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plus和TRI-CARB通过液体闪烁计数进行分析。酶活性表示为每毫克总细胞蛋白转移至TCA不溶性蛋白质材料的[3H]甲基的fmol。相对于未处理的对照样品计算抑制百分比。每个测定重复三次，并且从抑制百分比对抑制剂浓度的图中以图形方式确定IC 50值[1]。",{"key":260,"value":261},"细胞实验","为了确定毒性，使用MTT生长抑制测定法。将细胞（每孔1000个）接种到96孔板中，并在24小时附着期后，将Lomeguatrib加入细胞中。与Lomeguatrib（10μM）在37℃，5％CO 2下孵育2小时后，加入增加剂量的替莫唑胺或载体，并将细胞再孵育4-5天。在暴露期结束时，向每个孔中加入150μgMTT，并将板在37℃，5％CO 2下孵育3小时。除去培养基，将活细胞中形成的甲crystals晶体溶解在200μLDMSO中。使用ELISA板读数器测定540和690nm处的吸光度，并计算生长抑制作为未处理孔的A540-A690的百分比[2]。",{"key":263,"value":264},"动物实验","小鼠[2]为了评估Lomeguatrib使人乳腺肿瘤异种移植物对替莫唑胺的肿瘤生长抑制作用敏感的能力，对至少6只裸鼠的组进行如下处理：载体对照组给予玉米油，然后给予20％DMSO。 PBS;仅替莫唑胺组给予玉米油，然后给予替莫唑胺（100mg \u002F kg \u002F天）;仅Lomeguatrib组给予Lomeguatrib（20mg \u002F kg \u002F天），然后给予PBS中的DMSO，Lomeguatrib加替莫唑胺组给予Lomeguatrib（20mg \u002F kg \u002F天），然后给予替莫唑胺（100mg \u002F kg \u002F天）。每天一次腹膜内施用药物或载体，持续5天，间隔1小时。每组最多分配10只动物和至少6只动物，并且在实验开始时各组的平均肿瘤体积是标准化的：因此对照组，Lomeguatrib，替莫唑胺和Lomeguatrib加替莫唑胺组的平均肿瘤体积为29.8±7.6。 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Monosaccharide-linked inhibitors of O(6)-methylguanine-DNA methyltransferase (MGMT): synthesis, molecular modeling, and structure-activity relationships. J Med Chem. 2001 Nov 22;44(24):4050-61.\n[2].\nClemons M, et al. O6-(4-bromothenyl)guanine reverses temozolomide resistance in human breast tumour MCF-7 cells and xenografts. 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