通过小分子筛选鉴定抗酒石酸酸性磷酸酶（TRAP / ACP5）活性的抑制剂,Chemical Biology & Drug Design

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通过小分子筛选鉴定抗酒石酸酸性磷酸酶（TRAP / ACP5）活性的抑制剂
Chemical Biology & Drug Design ( IF 3.2 ) Pub Date : 2018-03-30 , DOI: 10.1111/cbdd.13187
Anja Reithmeier ₁ , Thomas Lundbäck ₂ , Martin Haraldsson ₂ , Martin Frank ₃ , Barbro Ek-Rylander ₁ , Per-Georg Nyholm ₃ , Anna-Lena Gustavsson ₂ , Göran Andersson ₁

Affiliation

抗酒石酸酸性磷酸酶（TRAP / ACP5）以两种同工型出现-具有低酶促活性的TRAP 5a，这是由于一个环与活性位点相互作用，以及通过蛋白水解切割该环而产生的更具活性的TRAP同工型5b。TRAP与多种疾病有关，包括癌症。因此，本研究着手确定TRAP活性的小分子抑制剂。在针对30,315种化合物的筛选中进行了基于微孔板的TRAP 5b酶促测定，鉴定出90种主要命中物。通过正交试验和三浓度验证去除了混杂化合物，不需要的基团和假阳性后，进一步研究了52种化合物的性质，以更好地了解其作用机理。在不同的酶浓度和（预）温育时间下建立这些化合物的全浓度-响应曲线，以去除结果不一致且效力低的化合物。还对两种同工型都绘制了全浓度-响应曲线，以检查同工型的患病率。筛选导致六个优先化合物的出现，代表了不同的簇。其中之一是CBK289001或（6S）‐6‐ [3‐（2H‐1,3-‐苯并二恶酚‐5‐基）‐1,2,4-恶二唑‐5‐基] ‐N‐（丙-2-基）-1H，4H，5H，6H，7H-咪唑并[4,5-c]吡啶-5-羧酰胺，在迁移分析和IC中显示出功效代表不同的集群。其中之一是CBK289001或（6S）‐6‐ [3‐（2H‐1,3-‐苯并二恶酚‐5‐基）‐1,2,4-恶二唑‐5‐基] ‐N‐（丙-2-基）-1H，4H，5H，6H，7H-咪唑并[4,5-c]吡啶-5-羧酰胺，在迁移分析和IC中显示出功效代表不同的集群。其中之一是CBK289001或（6S）‐6‐ [3‐（2H‐1,3-‐苯并二恶酚‐5‐基）‐1,2,4-恶二唑‐5‐基] ‐N‐（丙-2-基）-1H，4H，5H，6H，7H-咪唑并[4,5-c]吡啶-5-羧酰胺，在迁移分析和IC中显示出功效_50个4至125μ值米。围绕CBK289001进行了分子对接研究和模拟测试，为进一步改进TRAP活性的更强阻断剂提供了机会。

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Identification of inhibitors of Tartrate‐resistant acid phosphatase (TRAP/ACP5) activity by small‐molecule screening

Tartrate‐resistant acid phosphatase (TRAP/ACP5) occurs as two isoforms—TRAP 5a with low enzymatic activity due to a loop interacting with the active site and the more active TRAP isoform 5b generated upon proteolytic cleavage of this loop. TRAP has been implicated in several diseases, including cancer. Thus, this study set out to identify small‐molecule inhibitors of TRAP activity. A microplate‐based enzymatic assay for TRAP 5b was applied in a screen of 30,315 compounds, resulting in the identification of 90 primary hits. After removal of promiscuous compounds, unwanted groups, and false positives by orthogonal assays and three‐concentration validation, the properties of 52 compounds were further investigated to better understand their mechanism of action. Full‐concentration–response curves for these compounds were established under different enzyme concentrations and (pre)incubation times to remove compounds with inconsistent results and low potencies. Full‐concentration–response curves were also performed for both isoforms, to examine isoform prevalence. Filtering led to six prioritized compounds, representing different clusters. One of these, CBK289001 or (6S)‐6‐[3‐(2H‐1,3‐benzodioxol‐5‐yl)‐1,2,4‐oxadiazol‐5‐yl]‐N‐(propan‐2‐yl)‐1H,4H,5H,6H,7H‐imidazo[4,5‐c]pyridine‐5‐carboxamide, demonstrated efficacy in a migration assay and IC₅₀ values from 4 to 125 μm. Molecular docking studies and analog testing were performed around CBK289001 to provide openings for further improvement toward more potent blockers of TRAP activity.

更新日期：2018-03-30

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