PDGFR小分子抑制剂GZD856体内外抗肺癌作用及其机制研究

发布时间：2018-05-20 00:28

本文选题：血小板衍生生长因子受体抑制剂 + GZD856　；参考：《暨南大学》2016年博士论文

【摘要】：血小板衍生生长因子受体(PDGFRs)通过影响肿瘤自分泌生长因子和对癌相关成纤维细胞的招募,在人类肿瘤的恶性增殖中发挥重要作用。本论文开展PDGFR小分子抑制剂GZD856体内外抗肺癌作用及其机制研究。本论文首先利用ELISA方法研究GZD856体外激酶抑制活性,MTT法研究体外细胞增殖抑制活性,流式细胞术研究对细胞周期和凋亡的影响,然后使用Western Blot方法研究其体外作用机制,进而使用移植瘤模型及原位模型研究体内抗肿瘤增殖及抗转移作用,Western Blot及免疫组化方法研究其体内作用机制。研究结果表明:GZD856能够抑制PDGFRα/β的激酶活性,并可以体内外阻断PDGFRα/β及其下游信号分子的活化;细胞活性结果显示其对H1703PDGFRα+/PDGFRβ-细胞表现出较强的抗增殖活性,而对A549PDGFRα-/PDGFRβ+细胞的活性较差;体内结果显示,GZD856在30 mg/kg剂量基本可以完全抑制H1703的增殖,同时对A549细胞也显示出较好的抗增殖活性,进一步的结果证实其也能抑制A549细胞肺部克隆的体积和肝脑转移;深入的机制研究证实GZD856对A549的体内治疗作用与其抑制PDGFR介导的肿瘤微环境中癌相关成纤维细胞的招募及相关生长因子的分泌有关。同时GZD856在大鼠体内表现出优秀的口服生物利用度(78%)、半衰期(22 hr)及血药浓度(Cmax,899.5μg/L),在ICR小鼠体内具有合适的LD50(po,497.0 mg/kg)。综上所述,本论文通过不同生物学手段鉴定了一个结构全新的PDGFRα/β小分子抑制剂。GZD856是一个优秀的治疗肺癌的候选先导化合物。
[Abstract]:Platelet-derived growth factor receptor PDGFRsplays an important role in the malignant proliferation of human tumors by affecting tumor autocrine growth factor and recruitment of cancer-related fibroblasts. In this paper, the anti-lung cancer effect and its mechanism of PDGFR small molecule inhibitor GZD856 in vitro and in vivo were studied. In this paper, ELISA was used to study the inhibitory activity of GZD856 on cell proliferation, flow cytometry was used to study the effect of flow cytometry on cell cycle and apoptosis, and then Western Blot was used to study its mechanism in vitro. Then the tumor transplantation model and in situ model were used to study the anti-tumor proliferation and anti-metastasis effects in vivo. Western Blot and immunohistochemical method were used to study the mechanism of its action in vivo. The results showed that: GZD856 could inhibit the kinase activity of PDGFR 伪 / 尾, and block the activation of PDGFR 伪 / 尾 and its downstream signaling molecules in vitro and in vivo, and the cell activity showed strong antiproliferative activity on H1703PDGFR 伪 / PDGFR 尾 cells, but poor on A549PDGFR 伪 / PDGFR 尾 cells. The results in vivo showed that GZD856 could completely inhibit the proliferation of H1703 at the dose of 30 mg/kg, and also showed good antiproliferative activity on A549 cells. The further results confirmed that GZD856 could also inhibit the volume of lung clone and hepatic and brain metastasis of A549 cells. Further studies have confirmed that the in vivo therapeutic effect of GZD856 on A549 is related to its inhibition of PDGFR mediated recruitment of cancer-related fibroblasts and secretion of related growth factors in tumor microenvironment. At the same time, GZD856 showed excellent oral bioavailability in rats with a half-life of 22 h) and plasma concentration of Cmax.899.5 渭 g / L, and a suitable LD50 pobutamine 497.0 mg / kg 路kg ~ (-1) in ICR mice. In conclusion, a novel PDGFR 伪 / 尾 small molecule inhibitor. GZD856 was identified as an excellent candidate leading compound for the treatment of lung cancer by different biological methods.
【学位授予单位】：暨南大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R734.2

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