| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
Skeletal muscle myosin-2 (also known as non-muscle myosin heavy chain IIA or skeletal muscle myosin II). (R)-MPH-220 is a selective inhibitor of skeletal muscle myosin-2 ATPase activity.
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| ln Vitro |
(R)-MPH-220 is a selective inhibitor of skeletal muscle myosin-2. By inhibiting myosin-2 ATPase activity, the compound prevents the binding of myosin to actin, thereby reducing the formation of cross-bridges and leading to muscle relaxation. The specific IC₅0 values for myosin-2 inhibition are not provided in the available data. (R)-MPH-220 is expected to have similar or greater potency compared to the racemic MPH-220 mixture.
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| Enzyme Assay |
A general cell-free protocol for assessing skeletal muscle myosin-2 ATPase activity: An NADH-coupled ATPase assay is used. Skeletal muscle myosin-2 is purified from rabbit or rat skeletal muscle or obtained as a recombinant protein. The myosin ATPase activity is measured in a buffer containing 20 mM MOPS (pH 7.0), 5 mM MgCl2, 0.5 mM DTT, 0.3 mM NADH, 2 mM phosphoenolpyruvate (PEP), 20 U/mL lactate dehydrogenase, 20 U/mL pyruvate kinase, and various concentrations of (R)-MPH-220 (0.01 nM to 100 uM). The reaction is initiated by adding 1 mM ATP. The decrease in absorbance at 340 nm (due to oxidation of NADH to NAD+) is monitored for 10 minutes at 25degC. The rate of NADH oxidation is proportional to the ATPase activity. The IC₅0 is calculated from the inhibition curve. For selectivity profiling, the compound is tested against other myosin isoforms (e.g., cardiac myosin-2, smooth muscle myosin-2, non-muscle myosin-2A/B).
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| Cell Assay |
A general cellular protocol for assessing muscle relaxation: Skeletal muscle cells (e.g., C2C12 mouse myoblasts) are differentiated into myotubes in DMEM containing 2% horse serum for 5-7 days. The differentiated myotubes are treated with various concentrations of (R)-MPH-220 (0.1, 1, 10, 50, 100 uM) for 24 hours. The relaxation of the myotubes is assessed by measuring the contractile force using a cell contractility assay (e.g., a collagen gel contraction assay). The myotubes are embedded in a collagen gel, and the diameter of the gel is measured after 24 hours of treatment. For functional assays, electrically stimulated muscle contraction is measured using a muscle organ bath. The compound is added to the organ bath, and the twitch force is recorded. The EC₅0 for relaxation is calculated. Calcium imaging can also be performed using a calcium-sensitive dye (e.g., Fura-2 AM) to assess whether the compound affects calcium signaling.
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| Animal Protocol |
A general animal protocol for evaluating the antispasmodic effects of (R)-MPH-220 in a mouse model of muscle spasticity: A mouse model of muscle spasticity is induced by intramuscular injection of 2% carrageenan into the gastrocnemius muscle, or by the use of a mouse model of spinal cord injury (SCI) or cerebral palsy. Male CD-1 mice (25-30 g) are used. (R)-MPH-220 is formulated in a suitable vehicle (e.g., 0.5% methylcellulose or 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline) and administered via oral gavage at doses of 1, 3, 10, and 30 mg/kg once daily for 7 days. A vehicle control group and a positive control group (e.g., baclofen, 10 mg/kg) are included. Muscle spasticity is assessed by measuring muscle resistance to passive stretch using a force gauge (modified Ashworth scale). Motor coordination is assessed using the rotarod test. Grip strength is measured using a grip strength meter. At the end of the study, the mice are euthanized, and the gastrocnemius muscle is excised and weighed. A section of the muscle is processed for histopathological examination (H&E staining). The muscle is also homogenized, and levels of inflammation markers (TNF-alpha, IL-1beta, IL-6) are measured by ELISA.
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| ADME/Pharmacokinetics |
General pharmacokinetic protocol for (R)-MPH-220: Male Sprague-Dawley rats are administered (R)-MPH-220 via oral gavage (PO, 10 mg/kg) and intravenous (IV, 2 mg/kg) injection. The compound is formulated in a vehicle such as 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline. Blood samples are collected at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, 12, and 24 hours post-dose. Plasma concentrations of (R)-MPH-220 are quantified by LC-MS/MS. PK parameters (Cmax, Tmax, AUC, t½, clearance, Vd, and oral bioavailability) are calculated using non-compartmental analysis. MPH-220 is reported to be orally active, so (R)-MPH-220 is expected to have good oral bioavailability.
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| Toxicity/Toxicokinetics |
General toxicity protocol for (R)-MPH-220: A 14-day repeated-dose oral toxicity study is performed in Sprague-Dawley rats. (R)-MPH-220 is administered via oral gavage at doses of 10, 30, and 100 mg/kg/day for 14 consecutive days. Clinical signs, body weight, and food consumption are monitored daily. At the end of the study, blood samples are collected for hematology (complete blood count with differential) and serum chemistry (ALT, AST, ALP, BUN, creatinine, total protein, albumin, glucose, CPK). Gross necropsy is performed, and the weights of major organs (liver, kidney, spleen, heart, lung, brain, skeletal muscle (gastrocnemius)) are recorded. Histopathological examination of these organs is conducted, with special attention to skeletal muscle for any signs of myopathy or degeneration.
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| References | |
| Additional Infomation |
(R)-MPH-220 is the R-isomer of MPH-220. MPH-220 is a selective, orally active inhibitor of skeletal muscle myosin-2. (R)-MPH-220 is a chiral compound with a specific (4aR)-configuration. It relaxes skeletal muscle and acts as an antispasmodic agent, making it useful for research into spasms and muscle stiffness. The compound appears as an orange to red solid with a purity of 99.09%. The molecular structure is not provided in the available data.
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| Molecular Formula |
C20H21N3O3S
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| Molecular Weight |
383.46
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| Appearance |
Solid powder
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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) |
DMSO : 100 mg/mL (260.78 mM; with sonication and heat)
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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 | 2.6078 mL | 13.0392 mL | 26.0783 mL | |
| 5 mM | 0.5216 mL | 2.6078 mL | 5.2157 mL | |
| 10 mM | 0.2608 mL | 1.3039 mL | 2.6078 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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