新型核苷类似物FNC在Beagle犬体内的血浆动力学及大鼠组织分布研究

发布时间：2018-04-04 13:20

本文选题：FNC　切入点：药代动力学　出处：《郑州大学》2014年硕士论文

【摘要】：FNC (2’-脱氧-2’-氟-4’-叠氮-核苷类似物)为郑州大学化学与分子工程学院研发的一种新型核苷类似物。药效学、毒理学研究表明，FNC毒性较低且具有很好的抗病毒、抗肿瘤作用。对胃癌、肝癌、肉瘤具有良好抗肿瘤活性[1,2]，具有很强的抗艾滋病毒（HIV）和乙肝病毒（HBV）活性[3-5]。FNC有望成为新型高效低毒的抗病毒、抗肿瘤核苷类药物。对FNC进行非临床药物代谢动力学研究有利于揭示FNC在动物体内的动态变化规律，可为药物制剂的特性和质量进行评价，对药物的研究开发起着重要作用，为指导临床合理用药提供参考信息。本实验室已经对FNC的药代动力学特征做了一些前期研究，包括FNC在啮齿类动物大鼠体内的血浆动力学、血浆蛋白结合率以及在大鼠体内的排泄研究，其结果表明FNC在大鼠体内生物利用度较高，灌胃给药后在大鼠体内过程符合一室模型，在有效剂量范围具有线性动力学特征，最后主要以原形及代谢物形式通过肾脏排泄[6]；血浆蛋白结合率较低，有利于药物快速分布发挥药效。为了全面评价FNC的药代动力学特征，需要进一步进行FNC在非啮齿类动物体内的药代动力学特征研究，因而本实验进行FNC在Beagle犬体内的血浆动力学以及在大鼠体内的组织分布研究。本研究主要分两部分，具体内容如下: 第一部分FNC在Beagle犬体内的血浆动力学研究第一节Beagle犬血浆样品中FNC HPLC-MS/MS定量分析方法的建立与验证采用甲醇蛋白沉淀法对血浆样品进行前处理，用HPLC-MS/MS方法对血浆样品中的FNC进行分离与检测，优化分析方法的色谱与质谱条件，建立犬血浆中FNC的测定方法并对本分析方法进行验证。结果显示：经过本实验建立的方法处理的样品FNC在0.5-400ng·mL-1线性关系良好（R20.99），定量限为0.5ng·mL-1，提取回收率在82.7%以上，日内、日间的RSD均小于7.49%。本方法灵敏度和准确度高，分析速度快，方法学验证结果符合体内药物分析要求，且稳定，能满足分析大量样品的需要。第二节FNC在Beagle犬体内的血浆动力学研究 Beagle犬单次经口灌胃给药FNC0.05、0.1、0.2mg·kg-1后，于不同时间点采血，血浆处理后用上述建立的HPLC-MS/MS法测定FNC的血药浓度，用DAS3.0软件采用非房室模型计算它的药代动力学参数。血浆中药物的消除半衰期t1/2z分别为(3.91±0.64) h、(3.61±1.04) h和(4.92±0.83) h；峰浓度Cmax分别为(45.77±5.08)、（80.47±8.88）和(200.57±34.11) ng·mL-1；AUC0-t分别为(227.53±23.98)、(375.85±74.84)和(1061.27±233.91) ng·h·mL-1， Cmax和AUC0-t均随剂量增加而增加(P0.05)，高、中、低剂量半衰期基本不变（p0.05），在0.05-0.2mg·kg-1范围内符合线性动力学特点，雌雄犬口服FNC的药代动力学不存在性别差异。第二部分FNC在大鼠体内的组织分布研究建立并验证FNC在大鼠组织中的HPLC-MS/MS分析方法，测定大鼠经口灌胃给药FNC制剂0.52mg·kg-1后组织中的FNC浓度，研究FNC在大鼠体内的组织分布情况。结果表明本实验建立的分析方法灵敏度和准确度高，方法学验证结果均符合体内药物分析要求，适合组织样品中FNC的分析检测。给药后FNC分布最多的组织是免疫器官胸腺和脾脏且消除较慢，胸腺中FNC含量在给药后12h达到最大，脾脏中FNC的浓度在给药后7h达到最大。在心、肝等组织分布较少且消除较快，在脂肪、脑和脊髓组织中含量最少。
[Abstract]:FNC ( 2 ' - deoxy - 2 ' - fluoro - 4 ' - azide - nucleoside analogue ) is a new nucleoside analogue developed by Zhengzhou University ' s Chemical and Molecular Engineering Institute . The pharmacodynamics and toxicology studies show that FNC has low toxicity and has good antiviral and anti - tumor effects . It has good anti - tumor activity for gastric cancer , liver cancer and sarcoma . It has a strong anti - HIV ( HIV ) and hepatitis B virus ( HBV ) activity . FNC is expected to be a new type of anti - virus and anti - tumor nucleoside medicament with low toxicity .

Non - clinical pharmacokinetic study of FNC is beneficial to reveal the dynamic regulation of FNC in animal body , which can provide reference information for the study and development of pharmaceutical preparations .
In order to fully evaluate the pharmacokinetic characteristics of FNC , it is necessary to study the pharmacokinetics of FNC in non - rodent . Therefore , the plasma kinetics of FNC in Beagle dogs and the distribution of tissue distribution in rats were studied .

This study is mainly divided into two parts , the specific contents are as follows :

Determination of FNC HPLC - MS / MS Quantitative Analysis Method in Beagle Dog Plasma Samples by First Part FNC Plasma Dynamics Study in Beagle Dogs

The plasma samples were subjected to pretreatment with methanol protein precipitation method . The separation and detection of FNC in plasma samples were carried out by HPLC - MS / MS . The results showed that the linearity of FNC in plasma samples was good ( 20.99 ) , the limit of quantification was 0.5ng 路 mL - 1 , and the RSD was less than 7.49 % .

The plasma concentrations of FNC were determined by HPLC - MS / MS method established by HPLC - MS / MS method . The elimination half - life t1 / 2z of drug in plasma was ( 3.91 卤 0.64 ) h , ( 3.61 卤 1.04 ) h and ( 4.92 卤 0.83 ) h , respectively .
Cmax were ( 45.77 卤 5.08 ) , ( 80.47 卤 8.88 ) and ( 200.57 卤 34.11 ) ng 路 mL - 1 , respectively .
The AUC0 - t was ( 227.53 卤 23.98 ) , ( 375.85 卤 74.84 ) and ( 126.27 卤 233.91 ) ng 路 h 路 mL - 1 , Cmax and AUC0 - t increased with increasing dose ( P0.05 ) .

Study on the Tissue Distribution of the Second Part of FNC in Rats

To establish and verify the HPLC - MS / MS analysis method of FNC in rat tissues , the concentration of FNC in the tissues of rats treated with FNC ( 0.52 mg 路 kg - 1 ) was determined .

【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R965

【参考文献】