EMEA基因毒性雜質(zhì)限度指引

1、20060628 EMEA/CHMP/QWP/251344/2006基因毒性雜質(zhì)限度指南(中英文對照)London, 28 June 2006CPMP/SWP/5199/02EMEA/CHMP/QWP/251344/2006The European Agency for the Evaluation of Medicinal Products歐洲共同體藥物評審委員會(EMEA)COMMITTEE FOR MEDICINAL PRODUCTS FOR HUMAN USE人用約品委員會(CHMP)GUIDLINE ON THE LIMITS OF GENOTOXIC IMPURITIES基因毒性雜

2、質(zhì)限度指南DESCUSSION IN THE SAFETY WORKING P/平安工作組之內(nèi)的討論XRTJJune 2002-October 2002TRANSMISSION TO CPMPCPM赧遞December 2002RELEASE FOR CONSULTATION專家討論December 2002DEADLINE FOR COMMENTS建議收集最后期限March 2003DISCUSSIONN THESAFETYVORKINPARTYKNtQUALITY WORKING PARTY平安工作組和質(zhì)量工作組之間的討論D June 2003-February 2004TRANSMISS

3、ION TO CPMPMarch 2004轉(zhuǎn)移給CPMPRE-RELEASE FOR CONSULTATIONJune 2004再次放行給參謀團(tuán)DEADLINE FOR COMMENTSDecember 2004收集意見的最后期限D(zhuǎn)ISCUSSION THESAFETYWORKINPARTAND February 2005-May 2006QUALITY WORKING PARTY平安工作組和質(zhì)量工作組之間的討論ADOPTION BY CHMP28 June 2006被CHM睬用DATE FOR COMING INTO EFFECT01January 2007生效日期KEYWORDS Impu

4、rities; Genotoxicity; Threshold of toxicological關(guān)鍵詞concern (TTC); Structure activity relationship (SAR)GUIDLINE ON THE LIMITS OF GENOTOXIC IMPURITIES基因毒性雜質(zhì)限度指南TABLE OF CONTENTS 錄EXECUTIVE SUMMARY 摘要31. INTRODUCTION紹32. SCOPE 范圍33. LEGAL BASIS 法律依 據(jù)34. TOXICOLOGICAL BACKGROUND 學(xué)背景45. RECOMMENDATION議4

5、5.1 Genotoxic CompoundSWith Sufficient Evidence for a Threshold-Related Mechanism具有充分證據(jù)證實其閾值相關(guān)機(jī)理的基因毒性化合物45.2 Genotoxic Compounds Without Sufficient Evidence for a Threshold-Related Mechanism不具備充分證據(jù)支持其閾值相關(guān)機(jī)理的基因毒性化合物55.2.1 Pharmaceutical Assessment 藥學(xué)評 價.55.2.2 Toxicological Assessment毒理學(xué)評價.55.2.3 App

6、lication of a Threshold of Toxicological Concern毒理學(xué)擔(dān)憂閾值應(yīng)用55.3 Decision Tree for Assessment of Acceptability of Genotoxic Impurities基因毒性雜質(zhì)可接受性評價決策樹7REFERENCE爹考文獻(xiàn)8EXECUTIVE SUMMARY 摘要The toxicological assessment of genotoxic impurities and the determination of acceptable limits for such impurities in

7、active substances is a difficult issue and not addressed in sufficient detail in the existing ICH Q3X guidances. The data set usually available for genotoxic impurities is quite variable and is the main factor that dictates the process used for the assessment of acceptable limits. In the absence of

8、data usually needed for the application of one of the established risk assessment methods, i.e. data from carcinogenicity long-term studies or data providing evidence for a threshold mechanismof genotoxicity, implementation of a generally applicable approach as defined by the Threshold of Toxicologi

9、cal Concern (TTC) is proposed.A TTC value of 1.5 仙 g/day intake of a genotoxic impurity is considered to be associated with an acceptable risk (excess cancer risk of <1 in 100,000 over a lifetime) for most pharmaceuticals. From this threshold value, a permitted level in the active substance can b

