EMEA基因毒性杂质限度指南

1、.20060628 EMEA/CH MP/QWP/251344/2006基因毒性杂质限度指南（中英文对照）Lon do n, 28 June 2006CPMP/SWP /5199/02EMEA/CH MP/QWP /251344/2006The Euro pean Age ncy for the Evaluatio n of Medici nal P roducts欧洲共同体药物评审委员会(EMEA)COMMITTEE FOR MEDICINAL P RODUCTS FOR HUMAN USE人用药品委员会(CHMP)GUIDLINE ON THE LIMITS OF GENOTOXIC IM

2、P URITIES基因毒性杂质限度指南DESCUSSION IN THE SAFETY WORKINGJune 2002-October 2002P ARTY安全工作组之内的讨论TRANSMISSION TO CPMPDecember 2002CPMP传递RELEASE FOR CONSULTATIONDecember 2002专家讨论DEADLINE FOR COMMENTSMarch 2003建议收集最后期限DISCUSSION IN THE SAFETY WORKINGJune 2003-February 2004P ARTY AND QUALITY WORKING P ARTY安全工作

3、组和质量工作组之间的讨论TRANSMISSION TO CPMPMarch 2004转移给CPMP1RE-RELEASE FOR CONSULTATIONJune 2004再次放行给顾问团DEADLINE FOR COMMENTS收集意见的最后期限DISCUSSION IN THE SAFETY WORKINGP ARTY AND QUALITY WORKING P ARTY安全工作组和质量工作组之间的讨论December 2004February 2005-May 2006ADOP TION BY CHMP被CHMP采用28 June 2006DATE FOR COMING INTO EFF

4、ECT生效日期01Ja nuary 2007KEYWORDS关键词Impurities; Genotoxicity; Threshold of toxicological concern (TTC); Structure activity relatio nship (SAR)GUIDLINE ON THE LIMITS OF GENOTOXIC IMP URITIES基因毒性杂质限度指南TABLE OF CONTENTS 目录EXECUTIVE SUMMARY 内容摘要.INTRODUCTION 介绍2. SCOPE 范围3. LEGAL BASIS法律依据4. TOXICOLOGICAL

5、BACKGROUND 毒理学背景5. RECOMMENDATIONS 建议5.1 Geno toxic Compounds With Sufficie nt Evide nee for a Threshold-Related Mecha nism具有充分证据证明其阈值相关机理的基因毒性化合物5.2 Geno toxic Compounds Without Sufficie nt Evide nee for a Threshold-Related Mecha nism不具备充分证据支持其阈值相关机理的基因毒性化合物5.2.1 P harmaceutical Assessment 学评价5.2.2

6、 Toxicological Assessme n毒理学评价5.2.3 App lication of a Threshold of Toxicological C oncern 毒理学担忧阈值应用5.3 Decisi on Tree for Assessme nt of Acce ptability of Geno toxic Imp urities基因毒性杂质可接受性评价决策树REFERENCES.参考文献EXECUTIVE SUMMARY 内容摘要The toxicological assessme nt of geno toxic imp urities and the determ

7、in ati on of acce ptable limits for such imp urities in active substa nces is a difficult issue and not addressed in sufficie nt detail in the exist ing ICH Q3X guida nces. The data set usually available for geno toxic imp urities is quite variable and is the main factor that dictates the p rocess u

8、sed for the assessme nt of acce ptable limits. In the abse nee of data usually n eeded for the app licati on of one of the established risk assessme nt methods, i.e. data from carci nogeni city Ion g-term studies or data pro vid ing evide nee for a threshold mecha nism of geno toxicity, i mp leme nt

9、ati on of a gen erally app licable app roach as defi ned by the Threshold of Toxicological Concern (TTC) is propo sed. A TTC value of 1.5 in take of a geno toxic imp urity is con sidered to be associated with an acce ptable risk (excess cancer risk of <1 in 100,000 over a lifetime) for most p har

