Complex crystal structure determination and anti-non-small-cell lung cancer activity of the Hsp90N inhibitor Debio0932.
Complex crystal structure determination and anti-non-small-cell lung cancer activity of the Hsp90N inhibitor Debio0932.
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DOI：
10.1107/s2059798320014990
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发表时间：
2021-01
期刊：
影响因子：
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通讯作者：
Wei Qin;F. Yu;Huan Zhou;Ping Li;Fang Zhou;Hui Li;Chun-Xia He;Lu Xing;Xin Zhou;Dong Zhao;Pengquan Li;Xi Jin;Qi Wang;Jian-Hua He;H. Cao
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文献类型：
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作者：
Wei Qin;F. Yu;Huan Zhou;Ping Li;Fang Zhou;Hui Li;Chun-Xia He;Lu Xing;Xin Zhou;Dong Zhao;Pengquan Li;Xi Jin;Qi Wang;Jian-Hua He;H. Cao
Debio 0932是一个很有前途的先导化合物，在I期临床试验中靶向分子伴侣热休克蛋白90（Hsp 90 N）的N-末端ATP结合口袋。然而，Hsp 90 N-Debio 0932复合物的晶体结构的缺乏阻碍了Debio 0932的进一步结构优化和分子相互作用机制的理解。在这里，通过X射线衍射成功地确定了Hsp 90 N-Debio 0932复合物的高分辨率晶体结构（分辨率极限2.20 nm; PDB条目6lr 9），并详细分析了分子相互作用机制，这表明Debio 0932通过将其自身容纳在Hsp 90 N的ATP结合口袋中，使其分子伴侣能力丧失，从而抑制癌细胞。热位移测定（ΔTm = 8.83 ± 0.90°C）和等温滴定量热法（Kd = 15.50 ± 1.30 nM）的结果表明Hsp 90 N和Debio 0932的结合具有强结合和有利的热力学变化。基于复合物的晶体结构和分子相互作用分析，设计了30种新的Debio 0932衍生物，其中9种新衍生物表现出与Hsp 90 N的结合增加，通过分子对接评估确定。此外，Debio 0932抑制细胞增殖（A549的IC 50值为3.26 ± 2.82 µM，H1299为20.33 ± 5.39 µM，H1975为3.16 ± 1.04 µM），诱导细胞周期停滞并促进三种非小细胞肺癌（NSCLC）细胞系的凋亡。这些结果为基于先导化合物Debio 0932的新型抗NSCLC药物的开发提供了新的视角和指导。
Debio0932 is a promising lead compound in phase I clinical trials targeting the N-terminal ATP-binding pocket of the molecular chaperone heat-shock protein 90 (Hsp90N). The absence of a crystal structure of the Hsp90N-Debio0932 complex, however, has impeded further structural optimization of Debio0932 and understanding of the molecular-interaction mechanism. Here, a high-resolution crystal structure of the Hsp90N-Debio0932 complex was successfully determined (resolution limit 2.20 Å; PDB entry 6lr9) by X-ray diffraction and the molecular-interaction mechanism was analysed in detail, which suggested that Debio0932 suppresses cancer cells by accommodating itself in the ATP-binding pocket of Hsp90N, disabling its molecular-chaperone capability. The results of a thermal shift assay (ΔTm = 8.83 ± 0.90°C) and isothermal titration calorimetry (Kd = 15.50 ± 1.30 nM) indicated strong binding and favourable thermodynamic changes in the binding of Hsp90N and Debio0932. Based on the crystal structure of the complex and on molecular-interaction analysis, 30 new Debio0932 derivatives were designed and nine new derivatives exhibited increased binding to Hsp90N, as determined by molecular-docking evaluation. Additionally, Debio0932 suppressed cell proliferation (IC50 values of 3.26 ± 2.82 µM for A549, 20.33 ± 5.39 µM for H1299 and 3.16 ± 1.04 µM for H1975), induced cell-cycle arrest and promoted apoptosis in three non-small-cell lung cancer (NSCLC) cell lines. These results provide novel perspectives and guidance for the development of new anti-NSCLC drugs based on the lead compound Debio0932.