10、e calculated based on the expected daily dose. Higher limits maybe justified under certain conditions such as short-term exposure periods.基因毒性雜質(zhì)的毒理學(xué)評估和這些雜質(zhì)在活性藥物中的可接受標(biāo)準(zhǔn)的測定是一件困難的 事情,并且在現(xiàn)有的ICH Q3X指南中也沒有詳細(xì)的規(guī)定.現(xiàn)有的關(guān)于基因毒性雜質(zhì)的相關(guān)數(shù) 據(jù)是容易變化的,也是對雜質(zhì)可接受標(biāo)準(zhǔn)如何進(jìn)行評價的主要影響因素.如果缺少風(fēng)險評估 方法所需要的數(shù)據(jù),比方,致癌作用的長期研究數(shù)據(jù),或為基因毒性的閥值提供證據(jù)的

11、數(shù)據(jù), 一般建議使用一般通用的被定義為毒理學(xué)關(guān)注的閾值(TTQ的方法.一個“ 1.5仙g/day的TTC值,即相當(dāng)于每天攝入1.5 pg的基因毒性雜質(zhì),被認(rèn)為對于大多數(shù)藥品來說是可以接 受的風(fēng)險(一生中致癌的風(fēng)險小于十萬分之1).根據(jù)這個閥值,可以根據(jù)這個預(yù)期的每日攝 入量計算出活性藥物中可接受的雜質(zhì)水平.較高的臨界值可以在特定的條件下,如短期暴露 周期等,進(jìn)行推算.1. INTRODUCTION 介紹A general concept of qualification of impurities is described in the guidelines for active substa

12、nces (Q3A, Impurities in New Active Substances) or medicinal products (Q3B, Impurities in New Medicinal Products), whereby qualification is defined as the process of acquiring and evaluating data that establishes the biological safety of an individual impurity or a given impurity profile at the leve

13、l(s) specified. In the case of impurities with a genotoxic potential, determination of acceptable dose levels is generally considered as a particularly critical issue, which is not specifically covered by the existing guidelines.在原料藥(Q3A和藥物制劑(Q3B的雜質(zhì)指導(dǎo)原那么中,雜質(zhì)限度確定的依據(jù)包括各個 雜質(zhì)的生物平安性數(shù)據(jù)或雜質(zhì)在某特定含量水平的研究概況.而對于

14、遺傳毒性雜質(zhì)限度確實定,通常都認(rèn)為是特別關(guān)鍵的問題,但目前尚無相關(guān)的指導(dǎo)原那么.2. SCOPE 范圍This Guideline describes a general framework and practical approaches on how to deal with genotoxic impurities in new active substances. It also relates to new applications for existing active substances, where assessment of the route of synthesis,

15、process control and impurity profile does not provide reasonable assurance that no new or higher levels of genotoxic impurities are introduced as compared to products currently authorised in the EU containing the same active substance. The samealso applies to variations to existing Marketing Authori

16、sations pertaining to the synthesis. The guideline does, however, not need to be applied retrospectively to authorised products unless there is a specific cause for concern.本指導(dǎo)原那么闡述了如何處理新原料藥中遺傳毒性雜質(zhì)的一般框架和實際方法.該指導(dǎo)原 那么也適用于已有原料藥的新申請,如果其合成路線、過程限制和雜質(zhì)研究尚無法保證不會產(chǎn) 生新的或更高含量的遺傳毒性雜質(zhì)(與EU目前批準(zhǔn)的相同原料藥相比).該指導(dǎo)原那么同樣適用于已

17、上市原料藥有關(guān)合成方面的補(bǔ)充申請.除非有特殊原因,本指導(dǎo)原那么不適用于已上市 的產(chǎn)品.In the current context the classification of a compound(impurity) as genotoxic in general meansthat there are positive findings in established in vitro or in vivo genotoxicity tests with the main focus on DNA reactive substances that have a potential for di

18、rect DNA damage. Isolated in vitro findings may be assessed for in vivo relevance in adequate follow-up testing. In the absence of such information in vitro genotoxicants are usually considered as presumptive in vivo mutagens and carcinogens.目前對于基因毒性雜質(zhì)的分類主要是指：在以DN版應(yīng)物質(zhì)為主要研究對象的體內(nèi)體外試驗中,如果發(fā)現(xiàn)它們對DNAt潛在的破壞

