Frötschl Roland
BfArM-Bundesinstitut für Arzneimittel und Medizinprodukte, Federal Institute for Drugs and Medical Devices, Kurt-Georg-Kiesinger-Allee 3, 53175 Bonn, Germany
Mutagenesis. 2015 Jan;30(1):51-7. doi: 10.1093/mutage/geu069.
The in vivo comet assay has recently been implemented into regulatory genotoxicity testing of pharmaceuticals with inclusion into the ICH S2R1 guidance. Regulatory genotoxicity testing aims to detect DNA alterations in form of gene mutations, larger scale chromosomal damage and recombination and aneuploidy. The ICH S2R1 guideline offers two options of standard batteries of tests for the detection of these endpoints. Both options start with an AMES assay and option 1 includes an in vitro mammalian cell assay and an in vivo micronucleus assay in rodent, whereas option 2 includes an in vivo micronucleus assay in bone marrow in rodent and a second in vivo assay in a second tissue with a second endpoint. The test recommended as second in vivo test is the comet assay in rat liver. The in vivo comet assay is considered as mature enough to ensure reliable detection of relevant in vivo genotoxicants in combination with the micronucleus test in bone marrow and the AMES assay. Although lots of research papers have been published using the in vitro comet assay, the in vitro version has not been implemented into official regulatory testing guidelines. A survey of the years 1999-2014 revealed 27 in vivo comet assays submitted to BfArM with market authorisation procedures, European and national advice procedures and clinical trial applications. In three procedures, in vitro comet assays had been submitted within the genetic toxicology packages.
体内彗星试验最近已被纳入药品监管遗传毒性测试，并写入了国际人用药品注册技术协调会（ICH）的S2R1指南。监管遗传毒性测试旨在检测基因突变、大规模染色体损伤、重组和非整倍体等形式的DNA改变。ICH S2R1指南提供了两种标准测试组合选项，用于检测这些终点。两种选项均以艾姆斯试验开始，选项1包括体外哺乳动物细胞试验和啮齿动物体内微核试验，而选项2包括啮齿动物骨髓体内微核试验和第二个组织中针对第二个终点的第二个体内试验。推荐作为第二个体内试验的是大鼠肝脏彗星试验。体内彗星试验被认为已经足够成熟，能够与骨髓微核试验和艾姆斯试验相结合，可靠地检测相关的体内遗传毒性物质。尽管已经发表了许多使用体外彗星试验的研究论文，但体外版本尚未被纳入官方监管测试指南。一项对1999年至2014年的调查显示，有27项体内彗星试验提交给了德国联邦药品和医疗器械研究所（BfArM），用于市场授权程序、欧洲和国家咨询程序以及临床试验申请。在三个程序中，体外彗星试验已在遗传毒理学包中提交。
