| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg | |||
| Other Sizes |
| Targets |
Interleukin-2-inducible T-cell kinase (ITK). GNE-9822 is a potent and highly selective ITK inhibitor that binds to the ATP-binding pocket of ITK with a Ki of 0.7 nM. The compound demonstrates exceptional selectivity over other kinases, with only six of 286 off-target kinases showing >70% inhibition when tested at 0.1 μM, which is more than 100-fold above its ITK Ki. This remarkable selectivity profile distinguishes GNE-9822 from earlier ITK inhibitors and makes it a valuable tool for dissecting ITK-specific functions in T-cell signaling without confounding off-target effects.
|
|---|---|
| ln Vitro |
GNE-9822 (compound 28) has an IC50 value of 55 nM, which inhibits PLCγ phosphorylation[1].
GNE-9822 inhibits ITK with a Ki of 0.7 nM in biochemical assays, demonstrating high affinity for the target enzyme. In cellular assays, GNE-9822 (compound 28) inhibits the phosphorylation of PLCγ, a key downstream effector of ITK-mediated TCR signaling, with an IC50 of 55 nM. The compound has an EC50 of 354.5 nM in functional cellular assays. GNE-9822 demonstrates exceptional broad kinome selectivity: when tested at 0.1 μM (>100× the ITK Ki of 0.7 nM) against a panel of 286 off-target kinases, only six kinases were inhibited >70%. This remarkable selectivity ensures that the observed cellular effects are primarily attributable to ITK inhibition rather than off-target kinase activities. |
| ln Vivo |
GNE-9822 (compound 28; po and iv; 1 and 5 mg/kg) showed good bioavailability and half-life in mice and stents [1].
In vivo, GNE-9822 (compound 28) administered via oral (po) and intravenous (iv) routes at doses of 1 and 5 mg/kg showed good bioavailability and favorable half-life in mice and rats. The compound has been evaluated in murine models of ovalbumin (OVA) and house dust mite (HDM)-induced asthma, where its oral bioavailability of 36% in mice supports twice-daily (BID) dosing regimens. In conscious guinea pigs, GNE-9822 (0.3 mg/kg; i.p.) dramatically lowers extracellular 5-HT in the frontal cortex. These in vivo studies demonstrate that GNE-9822 is effective in modulating ITK-dependent pathways in living organisms and support its utility as a tool compound for investigating the role of ITK in inflammatory disease models. |
| Enzyme Assay |
GNE-9822 ITK inhibition is assessed using in vitro kinase assays with recombinant ITK enzyme and a substrate peptide. Radiolabeled ATP or luminescent-based assays (e.g., ADP-Glo) are used to measure kinase activity, and Ki values are calculated from dose-response curves using appropriate enzyme kinetics models. Selectivity profiling is performed by testing the compound at 0.1 μM (more than 100-fold above its ITK Ki of 0.7 nM) against a panel of 286 off-target kinases in competition binding or activity assays; only kinases showing >70% inhibition are considered significant hits. Surface plasmon resonance (SPR) or isothermal titration calorimetry (ITC) may be used to directly measure binding affinity and kinetics.
|
| Cell Assay |
Cellular ITK activity is evaluated in T-cell lines or primary T cells. Cells are treated with GNE-9822 at various concentrations (typically 0.001-10 μM) for 1-4 hours prior to TCR stimulation with anti-CD3/anti-CD28 antibodies. ITK-mediated signaling is assessed by measuring PLCγ1 phosphorylation via Western blot using phospho-specific antibodies (pY783). Calcium flux is measured using fluorescent calcium indicators such as Fluo-4 AM in flow cytometry or plate-reader-based assays. Cytokine production (IL-2, IFN-γ, IL-4, IL-5, IL-13) is measured by ELISA or multiplex assays from cell culture supernatants. T-cell proliferation is assessed by 3H-thymidine incorporation or CFSE dilution following 72 hours of stimulation. IC50 and EC50 values are calculated from dose-response curves using nonlinear regression analysis.
