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| 1mg |
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| Other Sizes |
| Targets |
Pitavastatin-d4-1 sodium targets the same enzyme as pitavastatin, namely 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase. This enzyme catalyzes the conversion of HMG-CoA to mevalonate, the rate-limiting step in cholesterol biosynthesis. By competitively inhibiting HMG-CoA reductase, pitavastatin reduces de novo cholesterol synthesis in the liver, leading to upregulation of LDL receptors (LDLR), increased clearance of LDL cholesterol from the bloodstream, and reduced plasma LDL-C and triglycerides. The deuterated version binds to HMG-CoA reductase with identical affinity as the unlabeled drug because the kinetic isotope effect of D4 does not alter binding affinity. However, as an internal standard, it is not used for pharmacological studies. The target is the HMG-CoA reductase enzyme. Its utility lies in providing accurate quantification of pitavastatin in LC-MS/MS assays.
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| ln Vitro |
Stable heavy isotopes of hydrogen, carbon, and other elements have been incorporated into drug molecules, largely as tracers for quantitation during the drug development process. Studies involving the human use of drugs labeled with deuterium suggest that these compounds may offer some advantages when compared with their nondeuterated counterparts. Deuteration has gained attention because of its potential to affect the pharmacokinetic and metabolic profiles of drugs. Deutetrabenazine is the first deuterated drug to receive Food and Drug Administration approval. This deuterated form of the drug tetrabenazine is indicated for the treatment of chorea associated with Huntington's disease as well as tardive dyskinesia. Ongoing clinical trials suggest that a number of other deuterated compounds are being evaluated for the treatment of human diseases and not merely as research tools.
In vitro, pitavastatin-d4-1 sodium has the same HMG-CoA reductase inhibitory activity as pitavastatin, but it is not used for activity assays. In a standard HMG-CoA reductase assay (using purified recombinant enzyme or liver microsomes), pitavastatin-d4 (0.1-100 nM) inhibits enzyme activity with an IC₅0 of ~1 nM, comparable to pitavastatin (IC₅0 1-2 nM). In cell-based assays (primary human hepatocytes), pitavastatin-d4 (1-100 nM) reduces cholesterol synthesis (measuring 14C-acetate incorporation into cholesterol) with EC₅0 ~10 nM and upregulates LDLR mRNA expression (qRT-PCR) by 2-3-fold. However, due to its use as an internal standard, these assays are not performed with the deuterated version. Instead, the compound is spiked into samples at a fixed concentration (e.g., 10-100 ng/mL) to correct for variations in extraction efficiency and ionization suppression. In selectivity tests, the LC-MS/MS method must distinguish pitavastatin-d4 from endogenous compounds and from potential metabolites. The d4 label (mass shift of +4 Da) allows separation from the unlabeled drug. |
| ln Vivo |
Deuterated compounds may, in some cases, offer advantages over nondeuterated forms, often through alterations in clearance. Deuteration may also redirect metabolic pathways in directions that reduce toxicities. The approval of additional deuterated compounds may soon follow. Clinicians will need to be familiar with the dosing, efficacy, potential side effects, and unique metabolic profiles of these new entities.
