小檗碱的酶反应动力学及其代谢酶表型和代谢产物研究

目的 采用混合人肝微粒体和重组人源细胞色素酶P450（CYP）同工酶,研究小檗碱的代谢特性,明确参与小檗碱代谢的CYP酶亚型及其贡献度,并确证相关代谢产物的结构。方法 将混合人肝微粒体分别与20、100、200、400、600、800、1 200 ng/mL的小檗碱共同孵育后,计算小檗碱酶反应动力学米氏常数（K_m）、最大反应速率（V_max）、体外肝微粒体对药物的固有清除率（CL_int）;采用CYP酶的特异性抑制剂研究小檗碱的代谢表型;将重组人源CYP同工酶CYP3A4、CYP1A2、CYP2D6、CYP2C9与一定质量浓度的小檗碱孵育,UPLC法测定孵育液中原形药物的剩余量和代谢产物的生成量;整体归一化法计算各酶的代谢贡献度;LC-MS/MS法鉴定相关代谢产物。结果 小檗碱在混合人肝微粒体中的V_max为1.51 nmol?mg^?1?h^?1;K_m为2.69 nmol/mL;CL_int为0.56 mL?mg^?1?h^?1。CYP2D6的特异性抑制剂奎尼丁和CYP1A2的特异性抑制剂呋喃茶碱对小檗碱的代谢有显著抑制作用,其他CYP酶抑制剂对小檗碱的代谢无显著影响。CYP2D6、CYP1A2对小檗碱代谢产物M₁（去亚甲基小檗碱）的贡献度分别为75.253 9%、23.323 6%;对M₂产物（唐松草分定或小檗红碱）的贡献度分别为46.893 8%、8.679 5%。小檗碱在体外混合人肝微粒体温孵体系中的主要代谢途径为O-去甲基化,可生成去亚甲基小檗碱和唐松草分定或小檗红碱。结论 小檗碱在肝脏中主要被CYP2D6和CYP1A2代谢,分别生成去亚甲基小檗碱、唐松草分定或小檗红碱。     

In Vitro Characterization of Glucuronidation of Vanillin: Identification of Human UDP‐Glucuronosyltransferases and Species Differences

Vanillin is a food flavoring agent widely utilized in foods, beverages, drugs, and perfumes and has been demonstrated to exhibit multiple pharmacological activities. Given the importance of glucuronidation in the metabolism of vanillin, the UDP‐glucuronosyltransferase conjugation pathway of vanillin was investigated in this study. Vanillin glucuronide was identified by high‐performance liquid chromatography‐tandem mass spectrometry (HPLC‐MS/MS) and a hydrolysis reaction catalyzed by β‐glucuronidase. The kinetic study showed that vanillin glucuronidation by HLMs and HIMs followed Michaelis‐Menten kinetics and the kinetic parameters were as follows: 134.9 ± 13.5 μM and 81.3 ± 11.3 μM for K_m of HLMs and HIMs, 63.8 ± 2.0 nmol/min/mg pro and 13.4 ±2.0 nmol/min/mg pro for V_max of HLMs and HIMs. All UDP‐glucuronosyltransferase (UGT) isoforms except UGT1A4, 1A9, and 2B7 showed the capability to glucuronidate vanillin, and UGT1A6 exerted the higher V_max/K_m values than other UGT isoforms for the glucuronidation of vanillin when assuming expression of isoforms is similar in recombinant UGTs. Kinetic analysis using liver microsomes from six studied speices indicated that vanillin had highest affinity for the monkey liver microsomes enzyme (K_m = 25.6 ± 3.2 μM) and the lowest affinity for the mice liver microsomes enzyme (K_m = 149.1 ± 18.4 μM), and intrinsic clearance was in the following order: monkey > dog > minipig > mice > rat ~ human. These data collectively provided important information for understanding glucuronidation of vanillin. Copyright © 2012 John Wiley & Sons, Ltd.     

