A new coumarin and a new flavonoid from Ochrocarpus longifolius.

A new coumarin (1) and a new flavonoid (2) were isolated from the air-dried flower buds of Ochrocarpus longifolius, together with ten known compounds (3-12). The structures of two new compounds were established by 1D and 2D NMR and MS data. In addition, the new compound 2 showed significant proliferation inhibitory activity on Eca-109 and MGC-803 cells. The results of this study may enrich the diversity of compounds from O. longifolius and provide a basis for further research on its natural products and pharmacological activities.

[Study on chemical components of Hypericum himalaicum and mechanism of anti-inflammatory based on network pharmacology and molecular docking technology].

The chemical constituents of ethyl acetate from Hypericum himalaicum were isolated by silica gel column chromatography, gel column chromatography, and high-performance liquid chromatography. The structure of the isolated compounds was identified by modern spectral techniques(NMR, MS, IR, and UV), and the potential anti-inflammatory targets and action pathways were analyzed and predicted by network pharmacology and molecular docking methods.Ten compounds were isolated from H. himalaicum and identified as 5,9,11-trihydroxy-3,3-dimethyl-3H,8H-benzo[6,7][1,4]dioxepino[2,3-f]chromen-8-one(1), betulinic acid(2), demethyltorosaflavone C(3), kaempferol(4), quercetin(5), hyperwightin B(6), toxyloxanthone B(7), 1,7-dihydroxy-xanthone(8), emodin(9), and 1,7-dihydroxy-4-methoxy-xanthone(10). Among them, compound 1 was a new compound, and compounds 2-10 were isolated from H. himalaicum for the first time. Network pharmacology screened 60 key anti-inflammatory targets. By acting on TNF, AKT1, CASP3, and other key targets, involving PI3K-AKT signaling pathway, IL-17 signaling pathway, VEGF signaling pathway, MAPK signaling pathway, and other signaling pathways, and phosphorylation, cell migration and movement, protein tyrosine kinase, and other biological processes were regulated to achieve anti-inflammatory effects. The results of molecular docking show that the above components have good binding properties with the core targets.

Acute and 13 weeks subchronic toxicological evaluation of the flavonoid-rich extract of Sophora flavescens.

The roots of Sophora flavescens have a long history of use in Chinese medicine for the treatment of various medical conditions. Flavonoids from the ethyl acetate extract of S. flavescens have shown anti-inflammatory, anticancer, and antidiabetic properties. The objective of this study was to evaluate the toxicological profile of a flavonoid-rich extract of S. flavescens (SFEA). We conducted acute and sub-chronic oral toxicity studies of SFEA in Kunming (KM) mice and Sprague-Dawley (SD) rats. Acute oral administration of 9.0 g/kg SFEA did not result in mortality, clinical signs of toxicity, or abnormal changes in the body weight or food consumption patterns. No significant changes in hematological, blood biochemical, or histopathological parameters were observed. A 13-week sub-chronic toxicity study was conducted in SD rats; the rats were orally administrated with various doses of SFEA (in mg/kg): 0 (control), 40, 80, 400, 800, and 1200. Mortality, clinical signs, or treatment-related changes in body weight, food consumption, hematological parameters, blood biochemical parameters, organ weights, or histopathological parameters were not observed. We found that SFEA is practically nontoxic to KM mice at a dose of 9.0 g/kg and that the no-observed-adverse-effect-level (NOAEL) of SFEA in SD rats is greater than 1200 mg/kg.

Chemistry, Bioactivity, and Prediction of the Quality Marker (Q-Marker) of Ferula Plants in China: A Review.

The genus of Ferula belongs to the family Apiaceae, and many Ferula plants are used as traditional Chinese medicines. Ferula plants were initially identified as early as the "Newly Revised Materia Medica" written in the Tang Dynasty (AD 659), and several of them are also recognized as the traditional medicines of the Uygur, Kazakh, and Mongolian. Ferula plants are distributed in China, Russia, India, Africa, Central Asia, and other places. Currently, the chemical components derived from Ferula plants are mainly coumarins, sesquiterpenes, and volatile oils. Ferula plants can exhibit diverse pharmacological activities such as anti-allergy, analgesia, relieving cough, anticoagulation, and anti-tumor. Therefore, this article summarized the domestic research conducted on the genus Ferula, appropriately combines the research status of the foreign genus Ferula, and describes the chemical composition, biological activity, toxicity issues, and Q-marker prediction. In addition, all the related studies about the genus Ferula are summarized by analyzing the various databases such as CNKI, Wanfang data, PubChem and SciFinder.

