【作者】 彭利霞；

【导师】 向楠；

【作者基本信息】 湖北中医药大学 ， 中医内科学， 2010， 博士

【摘要】 目的：通过菟丝子总黄酮药代动力学试验,探讨中药药代动力学研究思路与方法。方法：1.正交设计优选提取菟丝子总黄酮工艺。2.提取的菟丝子总黄酮进行健康志愿者口服,分别于0、0.5、1.0、2.0、3.0、4.0、8.0、12.0、24.0、48.0小时抽血2ml至肝素抗凝管中冷冻保存待测。将抽取的血浆室温融化,置lml离心管中,精密加入醋酸乙酯0.6ml,于涡流液体混合器上涡旋2min,离心5 min(3000 r／min-1),吸取上清液,37℃水浴氮气流挥干,残渣用40μl甲醇溶解。HPLC测定含量,‘进样20μl,记录峰面积进行定量分析。3.运用3p97药代动力学软件进行数据分析,分别计算不同房室数、权重系数,比较拟合优度值,得出相关药代参数。4.以菟丝子为例研究中药药代试验如成分复杂、缺乏化学对照品、有效成分不明确、试验操作质量难控制等问题,探讨如何在中医理论指导下如何进行中药药代动力学研究。结果：1.正交设计实验结果,提取工艺为A,B,C3D3,即乙醇浓度50%,加10倍量乙醇,提取90min,提取3次。乙醇浓度对金丝桃苷含量有显著影响(P<0.05),乙醇量、提取时间、提取次数对金丝桃苷的含量无显著意义(P>0.05),即因素A对试验结果有显著性影响,因素B、C、D对试验结果无显著性影响。影响金丝桃苷因素的顺序依次为A>D>C>B,即乙醇浓度>提取次数>提取时间>乙醇量,故在生产中、实验过程中要保证乙醇浓度,最佳制备工艺为A1B3C3D3,即乙醇浓度50%,加10倍量乙醇,提取90min,提取3次。2.药代试验,经药代动力学软件3p97分析试验数据,菟丝子总黄酮中的金丝桃苷在体内吸收代谢过程符合一室模型。达峰时间3.26h,分布半衰期1.55h,消除半衰期3.48h,说明菟丝子中的金丝桃苷在体内吸收与消除时间相当,提示菟丝子在临床应用中,给药间隔时间为约7小时,菟丝子在体内消除较快,长期服用不易产生蓄积现象,安全性高。3.试验中面临的问题及对策3.1中药成分复杂及有效成分不明确问题,中药药代动力学研究可从如下方面进行研究,其一：要研制大量化学对照品并建立指纹图谱,研制中药标准物质。由于中药的化学成分的复杂多样,这些研究还需要大量人力、物力,不是短期内能解决的。其二：示踪法。采用放射性同位素或稳定性同位素标记药物,并检测其在体内的吸收、分布、排泄动态过程,具有良好的灵敏度、较高的准确性、广泛的适用性,但放射性同位素法对环境有污染,稳定性同位素的标记物制作成本较高、难度较大。其三：PK／PD结合,以“效”对“证”,建立药效-时间关系,PK／PD结合模型能描述和预测一定剂量方案下药物的效应时间过程,科学地揭示药物剂量、相应时间与机体的效应关系,针对中药复方有学者提出“证治药动学”研究复方。从此思路进行研究的关键在于,确定中医诊疗标准,“证”和相应的“疗效评价”标准。综上述,中药药代动力学研究的关键问题是标准的问题,包括中医的标准和中药的标准。3.2中医诊疗技术过程,需要规范化、标准化、信息化。建立中医诊疗标准。方法学上,采用证候信息学、模糊数学、量表学、证候疗效评价等。中医证候信息学是将证候作为一种医学信息,运用现代信息技术加以采集、分析与处理,临床证候数据的采集是指将临床诊疗过程中的证候信息量(包括症状、体征、实验室检查等)收集后储存下来的过程,将证候客观化、规范化和数字化是中医证候学与信息学结合的基础与关键。中医药的标准化,首先确定标准体系框架。中医药标准体系的构建,能推动中医药标准化建设,对中医药标准化具有重大现实意义和作用,为中医药现代化和国际化创造了条件和提供标准支撑。3.3临床试验过程中质量可控制性的问题。中药临床试验还没有适合自己的完整详细指导原则,中药有其自身的特点照搬西药必然有不合理之处,解决中药临床试验面临的问题,首先制定体现中医药特点的中药临床试验指导原则及标准,据此建立适合中医临床操作的详细操作标准规程,便于操作人员理解执行,在管理上质量可控。提高中药临床研究质量的关键问题就是CRF的规范化、电子化,再构建适合中药的药物临床试验信息系统,对整个试验全过程动态监控,不仅可控制临床研究的质量结果,还可减轻研究者的工作并方便管理者管理。CRF的规范化、电子化的关键是中医诊断标准、疗效评价标准的建立4.中药药代研究思路上,脱离中医理论的指导的问题。应坚持中医理论为指导的原则,坚持中医药的整体观,遵循中药的组方规律,从整体、功能状态入手探求中医诊疗特色优势以及符合自身规律的临床疗效评价方法,把解决涉及中医药学术系统性和复杂性等关键问题作为中医药研究的主要任务和目标；发展适用于复杂生命系统研究和定量化分析的新技术和新方法,系统地提升中医药学理论和中医药研究能力。结论：1.菟丝子提取工艺为乙醇浓度50%,加6倍量乙醇,提取90min,提取3次。菟丝子提取物金丝桃苷口服给药后,在体内体内过程符合一室模型。达峰时间3.26h,分布半衰期1.55h,消除半衰期3.48h,提示菟丝子在体内消除较快,长期服用不易产生蓄积现象。2.中药药代动力学作为一门紧密联系中医与中药的桥梁学科,目前面临的关键问题是标准。即不仅需要中医的标准,还需要中药的标准。在此基础上,做中药作用机理研究,指导临床合理用药,为中医药临床疗效有效性、安全性提供客观依据。3.中药药代动力,学研究思路,坚持中医理论为指导的原则。更多还原

【Abstract】 Objective:By Cuscuta chinensis Lam.total flavonoids from pharmacokinetic trial,I discussion pharmacokinetic study of traditional Chinese medicine ideas and methods.Method:1.Orthogonal design experimental Optimal extraction Cuscuta chinensis Lam.total flavonoids craft。2.The healthy volunteer takes Cuscuta chinensis Lam.total flavonoids.at 0、0.5、1.0、2.0、3.0、4.0、8.0、12.0、24.0、48.Ohour, Draws blood to freeze the preservation 2ml to the heparin antifreeze tube in testing.Will extract blood plasma room temperature melting;Sets at 1ml in the centrifuge tube; Joins ethyl acetate 0.6 ml precisely;On turbulent flow liquid mixer vortex 2min;Centrifugal 5 min(3000 r/min-1);Absorption supernate;37℃the water bath nitrogen air current wields does;The residual dissolves with 40μl the methyl alcohol。HPLC determination content,Specimen handling 10μl; The record peak area carries on the quantitative analysis。