化合物C17通过抑制肿瘤干细胞抗胰腺癌的药效学及机制研究

The anti-tumor efficacy and mechanism of C17 in pancreatic cancer:inhibition of cancer stem-like cells

C17是一种包含2,4-二氨基喹唑啉和芳基哌嗪的小分子化合物。本研究首先利用分子对接(docking)考察C17与多巴胺D1受体(D1 dopamine receptor,D1DR)等5种DR亚型的结合情况,结果显示,在5个DR亚型中,C17最有可能与D1DR结合。随后采用胰腺癌细胞SW1990及PANC-1进行体内外实验:免疫荧光结果显示,C17可上调两种细胞中的D1DR表达;由细胞毒实验可知C17对SW1990、PANC-1细胞的IC50值分别为12.56和10.56μmol·L-1;克隆形成实验显示C17可以剂量依赖性地抑制两种细胞的克隆形成能力;分别采用CD133和ALDH标记SW1990和PANC-1细胞中肿瘤干细胞(cancer stem-like cell,CSC)亚群,并用流式细胞术证明C17可剂量依赖性地抑制两种细胞的CSC比例;将C17预处理的SW1990细胞接种至nu/nu裸鼠体内,结果显示C17可抑制其体内致瘤性,且与CSC比例相关;进一步体内实验显示,C17单用及与舒尼替尼(sunitinib,SUN)联用具有良好的体内药效和安全性。所有动物实验均严格遵循北京大学生物医学伦理委员会的规定。上述结果提示,C17可能具有用于胰腺癌治疗的潜力。

C17 is a small molecule containing 2,4-diaminoquinazoline and aryl piperazine.Docking was used to compare the affinity of C17 for five dopamine receptor(DR)subtypes.Pancreatic cancer SW1990 and PANC-1cell lines were used in in vitro and in vivo studies.The effect of C17 on the expression level of D1DR was investi‐gated by immunofluorescence.A cytotoxicity assay,clone formation assay and flow cytometry were used to investi‐gate the ability of C17 to inhibit on cell survival,clone formation,and to suppress cancer stem-like cells(CSCs)The ability to suppress tumorigenesis was investigated by inoculating nude mice with SW1990 cells pre-treated with different concentrations of C17.Finally,the anti-tumor efficacy and safety of C17 and its combination with the multitarget tyrosine kinase inhibitor sunitinib(SUN)were evaluated in SW1990 xenograft mice.Our results demonstrate that C17 is most likely to bind to D1DR among the five DR subtypes.D1DR expression was increased in C17-treated cells,which could be reversed by SCH23390,a D1DR-specific antagonist.The IC50values of C17on the survival of SW1990 and PANC-1 cells were 12.56 and 10.56μmol·L-1,respectively.C17 could suppress clone formation ability,CSC frequency and in vivo tumorigenesis in a dose-dependent manner.In the SW1990xenograft model,50 mg·kg-1of C17 could weakly inhibit the tumor growth,and the tumor volume with 50 mg·kg-1of C17 in combination with 10 mg·kg-1of SUN group was smaller than that in SUN 10 mg·kg-1,SUN 20 mg·kg-1group and gemcitabine(GEM)group.In addition,body weight,blood test,and organ index results showed good safety with all dosing regimens.All animal experiments were in strict accordance with the regulations of the Biomedical Ethics Committee of Peking University.C17 may be a promising candidate for the treatment of pancreatic cancer.