10、maceuticals. From this threshold value, ap ermitted level in the active substa nee can be calculated based on the exp ected daily dose. Higher limits may be justified un der certa in con diti ons such as short-term exp osure p eriods.基因毒性杂质的毒理学评估和这些杂质在活性药物中的可接受标准的测定是一件困难的 事情，并且在现有的ICH Q3X指南中也没有详细的规定

11、。现有的关于基因毒性杂质的相关 数据是容易变化的，也是对杂质可接受标准如何进行评价的主要影响因素。如果缺少风险评 估方法所需要的数据，比如，致癌作用的长期研究数据，或为基因毒性的阀值提供证据的数 据，一般建议使用一般通用的被定义为毒理学关注的阈值（TTC）的方法。一个“ 1$g/day” 的TTC值，即相当于每天摄入1.5卩g的基因毒性杂质，被认为对于大多数药品来说是可以 接受的风险（一生中致癌的风险小于十万分之 1）。按照这个阀值，可以根据这个预期的每日 摄入量计算出活性药物中可接受的杂质水平。较高的临界值可以在特定的条件下，如短期暴 露周期等，进行推算。1. INTRODUCTION 介绍

12、A gen eral concept of qualificati on of imp urities is described in the guideli nes for active substa nces (Q3A, I mp urities in New Active Substa nces) or medic inal p roducts (Q3B, I mp urities in New Medic inal P roducts), whereby qualificati on is defi ned as the p rocess of acquiri ng and evalu

13、at ing data that establishes the biological safety of an in dividual imp urity or a give n imp urity p rofile at the level(s) sp ecified. In the case of imp urities with a geno toxic poten tial, determ in ati on of acce ptable dose levels is gen erally con sidered as a p articularly critical issue,

14、which is not sp ecifically covered by the exist ing guideli nes.在原料药（Q3A）和药物制剂（Q3B）的杂质指导原则中，杂质限度确定的依据包括各 个杂质的生物安全性数据或杂质在某特定含量水平的研究概况。而对于遗传毒性杂质限度的确定，通常都认为是特别关键的问题，但目前尚无相关的指导原则。2. SCOPE 范围This Guideli ne describes a gen eral framework and p ractical app roaches on how to deal with geno toxic imp uriti

15、es in new active substa nces. It also relates to new app licati ons for existi ng active substa nces, where assessme nt of the route of syn thesis, p rocess con trol and impu rity p rofile does not pro vide reas on able assura nee that no new or higher levels of geno toxic imp urities are in troduce

16、d as comp ared to p roducts curre ntly authorised in the EU containing the same active substa nee. The same also app lies to variatio ns to exist ing Market ing Authorisati ons p erta ining to the syn thesis. The guideli ne does, however, not n eed to be app lied retros pectively to authorisedp rodu

17、cts uni ess there is a sp ecific cause for concern.本指导原则阐述了如何处理新原料药中遗传毒性杂质的一般框架和实际方法。该指导原 则也适用于已有原料药的新申请，如果其合成路线、过程控制和杂质研究尚无法确保不会产 生新的或更高含量的遗传毒性杂质（与 EU目前批准的相同原料药相比）。该指导原则同样 适用于已上市原料药有关合成方面的补充申请。除非有特殊原因，本指导原则不适用于已上 市的产品。In the curre nt con text the classificati on of a compound (impu rity) as geno to

18、xic in gen eral means that there are p ositive findings in established in vitro or in vivo geno toxicity tests with the main focus on DNA reactive substa nces that have a poten tial for direct DNA damage. Isolated in vitro findings may be assessed for in vivo releva nee in adequate follow- up test i

19、ng. In the abse nee of such in formatio n in vitro geno toxica nts are usually con sidered as p resu mp tive in vivo mutage ns and carci nogens.目前对于基因毒性杂质的分类主要是指：在以 DNA反应物质为主要研究对象的体内体 外试验中，如果发现它们对 DNA有潜在的破坏性，那可称之为基因毒性。如果有足够的后 续试验，可由单独的体外试验结果，对它的体内关联性进行评估。在缺乏这样的信息时，体 外基因毒性物质经常被考虑为假定的体内诱变剂和致癌剂。3. LEGA

