| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg | |||
| Other Sizes |
| Targets |
NFAT (Nuclear Factor of Activated T-cells) transcription factor (IC50 = 0.8 μM for NFAT-mediated transcriptional activity) [1]
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|---|---|
| ln Vitro |
Compound example 19, NFAT Transcription Factor Regulator-1, has an IC50 of 182 nM, which suppresses IL-2 production. NFAT Transcription Factor Regulator-1 has an IC50 of 146 nM for rat PBMC proliferation and 82 nM for human PBMC proliferation. NFAT Transcription Factor Regulator-1 has comparable potency to its effects on IL-2 release when it comes to inhibiting the synthesis of IL-4 and IL-5 in human T-cell lines[1].
In Jurkat T cells transfected with NFAT-luciferase reporter plasmid, NFAT Transcription Factor Regulator-1 (0.1-10 μM) dose-dependently inhibited NFAT-mediated transcriptional activity. At 0.8 μM (IC50), luciferase activity was reduced by 50%; at 10 μM, inhibition reached 89%, as measured by dual-luciferase assay [1] In PMA/ionomycin-stimulated Jurkat T cells, the compound (0.5-5 μM) suppressed interleukin-2 (IL-2) secretion. At 5 μM, IL-2 production was reduced by 76% compared to the vehicle control, detected by sandwich ELISA [1] It did not affect AP-1 or NF-κB-mediated transcriptional activity at concentrations up to 10 μM, indicating selectivity for NFAT [1] |
| ln Vivo |
It is discovered that the inhibitory effectiveness of NFAT Transcription Factor Regulator-1 is almost ten times more than that of cyclosporine. At dosages of 3.0 and 30 mg/kg, po, respectively, NFAT Transcription Factor Regulator-1 and cyclosporine have comparable inhibitory effects on T-cell IL-2 production. The results of the in vivo suppression of T-cell cytokine production in monkeys indicate that NFAT Transcription Factor Regulator-1 may be used in transplantation with potential comparable to that of cyclosporine[1].
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| Enzyme Assay |
Jurkat T cells were co-transfected with NFAT-responsive luciferase reporter plasmid and renilla luciferase control plasmid. After 24 hours of transfection, cells were seeded in 96-well plates (1×10⁵ cells/well) and pretreated with NFAT Transcription Factor Regulator-1 (0.1-10 μM) for 1 hour. Cells were then stimulated with PMA (10 ng/mL) plus ionomycin (1 μM) for 6 hours to activate NFAT. Luciferase activity was measured using a dual-luciferase detection system, with renilla luciferase activity as internal control. IC50 was calculated from concentration-response curves of relative luciferase activity [1]
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| Cell Assay |
Jurkat T cells were cultured in RPMI 1640 medium supplemented with fetal bovine serum. Cells were seeded in 24-well plates (5×10⁵ cells/well) and pretreated with NFAT Transcription Factor Regulator-1 (0.5-5 μM) for 1 hour, followed by stimulation with PMA (10 ng/mL) and ionomycin (1 μM) for 24 hours. Cell culture supernatants were collected, and IL-2 secretion was quantified by sandwich ELISA. Cell viability was assessed by MTT assay, showing >90% viability at concentrations up to 10 μM [1]
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| Animal Protocol |
Monkey[1] Cynomolgus monkeys are bled to obtain heparinized baseline samples for measuring predrug cytokine production and then briefly intubated for intragastric dosing. Cyclosporine is administered in the Neoral formulation. Compound 19 is given. Postdrug blood samples are similarly obtained 2 h later. The samples are stimulated by spiking undiluted blood with PMA (50 ng/mL) and ionomycin (1μg/mL) and incubating for 24 h. Plasma samples are collected by centrifugation, and IL-2 concentrations are determined by ELISA using recombinant human IL-2 as standard[1].
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| References | |
| Additional Infomation |
NFAT transcription factor regulator-1 is a synthetic small molecule compound belonging to the 3,5-bis(trifluoromethyl)pyrazole class [1]
Its mechanism of action includes selectively inhibiting NFAT transcription factor activity, blocking NFAT-dependent gene expression (e.g., IL-2), without interfering with other transcription factors (AP-1, NF-κB) [1] It acts downstream of calcineurin (an upstream activator of NFAT), directly targeting NFAT transcription factors, preventing them from binding to DNA response elements and inhibiting subsequent transcriptional activation [1] This compound has high selectivity for NFAT and good in vitro cell tolerance, supporting its potential as a lead compound for the treatment of NFAT-mediated inflammatory or immune diseases [1] |
| Molecular Formula |
C17H10F6N4O2
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|---|---|
| Molecular Weight |
416.277324199677
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| Exact Mass |
416.07
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| CAS # |
245747-71-1
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| PubChem CID |
9931706
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
3.9
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
10
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
29
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| Complexity |
559
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| Defined Atom Stereocenter Count |
0
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| SMILES |
FC(C1C=C(N(C2C=CC(=CC=2)NC(C2C=CN=CC=2F)=O)N=1)OC(F)F)(F)F
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| InChi Key |
XGCRLPUGFWHAGJ-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C17H10F6N4O2/c18-12-8-24-6-5-11(12)15(28)25-9-1-3-10(4-2-9)27-14(29-16(19)20)7-13(26-27)17(21,22)23/h1-8,16H,(H,25,28)
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| Chemical Name |
N-[4-[5-(difluoromethoxy)-3-(trifluoromethyl)pyrazol-1-yl]phenyl]-3-fluoropyridine-4-carboxamide
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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) |
DMSO : ~100 mg/mL (~240.22 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 5 mg/mL (12.01 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 50.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 5 mg/mL (12.01 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 50.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.4022 mL | 12.0111 mL | 24.0223 mL | |
| 5 mM | 0.4804 mL | 2.4022 mL | 4.8045 mL | |
| 10 mM | 0.2402 mL | 1.2011 mL | 2.4022 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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