基于UPLC-QTOF/MS的茵陈蒿汤体外成分鉴定和急性肝损伤防护作用入血成分研究

鉴定茵陈蒿汤体外活性成分和体内入血成分,为探究其对急性肝损伤防护作用和防治机制提供了科学依据。利用超高效液相色谱-四极杆飞行时间质谱(UPLC-QTOF/MS)技术,鉴定茵陈蒿汤水煎液及口服预防给药14天后,刀豆蛋白A (con-A)所致急性肝损伤小鼠的血清样本(本研究中所有实验动物的使用获得海军军医大学伦理委员会批准,批号19YF1459400)中的化学成分。初步鉴定茵陈蒿汤水煎液含90个化合物,主要为黄酮类、萜类、鞣质类、醌类等,血清样本中含5个原形成分,包括大黄酚、脱氧土大黄苷-8-O-没食子酸盐、玉叶金花苷酸、7-甲氧基-香豆素、大黄素。本文所建立的UPLC-QTOF/MS方法能高效灵敏地鉴别茵陈蒿汤的体外成分及其入血成分,初步阐明茵陈蒿汤保肝作用的药效物质基础,可为茵陈蒿汤的质量标志物筛选和药理作用研究提供科学依据。     

反相高效液相色谱法测定大鼠血浆中6,7-二甲氧基香豆素含量及其在药动学中的应用

目的 :建立大鼠血浆中 6 ,7 二甲氧基香豆素反相高效液相色谱紫外检测方法 ,并研究其在大鼠体内的药动学过程。方法 :血浆样品经乙腈沉淀蛋白 ,上清液在 5 0℃下经氮气流吹干 ,残渣加入流动相 10 0 μL溶解后 ,离心 ,直接进样。流动相为甲醇 1%乙酸水溶液 四氢呋喃 (30∶6 3∶7)。检测波长为 340nm。大鼠经灌胃予 6 ,7 二甲氧基香豆素后 ,所得血药浓度数据用Topfit程序拟合并估算药动学参数。结果 :线性范围为 0 0 2 5～ 2 5 μg·mL-1。经非房室模型计算 ,6 ,7 二甲氧基香豆素主要药动学参数T1/ 2 为 0 7h ,AUC0→t为 16 5 0 6ng·h·mL-1,AUC0→∞ 为 172 8 9ng·h·mL-1,CLtot为 5 4 0L·h-1,用Topfit程序拟合 ,其体内过程符合二室开放模型。结论 :本法操作简便 ,可应用于大鼠血浆中 6 ,7 二甲氧基香豆素浓度的测定及其药动学研究。     

基于方剂配伍含药血清“谱-效关系”的茵陈蒿汤保肝作用药效物质研究

目的：初步阐明茵陈蒿汤保肝作用的药效物质。方法：SD大鼠灌胃制备茵陈蒿汤及其各单味药、各缺味方含药血清,一方面采用已建立的方法测定不同方剂配伍含药血清HPLC指纹图谱;另一方面建立H₂O₂诱导的HL-7702肝细胞损伤模型,并对不同方剂配伍含药血清进行保肝作用药效评价,最后分别采用双变量相关性分析和多元回归分析,指认含药血清指纹图谱中与药效相关的色谱峰。结果：茵陈蒿汤含药血清指纹图谱中6号峰与肝细胞保护作用明显相关,1~6,8,9,11,20号峰与肝细胞保护作用有一定关系。结论：茵陈蒿汤保肝作用的药效物质主要包括来源于大黄的蒽醌类成分及体内代谢产物、来源于栀子的环烯醚萜类成分及体内代谢产物和西红花酸类成分体内代谢产物,其中1个蒽醌类成分体内代谢产物活性较强。     

