AIMTo establish a RP-HPLC method for determination of cyclovirobuxine D.

METHODSCyclovirobuxine D reacted with a derivative reagent 1-naphthyl isocyanate in chloroform to form fluorescence derivatives, stopped the reaction by adding the mobile phase and then directly injected the solution into the chromatograph to seperate it by RP-HPLC. The analysis was carried out on C18 column, the mobile phase is methanol-water (85:15), the excitation wavelength was set at 305 nm, emission at wavelength 385 nm, and the flow rate was 1 mL.min-1. The effect of several factors including the reaction medium, temperature, time and amount of 1-naphthyl isocyanate on the yield of the derivatization was also investigated systematically.

RESULTSA simple and rapid RP-HPLC method for the simultaneous isolation and analysis of cyclovirobuxine D and its related substances was developed, and the absence of interference between the derivative peak responses of cyclovirobuxine D and its related substances were verified by UV diode array detecter and MS. The linearity was obtained from 0.75 microgram.mL-1 to 2.5 micrograms.mL-1 of cyclovirobuxine D derivatives with a correlation coefficient of 0.9991. The detection limit of cyclovirobuxine D derivative was 1 ng.mL-1, the repeatability of derivatization was good with relative standard derivation no more than 1.2% and derivative was stable within 48 h. The method described conforms to the validation of China Pharmacopiea compendial methods used for pharmaceutical products in general.

CONCLUSIONThe established method is proved to be reliable quantitative method for the quality control of cyclovirobuxine D.

Buxus ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Plants, Medicinal ; chemistry ; Quality Control

OBJECTIVETo establish a new method for the determination of cyclovirobuxine-D in huangyangning tablets.

METHODAgilent Zorbax Extend C18 (4.6 mm x 250 mm, 5 microm) column with temperature 40 degrees C was used. The mobile phase was 0.3% diethylamine methanol solution and 0.3% diethylamine water solution, with a linear gradient of 0.3% diethylamine methanol solution from 78% to 95% in 9 minutes, and then maintained for 6 minutes. The flow rate was 0.8 mL x min(-1). Evaporative light scattering detector was used.

RESULTThe calibration curve was linear at a range of 0.57-11.44 microg for the cyclovirobuxine D (r = 0.9996). The average recovery was 97.2% and RSD was 1.1% (n = 9).

CONCLUSIONThis method is rapid, simple, and reproducible, and can be used as a quality control method for this preparation.

Buxus ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; administration & dosage ; analysis ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Tablets

AIMTo search for compounds through structural modification of cyclovirobuxine D, using 20 or 21-aminosteroids as lead compound for the treatment of cerebrovascular diseases with better lipid peroxidation inhibitory activity.

METHODSAccording to rational drug design principle, a series of cyclovirobuxine D analogues were prepared, and their bioactivities were tested.

RESULTSFour new compounds were obtained and confirmed by spectra.

CONCLUSIONLipid peroxidation inhibitory effects of cyclovirobuxine D analogues were tested by using TBA method. Some compounds showed better activity than that of cyclovirobuxine D.

Animals ; Buxus ; chemistry ; Drugs, Chinese Herbal ; chemical synthesis ; isolation & purification ; pharmacology ; Lipid Peroxidation ; drug effects ; Malondialdehyde ; metabolism ; Molecular Structure ; PC12 Cells ; Plants, Medicinal ; chemistry ; Rats

AIMTo determine the effects of cyclovirobuxine D (CD) on intracellular Ca2+ mobilization and L-type Ca2+ current (I(Ca-L)) in isolated rat cardiomyocytes.

METHODSThe effects of CD on the amplitude of I(Ca-L) and intracellular Ca2+ mobilization induced by KCl and caffeine were studied with the method of patch-clamp technique and laser scanning confocal microscopy in rat ventricular myocytes.

RESULTSCD decreased the amplitude of I(Ca-L) in a concentration-dependent manner. At 10 mV, 1 and 10 micromol x L(-1) CD decreased I(Ca-L) density from (- 9.9 +/- 1.8) pA/pF to (-6.4 +/- 1.4) and (-4.2 +/- 0.6) pA/pF, respectively. Confocal experiments showed that intracellular fluorescent intensity (FI) value of [Ca2+] in control resting level was not changed by 1 and 10 micromol x L(-1) CD. [Ca2+] increase in response to KCl could not be reduced by CD. The rise of [Ca2+]i in response to caffeine was further enhanced by pretreatment with CD.

