USP25/28-IN-2

USP25/28-IN-2 (compound AZ2) is a selectivity dual USP25/USP28 inhibitor with USP28 IC₅₀ values of 0.9 μM, USP28 K_a values of 0.9 μM, and USP25 IC₅₀ values of 0.88 μM. USP25/28-IN-2 modulates USP25 and USP28 activity to affect downstream pathways, modulates c-Myc oncoprotein total levels and half-life, induces apoptosis, reduces cell viability. USP25/28-IN-2 can be used for the research of colorectal carcinoma, colorectal adenocarcinoma^[1]. In Vitro:USP25/28-IN-2 (compound AZ2) (titration range) potently inhibits purified recombinant USP28 enzyme activity across multiple fluorogenic substrate assays, with IC₅₀ values ranging from 0.9 to 1.3 μM^[1].
USP25/28-IN-2 (AZ2) (titration range; 10 μM) potently inhibits purified recombinant USP25 enzyme activity in a fluorogenic substrate assay, with an IC₅₀ of 0.88 μM^[1].
USP25/28-IN-2 (AZ2) (200 μM) binds specifically and reversibly to purified recombinant USP28 protein, with a K_d of 0.9 μM as measured by ITC^[1].
USP25/28-IN-2 (AZ2) (titration range) binds specifically to purified recombinant USP28 protein, with a K_d of 10.3 μM as measured by MST^[1].
USP25/28-IN-2 (AZ2) (0.01-60 μM; 2 h) engages endogenous USP28 in HCT116 cells, with an EC₅₀ of 18.2 μM as measured by Ub-VS probe competition^[1].
USP25/28-IN-2 (AZ2) (0.01-60 μM; 2 h) engages endogenous USP25 in HCT116 cells, with an EC₅₀ of 11.5 μM as measured by Ub-VS probe competition^[1].
USP25/28-IN-2 (AZ2) (0-100 μM; 3 h) dose-dependently reduces endogenous total c-Myc protein levels in HCT116 cells after 3 h of treatment^[1].
USP25/28-IN-2 (AZ2) (0-100 μM; 3 h) dose-dependently induces apoptosis in HCT116 cells after 3 h of treatment, as measured by PARP cleavage^[1].
USP25/28-IN-2 (AZ2) (0-100 μM; 72 h) exerts dose-dependent anti-proliferative effects in multiple cancer cell lines (including HCT116, HT29, and SW480) and tissue-matched normal cell lines after 72 h of treatment, with EC₅₀ values clustered around 20 μM in cancer cells and 28 μM in normal cells^[1].