对乙酰氨基酚羧酸酯结构和在人羧酸酯酶的代谢稳定性的关系

Relationship of structure and metabolic stabilities of paracetamol derived carboxylic esters in recombinant human carboxylesteras

Metabolite profiling of 4-methylescultin in rat using ultra-fast liquid chromatography combined with tandem mass spectrometry

Metabolite profiling of 4-methylescultin in rat using ultra-fast liquid chromatography combined with tandem mass spectrometr

一类具有抗肿瘤活性的新型紫杉烷卤代衍生物

一类具有抗肿瘤活性的新型紫杉烷卤代衍生物，其特征在于：所述为具有如结构通式的新型紫杉烷卤代衍生物及以其作为活性成分的抗肿瘤药物如式I，其中R为H或Ac；R’具有下右式结构，其中R”为选自-OMe，-OH的取代基，X为选自Cl，Br中的卤素。本发明中所示的化合物对于白血病细胞(K562)，非小细胞肺癌细胞(A549)，结肠癌细胞 (HT-29)，卵巢癌细胞(OVCAR-3)，人成骨肉瘤细胞(Saos-2)，乳腺癌细胞 (MCF-7)等多种肿瘤细胞系均表现出与紫杉醇相似的细胞毒性，具有作为抗癌药物开发的价值。带填

基于超快速液相色谱/二极管阵裂管检测器采用盒式分析检测五个CYP探针底物及其代谢产物

A valid, efficient and time-saving cassette analytical method has been developed for simultaneous determination of five CYP probe drugs and their metabolites by using ultra-fast liquid chromatography coupled with diode array detector (UFLC-DAD). Chromatographic separation was performed with a Shim-pack XR-ODS column (75 mm × 2.0 mm, 2.2 µ), with the mobile phase consisted of methanol and 0.5% formic acid at the flow rate of 0.4 mL min-1. Five probe reactions for CYP isoforms including Phenacetin-O-deethylation (CYP1A2), Coumarin-7-hydroxylation (CYP2A6), Paclitaxel-6-hydroxylation (CYP2C8), Chlorzoxazone-6-hydroxylation (CYP2E1), Nifedipine oxidation (CYP3A4) were selected for this study. The detection wavelengths of five CYP probe reactions were set at the maximal absorbent wavelengths of Phenacetin, Coumarin, Paclitaxel, Chlorzoxazone and Nifedipine, were 245 nm, 320 nm, 230 nm, 280 nm and 250 nm, respectively. The limit of detection of Phenacetin, Coumarin, Paclitaxel, Chlorzoxazone, Nifedipine and their metabolites ranged from 0.08 to 0.25 ng. The linear detection range of Phenacetin, Coumarin, Paclitaxel, Chlorzoxazone, Nifedipine and their metabolites ranged from 0.8 to 200 ng with good correlation coefficient (>0.9998). The inter-day and intra-day reproducibility were evaluated and the results shown that the relative standard deviations (RSDs) of the retention times and peak areas were less than 2.1 % for the CYP probe drugs and their metabolites. All detected compounds could be isolated with satisfactory resolution within 16 minutes, and the method was successfully applied in rapid assessment and screening of different CYPs activities of human liver microsome from 15 individuals. The five samples for each probe reaction after 30 min incubation with human liver microsome and cofactors were pooled with equal volume, and then analyzed by UFLC-DAD. The conversion of each probe drug was determined to display the activity of the corresponding CYP isoform. The quantitative results from the cassette analysis procedure agreed well with those results obtained from conventional discrete LC analysis. The UFLC-based method offers a significant increase in analytical throughput and can be used in rapid assessment of different CYPs activities of animal liver microsomes from different species and individuals. Furthermore, the method also can be applied in rapid screening of inhibitors of each CYP isoform for drug-drug interaction related studies. The proposed cassette analysis method is rapid, more accurate and more acceptable than previously reported cocktail methods which were less accurate due to the existed competitive inhibitions from several probe substrates

In Vitro Metabolic Study of Boc5 in human tissues and Animal Liver Microsomes

As the first nonpeptidic agonist of glucagon-like peptide 1 receptor, Boc5 can invoke sustained glycemic control and weight loss in diabetic C57BL/6J mice, and has become a promising hit compound for antidiabetic drug. Boc5 displays good efficiency via i.v. administration but loss efficiency via oral administration, thus its ADME properties should be studied. In the present study, the in vitro investigation on metabolic stability of Boc5 was first reported. Both phase I and phase II metabolic studies were carried out by using a series of tissues and microsomes from human and three experimental animals including pig, rat and C57BL/6J mice. Significant species difference in metabolism of Boc5 was presented, Boc5 can not be metabolized by human plasma, liver and intestine microsome, as well as rat and pig liver microsomes. In contrast, Boc5 can be rapid metabolized by C57BL/6J mice liver microsome (~50% conversion at 1 h), and three metabolites (M-1, M-2, and M-3) were detected by using liquid chromatography/mass spectrometry (LC/MS). M-1 was confirmed as 2-thenoic acid, and M-3 and M-2 were assigned as two metabolites after hydrolysis of one and two thenoic acids from Boc5, respectively, based on the retention times and mass spectra by comparison with standards. These results revealed that the hydrolysis of thiophene carboxylic ester bond was the major metabolic pathway of Boc5, indicating esterases invlolved in this biotransformation. Further studies have shown that carboxylesterase in C57BL/6J mice liver display the key role in rapid hydrolysis of Boc5 by using a range of inhibitors. All of these results were very useful to develop the next generation nonpeptidic agonists and to choose suitable experimental animal for whole tests

一种新型细胞色素CYP3A4酶特异性探针反应及其应用

本发明提供了一种新型细胞色素CYP3A4酶的特异性探针反应及其应用：戈米辛A可作为CYP3A4酶的探针底物来检测该酶的活性，以戈米辛A或其药物制剂作为特异性的探针底物，通过在单位时间内定量测定底物戈米辛A的减少速率或产物8-羟基戈米辛A的生成速率作为细胞色素CYP3A4酶活的评价指标。该探针底物还具有高安全性，可作为整体探针使用，通过静脉注射使待测哺乳动物服用0.1至500mg/kg体重的戈米辛A或其药物制剂；在0至24小时内选取时间点，收集待测动物的血浆样本；测定底物戈米辛A的减少速率或产物8-羟基戈米辛A的生成速率，作为整体细胞色素CYP3A4酶活性的评价指标。本发明可实现不同来源的生物样本及体内CYP3A4酶活的定量评估

一种人羧酸酯酶CES2的高特异性荧光探针及其应用

一种人羧酸酯酶CES2的高特异性荧光探针及其应用，该特异性探针底物是3-羟基黄酮类化合物酯类衍生物，其可用于检测不同生物样品中CES2的存在与否及其活力的定量测定。酶活具体测定流程如下：选择3-羟基黄酮酯类衍生物水解反应为探针反应，选择适宜底物浓度在线性反应区间内通过定量检测单位时间内其水解代谢产物3-羟基黄酮的生成量来测定各生物样品、细胞、在体及整体器官中CES2酶的实际活性。本发明不仅可用于不同来源生物样本中CES2酶活的定量评估，此外借助该探针反应还可用于体外快速筛选CES2的抑制剂