中药复方谱动学与谱效动力学差异性的研究

目的:对中药多成分体系谱动学与谱效动力学的数学模型及参数差异进行研究,回答中药多成分药物动力学参数能否指导中药临床用药问题。方法:在已建立的中药四谱学理论基础上,主要对谱动学与谱效动力学的数学模型及参数的差异进行对比分析,找出其特点与应用条件。结果:从定量药理学的角度,分析了单成分与中药多成分药物动力学,药效动力学的函数表达、零、一、二阶矩的通用式及运算结果,总量药物动力学参数由单个成分的药物动力学参数决定,总量药效动力学参数由单成分的药物动力学参数与药效系数共同决定,当中药各成分的构成比决定了,各成分的效应系数一定时,药效动力学参数与药物动力学参数对应。结论:中药药效决定于药物动力学参数及效应系数。单成分药物动力学参数能独立指导药物临床用药是谱效动力学的特例,不适用于多成分体系,不能用之来解决中药多成分谱效动学的问题。

中药谱动学与谱效动力学关联性研究现状及其关键问题解决的对策探讨

安全、有效和可控是药品基本属性,量-时-效关系的阐明为关键科学和技术问题,长期制约中药复方作用模式的解析和药物创制。单成分的药效动力学(pharmacodynamics,PD)与药动学(pharmacokinetics,PK)呈相关性,可用PK的量-时关系代替PD的效-时关系,而中药复方谱动学(polypharmacokinetics,PPK)参数能否代替谱效动力学(polypharmacodynamics,PPD)参数成为研究的难点。通过生物超分子体按“印迹模板”产生“气析”药效和量-时-效定量函数怎样实现可积分的视角,分析中药PPK与PPD研究方法的不足,探讨中药多成分与单成分在量-时-效关系研究方法学上的差异,阐明PK/PD模型仅适用单成分量-时-效关系表达,而中药的PPK/PPD模型必须单独创立的客观必然性,提出中药PPK与PPD关联的成分簇按“印迹模板”整合简化和对不可积分项用幂级数展开积分的关键问题解决对策,为中药复杂体系解析模式研究提供理论依据。

中药复方网络药理学及其动力学关键技术问题的探讨

目的:从多成分网络药理学及动力学研究现状探讨中药复方网络药理学及其动力学关键技术问题的研究方法。方法:分析中医药学科与网络药理学科研究的特点,根据中药复方药效物质作用的本源,结合中药现代化的要求,提出中药复方网络药理学研究的总体思路与方法。结果:与单成分有效性研究相比,中药复方的网络药理学问题分静态与动态两种形式,静态是指中药复方入口前的原成分群与某一时间点固定的网络关系,如中药复方采用指纹图谱表达,亦为网络谱效学问题;动态的网络药理学问题是指入口后中药动态代谢后血样中成分群与各时间点的网络药效动力学问题,与指纹图谱结合后者为网络谱效动力学,后者包括前者,因此中药复方网络谱效动力学问题是量-比、量-时、量-效三者的有机统一整体;结论:中药复方的网络药理学的关键而又基础的科学问题是基于网络平衡常数表征的网络谱效动力学参数测算问题。

中药制剂单用确定性与合用调配性的关键理论技术问题研究

目的:通过分析中药制剂研究特点,明确该学科的长期发展方向,并提出本学科亟待解决的关键问题及对策。方法:根据中药制剂研究的发展轨迹,结合中医药基础理论、现代新药研究技术与本人从事中医药现代化研究经验,从中药制剂发展方向、特点、亟需解决的关键问题与对策等方面进行探讨。结果:建立以结构与作用明确的多成分群为原料,按中医药基础理论或现代网络医药为指导,以来源方便的动、植、矿物为原料,单用确定性(有效、稳定、可控),合用可调性(可预、最优效、最低毒)整体相统一的单个、群体与中药复方制剂体系。在实现上述目标时,需解决成分有效性归属、遗传稳态性与一次投料量、成分间理化性质迁移规律、提取过程动态性与稳态性规律、制备时整体模型受控与紊湍受控规律、体内外的评价规律、微观质量与宏观质量、单用的确定性与合用的调配性等关键技术问题。结论:中药制剂是体现单用确定性与合用调配性高度统一的整体有效成分群制剂。