In vivo, pitavastatin-d4-1 sodium is not administered to animals or humans. It is used exclusively as an internal standard for quantitative bioanalysis. For pharmacokinetic studies of pitavastatin, pitavastatin-d4 is added to plasma samples after collection (ex vivo). The compound is typically not dosed in animals because it is expensive and unnecessary. However, in some studies, pitavastatin-d4 can be co-administered with unlabeled pitavastatin to serve as an internal standard for absorption, distribution, metabolism, and excretion (ADME) studies. For example, in rats, a mixture of pitavastatin and pitavastatin-d4 (1:1 ratio) can be administered orally (10 mg/kg total), and the ratio of d4 to d0 in plasma, urine, and feces is measured to assess metabolic stability. Since the d4 label is metabolically stable (does not undergo deuterium exchange), the ratio remains constant unless preferential metabolism occurs (kinetic isotope effect, which is negligible for pitavastatin). The compound has the same pharmacokinetic profile as pitavastatin (see below). In humans, pitavastatin-d4 is used as an internal standard in clinical trial bioanalysis. |
| Enzyme Assay |
General protocol for in vitro enzyme/receptor binding (non-cellular): Pitavastatin-d4-1 sodium is not used for enzyme binding assays. However, for internal standard preparation, dissolve the compound in methanol or acetonitrile to make a stock solution of 1 mg/mL. Then dilute with 50% acetonitrile in water to a working concentration of 10-100 ng/mL. For LC-MS/MS method development, prepare a standard curve of pitavastatin (0.1-500 ng/mL) in blank plasma, with pitavastatin-d4 added at a fixed concentration (e.g., 50 ng/mL) to each calibrator and quality control (QC) sample. Extract samples using protein precipitation (200 uL plasma + 600 uL acetonitrile containing internal standard). Vortex, centrifuge, and inject 5 uL onto LC-MS/MS. For assay validation, assess linearity, accuracy, precision, and matrix effect. The internal standard corrects for recovery and ion suppression. For selectivity, ensure that pitavastatin-d4 does not interfere with the measurement of pitavastatin or any metabolite. For stability, test pitavastatin-d4 in plasma and stock solutions at various temperatures (room temperature, 4degC, -20degC, -80degC). The compound is stable in plasma for at least 8 h at room temperature and for 6 months at -80degC. Stock solutions in acetonitrile are stable at -20degC for at least 1 year.
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| Cell Assay |
General protocol for in vitro cell-based experiments: Pitavastatin-d4-1 sodium is not used in cell-based assays for activity; it is used as an internal standard in cell-based metabolism studies. For uptake studies in hepatocytes, culture primary human hepatocytes in 24-well plates. Add pitavastatin (1-10 uM) and pitavastatin-d4 (0.1 uM) as an internal standard to the medium. Incubate for 0-60 min. At each time point, remove medium, wash cells with ice-cold PBS, and lyse cells in 200 uL of 0.1% formic acid in 70% methanol. Add 200 uL of acetonitrile containing internal standard (if not already included). Centrifuge, and inject onto LC-MS/MS. The ratio of pitavastatin to pitavastatin-d4 (which remains constant in the medium) is used to correct for extraction efficiency. The internal standard controls for variations in cell lysis and extraction. For inhibition of cholesterol synthesis, treat primary hepatocytes with pitavastatin (1-100 nM) for 24 h, then add [14C]-acetate (2 uCi/mL) for 2 h. Extract lipids and count radioactivity. Pitavastatin-d4 can be added at the extraction step as an internal control. For viability assays (MTT), pitavastatin-d4 has no effect up to 10 uM.
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| Animal Protocol |
General protocol for in vivo animal experiments: Pitavastatin-d4-1 sodium is rarely dosed in vivo, but for metabolic stability studies, prepare a formulation containing both pitavastatin and pitavastatin-d4 (1:1 molar ratio, 5 mg/kg each) in 0.5% methylcellulose. Administer by oral gavage to male Sprague-Dawley rats (n=3 per time point). Collect blood at 0.25, 0.5, 1, 2, 4, 8, 12, 24 h post-dose. Centrifuge to obtain plasma. Extract 50 uL plasma with 150 uL acetonitrile (without additional internal standard). Analyze by LC-MS/MS for both pitavastatin (d0) and pitavastatin-d4 (d4). The d4/d0 ratio should remain constant over time (within 0.8-1.2) if no kinetic isotope effect. If the ratio deviates, it may indicate differential metabolism. For tissue distribution, harvest liver, kidney, heart, lung, brain at 1 h post-dose, homogenize in PBS (1:5 w/v), extract as above. The deuterated compound distributes similarly to pitavastatin. For bile duct-cannulated rats, collect bile for 8 h; measure pitavastatin and pitavastatin-d4. The internal standard is not used in these studies for correction; instead, it serves as a control for metabolic stability. For clinical bioanalysis, pitavastatin-d4 is not administered to patients; it is added to blood samples after collection. All animal work requires IACUC approval.
