| Size | Price | Stock | Qty |
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| 10mg |
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| Targets |
EC50: 64 nM (WT CFTR)[1]
I1421 targets the cystic fibrosis transmembrane conductance regulator (CFTR), an ABC transporter that functions as a chloride channel on the apical surface of epithelial cells. I1421 is a potentiator, meaning it enhances the channel open probability (gating) of CFTR at the cell surface. It binds allosterically (at a site distinct from the ATP-binding pocket) to both wild-type and mutant CFTR, increasing the flow of chloride ions. This corrects the defective ion transport that is the underlying cause of cystic fibrosis. I1421 targets multiple CF-causing mutants, including deltaF508 and G551D CFTR. |
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| ln Vitro |
In vitro, I1421 is a potent CFTR potentiator with an EC50 of 64 nM for wild-type CFTR currents. It allosterically activates a wide range of CF-causing mutants, such as deltaF508 and G551D CFTR. In electrophysiological assays using CFTR-expressing cells, I1421 increases CFTR-mediated chloride currents in a concentration-dependent manner. The compound activates various CF-causing mutants, demonstrating high efficacy across multiple genotypes. I1421 also shows excellent in vitro potency with minimal cytotoxicity. No specific IC50 values are reported.
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| ln Vivo |
I1421 (10 mg/kg; ip, po, sc; single dose) is rapidly absorbed in C57BL/6 mice. After three different administration routes, the maximum plasma concentrations (Cmax) observed 15 min after intraperitoneal and oral administration were 7.4 μM and 3.4 μM, respectively. The plasma level after intraperitoneal administration was higher than that after the other two administration routes [1]. The bioavailability of I1421 (3 mg/kg; iv, po; single dose) was 60% in mice after oral and intravenous injection, but the oral T1/2 was short (75 min) [1].
In vivo, I1421 exhibits good in vivo efficacy, with an oral bioavailability of 60% in mice and a corresponding half-life of 75 minutes. It has been shown to allosterically activate multiple CF-causing mutants with good in vivo potency. I1421 may have a synergistic effect when combined with other CFTR modulators. The compound‘s physical chemistry and pharmacokinetic properties are favorable. |
| Enzyme Assay |
The binding of I1421 to CFTR is measured by a combination of electrophysiological and biophysical methods. Patch-clamp electrophysiology is the gold standard for measuring CFTR channel activity. CFTR-expressing cells (e.g., CHO or BHK cells) are used in the whole-cell or excised patch configuration. I1421 is applied at graded concentrations (0.1-10,000 nM) to the bath solution, and the increase in chloride current is measured. The EC50 of 64 nM is calculated. For binding studies, a fluorescently labeled CFTR potentiator (e.g., a fluorescent analog of VX-770) can be used to measure competitive binding to CFTR by fluorescence polarization (FP) or by using a TIRF microscope. No specific KD value is reported.
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| Cell Assay |
For cellular assays, CFTR-expressing cells (e.g., CHO cells stably expressing wild-type or mutant CFTR, or patient-derived human bronchial epithelial cells) are seeded in 96-well plates. A fluorescence-based membrane potential assay (e.g., FLIPR assay) is used. Cells are loaded with a voltage-sensitive dye, and the change in fluorescence is measured after adding I1421 at graded concentrations (0.1-10,000 nM) in the presence of a chloride gradient. The EC50 is calculated from the fluorescence increase. For chloride efflux assays, cells are loaded with a chloride-sensitive dye (e.g., MQAE), and the rate of dequenching is measured. For validation, forskolin (10 uM) is used to activate CFTR.
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| Animal Protocol |
For in vivo evaluation of CFTR potentiation, a CFTR-mediated intestinal current measurement (ICM) assay is used. mice (wild-type or CFTR mutant mice, e.g., CFTR-deltaF508 mice) are used. I1421 is administered orally by gavage at doses of 10-100 mg/kg. After 1-4 hours, the mice are euthanized, and segments of the intestine (e.g., jejunum, ileum) are harvested. The tissue is mounted in Ussing chambers, and the short-circuit current (Isc) is measured in response to stimulation with a cAMP activator (e.g., forskolin). The increase in Isc is measured. The sweat chloride test is another in vivo assay: the compound is administered, and sweat is collected; chloride concentration is measured by coulometric titration. For PK studies, blood is collected at various time points, and plasma I1421 concentrations are measured by LC-MS/MS. In mice, oral bioavailability is 60% and half-life is 75 min.
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| ADME/Pharmacokinetics |
I1421 (C2₆H1₈F4N4O4S, MW = 558.50, purity 99.15%, CAS 1638866-84-4) is a solid powder. For storage, the powder should be kept at -20degC for up to 3 years, sealed and protected from light. For in vitro use, stock solutions in DMSO (50 mg/mL, 89.53 mM) can be stored at -80degC for up to 6 months or at -20degC for 1 month. For in vivo oral administration, it can be formulated in 0.5% methylcellulose/0.1% Tween-80 or in 10% DMSO / 40% PEG300 / 5% Tween-80 / 45% saline. In mice, oral bioavailability is 60%, and half-life is 75 minutes. No other PK parameters (Cmax, Tmax, AUC) are reported.
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| Toxicity/Toxicokinetics |
No specific toxicity data for I1421 are reported. As a research-grade CFTR potentiator, it is not intended for human or veterinary use. Standard laboratory safety precautions for handling chemicals should be followed. CFTR potentiators as a class (e.g., ivacaftor) have been approved for clinical use and have a well-established safety profile. However, no formal toxicology studies (LD50, target organ toxicity) are available for I1421.
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| References | |
| Additional Infomation |
I1421 is a research-grade, small-molecule CFTR potentiator. Cystic fibrosis (CF) is a genetic disorder caused by mutations in the CFTR gene, leading to a defective CFTR chloride channel. More than 2,000 mutations have been identified, with deltaF508 (a deletion of phenylalanine at position 508) being the most common. G551D is a gating mutation (impaired channel opening). Potentiators are drugs that increase the open probability of CFTR channels at the cell surface. I1421 allosterically activates a wide range of CF-causing mutants. I1421 was discovered by researchers at the University of California, San Francisco (UCSF) and is described in the literature (e.g., Yeh et al., Cell Chemical Biology). The compound has favorable physico-chemical properties and pharmacokinetics. I1421 is for research use only and has not received regulatory approval.
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| Molecular Formula |
C18H18FN3O3
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| Molecular Weight |
343.35
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| Appearance |
White to off-white solid powder
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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 | 2.9125 mL | 14.5624 mL | 29.1248 mL | |
| 5 mM | 0.5825 mL | 2.9125 mL | 5.8250 mL | |
| 10 mM | 0.2912 mL | 1.4562 mL | 2.9125 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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