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Quantification of Etodolac in Human Plasma for Pharmacokinetics and Bioequivalence Studies in 27 Korean Subjects. 27名韩国人血浆中依托度酸的定量药代动力学和生物等效性研究。
Pub Date : 2017-01-01 DOI: 10.2174/1872312811666170116151004
Il-Dong Song, Ju‐Seop Kang, Hyunjin Kim, Semi Kim, Dongxue Zhao, Shin-Hee Kim, M. Chun, Kyu-Hyun Lee
OBJECTIVE We developed a simple and validated liquid chromatography tandem mass spectrometry( LC-MS/MS) for quantification of etodolac using pioglitazone as an internal standard (IS) to assess pharmacokinetics and to appraise bioequivalence of two formulations of etodolac (reference and tested) in 27 healthy Korean subjects. METHODS Isocratic mobile phase consisted of 10 mM ammonium formate and acetonitrile were used to separate the analytes on a Gemini C18 column. Also, analytes were analyzed by MS/MS in multiple reaction monitoring (MRM) mode using the transitions of (M+H)+ ions, m/z 288.2→ 172.3 and m/z 357.1→ 134.2 for quantification of etodolac and IS each. The standard calibration curves displayed significant linearity within the range of 0.2-30.0 μ g/mL (r2=0.9956, 1/x2 weighting) with LLOQ of 0.1 μg/mL. RESULTS The retention times of etodolac and the IS were 0.77 min and 0.57 min each, indicating the high-throughput potential of the proposed method. The pharmacokinetic parameters were calculated from the plasma samples and data form the reference and test drugs were represented as follows; Area under plasma concentration-time curve (AUCt) (78.03 vs. 84.00 μgxh/mL), AUC∞ (86.67 vs. 93.92 μgxh/mL), maximal plasma concentration (Cmax) (19.49 vs. 18.94 μg/mL), time for maximal concentrations (Tmax) (2.13 vs. 2.26 h), Plasma elimination half-life (T1/2) (8.12 vs. 8.47 h), elimination rate constant (λz) (0.0853 vs. 0.0818 h-1). Pharmacokinetic parameters with 90% confidence interval fall within the bioequivalence range of 80-125%. CONCLUSION Thus, the new testified method was successfully applied for the pharmacokinetic and bioequivalence studies for two etodolac formulations.
目的以吡格列酮为内标(IS),建立一种简单有效的液相色谱-串联质谱(LC-MS/MS)定量方法,评价两种制剂(参比制剂和被试制剂)在27名健康韩国受试者体内的药代动力学和生物等效性。方法以10mm甲酸铵和乙腈为流动相,采用Gemini C18色谱柱分离。利用(M+H)+离子的跃迁,M /z 288.2→172.3和M /z 357.1→134.2,采用多反应监测(MRM)模式对分析物进行质谱分析,分别定量乙酸乙酯和IS。在0.2 ~ 30.0 μg/mL范围内线性良好(r2=0.9956, 1/x2加权),定量限为0.1 μg/mL。结果依托度酸和IS的保留时间分别为0.77 min和0.57 min,表明该方法具有较高的通量潜力。根据血浆样品计算药代动力学参数,参比药和试验药数据如下:血浆浓度-时间曲线下面积(AUCt) (78.03 vs. 84.00 μgxh/mL)、AUC∞(86.67 vs. 93.92 μgxh/mL)、最大血浆浓度(Cmax) (19.49 vs. 18.94 μgxh)、最大浓度时间(Tmax) (2.13 vs. 2.26 h)、血浆消除半衰期(T1/2) (8.12 vs. 8.47 h)、消除速率常数(λz) (0.0853 vs. 0.0818 h-1)。90%置信区间的药代动力学参数在80-125%的生物等效性范围内。结论该方法可用于两种依托度酸制剂的药动学和生物等效性研究。
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引用次数: 0
Pharmacokinetics, Metabolism and Disposition of [14C]XQ-1H After Intravenous Administration to Male Rats. [14C]XQ-1H在雄性大鼠静脉给药后的药代动力学、代谢和处置。
Pub Date : 2017-01-01 DOI: 10.2174/1872312811666170118162931
Yin-lin Qin, Tao Chen, Qiu Jin, Kai Guo, Hao Feng, Dennis Heller, Zhe-ming Gu
OBJECTIVE This study describes the in vivo pharmacokinetics and metabolism of [14C]labeled XQ-1H in male rats. METHODS XQ-1H is a methanesulfonate of XQ, 10-O-(N,N-dimethylaminoethyl)-ginkgolide B, a derivative of ginkgolide B (GB) with enhanced water solubility. Since it is very difficult to synthesize radiolabeled GB, the results obtained in this study may provide helpful insight to further ADME investigation of GB and its analogue compounds. After an i.v. administration of [14C]XQ-1H to male rats, XQ (the freebase form of XQ-1H) was extensively hydrolyzed, moderately metabolized, and mainly excreted in feces (71.5% of the dose) via the biliary route. RESULTS The main enzyme mediated metabolic pathways were mono- and di-demthylation. Using the radiolabel form of XQ-1H, the temporal binding of XQ to red blood cells was observed. CONCLUSION Binding of XQ to RBCs may lower the blood's viscosity and thus provide symptomatic improvement of ischemic stroke patients.
目的研究[14C]标记的XQ-1H在雄性大鼠体内的药动学和代谢。方法sxq - 1h为XQ, 10-O-(N,N-二甲氨基乙基)银杏内酯B的甲磺酸盐,是银杏内酯B (GB)的衍生物,具有较强的水溶性。由于放射性标记GB的合成非常困难,本研究的结果可能为进一步研究GB及其类似物的ADME提供有益的见解。雄性大鼠经静脉给药[14C]XQ- 1h后,XQ (XQ- 1h的自由碱基形式)被广泛水解,适度代谢,主要通过胆汁途径随粪便排出(占剂量的71.5%)。结果酶介导的主要代谢途径为单去甲基化和二去甲基化。利用XQ- 1h的放射性标记形式,观察XQ与红细胞的时间结合。结论XQ与红细胞结合可降低血黏度,改善缺血性脑卒中患者的症状。
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引用次数: 3
A Pharmacokinetic-Pharmacodynamic Model of Tamoxifen and Endoxifen to Predict Their Distribution and Effects on Inhibition of Tumor Growth. 他莫昔芬和恩多西芬的药代动力学-药效学模型预测其分布和抑制肿瘤生长的作用。
Pub Date : 2017-01-01 DOI: 10.2174/1872312811666170815160751
Shengyue Yuan, Qingrong Sun, Yao Chen, Jun Liao