枸橼果实的香豆素和黄酮类成分研究

目的研究枸橼果实的化学成分。方法用硅胶柱色谱法及Sephadex LH-20柱色谱法进行分离纯化,依据理化性质和光谱数据鉴定化合物结构。结果从枸橼果实的95%乙醇提取物中分离鉴定了11个化合物,包括7个香豆素类化合物:7-羟基香豆素(1)、5,7-二羟基香豆素(2)、7-羟基-6-甲氧基香豆素(3)、5,7-二甲氧基香豆素(4)、6,7-二甲氧基香豆素(5)、佛手柑内酯(6)和8-(2',3'-二羟基-3'-甲丁基)-5,7-二甲氧基香豆素(7);4个黄酮类化合物:香叶木素(8)、柚皮素(9)、柚皮苷(10)和橙皮素(11)。结论化合物1,2,5～9为首次从枸橼中分离得到。     

中医药信息

中医药信息茵陈五苓散的临床和生物利用度研究茵陈五苓散出自《金匮要略》，由茵陈蒿末１０克，五苓散５克所组成，其有效成分主要是６、７一二甲氧基香豆素。茵陈五苓散主要功效有清热，利尿和退黄。日本研究证明，此药对癌细胞有抑制作用，与化疗药合用可提高对某些肿瘤...     

1-［1-(6-甲氧基-2-萘基)乙基］-2-(4-硝基苄基)-6,7-二甲氧基-1,2,3,4-四氢异喹啉氢溴酸盐的光降解产物研究

研究1-［1-(6-甲氧基-2-萘基)乙基］-2-(4-硝基苄基)-6,7-二甲氧基-1,2,3,4-四氢异喹啉氢溴酸盐(编号P91024)遇光后颜色变暗的光降解产物。采用HPLC-MS及波谱分析鉴定光降解产物的化学结构，并经有机反应合成对照验证。P91024光降解的3个主要产物分别为溴化N-(4-硝基苄基)-6,7-二甲氧基-3,4-二氢异喹啉、1-［1-(6-甲氧基-2-萘基)乙基］-6,7-二甲氧基-1,2,3,4-四氢异喹啉和2-异丙基-6-甲氧基萘。     

赛灵斯

[通用名称] donepezil hydrochloride,盐酸多奈哌齐 ［化学名称］　1-苄基-4-［（5，6-二甲氧基-1-茚满酮-2-基）甲基］哌啶盐酸盐.     

蒙药铁杆蒿的定性和定量分析

目的：建立铁杆蒿的TLC鉴别及HPLC定量分析方法。方法：采用TLC和HPLC法对采集于内蒙古通辽市5个不同地区铁杆蒿进行定性和定量分析。TLC条件：对照品为8-羟基-6,7-二甲氧基香豆素和7-羟基-6-甲氧基香豆素,吸附剂为硅胶G,展开剂为二氯甲烷-丙酮(7:1),显色剂为紫外灯下观察;对照品为6,8-二甲氧基香豆素-7-O-β-D-葡萄糖苷,吸附剂为硅胶G,展开剂为二氯甲烷-甲醇（5:1）,显色剂为紫外灯下观察。 HPLC条件：色谱柱为Topsil C18 (250 mm × 4.6 mm, 5 μm);流动相由水（A）和乙腈（B）组成。检测波长为263 nm,柱温为30 ℃。结果：对照品8-羟基-6,7-二甲氧基香豆素、7-羟基-6-甲氧基香豆素和6,8-二甲氧基香豆素-7-O-β-D-葡萄糖苷与不同样品具有Rf值相同的特征斑点,重复性好,易于鉴别;在选用的HPLC条件下,所检测化合物（1-7）表现出良好的线性关系（r>0.999 3）,其平均回收率分别为98.5、95.0、97.0、98.3、96.0、98.9、99.1,RSD均小于2.0%。结论：建立了一种简便、稳定和可靠的铁杆蒿定性定量分析方法,为综合评价该药材质量提供依据。     

芫花根中新双香豆素的分离与鉴定

目的研究芫花根(Daphne genkwa)的化学成分。方法采用硅胶和Sephadex LH-20柱色谱技术进行分离纯化，运用多种波谱分析进行结构鉴定。结果分离得到一个新的双香豆素异西瑞香素，结构被确认为7-羟基-6-甲氧基-4［(2-氧杂-2H-1-苯并吡喃-7-基)-氧基］-2H-1-苯并吡喃-2-酮。结论异西瑞香素(isodaphnoretin)为一个新化合物。     

