新藥安全藥理學(xué)研究與進(jìn)展課件

1、新藥安全藥理學(xué)研究與進(jìn)展 第二軍醫(yī)大學(xué)藥物安全性評(píng)價(jià)中心 陸國(guó)才 袁伯俊內(nèi) 容SFDA安全藥理研究要求1ICH安全藥理研究要求2安全藥理研究趨勢(shì)展望3安全藥理學(xué)：研究藥物治療范圍內(nèi)或治療范圍以上的劑量時(shí)，潛在不期望出現(xiàn)的對(duì)生理功能的不良影響次要藥效學(xué)：主要研究藥物非期望的、與治療目的不相關(guān)的效應(yīng)和作用機(jī)制概 念SFDA要求核心組合試驗(yàn)(Core Battery) 中樞神經(jīng)系統(tǒng)、心血管系統(tǒng)和呼吸系統(tǒng)研究追加的安全藥理學(xué)研究（Follow-up） 對(duì)中樞神經(jīng)系統(tǒng)、心血管系統(tǒng)和呼吸系統(tǒng)進(jìn)行深入的研究 補(bǔ)充的安全藥理學(xué)研究（Supplemental） 研究藥物對(duì)三大系統(tǒng)以外器官(如泌尿系統(tǒng)、自主神經(jīng)系

2、統(tǒng)、胃腸道系統(tǒng)和其它器官組織)功能的影響概 念SFDA要求確定藥物可能存在的非期望藥效作用評(píng)價(jià)藥物在毒理學(xué)和/或臨床研究中所觀察到的藥物不良反應(yīng)和/或病理生理作用研究所觀察到的和/或推測(cè)的藥物不良反應(yīng)機(jī)制目 的SFDA要求有助于了解新藥的藥理作用特點(diǎn)和機(jī)制有助于發(fā)現(xiàn)新藥理作用，一藥多用，藥盡其用為毒理學(xué)研究打下基礎(chǔ)，是各種毒性實(shí)驗(yàn)的必要補(bǔ)充目 的SFDA要求貫穿在新藥研究全過(guò)程中，可分階段進(jìn)行在藥物進(jìn)入臨床試驗(yàn)前，應(yīng)完成Core Battery Studies追加和/或補(bǔ)充的安全藥理學(xué)研究可在申報(bào)生產(chǎn)前進(jìn)行階段性SFDA要求內(nèi)容要求SFDA要求神經(jīng)系統(tǒng) 運(yùn)動(dòng)功能、行為改變、協(xié)調(diào)功能、感覺(jué)運(yùn)動(dòng)反

3、射和體溫等的變化(小鼠)心血管系統(tǒng) 給藥前后血壓（包括收縮壓、舒張壓和平均壓）、心電圖（包括QT間期、PR間期、ST段和QRS波等）和心率等變化(Beagle犬)呼吸系統(tǒng) 呼吸頻率和呼吸幅度(Beagle犬)根據(jù)情況增加其它系統(tǒng)或指標(biāo)1.動(dòng)物選擇 可采用不同類型動(dòng)物(正?；蚰Ｐ停逍鸦蚵樽?。一般多用正常動(dòng)物2.劑量途徑 產(chǎn)生主要藥效作用及以上的3個(gè)劑量，臨床給藥途徑3.觀察時(shí)間 藥效學(xué)和藥代動(dòng)力學(xué)特性、受試動(dòng)物、臨床研究方案等因素選擇觀察時(shí)間點(diǎn)和觀察時(shí)間SFDA要求有關(guān)問(wèn)題4.系統(tǒng)指標(biāo)中樞神經(jīng)系統(tǒng)心血管系統(tǒng)呼吸系統(tǒng)根據(jù)情況增加其它系統(tǒng)或指標(biāo)5.體外研究 應(yīng)確定受試物的濃度-效應(yīng)關(guān)系。無(wú)明顯影

