ACT-462206 is an orally bioactive dual Orexin 1/Orexin 2 receptor blocker (antagonist) with IC50s of 60 nM (Orexin 1) and 11 nM (Orexin 2), respectively. ACT-462206 penetrates the BBB (blood-brain barrier) and may be utilized in the research on insomnia, stress/anxiety-related disorders, and addiction.

Physicochemical Properties

Molecular Formula	C20H24N2O4S
Molecular Weight	388.480564117432
Exact Mass	388.146
CAS #	1361321-96-1
PubChem CID	40924317
Appearance	White to off-white solid powder
LogP	4.195
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	5
Heavy Atom Count	27
Complexity	600
Defined Atom Stereocenter Count	1
SMILES	CC1=CC(=CC(=C1)NC(=O)[C@@H]2CCCN2S(=O)(=O)C3=CC=C(C=C3)OC)C
InChi Key	NHPQGZOBHSVTAQ-IBGZPJMESA-N
InChi Code	InChI=1S/C20H24N2O4S/c1-14-11-15(2)13-16(12-14)21-20(23)19-5-4-10-22(19)27(24,25)18-8-6-17(26-3)7-9-18/h6-9,11-13,19H,4-5,10H2,1-3H3,(H,21,23)/t19-/m0/s1
Chemical Name	(2S)-N-(3,5-dimethylphenyl)-1-(4-methoxyphenyl)sulfonylpyrrolidine-2-carboxamide
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	Orexin receptor type 1 (OX1) and type 2 (OX2) are two closely related G protein-coupled receptors (GPCRs) to which orexin can bind after being released in an axon terminal in a Ca2+-sensitive manner[1]. 17 nM (hOX1), 2.4 nM (hOX2), 28 nM (rOX1), 9.9 nM (rOX2), 27 nM (dOX1), and 4.2 nM (dOX2) are the binding affinities of ACT-462206 and Kb [1]. With IC50s of 60 nM (hOX1), 11 nM (hOX2), 48 nM (rOX1), 9.6 nM (rOX2), 68 nM (dOX1), and 26 nM (dOX2), ACT-462206 suppresses Orexin activity [1]. CYP450 3A4T and 3A4M are inhibited by ACT-462206, with IC50 values of 15 μM and 29 μM, respectively[1].
ln Vivo	ACT-462206 (Compound 24) (100 mg/kg; oral; 3-hour sampling) has concentrations in the brain and plasma of male Wistar rats of 1219 ng/g and 2267 ng/mL, respectively, indicating that it can cross the blood-brain barrier [1]. In male Wistar rats and male Beagle dogs, ACT-462206 (10-300 mg/kg; oral; single dose) reduces arousal and increases non-rapid eye movement (NREM) and rapid eye movement (REM) sleep. 1]. The oral medication ACT-462206 (100, 300 mg/kg; single dose) has anxiolytic-like effects on rats, improving the startle reflex, decreasing movement, body temperature, and heart rate increases brought on by social stress [1]. Different species' pharmacokinetics [1]: Route dose (mg/kg), AUC (ng·h/mL), CL (mL/min/kg), Vss (L/kg), t1/2 (h), cmax (ng/mL), tmax (h), F1/2 (%) Rat iv 1 586 29 1.8 1.9 / / / po 10 2310 / / / 1600 0.5 39 Dog iv 1 1490 11 1.4 1.7 / / / po 3 2750 / / / 426 0.5 52
Animal Protocol	Animal/Disease Models: Male Wistar rat[1] Doses: 0, 10, 30, 100, 300 mg/kg Route of Administration: po (oral gavage); single dose Experimental Results: shortened the first sustained non-rapid eye movement sleep (60 seconds) and latency to first sustained REM sleep (30 seconds). Total arousal time and behavioral home cage activity were dose-dependently diminished (one-way ANOVA; p < 0.001), while REM and non-REM sleep time were increased.
References	[1]. Structure-activity relationship, biological, and pharmacological characterization of the proline sulfonamide ACT-462206: a potent, brain-penetrant dual orexin 1/orexin 2 receptor antagonist. ChemMedChem. 2014 Nov;9(11):2486-96.

Solubility Data

Solubility (In Vitro)

DMSO : ≥ 100 mg/mL (~257.41 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.44 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 (6.44 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	2.5741 mL	12.8707 mL	25.7414 mL
	5 mM	0.5148 mL	2.5741 mL	5.1483 mL
	10 mM	0.2574 mL	1.2871 mL	2.5741 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.