众所周知,GABAA受体正调节剂可诱导镇静作用。睡眠,和全身麻醉。相反,GABAA受体负变构调节剂(GABAARNAMs)可以增加唤醒并诱发癫痫发作。受我们对失眠症患者的研究的启发,我们发现两个GABAARNAM可以恢复体外兴奋/抑制(E/I)平衡和体内唤醒,我们选择筛选11种已报道可调节唤醒的化合物,看看他们是否共享GABA相关机制。我们用常规和微流体膜片钳方法确定了调制。我们发现受体激活受到所有11种化合物的可变调节:利福平(RIF),甲硝唑(MET),米诺环素(MIN),红霉素(ERY),氧氟沙星(OFX),氯喹(CQ),硫酸羟氯喹(HCQ),氟马西尼(FLZ),戊四氮(PTZ),(-)-表没食子儿茶素没食子酸酯(EGCG),和克拉霉素(CLR)。调节剂-受体相互作用的计算模型预测了药物在典型结合位点和受体上新的孤儿位点的作用。我们的发现表明,现在可以通过多种研究途径来研究大型和脑渗透分子,以治疗CNSE/I平衡减弱的患者。
GABAA receptor-positive modulators are well-known to induce sedation, sleep, and general anesthesia. Conversely, GABAA receptor negative allosteric modulators (GABAARNAMs) can increase arousal and induce seizures. Motivated by our studies with patients with hypersomnia, and our discovery that two GABAARNAMs can restore the Excitation/Inhibition (E/I) balance in vitro and arousal in vivo, we chose to screen 11 compounds that have been reported to modulate arousal, to see if they shared a GABA-related mechanism. We determined modulation with both conventional and microfluidic patch clamp methods. We found that receptor activation was variably modulated by all 11 compounds: Rifampicin (RIF), Metronidazole (MET), Minocycline (MIN), Erythromycin (ERY), Ofloxacin (OFX), Chloroquine (CQ), Hydroxychloroquine sulfate (HCQ), Flumazenil (FLZ), Pentylenetetrazol (PTZ), (-)-Epigallocatechin Gallate (EGCG), and clarithromycin (CLR). The computational modeling of modulator-receptor interactions predicted drug action at canonical binding sites and novel orphan sites on the receptor. Our findings suggest that multiple avenues of investigation are now open to investigate large and brain-penetrant molecules for the treatment of patients with diminished CNS E/I balance.