PRN694
Based on 3 publication(s) in Google Scholar
PRN694 is an irreversible, highly selective and potent covalent interleukin-2-inducible T-cell kinase (ITK) and resting lymphocyte kinase (RLK) dual inhibitor with IC50s of 0.3 nM and 1.4 nM, respectively. PRN694 exhibits extended target residence time on ITK and RLK, enabling durable attenuation of effector cells in vitro and in vivo.
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- Pureza: ≥99.0%
- No. CAS: 1575818-46-0
- Fòrmula: C28H35F2N5O2S
- Peso molecular:543.67
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Almacenamiento:Powder -20°C, 3 years , 4°C, 2 years ; In solvent -80°C, 2 years , -20°C, 1 year
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Actividad biológica
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Chemical Information
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Solvente y solubilidad
- Protocolo
- Pureza y Documentación
- Referencias
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Help & FAQs
Publications Citing Use of MedChemExpress (MCE) PRN694
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Actividad biológica
IC50: 0.3 nM (ITK), 1.4 nM (RLK), 3.3 nM (TEC), 17 nM (BTK), 17 nM (BMX), 30 nM (JAK3), 125 nM (BLK)[1]
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Cell Line
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Type | Value | Description | References |
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| Jurkat | EC50 |
157 nM
Compound: 9; PRN694
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Inhibition of ITK in anti-CD3/CD28 beads-stimulated human Jurkat cells assessed as reduction in Plcgamma1 phosphorylation incubated for 2 hrs followed by stimulation with anti-CD3/CD28 beads by Western blot analysis
Inhibition of ITK in anti-CD3/CD28 beads-stimulated human Jurkat cells assessed as reduction in Plcgamma1 phosphorylation incubated for 2 hrs followed by stimulation with anti-CD3/CD28 beads by Western blot analysis
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[PMID: 31830635] |
PRN694 inhibits TEC, BTK, BMX, BLK, JAK3 with IC50s of 3.3, 17, 17, 125, 30 nM, respectively[1].
Immunoblot analysis of TCR activation pathways reveales that PRN694 blocks activation or nuclear translocation of NFAT1, JunB, pIκBα, and pERK. Results reveal inhibition of Ca2+ signaling with PRN694 at all concentrations above 1 nM. PRN694 significantly attenuates NK cell FcR-induced killing at concentrations exceeding 0.37 μM[1].
Day 6 flow cytometry analysis reveals that PRN694 significantly inhibits the anti-CD3/CD28-induced proliferation of both CD4 and CD8 T-cells (p<0.01)[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Colitis studies show reduced numbers of CD4+ T cells present in the colonic epithelium of PRN694-treated mice compare with controls[2].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Chemical Information
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No. CAS 1575818-46-0
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Appearance Solid
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Peso molecular 543.67
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Fòrmula C28H35F2N5O2S
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Color Off-white to light brown
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SMILES
O=C(C1=CC=C(C(F)F)S1)NC2=NC3=CC(CN[C@@H](C)C(C)(C)C)=CC=C3N2C[C@@H]4N(C(C=C)=O)CCC4
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Envío
Room temperature in continental US; may vary elsewhere.
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Almacenamiento
Powder -20°C 3 years 4°C 2 years In solvent -80°C 2 years -20°C 1 year
Publications (3)
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Journal Impact Factor
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Most Recent
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Sci Signal
TNF-α signals through ITK-Akt-mTOR to drive CD4+ T cell metabolic reprogramming, which is dysregulated in rheumatoid arthritis. [Abstract]2024 Apr 23;17(833):eadg5678. PMID: 38652761 -
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Solvente y solubilidad
DMSO : 125 mg/mL (229.92 mM; Need ultrasonic and warming; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
Select the appropriate dissolution method based on your experimental animal and administration route.
- For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
- To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for In Vivo experiments, it is recommended to prepare freshly and use it on the same day.
- The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.
Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
Solubility: ≥ 2.08 mg/mL (3.83 mM); Clear solution
This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)
Solubility: ≥ 2.08 mg/mL (3.83 mM); Clear solution
This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
Please enter the basic information of animal experiments:
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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
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%DMSO +
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
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%+
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+%Tween-80 + +
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%Saline +
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Working solution concentration: 0.22 mg/mL
Method for preparing stock solution: mg drug dissolved in μL DMSO. Stock solution concentration: mg/mL.
