Abstract

Chromene and its derivatives are generally spread in nature. Heterocylic-based compounds like chromenes have displayed pharmacological activities. Chromene derivatives are critical due to some biological features such as anticancer activity. CML, chronic myelogenous leukemia, is a fatal malignancy determined by resistance to apoptosis and contains the Philadelphia chromosome. Induction of apoptosis is one of the main approaches in cancer therapy. In this research, benzochromene derivative, 2-amino-4-(4-methoxy phenyl)-4H-benzochromene-3-carbonitrile (4-MC) was tested for cytotoxic and apoptotic induction activities in the human leukemic K562 cell line. The MTT growth inhibition assay was used to determine the cellular growth and survival. Moreover, the binding attribute of 4-MC with double helix DNA was assessed by some spectroscopic and viscosity measurement, and also for docking analysis. 4-MC exhibited good cytotoxicity on K562 cell line and the IC₅₀ value was calculated to be 30 µM. Furthermore, the mechanisms of apoptosis induction were determined morphologically by fluorescence dual staining with acridine orange and ethidium bromide and cell cycle analysis was based on DNA content, as well as the presence of phosphatidyl serine on the outside of the cells by the flow cytometric method. The results showed that 4-MC had potent cytotoxic activity via sub-G1 cell cycle arrest and induction of apoptosis. The experimental and simulation studies reported that 4-MC binds to ctDNA through groove binding mode with the binding constant (K_b) of 2.5 × 10³ M⁻¹. These data represent a considerable anticancer potential of 4-MC and could be suggested for further pharmacological studies.

摘要

Chromene 及其衍生物通常在自然界中传播。基于杂环的化合物如色烯已显示出药理活性。由于抗癌活性等一些生物学特性，Chromene 衍生物至关重要。CML，慢性粒细胞白血病，是一种致命的恶性肿瘤，由细胞凋亡抗性决定，包含费城染色体。诱导细胞凋亡是癌症治疗的主要方法之一。本研究中，苯并色烯衍生物，2-氨基-4-(4-甲氧基苯基)-4 H在人类白血病 K562 细胞系中测试了 -benzochromene-3-carbonitrile (4-MC) 的细胞毒性和凋亡诱导活性。MTT生长抑制试验用于确定细胞生长和存活。此外，通过一些光谱和粘度测量以及对接分析来评估 4-MC 与双螺旋 DNA 的结合属性。4-MC 对 K562 细胞系和 IC ₅₀表现出良好的细胞毒性计算值为 30 µM。此外，通过用吖啶橙和溴化乙锭荧光双重染色从形态学上确定细胞凋亡诱导的机制，细胞周期分析基于 DNA 含量，以及通过流式细胞术方法在细胞外部存在磷脂酰丝氨酸。结果表明，4-MC通过亚 G1 期细胞周期阻滞和诱导细胞凋亡具有有效的细胞毒活性。实验和模拟研究表明，4-MC 通过凹槽结合模式与 ctDNA 结合，结合常数 ( K _b ) 为 2.5 × 10 ³ M ^-1. 这些数据代表了 4-MC 的相当大的抗癌潜力，可以建议用于进一步的药理学研究。