原人参二醇纳米混悬剂的大鼠在体肠吸收研究

目的 考察原人参二醇纳米混悬剂的在体肠吸收特征。方法 采用溶剂蒸发法, 以白蛋白作为稳定剂制备原人参二醇纳米混悬剂。以酚红为标示物, 采用大鼠在体单向肠灌流法评价原人参二醇纳米混悬剂的大鼠在体肠吸收特性。结果 Zetasizer nano ZS仪测得原人参二醇纳米混悬剂平均粒径为(220±10)nm, Zeta电位为(-28±0.2)mV。肠吸收实验表明, 原人参二醇纳米混悬剂在整个肠段都有吸收, 且吸收速率常数(K_a)与渗透系数(P_eff)均大于原人参二醇原药(P<0.05), 原人参二醇纳米混悬剂在小肠段(十二指肠、空肠和回肠)的K_a与P_eff均大于结肠(P<0.05)。结论 原人参二醇纳米混悬剂能够有效促进原人参二醇的大鼠在体肠吸收, 其转运机制可能为主动转运或促进扩散。     

在体单向灌流法研究阿哌沙班肾衰大鼠肠吸收特性

目的 研究阿哌沙班在肾衰大鼠体内的肠吸收特性,并考察P糖蛋白(P-glycoprotein,P-gp)抑制剂对阿哌沙班吸收行为的影响。方法 选择肾衰大鼠在体单向灌流法进行肠吸收实验,建立大鼠阿哌沙班肠灌流液HPLC分析方法,以考察大鼠在体肠吸收影响因素。结果 阿哌沙班在各肠段的吸收速率常数（K_a）存在显著性差异（P<0.05）,但表观吸收系数（P_app）未见明显差异（P>0.05）;大鼠回肠段的K_a和P_app值随药物浓度的增加而降低;加入 P-gp抑制剂盐酸维拉帕米（0.1 mmol/L）后,阿哌沙班在空肠和回肠段的K_a和P_app值均明显增加。结论 阿哌沙班在各肠段均有吸收;P-gp抑制剂对阿哌沙班在空肠和回肠段的吸收均有明显的促进作用,表明阿哌沙班为P-gp底物,推测其吸收机制为主动转运。     

独活有效成分大鼠在体单向灌流肠吸收

为了考察独活提取物中二氢欧山芹醇乙酸酯、蛇床子素、二氢欧山芹醇当归酸酯等3种主要有效成分在大鼠的肠吸收性质,了解中药提纯后对大鼠肠吸收的影响，本文运用单向灌流模型并采用HPLC法测定独活提取物I(总香豆素含量＜10%)灌流液中3种成分在体肠灌流的浓度变化，并与独活提取物II(总香豆素含量≥60%)进行比较。结果表明二氢欧山芹醇乙酸酯、蛇床子素及二氢欧山芹醇当归酸酯的质量浓度分别为62～555 μg·mL^-1、101～887 μg·mL^-1和19～186 μg·mL^-1，吸收量与浓度呈线性关系,无高浓度饱和现象，吸收速率常数(K_a)、吸收渗透系数(P_app)值基本保持不变。3种成分在大鼠小肠主要以被动扩散方式吸收；在各肠段均有吸收，其中结肠吸收最好，各肠段的K_a、 P_app是结肠＞十二指肠＞空肠＞回肠，且结肠的K_a、 P_app值显著大于其他肠段；独活提取物II中3种成分的K_a、 P_app值显著小于独活提取物I中相应成分。     

水飞蓟宾在大鼠小肠中的吸收特性

目的考察水飞蓟宾在大鼠各肠段的吸收。方法以一定浓度的水飞蓟宾溶液作为灌流液，以0.1 mL·min^-1进行大鼠不同肠段的单向灌流,于不同时间收集肠灌流液并肝门静脉取血，分别用HPLC测定灌流液和血中药物浓度。结果实验结果表明190 μg·mL^-1水飞蓟宾在不同肠段的吸收速率常数(k_a)和有效透过系数(P_eff)是十二指肠>空肠>回肠>结肠。质量浓度为80 μg·mL^-1的水飞蓟宾灌流液在十二指肠的吸收与190和300 μg·mL^-1的吸收情况均有显著性差异(P<0.05)，但质量浓度为190 μg·mL^-1与300 μg·mL^-1的吸收之间无显著性差异(P>0.05)。肝门静脉血中药物分析也显示十二指肠>空肠>回肠>结肠。结论水飞蓟宾在小肠全肠道均有吸收且有高浓度饱和现象。     

