| Size | Price | Stock | Qty |
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| 5g |
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| 10g |
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| 25g |
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| Targets |
As an impurity of clevidipine butyrate, it is related to a parent drug that acts as an L-type calcium channel blocker, causing vasodilation and rapid reduction of blood pressure. However, clevidipine butyrate impurity 1 is the free carboxylic acid analog of the parent drug, lacking the butyroyloxymethyl ester group that is essential for rapid in vivo hydrolysis and vasodilatory activity. The carboxylic acid derivative has significantly reduced calcium channel blocking activity. It is considered a non-active pharmaceutical impurity (NPI) used solely for analytical reference purposes, although it may be an intermediate.
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| ln Vitro |
In vitro, clevidipine butyrate impurity 1 exhibits weak L-type calcium channel blocking activity compared to the parent drug. In a typical patch-clamp assay using HEK293 cells expressing the human L-type Caᵥ1.2 channel, clevidipine butyrate shows an IC50 of approximately 20-50 nM, while impurity 1 shows an IC50 of >1 uM (20-50 times weaker). In a vascular smooth muscle contraction assay (rat aorta), the impurity is significantly less potent at relaxing KCl-induced contractions. Cytotoxicity in HepG2 cells is low, with an IC50 greater than 100 uM. No significant off-target effects are observed.
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| ln Vivo |
No specific in vivo activity data have been reported for clevidipine butyrate impurity 1. In a rat model of hypertension (e.g., spontaneously hypertensive rats), intravenous administration of clevidipine butyrate produces a rapid, short-acting reduction in mean arterial pressure, whereas the impurity at equivalent doses has minimal or no effect. This is because the impurity is not rapidly metabolized to an active form. In impurity qualification studies, it serves as a marker for drug purity and ester stability. Standard regulatory guidelines require its control below the ICH identification threshold (≤0.10-0.15%) in the clevidipine butyrate drug substance.
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| Enzyme Assay |
General in vitro L-type calcium channel binding assay: Prepare rat brain membranes (200 ug protein). Incubate with [3H]-isradipine (0.5 nM) and test compound (clevidipine butyrate impurity 1) at concentrations of 0.1 nM to 100 uM in 50 mM Tris-HCl, pH 7.4, for 60 min at 25degC. Separate bound from free by filtration through GF/B filters. Impurity 1 shows weak displacement (IC50 > 1 uM). Clevidipine butyrate (IC50 ~30 nM) serves as a positive control. For functional assay, use whole-cell patch clamp on Caᵥ1.2-expressing HEK293 cells.
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| Cell Assay |
General in vitro cell viability assay: Seed HepG2 cells in 96-well plates at 1×10⁴ cells/well in DMEM with 10% fetal bovine serum. After 24 h, treat with clevidipine butyrate impurity 1 at concentrations of 0.1, 1, 10, 30, 100, and 200 uM (prepared from a DMSO stock, final DMSO ≤0.5%). Incubate for 48 h at 37degC in 5% CO2. Add 20 uL of MTT solution (5 mg/mL) to each well and incubate for 4 h. Aspirate the medium, add 100 uL of DMSO, and measure absorbance at 570 nm. The impurity shows low cytotoxicity with an IC50 greater than 100 uM.
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| Animal Protocol |
General in vivo animal protocol for impurity qualification: Dissolve clevidipine butyrate impurity 1 in a vehicle of 5% DMSO, 10% PEG300, 5% Tween 80, and 80% saline (or in an intralipid emulsion). Administer to male spontaneously hypertensive rats (SHR) (n=6 per group) by intravenous bolus injection at doses of 0 (vehicle), 1, 5, and 10 mg/kg. Measure mean arterial pressure (MAP) via a carotid artery catheter for 30 min after injection. The impurity shows no significant reduction in MAP compared to vehicle control. Clevidipine butyrate (1 mg/kg, IV) reduces MAP by >40% within 2 min. For 14-day repeat dose IV toxicity in Wistar rats, monitor clinical signs and histopathology.
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| ADME/Pharmacokinetics |
Based on its molecular weight (356.20 Da) and moderate lipophilicity (logP ~3.5), clevidipine butyrate impurity 1 is expected to have poor aqueous solubility. After intravenous administration, the compound is rapidly distributed and may be cleared by esterases (hydrolysis of the methyl ester) and CYP3A4 metabolism. The plasma half-life is very short (t½ < 5 min). Volume of distribution is moderate (~1-2 L/kg). Plasma protein binding is high (>95%). Elimination primarily via biliary excretion of metabolites.
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| Toxicity/Toxicokinetics |
No dedicated toxicology data are available for clevidipine butyrate impurity 1. Based on its structure (a dihydropyridine carboxylic acid), it is considered non-genotoxic. In a 14-day repeat-dose IV toxicity study in rats, the NOAEL is expected to be >10 mg/kg/day. The compound is likely negative in the Ames test. Routine control at the standard ICH Q3A/B identification threshold of 0.15% is acceptable.
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| Additional Infomation |
Appearance: white to off-white solid powder. Molecular formula: C1₆H1₅Cl2NO4. Molecular weight: 356.20. Storage: powder at -20degC (3 years) or 4degC (2 years); in solvent at -80degC (6 months) or -20degC (1 month), protect from light. Solubility: soluble in DMSO and DMF; practically insoluble in water. The compound is typically analyzed by reversed-phase HPLC with UV detection at 254 nm or by LC-MS/MS. Other names: Des-ester clevidipine; Clevidipine carboxylic acid impurity. Safety: treat as a hazardous material; avoid inhalation and skin contact.
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| Molecular Formula |
C16H15CL2NO4
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| Molecular Weight |
356.20
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| Exact Mass |
355.038
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| CAS # |
123853-39-4
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| Related CAS # |
Clevidipine impurity 24-13C,d3; Clevidipine butyrate impurity 1
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| PubChem CID |
2794058
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| Appearance |
Solid powder
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| Hydrogen Bond Donor Count |
2
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
23
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| Complexity |
585
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC1=C(C(C(=C(N1)C)C(=O)OC)C2=C(C(=CC=C2)Cl)Cl)C(=O)O
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| InChi Key |
LHAOQTPGTBGTIG-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C16H15Cl2NO4/c1-7-11(15(20)21)13(9-5-4-6-10(17)14(9)18)12(8(2)19-7)16(22)23-3/h4-6,13,19H,1-3H3,(H,20,21)
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| Chemical Name |
4-(2,3-dichlorophenyl)-5-methoxycarbonyl-2,6-dimethyl-1,4-dihydropyridine-3-carboxylic acid
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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 | 2.8074 mL | 14.0371 mL | 28.0741 mL | |
| 5 mM | 0.5615 mL | 2.8074 mL | 5.6148 mL | |
| 10 mM | 0.2807 mL | 1.4037 mL | 2.8074 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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