This is a DEA controlled substance schedule IV Suvorexant (also known as 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. Suvorexant is a medication that Merck is developing to treat insomnia. Phase III trials are presently being conducted on it. Suvorexant functions by inhibiting wakefulness as opposed to promoting sleep. Suvorexant (MK-4305), a dual orexin receptor (OXR) antagonist (DORA), has demonstrated potential in the treatment of insomnia and sleep disorders.

Physicochemical Properties

Molecular Formula	C23H23CLN6O2
Molecular Weight	450.9207
Exact Mass	450.16
CAS #	1030377-33-3
Related CAS #	1030377-33-3
PubChem CID	24965990
Appearance	Typically exists as solid at room temperature
LogP	4.9
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	3
Heavy Atom Count	32
Complexity	664
Defined Atom Stereocenter Count	1
SMILES	C[C@@H]1CCN(CCN1C(=O)C2=C(C=CC(=C2)C)N3N=CC=N3)C4=NC5=C(O4)C=CC(=C5)Cl
InChi Key	JYTNQNCOQXFQPK-MRXNPFEDSA-N
InChi Code	InChI=1S/C23H23ClN6O2/c1-15-3-5-20(30-25-8-9-26-30)18(13-15)22(31)29-12-11-28(10-7-16(29)2)23-27-19-14-17(24)4-6-21(19)32-23/h3-6,8-9,13-14,16H,7,10-12H2,1-2H3/t16-/m1/s1
Chemical Name	[(7R)-4-(5-chloro-1,3-benzoxazol-2-yl)-7-methyl-1,4-diazepan-1-yl]-[5-methyl-2-(triazol-2-yl)phenyl]methanone
Synonyms	MK4305; MK 4305; 1030377-33-3; BELSOMRA; Suvorexant (MK-4305); UNII-081L192FO9; MK-4305
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	orexin receptor/OX
ln Vitro	In vitro activity: Suvorexant, also referred to as MK-4305, is a strong antagonist of both OX1 and OX2 receptors, with Ki values of 0.55 nM and 0.35 nM, respectively. Merck is the company that developed suvorexant to treat insomnia. Phase III trials are presently being conducted on it. Suvorexant functions by inhibiting wakefulness as opposed to promoting sleep. Suvorexant (MK-4305), a dual orexin receptor (OXR) antagonist (DORA), has demonstrated potential in the treatment of insomnia and sleep disorders.
ln Vivo	Suvorexant (25 mg/kg) was tested in mice during the inactive phase (lights on), when sleep is more common and orexin levels are typically low, in an in-vivo study. Suvorexant was found to significantly alter the architecture of sleep by increasing REM specifically during the first four hours after dosage. Suvorexant only considerably reduced wake at the tested doses for the first hour, while IPSU had no effect on wake time. These findings imply that, in contrast to DORAs, OX2R preferring antagonists may have a reduced propensity to disturb NREM/REM architecture.
