Integrating network pharmacology and pharmacological evaluation for deciphering the mechanism of (-)-epigallocatechin-3-gallate alleviating ethanol-inducedendothelial cells injury / 中国药理学与毒理学杂志

Objective To investigate the potential therapeutic targets and pharmacological mechanism of (-)-epigal?locatechin-3-gallate (EGCG) based on network pharmacology and experimental verification. METHODS The druggability of EGCG was measured by the traditional Chinese medicine systems pharmacology (TCMSP) server, and potential tar?gets of EGCG were identified by Pharm Mapper and Drug Repositioning and Adverse drug Reaction via Chemical-Pro?tein Interactome (DRAR-CPI). The potential targets were imported into GeneMANIA database to obtain the protein-pro?tein direct interaction network, and target physical interaction, co-expression, prediction, genetic interaction, and shared protein domains. The biological process, molecular functions, cellular components and KEGG signaling pathways of potential targets were analyzed using DAVID database. For further study, ethanol was used to establish a model of endothelial injury in vitro. The cell viability was assayed by MTT method, the cellular apoptosis was stained by Annexin V/PI, and the expression levels of Bcl-2, Bax and cleved-caspase-3 were tested by Western blotting. Then, JC-1 and nuclear translocation of NF-κB experiments were used to study the mitochondrial membrane potential and nuclear trans?location. RESULTS The oral availability of EGCG was 55.09％ (≥ 30％) and drug-like index was 0.77 (≥ 0.18), which were considered pharmacokinetically active. 17 potential targetable proteins of EGCG were predicted by Pharm Mapper and DRAR-CPI. Further research showed that 68.13％ displayed similar co-expression characteristics, 26.11％ physical interactions, and 2.74％ shared the same protein domain. The depth network analysis results showed that the biofunc?tions of EGCG were mainly by regulating glutathione derivative biosynthetic process, glutathione metabolic process, nitrogen compound metabolic process etc.. via drug binding, catalytic activity, glutathione transferase activity, anion bind?ing etc.. in sarcoplasmic reticulum, spindle pole, microtubule cytoskeleton and cytoplasm. KEGG enrichment analysis showed that Glutathione metabolism, IL-17 signaling pathway, EGFR tyrosine kinase inhibitor resistance, PI3K-Akt sig?naling pathway and other pathways were involves in the biofunction of EGCG. The above analyses indicated that EGCG exerts its biofunction through antioxidant and anti-inflammatory mechanisms. The experimental results showed that etha?nol 20.0 mmol·L-1 decreased cell viability, Bcl-2 expression, and increased cell apoptosis, the intracellular ROS, as well as the expression of Bax and cleaved-caspase-3 of human endothelial cells. However, treatment of the cells with EGCG can significantly alleviate ethanol induced endothelial cells injury. Further study showed that EGCG significantly allevi?ates ethanol induced mitochondrial depolarization and nuclear translocation of NF-κB. CONCLUSIONS EGCG exerts pharmacological efficacies on ethanol induced endothelial cell injury through multi-target, multi-function and multi-path?way mode. Protective effect of EGCG on ethanol induced cell injury was mainly through alteration of mitochondrial func?tion and NF-κB translocation. Therefore, EGCG have great potential in protecting against endothelial dysfunction of the persons who are chronically abuse of ethanol. This study also provides a new understanding of EGCG in clinical applica?tion on cardiovascular and cerebrovascular diseases.

Multiple target fishing hook prepared by photochemically coupling molecules in medicines onto magnetic nanoparticles / 中草药

Objective: To achieve multiple target fishing hook by efficiently grafting polymers containing benzophenone (BP) groups and photochemically coupling molecules in medicines onto magnetic nanoparticles (MNPs). Methods: MNPs attached carbonyl groups (Fe3O4-COOH) were prepared through a hydrothermal process. Then they were modified with DMSA, forming MNPs with thiol groups (Fe3O4-SH). Fe3O4-SH nanoparticles were grafted with polymer containing BP groups by surface-initiated condensation polymerization. Effects of monomer feed ratios and contents on the amounts of BP groups were investigated. The molecules in medicines were covalently coupled onto MNPs via photochemical reactions of BP groups. The contents of coupled molecules were determined by FT-IR and UV-Vis spectra analyses. Results: MNPs with average size of 100 nm were produced, modified with DMSA, and decorated by grafting polymer containing photosensitive BP groups. When the content of monomer containing BP groups was increased in grafting polymerizations, more BP groups were incorporated onto MNPs. This was conductive to the subsequent photo- coupling. FT-IR and UV-Vis spectra analyses confirmed the coupling of molecules in medicines. The active H and steric hinderance of the molecules affected their coupling. Conclusion: The resultant magnetic target fishing hook is ready as a probe for targets identification of traditional Chinese medicine (TCM).

