| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
Amlodipine-1,1,2,2-d4 maleate is a stable isotope-labeled internal standard that binds to the same target as its non-labeled parent, Amlodipine maleate. Amlodipine is a dihydropyridine calcium channel blocker (CCB) that acts by binding to and blocking the voltage-dependent L-type calcium channels (Cav1.2). These channels are located on the smooth muscle cells of blood vessels and cardiac muscle. By inhibiting the initial influx of calcium ions (Ca2+) through these channels, Amlodipine causes vasodilation of the peripheral arteries (arterioles) and, to a lesser extent, coronary arteries. This vasodilation reduces peripheral vascular resistance (afterload), thereby lowering systemic blood pressure. It also reduces myocardial oxygen demand, which is beneficial for treating angina. The labeled version mimics this binding but is used for analytical quantification, not for pharmacological effect.
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| ln Vitro |
Drug compounds have included stable heavy isotopes of carbon, hydrogen, and other elements, mostly as quantitative tracers while the drugs were being developed. Because deuteration may have an effect on a drug's pharmacokinetics and metabolic properties, it is a cause for concern [1].
Amlodipine-1,1,2,2-d4 maleate itself is not used for in vitro bioassays, but its unlabeled counterpart, Amlodipine, exhibits well-defined activity. In vitro, Amlodipine potently inhibits the contraction of rat aortic rings induced by potassium chloride (KCl) or norepinephrine, with an IC₅0 in the low nanomolar range (approx. 1-10 nM). In cultured vascular smooth muscle cells (VSMCs), Amlodipine (0.1-10 uM) blocks the influx of calcium stimulated by vasoconstrictors, leading to reduced cell proliferation and migration. Additionally, Amlodipine has been shown to increase nitric oxide (NO) bioavailability in endothelial cells, contributing to its vasodilatory effects. It also exhibits antioxidant properties by reducing the production of reactive oxygen species (ROS) in these cell lines. The labeled Amlodipine-d4 serves as an internal standard in LC-MS studies to accurately measure the drug concentration in the culture media to which the cells are exposed. |
| ln Vivo |
Amlodipine-1,1,2,2-d4 maleate is a stable isotope-labeled internal standard, and its in vivo activity is the same as its non-labeled parent compound. In animal models of hypertension, such as spontaneously hypertensive rats (SHRs), oral administration of Amlodipine maleate (1-10 mg/kg/day) produces a dose-dependent and long-lasting reduction in systolic and diastolic blood pressure, lasting over 24 hours. In dog models of angina, intravenous Amlodipine (0.1-1 mg/kg) increases coronary blood flow and reduces myocardial oxygen consumption. Due to its slow onset of action and long duration, it does not cause reflex tachycardia. The labeled Amlodipine-1,1,2,2-d4 maleate is used as an internal standard in these animal studies to accurately quantify the plasma and tissue levels of the drug by LC-MS, allowing for precise correlation of the drug concentration with the observed antihypertensive effects (PK/PD modeling).
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| Enzyme Assay |
A generic non-cell-based assay for Amlodipine-1,1,2,2-d4 maleate is its use as an internal standard in an LC-MS/MS method. Prepare a standard stock solution of the unlabeled Amlodipine maleate in methanol (1 mg/mL). Prepare a separate stock solution of the internal standard (Amlodipine-1,1,2,2-d4 maleate) at the same concentration. Prepare calibration standards by spiking the unlabeled analyte into a blank matrix (e.g., blank human plasma) to achieve concentrations ranging from 0.1 ng/mL to 100 ng/mL. Add a fixed concentration of the internal standard (e.g., 10 ng/mL) to each calibration standard. Also prepare blank and double-blank samples. For sample preparation, perform liquid-liquid extraction: add 50 uL of the sample to 150 uL of tert-butyl methyl ether, vortex, and centrifuge. Transfer the organic layer (top) to a clean tube and evaporate under nitrogen. Reconstitute the residue in 150 uL of mobile phase. Analyze by LC-MS/MS in positive ion mode. Monitor the mass transitions: m/z 409.2 → 238.1 for Amlodipine, and m/z 413.2 → 242.1 for Amlodipine-d4. Construct the calibration curve by plotting the peak area ratio (analyte/IS) vs. the nominal concentration.
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| Cell Assay |
A standard in vitro cell-based assay for the unlabeled Amlodipine involves measuring its effects on calcium influx in vascular smooth muscle cells (VSMCs). Isolate rat aortic VSMCs and culture them in DMEM with 10% FBS. Seed the cells in 96-well plates at 2×10⁴ cells/well and allow them to become confluent. Load the cells with a fluorescent calcium indicator, such as Fluo-4 AM, in Hanks‘ balanced salt solution (HBSS) for 30 minutes at 37degC. Wash the cells to remove excess dye. Measure the fluorescence (Ex/Em = 494/506 nm) using a fluorescence plate reader. Add various concentrations of unlabeled Amlodipine (0.1 nM to 10 uM) to the cells and measure baseline fluorescence for 2 minutes. Then, add a vasoconstrictor (e.g., 50 mM KCl or 1 uM angiotensin II) to stimulate calcium influx. Record the change in fluorescence intensity. Calculate the percentage inhibition of the calcium peak signal relative to the control. Determine the half-maximal inhibitory concentration (IC₅0) from the dose-response curve. Use Amlodipine-d4 as an internal standard to quantify the actual drug concentration in the HBSS by LC-MS.
