He-Wei Granule enhances anti-tumor activity of cyclophosphamide by changing tumor microenvironment.

Objective: He-Wei Granule (HWKL) is a modern product derived from the modified formulation of traditional Chinese medicine Banxia Xiexin Decoction (BXD), which remarkedly enhanced the anti-proliferation activity of cyclophosphamide (CTX) on HepG2 and SGC-7901 cell lines in vitro in our previous research. The aim of the study was to investigate the synergistic effects of HWKL and CTX using a transplanted H22 hepatocellular carcinoma mouse model. Methods: The CTX-toxic-reducing efficacy of HWKL was evaluated by hematology indexes, organ indexes and marrow DNA detection. To investigate the underlying mechanisms, histopathology test, immunohistochemistry test and TUNEL staining were conducted. The efficacy of HWKL on the micro-vessel density (MVD) in tumor tissue was also evaluated by measuring CD34 level. Results: High dose HWKL (6.75 g/kg) markedly attenuated CTX-induced hepatotoxicity and myelosuppression while significantly enhanced CTX anticancer efficacy in vivo. Further mechanism investigation suggested that high dose HWKL significantly increased cleaved Caspase 3 level and promoted apoptosis in tumor tissue by up-regulating Bax expression and down-regulating Bcl-2 and FasL expressions. Compared with CTX alone group, the decrease in LC-3B and Beclin 1 levels suggested that the autophagy in H22 carcinoma was significantly inhibited with addition of high dose HWKL. ELISA assay results indicated that the autophagy inhibition was achieved by decreasing p53 expression, blocking PI3K/AKT/mTOR pathway and recovering Th1/Th2 cytokine balance. In addition, CD34 and EGFR immunohistochemistry assay suggest that high dose HWKL could significantly decrease micro-vessel density (MVD) and inhibit angiogenesis in H22 carcinoma. Conclusion: It can be concluded that high-dose HWKL enhanced CTX efficacy by promoting apoptosis, inhibiting autophagy and angiogenesis in tumor tissue while significantly alleviated CTX-induced toxicity, and could be applied along with CTX in clinical treatment as a supplement agent.

Antioxidant, Anti-Inflammatory, and Antidiabetic Activities of Bioactive Compounds from the Fruits of Livistona chinensis Based on Network Pharmacology Prediction.

In this study, a chemical investigation on the fruits of Livistona chinensis (FLC) led to the isolation and identification of 45 polyphenols and 5 alkaloids, including two new compounds (Livischinol (1) and Livischinine A (46)), an undescribed compound (47) and 47 known compounds. FLC was predicted with novel potential antidiabetic function by collecting and analyzing the potential targets of the ingredients. Compound 32 exhibited significant α-glucosidase inhibitory activity (IC50 = 5.71 µM) and 1, 6, and 44 showed the PTP1B inhibitory activity with IC50 values of 9.41-22.19 µM, while that of oleanolic acid was 28.58 µM. The competitive inhibitors of PTP1B (compounds 1 and 44) formed strong binding affinity, with catalytic active sites, proved by kinetic analysis, fluorescence spectra measurements, and computational simulations, and stimulated glucose uptake in the insulin-resistant HepG2 cells at the dose of 50 µM. In addition, FLC was rich in antioxidant and anti-inflammatory bioactive compounds so that they could be developed as nutraceuticals against diabetes.

Characterization of antioxidant, α-glucosidase and tyrosinase inhibitors from the rhizomes of Potentilla anserina L. and their structure-activity relationship.

Five new flavonoids (1-5), along with 25 known compounds, were isolated from the rhizomes of Potentilla anserina L. and their structures were identified using spectroscopic and chemical evidence. The extract, all fractions, and all isolated compounds were evaluated for their antioxidant, α-glucosidase, and tyrosinase inhibitory activities, and their structure-activity relationship was interpreted. The biflavanols and quercetin-3-O-α-l-rhamnopyranoside-2″-gallate (14) exhibited significant antioxidant and α-glucosidase inhibition activities. In this study, anti-tyrosinase activity and its mechanism of active compounds (potenserin C (4), potenserin D (5), and quercetin-3-O-α-l-rhamnopyranoside-2″-gallate (14)) were explored by a combination of computational simulations and kinetic studies. Kinetic studies indicated that potenserin C (4) and quercetin-3-O-α-l-rhamnopyranoside-2″-gallate (14) inhibited tyrosinase in a competitive manner, whereas potenserin D (5) acted in a reversible noncompetitive manner. The molecular docking result indicated that the substitution of the glucose moiety with galloyl and the presence of 3', 4', 5'-OH in flavonoid aglycones played a crucial role for the tyrosinase inhibiting effect. Moreover, the presence of biflavanols increased the activity against tyrosinase because of strong hydrogen binding, π-alkyl binding, and electrostatic interaction. Thus, the presented experiments developed several new lead compounds that could act as antioxidants and α-glucosidase inhibitors. Furthermore, biflavanols and quercetin-3-O-α-l-rhamnopyranoside-2″-gallate played important roles in the anti-browning activity during food processing.

