Xanomeline新型衍生物SBG-PK-014促进APPsw的α-剪切

华东师范大学学报(自然科学版) ›› 2014, Vol. 2014 ›› Issue (1): 123-132.

Xanomeline新型衍生物SBG-PK-014促进APPsw的α-剪切

王栋¹，周宗丽¹，高虹¹，雷小平²，董素珍¹，胡金锋^1,3

1. 华东师范大学脑功能基因组学教育部重点实验室, 上海 200062；
2. 北京大学药学院, 北京 100191； 3. 复旦大学药学院, 上海 201203

收稿日期:2013-04-01 修回日期:2013-07-01 出版日期:2014-01-25 发布日期:2015-09-25

Novel derivative of xanomeline, SBG-PK-014, increases the α-secretion of APPsw

WANG Dong¹, ZHOU Zong-li¹, GAO Hong¹, LEI Xiao-ping²,DONG Su-zhen¹, HU Jin-feng^1,3

1. Key Laboratory of Brain Functional Genomics, Ministry of Education, East China Normal University, Shanghai 200062, China; 2. School of Pharmaceutical Sciences, Peking University, Beijing 100191, China; 
3. School of Pharmacy, Fudan University, Shanghai 201203, China

Received:2013-04-01 Revised:2013-07-01 Online:2014-01-25 Published:2015-09-25

摘要/Abstract

摘要： 研究了xanomeline新型衍生物SBG-PK-014对毒蕈碱型M1乙酰胆碱受体的激活能力以及对APP基因瑞典型突变体(APPsw)的α-剪切的作用.利用M1激动剂筛选细胞模型检测了SBG-PK-014的EC₅₀和最大响应倍数(FA_max)，并在小鼠神经母细胞瘤N2a细胞中同时过表达APPsw和M1受体，分析了该化合物和xanomeline对sAPPα分泌的影响.结果显示，SBG-PK-014的EC₅₀(40.2 nmol/L)与xanomeline (28.4 nmol/L)接近，但FA_max是xanomeline的3.5倍.SBG-PK-014通过激活M1受体促进APPsw的α-剪切，且在0.1 μmol/L和1 μmol/L的浓度下，其效果显著强于同剂量的xanomeline.可见，SBG-PK-014比xanomeline更能有效地激活M1受体，还能促进APPsw的α-剪切和神经保护性sAPPα的生成，在调节阿尔茨海默病的Aβ病理途径上可能有一定潜力，值得进一步研究.

关键词: xanomeline, SBG-PK-014, M1受体激动剂, 阿尔茨海默病, 衰老

Abstract: The activity of a novel derivative of xanomeline, SBG-PK-014, on muscarinic M1 mAChRs and the α-secretion of human APP Swedish mutant (APPsw) was evaluated. The EC₅₀ and maximum folds of activation (FA_max) were measured in a cell-based model. Mouse N2a cells over-expressing both APPsw and M1 mAChR gene were treated with SBG-PK-014 and xanomeline, respectively. Their secreation levels of sAPPα were then measured using Western Blotting. The results showed that SBG-PK-014 had a similar EC₅₀ to xanomeline (40.2 nmol/L vs. 28.4 nmol/L), but demonstrated a 3.5-fold FA_max, as compared to xanomeline. SBG-PK-014 promoted the α-secretion of APPsw via the activation of M1 receptors. At the same dose of 0.1 μmol/L and 1 μmol/L, SBG-PK-014 exhibited significantly more potent activity. SBG-PK-014 activated M1 receptors more effectively than xanomeline, increased the α-cut of APPsw as well as the secretion of neuroprotective sAPPα, showed potential in modifying the Aβ pathology of Alzheimer’s disease (AD), and is worth further development.

Key words: xanomeline, SBG-PK-014, M1 agonist, Alzheimer’s disease, aging

中图分类号:

Q291

王栋，周宗丽，高虹，雷小平，董素珍，胡金锋. Xanomeline新型衍生物SBG-PK-014促进APPsw的α-剪切[J]. 华东师范大学学报(自然科学版), 2014, 2014(1): 123-132.

WANG Dong, ZHOU Zong-li, GAO Hong, LEI Xiao-ping,DONG Su-zhen, HU Jin-feng. Novel derivative of xanomeline, SBG-PK-014, increases the α-secretion of APPsw[J]. Journal of East China Normal University(Natural Sc, 2014, 2014(1): 123-132.

参考文献

［1］ CONTESTABILE A. The history of the cholinergic hypothesis［J］. Behav Brain Res,2011, 221(2):334-340.

［2］ KARRAN E, MERCKEN M, DE STROOPER B. The amyloid cascade hypothesis for Alzheimer's disease: an appraisal for the development of therapeutics［J］. Nat Rev Drug Discov,2011, 10(9):698-712.

［3］ CHASSEIGNEAUX S, ALLINQUANT B. Functions of Abeta, sAPPalpha and sAPPbeta : similarities and differences［J］. J Neurochem,2012, 120 Suppl 1:99-108.

［4］ WEI J, WALTON EA, MILICI A, et al. m1-m5 muscarinic receptor distribution in rat CNS by RT-PCR and HPLC［J］. J Neurochem,1994, 63(3):815-821.

［5］ CACCAMO A, ODDO S, BILLINGS LM, et al. M1 receptors play a central role in modulating AD-like pathology in transgenic mice［J］. Neuron,2006, 49(5):671-682.

［6］ FISHER A. Cholinergic modulation of amyloid precursor protein processing with emphasis on M1 muscarinic receptor: perspectives and challenges in treatment of Alzheimer's disease［J］. J Neurochem,2012, 120 Suppl 1:22-33.