4、響作用時(shí)，應(yīng)說(shuō)明濃度選擇范圍SFDA要求有關(guān)問(wèn)題中樞神經(jīng)系統(tǒng)：對(duì)行為、學(xué)習(xí)記憶、神經(jīng)生化、視覺(jué)、聽(tīng)覺(jué)和電生理等的影響心血管系統(tǒng)：對(duì)心輸出量、心肌收縮作用、血管阻力等指標(biāo)的檢測(cè)呼吸系統(tǒng)：對(duì)氣道阻力、肺動(dòng)脈壓力、血?dú)夥治龅戎笜?biāo)的檢測(cè)SFDA要求追加安全藥理學(xué)泌尿系統(tǒng)：對(duì)腎功能的影響，如對(duì)尿量、比重、滲透壓、pH、電解質(zhì)平衡、蛋白質(zhì)、細(xì)胞和血生化（如尿素氮、肌酐、蛋白質(zhì)）等指標(biāo)的檢測(cè)自主神經(jīng)系統(tǒng)：有關(guān)受體的結(jié)合、體內(nèi)或體外對(duì)激動(dòng)劑或拮抗劑的功能反應(yīng)、對(duì)自主神經(jīng)的直接刺激作用和對(duì)心血管反應(yīng)、壓力反射和心率等指標(biāo)的檢測(cè)SFDA要求補(bǔ)充安全藥理學(xué)胃腸系統(tǒng)：觀察藥物對(duì)胃腸系統(tǒng)的影響，如胃液分泌量和pH、胃腸

5、損傷、膽汁分泌、離體回腸收縮、胃液pH等指標(biāo)的測(cè)定其它研究：潛在的依賴性、骨骼肌、免疫和內(nèi)分泌功能等的影響SFDA要求補(bǔ)充安全藥理學(xué)體內(nèi)血藥濃度低，或其它組織器官分布很少的局部用藥（如皮膚、眼科用藥）只用于治療晚期癌癥病人的細(xì)胞毒類藥物，在首次用于臨床前可不做一般藥理學(xué)研究，但不包括具有新作用機(jī)制的此類藥物SFDA要求免做安全藥理學(xué)內(nèi) 容SFDA安全藥理研究要求1ICH安全藥理研究要求2安全藥理研究趨勢(shì)展望3“Safety pharmacology includes the assessment of effects on vital functions, such as cardiovasc

6、ular, central nervous system and respiratory systems, and these should be evaluated prior to human exposure.” ICH M3: Nonclinical safety studies for the conduct of human clinical trials for pharmaceuticals. (July 1997)Value of safety pharmacology acknowledged by regulators and sponsorsICH要求ICH Guide

7、line M3, 1997“Safety pharmacology studies . investigate the potential for undesirable pharmacological activity in appropriate animal models and, where necessary, to incorporate particular monitoring for these activities in the toxicity studies and/or clinical studies.”“. these studies may allow for

8、a mechanistically-based explanation of specific organ toxicities, which should be considered carefully with respect to human use and indication(s).”ICH S6 “Preclinical Safety Evaluation of Biotechnology-Derived Pharmaceuticals” (July 1997)Pharmacological activity in animals should be considered in t

9、oxicology and clinical studiesMechanism-based understanding of toxicities (or pharmacological activities) should be carefully considered in the risk assessmentICH要求ICH Guideline S6, 1997“This guidance was developed to help protect clinical trial participants and patients receiving marketed products

10、from potential adverse effects of pharmaceuticals, while avoiding unnecessary use of animals and other resources.”ICH S7A “Safety Pharmacology Studies for Human Pharmaceuticals” (July 2001)Safety pharmacology studies have the potential to reduce use of animals and other resourcesICH要求ICH Guideline S

11、7A, 2001清醒動(dòng)物-遙測(cè)系統(tǒng)檢測(cè)清醒狀態(tài)下、自由活動(dòng)動(dòng)物的安全藥理學(xué)指標(biāo) 節(jié)省動(dòng)物符合動(dòng)物福利要求適合不同給藥方式連續(xù)動(dòng)態(tài)、長(zhǎng)時(shí)間檢測(cè)指標(biāo)ICH要求Animal IDCompound/Dose PeriodDose Period 1Day 1Dose Period 2Day 4Dose Period 3Day 8Dose Period 4Day 11No.1LHCMNo.2HMLCNo.3CLMHNo.4MCHL4 x 4拉丁方實(shí)驗(yàn)設(shè)計(jì)ICH要求Animal IDCompound/Dose PeriodDose Period 1Day 1Dose Period 2Day 4Dose Pe

12、riod 3Day 8Dose Period 4Day 11No.1LHCMNo.2HMLCNo.3CLMHNo.4MCHLNo.5LHCMNo.6HMLCNo.7CLMHNo.8MCHL4 x 8拉丁方實(shí)驗(yàn)設(shè)計(jì)ICH要求Drug to increase gastrointestinal (GI) motilityOver 5.4 million prescriptions in 1996 (US)34 patients TdP, 23 long QT (1993-1996) Cardiotoxicity reported when used with Ketoconazole and/or