19、性,那可稱之為基因毒性.如果有足夠的后續(xù)試 驗,可由單獨的體外試驗結(jié)果,對它的體內(nèi)關(guān)聯(lián)性進(jìn)行評估.在缺乏這樣的信息時,體外基 因毒性物質(zhì)經(jīng)常被考慮為假定的體內(nèi)誘變劑和致癌劑.3. LEGAL BASIS法規(guī)依據(jù)This guideline has to be read in conjunction with Directive 2001/83/EC (as amended) and all relevant CHMP Guidance documents with special emphasis on:在閱讀該指南時有必要參考“ Directive 2001/83/EC以及相關(guān)的CHMP旨南

20、文件,特別 是以下幾個指南：Impurities Testing Guideline: Impurities in New Drug Substances (CPMP/ICH/2737/99, ICHQ3A(R)Note for Guidance on Impurities in New Drug Products (CPMP/ICH/2738/99, ICHQ3B (R)Note for Guidance on Impurities: Residual Solvents (CPMP/ICH/283/95)Note for Guidance on Genotoxicity: Guidance

21、on Specific Aspects of Regulatory Genotoxicity Tests for Pharmaceuticals (CPMP/ICH/141/95, ICHS2A)Note for Guidance on Genotoxicity: A Standard Battery for Genotoxicity Testing of Pharmaceuticals (CPMP/ICH/174/95, ICHS2B)4. TOXICOLOGICAL BACKGROUND里學(xué)背景According to current regulatory practice it is a

22、ssumedthat (in vivo) genotoxic compounds have the potential to damage DNA at any level of exposure and that such damage may lead/contribute to tumour development. Thus for genotoxic carcinogens it is prudent to assume that there is no discernible threshold and that any level of exposure carries a ri

23、sk.根據(jù)目前的研究實踐,具有(體內(nèi))遺傳毒性的化合物在任何暴露量下都有可能對DNA產(chǎn)生損傷,而這種損傷可能會引發(fā)月中瘤.因此,對于遺傳毒性致癌物質(zhì),應(yīng)謹(jǐn)慎認(rèn)為不存在 明確的閾值,任何暴露量下都存在風(fēng)險.However, the existence of mechanisms leading to biologically meaningful threshold effects is increasingly acknowledged also for genotoxic events. This holds true in particular for compounds interact

24、ing with non-DNA targets and also for potential mutagens, which are rapidly detoxified before coming into contact with critical targets. The regulatory approach to such chemicals can be based on the identification of a critical no-observed-effect level (NOEL) and use of uncertainty factors.然而,對于一些遺傳

25、毒性事件,其產(chǎn)生生物學(xué)意義的閾值效應(yīng)的機(jī)理正越來越為人所了 解.對于非DNA巴點的化合物和潛在致突變劑更是如此,由于它們在與關(guān)鍵靶點接觸前就已 經(jīng)去毒化了.對于這些化合物,研究的根底可以是確定關(guān)鍵的未觀察到影響的劑量( NOEL 和米用不確定因子.Even for compounds which are able to react with the DNA molecule, extrapolation in a linear manner from effects in high-dose studies to very low level (human) exposure may not

26、be justified due to several protective mechanisms operating effectively at low doses. However, at present it is extremely difficult to experimentally prove the existence of threshold for the genotoxicity of a given mutagen. Thus, in the absence of appropriate evidence supporting the existence of a t

27、hreshold for a genotoxic compound making it difficult to define a safe dose it is necessary to adopt a concept of a level of exposure that carries an acceptable risk.即使對能與DN協(xié)子發(fā)生反響的化合物,由于低劑量時有多種有效的保護(hù)機(jī)制存在,而 不能將高劑量下的影響以線性方式外推到很低的(人)暴露水平.不過,目前要用實驗方法 證實某誘變劑的遺傳毒性閾值仍然非常困難.所以,在缺乏恰當(dāng)?shù)淖C據(jù)支持遺傳毒性閾值存 在的情況下,確定平安劑量很

28、困難,因此非常有必要采用一個可接受風(fēng)險的暴露水平概念.5. RECOMMENDATIONS®.As stated in the Q3A guideline, actual and potential impurities most likely to arise during synthesis, purification and storage of the new drug substance should be identified, based on a sound scientific appraisal of the chemical reactions involved

