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摘要：Bcr-abl(breakpoint cluster region Abelson)是一种异常的融合蛋白，与费城染色体(Philadelphia chromosome,Ph)阳性白血病紧密关联。Bcr-abl融合基因表达出的酪氨酸激酶能引起细胞的增殖、黏附和生存性质的变化，可导致多种肿瘤的产生。作为全球十大恶性肿瘤之一，白血病严重危害到人们的身体健康。因此Bcr-abl 是治疗Ph+白血病比较理想的靶点药物。本论文旨在建立测定Bcr-abl的有关物质。采用高效液相色谱法。色谱柱选用Agilent 5 TC-C18(2) 250×4.6mm。流动相A=在1000ml水中加1.0ml三氟乙酸，用三乙胺调节PH=3.0。流动相B=乙腈。采用梯度洗脱，流速控制在1.0ml•min–1，检测波长为252nm，柱温为35℃。根据专属性试验结果初步判断可能存在4个杂质。经过一系列验证（包括线性&范围，精密度，耐用性，检测限，定量限和准确性），本法操作简便、快速，结果准确可靠，可用于Bcr-abl有关物质的测定。21421
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毕业论文关键词：高效液相色谱法，Bcr-abl，有关物质，专属性，方法验证
The research of quality standard and the process of making the quality standard for a class of new drugs Bcr-abl
Abstract: Bcr-abl(breakpoint cluster region Abelson) is an abnormal fusion protein which is closely related with Ph(Philadelphia chromosome) positive leukemia. The tyrosine kinases expressed by Bcr-abl fusion gene can cause cell proliferation, adhesion and survival natural change, several kinds of tumors can be caused by it. As one of the world’s top ten malignant tumors, 1eukemia seriously endangers people’s health. Therefore, Bcr-abl is an ideal target for the treatment of Ph+ leukemia. This paper establishes the method for the content determination of main component in Bcr-abl. HPLC method was adopted. The separation was achieved by Agilent 5 TC-C18(2) 250×4.6mm column. The mobile phase A is prepared by dissolving 1.0ml trifluoroacetic acid dissolved in 1000mL of water, and pH adjusted to 3.5 with triethylamine. Mobile phase B is acetonitrile. The mobile phase B is acetonitrile. The mobile phase uses gradient elution. The flow rate of the mobile phase is maintained 1.0 mL•min–1. The detection wavelength was set at 252 nm and the column temperature was 35℃. According to the specificity test, four potential impurities finded preliminary. The developed HPLC method is validated with respect to linearity&range, precision, durability, LOD, LOQ and accuracy for impurities determination and assay determination. The method is simple, rapid and reproducible, and it is suitable for the content determination in Bcr-abl.
Keywords：HPLC, Bcr-abl, Content determination, Specificity, Validation
目录
1 前言 1
1.1 药物研究背景 1
1.2 药物发展历史 2
1.3 药物介绍 3
1.4 药物研究的难点 3
2 实验部分 4
2.1 仪器与试药 4
2.1.1 仪器 4
2.1.2 试药 4
2.2 分析方法的建立 4
2.2.1 固定相的筛选 4
2.2.2 溶剂的筛选 5
2.2.3 流动相筛选 5
2.2.4 流动相PH筛选 6
2.2.5 梯度筛选 7
2.2.6 柱温筛选 8
2.2.7 波长筛选 9
2.2.8 校正因子计算 10
2.3 专属性试验 11
2.3.1 强制破坏 11
2.3.2 物料平衡-强制降解试验检测到的物料平衡分析 12
2.3.3 杂质研究 12
2.4 方法学验证 17
2.4.1 线性试验 17
2.4.2 精密度试验 21
2.4.3 耐用性 22