DUB-IN-1 is an active ubiquitin-specific proteases inhibitor (antagonist) with IC50 of 0.85 μM for USP8.

Physicochemical Properties

Molecular Formula	C20H11N5O
Molecular Weight	337.33424
Exact Mass	337.096
CAS #	924296-18-4
PubChem CID	16065490
Appearance	Light yellow to yellow solid powder
Density	1.32g/cm3
Boiling Point	632.2ºC at 760 mmHg
Flash Point	336.2ºC
Index of Refraction	1.704
LogP	3.169
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	3
Heavy Atom Count	26
Complexity	637
Defined Atom Stereocenter Count	0
SMILES	C1=CC=C(C=C1)CO/N=C/2\C3=CC=CC=C3C4=NC(=C(N=C42)C#N)C#N
InChi Key	GKOWDIBLCDZJHF-NCELDCMTSA-N
InChi Code	InChI=1S/C20H11N5O/c21-10-16-17(11-22)24-20-18(23-16)14-8-4-5-9-15(14)19(20)25-26-12-13-6-2-1-3-7-13/h1-9H,12H2/b25-19+
Chemical Name	(9E)-9-phenylmethoxyiminoindeno[1,2-b]pyrazine-2,3-dicarbonitrile
HS Tariff Code	2934.99.9001
Storage	Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

ln Vitro	DUBs-IN-1 (22 d) exhibits an IC50 of 0.85 μM against USP8, making it an effective inhibitor of ubiquitin-specific proteases. When it comes to USP7 (IC50, >100 μM), DUBs-IN-1 is inactive. DUBs-IN-1 and its derivatives, with IC50 values ranging from 0.5 μM to 1.5 μM, have the ability to decrease the viability of the HCT116 colon cancer cell line and the PC-3 prostate cancer cell line [1].
References	[1]. Synthesis and biological evaluation of 9-oxo-9H-indeno[1,2-b]pyrazine-2,3-dicarbonitrile analogues as potential inhibitors of deubiquitinating enzymes. ChemMedChem. 2010 Apr 6;5(4):552-8.

Solubility Data

Solubility (In Vitro)

DMSO : ~100 mg/mL (~296.45 mM)

Solubility (In Vivo)

Solubility in Formulation 1: 2.5 mg/mL (7.41 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (7.41 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.9645 mL	14.8223 mL	29.6446 mL
	5 mM	0.5929 mL	2.9645 mL	5.9289 mL
	10 mM	0.2964 mL	1.4822 mL	2.9645 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.