Identification of a potent Nrf2 displacement activator among aspirin-containing prodrugs.
Identification of a potent Nrf2 displacement activator among aspirin-containing prodrugs.
复制标题
DOI：
10.1016/j.neuint.2021.105148
复制
发表时间：
2021-10
影响因子：
4.2
通讯作者：
Thomas B
中科院分区：
文献类型：
--
作者：
Gaisina IN;Hushpulian DM;Gaisin AM;Kazakov EH;Ammal Kaidery N;Ahuja M;Poloznikov AA;Gazaryan IG;Thatcher GRJ;Thomas B
阿司匹林是治疗神经退行性疾病和其他疾病的前药中理想的离去基团。在Neh2-luc报告基因试验中，研究人员对可能激活Nrf2的各种支架的阿司匹林衍生物库进行了测试，该试验筛选通过破坏Nrf2- keap1相互作用直接起作用的Nrf2蛋白稳定剂。大多数阿司匹林前药在结构中含有前烷基化基序或前氧化基序，因此在高浓度时是有毒的。然而，在活性化合物中，我们发现了一种类似于Nrf2位移激活剂的分子，即双-1,4-（4-甲氧基苯磺酰胺基）萘（NMBSA）。新鉴定的化合物与NMBSA及其改进的类似物在报告实验中直接比较，发现n -乙酰半胱氨酸不猝灭，从而指出Nrf2的稳定机制不需要半胱氨酸烷基化。尽管在商业荧光偏振分析中仅在毫摩尔范围内观察到其抑制作用，但新鉴定的化合物在报告细胞实验中的效力比NMBSA强得多。分子对接预测，新型前药的单去乙酰化应该产生一个有效的位移激活剂。新的前药的报告细胞激活的时间过程有一个明显的滞后期，指向一个可能的细胞内转化导致活性产物。通过液相色谱-质谱（LC-MS）分析，血浆或细胞裂解液对新型前药的处理表现为逐步去乙酰化。因此，酯酶催化的前药水解仅从阿司匹林片段中释放乙酰基，并产生有效的Nrf2激活剂。发现的前药活化机制使新鉴定的化合物成为未来优化研究的有希望的先导物。
Aspirin is a desired leaving group in prodrugs aimed at treatment of neurodegeneration and other conditions. A library of aspirin derivatives of various scaffolds potentially activating Nrf2 has been tested in Neh2-luc reporter assay which screens for direct Nrf2 protein stabilizers working via disruption of Nrf2-Keap1 interaction. Most aspirin prodrugs had a pro-alkylating or pro-oxidant motif in the structure and, therefore, were toxic at high concentrations. However, among the active compounds, we identified a molecule resembling a well-known Nrf2 displacement activator, bis-1,4-(4-methoxybenzenesulfonamidyl) naphthalene (NMBSA). The direct comparison of the newly identified compound with NMBSA and its improved analog in the reporter assay showed no quenching with N-acetyl cysteine, thus pointing to Nrf2 stabilization mechanism without cysteine alkylation. The potency of the newly identified compound in the reporter assay was much stronger than NMBSA, despite its inhibitory action in the commercial fluorescence polarization assay was observed only in the millimolar range. Molecular docking predicted that mono-deacetylation of the novel prodrug should generate a potent displacement activator. The time-course of reporter activation with the novel prodrug had a pronounced lag-period pointing to a plausible intracellular transformation leading to an active product. Treatment of the novel prodrug with blood plasma or cell lysate demonstrated stepwise deacetylation as judge by liquid chromatography-mass spectrometry (LC-MS). Hence, the esterase-catalyzed hydrolysis of the prodrug liberates only acetyl groups from aspirin moiety and generates a potent Nrf2 activator. The discovered mechanism of prodrug activation makes the newly identified compound a promising lead for future optimization studies.
登录
查看更多内容
影响因子：
3.8
作者：
Kastrati I;Litosh VA;Zhao S;Alvarez M;Thatcher GR;Frasor J
通讯作者：
Frasor J
针对KEAP1-NRF2途径的挑战和局限性神经疗法：Bach1将抑制降低到营救。
影响因子：
4.8
作者：
Hushpulian DM;Ammal Kaidery N;Ahuja M;Poloznikov AA;Sharma SM;Gazaryan IG;Thomas B
通讯作者：
Thomas B
影响因子：
4.1
作者：
Dunlap T;Piyankarage SC;Wijewickrama GT;Abdul-Hay S;Vanni M;Litosh V;Luo J;Thatcher GR
通讯作者：
Thatcher GR
影响因子：
4.1
作者：
Dunlap, Tareisha;Chandrasena, R. Esala P.;Thatcher, Gregory R. J.
通讯作者：
Thatcher, Gregory R. J.
影响因子：
4.2
作者：
Lazzara, Phillip R.;Jain, Atul D.;Moore, Terry W.
通讯作者：
Moore, Terry W.