Pharmacokinetics of Scutellarin in Dogs

Aim To establish an RP-HPLC method for the determination of scutellarin in plasma and study its pharmacokinetics in dogs. Methods Scutellarin was given to dogs by intravenous injection and determined by RP-HPLC, the mean plasma concentration-time curve was plotted and pharmacokinetic parameters were calculated by program 3p87. Resu;ts The concentration-time curve of scutellarin could be fitted to three-compartment model with T1/2 pi, T1/2 α and T1/2 β being 1.05 ± 0.80 min, 6.99 + 2.76 min and 51.61 + 28.78 min, respectively, Vc being 880.1 + 508.3 mL, CL being 189.6 + 53.8 mL@ min- 1, and AUC0-90 and AUC0-∞ being 574.43 + 133.95 μg@ min@ mL - 1 and 599.34 ± 132.00μg@ min@mL- 1, respectively. Conclusion The fact that the concentrations of scutellarin in plasma declined rapidly after the medication suggested that the T1/2 of scutellarin should be taken into account in drug administration and preparation development.

Pharmacokinetics of Oxiracetam in Healthy Volunteers

Aim To study the pharmacokinetics of oxiracetam after single and multipleintravenous administrations in healthy volunteers. Method A HPLC method was used to determine theserum concentration of oxiracetam after intravenous single dose and daily dose of 2 000 mg for 7 din ten Chinese healthy volunteers. Pharmacokinetic analysis was carried out using Drug And Statisticsoftware. Results The AUC_(0-12), AUC_(0-∞), K_e, t_(1/2), MRT after a single dose of 2 000 mgoxiracetam were 256.26 ± 16.84 μg·mL^(-1)·h, 276.74 ±18.11 μg·mL^(-1)·h, 0.18 ±0.03 h^(-1),3.84±0.64 h, and 4.39 10.39 h, and after multiple doses of oxiracetam were 259.36 ±25.43μg·mL^(-1)·h, 285.59 ±27.38 μg·mL^(-1)·h, 0.17 ±0.04 h^(-1), 4.14 ± 0.82 h, and 4.87 ±0.69 h, respectively. Conclusion The pharmacokinetic parameters of oxiracetam do not differremarkably after single and multiple intravenous administration and there is accumulation in serumafter 2 000 mg multiple intravenous administration once a day fof 7 d.

Pharmacokinetics of Ferulic Acid in Rabbits with BloodStasis

To test and study the Syndrome and Treatment Pharmacokinetics (S & TRK),we studied the pharmacokinetics of ferulic acid in healthy and blood stasis (microcirculation dysfunction)rabbits by RP-HPLC. After a single intravenous injection offenilic acid(5mg/kg)to healthy and blood stasis rabbits, compartment model of ferulic acid serum concentration was fitted and then pharmacokinetic parameters were calculated with a MCPKP program on a COMPAQ 386 compute Important parameters are as follows: In healthy rabbits V_B=0.9525±0.0211 L/kg,V_1=0.2462±0.0381 L/kg, CL_B=1.8133±0.9512 L/h·kg, T_(1/2β)=0.3639±0913, AUC=2.7566±0.8232 μg·h/ml; In blood stasis rabbits V_B=0.7882±0.0321 L/kg,V_1=0.1966±0.0537 L/kg,CL_B=0.8820±0.5481 L/h·kg,T_(1/2β)=0.6193±0.1216 h, AUC=5.6690±2.3541μg·h/ml.Through this experiment we found the sig-nificant differences in the FA's pharmacokinetic parameters between healthy and blood stasis rabbits.The results obtained correspond with S & TPK.

