Network Pharmacology-Based Analysis on the Curative Effect of Kunxian Capsules against Rheumatoid Arthritis.

Kunxian capsules (KCs), a Chinese patent medicine, have been clinically proven to be effective in the treatment of rheumatoid arthritis (RA). However, the chemical profile of KC remains to be characterized, and the mechanism underlying the protective effect against RA is yet to be elucidated. Here, a network pharmacology-based approach was adopted, integrated with the chemical profiling of KC by UHPLC-Q-TOF/MS. As a result, a total of 67 compounds have been identified from KC extract, among which 43 were authenticated by comparison to the mass spectrum of standard chemicals. ADME behaviors of the chemical constituents of KC were predicted, resulting in 35 putative active ingredients. Through target prediction of both active ingredients of KC and RA and PPI analysis, core targets were screened out, followed by biological process and related pathway enrichment. Then, a TCM-herb-ingredient-target-pathway network was constructed and a multicomponent, multitarget, and multipathway synergistic mechanism was proposed, providing an information basis for further investigation. The active pharmaceutical ingredients included mainly terpenoids (such as triptolide and celastrol), sesquiterpene pyridines (such as wilforgine and wilforine), and flavonoids (such as icariin, epimedin A, B, and C, and 2″-O-rhamnosylicariside II).

The effects of Salvia przewalskii total phenolic acid extract on immune complex glomerulonephritis.

CONTEXT: Salvia przewalskii Maxim. (Lamiaceae) is a Chinese herbal medicine that has long been used for the treatment of cardiovascular disease. OBJECTIVE: The study investigated the therapeutic efficacy of S. przewalskii total phenolic acid extract (SPE) on immune complex glomerulonephritis (ICG) in rats. MATERIALS AND METHODS: Sixty-two Wistar rats were randomized into six groups. ICG was induced in all groups except normal control group. SPE was administered intragastrically at 24 h intervals for 40 consecutive days. Urine protein (UP), total serum protein (TSP), serum albumin (SA), serum cholesterol (SC) and serum urea nitrogen (SUN) were measured one day before, on day 20 and 40 after SPE administration. On day 40 after SPE administration, the kidneys were removed and prepared into pathologic sections. In addition, kidney wet mass was measured for calculating the kidney wet mass coefficient (KWMC). RESULTS: UP excretion was reduced significantly on day 20 after SPE administration in all three SPE groups as compared with that in medium group, and this effect was observable continuously until 40 days after SPE administration. Compared with medium group, TSP and SA were increased in all three SPE groups after 40 days treatment, while SC and SUN were decreased. KWMC was decreased significantly in 100 mg/kg SPE group after 40 days treatment compared with that in medium group. Histopathologic analyses showed that renal inflammatory infiltration and kidney intumesce were alleviated in all three SPE groups. CONCLUSIONS: SPE may be a potential therapeutic drug for glomerulonephritis.

Bioequivalence of two oseltamivir formulations in healthy Chinese volunteers
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BACKGROUND: The aim of this study was to compare the bioavailability of a new generic formulation of oseltamivir 75-mg capsule (test) and a branded formulation Tamiflu® (reference) to meet regulatory criteria for marketing the test product in healthy Chinese male volunteers. METHODS: This single-dose, randomized-sequence, open-label, two-period crossover study was conducted in fasted healthy Chinese male volunteers, who first received a single oral dose of the test or reference formulation with a 7-day washout period, and then the alternative formulation. The study drug was administered after a 10-hour overnight fast. Blood samples were collected at baseline and at 0.25, 0.5, 0.75, 1.0, 1.5, 2, 4, 6, 8, 10, 12, 24, and 36 hours after administration of the study drug. Plasma concentrations of the parent oseltamivir and its metabolite oseltamivir carboxylate were determined using an LC-MS/MS method. The formulations were considered bioequivalent if the 90% confidence intervals (CIs) for the log-transformed values were within the predetermined equivalence range (70 - 143% for Cmax, 80 - 125% for AUC) according to the guidelines of the State Food and Drug Administration of China. Adverse events (AEs) were monitored throughout the study based on clinical parameters and patient reports. RESULTS: Characteristics of the 20 male volunteers included were as follows: mean age 23 (± 0.7, SD) years (range 21 - 24 years); weight 69 (± 7.1) kg (range 60 - 88 kg); height 177 (± 5.9) cm (range 168 - 192 cm). All included subjects completed the study. The mean geometric ratio between the test and reference formulations of oseltamivir was 99.5% (90% CI), 86.3 - 114.8%) for Cmax, 104.4% (95.7 - 113.9%) for AUC0-t, and 104.4% (95.6 - 113.9%) for AUC0-∞. That of oseltamivir carboxylate was 103.7% (90% CI, 95.3 - 112.8%) for Cmax, 101.7% (96.6 - 107.1%) for AUC0-t, and 101.4% (96.5 - 106.5%) for AUC0-∞. There was no significant difference in pharmacokinetic parameters between the two groups. Only 1 AE (nausea) occurred in 1 subject who received the test formulation; the AE resolved without any treatment. CONCLUSIONS: The result of this single-dose study indicated that the test formulation of oseltamivir capsule met the Chinese regulatory criteria for bioequivalence vs. the reference formulation in fasted healthy Chinese male volunteers.
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LC-MS/MS analysis and pharmacokinetic study on five bioactive constituents of Tanreqing injection in rats.

