儿茶素抗氧化活性的构效关系

儿茶素(catechins)分为酯型儿茶素(EGCG和ECG)和游离型儿茶素(EC和EGC)。酯型儿茶素的抗氧化活性比游离型儿茶素的强[1]。儿茶素类化合物的基本结构均为α 连(或邻)苯酚基苯并吡喃。由于儿茶素分子具有α 连(或邻)苯酚基结构,使它们具有强抗氧化的特性[2]。深入研究儿茶素类抗氧化的构效关系,对天然抗氧化剂的筛选具有重要意义。1　方法　　选用EGCG、ECG、EGC、EC作为模型分子。这四种分子中A环的5 、7 位均带酚羟基,不同的是在C环的2 位带邻苯酚基或连苯三酚基,3 位上带羟基或没食子酸结构。分子的原子编号及结构式见图1,…     

茶中儿茶素的分离分析方法研究进展

儿茶素类是茶叶药效的主要活性组分.天然产生的八大儿茶素为儿茶素(C)、表儿荼素(EC)、没食子儿茶素(GC)、表没食子儿荼素(EGC)、儿茶素没食子酸酯(CG)、表儿荼素没食子酸酯(ECG)、没食子儿荼素没食子酸酯(GCG)、表没食子儿茶素没食子酸酯(EGCG).本文综述了分离分析茶儿荼素的常用有效方法.     

脉冲辐解研究茶多酚4个组分清除羟自由基机理

采用脉冲电子辐解瞬态吸收光谱法 ,在pH 7的磷酸缓冲水溶液中测定茶多酚 4个组分表没食子儿茶素没食子酸酯 (EGCG)、表儿茶素没食子酸酯 (ECG)、表没食子儿茶素 (EGC)和表儿茶素 (EC)的清除羟自由基速率常数分别为 1 .5× 1 0 10 ,1 .4× 1 0 10 ,9.7× 1 0 9和 9.2× 1 0 9dm3 ·mol-1·s-1,并提出了EGCG和ECG在清除羟自由基的过程中存在一个脱没食子酸基团的机理     

半制备型液相色谱法分离纯化茶叶鲜叶中7种儿茶素类化合物

建立了从茶叶鲜叶中分离纯化7种儿茶素类化合物(没食子儿茶素(GC)、表没食子儿茶素(EGC)、儿茶素(C)、表没食子儿茶素没食子酸酯(EGCG)、表儿茶素(EC)、表没食子儿茶素3-O-(3-O-甲基)没食子酸酯(EGCG3"Me)和表儿茶素没食子酸酯(ECG))的半制备色谱法。铁观音鲜叶经甲醇超声浸提、浓缩、氯仿萃取后,向水相中加入碱式醋酸铅沉淀,得到茶多酚粗品。分别以甲醇-水、乙腈-水作为流动相,采用半制备色谱法纯化7种儿茶素类化合物,纯度均达到90%。此外,利用同样的方法分离纯化另外两种茶叶鲜叶中的7种儿茶素类化合物,得到相似的结果。该方法以溶剂提取、离子沉淀结合半制备色谱,适于简单、高效地同时分离制备多种儿茶素类化合物。     

改性木粉填充柱对茶多酚与咖啡因分离应用的研究

用乙基纤维素、己二酸和丙烯酰胺制成浆液改性剂,对木粉进行改性.研究对比了木粉在改性前后对茶多酚(TP)与咖啡因(Caf)的分离效果.研究结果表明,已改性的木粉对TP中3种主要成分(EGC,EGCG和ECG)与Caf的分离效果优于未改性木粉.改性后的木粉可用于TP各组分与Caf分离时的前处理.通过该前处理可将绿茶提取液分离为两部分:一部分主要由咖啡因和表没食子儿茶素(EGC)组成,另一部分主要由表没食子儿茶素没食子酸酯(EGC)和表儿茶素没食子酸酯(ECG)组成.     

