Santos Bubniak Lorena Dos, Gaspar Pâmela Cristina, de Moraes Ana Carolina Rabello, Bigolin Alisson, de Souza Rubia Karine, Buzzi Fátima Campos, Corrêa Rogério, Filho Valdir Cechinel, Bretanha Lizandra Czermainski, Micke Gustavo Amadeu, Nunes Ricardo José, Santos-Silva Maria Cláudia
a Departamento de Análises Clínicas, Universidade Federal de Santa Catarina - UFSC, 89, Campus Trindade, CEP 88040-900, Florianópolis, Santa Catarina, Brazil.
c Núcleo de Investigações Químico-Farmacêuticas (NIQFAR), Universidade do Vale de Itajaí, UNIVALI, CEP - Itajaí, Santa Catarina, Brazil.
Can J Physiol Pharmacol. 2017 May;95(5):548-563. doi: 10.1139/cjpp-2016-0222. Epub 2016 Nov 24.
Pyrazoline is an important 5-membered nitrogen heterocycle that has been extensively researched. Ten derivatives were synthesized and tested for antileukemic effects on 2 human acute leukemia cell lines, K562 and Jurkat. The most cytotoxic of these derivatives, compound 21, was chosen for investigation of cytotoxicity mechanisms. The results obtained with selectivity calculations revealed that compound 21 is more selective for acute leukemia (K562 and Jurkat cell lines) than for other tumor cell lines. Moreover, compound 21 was not cytotoxic to normal cell lines, indicating a potential use in clinical tests. Compound 21 caused a significant cell cycle arrest in the S-phase in Jurkat cells and increased the proportion of cells in the sub G0/G1 phase in both cell lines. Cells treated with compound 21 demonstrated morphological changes characteristic of apoptosis in the EB/AO assay, confirmed by externalization of phosphatidylserine by the annexin V - fluorescein isothiocyanate method and by DNA fragmentation. An investigation of cytotoxicity mechanisms suggests the involvement of an intrinsic apoptosis pathway due to mitochondrial damage and an increase in the ratio of mitochondrial Bax/Bcl2. Pyrazoline 21 obeyed Lipinski's "rule of five" for drug-likeness. Based on these preliminary results, the antileukemic activity of compound 21 makes it a potential anticancer agent.
吡唑啉是一种重要的五元氮杂环，已被广泛研究。合成了十种衍生物，并对其对两种人类急性白血病细胞系K562和Jurkat的抗白血病作用进行了测试。选择这些衍生物中细胞毒性最强的化合物21来研究其细胞毒性机制。选择性计算结果表明，化合物21对急性白血病（K562和Jurkat细胞系）的选择性高于其他肿瘤细胞系。此外，化合物21对正常细胞系无细胞毒性，表明其在临床试验中有潜在应用价值。化合物21使Jurkat细胞在S期发生显著的细胞周期阻滞，并增加了两种细胞系中亚G0/G1期细胞的比例。用化合物21处理的细胞在EB/AO试验中表现出凋亡的形态学变化，通过膜联蛋白V - 异硫氰酸荧光素法检测磷脂酰丝氨酸外翻以及DNA片段化得到证实。细胞毒性机制研究表明，由于线粒体损伤以及线粒体Bax/Bcl2比值增加，存在内源性凋亡途径。吡唑啉21符合Lipinski的药物相似性“五规则”。基于这些初步结果，化合物21的抗白血病活性使其成为一种潜在的抗癌药物。
