Polyynes, such as facarindiol (FAD) and oplopandiol (OPD), are responsible for anticancer activities of Oplopanax elatus (O. elatus). A novel approach to pharmacokinetics determination of the two natural polyynes in rats was developed and validated using a liquid chromatography-electrospray ionization-mass spectrometry (LC-MS) method. Biosamples were prepared by liquid-liquid extraction using ethyl acetate/n-hexane (V : V = 9 : 1) and the analytes were eluted on an Agilent ZORBAX Eclipse Plus C18 threaded column (4.6 mm × 50 mm, 1.8 μm) with the mobile phase of acetonitrile-0.1% aqueous formic acid at a flow-rate of 0.5 mL·min(-1) within a total run time of 11 min. All analytes were simultaneously monitored in a single-quadrupole mass spectrometer in the selected ion monitoring (SIM) mode using electrospray source in positive mode. The method was demonstrated to be rapid, sensitive, and reliable, and it was successfully applied to the pharmacokinetic studies of the two polyynes in rat plasma after oral administration of polyynes extract of O. elatus.
Administration, Oral ; Animals ; Chromatography, High Pressure Liquid ; methods ; Diynes ; administration & dosage ; pharmacokinetics ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Fatty Alcohols ; administration & dosage ; pharmacokinetics ; Male ; Naphthols ; administration & dosage ; pharmacokinetics ; Oplopanax ; chemistry ; Polyynes ; administration & dosage ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Spectrometry, Mass, Electrospray Ionization ; methods

Our previous report showed that polyacetylene compound, 1-Heptadecene-11, 13-diyne-8, 9, 10-triol (PA) from the root of Cirsium japonicum var. ussuriense has anti-inflammatory activity. In this study we investigated the role of the PA as inhibitor of caspase-1, which converts prointerleukin-1beta (proIL-1beta) to active IL-1beta and is activated by inflammasome involved in the inflammatory process. We tested the effect of PA on the production of pro-inflammatory cytokines, IL-1beta in murine macrophage cell line, RAW264.7. PA inhibited lipopolysaccharide (LPS)-induced IL-1beta production by macrophages at a dose dependent manner. PA also suppressed the activation of caspase-1. The mRNA level of ASC (apoptosis-associated spec-like protein containing a CARD), an important adaptor protein of inflammasome, was decreased in the PA treated group. Therefore our results suggest that the anti-inflammatory effect of PA is due to inhibit the caspase-1 activation.
Cell Line ; Cirsium ; Cytokines ; Macrophages ; Polyacetylenes ; RNA, Messenger

OBJECTIVETo develop a green and rapid method for extraction of lobetyolin from C. pilosula.

METHODExtraction of lobetyolin from C. pilosula with supercritical carbon dioxide in the presence of ethanol was studied. The effects of pressure, temperature, volume of cosolvent and extraction time on efficiency and their interactive relationships were discussed, based on central composite design and response surface methodology (RSM).

RESULTThe key effect factor was volume of cosolvent. The extraction yield of lobetyolin was 0.078 6 mg x g(-1) when C. pilosula (40-60 mesh) was extracted at 30 MPa, 60 degrees C and 2 L x min(-1) (as CO2 in normal pressure and temperature) for 100 minutes with supercritical CO2 and 1 mL x min(-1) ethanol as dynamic cosolvent.

CONCLUSIONThis result is better than that obtained from traditional method. Therefore, the optimized process is valuable for extraction of lobetyolin from C. pilosula.

Carbon Dioxide ; chemistry ; Chemical Fractionation ; methods ; Codonopsis ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Ethanol ; chemistry ; Polyacetylenes ; chemistry

Here, we designed to examine the anti-inflammatory effects on RAW264.7 cells and the immunosuppressive effects by evaluating interleukin-2 (IL-2) production in Jurkat T cells using a MeOH extract of Panax notoginseng roots. The results showed that the MeOH extract inhibited the synthesis of nitric oxide (NO) in a dose-dependent manner (IC₅₀ value of 7.08 µg/mL) and displayed effects on T cell activation at a concentration of 400 µg/mL. In efforts to identify the potent compounds, bioactivity-guided fractionation of the MeOH extract and chemical investigation of its active CH₂Cl₂-, EtOAc-, and butanol-soluble fractions led to the successful isolation and identification of eleven compounds, including two polyacetylenes (1, 2), a steroid saponin (3), seven dammarane-type ginsenosides (4 – 10), and an oleanane-type ginsenoside (11). Among them, compound 11 was isolated from this plant for the first time. Compound 2 exhibited potent inhibitory effects on NO synthesis and an immunosuppressive effect with IC₅₀ values of 2.28 and 65.57 µM, respectively.
Ginsenosides ; Interleukin-2 ; Nitric Oxide ; Panax notoginseng ; Panax ; Plants ; Polyacetylenes ; Saponins ; T-Lymphocytes