Recent research advances of 1-deoxynojirimycin and its derivatives / 中国中药杂志

The 1-DNJ named 1-deoxynojirimycinis (2R,3R,4R,5S)-2-(hydroxymethyl) piperidine-3,4,5-triol, which is the nature active components existingin mulberryresources including leaves, stems, roots and silkworm larva, silkworm chrysalis, etc.The 1-deoxynojirimycin is a polyhydroxylated piperidine alkaloid, which was first found in Streptomyces as an antibiotic. Then the Japanese researchers isolated it from the mulberry root. 1-DNJ can inhibit postprandial hyperglycemia by suppressing intestinal alpha glucosidase. Therefore, 1-DNJ is often used to treat treating diabetes and complicating disease and to prevent obesity and weight-related disorders. With the development of the researches, 1-deoxynojirimycin and its derivtiv was discovered to possess anti-hyperglycemic, anti-virus, anti-tumor functions and so on. Therefore,based on our current studythe existing knowledge on source, technique preparation process, pharmacokinetics, bioactivties,and in silico target fishing of 1-DNJ were summarized, so that the researchers may use it to explore future perspective of research on 1-DNJ.

4-Phenyl-1, 3-thiazole-2-amines as scaffolds for new antileishmanial agents

There is still a need for new alternatives in pharmacological therapy for neglected diseases, as the drugs available show high toxicity and parenteral administration. That is the case for the treatment of leishmaniasis, particularly to the cutaneous clinical form of the disease. In this study, we present the synthesis and biological screening of eight 4-phenyl-1,3-thiazol-2-amines assayed against Leishmania amazonensis. Herein we propose that these compounds are good starting points for the search of new antileishmanial drugs by demonstrating some of the structural aspects which could interfere with the observed activity, as well as suggesting potential macromolecular targets. Methods: The compounds were easily synthesized by the methodology of Hantzsch and Weber, had their purities determined by Gas Chromatography-Mass spectrometry and assayed against the promastigote forms of Leishmania amazonensis as well as against two white cell lines (L929 and THP-1) and the monkey's kidney Vero cells. PrestoBlue® and MTT viability assays were the methodologies applied to measure the antileishmanial and cytotoxic activities, respectively. A molecular modeling target fishing study was performed aiming to propose potential macromolecular targets which could explain the observed biological behavior. Results: Four out of the eight compounds tested exhibited important anti-promastigote activity associated with good selectivity indexes when considering Vero cells. For the most promising compound, compound 6, IC50 against promastigotes was 20.78 while SI was 5.69. Compounds 3 (IC50: 46.63 µM; SI: 26.11) and 4 (IC50: 53.12 µM; SI: 4.80) also presented important biological behavior. A target fishing study suggested that S-methyl-5-thioadenosine phosphorylase is a potential target to these compounds, which could be explored to enhance activity and decrease the potential toxic side effects. Conclusions: This study shows that 4-phenyl-1,3-thiazol-2-amines could be good scaffolds to the development of new antileishmanial agents. The S-methyl-5-thioadenosine phosphorylase could be one of the macromolecular targets involved in the action.(AU)

In silico target fishing for the potential bioactive components contained in Huanglian Jiedu Tang (HLJDD) and elucidating molecular mechanisms for the treatment of sepsis / 中国天然药物

The present study was designed to target fish for potential bioactive components contained in a Huang Lian Jie Du decoction (HLJDD) and identify the underlying mechanisms of action for the treatment of sepsis at the molecular level. he bioactive components database of HLJDD was constructed and the sepsis-associated targets were comprehensively investigated. The 3D structures of the PAFR and TXA2R proteins were established using the homology modelling (HM) method, and the molecular effects for sepsis treatment were analysed by comparing the bioactive components database and the sepsis targets using computational biology methods. The results of the screening were validated with biological testing against the human oral epidermal carcinoma cell line KB in vitro. We found that multiple bioactive compounds contained in the HLJDD interacted with multiple targets. We also predicted the promising compound leads for sepsis treatment, and the first 28 compounds were characterized. Several compounds, such as berberine, berberrubine and epiberberine, dose-dependently inhibited PGE2 production in human KB cells, and the effects were similar in the presence or absence of TPA. This study demonstrates a novel approach to identifying natural chemical compounds as new leads for the treatment of sepsis.