| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
TrkA 22.4 nM (IC50)
TrkA-IN-3 specifically targets the TrkA receptor, a receptor tyrosine kinase within the neuronal signaling pathway [21L4-L5]. As a potent and subselective inhibitor, it binds to TrkA with extremely high specificity, blocking its activation by NGF. This inhibition disrupts downstream signaling cascades crucial for pain perception, neuronal survival, and differentiation, positioning TrkA-IN-3 as a key research tool for conditions involving aberrant TrkA activity, such as chronic pain and certain cancers. |
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| ln Vitro |
Compound 5 (TrkA-IN-3) showed 73.9% and 64.8% inhibitory rates against TrkA at 1 μM and 0.1 μM, respectively [1].
In in vitro kinase inhibition assays, TrkA-IN-3 (compound 5) demonstrates 73.9% and 64.8% inhibition of TrkA activity at concentrations of 1 microM and 0.1 microM, respectively [22L19-L21]. Its potency is confirmed by its low IC50 value of 22.4 nM against TrkA, and it has been shown to be more than 8000-fold selective for TrkA over the closely related family members TrkB and TrkC [22L15-L18]. This high selectivity minimizes off-target effects, making it a highly specific tool for studying TrkA biology. |
| ln Vivo |
In vivo efficacy data for TrkA-IN-3 is derived from its use in pain research. While a prodrug (TrkA-IN-4) is often used for in vivo administration, the active form, TrkA-IN-3, is responsible for the potent antinociceptive effects observed. The compound and its related prodrug have been shown to be effective in animal models of pain, validating that selective inhibition of TrkA is a viable strategy for pain relief. The IC50 of 22.4 nM indicates the high potency required for observable effects in vivo.
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| Enzyme Assay |
To perform a cell-free binding assay for TrkA-IN-3, a TR-FRET or ELISA-based kinase assay is commonly used. The assay typically involves incubating a purified, active recombinant TrkA enzyme with a biotinylated peptide substrate in a reaction buffer (e.g., 50 mM HEPES (pH 7.5), 10 mM MgCl2, 1 mM EGTA, 0.01% Brij-35). TrkA-IN-3 is added at a range of concentrations along with 10 microM ATP. After incubation, the reaction is stopped with EDTA. The phosphorylated substrate is detected using a donor (e.g., europium-labeled anti-phosphotyrosine antibody) and an acceptor (e.g., streptavidin-APC) to calculate the IC50 via the TR-FRET signal.
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| Cell Assay |
A standard cell-based assay to assess the activity of TrkA-IN-3 involves treating neuroblastoma or TrkA-expressing cancer cells (e.g., SH-SY5Y, IMR-32, or Ba/F3-TrkA cells). Cells are plated in 96-well plates and treated with a dilution series of the compound (e.g., 0.01-10000 nM) for a specific duration (typically 72 hours). Cell viability is then measured using a luminescent cell viability assay (e.g., CellTiter-Glo 2.0). Inhibition of TrkA autophosphorylation can also be directly measured by Western blotting after a shorter incubation (e.g., 2 hours) with the compound, followed by stimulation with NGF for 10 minutes.
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| Animal Protocol |
In vivo studies of TrkA-IN-3 are commonly conducted using its prodrug, TrkA-IN-4, in rodent pain models. A typical protocol for pain assessment is the hot plate test. Male KM mice (20-24 g) are administered TrkA-IN-4 orally at doses ranging from 0.9375 to 120 mg/kg. At 3 hours post-administration, each mouse is placed on a hot plate (55 +/- 0.5degC), and the latency for the mouse to lick its hind paw or jump is recorded with a cutoff time of 30 seconds to prevent tissue damage. The median effective dose (ED50) for the antinociceptive effect of the active TrkA-IN-3 is calculated from these data [19L25-L29].
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| ADME/Pharmacokinetics |
The pharmacokinetics of the active moiety, TrkA-IN-3, are best characterized via its prodrug, TrkA-IN-4. The prodrug is administered orally, after which it is rapidly absorbed and metabolized to release the active TrkA-IN-3. The pharmacokinetic parameters can be determined by dosing rodents orally, collecting blood at various time points (0, 0.25, 0.5, 1, 2, 4, 8, and 24 hours), and using an LC-MS/MS method to quantify both the remaining prodrug and the generated TrkA-IN-3 in plasma. The resulting plasma concentration-time curve is used to calculate parameters such as Cmax, Tmax, half-life, and AUC.
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| Toxicity/Toxicokinetics |
Formal toxicity data for TrkA-IN-3 is not publicly available. As a research chemical, standard handling precautions should be observed to avoid exposure. In the context of a drug discovery program, the safety profile of a TrkA-IN-3 would be assessed through a series of standard in vitro and in vivo toxicology assays. This would include an Ames test for genotoxicity, a hERG assay to assess potential for cardiac arrhythmia, and repeated-dose toxicity studies in two species (rodent and non-rodent) following oral administration, with monitoring for clinical signs, body weight, and histopathology.
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| References | |
| Additional Infomation |
TrkA-IN-3, with CAS number 3026111-73-6, is a high-quality research tool with high selectivity for TrkA (>8000-fold over TrkB and TrkC). It is a selective small molecule inhibitor with an IC50 of 22.4 nM, and it is used in preclinical studies to explore the role of TrkA in neuronal survival, differentiation, and disease-related pathways. The compound blocks the tyrosine kinase activity of the NGF receptor, effectively disrupting its downstream signaling pathways. It is strictly intended for laboratory research use and is not approved for human use or clinical trials. It is commonly stored at -20degC as a powder [22L6].
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| Molecular Formula |
C24H17F3N4O3
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|---|---|
| Molecular Weight |
466.41
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| Exact Mass |
466.125
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| CAS # |
3026111-73-6
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| PubChem CID |
166176996
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
34
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| Complexity |
731
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CN1C=CC(=N1)C2=C(C=CC(=C2)C(=O)NC3=C(N=CC(=C3)C(=O)O)C4=CC=CC=C4)C(F)(F)F
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| InChi Key |
MLKKBTKDOPHWET-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C24H17F3N4O3/c1-31-10-9-19(30-31)17-11-15(7-8-18(17)24(25,26)27)22(32)29-20-12-16(23(33)34)13-28-21(20)14-5-3-2-4-6-14/h2-13H,1H3,(H,29,32)(H,33,34)
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| Chemical Name |
5-[[3-(1-methylpyrazol-3-yl)-4-(trifluoromethyl)benzoyl]amino]-6-phenylpyridine-3-carboxylic acid
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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 :~50 mg/mL (~107.20 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.36 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 25.0 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 | 2.1440 mL | 10.7202 mL | 21.4404 mL | |
| 5 mM | 0.4288 mL | 2.1440 mL | 4.2881 mL | |
| 10 mM | 0.2144 mL | 1.0720 mL | 2.1440 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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