High Resolution Determination of Ondansetron in Human Plasma by HPLC and Pharmacokinetics of Orally Disintegrating Tablets

Ahn To develop a high resolution HPLC method for the determination of ondansetron in human plasma and to study the pharmacokinetics of ondansetron in orally disintegrating tablets. Methods HPLC determination involved liquid-liquid extraction, separation on a CN column and ultraviolet detection at 310 ran with granisetron as an internal standard. Pharmacokinetics and bioequivalence of ondansetron in orally disintegrating tablets by direct compression and conventional 8 mg tablets were evaluated and compared in 20 healthy human male volunteers after a single oral dose in a randomized cross-over study. Results The limit of quantification was 0.25 ng· mL^-1. The recovery was about 85 % or over for ondan setron and about 90% for internal standard. Linearity was good within the concentration range of 0.5 - 50 ng·mL^-1 with r^2 ranging from 0.997 1 to 0.999 9. Intra- and inter-assay coefficients of variation ranged from 1.78% to 2.38% and 3.88% -5.19%, respectively. Accuracies for spiked concentrations of 2.0, 10.0, and 30.0 ng·mL^-1 were 104.7% ±4.4%, 102.2% ± 1.1%, and99.51% ±2.34%, respectively. Pharmacokinetic parameters of AUCo-t, AUCo-∞ , Cmax, Tmax, and T1/2 were 230.2 ± 78.0 ng·h·L^-1 , 265.2± 101.5 ng·h·mL^-1, 35.67 ± 8.94 ng·mL^-l, 1.51 ±0.79 h, and 5.00± 1.41 h for orally disintegrating tablets, respectively. The analysis of variance did not show any significant difference between orally disintegrating tablets and conventional tablets, and 90% confidence intervals fell within the acceptable range for bioequivalence. Conclusion High resolution HPLC method has been set up and applied in pharmacokinetic evaluation of ondansetron in orally disintegrating tablets.

Determination of Sulphonylurea Glimepiride in Dog Serum by RP-HPLC with Pre-column Derivatization

Aim A simple, sensitive and rapid RP HPLC method with pre column derivatization has been developed for the determination of sulphonylurea glimepiride in dog serum. Methods The sulphonylurea glimepiride was extracted from the dog serum using dichlromethane followed by derivatization with DNBF for 20 min at 100℃. The solvent was then evaporated at 60℃ under nitrogen, and the residue was taken up in 100 μL of mobile phase consisting of acetonitrile water (75∶30, v/v). The separation was performed on a Hypersil BDS C18 column with a flow rate of 0 8 mL·min -1 , and the ultraviolet detector wavelength was set at 350 nm. Results Extraction recovery ranged from 75.9% to 83.2%, and methodological recovery was between 96.5% and 109.3%. Within day RSD ranged from 1.5% to 6.3%, and inter day RSD was between 2 9% and 14.8%. The method showed good linearity (R=0.9998). Conclusion The method was simple, convenient and sensitive. The reaction of derivatization was reproducible.

Pharmacokinetics of Active Metabolite of Prulifloxacin in Healthy Chinese

Aim To investigate the pharmacokinetics of NM394 （an active metabolite of prulifloxacin） and evaluate the dose relationship and accumulation characteristics after single and multiple doses of prulifloxacin. Methods Twelve healthy volunteers were given 132.1 mg, 264.2 mg, and 396.3 mg of prulifloxacin tablets in a randomized 3 × 3 crossover design test for single doses trial. With one-week washout period, 264.2 mg of prulifloxacin tablets were given for multiple doses trial. NM394 in plasma was determined by a sensitive HPLC method and its pharmacokinetic parameters were analyzed and evaluated by Drug and Statistics saftware （version 1.0）. Results The Cmax, Tmax, t 1/2, AUC0-24, and AUC0-∞ of NM394 after single doses of 132.1 mg, 264.2 mg, and 396.3 mg of prulifloxacin tablets were 0.64 ± 0.25 μg· mL^-1, 1.06 ± 0.35 μg· mL^-1, and 1.45 ± 0.44 μg· mL^-1 , respectively; Tmax 0.94±0.22 h, 1.02±0.17 h, and 0.98±0.23 h, respectively; t 1/2 8.37±0.70 h, 7.70±0.82 h, and 7.78 ± 0.77 h, respectively; AUC0-24 2.93 ± 0.78 μg· mL^-1· h, 4.39 ± 1.05 μg· mL^-1· h, and 5.55± 1.32 μg·mL^-1 ·h, respectively; AUC0-∞ 3.32±0.84 μg·mL^-1 ·h, 4.82± 1.06 μg·mL^-1 ·h, and 6.10 ± 1.38 μg·mL^-1· h, respectively. And the Cmax, Tmax, t,1/2, AUC0- 24, and AUC0-∞ after multiple doses of 264.4 mg prulifloxacin tablets were 1.20 ± 0.33 μg· mL^- 1, 0.67 ± 0.12 h, 7.38 ± 1.03 h, 5.58 ± 1.25 μg· mL^-1·h, and 6.09 ± 1.24 μg· mL^-1· h, respectively. Conclusion The Cmax and AUC of NM394 are in high correlation with given prulifloxacin doses. There are no differences in pharnacokinetic characteristics of NM394 between single and multiple doses. No accumulation in plasma is observed after multiple doses of 264.2 mg per day for 7 d.