|
| Animal Protocol |
Animal/Disease Models: balb/c (Bagg ALBino) mouse and SD (SD (Sprague-Dawley)) rats[1]
Doses: 1 and 5 mg/kg Route of Administration: Oral (5 mg/kg) and intravenous (iv) (iv)injection (1 mg/kg) Experimental Results: 1.19 Route Doses Volume (mL/kg) Dose (mg/kg) Cmax (μM) AUC (μM·h) Cl (mL/min/kg) T1/2 (h) Vss (L/kg) %F Mouse (BALB/c ) IV 1.0 1.0 1.5 40 2.9 10 Mouse (BALB/c) PO 5.0 50 3.8 2.7 36 Animal/Disease Models: balb/c (Bagg ALBino) mouse and SD (SD (Sprague-Dawley)) rats [1] Doses: 1 and 5 mg/kg Route of Administration: PO (5 mg/kg) and IV (1 mg/kg) Experimental Results: 1.19 Routes Dose Volume (mL/kg) Dose (mg/kg) Cmax (μM) AUC (μM·h) Cl (mL/min/kg ) T1/2 (h) Vss (L/kg) %F Rat (SD (SD (Sprague-Dawley)) IV 1.0) 1.0 0.6 70 3.0 14 Rat (SD (SD (Sprague-Dawley)) PO 2.0 5.0 0.2 1.1 40 In vivo efficacy is studied in murine models of asthma, including ovalbumin (OVA) and house dust mite (HDM)-induced airway inflammation models. GNE-9822 is administered orally at doses of 1-50 mg/kg, typically twice daily (BID) due to its oral bioavailability of 36% in mice. In rats and dogs, the compound is administered at 1-5 mg/kg intravenously or 5-50 mg/kg orally. Airway inflammation is assessed by bronchoalveolar lavage (BAL) fluid cell counts (eosinophils, neutrophils, lymphocytes), histological analysis of lung tissue (H&E and PAS staining for mucus production), and measurement of Th2 cytokine levels (IL-4, IL-5, IL-13) in BAL fluid and lung homogenates by ELISA. Pharmacodynamic markers include ITK phosphorylation status and downstream signaling intermediates in lung tissue. In conscious guinea pigs, GR 125743 (0.3 mg/kg; i.p.) produces significant decreases in extracellular 5-HT in the frontal cortex. |
| ADME/Pharmacokinetics |
GNE-9822 demonstrates good ADME properties with favorable pharmacokinetic profiles across multiple species. In mice, the plasma half-life (t1/2) is 2.9 hours, with an oral bioavailability (F%) of 36%. In rats, the half-life is 3.0 hours. In dogs, the half-life is 5.4 hours with an oral bioavailability of 70%. The compound shows good bioavailability and half-life in mice and rats following oral and intravenous administration at 1 and 5 mg/kg. The oral bioavailability of 36% in mice supports BID dosing in murine OVA/HDM studies. These PK properties enable allometric scaling and support the use of GNE-9822 in preclinical efficacy studies. The compound is soluble in DMSO and should be stored according to standard protocols for small-molecule inhibitors.
|
| Toxicity/Toxicokinetics |
Specific toxicological data for GNE-9822 are not extensively published in publicly available literature. As a research compound, it should be handled with standard laboratory safety precautions, including the use of appropriate personal protective equipment (gloves, lab coat, safety goggles) and adequate ventilation. The compound is for research use only and not for human or veterinary therapeutic applications. Users should consult the material safety data sheet (MSDS) for detailed safety information, including potential hazards, first-aid measures, and disposal considerations. As with all kinase inhibitors, appropriate care should be taken to avoid inhalation, ingestion, or skin contact.
|
| References |
|
| Additional Infomation |
GNE-9822 is a high-quality chemical probe developed for studying ITK biology and T-cell signaling. It is not approved for clinical use and is intended for research purposes only. The compound is classified as a probe by the Chemical Probes Portal with recommended concentrations of IC50: 55 nM and inhibitor up to 100 nM. GNE-9822 was discovered through a property- and structure-guided approach targeting the tetrahydroindazole series of ITK inhibitors. References include the discovery of potent and highly selective ITK degraders with in vivo activity and property- and structure-guided discovery of a tetrahydroindazole series of ITK inhibitors. The compound's exceptional kinome selectivity (>100-fold over ITK Ki against 286 kinases, with only six hits >70% inhibition at 0.1 μM) makes it a valuable tool for validating ITK as a therapeutic target in inflammatory diseases and for dissecting ITK-specific signaling pathways in T-cell biology.
|
| Molecular Formula |
C24H32N6O
|
|---|---|
| Molecular Weight |
420.55
|
| Exact Mass |
420.263
|
| CAS # |
1557232-32-2
|
| PubChem CID |
74983355
|
| Appearance |
Off-white to light yellow solid powder
|
| LogP |
3.8
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
4
|
| Rotatable Bond Count |
7
|
| Heavy Atom Count |
31
|
| Complexity |
605
|
| Defined Atom Stereocenter Count |
1
|
| SMILES |
C1C=CC=CC=1[C@H](CCN(C)C)N1C=C(NC(=O)C2=NNC3CC(C)(CCC2=3)C)C=N1
|
| InChi Key |
XFYUTGIEFKGWND-NRFANRHFSA-N
|
| InChi Code |
InChI=1S/C24H32N6O/c1-24(2)12-10-19-20(14-24)27-28-22(19)23(31)26-18-15-25-30(16-18)21(11-13-29(3)4)17-8-6-5-7-9-17/h5-9,15-16,21H,10-14H2,1-4H3,(H,26,31)(H,27,28)/t21-/m0/s1
|
| Chemical Name |
N-[1-[(1S)-3-(dimethylamino)-1-phenylpropyl]pyrazol-4-yl]-6,6-dimethyl-1,4,5,7-tetrahydroindazole-3-carboxamide
|
| Synonyms |
GNE9822; GNE 9822; GNE-9822
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| Solubility (In Vitro) |
DMSO : ~62.5 mg/mL (~148.61 mM)
|
|---|---|
| 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 | 2.3778 mL | 11.8892 mL | 23.7784 mL | |
| 5 mM | 0.4756 mL | 2.3778 mL | 4.7557 mL | |
| 10 mM | 0.2378 mL | 1.1889 mL | 2.3778 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
