| Description | Leupeptin hemisulfate is a protease inhibitor with cell membrane-permeable, reversible, competitive, and oral activities. Leupeptin hemisulfate inhibits the activity of Cathepsin B, Cathepsin H, and Cathepsin L, and blocks fusion of amphipathic lysosomes. Leupeptin hemisulfate also has anti-inflammatory activity. |
| In vitro | 方法：SARS-CoV-2 感染的 Vero 细胞用 Leupeptin hemisulfate (0.06-200 µM) 处理 72 h，使用 RT-PCR 检测病毒 RNA 水平。结果：Leupeptin hemisulfate 可以抑制 Vero 细胞中 SARS-CoV-2 的 RNA 水平，EC50 值为 42.34 µM。[1]方法：SARS-CoV-2 假病毒感染的 Huh7 细胞用 Leupeptin hemisulfate (0.1-100 µM) 处理 24 h，使用 luciferase activity assay 检测假病毒感染情况。结果：Leupeptin hemisulfate 抑制 Vero 细胞中的假病毒感染，EC50 值为 39.29 µM。[2] |
| In vivo | 方法：为测定巨噬细胞自噬通量，将 Leupeptin hemisulfate (9-40 mg/kg in 0.5 mL PBS) 单次腹腔注射给 C57BL/6NCrl 小鼠。结果：Leupeptin hemisulfate 治疗后 LC3b 的积累率在肝脏最高，在脾脏最低。LC3a、ATG8/LC3b 同源物和 LC3b 相互作用蛋白 p62 以与 LC3b 相似的动力学降解。[3]方法：为检测寒冷是否激活自噬流量，将 Leupeptin hemisulfate (40 mg/kg) 单次腹腔注射给 C57B6 小鼠，1 h 后将小鼠在 4 ℃ 下冷暴露 1 h。结果：冷暴露小鼠在棕色脂肪中的 LC3-II 通量增加了 >2 倍。[4] |
| Synonyms | 亮肽素 |
| molecular weight | 475.59 |
| Molecular formula | C20H38N6O4·1/2H2SO4 |
| CAS | 103476-89-7 |
| Storage | keep away from moisture | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| Solubility | H2O: 87 mg/mL (182.9 mM) DMSO: 60 mg/mL (126.16 mM) Ethanol: 88 mg/mL (185 mM) |
| References | 1. Fu L, et al. Mechanism of Microbial Metabolite Leupeptin in the Treatment of COVID-19 by Traditional Chinese Medicine Herbs. mBio. 2021 Oct 26;12(5):e0222021. 2. Yang WL, et al. Potential drug discovery for COVID-19 treatment targeting Cathepsin L using a deep learning-based strategy. Comput Struct Biotechnol J. 2022;20:2442-2454. 3. Haspel J, et al. Characterization of macroautophagic flux in vivo using a leupeptin-based assay. Autophagy. 2011 Jun;7(6):629-42. 4. Martinez-Lopez N, et al. Autophagy in the CNS and Periphery Coordinate Lipophagy and Lipolysis in the Brown Adipose Tissue and Liver. Cell Metab. 2016 Jan 12;23(1):113-27. 5. Jiang T Y, Pan Y F, Wan Z H, et al. PTEN status determines chemosensitivity to proteasome inhibition in cholangiocarcinoma[J]. Science Translational Medicine. 2020, 12(562). |
| Citations | 1. Jiang T Y, Pan Y F, Wan Z H, et al. PTEN status determines chemosensitivity to proteasome inhibition in cholangiocarcinoma. Science Translational Medicine. 2020, 12(562). 2. Fu L, Shao S, Feng Y, et al. Mechanism of Microbial Metabolite Leupeptin in the Treatment of COVID-19 by Traditional Chinese Medicine Herbs. Mbio. 2021, 12(5): e02220-21. 3. Tao L, Cao Y, Wei Z, et al. Xanthatin triggers Chk1-mediated DNA damage response and destabilizes Cdc25C via lysosomal degradation in lung cancer cells. Toxicology and Applied Pharmacology. 2017 Dec 15;337:85-94 4. Tao L, Zhou K, Zhao Y, et al.Betulinic acid, a major therapeutic triterpene of Celastrus orbiculatus Thunb., acts as a chemosensitizer of gemcitabine by promoting Chk1 degradation.Journal of Ethnopharmacology.2023: 116295. 5. Xu X, Xie T, Zhou M, et al.Hsc70 promotes anti-tumor immunity by targeting PD-L1 for lysosomal degradation.Nature Communications.2024, 15(1): 4237. |