Background: Tamoxifen is widely used in the therapy for breast cancer and has three major metabolites, N-desmethyltamoxifen, 4-hydroxytamoxifen, and endoxifen. Endoxifen has played a major role in the inhibition of tumor growth of breast cancer and the tumor growth is related to endoxifen concentration.

Objectives: The aim of this study was to develop a pharmacokinetic-pharmacodynamic model to predict the distribution of tamoxifen and endoxifen quantitatively, and to discover the anti-tumor effect patterns of tamoxifen and endoxifen.

Methods: The pharmacokinetic-pharmacodynamic model was established by integrating a four compartments pharmacokinetics model and a pharmacodynamic model, the first one include central compartment and peripheral compartment both of which contain tamoxifen and endoxifen. The parameters of the model were calculated by the values of plasma concentrations and the tumor growth data before and after the administration of tamoxifen.

Results: The transport rate k42 (6.0003) of endoxifen from the peripheral compartment to the central compartment and the metabolism rate k34 (0.0031) from tamoxifen to endoxifen in the peripheral compartment were proven to be significant, which showed that tamoxifen and endoxifen are mainly distributed in the central compartment. The model provided reasonable predictions of tumor growth, which was inhibited after the administration and varies with the concentration of endoxifen.

Conclusion: We established a PK-PD model of tamoxifen and endoxifen to predict the tumor growth. The parameters of the pharmacodynamic model, which characterized the tumor growth, revealed the patterns of tamoxifen's anti-tumor functions. The PK-PD model successfully provided illustration for the pharmacokinetics of tamoxifen and endoxifen, and predicted the inhibition effect of endoxifen on the tumor growth.

