摘要
目的:建立靶向毒蕈碱乙酰胆碱4(ChRM4,M4)调节剂的体内外药效学评价体系,并开展Emraclidine抗精神分裂症的体内外药效评价,为M4受体调节剂(PAM)的临床前体内外药效学评价体系提供参考。方法:利用CHO细胞通过Flp-In^(TM)定点整合技术构建可同时稳定整合表达M4和Gqi5嵌合型G蛋白的CHO-Gqi5-M4细胞模型,通过检测胞内钙离子水平来评价Emraclidine对M4受体的正向调节活性;采用地佐环平(MK-801)急性给药诱导C57BL/6J小鼠建立精神分裂症模型,通过前脉冲抑制试验和旷场试验评价Emraclidine的抗精神分裂症作用。结果:在细胞水平,Emraclidine可通过激动M4受体抑制cAMP,从而剂量依赖性降低胞内钙离子水平;在动物实验中,Emraclidine能够缓解MK-801诱导的前脉冲抑制降低和焦虑样行为。结论:采用CHO-Gqi5-M4细胞模型及PPI、旷场等行为学方法,构建了靶向M4调节剂的体内外药效学评价流程,证实Emraclidine可激动ChRM4受体活性和缓解MK-801诱导的精神分裂症状,为ChRM4受体调节剂的临床前体内外药效学评价体系提供参考。
Objective:To evaluate muscarinic acetylcholine receptor M4(ChRM4/M4)modulators,we developed novel pharmacodynamic evaluation systems for both in vitro and in vivo settings.Subsequently,we performed a comprehensive assessment of Emraclidine's anti-schizophrenia efficacy using these established systems.The findings of this study inform the development of a robust preclinical pharmacodynamic evaluation framework for ChRM4 receptor positive allosteric modulators(PAMs).Methods:We used Flp-In^(TM)site-specific integration technology to construct a CHO cell model(CHO-Gqi5-M4)capable of stably co-expressing M4 receptors and chimeric Gqi5 proteins.Intracellular calcium levels were measured to evaluate the agonistic activity of Emraclidine on M4 receptors.Additionally,an acute dizocilpine(MK-801)administration regimen was employed to induce a schizophrenia-like phenotype in C57BL/6J mice.The anti-schizophrenia effects of Emraclidine were subsequently assessed through prepulse inhibition(PPI)and open-field tests.Results:At the cellular level,Emraclidine inhibits cAMP through M4 receptor agonism,thereby dose-dependently reducing intracellular calcium levels.In animal studies,it ameliorated MK-801-induced deficits in prepulse inhibition(PPI)and anxiety-like behaviors.Conclusion:Using the CHO-Gqi5-M4 cell model and behavioral methods including PPI and open field test,we established an in vitro and in vivo pharmacodynamic evaluation workflow for M4-targeted modulators.Using this system,we demonstrated that Emraclidine effectively activates ChRM4 receptor signaling and alleviates MK-801-induced schizophrenia-like behaviors.These findings establish a valuable framework for the preclinical pharmacodynamic evaluation of ChRM4-targeting therapeutics.
作者
黄晓凤
苏明波
王立辉
HUANG Xiaofeng;SU Mingbo;WANG Lihui(School of Clinical Pharmacy,Shenyang Pharmaceutical University,Shenyang 110000,China;Yangtze Delta Drug Advanced Research Institute,Nantong 226133,China;Nantong Hongye Pharmaceutical Company,Nantong 226133,China)
出处
《甘肃医药》
2026年第2期162-166,共5页
Gansu Medical Journal
