Department of Pharmacology, College of Medicine, The University of Arizona, Tucson, AZ, United States.
Department of Pharmacology and Toxicology, College of Pharmacy, The University of Arizona, Tucson, AZ, United States.
Pain. 2020 Nov;161(11):2551-2570. doi: 10.1097/j.pain.0000000000001955.
The voltage-gated calcium channels CaV3.1-3.3 constitute the T-type subfamily, whose dysfunctions are associated with epilepsy, psychiatric disorders, and chronic pain. The unique properties of low-voltage-activation, faster inactivation, and slower deactivation of these channels support their role in modulation of cellular excitability and low-threshold firing. Thus, selective T-type calcium channel antagonists are highly sought after. Here, we explored Ugi-azide multicomponent reaction products to identify compounds targeting T-type calcium channel. Of the 46 compounds tested, an analog of benzimidazolonepiperidine-5bk (1-{1-[(R)-{1-[(1S)-1-phenylethyl]-1H-1,2,3,4-tetrazol-5-yl}(thiophen-3-yl)methyl]piperidin-4-yl}-2,3-dihydro-1H-1,3-benzodiazol-2-one) modulated depolarization-induced calcium influx in rat sensory neurons. Modulation of T-type calcium channels by 5bk was further confirmed in whole-cell patch clamp assays in dorsal root ganglion (DRG) neurons, where pharmacological isolation of T-type currents led to a time- and concentration-dependent regulation with a low micromolar IC50. Lack of an acute effect of 5bk argues against a direct action on T-type channels. Genetic knockdown revealed CaV3.2 to be the isoform preferentially modulated by 5bk. High voltage-gated calcium, as well as tetrodotoxin-sensitive and -resistant sodium, channels were unaffected by 5bk. 5bk inhibited spontaneous excitatory postsynaptic currents and depolarization-evoked release of calcitonin gene-related peptide from lumbar spinal cord slices. Notably, 5bk did not bind human mu, delta, or kappa opioid receptors. 5bk reversed mechanical allodynia in rat models of HIV-associated neuropathy, chemotherapy-induced peripheral neuropathy, and spinal nerve ligation-induced neuropathy, without effects on locomotion or anxiety. Thus, 5bk represents a novel T-type modulator that could be used to develop nonaddictive pain therapeutics.
电压门控钙通道 CaV3.1-3.3 构成 T 型亚家族,其功能障碍与癫痫、精神障碍和慢性疼痛有关。这些通道的低电压激活、更快失活和更慢失活的独特特性支持它们在调节细胞兴奋性和低阈值放电中的作用。因此,选择性 T 型钙通道拮抗剂备受关注。在这里,我们探索了 Ugi-叠氮化物多组分反应产物,以鉴定针对 T 型钙通道的化合物。在所测试的 46 种化合物中,苯并咪唑啉酮哌啶-5bk(1-{1-[(R)-{1-[(1S)-1-苯乙基]-1H-1,2,3,4-四唑-5-基}(噻吩-3-基)甲基]哌啶-4-基}-2,3-二氢-1H-1,3-苯并二氮杂-2-酮)模拟了大鼠感觉神经元中去极化诱导的钙内流。5bk 对 T 型钙通道的调节在背根神经节 (DRG) 神经元的全细胞膜片钳实验中得到进一步证实,其中 T 型电流的药理学分离导致时间和浓度依赖性调节,具有低微摩尔 IC50。5bk 缺乏急性作用表明其对 T 型通道没有直接作用。基因敲低显示 CaV3.2 是被 5bk 优先调节的亚型。高电压门控钙通道以及河豚毒素敏感和耐受的钠通道不受 5bk 影响。5bk 抑制腰椎脊髓切片中的自发性兴奋性突触后电流和降钙素基因相关肽的去极化诱发释放。值得注意的是,5bk 不与人类 mu、delta 或 kappa 阿片受体结合。5bk 逆转了 HIV 相关神经病、化疗引起的周围神经病和脊神经结扎引起的神经病模型中的机械性痛觉过敏,而对运动或焦虑没有影响。因此,5bk 代表一种新型 T 型调节剂,可用于开发非成瘾性疼痛治疗药物。