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| Animal Protocol |
A typical in vivo animal experiment for Amlodipine-1,1,2,2-d4 maleate is a pharmacokinetic study in beagle dogs. Use male beagle dogs (10-12 kg, n=4-6). Administer a single oral dose of unlabeled Amlodipine maleate (5 mg/animal) via a gavage tube. Collect blood samples (approx. 2 mL) from the jugular vein at various time points (0, 0.5, 1, 1.5, 2, 3, 4, 6, 8, 12, 24, 36, 48, 72 hours). Collect the blood into heparinized tubes and centrifuge to obtain plasma. Store plasma samples at -80degC until analysis. For bioanalysis, spike an aliquot (50 uL) of each plasma sample with a fixed amount of the internal standard (Amlodipine-1,1,2,2-d4 maleate). Extract the samples using protein precipitation or liquid-liquid extraction. Analyze the extracted samples by LC-MS/MS. Calculate the pharmacokinetic parameters including peak plasma concentration (Cmax), time to reach Cmax (Tmax), area under the curve (AUC), elimination half-life (t½), and oral bioavailability (F) using non-compartmental analysis software (e.g., Phoenix WinNonlin).
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| ADME/Pharmacokinetics |
The pharmacokinetics of Amlodipine-1,1,2,2-d4 maleate are identical to its non-labeled parent. Amlodipine is characterized by its slow absorption, high oral bioavailability (60-80%), and very long elimination half-life (approximately 30-50 hours in humans). This long half-life is attributed to its high lipophilicity, which allows for extensive tissue distribution, and its low hepatic clearance. The volume of distribution is very large (approx. 20 L/kg), indicating extensive binding to tissues. It is extensively metabolized in the liver by CYP3A4 to inactive pyridine metabolites, but with a low extraction ratio. Approximately 10% of the parent drug is excreted unchanged in the urine. The long half-life allows for once-daily dosing in the clinic. The deuterated version (Amlodipine-d4) is used as an internal standard in mass spectrometry to provide highly accurate measurements of these PK parameters, accounting for variability in sample extraction and instrument response.
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| Toxicity/Toxicokinetics |
Amlodipine-1,1,2,2-d4 maleate is not intended for human use and serves only as an analytical research standard. The toxicity profile of its unlabeled parent, Amlodipine maleate, is well-established. Amlodipine is generally well-tolerated, with the most common adverse effects being peripheral edema (fluid accumulation in the lower legs), dizziness, flushing, and palpitations. These effects are dose-dependent and result from the drug‘s vasodilatory action. The oral LD₅0 for Amlodipine is >1000 mg/kg in rats and mice, indicating low acute toxicity. In chronic toxicology studies, high doses in dogs have been associated with gingival hyperplasia. No genotoxicity or carcinogenicity has been observed. For laboratory handling, standard safety precautions (gloves, goggles) should be used, and the compound should be stored at 4degC in a sealed container, protected from moisture.
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| References |
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| Additional Infomation |
Amlodipine-1,1,2,2-d4 maleate is the stable isotope-labeled (deuterated) version of the widely prescribed antihypertensive drug Amlodipine. It is intended for research use only and serves as a critical internal standard for the accurate quantification of Amlodipine in biological samples by LC-MS/MS. Amlodipine is a long-acting dihydropyridine calcium channel blocker that inhibits L-type calcium channels, leading to peripheral vasodilation and reduced blood pressure. It is used clinically to treat hypertension and chronic stable angina. Its slow onset and long duration of action make it a first-line agent for these conditions. Due to its long half-life, it is a common candidate for bioequivalence studies required for generic drug approval. The deuterated standard is essential for precise pharmacokinetic profiling in these studies.
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| Molecular Formula |
C24H25D4CLN2O9
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|---|---|
| Molecular Weight |
528.97
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| Exact Mass |
528.181
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| CAS # |
1185246-15-4
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| Related CAS # |
Amlodipine;88150-42-9
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| PubChem CID |
45358916
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| Appearance |
Light brown to brown solid powder
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| Melting Point |
163-174°C
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| LogP |
3.007
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
11
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| Rotatable Bond Count |
12
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| Heavy Atom Count |
36
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| Complexity |
766
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| Defined Atom Stereocenter Count |
0
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| SMILES |
[2H]C([2H])(C([2H])([2H])OCC1=C(C(C(=C(N1)C)C(=O)OC)C2=CC=CC=C2Cl)C(=O)OCC)N.C(=C\C(=O)O)\C(=O)O
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| InChi Key |
TZNOWAJJWCGILX-IBTKSTAFSA-N
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| InChi Code |
InChI=1S/C20H25ClN2O5.C4H4O4/c1-4-28-20(25)18-15(11-27-10-9-22)23-12(2)16(19(24)26-3)17(18)13-7-5-6-8-14(13)21;5-3(6)1-2-4(7)8/h5-8,17,23H,4,9-11,22H2,1-3H3;1-2H,(H,5,6)(H,7,8)/b;2-1-/i9D2,10D2;
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| Chemical Name |
(Z)-but-2-enedioic acid;3-O-ethyl 5-O-methyl 2-[(2-amino-1,1,2,2-tetradeuterioethoxy)methyl]-4-(2-chlorophenyl)-6-methyl-1,4-dihydropyridine-3,5-dicarboxylate
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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 | 1.8905 mL | 9.4523 mL | 18.9047 mL | |
| 5 mM | 0.3781 mL | 1.8905 mL | 3.7809 mL | |
| 10 mM | 0.1890 mL | 0.9452 mL | 1.8905 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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