Penpolonin A-E, cytotoxic α-pyrone derivatives from Penicillium polonicum.

Five new α-pyrone derivatives, named penpolonin A-E (1-5), together with two known compounds (6-7) were acquired from the endophytic fungus Penicillium polonicum isolated from the roots of Camptotheca acuminata Decne. Their structures were established by combination of NMR and HRESIMS data and the absolute configurations of 1-5 were determined by NMR calculations and comparison of experimental and calculated ECD data. Compounds 3 and 7 exhibited moderate cytotoxicity against Hep-2, TU212 human laryngeal cancer cells with IC50 values ranging from 31.6 to 45.1 µg/ml, compound 4 showed weak cytotoxicity against the Hep-2 and TU212 cell lines with IC50 values of 69.2 and 68.7 µg/ml.

New flavonoids from the roots of Sophora davidii (Franch.) Skeels and their glucose transporter 4 translocation activities.

Five new flavanones, davidones A-E (1-5), one new isoflavonoid, cyclolicoisoflavones A3 (8), together with seven known compounds were isolated from the petroleum ether and the ethyl acetate fractions of the roots of Sophora davidii (Franch.) Skeels. The structures of new compounds were established by 1D and 2D NMR and MS data. The absolute configuration of 1-5 was assigned by NMR calculations and comparing its experimental and calculated ECD spectra. Flavanones were the main active principles responsible for the glucose transporter 4 (GLUT-4) translocation activities of SD-PE and SD-EtOAc. Compounds 1-7 and acacetin (12) promoted GLUT-4 translocation by the range of 1.35-3.00 folds, respectively.

A biflavonoid-rich extract from Selaginella moellendorffii Hieron. induces apoptosis via STAT3 and Akt/NF-κB signalling pathways in laryngeal carcinoma.

Selaginella moellendorffii Hieron. (SM), a perennial evergreen plant, has been used in the treatment of acute infectious hepatitis, thoracic and hypochondriac lumbar contusions, systemic oedema and thrombocytopaenia. However, the role of a biflavonoid-rich extract from SM (SM-BFRE) in anti-larynx cancer has rarely been reported. In this study, the in vitro and in vivo anti-laryngeal cancer activity and potential mechanisms of SM-BFRE were investigated. An off-line semipreparative liquid chromatography-nuclear magnetic resonance protocol was carried out to determine six biflavonoids from SM-BFRE. In vitro, MTT assay revealed that SM-BFRE inhibited the proliferation of laryngeal carcinoma cells. A wound healing assay indicated that SM-BFRE suppressed the migration of laryngeal cancer cells. Hoechst 33 258 and Annexin V-FITC/PI double staining assays were performed and verified that SM-BFRE induced apoptosis in laryngeal carcinoma cells. The Hep-2 bearing nude mouse model confirmed that SM-BFRE also exhibited anticancer effect in vivo. In addition, Western blot analysis demonstrated that SM-BFRE exerted its anti-laryngeal cancer effect by activating the mitochondrial apoptotic pathway and inhibiting STAT3 and Akt/NF-κB signalling pathways. All results suggested that SM-BFRE could be considered as a potential chemotherapeutic drug for laryngeal cancer.

Phenolic Constituents with Glucose Uptake and GLUT4 Translocation Bioactivities from the Fruits of Cordia dichotoma.