背景:他莫昔芬广泛应用于乳腺癌的治疗,其主要代谢物有n -去甲基他莫昔芬、4-羟他莫昔芬和内多西芬三种。Endoxifen在抑制乳腺癌肿瘤生长方面发挥了重要作用,肿瘤生长与Endoxifen浓度有关。目的:建立药动学-药效学模型,定量预测他莫昔芬和恩多西芬的分布,探讨他莫昔芬和恩多西芬的抗肿瘤作用规律。方法:采用四室药动学模型和药效学模型相结合的方法建立药动学-药效学模型,四室药动学模型包括中央室和外周室,均含有他莫昔芬和内多西芬。通过给药前后血药浓度和肿瘤生长数据计算模型参数。结果:外周腔室内endoxifen向中央腔室的转运率k42(6.0003)和他莫昔芬向外周腔室内endoxifen的代谢率k34(0.0031)均显著,说明他莫昔芬和endoxifen主要分布在中央腔室。该模型对肿瘤生长预测合理,给药后肿瘤生长受到抑制,且随内毒素浓度的变化而变化。结论:建立了他莫昔芬与内多西芬的PK-PD预测模型。表征肿瘤生长的药效学模型参数揭示了他莫昔芬抗肿瘤功能的规律。PK-PD模型成功地说明了他莫昔芬和内多西芬的药代动力学,并预测了内多西芬对肿瘤生长的抑制作用。
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引用次数: 3
Strong Induction of Cytochrome P450 1A/3A, But not P450 2B, in Cultured Hepatocytes from Common Marmosets and Cynomolgus Monkeys by Typical Human P450 Inducing Agents. 典型人类P450诱导剂对普通狨猴和食蟹猴肝细胞中细胞色素P450 1A/3A的诱导作用较强,但对P450 2B的诱导作用不明显。
Pub Date : 2017-01-01 DOI: 10.2174/1872312810666161114144412
Shotaro Uehara, Y. Uno, Takako Suzuki, Takashi Inoue, M. Utoh, E. Sasaki, H. Yamazaki
BACKGROUND Common marmosets (Callithrix jacchus) and cynomolgus monkeys (Macaca fascicularis) are used as non-human primate models in preclinical studies for drug development. OBJECTIVE The assessment of P450 induction in hepatocytes from marmosets and cynomolgus monkeys was performed using typical P450 inducers. METHODS Induction of cytochrome P450 1-4 family enzymes was analyzed in two lots of cultured hepatocytes from common marmosets and cynomolgus monkeys after 24-h treatment with typical human P450 inducing agents by real-time reverse transcription-polymerase chain reaction. RESULTS Marmoset P450 3A4 mRNA and P450 2C8/2C19 mRNA in hepatocytes were strongly (>10- fold) and weakly (>2) induced by rifampicin, respectively. Marmoset 1A1 and 1A2 mRNA were induced strongly (>200-fold) by β-naphthoflavone and omeprazole. Marmoset P450 2B6 mRNA was induced (~5-fold) by a constitutive androstane receptor agonist, but not by phenobarbital. Cynomolgus monkey P450 3A4 mRNA and P450 1A1 mRNA in cultured hepatocytes were also induced by rifampicin and omeprazole, respectively, but P450 2B6 mRNA was not induced by phenobarbital. CONCLUSION These results indicate that P450 1A/3A induction by typical human P450 inducers in hepatocytes from marmosets and/or cynomolgus monkeys are similar to those of humans (except for P450 2B induction by phenobarbital in humans), suggesting that marmosets and cynomolgus monkeys might be suitable models for evaluating the drug interactions in preclinical studies.
普通狨猴(Callithrix jacchus)和食蟹猴(Macaca fascicularis)被用作非人类灵长类动物模型,用于药物开发的临床前研究。目的采用典型P450诱导剂对狨猴和食蟹猴肝细胞P450的诱导作用进行研究。方法采用实时逆转录-聚合酶链反应法,对普通狨猴和食蟹猴肝细胞经典型人P450诱导剂作用24 h后,细胞色素P450 1-4家族酶的诱导情况进行分析。结果利福平对肝细胞中smarmoset P450 3A4 mRNA和P450 2C8/2C19 mRNA的诱导作用分别为强(>0倍)和弱(>2)。β-萘黄酮和奥美拉唑对绒猴1A1和1A2 mRNA的诱导作用较强,达到200倍。狨猴P450 2B6 mRNA可被组成型雄甾受体激动剂诱导(约5倍),而苯巴比妥则不能。利福平和奥美拉唑均能诱导食蟹猴肝细胞P450 3A4 mRNA和P450 1A1 mRNA表达,而苯巴比妥对P450 2B6 mRNA表达无诱导作用。结论典型人P450诱导剂对狨猴和食蟹猴肝细胞P450 1A/3A的诱导作用与人相似(人类苯巴比妥对P450 2B的诱导作用除外),提示在临床前研究中,狨猴和食蟹猴可能是评估药物相互作用的合适模型。
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引用次数: 12
Ortho-Methylarylamines as Time-Dependent Inhibitors of Cytochrome P450 1A1 Enzyme. 邻甲基芳胺作为细胞色素 P450 1A1 酶的时间依赖性抑制剂
Pub Date : 2017-01-01 DOI: 10.2174/1872312810666161220155226
Jayalakshmi Sridhar, Jiawang Liu, Rajesh Komati, Richard Schroeder, Quan Jiang, Phan Tram, Kevin Riley, Maryam Foroozesh

Background: Members of the cytochrome P450 1A family metabolize many procarcinogens such as polycyclicaromatic hydrocarbons and heterocyclic amines. Inactivation of these enzymes is a prerequisite for cancer prevention and treatment in certain cases. Mechanism-based inhibition (time and co-factor dependent) is an effective method for the inactivation of these enzymes. Our recent study on emodin analogs revealed an anthraquinone with ortho-methylarylamine moiety that exhibited timedependent inhibition of P450 enzymes 1A1 and 1A2.