CONCLUSIONCD decreased I(Ca,L) in a concentration-dependent manner and increased [Ca2+]i release induced by caffeine in rat ventricular cardiomyocytes.

Animals ; Buxus ; chemistry ; Calcium ; metabolism ; Calcium Channels, L-Type ; drug effects ; Cell Separation ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Heart Ventricles ; cytology ; Male ; Myocytes, Cardiac ; metabolism ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar

AIMTo search for new compounds for the treatment of cardiovascular diseases by structural modification of cyclovirobuxine D.

METHODSAccording to rational drug design principle, a series of cyclovirobuxine D analogues were prepared, and their bioactivities were tested.

RESULTSTen new compounds were syntheized and confirmed by spectra.

CONCLUSIONEndurance lacking oxygen activity and antiarrhythmia effects of some analogues of cyclovirobuxine D were tested. Some compounds showed better activity than cyclovirobuxine D.

Anaerobic Threshold ; drug effects ; Animals ; Anti-Arrhythmia Agents ; chemical synthesis ; pharmacology ; Buxus ; chemistry ; Chloroform ; Drugs, Chinese Herbal ; chemical synthesis ; isolation & purification ; pharmacology ; Female ; Male ; Mice ; Molecular Structure ; Plants, Medicinal ; chemistry ; Random Allocation ; Ventricular Fibrillation ; chemically induced ; prevention & control

AIMTo search for compounds for the treatment of cardiovascular diseases through prodrug structural modifications of cyclovirobuxine D, a single efficient composition distilled from Box plant in China, which was used to treat angina and myocardial infarction.

METHODSAccording to prodrug design principle, a series of cyclovirobuxine D analogues were prepared, suc as succinate, phosphate and amino acid ester, and their biological activities were tested.

RESULTSSeven new compounds were obtained and confirmed with 1H NMR, MS, and element analysis.

CONCLUSIONIn pharmacology experiment, for treating arrhythmia induced by aconitine, succinate and amino acid ester of cyclovirobuxine D (I and VII) showed better activities than that of cyclovirobuxine D. The normal rhythm of the heart duration of I and VII were ( 11.53 +/- 7.62) min and (12.68 +/- 9.25) min, compared with 0.9% NaCl solution and cyclovirobuxine D, (2.36 +/- 1.68) min and (10.25 +/- 6.59) min (P < 0.01), respectively. Another pharmacology experiment, for treating arrhythmia induced by chloroform, the negative ratio of I and VII were 80% and 82%, compared with 0.9% NaCl solution and cyclovirobuxine D, 43% and 52% (P < 0.05), respectively. The difference between new compounds and cyclovirobuxine D was distinct.

Aconitine ; Animals ; Anti-Arrhythmia Agents ; chemical synthesis ; pharmacology ; Arrhythmias, Cardiac ; chemically induced ; physiopathology ; Buxus ; chemistry ; Chloroform ; Drugs, Chinese Herbal ; chemical synthesis ; pharmacology ; Female ; Heart Rate ; drug effects ; Male ; Mice ; Plants, Medicinal ; chemistry ; Prodrugs ; chemical synthesis ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the effect of cyclovirobuxinum-D (CVB-D) on cerebral ischemia-reperfusion injury in rats and explore its mechanisms.

METHODOne hundred and twenty rats were randomly divided into three CVB-D groups (2, 1, 0.5 mg x kg(-1)), Nimodipine group (2 mg x kg(-1)), model group and sham operated group, 20 rats each group. Rat cerebral ischemia-reperfusion injury model was induced by middle cerebral artery occlusion, the nerve injury symptoms was evaluated, the level of SOD and MDA in brain tissue were determined, the concentration of intracellar Ca2+ of brain was measured, and the pathological change of brain was also observed.

RESULTCVB-D could improve the nerve injury symptoms, reduce the infarction area of brain, the concentration of intracellar Ca2+ and the level of MDA, increase the activity of SOD, and decrease the pathological change of brain.

CONCLUSIONCVB-D has protective effect on cerebral ischemia-reperfusion injury in rats.

Animals ; Brain ; drug effects ; metabolism ; pathology ; Buxus ; chemistry ; Calcium ; metabolism ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Infarction, Middle Cerebral Artery ; complications ; Male ; Malondialdehyde ; metabolism ; Neuroprotective Agents ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; etiology ; metabolism ; prevention & control ; Superoxide Dismutase ; metabolism