Determination of Naftopidil in Dog Plasma by High Performance Liquid Chromatography and Study on Its Pharmacokinetics

建立HPLC方法测定犬血浆中的萘哌地尔浓度，研究萘哌地尔胶囊在犬体内的药物动力学。单剂量给予5只健康犬萘哌地尔胶囊200mg,血浆样品碱化后，经乙醚提取以乙腈：磷酸盐缓冲液（0．05mol·L^-1的磷酸二氢钾溶液，以0．1mol·L^-1的NaOH调节pH至6．5）＝60：40（v/v)为流动相，由ODS C18分析柱分离测定，紫外230nm为检测波长，维拉帕米为内标。线性范围为10ng·L^-1;方法回收率为：100．23±3．00％；检测限：8ng·mL^-1;日间RSD≤5.56%,日内RSD≤3．30％。本法简便，回收率和灵敏度高，可用于萘哌地尔制剂的药动学研究。单剂量给予犬萘哌地尔胶囊200mg后血药浓度随时间变化规律符合一级吸收一室模型，T1／2Ke为3．19±1．27h,Tmax为1.15±0.59h,Cmax为697.48±94.22ng·mL^-1.

Determination of fleroxacin in human plasma by HPLC with fluorescence detection and the pharmacokinetic study

Aim To develop a sensitive and accurate HPLC method for the determination of fleroxacin in human plasma, and study its pharmacokinetics in healthy subjects. Methods The analytes were isolated fi＇om plasma by simple protein precipitation with methanol, separated on a Diamonsil C18 column by isocratic elution with the mobile phase consisted of 1% triethylamine at pH 4.8 （adjusted with phosphoric acid） and acetonitrile （80/20, V/V） at a flow rate of 1.0 mL.min^-1 and analyzed by fluorescence detector with an excitation at 290 nm and emission 458 nm. The pharmacokinetic study of fleroxacin was performed according to a double period crossover design. Results The weighted （1/x） calibration curve was linear over the plasma concentration range of 0.025 - 8.00 μg.mL^-1 The inter- and intra-day precisions （RSD/%） were no more than 5.16%, and the method accuracies and extraction recoveries at three concentrations ranged firom 99.1% to 100.9%, and 86.7% to 92.0%, respectively. Following oral administration at a dose of 400 mg fleroxacin, the main pharmacokinetic parameters for test and reference capsules were Cmax5.08 ± 0.78 and 5.38 ± 1.40 μg.mL^-1, tmax 1.72 ±0.79 and 1.82 ± 0.78 h, t1/2 11.68 ± 1.27 and 11.38 ± 1.51 h^-1, AUC0-∞ 78.44 ± 11.44 and 76.53 ± 13.24 μg.mL^-1.h, respectively. Conclusion The method is sensitive and accurate, and suitable for human pharmacokinetic study of fleroxacin.

Method for quantification of prazosin in dog plasma and its application to pharmacokinetic study

A simple and sensitive high performance liquid chromatography method using fluorescence detection （HPLC- FLD） and a one-step single solvent extraction for the determination of prazosin（PZS） in dog plasma is developed and validated. After extraction with ether, the chromatographic separation of PZS is carried out using a reverse phase C18 column （ 150 mm ×4.6 mm, 5 μm） with a mobile phase of 30% acetonitrile and 70% acetic acid-sodium acetate buffer solution （pH = 3.6） and quantified by fluorescence detection operated with an excitation wavelength of 258 nm and an emission wavelength of 387 nm. The flow rate of the mobile phase is 1.0 mL/min and the retention time of PZS and the internal standard is found to be 4. 4 and 5. 8 rain, respectively. The calibration curve is linear within a concentration range from 1.0 to 1 000.0 ng/mL （ P 〉 0. 998）. The limit of detection is 0.4 ng/mL. The inter-day coefficient of variation （COV） of the calibration standards is below 5.0% and the mean accuracy is in the range from 92. 7% to 104. 2%. Moreover, by analyzing quality control plasma samples for three days, the results show that the method is precise and accurate, for the intra- and inter- day COV within 10% and the accuracy from 95.9% to 112.7%. The developed and validated method is successfully applied to phannacokinetic study for the preclinical evaluation of a new peroral PZS-sulfobutyl ether beta-cyclodextrin （PZS-SBE-β-CD） inclusion complex tablets （test preparation）, which demonstrates that the test preparation released PZS is conducted in a slow and controlled way, and the relative bioavailability of the test preparation is found to be 105.0%.