29、 in the synthesis, impurities associated with raw materials that could contribute to the impurity profile of the new drug substance, and possible degradation products. This discussion can be limited to those impurities that might reasonably be expected based on knowledge of the chemical reactions an

30、d conditions involved. Guided by existing genotoxicity data or the presence of structural alerts, potential genotoxic impurities should be identified. When a potential impurity contains structural alerts, additional genotoxicity testing of the impurity, typically in a bacterial reverse mutation assa

31、y, should be considered Dobo et al.2006, M u ller et al. 2006. While according to the Q3A guideline such studies canusually be conducted on the drug substance containing the impurity to be controlled, studies using isolated impurities are much more appropriate for this purpose and highly recommended

32、.正如Q3A旨導(dǎo)原那么所述,根據(jù)合理的化學(xué)反響機(jī)理分析,在新的原料藥合成、純化和貯 存過程中很有可能產(chǎn)生實際的和潛在的雜質(zhì).依據(jù)現(xiàn)有的“可能引起遺傳毒性的結(jié)構(gòu)數(shù)據(jù)庫,潛在的遺傳毒性雜質(zhì)應(yīng)能被確認(rèn).如果潛在的雜質(zhì)含有可引起遺傳毒性的結(jié)構(gòu)單元,該 雜質(zhì)應(yīng)考慮進(jìn)行遺傳毒性試驗一般是細(xì)菌回復(fù)突變試驗Dobo等,2006.雖然Q3A旨導(dǎo)原那么認(rèn)為這些研究采用含有那些需限制雜質(zhì)的原料藥進(jìn)行是可行的,但用別離出來的雜質(zhì)進(jìn)行這些研究更恰當(dāng),也是高度推薦的方法.For determination of acceptable levels of exposure to genotoxic carcinogens

33、considerations of possible mechanisms of action and of the dose-response relationship are important components. Based on the above considerations genotoxic impurities may be distinguished into the following two classes:根據(jù)以上論述,遺傳毒性雜質(zhì)可以歸納成以下兩類：-Genotoxic compounds with sufficient experimental evidence

34、 for a threshold-related mechanism有充分閾值相關(guān)機(jī)理證據(jù)實驗的遺傳毒性化合物-Genotoxic compounds without sufficient experimental evidence for a threshold-related mechanism無充分閾值相關(guān)機(jī)理證據(jù)實驗的遺傳毒性化合物5.1 Genotoxic Compounds With Sufficient Evidence for a Threshold-Related Mechanism具有充分證據(jù)證實其閾值相關(guān)機(jī)理的基因毒性化合物Examples of mechanisms o

35、f genotoxicity that may be demonstrated to lead to non-linear or thresholded dose-response relationships include interaction with the spindle apparatus of cell division leading to aneuploidy, topoisomerase inhibition, inhibition of DNAsynthesis, overloading of defence mechanisms, metabolic overload

36、and physiological perturbations (e.g. induction of erythropoeisis, hyper- or hypothermia).非線性或閾值明確的劑量效應(yīng)關(guān)系的遺傳毒性機(jī)理包括：與細(xì)胞分化過程中紡錘體相互 作用；拓?fù)洚悩?gòu)酶抑制；DN的成抑制；過度的防御機(jī)制；代謝過度和生理性干擾(如誘導(dǎo) 紅血球生成,高體溫和低體溫).For (classes of) compounds with clear evidence for a thresholded genotoxicity, exposure levels which are without ap

37、preciable risk of genotoxicity can be established according to the procedure as outlined for class 2 solvents in the Q3c Note for Guidance on Impurities: Residual Solvents. This approach calculates a" PermittedDaily Exposure (PDE), which is derived from the NOEL,or the lowestobserved effect lev

38、el (LOEL) in the most relevant (animal) study using "uncertainty factors " (UF).有明確遺傳毒性閾值的化合物,不產(chǎn)生遺傳毒性風(fēng)險的暴露水平可以被確定,方法可參 照Q3c雜質(zhì)指導(dǎo)原那么中二類溶劑的限度確定方法.該方法可計算“每日最大允許暴露量 (PDE,數(shù)據(jù)來源于“不確定因數(shù)動物研究中的 NOEL(未觀察到效果的最低水平)或觀察到效果的最低水平(LOEL.5.2 Genotoxic Compounds Without Sufficient Evidence for a Threshold-Relat

