国家一类肝癌治疗药BZG的临床前药代动力学和组织分布研究

发布时间：2018-09-13 06:34

【摘要】：背景:肝细胞肝癌是最为常见的肝脏原发性肿瘤,具有发现迟、恶性程度大、死亡率高等特点。索拉菲尼是目前肝癌治疗的唯一一线分子靶向药物,研究表明其能够增加晚期肝癌患者的总体生存率。但索拉菲尼价格昂贵,每位使用该药物的患者每个月的药品支出需要数万元人民币,给患者家庭造成较大的经济负担,此外索拉非尼还存在个别严重副作用等缺点,因此对其进行合理的结构改造或研制具有相似活性位点的药物已成为药物化学领域研究的热点。BZG是一种化学结构类似于索拉菲尼的多靶点酶抑制剂,我们既往的研究证实,它可以在体外抑制肿瘤细胞的增殖,显著抑制肝细胞肝癌异种移植模型中的肿瘤生长。目的:本课题在我们前期研究工作的基础上,拟应用临床前体内模型,阐明BZG吸收、分布性质,明确其临床前吸收代谢动力学性质,为其临床研究做好前期研究工作。本研究的最终目的是使BZG成为具有自主产权的一类新药,为我国广大肝癌患者提供新的药物选择,从而减轻患者治疗所需的经济负担,进而获得良好的经济效益和社会效益。方法:本文采用UPLC-MS/MS在合适的色谱和质谱条件下,利用伊马替尼作为内标,建立BZG在生物样品中的测定方法。根据FDA生物分析方法验证指南验证其专属性、线性、准确度、精密度、回收率、基质效应和稳定性。大鼠按照20mg/kg灌胃给药后,在0至48小时内的不同时间点取血液样本,按照所建立的生物样品中BZGUPLC-MS/MS测定法进行测定,取得给药后不同时间点的大鼠血浆中BZG的浓度,用DAS软件分析获得药代动力学参数。大鼠按照20mg/kg灌胃给药后,1,6和12小时收集以下组织:小肠、肝脏、肺、肾脏、胰腺、肌肉、脂肪和大脑,按照所建立的生物样品中BZGUPLC-MS/MS测定法进行测定,取得给药后不同时间点的大鼠不同组织中BZG的浓度,并据此研究药物的组织分布。结果:本文成功建立BZG在生物样品中的UPLC-MS/MS测定方法,其相关验证结果如下:在研究浓度范围(0.5ng/ml-2500ng/ml)内,BZG具有良好的线性(r2≥0.994);BZG色谱图中样品峰无杂质峰干扰,专属性良好。低、中、高浓度的质控样品的日间、日内准确度为-11.65%至11.76%,精密度为0.34%至14.64%;回收率分别为 80.42%至 89.46%,RSD10.87%;基质效应 84.62%至 92.41%,RSD≤10.31%;常温保存8h、-20℃保存40天、反复冻融3次以及处理后24h内的BZG稳定性在±15%之间。大鼠按照20mg/kg灌胃给药后,用DAS软件分析血药浓度的主要结果如下:大鼠体内的药代动力学行为符合一室模型;BZG在6.00±0.35小时达到587.08±84.08ng/ml的平均最大浓度;消除半衰期(t1/2)为2.86±2.49小时。大鼠按照20mg/kg灌胃给药后,其重要脏器的组织分布主要结果如下:BZG能够快速而广泛地分布到大鼠各组织中;所有组织均在6小时达到最高浓度;小肠内BZG浓度最高;然后依次是肾脏、肝脏、肺、胰腺、肌肉、脂肪和大脑。结论:1.本文建立并验证一种测定大鼠生物样品中BZG浓度的UPLC-MS/MS方法。2.按照20mg/kg灌胃后,大鼠体内的BZG在6.00±0.35小时达到587.08±84.08ng/ml的平均最大浓度;消除半衰期(t1/2)为2.86±2.49小时,符合一室模型。3.按照20mg/kg灌胃后,BZG能够快速而广泛地分布到大鼠各组织中,组织浓度由高至低依次为小肠、肾脏、肝脏、肺、胰腺、肌肉、脂肪和大脑。
[Abstract]:BACKGROUND: Hepatocellular carcinoma (HCC) is the most common primary liver tumor with the characteristics of late detection, high malignancy and high mortality. Sorafenib is the only first-line molecular targeted drug for the treatment of HCC. Studies have shown that it can increase the overall survival rate of patients with advanced HCC. The monthly drug expenditure of the patients needs tens of thousands of RMB, which brings great economic burden to the patients'families. In addition, there are some serious side effects of sorafenib. Therefore, it has become a hotspot in the field of pharmaceutical chemistry to modify the structure of sorafenib reasonably or to develop drugs with similar active sites. Our previous studies have confirmed that it can inhibit the proliferation of tumor cells in vitro and significantly inhibit the growth of hepatocellular carcinoma xenografts. OBJECTIVE: Based on our previous work, we intend to use a preclinical in vivo model to elucidate the absorption and distribution of BZG. The ultimate goal of this study is to make BZG a new class of drugs with independent property rights, to provide new drug options for the vast number of patients with liver cancer in China, so as to reduce the economic burden of patients with treatment, and to achieve good economic results. METHODS: UPLC-MS/MS was used to establish a method for the determination of BZG in biological samples by using imatinib as internal standard under suitable chromatographic and mass spectrometric conditions. After intragastric administration, blood samples were taken at different time points from 0 to 48 hours and determined by BZGUPLC-MS/MS. The concentration of BZG in rat plasma at different time points after administration was obtained. The pharmacokinetic parameters were analyzed by DAS software. After intragastric administration of 20mg/kg, the blood samples were collected at 1, 6 and 12 hours. The following tissues: small intestine, liver, lung, kidney, pancreas, muscle, fat and brain were determined by BZGUPLC-MS/MS. The concentration of BZG in different tissues of rats at different time points after administration was obtained and the tissue distribution of BZG in biological samples was studied. The relative validation results were as follows: within the range of 0.5ng/ml-2500ng/ml, BZG had good linearity (r2 < 0.994); sample peaks in BZG chromatogram had no interference from impurity peaks, good specificity. Low, medium and high concentrations of quality control samples during the day, the intra-day accuracy was - 11.65% to 11.76%, precision was 0.34% to 14.64%; recovery rate was 0.34%. The stability of BZG in rats was between (+) 15% after repeated freezing and thawing for 3 times and 24 hours after treatment. In order to conform to the one-compartment model, BZG reached 587.08 (+ 84.08ng/ml) at 6.00 (+ 0.35) hours and the elimination half-life (t1/2) was 2.86 (+ 2.49) hours. CONCLUSION: 1. A UPLC-MS/MS method was established and validated for the determination of BZG concentration in biological samples of rats. 2. After intragastric administration of 20mg/kg, the average BZG concentration in rats reached 587.08+84.08ng/ml in 6.00 (+ 0.35) hours. The elimination half-life (t1/2) of BZG was 2.86 (+ 2.49) hours, which accorded with one-compartment model.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.7

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