Trio-Drug Combination of Sodium Valproate, Baclofen and Thymoquinone Exhibits Synergistic Anticonvulsant Effects in Rats and Neuro-Protective Effects in HEK-293 Cells

Abstract

Epilepsy is a chronic brain disorder, with anti-epileptic drugs (AEDs) providing relief from hyper-excitability of neurons, but largely failing to restrain neurodegeneration. We investigated a progressive preclinical trial in rats, whereby the test drugs; sodium valproate (SVP; 150 and 300 mg/kg), baclofen (BFN; 5 and 10 mg/kg), and thymoquinone (THQ; 40 and 80 mg/kg) were administered (i.p, once/day for 15 days) alone, and as low dose combinations, and subsequently tested for antiseizure and neuroprotective potential using electrical stimulation of neurons by Maximal electroshock (MES). The seizure stages were monitored, and hippocampal levels of m-TOR, IL-1β, IL-6 were measured. Hippocampal histopathology was also performed. Invitro and Insilco studies were run to counter-confirm the results from rodent studies. We report the synergistic effect of trio-drug combination; SVP (150 mg/kg), BFN (5 mg/kg) and THQ (40 mg/kg) against generalized seizures. The Insilco results revealed that trio-drug combination binds the Akt active site as a supramolecular complex, which could have served as a delivery system that affects the penetration and the binding to the new target. The potential energy of the ternary complex in the Akt active site after dynamics simulation was found to be −370.426 Kcal/mol, while the supramolecular ternary complex alone was −38.732 Kcal/mol, with a potential energy difference of −331.694 Kcal/mol, which favors the supramolecular ternary complex at Akt active site binding. In addition, the said combination increased cell viability by 267% and reduced morphological changes induced by Pentylenetetrazol (PTZ) in HEK-293 cells, which indicates the neuroprotective property of said combination. To conclude, we are the first to report the anti-convulsant and neuroprotective potential of the trio-drug combination.

Repetitive transcranial magnetic stimulation promotes motor function recovery in mice after spinal cord injury via regulation of the Cx43-autophagy loop

“…After SCI, an inhibitory microenvironment is formed in the lesion area, and adult neurons exhibit poor intrinsic growth potential [ 23 , 24 ]. After SCI, autophagic flux is impaired, and the accumulation of a large number of oxidative stress products accelerates apoptosis [ 10 , 11 ]. Furthermore, Cx43 has been shown to protect embryos from reactive oxygen species (ROS)-induced autophagic stress and apoptosis to ensure the maintenance of mitochondrial membrane potential and ATP production, while baclofen can promote protective autophagy by downregulating Cx43 expression on cell and mitochondrial membranes [ 25 ].…”

“…Autophagic flux is impaired after SCI, and the dynamic balance of autophagic flow is disrupted, leading to autophagy stress and the accumulation of numerous oxidative stress products to accelerate cell apoptosis [ 9 ]. Recent studies have demonstrated that gap junction protein 43 (connexin43, Cx43) protects embryos from reactive oxygen species (ROS)-induced autophagic stress and apoptosis, maintaining the mitochondrial membrane potential and production of ATP [ 10 ], while baclofen can enhance protective autophagy [ 11 ] by downregulating the expression of Cx43 on cells and mitochondrial surfaces. These findings indicate that Cx43 regulates autophagy during oxidative stress.…”

Nigella sativa and its nano‐mediated approach toward management of neurodegenerative disorders: A review

“…The Insilco results revealed that trio-drug combination binds the Akt active site as a supramolecular complex, which could have served as a delivery system that affects the penetration and the binding to the new target [61] Abbreviations: AKT, protein kinase B; BFN, baclofen; Htf, human transferrin; mTOR, mammalian target of rapamycin; PHT, phenytoin; SERT, serotonin reuptake transporter; SVP, sodium valproate.…”

Modulation of GABAB receptors in the insula bidirectionally affects associative memory of epilectic rats in both spatial and non-spatial operant tasks

“…Furthermore, BLF showed an anti-convulsant effect in pentylenetetrazol-induced model of epilepsy in developing rats of 7, 12, 18, and 25 days ( Kubová et al, 1996 ). In adult rats, BLF reduced pentylenetetrazol-induced seizures ( Chen et al, 2004 ) and electroshock-induced seizures ( Hyder Pottoo et al, 2022 ). Similarly, BLF reduced seizures in a mouse pentylenetetrazole kindling model of epilepsy ( De Sarro et al, 2000 ).…”