Xiang Zixiu, Thompson Analisa D, Brogan John T, Schulte Michael L, Melancon Bruce J, Mi Debbie, Lewis L Michelle, Zou Bende, Yang Liya, Morrison Ryan, Santomango Tammy, Byers Frank, Brewer Katrina, Aldrich Jonathan S, Yu Haibo, Dawson Eric S, Li Min, McManus Owen, Jones Carrie K, Daniels J Scott, Hopkins Corey R, Xie Ximin Simon, Conn P Jeffrey, Weaver C David, Lindsley Craig W
Department of Pharmacology, Vanderbilt University Medical Center, Nashville, TN 37232-6600, USA.
ACS Chem Neurosci. 2011 Dec 21;2(12):730-742. doi: 10.1021/cn200090z.
T-type Ca(2+) channel inhibitors hold tremendous therapeutic potential for the treatment of pain, epilepsy, sleep disorders, essential tremor and other neurological disorders; however, a lack of truly selective tools has hindered basic research, and selective tools from the pharmaceutical industry are potentially burdened with intellectual property (IP) constraints. Thus, an MLPCN high-throughput screen (HTS) was conducted to identify novel T-type Ca(2+) channel inhibitors free from IP constraints, and freely available through the MLPCN, for use by the biomedical community to study T-type Ca(2+) channels. While the HTS provided numerous hits, these compounds could not be optimized to the required level of potency to be appropriate tool compounds. Therefore, a scaffold hopping approach, guided by SurflexSim, ultimately afforded ML218 (CID 45115620) a selective T-Type Ca(2+) (Ca(v)3.1, Ca(v)3.2, Ca(v)3.3) inhibitor (Ca(v)3.2, IC(50) = 150 nM in Ca(2+) flux; Ca(v)3.2 IC(50) = 310 nM and Ca(v)3.3 IC(50) = 270 nM, respectively in patch clamp electrophysiology) with good DMPK properties, acceptable in vivo rat PK and excellent brain levels. Electrophysiology studies in subthalamic nucleus (STN) neurons demonstrated robust effects of ML218 on the inhibition of T-Type calcium current, inhibition of low threshold spike and rebound burst activity. Based on the basal ganglia circuitry in Parkinson's disease (PD), the effects of ML218 in STN neurons suggest a therapeutic role for T-type Ca(2+) channel inhibitors, and ML218 was found to be orally efficacious in haloperidol-induced catalepsy, a preclinical PD model, with comparable efficacy to an A(2A) antagonist, a clinically validated PD target. ML218 proves to be a powerful new probe to study T-Type Ca(2+) function in vitro and in vivo, and freely available.
T型钙通道抑制剂在治疗疼痛、癫痫、睡眠障碍、特发性震颤及其他神经疾病方面具有巨大的治疗潜力;然而,缺乏真正具有选择性的工具阻碍了基础研究,而制药行业的选择性工具可能受到知识产权(IP)限制。因此,开展了一项分子图书馆探针中心(MLPCN)高通量筛选(HTS),以鉴定无IP限制且可通过MLPCN免费获取的新型T型钙通道抑制剂,供生物医学界用于研究T型钙通道。虽然高通量筛选发现了许多活性化合物,但这些化合物无法优化到所需的效价水平以成为合适的工具化合物。因此,在SurflexSim的指导下采用骨架跃迁方法,最终得到了ML218(化学物质标识符45115620),一种选择性T型钙通道(Ca(v)3.1、Ca(v)3.2、Ca(v)3.3)抑制剂(在钙流实验中,Ca(v)3.2的半数抑制浓度(IC(50))=150 nM;在膜片钳电生理实验中,Ca(v)3.2的IC(50)=310 nM,Ca(v)3.3的IC(50)=270 nM),具有良好的药物代谢动力学(DMPK)特性、可接受的大鼠体内药代动力学及出色的脑内浓度。对丘脑底核(STN)神经元的电生理研究表明,ML218对T型钙电流的抑制、低阈值尖峰抑制及反弹爆发活动具有显著作用。基于帕金森病(PD)的基底神经节回路,ML218在STN神经元中的作用表明T型钙通道抑制剂具有治疗作用,且在氟哌啶醇诱导的僵住症(一种临床前PD模型)中发现ML218口服有效,其疗效与临床验证的PD靶点A(2A)拮抗剂相当。ML218被证明是一种强大的新型探针,可用于体外和体内研究T型钙通道功能,且可免费获取。