Home Ubiquitin DUB inhibitor b-AP15

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b-AP15 DUB inhibitor

Cat.No.S4920

b-AP15 (NSC687852) is a deubiquitinases inhibitor for 19S proteasomes activity of Ub-AMC cleavage with IC50 of 2.1 μM.
b-AP15 DUB inhibitor Chemical Structure

Chemical Structure

Molecular Weight: 419.39

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Cell Culture, Treatment & Working Concentration

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
P388 cells Cytotoxicity assay Cytotoxicity against murine P388 cells, IC50=0.05 μM
human HSC2 Cytotoxicity assay Cytotoxicity against human HSC2 by MTT method, CC50=0.094 μM
HL60 cells Cytotoxicity assay Cytotoxicity against human HL60 cells in presence of RPMI1640 containing 10% fetal bovine serum by trypan blue exclusion test, CC50=0.13 μM
Molt 4/C8 cells Cytotoxicity assay Cytotoxicity against human Molt 4/C8 cells, IC50=0.15 μM
CEM cells Cytotoxicity assay Cytotoxicity against human CEM cells, IC50=0.26 μM
L1210 cells Cytotoxicity assay Cytotoxicity against murine L1210 cells, IC50=0.42 μM
HSC4 cells Cytotoxicity assay Cytotoxicity against human HSC4 cells by MTT method, CC50=0.56 μM
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Chemical Information, Storage & Stability

Molecular Weight 419.39 Formula

C22H17N3O6

 Storage (From the date of receipt)
CAS No. 1009817-63-3 Download SDF Storage of Stock Solutions

Synonyms NSC687852 Smiles C=CC(=O)N1CC(=CC2=CC=C(C=C2)[N+](=O)[O-])C(=O)C(=CC3=CC=C(C=C3)[N+](=O)[O-])C1

Solubility

In vitro
Batch:

DMSO : 48 mg/mL (114.45 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Water : Insoluble

Ethanol : Insoluble

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Mass Concentration Volume Molecular Weight

In vivo
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Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

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Mechanism of Action

Features
Not a general deubiquitinase inhibitor. Has minimal inhibition on recombinant and cytosolic nonproteasomal cysteine deubiquitinases.
Targets/IC50/Ki
USP14 [1]
UCHL5 [1]
2.1 μM
In vitro
b-AP15 inhibits the activity of two 19S regulatory-particle-associated deubiquitinases, ubiquitin C-terminal hydrolase 5 (UCHL5) and ubiquitin-specific peptidase 14 (USP14), resulting in accumulation of polyubiquitin. This compound results in a dose-dependent accumulation of the UbG76V-YFP reporter with IC50 of 0.8 μM, indicating impaired proteasome degradation. It (1 μM) results in rapid accumulation of polyubiquitinated proteins in human colon carcinoma HCT-116 cells. This chemical (2.2 μM) increases the amounts of the cyclin-dependent kinases CDKN1A and CDKNIB and the tumor suppressor TP53 in a dose-dependent manner without altering the amounts of ornithine decarboxylase 1 (ODC1) in HCT-116 cells. It (1 μM) results in G2/M phase cell-cycle arrest in HCT-116 cells, consistent with the accumulation of cell-cycle inhibitors. This compound treatment increases the number of hypodiploid cells and is associated with increased amounts of apoptotic markers, including activated caspase-3, caspase-cleaved poly-ADP ribose polymerase (PARP) and cytokeratin-18 (CK18). It is more toxic to HCT-116 cells as compared to immortalized epithelial cells (hTERT-RPE1) or peripheral blood mononuclear cells. This inhibitor inhibits deubiquitinating activity using a variety of substrates, including Ub-AMC, Ub-GFP22, ubiquitinated p53-binding protein homolog (HDM2), and K48- and K63-linked ubiquitin tetramer chains. [1] It is an inhibitor of the UPS that induced cell death via induction of the lysosomal apoptosis pathway in a cathepsin-D dependent manner. This compound elicits characteristic UPS defects including the accumulation of ubiquitin conjugates and cell cycle inhibitors such as p21, p27 and the tumor suppressor p53. It inhibits the deubiquitinase activity of both cysteine DUBs, with USP14 being slightly more sensitive than UCHL5. This chemical induces apoptosis in cells over-expressing the anti-apoptotic Bcl-2 protein and in cells lacking the p53 gene. [2] It (1 μM) inhibits ATP-induced IL-1β release from LPS-primed peritoneal macrophages. This compound (1 μM) reduces the levels of cell death induced by nigericin treatment in THP-1 cells. It (1 μM) significantly reduces the numbers of ASC specks formed after nigericin treatment in LPS-primed THP-1 cells. [3]
In vivo
b-AP15 (5 mg/kg) shows significant antitumor activity in severe combined immunodeficiency (SCID) mice with FaDu squamous carcinoma xenografts. This compound significantly delays tumor onset in mice with HCT-116 colon carcinoma xenografts. [1]
References

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