Pylina Yana I, Shadrin Dmitry M, Shevchenko Oksana G, Startseva Olga M, Velegzhaninov Igor O, Belykh Dmitry V, Velegzhaninov Ilya O
Institute of Biology of Komi Science Centre of Ural Branch of RAS, Syktyvkar 167982, Russia.
Institute of Chemistry of Komi Science Centre of Ural Branch of RAS, Syktyvkar 167982, Russia.
Int J Mol Sci. 2017 Jan 5;18(1):103. doi: 10.3390/ijms18010103.
In the present work, we investigated the dark and photoinduced cytotoxic activity of the new chlorophyll-a derivatives which contain the substituents of oligoethylene glycol on the periphery of their macrocycles. These compounds were tested using human cell lines to estimate their potential as photosensitizers for photodynamic therapy of cancer. It was shown that all the tested compounds have expressed photoinduced cytotoxic activity in vitro. Detailed study of the biological activity of one of the most perspective compound in this series-pyropheophorbide-a 17-diethylene glycol ester (Compound ) was performed. This new compound is characterized by lower dark cytotoxicity and higher photoinduced cytotoxicity than previously described in a similar compound (DH-I-180-3) and clinically used Photolon. Using fluorescent microscopy, it was shown that Compound quickly penetrates the cells. Analysis of caspase-3 activity indicated an apoptosis induction 40 min after exposure to red light (λ = 660 nm). The induction of DNA damages and apoptosis was shown using Comet assay. The results of expression analysis of the stress-response genes indicate an activation of the genes which control the cell cycle and detoxification of the free radicals after an exposure of HeLa cells to Compound and to red light. High photodynamic activity of this compound and the ability to oxidize biomolecules was demonstrated on nuclear-free mice erythrocytes. In addition, it was shown that Compound is effectively activated with low energy 700 nm light, which can penetrate deep into the tissue. Thus, Compound is a prospective substance for development of the new drugs for photodynamic therapy of cancer.
在本研究中，我们研究了新型叶绿素-a衍生物的暗细胞毒性和光诱导细胞毒性活性，这些衍生物在其大环外围含有低聚乙二醇取代基。使用人类细胞系对这些化合物进行了测试，以评估它们作为癌症光动力疗法光敏剂的潜力。结果表明，所有测试化合物在体外均表现出光诱导细胞毒性活性。对该系列中最具前景的化合物之一——焦脱镁叶绿酸-a 17-二甘醇酯（化合物 ）的生物活性进行了详细研究。这种新化合物的特点是，与先前描述的类似化合物（DH-I-180-3）和临床使用的Photolon相比，其暗细胞毒性较低，光诱导细胞毒性较高。使用荧光显微镜观察发现，化合物 能迅速穿透细胞。对半胱天冬酶-3活性的分析表明，在暴露于红光（λ = 660 nm）40分钟后诱导了细胞凋亡。使用彗星试验显示了DNA损伤和细胞凋亡的诱导。应激反应基因表达分析的结果表明，在HeLa细胞暴露于化合物 和红光后，控制细胞周期和自由基解毒的基因被激活。在无核小鼠红细胞上证明了该化合物的高光动力活性和氧化生物分子的能力。此外，还表明化合物 能被低能量的700 nm光有效激活，这种光可以深入组织。因此，化合物 是开发用于癌症光动力治疗新药的一种有前景的物质。
