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| 1mg |
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| Other Sizes |
| Targets |
Ivacaftor-d4 targets the CFTR protein, specifically the G551D-CFTR and other gating mutants. It binds to the CFTR channel at a site that enhances the open probability (potentiator activity), increasing chloride ion transport across the epithelial cell membrane. Ivacaftor-d4 retains the same target specificity as the unlabeled drug, but is used primarily as an analytical standard.
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| ln Vitro |
Ivacaftor (10 μM) boosted ABCB4-G535D's PC secretory activity by three times, ABCB4-G536R's by 13.7 times, ABCB4-S1076C's by 6.7 times, ABCB4-S1176L's by 9.4 times, and ABCB4-G1178S's by 5.7 times. The functional deficiencies of ABCB4 mutants are addressed by ivacaftor [1]. When comparing W1282X-expressing cells to R1162X CFTR cells, ivacaftor (10 μM) dramatically boosted CFTR activity [2]. Against 160 investigated targets, including the GABAA benzodiazepine receptor, ivacaftor had no discernible action. Ivacaftor is ten times more powerful than F508del HBEs and enhances chloride secretion with an EC50 value of 0.236 ± 0.200 μM [3]. Ivacaftor raises CFTR channel open probability (Po) in F508del processing mutation and G551D gating mutation in recombinant cells. In temperature-corrected F508del-FRT cells with an EC50 of 25 nM, ivacaftor enhances forskolin-stimulated IT by about six times [4].
Ivacaftor-d4 is not used in activity assays; however, unlabeled Ivacaftor potentiates CFTR activity in cell-free patch-clamp recordings using purified CFTR protein reconstituted into lipid bilayers. The compound increases channel open probability without affecting conductance. The d4 label does not alter the pharmacological activity, but the labeled version is used for internal standardization, not for efficacy measurements. |
| ln Vivo |
In rats, ivacaftor (1–200 mg/kg) has a good oral bioavailability [3].
In cell-based assays using Fischer rat thyroid (FRT) cells expressing G551D-CFTR, unlabeled Ivacaftor (0.1-10 uM) increases CFTR-mediated chloride transport measured by Ussing chamber or fluorescence-based membrane potential assays. Ivacaftor-d4 is not added to these functional assays; instead, it is used as an internal standard to quantify drug concentrations in cell lysates or media by LC-MS/MS. |
| Enzyme Assay |
For analytical method development, Ivacaftor-d4 is used as an internal standard in cell-free matrices. A calibration curve is prepared by spiking known concentrations of unlabeled Ivacaftor into blank plasma or buffer, along with a fixed concentration of Ivacaftor-d4. Samples are extracted by protein precipitation or solid-phase extraction and analyzed by LC-MS/MS. The peak area ratio (unlabeled/labeled) is used to quantify Ivacaftor concentration. No enzyme/receptor binding is involved.
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| Cell Assay |
Cells expressing CFTR (e.g., primary human bronchial epithelial cells from CF patients) are treated with unlabeled Ivacaftor (1-10 uM) for 1-24 hours. At the end of the experiment, cells are lysed, and Ivacaftor concentrations are quantified by LC-MS/MS using Ivacaftor-d4 as the internal standard. This allows measurement of intracellular drug accumulation. Separately, cell lysates are used for Western blot analysis of CFTR expression, but the d4 standard is not used in those assays.
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| Animal Protocol |
In animal studies, Ivacaftor-d4 is co-administered with unlabeled Ivacaftor as a tracer, or used as an internal standard in bioanalysis. Mice or rats receive Ivacaftor (10-50 mg/kg) orally. Blood samples are collected at multiple time points. Plasma is processed and analyzed by LC-MS/MS with Ivacaftor-d4 as the internal standard to quantify parent drug levels. Pharmacokinetic parameters (Cmax, Tmax, AUC, half-life) are calculated. The d4-labeled compound itself is not dosed alone for efficacy.
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| ADME/Pharmacokinetics |
Ivacaftor-d4 has identical physicochemical properties to unlabeled Ivacaftor. Ivacaftor is highly protein-bound (>99%) and is metabolized primarily by CYP3A4. The half-life in humans is approximately 12 hours. In rodents, the half-life is shorter (2-4 hours). The deuterium label does not alter the PK profile, making Ivacaftor-d4 an ideal internal standard. It is stable in plasma and during sample processing. The compound is stored at -20degC.
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| Toxicity/Toxicokinetics |
Ivacaftor-d4 is used in minute quantities as an analytical standard and does not contribute to toxicity. The unlabeled Ivacaftor is well-tolerated in patients with cystic fibrosis. Common adverse effects include headache, upper respiratory tract infection, abdominal pain, and diarrhea. More serious but rare events include elevated liver enzymes and cataracts. The deuterium label does not change the safety profile. No additional toxicity is expected.
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| References |
[1]. Delaunay JL, et al. Functional defect of variants in the adenosine triphosphate-binding sites of ABCB4 and their rescue by the cystic fibrosis transmembrane conductance regulator potentiator, ivacaftor (VX-770). Hepatology. 2017 Feb;65(2):560-570
[2]. Mutyam V, et al. Therapeutic benefit observed with the CFTR potentiator, ivacaftor, in a CF patient homozygous for the W1282X CFTR nonsense mutation. J Cyst Fibros. 2017 Jan;16(1):24-29 [3]. Hadida S, et al. Discovery of N-(2,4-di-tert-butyl-5-hydroxyphenyl)-4-oxo-1,4-dihydroquinoline-3-carboxamide (VX-770, ivacaftor), a potent and orally bioavailable CFTR potentiator. J Med Chem. 2014 Dec 11;57(23):9776-9 [4]. Van Goor F, et al. Rescue of CF airway epithelial cell function in vitro by a CFTR potentiator, VX-770. Proc Natl Acad Sci U S A. 2009 Nov 3;106(44):18825-30. |
| Additional Infomation |
Ivacaftor (VX-770) is an FDA-approved drug (Kalydeco®) for the treatment of cystic fibrosis in patients with specific CFTR gating mutations (e.g., G551D, G1244E, G1349D). It is also used in combination with lumacaftor, tezacaftor, or elexacaftor. Ivacaftor-d4 is a research-grade stable isotope-labeled internal standard intended for non-clinical analytical use only. It is not for human therapeutic use. The product is supplied as a powder with high isotopic purity.
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| Molecular Formula |
C24H24D4N2O3
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|---|---|
| Molecular Weight |
396.52
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| Related CAS # |
Ivacaftor;873054-44-5;Ivacaftor-d9;1413431-07-8;Ivacaftor benzenesulfonate;1134822-09-5;Ivacaftor hydrate;1134822-07-3;Ivacaftor-d19;1413431-22-7;Ivacaftor-d18;1413431-05-6
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| Appearance |
Typically exists as solid at room temperature
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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 | 2.5219 mL | 12.6097 mL | 25.2194 mL | |
| 5 mM | 0.5044 mL | 2.5219 mL | 5.0439 mL | |
| 10 mM | 0.2522 mL | 1.2610 mL | 2.5219 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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