一步法分析细胞色素P450 2D6酶活性缺陷等位基因CYP2D6A和CYP2D6B

目的：ＣＹＰ２Ｄ６Ａ和ＣＹＰ２Ｄ６Ｂ是引起细胞色素Ｐ４５０２Ｄ６（ＣＹＰ２Ｄ６）酶活性缺陷的最主要的等位基因，对ＣＹＰ２Ｄ６Ａ和ＣＹＰ２Ｄ６Ｂ的检测可准确（＞９２％）预测ＣＹＰ２Ｄ６慢代谢者。本研究利用等位基因特异扩增法，建立了一步ＰＣＲ法测定ＣＹＰ２Ｄ６Ａ和ＣＹＰ２Ｄ６Ｂ等位基因。方法：等位基因特异扩增法分析ＣＹＰ２Ｄ６Ａ和ＣＹＰ２Ｄ６Ｂ等位基因；右美沙芬作为探针药物测定表型。结果：经１３０例测定，说明本法更为快捷、更少污染。结论：本法的建立为该项测定应用于临床、指导临床合理用药奠定基础     

364 78种生药提取物对人肝脏微粒体 CYP3A4和CYP2D6的抑制作用

探讨了78种生药提取物对人肝脏微粒体的药物代谢酶细胞色素P450（CYP）CYP3A4和CYP2D6的抑制作用。     

桂枝汤对大鼠肝微粒体细胞色素P450酶活性的影响

目的:研究桂枝汤对大鼠肝微粒体细胞色素P450酶CYP1A2、CYP2D6和CYP3A4活性的影响。方法:18只大鼠被随机分成3组,每组6只。以生理盐水为空白对照,大鼠每日灌胃给予桂枝汤10 g/kg,连续7 d,测定其肝微粒体CYP1A2、CYP2D6和CYP3A4活性。结果:与对照组相比,桂枝汤对大鼠CYP1A2、CYP2D6和CYP3A4的活性无明显差异(P>0.05)。结论:桂枝汤对大鼠CYP1A2、CYP2D6和CYP3A4活性无影响。     

26种中药提取物对CYP3A4和CYP2D6代谢的抑制作用

〔英〕/IwataH…∥JTradMed.-2004,21(6).-281～286研究认为,细胞色素P450(CYP)酶与药物相互作用。本次在NADPH-生成体系存在的情况下,以单味中药甲醇提取物对人肝细胞微粒体进行预培养,检测26种中药对CYP3A4和CYP2D6的抑制作用。材料和方法:①中药甲醇提取物的制备:26种中药(白芷、苍术、肉桂、丁香、黄连、延胡索、干姜、麻黄、吴茱萸、连翘、钩藤、牛蒡子、羌活、甘草、枇杷叶、厚朴、牡丹皮、黄柏、大黄、苏木、五味子、黄芩、防己、辛夷、前胡、山椒)的粉末状水提取物(1g)用甲醇提取,1μL甲醇可溶性组分相当于0.33mg中药水提…     

丹皮酚对大鼠肝微粒体细胞色素P450酶含量及活性的影响

目的:考察丹皮酚对大鼠肝微粒体的蛋白含量、CYP450酶总量和主要CYP450酶亚型(CYP1A2、CYP2D6、CYP3A4和CYP2C19)活性的影响。方法:大鼠灌胃给予丹皮酚(100 mg/(kg·d)-1,连续7 d,测定其肝微粒体蛋白含量、CYP450蛋白含量以及CYP1A2、CYP2D6、CYP3A4及CYP2C19活性。结果与空白对照组相比,丹皮酚给药组大鼠肝微粒体蛋白含量及肝微粒体CYP450含量无明显差异(P>0.05)。丹皮酚给药后,给药组大鼠的平均CYP2D6活性是对照组的2倍;而二两组之间CYP1A2、CYP3A4和CYP2C19的活性相当。结论丹皮酚对大鼠CYP2D6活性有一定诱导作用,对CYP1A2、CYP3A4和CYP2C19的活性没有影响。     