39、ed Mechanism不具備充分證據(jù)支持其閾值相關(guān)機(jī)理的基因毒性化合物The assessment of acceptability of genotoxic impurities for which no threshold mechanisms are identified should include both pharmaceutical and toxicological evaluations. In general, pharmaceutical measurements should be guided by a policy of controlling levels to

40、“as low as reasonably practicable (ALARP principle),where avoiding is not possible. Levels considered being consistent with the ALARP principle following pharmaceutical assessment should be assessed for acceptability from a toxicological point of view (see decision tree & following sections).對于此

41、類遺傳毒性雜質(zhì),研究應(yīng)包括藥學(xué)和毒理學(xué)評估.總之,如果雜質(zhì)無法防止,藥 學(xué)方面的限制應(yīng)遵循“合理可行的最低限量原那么(ALARPM那么).符合ALARPM那么的雜質(zhì)水平再經(jīng)毒理學(xué)方面的進(jìn)一步評估,以驗證其合理性(見決策樹和以下章節(jié)).5.2.1 Pharmaceutical Assessment 藥學(xué)評價A specific discussion as part of the overall discussion on impurities (see Q3A(R) should be provided in the application with regard to impurities w

42、ith potential genotoxicity.申請材料應(yīng)提供關(guān)于潛在遺傳毒性雜質(zhì)的特別討論資料(見Q3A(R)oA rationale of the proposed formulation/manufacturing strategy should be provided based on available formulation options and technologies. The applicant should highlight, within the chemical process and impurity profile of active substance,

43、 all chemical substances, used as reagents or present as intermediates, or side-products, known as genotoxic and/or carcinogenic (e.g. alkylating agents).需要根據(jù)現(xiàn)在的配方選擇和技術(shù),提供證實所選的配方/生產(chǎn)策略合理性的證據(jù).申請人 應(yīng)在合成工藝和雜質(zhì)研究局部重點指出所有的化學(xué)物質(zhì),包括用到的試劑、中間體、副產(chǎn)物,哪些是遺傳毒性和/或致癌性物質(zhì)(如烷化劑).More generally, reacting substances and sub

44、stan ces which show “alerting structure “ in terms of genotoxicity which are not shared with the active substance should be considered (see e.g. Dobo et al. 2006). Potential alternatives which do not lead to genotoxic residues in the final product, should be used if available.值得關(guān)注的是,雖然有些含有“可能引起遺傳毒性的

45、結(jié)構(gòu)(alerting structure )的反響試劑與最終活性物質(zhì)并沒有共同結(jié)構(gòu),但也要考慮它們的遺傳毒性(see e.g. Doboet al.2006). o如果有可能,應(yīng)該對它們進(jìn)行一些替代研究,以使最終產(chǎn)品中不會引入基因毒性殘 留.A justification needs to be provided that no viable alternative exists, including alternative routes of synthesis or formulations, different starting materials. This might for in

46、stance include cases where the structure, which is responsible for the genotoxic and/or carcinogenic potential is equivalent to that needed in chemical synthesis (e.g. alkylation reactions).需要提供充分的論證來說明沒有可行的替代方法存在,包括可替代的合成路線或配方, 不同的起始物料等.比方,應(yīng)證實具有遺傳毒性和/或致癌性的結(jié)構(gòu)在化學(xué)合成中(如烷化反響)是必需的.If a genotoxic impurity

47、 is considered to be unavoidable in a drug substance, technical efforts (e.g. purification steps) should be undertaken to reduce the content of the genotoxic residues in the final product in compliance with safetyneeds or to a level as low as reasonably practicable (see safety assessment). Data on c

48、hemical stability of reactive intermediates, reactants, and other components should be included in this assessment.如果遺傳毒性雜質(zhì)在原料中不可防止,那么應(yīng)該采取適當(dāng)?shù)募夹g(shù)(如純化步驟)降低該雜 質(zhì)的含量,以滿足平安性要求,或符合“合理可行的最低限量原那么(見平安評估).藥學(xué)評估還應(yīng)包括反響中間體、反響物和其它組件等的化學(xué)穩(wěn)定性研究.Detection and/or quantification of these residues should be done by state-o

