新型LSD1抑制剂抗肿瘤作用及分子机制研究

The LSD1 is the first to be discovered demethylase, which plays an important role in tumorigenesis and development, and it has become one of the most potential targets in anticancer drug research and development. In our preliminary study, we targeted LSD1 and obtained a novel small molecule compounds ZY0511 by computer-aided drug design, synthesis and activity screening. It can significantly inhibit LSD1 activity (IC50 1.4nM), increase H3K4 and H3K9 methylation levels and obviously inhibit the cancer cells proliferation both in vitro and in vivo. However, its anticancer mechanism still remains unclear and needs further investigation. Base on the results of preliminary studies, we will continue studying the specific and feature of what ZY0511 Effects on LSD1; whether the ZY0511 will active or inhibit the downstream target genes of the LSD1; the mechanism and effect of ZY0511 on cancer stem cell; how the ZY0511 exerts its antitumor effect by acting on the pathway related to the LSD1. By means of researching the anticancer mechanism of ZY0511 thoroughly ,we can provide more theoretical basis for its further applications and the function of LSD1 as a cancer diagnosis and therapeutic targets. Moreover, ZY0511 has also become a molecular probe tool to help reveal the new mechanism of LSD1 on tumor regulation, and provide a powerful theoretical basis for LSD1 as tumor diagnosis and therapy.

组蛋白赖氨酸特异性去甲基化酶 1 （LSD1）是第一个被发现的去甲基化酶，其在肿瘤发生发展中起重要作用，已成为具重要发展前景的抗肿瘤新靶点。我们前期研究以LSD1为靶标，通过计算机辅助药物设计，自行合成及筛选得到了结构全新的小分子化合物ZY0511。其可显著抑制LSD1活性（IC50 1.4nM），升高H3K4和H3K9甲基化水平，体内外显著抑制肿瘤增殖，但尚需对其抗肿瘤机制进行深入研究。本项目将在前期研究基础上，结合结构生物学，代谢组学等技术研究ZY0511对LSD1作用特异性和作用特点；ZY0511对LSD1下游直接靶基因激活或抑制作用；ZY0511对肿瘤干细胞的增殖抑制作用和机制；ZY0511如何通过作用于与LSD1有交叉对话的相关通路发挥抗肿瘤作用。上述研究揭示ZY0511抗肿瘤作用机制，为其应用提供理论基础；也揭示LSD1调控肿瘤的新机制，为LSD1作为肿瘤诊断和治疗靶点提供依据

组蛋白赖氨酸特异性去甲基化酶 1 （Lysine specific demethylase 1, LSD1）是首个被发现的组蛋白去甲基化酶，其在肿瘤发生发展中起重要作用，已成为具重要发展前景的抗肿瘤新靶点。本课题组前期以LSD1为靶标，通过计算机辅助药物设计、自行合成及筛选得到了结构全新的小分子化合物ZY0511。其可显著抑制LSD1活性和体外肿瘤细胞增殖。本项目在前期研究基础上，结合转录组学等技术研究了ZY0511对LSD1作用特点、深入研究ZY0511体内外抗肿瘤作用和机制。研究结果显示，ZY0511可特异性抑制LSD1活性并与LSD1发生直接相互作用。ZY0511体外显著抑制多种不同组织来源人恶性肿瘤细胞株包括人结直肠癌、宫颈癌和肝癌等增殖，而且此抑制作用是LSD1依赖的。体内研究进一步确认ZY0511（15-60 mg/kg，每日一次，腹腔或者口服给药）可以显著抑制裸鼠皮下结直肠癌、宫颈癌、肝癌等移植瘤模型肿瘤生长，抑瘤率最高达63%。此外，LSD1过表达还与结直肠患者对5-FU耐药相关，ZY0511在体内外可以增强5-FU抗结直肠作用。ZY0511在体外可增强紫杉醇对人宫颈癌细胞增殖抑制作用。作用机制研究显示ZY0511增加H3K4和H3K9甲基化，从而调控肿瘤细胞增殖、凋亡和周期等相关基因的表达。进一步深入研究发现了新的LSD1下游直接调控靶基因。在宫颈癌中，Ddit4是LSD1直接作用下游靶基因；在肝癌中，GADD45B是LSD1直接作用下游靶基因。ZY0511通过调控上述基因表达从而发挥抗肿瘤作用。综上，本研究揭示了ZY0511具有显著抗肿瘤作用，明确了ZY0511抗肿瘤作用机制，为其应用提供理论基础。同时，研究帮助揭示LSD1调控肿瘤的新机制，为LSD1作为肿瘤诊断和治疗靶点提供更有力理论依据。