Evodiamine-inspired dual inhibitors of histone deacetylase 1 (HDAC1) and topoisomerase 2 (TOP2) with potent antitumor activity

A great challenge in multi-targeting drug discovery is to identify drug-like lead compounds with therapeutic advantages over single target inhibitors and drug combinations. Inspired by our previous efforts in designing antitumor evodiamine derivatives, herein selective histone deacetylase 1 (HDAC1) and topoisomerase 2 (TOP2) dual inhibitors were successfully identified, which showed potent in vitro and in vivo antitumor potency. Particularly, compound 30a was orally active and possessed excellent in vivo antitumor activity in the HCT116 xenograft model (TGI = 75.2%, 150 mg/kg, p.o.) without significant toxicity, which was more potent than HDAC inhibitor vorinostat, TOP inhibitor evodiamine and their combination. Taken together, this study highlights the therapeutic advantages of evodiamine-based HDAC1/TOP2 dual inhibitors and provides valuable leads for the development of novel multi-targeting antitumor agents.     

吴茱萸碱羟丙基-β-环糊精包合物大鼠在体肠吸收特征

[摘要] 目的 研究吴茱萸碱羟丙基-β-环糊精包合物(EHD)大鼠在体肠吸收特征。方法 制备吴茱萸碱羟丙基-β-环糊精包合物,并测定其理化性质;将健康SD雄性大鼠,随机分为2组,运用单向肠灌流模型对大鼠各肠段吴茱萸碱(EVO)的吸收情况进行考察,使用HPLC法（ C18柱(250 mm×4.6 mm,5 μm),流动相为甲醇:水=75:25(V/V),流速为1.0 mL/min,检测波长为225 nm,柱温35 ℃）测定EVO的含量,并对吸收速率常数(Ka)及有效渗透系数(Peff)进行计算。结果 EHD的傅里叶-红外扫描图谱中,吴茱萸碱特征吸收峰减弱;EHD的差示扫描量热图谱中,吴茱萸碱的吸热峰明显减小;EHD的电镜下形态与物理混合物明显的不同;肠循环液中吴茱萸碱的回收率与精密度符合要求;EHD在十二指肠、空肠、回肠、结肠中的Ka值分别为EVO的9.07、16.22、11.04、28.86倍,差异具统计学意义(P＜0.05)。EHD在十二指肠、空肠、回肠、结肠中的Peff值分别为EVO的2.41、1.52、1.82、1.09倍,在十二指肠处差异有统计学意义（P＜0.05）,其他肠段处差异均无统计学意义（P＞0.05）。结论 EHD能使EVO在大鼠体内的肠吸收得到明显的改善。     

Discovery of Novel Multiacting Topoisomerase I/II and Histone Deacetylase Inhibitors

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Atypical apoptosis in L929 cells induced by evodiamine isolated from Evodia rutaecarpa

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HPLC法同时测定吴茱萸中吴茱萸碱、吴茱萸次碱与吴茱萸内酯的含量

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Abstract：

Objective To investigate the effects of evodiamine on autophagy of human colon a cleno carcinoma lovo cells, and to explore the role and mechanism of autophagy which was induced by evodiamine (EVO). Methods MTT assay combined with the morphologic changes were used to observe the cell viability. Monodansylcadaverine was used to detect autophagy by fluorospectrophotometer and the confocal laser fluorescence microscopy respectively. Immunoblotting assay was used to observe the microtubule-associated protein 1 light chain 3. Finally, evodiamine combined with 3-methyladenine to detect the cell viability with MTT assay and the apoptosis with the flow cytometry, respectively.Results Evodiamine inhibited the viability of Lovo cells in dose-dependent manner ( P ＜ 0. 05 ), especially in 60 μmol/L that was obviously(60% ). Further more, the cell lysis and cell gap widened was observed by the light microscope. Evo triggered the autophagy, and after inhibition the autophagy by 3-MA, the killing capacities of the Evo was enhanced ( P ＜ 0. 01 ). However, autophagy prohibited the apoptosis pathways.Conclusions Evodiamine can trigger the autophagy, which might play a self-defense role in evodiamineinduced cell death. The cytototoxicity of evodiamine can be augmented by the autophagy inhibitors. The joint application of autophagy regulators with the chemotherapeutic agents might enhance the cell killing effects of chemotherapeutic drugs and show a potent role in cancer drug resistance.     

Atypical apoptosis in L929 cells induced by evodiamine isolated from Evodia rutaecarpa

Two alkaloids, evodiamine and rutaecarpine, isolated from the dried fruits of Evodia rutaecarpa Bentham were evaluated in vitro for antiproliferation activity on tumor cells versus human peripheral blood mononuclear cells (PBMC). Evodiamine had more potent cytotoxic effects on five tumor cell lines (human malignant melanoma A375-S2, human cervical cancer HeLa, human breast adenocarcinoma MCF7, human acute monocytic leukemia THP-1, murine fibrosarcoma L929) than rutaecarpine. Moreover, evodiamine did not affect PBMC viability for a 36?h culture period. Although apoptotic bodies were observed in evodiamine-treated L929 cells stained with Hoechst 33258, DNA fragmentation as a hallmark of apoptosis was not found. Caspases were involved in the protection of L929 cells against cell death. Evodiamine initiated atypical apoptosis in L929 cells by cycle arrest at the G0/G1 phase.