Tanreqing injection (TRQ), a well-known traditional Chinese medicine formula, is commonly used to treat respiratory diseases. In the present study, a rapid, selective, and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated to simultaneously determinate the plasma contents of 5 major constituents of TRQ, including chlorogenic acid (CHA), caffeic acid (CFA), baicalin (BA), ursodeoxycholic acid (UDCA) and chenodeoxycholic acid (CDCA) in rats after intravenous administration of TRQ. Chromatographic separation was performed on an Agilent Zorbax SB-C18 column (3.5 µm, 100 mm × 2.1 mm), with acetonitrile and 0.1% aqueous formic acid as mobile phase at a flow rate of 0.3 mL·min-1. The calibration curves were linear over the ranges of 27.0-13 333.0 ng·mL-1 for CFA, 30.0-14 933.0 ng·mL-1 for CHA, 50.0-50 333.0 ng·mL-1 for BA, 550.0-55 000.0 ng·mL-1 for UDCA, and 480.0-48 000.0 ng·mL-1 for CDCA, respectively. Intra- and inter-day precisions (relative standard deviations, RSDs) were from 3.11% to 14.08%. The extraction recoveries were greater than 71% and accuracy (relative recovery) was from 89% to 137% for all analytes, except endogenous bile acids. This validated method was successfully applied to the first pharmacokinetic study of CFA, CHA, BA, UDCA and CDCA in rat plasma after intravenous administration of TRQ.

Metabolite profiling of Zi-Shen pill in rat biological specimens by UPLC-Q-TOF/MS.

This study aimed to profile the chemical constituents of Zi-Shen pill (ZSP) and its metabolites in plasma, urine, and prostate tissue, after administration into rats. Based on the chromatographic retention behavior, fragmentation patterns of chemical components, published literatures, and literature databases, an UPLC-Q-TOF/MS (LC-TOF/MS) method was established to identify the components of ZSP and its metabolites in biological samples. A total of 101 compounds were identified and tentatively characterized from the ZSP, including alkaloids, xanthones, and timosaponins. Except for 33 prototype components, 22 metabolites were detected in the plasma, urine, and prostate, and mainly came from Phellodendri Amurensis Cortex and Anemarrhenae Rhizoma. It was found that glucuronidation and sulfation were the major metabolic processes of xanthones, while oxidation, demethylation, and glucuronidation were the major metabolic pathways of alkaloids. In summary, the present study provided important chemical information on the metabolism of ZSP, indicating that alkaloids might be able to be absorbed into the prostate. The results provided a basis for further studies of the mechanisms of action for ZSP.

Fingerprint analysis of Zhimu-Huangbai herb pair and simultaneous determination of its alkaloids, xanthone glycosides and steroidal saponins by HPLC-DAD-ELSD.

AIM: To develop and validate a high performance liquid chromatography (HPLC) coupled with diode array and evaporative light scattering detectors (DAD-ELSD) method for the quantitative determination and fingerprint analysis of ten active constituents in three chemical classes (namely, xanthone glycosides, steroidal saponins, and alkaloids) in Zhimu-Huangbai herb pair (ZB). METHOD: Chromatographic separation was performed on a Diamonsil C18 column (4.6 mm × 250 mm, 5 µm, Dikma) by gradient elution using acetic acid in acetonitrile solution at a flow rate of 1.0 mL·min(-1) at 260 nm. The drift tube temperature of ELSD was set to 60 °C and nebulizer gas pressure was 4.0 Bar. Method validation was performed to assure its linearity, limits of detection and quantification, precision, repeatability, stability, and accuracy. RESULTS: The HPLC-DAD-ELSD method allowed the quantification of ten compounds (phellodendrine, jatrorrhizine, palmatine, berberine, neomangiferin, mangiferin, timosaponin E-I, timosaponin B-II, timosaponin B, and timosaponin A-III), and was successfully applied to fingerprint analysis for ten batches of ZB samples. CONCLUSION: This was the first time to apply the combination of DAD and ELSD for the simultaneous determination of ten active ingredients in ZB. The results showed that the combination of quantitative analysis for marker ingredients and chemical fingerprint for the TCM herb pair provides a potentially powerful, widely introduced, and internationally accepted strategy for assessment of complex TCM formulas.

