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(IC50)\nTANK-1:570 nM (IC50)\nPARP-3:1300 nM (IC50)",{"key":241,"value":242},"体外研究","在全细胞试验中,Niraparib(MK-4827)抑制PARP活性,EC50 = 4nM,EC90 = 45nM。 MK-4827用突变体BRCA-1和BRCA-2抑制癌细胞的增殖,CC50在10-100nM范围内。 MK-4827显示出优异的PARP1和2抑制,IC50分别为3.8和2.1nM,并且在全细胞试验中[1]。为了验证Niraparib(MK-4827)抑制这些细胞系中的PARP,用1μMMK-4827处理A549和H1299细胞不同时间并使用化学发光测定法测量PARP酶活性。结果显示,Niraparib(MK-4827)在处理后15分钟内抑制PARP,在1小时时A549细胞抑制约85％,H1299细胞在1小时抑制约55％[2]。",{"key":244,"value":245},"体内研究","Niraparib(MK-4827)具有良好的耐受性,并且在BRCA-1缺陷型癌症的异种移植模型中显示出作为单一药剂的功效。 Niraparib(MK-4827)在体内具有良好的耐受性,并且在BRCA-1缺陷型癌症的异种移植模型中显示出作为单一药剂的功效。 Niraparib(MK-4827)的特点是大鼠的药代动力学可接受,血浆清除率为28(mL\u002Fmin)\u002F kg,分布容积非常高(Vdss = 6.9 L\u002Fkg),终末半衰期长(t1\u002F2 = 3.4 h),和优异的生物利用度,F = 65％[1]。在两种情况下,Niraparib(MK-4827)均可增强p53突变Calu-6肿瘤的放射反应,每天两次给药的单剂量50 mg\u002Fkg比25 mg\u002Fkg更有效[3]。",{"key":247,"value":248},"细胞实验","使用HT Universal Chemiluminescent PARP Assay Kit在A549和H1299细胞中分析PARP的抑制。简言之，将细胞用DMSO或1μM镍吡啶（MK-4827）处理15,30,60或120分钟，胰蛋白酶消化，并转移至预冷管中。用冰冷的PBS洗涤细胞两次，并重悬于冷的PARP提取缓冲液中。将细胞悬浮液在冰上孵育30分钟，周期性涡旋以破坏细胞膜。离心悬浮液，将上清液转移到冰上预冷的管中。将96孔板的组蛋白包被孔用1X PARP缓冲液再水化，并在室温下孵育30分钟。除去PARP缓冲液，并将通过Bio-Rad蛋白质测定法测定的20μg蛋白质加入每个孔中，然后加入稀释的PARP-HSA酶和1X PARP缓冲液。然后将条带孔在室温下孵育60分钟，用含有0.1％Triton X-100的PBS洗涤两次，然后用PBS洗涤。将稀释的Strep-HRP加入到条带孔中并在室温下孵育60分钟。如前所述再次洗涤孔。将等体积的PeroxyGlow A和B组合并添加到孔中，并使用读板器立即获得化学发光读数[2]。",{"key":250,"value":251},"动物实验","小鼠[3]将雌性裸鼠（Ncr Nu \u002F Nu）随机分配到治疗组，每组由5至8只小鼠组成，当肿瘤直径增大至6.0mm时，此时开始用Niraparib（MK-4827）治疗。 Niraparib（MK-4827）以25mg \u002F kg的剂量每日两次或50mg \u002F kg每天一次给药21天，或者在肿瘤直径达到8mm的第9天停药。当肿瘤直径达到8.0mm（7.7-8.2mm）时，给予分次局部肿瘤照射（XRT）。使用由两个平行相对的137Cs源组成的小动物辐照器，每天一次连续14天或每天两次将辐射（每个部分2Gy）递送至小鼠的肿瘤腿部连续14天或连续7天，剂量率为5戈瑞\u002F分钟。在照射期间，将未麻醉的小鼠机械固定在夹具中，使得肿瘤在3.0cm直径的辐射场内居中，并且动物的身体避免辐射暴露。在给予Niraparib和放射的那天，在当天的第一次辐射剂量之前1小时施用药物。",{"key":253,"value":254},"参考文献","[1].\nJones P, et al. 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