茶叶中甲基化儿茶素的分离、纯化和高效液相色谱法分析

以日本Benifuji绿茶为原料,用50%乙腈提取茶叶有效成分,提取液经氯仿脱咖啡碱和色素、乙酸乙酯和HP-20大孔树脂富集儿茶素,得多酚含量高于80%的茶多酚粗品,再经ToyopearHW-40S柱层析分离纯化,得EGC、ECG、EGCG、EC及两种未知成分。经1H-NMR、MS和HPLC等分析,两种未知成分为(-)-表没食子儿茶素3-O(3-O-甲基)没食子酸酯(EGCG3″Me)和(-)-3-O-甲基-表儿茶素没食子酸酯(ECG3′Me)。采用TSKgelODS-100Z色谱柱和二极管阵列检测器(DAD),建立了一种快速、灵敏分析茶叶中儿茶素和甲基化儿茶素的高效液相色谱法。色谱条件:TSKgelODS-100Z色谱柱,流动相为KH2PO4(pH2.5)-甲醇系统,采用梯度洗脱,流速1.0mL/min,柱温40℃,进样量20μL。结果表明,6种儿茶素在一定范围内线性良好,相关系数为0.9996~1.0000,平均加标回收率在90.8%~105.9%之间,相对标准偏差均小于3.5%。     

环氧合酶－2抑制剂的三维定量构效关系研究

建立三环系COX－1和COX－2抑制剂结构与活性的三维定量构效关系模型，为设 计新型的具有选择性的COX－2抑制剂提供线索。通过与酶的对接并优化，确定化合 物在受体结合腔中的构象，利用比较分子力场分析方法建立三维定量构效关系模型 。模型1R_(cv)~2=0.685，最佳主成分数为6，传统相关系数为R~2=0.988, F-726. 2，标准偏差S = 0.080；模型2 R_(cv)~2 = 0.573，最佳主成分数为6，传统相关 系数为R~2=0.996, F = 1147.6，标准偏差S = 0.034。所得的模型不仅解释了化合 物的构效关系，而且对预测集中的化合物有很好的预测能力；比较不同模型的系数 相关图，分析了结构与活性、结构与选择性的关系，得到的结果可以指导新化合物 的设计与合成。     

高效液相色谱-质谱法及模式识别法用于鉴别普洱生茶与熟茶

采用高效液相色谱-质谱联用技术及高效液相色谱法对生熟普洱茶中的主要成分进行定性和定量分析。鉴定出普洱茶水溶液中8种主要成分,分别为没食子酸(GA)、没食子酸儿茶素(GC)、表没食子酸儿茶素(EGC)、儿茶素(C)、咖啡因(CAF)、表儿茶素(EC)、表没食子酸儿茶素没食子酸酯(EGCG)和表儿茶素没食子酸酯(ECG)。以这8种成分的含量为指标,对普洱生茶和熟茶各20批进行主成分分析、聚类分析和判别分析,能准确地区分普洱生茶与熟茶。     

绿茶多酚对自由基诱导的红细胞膜过氧化的抑制作用

采用水溶性偶氮引发剂2,2'-偶氮二(2-脒基丙烷)二盐酸盐(AAPH)在37引发入血红细胞膜的过氧化,通过测定氧气吸收及维生素E的消耗研究了过氧化过程的动力学,并对从绿茶中提取的主要多酚类化合物的抗氧化活性做了定量研究。使用的绿茶多酚有:(-)-表儿茶素(EC),(-)-表儿茶素(EGC),(-)-表儿茶素酸酯(ECG)和(-)-表儿茶素培酸酯(EGCG)。结果表明,这些绿茶多酚能够显著缩短过氧化反应的动力学链长,有效地抑制红细胞膜的过氧化。抗氧化活性顺序为:EC〉GCG〉EGCG〉EGC。     