Methods: To determine whether the amino group or the methyl group or both were responsible for the time-dependent inhibition of these enzymes, a set of eleven compounds containing the orthomethylarylamine moiety were identified through a database search, and studied for the inhibition of the P450 enzymes 1A1, 1A2, 2A6 and 2B1. Our earlier studies on carbazole derivatives provided us with highly selective P450 1A2 inhibitors. Glycine scanning studies were performed on the docked proteinligand complexes of compounds 1-20 in order to understand the contribution of different protein residues towards the ligand binding.

Results: Four compounds were found to cause selective time-dependent inhibition of P450 1A1 with KI values ranging from 0.24 to 8.25 mM. These compounds exhibited only direct inhibition of P450 1A2. Molecular modeling studies of these molecules indicated that the shapes of the molecules, their binding modes, and the methyl substituent in close proximity (4.5-5.7 Å) to the heme-Fe all contributed to their selective time-dependent inhibition activity on P450 1A1. Glycine scanning studies for P450 1A1 indicated that ligand interaction with Phe123 was the strongest binding contributor and similar studies for P450 1A2 indicated that ligand interactions with the phenylalanine residues 226 and 260 were the largest binding contributors.

Conclusion: Four compounds have been identified that exhibit selective time-dependent inhibition of P450 1A1. Modeling studies have indicated that the proximity of the aromatic methyl group to the heme-Fe could be the main contributor for time-dependent inhibition. Future studies will focus on the confirmation of the involvement of the aromatic methyl group in enzyme inactivation.

