Involvement of selective GABA-A receptor subtypes in amelioration of cisplatin-induced neuropathic pain by 2′-chloro-6-methyl flavone (2′-Cl-6MF)

Cisplatin-induced peripheral neuropathic pain is a common adverse effect of chemotherapy. The present study evaluated the effects of 2'-chloro-6-methylflavone (2'-Cl-6MF) at recombinant alpha 1 beta 2 gamma 2L, alpha 2 beta 1-3 gamma 2L, and alpha 3 beta 1-3 gamma 2L GABA-A receptor subtypes expressed in Xenopus oocytes and subsequently evaluated its effectiveness in cisplatin-induced neuropathic pain. The results showed that 2'-Cl-6MF potentiated GABA-elicited currents at alpha 2 beta 2/3 gamma 2L and alpha 3 beta 2/3 gamma 2L GABA-A receptor subtypes. The potentiation was blocked by the co-application of flumazenil (a benzodiazepine (BDZs) site antagonist). In behavioral studies, mechanical allodynia was induced by intraplantar injection of cisplatin (40 mu g/paw) in Sprague Dawley rats, and behavioral assessments were made 24 h after injection. 2'-Cl-6MF (1, 10, 30, and 100 mg/kg, i.p.), was administered 1 h before behavioral evaluation. Administration of 2'-Cl-6MF (30 and 100 mg/kg, i.p) significantly enhanced the paw withdrawal threshold and decreased mechanical allodynia. The standard drugs, gabapentin (GBP) at the dose of 70 mg/kg, and HZ 166 (16 mg/kg), i.p. also significantly enhanced the paw withdrawal threshold in mechanical allodynia. Pretreatment with pentylenetetrazole (PTZ) (15 mg/kg, i.p.) and flumazenil reversed the antinociceptive effect of 2'-Cl-6MF in mechanical allodynia indicating GABAergic mechanisms. Moreover, the binding mechanism of 2'-Cl-6MF was rationalized by in silico modeling tools. The 3D-coordinates of alpha 2 beta 2 gamma 2L and alpha 2 beta 3 gamma 2L were generated after homology modeling of the alpha 2 subtype and 2'-Cl-6MF was at predicted binding sites of the developed models. The alpha 2 model was compared with the alpha 1 and alpha 3 subunits via structural and sequence alignment. Molecular docking depicted that the compound binds efficiently at the neuromodulator binding site of the receptors. The findings of this study revealed that 2'-Cl-6MF ameliorated the manifestations of cisplatin-induced neuropathic pain in rats. Furthermore, we also conclude that GABAergic mechanisms may contribute to the antinociceptive effect of 2'-Cl-6MF. The molecular docking studies also confirm the involvement of the BDZs site of GABA-A receptors. It was observed that Ile230 of alpha 2 stabilize the chlorophenyl ring of 2'-Cl-6MF through hydrophobic interactions, which is replaced by Val203 in alpha 1 subunit. However, the smaller side chain of Val203 does not provide hydrophobic interaction to the compound due to high conformational flexibility of alpha 1 subunit.

Automated summary

The study demonstrated that 2'-chloro-6-methylflavone can alleviate cisplatin-induced neuropathic pain and this effect may be associated with GABAergic mechanisms. The molecular docking studies support the involvement of the BDZs site of GABA-A receptors in the mechanism of action.