应用大鼠肝微粒体筛选对细胞色素P4502D6有抑制作用的中药

目的从17种中药(单体、提取物和注射液)中快速筛选出对细胞色素P4502D6(CYP2D6)亚型有抑制作用的中药,建立高通量筛选对CYP2D6有抑制作用中药的技术平台。方法取大鼠空白肝微粒体,分别加入17种中药,HPLC法测定大鼠肝微粒体中CYP2D6的探针药物右美沙芬(DM)的代谢率,通过代谢率降低的现象初步筛选出能抑制肝微粒体中CYP2D6代谢的药物。结果槲皮素、黄芩苷、厚朴提取物、清开灵注射液、盐酸川芎嗪注射液可明显降低DM的代谢率(P<0.01)。结论初步筛选出槲皮素、黄芩苷、厚朴提取物、清开灵注射液、盐酸川芎嗪注射液这5种中药可抑制肝微粒体CYP2D6酶活性。此方法可作为高通量筛选对CYP2D6活性有抑制作用中药的技术平台。     

中国汉族和蒙古族健康人药物代谢酶CYP3A4、CYP2C9、CYP2C19、CYP2D6基因多态性分析

 目的 用基因分型技术对中国汉族、蒙古族健康人CYP3A4、CYP2C9、CYP2C19、CYP2D6进行基因多态性分析,并对汉族人和蒙古族人基因表型和基因频率进行比较。方法 聚合酶链反应-限制性片段长度多态性(PCR-RFLP)对CYP3A4、CYP2C9、CYP2C19、CYP2D6进行分型。结果 汉族、蒙古族健康人CYP3A4*5等位基因频率为0, CYP3A4*18等位基因频率分别为0.183 8、0.202 5,CYP2C9*2等位基因频率分别为0.011 0、0.025 3, CYP2C9*13等位基因频率分别为0、0.00 32,CYP2C19*2等位基因频率分别为0.386 0、0.4146, CYP2C19*3等位基因频率分别为0.051 5、0.044 3,CYP2D6*10等位基因频率分别为0.573 5、0.465 2。结论 汉族、蒙古族健康人群CYP3A4*18、CYP2C19*2、CYP2C19*3、CYP2D6*10等位基因频率均没有显著性差异;本试验在汉族、蒙古族健康人中未发现CYP3A4*5等位基因;仅在蒙古族健康受试人群中发现1人为CYP2C9*1/*13基因型;蒙古族CYP2C9*2等位基因频率远小于汉族（P=0.023）。     

红花注射液对大鼠细胞色素P450 2D6亚型的抑制作用

目的研究红花注射液对大鼠细胞色素P450 2D6亚型(CYP2D6)的影响。方法利用探针药物右美沙芬(DM),高效液相色谱法测定各实验组(对照组,红花注射液0.9、1.8、3.6 mL/kg组)大鼠体内尿液中与体外肝微粒体温孵系统中DM的代谢率,考察红花注射液对大鼠CYP2 D6活性的影响。结果体内实验和体外实验中,红花注射液1.8和3.6 mL/kg组DM的代谢率均明显低于对照组(P<0.05、0.01);抑制实验中,体外肝微粒体温孵系统中红花注射液组和西咪替丁组DM的代谢率明显低于空白组(P<0.05);含红花生药量为30 mg/mL时红花注射液组DM的代谢率与西咪替丁(0.6 mg/mL)时DM的代谢率相近,抑制能力相当;红花注射液的IC50为10.64 mg/mL。结论红花注射液对大鼠CYP2D6有显著的抑制作用。     

土茯苓6种黄酮类成分在CYP2D6中的体外酶促动力学研究

目的 比较土茯苓中6种黄酮类成分在CYP2D6中的体外酶促动力学过程.方法 以表儿茶素、新落新妇苷、落新妇苷、异落新妇苷、黄杞苷、白藜芦醇6种黄酮类成分为底物,研究了不同孵育时间、不同CYP2D6酶浓度对土茯苓中6种黄酮类成分体外代谢的影响.结果 6种黄酮类成分在30 ～ 90 min内呈线性消除,且代谢消除率随着CY...     