背景:细胞色素 P450 1A 家族成员可代谢多种致癌物质,如多环芳烃和杂环胺。在某些情况下,使这些酶失活是预防和治疗癌症的先决条件。基于机制的抑制(依赖于时间和辅助因子)是灭活这些酶的有效方法。我们最近对大黄素类似物的研究发现了一种带有邻甲基芳基胺分子的蒽醌,它对 P450 酶 1A1 和 1A2 具有时间依赖性抑制作用:为了确定对这些酶的时间依赖性抑制作用是由氨基、甲基还是两者共同造成的,我们通过数据库搜索确定了一组含有邻甲基芳基胺的 11 种化合物,并研究了它们对 P450 酶 1A1、1A2、2A6 和 2B1 的抑制作用。 我们早先对咔唑衍生物的研究为我们提供了高选择性 P450 1A2 抑制剂。我们对 1-20 号化合物的对接蛋白质配体进行了甘氨酸扫描研究,以了解不同蛋白质残基对配体结合的贡献:结果:发现四种化合物对 P450 1A1 具有选择性时间依赖性抑制作用,KI 值在 0.24 至 8.25 mM 之间。这些化合物只表现出对 P450 1A2 的直接抑制作用。对这些分子进行的分子建模研究表明,分子的形状、结合模式以及靠近血红素-铁(4.5-5.7 Å)的甲基取代基都有助于它们对 P450 1A1 的选择性时间依赖性抑制活性。对 P450 1A1 的甘氨酸扫描研究表明,配体与 Phe123 的相互作用是最强的结合促进因素;对 P450 1A2 的类似研究表明,配体与苯丙氨酸残基 226 和 260 的相互作用是最大的结合促进因素:结论:已发现四种化合物对 P450 1A1 具有选择性时间依赖性抑制作用。建模研究表明,芳香族甲基与血红素-铁的接近可能是时间依赖性抑制的主要因素。今后的研究将侧重于证实芳香族甲基参与酶失活。
{"title":"Ortho-Methylarylamines as Time-Dependent Inhibitors of Cytochrome P450 1A1 Enzyme.","authors":"Jayalakshmi Sridhar, Jiawang Liu, Rajesh Komati, Richard Schroeder, Quan Jiang, Phan Tram, Kevin Riley, Maryam Foroozesh","doi":"10.2174/1872312810666161220155226","DOIUrl":"10.2174/1872312810666161220155226","url":null,"abstract":"<p><strong>Background: </strong>Members of the cytochrome P450 1A family metabolize many procarcinogens such as polycyclicaromatic hydrocarbons and heterocyclic amines. Inactivation of these enzymes is a prerequisite for cancer prevention and treatment in certain cases. Mechanism-based inhibition (time and co-factor dependent) is an effective method for the inactivation of these enzymes. Our recent study on emodin analogs revealed an anthraquinone with ortho-methylarylamine moiety that exhibited timedependent inhibition of P450 enzymes 1A1 and 1A2.</p><p><strong>Methods: </strong>To determine whether the amino group or the methyl group or both were responsible for the time-dependent inhibition of these enzymes, a set of eleven compounds containing the orthomethylarylamine moiety were identified through a database search, and studied for the inhibition of the P450 enzymes 1A1, 1A2, 2A6 and 2B1. Our earlier studies on carbazole derivatives provided us with highly selective P450 1A2 inhibitors. Glycine scanning studies were performed on the docked proteinligand complexes of compounds 1-20 in order to understand the contribution of different protein residues towards the ligand binding.</p><p><strong>Results: </strong>Four compounds were found to cause selective time-dependent inhibition of P450 1A1 with KI values ranging from 0.24 to 8.25 mM. These compounds exhibited only direct inhibition of P450 1A2. Molecular modeling studies of these molecules indicated that the shapes of the molecules, their binding modes, and the methyl substituent in close proximity (4.5-5.7 Å) to the heme-Fe all contributed to their selective time-dependent inhibition activity on P450 1A1. Glycine scanning studies for P450 1A1 indicated that ligand interaction with Phe123 was the strongest binding contributor and similar studies for P450 1A2 indicated that ligand interactions with the phenylalanine residues 226 and 260 were the largest binding contributors.</p><p><strong>Conclusion: </strong>Four compounds have been identified that exhibit selective time-dependent inhibition of P450 1A1. Modeling studies have indicated that the proximity of the aromatic methyl group to the heme-Fe could be the main contributor for time-dependent inhibition. Future studies will focus on the confirmation of the involvement of the aromatic methyl group in enzyme inactivation.</p>","PeriodicalId":11339,"journal":{"name":"Drug metabolism letters","volume":"10 4 1","pages":"270-277"},"PeriodicalIF":0.0,"publicationDate":"2017-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6697106/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68049814","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Analysis of Elevated Levels of Nandrolone Decanoate Induced Cytochrome- P450 Alterations in Mice. 十酸诺龙高水平诱导小鼠细胞色素P450改变的分析。
Pub Date : 2017-01-01 DOI: 10.2174/1872312811666171114145535
Parmita Chowdhury, Rita Mahanta

Background: Frequent recreational use of Anabolic Androgenic Steroids (AAS) is an instance of substance abuse which mimics the status of a natural hormone and upon prolonged exposure may lead to adverse drug reactions. These adverse drug reactions proceed in a manner so as to alter the normal metabolism of an enzyme mediated pathway such as the Cytochrome P450 (CYP) family of enzymes.

Objective: The present study was conducted to investigate the impact of overuse of Nandrolone Decanoate (ND), an AAS, upon CYP enzyme activity and a CYP gene, belonging to CYP1 family.

Methods: The study was carried out using normal and ND treated male albino mice. Genetic analysis was conducted using normalized and treated cDNA and reverse transcriptase polymerase chain reaction based assays. For enzyme assay, 0.1ml of 25 mg ND was administered to the animals twice a week for a period of 90 days. Genetic analysis was carried out with the same dose but administered for a period of 360 days.

Results: CYP enzyme activity increased significantly (p<0.01) in the ND treated group of animals compared to that in the normal group. However, no noticeable alteration was observed at the molecular level.

Conclusion: From the present study it could be inferred that, at elevated doses, ND has the potential to alter hepatic CYP enzyme activity without any modification in the CYP gene. This could be due to a possible adaptive response of the living system to such drugs.