基于含药血清经时谱效关系的茵陈蒿汤保肝作用药效物质研究

目的：阐明茵陈蒿汤的保肝作用药效物质。方法：以SD大鼠为供体制备茵陈蒿汤灌胃给药后10个时间点含药血清,高效液相色谱法（HPLC）测定含药血清指纹图谱,采用HL-7702肝细胞损伤模型对含药血清进行保肝作用药效评价,分别采用药时曲线和时效曲线比较及双变量相关性分析的谱-效关系分析方法指认含药血清指纹图谱中与药效相关的色谱峰。结果：药时曲线和时效曲线比较发现,含药血清指纹图谱中4~7、20号峰对损伤肝细胞具有明显保护作用,8、11~19号峰对损伤肝细胞具有保护作用,但可能也具有一定肝细胞毒性有关,1~3及9号峰具有肝细胞毒性;双变量相关性分析发现,含药血清指纹图谱中1~9、15~17、20号峰对损伤肝细胞具有一定保护作用,其中4、5、20号峰活性较强,6号峰活性最强。结论：茵陈蒿汤保肝作用的药效物质主要包括来源于大黄的蒽醌类成分、来源于栀子的环烯醚萜类成分和西红花酸类成分及以上三类成分的体内代谢产物,其中1个蒽醌类成分、1个蒽醌类成分代谢产物和1个西红花酸类成分代谢产物活性较强,另1个蒽醌类成分代谢产物活性最强。但以上三类成分剂量较高时也具有一定肝细胞毒性,即具有肝细胞保护和毒性双重作用。     

Pharmacological treatment of COVID-19: an opinion paper

The precocity and efficacy of the vaccines developed so far against COVID-19 has been the most significant and saving advance against the pandemic. The development of vaccines has not prevented, during the whole period of the pandemic, the constant search for therapeutic medicines, both among existing drugs with different indications and in the development of new drugs. The Scientific Committee of the COVID-19 of the Illustrious College of Physicians of Madrid wanted to offer an early, simplified and critical approach to these new drugs, to new developments in immunotherapy and to what has been learned from the immune response modulators already known and which have proven effective against the virus, in order to help understand the current situation.     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

Lung disease and PKCs

The lung offers a rich opportunity for development of therapeutic strategies focused on isozymes of protein kinase C (PKCs). PKCs are important in many cellular responses in the lung, and existing therapies for pulmonary disorders are inadequate. The lung poses unique challenges as it interfaces with air and blood, contains a pulmonary and systemic circulation, and consists of many cell types. Key structures are bronchial and pulmonary vessels, branching airways, and distal air sacs defined by alveolar walls containing capillaries and interstitial space. The cellular composition of each vessel, airway, and alveolar wall is heterogeneous. Injurious environmental stimuli signal through PKCs and cause a variety of disorders. Edema formation and pulmonary hypertension (PHTN) result from derangements in endothelial, smooth muscle (SM), and/or adventitial fibroblast cell phenotype. Asthma, chronic obstructive pulmonary disease (COPD), and lung cancer are characterized by distinctive pathological changes in airway epithelial, SM, and mucous-generating cells. Acute and chronic pneumonitis and fibrosis occur in the alveolar space and interstitium with type 2 pneumocytes and interstitial fibroblasts/myofibroblasts playing a prominent role. At each site, inflammatory, immune, and vascular progenitor cells contribute to the injury and repair process. Many strategies have been used to investigate PKCs in lung injury. Isolated organ preparations and whole animal studies are powerful approaches especially when genetically engineered mice are used. More analysis of PKC isozymes in normal and diseased human lung tissue and cells is needed to complement this work. Since opposing or counter-regulatory effects of selected PKCs in the same cell or tissue have been found, it may be desirable to target more than one PKC isozyme and potentially in different directions. Because multiple signaling pathways contribute to the key cellular responses important in lung biology, therapeutic strategies targeting PKCs may be more effective if combined with inhibitors of other pathways for additive or synergistic effect. Mechanisms that regulate PKC activity, including phosphorylation and interaction with isozyme-specific binding proteins, are also potential therapeutic targets. Key isotypes of PKC involved in lung pathophysiology are summarized and current and evolving therapeutic approaches to target them are identified.     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.