α-乳香酸通过抑制TLR4介导的炎症通路改善急性肾损伤

α-乳香酸(α-BA)是乳香药材经过提取、分离、纯化后的产物,可用于治疗多种炎症性疾病。本研究旨在通过体内外实验评估α-BA在改善缺血再灌注损伤诱导的急性肾损伤中的作用。对大鼠进行右肾切除和左肾缺血再灌注损伤,损伤前0.5 h给予α-BA处理。体内实验,对组织进行肾功能、组织病理学和TUNEL染色分析。体外实验,对近端肾小管细胞(HK-2)进行氧糖剥夺(OGD)操作,以模仿缺血再灌注损伤,并检测细胞存活率、免疫荧光染色及Toll样受体4(TLR4)和核因子(NF)-κB表达。结果显示,给予α-BA的模型组大鼠血清肌酐和血尿素氮水平显著降低,并缓解细胞凋亡和肾脏形态学改变;经α-BA处理的HK-2细胞,NF-κBp6的表达明显降低。表明α-BA改善缺血再灌注损伤诱导的急性肾损伤,部分原因是通过TLR4介导的炎症通路促进肾小管细胞存活,从而改善肾功能。

Antioxidant properties of magnesium lithospermate B contribute to the cardioprotection against myocardial ischemia/reperfusion injury in vivo and in vitro

OBJECTIVE: To determine the cardioprotective effect of magnesium lithospermate B (MLB) on myocardial ischemia/reperfusion (MI/R) injury and to investigate the antioxidant potential in vivo and in vitro. METHODS: MI/R injury was induced by the occlusion of left anterior descending coronary artery for 30 min followed by reperfusion for 3 h in rats. After reperfusion, hearts were harvested to assess infarct size, histopathological damages, the levels of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), reduced glutathione (GSH) and malondialdehyde (MDA). Blood samples were col- lected to determine serum levels of creatine kinase-MB (CK-MB), cardiac troponin (cTnI) and lactate dehydrogenase (LDH). Furthermore, simulatedischemia/reperfusion (SI/R) injury in vitro was established by oxygen and glucose deprivation (OGD) for 2 h followed by 24-hour recovery period in cardiomyocytes. The activity of LDH in the cultured supernatant and the levels of intracellular reactive oxygen species (ROS), SOD and MDA in cardiomyocytes were also measured. Finally, cardiomyocytes apoptosis was determined with flow cytometry. RESULTS: MLB significantly limited infarct size, ameliorated histopathological damages and prevented leakage of CK-MB, cTnI and LDH. Additionally, SOD, CAT, GPx and GSH activities were notably increased by MLB, along with the MDA content decreased as compared with the model group in rats. In vitro study, MLB also decreased LDH activity in the cultured supernatant, increased SOD activity in cardiomyocytes, reduced intracellular ROS and MDA levels, and significantly suppressed cardiomyocytes apoptosis. CONCLUSION: MLB possessed remarkably cardioprotective effects on MI/R injury in vivo and in vitro. The protection of MLB may contribute to its antioxidant properties.

Simultaneous determination of ivabradine and N-desmethylivabradine in human plasma and urine using a LC-MS/MS method: application to a pharmacokinetic study

A sensitive and specific liquid-chromatography tandem mass spectrometry(LC-MS/MS)assay has been developed and validated for the simultaneous quantification of ivabradine and its active metabolite N-desmethylivabradine in human plasma and urine.The assay employed a single liquid–liquid extraction of the analytes from plasma and urine samples,and diazepam was used as internal standard(IS).The chromatographic separation was achieved on a Diamonsil C18 column(150 mm4.6 mm,5 mm,Dikma)using a mixture of methanol and aqueous 5 mM ammonium acetate buffer containing 0.2%formic acid(80:20,v/v)as mobile phase.The assay for ivabradine and N-desmethylivabradine in plasma showed good linearity(r≥0.99)over the ranges 0.1013–101.3 ng/mL and 0.085–25.5 ng/mL,respectively.The assay for ivabradine and N-desmethylivabradine in urine showed good linearity(r≥0.99)over the ranges 10.13–6078 ng/mL and 8.5–850 ng/mL,respectively.The intra-and inter-day accuracy and precision values were found to be within the assay variability limits(RSD<15%)in accordance with FDA guidelines.The methods were successfully used for evaluating the pharmacokinetic properties of ivabradine and N-desmethylivabradine in human plasma and urine in Chinese healthy volunteers.

Determination of ifenprodil by LC–MS/MS and its application to a pharmacokinetic study in healthy Chinese volunteers

This paper reports the development and validation of an assay for ifenprodil based on liquid chromatography–tandem mass spectrometry(LC–MS/MS)and its application to a pharmacokinetic study involving single and multiple intravenous infusions to healthy Chinese volunteers.After sample preparation of plasma by liquid–liquid extraction with ethyl acetate,the analyte and internal standard,urapidil,were separated by reversed phase chromatography in a run time of 4 min and detected by positive ion electrospray ionization followed by multiple reaction monitoring of the precursor-to-product ion transitions at m/z 326.2-308.1 for ifenprodil and m/z 388.4-205.3 for IS.The assay was linear in the concentration range 0.2–50.0 ng/mL with recovery 476.4%.In the pharmacokinetic study of single intravenous infusions of 5,10 and 15 mg ifenprodil,peak plasma concentrations and areas under the plasma concentration–time curve were both linearly related to dose.In the pharmacokinetic study of multiple once daily intravenous infusions of 10 mg ifenprodil for 7 days,pharmacokinetic parameters were similar to those after the single dose showing that ifenprodil does not accumulate on repeated administration.

Safety and efficacy of meplazumab in healthy volunteers and COVID-19 patients: a randomized phase 1 and an exploratory phase 2 trial

Recent evidence suggests that CD147 serves as a novel receptor for severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)infection.Blocking CD147 via anti-CD147 antibody could suppress the in vitro SARS-CoV-2 replication.Meplazumab is a humanized anti-CD147 IgG_(2) monoclonal antibody,which may effectively prevent SARS-CoV-2 infection in coronavirus disease 2019(COVID-19)patients.Here,we conducted a randomized,double-blinded,placebo-controlled phase 1 trial to evaluate the safety,tolerability,and pharmacokinetics of meplazumab in healthy subjects,and an open-labeled,concurrent controlled add-on exploratory phase 2 study to determine the efficacy in COVID-19 patients.In phase 1 study,59 subjects were enrolled and assigned to eight cohorts,and no serious treatment-emergent adverse event(TEAE)or TEAE grade≥3 was observed.The serum and peripheral blood Cmax and area under the curve showed non-linear pharmacokinetic characteristics.No obvious relation between the incidence or titer of positive anti-drug antibody and dosage was observed in each cohort.The biodistribution study indicated that meplazumab reached lung tissue and maintained>14 days stable with the lung tissue/cardiac blood-pool ratio ranging from 0.41 to 0.32.In the exploratory phase 2 study,17 COVID-19 patients were enrolled,and 11 hospitalized patients were involved as concurrent control.The meplazumab treatment significantly improved the discharged(P=0.005)and case severity(P=0.021),and reduced the time to virus negative(P=0.045)in comparison to the control group.These results show a sound safety and tolerance of meplazumab in healthy volunteers and suggest that meplazumab could accelerate the recovery of patients from COVID-19 pneumonia with a favorable safety profile.