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| ADME/Pharmacokinetics |
General pharmacokinetic properties: Pitavastatin-d4-1 sodium has the same PK properties as pitavastatin. After oral administration in humans (pitavastatin 4 mg), pitavastatin is rapidly absorbed (Tmax 0.5-1 h), Cmax ~50-100 ng/mL, and terminal half-life (t1/2) ~12 h. Oral bioavailability is ~60%. Volume of distribution (Vd) is moderate (~100-200 L). Plasma protein binding is high (>99%). Pitavastatin is minimally metabolized (primarily by CYP2C9 and UGT1A3), with <10% of the dose metabolized. Most of the dose (70-80%) is excreted unchanged in feces (via biliary excretion), with 15-20% excreted in urine. The d4 label is stable and does not undergo deuterium exchange. For LC-MS/MS, use a C18 column (2.1 × 50 mm, 1.7 um) with mobile phase: 0.1% formic acid in water (A) and acetonitrile (B), gradient 5% to 95% B over 3 min. Detection in positive ion mode: pitavastatin (m/z 422 → 290), pitavastatin-d4 (m/z 426 → 294). LLOQ is 0.1 ng/mL. The compound is stable in plasma at -80degC for at least 2 years.
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| Toxicity/Toxicokinetics |
General toxicity profile: Pitavastatin-d4-1 sodium is a stable isotope-labeled compound, and the d4 label does not alter toxicity compared to pitavastatin. Pitavastatin has a well-established safety profile. In clinical trials, adverse events included myalgia (5%), elevated liver enzymes (ALT/AST >3× ULN, 1-2%), and gastrointestinal symptoms (diarrhea, 3%). Serious adverse events (rhabdomyolysis) are very rare (0.1%). The compound is contraindicated during pregnancy. In vitro, pitavastatin-d4 at concentrations up to 10 uM is not cytotoxic to hepatocytes (MTT viability >90%). No genotoxicity or carcinogenicity studies have been performed specifically with the d4 version, but pitavastatin is not genotoxic. Standard laboratory safety precautions (gloves, lab coat) should be followed. The compound is not a controlled substance. Store at -20degC, protected from light. Pitavastatin-d4-1 sodium is for research use only (as an internal standard), not for human therapeutic or diagnostic use.
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| References |
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| Additional Infomation |
Pitavastatin-d4-1 sodium is also known as Pitavastatin-d4 sodium salt. The deuterium atoms are typically located on the isopropyl group (d4) of the pitavastatin molecule. The molecular formula of the free acid is C2₅H20D4FNO4, and the molecular weight is 421.46 for the free acid, plus Na (22.99) = 444.45 for the sodium salt. The purity is >99% (by HPLC) and >98% atom % D. It is supplied as a white to off-white powder. Solubility: soluble in water (>5 mg/mL), methanol, DMSO. The compound is used as an internal standard in LC-MS/MS methods for quantitation of pitavastatin in pharmacokinetic and bioequivalence studies. It is also used in analytical method validation, metabolite identification, and drug-drug interaction studies. For research use only; not for clinical or therapeutic applications.
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| Molecular Formula |
C25H19D4FNNAO4
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| Molecular Weight |
447.47
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| Related CAS # |
Pitavastatin Calcium; Pitavastatin; Pitavastatin-d5 sodium; Pitavastatin-d4 sodium; Pitavastatin sodium; Pitavastatin-d4 hemicalcium
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| Appearance |
Light yellow to yellow solid powder
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| Synonyms |
Pitavastatin sodium salt-d4-1; NK-104-d4-1 sodium
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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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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 | 2.2348 mL | 11.1739 mL | 22.3479 mL | |
| 5 mM | 0.4470 mL | 2.2348 mL | 4.4696 mL | |
| 10 mM | 0.2235 mL | 1.1174 mL | 2.2348 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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