| Size | Price | Stock | Qty |
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| 5mg |
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| Other Sizes |
| Targets |
(S)-4-Hydroxymephenytoin has no therapeutic target itself. It is a pharmacogenomic probe for CYP2C19, a critical drug-metabolizing enzyme. CYP2C19 is responsible for the metabolism of many clinically used drugs, including proton pump inhibitors (e.g., omeprazole), antidepressants (e.g., citalopram), antiplatelet drugs (e.g., clopidogrel), and antifungals (e.g., voriconazole). The rate of formation of (S)-4-Hydroxymephenytoin directly reflects the enzymatic activity and genetic polymorphisms of CYP2C19, which classifies individuals as extensive, intermediate, or poor metabolizers.
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| ln Vitro |
The compound is not pharmacologically active and has no direct enzyme inhibition activity. It is used as a surrogate marker for CYP2C19 activity. In pooled human liver microsomes, the formation clearance (CLint) of (S)-4-Hydroxymephenytoin is highly correlated (R2 > 0.9) with the rate of omeprazole 5-hydroxylation. It exhibits no cytotoxicity in primary human hepatocytes. It is stable in plasma and urine and does not induce CYP450 enzymes. It is used as a reference standard for activity-based protein profiling.
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| ln Vivo |
In vivo, (S)-4-Hydroxymephenytoin is a clinical probe drug for phenotyping CYP2C19. To determine a patient's CYP2C19 phenotype, a single oral dose of racemic mephenytoin (100 mg) is administered. Urine is collected for 0-8 hours. The ratio of (S)-mephenytoin to (S)-4-Hydroxymephenytoin in the urine (hydroxylation index) is calculated by GC or HPLC. A high ratio indicates a poor metabolizer phenotype (CYP2C19 deficiency). It is also used in drug-drug interaction studies to evaluate if a new drug candidate inhibits CYP2C19 activity in vivo.
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| Enzyme Assay |
CYP2C19 enzyme assay is performed using pooled human liver microsomes (0.2 mg protein/mL) or recombinant human CYP2C19. The incubation mixture contains 0.1 M Tris-HCl buffer (pH 7.4), 10 mM MgCl2, 1 mM NADPH, and varying concentrations of (S)-mephenytoin (1-1000 uM) as a substrate. The reaction is initiated by adding NADPH, incubated at 37degC for 30 min, and terminated with acetonitrile. The formation of (S)-4-Hydroxymephenytoin is quantified by HPLC-UV (210 nm) or LC-MS/MS (MRM transition 235→150). The kinetic parameters (Km, Vmax) are calculated.
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| Cell Assay |
In vitro CYP2C19 inhibition screening can be performed in human liver microsomes. The test compound is pre-incubated with microsomes and NADPH for 15 min. Then, (S)-mephenytoin (50 uM, ~Km concentration) is added. After a 30-min incubation, the formation of (S)-4-Hydroxymephenytoin is measured. The % inhibition is calculated relative to a control (DMSO). Ticlopidine (1 uM) is used as a positive control for CYP2C19 inhibition. The IC₅0 is calculated from a dose-response curve. The compound is stable in microsomal buffer.
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| Animal Protocol |
In vivo CYP2C19 phenotyping is performed in male Sprague-Dawley rats or human volunteers. A single oral dose of mephenytoin (10 mg/kg for rats, 100 mg for humans) is administered. Urine is collected for 8-24 h. The concentration of (S)-4-Hydroxymephenytoin in urine is measured by GC-MS or LC-MS. The molar ratio of (S)-mephenytoin to (S)-4-Hydroxymephenytoin is calculated. For drug-drug interaction studies, the test compound is co-administered with mephenytoin. A significant increase in the ratio (> 2-fold) compared to the control indicates that the test compound is an inhibitor of CYP2C19 in vivo.
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| ADME/Pharmacokinetics |
(S)-4-Hydroxymephenytoin has a molecular weight of 234.25 and is a polar, hydrophilic molecule. It is not intended for direct therapeutic administration. In pharmacokinetic studies where mephenytoin is administered, (S)-4-Hydroxymephenytoin is rapidly formed in the liver and excreted in the urine. The elimination half-life of the metabolite in plasma is short (approx. 2-4 h). It is primarily cleared by the kidneys via glomerular filtration. It does not accumulate in tissues. It is not metabolized further to a significant extent. The compound is an analytical standard.
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| Toxicity/Toxicokinetics |
(S)-4-Hydroxymephenytoin has low intrinsic toxicity. It is a normal metabolite of mephenytoin, which has been used clinically as an anticonvulsant. At the doses used for phenotyping (trace amounts), it does not cause adverse effects. However, the parent drug mephenytoin may cause dose-related neurological effects. As a laboratory chemical, standard precautions apply: wear PPE (gloves, lab coat, eye protection), avoid inhalation and skin contact. (S)-4-Hydroxymephenytoin is not classified as a hazardous waste.
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| References | |
| Additional Infomation |
(S)-4-Hydroxymephenytoin ((S)-4′-Hydroxy Mephenytoin; CAS# 82695-93-0) is a research-grade pharmacogenomic standard. It is not an FDA-approved drug. It is the key biomarker used for phenotyping the drug-metabolizing enzyme CYP2C19 to guide personalized medicine, particularly for drugs like clopidogrel and proton pump inhibitors. For research use only, not for diagnostic or therapeutic applications. Storage: powder at -20degC, protected from light.
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| Molecular Formula |
C12H14N2O3
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| Molecular Weight |
234.25
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| Exact Mass |
234.1
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| CAS # |
82695-93-0
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| Related CAS # |
4-Hydroxymephenytoin; 61837-65-8
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| PubChem CID |
40572060
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| Appearance |
Typically exists as solids at room temperature
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| Hydrogen Bond Donor Count |
2
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
17
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| Complexity |
337
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| Defined Atom Stereocenter Count |
1
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| SMILES |
CC[C@@]1(C(=O)N(C(=O)N1)C)C2=CC=C(C=C2)O
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| InChi Key |
OQPLORUDZLXXPD-LBPRGKRZSA-N
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| InChi Code |
InChI=1S/C12H14N2O3/c1-3-12(8-4-6-9(15)7-5-8)10(16)14(2)11(17)13-12/h4-7,15H,3H2,1-2H3,(H,13,17)/t12-/m0/s1
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| Chemical Name |
(5S)-5-ethyl-5-(4-hydroxyphenyl)-3-methylimidazolidine-2,4-dione
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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 | 4.2689 mL | 21.3447 mL | 42.6894 mL | |
| 5 mM | 0.8538 mL | 4.2689 mL | 8.5379 mL | |
| 10 mM | 0.4269 mL | 2.1345 mL | 4.2689 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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