PR619 是一种DUB抑制剂，可诱导内质网应激和内质网应激相关的凋亡，对USP4、USP8、USP7、USP2和USP5的EC50分别为 3.93、4.9、6.86、7.2 和 8.61 μM。

PR-619 is a non-selective, reversible inhibitor of the deubiquitinylating enzymes (DUBs).

PR-619以剂量和时间依赖性方式（20至150 μM,0.5至20 h）在HEK293T细胞中增加总体蛋白质多聚泛素化。PR619处理导致K48和K63连接的polyUb链上调。PR-619诱导HCT116细胞死亡,EC50为6.3 μM。

Ub-PLA2 assay: Recombinant enzymes in 20 mM Tris-HCl, pH 8.0, 2 mM CaCl2 and 2 mM β-mercaptoethanol (DUB assay buffer) are preincubated with single doses or dose ranges of PR-619 or P22077 for 30 minutes in a 96 well plate before the addition of Ub-PLA2 and NBD C6-HPC. The liberation of a fluorescent product within the linear range of the assay is monitored at room temperature using a fluorescence plate reader. Vehicle (2%(v/v) DMSO) and 10 mM N-ethylmaleimide are included as controls. Where ≥60% inhibition is observed, EC50 values are determined using a sigmoidal dose response equation.

72 h hours later, 0.2 mg/mL resazurin prepared in phosphate-buffered saline is added to each well and the cells are incubated for an additional 3-6 h. The fluorescence of the resazurin reduction product is measured using Ex=535 nm and Em=590 nm filters on a fluorimeter. The EC50 values are calculated in Prism.(Only for Reference)

2645-32-1

C7H5N5S2

223.33

2,6-Diamino-3,5-dithiocyanopyridine;PR 619

DMSO:201.5 mM
Powder: -20°C for 3 years
In solvent: -80°C for 2 years