Antiepileptic drug therapy is a main method for controlling epilepsy,but the responses of patients to the current treatments are not consistent due to inter-individual differences in drug disposition.In the present study,we investigated whether genetic and epigenetic variants affected the pharmacokinetic phenotypes of the antiepileptic drug 3,4-dichlorophenyl-propenoyl-sec-butylamine(3,4-DCPB)in phase I dose-escalation clinical trial in healthy subjects.The plasma concentrations of 3,4-DCPB and its major metabolite Ml were determinated by the liquid chromatography tandem mass spectrometry(LC-MS/MS)method.Single nucleotide polymorphisms(SNPs)of xenobiotic metabolisms including cytochrome P450 2D6(CYP2D6),CYP2C9,CYP1A2,CYP2C19,CYP3A5,transporter ABCB1(C1236T),nuclear receptors AhR,CAR and PXR were analyzed by genotyping and DNA methylation levels for these genes.Compared to the wild-type CYP2D6*1/*1 homozygote(extensive metabolizers,EMs),the variant allelic CYP2D6*10 carriers(intermediate metabolizers,IMs)showed that the area under the curve(AUC0-t)ratios of metabolite Ml/3,4-DCPB parent drug were lower,and the plasma half-life(t1/2)ratios were prolonger,while the DNA methylation levels were higher.These data suggested that epimutation induced by lose(CYP2D6*10,C＞T)of cytosine,might explain the associations among genotype,epigenotype and individual differences in the pharmacokinetic phenotype of 3,4-DCPB,and provide new insight in personalized treatment of epilepsy.

Objective To test the acute toxicity and determine the safe dose range of Moringa oleifera leaves and seeds.Methods Acute toxicity tests were performed based on the documented experimental designs in Research Methods of Pharmacology of Chinese Traditional Medicine.Results The maximum tolerated dose of the superfine powder and alcoholic-aqueous extract of Moringa oleifera leaves were 0.4 g/20 g and 0.8 g/20 g for mice.The safe dose for human is under or equal to 10 g/50 kg and 20 g/50 kg.Conclusion Moringa oleifera leaves were safe for medical and food use.The further research and development is warranted for this plant.

We aimed to evaluate the bioequivalence of clopidogrel in healthy Chinese volunteers after administration of a single oral dose. We administered a single oral dose of 75 mg clopidogrel (test and reference) to 32 healthy Chinese volunteers according to an open, randomized, crossover design. The concentration of clopidogrel acid (carboxylic metabolite of clopidogrel) in the plasma was determined using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Bioequivalence of the test and reference preparations were calculated using analysis of variance and one-sided t-test by using the DAS 2.0 software. The pharmacokinetic parameters of the test and reference preparations were as follows: peak plasma concentration (Cmax), 1351.101 ± 654.955 ng/mL and 1184.652 ± 607.713 ng/mL; area under the curve, 2642.017 ± 1093.848 ng·h/mL and 2780.666 ± 1283.100 ng·h/mL; and time to reach Cmax (Tmax), 0.789 ± 0.318 h and 0.953 ± 0.633 h, respectively. The relative bioavailability of the formulation was 101.7 ± 35.3%, which indicated that the test preparation was bioequivalent to the reference drug.

目的 研究厄多司坦胶囊在健康人体的药代动力学,评价其生物等效性.方法 20名男性志愿者随机分为2组,按照两制剂两周期的随机交叉试验设计,分别单剂量口服厄多司坦试验或参比制剂各1粒(0.3 g),采用LC-MS/MS法测定血浆中厄多司坦及其代谢产物M1的浓度,用DAS2.0药动学软件分别计算厄多司坦及其代谢产物M1药动学参数并进行生物等效性统计分析.结果 试验及参比片的厄多司坦的主要药动学参数Cmax分别为(1570.7±488.3)和(1447.8 ±449.9) ng ·ml-1;Tmax分别为(1.1±0.5)和(1.2±0.5)h;t1/2分别为(1.3±0.3)和(1.3±0.4)h;AUC(0-10h)分别为(3444.7±1242.4)和(3229.9±1225.6)ng·h ·ml-1;AUC(0-∞)分别为(3485.0±1233.7)和(3279.6±1233.7) ng·h·ml-1;双单侧t检验结果表明,厄多司坦受试制剂Cmax的90％置信区间落在参比制剂的94.1％～124.4％范围内,AUC(0-10h)的90％置信区间落在参比制剂的90.7％ ～ 123.9％范围内;受试制剂对参比制剂厄多司坦的相对生物利用度F为(116.6±36.6)％.试验及参比片的M1的主要药动学参数Cmax分别为(396.5±177.6)和(385.0±192.7) ng·ml-1;Tmax分别为(1.7±0.8)和(1.9±0.7)h;t 1/2分别为(2.6±0.9)和(2.6±1.1)h;AUC(0-10h)分别为(1259.2±609.8)和(1331.3±694.0)ng·h·ml-1;AUC(0-∞)分别为(1372.4 ±607.3)和(1451.7 ±719.0) ng·h·ml-1;双单侧t检验结果表明,M1受试制剂Cmax的90％置信区间落在参比制剂的89.2％ ～ 121.1％范围内,AUC(0-10h)的90％置信区间落在参比制剂的80.9％ ～111.6％范围内;受试制剂对参比制剂M1的相对生物利用度F为(103.2±39.6)％.结论 对厄多司坦及其代谢产物M1的主要药动学参数进行评价,按照生物等效性判定标准,认为2种制剂生物等效.

