Systematic screening of hepatoprotective components from traditional Chinese medicine: Zuojin Pill as an example.

ETHNOPHARMACOLOGICAL RELEVANCE: Zuojin Pill (ZJP), composed of Coptis chinensis Franch. and Euodia ruticarpa (A. Juss.) Benth. in a mass ratio of 6:1, is a famous traditional Chinese medicine (TCM) formula recorded in "Danxi's Experiential Therapy", an ancient medical book from the Ming Dynasty of China. It is used to treat liver fire invading the stomach, which is caused by liver stagnation transforming into fire and disharmony between the liver and stomach. AIM OF THE STUDY: To develop a systematic strategy to screen hepatoprotective components from TCM using ZJP as a model sample. MATERIALS AND METHODS: A CCl4-induced mouse model of acute liver injury was used for the verification of the hepatoprotective effects of ZJP. UPLC-Q-Exactive Plus Orbitrap MS/MS was used for the identification of the components in mouse serum after intragastric administration of ZJP. The hepatoprotective activities of the components found in mouse serum were tested in primary cultured mouse hepatocytes induced by CCl4. RESULTS: Nine components with significant hepatoprotective activity including berberine, epiberberine, coptisine, palmatine, jatrorrhizine, rutaecarpin, dehydroevodiamine, evocarpine and chlorogenic acid were successfully screened out. CONCLUSIONS: Our developed strategy has the advantages of high efficiency and low cost, and would provide a powerful tool for screening potential hepatoprotective components from TCM.

Study on quality control of Zuojin pill by HPLC fingerprint with quantitative analysis of multi-components by single marker method and antioxidant activity analysis.

Current quality control methods for Zuojin Pill (ZJP) lack comprehensiveness and practicability. This study aimed to develop a comprehensive strategy for the quality evaluation of ZJP and the prediction of potential bioactive components in ZJP. First, an HPLC method with excellent separation of main components was developed and was used to establish the chromatographic fingerprint of ZJP. Similarities were calculated by comparing 28 batches of ZJPs with the reference fingerprint and the resulting similarity values were all greater than 0.976. The 28 samples were classified into different groups according to their origins by Hierarchical Cluster Analysis, Principal component analysis, and orthogonal partial least squares discriminant analysis. Based on the classification, eight quality markers (Q-Markers) affecting the quality of ZJP were discovered. Then, using berberine as an internal standard substance, quantitative analysis of multi-components by single marker method (QAMS) for the determination of eight Q-markers was developed. The results showed that there was no significant difference between QAMS and external standard method (P＞0.05). Finally, using an off-line antioxidant system and partial least-squares model (PLS), the fingerprint-efficacy relationship of ZJP was constructed to explore and predict the bioactive components in ZJP. The present study strategy could be also applied to comprehensive quality study of other TCMs.

Intermolecular C-H Aminocyanation of Indoles via Copper-iodine Cocatalyzed Tandem C-N/C-C Bond Formation.

An efficient copper-iodine cocatalyzed intermolecular C-H aminocyanation of indoles with a broad substrate scope has been developed for the first time. This method enables highly step-economic access to 2-amino-3-cyanoindoles in moderate to good yields and provides a complementary strategy for the regioselective difunctionalization of carbon═carbon double bonds of interest in organic synthesis and related areas. Mechanistic studies suggest that these transformations are initiated by iodine-mediated C2-H amination with azoles, followed by copper-catalyzed C3-H cyanation with ethyl cyanoformate.

Fast Screening of Biomembrane-Permeable Compounds in Herbal Medicines Using Bubble-Generating Magnetic Liposomes Coupled with LC-MS.

A novel strategy based on the use of bionic membrane camouflaged magnetic particles and LC-MS was developed to quickly screen the biomembrane-permeable compounds in herbal medicines. The bionic membrane was constructed by bubble-generating magnetic liposomes loaded with NH4HCO3 (BMLs). The lipid bilayer structure of the liposomes enabled BMLs to capture biomembrane-permeable compounds from a herbal extract. The BMLs carrying the compounds were then separated from the extract by a magnetic field. Upon heat treatment, NH4HCO3 rapidly decomposed to form CO2 bubbles within the liposomal bilayer, and the captured compounds were released from BMLs and analyzed by LC-MS. Jinlingzi San (JLZS), which contains various natural ingredients, was chosen to assess the feasibility of the proposed method. As a result, nine potential permeable compounds captured by BMLs were identified for the first time. Moreover, an in vivo animal study found that most of the compounds screened out by the proposed method were absorbed into the blood. The study provides a powerful tool for rapid and simultaneous prediction of multiple biomembrane-permeable components.