Enzyme Assay	MK-4305 possesses a Ki value of 0.55 nM for OX1 receptor and 0.35 nM for OX2 receptor, making it a strong antagonist of both receptors. Bioactivation Assay[1] Human liver microsomes (pooled, BD Gentest) were preincubated at 1 mg/mL protein in 100 mM potassium phosphate buffer (pH 7.4) with 10 μM test compound, 1 mM MgCl2, 1 mM EDTA, 5 mM glutathione, and 1 mM NADPH for 60 min at 37 °C. The reactions were terminated with 25% acetonitrile containing 0.15 μM labetalol (internal standard). The samples were vortex-mixed and centrifuged at 14000 rpm for 10 min. The supernatant from each sample was transferred to an HPLC vial for HRMS analysis. UPLC-high resolution mass spectrometry (HRMS) was used to identify the GSH-derived adducts. The system consisted of a Waters Acquity sample manager and two Waters Acquity UPLC pumps. HRMS was performed using a Waters Q-TOF Xevo mass spectrometer. Separation was achieved using a Phenomenex Synergi 2.5 μm MAX-RP 100 Å column (50 mm × 2 mm) heated to 60 °C. The mobile phase consisted of water with 0.1% formic acid (solvent A) and acetonitrile with 0.1% formic acid (solvent B) at a flow rate of 0.5 mL/min. The gradient began with 5% solvent B for the first minute, followed by a linear increase to 15% solvent B over the next 0.5 min. Solvent B was then increased to 50% over the next 11.5 min followed by a further increase to 90% over 2 min. The column was then washed with 95% solvent B for 1.5 min. At the end of each run, the column was re-equilibrated for 5 min at the initial conditions. Mass spectral analyses were carried out using electrospray ionization in positive ion mode. The ESI capillary voltage was set at 1.5 kV, and the source and desolvation temperatures were set at 100 and 600 °C, respectively. The mass scan range was 150 to 1000 amu with 0.25 s/scan. The lock mass of 588.8691 amu was used with a frequency of one scan per 5 scans. The relative amount of GSH adducts formed with each test compound was estimated using peak area ratios. The areas obtained from the mass spectral peaks associated with GSH-derived adducts were divided by the area of the internal standard, labetalol. Radioligand Binding Assay[1] Membranes were prepared from expressed human orexin 2 receptor (hOX2R) and the Ile408-Val variant of orexin 1 receptor (hOX1R) in CHO cells according to the method described by Kunapuli et al. Confluent CHO/OX2R and CHO/OX1R cells were dissociated from flasks with PBS/1 mM EDTA and centrifuged at 1000g for 10 min. The cell pellets were homogenized with a Polytron in ice-cold 20 mM Hepes, 1 mM EDTA, at pH 7.4, and centrifuged at 20000g for 20 min at 4 °C. This process was repeated twice. The final membrane pellet was resuspended at 5 mg of membrane protein/mL in assay buffer (20 mM Hepes, 125 mM NaCl, 5 mM KCl, pH 7.4). Bovine serum albumin was added to achieve a final concentration of 1% and aliquots stored at −80 °C. Radioligand binding assays were performed utilizing an automated Tecan Liquid handling system and Packard Unifilter-96 as described by Mosser et al. Assays were performed at room temperature in 96-well microtiter plates with a final assay volume of 1.0 mL in 20 mM Hepes buffer (pH 7.4) containing 125 nM NaCl and 5 mM KCl. Solutions of test compounds were prepared in DMSO and serially diluted with DMSO to yield 20 μL of each of 10 solutions differing by 3-fold in concentration. Nonspecific binding (NSB) is determined using a high-affinity ligand (1 μM final concentration) and total binding (TB) is determined by using DMSO (2% final concentrations). A solution of receptor (30 pM final, typically 2−10 μg membranes), and tritiated ligands (∼80 Ci/mmole) were added to the test compounds. For the OX2R receptor, 0.15 nM of compound 18 (KD = 0.3 nM) was used. For the OX1R receptor, 0.7 nM of compound 19 (KD = 3 nM) was used. The OX1R assay was also performed with equivalent results using compound 20 at a concentration of 0.03 nM (KD = 0.03 nM); however, in this case 920 μL of membranes was added first to the compounds followed by the addition of 60 μL of the hot ligand. After 3 h of incubation at room temperature (20 h for compound 20), samples were filtered through Packard GF/B filters (presoaked in 0.2% PEI, polyethyleninine Sigma P-3143) and washed five times with 1 mL of cold 20 mM Hepes buffer (pH 7.4) per wash. After vacuum drying of the filter plates, 50 μL of Packard Microscint-20 was added and bound radioactivity (CPM bound) determined in a Packard TopCount. Radioligand binding[2] Cell membranes from HEK293 cells transiently expressing the human OX2 receptor (Supporting Information) were incubated with [3H]-EMPA in Krebs assay buffer (8.5 mM HEPES, 1.3 mM CaCl2, 1.2 mM MgSO4, 118 mM NaCl, 4.7 mM KCl, 4 mM NaHCO3, 1.2 mM KH2PO4, 11 mM glucose, pH 7.4) in a total assay volume of 0.25 mL with a final DMSO concentration of 1%. After 90 min incubation at room temperature, the reaction was terminated by rapid filtration through GF/B 96-well glass