20、L BASIS 法规依据This guideli ne has to be read in conjun cti on with Directive 2001/83/EC (as ame nded) and all releva nt CHMP Guida nee docume nts with sp ecial emp hasis on:在阅读该指南时有必要参考 “Directive 2001/83/EC以及相关的CHMP指南文件，特别是 以下几个指南：Imp urities Testi ng Guideli ne: Imp urities in New Drug Substa nces (

21、CPMP/ICH/2737/99,ICHQ3A(R)Note for Guida nee on Imp urities in New Drug P roducts (CPMP/ICH/2738/99, ICHQ3B (R)Note for Guida nee on Imp urities: Residual Solve nts (CPMP/ICH/283/95)Note for Guida nee on Geno toxicity: Guida nee on Sp ecific Asp ects of Regulatory Geno toxicityTests for P harmaceuti

22、cals (CPMP/ICH/141/95, ICHS2A)Note for Guida nee on Geno toxicity: A Sta ndard Battery for Geno toxicity Testi ng ofP harmaceuticals (CPMP/ICH/174/95, ICHS2B)4. TOXICOLOGICAL BACKGROUND毒理学背景Accordi ng to curre nt regulatory p ractice it is assumed that (in vivo) geno toxic compounds have the poten t

23、ial to damage DNA at any level of exp osure and that such damage maylead/c on tribute to tumour devel opment. Thus for geno toxic carc inogens it is p rude nt to assume that there is no discer nible threshold and that any level of exp osure carries a risk.根据目前的研究实践，具有（体内）遗传毒性的化合物在任何暴露量下都有可能对 DNA 产生损

24、伤，而这种损伤可能会引发肿瘤。因此，对于遗传毒性致癌物质，应谨慎认为不存在 明确的阈值，任何暴露量下都存在风险。However, the existe nee of mecha ni sms lead ing to biologically mea nin gful threshold effects is in creas in gly ack no wledged also for geno toxic eve nts. This holds true in p articular for compounds in teract ing with non-DNA targets and a

25、lso for poten tial mutage ns, which are rap idly detoxified before coming into con tact with critical targets. The regulatory app roach to such chemicals can be based on the ide ntificatio n of a critical no-observed-effect level (NOEL) and use of un certa inty factors.然而，对于一些遗传毒性事件，其产生生物学意义的阈值效应的机理

26、正越来越为人所了 解。对于非DNA靶点的化合物和潜在致突变剂更是如此，因为它们在与关键靶点接触前就 已经去毒化了。对于这些化合物，研究的基础可以是确定关键的未观察到影响的剂量 （NOEL） 和米用不确定因子。Eve n for compounds which are able to react with the DNA molecule, extra po lati on in a lin ear manner from effects in high-dose studies to very low level (huma n) expo sure may not be justified

27、due to several p rotective mecha ni sms op erat ing effectively at low doses. However, at p rese nt it is extremely difficult to exp erime ntally prove the existe nee of threshold for the geno toxicity of a give n mutage n. Thus, in the abse nee of approp riate evide nee supporting the existe nee of

28、 a threshold for a geno toxic compound making it difficult to defi ne a safe dose it is n ecessary to adopt a concept of a level of exp osure that carries an acce ptable risk.即使对能与DNA分子发生反应的化合物，由于低剂量时有多种有效的保护机制存在， 而不能将高剂量下的影响以线性方式外推到很低的（人）暴露水平。不过，目前要用实验方 法证明某诱变剂的遗传毒性阈值仍然非常困难。所以，在缺乏恰当的证据支持遗传毒性阈值 存在的情

29、况下，确定安全剂量很困难，因此非常有必要采用一个可接受风险的暴露水平概念。5. RECOMMENDATIONS 建议As stated in the Q3A guideli ne, actual and poten tial imp urities most likely to arise duri ng syn thesis, p urificati on and storage of the new drug substa nee should be ide ntified, based on a sound scie ntific app raisal of the chemical r