49、f-the-art analytical techniques.應(yīng)該使用比擬先進(jìn)的分析檢測技術(shù)來檢測和量化這些殘留的雜質(zhì).5.2.2 Toxicological Assessment毒理學(xué)評價The impossibility of defining a safe exposure level (zero risk concept) for genotoxic carcinogens without a threshold and the realization that complete elimination of genotoxic impurities from drug substa

50、nces is often unachievable, requires implementation of a concept of an acceptable risk level, i.e. an estimate of daily humanexposure at and below which there is a negligible risk to humanhealth.鑒于在沒有明確閾值的前提下定義平安暴露水平(零風(fēng)險)是不可能的,且從原料藥中 完全除去遺傳毒性雜質(zhì)經(jīng)常是很難做到的,所以有必要提出一個“可接受風(fēng)險水平(acceptable risk level )的概念,比方

51、估算一個“每日最大暴露量值,低于該暴露量 時就可以忽略其對人體健康的風(fēng)險.Procedures for the derivation of acceptable risk levels are considered in the Appendix 3 of the Q3c Note for Guidance on Impurities: Residual Solvents for Class 1 solvents. However, these approaches require availability of adequate data from long-term carcinogeni

52、city studies.對于可接受風(fēng)險水平的推導(dǎo)過程請參見Q3c(雜質(zhì)指南注釋：一類溶液殘留)中的附件三.然而,應(yīng)用這些方法必須有足夠多的長期致癌性研究數(shù)據(jù).In most cases of toxicological assessment of genotoxic impurities only limited data from in vitro studies with the impurity (e.g. Amestest, chromosomal aberration test) are available and thus established approaches to de

53、termine acceptable intake levels cannot be applied. Calculation of“safety multiples “ from in vitro dat a(e.g. Amestest) are considered inappropriate for justification of acceptable limits.Moreover, negative carcinogenicity and genotoxicity data with the drug substance containing the impurity at low

54、 ppm levels do not provide sufficient assurance for setting acceptable limits for the impurity due to the lack of sensitivity of this testing approach. Even potent mutagens and carcinogens are most likely to remain undetected whentested as part of the drug substance, i.e. at very low exposure levels

55、.A pragmatic approach is therefore needed which recognises that the presence of very low levels of genotoxic impurities is not associated with an unacceptable risk.大多數(shù)情況下,遺傳毒性雜質(zhì)的毒理學(xué)評估只是局限于雜質(zhì)的體外研究如Ames試驗,染色體畸變試驗,但這些方法并不適用于確定雜質(zhì)可接受的攝入水平.也就是說,根據(jù)體 外數(shù)據(jù)如Ames試驗計算雜質(zhì)白“平安倍數(shù)safety multiples 、進(jìn)而確定可接受 的限度,是不適宜的.止

56、匕外,用含有較低ppm級雜質(zhì)水平的原料藥研究其致癌性和遺傳 毒性,即使得出陰性結(jié)果也缺乏以保證該雜質(zhì)限度的合理性,由于這種試驗方法缺少必要的 靈敏度.有些具有很強(qiáng)致突變性和致癌性物質(zhì)與原料藥一起進(jìn)行試驗時,由于在非常低的暴 露水平情況下,很有可能由于低于檢測限而無法檢出.所以,如果熟悉到含量非常低的遺傳 毒性雜質(zhì)不存在“不可接受的風(fēng)險 unacceptable risk ,那么可以采取實用的方法來控 制該雜質(zhì).5.2.3 Application of a Threshold of Toxicological Concern毒理學(xué)相關(guān)的閾值應(yīng)用A threshold of toxicologic

57、al concern (TTC) has been developed to define a common exposure level for any unstudied chemical that will not pose a risk of significant carcinogenicity or other toxic effects (Munro et al. 1999, Kroes and Kozianowski2002). This TTC value was estimated to be 1.5g g/person/day. The TTC, originallyde

58、veloped as a "threshold of regulation " at the FDA for food contact materials (Rulis 1989, FDA1995) was established based on the analysis of 343 carcinogens from a carcinogenic potency database (Gold et al. 1984) and was repeatedly confirmed by evaluations expanding the database to more than 700 carcinogens (Munro 1990, Cheeseman et al. 1999, Kroes et al. 2004). The probability distribution of carcinogenic potencies has been used to derive an estimate of a daily exposure level (n g/person) of most carcinogens which wo