Department of Chemistry, University of Alberta, Edmonton, AB T6G 2G2, Canada; Department of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, Alexandria University, Alexandria, Egypt.
Department of Chemistry, University of Alberta, Edmonton, AB T6G 2G2, Canada.
J Pharm Biomed Anal. 2018 Aug 5;157:226-234. doi: 10.1016/j.jpba.2018.05.019. Epub 2018 May 17.
Hepatocellular carcinoma is one of the most common malignant tumors in the world. Chronic hepatitis B and C infections are the most common etiologies of hepatocellular carcinoma worldwide. In this study, we explore the potential DNA damaging effect of some FDA-approved antiviral drugs which may be able to serve as anticancer agents for hepatocellular carcinoma, in order to better elucidate their mode of action. Five antiviral drugs were selected; ribavirin, sofosbuvir, tenofovir disoproxil fumerate, daclatasvir and ledipasvir. Several methods, including absorption spectroscopy, MALDI-TOF mass spectrometry and fluorimetric analysis using the EvaGreen (EG) intercalating dye, were used to probe the drug-induced DNA damage. Results show that only daclatasvir and ledipasvir induced DNA damage. Absorption spectroscopy showed hyperchromicity in the 260-nm DNA absorption band of DNA samples incubated with each drug, indicating disruption of the double-strand structure. Mass spectra for DNA samples incubated with each of the two drugs showed a disappearance of the DNA molecular ion peak with a concomitant appearance of peaks with smaller m/z, indicating DNA strand breaks. EG fluorescence was observed to decrease with increasing incubation time of daclatasvir and ledipasvir with DNA, indicating that the EG detaches from the DNA, likely due to DNA damage. All of these results are consistent with DNA damage, proposed as oxidative damage to both nucleobase and deoxyribose moieties of DNA as the mode of action for these two drugs. Moreover, these results are dependent on the antiviral drug concentration and show that DNA regions rich in guanine are affected more than other regions by these two drugs. Therefore, such antiviral drugs may present a promising therapeutic alternative to the currently used anticancer agents, especially for hepatitis B and C patients with hepatocellular carcinoma resistant to conventional treatment approaches.
肝细胞癌是世界上最常见的恶性肿瘤之一。慢性乙型和丙型肝炎感染是全球肝细胞癌最常见的病因。在这项研究中，我们探索了一些已获 FDA 批准的抗病毒药物对 DNA 可能具有的潜在损伤作用，这些药物可能成为治疗肝细胞癌的抗癌药物，以便更好地阐明它们的作用机制。选择了五种抗病毒药物：利巴韦林、索非布韦、富马酸替诺福韦二吡呋酯、达卡他韦和来迪派韦。使用吸收光谱法、基质辅助激光解吸电离飞行时间质谱法（MALDI-TOF MS）和使用 EvaGreen（EG）嵌入染料的荧光分析等几种方法来探测药物诱导的 DNA 损伤。结果表明，只有达卡他韦和来迪派韦诱导了 DNA 损伤。吸收光谱法显示，与每种药物孵育的 DNA 样品在 260nm 的 DNA 吸收带处出现增色现象，表明双链结构被破坏。与两种药物孵育的 DNA 样品的质谱显示，DNA 分子离子峰消失，同时出现较小 m/z 的峰，表明 DNA 链断裂。随着达卡他韦和来迪派韦与 DNA 孵育时间的增加，EG 荧光观察到减少，表明 EG 从 DNA 上脱离，可能是由于 DNA 损伤。所有这些结果都与 DNA 损伤一致，这两种药物的作用机制被提议为对 DNA 的碱基和脱氧核糖部分的氧化损伤。此外，这些结果取决于抗病毒药物浓度，表明这些药物对富含鸟嘌呤的 DNA 区域的影响比对其他区域的影响更大。因此，这些抗病毒药物可能为目前使用的抗癌药物提供一种有前途的治疗选择，特别是对于乙型和丙型肝炎病毒感染且对传统治疗方法耐药的肝细胞癌患者。