3.The use of 3p97 software analysis pharmacokinetic data;Calculate the number of different atrioventricular,Weight coefficient,Goodness of fit value;obtained relevant pharmacokinetic parameters.4.To the problems encountered during the experiment:Dodder lack of standard components and component complex, active ingredient is not clear, difficult to control the quality of test operationOther issues.Discussion solution countermeasure, how does conduct the traditional Chinese medicine medicine generation of dynamics research under the Chinese medicine theory instruction.Results:1.Orthogonal design experimental result:The extraction process is A1B1C3D3,Ethyl alcohol density 50% adds 10 time of quantity ethyl alcohol to withdraw 90min, withdraws 3 times. The ethyl alcohol density has the remarkable influence to the Hyperoside content(P<0.05).Ethyl alcohol quantity, extraction time,extraction number of times to Hyperoside content not remarkable significance(P>0.05).It is that factor A has the significance influence to the test result, Factor B, C, D to test result non-significance influence.The influence Hyperoside glucoside factor’s order is A>D>C>B in turn.The ethyl alcohol density>withdraws the number of times>to withdraw the time>the ethyl alcohol quantity. Therefore in the production and the experiment process must guarantee the ethyl alcohol density. Best preparation craft is A1B3C3D3,the ethyl alcohol density 50%, add 10 time of quantity ethyl alcohol,withdraws 90min,withdraws 3 times.2.Pharmacokinetics Test:the pharmacokinetic analysis of data software 3p97, the hyperin In total flavonoids sbsorption metabolism in the body consistent with one compartment model.T(peak)3.26h,t1/2(ka)1.55h, t1/2(ke)3.48h,Shows hyperin Cuscuta in vivo absorption and elimination in the time gap.3.In the experiment faces question and countermeasure:3.1 the chemical composition of traditional Chinese medicine complex and diverse。first:to develop a large number of standards、Traditional Chinese medicine standard substance and establish fingerprint.Since the chemical composition of traditional Chinese medicine complex and diverse,.Development of standard also require substantial human,material.in the short term can not be resolved.second, Tracer technique.Using radioisotope or stable isotope labeled drug, and determine the in absorption, distribution,excretion dynamic process,with good sensitivity, high accuracy, wide applicability, but the radioactive pollution of the environment law, Stable isotope’s tracer manufacture cost is high,the difficulty to be big.Third, PK/PD union.to "effect" on the "certificate",the establishment of efficacy-time, Some scholars put forward the "Pharmacokinetics and Treatment" of compound.In this respect, need medicine "certificate" and "efficacy evaluation" standards.Combination of the two to see, Need to develop a series of standards, The standard of Chinese medicine, Chinese medicine quality standards, including standards for Chinese medicine products, fingerprinting, diagnostic criteria for traditional Chinese medicine, TCM clinical efficacy evaluation criteria.Pharmacokinetics of traditional Chinese medicine the key issues is a standard problem,traditional Chinese medicine, including standards and criteria.3.2 Chinese medicine treatment technology process.,need standardization,information.Establish standards for Chinese medicine clinics. Methodology, using the syndrome information science, mathematics,science scale, syndromes evaluation.TCM Syndrome informatics is as a medical information,The use of modern information technology to collect, analyze and deal with.Clinical data acquisition is collected and stored procedure.in the diagnosis and treatment the amount of information(including symptoms, signs, laboratory tests, etc.).The syndrome of objective, standardized and digital is the foundation and the key of integration the syndrome and information science.The standardization of Chinese medicine,first determine the standard system framework. Standard System of Traditional