From the fruits of Cordia dichotoma, 11 new phenolic compounds, dichotomins A-K, were isolated, together with 19 known compounds. Through the analysis of detailed NMR data and HRESIMS data, the planar structures of all compounds were confirmed. Using NMR calculations, the absolute configuration of dichotomins A-K was elucidated by comparing their observed and computed electronic circular dichroism (ECD) spectra. Dichotomin H (8) and dichotomin I (9) were determined as two pairs of enantiomers. The enantiomers of compounds 8 and 9 were separated using chiral-phase high-performance liquid chromatography (HPLC), and the stereostructure of each enantiomer was determined by similarly calculating the ECD. Compounds 3, 5, 7, 17, 18, 23-25, and 27-30 increased glucose uptake by 1.04- to 2.85-folds at concentrations of 30 µg/mL. Further studies revealed that compounds 3 and 5 had a moderate effect on glucose transporter 4 (GLUT4) translocation activity in L6 cells. At 30 µg/mL, compound 3 significantly enhanced AMPK phosphorylation and GLUT4 expression. As a whole, compound 3 has the potential to be a drug candidate for the treatment of type 2 diabetes mellitus (T2DM).

Evaluation of toxicological safety and quality control of Luobufukebiri pill.

ETHNOPHARMACOLOGICAL RELEVANCE: The Luobufukebiri pill is one of the characteristic medicines of Uygur nationality in Xinjiang. It has the effect of warming and tonifying the brain and kidney, benefiting the heart and filling the essential functions, mainly used to treat impotence, depression, spermatorrhea, premature ejaculation, bodily weakness, emaciation, and neurasthenia. AIM OF THE STUDY: This study evaluated the toxicology and developed a quality control protocol of Luobufukebiri pill to ensure its safety and effectiveness in clinical applications. MATERIALS AND METHODS: Acute toxicity in mice was studied by the maximum-dose method, and the toxic reactions in mice were observed within two weeks. In the study of Sub-chronic toxicity, SD rats were randomized into four groups: three drug groups which were treated with 8.00, 2.67, and 0.80 g/kg of Luobufukebiri pill, respectively, and one control group which was treated with the same volume of distilled water. Subsequently, at 30 days of medication and 30 days of drug withdrawal, the hematologic indexes, biochemical indexes, organ coefficient, and pathological sections of main organs were detected, respectively. According to the prescription, the contents of 8 active components in the pill were quantified simultaneously. The chromatographic conditions were as follows: Stepwise gradient elution was carried out using 0.1% formic acid (solvent A) and acetonitrile (solvent B), 0-8 min, 80% → 60% B; 8-25 min, 60% → 25%B. The flow rate was 1.0 mL/min, the column was maintained at 25 °C, and the injected sample volume was 10 µL. RESULTS: The acute toxicity experiment documented a large dose of Luobufukebiri pill had no significant effect on organ and body weight and did not cause apparent damage to parenchymal organs. At Sub-chronic toxicity, the behavior of rats was as normal as the control group. There were some differences in hematologic indexes, serum biochemical indexes, and organ coefficient tests between the drug and control groups, but they had no toxic significance. No obvious pathological changes were observed in the pathological sections of major organs. In conclusion, this study demonstrated that the clinical dose of Luobufukebiri pill was far less than its toxic dose, and it had reliable safety. The contents of eight index components of Luobufukebiri pill were measured. All calibration curves exhibited good linearity with correlation coefficients better than 0.9997. The relative standard deviations of precision, reproducibility, stability, and recovery were less than 2.0%, demonstrating the stability and reliability of the method. CONCLUSIONS: This study further confirmed the safety of Luobufukebiri pill in clinical practice. A rapid, accurate, and convenient RP-HPLC-PDA detection method has been developed for the simultaneous detection of eight active compounds in the pharmaceutical samples of Luobufukebiri pill. This study provided a reference for the safety and enhancement of the quality standards of Luobufukebiri pill.

Traditional Uses, Chemistry, Pharmacology, Toxicology and Quality Control of Alhagi sparsifolia Shap.: A Review.

Alhagi sparsifolia Shap. (Kokyantak) is a ethnic medicine used in the Uyghur traditional medicine system for the treatment of colds, rheumatic pains, diarrhea, stomach pains, headaches, and toothaches, in addition to being an important local source of nectar and high-quality forage grass, and playing a crucial role in improving the ecological environment. Currently, approximately 178 chemical constituents have been identified from A. sparsifolia, including flavonoids, alkaloids, phenolic acids, and 19 polysaccharides. Pharmacological studies have already confirmed that A. sparsifolia has antioxidant, anti-tumor, anti-neuroinflammatory effects, hepatoprotective effects, renoprotective effects and immune regulation. Toxicological tests and quality control studies reveal the safety and nontoxicity of A. sparsifolia. Therefore, this paper systematically summarizes the traditional uses, botany, phytochemistry, pharmacology, quality control and toxicology of A. sparsifolia, in order to provide a beneficial reference of its further research.