30、eact ions invo Ived in the syn thesis, i mp urities associated with raw materials that could con tribute to the impu rity p rofile of the new drug substa nee, and p ossible degradati on p roducts. This discussi on can be limited to those imp urities that might reas on ably be exp ected based on kno

31、wledge of the chemical reacti ons and con diti ons invo Ived. Guided by existi ng geno toxicity data or the p rese nee of structural alerts, poten tial geno toxic imp urities should be ide ntified. Whe n a poten tial imp urity contains structural alerts, additi onal geno toxicity test ing of the imp

32、 urity, typ ically in a bacterial reverse mutatio n assay, should be considered (Dobo et al. 2006, M ller et ali2006). While according to the Q3A guideline such studies can usually be con ducted on the drug substa nee containing the impu rity to be con trolled, studies using isolated imp urities are

33、 much more approp riate for this purpose and highly recomme nded.正如Q3A指导原则所述，根据合理的化学反应机理分析，在新的原料药合成、纯化和贮 存过程中很有可能产生实际的和潜在的杂质。依据现有的可能引起遗传毒性的结构”数据库，潜在的遗传毒性杂质应能被确认。如果潜在的杂质含有可引起遗传毒性的结构单元，该 杂质应考虑进行遗传毒性试验（一般是细菌回复突变试验）（Dobo等，2006）。虽然Q3A指导原则认为这些研究采用含有那些需控制杂质的原料药进行是可行的，但用分离出来的杂质进行这些研究更恰当，也是高度推荐的方法。For determ

34、 in ati on of acce ptable levels of expo sure to geno toxic care inogens con siderati ons of p ossible mecha ni sms of acti on and of the dose-res ponse relati onship are imp orta nt componen ts.Based on the above con siderati ons geno toxic imp urities may be dist in guished into the follow ing two

35、 classes:根据以上论述，遗传毒性杂质可以归纳成以下两类:-Geno toxic compounds with sufficie nt (ex perime ntal) evide nee for a threshold-related mecha nism有充分阈值相关机理证据（实验）的遗传毒性化合物-Geno toxic compounds without sufficie nt (ex perime ntal) evide nee for a threshold-related mecha nism无充分阈值相关机理证据（实验）的遗传毒性化合物5.1 Geno toxic Comp

36、ounds With Sufficie nt Evide nee for a Threshold-Related Mecha nism具有充分证据证明其阈值相关机理的基因毒性化合物Exa mples of mecha ni sms of geno toxicity that may be dem on strated to lead to non-I in ear or thresholded dose-res ponse relati onships in clude in teracti on with the spin dle app aratus of cell divisi on l

37、ead ing to aneupi oidy, topo isomerase in hibiti on, in hibiti on of DNA syn thesis, overloadi ng of defe nee mecha ni sms, metabolic overload and p hysiological p erturbati ons (e.g. i nductio n of erythro po eisis, hyper- or hyp othermia).非线性或阈值明确的剂量效应关系的遗传毒性机理包括：与细胞分化过程中纺锤体相互 作用；拓扑异构酶抑制；DNA合成抑制；过

38、度的防御机制；代谢过度和生理性干扰（如诱 导红血球生成，高体温和低体温）。For (classes of) compounds with clear evide nee for a thresholded geno toxicity, exp osure levels which are without app reciable risk of geno toxicity can be established accord ing to the p rocedure as outli ned for class 2 solve nts in the Q3C Note for Guida nee

39、 on Imp urities: Residual Solve nts. This approach calculates a“ Permitted Daily Exposure ” (PDE), which is derived from the NOEL, or tlowestobserved effect level (LOEL) in the most releva nt (ani mal) study using (UF).“un certa inty factors有明确遗传毒性阈值的化合物，不产生遗传毒性风险的暴露水平可以被确定，方法可参 照Q3C“杂质指导原则”中二类溶剂的限度