黄独素B的体外代谢通路及其代谢产物研究

目的基于体外代谢模型对黄独素B的代谢稳定性、主要CYP450代谢酶表型及其代谢产物进行研究。方法黄独素B分别在人肝微粒体(HLM)和大鼠肝微粒体(RLM)中共同孵育,采用UPLC-MS/MS检测孵育液中剩余的黄独素B含量,分析其在HLM和RLM中的代谢稳定性。利用10种重组人CYP450酶(1A1、1A2、1B1、2A13、2A6、2B6、2D6、2C9、2C19、3A4)分别与黄独素B共同孵育,确定其代谢酶表型,结合大鼠离体肝灌流模型对黄独素B的主要代谢酶表型进行确认。此外,分别对HLM和RLM孵育体系中的黄独素B代谢产物进行定性分析,考察黄独素B在HLM和RLM中代谢产物的区别。结果在HLM和RLM中,黄独素B的转化率分别为37%、59%,经HLM和RLM代谢的体外半衰期(t1/2)为97.4、52.3 min,推算得到的HLM和RLM中的固有清除率(CLint, in vivo)为8.23、23.9 mL/(min·kg),肝清除率(CLh)为5.89、16.8 mL/(min·kg),由此可知黄独素B在RLM体系中的代谢转化速率较HLM中快。黄独素B体外代谢酶表型结果可知其I相代谢是由多个CYP同工酶介导的,包括3A4、2C19、2C9、1A13及1A1,其中CYP3A4对黄独素B的代谢起主导作用;肝灌流实验结果显示,随着酮康唑给药剂量的增加,对肝脏中CYP3A4的抑制作用增强,黄独素B在肝脏中的代谢减少,在灌流液中的剩余量增加,印证了CYP3A4对黄独素B的代谢作用。此外,2种肝微粒体孵育后的黄独素B都只产生了1个代谢产物(M1),为黄独素B去甲基化产物。结论黄独素B在RLM中的代谢转化速率较HLM中快。黄独素B的主要代谢酶表型为CYP3A4,其在HLM和RLM中产生的代谢物均为去甲基化产物。     

UPLC-MS/MS研究披针灰叶素B在不同种属肝微粒体中的代谢稳定性及代谢酶表型

应用体外肝微粒体孵育体系,研究披针灰叶素B(lanceolatin B)在大鼠、人、比格犬、猴肝微粒体中的代谢稳定性及体外代谢参数,确定主要代谢披针灰叶素B的CYP酶表型。将披针灰叶素B与不同种属肝微粒体共同孵育,应用UPLC-MS/MS检测孵育液中披针灰叶素B的含量,考察其代谢稳定性与体外代谢动力学参数。将披针灰叶素B与各CYP450同工酶CYP2E1,2C19,1A2,2D6,2C9,3A4,2A1的特异性抑制剂共同孵育,确定其代谢酶表型。研究结果显示,披针灰叶素B在人、比格犬肝微粒体中不代谢,而在大鼠与猴肝微粒体中可代谢,它们的体外半衰期(T_(1/2))与固有清除率(CL_(int))分别为11.57,8.07min和0.12,0.17 m L·min-1·mg-1,结果没有显著性差异;披针灰叶素B在肝微粒体中的代谢存在种属差异,在大鼠肝微粒体中由多个酶共同参与代谢,其中CYP1A2酶的作用最强,是主要代谢披针灰叶素B的酶。     

不同化学环境下的马钱子碱和士的宁在大鼠肝微粒体内代谢动力学研究

目的研究马钱子碱(BRU)与士的宁(STR)单体、马钱子总生物碱和马钱子提取物中活性成分BRU与STR在大鼠肝微粒体中的酶促反应动力学,考察BRU与STR单体、马钱子总生物碱和提取物中的BRU与STR在大鼠肝微粒体中的代谢差异。方法采用LC-MS/MS法测定BRU和STR单体、总生物碱和提取物种BRU和STR在体外代谢系统中的含量变化,并计算其酶促反应动力学参数。结果与BRU与STR单体组相比,马钱子总生物碱和提取物中2种单体成分的最大反应速度(Vmax)和米氏常数(Km)值均显著降低;与马钱子总碱组相比,马钱子提取物中2种单体成分的Vmax和Km值均显著升高。马钱子总生物碱组、单体成分组和马钱子提取物组中BRU的清除率(CLint)之间无明显差异;而三者中STR的CLint依次减少。结论马钱子总碱、马钱子提取物中的BRU和STR及BRU与STR单体均可被肝微粒体代谢,且各组中BRU和STR的代谢动力学参数具有明显差异。     

