Investigate all ion channels -Instead of only a few
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1 Investigate all ion channels -Instead of only a few ICE Bioscience, Inc Weimin Pei, Chief Science officer peiwm@ice-biosci.com 1
2 About me 16 years research on ion channel in US 7 Years in commercial service 9 years GLP and other safety experience Work with almost all the big Pharm globally Touch almost all the ion channels
3 All ion channels His skill and insight in cardiomyocyte and recombinant cell lines have been critically important to the success of ChanTest as contract research organization Weimin worked on almost all the ion channel services in ChanTest, and understands ion channel function and pharmacology As a study director for both manual and automated patch studies Weimin has proven himself to be one of the most talented electrophysiologist 他对离子通道重组细胞系列的深刻理解及技能对于 ChanTest 的成功至关重要.. 作为手动和自动电生理平台的研究总监 证明为最具才华的电生理学家之一 Glenn E. Kirsch PhD, Case Western Professor, ChanTest Tech VP served as Senior Scientist/Study Director for Ion Channel Weimin frequently served an essential function of characterizing the electrophysiology and pharmacology of the new cll lines I found him very knowledgeable about multiple classes of ion channels he taught me a great deal about the intricacies of ion channel function 作为高级研究员 / 研究总监 经常起到极其重要不可缺的作用在新的离子通道细胞系方面 Director Molecular and cell Biology of Charles River Lab. Luke C. Armstrong PhD
4 Our team Dr. Li Yan 闫励博士董事长 (CEO) 德国鲁尔波鸿大学神经生理博士擅长神经电生理学, 药理学研究北京市海外高层次 海聚工程 领军人才, 北京市特聘专家 Dr. Yingji Li 李英骥博士总经理 (General Manager) 德国吉森大学医学博士, 吉林大学生理学博士擅长心血管系统药理学研究北京市海外高层次 海聚工程 领军人才, 北京市特聘专家 Dr. Weimin Pei 博士首席科学官 (CSO) New York State University 博士曾任职 ChanTest(Charles River Laboratory) 对一百多种离子通道的检测具有丰富的经验 与几乎所有知名药企进行过合作北京市海外高层次 海聚工程 领军人才, 北京市特聘专家
5 Our team Dr. Ling Gu 博士 Medical Engineering 浙江大学生物医学工程博士美国 University of Texas 博士后曾在 LaiTag Science 公司担任 CSO Dr. Xu Zhang 张旭博士 Electrophysiology China Science Institute 中科院生物物理所分子生物学博士曾担任康龙化成电生理部长 Dr. Zhaoli Guan 关丽照博士 Molecular and Cell Biology Beijing University 北京大学医学博士 Dr. Xiaowei Wang 博士 Neuroscience Biology Brisbane University 医学博士
6 Our team Dr. Ling Gu 博士市场总监浙江大学生物医学工程博士美国 University of Texas 博士后曾在 LaiTag Science 公司担任 CSO Dr. Xu Zhang 张旭博士技术总监 China Science Institute 中科院生物物理所分子生物学博士曾担任康龙化成电生理部长 Dr. Zhaoli Guan 关丽照博士分子生物学总监 Beijing University 北京大学医学博士 Dr. Xiaowei Wang 博士神经生物学总监 Beijing University 北京大学医学博士
7 What makes ICE special Young hard working Scientist team(7 PhD, 8 MS, and 15 BS) China s best ion channel CRO company with great potential Focus on ion channel and relevant neuroscience The largest ion channel library in China
8 Our Goal: All Ion Channels The largest commercial ion channel CRO in the world With the largest ion channel library (all ion channels available) Platforms of all relevant primary cell, tissue, in vitro, in vivo Science oriented, not money oriented
9 Part of Ion Channel Library Na channel family Ca channel family K channel family Ligand gated channel TRP channel Na channel family Ca channel family K channel family Ligand gated channel Na channel family Ca channel family K channel family Ligand gated channel TRP channel Na channel family K channel family Ligand gated channel Pain NaV1.1, NaV1.2, NaV1.3, NaV1.7, Nav1.8, NaV1.9 CaV2.1, CaV2.2, CaV3.2 KV1.3, KV1.4, KV4.3/KChIP2.2, KV7.2/KV7.3, KCa3.1 GABAA3, NMDA (NR2A,NR2B), P2X3, P2X7 TRPA, TRPV1, TRPV4, TRPM8 Epilepsy NaV1.1, NaV1.2, NaV1.3, NaV1.6 CaV2.1, CaV3.2 KV4.2/KChIP2.2, KV7.2/KV7.3,KV7.3/KV7.5, KCa2.2, KCa2.3 GABA (A1,A2), NMDA (NR2A,NR2B) Anxiety, Depression Related NaV1.1, NaV1.2, NaV1.3, NaV1.7, Nav1.8, NaV1.9 CaV2.1, CaV2.2, CaV3.2 KV7.2/KV7.3,KV7.3/KV7.5,KCa2.1, KCa2.2, KCa2.3 GABA(A2-A5), NMDA (NR2A,NR2B,NR2C), 5HT-3, AchR TRPA, TRPV1, TRPV4, TRPM8 Degenerative Diseases NaV1.1, NaV1.2, NaV1.6 KV1.1, KV1.2, KV1.3, KV1.5, KV3.4, KV7.2/KV7.3,KCa2.1, KCa2.2, KCa2.3 NMDA (NR2A,NR2B,NR2C,NR2D) 9
10 Why All Have to, if want to do science of ion channel For example, in complex system like DRG (Nav1.7) and CM (Cav1.2) Even for blank cell like herg (Asic, Nav) and CHO (TRP) Even cardiac Safety, regulator decide on paradigm shift from one herg to CiPA
11 MICE 60% new molecular entities abandoned due to herg blockade herg blockade alone not specific to predict TdP MICE Matters Because
12 Channels work together From Nature Rev. 2016
13 Consider all cardiac ion channel herg blockade as red flag False Positive False Negative herg Nav1.5, early Nav1.5, late Cav1.2, tonic Cav1.2, late Cav3.2 HCN4 Kv1.5 Kv4.3 KvLQT1/ mink Kir2.1 Terfenadine **** Cisapride **** E-4031**** Vardenafil Pentobarbital Verapamil Vanoxerine
