ARN14974 (ARN-14974), a benzoxazolone carboxamide analog, is a novel and potent inhibitor of acid ceramidase with IC50 of 79 nM. It inhibits acid ceramidase activity, reduces levels of sphingosine and dihydroceramide, and increases levels of ceramide in SW403 adenocarcinoma cells and RAW 264.7 murine macrophages when used at concentrations ranging from 0.1 to 20 mM. In vivo, ARN14974 (10 mg/kg, i.v.) reduces acid ceramidase activity in multiple organs, including brain, liver, heart, lungs, and kidney, and increases pulmonary ceramide levels in mice.

Physicochemical Properties

Molecular Formula	C24H21FN2O3
Molecular Weight	404.433549642563
Exact Mass	404.153
Elemental Analysis	C, 71.27; H, 5.23; F, 4.70; N, 6.93; O, 11.87
CAS #	1644158-57-5
PubChem CID	101913546
Appearance	Off-white to pink solid powder
Density	1.3±0.1 g/cm3
Index of Refraction	1.613
LogP	5.4
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	6
Heavy Atom Count	30
Complexity	587
Defined Atom Stereocenter Count	0
SMILES	FC1C=CC(=CC=1)C1C=CC2=C(C=1)OC(N2C(NCCCCC1C=CC=CC=1)=O)=O
InChi Key	DPYVAZSLMQCVOG-UHFFFAOYSA-N
InChi Code	InChI=1S/C24H21FN2O3/c25-20-12-9-18(10-13-20)19-11-14-21-22(16-19)30-24(29)27(21)23(28)26-15-5-4-8-17-6-2-1-3-7-17/h1-3,6-7,9-14,16H,4-5,8,15H2,(H,26,28)
Chemical Name	6-(4-fluorophenyl)-2-oxo-N-(4-phenylbutyl)-3(2H)-benzoxazolecarboxamide
Synonyms	ARN-14974; ARN14974; ARN 14974; Acid Ceramidase Inhibitor 17a;
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 Note: This product requires protection from light (avoid light exposure) during transportation and storage.
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	In complicated cellular settings, ARN14974 (20 μM; 24 hours; SW403 and Raw 264.7 cells) inhibits acid ceramidase (AC), leading to the anticipated biochemical reactions of elevated ceramide and lowered sphingosine levels [1].
ln Vivo	ARN14974 (10 mg/kg; i.p.; mice) significantly lowers the activity of AC in a number of organs, including the kidneys, liver, heart, brain, and lungs [1]. After a single intraperitoneal injection of 10 mg/kg, ARN14974 (10 mg/kg) enters the bloodstream quickly in mice, with a maximum plasma concentration Cmax of 1767.9 ng/mL and a circulating half-life of 458 minutes. The Cmax and half-life of ARN14974 (1 mg/kg; intravenous) are 628 ng/mL and 72 minutes, respectively [1].
References	[1]. Benzoxazolone carboxamides: potent and systemically active inhibitors of intracellular acid ceramidase. Angew Chem Int Ed Engl. 2015;54(2):485-489.

Solubility Data

Solubility (In Vitro)

DMSO : ~50 mg/mL (~123.63 mM)

Solubility (In Vivo)

Solubility in Formulation 1: 1 mg/mL (2.47 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 10.0 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of 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: ≥ 1 mg/mL (2.47 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 10.0 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	2.4726 mL	12.3631 mL	24.7262 mL
	5 mM	0.4945 mL	2.4726 mL	4.9452 mL
	10 mM	0.2473 mL	1.2363 mL	2.4726 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.