1. Take μL DMSO stock solution;
2. Add μL .
μL , mix evenly;
3. Then add μL Tween 80, mix evenly;
4. Then add μL
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Protocolo
Recombinant ITK at a final concentration of 0.5 μM in 50 mM Hepes, pH 7.5, 10 mM MgCl2, 0.01% Triton X-100, and 1 mM EGTA are combined with 1.5 μM PRN694 for 90 min to facilitate binding. The mixture is then diluted 50 fold to initiate dissociating of the ligand from the enzyme, and 10 μL is transferred to a Greiner 384-well black plate. Europium-coupled anti-His6 antibody is added to each well and incubated for 5 min, followed by the addition of an ITK binding fluorescent tracer. The tracer binds to ITK as a function of ligand dissociation, and binding is detected by time-resolved FRET between the europium-coupled antibody and the tracer on a plate reader. Time points acquired are 0.25, 1, 3, 6, and 24 h[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Cells are cultured in vitro at 37°C and 5% CO2 using RPMI 1640 with 10% fetal calf serum. Cells are pretreated for 30 min with PRN694 or other inhibitors and then washed two times. T-cells are then stimulated for 6 h with 1 μg/mL soluble anti-CD3 for CD69 activation, which is detected by flow cytometry, or 45 min with plate-bound anti-CD3 (10 μg/mL plating concentration) and soluble anti-CD28 (1 μg/mL) for downstream signal analysis by immunoblotting. NK cells are stimulated for 6 h with plate-bound anti-CD52 for CD107a/b activation, detected by flow cytometry, or for 45 min for downstream signal analysis by immunoblotting[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Mice are randomized by weight and sensitized with aliquots of 150 μL of 5% oxazolone in 3 parts ethanol and 1 part acetone on their shaved abdomens. Seven days after the sensitization, the mice are challenged with 10 μL of 3% oxazolone on the front and back of the right ears. The left ears are treated with the ethanol/acetone mixture. One hour prior to the challenge, the animals received either vehicle control (5% ethanol, 95% Captex 355 NP/EF, intraperitoneal injection at 5 mL/kg), 20 mg/kg PRN694 in 5% ethanol, 95% Captex (intraperitoneal injection at 5 mL/kg), or 0.5 mg/kg dexamethasone (intraperitoneal injection at 5 mL/kg). A control group of animals receive no oxazolone or drug treatment. Twenty-four hours after the oxazolone challenge, the mice are sacrificed, and a 7 mm disc is punched out of each ear and weighed to measure edema[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Pureza y Documentación
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Ficha de datos (280 KB)
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SDS (393 KB)
- English - EN (393 KB)
- Français - FR (393 KB)
- Deutsch - DE (393 KB)
- Norwegian - NO (393 KB)
- Español - ES (393 KB)
- Swedish - SV (393 KB)
- Italian - IT (393 KB)
- Korean - KR (393 KB)
- Portuguese - PT (393 KB)
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Instrucciones de manejo (2659 KB)
Referencias
[1]. Zhong Y, et al. Targeting interleukin-2-inducible T-cell kinase (ITK) and resting lymphocyte kinase (RLK) using a novel covalent inhibitor PRN694. J Biol Chem. 2015 Mar 6;290(10):5960-78. [Content Brief]
[2]. Cho HS, et al. A Small Molecule Inhibitor of ITK and RLK Impairs Th1 Differentiation and Prevents Colitis Disease Progression. J Immunol. 2015 Nov 15;195(10):4822-31. [Content Brief]
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
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| DMSO | 1 mM | 1.8394 mL | 9.1968 mL | 18.3935 mL | 45.9838 mL |
| 5 mM | 0.3679 mL | 1.8394 mL | 3.6787 mL | 9.1968 mL | |
| 10 mM | 0.1839 mL | 0.9197 mL | 1.8394 mL | 4.5984 mL | |
| 15 mM | 0.1226 mL | 0.6131 mL | 1.2262 mL | 3.0656 mL | |
| 20 mM | 0.0920 mL | 0.4598 mL | 0.9197 mL | 2.2992 mL | |
| 25 mM | 0.0736 mL | 0.3679 mL | 0.7357 mL | 1.8394 mL | |
| 30 mM | 0.0613 mL | 0.3066 mL | 0.6131 mL | 1.5328 mL | |
| 40 mM | 0.0460 mL | 0.2299 mL | 0.4598 mL | 1.1496 mL | |
| 50 mM | 0.0368 mL | 0.1839 mL | 0.3679 mL | 0.9197 mL | |
| 60 mM | 0.0307 mL | 0.1533 mL | 0.3066 mL | 0.7664 mL | |
| 80 mM | 0.0230 mL | 0.1150 mL | 0.2299 mL | 0.5748 mL | |
| 100 mM | 0.0184 mL | 0.0920 mL | 0.1839 mL | 0.4598 mL |
PRN694 Related Classifications
HY-12680 Related Classifications
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