背景:频繁的娱乐性使用合成代谢雄激素类固醇(AAS)是一种药物滥用的例子,它模仿天然激素的状态,长时间暴露可能导致药物不良反应。这些药物不良反应以某种方式进行,从而改变酶介导途径的正常代谢,如细胞色素P450 (CYP)酶家族。目的:探讨过量使用醋酸诺龙(ND)对CYP酶活性及CYP1家族CYP基因的影响。方法:以正常和ND治疗的雄性白化小鼠为研究对象。采用归一化和处理过的cDNA和逆转录聚合酶链式反应进行遗传分析。酶测定,每周两次给药0.1ml 25 mg ND,连续90天。遗传分析以相同剂量进行,但给药期为360天。结果:CYP酶活性显著增加(p结论:从本研究可以推断,在高剂量下,ND有可能改变肝脏CYP酶活性,而不改变CYP基因。这可能是由于生命系统对这些药物的一种可能的适应性反应。
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引用次数: 0
Evaluation of Farnesoid X Receptor Target Gene Induction in Human Hepatocytes: Amino Acid Conjugation. 法脂类X受体靶基因在人肝细胞诱导的评价:氨基酸偶联。
Pub Date : 2017-01-01 DOI: 10.2174/1872312812666171227213946
Jean Simmermacher, Michael Sinz

Background: The nuclear hormone receptor, Farnesoid X Receptor (FXR) regulates the transcription of genes associated with bile acid metabolism and disposition.

Objective: This study investigates possible changes in the expression of target genes responsible for amino acid conjugation, i.e., Bile Acid-CoA Synthetase (BACS) and bile acid-CoA: amino acid Nacetyltransferase (BAT). These genes have been shown to be inducible by FXR agonists in rat models, however, to date no studies have been conducted in a human hepatocyte model.

Results: In human hepatocytes, treatment with the FXR agonists GW4064 (1.0 µM) and WAY362450 (0.1 µM) did not significantly induce the mRNA expression of BACS and BAT genes. However, other target genes associated with FXR activation, such as Bile Salt Export Pump (BSEP), Short Heterodimer Partner (SHP), Multidrug Resistance-associated Protein 2 (MRP2) and Multidrug Resistance Protein 3 (MDR3), were upregulated. Interestingly, a follow up study conducted in rat hepatocytes indicated that GW4064 induced the BACS gene while WAY362450 induced the BAT gene, confirming literature results that these genes can be induced in rat.

Conclusion: In conclusion, there appears to be some species differences in the activation of FXR target genes.

背景:核激素受体Farnesoid X receptor (FXR)调节与胆汁酸代谢和处置相关基因的转录。目的:探讨氨基酸偶联靶基因,即胆汁酸-辅酶a合成酶(BACS)和胆汁酸-辅酶a:氨基酸乙酰转移酶(BAT)表达的可能变化。这些基因已在大鼠模型中被FXR激动剂诱导,然而,迄今为止尚未在人肝细胞模型中进行研究。结果:在人肝细胞中,FXR激动剂GW4064(1.0µM)和WAY362450(0.1µM)未显著诱导BACS和BAT基因的mRNA表达。然而,与FXR激活相关的其他靶基因,如胆汁盐输出泵(BSEP)、短异二聚体伴侣(SHP)、多药耐药相关蛋白2 (MRP2)和多药耐药蛋白3 (MDR3)上调。有趣的是,后续在大鼠肝细胞中进行的研究表明,GW4064诱导BACS基因,WAY362450诱导BAT基因,证实了文献中这些基因在大鼠体内可以诱导的结果。结论:综上所述,FXR靶基因的激活存在一定的物种差异。
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引用次数: 3
Therapeutic Potentials and Cytochrome P450-Mediated Interactions Involving Herbal Products Indicated for Diabetes Mellitus. 中药产品对糖尿病的治疗潜力和细胞色素p450介导的相互作用
Pub Date : 2017-01-01 DOI: 10.2174/1872312811666171121104431
Cyprian O Onyeji, Sharon I Igbinoba, Gbola Olayiwola

Background: There has been an increase in the use of herbal products to complement conventional drugs in the treatment of various diseases especially in developing countries. This may be attributable to the potential cost-effectiveness and ease of accessibility of these products as well as the perception of their safety profiles. However, there are numerous literature reports on herbs altering the pharmacokinetics and pharmacodynamics of other co-administered drugs thereby modulating the therapeutic outcomes. The prevalence of diabetes is on a steady increase worldwide and it is now identified as one of the main threats to human health.

Objective: It is important that knowledge on specific effects of antidiabetic herbs and their products on drug metabolizing enzymes are updated and documented so as to ensure optimization of their therapeutic utility. This review, therefore, aims to highlight herbal products with evidence-based antidiabetic effects, identify their bioactive phyto-constituents, and also focus on the important Cytochrome P450 and consequences of their inhibition or induction.

Methods: An extensive literature search was undertaken and the information obtained were critically analyzed and discussed.