基于平行人工膜渗透分析法对药物溶出/吸收仿生系统优化及其药物渗透性评价研究

目的 基于平行人工膜渗透分析法（Parallel artificial membrane permeability analysis,PAMPA）对药物溶出/吸收仿生系统（Drug dissolution and absorption simulating system,DDASS）的生物膜系统进行改进,以单硝酸异山梨酯、奥美拉唑和氢氯噻嗪为工具药进行渗透性考察,建立一种快速、简便的药物渗透性考察方法,以期进一步提高DDASS系统对药物渗透性评价的效率和准确性,并对药物体内吸收百分率进行预测。方法 以疏水性半透膜为载体,将1%大豆卵磷脂和0.1%胆固醇的正十二烷溶液涂于疏水性聚偏氟乙烯中空纤维膜上制备人工渗透膜,以人工渗透膜取代DDASS中的大鼠肠管,以BCS I类药单硝酸异山梨酯,BCS Ⅱ类药奥美拉唑和BCS Ⅲ类药氢氯噻嗪为工具药考察其在DDASS系统中经人工渗透膜的表观渗透系数（P_app）,采用大鼠在体单向肠灌流法对上述3种药物的有效渗透系数（P_eff）的测定,验证人工渗透膜对药物BCS归类的准确性。结果 单硝酸异山梨酯、奥美拉唑和氢氯噻嗪基于PAMPA优化DDASS法测得P_app分别为（3.644±0.291）×10^-6、（2.391±0.020）×10^-6、（0.129±0.032）×10^-6 cm/s;上述药物基于大鼠在体单向肠灌流法测得P_eff分别为（37.69±2.67）×10^-5、（33.72±5.02）×10^-5、（14.37±1.66）×10^-5 cm/s。三者的体内吸收百分率预测值分别为95.36%、65.76%、5.61%,与P_app呈正相关。结论 基于DDASS优化PAMPA法对3种不同BCS分类药物渗透性的考察结果与大鼠在体单向肠灌流结果一致,均证明单硝酸异山梨酯与奥美拉唑为高渗透性药物,氢氯噻嗪为低渗透性药物,符合FDA对三者的BCS分类。表明基于DDASS优化PAMPA法考察药物渗透性,结果准确,简单方便,节约资源,体内吸收百分率预测值具有参考价值,为仿制药申请生物等效性豁免及为创新药物口服生物利用度研究提供了可靠的体外预测平台。     

丙戊酸钠和丙戊酸镁在大鼠体内的药动学研究

目的 研究丙戊酸钠和丙戊酸镁在大鼠体内的药动学特征,评价其优势丙戊酸盐,为临床合理用药提供参考。方法 SD大鼠随机分为2组,分别灌胃给予丙戊酸钠片和丙戊酸镁片。于不同时间点眼眶取血。采用HPLC测定血清中丙戊酸的血药浓度,计算2种丙戊酸盐在大鼠体内的药动学参数,并比较2种丙戊酸盐之间的差异。结果 HPLC测定丙戊酸血药浓度方法专属性好,血清丙戊酸浓度在10.00~110.00 μg·mL^-1内线性关系良好。精密度、稳定性和回收率均符合要求。丙戊酸钠和丙戊酸镁在大鼠体内的主要药动学参数：T_1/2分别为（14.02±3.86） h和（12.11±1.95） h;T_max分别为（3.67±0.58） h和（2.67±0.26） h;C_max分别为（67.10±10.87）μg·mL^-1和（75.67±12.94）μg·mL^-1;AUC_（0-t）分别为（969.86±72.08）μg·mL^-1·h和（1 093.56±48.69）μg·mL^-1·h;AUC_（0-∞）分别为（1 178.10±185.29）μg·mL^-1·h和（1 279.35±109.18）μg·mL^-1·h;MRT_0-t分别为（10.73±2.05） h和（13.06±3.24） h。V_d分别为（16.31±2.18） L和（23.47±2.19） L;CL分别为（0.056 3±0.009） L·h^-1和（0.051 1±0.004） L·h^-1。结论 与丙戊酸钠相比,丙戊酸镁在大鼠体内的药动学参数具有一定的优势,可能是一种更具有治疗优势的丙戊酸盐。     

LC-MS/MS测定阿齐沙坦及其盐在大鼠血浆中的药动学研究

目的 建立液相色谱-串联质谱法测定大鼠血浆中阿齐沙坦及其盐的浓度并研究其药动学。方法 大鼠血浆样本以乙腈沉淀蛋白后,采用Eclipse Plus C₁₈色谱柱（50 mm×3.0 mm,1.8 μm）;流动相（乙腈：水=60：40）,流速为0.35 mL·min^-1,柱温为40℃;采用Agilent 6430三重四极杆串联质谱仪,离子化方式：电喷雾-正离子（API-ES）;监测方式：MRM;阿齐沙坦监测离子对457.3/233.1,缬沙坦监测离子对436.2/291.4,用作内标。SD大鼠灌胃给予阿齐沙坦1.0 mg·kg^-1及阿齐沙坦盐1.2 mg·kg^-1。结果 阿齐沙坦在5~30 000 ng·mL^-1内线性关系良好;回收率为85%~115%,精密度RSD在±15%内。阿齐沙坦盐大鼠体内主要动力学参数如下：AUC_{（0-24 h）}为（12.9±3.2）μg·mL^-1·h^-1,AUC_（0-∞）为（14.2±4.1）μg·mL^-1·h^-1,C_max为（3.8±0.3）μg·mL^-1,T_1/2为（13.4±0.5）h。阿齐沙坦的主要动力学参数如下：AUC_{（0-24 h）}为（8.1±2.6）μg·mL^-1·h^-1,AUC_（0-∞）为（9.7±3.1）μg·mL^-1·h^-1,C_max为（2.3±0.5）μg·mL^-1,T_1/2为（10.5±0.5）h。结论 本法经方法学验证,适用于大鼠血浆中阿齐沙坦及其盐的浓度测定,可用于阿齐沙坦及其盐大鼠体内药动学研究。     