长春花中对细胞色素P4502D6有抑制作用的化合物

细胞色素P450（CYP）酶的失活是药物相互作用的主要原因之一。作者研究了长春花Catharanthus roseeus中对CYP3A4和CYP2D6介导的代谢具抑制活性的成分，以及这些成分的作用机制。     

清开灵注射液对大鼠CYP1A2和2D6的影响

目的：通过清开灵注射液的大鼠体内、外实验,观察清开灵注射液对大鼠CYP1A2亚型,CYP2D6亚型的影响。方法：通过HPLC法测定全血中咖啡因的代谢率,观测清开灵注射液对大鼠CYP1A2活性的影响；通过HPLC法测定大鼠肝微粒体重组系统非那西丁的代谢比率,确定清开灵注射液对大鼠肝微粒体CYP1A2亚型的作用；测定大鼠肝微粒体重组系统右美沙芬的代谢比率,确定清开灵注射液对大鼠肝微粒体CYP2D6亚型的作用。结果：实验组中给予大鼠不同浓度的清开灵注射液(0.15,0.3,0.6 mL·kg^-1),其咖啡因代谢率为(15.9±3.8)%,(14.5±1.8)%,(12.3±1.2)%,对照组为(16.8±5.9)%,各剂量组及对照组间均无显著性差异；肝微粒体体外重组系统中,实验组各浓度清开灵注射液对CYP2D6没有影响；高剂量组清开灵注射液对CYP1A2有抑制作用。结论：清开灵注射液对CYP1A2和 CYP2D6的活性没有影响。     

Effects of Panax notoginseng saponins on the activities of CYP1A2, CYP2C9, CYP2D6 and CYP3A4 in rats in vivo

The aim of this study was to assess the influence of the Panax notoginseng saponins (PNS) on the activities of the drug‐metabolizing enzymes cytochrome P450 (CYP450) 1A2, 2 C9, 2D6 and 3A4 in rats. The activities of CYP1A2, 2 C9, 2D6 and 3A4 were measured using specific probe drugs. After pretreatment for 1 week with PNS or physiological saline (control group), probe drugs caffeine (10 mg/kg; CYP1A2 activity), tolbutamide (15 mg/kg; CYP2C9 activity), metoprolol (20 mg/kg; CYP2D6 activity) and dapsone (10 mg/kg; CYP3A4 activity) were administered to rats by intraperitoneal injection. The blood was then collected at different times for ultra performance liquid chromatography/tandem mass spectrometry (UPLC‐MS/MS) analysis. The data showed that PNS exhibited an induction effect on CYP1A2 by decreasing caffeine C_max (36.3%, p < 0.01) and AUC_0‐∞ (22.77%, p < 0.05) and increasing CL/F (27.03%, p < 0.05) compared with those of the control group. Western blot analysis was used to detect the effect of PNS on the protein level of CYP1A2, and the results showed that PNS could upregulate the protein expression of CYP1A2. However, no significant changes in CYP2C9, 2D6 or 3A4 activities were observed. In conclusion, the results indicate that PNS could induce CYP1A2, which may affect the disposition of medicines primarily dependent on the CYP1A2 pathway. Our work may be the basis of related herb–drug interactions in the clinic. Copyright © 2011 John Wiley & Sons, Ltd.     

Exploring the possible metabolism mediated interaction of Glycyrrhiza glabra extract with CYP3A4 and CYP2D6