目的 评价人ALDH2基因多态性检测试剂盒(荧光PCR法)用于人ALDH2基因c.1510位点(G/A)核苷酸多态性检测的准确性和有效性.方法 将入选人基因组DNA标本371例采用金标准和临床待考评试剂盒进行同步盲法检测,试验结束后揭盲,将试验试剂检测结果与金标准检测结果进行统计比较分析.结果 人ALDH2基因多态性检测试剂盒(荧光PCR法)与DNA直接测序法(Sanger测序法)金标准检测结果相比,两者基因型总符合率为99.19％,临床诊断评价总符合率99.46％,有较高的一致性.结论 人ALDH2基因多态性检测试剂盒(荧光PCR法),无放射性危害,具有高灵敏性、高特异性、试剂周期长等特点,可以在临床检验中推广使用.

目的:比较三种生姜的提取方法,优化生姜药材的提取工艺及工艺参数.方法:建立反相高效液相色谱法测定生姜指标成分6-姜辣素的分析方法,比较水蒸气蒸馏法、超临界CO2萃取法和乙醇渗漉法提取生姜的工艺.结果:该6-姜辣素的含量测定方法稳定可靠;超临界CO2萃取法和乙醇渗漉法提取率较高,超临界CO2萃取物中6-姜辣素的含量显著高于水蒸气蒸馏法和乙醇渗漉法.结论:三种方法中以超临界CO2萃取法为提取生姜的最佳方法.

Objective: To investigate and eliminate the factors affecting the stability of dexamethasone in Shengfu frost,and optimize its formulation. Methods: The dispersible uniformity test and the high temperature destruction test were carried out for quantitative analysis of the factors affecting the stability of dexamethasone, on the basis of which we optimized the formulation and process. Results: The study showed the content of dexamethasone would decrease apparently, when the pH level of the matrix was higher than 7.3. The solubility of dexamethasone was low in highly polar solvents, which was the main factor of heterogeneous dispersion. Dissolving dexamethasone in appropriate amount of laurocapram before the preparation will make the dispersion uniform. The content of dexamethasone could be stable when the pH level of the matrix was between 5.5 and 6.5. Conclusion: The accelerated destruction test and the solubility test were adopted for comprehensively analyzing the factors which affected the formulation stability. The method is reasonable and suitable for the quick investigation and optimization of formulations.

Objective To study the phosphorylation mode of extracellular regulated kinase( ERK)induced by acute and chronic morphine treatment on Chinese hamster ovary( CHO)cells expressed withμopioid receptors. Methods The time course of ERK phosphorylation 1 h and 36 h after morphine exposure as well as naloxone-precipitated withdrawal was detected by immunobloting. Results A transient enhancement of ERK phosphorylation was induced by 1 μmol · L-1 morphine with the peak effect at 5 min(P〈0. 01),and the effect was dose-dependent. No difference in ERK phosphorylation was found after 36h of treatment with 10 μmol · L-1 morphine compared with the control. However,5 or 10 min-naloxone precipitation induced remarkable decrease in ERK phosphorylation compared with the control(P〈0. 01). Conclusion Different changes of ERK phosphorylation were found under acute and chronic morphine treatment and naloxone precipitation,indicating a compensation of ERK related pathway induced byμ opioid receptors.

