Flavonoids are key bioactive components for evaluating the pharmacological activities of Chimonanthus praecox. Exploring the potential flavonoids and pharmacological mechanisms of C. praecox lays a foundation for the rational development and efficient utilization of this plant. This study employed ultra-performance liquid chromatography-tandem mass spectrometry-based widely targeted metabolomics to comprehensively identify the flavonoids in C. praecox. Network pharmacology was employed to explore the bioactive flavonoids and their mechanisms of action. Molecular docking was adopted to validate the predicted results. Finally, the content of bioactive flavonoids in different varieties of C. praecox was measured. The widely targeted metabolomics analysis identified 387 flavonoids in C. praecox, and the flavonoids varied among different varieties. Network pharmacology predicted 96 chemical components including 19 bioactive compounds, 181 corresponding targets and 2 504 disease targets, among which 99 targets were shared by the active components and the disease. Thirty-three core targets were predicted, involving 229 gene ontology terms and 99 pathways (P≤0.05), which indicated that the flavonoids components of C. praecox exhibited pharmacological activities including antioxidant, anti-inflammatory, antimicrobial, and antiviral activities. Topological analysis screened out five core components (salvigenin, laricitrin, isorhamnetin, quercetin, and 6-hydroxyluteolin) and five core targets (SRC, PIK3R1, AKT1, ESR1, and AKR1C3). The predicted bioactive flavonoids from C. praecox stably bound to key targets, which indicated that these flavonoids possessed potential bioactivities in their interactions with the targets. The flavonoids in C. praecox exerted pharmacological activities in a multi-component, multi-target, and multi-pathway manner. The combined application of metabolomics and network pharmacology provides a theoretical basis for in-depth studies on the pharmacological effects and mechanisms of C. praecox.
Flavonoids/metabolism* ; Network Pharmacology ; Metabolomics/methods* ; Molecular Docking Simulation ; Calycanthaceae/chemistry* ; Tandem Mass Spectrometry ; Drugs, Chinese Herbal/chemistry*

Chimonanthus plants widely distributed in southern area of China, which have a long history of edibles and medicine. Phytochemical investigations have shown that Chimonanthus produced 143 non-volatile constituents, including alkaloids, flavonoids, terpenoids, coumarins and others, which exhibit significant anti-oxidant, anti-bacterial, anti-cancer, anti-inflammatory, antihyperglycemic, antihyperlipidemic and other biological activities. On the basis of systematic reviewing of literatures, this article overviews the non-volatile constituents and pharmacology of Chimonanthus from domestic and foreign over the last 30 years (until June 2018), and may provide a useful reference for the further development of Chimonanthus.
Animals ; Calycanthaceae ; chemistry ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; therapeutic use ; toxicity ; Humans ; Medicine, Chinese Traditional ; Phytochemicals ; chemistry ; pharmacology ; therapeutic use ; toxicity ; Phytotherapy

Scopolin (SC-1), scopoletin (SC-2) and isofraxidin (IS-1) are the main active constituents in Chimonanthi Radix. However, the in vivo metabolism of SC-1, SC-2 and IS-1 have not been comprehensively clarified. In this study, the in vivo metabolic profiles of these three coumarins in the rat plasma, urine and feces were analyzed. Ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry (UHPLC-QTOF-MS/MS) method was applied to characterize the prototypes and metabolites of SC-1, SC-2 and IS-1 in rat feces, urine, and plasma after intravenous administration. A total of 11 metabolites of the three parent compounds were tentatively identified. The main metabolic pathways were analyzed by identification of metabolites, and it was found that these three coumarins underwent multiple in vivo metabolic reactions including glucuronidation, sulfonation, isomerism and reduction. In this study, the analysis of metabolites of three coumarins basically demonstrated their in vivo metabolic process, providing basis for the further pharmacokinetics and pharmacological evaluations of SC-1, SC-2 and IS-1.
Animals ; Calycanthaceae ; chemistry ; Chromatography, High Pressure Liquid ; Coumarins ; metabolism ; pharmacokinetics ; Drugs, Chinese Herbal ; metabolism ; pharmacokinetics ; Rats ; Tandem Mass Spectrometry

In the present study, four new sesquiterpenoids, chimonols A-D (compounds 1-4), together with four known compounds (5-8) were isolated from the EtOAc extract of Chimonanthus praecox Link. The structures of these new compounds were elucidated on the basis of spectroscopic techniques (UV, IR, MS, and 1D and 2D NMR), and their absolute configurations were established by comparing experimental and calculated electronic circular dichroism (ECD) spectra. Compounds 1-8 were evaluated for antimicrobial activities and the minimum inhibitory concentrations (MICs) were determined by the broth microdilution method in 96-well culture plates. Compounds 1, 2, and 7 exhibited weak antibacterial effects for S. aureus (ATCC 6538), E. coli (ATCC 11775), and P. aeruginosa (ATCC 10145) with MIC values being 158-249 µg·mL. Compounds 3-7 showed activities against C. glabrata (ATCC 2001) and S. aureus (ATCC 43300) with MIC values being 128-197 µg·mL. Compounds 1-4 showed activity against S. aureus (ATCC 25923) with MIC values being 162-254 µg·mL. The present study provided a basis for future evaluation of these compounds as antibacterial agents.
Anti-Bacterial Agents ; chemistry ; isolation & purification ; pharmacology ; Calycanthaceae ; chemistry ; Escherichia coli ; drug effects ; Microbial Sensitivity Tests ; Molecular Structure ; Plant Extracts ; chemistry ; isolation & purification ; pharmacology ; Sesquiterpenes ; chemistry ; isolation & purification ; pharmacology ; Staphylococcus aureus ; drug effects

