| Size | Price | Stock | Qty |
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| 100mg |
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| Targets |
Ramelteon metabolite M-II targets the human melatonin receptors MT1 and MT2, which are G protein-coupled receptors involved in the regulation of circadian rhythms and sleep-wake cycles. It acts as an agonist at these receptors. The compound exhibits IC50 values of 208 pM for MT1 and 1470 pM for MT2. Its affinity for MT1 receptors is 10-fold lower than ramelteon and 2.5-fold lower than melatonin; for MT2 receptors, it is approximately 5-fold and 1.5-fold lower, respectively. The compound shows no significant affinity for 215 other targets, except for the 5-HT2B receptor (Ki = 1.75+/-0.23 microM).
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| ln Vitro |
The affinity of Ramelteon metabolite M-II (M-II) for the MT1 receptor is 2.5 times lower than that of melatonin, and it is ten times lower than that of Ramelteon. Similarly, M-II has a 1.5-fold lower affinity for the MT2 receptor than melatonin, and a 5-fold lower affinity for ramelteon. Even at doses up to 10 μM, the ramelteon metabolite M-II exhibits little affinity for quinone reductase 2. Moreover, research was done on the selectivity of Ramelteon metabolite M-II for melatonin receptors in comparison to 215 targets, which included enzymes, transporters, ion channels, and other receptors. With a Ki value of 1.75±0.23 μM, ramelteon metabolite M-II exhibits no appreciable affinity or activity towards other targets, with the exception of the 5-HT2B receptor. The metabolite M-II of Ramelteon is around 17- and 4.3-fold less active than melatonin at MT1 receptors, respectively. Similarly, compared to melatonin and Ramelteon, the metabolite M-II of Ramelteon is around 1.6 and 28 times less effective at MT2 receptors [1].
In vitro studies have characterized Ramelteon metabolite M-II as a potent melatonin receptor agonist. Its affinity for MT1 receptors is approximately 17-fold lower than ramelteon and 4.3-fold lower than melatonin, while for MT2 receptors, it is approximately 28-fold and 1.6-fold lower, respectively. The compound has been evaluated in binding assays against a panel of 215 targets, including other receptors, transporters, ion channels, and enzymes, and showed no significant affinities or activities except for the 5-HT2B receptor. Its selectivity profile supports its role as a specific melatonin receptor agonist. |
| ln Vivo |
NREM sleep was greatly increased (F1,7=96.3, p<0.01) and wakefulness was significantly reduced (F1,7=56.7, p<0.01) when ramelteon metabolite M-II (1 mg/kg) was administered. Additionally, similar effects were obtained with smaller doses of M-II (0.1 mg/kg) (F1,7=121.9, p<0.01 for NREM; F1,7=87.0, p<0.01), with decreased wakefulness lasting for 6 hours following administration of either dose. Following the delivery of 0.01 mg/kg Ramelteon metabolite M-II, only NREM sleep showed a significant increase (F1,7=10.5, p<0.05). After administering any dose of M-II evaluated in this investigation, no discernible changes were seen in REM sleep [1].
In vivo, Ramelteon metabolite M-II has been shown to significantly increase non-rapid eye movement (NREM) sleep and decrease wakefulness in animal models. In a rat study, administration of M-II at 1 mg/kg significantly increased NREM sleep and significantly decreased wakefulness. A lower dose of 0.1 mg/kg produced similar results, with decreased wakefulness sustained for 6 hours after administration. At 0.01 mg/kg, only NREM sleep was significantly increased. No significant differences in REM sleep were observed at any of the doses tested, confirming its role as a sleep-promoting metabolite. |
| Enzyme Assay |
The in vitro receptor binding assay for Ramelteon metabolite M-II typically involves competitive binding studies using membrane preparations from cells expressing human MT1 or MT2 melatonin receptors. Radiolabeled 2-[125I]-iodomelatonin is used as the tracer ligand. Varying concentrations of M-II are incubated with the membrane preparation and the radioligand. Bound radioactivity is measured after filtration or centrifugation. IC50 values are determined from competition curves and converted to Ki values. The assay is performed in triplicate with appropriate controls to ensure data accuracy.
