Benzodiazepinedione inhibitors of the Hdm2:p53 complex suppress human tumor cell proliferation in vitro and sensitize tumors to doxorubicin in vivo
Benzodiazepinedione inhibitors of the Hdm2:p53 complex suppress human tumor cell proliferation in vitro and sensitize tumors to doxorubicin in vivo
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DOI：
10.1158/1535-7163.mct-05-0199
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发表时间：
2006-01-01
影响因子：
5.7
通讯作者：
Maroney, AC
中科院分区：
文献类型：
--
作者：
Koblish, HK;Zhao, SY;Maroney, AC
p53肿瘤抑制因子的活性和稳定性受小鼠双微体2（Hdm 2）癌蛋白的人类同源物调节。据推测，破坏Hdm 2：p53复合物的小分子将允许p53的活化并导致生长抑制。我们已经确定了小分子抑制剂的HDM 2：p53相互作用，使用我们专有的ThermoFluor微量热技术。药物化学和基于结构的药物设计导致了一系列优化的苯二氮卓二酮类药物的开发，包括TDP 521252和TDP 665759。活性依赖于野生型（wt）p53和Hdm 2的表达，如通过在突变或无效p53表达细胞系或经工程改造不再表达Hdm 2和wt p53的细胞中缺乏效力所确定的。TDP 521252和TDP 665759抑制表达wt p53的细胞系的增殖，平均IC 50分别为14和0.7 μ mol/L。这些结果与在绒毛膜癌细胞中15分钟内观察到的Hdm 2与wt p53的直接细胞解离相关。这些抑制剂在体外的其他活性包括稳定p53蛋白水平，以DNA损伤非依赖性方式上调p53靶基因，以及诱导HepG 2细胞凋亡。对小鼠给予TDP 665759导致肝脏样品中p21（waf 1/cip 1）水平升高。最后，在培养物和A375异种移植模型中，TDP 665759与多柔比星协同作用以减少肿瘤生长。总之，这些数据支持Hdm 2：p53相互作用的小分子抑制剂用于治疗表达野生型p53的肿瘤的潜在效用。
The activity and stability of the p53 tumor suppressor are regulated by the human homologue of the mouse double minute 2 (Hdm2) oncoprotein. It has been hypothesized that small molecules disrupting the Hdm2:p53 complex would allow for the activation of p53 and result in growth suppression. We have identified small-molecule inhibitors of the Hdm2:p53 interaction using our proprietary ThermoFluor microcalorimetry technology. Medicinal chemistry and structure-based drug design led to the development of an optimized series of benzodiazepinediones, including TDP521252 and TDP665759. Activities were dependent on the expression of wild-type (wt) p53 and Hdm2 as determined by lack of potency in mutant or null p53-expressing cell lines or cells engineered to no longer express Hdm2 and wt p53. TDP521252 and TDP665759 inhibited the proliferation of wt p53-expressing cell lines with average IC50S Of 14 and 0.7 mu mol/L, respectively. These results correlated with the direct cellular dissociation of Hdm2 from wt p53 observed within 15 minutes in JAR choriocarcinoma cells. Additional activities of these inhibitors in vitro include stabilization of p53 protein levels, up-regulation of p53 target genes in a DNA damage-independent manner, and induction of apoptosis in HepG2 cells. Administration of TDP665759 to mice led to an increase in p21(waf1/cip1) levels in liver samples. Finally, TDP665759 synergizes with doxorubicin both in culture and in an A375 xenograft model to decrease tumor growth. Taken together, these data support the potential utility of small-molecule inhibitors of the Hdm2:p53 interaction for the treatment of wt p53-expressing tumors.