Study Of Cyclovirobuxine D Controlled-Release Tablets、Pharmacokinetics And Pharmacodynamics Of Its PEG Derivatives

Cyclovirobuxine D,Huangyangning,is an alkaloid extracted from Buxus microphylla Sieb.et Zucc.Var.Sinica Rehd.et wils and other plants of the same species.It has been proved with its clinical application in treating coronary heart disease and arrhythmia,and it has been recorded by ChP,2005 Vol.â… .This study laid emphasis on of cyclovirobuxine D osmotic pump controlled release tablets（Huangyangning controlled-release tablet）.Established cyclovirobuxine D controlled-release tablets quality standards.Studied pharmacokinetics ofcyclovirobuxine D controlled-release tablets and cyclovirobuxine D tablets（Huangyangning tablet） in dogs and pharmacokinetics of tablets in healthy volunteers.Studied pharmacodynamics of PEG derivatives of cyclovirobuxine D initially.The main contents read as follows:1.Study of cyclovirobuxine D osmotic pump controlled-release tabletsCyclovirobuxine D was selected as the model drug to prepare cyclovirobuxine D osmotic pump controlled-release tablets.In formulation design,We choiced sodium chloride and mannitol as osmotic pressure active substances,tartaric acid as cosolvent, PEO as promoting agent,to PEG1500 as pore-foaming.Cumulative release F₂₁ of R² and 21h investigation index and used acid dyes though spectrophotometry to determine their release.The formulation optimize tablets core according to orthogonal design.On the basis of determining tablets core,further studied drug release on the coating weight,aperture.Through the study and screening of formulation and technique,we come to a better formulation:tartaric acid 5 mg,PEO1mg,sodium chloride/mannitol（1:1）,PEG15%,coating weight gain of 4.5%-5.5%,orifice size 0.4-0.6 mm.2.Established cyclovirobuxine D osmotic pump controlled-release tablets quality standards Established cyclovirobuxine D quality standards in order to a comprehensive control quality of cyclovirobuxine D controlled-release tablets from all perspective.Established color reaction and TLC of cyclovirobuxine D.The content determination was established for the controlled-release tablets of cyclovirobuxine D by ultraviolet spectrophotometry.The validated method was studied and established release limit indicators of controlled-release tablets of cyclovirobuxine D.According to optimized formulation,prepare the three batches cyclovirobuxine D controlled-release tablets.Their quality were preliminary investigated.It results show that the three batches controlled-release tablets in 21h showed a good zero release and other basically indicators were the same.3.Pharmacokinetics of the cyclovirobuxine D controlled-release tablets in dogsLC-ESI-MS method for determination cyclovirobuxine D in the dogs plasma was developed.Cyclovirobuxine D and the internal standard were extracted from the dogs plasma using liquid-liquid extraction,then separated on a MetaChem Nucleosil CN column.The mobile phase consisted of methanol-acetate buffer（90:10）.Detection is performed on a HPLC-MS equipped with ESI and operated in positive ionization mode.Cyclovirobuxine D quantity was realized by computing the peak area ratio（cyclovirobuxine D m/z 403.5,thebain m/z 312）.The linear calibration curve was obtained in the concentration range of 0.5-200ng/ml.The low limit of quantitative was 0.2ng/ml.The RSD of within-days and between-day were less than 15%.The validatedmethod is suitable for quantitative determination of cyclovirobuxine D in dogs plasma in pharmacokinetics study.A dose of 50mg each of cyclovirobuxine D orally Huangyangning controlledrelease tablets and Huangyangning tablets was given to six dogs in a randomized crossover study.The concentration of cyclovirobuxine D in plasma was determined by LC-ESI-MS method.After a single dose,the pharmacokinetic parameters for orally Huangyangning tables and reference were as follows:Cmax,（25.41±3.53） and （43.35±4.55）ng/ml;Tmax,（12.00±8.05） and（4.67±1.37）h;AUC_0-Ï„,（419.94±17.80）and （408.31±14.4）h·pg/ml.AUC_0-∞,（452.44±26.89）and（434.7±110.49） ng·h/ml.Variance analys is and thetwo one-sided test for Cmax and AUC_0-∞ showed Cmax decreased more than tables.AUC_0-∞ was no significant difference.Tmax had oblivious difference. Tmax decrease obliviously for Huangyangning controlled-release tablets.A multiple dose modes of each of cyclovirobuxine D orally Huangyangning controlled-release tablets（once everyday,50mg once） and Huangyangning tablets （twice everyday,25mg