14 25 nm 组织建立了世界上最大的离子 简单介绍离子通道 通道集合库 400 more ion channels <200 commercial Abundant in body Heart & Brain 从电生理学观点 : 人体是个超级复杂的电路网络, 离子通道是其通路开关控制
15 Thank you! 15
16 Torsades de Points (TdP) TdP is a specific type of abnormal heart rhythm that can lead to sudden cardiac death. Main cause of TdP: Congenital (Generic abnormality) Medication - K + blockage Human ether-à-go-go gene (herg) encodes voltage-gated potassium channel Kv11.1, which carries I Kr. 16
17 17
18 Progress of FDA Guidelines on Cardiac safety 2005 ICH S7B herg blockage 2006 ICH E14 Thorough QT test (TQT) 18
19 Are S7B & E14 perfect for Cardiac safety evaluation? herg block QT porlongation TdP 1,False positive results on herg blockage Strong herg blockage, but normal action potential and QT interval. Example: verapamil (CaV1.2), ranolazine (late-na) as many as 60% of new molecular entities developed as potential therapeutic agents, when assayed for herg blocking liability, test positive and are thus abandoned early in development. --De Ponti 2,False negative results on herg blockage No obvious herg blockage, but may cause TdP Among 55 known drugs, 6 have false negative effect and 8 have false positive effect (Kramer J, PNAS 2013) Ikr blockade alone is not highly specific for predicting either delayed repolarization or clinical proarrhythmia 19
20 Are S7B & E14 perfect for Cardiac safety evaluation? 3. S7B allows flexible procedures for testing herg results of the same drug differs a lot from one testing company to another herg results of the same drug differs from one patch clamp system to another herg results of the same drug differs from industry to academia due to the lack of a standard protocol Establish a standard procedure for IC50 test to collect consistent data 20
21 Are S7B & E14 perfect for Cardiac safety evaluation? Very Expensive TQT test , 300 TQT test were reported to FDA TQT were studied in estimation - Each study costed several Million USD The total cost for TQT studies is over Billion USD for the past 10 years 21
22 A New Standard for Cardiac Safety Evaluation Comprehensive in Vitro Proarrhythmia Assay (CiPA) Shift from delayed repolarization to proarrhythmogenicity; replace TQT, revise S7B From Nature Rev
23 Progress of FDA Guidelines on Cardiac safety 2005 ICH S7B herg blockage 2006 ICH E14 Thorough QT test (TQT) 2016/1 Concentration-QTc Use clinical Phase I ECG data to replace TQT 2017/12 Comprehensive in Vitro Proarrhythmia Assay (CiPA) Multi Ion Channel Evaluation (MICE) 23
24 Examples of CiPA -Effect of Verapamil on Cardiac Ion Channels I(pA) Pre- Verapamil 10nM Verapamil 100nM Verapamil 10µM I[nA] Pre- Verapamil-0.3µM Verapamil-1µM Verapamil-3µM Verapamil-10µM Verapamil-30µM time(s) Time[s] Cells: Induced Pluripotent Stem cells (ipsc) I[nA] Pre- Verapamil-3µM Verapamil-10µM Verapamil-30µM Verapamil-100µM Verapamil-300µM herg IC50=2.3μM Cav1.2 IC50=7.1μM Time[s] Nav1.5 IC50=2.3μM 24
25 Effect of Verapamil on Cardiomyocyte Action Potential 1.0 APD90 mv 50 0 control verapamil 10µM Relative to Control Control 0.1M 0.3M 1M 3M 10M APD s Relative to Control Control 0.1M 0.3M 1M 3M 10M 25
26 Late Na + current eliminates herg effect Ranolazine Dofetilide Lidocaine Modified from David G. Strauss, FDA 26
27 CaV1.2 and Nav1.5 vs. herg 27
28 Insilico Simulation of human ventricular electrophysiology 28
29 Induced Pluripotent Stem cell models for cardiomyocytes Advantages of using ipsc: The use of human source stem cells will reduce animal use, and difference between species Unlimited cells Easy to build disease models 29
30 Example II: Effect of Compound X on Cardiac Ion Channels control 0.2uM 1uM 5uM 25uM herg IC50=0.908μM I(nA) I Ks IC50=0.368 μm Cav1.2 IC50=5.64 μm time(s)
31 Effect of Compound X on Cardiac Ion Channels Acute enzyme isolated guinea pig ventricular myocytes Intracellular recording from Purkinje fiber control DRN 25M mv 0-20 control DRN 25µM V (mv) s ipscs derived ventricular myocytes control DRN 10M time(ms) control DRN 25M 20 0 V(mV) 0 V(mV) time(s) time(s) 31
32 Effect of Compound X on Cardiac Ion Channels control mv s E4031 1μM mv s E4031+ 化合物 X mv s
33 CiPA platform ICE complete cardiac panel Recombination HEK293 cell line NaV1.5,CaV1.2,hERG,IKs, Kv1.5,Kir2.1 ipscs derived cardiomyocytes (Patch clamp/ MEA/ Calcium/ contraction) Primary human cardiomyocytes 33
34 Thank you! 34
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