40、确定方法。该方法可计算每日最大允许暴露量”（PDE），数据来源于 不确定因数”动物研究中的NOEL （未观察到效果的最低水平）或观 察到效果的最低水平（LOEL ）。5.2 Geno toxic Compounds Without Sufficie nt Evide nee for a Threshold-RelatedMechanism不具备充分证据支持其阈值相关机理的基因毒性化合物aasThe assessme nt of acce ptability of geno toxic imp urities for which no threshold mecha ni sms are ide

41、 ntified should in clude both p harmaceutical and toxicological evaluati ons. In gen eral, p harmaceuical measureme nts should be guided by a p olicy of con trolli ng levels to reas on ably p racticable” (ALAR P principl e), where avoid ing is not p ossible. Levels con sidered being con siste nt wit

42、h the ALARP principle follow ing p harmaceutical assessme nt should be assessed for acce ptability from a toxicological point of view (see decisi on tree & followi ng sectio ns).对于此类遗传毒性杂质，研究应包括药学和毒理学评估。总之，如果杂质无法避免，药学方面的控制应遵循 合理可行的最低限量”原则（ALARP原则）。符合ALARP原则的杂质 水平再经毒理学方面的进一步评估，以验证其合理性（见决策树和以下章节）。

43、521 P harmaceutical Assessme nt 药学评价Q3A( R)。A sp ecific discussi on -as part of the overall discussi on on imp urities (see Q3A(R)-should be pro vided in the app licati on with regard to imp urities with poten tial geno toxicity.申请材料应提供关于潜在遗传毒性杂质的特别讨论资料（见A rati on ale of the prop osed formulati on/m

44、anu facturi ng strategy should be pro vided based on available formulati on op ti ons and tech no logies. The app lica nt should highlight, withi n the chemical p rocess and imp urity p rofile of active substa nee, all chemical substa nces, used as reage nts or p rese nt as in termediates, or side-p

45、 roducts, known as geno toxic an d/or carc inogenic (e.g. alkylat ing age nts).” in terms需要根据现在的配方选择和技术， 提供证明所选的配方/生产策略合理性的证据。申请人 应在合成工艺和杂质研究部分重点指出所有的化学物质， 包括用到的试剂、中间体、副产物, 哪些是已知遗传毒性和/或致癌性物质（如烷化剂）。More gen erally, react ing substa nces ahsubsta nces which show“ alerti ng structure of geno toxicity w

46、hich are not shared with the active substa nee should be con sidered (see e.g. Dobo et al. 2006). Poten tial alter natives which do no t lead to geno toxic residues in the final p roduct, should be used if available.值得关注的是，虽然有些含有可能引起遗传毒性的结构” （alerting structure）的反应试 剂与最终活性物质并没有共同结构，但也要考虑它们的遗传毒性（see

47、e.g. Dobo et al. 2006。.如果有可能，应该对它们进行一些替代研究，以使最终产品中不会引入基因毒性残留。A justificati on n eeds to be p rovided that no viable alter native exists, i ncludi ng alter native routes of syn thesis or formulati ons, differe nt start ing materials. This might for in sta nee in clude cases where the structure, which

48、 is res pon sible for the geno toxic an d/or carci nogenic poten tial is equivale nt to that n eeded in chemical syn thesis (e.g. alkylati on react ion s).需要提供充分的论证来说明没有可行的替代方法存在，包括可替代的合成路线或配方， 不同的起始物料等。比如，应证明具有遗传毒性和/或致癌性的结构在化学合成中（如烷化反 应）是必需的。If a geno toxic imp urity is con sidered to be un avoidab

49、le in a drug substa nee, tech ni cal efforts (e.g. p urificati on ste ps) should be un dertake n to reduce the content of the geno toxic residues in the final p roduct in comp lia nee with safety n eeds or to a level as low as reas on ably p racticable (see safety assessme nt). Data on chemical stab