Design and Synthesis of Polymer Prodrugs for Improving Water-Solubility, Pharmacokinetic Behavior and Antitumor Efficacy of TXA9.

PURPOSE: TXA9, a novel cardiac glycoside, has a potent anti-proliferative effect against A549 human lung cancer cells, however, possesses a poor water-solubility and a rapid metabolic rate in vivo which limited the further development of TXA9. To overcome the shortcomings of TXA9, four polymer prodrugs of TXA9 were designed and synthesized. METHODS: Poly (ethylene glycol) monomethyl ether (mPEG) and α-tocopherol polyethylene glycol succinate (TPGS) were applied to modify TXA9 via carbonate ester and glycine linkers respectively to obtain four polymer prodrugs. The water-solubility and stability of prodrugs were studied in vitro while their pharmacokinetic behaviors and antitumor activity were investigated in vivo. RESULTS: The water-solubility of TXA9 was obviously increased and prodrugs with glycine linkers showed a better stability in rat plasma. Their pharmacokinetic investigation found that the t1/2 and AUC0-∞ of TPGS-Gly-TXA9 was increased by 80- and 9.6-fold compared with that of TXA9, which was more superior than the other three prodrugs. More importantly, the tumor inhibition rate of TPGS-Gly-TXA9 (43.81%) on A549 xenograft nude mice was significantly increased compared with that of TXA9 (25.26%). CONCLUSION: The above results suggested that TPGS-Gly-TXA9 possessed better antitumor efficiency than TXA9 and could be further investigated as an anti-cancer agent.

Antioxidant and α-Glucosidase Inhibitory Activities Guided Isolation and Identification of Components from Mango Seed Kernel.

In the present study, petroleum ether, dichloromethane, ethyl acetate, and n-butanol fractions of mango seed kernel exhibited different degrees of antioxidant and α-glucosidase inhibitory activity. Thus, quantitative and qualitative analysis of the petroleum ether fraction was conducted by GC-MS. Among identified components, four unsaturated fatty acids had never been reported in natural products before, together with 19 known components. In addition, 17 compounds were isolated and elucidated from other active fractions. Compounds 2, 9, 15, and 17 were isolated for the first time from Mangifera genus. Compounds 1 and 2 exhibited prominent DPPH radical scavenging and α-glucosidase inhibitory effects. In order to further explore their mechanism of α-glucosidase inhibition, their enzyme kinetics and in silico modeling experiments were performed. The results indicated that 1 inhibited α-glucosidase in a noncompetitive manner, whereas 2 acted in a competitive manner. In molecular docking, the stability of binding was enhanced by π-π T-shaped, π-alkyl, π-π stacked, hydrogen bond, and electrostatic interactions. Thus, compounds 1 and 2 were determined to be new potent antioxidant and α-glucosidase inhibitors for preventing food oxidation and enhancing hypoglycemic activity.

LC-MS/MS method for simultaneous determination of famitinib and its major metabolites in human plasma.

AIM: To establish and validate an ultra-high-performance liquid chromatography-tandem mass spectrometry method for the rapid and simultaneous determination of famitinib and its metabolites in human plasma. RESULTS: All analytes demonstrated good correlation coefficients (R2 > 0.99), and the LLOQ was 0.05 ng/ml. The inter- and intraday accuracy and precision, as well as the stability of the samples, met the requirements of the US FDA. The extraction recovery and the matrix effect ranged from 87.58 to 116.06% and from 84.57 to 120.53%, respectively. CONCLUSION: The assay was successfully validated and applied to gastroenteropancreatic neuroendocrine tumor patients, and the assay may be used as a valuable tool in the clinic to determine the drug concentration of famitinib in the plasma of solid tumor patients.

Schisandra chinensis extract decreases chloroacetaldehyde production in rats and attenuates cyclophosphamide toxicity in liver, kidney and brain.