Methylgerambullin derived from plant Glyccsmis pentaphylla (Retz) correa. Mediates anti-hepatocellular carcinoma cancer effect by activating mitochondrial and endoplasmic reticulum stress signaling and inhibiting AKT and STAT3 pathways.

Hepatocellular carcinoma (HCC) is one of the most common fatal malignant tumors. Glycosmis pentaphylla is used by traditional medical practitioners worldwide to treat various diseases. We isolated and identified a chemical component with potential anti-hepatocellular carcinoma (HCC) effects. Methylgerambullin is a sulfur containing amine and has significant antihepatoma activity in vitro and in vivo. Methylgerambullin was significantly cytotoxic to HCC cells and induces apoptosis in HCC cells. In addition, methylgerambullin is able to inhibit the growth of transplanted tumors in nude mice without significant toxicity. Regarding the anti-cancer mechanism of methylgerambullin, treatment with methylgerambullin increased the expression of caspase-3, caspase-9 and Bax in vitro and in vivo and reduce the expression of B-cell lymphoma-2 (Bcl-2). Simultaneously, methylgerambullin can also affect ERS-related proteins, inhibit Protein Kinase B (Akt) activity, cause dephosphorylation of downstream Bad, and inhibit the expression of the Signal Transducer and Activator of Transcription 3 (STAT3) protein to inhibit HCC cells proliferation. Overall, these results suggest that methylgerambullin can inhibit HCC cells proliferation by inducing mitochondrial apoptosis, activating ERS signaling pathways and inhibiting the Akt and STAT3 pathways.

Antidiabetic effect of a flavonoid-rich extract from Sophora alopecuroides L. in HFD- and STZ- induced diabetic mice through PKC/GLUT4 pathway and regulating PPARα and PPARγ expression.

HEADINGS ETHNOPHARMACOLOGICAL RELEVANCE: Sophora alopecuroides L. is a traditional ethnopharmacological plant, which is widely used in traditional Chinese medicine and Mongolian and Uighur medicine to ameliorate "thirst disease". AIM OF THE STUDY: This study aimed to investigate the antidiabetic activities and mechanisms of a flavonoid-rich extract from Sophora alopecuroides L. (SA-FRE) both in vivo and vitro. MATERIALS AND METHODS: The main six chemical constituents of SA-FRE were elucidated based on an off-line semi-preparative liquid chromatography nuclear magnetic resonance (LC-NMR) protocol. Myc-GLUT4-mOrange-L6 cell models and mouse model with diabetes induced by high-fat diet combined with STZ injection were respectively adopted to investigate the antidiabetic effects of SA-FRE both in vitro and vivo. RESULTS: In vivo, 4-week treatment of SA-FRE ameliorated hyperglycemia, dyslipidemia, and insulin resistance in diabetic mice. Mechanically, SA-FRE regulated PPARα and PPARγ expression in white adipose tissue (WAT) and liver, thereby ameliorating dyslipidemia. Moreover, SA-FRE increased the phosphorylation of PKC and further stimulated the GLUT4 expression in WAT and skeletal muscle, thus increasing the glucose utilization in vivo. In vitro, 50 µg/mL SA-FRE increased GLUT4 translocation to about 1.91-fold and glucose uptake to 1.82-fold in L6-myotubes. SA-FRE treatment increased the GLUT4 expression at both gene and protein levels. Furthermore, only Gö6983, a PKC inhibitor, reversed the SA-FRE-induced GLUT4 translocation and expression at the gene and protein levels. CONCLUSIONS: Generally, SA-FRE ameliorated hyperglycemia, dyslipidemia, and insulin resistance partly through activating PKC/GLUT4 pathway and regulating PPARα and PPARγ expression.

Anti-esophageal Cancer Effect of Corilagin Extracted from Phmllanthi Fructus via the Mitochondrial and Endoplasmic Reticulum Stress Pathways.

