目的 研究将淫羊藿产地初加工与饮片炮制相结合的一体化(以下简称"一体化")加工技术,优选最佳的一体化工艺.方法 对采收期的淫羊藿鲜药材分别采用3种加工炮制方法:①淫羊藿鲜药材在烘箱50℃下烘至不同含水量(0、10、20、30、40、50、60、70、80、90、100％)时,分别取出切丝,加20％的羊油炮制得11个新型饮片(S1-S11);②淫羊藿鲜药材晒干后,取干药材,闷润,切丝,自然晒干,加20％的羊油炮制得传统饮片(S12);③淫羊藿鲜药材在不同温度(30、40、50、60、70、80、90、100、110、120℃)分别烘至含水量为30％(七成干)时,分别取出切丝,加20％的羊油得10个新型饮片(S13-S22).采用HPLC法测定各样品中的5种主要黄酮朝藿定A、朝藿定B、朝藿定C、淫羊藿苷、宝藿苷I(以下简称"5种黄酮")的含量,比色法测定总黄酮的含量,并同时测定各饮片稀乙醇提取物的自由基清除活性.结果 饮片S20的5种黄酮和总黄酮含量均最高,且S20清除DPPH自由基和超氧阴离子自由基的能力最强.综合考虑,优选的一体化工艺为100℃烘至含水量为七成干时,切丝,加20％的羊油进行炮制.结论 优选的一体化加工方法较传统方法省时、省力,新型饮片的活性成分含量较高、自由基清除能力较强.

The Keap1 (Kelch-like ECH-associated protein 1)-Nrf2 (nuclear factor erythroid 2-related factor 2)-ARE (antioxidant response element) pathway is the major defending mechanism against oxidative stresses, and directly disrupting the Keap1-Nrf2 protein-protein interaction (PPI) has been an attractive strategy to target oxidative stress-related diseases, including cardiovascular diseases. Here, we describe the design, synthesis, and structure-activity relationships (SARs) of indoline-based compounds as potent Keap1-Nrf2 PPI inhibitors. Comprehensive SAR analysis and thermodynamics-guided optimization identified 19a as the most potent inhibitor in this series, with an IC50 of 22 nM in a competitive fluorescence polarization assay. Further evaluation indicated the proper drug-like properties of 19a. Compound 19a dose-dependently upregulated genes and protein level of Nrf2 as well as its downstream markers and showed protective effects against lipopolysaccharide-induced injury in both H9c2 cardiac cells and mouse models. Collectively, we reported here a novel indoline-based Keap1-Nrf2 PPI inhibitor as a potential cardioprotective agent.

Introduction: The transcription factor Nrf2 (nuclear factor erythroid 2-related factor 2) is the first line of defense against a plethora of environmental or endogenous deviations in redox metabolism, proteostasis, inflammation, etc. Therefore, pharmacological activation of Nrf2 is a potential therapeutic approach for several diseases related to oxidative stress and inflammation, such as cancer, cardiovascular, and neurodegenerative diseases.Areas covered: The authors first describe the biological function of Nrf2 and the molecular regulatory mechanism of Keap1-Nrf2-ARE ((Kelch-like ECH-Associating protein 1)-Nrf2-(antioxidant response element)). Then, they review recent progress of covalent activators and non-covalent Keap1-Nrf2 protein-protein interaction (PPI) inhibitors from patents and publications in 2017-present, consisting of new chemical molecules, structure optimization of reported activators and progress in preclinical or clinical trials.Expert opinion: Despite significant achievements in the development of Nrf2 activators, the selectivity is the primary consideration. Due to reacting with redox-sensitive cysteines in proteins except for Keap1, electrophilic activators often exhibit off-target effects. For Keap1-Nrf2 PPI inhibitors, how to enhance in vivo efficacy and/or penetrate blood-brain barrier (BBB) to reach central nervous system (CNS) is also challenging. Fragment-based drug discovery (FBDD), carboxylic acid bioisosteric replacement and prodrug approach might be used to circumvent this challenge. Moreover, the possibility of cancer risk caused by Nrf2 activation needs to be considered carefully.

Reversibly altering endogenous protein levels are persistent issues. Herein, we designed photoswitchable azobenzene-proteolysis targeting chimeras (Azo-PROTACs) by including azobenzene moieties between ligands for the E3 ligase and the protein of interest. Azo-PROTACs are light-controlled small-molecule tools for protein knockdown in cells. The light-induced configuration change can switch the active state to induce protein degradation activity, which can be reversely controlled by light exposure in intact cells. We compared the protein degradation abilities of Azo-PROTACs with different configurations and linker lengths. Using the stable form with the best degradation ability against the BCR-ABL fusion and ABL proteins in myelogenous leukemia K562 cells, we showed that Azo-PROTAC combines the potent protein knockdown and facile cell uptake properties of the small-molecule PROTAC with a reversible photoswitchability, offering a promising chemical knockdown strategy based on the light-induced reversible on/off properties.

Elevated plasma total homocysteine (tHcy) level has been acknowledged as an independent risk factor for cardiovascular diseases. Salvianolic acid A (SalA) is one of the major active water-soluble ingredients of Salvia miltiorrhiza. The pharmacokinetics (PK) and pharmacodynamics (PD) of SalA on plasma tHcy in methionine-loading rats was evaluated. Acute treatment with a single dose of SalA (1, 2.5, 5 mg/kg) in rats with Met loading significantly reduced the plasma tHcy level in a dose-dependent manner. In order to characterize the effect of SalA on plasma tHcy, a mechanism-based PK-PD model was developed. Modeling and simulations well fitted the measured values. By comparison with the PK-PD model of danshensu (DSS), it was found that the elevated effect of SalA on tHcy by methylation was weaker than that of DSS, while trans-sulfuration promotion effect of SalA on tHcy was greater than that of DSS.

To establish population pharmacokinetics model of nebivolol given by oral route in Chinese healthy volunteers using nonlinear mixed effect model (NONMEM), thus evaluating the factors that could influence clinical pharmacokinetics of nebivolol. Clinical data from 25 healthy volunteers were collected and the experiments included single-dose and multiple-dose administration. Volunteers were given 5 mg nebivolol orally and plasma concentrations of the drug were determined by LC-MS/MS. The population pharmacokinetic modeling was performed with NONMEM program and estimated by bootstrap procedure. The results showed that two-compartment model had fit for nebivolol pharmacokinetics. Inter-individual variability was described by multiplicative model. Typical values of central volume of distribution and clearance were 1.84 × 10 3 L and 7.68 × 10 2 L/h. The population pharmacokinetic parameters were mainly influenced by body weight, diastolic blood pressure, blood urea nitrogen and phenotype. The parameters estimated from nonparametric bootstrap procedure were comparable to those from NONMEM. The established population pharmacokinetics model could explain the reasons for the plasma concentration of individual variations of nebivolol, which could be used to guide clinical administration to Chinese people.