Magalhães Emanuel Paula, Gomes Naiara Dutra Barroso, Freitas Tiago Araújo de, Silva Brenna Pinheiro, Ribeiro Lyanna Rodrigues, Ameida-Neto Francisco Wagner Queiroz, Marinho Márcia Machado, Lima-Neto Pedro de, Marinho Emmanuel Silva, Santos Hélcio Silva Dos, Teixeira Alexandre Magno Rodrigues, Sampaio Tiago Lima, Menezes Ramon Róseo Paula Pessoa Bezerra de, Martins Alice Maria Costa
Post-Graduate Program in Pharmaceutical Sciences, Federal University of Ceará, Fortaleza, CE, Brazil.
Department of Analytical Chemistry and Physical Chemistry, Science Center, Federal University of Ceará, Campus do Pici, Fortaleza, CE, Brazil.
Chem Biol Interact. 2022 Jul 1;361:109920. doi: 10.1016/j.cbi.2022.109920. Epub 2022 Apr 21.
Chagas disease is a disease that is emerging in North America and Europe countries. Benznidazole is the main drug available, but it has high toxicity and low efficacy in the chronic phase. In this way, researching new antichagasic agents is necessary. Thus, the aim of this study is to evaluate the effect of novel chalcones and the influence of chlorine substitutions on Trypanosoma cruzi and host cells. Unsubstituted (1), 4-chlorine substituted (2) and 2,4-chlorine substituted (3) chalcones were synthesized by Claisen-Schmidt condensation, characterized, and electrical distribution was assessed by Density Fuctional Theory (DFT). The host cells toxicity (LLC-MK2) was performed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) reduction assay. The effect on epimastigote (24, 48 and 72h), trypomastigote (24h) and amastigotes (24 h) was evaluated. Flow cytometry assays were performed with 7-Aminoactinomycin D (7-AAD) and Annexin-PE, Dichlorofluorescein diaceteate (DCFH-DA) and Rhodamine123 (Rho123). Finally, molecular docking predicted interactions between chalcones and cruzain (TcCr) and trypanothione reductase (TcTR). The toxicity on host cells was reduced almost twenty times on chlorine substituted molecules. On epimastigote and trypomastigote forms, all substances presented similar effects. After treatment with molecule 3, it was observed a decrease in infected cells and intracellular amastigotes. Their effect is related to necrotic events, increase of cytoplasmic Reactive Oxygen Species (ROS) and mitochondrial dysfunction. Also, this effect might be associated with involvement of TcCr and TcTR enzymes. Therefore, the results showed that chlorine substitution on chalcones reduces the host cell's toxicity without compromising the effect on Trypanosoma cruzi Y strain forms, and it occurs over membrane damage, oxidative stress and possible interactions with TcCr and TcTR.
恰加斯病正在北美和欧洲国家出现。苯硝唑是现有的主要药物，但它在慢性期毒性高、疗效低。因此，研究新的抗锥虫药物很有必要。本研究的目的是评估新型查耳酮的作用以及氯取代对克氏锥虫和宿主细胞的影响。通过克莱森-施密特缩合反应合成了未取代的（1）、4-氯取代的（2）和2,4-氯取代的（3）查耳酮，对其进行了表征，并通过密度泛函理论（DFT）评估了电子分布。通过3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐（MTT）还原试验检测宿主细胞毒性（LLC-MK2）。评估了对前鞭毛体（24、48和72小时）、锥鞭毛体（24小时）和无鞭毛体（24小时）的影响。使用7-氨基放线菌素D（7-AAD）和膜联蛋白-PE、二氯荧光素二乙酸酯（DCFH-DA）和罗丹明123（Rho123）进行流式细胞术检测。最后，分子对接预测了查耳酮与克鲁兹蛋白酶（TcCr）和锥虫硫醇还原酶（TcTR）之间的相互作用。氯取代分子对宿主细胞的毒性降低了近20倍。在前鞭毛体和锥鞭毛体形式上，所有物质都表现出相似的效果。用分子3处理后，观察到感染细胞和细胞内无鞭毛体减少。它们的作用与坏死事件、细胞质活性氧（ROS）增加和线粒体功能障碍有关。此外，这种作用可能与TcCr和TcTR酶的参与有关。因此，结果表明，查耳酮上的氯取代降低了宿主细胞的毒性，同时不影响对克氏锥虫Y株形式的作用，其作用通过膜损伤、氧化应激以及与TcCr和TcTR的可能相互作用而发生。
