吴殷囡;唐婉;尹菁;刘洋;汪玉馨;陆益红;

• 1:徐州医科大学药物分析教研室
• 2:江苏省食品药品监督检验研究院
• 3:南京鼓楼医院集团宿迁市人民医院
• 4:徐州医科大学宿迁附属医院

摘要(Abstract)：

目的评价萘普生的杂质acetylnerolin(Ace)的遗传毒性。方法采用基于定量构效关系的方法ADMET、Derek和Sarah,预测Ace的遗传毒性,利用体外染色体畸变和细菌反向突变(Ames)实验验证上述结果。在染色体畸变实验中,中国仓鼠肺(CHL)细胞与Ace 10,20,40 mg·L^(-1)在有/无代谢活化系统混合液(S9 mix)的条件下,分别以甲基磺酸甲酯(MMS)20 m L·L(-1)在有/无代谢活化系统混合液(S9 mix)的条件下,分别以甲基磺酸甲酯(MMS)20 m L·L^(-1)及环磷酰胺(CP)12 mg·L(-1)及环磷酰胺(CP)12 mg·L^(-1)作为阳性对照,孵育4 h后,对每个畸变中期(包括断裂、交换、环状、分裂和多倍体)的染色体数目计数并记录,当畸变率<5%为阴性,>10%为阳性。在Ames实验中,用5种鼠伤寒沙门氏菌(TA97,TA98,TA100,TA102和TA1535)对Ace进行致突变评价。这5种菌种分别与Ace 5,25,125和625μg(每个平板)在有无S9 mix条件下孵育48～72 h。在不添加S9 mix的条件下,敌克松50μg(每个平板)作为TA97和TA98组的阳性对照;MMS2.0μL(每个平板)作为TA100和TA102组的阳性对照;叠氮化钠1.5μg(每个平板)作为TA1535的阳性对照;在添加S9 mix的条件下,2-氨基芴100μg(每个平板)作为TA97,TA98和TA100的阳性对照;1,8-二羟基蒽醌100μg(每个平板)和CP 50μg(每个平板)分别作为TA102和TA1535的阳性对照,当菌落数≥2×阴性对照时,则认为该化合物具有致突变性,反之则为阴性。结果 Derek和Sarah软件预测NPX杂质不具有遗传毒性,但ADMET数据显示Ace可诱发染色体畸变。在染色体畸变实验中,Ace 40 mg·L(-1)作为阳性对照,孵育4 h后,对每个畸变中期(包括断裂、交换、环状、分裂和多倍体)的染色体数目计数并记录,当畸变率<5%为阴性,>10%为阳性。在Ames实验中,用5种鼠伤寒沙门氏菌(TA97,TA98,TA100,TA102和TA1535)对Ace进行致突变评价。这5种菌种分别与Ace 5,25,125和625μg(每个平板)在有无S9 mix条件下孵育48～72 h。在不添加S9 mix的条件下,敌克松50μg(每个平板)作为TA97和TA98组的阳性对照;MMS2.0μL(每个平板)作为TA100和TA102组的阳性对照;叠氮化钠1.5μg(每个平板)作为TA1535的阳性对照;在添加S9 mix的条件下,2-氨基芴100μg(每个平板)作为TA97,TA98和TA100的阳性对照;1,8-二羟基蒽醌100μg(每个平板)和CP 50μg(每个平板)分别作为TA102和TA1535的阳性对照,当菌落数≥2×阴性对照时,则认为该化合物具有致突变性,反之则为阴性。结果 Derek和Sarah软件预测NPX杂质不具有遗传毒性,但ADMET数据显示Ace可诱发染色体畸变。在染色体畸变实验中,Ace 40 mg·L^(-1)致畸变率>5%,但<10%;Ace <20 mg·L(-1)致畸变率>5%,但<10%;Ace <20 mg·L^(-1)时致畸变率<5%,提示Ace可能诱发染色体畸变,这与ADMET结果一致;Ames试验结果表明,与阴性对照组相比,5种菌种每个平板给药Ace 5～625μg后,各组的细菌数量没有显著增加,提示Ace无致突变性,这与ADMET结果相反。结论 Ace有潜在的致染色体畸变性,对于长期服用萘普生的患者,为保障用药安全,建议将Ace的含量从普通杂质的0.15%降至0.015%。

关键词(KeyWords)： 染色体畸变;定量构效关系;萘普生

Abstract:

Keywords:

基金项目(Foundation): 2017 National Evaluation Sampling Inspection Project;; Project of Chinese Pharmacopoeia Commission (2016-452)~~

作者(Authors): 吴殷囡;唐婉;尹菁;刘洋;汪玉馨;陆益红;

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