评估石杉碱甲杂合体衍生物作为AD症药物活性顺序的简单方法

为了合理化设计、指导具有高活性、低毒性的石杉碱甲杂合体(Huprine X)衍生物的合成, 建立了一个计算结合自由能的新方案, 以实现对该类衍生物活性排列顺序的预测. 该结合自由能由三部分组成: 抑制剂和靶酶的相互作用能; 活性位点残基在结合前后的构象绝对自由能的变化; 抑制剂从稳定构象变为活性构象的自由能增加值. 通过计算已合成的14个杂合体衍生物的结合自由能, 结果显示理论值和实验测定的生物活性值有很好的等级相关性, 其斯皮尔曼相关系数为0.85, 证明了该方法的可行性.     

Inhibition of gut bacterial β-glucuronidase by chemical components from black tea: Inhibition interactions and molecular mechanism

Gut bacterial β-glucuronidase is regarded as an important molecular target for several therapeutic applications. Inhibitors of β-glucuronidase can effectively alleviate the drug-induced gastrointestinal tract toxicity. In this study, the ethanol extracts of black tea was found to display significant inhibitory activities against Escherichia coli β-glucuronidase (EcGUS), and seven polyphenols including catechins and theaflavins were identified as the key components responsible for the strong inhibitory potency of black tea towards EcGUS. Among these seven identified naturally occurring inhibitors, (−)-catechin gallate (CG), theaflavin-3-monogallate (TF-3-G), theaflavin-3′-monogallate (TF-3′-G) and theaflavin-3,3′-digallate (TFDG) were more potent inhibitors of EcGUS compared with (−)-epicatechin gallate (ECG), (−)-gallocatechin gallate (GCG), and (−)-epigallocatechin gallate (EGCG). Furthermore, molecular docking and molecular dynamics simulation results further indicated that TFDG could bind in the cavity of EcGUS and interacted with key residues Ser360, Glu413 and Ile560 of EcGUS through hydrogen bonds. Taken together, these data offer important information for efficient development of black tea and its catechins and theaflavins constituents for treating drug-induced enteropathy.     

Evaluation inhibitory activity of catechins on trypsin by capillary electrophoresis–based immobilized enzyme microreactor with chromogenic substrate

In this study, a capillary electrophoresis‐based online immobilized enzyme microreactor was developed for evaluating the inhibitory activity of green tea catechins and tea polyphenol extracts on trypsin. The immobilized trypsin activity and other kinetic parameters were evaluated by measuring the peak area of the hydrolyzate of chromogenic substrate S‐2765. The results indicated that the activity of the immobilized trypsin remained approximately 90.0% of the initial immobilized enzyme activity after 30 runs. The value of Michaelis–Menten constant (K_m) was (0.47 ± 0.08) mM, and the half‐maximal inhibitory concentration (IC₅₀) and inhibition constant (K_i) of benzamidine were measured as 3.34 and 3.00 mM, respectively. Then, the inhibitory activity of four main catechins (epicatechin, epigallocatechin, epicatechin gallate, and epigallocatechin gallate) and three tea polyphenol extracts (green tea, white tea, and black tea) on trypsin were investigated. The results showed that four catechins and three tea polyphenol extracts had potential trypsin inhibitory activity. In addition, molecular docking results illustrated that epigallocatechin gallate, epicatechin gallate, epicatechin, and epigallocatechin were all located not only in the catalytic cavity, but also in the substrate‐binding pocket of trypsin. These results indicated that the developed method is an effective tool for evaluating inhibitory activity of catechins on trypsin.     

Fractionation of Pharmacologically Active Plant Polyphenols by Centrifugal Partition Chromatography

Abstract

Centrifugal partition chromatography (CPC) has been applied to the preparative fractionation of pharmacologically active polyphenols contained in the crude extracts from three medicinal plants. 1) (-)-Epigallocatechin gallate, the main component of tea polyphenols, which inhibits tumor promotion, 2) oligomeric hydrolyzable tannins, having host-mediated anti-tumor activity and antiviral activity, contained in Woodfordia fruticosa flower, 3) polyphenolic components of licorice, having anti-HIV activity. The results obtained with cartridges of medium volume and of small volume were compared during the fractionation of tea polyphenols.     