Rapid determination of seven bioactive components in rat plasma by UPLC-MS/MS and its application to pharmacokinetic compatibility study of Jinlingzi San.

Jinlingzi San (JLZS), composed of Fructus Toosendan (FT) and Rhizoma Corydalis (RC), is a classical traditional Chinese medicine prescription for regulating Qi to relieve pain. The present study investigated the pharmacokinetic compatibility of FT and RC in JLZS. A fast, selective and sensitive UPLC-MS/MS method for simultaneous determination of one limonoid (toosendanin), four tertiary alkaloids (corydaline, tetrahydropalmatine, tetrahydrocoptisine, tetrahydroberberine) and two quaternary alkaloids (palmatine, dehydrocorydaline) in rat plasma was established and fully validated. The plasma samples were pretreated by a fast protein precipitation and chromatographed using a 1.7-µm C18 column and 0.1 % formic acid-water and acetonitrile via gradient elution with a run time of 3.7 min. Multiple reaction monitoring mode with positive electrospray ionization was adopted to detect the analytes and internal standard (diphenhydramine). The lower limits of quantification were 0.08-3.09 ng/mL using only 50 µL of plasma sample. Using the proposed method, the pharmacokinetic differences of seven bioactive components in rats after administration of JLZS and the single herb (FT or RC) were investigated. The results showed that the elimination of toosendanin and alkaloids decreased significantly in the JLZS group (p < 0.05) compared with the single herb group, and the exposure of the alkaloids increased in some degree. The study demonstrated the synergistic effect of combining FT with RC on the pharmacokinetics of seven bioactive components and provided new information for a better understanding of the compatibility mechanism of JLZS.

Pharmacokinetic Studies of Three Alkaloids in Rats After Intragastrical Administration of Lycopodii Herba Extract by LC-MS/MS.

Lycopodii Herba is a widely used traditional medicinal herb, and contains diverse fascinating alkaloids. In this study, a fast and sensitive LC-MS/MS method for the simultaneous determination of lycodoline, α-obscurine, and N-demethyl-α-obscurine from Lycopodii Herba in rat plasma and brain tissue was developed and validated. Biological samples were extracted via a protein precipitation procedure using methanol as the extraction solvent and Huperzine B as the internal standard. Chromatographic separation was carried out using a Thermo Syncronis-C18 column (50 mm × 2.1 mm, 5 µm) and a gradient mobile phase containing methanol and water with 0.05% formic acid. The three alkaloids were detected by positive electrospray ionization in selective reaction monitoring mode. The selectivity, crosstalk, carryover effect, linearity, accuracy, precision, extraction recovery, matrix effect, and stability of the current method were validated. Then, using the validated method, the plasma pharmacokinetics and brain tissue distribution of the alkaloids in rats were investigated after intragastrical administration of Lycopodii Herba extract. The three alkaloids were shown to be rapidly absorbed into the blood (Tmax, 0.79-1.58 h), and then also eliminated rapidly (t1/2, 1.27-2.24 h). All of them could pass through the blood-brain barrier. The method provides a new research approach to expand preclinical studies of Lycopodii Herba.

A new agent developed by biotransformation of polyphyllin VII inhibits chemoresistance in breast cancer.

Biotransformation by the endophytes of certain plants changes various compounds, and this 'green' chemistry becomes increasingly important for finding new products with pharmacological activity. In this study, polyphyllin VII (PPL7) was biotransformed by endophytes from the medicinal plant Paris polyphylla Smith, var. yunnanensis. This produced a new compound, ZH-2, with pharmacological activity in vitro and in vivo. ZH-2 was more potent than PPL7 in selectively killing more chemoresistant than chemosensitive breast cancer cells. ZH-2 also re-sensitized chemoresistant breast cancer cells, as evidenced by the improved anti-cancer activity of commonly-used chemotherapeutic agent in vitro, in vivo, and in clinical samples. This anti-chemoresistance effect of ZH-2 was associated with inhibiting the epithelial-mesenchymal transition (EMT) pathway. Taken together, our findings are the first one to link biotransformation with a biomedicine. The results provide insights into developing new pharmacologically-active agents via biotransformation by endophytes.