Results and conclusion: The literature abounds with reports on the utilization of herbal medications for the treatment of diabetes mellitus since time immemorial, but very few of these herbal products have undergone clinical trials. Also, studies on the herb-drug interactions were limited. Due to the complex phytochemical composition of the herbs, concomitant administration with conventional drugs resulted in alterations of pharmacological effects of some drugs. Evidences of beneficial interactions were identified for medical exploitation.

背景:特别是在发展中国家,越来越多地使用草药产品来补充传统药物治疗各种疾病。这可能是由于这些产品的潜在成本效益和易于获取以及对其安全概况的认识。然而,有大量的文献报道,草药改变药代动力学和药效学的其他共同给药的药物,从而调节治疗结果。糖尿病的患病率在世界范围内稳步上升,现已被确定为对人类健康的主要威胁之一。目的:更新和记录抗糖尿病中药及其制品对药物代谢酶的特异性作用,以确保其治疗效果的优化。因此,本综述旨在重点介绍具有循证抗糖尿病作用的草药产品,鉴定其生物活性植物成分,并关注重要的细胞色素P450及其抑制或诱导的后果。方法:进行广泛的文献检索,并对所获得的信息进行批判性分析和讨论。结果与结论:自古以来,文献中就有大量关于利用中药治疗糖尿病的报道,但这些中药产品很少进行临床试验。此外,对中药相互作用的研究也很有限。由于中草药的植物化学成分复杂,与常规药物合用会导致某些药物的药理作用发生改变。在医学开发中发现了有益相互作用的证据。
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引用次数: 8
A Novel Liquid Chromatography Tandem Mass Spectrometry Method for the Estimation of Bilirubin Glucuronides and its Application to In Vitro Enzyme Assays. 一种新的液相色谱串联质谱法测定胆红素葡萄糖醛酸酯及其在体外酶分析中的应用。
Pub Date : 2017-01-01 DOI: 10.2174/1872312810666161124143522
S. P. Putluru, M. Matta, Deepak Ahire, Murali Subramanian, M. Sinz, S. Mandlekar
BACKGROUND Bilirubin is a toxic waste product of metabolism, eliminated mainly through UGT1A1 mediated conjugation to mono- and di-glucuronides. Due to the potentially low Km value of bilirubin glucuronidation, the quantitative sensitivity obtained with most UV/visible light detection methods are not sufficient to accurately calculate UGT1A1 enzyme kinetics at low bilirubin concentrations. In addition, bilirubin, as well as its metabolites, are unstable during sample preparation and bioanalysis. This necessitates the need for a rapid, sensitive and robust assay to measure bilirubin glucuronides. METHODS A robust LC-MS/MS method was developed to measure low levels of bilirubin glucuronides accurately from in vitro incubations, as well as stabilizing the analytes during sample preparation and analysis. The metabolites were quantified using a qualitative/quantitative approach utilizing UV to MS correction, thereby eliminating the need for synthetic standards. RESULTS The method was sensitive enough to quantify mono- and di-glucuronides as low as 3 nM from in vitro incubations, and kinetic data was determined for total glucuronide formation. The Km and Vmax values for total bilirubin glucuronide formations were determined to be 0.05 ± 0.01 μM and 181.9 ± 5.3 pmol/min/mg-protein, respectively, in human recombinant UGT1A1, and 0.23 ± 0.05 μM and 875 ± 45 pmol/min/mg protein in human liver microsomes (HLM). CONCLUSION We have developed a sensitive LC-MS/MS based method for the quantitation of bilirubin and its glucuronides from in vitro incubations. This method was successfully utilized to determine bilirubin glucuronidation kinetics in HLM and human rUGT1A1.
胆红素是一种有毒的代谢废物,主要通过UGT1A1介导的单和双葡萄糖醛酸盐偶联来消除。由于胆红素糖醛酸化的Km值可能较低,大多数紫外/可见光检测方法获得的定量灵敏度不足以准确计算低胆红素浓度下UGT1A1酶的动力学。此外,胆红素及其代谢物在样品制备和生物分析过程中是不稳定的。这就需要一种快速、灵敏和可靠的测定胆红素葡萄糖醛酸苷的方法。方法建立了一种高效液相色谱-质谱联用(LC-MS/MS)方法,可准确测定体外培养物中低水平胆红素葡萄糖醛酸酯,并在样品制备和分析过程中稳定分析物。代谢物采用定性/定量方法进行定量,利用紫外-质谱校正,从而消除了合成标准品的需要。结果该方法灵敏度高,可在体外培养3 nM范围内定量测定单、双葡萄糖苷,并测定了总葡萄糖苷生成的动力学数据。在重组人UGT1A1中,总胆红素葡萄糖醛酸形成的Km和Vmax分别为0.05±0.01 μM和181.9±5.3 pmol/min/mg-protein;在人肝微粒体(HLM)中,总胆红素葡萄糖醛酸形成的Km和Vmax分别为0.23±0.05 μM和875±45 pmol/min/mg-protein。结论建立了一种高效液相色谱-质谱联用技术,可用于体外培养物中胆红素及其葡糖醛酸酯的定量分析。该方法成功地用于测定HLM和人rUGT1A1中胆红素糖醛酸化动力学。
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引用次数: 13
Genotoxicity of 4-(piperazin-1-yl)-8-(trifluoromethyl)pyrido[2,3-e][1,2,4] triazolo[4,3-a]pyrazine, a Potent H4 Receptor Antagonist for the Treatment of Allergy: Evidence of Glyoxal Intermediate Involvement. 4-(哌嗪-1-酰基)-8-(三氟甲基)吡啶[2,3-e][1,2,4]三唑啉[4,3-a]吡嗪是治疗过敏的有效H4受体拮抗剂:乙醛中间体参与的证据
Pub Date : 2017-01-01 DOI: 10.2174/1872312811666171106151730
Mithat Gunduz, Amanda L Cirello, Peter Klimko, Jennifer L Dumouchel, Upendra A Argikar