［7］ SVENSSON AL, ALAFUZOFF I, NORDBERG A. Characterization of muscarinic receptor subtypes in Alzheimer and control brain cortices by selective muscarinic antagonists［J］. Brain Res,1992, 596(1-2):142-148.

［8］ BODICK NC, OFFEN WW, LEVEY AI, et al. Effects of xanomeline, a selective muscarinic receptor agonist, on cognitive function and behavioral symptoms in Alzheimer disease［J］. Arch Neurol,1997, 54(4):465-473.

［9］高虹, 欧阳克清, 郑旭煦, 等. 毒蕈胆碱M1受体激动剂的高通量筛选模型［J］. 中国药理学通报,2003, 19(7):776-779.

［10］ ODDO S, CACCAMO A, SHEPHERD JD, et al. Triple-transgenic model of Alzheimer's disease with plaques and tangles: intracellular Abeta and synaptic dysfunction［J］. Neuron,2003, 39(3):409-421.

［11］ CHEN CD, PODVIN S, GILLESPIE E, et al. Insulin stimulates the cleavage and release of the extracellular domain of Klotho by ADAM10 and ADAM17［J］. Proc Natl Acad Sci U S A,2007, 104(50):19796-19801.

［12］ FU H, DOU J, LI W, et al. Promising multifunctional anti-Alzheimer's dimer bis(7)-Cognitin acting as an activator of protein kinase C regulates activities of alpha-secretase and BACE-1 concurrently［J］. Eur J Pharmacol,2009, 623(1-3):14-21.

［13］ CAILLE I, ALLINQUANT B, DUPONT E, et al. Soluble form of amyloid precursor protein regulates proliferation of progenitors in the adult subventricular zone［J］. Development,2004, 131(9):2173-2181.

［14］ GAKHAR-KOPPOLE N, HUNDESHAGEN P, MANDL C, et al. Activity requires soluble amyloid precursor protein alpha to promote neurite outgrowth in neural stem cell-derived neurons via activation of the MAPK pathway［J］. Eur J Neurosci,2008, 28(5):871-882.

［15］ FREUDE KK, PENJWINI M, DAVIS JL, et al. Soluble amyloid precursor protein induces rapid neural differentiation of human embryonic stem cells［J］. J Biol Chem,2011, 286(27):24264-24274.

［16］ TAYLOR CJ, IRELAND DR, BALLAGH I, et al. Endogenous secreted amyloid precursor protein-alpha regulates hippocampal NMDA receptor function, long-term potentiation and spatial memory［J］. Neurobiol Dis,2008, 31(2):250-260.

［17］ DAVIS AA, FRITZ JJ, WESS J, et al. Deletion of M1 muscarinic acetylcholine receptors increases amyloid pathology in vitro and in vivo［J］. J Neurosci,2010, 30(12):4190-4196.

［18］ CACCAMO A, FISHER A, LAFERLA FM. M1 agonists as a potential disease-modifying therapy for Alzheimer's disease［J］. Curr Alzheimer Res,2009, 6(2):112-117.

［19］ ECKOLS K, BYMASTER FP, MITCH CH, et al. The muscarinic M1 agonist xanomeline increases soluble amyloid precursor protein release from Chinese hamster ovary-m1 cells［J］. Life Sci,1995, 57(12):1183-1190.

［20］ DELAPP N, WU S, BELAGAJE R, et al. Effects of the M1 agonist xanomeline on processing of human beta-amyloid precursor protein (FAD, Swedish mutant) transfected into Chinese hamster ovary-m1 cells［J］. Biochem Biophys Res Commun,1998, 244(1):156-160.

［21］ BYMASTER FP, CARTER PA, PETERS SC, et al. Xanomeline compared to other muscarinic agents on stimulation of phosphoinositide hydrolysis in vivo and other cholinomimetic effects［J］. Brain Res,1998, 795(1-2):179-190.

［22］ WOOD MD, MURKITT KL, HO M, et al. Functional comparison of muscarinic partial agonists at muscarinic receptor subtypes hM1, hM2, hM3, hM4 and hM5 using microphysiometry［J］. Br J Pharmacol,1999, 126(7):1620-1624.

［23］ SAMS AG, HENTZER M, MIKKELSEN GK, et al. Discovery of N-{1-［3-(3-oxo-2,3-dihydrobenzo［1,4］oxazin-4-yl)propyl］piperidin-4-yl}-2-p henylacetamide (Lu AE51090): an allosteric muscarinic M1 receptor agonist with unprecedented selectivity and procognitive potential［J］. J Med Chem,2010, 53(17):6386-6397.

［24］ VEROFF AE, BODICK NC, OFFEN WW, et al. Efficacy of xanomeline in Alzheimer disease: cognitive improvement measured using the Computerized Neuropsychological Test Battery (CNTB)［J］. Alzheimer Dis Assoc Disord,1998, 12(4):304-312.

[1]	张盼, 俞彬, 刘莎, 段雅乐, 赵政. 姜黄素对老年鼠脑内氧化性平衡和星形胶质细胞的影响[J]. 华东师范大学学报(自然科学版), 2015, 2015(6): 108-116.
[2]	王栋，翟文珠，苏静静，孟博，赵政，胡金锋. 早老素PS1/PS2双敲除小鼠中klotho表达的下降和血磷的升高[J]. 华东师范大学学报(自然科学版), 2013, 2013(2): 75-83, 91.
[3]	张双，王琳，董素珍. 利用基因芯片检测大鼠衰老相关的记忆障碍的调控基因[J]. 华东师范大学学报(自然科学版), 2011, 2011(6): 89-99.
[4]	司文;白静;邓世宁;曾庆文;曹晓华. 镁对老年小鼠学习和记忆的改善作用[J]. 华东师范大学学报(自然科学版), 2008, 2008(6): 96-102.