LAUSR

Mitochondrial dysfunction is associated with cerebral ischemic stroke. Sovateltide (IRL-1620; PMZ-1620) undergoing clinical trials (NCT04047563; NCT04046484) has been demonstrated, in preclinical studies, to provide recovery of brain functions when administered following acute cerebral ischemic stroke. We hypothesize that sovateltide preserves mitochondrial activity and provides neural protection and regeneration. We investigated the effect of sovateltide on mitochondrial morphology and function following acute cerebral ischemic stroke. Rats underwent permanent middle cerebral artery occlusion (MCAO) and then received three intravenous injections of vehicle or sovateltide (5 μg/kg) at 4, 6, and 8 h and sacrificed at 24 h post MCAO. Another set of rats were treated similarly on day 0, 3 and 6 and sacrificed on day 7. Western blot and/or immunofluorescence were used to evaluate protein expression of NeuroD1, DoubleCortin, HUC+HUD, NeuN, Sox2, Oct4, DRP1 and MFN2. Mitochondrial biogenesis was evaluated using in situ PCR and transmission electron microscopy (TEM). Sovateltide produced (p<0.001) improvement in neurological deficit and motor function compared to vehicle at 24 h and day 7 post MCAO. Expression of NeuroD1 (p=0.0003), HuC-HuD (p=0.0373) and DoubleCortin (p=0.013) was higher in sovateltide compared to vehicle at 24 h but not at day 7. No change in Oct4 and Sox2 was observed. Immunofluorescence of cultured NPCs showed elevated expression of NeuroD1 and NeuN in sovateltide compared to vehicle. Sovateltide significantly decreased fission marker, DRP1 (p<0.001) and increased fusion marker MFN2 (p<0.0001) at 24 h and day 7 compared to vehicle. A significantly increased mitochondrial DNA (MT-ATP8, mitochondria specific gene) fluorescence in sovateltide group compared to vehicle (p=0.0418) and sham (p=0.0085) was observed. TEM analysis showed significant increase in mitochondrial cross-sectional area x number and % mitochondrial/tissue area in sovateltide group at 24 hours compared to vehicle. Mitochondrial number increased (p=0.032) at day 7 compared to 24 h in vehicle group. Sovateltide preserves mitochondrial activity and provides neural protection and regeneration in rat model of acute cerebral ischemic stroke.