MHY1485 is an mTOR activator that is cell-permeable. MHY1485 inhibits autophagosome and lysosome fusion, leading to accumulation of LC3II proteins and increased autophagosomes, thereby inhibiting cellular autophagy.

mTOR:~2 μM

方法：正常大鼠肝细胞 Ac2F 用 MHY1485 (0.5-2 µM) 处理 1-12 h，通过 Western Blot 检测靶点蛋白表达水平。
结果：MHY1485 显著增加了 LC3II/LC3I 比值，呈剂量依赖性和时间依赖性。[1]
方法：肿瘤细胞系 CT26 和 LLC 用 MHY1485 (1-10 µM) 和 X-irradiation (0-6 Gy) 处理 5 天，检测细胞数目。
结果：在 CT26 中，与未处理相比，单独用 MHY1485 处理的细胞生长明显延迟；与单独辐射相比，MHY1485 和辐射的联合治疗显著延迟了细胞生长。在LLC中，MHY1485 处理在非照射和照射条件下都显著延迟了细胞生长。结果表明，MHY1485 在体外抑制肿瘤细胞生长，无论是单独给药还是与辐射联合给药。[2]

方法：为研究自噬调节对视网膜神经节细胞的影响，将 MHY1485 (10 mg/kg) 腹腔注射给糖尿病小鼠，每天一次，持续 10 天。
结果：MHY1485 治疗成功激活了 3 m STZ 小鼠神经视网膜中的 mTOR，MHY1485 治疗导致自噬受到抑制。与正常小鼠视网膜一样，MHY1485 处理的小鼠视网膜仅在 NFL 和 GCL 中显示 GFAP 信号。[3]

Ovaries from mice at day10 of age are treated with 10 μM MHY1485 for 3h and proteins are extracted using M-PER Mammalian Protein Extraction Reagent containing a protease inhibitor cocktail. Protein concentrations in supernatants are determined by the bicinchoninic acid method. Equal amounts of protein lysates are loaded on 4-12% NuPAGE Bis-Tris gels in MOPS buffer and transferred to 0.45 μM pore nitrocellulose membranes[2].

MHY1485 is dissolved in DMSO and then diluted with appropriate media[3]. MC3T3-E1 cells are maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin and 100 mg/mL streptomycin at 37°C in a humidified atmosphere of 5% CO2. Having reached 70% confluence, the culture medium is switched to commercial osteogenic differentiation medium. MC3T3-E1 cells are cultured in the osteogenic differentiation medium for 14 days, following by culture in DMEM supplemented with varying concentrations of liraglutide for a further 14 days. MC3T3-E1 cells treated with 4 nM liraglutide are cultured in the presence or absence of Compound C or MHY1485. MC3T3-E1 cells maintained in DMEM for 28 days in the absence of any treatment are used as the negative control (NC); cells cultured in commercial osteogenic differentiation medium for 14 days and in DMEM without liraglutide for an additional 14 days are used as the positive control (PC)[3].