Repurposing FDA Approved Drugs as JNK3 Inhibitor for Prevention of Neuroinflammation Induced by MCAO in Rats

Abstract

Background Stress-associated kinases are considered major pathological mediators in several incurable neurological disorders. Importantly, among these stress kinases, the c-Jun NH2-terminal kinase (JNK) has been linked to numerous neuropathological conditions, including oxidative stress, neuroinflammation, and brain degeneration associated with brain injuries such as ischemia/reperfusion injury. In this study, we adopted a drug repurposing/reprofiling approach to explore novel JNK3 inhibitors from FDA-approved medications to supplement existing therapeutic strategies. Materials and Methods We performed in silico docking analysis and molecular dynamics simulation to screen potential candidates from the FDA approved drug library using the standard JNK inhibitor SP600125 as a reference. After the virtual screening, dabigatran, estazolam, leucovorin, and pitavastatin were further examined in ischemic stroke using an animal rodent model of focal cerebral ischemia using transient middle cerebral artery occlusion (t-MCAO). The selected drugs were probed for neuroprotective effectiveness by measuring the infarct area (%) and neurological deficits using a 28-point composite score. Biochemical assays including ELISA and immunohistochemical experiments were performed. Results We obtained structural insights for dabigatran, estazolam, and pitavastatin binding to JNK3, revealing a significant contribution of the hydrophobic regions and significant residues of active site regions. To validate the docking results, the pharmacological effects of dabigatran, estazolam, leucovorin, and pitavastatin on MCAO were tested in parallel with the JNK inhibitor SP600125. After MCAO surgery, severe neurological deficits were detected in the MCAO group compared with the sham controls, which were significantly reversed by dabigatran, estazolam, and pitavastatin treatment. Aberrant morphological features and brain damage were observed in the ipsilateral cortex and striatum of the MCAO groups. The drugs restored the anti-oxidant enzyme activity and reduced the levels of oxidative stress-induced p-JNK and neuroinflammatory mediators such as NF-kB and TNF-ɑ in rats subjected to MCAO. Conclusion Our results demonstrated that the novel FDA-approved medications attenuate ischemic stroke-induced neuronal degeneration, possibly by inhibiting JNK3. Being FDA-approved safe medications, the use of these drugs can be clinically translated for ischemic stroke-associated brain degeneration and other neurodegenerative diseases associated with oxidative stress and neuroinflammation.

Natural Dietary Supplement, Carvacrol, Alleviates LPS-Induced Oxidative Stress, Neurodegeneration, and Depressive-Like Behaviors via the Nrf2/HO-1 Pathway

“…Moreover, the JNK pathway can be activated by multiple factors, including ROS and proinflammatory mediators. 68 Likewise, a close interplay of caspase-3 and JNK is implicated in the execution of the mitochondrial apoptotic pathway in several neurodegenerative disorders. 69,70 Our data demonstrated a considerably decreased expression of anti-apoptotic Bcl2 as well as a significant increase in the expression of JNK and caspase-3 in LPS-injected rats.…”

Carveol Attenuates Seizure Severity and Neuroinflammation in Pentylenetetrazole-Kindled Epileptic Rats by Regulating the Nrf2 Signaling Pathway

“…We mixed 0.1 mL of tissue homogenate with 0.1 mL of pyrogallol solution (1 M) and 2.8 mL of 0.1 M potassium phosphate buffer (pH 7.4), which yielded a reaction mixture of 3 mL. The absorbance was measured at 312 nm [ 51 ]. SOD activity was expressed in U/mg of protein.…”

“…The absorbance of the final mixture was measured at a wavelength of 240 nm. The catalase activity was calculated using the following formula: where δ O.D represents the change in absorbance per minute and E represents the extinction coefficient of H 2 O 2 with a value of 0.071 mmol cm −1 [ 51 ]. The Lowery method was used to measure protein levels.…”

Suppression of LPS-Induced Inflammation and Cell Migration by Azelastine through Inhibition of JNK/NF-κB Pathway in BV2 Microglial Cells

“…For example, elevated expressions of JNK3, c-Jun, as well as levels of pro-inflammatory cytokines, including TNF-α, IL-6, and IL-1β, were observed in the hippocampus of the chronic social defeat stress mice model [42]. Moreover, in rats subjected to transient middle cerebral artery occlusion, several JNK3 inhibitors are shown to restore anti-oxidant enzyme activity and reduce the levels of oxidative stressinduced phosphorylation of JNK and neuroinflammatory mediators [43]. These reports strongly suggest that inhibition of JNK3 can be a favorable strategy to alleviate the progression of neuroinflammation and neurodegeneration.…”