Determination of Lansoprazole by Direct Injection of Plasma and High Performance Liquid Chromatography with Column Switching

High performance liquid chromatography with column switching has been developed for the determination of lansoprazole in plasma. The plasma samples were injected onto a pretreatment column packed with LiChromprep RP2 (25～40 mm) after simple dilution with distilled water. Distilled water was used to wash out protein and other polar components in plasma. After switching, the concentrated lansoprazole was eluted in the backflush mode onto a Shimpack CLC ODS column with methanol 0 2 mol·L 1 ammonium acetate (65:35) as mobile phase. Purge solutions were used for clean up and for regenerating the pretreatment column. The method showed good precision and recovery. The detection limit was 0 005 mg·L -1 plasma. The RSD’s (intra and interday) were less than 2 5% and 5 3% respectively. The method has been successfully used to determine pharmacokinetics of lansoprazole in Chinese volunteers.

Study of Linearization of Hill Dose-Effect Curve with Metabolic Velocity Instead of Drug Concentration

Objective To explore the velocity-effect relationship in order to the establish linearization of effect on an equation with regard to the consistency of the Hill dose-effect expression with the metabolic kinetics of receptors.Methods The linear velocity-effect expression was obtained by solving multivariant differential equation groups,which were established to compare the coincidences and basic relations between the Hill dose-effect and metabolic kinetic Michaelis-Menten equation for receptors.The validation test was conducted with acetylcholine,adrenaline,and their mixture as model drugs.Results The linear velocity-effect modelling was represented in vivo or in vitro,for single and multidrug systems.Pharmacodynamic parameters,especially suitable for multicomponent CMM formulas,could be determined and calculated for single or multicomponent formulas at high saturating or low linear concentration for receptors.The validation test showed that the pharmacodynamic parameters of acetylcholine were:k,2.675×10^-3s^-1;ka,5.786×10^-9s^-1;km,2.500×10^-7s^-1;α,4.619×10^9张s·mg^-1;E0,13张(P<0.01)and those of adrenaline were:k,1.415×10^-3s^-1;ka,5.846×10^-9s^-1;km,2.300×10^-7s^-1;α,-1.627×10^9张s·m g^-1;E0,9.2张(P<0.01).For the mixture of the two components,the values were:α,1.375×1010张s·m g^-1;-6.150×10^9张s m g^-1for acetylcholine and adrenaline,respectively,and E0was7.08张in both,with the other parameters unchanged(P<0.01).Conclusion The velocity-effect equation can linearize the Hill dose-effect relationship,which can be applied to study the pharmacodynamics and availability of CMM formulations in vivo and in vitro.

Determination of 2-Amino-6-Cyclopropylamino-9-(2,3-Dideoxy-β-D-glyceropent-2-enofuranosyl)purine in Rat Plasma,Urine and Liver Homogenates by High Performance Liquid Chromatography

Aim To develop a simple and specific high-performance liquid chromatographic(HPLC) method, suitable for the pharmacokinetic studies in vivo, to determine the concentrations of2-amino-6-cyclopropylamino-9-(2,3-dideoxy-β-D-glyceropent-2-enofuranosyl)purine (Cyclo-D4G, IMGprodrug) in rat plasma, urine and liver homogenates. Methods Chromatography was performed with C-18Hypersil ODS column and a mobile phase of 7% (v/v) acetonitrile in phosphate buffer, pH 7.40, withUV detection at 283 nm. Results The average extraction recovery of Cyclo-D4G in rat plasma and urinewas 100.1% over its linear range of 0.5 - 80 μg·mL^(-1). The accuracy of the assay was 99.4% .The intra-and inter-day RSDs were less than 9.0% . Conclusion The analytical method was found to beapplicable, reliable and suitable for pharmacokinetic studies.