有机阴离子转运多肽1B1(OATP1B1)是肝脏中重要的药物转运通路之一,其遗传多态性可改变药动学参数,影响多种药物在体内的分布,继而影响药效及药物不良反应。该文对OATP1B1结构与功能,OATP1B1编码基因的结构及多态性对功能的影响,SLCO1B1多态性对他汀类药物的影响,SLCO1B1多态性对口服降糖药的影响,SLCO1B1多态性对抗肿瘤药的影响等进行综述。

Objective To investigate the effect of Tongxuan Lifei pill on CYP2D6and to provide foundation for clinical drug using.Methods Thirty volunteers were divided into two groups according to CYP2D6gene type with dextromethorphan used as probe drug of CYP2D6.The effect of Tongxuan Lifei pill on the activity of CYP2D6was investigated.Genetic classification was fulfilled by pyrosequencing method,and the blood concentration of probe drug and its metabolic product was determined by HPLC-MS/MS.The volunteers took Tongxuan Lifei pill for two weeks and AUC0~12 was used as the standard of the activity of CYP2D6.Results The activity value was 6.1±1.5before administration of drug and 6.5±1.7after drug using.The activity of CYP2D6was decreased after drug using,t-test was used for comparison,there existed no remarkable difference between the data.Conclusion Tongxuan Lifei pill has no obvious affection on the activity of CYP2D6.

Objective To investigate the effect of CYP2C9 polymorphism on the pharmacokinetics of meloxicam in Chinese healthy subjects.Methods The CYP2C9 genotypes of twenty-four Chinese healthy male subjects were selected by the pyrosequencing method.The subjects were genotyped with CYP2C9*3 wild-type and mutation.HPLC-MS-MS method was used for the determination of meloxicam in plasma.The important pharmacokinetic parameters were caculated by DAS 2.0 software.The pharmacokinetics of meloxicam based on identification of genotypes(CYP2C9*3 wild-type and mutation) was compared.Results Two kinds of CYP2C9*3 phenotype existed in the subjects.CYP2C9* 3(A/A) was found in twenty-two subjects while CYP2C9*3(A/C) in two subjects.For meloxicam,the A/A of CYP2C9*3 alleles was more active than the A/C.Conclusion CYP2C9 genetic polymorphism has significant influence on the pharmacokinetics of meloxicam.A/A is more active than A/C.Pharmacogenomic studies will help rational and individualized medication.

Objective To study the quality control of Lianqiaopikang Ointment.Methods TLC was used to identify Rheum officinale Baill,Radix et rhizome Clematidis and dexamethasone.HPLC system was used to determine the content of emodin,chrysophanol and dexamethasone in Lianqiaopikang Ointment.Results The spots of emodin,chrysophanol and dexamethasone of the test sample were consistent with the control.The linear range of emodin and chrysophanol was both 1-80 μg/ml,r=0.9999.The average recovery was 100.83% and 101.70%,respectively,and the relative deviation(RSD) was 3.03% and 3.51%,respectively.The linear range of dexamethasone was 5-80 μg/ml,and the average recovery rate was 100.36%,RSD was 1.54%.Conclusion The methods of TLC and HPLC used in this study are simple and accurate for quality control of Lianqiaopikang Ointment.

OBJECTIVE To study the pharmacokinetics and pharmacodynamics of amlodipine in healthy subjects and explore the relationship between the blood concentration of amlodipine and the antihypertensive effect. METHODS 24 male volunteers were administered 10 mg amlodipine by single dose.The blood samples of different time points in these subjects were sampled and the drug concentrations of amlodipine were analyzed by LC-MS-MS,and the primary variables measured in this study were systolic and diastolic blood pressure(SBP and SDP) and pulse rate in a sitting position. RESULTS Amlodipine was a two-compartment model with slow elimination(t1/2,36 h) and a large apparent volume of distribution.The Cmax was(6.1±1.5)ng·mL-1.SBP and SDP were depressed after administration,and it was related to the change of drug concentration.The efficacy hysteresis rings were found from the curves of efficacy-concentration. CONCLUSION Amlodipine can depress the SBP and DBP with efficacy hysteresis.The most efficacy Emax(SBP:6.3±3.2 mmHg,DBP:10.1±2.9 mmHg) was appeared after administration 6-10 h.The efficacy hysteresis was better form on the antihypertensive effect in clinical therapy.

目的建立盐酸丁螺环酮贴剂的质量控制方法。方法以紫外法进行定性鉴别,以初黏力、持黏力和剥离强度为指标控制黏度,采用高效液相色谱法(HPLC)对其含量均匀度、释放度进行测定。结果3批测试样品显示,紫外光谱鉴别在237和300nm波长处有最大吸收峰,初黏力为9号钢球全部在测试段黏住,持黏力为(647.00±7.81)s;剥离强度为(0.57±0.01)kN.m-1,含量均匀度(7.00±3.17)%,6h平均累积释放量(91.36±3.06)%。结论该检测方法科学、合理,操作灵敏、准确,可用于该药质量控制。

OBJECTIVE To study the percutaneous permeability of buspirone hydrochloride hydrophilic and hydrophobic patch in vitro.METHODS The influence of azone and urea in buspirone hydrochloride patch on its permeability through mice skin was studied in vitro.RESULTS Azone could enhance buspirone hydrochloride permeability effectively in mice skin.The percutaneous permetability rate of buspirone hydrochloride with 2% or 5% azone was steady after 4 hours.CONCLUSION Buspirone bydrochloride hydrophobic patch had a good percutanous permeability.The patch with 2% Azone was proved as a safe,economical and effective agent.

