萘普替尼靶向表皮生长因子受体不同突变亚型的抗肿瘤作用研究

目的 研究萘普替尼对表皮生长因子受体（EGFR）不同突变亚型荷瘤小鼠肿瘤生长的影响。方法 建立野生型EGFR人表皮鳞癌A431、L858R/T790M双突变EGFR非小细胞肺癌H1975、Del19突变型EGFR非小细胞肺癌HCC827及HER2高表达的胃癌N87的裸鼠移植瘤模型，待肿瘤长至100 mm³左右将小鼠分组，A431荷瘤小鼠分组：对照组、阿法替尼12 mg/kg和萘普替尼3.50、1.75、0.87、0.29 mg/kg；H1975荷瘤小鼠分组：对照组、阿法替尼30 mg/kg和萘普替尼7.00、3.50、1.75 mg/kg；HCC827荷瘤小鼠分组：对照组、阿法替尼12 mg/kg和萘普替尼3.50、1.75、0.87、0.60、0.29 mg/kg；N87荷瘤小鼠分组：对照组、阿法替尼12 mg/kg和萘普替尼7.00、3.50、1.75、0.87、0.29 mg/kg，每组10只动物。分组后即开始ig给药，对照组给予同体积去离子水，A431、H1975、HCC827、N87荷瘤小鼠分别给药14、21、7、14 d。观察相对肿瘤体积（RTV）、肿瘤增殖率及对肿瘤质量的抑制率，在体考察萘普替尼的抗肿瘤作用。结果 A431荷瘤小鼠：与对照组比较，萘普替尼各剂量组RTV均显著减小（P<0.01），肿瘤增殖率分别为26.6%、32.5%、34.8%、42.2%；瘤质量均显著降低（P<0.01），抑瘤率范围在48.3%~73.9%；0.87 mg/kg为起效剂量，3.5 mg/kg剂量与阿法替尼12 mg/kg效果相当，3.5mg/kg组小鼠体质量显著降低（P<0.01）。H1975荷瘤小鼠：与对照组比较，萘普替尼各剂量组RTV均显著降低（P<0.05、0.01），肿瘤增殖率分别为14.5%、38.2%、65.3%；3.5 mg/kg为起效剂量，与阿法替尼30 mg/kg作用相当；7.0、3.5 mg/kg组小鼠体质量显著降低（P<0.05、0.01）。HCC827荷瘤小鼠：与对照组比较，萘普替尼3.50、1.75、0.87、0.60 mg/kg组RTV降低极为显著（P<0.01），肿瘤增殖率均小于40%；瘤质量呈不同程度地降低，抑瘤率范围52.9%~89.7%；0.6 mg/kg为起效剂量，1.75 mg/kg剂量与阿法替尼12 mg/kg作用相当，3.5、1.75 mg/kg组小鼠体质量显著降低（P<0.01）。N87荷瘤小鼠：萘普替尼7.00、3.50、1.75 mg/kg组RTV及瘤质量均显著低于对照组（P<0.05、0.01），且肿瘤增殖率均小于40%，抑瘤率范围54.9%~95.00%；1.75 mg/kg为起效剂量，3.5 mg/kg与阿法替尼12 mg/kg作用相当；7.0、3.5 mg/kg组小鼠体质量显著降低（P<0.01）。结论 萘普替尼具有明显的抗肿瘤作用，其中对EGFR野生型或单突变型荷瘤抗肿瘤作用较好。关键词：萘普替尼；表皮生长因子受体；表皮生长因子受体酪氨酸激酶抑制剂；突变；非小细胞肺癌

Anti-tumor effect of neptinib targeting different mutant subtypes of EGFR

Objective To study the effect of neptinib on the growth of tumors in mice bearing different EGFR mutant subtypes. Methods Xenograft models of wild-type EGFR human epidermal squamous cell carcinoma cell line A431, double mutant EGFR (L858R/T790M) non-small cell lung cancer cell line H1975, Del19 mutant EGFR non-small cell lung cancer cell line HCC827 and HER2 highly expressed gastric cancer cell line N87 in nude mice were established. The mice were divided into groups when the tumor grew to 100 mm³. A431 mice were divided into control group, alfatinib 12 mg/kg, naphtinib 3.50, 1.75, 0.87, 0.29 mg/kg; H1975 mice were divided into control group, alfatinib 30 mg/kg, naphtinib 7.00, 3.50, 1.75 mg/kg; HCC827 mice were divided into control group, alfatinib 12 mg/kg, naphtinib 3.50, 1.75, 0.87, 0.60, 0.29 mg/kg; N87 mice were divided into control group, alfatinib 12 mg/kg, naphtinib 7.00, 3.50, 1.75, 0.87, 0.29 mg/kg. Ten animals in each group. The mice in control group were given deionized water of the same volume. The mice bearing A431, H1975, HCC827 and N87 were ig given for 14, 21, 7 and 14 days respectively. To observe the relative tumor volume (RTV), the tumor growth rate and the tumor inhibition rate, which aims to investigate the antitumor effect of neptinib in vivo. Results A431 tumor-bearing mice:Compared with the control group, the RTV of each dose of Naprotinib decreased significantly (P<0.01), the proliferation rate of tumors was 26.6%, 32.5%, 34.8%, 42.2%, the tumour quality was significantly reduced (P<0.01), the inhibition rate ranged from 48.3% to 73.9%; 0.87 mg/kg was the effective dose, 3.5 mg/kg was equivalent to 12 mg/kg of Alphatinib, and the body weight of mice in 3.5 mg/kg group was significantly decreased (P<0.01). H1975 tumor-bearing mice:Compared with the control group, the RTV of each dose group of Naprotinib decreased significantly (P<0.05, 0.01), the proliferation rate of tumor was 14.5%, 38.2%, 65.3%, 3.5 mg/kg was the effective dose, which was equivalent to 30 mg/kg of Alfatinib, and the body weight of mice in 7.0 and 3.5 mg/kg groups decreased significantly (P<0.05, 0.01). HCC827 tumorbearing mice:Compared with the control group, the RTV of Naprotinib 3.50, 1.75, 0.87, 0.60 mg/kg group decreased significantly (P<0.01), and the proliferation rate of tumors was less than 40%. The tumour quality decreased in varying degrees, and the inhibition rate ranged from 52.9% to 89.7%. The effective dose was 0.6 mg/kg, and the dose of 1.75 mg/kg was equivalent to that of alfatinib 12 mg/kg, and the body weight of mice in 3.5 and 1.75 mg/kg groups decreased significantly (P<0.01). N87 tumor-bearing mice:RTV and tumor mass of naphtinib 7.00, 3.50, 1.75 mg/kg group were significantly lower than those of control group (P<0.05, 0.01), and the tumor proliferation rate was less than 40%, the inhibition rate ranged from 54.9% to 95.00%. 1.75 mg/kg was the effective dose, and the effect of 3.5 mg/kg was the same as that of afatinib 12 mg/kg; the body mass of mice in 7.0 and 3.5 mg/kg groups decreased significantly (P<0.01). Conclusion Neptinib has obvious anti-tumor effect, especially on tumors with wild-type or single-mutant EGFR.