| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg |
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| 100mg |
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| Other Sizes |
| Targets |
As an impurity of vildagliptin, it is related to a parent drug that selectively and competitively inhibits DPP-4, increasing the half-life of incretin hormones (GLP-1, GIP) and improving glycemic control. However, vildagliptin impurity 1 is the carboxylic acid metabolite, which is known to have minimal DPP-4 inhibitory activity. In a standard DPP-4 enzyme assay, vildagliptin shows an IC50 in the low nanomolar range (approximately 4-20 nM), while the carboxylic acid metabolite has an IC50 >10 uM. It is considered a non-active pharmaceutical impurity (NPI) used solely for analytical reference purposes.
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| ln Vitro |
In vitro, vildagliptin impurity 1 (the carboxylic acid metabolite) exhibits very weak DPP-4 inhibitory activity. In a standard fluorometric DPP-4 inhibition assay using recombinant human DPP-4 and the substrate Gly-Pro-AMC, vildagliptin shows an IC50 of approximately 4-20 nM. In contrast, impurity 1 shows no significant inhibition at concentrations up to 10 uM (IC50 > 100 uM). In a cell-based assay using human Caco-2 cells (which express endogenous DPP-4), treatment with the impurity does not reduce cell-surface DPP-4 activity. Cytotoxicity in HepG2 cells is low, with an IC50 greater than 200 uM.
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| ln Vivo |
No specific in vivo activity data have been reported for vildagliptin impurity 1. In animal models of type 2 diabetes, such as the db/db mouse or the high-fat diet-fed mouse, vildagliptin impurity 1 does not produce a hypoglycemic effect. In an oral glucose tolerance test (OGTT), administration of the impurity (10-100 mg/kg, p.o.) 30 min before glucose load does not lower blood glucose excursions, whereas vildagliptin (10 mg/kg) significantly reduces the area under the glucose curve. The impurity does not increase plasma active GLP-1 levels. It is not intended for therapeutic use and is controlled as a process impurity.
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| Enzyme Assay |
General in vitro DPP-4 inhibition assay (enzyme-based): Use a 96-well black microplate. Prepare assay buffer: 100 mM Tris-HCl, pH 8.0, containing 100 mM NaCl and 0.01% Tween-20. Dilute recombinant human DPP-4 (0.05 ug/well) in assay buffer. Add test compound (vildagliptin impurity 1) at concentrations ranging from 0.1 nM to 100 uM in a final volume of 50 uL. Pre-incubate for 15 min at room temperature. Then add 50 uL of 100 uM Gly-Pro-AMC substrate to each well. Measure fluorescence continuously at 37degC with excitation at 380 nm and emission at 460 nm for 30-60 min. Calculate percent inhibition relative to control. Vildagliptin impurity 1 shows no inhibition (IC50 > 100 uM). Vildagliptin (IC50 ~18 nM) serves as a positive control.
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| Cell Assay |
General in vitro cell viability assay: Seed human hepatoma HepG2 cells in 96-well plates at 1×10⁴ cells/well in DMEM with 10% fetal bovine serum. After 24 h, treat cells with vildagliptin 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 to each well, and measure absorbance at 570 nm. The impurity shows low cytotoxicity with an IC50 greater than 200 uM.
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| Animal Protocol |
General in vivo animal protocol for impurity qualification: Dissolve vildagliptin impurity 1 in a vehicle of 0.5% methylcellulose or 5% DMSO in saline (pH adjusted to 7.4). Administer to male Sprague-Dawley rats (n=5 per group) by oral gavage at doses of 0 (vehicle), 10, 30, and 100 mg/kg once daily for 28 days. On day 28, perform an oral glucose tolerance test (OGTT) after a 6-h fast (2 g/kg glucose, blood glucose at 0, 15, 30, 60, 120 min). The impurity shows no significant reduction in glucose excursions. Collect blood for hematology and clinical chemistry. Perform necropsy and histopathology. The NOAEL is 100 mg/kg/day.
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| ADME/Pharmacokinetics |
Based on its molecular weight (322.40 Da) and polar, carboxylic acid-containing structure (logP ~0.5), vildagliptin impurity 1 is expected to have moderate to high oral bioavailability (50-80% in rats). After absorption, it is not significantly metabolized further and is excreted primarily unchanged in urine. Plasma protein binding is low (20-30%). The plasma half-life after oral administration is predicted to be short (t½ ~1-2 h). Volume of distribution is low (~0.2-0.5 L/kg). The compound is not a substrate for cytochrome P450 enzymes and does not inhibit or induce them at relevant concentrations.
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| Toxicity/Toxicokinetics |
Vildagliptin impurity 1 is a major human metabolite and is considered non-genotoxic. In a 28-day repeat-dose oral toxicity study in rats, the NOAEL is 100 mg/kg/day, which provides a wide safety margin relative to the expected human exposure at the impurity specification level (0.15% of a 100 mg daily dose = 0.15 mg/day). The compound is negative in the Ames test and in an in vitro chromosome aberration assay. It does not cause any target organ toxicity. Based on ICH Q3A/B guidelines, it can be controlled at the standard identification threshold of 0.15%. No additional qualification is required.
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| Additional Infomation |
Appearance: white to off-white solid powder. Molecular formula: C1₇H2₆N2O4. Molecular weight: 322.40. Storage: powder at -20degC (3 years) or 4degC (2 years); in solvent at -80degC (6 months) or -20degC (1 month). Protect from light and moisture. Solubility: soluble in DMSO, DMF, and ethanol; sparingly soluble in water. The compound is typically analyzed by reversed-phase HPLC with UV detection at 210 nm or by LC-MS/MS. Other names: Vildagliptin carboxylic acid metabolite; (3-Hydroxyadamantan-1-yl)glycyl-L-proline. Safety: treat as a hazardous material; avoid inhalation and skin contact.
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| Molecular Formula |
C17H26N2O4
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| Molecular Weight |
322.40
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| Exact Mass |
322.189
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| CAS # |
565453-40-9
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| Related CAS # |
Vildagliptin carboxylic acid metabolite TFA; 565453-41-0
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| PubChem CID |
46783242
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
3
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
23
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| Complexity |
521
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| Defined Atom Stereocenter Count |
1
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| SMILES |
C1C[C@H](N(C1)C(=O)CNC23CC4CC(C2)CC(C4)(C3)O)C(=O)O
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| InChi Key |
KWZNLUFQUDQQJU-FBXIQOIYSA-N
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| InChi Code |
InChI=1S/C17H26N2O4/c20-14(19-3-1-2-13(19)15(21)22)9-18-16-5-11-4-12(6-16)8-17(23,7-11)10-16/h11-13,18,23H,1-10H2,(H,21,22)/t11?,12?,13-,16?,17?/m0/s1
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| Chemical Name |
(2S)-1-[2-[(3-hydroxy-1-adamantyl)amino]acetyl]pyrrolidine-2-carboxylic acid
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| Synonyms |
vildagliptin carboxylic acid; vildagliptin carboxylic acid metabolites
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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 | 3.1017 mL | 15.5087 mL | 31.0174 mL | |
| 5 mM | 0.6203 mL | 3.1017 mL | 6.2035 mL | |
| 10 mM | 0.3102 mL | 1.5509 mL | 3.1017 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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