Suvorexant (MK-4305)

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Biological Activity

Description	Suvorexant (MK-4305) is a potent dual OX receptor antagonist with Ki of 0.55 nM and 0.35 nM for OX1 receptor and OX2 receptor, respectively. Phase 3.
Targets	OX1 receptor	OX2 receptor
IC50	0.55 nM (Ki)	0.35 nM (Ki) [1]
In vitro
In vivo	Suvorexant has excellent passive permeability and oral bioavailability, and induces favorable sleep effects in rats. [1] In addition, suvorexant significantly and dose-dependently reduces locomotor activity and promotes sleep in rats (10, 30, and 100 mg/kg), dogs (1 and 3 mg/kg), and rhesus monkeys (10 mg/kg). [2]
Features	Dual OX antagonist currently in Phase III clinical trials for the modulation of sleep.

Protocol(Only for Reference)

Kinase Assay: [1]

FLIPR Assay

For intracellular calcium measurements, Chinese hamster ovary (CHO) cells expressing the Ile408-Val variant of the orexin 1 receptor or the human orexin 2 receptor, are grown in Iscove’s modified DMEM containing 2 mM l-glutamine, 0.5 g/mL G418, 1% hypoxanthine-thymidine supplement, 100 U/mL penicillin, 100 ug/mL streptomycin, and 10% heat-inactivated fetal calf serum. The cells are seeded at 20000 cells/well into Becton-Dickinson black 384-well clear bottom sterile plates coated with poly-d-lysine. All reagents are from GIBCO-Invitrogen Corp. The seeded plates are incubated overnight at 37 °C and 6% CO2. Ala-6,12 human orexin-A as the agonist is prepared as a 0.5 mM stock solution in 1% bovine serum albumin (BSA) and diluted in assay buffer (HBSS containing 20 mM HEPES and 2.5 mM probenecid, pH 7.4) for use in the assay at a final concentration of 0.3−2 nM. Test compounds are prepared as 10 mM stock solution in DMSO, then diluted and pipetted in 384-well plates, first in DMSO, then assay buffer. On the day of the assay, cells are washed three times with 100 μL assay buffer and then incubated for 60 min (37 °C, 6% CO2) in 60 μL of assay buffer containing 1 μM Fluo-4AM ester, 0.02% pluronic acid, and 1% BSA. The dye loading solution is then aspirated and cells washed three times with 100 μL of assay buffer. Then 30 μL of that same buffer is left in each well. Within the fluorescent imaging plate reader, test compounds are added to the plate in a volume of 15 μL, incubated for 5 min, and finally 15 μL of agonist is added. Fluorescence is measured for each well at 1 s intervals for 1 min and at 6 s intervals for 4 min, and the height of each fluorescence peak is compared to the height of the fluorescence peak induced by 0.3−2 nM Ala-6,12 orexin-A with buffer in place of antagonist. For each antagonist, the IC50 value (the concentration of compound needed to inhibit 50% of the agonist response) is determined.

Animal Study: [1]

Animal Models	Adult male Sprague−Dawley rats
Dosages	~10 mg/kg
Administration	Oral gavage

1

References

Clinical Trial Information( data from http://clinicaltrials.gov)

NCT Number	Recruitment	Conditions	Sponsor /Collaborators	Start Date	Phases
NCT01293006	Completed	Insomnia	Merck Sharp & Dohme Corp.	2014-01	Phase 1
NCT01097616	Completed	Primary Insomnia	Merck Sharp & Dohme Corp.	2014-05	Phase 3

Chemical Information

Download Suvorexant (MK-4305) SDF

Chemical Name	5-Chloro-2-[(5R)-5-methyl-4-[5-methyl-2-(2H-1,2,3-triazol-2-yl)benzoyl]-1,4-diazepan-1-yl]-1,3-benzoxazole