CID-2858522 is a potent and specific antigen receptor-mediated NF-κB inhibitor (antagonist) with IC50 of 70 nM.

Physicochemical Properties

Molecular Formula	C28H39N3O3
Molecular Weight	465.638
Exact Mass	465.299
CAS #	758679-97-9
PubChem CID	2858523
Appearance	Off-white to light yellow solid powder
Density	1.1±0.1 g/cm3
Boiling Point	618.9±65.0 °C at 760 mmHg
Flash Point	328.1±34.3 °C
Vapour Pressure	0.0±1.9 mmHg at 25°C
Index of Refraction	1.573
LogP	6.64
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	9
Heavy Atom Count	34
Complexity	661
Defined Atom Stereocenter Count	0
InChi Key	CYCGGKILBWERDJ-UHFFFAOYSA-N
InChi Code	InChI=1S/C28H39N3O3/c1-17-12-22-23(13-18(17)2)31(26(30-22)29-10-9-11-32)16-24(33)19-14-20(27(3,4)5)25(34)21(15-19)28(6,7)8/h12-15,32,34H,9-11,16H2,1-8H3,(H,29,30)
Chemical Name	1-(3,5-ditert-butyl-4-hydroxyphenyl)-2-[2-(3-hydroxypropylamino)-5,6-dimethylbenzimidazol-1-yl]ethanone
Synonyms	CID 2858522; CID-2858522; CID2858522
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	Compound 1, CID-2858522, has an IC50 of 70 nM, which inhibits NF-κB mediated by antigen receptors. With an IC50 of 85 μM, CID-2858522 also inhibits testosterone hydroxylase when human liver microsomes (HLM) and a NADPH-generating apparatus are present [1]. CID-2858522 inhibits NF-κB reporter gene activity in the HEK293 cell line utilized for primary screening in a concentration-dependent manner with an IC50 of approximately 70 nM and a maximal inhibition of 0.25–0.5 μM. On the other hand, CID-2858522 exhibits selectivity for the NF-κB pathway activated by PMA/Ionomycin because it does not suppress TNF-induced NF-κB-reporter gene activity at concentrations as high as 4 μM. At doses ≤8 μM, CID-2858522 is not hazardous to HEK293 cells, according to cell viability tests. In transient transfection studies, CID-2858522 also effectively suppresses NF-κB reporter gene activity caused by PMA/Ionomycin[2].
ln Vivo	Using a small cohort of three male mice, in vivo dose-exposure profiling of CID-2858522 (Compound 1a) is carried out. When evaluated at t=3 h, CID-2858522 displays usual dose-dependent behavior, but at 0.5 h, serum levels for the 30 mg/kg dose are higher than for the 50 mg/kg dose. This indicates nonlinear pharmacokinetics. One possible explanation for the increased accumulation observed at a dose of 50 mg/kg is a depot effect brought on by CYP3A4 inhibition. At t=3 h, the cohort shows definite symptoms of morbidity at the 50 mg/kg dose[2].
References	[1]. Selective benzimidazole inhibitors of the antigen receptor-mediated NF-kappaB activationpathway. Bioorg Med Chem. 2010 Mar 1;18(5):1918-24. [2]. Inhibition of protein kinase C-driven nuclear factor-kappaB activation: synthesis, structure-activity relationship, and pharmacological profiling of pathway specific benzimidazole probe molecules. J Med Chem. 2010 Jun 24;53(12):4793-. [3]. Chemical biology strategy reveals pathway-selective inhibitor of NF-kappaB activation induced by protein kinase C. ACS Chem Biol. 2010 Mar 19;5(3):287-99.
Additional Infomation	1-(3,5-ditert-butyl-4-hydroxyphenyl)-2-[2-(3-hydroxypropylamino)-5,6-dimethyl-1-benzimidazolyl]ethanone is an aromatic ketone.

Solubility Data

Solubility (In Vitro)

DMSO : ~25 mg/mL (~53.69 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.37 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (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 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 (5.37 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. 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 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.1476 mL	10.7379 mL	21.4758 mL
	5 mM	0.4295 mL	2.1476 mL	4.2952 mL
	10 mM	0.2148 mL	1.0738 mL	2.1476 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.