The rhizome of Glycyrrhiza glabra L. (licorice) is used very widely in Indian and Chinese traditional medicine, and it is a popular flavor ingredient of drinks, sweets and candies. Its medicinal uses include treating bronchitis, dry cough, respiratory infections, liver disorders and diabetes. Glycyrrhizin is normally considered to be its biologically active marker, so a rapid RP‐HPLC method was developed for the quantitative estimation of glycyrrhizin in the extract. The effect of the standardized extract and its marker on drug metabolizing enzymes was evaluated through CYP3A4 and CYP2D6 inhibition assays to evaluate the safety through its drug interaction potential. The inhibition of CYP3A4 and CYP2D6 isozymes was analysed by the fluorescent product formation method. In the CYP450‐CO assay, the interaction potential of the standardized extract and pooled microsomes (percentage inhibition 23.23 ± 1.84%), was found to be less than the standard inhibitor. In the fluorimetric assay, G. glabra extracts showed higher IC₅₀ values than their positive inhibitors, ketoconazole and quinidine for CYP3A4 and CYP2D6, respectively. Furthermore, the interaction potential of the plant extract was greater than the pure compound. The results demonstrate that G. glabra and its principle bioactive compound, glycyrrhizin, when co‐administered with conventional medicines showed only a weak interaction potential with drug metabolizing enzymes. Copyright © 2011 John Wiley & Sons, Ltd.     

中国汉族、维吾尔族、哈萨克族人群中细胞因子的表达与CYP2D6基因多态性

 目的探讨细胞因子在汉族、维吾尔族、哈萨克族正常人血清中的表达水平是否存在民族间的差异以及与CYP2D6基因多态性是否有关。方法ELISA双抗体夹心法。结果血清中肿瘤坏死因子-α(TNF-α)和转化生长因子-β₁(TGF-β₁)浓度范围:汉族为18-665pg·L^-1和0.04-106ng·L^-1,维吾尔族为19～1020pg·L^-1和0.05～112ng·L^-1,哈萨克族为21～1014pg·L^-1和0.11～94ng·L^-1。结论TNF-α的浓度在汉族与维吾尔族间存在显著性差异(P<0.01),哈萨克族与维吾尔族间存在显著性差异(P<0.01),汉族与哈萨克族间无显著性差异(P>0.05)。TGF-β₁浓度在汉族与维吾尔族间存在显著性差异(P<0.01),汉族与哈萨克族间存在显著性差异(P<0.05),维吾尔族与哈萨克族之间无显著性差异。     

The in vitro inhibition of human CYP1A2, CYP2D6 and CYP3A4 by tetrahydropalmatine, neferine and berberine

The purpose of this study was to investigate the in vitro inhibition potential of the three purified herbal constituents tetrahydropalmatine (Tet), neferine (Nef) and berberine (Ber) towards recombinant human CYP1A2, CYP2D6 and CYP3A4 metabolic activities. In vitro incubations were performed with phenacetin, dextromethorphan and testosterone, respectively, as CYP substrates and their metabolites were determined by validated HPLC methods. Positive control inhibitors were run for each CYP in all incubation series. Inhibition was expressed by IC₅₀ values. All herbal constituents demonstrated some, but variable, inhibition potencies towards the investigated CYP enzymes. CYP2D6 was the most sensitive for inhibition and then mainly by Tet and Ber with IC₅₀ values of 3.04 ± 0.26 µm and 7.40 ± 0.36 µm , respectively. CYP3A4 and especially CYP1A2 were inhibited to a much smaller extent by all constituents. Neferine showed the lowest overall interaction potential towards the CYP enzymes investigated. The CYP inhibition potential for the purified constituents could be related to their chemical structures. No clinical significant metabolic interaction seems likely to occur between the CYP enzymes and herbal constituents tested, with a possible exception for the CYP2D6 inhibition by Tet and Ber. Copyright © 2011 John Wiley & Sons, Ltd.     

In vitro Inhibition of Human CYP1A2, CYP2D6, and CYP3A4 by Six Herbs Commonly Used in Pregnancy