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| Cell Assay |
In vitro cellular assays for Ramelteon metabolite M-II are conducted using cells expressing human MT1 or MT2 receptors, such as CHO or HEK293 cells. Cells are treated with varying concentrations of M-II, and receptor activation is assessed by measuring the inhibition of forskolin-stimulated cAMP accumulation, a downstream effect of melatonin receptor activation. The EC50 values are determined from dose-response curves. The potency of M-II is compared to that of ramelteon and melatonin to assess relative efficacy. Selectivity is confirmed by testing against a panel of other receptors and enzymes.
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| Animal Protocol |
In vivo animal studies for Ramelteon metabolite M-II are typically performed in rodents, such as rats, to evaluate its effects on sleep-wake architecture. Animals are surgically implanted with electrodes for electroencephalography (EEG) and electromyography (EMG) recording. M-II is administered via intraperitoneal or oral routes at various doses (e.g., 0.01, 0.1, and 1 mg/kg). Sleep-wake states are scored based on EEG/EMG signals, and the duration of NREM sleep, REM sleep, and wakefulness is quantified. The compound's effects on sleep parameters are compared to vehicle control and ramelteon.
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| ADME/Pharmacokinetics |
Ramelteon metabolite M-II has a molecular weight of 275.34 g/mol and a molecular formula of C16H21NO3. As the primary metabolite of ramelteon, it is formed through hydroxylation and is the major circulating species in serum. The compound is a racemic mixture of R and S isomers at the hydroxy position. Its pharmacokinetic profile is characterized by rapid formation from the parent drug and a relatively short half-life. It is typically used as a reference standard in analytical method development for ramelteon and its metabolites.
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| Toxicity/Toxicokinetics |
Toxicology data for Ramelteon metabolite M-II are derived from studies of the parent drug ramelteon. As a melatonin receptor agonist, the compound's safety profile is consistent with its mechanism of action. Preclinical toxicology studies have evaluated the effects of ramelteon and its metabolites on reproductive function, neurobehavioral parameters, and general toxicity. No significant toxicity has been attributed specifically to M-II. The compound is generally well-tolerated at therapeutic concentrations. Standard toxicological assessments have been conducted to support the clinical use of ramelteon.
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| References | |
| Additional Infomation |
Ramelteon metabolite M-II is the primary active metabolite of the insomnia drug ramelteon. It is a potent and selective agonist at melatonin MT1 and MT2 receptors, with IC50 values of 208 pM and 1470 pM, respectively. M-II significantly increases NREM sleep and decreases wakefulness in animal models. It is a racemic mixture of R and S isomers at the hydroxy position and serves as a reference standard for pharmacokinetic studies and analytical method development. The compound is for research use only.
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| Molecular Formula |
C16H21NO3
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| Molecular Weight |
275.34284
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| Exact Mass |
275.152
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| CAS # |
896736-21-3
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| Related CAS # |
Ramelteon metabolite M-II-d3;1246812-22-5
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| PubChem CID |
46783414
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| Appearance |
White to off-white solid powder
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| Density |
1.199g/cm3
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| Boiling Point |
495.171ºC at 760 mmHg
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| Melting Point |
96-98ºC
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| Flash Point |
253.271ºC
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| Index of Refraction |
1.576
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| LogP |
1.929
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
20
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| Complexity |
360
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| Defined Atom Stereocenter Count |
1
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| SMILES |
CC(C(=O)NCC[C@@H]1CCC2=C1C3=C(C=C2)OCC3)O
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| InChi Key |
FGFNIJYHXMJYJN-KFJBMODSSA-N
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| InChi Code |
InChI=1S/C16H21NO3/c1-10(18)16(19)17-8-6-12-3-2-11-4-5-14-13(15(11)12)7-9-20-14/h4-5,10,12,18H,2-3,6-9H2,1H3,(H,17,19)/t10?,12-/m0/s1
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| Chemical Name |
2-hydroxy-N-[2-[(8S)-2,6,7,8-tetrahydro-1H-cyclopenta[e][1]benzofuran-8-yl]ethyl]propanamide
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| HS Tariff Code |
2934.99.9001
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| 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)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.6319 mL | 18.1594 mL | 36.3187 mL | |
| 5 mM | 0.7264 mL | 3.6319 mL | 7.2637 mL | |
| 10 mM | 0.3632 mL | 1.8159 mL | 3.6319 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.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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