once）was given to six dogs in a randomized crossover.After sevens days orally given medicine,cyclovirobuxine D plasma concentration reached steady state level in the fifth day.The relative bioavailability of Huangyangning controlled release tablets was 97.43%±5.19%.After a multiple dose,the pharmacokinetic parameters for Huangyangning tables and reference were as follows:Cmax,（59.36±5.58）and（64.70±7.17）ng/ml.Tmax,（13.00±1.79）and（3.67±0.52）h,Cmin,（20.72±3.23）and （20.25±4.39）ng/ml,Cav,（36.83±1.61）and（37.85±1.56）ng/ml,AUCss/D,（17.68±0.77）and （18.71±0.75）h/ml,fluctuation degree（DF） was（1.04±0.20）and（1.17±0.21）.Variance analysis and two one-sided test for Cmax and AUCss/D showed no difference in two preparation.Non-parametric test for Tmax had oblivious difference.Tmax decrease obliviously for Huangyangning controlled-release tablets.The results showed that plasma concentration of Huangyangning controlled-rel-ease tablets,compared with Huangyangning tablets,increased slowly released a longer time.Peak time was delayed,and it has a good controlled release characteristics. Both are bioequivalent.4.Pharmacokinetics of cyclovirobuxine D in healthy volunteersAn SPE-LC-MS/MS method for the determination of cyclovirobuxine D in human plasma was performed.The chromatographic separation was performed on Lichrospher CN column with a mobile phase of 90%methanol containing 0.2%formic acid and 2.5mmol/l ammonium acetate.Cyclovirobuxine D quantity was realized by computing the peak area ratio（cyclovirobuxine D m/z 403.20→372.30,citalopram m/z 325.10→262.10）.The plasma sample were pretreated on a Phenomenex Strata^TM-X 8 B-S100-TAK SPE column.The pharmacokinetic parameters for Huangyangning tables orally were as follows:Cmax,（216.33±139.80）pg/ml;Tmax,（5.70±1.40）h,T_1/2,（54.35±12.00）h;MRT_0-144,（45.19±0.20）h,AUC_0-144（5686.78±1789.47）h·pg/ml;AUC_0-∞,（7404.02±2566.60）h·pg/ml.Medicine was given in ten healthy volunteers.After ten days orally given medicine, plasma concentration of cyclovirobuxine D reached steady-state level in the seventh day.After steady-state level,the pharmacokinetic parameters for Huangyangning controlled-release tablets were as follows:Tmax,（4.60±0.52）h,AUC_0-Ï„,（29572.51±5642.41）h·pg/ml;AUCss,（2433.96±391.26h）pg/ml;Cav,（304.25±48.91）pg/ml;DF, 0.26±0.18. 5.Pharmacodynamics of PEG derivatives of cyclovirobuxine DAcute cerebral ischemia model was established by ligating the common carotid arteies in mice.The living time of mice was observed after given PEG derivatives of cyclovirobuxine D and cyclovirobuxine D.Ruselt showed cyclovirobuxine D has obvious protect effect for acute cerebral ischemia in mice.But PEG derivatives of cyclovirobuxine D had no obvious protect effect.Focal cerebral ischemia and focal cerebral ischema-reperfusion of MACO model was established by ligustrazine.Focal cerebral infarct areas was measured after PEG derivatives of cyclovirobuxine D and cyclovirobuxine D in rats.Rustle showed cyclovirobuxine D could obvious decreased focal cerebral ischema-reperfusion infarct areas.But cyclovirobuxine D had no significant effect for focal cerebral ischema. PEG derivatives of cyclovirobuxine D wasn’t decrease infarct areas of cerebral ischema and cerebral ischema-reperfusion model in rats.In the experiment of hypoxia method of mice under the normal pressure,cyclovirobuxine D and PEG derivatives of cyclovirobuxine D could prolong obviously the living time of mice under the normal pressure and hypoxia.Every batches of PEG derivatives of cyclovirobuxine D showed dose-effect relationship.PEG derivatives of cyclovirobuxine D had more better than cyclovirobuxine D.But both were no significant different.The innovation lies in this paper:1.Carried out cyclovirobuxine D osmotic pump controlled release tablets study firstly,and established its quality standards.2.Carried out comparative pharmacokinetics studies for cyclovirobuxine D osmotic pump controlled-release tablets and tablets in dogs after single dose and multiple dose firstly.It provided a theoretical basis for clinical application in the future.3.Carried out pharmacokinetic study of cyclovirobuxine D tablets in human after a multiple-dose firstly,enables us to the deeper understanding for Cyclovirobuxine Dtablets.New drug development of cyclovirobuxine D provide more experimental data.4.Carried out pharmacodynamic study for the PEG derivatives of cyelovirobuxine D firstly.New drugs research of Cyclovirobuxine D provide a new research idea for the future.