Objective To investigate the pharmacokinetics of ambroxol and clenbuterol tablets in Chinese healthy volunteers after a single-or multiple-dosage oral administration.Methods A total of 9 healthy adult subjects were given ambroxol and clenbuterol tablets in a single dosage or multiple dosage respectively.Liquid chromatography/tandem mass spectrometry was used for the determination of ambroxol and clenbuterol in plasma.The important pharmacokinetic parameters were caculated by DAS 2.0 software(compartment model).Results Both the single and multiple-dosage group of ambroxol and clenbuterol were suited to the two-compartment model.No difference in pharmacokinetics of ambroxol in groups of volunteers was observed,suggesting that the multiple dosage did not influence the velocity of drug metabolism.Moreover,parameters of clenbuterol was significantly different between the two groups(P<0.05),showing there was accumulation in the body.Nine subjects completed the single-or multiple-dosages oral administration test,with no adverse drug reactions during the test.Conclusion There is no obvious accumulation of ambroxol after repeated dosage while obvious accumulation of clenbuterol was noted in the multiple-dosage administration.The established method is sensitive,accurate,reliable and specific,and can meet the requirement of clinical pharmacokinetic trial.

OBJECTIVE To study the absorption of the buspirone hydrochloride transdermal paste in vivo and the method of assay in plasma. METHODS HPLC was used to determine the content of buspirone hydrochloride in rabbit plasma.RESULTS The detection vavelength of HPLC was 237 nm, acetonitrile-methanol-water-0.1 mol·L~(-1) sodium acetate buffer (pH 5.0) (38:10:51:1) was used as the mobile phase and there was a good linear relationship of hydrochloric acid in plasma in the range of 1-100μg·L~(-1); buspirone hydrochloride transdermal delivery patches have good skin permeability. After 24 hours of single administration of 12 mg·kg~(-1), the concentration of the drug in plasma was at steady fluctuations within a certain range. CONCLUSION The method is sensitive,accurate and good reproducible for the determination of the plasma concentra-tion of buspirone hydrochloride. This will provide scientific references for the rational transderrnal drug delivery formulations and the design evaluation in vivo.

Objective To study the percutaneous permeability of two kinds of nanoargentum hydrogels with different content of cellulose and phosphate as filling stuff in vitro. Methods Isolated rat skin was taken as permeable barrier,the skin permeability of nanoargentum in each hydrogels were observed by HS-vitro apparatus. Results Permeability accelerant increased the argentum content in skin effectively and cellulose lasted the action of nanoargentum. Conclusion 0.1% cellulose nanoargentum hydrogels have a good permeability and stable release property,which is benefit for local therapeutic application.

Objective To study the perceutanous absorption of homoharringtonine ointment in vitro. Methods Rats skin was applied as absorption barrers to study influence of azone at different concentrations on homoharringtonine permeation. Results Azone enhanced skin permeation of homoharringtonine significantly.Compared with the blank ointment,the steady percutanous absorption rate of homoharringtonine ointment containing with 2% and 5% azone increased by 56.88% and 89.51%,respectively. Conclusion Homoharringtonine cream have a good percutanous permeability,and 5% azone is the best permeating accelerator.

Aim To investigate the pharmacokinetics of prulifloxacin tablets in Chinese healthy volunteers after a single oral administration of prulifloxacin tablets.Methods A total of 12 healthy adult subjects were randomly grouped by 3×3 Latin square and assigned to receive a single oral dose of 100,200 and 400mg of prulifloxacin tablets.LC/MS/MS were used for the determination of NM394,the metabolite of prulifloxacin,in plasma after a single oral dose of prulifloxacin tablets.Important pharmacokinetic parameters were caculated by DAS 2.0 software(compartment model).Results No prulifloxacin but its metabolite-NM394 was identified in the blood sample of subjects.All the three dosage groups(100,200,400mg) conformed to the two-compartment model.The pharmacokinetics fitted first-order kinetic process.No difference in pharmacokinetics of NM394 in male and female volunteers was observed.12 subjects completed the single oral administration test,with no adverse drug reactions during the test.Conclusion The Cmax 、AUC of NM394 are highly correlated with given prulifloxacin doses.The established method is sensitive,accurate,reliable and specific,and it can meet the requirement of clinical pharmacokinetic trials.