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P5091

Name: P5091
Brand: Selleck Chemicals
SKU: S7132
Rating: 5 (55 reviews)

Synonyms: P005091

Catalog No.S7132 Purity:99.91%

P5091 is a selective and potent inhibitor of ubiquitin-specific protease 7 (USP7) with EC50 of 4.2 μM and the closely related USP47.

P5091 Chemical Structure

CAS No. 882257-11-6

Purity & Quality Control

Batch: Purity: 99.91%

99.91

P5091 Related Products

Related Targets	USP/UBP UCH	Click to Expand
Related Products	PR-619 P22077 IU1 b-AP15 ML323 LDN-57444 VLX1570 EOAI3402143 PLpro inhibitor USP25/28 inhibitor AZ1 TCID ML364 SJB2-043 GSK2643943A	Click to Expand
Related Compound Libraries	Kinase Inhibitor Library FDA-approved Drug Library Natural Product Library Bioactive Compound Library-I Highly Selective Inhibitor Library	Click to Expand

Signaling Pathway

DUB Signaling Pathway

Cell Data

Cell Lines	Assay Type	Incubation Time	Activity Description	PMID
Sf9 cells	Function assay		Inhibition of recombinant USP7 expressed in Sf9 cells by Ub-CHOP reporter assay, EC50=4.2 μM	24900381
human HCT116 cells	Cytotoxic assay	72 h	Cytotoxicity against human HCT116 cells after 72 hrs, EC50=11 μM	24900381
multiple myeloma cells	Function assay		Inhibition of USP7 in multiple myeloma cells (unknown origin) by immunohistochemistry, EC50 = 4.2 μM.	25364867
Sf9	Function assay	30 mins	Inhibition of full-length recombinant USP7 (unknown origin) expressed in Sf9 cells using Ub-EKL as substrate preincubated for 30 mins followed by substrate addition by fluorescence based assay, IC50 = 4.2 μM.	28768102
Click to View More Cell Line Experimental Data

Biological Activity

Description

P5091 is a selective and potent inhibitor of ubiquitin-specific protease 7 (USP7) with EC50 of 4.2 μM and the closely related USP47.

Targets

USP7 ^[1] (Cell-free assay)	USP47 ^[2] (Cell-free assay)
4.2 μM(EC50)	4.3 μM

In vitro
In vitro	P5091 is a trisubstituted thiophene with dichlorophenylthio, nitro, and acetyl substituents mediating anti-USP7 activity. P5091 exhibits potent, specific, and selective deubiquitylating activity against USP7. In contrast, P5091 does not inhibit other DUBs or other families of cysteine proteases tested (EC50 > 100 mM). P5091 inhibits the labeling of USP7 with HA-UbVME in a concentration-dependent manner. USP7-mediated cleavage of high molecular weight polyubiquitin chains is inhibited in a dose-dependent manner by P5091. Moreover, P5091 inhibits USP7- but not USP2- or USP8-mediated cleavage of poly K48-linked ubiquitin chains. USP7 inhibition by P5091 induces HDM2 polyubiquitylation and accelerates degradation of HDM2. P5091 inhibits USP7 deubiquitylating activity, without blocking proteasome activity in MM Cells. P5091 induces a dose-dependent decrease in viability of various MM cell lines. P5091 overcomes bone marrow stromal cell-induced growth of MM Cells. P5091 decreased HDM2 and HDMX, as well as upregulated p53 and p21 levels. Overall, P5091-induced cytotoxicity is mediated in part via HDM2-p21 signaling axis and although p53 is upregulated in response to P5091 treatment, the cytotoxic activity of P5091 is not dependent on p53. ^[1]
Kinase Assay	Ubiquitin Protease Assays
Kinase Assay	Recombinant enzymes in 20 mM Tris-HCl (pH 8.0), 2 mM CaCl₂, and 2 mM β-mercaptoethanol are incubated with dose ranges of P5091 for 30 min in a 96-well plate before the addition of Ub-PLA2 and NBD C6-HPC or Ub-EKL and EKL substrate. The liberation of a fluorescent product within the linear range of the assay is monitored using a Perkin Elmer Envision fluorescence plate reader. Vehicle (2% [v/v] DMSO) and 10 mM N-ethylmaleimide (NEM) are included as controls.
Cell Research	Cell lines	293T cells
	Concentrations	10 μM
	Incubation Time	24 h
	Method	Cells were treated with indicated concentration of drug for 24 h.
Experimental Result Images	Methods	Biomarkers	Images	PMID
	Western blot	HAUSP / N-Myc USP7 / p53 / p21		29616860
	Immunofluorescence	PTEN		28418900
	Growth inhibition assay	Cell viability		30045945

In Vivo
In vivo	In animal tumor model studies, P5091 is well tolerated, inhibits tumor growth, and prolongs survival. Combining P5091 with HDAC inhibitor SAHA triggers synergistic anti-MM activity. ^[1]
Animal Research	Animal Models	CB-17 SCID-mice
	Dosages	10 mg/kg
	Administration	--

References

Chemical Information & Solubility

Molecular Weight	348.22	Formula	C₁₂H₇Cl₂NO₃S₂
CAS No.	882257-11-6	SDF	Download P5091 SDF
Smiles	CC(=O)C1=CC(=C(S1)SC2=C(C(=CC=C2)Cl)Cl)[N+](=O)[O-]
Storage (From the date of receipt)	3 years-20°Cpowder	Storage of Stock Solutions Aliquot the stock solutions to avoid repeated freeze-thaw cycles	1 year-80°Cin solvent
Storage (From the date of receipt)	3 years-20°Cpowder		1 month-20°Cin solvent

In vitro
Batch:

DMSO : 28 mg/mL ( (80.4 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Water : Insoluble

Ethanol : Insoluble

Molecular Weight Calculator

In vivo Batch: Add solvents to the product individually and in order.				In vivo Formulation Calculator
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	6.5%DMSO 1 40%PEG300 2 5%Tween80 3 48.5%ddH2O Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	1.950mg/ml (5.60mM)	Taking the 1 mL working solution as an example, add 65 μL of 30 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 485 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.

Preparing Stock Solutions

Molarity Calculator

Mass	Concentration	Volume	Molecular Weight
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Dilution Calculator Molecular Weight Calculator

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

Dosage: mg/kg Average weight of animals: g Dosing volume per animal:μL Number of animals:

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO + % + % Tween 80 + % ddH₂O

%DMSO+ %

Calculation results:

Working concentration: mg/ml;

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.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
as vortex, ultrasound or hot water bath can be used to aid dissolving.

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

Tel: +1-832-582-8158 Ext:3
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