Background: 4-(piperazin-1-yl)-8-(trifluoromethyl)pyrido[2,3-e][1,2,4]triazolo[4,3-a]pyrazine (1) is a small-molecule which demonstrated a sub-nM inhibitory potency toward the histamine H4 receptor (H4R). However, it was found to be mutagenic in an in vitro Ames assay. Metabolic bioactivation of 1 could potentially arise from the piperazine moiety by forming reactive intermediates such as glyoxal, aldehyde-imine and/or iminium ion, which could all lead to genotoxicity. The aim of this study was to investigate bioactivation of 1 to determine the potential causes of the genotoxicity and mitigate liabilities in this scaffold.

Methods: 1 was investigated for its genotoxicity in phenobarbital and β-naphthoflavone induced Sprague Dawley rat liver S9 fractions. Trapping agents such as o-phenylenediamine was used postincubation.

Results: Following metabolic profiling of 1, two oxidative metabolites were observed and identified in phenobarbital- and β -naphthoflavone induced Sprague Dawley rat liver S9 fractions. Metabolic pathway of 1 was primarily mediated by the metabolism of the piperazine moiety. The trapped glyoxal was identified by using high resolution LC-MS instrument. Structural characterization of the trapped glyoxal was determined by comparison of retention time, accurate mass measurement and Collision Induced Dissociation (CID) spectra to authentic standard.

Conclusion: In the present investigation, a novel method was developed to trap glyoxal, which may potentially be liberated from piperazine moiety. These findings led to modifications on the piperazine ring to mitigate the bioactivation pathways leading to mutagenicity. Subsequently, the next generation compounds with modified piperazine moiety, retained H4R inhibitory potency in vitro and were not genotoxic in the Ames mutagenicity assay.

背景:4-(哌嗪-1-酰基)-8-(三氟甲基)吡嗪[2,3-e][1,2,4]三唑罗[4,3-a]吡嗪(1)是一种对组胺H4受体(H4R)具有亚nm抑制作用的小分子。然而,在体外Ames实验中发现它具有诱变性。1的代谢生物激活可能来自哌嗪部分,形成活性中间体,如乙二醛、醛亚胺和/或亚胺离子,这些都可能导致遗传毒性。本研究的目的是研究1的生物活性,以确定遗传毒性的潜在原因,并减轻该支架的责任。方法:研究其对苯巴比妥和β-萘黄酮诱导大鼠肝脏S9部位的遗传毒性。孵育后使用邻苯二胺等诱捕剂。结果:在苯巴比妥和β -萘黄酮诱导的Sprague Dawley大鼠肝脏S9组分中,观察并鉴定了2种氧化代谢物。1的代谢途径主要由哌嗪部分的代谢介导。采用高分辨率LC-MS对捕获的乙二醛进行鉴定。通过保留时间、精确质量测量和碰撞诱导解离(CID)光谱与真实标准相比较,确定了捕获的乙二醛的结构特征。结论:本研究建立了一种新的方法来捕获可能从哌嗪部分释放的乙二醛。这些发现导致对哌嗪环进行修饰,以减轻导致致突变性的生物激活途径。随后,具有修饰哌嗪部分的下一代化合物在体外保留了H4R抑制效力,并且在Ames诱变试验中没有遗传毒性。
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引用次数: 3
期刊
Drug metabolism letters
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