5种黄连生物碱大鼠体外肝代谢特征比较

目的 探讨黄连中5种结构相似异喹啉生物碱大鼠体外肝代谢的选择性。方法 采用大鼠体外肝微粒体温孵方法,通过考察温孵时间、微粒体蛋白浓度以及底物浓度对小檗碱、巴马汀、黄连碱、表小檗碱和药根碱代谢的影响,求得代谢反应动力学参数K_m、V_max和CL_int。结果 5种黄连生物碱成分的体外肝代谢动力学参数K_m和CL_int间差异明显（P＜0.05）。结论 黄连中5种异喹啉生物碱大鼠体外肝代谢存在显著选择性。     

Strong Specific Inhibition of UDP‐glucuronosyltransferase 2B7 by Atractylenolide I and III

Drug‐metabolizing enzymes inhibition‐based drug–drug interaction remains to be the key limiting factor for the research and development of efficient herbal components to become clinical drugs. The present study aims to determine the inhibition of uridine 5′‐diphospho‐glucuronosyltransferases (UGTs) isoforms by two important efficient herbal ingredients isolated from Atractylodes macrocephala Koidz, atractylenolide I and III. In vitro recombinant UGTs‐catalysed glucuronidation of 4‐methylumbelliferone was used to determine the inhibition capability and kinetics of atractylenolide I and III towards UGT2B7, and in silico docking method was employed to explain the possible mechanism. Atractylenolide I and III exhibited specific inhibition towards UGT2B7, with negligible influence towards other UGT isoforms. Atractylenolide I exerted stronger inhibition potential than atractylenolide III towards UGT2B7, which is attributed to the different hydrogen bonds and hydrophobic interactions. Inhibition kinetic analysis was performed for the inhibition of atractylenolide I towards UGT2B7. Inhibition kinetic determination showed that atractylenolide I competitively inhibited UGT2B7, and inhibition kinetic parameter (Ki) was calculated to be 6.4 μM. In combination of the maximum plasma concentration of atractylenolide I after oral administration of 50 mg/kg atractylenolide I, the area under the plasma concentration‐time curve ration AUC_i/AUC was calculated to be 1.17, indicating the highly possible drug–drug interaction between atractylenolide I and drugs mainly undergoing UGT2B7‐catalysed metabolism. Copyright © 2015 John Wiley & Sons, Ltd.     

In vitro cytotoxic and toxicological activities of ethanolic extract of Kaempferia galanga Linn. and its active component,ethyl-p-methoxycinnamate,against cholangiocarcinoma

ObjectiveTo evaluate the cytotoxic, apoptotic, mutagenic and immunomodulatory activities of Kaempferia galanga Linn. (KG) extract and ethyl-p-methoxycinnamate (EPMC) in vitro.MethodsThe present study investigated the cytotoxic [using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide test], apoptotic (using a mitochondrial membrane potential assay), mutagenic (using a micronucleus test) and immunomodulatory (using flow cytometry) activities of the ethanolic extract of KG and its bioactive component, EPMC, against two cholangiocarcinoma (CCA) cell lines, CL-6 and HuCCT1, and one normal human cell line, OUMS-36T-1F.ResultsBoth KG extract and EPMC exhibited moderate cytotoxic activity against both CCA cells. The cytotoxic activity was supported by their concentration-dependent induction of apoptosis. CL-6 was most sensitive (3–4 fold) and selective to 5-fluorouracil (5-FU), compared with KG extract and EPMC [median half inhibiting concentration (IC₅₀) and selectivity index (SI) were 23.01 μg/mL and 17.32; 78.41 μg/mL and 4.44; 100.76 μg/mL and 2.20, respectively for 5-FU vs. KG extract vs. EPMC]. HuCCT1 was relatively more sensitive and selective to 5-FU and EPMC than KG extract [median IC₅₀ and SI were 66.03 μg/mL and 6.04; 60.90 μg/mL and 3.65; 156.60 μg/mL and 2.23, respectively for 5-FU vs. EPMC vs. KG extract]. EPMC produced relatively potent cytotoxic activity against polymorphonuclear cells (IC₅₀ = 92.20 μg/mL). KG extract and EPMC exhibited concentration-dependent mutagenic activity, as well as inhibition of tumor necrosis factor-α and interleukin-6.ConclusionConsidering cytotoxic, apoptotic, immunomodulatory and mutagenic activities, further development of KG as a drug candidate is likely to focus on the oral pharmaceutical formulation of a standardized KG extract rather than isolated compounds.     