fibre plates with 5 × 0.25 mL washes with ddH2O using a Tomtec cell harvester. Bound radioactivity was determined through liquid scintillation using Lablogic SafeScint and detected on a microbeta liquid scintillation counter. Non-specific binding was determined as that remaining in the presence of a 10 μM saturating concentration of the antagonist EMPA. Saturation studies were carried out by incubating membranes (2 μg protein/well) with a range of concentrations of [3H]-EMPA (0.4 nM–15 nM). Radioligand concentrations were determined using SafeScint and a Beckman LS 6000 liquid scintillation counter. Competition binding was performed incubating membranes (2 μg protein/well) with 1.5 nM concentration of [3H]-EMPA and a range of concentrations of the test compound such as 3 (Suvorexant / MK-4305). Association kinetics for the radioligand were determined by adding the same cell membrane (2 μg protein/well) to wells containing Krebs buffer with 1% DMSO and 1.5 nM radioligand at various time points up to a total of 3 h. Dissociation kinetics were determined by pre-equilibrating membranes and [3H]-EMPA for 90 min; a saturating concentration of cold EMPA (100 μM) was then added at various time points to prevent re-association of the radioligand as it dissociates from the receptor.
Cell Assay	Based on an MDS Pharma off-target screen of 170 enzymes, receptors, and ion channels, an in vitro investigation revealed that MK-4305 had a clean ancillary profile (>10000-fold selectivity for OX2R). FLIPR Assay[1] 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, were 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 were seeded at 20000 cells/well into Becton-Dickinson black 384-well clear bottom sterile plates coated with poly-d-lysine. All reagents were from GIBCO-Invitrogen Corp. The seeded plates were incubated overnight at 37 °C and 6% CO2. Ala-6,12 human orexin-A as the agonist was 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 were 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 were 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 was 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 were added to the plate in a volume of 15 μL, incubated for 5 min, and finally 15 μL of agonist was added. Fluorescence was 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 was 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) was determined.[1] Functional inositol phosphate and ERK1/2 phosphorylation assays[2] Cell-based inositol phosphate and ERK1/2 phosphorylation functional assays were performed in 96-well plates 24 h after seeding with CHO cells stably expressing the human orexin-2 receptor at a density of 25 000 cells/well; full assay details are in the Supporting Information.
Animal Protocol	Rat Sleep Assay[1] Adult male Sprague−Dawley rats (450−600 g; Taconic Farms, Germantown, NY) were subcutaneously implanted with telemetric physiologic monitors (model F50-EEE or 4ET SI; Data Sciences International, Arden Hills, MN) that were used to simultaneously record both the electrocorticogram (ECoG) and electromyogram (EMG) activities of the rat. For placement of the 4ET SI, animals were anesthetized with isoflurane and electrodes for recording ECoG signals and EMG signals were placed. Position of the wires are based on the following coordinates. Channel 1 wire. From Lambda AP +2, ML +2 −2. Channel 2 wires From BREGMA AP +1.5 ML +3.2 (hole 1) AP −10.5 (hole2). Channel 3 wires From BREGMA AP −3.0 ML +1.5, −3.5. EMG lead placement was in neck muscle. An incision was made ∼3−5 cm in length midline on the dorsal thorax to form a pocket on the left and right side of midline, and the telemetry module was placed with a saddlebag placement method. The animals were given a single dose of antibiotic (gentomycin, 5.8 mg/kg) and an analgesic (buprenorphine, 0.1 mL) within 3 h following surgery. The animals were allowed to recover from surgery for at least two weeks prior to recording. Throughout these experiments, animals were housed individually in plastic cages (19 in. × 101/2 in. × 8 in.; Lab Products, Seaford, DE) and were provided water and food ad libitum. Lights were on a 12 h light: 12 h dark cycle with lights off at 4:00 a.m. and on at 4:00 p.m. ECoG and EMG signals were collected simultaneously from all animals using Dataquest ART software system, digitally sampled at 500 Hz, and stored on a PC for off-line analysis. The hydrochloride salt of compound 10 (458 mg) was dissolved in 70.2 mL of a 20% aqueous solution of TPGS and administered by oral gavage at 10 mpk of the free-base equivalent to four rats, 5 h into their active period (09:00 or ZT 17:00). For 3 (Suvorexant / MK-4305), the free-base (1.27 g) was suspended in 70.2 mL of a 20% aqueous solution of TPGS and dosed as above. Recordings were started just prior to compound administration and were collected for 23 h. The experiments were based on a standard crossover design with two animals receiving compound for one week and the complementary group