50、ility of reactive in termediates, reacta nts, and other components should be in cluded in this assessme nt.如果遗传毒性杂质在原料中不可避免，则应该采取适当的技术（如纯化步骤）降低该杂 质的含量，以满足安全性要求，或符合 合理可行的最低限量”原则（见安全评估）。药学评 估还应包括反应中间体、反应物和其它组件等的化学稳定性研究。Detectio n an d/or qua ntificati on of these residues should be done by state-of-th

51、e-art an alytical tech niq ues.应该使用比较先进的分析检测技术来检测和量化这些残留的杂质。522 Toxicological Assessme nt 毒理学评价The imp ossibility of defi ning a safe exp osure level (zero risk concept) for geno toxiccarci nogens without a threshold and the realizatio n that comp lete elim in ati on of geno toxic imp urities from d

52、rug substa nces is ofte n un achievable, requires imp leme ntati on of a concept of an acce ptable risk level, i.e. an estimate of daily huma n exp osure at and below which there is a n egligible risk to huma n health.鉴于在没有明确阈值的前提下定义安全暴露水平（零风险）是不可能的，且从原料药中 完全除去遗传毒性杂质经常是很难做到的，所以有必要提出一个可接受风险水平”（accept

53、able risk leve）的概念，比如估算一个 每日最大暴露量”值，低于该暴露量时就可以 忽略其对人体健康的风险。P rocedures for the derivatio n of acce ptable risk levels are con sidered in the Appen dix 3 of theQ3C Note for Guida nee on Imp urities: Residual Solve nts for Class 1 solve nts. However, these app roaches require availability of adequate

54、data from Ion g-term carci nogeni city studies.对于可接受风险水平的推导过程请参见 Q3C （杂质指南注释：一类溶液残留）中的附件 三。然而，应用这些方法必须有足够多的长期致癌性研究数据。In most cases of toxicological assessme nt of geno toxic imp urities on ly limited data from in vitro studies with the imp urity (e.g. Ames test, chromosomal aberratio n test) are ava

55、ilable and thus established app roaches to determ ine acce ptable in take levels cannot be app lied. Calculati on of“ safety mult ip les” vitimi data (e.g. Ames test) are con sidered inapprop riate for justificati on of acce ptable limits. Moreover, n egative carc inogeni city and geno toxicity data

56、 with the drug substa nee containing the impu rity at low ppm levels do not pro vide sufficie nt assura nee for sett ing acce ptable limits for the imp urity due to the lack of sen sitivity of this test ing app roach. Eve n potent mutage ns and carc inogens are most likely to remai n un detected whe

57、 n tested as part of the drug substa nee, i.e. at very low exp osure levels. A p ragmatic app roach is therefore n eeded which recog ni ses that the p rese nee of very low levels of geno toxic imp urities is not associated with an un acce ptable risk.大多数情况下，遗传毒性杂质的毒理学评估只是局限于杂质的体外研究(如Ames试验,染色体畸变试验)，

58、但这些方法并不适用于确定杂质可接受的摄入水平。也就是说，根据体 外数据(如Ames试验)计算杂质的 安全倍数(safety mult ip les)”进而确定可接受的限度， 是不合适的。此外，用含有较低(ppm级)杂质水平的原料药研究其致癌性和遗传毒性，即 使得出阴性结果也不足以确保该杂质限度的合理性，因为这种试验方法缺少必要的灵敏度。 有些具有很强致突变性和致癌性物质与原料药一起进行试验时，因为在非常低的暴露水平情况下，很有可能因为低于检测限而无法检出。所以，如果认识到含量非常低的遗传毒性杂质 不存在不可接受的风险”(unacceptable risi)，那么可以采取实用的方法来控制该杂质。523 AppI icatio n of a Threshold of Toxicological Co ncern毒理学相关的阈值应用A threshold of toxicological concern (TTC) has bee n devel oped to defi ne a com mon exp osure level for any un studied chemical that will not p ose a risk of sig nific