oxidative stress by upregulating SOD and catalase activity, GSH, p-Akt, mitochondrial ATP, Na+, K+-ATPase, cytochrome c oxidase, and soluble RAGE and downregulating iNOS, HYOUP1, and MMP-3, suggesting a potential anti-inflammatory and antioxidant role of KHG26792. This is the first study to show that KHG26792 can protect mouse brains against I/R injury by inhibiting apoptotic damage, modulating inflammation, scavenging free radicals, ameliorating oxidative stress, and improving the energy metabolism of the brain, although the clinical relevance of our findings remains unknown.