ETHNOPHARMACOLOGICAL RELEVANCE: Schisandra chinensis (Turcz.) Baill (S. chinensis) has been used for thousands years in China, and is usually applied in treatment of urinary tract disorders and liver injury. S. chinensis extract (SCE) has board protective effects on liver, kidney and nervous system. Schisandra lignans are generally considered as the bioactive components of SCE. AIM OF THE STUDY: To investigate the pharmacokinetic herb-drug interactions (HDIs) between SCE and cyclophosphamide (CTX). To evaluate the protective effects of SCE against CTX induced damage in rat liver, kidney and brain. MATERIALS AND METHODS: The pharmacokinetic HDIs between SCE and CTX were investigated by determining plasma concentrations of CTX and three metabolites, namely 4-ketocyclophosphamide (4-Keto), 2-dechloroethylcyclophosphamide (DCCTX) and carboxyphosphamide (CPM) using a previously developed UPLC-MS/MS method. To evaluate the protective effects of SCE pretreatment, toxicity and oxidation stress assessments along with histology investigations were carried out in rat liver, kidney and brain. RESULTS: The equimolar produced metabolite DCCTX was chosen to reflect chloroacetaldehyde (CAA, a toxic metabolite of CTX) production in rats. Single-dose pretreatment of SCE significantly reduced CAA production and decreased the Cmax and AUC0-24h of DCCTX by 69% and 49% respectively (P < 0.05). After pretreated with SCE for 7 consecutive days, the Cmax and AUC0-24h of DCCTX were still decreased (-25% and -37%, P < 0.05) when compared with CTX alone group. Parallel toxicity and oxidation stress investigations showed that single-dose SCE pretreatment significantly decreased plasma BUN and Cr levels (-12% and -46%, respectively) and reduced liver AST activity (-32%). Moreover, SCE pretreatment potently increased the brain GSH content by 7.8-fold, and reduced MDA levels in rat liver, kidney and brain by 39%, 28% and 31%, respectively (compared with CTX alone group). The protective effects of SCE were also supported by histological observations. CONCLUSION: Our experiment results suggest that S. chinensis may find use as a complementary medicine in CTX treatment.

Time- and NADPH-Dependent Inhibition on CYP3A by Gomisin A and the Pharmacokinetic Interactions between Gomisin A and Cyclophosphamide in Rats.

The traditional Chinese medicine Schisandra chinensis has remarkable protective effects against chemical-induced toxicity. Cyclophosphamide (CTX), in spite advances in chemotherapy and immunosuppressive regimes, is prone to cause severe toxicity due to its chloroacetaldehyde (CAA) metabolite produced by CYP3A. Our previous study identified that S. chinensis extract (SCE) co-administration potently decreased CAA production and attenuated liver, kidney and brain injuries in CTX-treated rats. Gomisin A (Gom A) is proved to be one of the most abundant bioactive lignans in S. chinensis with a significant CYP3A inhibitory effect. To find out whether and how Gom A participated in the chemoprevention of SCE against CTX toxicity, the Gom A-caused CYP3A inhibition in vitro as well as the pharmacokinetic interactions between Gom A and CTX in vivo were examined in this study. Using human liver microsomes, a reversible inhibition assay revealed that Gom A was a competitive inhibitor with a KI value of 1.10 µM, and the time- and NADPH-dependent CYP3A inhibition of Gom A was observed in a time-dependent inhibition assay (KI = 0.35 µM, kinact = 1.96 min-1). Hepatic CYP3A mRNA expression experienced a significant increase in our rat model with Gom A administration. This explained why CAA production decreased in the 0.5 h- and 6 h-pretreatment rat groups while it increased in the 24 h- and 72 h-pretreatment groups, indicating a bidirectional effect of Gom A on CYP3A-mediated CTX metabolism. The present study suggested that Gom A participates like SCE in the pharmacokinetic intervention of CTX by blocking CYP3A-mediated metabolism and reducing CAA production, and thus plays an important role in the chemopreventive activity of S. chinensis against CTX toxicity, in addition to the previously recognized protective effects. Also, the combined use of S. chinensis preparation or other drugs containing Gom A as the main component with CTX needed to be addressed for better clinical intervention.

Assessment of anti-diabetic activity of the aqueous extract of leaves of Astilboides tabularis.

ETHNOPHARMACOLOGICAL RELEVANCE: Astilboides tabularis has a long history of usage as the functional food with the effect of adjunctive therapy of diabetic in northeast of China. The present study was undertaken to assess anti-diabetic activity of the aqueous extract of leaves of A. tabularis and provided experimental evidence for the clinical usage of A. tabularis in the treatment of diabetes mellitus. MATERIALS AND METHODS: The aqueous extracts of leaves of A. tabularis (EAT) were orally administered at 300 and 600mg/kg dose to epinephrine-induced and alloxan-induced diabetes mice. Hypoglycemic effects, change in body weight, water intake, food intake, lipid profile and lipid peroxidant were assessed. Furthermore, the main chemical components of EAT were isolated and high performance liquid chromatography (HPLC) analysis was employed to identify the phytochemical composition of the plant extract. The finally, total flavonoids content and total phenolic content were quantified by colorimetric assay. RESULTS: EAT showed a significant reduction in blood glucose level at both 300 and 600mg/kg (ig). In addition, EAT also exhibited a profound antioxidant effect for increasing SOD and GSH-px activity and decreasing TBARS activity at both low and high dose. Furthermore, EAT also could significantly decrease the contents of TC and TG. The 8 main constituents of EAT are also isolated and characterized and the content of total flavonoid and total phenolic are 22% and 25% in EAT respectively. CONCLUSION: The present investigation revealed that EAT possess potent anti-diabetic activity and polyphenol compounds may be the main active ingredients.