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CB-5083 p97 AAA ATPase Inhibitor

Cat.No.S8101

CB-5083 is a potent, selective, and orally bioavailable p97 AAA ATPase inhibitor with IC50 of 11 nM. Phase 1.

Chemical Structure

Molecular Weight: 413.47

Jump to Mechanism of Action Cell Culture & Working Concentration Solubility Chemical Information, Storage & Stability Specificity

Quality Control

Batch: Purity: 99.18%

99.18

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Cell Culture, Treatment & Working Concentration

Cell Lines	Assay Type	Concentration	Incubation Time	Activity Description	PMID
A549	Function assay		6 hrs	Inhibition of p97 ATPase in human A549 cells assessed as reduction in p62 protein incubated for 6 hrs by immunofluorescence assay, IC50 = 0.49 μM.	26565666
A549	Cytotoxicity assay		72 hrs	Cytotoxicity against human A549 cells assessed as cell viability after 72 hrs by Celltiter-Glo assay, IC50 = 0.68 μM.	26565666
A549	Function assay		6 hrs	Inhibition of p97 ATPase in human A549 cells assessed as accumulation of K48 poly-ubiquitinated proteins incubated for 6 hrs by immunofluorescence assay, IC50 = 0.68 μM.	26565666
A549	Function assay		6 hrs	Inhibition of p97 ATPase in human A549 cells assessed as accumulation of CHOP poly-ubiquitinated proteins incubated for 6 hrs by immunofluorescence assay, IC50 = 1.03 μM.	26565666
Vero E6	Function assay		48 hrs	IC50 for antiviral activity against SARS-CoV-2 in the Vero E6 cell line at 48 h by immunofluorescence-based assay (detecting the viral NP protein in the nucleus of the Vero E6 cells)., IC50 = 4.89779 μM.	32353859
A549	Function assay	150 mg/kg	2 to 24 hrs	Drug uptake in plasma of mouse xenografted with human A549 cells at 150 mg/kg, po measured at 2 to 24 hrs	26565666
A549	Antitumor assay	100 mg/kg		Antitumor activity against human A549 cells xenografted in nude SCID beige mouse assessed as tumor growth inhibition at 100 mg/kg, po administered as qd4 on/3 off weekly schedule	26565666
A549	Function assay	150 mg/kg	2 to 24 hrs	Drug uptake in human A549 cells xenografted in mouse at 150 mg/kg, po measured at 2 to 24 hrs	26565666
MDA-MB231	Function assay	1 uM	8 hrs	Inhibition of p97 in human MDA-MB231 cells assessed as increase in CHOP protein levels at 1 uM after 8 hrs by Western blot analysis	28139929
HCT116	Antitumor assay	75 mg/kg	12 days	Antitumor activity against human HCT116 cells xenografted in nude mouse assessed as tumor growth inhibition at 75 mg/kg, po qd for 12 days	26565666
AMO1	Antitumor assay	100 mg/kg		Antitumor activity against human AMO1 cells xenografted in nude SCID beige mouse assessed as tumor growth inhibition at 100 mg/kg, po administered as qd4 on/3 off weekly schedule	26565666
AMO1	Antitumor assay	1 mg/kg		Antitumor activity against human AMO1 cells xenografted in nude SCID beige mouse assessed as tumor growth inhibition at 1 mg/kg, iv administered twice per week	26565666
Click to View More Cell Line Experimental Data

Chemical Information, Storage & Stability

Molecular Weight	413.47	Formula	C₂₄H₂₃N₅O₂	Storage (From the date of receipt)	3 years-20°Cpowder
CAS No.	1542705-92-9	Download SDF		Storage of Stock Solutions	1 year-80°Cin solvent 1 month-20°Cin solvent
Synonyms	N/A			Smiles	CC1=CC2=C(C=CC=C2N1C3=NC4=C(COCC4)C(=N3)NCC5=CC=CC=C5)C(=O)N

Solubility

In vitro
Batch:

DMSO : 82 mg/mL (198.32 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Ethanol : 29 mg/mL

Water : Insoluble

Molarity Calculator

Mass	Concentration	Volume	Molecular Weight
=	×	×

Dilution Calculator Molecular Weight Calculator

In vivo Batch:
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	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	4.150mg/ml (10.04mM)	Taking the 1 mL working solution as an example, add 50 μL of 83 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 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
Clear solution	5% DMSO 1 95% Corn oil Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	0.350mg/ml (0.85mM)	Taking the 1 mL working solution as an example, add 50 μL of 7 mg/ml clear DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results.
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.

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.

Mechanism of Action

Targets/IC₅₀/Ki	p97 AAA ATPase ^[1] (Cell-free assay) 11 nM
In vitro	In A549 cells, CB-5083 causes significant K48 poly-ubiquitinated protein and CHOP accumulation as well as p62 reduction, and kills tumor cells with IC50 of 680 nM. ^[1]
Kinase Assay	ATPase assay
Kinase Assay	Compounds are diluted in DMSO with a 3-fold 10-point serial dilution starting at 10 μM. The assay is done in a 384-well plate with each row as a single dilution series with duplicate of each compound concentration point. In 5 μL total volume, 20 nM p97 hexameric enzyme and 20 μM ATP are added to start the reaction. The plate is sealed and incubated at 37 °C for 15 min after mixing thoroughly in an orbital shaker. Compound dilution, ATP and enzymes addition are conducted with automated liquid handling using the Freedom Evo. ADP Glo reagents 1 and 2 are added according to the manufacturer’s protocol. The luminescence is measured by Envision plate reader as the end point of the reaction. The IC50 of each compound is derived by fitting the luminescence values to a four-parameter sigmoidal curve.
In vivo	In mice bearing human HCT 116 colon tumor xenografts, CB-5083 (75 mg/kg, p.o.) significantly inhibits tumor growth. In mice bearing established human AMO-1 multiple myeloma and A549 lung carcinoma tumor xenografts, this compound (100 mg/kg, p.o.) also results in significant tumor growth inhibition. ^[1]
References	[1] https://pubmed.ncbi.nlm.nih.gov/26565666/

Applications

Methods	Biomarkers	Images	PMID
Western blot	Bip / XBP1s / PERK / pEIF2α / CHOP p97 p-IRE1α / IRE1α / p-eIF2α / ATF4		26555175

Clinical Trial Information

(data from https://clinicaltrials.gov, updated on 2024-05-22)

NCT Number	Recruitment	Conditions	Sponsor/Collaborators	Start Date	Phases
NCT02243917	Terminated	Advanced Solid Tumors	Cleave Biosciences Inc.	October 11 2014	Phase 1
NCT02223598	Terminated	Lymphoid Hematological Malignancies\|Relapsed and Refractory Multiple Myeloma	Cleave Biosciences Inc.	August 25 2014	Phase 1

Tech Support

Handling Instructions

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

If you have any other enquiries, please leave a message.

Frequently Asked Questions

Question 1:
I am trying to administrate it to mice. Does this compound ever dissolve completely?

Answer:
It can be dissolved in 0.5% CMC Na+5% Tween 80 at 30 mg/ml as a suspension for oral gavage.

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