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MLN0905(6,8,10,6.5和14.5mg\u002Fkg,po)在原发性人淋巴瘤模型(PHTX-22L)中引起显着的抗肿瘤反应。此外,MLN0905在播散性OCI LY-19-Luc淋巴瘤模型中与利妥昔单抗协同作用[2]。",{"key":245,"value":246},"激酶实验","总共30μL的人PLK1酶促反应含有50mM Tris-HCl（pH 8.0），10mM MgCl 2,0.02％BSA，10％甘油，1mM DTT，100mM NaCl，3.3％DMSO，50μMATP，2μM肽底物（生物素-AHX-LDETGHLDSSGLQEVHLA-CONH2）和0.3nM重组人PLK1 [2-369] T210D。将含有或不含PLK抑制剂的酶促反应混合物在室温下孵育90分钟，然后用50μL含有1％BSA，0.05％吐温20,100mM EDTA的STOP缓冲液终止。然后将50μL终止的酶反应混合物转移到涂有Neutravidin的384孔板中，并在室温下温育60分钟。用洗涤缓冲液（25mM Tris，150mM氯化钠和0.1％Tween 20）洗涤孔，并与含有1％BSA，0.05％Tween 20，抗磷酸化cdc25c兔单克隆的50μL抗体反应混合物一起温育1小时。抗体（325pM）和铕标记的抗兔IgG（2nM）。洗涤孔，然后使用50μL增强溶液释放结合的铕。使用Pherastar [1]进行铕的定量分析。",{"key":248,"value":249},"细胞实验","将8μL连续稀释的测试化合物加入到McCoy's 5A培养基中的75μLHT29（2.66×104细胞\u002F孔）细胞中，所述培养基补充有Biocoat Poly-D赖氨酸384孔黑\u002F透明板中的10％胎牛血清。将细胞在37℃下孵育72小时。从孔中吸出上清液，在每个孔中留下25μL。向每个孔中加入ATP-lite 1步试剂（25μL），并在LeadSeeker上读取每个板的发光。使用DMSO对照组的值至100％[1]计算抑制百分比。",{"key":251,"value":252},"动物实验","HT29细胞从ATCC获得，并在补充有10％FBS的McCoy's 5A培养基中培养。将HT29细胞（2×10 6）重悬于Hanks缓冲液中并皮下注射到雌性裸鼠的侧腹中。小鼠（10只动物\u002F组）用12c（MLN0905）口服治疗21天，剂量基于耐受性结果：QD（6.25和12.5mg \u002F kg），QD×1 \u002F周（50mg \u002F kg）和QD× 3 \u002F周（25毫克\u002F千克）。使用游标卡尺监测肿瘤生长，并使用公式V = W2×L \u002F 2计算平均肿瘤体积。当平均肿瘤体积达到约200mm 3时，将动物随机分成治疗组（n = 10只动物\u002F组）。每周测量肿瘤生长和体重两次[1]。",{"key":254,"value":255},"参考文献","[1].\nDuffey MO, et al. 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