13、ErythromycinCase report of 8-year old girlintensive careICH要求Cisapride StoryICH要求Drug Interactions Leading to QTDrug to increase gastrointestinal (GI) motilityOver 5.4 million prescriptions in 1996 (US)34 patients TdP, 23 long QT 1993-1996 Cardiotoxicity reported when used with Ketoconazole and/or E

14、rythromycinCase report of 8-year old girlintensive care4 deaths and 16 resuscitated from 1993 to 1996Drug withdrawn from US market July, 2000ICH要求Cisapride StoryFDA recommended removal of Terfenadine containing products from the market in 01/1997FDA issued warning letter to Doctors about cardiac saf

15、ety of CisapridePost-marketing surveillance became more important Medwatch reporting systemSafety pharmacology guidances were approved to look at cardiovascular, respiratory and CNS side-effects of drug candidates ICH S7A and S7BScreening for QT prolongation is now an integral part of drug developme

16、nt (lead optimization to clinical)ICH要求Regulatory Response“The study results can be used to elucidate the mechanism of action and, when considered with other information, estimate risk for delayed ventricular repolarization and QT interval prolongation in humans.”ICH S7B: Nonclinical Evaluation of t

17、he Potential for Delayed Ventricular Repolarization (QT Interval Prolongation) by Human Pharmaceuticals (October 2005)Mechanism of action can be a goal of studiesUse data with other data to estimate risk for QT prolongation in humansICH要求ICH Guideline S7B, 2005QTc (msec)MaleFemaleNormal430450470Baze

18、tt: QTc = QT / RR 0.5 Fridericia: QTc = QT / RR 0.33ICH要求QTc Interval心肌動(dòng)作電位時(shí)的離子通道ICH要求IKr: 快速延遲整流K+電流 QT/QTc Prolongation IKr InhibitionDelayed Ventricular RepolarizationArrhythmia ADDITIONAL RISK FACTORSIKr inhibitory potency: can help predict risk of QT/QTc prolongation is a biomarker for increase

19、d risk of Ventricular Arrhythmias is not a biomarker for Torsade de PointesICH要求 IKr &TdPICH要求Early after-depolarization（早后去極化） TdP Drug Class Date WithdrawnTerfenadineAntihistamineFeb 1998SertindoleAntipsychoticDec 1998AstemizoleAntihistamineJun 1999Grepafloxacin AntibioticNov 1999CisaprideGI Proki

20、neticJuly 2000ICH要求Drugs Withdrawn for TdP risk采用離體動(dòng)物或人體心臟細(xì)胞、培養(yǎng)心臟細(xì)胞系或克隆人離子通道的異種表達(dá)體系測(cè)定離子流在離體心臟標(biāo)本進(jìn)行動(dòng)作電位參數(shù)測(cè)定，或在麻醉動(dòng)物中進(jìn)行能體現(xiàn)動(dòng)作電位周期的特異性電生理參數(shù)檢測(cè)測(cè)定清醒或麻醉動(dòng)物的ECG參數(shù)在離體心臟標(biāo)本或動(dòng)物進(jìn)行致心律失常作用測(cè)定QT間期延長(zhǎng)非臨床研究方法ICH要求內(nèi) 容SFDA安全藥理研究要求1ICH安全藥理研究要求2安全藥理研究趨勢(shì)展望32009趨勢(shì)展望安全藥理學(xué)政策法規(guī)不斷完善趨勢(shì)展望QT間期研究策略體外電生理IKr研究采用原代或表達(dá)的IKr通道蛋白(如由hERG編碼的蛋白)評(píng)

21、價(jià)藥物對(duì)離子電流的影響體內(nèi)對(duì)QT的研究從體內(nèi)測(cè)定心室復(fù)極化，如QT間期檢測(cè)結(jié)果進(jìn)行評(píng)價(jià)化學(xué)/藥理的分類可誘導(dǎo)人QT間期延長(zhǎng)作用的藥物如抗精神病類藥物，組胺H-1受體拮抗劑、氟喹酮類QT間期研究策略趨勢(shì)展望追加試驗(yàn)離體心臟 麻醉動(dòng)物 重復(fù)給藥試驗(yàn)動(dòng)物種屬和性別的選擇用代謝性誘導(dǎo)劑或用拮抗劑用陽(yáng)性藥和參比化合物其他通道的抑制作用在多個(gè)時(shí)間點(diǎn)測(cè)定電生理參數(shù)模擬病理狀況和心律失常綜合風(fēng)險(xiǎn)評(píng)估 QT間期研究策略趨勢(shì)展望計(jì)算機(jī)模擬 (in silico)干細(xì)胞定向分化為心肌細(xì)胞下丘腦病理切片HERG通道體外篩選新方法熒光篩選(測(cè)量膜電位)結(jié)合試驗(yàn) (標(biāo)記的HERG通道阻滯劑，HTS)斑馬魚HTS結(jié)合藥代/