HEADINGS ETHNOPHARMACOLOGICAL RELEVANCE: Corilagin (ß-1-O-galloyl-3,6-(R)-hexahydroxydiphenoyl-d-glucose) is a tannin isolated from the traditional ethnopharmacological plant Phmllanthi Fructus, which is widely used in not only traditional Chinese medicine but also tropical and subtropical medicine to ameliorate various diseases. AIM OF THE STUDY: This study was designed to isolate the potential anti-esophageal cancer (EC) component corilagin from Phmllanthi Fructus and explain its anti-EC mechanism. MATERIALS AND METHODS: Corilagin was isolated from Phmllanthi Fructus by extraction and chromatographic procedures, and its anti-esophageal cancer effect was evaluated by in vitro and in vivo experiments. In vitro experiments included MTT analysis, flow cytometry, and the Transwell assay and were used to observe corilagin-mediated inhibition of EC cell growth. Western blotting was used to analyze the apoptotic pathway of EC cells. In vivo experiments used tumor-bearing nude mice to evaluate the antitumor effect of corilagin, and its potential mechanism was explored by Western blotting. RESULTS: Corilagin showed significant anti-EC activity in vitro and in vivo. Corilagin was significantly cytotoxic to EC cells and induced apoptosis in EC cells. Corilagin induced G0/G1 phase arrest by altering key G0/G1 cell cycle regulatory markers and significantly reducing the migration of EC cells and the number of cells in a time- and dose-dependent manner. Additionally, corilagin inhibited the growth of transplanted tumors in nude mice without significant toxicity. Regarding the anticancer mechanism of corilagin, the results showed that corilagin inhibited esophageal cancer progression by activating mitochondrial and endoplasmic reticulum stress signaling pathways. CONCLUSIONS: Corilagin shows significant anti-EC activity in vitro and in vivo. The mechanism of the anti-EC activity of corilagin may be due to activating mitochondrial and endoplasmic reticulum stress signaling pathways.

Study on evaluation of toxicology and quality control of Yimusake tablet.

ETHNOPHARMACOLOGICAL RELEVANCE: The Yimusake tablet (YMSK-T) is a type of Xinjiang Uygur Medicine, which affects curing diseases of impotence and premature ejaculation. It has remarkable pharmacological effects that mainly involve improving the number and shape of smooth muscle cells in the corpus cavernosum and enhancing the relaxation and contraction function of corpus cavernosum smooth muscle. AIM OF THE STUDY: The YMSK-T prescription, which consists of 11 traditional herbs, has significant pharmacological effects, however the evaluation of toxicology and quality control of the preparation has not yet been reported. Therefore, in this study, we evaluated the toxicology and quality control of YMSK-T to ensure its safety and effectiveness in clinical applications. MATERIALS AND METHODS: Male rats were divided into three groups and were given continuous gavage administration of high, medium and low concentrations of YMSK-T. To determine hematopoietic parameters, orbital blood was collected at regular intervals. At termination of the experiment, rats were dissected for histopathological examination. According to the function of the prescription medicinal materials, seven active components were selected for content determination under the same chromatographic condition of using 0.2% aqueous phosphoric acid (solvent A) and acetonitrile (solvent B) with a 40 min post time: 0-13 min, 20% →30% B; 13-26 min, 30% →72% B; 26-38 min, 72% →92% B; 38-40 min, 92% →96% B. The column was maintained at 25 °C and the total sample injection was 10 µL. RESULTS: Our data showed that using a large dose (400X the dosage used in humans) of YMSK-T resulted in myocardium and liver damage, and eventually death of the rats. At sub-chronic toxicity, no significant differences were observed among indexes about relative organ weight, hematology, serum biochemistry and histopathological examination, and rats behaved normally. Our results also demonstrated that the YMSK-T dosage used was not toxic in the normal range. The linearity of each component was sufficient (correlation coefficients>0.9997). Moreover, the relative standard deviations of precision, repeatability, stability, and recovery were less than 2.0%, which showed that the method for determination of content was stable and reliable. CONCLUSIONS: YMSK-T has been found to be relatively safe in a rat model, and the method of content determination can be used for quality control of YMSK-T. Toxicology and quality control studies indicated that, the drug is safe and effective for clinical application.