receiving vehicle, followed by a week of reversed administration. All animals were exposed to two days administration of orally gavaged vehicle prior to initiation of experimental drug administration to allow for habituation. For baseline sleep measurements, continuous recordings were collected for two days to get average sleep behaviors for each animal over contiguous days prior to drug and vehicle administration. During the drug administration studies, recordings were collected each day prior to, during, and following drug administration. Recordings were begun prior to compound administration so that the exact time of administration was recorded within the raw data file as artifactual noise which was caused by removing the implanted transmitter from the receptive field of the receiver during administration. This information allowed a direct measure of drug/vehicle administration time during offline analysis and was not included in the data analysis. Following the completion of data collection, all data were scored with automated sleep stage analysis software, Somnologica. Assignment of sleep stages was made in general accord with those described by J. M. Monti’s group.Sleep/wake stages were assigned based upon a combination of level of movement within the field of the radio frequency receiver over which individually housed rats were caged, EMG activity, and ECoG frequencies over 10 s epochs. Active wake was assigned to the epoch when movement of the animal was detected over the receiver or when there was an active EMG signal over the epoch and the ECoG frequencies consisted of low-voltage high frequency activity. An epoch was scored as light sleep when there was no movement activity, the EMG was moderately activ,e and the ECoG consisted of either theta or theta activity mixed with less than 50% of the epoch showing delta activity. Delta sleep was scored when there was no gross movement, reduced EMG activity, and the ECoG consisted of more than 50% delta wave activity (i.e., 0.5 to 4 Hz). Rapid eye movement (REM) sleep was scored when there was no movement or EMG activity and the ECoG consisted of primarily theta activity. Results of staging were grouped into 30 min periods following drug administration and the number of entries into each stage and the duration of minutes spent in each stage were calculated. The results for all four animals were averaged by treatment, or vehicle, over seven administration nights and the results were statistically compared based upon a mixed ANOVA analysis. Ex Vivo Occupancy Assay[1] Transgenic rats expressing human OX2R were dosed intravenously by infusion over a 30 min period or orally with 3 (Suvorexant / MK-4305) at doses of 0.1−2.0 mg/kg in 25% hydroxypropyl-β-cyclodextrin and then sacrificed. Samples of brain were quickly removed and frozen for use in the ex vivo occupancy assay, while a second set of tissue samples, a plasma sample, and CSF were frozen for LCMS determination of drug levels. For the ex vivo assay, approximately 60 mg of cord or brain was homogenized in 67 volumes of ice-cold assay buffer (20 mM HEPES, 120 mM NaCl, 5 mM KCl, pH7.4) and centrifuged at 21000g for 1 min. The pellets were resuspended in ice-cold buffer at a concentration of 10 mg tissue/mL and 100 μL aliquots were rapidly distributed to tubes with 0.5 mL rof oom temperature buffer containing 200 pM compound A. At 2, 4, 6, 8, 10, 12, and 15 min following membrane addition, incubations were terminated by filtration of three tubes over glass fiber filters. A parallel set of incubations performed in the presence of 1 μM of an unlabeled, potent DORA (OX2R Ki = 1.0 nM) was used to determine nonspecific radioligand binding at each time point. Radioactivity on the filters was determined by liquid scintillation counting and compound A rates of association were determined by linear regression. Receptor occupancy in a drug treated animal is calculated as: % occupancy = (1 − (slopedrug/slopevehicle)) × 100. The concentrations of drug required to achieve 90% receptor occupancy were derived by nonlinear curve fitting using Prism software. N/A Mice
ADME/Pharmacokinetics	Absorption, Distribution and Excretion Peak concentrations occur at a median Tmax of 2 hours under fasted conditions. Ingestion of suvorexant with a high-fat meal has no effect on AUC or Cmax, but may delay Tmax by approximately 1.5 hours. Mean absolute bioavailability of 10 mg is 82%. Approximately 66% is eliminated in feces and 23% is eliminated in urine. Mean volume of distribution is approximately 49 litres. Metabolism / Metabolites Suvorexant is primarily metabolized by cytochrome-P450 3A4 enzyme (CYP3A4) with a minor contribution from CYP2C19. Major circulating metabolites are suvorexant and a hydroxy-suvorexant metabolite, which is not expected to be pharmacologically active. There is potential for drug-drug interactions with drugs that inhibit or induce CYP3A4 activity. Biological Half-Life Mean half life is approximately 12 hours.
