HJC0350 is a potent and selective antagonist/inhibitor of EPAC2 with IC50 of 0.3 μM, it exhibited no inhibition on Epac1. Under both physiological and pathological conditions, the isoforms of exchange proteins directly activated by cAMP (EPAC), EPAC1 and EPAC2, react to the second messenger cAMP and control a host of intracellular functions. HJC 0350 competed with 8-NBD-cAMP in binding recombinant fusion protein EPAC2 with IC50 value of 0.3 μM and exhibited 133-fold more potent than cAMP, which competed with 8-NBD-cAMP in binding EPAC2 with IC50 value of 40 μM. HJC 0350 (25 μM) demonstrated EPAC2-specific antagonist properties as it inhibited EPAC2 GEF activity but had no effect on EPAC1-mediated Rap1-GDP exchange activity or cAMP-mediated PKA activation.

Physicochemical Properties

Molecular Formula	C15H19NO2S
Molecular Weight	277.38
Exact Mass	277.114
Elemental Analysis	C, 64.95; H, 6.90; N, 5.05; O, 11.54; S, 11.56
CAS #	885434-70-8
Related CAS #	885434-70-8
PubChem CID	22200891
Appearance	Solid powder
LogP	4.347
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	2
Rotatable Bond Count	2
Heavy Atom Count	19
Complexity	402
Defined Atom Stereocenter Count	0
SMILES	O=S(N1C(C)=CC(C)=C1)(C1C(C)=CC(C)=CC=1C)=O
InChi Key	AFZWZVLPIMHLSE-UHFFFAOYSA-N
InChi Code	InChI=1S/C15H19NO2S/c1-10-6-12(3)15(13(4)7-10)19(17,18)16-9-11(2)8-14(16)5/h6-9H,1-5H3
Chemical Name	2,4-dimethyl-1-(2,4,6-trimethylphenyl)sulfonylpyrrole
Synonyms	HJC 0350; HJC-0350; HJC0350
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

Targets	EPAC2 ( IC50 = 0.3 µM )
ln Vitro	In vitro activity: HJC0350 is approximately 133 times more potent than cAMP and has an apparent IC50 value of 0.3 µM for competing with 8-NBD-cAMP binding of EPAC2. It is discovered that HJC0350 is an EPAC2-specific antagonist since it does not impede EPAC1-mediated Rap1-GDP exchange activity at 25 µM in the presence of equal concentrations of cAMP. When HEK293/EPAC2-FL cells are pretreated with 10 µM HJC0350, the 007-AM-induced decrease in FRET is completely blocked[1].
ln Vivo
Enzyme Assay	HJC 0350 is a potent and selective antagonist of EPAC2. HJC 0350 competed with 8-NBD-cAMP in binding recombinant fusion protein EPAC2 with IC50 value of 0.3 μM and exhibited 133-fold more potent than cAMP, which competed with 8-NBD-cAMP in binding EPAC2 with IC50 value of 40 μM. HJC 0350 (25 μM) inhibited EPAC2 GEF activity in the presence of 25 μM cAMP, but did not affect cAMP-mediated PKA activation or EPAC1-mediated Rap1-GDP exchange activity. This suggests that HJC 0350 is an EPAC2-specific antagonist.
Cell Assay	HJC 0350 (10 μM) completely inhibited the decrease of FRET in HEK293/EPAC2-FL cells induced by 007-AM (a membrane permeable EPAC selective cAMP analogue) in HEK293 cells expressing EPAC1- or EPAC2-based fluorescence resonance energy transfer (FRET) sensor (EPAC2-FL or EPAC1-FL). However, it had no effect on HEK293/EPAC1-FL cells.
Animal Protocol
References	[1]. Identification and characterization of small molecules as potent and specific EPAC2 antagonists. J Med Chem. 2013 Feb 14;56(3):952-62.

Solubility Data

Solubility (In Vitro)

DMSO: 33.3~52 mg/mL (120.2~187.5 mM) Water: <1 mg/mL Ethanol: <1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (9.01 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	3.6052 mL	18.0258 mL	36.0516 mL
	5 mM	0.7210 mL	3.6052 mL	7.2103 mL
	10 mM	0.3605 mL	1.8026 mL	3.6052 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.