采用肝微粒体研究夫拉平度葡萄糖醛酸化代谢的性别及种属差异

 目的 研究夫拉平度（flavopiridol）在不同种属不同性别肝微粒体中葡萄糖醛酸化（UDP-glucuronosyl transferase,UGT）代谢速率及代谢机制。方法 采用人和4种动物（小鼠、大鼠、豚鼠、狗）雌雄两种性别的肝微粒体酶对夫拉平度的酶代动力学进行研究,以超高效液相色谱法测定葡萄糖醛酸化代谢反应物的浓度,比较葡萄糖醛酸化代谢速率。结果 夫拉平度在各种肝微粒体中都能被代谢并生成葡萄糖醛酸结合物。同一性别的5种动物的肝微粒体代谢夫拉平度的速率有种属差异（P     

人尿苷二磷酸葡糖醛酸转移酶(UGT)亚型参与反式-白藜芦醇体外代谢的研究

目的:研究参与反式-白藜芦醇(trans-resveratrol,TR)Ⅱ相代谢的主要尿苷二磷酸葡糖醛酸转移酶(UGTs)亚型。方法:在体外对反式-白藜芦醇与12种主要的人重组UGT亚型进行温孵,采用液相色谱-质谱法测定其葡萄糖醛酸代谢产物,对其结构做初步分析,并考察不同UGT亚型对白藜芦醇代谢产物生成速率的影响。结果:在体外代谢系统中,白藜芦醇被UGT催化生成2种单葡萄糖醛酸代谢产物M-1和M-2,初步推断其为白藜芦醇-4’和3-葡萄糖醛酸化物,亚型UGT1A1,1A3,1A8,1A9,1A10都参与了催化产生代谢产物M-1和M-2,UGT1A6,1A7仅对M-2的生成有贡献。随着底物浓度的升高,UGT1A1,1A10催化底物产生M-1和M-2及1A8催化底物产生M-2的速率都减慢,出现了底物抑制现象。结论:UGT1A1,1A8,1A9,1A10参与了代谢产物M-1的产生,其中UGT1A9的贡献最大,UGT1A1,1A6,1A7,1A8,1A9,1A10参与了代谢产物M-2的产生,其中1A1和1A9贡献最大,UGT1A3也有少量参与2种代谢物的产生,其他亚型几乎都不参与反式-白藜芦醇的Ⅱ相代谢反应。     

效应面法优化蒙花苷水解工艺及其水解物的醛糖还原酶抑制活性研究

目的：用效应面法对蒙花苷的水解工艺进行优化,通过醛糖还原酶模型研究其水解物金合欢素醛糖还原酶的抑制活性。方法：建立水解物金合欢素体外生物酶模型,通过测定NADPH的荧光吸收,计算水解物金合欢素对醛糖还原酶的抑制率,从而得到水解物的醛糖还原酶抑制活性的IC₅₀的平均值。在蒙花苷水解工艺中采用效应面法优化水解工艺,并对结果进行预测分析。结果：水解物金合欢素醛糖还原酶抑制活性,其IC₅₀的平均值为2.74 mg·L^-1。优选的水解工艺为水解时间7.4 h,硫酸甲醇0.54 mol·L^-1,固液比3：1。结论：水解物金合欢素具有比蒙花苷较好的醛糖还原酶抑制活性。优选的蒙花苷水解工艺简便,预测性良好。     