Glucose-6-phosphate dehydrogenase is main regulatory enzyme for pentose phosphate pathway. To amplify the core sequence of G6PDH gene from Chimonanthus praecox, the primers were synthesized, based on the conserved nucleotide sequence of other reported plant G6PDH genes. The specific primers were designed according to the major fragment. The full length cDNA of the G6PDH1 gene was isolated by the 3' and 5' rapid amplification of cDNA ends approach. Transcript levels of G6PDH1 isoform was measured by real-time quantitative RT-PCR in different tissues and in responds to cold treatment. The G6PDH1 subcellular localization, transmembrane domain, three-dimensional structure, and phylogenetic analysis were predicted by different software to analysis the bioinformatics of G6PDH1 protein. The G6PDH1 cDNA sequence was 2 011 bp in length and consisted of 1 551 bp Open Reading Frame (ORF) , encoding a protein of 516 amino acids. Expression analysis results in different tissues showed that G6PDH1 was primarily observed in flowers and roots, as opposed to the leaves and stems. Cold treatment experiments indicated that cold treatment caused a rapid increase in G6PDH1 expression in flowers within 12 h. The full-length cDNA of G6PDH1 and its expression analysis will play an important role for further study on cold stress responses in Ch. praecox.
Calycanthaceae ; chemistry ; classification ; enzymology ; genetics ; Cloning, Molecular ; Enzyme Stability ; Glucosephosphate Dehydrogenase ; chemistry ; genetics ; metabolism ; Models, Molecular ; Open Reading Frames ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism

This study was aimed to establish the HPLC fingerprints of the genus Chimonanthus leaves and compare the constituents distribution among five Chimonanthus species . The analysis was conducted on a C15 column (4.6 mm x 250 mm, 5 microm) with the mobile phase containing acetonitrile-water in gradient program: acetonitrile (B), 0-20 min, 6%-20%; 20-30 min, 20%-25%; 30-40 min, 25%-45%; 40-50 min, 45%-80%; 50-80 min, 80%-85%; 80-90 min, 85%-100%; 90-110 min, 100%. Flow rate was 0.8 mL x min(-1) and detection wavelength was 228 nm. Column temperature was set at 30 degrees C. The HPLC fingerprints of the five Chimonanthus species have been established. Ch. praecox, Ch. nitens, Ch. salicifolius, Ch. Zhejiangensis and Ch. grammatus have significant difference in constituents distribution and contents. Five standard substances as common compounds were confirmed in chromatography fingerprints. The method can be used as quality evaluation and classicfication of the genus Chimonanthus.
Calycanthaceae ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Plant Leaves ; chemistry

To explore anti-tumor active components of Chimonanthus salicifolius, the phytochemistry of the chloroform fraction from leaves extract was investigated by repeated silica gel column chromatography. Twelve compounds were isolated and their structures were identified by physicochemical properties and spectroscopic data analysis as 9-epi-blumenol C(1), blumenol C(2), (+)-dehydrovomifoliol (3), (+)-vomifoliol (4), robinlin (5), (-)-loliolide (6), isofraxidin (7), scopoletin (8), 6,7-dimethoxycoumarin (9), 6, 7, 8-trimethoxycoumarin (10), beta-sitostenone (11), and beta-stigmasterol(12). Compounds 1-6 belonging to nor-sesquiterpenoids were isolated from the family Calycanthaceae for the first time. Compound 1 was a new natural product. Compounds 7, 11 and 12 were obtained from this plant for the first time.
Antineoplastic Agents ; analysis ; isolation & purification ; Calycanthaceae ; chemistry ; Chloroform ; chemistry ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Plant Leaves ; chemistry

The vapour distillation was used to extract the volatile oil of Chimonanthus salicifolius with different storage time, determine the content of cineole in volatile oil by GC, to study the influence of storage time on the content of volatile oil and cineole of C. salicifolius. We found that the content of volatile oil in fresh herbs of C. salicifolius was 0.023 0 mL x g(-1), it was decreased to 0.020 0, 0.017 5 mL x g(-1) respectively after storing for 4, 12 months; the GC methodological study of precision, stability and repeatability, RSD < 2%, the average recovery rate was 99.50%, RSD 1.7%; the content of cineole in fresh volatile oil was 54.30%, it was increased to about 62% and remained stably with the time. Therefore, the content of volatile oil and cineole of C. salicifolius can change with the storage time; GC method for the determination of the content of cineole is accurate, reliable, specific and repeatable, it's suitable as a quality control method of C. salicifolius.
Calycanthaceae ; chemistry ; Chromatography, Gas ; Cyclohexanols ; analysis ; Drug Storage ; methods ; Drugs, Chinese Herbal ; analysis ; Monoterpenes ; analysis ; Oils, Volatile ; analysis ; Plant Oils ; analysis

The chemical composition of essential oil, which from the leaves of Chimonanthus grammatus obtained by hydrodistillation were analyzed by GC-MS, and their possible antibacterial properties were screened. According to the results from GC-MS analysis, fifty-three components comprising 99.99% of the essential oil were identified. The major components of essential oil were 3-(4, 8-dimethyl-3,7-nonadienyl)(E) -furan (13.1%), bornyl acetate (12.66%), and 6,6-dimethyl-3-methylene-bicyclo[3.1.1] heptane (7.06%), etc. Antibacterial activity of essential oil was employed by two complementary test systems of disc diffusion and MIC/ MBC tests, which showed obviously antibacterial activity against all of the tested bacteria.
Anti-Bacterial Agents ; pharmacology ; Calycanthaceae ; chemistry ; Gas Chromatography-Mass Spectrometry ; methods ; Oils, Volatile ; analysis ; pharmacology ; Plant Leaves ; chemistry