多花蒿中流份 B 和 E 的抗肝癌活性成分研究

	多花蒿中流份 B 和 E 的抗肝癌活性成分研究
	张鑫
导师	陈纪军
关键词	多花蒿，多花蒿素 A~L，吉玛烷型倍半萜，抗肝癌细胞毒活性 Artemisia myriantha, artemyrianosins A~L, germacrane sesquiterpenoids, antihepatoma activity
摘要	肝细胞癌 (HCC) 是肝癌中最普遍、最严重的一种类型，其发病隐匿、进展快、预后差，而成为死亡率全球第三的癌症。HCC给全人类造成了极大的疾病负担，其防治是急需解决的问题。目前，临床上用于治疗 HCC 的药物主要有 7 个，其中索拉非尼 (sorafenib)、瑞格非尼 (regorafenib)、乐伐替尼 (Lenvatinib)、卡博替尼 (cabozantinib) 等 4 个化学合成抗肝癌药物主要作用于受体酪氨酸激酶，纳武单抗和帕姆单抗属于 PD-1 抑制剂，雷莫芦单抗为抗血管生成抑制剂，它们虽然取得了显著的临床疗效，但易产生耐药性和毒副反应，不能满足临床需求，因此急需开发用于治疗 HCC 的新药。多花蒿 (Artemisia myriantha) 系蒿属菊科植物，具有清热、祛暑、凉血、止血等功效，在传统中药中常用于治疗月经过多和炎症性疾病。本研究组前期对多花蒿的活性部位 C 和 D 中分离得到了系列化合物，大多数化合物对三株肝癌细胞 HepG2、SMMC-7721 和 Huh7 具有细胞毒活性。为了继续研究多花蒿中抗肝癌细胞毒活性的化合物，利用硅胶、MCI gel CHP 20P、Sephadex LH-20 等柱色谱以及制备型和半制备型 HPLC，对活性流份 Fr. B 和 Fr. E 进行分离纯化得到 39 个化合物，根据高分辨质谱 (HRESIMS)、红外光谱 (IR)、核磁共振谱 (1D和2D NMR)、旋光值、实验值和计算值 ECD 谱、X-ray 单晶衍射等现代波谱技术鉴定了他们的结构，其中新化合物 12 个，化合物结构类型包括 15 个吉玛烷型 (1~15)、4 个愈创木烷型 (16~19)、4 个桉烷型 (20~23)；木脂素类化合物 11 个 (24~34)，其他类型化合物 5 个(35~39)。12 个新化合物命名为多花蒿素 A~L (artemyrianosins A~L, 1~10, 16~17)，其中 4 个化合物的绝对构型是通过 X-ray 单晶衍射所确定。在浓度为200 和100 μM 时，对所得倍半萜类化合物进行了抗肝癌细胞毒活性筛选。结果表明， 5 个化合物对三株肝癌细胞 HepG2、Huh7 和 SK-Hep-1 具有抑制作用，其抑制率均大于 50%。随后对活性化合物进行剂量-效应测定得到化合物 1~3、7 和 10 对三株肝癌细胞的 IC50 值在 43.7 至 89.3 μM 之间。其中，多花蒿素 C (3) 的活性最好，其 IC50 值分别为 43.7 μM (HepG2)、47.9 μM (Huh7)、44.9 μM (SK-Hep-1)。以上研究结果进一步说明了多花蒿中抗肝癌活性成分主要为吉玛烷型倍半萜，为从多花蒿里寻找抗肝癌活性药物小分子提供了初步化学和药理学依据。; Hepatocellular carcinoma (HCC) is one of the most serious and common type of liver cancer. Its clinical features are insidious onset, rapid progress, and poor prognosis, which results in the third highest mortality rate among all cancers in the world. HCC caused great disease burden around the world, the prevention and treatment of which have drawn much attention from all sectors of society. Clinically, four synthetic drugs (sorafenib, regorafenib, lenvatinib and cabozantinib) and three monoclonal antibody medicants (nivolumab, pembrolizumab and ramucirumab) are used to treat HCC. Although these drugs have obtained significant achievements, there are still some inevitable drawbacks, such as drug resistance and serious toxicity. Therefore, new drugs are urgently needed for the treatment of HCC. Artemisia myriantha, belonging to the family Asteraceae, is commonly used for treating menorrhagia and inflammatory diseases in traditional Chinese medicine. Our previous investigation reported a series of compounds from the active fractions C and D of A. myriantha, and most of them showed cytotoxic activity against HepG2, SMMC-7721 and Huh7. During our continuous search for antihepatic constituents from A. myriantha, 39 compounds were isolated and elucidated from the active fractions B and E by various chromatographic methods including silica gel, MCI gel CHP 20P, Sephadex LH-20, preparative and semi-preparative HPLC. Based on modern spectroscopic techniques such as HRESIMS, IR, 1D and 2D NMR experimental and calculated ECD spectra, and X-ray single crystal diffraction, all compounds were classified as 15 germacranes (1~15), four guaianes (16~19), four eudesmanes (20~23), 11 lignans (24~34), and five other types (35~39). Among them, 12 new compounds were named as artemyrianosins A~L (1~10 and 16~17), and the absolute configurations of four compounds were determined by X-ray single-crystal diffraction. All sesquiterpenoids were tested for their cytotoxic activity against three human hepatoma cell lines (HepG2, Huh7 and SK-Hep-1) at the concentrations of 200 and 100 μM, in which five compounds displayed inhibitory effects with inhibitory ratios higher than 50%. The dose-response curves of the active compounds were further investigated to yield their respective IC50 values, which suggested compounds 1?3, 7, and 10 manifested cytotoxicity with IC50 values ranging from 43.7 to 89.3 μM. Among them, the most active compound 3 exhibited activity against three human hepatoma cell lines with IC50 values of 43.7 μM (HepG2), 47.9 μM (Huh7), and 44.9 μM (SK-Hep-1). The above results further demonstrated that the antihepatoma components in A. myriantha were mainly germacrene-type sesquiterpenoids, which provided preliminary chemical and pharmacological backgrounds for the search of antihepatoma candidates from A. myriantha.
语种	中文
	2022-05
学位授予单位	中国科学院大学
文献类型	学位论文
条目标识符	http://ir.kib.ac.cn/handle/151853/75185
专题	昆明植物所硕博研究生毕业学位论文
推荐引用方式 GB/T 7714	张鑫. 多花蒿中流份 B 和 E 的抗肝癌活性成分研究[D]. 中国科学院大学,2022.

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