OBJECTIVETo determine lobetyolin in the root of Codonopsis tangshen from the various cultivation areas.

METHODA Supelco Discovery C18 Column (4.6 mm x 250 mm, 5 microm) was used with acetonitrile-0.5% acetic acid in water (20:80) as the mobile phase and UV detection was at 268 nm.

RESULTTwenty-four batches of the samples were analyzed. The content of lobetyolin ranged from 0.0403-0.9667 mg x g(-1).

CONCLUSIONThe method was simple, reproducible and reliable. It can be used to control the quality of C. tangshen.

China ; Chromatography, High Pressure Liquid ; methods ; Codonopsis ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Plant Roots ; chemistry ; Polyacetylenes ; analysis ; Quality Control

OBJECTIVETo develop an HPLC method for determination of two polyacetylenes, lobetyolin and lobetyolinin, in Herba Lobeliae Chinensis.

METHODC18 column was used with the mobile phase consisted of acetonitrile and water. Linear gradient elution from 10% to 40% acetonitrile in 25 min was applied, at the flow rate of 1.0 mL x min(-1), the detection wavelength was at 267 nm.

RESULTLower contents of lobetyolin and lobtyolinin were found in collected samples of Herba Lobeliae Chinensis. The highest amounts of lobetyolin and lobetyolinin were found to be 0.461 and 0.436 mg x g(-1) in a sample procured from Hong Kong. However, there were no lobetyolin and lobetyolinin in some of the samples.

CONCLUSIONA simple and effective HPLC method to analyze the two polyacetylenes in Herba Lobeliae Chinensis was established. It could be applied for the quality control of this herb.

Chromatography, High Pressure Liquid ; methods ; Lobelia ; chemistry ; Molecular Structure ; Plants, Medicinal ; chemistry ; Polyacetylenes ; analysis ; chemistry ; Quality Control ; Reproducibility of Results

AIMTo compare two methods, the total radioactivity assay (RA method) and the radioactivity assay after separation with high performance liquid chromatography (HPLC-RA method).

METHODS125I-Lidamycin was prepared by Iodogen method and separated by size exclusive high performance liquid chromatography. The pharmacokinetic parameters of lidamycin were assayed by two methods after intravenous injection to mice at the dose of 100 microg x kg(-1), and compared by statistical analysis.

RESULTSThe pharmacokinetic parameters (Vd, T1/2alpha, T1/2beta, K21, K10, K12, AUC and CL) showed significant difference between the two methods (P < 0.05).

CONCLUSIONThe HPLC-RA method was better than the RA method to determine unchanged 125I-lidamycin.

Aminoglycosides ; blood ; pharmacokinetics ; Animals ; Antibiotics, Antineoplastic ; blood ; pharmacokinetics ; Area Under Curve ; Chromatography, High Pressure Liquid ; methods ; Enediynes ; Female ; Iodine Radioisotopes ; Isotope Labeling ; Male ; Mice ; Sensitivity and Specificity

AIMTo investigate the chemosensitivity to lidamycin (C-1027) in mdr1 gene overexpressing cancer cell lines established by drug induction and by gene-transfection.

METHODSDNA was cloned by RT-PCR and then eukaryotic expressing recombinant plasmid pcDNA3. 1/mdrl was constructed. Using Lipofectamine 2000, a strain of stably transfected human hepatoma cancer cells, HepG2/mdrl, was obtained. The mdr1 mRNA level, P-glycoprotein (P-gp) level and the activity of P-gp to extrude drugs in cancer cells were determined by RT-PCR, immunofluorescence analysis and rhodamine 123 efflux assay. The chemosensitivity of cancer cells with low or high mdr1 expression to lidamycin and other antitumor drugs was tested by MTT assay.