Toxicity/Toxicokinetics	Hepatotoxicity In several clinical trials, suvorexant was found to be well tolerated, with serum ALT elevations in 0 to 5% of patients, usually with higher doses, and resolving spontaneously without dose modification. In the registration trials of suvorexant, there were no reports of clinically apparent liver injury. Suvorexant has been available for a limited period of time, but has yet to be implicated in causing clinically apparent liver injury even with an overdose. Likelihood score: E (unlikely cause of clinically apparent liver injury). Effects During Pregnancy and Lactation ◉ Summary of Use during Lactation Data from two women indicate that amounts of suvorexant in milk are very low. If suvorexant is required by the mother, it is not a reason to discontinue breastfeeding. If suvorexant is used, monitor the infant for sedation, especially if the infant is a newborn or preterm. Until more data become available, an alternate drug may be preferred, especially while nursing a newborn or preterm infant. ◉ Effects in Breastfed Infants Relevant published information was not found as of the revision date. ◉ Effects on Lactation and Breastmilk Relevant published information was not found as of the revision date. Protein Binding Suvorexant is extensively bound (>99%) to human plasma proteins and does not preferentially distribute into red blood cells. It binds to both human serum albumin and alpha1-acid glycoprotein.
References	[1]. J Med Chem . 2010 Jul 22;53(14):5320-32. [2]. Br J Pharmacol . 2014 Jan;171(2):351-63.
Additional Infomation	Suvorexant is an aromatic amide obtained by formal condensation of the carboxy group of 5-methyl-2-(2H-1,2,3-triazol-2-yl)benzoic acid with the secondary amino group of 5-chloro-2-[(5R)-5-methyl-1,4-diazepan-1-yl]-1,3-benzoxazole. An orexin receptor antagonist used for the management of insomnia. It has a role as a central nervous system depressant and an orexin receptor antagonist. It is a member of 1,3-benzoxazoles, a member of triazoles, a diazepine, an aromatic amide and an organochlorine compound. Suvorexant is a DEA Schedule IV controlled substance. Substances in the DEA Schedule IV have a low potential for abuse relative to substances in Schedule III. It is a Depressants substance. Suvorexant is a selective dual antagonist of orexin receptors OX1R and OX2R that promotes sleep by reducing wakefulness and arousal. It has been approved for the treatment of insomnia. Suvorexant is an Orexin Receptor Antagonist. The mechanism of action of suvorexant is as an Orexin Receptor Antagonist, and P-Glycoprotein Inhibitor, and Cytochrome P450 3A Inhibitor. Suvorexant is an orexin receptor antagonist used for the treatment of insomnia and sleep disorders. Suvorexant therapy is associated with rare occurrence of transient serum enzyme elevations, but has not been implicated in cases of clinically apparent liver injury. Suvorexant is an orally bioavailable antagonist of the orexin receptors orexin receptor type 1 (OX1R) and orexin receptor type 2 (OX2R), that can be used for the treatment of insomnia. Upon oral administration, suvorexant targets and binds to the orexin receptors OX1R and OX2R. This blocks the binding of the neuropeptides orexin-A and orexin-B to OX1R and OX2R, and prevents wakefulness that results from orexin signaling. Drug Indication Suvorexant is indicated for the treatment of insomnia characterized by difficulties with sleep onset and/or sleep maintenance. FDA Label Mechanism of Action Suvorexant is a dual antagonist of orexin receptors OX1R and OX2R. It exerts its pharmacological effect by inhibiting