Green,Black and Rooibos Tea Inhibit Prostaglandin E2 Formation in Human Monocytes by Inhibiting Expression of Enzymes in the Prostaglandin E2 Pathway

The formation of prostaglandin E2 (PGE2) is associated with adverse inflammatory effects. However, long-term treatment with nonsteroidal anti-inflammatory drugs (NSAIDs) comes with risk of severe side effects. Therefore, alternative ways to inhibit PGE2 are warranted. We have investigated the effects of tea extracts and the polyphenols epigallocatechin gallate (EGCG) and quercetin on PGE2 formation, determined by immunoassay, and protein expression, determined by immunoblotting, of cytosolic phospholipase A2 (cPLA2), cyclooxygenase 2 (COX-2) and microsomal PGE synthase-1 (mPGES-1) in human monocytes. Green and black tea extracts, and with a lower potency, Rooibos tea extract, inhibited lipopolysaccharide (LPS) and calcium ionophore-induced PGE2 formation. In addition, all tea extracts inhibited the LPS-induced expression of mPGES-1, and the green and black tea extracts also inhibited, to a lesser extent, COX-2 expression. The tea extracts only marginally reduced cPLA2 expression and had no effect on COX-1 expression. EGCG, present in green and black tea, and quercetin, present in all three teas, also inhibited PGE2 formation and expression of mPGES-1, COX-2 and cPLA2. Cell-based and cell-free assays were also performed to evaluate direct effects on the enzymatic activity of COX and PGE synthases. Mainly, the cell-free assay demonstrated partial inhibition by the tea extracts and polyphenols. However, the inhibition required higher doses compared to the effects demonstrated on protein expression. In conclusion, green and black tea, and to a lesser extent Rooibos tea, are potent inhibitors of PGE2 formation in human monocytes, and mediate their effects by inhibiting the expression of the enzymes responsible for PGE2 formation, especially mPGES-1.     

Bioassay-guided purification and identification of antimicrobial components in Chinese green tea extract

The Chinese green tea extract was found to strongly inhibit the growth of major food-borne pathogens, Escherichia coli O157:H7, Salmonella Typhimurium DT104, Listeria monocytogenes, Staphylococcus aureus, and a diarrhoea food-poisoning pathogen Bacillus cereus, by 44-100% with the highest activity found against S. aureus and lowest against E. coli O157:H7. A bioassay-guided fractionation technique was used for identifying the principal active component. A simple and efficient reversed-phase high-speed counter-current chromatography (HSCCC) method was developed for the separation and purification of four bioactive polyphenol compounds, epicatechin gallate (ECG), epigallocatechin gallate (EGCG), epicatechin (EC), and caffeine (CN). The structures of these polyphenols were confirmed with mass spectrometry. Among the four compounds, ECG and EGCG were the most active, particularly EGCG against S. aureus. EGCG had the lowest MIC90 values against S. aureus (MSSA) (58 mg/L) and its methicilin-resistant S. aureus (MRSA) (37 mg/L). Scanning electron microscopy (SEM) studies showed that these two compounds altered bacterial cell morphology, which might have resulted from disturbed cell division. This study demonstrated a direct link between the antimicrobial activity of tea and its specific polyphenolic compositions. The activity of tea polyphenols, particularly EGCG on antibiotics-resistant strains of S. aureus, suggests that these compounds are potential natural alternatives for the control of bovine mastitis and food poisoning caused by S. aureus.     

