Laboratório de Biologia Molecular, Centro Infantil Boldrini, Campinas, SP, 13083-210, Brazil; Graduate Program in Genetics and Molecular Biology, State University of Campinas, Campinas, SP, 13083-210, Brazil.
Institute of Chemistry, State University of Campinas, Campinas, SP, 13083-970, Brazil.
Eur J Med Chem. 2019 Nov 1;181:111570. doi: 10.1016/j.ejmech.2019.111570. Epub 2019 Jul 31.
Despite the success achieved in the treatment of acute lymphoblastic leukemia (ALL), the search for new drugs featuring selectivity against leukemia cells and effectiveness to prevent relapsed ALL is still highly desirable. Here, we described the synthesis of several novel 3-substituted and 3,6-disubstituted-2-carboalkoxy indoles followed by the elucidation of their mechanism of action and in vivo anti-leukemia efficacy. The synthesis of 3-substituted-2-carboalkoxy indoles relied on two Heck arylations of methyl acrylate and methyl cinnamates respectively, to generate β,β-disubstituted acrylates followed by an efficient Cadogan-Sundberg reaction of these latter intermediates. The method developed led to the synthesis of twenty-one novel functionalized indoles. Of these, indole 20 showed selective cytotoxicity against leukemia cells at the nanomolar scale, and, therefore, it was selected for the investigation of its mechanism of action. Indole 20 was found to target tubulin leading to G2/M cell cycle arrest, DNA damage and apoptosis. Indole 20 decreased β-tubulin protein in leukemia cells in a time-dependent manner and induced depolymerization of the microtubule network in Hela cells, thus fully characterizing its microtubule destabilizer activity. The connectivity map analysis of HL60 promyelocytic leukemia cells treated with indole 20 revealed a transcriptional profile similar to that of cells treated with prostaglandins, apparently due to the induction of cellular differentiation as addressed by the expression of CD11 and CD14 markers. Finally, indole 20 given intraperitoneally, at 10 mg/kg, 5x/week significantly prolonged the overall survival of NOD/SCID mice transplanted with RS4; 11 B-ALL cells.
尽管在治疗急性淋巴细胞白血病 (ALL) 方面取得了成功，但仍非常需要寻找具有选择性针对白血病细胞的疗效并能预防 ALL 复发的新药。在这里，我们描述了几种新型 3-取代和 3,6-二取代-2-羧酸吲哚的合成，随后阐明了它们的作用机制和体内抗白血病功效。3-取代-2-羧酸吲哚的合成依赖于甲基丙烯酸酯和肉桂酸甲酯的两次 Heck 芳基化反应，分别生成β,β-二取代丙烯酰胺，然后通过这些中间体的 Cadogan-Sundberg 反应高效合成。所开发的方法导致了二十一种新型功能化吲哚的合成。其中，吲哚 20 在纳摩尔范围内对白血病细胞具有选择性细胞毒性，因此被选为其作用机制的研究对象。吲哚 20 被发现靶向微管蛋白导致 G2/M 细胞周期停滞、DNA 损伤和细胞凋亡。吲哚 20 以时间依赖性方式降低白血病细胞中的β-微管蛋白蛋白，并诱导 Hela 细胞中的微管网络解聚，从而充分表征其微管不稳定活性。用吲哚 20 处理 HL60 早幼粒细胞白血病细胞的连接图谱分析显示出与用前列腺素处理的细胞相似的转录谱，显然是由于细胞分化的诱导，如 CD11 和 CD14 标志物的表达所表明的那样。最后，在 RS4 ；11B-ALL 细胞移植的 NOD/SCID 小鼠中，以 10mg/kg、每周 5 次腹腔内给予吲哚 20 可显著延长总体存活时间。
