【摘要】：背景:肝細(xì)胞肝癌是最為常見(jiàn)的肝臟原發(fā)性腫瘤,具有發(fā)現(xiàn)遲、惡性程度大、死亡率高等特點(diǎn)。索拉菲尼是目前肝癌治療的唯一一線分子靶向藥物,研究表明其能夠增加晚期肝癌患者的總體生存率。但索拉菲尼價(jià)格昂貴,每位使用該藥物的患者每個(gè)月的藥品支出需要數(shù)萬(wàn)元人民幣,給患者家庭造成較大的經(jīng)濟(jì)負(fù)擔(dān),此外索拉非尼還存在個(gè)別嚴(yán)重副作用等缺點(diǎn),因此對(duì)其進(jìn)行合理的結(jié)構(gòu)改造或研制具有相似活性位點(diǎn)的藥物已成為藥物化學(xué)領(lǐng)域研究的熱點(diǎn)。BZG是一種化學(xué)結(jié)構(gòu)類似于索拉菲尼的多靶點(diǎn)酶抑制劑,我們既往的研究證實(shí),它可以在體外抑制腫瘤細(xì)胞的增殖,顯著抑制肝細(xì)胞肝癌異種移植模型中的腫瘤生長(zhǎng)。目的:本課題在我們前期研究工作的基礎(chǔ)上,擬應(yīng)用臨床前體內(nèi)模型,闡明BZG吸收、分布性質(zhì),明確其臨床前吸收代謝動(dòng)力學(xué)性質(zhì),為其臨床研究做好前期研究工作。本研究的最終目的是使BZG成為具有自主產(chǎn)權(quán)的一類新藥,為我國(guó)廣大肝癌患者提供新的藥物選擇,從而減輕患者治療所需的經(jīng)濟(jì)負(fù)擔(dān),進(jìn)而獲得良好的經(jīng)濟(jì)效益和社會(huì)效益。方法:本文采用UPLC-MS/MS在合適的色譜和質(zhì)譜條件下,利用伊馬替尼作為內(nèi)標(biāo),建立BZG在生物樣品中的測(cè)定方法。根據(jù)FDA生物分析方法驗(yàn)證指南驗(yàn)證其專屬性、線性、準(zhǔn)確度、精密度、回收率、基質(zhì)效應(yīng)和穩(wěn)定性。大鼠按照20mg/kg灌胃給藥后,在0至48小時(shí)內(nèi)的不同時(shí)間點(diǎn)取血液樣本,按照所建立的生物樣品中BZGUPLC-MS/MS測(cè)定法進(jìn)行測(cè)定,取得給藥后不同時(shí)間點(diǎn)的大鼠血漿中BZG的濃度,用DAS軟件分析獲得藥代動(dòng)力學(xué)參數(shù)。大鼠按照20mg/kg灌胃給藥后,1,6和12小時(shí)收集以下組織:小腸、肝臟、肺、腎臟、胰腺、肌肉、脂肪和大腦,按照所建立的生物樣品中BZGUPLC-MS/MS測(cè)定法進(jìn)行測(cè)定,取得給藥后不同時(shí)間點(diǎn)的大鼠不同組織中BZG的濃度,并據(jù)此研究藥物的組織分布。結(jié)果:本文成功建立BZG在生物樣品中的UPLC-MS/MS測(cè)定方法,其相關(guān)驗(yàn)證結(jié)果如下:在研究濃度范圍(0.5ng/ml-2500ng/ml)內(nèi),BZG具有良好的線性(r2≥0.994);BZG色譜圖中樣品峰無(wú)雜質(zhì)峰干擾,專屬性良好。低、中、高濃度的質(zhì)控樣品的日間、日內(nèi)準(zhǔn)確度為-11.65%至11.76%,精密度為0.34%至14.64%;回收率分別為 80.42%至 89.46%,RSD10.87%;基質(zhì)效應(yīng) 84.62%至 92.41%,RSD≤10.31%;常溫保存8h、-20℃保存40天、反復(fù)凍融3次以及處理后24h內(nèi)的BZG穩(wěn)定性在±15%之間。大鼠按照20mg/kg灌胃給藥后,用DAS軟件分析血藥濃度的主要結(jié)果如下:大鼠體內(nèi)的藥代動(dòng)力學(xué)行為符合一室模型;BZG在6.00±0.35小時(shí)達(dá)到587.08±84.08ng/ml的平均最大濃度;消除半衰期(t1/2)為2.86±2.49小時(shí)。大鼠按照20mg/kg灌胃給藥后,其重要臟器的組織分布主要結(jié)果如下:BZG能夠快速而廣泛地分布到大鼠各組織中;所有組織均在6小時(shí)達(dá)到最高濃度;小腸內(nèi)BZG濃度最高;然后依次是腎臟、肝臟、肺、胰腺、肌肉、脂肪和大腦。結(jié)論:1.本文建立并驗(yàn)證一種測(cè)定大鼠生物樣品中BZG濃度的UPLC-MS/MS方法。2.按照20mg/kg灌胃后,大鼠體內(nèi)的BZG在6.00±0.35小時(shí)達(dá)到587.08±84.08ng/ml的平均最大濃度;消除半衰期(t1/2)為2.86±2.49小時(shí),符合一室模型。3.按照20mg/kg灌胃后,BZG能夠快速而廣泛地分布到大鼠各組織中,組織濃度由高至低依次為小腸、腎臟、肝臟、肺、胰腺、肌肉、脂肪和大腦。
[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.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.7

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本文編號(hào)：2240364