Kinetics of water–isocyanate reaction in N,N-dimethylformamide

The uncatalyzed reaction of p-tolyl isocyanate(p-TI)with water in N,N-dimethylformamide(DMF)was investigated by high performance liquid chromatography(HPLC).The reactions were carried out at different temperatures from 293 K to 323 K,using various molar ratios of water to p-TI.DMF,as a special amide,was proved to be an efficient catalyst for water–isocyanate reaction.Under the reaction conditions in this study,substituted urea was the only final product observed.An appreciable amount of intermediate p-toluidine was detected.Concentrations of the isocyanate group as well as the amine and urea were determined as a function of time.New kinetic equations were deduced for each of the substance on the basis of a multistep mechanism,instead of a simple second order reaction as usual.Kinetic constants were calculated using the software MATLAB.Furthermore,the effects of temperature and concentrations of reactants on the reaction rate and amine content were discussed.The activation energy of each step was also determined.

Biodegradation of Acid Anthraquinone Dye in a Facultative-aerobic Process:Kinetics and Products

The transformation of an anthraquinone dye blue 324 in a facultative-aerobic(F-A) system was investigated.Kinetic parameter study showed that higher Vmax coupled with more recalcitrant chemical oxygen demand(COD) were found in the facultative biofilm reactor(FBR) than in the aerobic reactor(AR).Results of the product analyses indicated that most of dye molecular could be facultatively broken down into simple intermediates,which would be further degraded under subsequent aerobic condition.The main metabolites in each reactor were detected by infrared(FT-IR) and high performance liquid chromatography and mass spectrometry(HPLC-MS).Comparison of the toxicities among the dye and its metabolites was conducted,surprisingly,the colorless intermediates from FBR possessed less inhibitory than original dye and the median effective luminescence concentration(EC50) in 15 min for aerobic effluent could not be detected,showing that hardly toxic products existed in the aerobic process effluent.

中药燃烧焓、信息熵及生物熵的稳定性揭示其成分间的“虹势性”

目的:根据生物物理学原理探讨生物间代谢的"虹势性"普遍规律,为建立完整的中药物质质量控制方法奠定理论基础。方法:根据生物热力学定律建立中药材的信息熵等状态函数关系式,并以大黄药材醇浸出物为对象,建立指纹图谱,计算生物熵和表观平衡常数,探讨诸状态函数的稳定性,验证"虹热性"规律。结果:全国9地方的大黄醇浸出物的相似度为0.079～0.677;色谱峰面积的相对标准差(RSD)为21.11%～92.51%;而燃烧焓、信息熵、信息量、生物熵、表观平衡常数的RSD分别为2.259%,4.228%,20.190%,-2.254%,13.340%。结论:全国各地大黄醇浸出物的燃烧焓、信息熵、生物熵是稳定的。由中药成分的燃烧焓等参数可构成中药成分的宏观物质质量体系。

中药归经研究的现状及“穴药”法的提出

目的:分析中药归经研究的现状,提出并阐明"穴药"法的中药归经研究法。方法:对中药有效成分群(物)的归经及与经络脏腑功能(象)的互应关系的研究现状进行剖析,找出其不足与解决问题的对策。结果:特定的中药成分群对特定的经络脏腑有选择性的作用,这是中药归经的物质基础,体现"物"性;经络脏腑的功能状态以一定的证候表现出来,体现"象"性,一定功能的经络脏腑对特征中药成分群有反作用,可用于探知和表征经络脏腑,体现"候"性,三者实为"物-象-候"印迹关系。经络脏腑功能状态也可通过针刺特异腧穴改变,而这种改变可以通过中药定量药理学参数表征,据此可建立中药归经研究的"穴药"新方法,也可以探讨中药成分与病证网络靶点的关系,起到一箭双雕的作用。结论:中药与经络脏腑及其功能三者互为"物-象-候"印迹关系,可通过针刺特异性腧穴改变,"穴药"法既是中药归经研究最简捷的方法,也是探知经络脏象重要方法。