TAPI 2

(R)-TAPI-2 is the isomer of TAPI-2 (HY-100211A). TAPI-2 (TNF Protease Inhibitor 2) is a broad-spectrum inhibitor of matrix metalloprotease (MMP), tumour necrosis factorα-converting enzyme (TACE) and a disintegrin and metalloproteinase (ADAM), with an IC50 of 20 μM for MMP[1]. TAPI-2 blocks the entry of infectious SARS-CoV[2].

Research Areas >> Cancer

Signaling Pathways >> Others >> Others

基于 hydroxamate 的金属蛋白酶抑制剂 TAPI-2 与 hmeprin 结合，hmeprin β 亚基的抑制常数 IC50 为 20±10 μM，hmeprin α 为 1.5±0.27 nM 亚基。通常，hmeprin α 比 β 亚基 受到更强烈的抑制[1]。 在不影响 ADAM17 表达的情况下，TAPI-2 会显着降低 HCP-1 和 HT29 细胞中 NICD 及其下游靶标 HES-1 的蛋白质水平。此外，在两种 CRC 细胞系中，用 TAPI-2 处理细胞可显著降低 50% 的 CSC 表型。TAPI-2 对 NICD 和 HES-1 的球形成和蛋白水平的剂量依赖性证实，所使用的浓度 (20 μM) 在 TAPI-2 的有效剂量范围 (5-40 μM) 内[3]。

TAPI-2 is dissolved in DMSO and diluted with appropriate medium before use. All experiments are performed using 20 μM TAPI-2. Cells are cultured with or without TAPI-2 for 48 hours and then seeded at 3,000 cells per well in 96-well plates. After pretreatment, increasing doses of 5-fluorouracil (5-FU) that are relevant to the recommended clinical dose (up to 2 μg/mL) are added, with or without TAPI-2, for 72 hours. Cell viability is assessed by adding MTT substrate (0.25% in phosphate-buffered saline [PBS]) in growth medium (1:5 dilution) to cells for 1 hour at 37°C. The cells are ished with PBS, and 100 μL of dimethyl sulfoxideis added. Optical density is measured at 570 nM, and relative MTT is presented as a percentage of control[3].

[1]. Wang R, et al. A Disintegrin and Metalloproteinase Domain 17 Regulates Colorectal Cancer Stem Cells and Chemosensitivity Via Notch1 Signaling. Stem Cells Transl Med. 2016 Mar;5(3):331-8.  

[2]. Kruse MN, et al. Human meprin alpha and beta homo-oligomers: cleavage of basement membrane proteins and sensitivity to metalloprotease inhibitors. Biochem J. 2004 Mar 1;378(Pt 2):383-9.  

[3]. Shiori Haga, et al. TACE Antagonists Blocking ACE2 Shedding Caused by the Spike Protein of SARS-CoV Are Candidate Antiviral Compounds. Antiviral Res. 2010 Mar;85(3):551-5.