An HPLC-MS/MS method for the quantitative determination of platycodin D in rat plasma and its application to the pharmacokinetics of Platycodi Radix extract.

AIMS: To develop an HPLC-MS/MS method for the quantification of platycodin D (PD) in rat plasma, and to acquire the main pharmacokinetic parameters of PD after oral administration of pure PD or of Platycodi Radix extract (PRE) containing PD. METHOD: Plasma samples were pretreated with solid-phase extraction using an Oasis® HLB SPE cartridge. Madecassoside was used as the internal standard (IS). Chromatographic separation was achieved on an ODS column (100 mm × 2.1 mm i.d., 3.5 µm) with a mobile phase consisting of acetonitrile/water (30 : 70, V/V) containing 0.1 mmol·L(-1) ammonium acetate at a flow rate of 0.25 mL·min(-1). The detection was performed on a triple quadruple tandem mass spectrometer using an electrospray ionization (ESI) source with a chromatographic run time of 3.0 min. The detection was operated by multiple reaction monitoring (MRM) of the transitions of m/z 1 223.6→469.2 for PD and of m/z 973.6→469.2 for madecassoside (IS), respectively. RESULTS: The calibration curve was linear from 5 to 2 000 ng·mL(-1) (r(2) >0.99) with a lower limit of quantification (LLOQ) of 5 ng·mL(-1). The intra- and inter-day precision (relative standard deviation, RSD) values were below 15% and the accuracy (relative error, RE) was from -15% to +15% at three quality control (QC) levels. Plasma concentrations of PD were determined for 24 h after i.v. administration of PD, and oral administration of PD and PRE, respectively. The absolute oral bioavailability of PD in rats was found to be (0.48 ± 0.19)% when administered PD, and to be (1.81 ± 0.89)% when administered PRE. CONCLUSION: The developed HPLC-MS/MS method was successfully applied to assess the pharmacokinetic parameters and oral bioavailability of PD in rats after administration of PD and Platycodi Radix extract.

(2RS)-2-(2,4-Difluoro-phen-yl)-1-[(4-iodo-benz-yl)(meth-yl)amino]-3-(1H-1,2,4-tri-azol-1-yl)propan-2-ol.

IN THE TITLE COMPOUND (COMMON NAME: iodiconazole), C(19)H(19)F(2)IN(4)O, there is an intra-molecular O-H⋯N hydrogen bond and mol-ecules are linked by weak inter-actions only, namely C-H⋯N, C-H⋯O and C-H⋯F hydrogen bonds, and π-electron ring-π-electron ring inter-actions between the triazole rings with centroid-centroid distances of 3.725 (3) Å.

Determination of bioactive compounds in Cortex Phellodendri by high-performance liquid chromatography.

An HPLC method combined with a photodiode array detector was developed for quantitative determination of five bioactive compounds that belong to two subclasses, including limonin, phellodendrine, jatrorrhizine, palmatine, and berberine in Cortex Phellodendri. The analysis was performed on an Agilent Diamonsil C18 column (4.6 x 250 mm, 5 microm) using a gradient of acetonitrile and 0.3% aqueous diethylamine phosphate (v/v), a flow rate of 0.8 mL/min, and a detection wavelength of 220 nm. The calibration curve was linear over the range of 2.5-100.0 microg/mL for both phellodendrine and jatrorrhizine, 5.0-200.0 microg/mL for palmatine, and 7.5-300.0 microg/mL for both berberine and limonin. The average recoveries ranged from 97.56 to 102.53% with RSD < or = 1.00%. Samples from different geographical locations were analyzed to evaluate the applicability of the established method, and the results indicated that the method was efficient, sensitive, and reliable for determining limonin and four alkaloids in Cortex Phellodendri.