基于UGT1A1酶介导的胆红素代谢考察大黄素在肝微粒体体系中的肝毒性

以胆红素代谢过程中UGT1A1酶介导的胆红素葡萄糖醛酸结合环节为切入点,通过考察待测物大黄素对该酶的抑制作用预测其肝毒性。以胆红素为UGT1A1酶底物,于人肝微粒体、大鼠肝微粒体及人重组UGT1A1酶中加入不同浓度胆红素及大黄素,分别以胆红素的总代谢产物生成量对胆红素底物浓度作图,以米氏方程双倒数法绘图,并以不同曲线的斜率对应胆红素底物浓度绘制slop图计算表观抑制常数Ki,考察其对胆红素葡萄糖醛酸结合的抑制作用,预测其肝毒性有无及大小。结果显示大黄素在3个体系中对UGT1A1酶均有中强抑制作用,且抑制类型均为竞争型抑制。HLM,RLM,r UGT1A1体系的Ki分别为(5.400±0.956),(10.020±0.611),(4.850±0.528),P均0.05。同时发现,大黄素对于UGT1A1酶的抑制在大鼠及人之间无明显种属差异。大黄素可通过抑制UGT1A1酶活性导致胆红素代谢异常,从而存在引发肝毒性的潜在危险。该试验所建立的体外研究方法为中药肝毒性药物的筛选提供了新思路和新方法,对中药安全性评价具有借鉴意义。     

Comparison of Inhibition Capability of Scutellarein and Scutellarin Towards Important Liver UDP‐Glucuronosyltransferase (UGT) Isoforms

Scutellarin is an important bioactive flavonoid extracted from Erigeron breviscapus (Vant.) Hand‐Mazz, and scutellarein is the corresponding aglycone of scutellarin. The present study aims to compare the inhibition potential of scutellarin and scutellarein towards several important UDP‐glucuronosyltransferase (UGT) isoforms, including UGT1A1, UGT1A6, UGT1A9 and UGT2B7. It was demonstrated that scutellarein exerted stronger inhibition towards the tested UGT isoforms than scutellarin. Furthermore, the inhibition kinetic type and parameters (K_i) were determined for the scutellarein's inhibition towards these UGT isoforms. Competitive inhibition of scutellarein towards all these UGT isoforms was demonstrated, and the K_i values were calculated to be 0.02, 5.0, 5.8 and 35.9 μM for UGT1A1, 1A6, 1A9 and 2B7, respectively. Using in vivo maximum plasma concentration of scutellarein in rat, the in vitro–in vivo extrapolation was performed to predict in vivo situation, indicating the most possible in vivo adverse effects due to the inhibition of scutellarein towards UGT1A1. All these results remind us to monitor the utilization of scutellarin and scutellarein, and the herbs containing these two components. Copyright © 2013 John Wiley & Sons, Ltd.     

The Inhibitory Effect of 20(S)‐Protopanaxatriol (ppt) Towards UGT1A1 and UGT2B7

Ginseng, a commonly used natural product, has been frequently reported to induce herb–drug interaction with many clinical drugs. The intestinal bacterial metabolites of ginsenosides have been widely regarded as the substance basis for ginseng–drug interactions. To date, little is known about the inhibitory effect of intestinal bacterial metabolites of ginsenosides towards UDP‐glucuronosyltransferases (UGTs). In vitro investigation of the inhibition of 20(S)‐protopanaxatriol (ppt) towards UGT1A1 and UGT2B7 was carried out. The results showed that ppt exhibited strong noncompetitive inhibition towards UGT1A1 and competitive inhibition towards UGT2B7. The inhibition kinetic parameters (K_i) were calculated to be 8.8 and 2.2 μM for UGT1A1 and UGT2B7, respectively. Using the maximum plasma concentration of ppt, the alteration of area under the concentration–time curve was calculated to be 20% and 70% respectively for UGT1A1‐mediated and UGT2B7‐mediated metabolism. However, given that the varied contribution of these two UGT isoforms towards drug metabolism and the influence of herb complexity and individual difference, the explanation of these results should be paid more caution. Copyright © 2012 John Wiley & Sons, Ltd.     