binding of neuropeptides orexin A and B, also known as hypocretin 1 and 2, that are produced by neurons in the lateral hypothalamus. These neurons control the wake-promoting centers of the brain and are active during wakefulness, especially during motor activities, and stop firing during sleep. By inhibiting the reinforcement of arousal systems, suvorexant use causes a decrease in arousal and wakefulness, rather than having a direct sleep-promoting effect. Despite increased understanding of the biological basis for sleep control in the brain, few novel mechanisms for the treatment of insomnia have been identified in recent years. One notable exception is inhibition of the excitatory neuropeptides orexins A and B by design of orexin receptor antagonists. Herein, we describe how efforts to understand the origin of poor oral pharmacokinetics in a leading HTS-derived diazepane orexin receptor antagonist led to the identification of compound 10 with a 7-methyl substitution on the diazepane core. Though 10 displayed good potency, improved pharmacokinetics, and excellent in vivo efficacy, it formed reactive metabolites in microsomal incubations. A mechanistic hypothesis coupled with an in vitro assay to assess bioactivation led to replacement of the fluoroquinazoline ring of 10 with a chlorobenzoxazole to provide 3 (MK-4305), a potent dual orexin receptor antagonist that is currently being tested in phase III clinical trials for the treatment of primary insomnia.[1] Orexin receptor antagonism represents a novel approach for the treatment of insomnia that directly targets sleep/wake regulation. Several such compounds have entered into clinical development, including the dual orexin receptor antagonists, suvorexant and almorexant. In this study, we have used equilibrium and kinetic binding studies with the orexin-2 (OX₂) selective antagonist radioligand, [³H]-EMPA, to profile several orexin receptor antagonists. Furthermore, selected compounds were studied in cell-based assays of inositol phosphate accumulation and ERK-1/2 phosphorylation in CHO cells stably expressing the OX2 receptor that employ different agonist incubation times (30 and 5 min, respectively). EMPA, suvorexant, almorexant and TCS-OX-29 all bind to the OX₂ receptor with moderate to high affinity (pk(I) values ≥ 7.5), whereas the primarily OX1 selective antagonists SB-334867 and SB-408124 displayed low affinity (pK(I) values ca. 6). Competition kinetic analysis showed that the compounds displayed a range of dissociation rates from very fast (TCS-OX2-29, k(off) = 0.22 min⁻¹) to very slow (almorexant, k(off) = 0.005 min⁻¹). Notably, there was a clear correlation between association rate and affinity. In the cell-based assays, fast-offset antagonists EMPA and TCS-OX2-29 displayed surmountable antagonism of orexin-A agonist activity. However, both suvorexant and particularly almorexant cause concentration-dependent depression in the maximal orexin-A response, a profile that is more evident with a shorter agonist incubation time. Analysis according to a hemi-equilibrium model suggests that antagonist dissociation is slower in a cellular system than in membrane binding; under these conditions, almorexant effectively acts as a pseudo-irreversible antagonist.[2]

Solubility Data

Solubility (In Vitro)	DMSO: ~10 mg/mL (~22.2 mM) Water: <1 mg/mL Ethanol: <1 mg/mL
Solubility (In Vivo)	4% DMSO+10% PEG 400+10% Tween 80: 5mg/mL  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.2177 mL	11.0884 mL	22.1769 mL
	5 mM	0.4435 mL	2.2177 mL	4.4354 mL
	10 mM	0.2218 mL	1.1088 mL	2.2177 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.