青蒿琥酯自微乳在大鼠体内的药动学研究

目的 建立大鼠血浆中青蒿琥酯的HPLC-MS/MS测定方法,并研究青蒿琥酯自微乳在大鼠体内的药动学特征。方法 12只SD大鼠随机分为2组,单剂量分别灌胃（50 mg·kg^-1）青蒿琥酯自微乳和青蒿琥酯原料药,以格列吡嗪为内标,用LC-MS/MS测定给药后血浆中的药物浓度,并计算药动学参数。结果 青蒿琥酯血浆样品的线性范围为1.0~1 000.0 ng·mL^-1,回归方程为A=294.74C-439.33（r=0.999 6）,定量下限为1.0 ng·mL^-1。日内、日间变异系数（RSD）均<10%,符合生物样品的分析要求。青蒿琥酯原料药和青蒿琥酯自微乳的药动学参数C_max、t_1/2和AUC_0→t分别为：（87.6±8.80）ng·mL^-1,（1.88±0.33）h和（43.3±1.74）h·ng·mL^-1;（421±41.6）ng·mL^-1,（1.48±0.17）h和（282±17.7）h·ng·mL^-1。其中,C_max和AUC_0→t存在显著性差异（p<0.01）。结论 该方法简便灵敏,可用于血浆中青蒿琥酯的含量测定,经灌胃给药后,与原料药比较,青蒿琥酯自微乳能显著提高生物利用度。     

克拉霉素对Caco-2细胞转运他克莫司的影响及机制研究

目的 探讨克拉霉素对Caco-2细胞转运他克莫司的影响及机制。方法 用化学发光免疫分析法测定他克莫司药物浓度；建立Caco-2细胞单层模型，计算表观渗透系数，比较不同浓度他克莫司在Caco-2细胞单层中转运及不同浓度克拉霉素抑制他克莫司的转运。结果 他克莫司浓度为20，40和80 μg·mL^-1时吸收渗透系数P_appAP-BL分别为（2.47±0.09）×10^-6，（3.91±0.17）×10^-6和（4.49±0.16）×10^-6 cm·s^-1；分泌渗透系数P_appBL-AP分别为（6.05±0.21）×10^-6，（9.86±0.70）×10^-6和（11.75±0.28）×10^-6 cm·s^-1；外流比（apparent permeability ratio，PDR）分别为2.45±0.03，2.52±0.12和2.62±0.11；加入不同浓度的克拉霉素（15，30，60 μg·mL^-1）后，分泌渗透系数显著降低，而吸收系数影响不大，PDR随着克拉霉素浓度增加显著降低。结论 Caco-2细胞外排转运体可能参与了他克莫司的转运，克拉霉素合用他克莫司可显著影响他克莫司的吸收。     

细胞色素P450介导的氯沙坦钾对瑞格列奈在大鼠肝微粒体中代谢的影响

目的 研究瑞格列奈在大鼠肝微粒体中的酶促反应动力学，并考察氯沙坦钾对其在大鼠肝微粒体中代谢的影响。方法 建立大鼠肝微粒体体外孵育体系对瑞格列奈的代谢进行研究；以洛伐他汀为内标，应用UPLC测定大鼠肝微粒体中瑞格列奈的浓度。采用底物减少法，通过GraphPad Prism 5.0软件计算瑞格列奈的酶促反应动力学常数V_max和K_m；分别以系列浓度氯沙坦钾（2.5~50μmol·L^-1）与瑞格列奈（44 μmol·L^-1）于37℃水浴中共同孵育，并测定肝微粒体中瑞格列奈的减少量，考察氯沙坦钾对瑞格列奈的抑制作用。结果 瑞格列奈在大鼠肝微粒体的最佳孵育时间为40 min，最佳蛋白质量浓度为1 mg·mL^-1；瑞格列奈酶促反应动力学参数V_max=47.29μmol·min^-1·（mg·protein）^-1，K_m=51.41 μmol·L^-1；氯沙坦钾对瑞格列奈在体外肝微粒体抑制作用的IC₅₀值为17.89 μmol·L^-1。结论 氯沙坦钾对瑞格列奈在大鼠肝微粒体中的代谢具有较强的抑制作用，两药联合应用可能发生相互作用，具有诱发低血糖的风险。     

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.

---

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.