Investigation of UDP-glucuronosyltransferases (UGTs) inhibitory properties of carvacrol

UDP‐glucuronosyltransferases (UGTs), the most important phase II drug metabolizing enzymes (DMEs), could metabolize many drugs and various endogenous substances including bilirubin, steroid hormones, thyroid hormones, bile acids and fat‐soluble vitamins. Evaluation of the inhibitory effects of compounds on UGTs is clinically important because inhibition of UGT isoforms could not only result in serious drug–drug interactions (DDIs), but also induce metabolic disorders of endogenous substances. The aim of the present study was to investigate the inhibitory effects of carvacrol on major UGT isoforms. The results showed that carvacrol could inhibit the activity of UGT1A9 with negligible effects on other UGT isoforms. When 4‐methylumbelliferone (4‐MU) was used as a nonspecific probe substrate and recombinant UGT enzymes were utilized as an enzyme resource, the inhibition of UGT1A9 was best fit to the competitive type and the inhibition kinetic parameter (K_i) was calculated to be 5.7 µ m . Furthermore, another specific probe substrate, propofol, was employed to determine the inhibitory kinetics of UGT1A9, and the results demonstrated that the inhibitory type was noncompetitive. The inhibition kinetic parameter (K_i) was determined to be 25.0 µ m . Because this substrate‐dependent inhibition of UGT1A9 might confuse the in vitro–in vivo extrapolation, these in vitro inhibition kinetic parameters should be interpreted with special caution. Copyright © 2011 John Wiley & Sons, Ltd.     

Combination of a biopharmaceutic classification system and physiologically based pharmacokinetic models to predict absorption properties of baicalein in vitro and in vivo

ObjectiveTo determine the in vitro and in vivo absorption properties of active ingredients of the Chinese medicine, baicalein, to enrich mechanistic understanding of oral drug absorption.MethodsThe Biopharmaceutic Classification System (BCS) category was determined using equilibrium solubility, intrinsic dissolution rate, and intestinal permeability to evaluate intestinal absorption mechanisms of baicalein in rats in vitro. Physiologically based pharmacokinetic (PBPK) model commercial software GastroPlus™ was used to predict oral absorption of baicalein in vivo.ResultsBased on equilibrium solubility, intrinsic dissolution rate, and permeability values of main absorptive segments in the duodenum, jejunum, and ileum, baicalein was classified as a drug with low solubility and high permeability. Intestinal perfusion with venous sampling (IPVS) revealed that baicalein was extensively metabolized in the body, which corresponded to the low bioavailability predicted by the PBPK model. Further, the PBPK model predicted the key indicators of BCS, leading to reclassification as BCS-II. Predicted values of peak plasma concentration of the drug (C_max) and area under the curve (AUC) fell within two times of the error of the measured results, highlighting the superior prediction of absorption of baicalein in rats, beagles, and humans. The PBPK model supported in vitro and in vivo evidence and provided excellent prediction for this BCS class II drug.ConclusionBCS and PBPK are complementary methods that enable comprehensive research of BCS parameters, intestinal absorption rate, metabolism, prediction of human absorption fraction and bioavailability, simulation of PK, and drug absorption in various intestinal segments across species. This combined approach may facilitate a more comprehensive and accurate analysis of the absorption characteristics of active ingredients of Chinese medicine from in vitro and in vivo perspectives.     

LC-MS/MS研究新型酪氨酸酶抑制剂UP302在大鼠肝微粒体中代谢的酶动力学

 目的 建立并验证测定大鼠肝微粒体孵育液中UP302含量的高效液相色谱-串联质谱法,研究UP302在大鼠肝微粒体中代谢的酶动力学。方法 经大鼠肝微粒体孵育的UP302样品,经色谱柱分离,电喷雾离子化负离子检测,扫描方式为选择反应监测,用于定量分析的离子反应分别为m/z 301.1→135.2（UP302）和m/z 252.9→132.0（内标大豆苷元）。考察不同孵育时间、底物浓度和微粒体浓度对UP302代谢的影响,确定最佳反应条件。结果 标准曲线线性范围为0.1～20 μmol·L-1,线性关系良好,日内日间准确度在86.42%～112.59%,日内日间精密度均小于9.09%,回收率在100.50%～109.91%之间。确定了酶动力学参数,最大反应速度V_max为196.08 μmol·L-1·min-1·g-1 ,米氏常数K_m为14.98 μmol·L-1,内在代谢清除率CL_int为13.09 mL-1·min-1·g-1。结论 该检测方法快速、专属、灵敏度高,可满足酶动力学检测要求。     