Chinese Medicine, promote the standardization of Chinese medicine, has great practical significance and role for standardization in Chinese medicine,create the conditions and standards of support provided for the modernization and internationalization of Chinese medicine.3.3 The quality of clinical trials can be controlled The traditional Chinese medicine clinical trial has not suited own complete detailed guiding principle. the traditional Chinese medicine has its own characteristic.The traditionalChinese medicine clinical trial copy western medicine to have the unreasonable place inevitably.the question which the solution traditional Chinese medicine clinical trial faces, first formulates manifests the Chinese medicine characteristic the traditional Chinese medicine clinical trial guiding principle and the standard.On this basis, we establishes suits the Chinese medicine clinical operation the detailed operation standard regulations.It is advantageous for the operators to understand the execution.the quality is controllable in the management.The key question is the CRF standardization,computerization improves the traditional Chinese medicine clinical research quality.To construct a suitable the traditional Chinese medicine the medicine clinical trial information system,to monitor entire experimental process,not only we may control the clinical research the quality result, but may also reduce researcher’s work and the convenient superintendent manages.The CRF standardization,the computerization key is the Chinese medicine diagnosis standard,the curative effect evaluation criteria establishment.4.In the traditional Chinese medicine medicine generation of research mentality,it is separated from the Chinese medicine theory the instruction question.It Should adhere to the guiding principles theory of the Chinese medicine, the total perspective of the Chinese medicine, follows the group side rule of the traditional Chinese medicine.From the whole function condition to seek the superiority of the Chinese medicine diagnosing and treating characteristic as well as to conform to own rule clinical curative effect assessment method.It is the main objective to resolve the qusetion of the Chinese medicine academic systematic and complex.The new technology and the new method are developed suitable for the complex life system research and the quantitative analysis.To promotes the Chinese medicine pharmacy theory and Chinese medicine research ability systematically. Conclusion:1.Cuscuta chinensis Lam extraction process for ethyl alcohol density 50%, Adds 6 time of quantity ethyl alcohol, Withdraws 90min,Withdraws 3 times.After the Cuscuta chinensis Lam extraction oral administration gives the medicine, Hyperoside conforms to the one room of model in vivo in vivo process.Reaches peak time 3.26h, Distributed half-life 1.55h,Eliminates half-life 3.48h.The cuscuta eliminates quickly,takes for a long time is not easy to produce stores up the phenomenon,the security is high.2.The pharmacokinetics of traditional Chinese medicine as a close contact with the bridge disciplines of Chinese medicine, the key issues currently facing is the standard.That demand not only the standard of Chinese medicine, but also the standard of Chinese medicine.On this basis, it provides the objective basis for the Chinese medicine clinical curative effect validity, the security.the traditional Chinese medicine action mechanism research,the instruction clinical reasonable medication.3.Pharmacokinetics of traditional Chinese medicine ideas, it adhere to the principles of Traditional Chinese Medicine Theory.更多还原