22、藥效，研究毒理學(xué)生物標(biāo)志，預(yù)測(cè)臨床安全藥理學(xué)研究新技術(shù)新方法趨勢(shì)展望計(jì)算機(jī)模擬 (in silico)干細(xì)胞定向分化為心肌細(xì)胞下丘腦病理切片HERG通道體外篩選新方法熒光篩選(測(cè)量膜電位)結(jié)合試驗(yàn) (標(biāo)記的HERG通道阻滯劑，HTS)斑馬魚HTS結(jié)合藥代/藥效，研究毒理學(xué)生物標(biāo)志，預(yù)測(cè)臨床安全藥理學(xué)研究新技術(shù)新方法趨勢(shì)展望PART I MUTAGENESIS AND MUTANTSHighly Efficient ENU Mutagenesis in ZebrafishProduction of Pseudotyped Retrovirus and the Generation of Prov

23、iral Transgenic ZebrafishUndertaking a Successful Gynogenetic Haploid Screen in ZebrafishCryopreservation and In Vitro Fertilization at the Zebrafish International Resource Center計(jì)算機(jī)模擬 (in silico)干細(xì)胞定向分化為心肌細(xì)胞下丘腦病理切片HERG通道體外篩選新方法熒光篩選(測(cè)量膜電位)結(jié)合試驗(yàn) (標(biāo)記的HERG通道阻滯劑，HTS)斑馬魚HTS結(jié)合藥代/藥效，研究毒理學(xué)生物標(biāo)志，預(yù)測(cè)臨床安全藥理學(xué)研究新技術(shù)

24、新方法趨勢(shì)展望PART II TRANSGENESISTransient and Stable Transgenesis Using Tol2 Transposon VectorsAnalysis of Genes and Genome by the Tol2-Mediated Gene and Enhancer Trap MethodsBacterial Artificial Chromosome Transgenesis for ZebrafishSimple and Efficient Transgenesis with Meganuclease Constructs in Zebraf

25、ish計(jì)算機(jī)模擬 (in silico)干細(xì)胞定向分化為心肌細(xì)胞下丘腦病理切片HERG通道體外篩選新方法熒光篩選(測(cè)量膜電位)結(jié)合試驗(yàn) (標(biāo)記的HERG通道阻滯劑，HTS)斑馬魚HTS結(jié)合藥代/藥效，研究毒理學(xué)生物標(biāo)志，預(yù)測(cè)臨床安全藥理學(xué)研究新技術(shù)新方法趨勢(shì)展望PART III TISSUE-SPECIFIC MANIPULATIONSNitroreductase-Mediated Cell Ablation in Transgenic Zebrafish EmbryosFocal Electroporation in Zebrafish Embryos and LarvaeTissue Mi

26、cromanipulation in Zebrafish Embryos計(jì)算機(jī)模擬 (in silico)干細(xì)胞定向分化為心肌細(xì)胞下丘腦病理切片HERG通道體外篩選新方法熒光篩選(測(cè)量膜電位)結(jié)合試驗(yàn) (標(biāo)記的HERG通道阻滯劑，HTS)斑馬魚HTS結(jié)合藥代/藥效，研究毒理學(xué)生物標(biāo)志，預(yù)測(cè)臨床安全藥理學(xué)研究新技術(shù)新方法趨勢(shì)展望PART IV ANALYZING GENE EXPRESSIONZebrafish Spotted-Microarray for Genome-Wide Expression Profiling Experiments. Part I: Array Printing a

27、nd HybridizationPart II : Data Acquisition and AnalysisValidating microRNA Target Transcripts Using Zebrafish Assays計(jì)算機(jī)模擬 (in silico)干細(xì)胞定向分化為心肌細(xì)胞下丘腦病理切片HERG通道體外篩選新方法熒光篩選(測(cè)量膜電位)結(jié)合試驗(yàn) (標(biāo)記的HERG通道阻滯劑，HTS)斑馬魚HTS結(jié)合藥代/藥效，研究毒理學(xué)生物標(biāo)志，預(yù)測(cè)臨床安全藥理學(xué)研究新技術(shù)新方法趨勢(shì)展望PART V IMAGINGMultiple Embryo Time-Lapse Imaging of Zebrafish Development Live Cell Imaging of Zebrafish LeukocytesImaging Zebrafish Embryos by Two-Photon Excitation Time-Lapse MicroscopyMorphological Analysis of the Zebrafish Diges