Redifferentiation of human hepatoma cells induced by green tea polyphenols

A novel approach for the treatment of cancer is the differentiation therapy in which cancer cells are induced to attain a mature phenotype when exposed to differentiation inducers. To examine the effects of polyphenols extracted from green tea, i.e. ( – )-epicatechin (EC), ( – )-epigallocatechin (EGC), ( – )-epicatechin gallate (ECG) and ( – )-epigallocatechin gallate (EGCG), on the proliferation and redifferentiation of human hepatoma cell line SMMC-7721, we measured the changes of cell growth, cell surface charge and cell morphography after treament with green tea polyphenols. It was found that the growth curve of treated cells was decreased remarkably, cell surface charge of treated cells was decreased and the microvilli on the surface of treated cells were reduced obviously. It confirmed that green tea polyphenols could reverse malignant phenotypic characteristics and induced redifferentiation of SMMC-7721 cells. The ability of green tea polyphenols to inhibit reactive oxygen species (ROS)-mediated oxidative damage of DNA was also assessedin vitro by measuring the conversion of supercoiled pBR322 plasmid DNA to the open circular and linear forms. It was found that green tea polyphenols could significantly inhibit the oxidative damage of DNA induced by a water-soluble azo initiator 2,2-azobis(2-amidinopropane hydrochloride) (AAPH). However, they could promote the oxidative damage of DNA induced by H₂O₂ and Fe²⁺ at high concentrations. The relationship between the anti-cancer activity and antioxidation-prooxidation activity of green tea polyphenols is discussed.     

Influence of Tea Brewing Parameters on the Antioxidant Potential of Infusions and Extracts Depending on the Degree of Processing of the Leaves of Camellia sinensis

The polyphenol content of tea depends on the growing region, harvest date, the production process used, and the brewing parameters. In this study, research was undertaken that included an analysis of the influence of the brewing process parameters on the content of total polyphenols (Folin-Ciocalteu), epigallocatechin gallate (HPLC), and antioxidant activity (against DPPH radicals) of fresh tea shrub leaves grown from Taiwan and of teas obtained from them (oolong, green in bags, and green loose from the spring and autumn harvest). The antioxidant potential was determined in the methanol and aqueous extracts, as well as in infusions that were obtained by using water at 65 or 100 °C and infusing the tea for 5 or 10 min. The highest content of total polyphenols and epigallocatechin gallate was found in green tea extracts from the spring harvest. However, in the case of infusions, the highest content of these compounds was found in green tea in bags. Steaming at 100 °C for 10 min, turned out to be the most favourable condition for the extraction. Oolong tea, brewed at 100 °C for 5 min was characterised by the highest antioxidant activity against stable DPPH radicals.     

Predictive models for tyrosinase inhibitors: Challenges from heterogeneous activity data determined by different experimental protocols

Quantitative Structure-Activity Relationship (QSAR) models of tyrosinase inhibitors were built using Random Forest (RF) algorithm and evaluated by the out-of-bag estimation (R²_OOB) and 10-fold cross validation (Q²_CV). We found that the performances of QSAR models were closely correlated with the systematic errors of inhibitory activities of tyrosinase inhibitors arising from the different measuring protocols. By defining ERR_sys, outliers with larger errors can be efficiently identified and removed from heterogeneous activity data. A reasonable QSAR model (R²_OOB of 0.74 and Q²_CV of 0.80) was obtained by the exclusion of 13 outliers with larger systematic errors. It is a clear example of the challenge for QSAR model that can overwhelm heterogeneous data from different experimental protocols.     

Biomimetic one-pot preparation of a black tea polyphenol theasinensin A from epigallocatechin gallate by treatment with copper(II) chloride and ascorbic acid

Chromatographic separation of black tea polyphenols is too difficult to supply sufficient quantities of pure compounds for biological experiments. Thus, facile methods to prepare black tea constituents were desired. Treatment of epigallocatechin gallate with copper(II) chloride efficiently afforded an unstable quinone dimer, dehydrotheasinensin A, and subsequent treatment with ascorbic acid stereoselectively yielded theasinensin A. The latter is a dimer with an R-biphenyl bond, one of the major polyphenols found in black tea. The method is simpler and more effective than enzymatic preparation.