Potential medicinal plants involved in inhibiting 3CLpro activity: A practical alternate approach to combating COVID-19

At present, a variety of vaccines have been approved, and existing antiviral drugs are being tested to find an effective treatment for coronavirus disease 2019 (COVID-19). However, no standardized treatment has yet been approved by the World Health Organization. The virally encoded chymotrypsin-like protease (3CL^pro) from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which facilitates the replication of SARS-CoV in the host cells, is one potential pharmacological target for the development of anti-SARS drugs. Online search engines, such as Web of Science, Google Scholar, Scopus and PubMed, were used to retrieve data on the traditional uses of medicinal plants and their inhibitory effects against the SARS-CoV 3CL^pro. Various pure compounds, including polyphenols, terpenoids, chalcones, alkaloids, biflavonoids, flavanones, anthraquinones and glycosides, have shown potent inhibition of SARS-CoV-2 3CL^pro activity with 50% inhibitory concentration (IC₅₀) values ranging from 2–44 µg/mL. Interestingly, most of these active compounds, including xanthoangelol E (isolated from Angelica keiskei), dieckol 1 (isolated from Ecklonia cava), amentoflavone (isolated from Torreya nucifera), celastrol, pristimerin, tingenone and iguesterin (isolated from Tripterygium regelii), tannic acid (isolated from Camellia sinensis), and theaflavin-3,3′-digallate, 3-isotheaflav1in-3 gallate and dihydrotanshinone I (isolated from Salvia miltiorrhiza), had IC₅₀ values of less than 15 µg/mL. Kinetic mechanistic studies of several active compounds revealed that their mode of inhibition was dose-dependent and competitive, with K_i values ranging from 2.4–43.8 μmol/L. Given the significance of plant-based compounds and the many promising results obtained, there is still need to explore the phytochemical and mechanistic potentials of plants and their products. These medicinal plants could serve as an effective inexpensive nutraceutical for the general public to help manage COVID-19.     

清开灵注射液及其主要成分黄芩苷和栀子苷对人肝微粒体CYP450酶6种亚型的体外抑制作用研究

 目的 研究清开灵注射液及其主要成分黄芩苷和栀子苷对人肝微粒体CYP1A2、CYP2C8、CYP2C9、CYP2C19、CYP2D6和CYP3A4的体外抑制作用。方法 采用液相色谱-串联质谱法（LC-MS/MS）同时测定对乙酰氨基酚、6α-羟基紫杉醇、4-羟基双氯芬酸、4-羟基美芬妥英、右啡烷、1-羟基咪达唑仑和6β-羟基睾酮,分别代表CYP1A2、CYP2C8、CYP2C9、CYP2C19、CYP2D6和CYP3A4的活性;清开灵注射液、黄芩苷、栀子苷和7种混合探针底物在人肝微粒体中共同孵育,并计算其IC₅₀值表示对CYP450酶的抑制程度。结果 在人肝微粒体体外孵育体系中,清开灵注射液对CYP1A2的IC₅₀值为0.6%,对其他亚型的IC₅₀值从1.1%到6.0%;黄芩苷和栀子苷对CYP450酶6种亚型的IC₅₀值均大于100 μmol·L-1。结论 在正常剂量下,清开灵注射液对人肝微粒体CYP1A2可能有抑制作用,对人肝微粒体CYP2C8、CYP2C9、CYP2C19、CYP2D6和CYP3A4无明显抑制作用;黄芩苷和栀子苷对CYP450酶6种亚型均无抑制作用。