RESULTSThe mdr1 mRNA and P-gp levels in KBv200, MCF-7/ADR, and stably transfected HepG2/mdr1 cells were much higher than that in respective parent KB, MCF-7 and HepG2 cells. The IC50 values of lidamycin for KBv200, MCF-7/ADR and HepG2/mdrl cells were (0.24 +/- 0.20) nmol x L(-1), (0.028 +/- 0.011) nmol x L(-1), and (0.020 +/- 0.011) nmol x L(-1), respectively. Compared with parental cells, the values of resistant fold for KBv200, MCF-7/ADR and HepG2/mdr1 cells to lidamycin were 6.8, 1.6 and 1.3 fold; to adriamycin were 37.2, 181.3 and 8.8 fold; to taxol were 336.8, 49.2 and 40.3 fold, respectively.

CONCLUSIONLidamycin is highly active to multidrug resistant cancer cells. The chemosensitivity of those resistant cancer cells to lidamycin is approximately at the similar level as that of parent cancer cells.

ATP-Binding Cassette, Sub-Family B, Member 1 ; analysis ; genetics ; Aminoglycosides ; pharmacology ; Antibiotics, Antineoplastic ; pharmacology ; Cell Line, Tumor ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Enediynes ; pharmacology ; Genes, MDR ; Humans ; Neoplasms ; drug therapy ; pathology ; Transfection

This paper is to report the study of the metabolism of lidamycin in vitro including in plasma and microsomes to guide clinical therapy. Lidamycin was quantified by detecting its active ingredient using HPLC-MS/MS. The metabolic stability of lidamycin in rat, Beagle dog, monkey and human plasma and liver microsomes, and its inhibition to cytochrome P450 isoforms in human liver microsomes were studied. Results showed that lidamycin was metabolized in the four species of plasma, and the sequence of metabolic rates in plasma were in rat > in dog > in human > in monkey. But among the four species of liver microsomes, lidamycin was metabolized only in monkey liver microsomes. There was almost no inhibition to cytochrome P450 isoforms at the concentrations of between 0.0005 and 10 ng x mL(-1). Therefore, the property of lidamycin metabolism in human is similar with that in dog, and metabolism of other drugs would not be decreased by cytochrome P450 as used along with lidamycin in clinic.
Aminoglycosides ; blood ; metabolism ; Animals ; Antibiotics, Antineoplastic ; blood ; metabolism ; Chromatography, High Pressure Liquid ; Cytochrome P-450 CYP1A2 ; metabolism ; Cytochrome P-450 Enzyme System ; metabolism ; Dogs ; Enediynes ; blood ; metabolism ; Enzyme Activation ; Humans ; Macaca ; Microsomes, Liver ; metabolism ; Rats ; Tandem Mass Spectrometry

AIMA bioassay method was established for the determination of active concentrations of lidamycin and studied its pharmacokinetics in mice and dogs.

METHODSCytotoxicity of lidamycin in vitro was used to determine drug serum concentrations in vivo.

RESULTSValidity of methodology met the requirements of pharmacokinetic study. The concentration-time profile in mice after iv lidamycin of 100, 50 and 10 microg x kg(-1) was best fitted with 2-compartmental model with T1/2alpha and T1/2beta of 0.77-1.8 min and 5.6-7.2 min, respectively. The AUC were 2851.3, 887.8 and 166.4 microg x min x L(-1), respectively and increased with dose nonlinearly. There were similar trends between AUC and the potency of tumor growth inhibition. After iv lidamycin of 12 microg x kg(-1) in dogs, the concentrations of lidamycin decreased rapidly and the AUC was 16 microg x min x L(-1), which were lower and quicker than those in mice. The levels in serum after second administration at day 15, were lower than those of the first.

CONCLUSIONActive concentrations and pharmacokinetics of lidamycin were obtained by bioassay method successfully. There are species differences and single and multi-dosing differences in the pharmacokinetics of lidamycin.

Aminoglycosides ; blood ; pharmacokinetics ; pharmacology ; Animals ; Antibiotics, Antineoplastic ; blood ; pharmacokinetics ; pharmacology ; Area Under Curve ; Biological Assay ; Dogs ; Enediynes ; Female ; Humans ; Injections, Intravenous ; KB Cells ; metabolism ; Liver Neoplasms ; pathology ; Male ; Mice ; Neoplasm Transplantation ; Sarcoma 180 ; pathology ; Species Specificity