Black elderberry, cranberry, fennel, ginger, horsetail, and raspberry leaf, herbs frequently used in pregnancy, were investigated for their in vitro CYP1A2, 2D6, and 3A4 inhibitory potential. Aqueous or ethanolic extracts were made from commercially available herbal products, and incubations were performed with recombinant cDNA‐expressed human CYP enzymes in the presence of positive inhibitory controls. Metabolite formation was determined by validated LCMS/MS or HPLC methodologies. IC₅₀ inhibition constants were estimated from CYP activity inhibition plots using non‐linear regression. The most potent inhibition was shown for fennel towards CYP2D6 and 3A4 with respective IC₅₀ constants of 23 ± 2 and 40 ± 4 µg/ml, horsetail towards CYP1A2 with an IC₅₀ constant of 27 ± 1 µg/ml, and raspberry leaf towards CYP1A2, 2D6, and 3A4 with IC₅₀ constants of 44 ± 2, 47 ± 8, and 81 ± 11 µg/ml, respectively. Based on the recommended dosing of the different commercial herbal products, clinically relevant systemic CYP inhibitions could be possible for fennel, horsetail, and raspberry leaf. In addition, fennel and raspberry leaf might cause a clinically relevant inhibition of intestinal CYP3A4. The in vivo inhibitory potential of these herbs towards specific CYP enzymes should be further investigated. Copyright © 2013 John Wiley & Sons, Ltd.     

Drug interaction potential of the seed extract of Urtica urens L. (dwarf Nettle)

Dwarf nettle (Urtica urens) seed extract was examined in vivo in the rat for its potential to modulate drug metabolizing enzymes including aminopyrine N‐demethylase (APND; CYP2C6), aniline 4‐hydroxylase (A4H; CYP2E1), nitrosodimethylamine N‐demethylase (NDMA‐ND; CYP2E1) erythromycin N‐demethylase (ERND; CYP3A1) CYP2D1/2 and glutathione S‐transferase (GST). RT‐PCR data and western blotting studies clearly demonstrated that CYP2C6 and CYP2E1 mRNA levels were substantially increased after Urtica treatment, while the level of CYP3A1 mRNA decreased and that of CYP2D1/2 remained unchanged. Urtica treatment significantly induced GST activity in the liver, lung and kidney (66‐, 46‐ and 31‐fold, respectively) while decreasing that of APND (35‐, 61‐ and 94‐fold) and NDMA‐ND (23, 28 and 54‐fold). ERND activity in liver was reduced 45‐fold, but increased in the lung and kidney (78‐ and 144‐fold) after Urtica treatment. These results indicate that Urtica seed extract may have the potential to inhibit and/or induce the metabolism of certain co‐administered drugs. Copyright © 2009 John Wiley & Sons, Ltd.     

Impact of Tetrahydropalmatine on the Pharmacokinetics of Probe Drugs for CYP1A2, 2D6 and 3A Isoenzymes in Beagle Dogs

Tetrahydropalmatine (Tet) exhibit multiple pharmacological activities and is used frequently by clinical practitioners. In this study, we evaluate the in vivo effects of single and repeated oral Tet administrations on CYP1A2, 2D6 and 3A activities in six beagle dogs in a randomized, controlled, open‐label, crossover study. A cocktail approach, with dosages of the probe drugs caffeine (3.0 mg/kg), metoprolol (2.33 mg/kg) and midazolam (0.45 mg/kg), was used to measure cytochrome P450 (CYP) metabolic activities. The cocktail was administered orally as a single dose (12 mg/kg) 1 day prior to and 4 days after repeated oral Tet administrations (12 mg/kg three times daily). The probe drugs and their metabolites in plasma were quantified simultaneously by a validated HPLC technique, and non‐compartmental parameters were used to evaluate metabolic variables for assessment of CYP inhibition or induction. Tet had no or minor impact on the pharmacokinetics and metabolism of the probe drugs caffeine and metoprolol, CYP1A2 and CYP2D6 substrates, respectively. However, Tet increased AUC_{0–24 h} and decreased AUC_{ratio(0–24 h)} (1‐hydroxymidazolam/midazolam ratio) for midazolam statistically significant, both in single or multiple dosing of Tet, with up to 39 or 57% increase for AUC_{0–24 h} and 29% or 22 decrease for AUC_{ratio(0–24 h),} respectively, in line with previous in vitro findings for its CYP3A4 inhibition. The extensive use of Tet and herbal medicines containing Tet makes Tet a candidate for further evaluation of CYP3A‐mediated herb–drug interactions. Copyright © 2016 John Wiley & Sons, Ltd.