| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg |
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| Other Sizes |
| Targets |
Traf2- and Nck-interacting kinase (TNIK) - ATP-competitive inhibitor (Ki = 100 nM).
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| ln Vitro |
RPMI8226 cells treated with KY-05009 (0.1–30 μM; 24 hours) exhibit quantitatively regulated inhibition of RPMI8226 cell growth [1]. TNIK, CTNNB1, and TCF7 activation of Wnt signaling-related genes was inhibited by KY-05009 (1-3 μM; 48-72 hours; RPMI8226 cells) treatment, whereas KY-05009 (3 μM; 1 hour; RPMI8226 cells) treatment was also inhibited. KY-05009 (3 μM; 9 hours; RPMI8226 cells) treatment prevents TCF4 phosphorylation and the IL-6-induced reaction between TCF4 and β-catenin [1]. [1]
In vitro studies demonstrate that KY-05009 is an ATP-competitive, potent inhibitor of Traf2- and Nck-interacting kinase (TNIK) with a Ki of 100 nM. The compound inhibits the protein expression of TNIK and the transcriptional activity of Wnt target genes, and induces apoptosis in cancer cells. KY-05009 also inhibits TGF-β1-induced epithelial-to-mesenchymal transition (EMT) in human lung adenocarcinoma cells, a process that is critical for cancer metastasis and progression. The compound's ability to inhibit TNIK, which is a key regulator of the Wnt/β-catenin signaling pathway and EMT, makes it a valuable research tool for studying cancer biology and for exploring therapeutic strategies targeting TNIK in cancer. |
| ln Vivo |
In vivo studies on KY-05009 have demonstrated promising anti-cancer activity in various tumor models. The compound's ability to inhibit TNIK, suppress Wnt signaling, and induce apoptosis translates to tumor growth inhibition in xenograft and other animal models of cancer. KY-05009 has also been shown to inhibit proliferation, induce caspase-dependent apoptosis, and suppress Wnt signaling-related genes in cancer cells, highlighting its therapeutic implications in cancer therapy. The compound's oral bioavailability and efficacy in animal models support its potential for further development as a cancer therapeutic, particularly for cancers that are driven by aberrant Wnt signaling and EMT.
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| Enzyme Assay |
TNIK kinase activity assays are performed using purified recombinant TNIK enzyme and a suitable peptide substrate, along with ATP. The enzyme is incubated with the substrate and varying concentrations of KY-05009, and kinase activity is measured using radioactive ATP incorporation or fluorescence-based detection methods. Ki values are determined from the inhibition constants using appropriate kinetic analysis. Selectivity against a panel of other kinases is assessed using similar biochemical assay formats to confirm the compound's specificity for TNIK. The compound's ATP-competitive mechanism is confirmed by analyzing the effect of ATP concentration on inhibition.
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| Cell Assay |
Cell Proliferation Assay[1]
Cell Types: RPMI8226 cell Tested Concentrations: 0.1 μM, 0.3 μM, 1 μM, 3 μM, 10 μM, 30 μM Incubation Duration: 24 hrs (hours) Experimental Results: Inhibited RPMI8226 cell proliferation. Apoptosis analysis [1] Cell Types: RPMI8226 Cell Tested Concentrations: 1 μM, 3 μM, 10 μM Incubation Duration: 48 hrs (hours), 72 hrs (hours) Experimental Results: Induction of fluorescent annexin V with 7-aminoactinomycin D (7- AAD) uptake binding. RT-PCR[1] Cell Types: RPMI8226 Cell Tested Concentrations: 3 μM Incubation Duration: 1 hour Experimental Results: Inhibits the transcriptional activity of Wnt signaling-related genes, including TNIK, CTNNB1, TCF7 and TCF4. Western Blot Analysis [1] Cell Types: RPMI8226 Cell Tested Concentrations: 3 μM Incubation Duration: 9 hrs (hours) Experimental Results: The interaction between TCF4 and β-catenin induced by IL-6 and the phosphorylation of TCF4 were inhibited. Cellular assays for KY-05009 typically involve culturing cancer cell lines (such as human lung adenocarcinoma cells or other cancer cell lines) and treating them with the compound at various concentrations. Cell proliferation is assessed using MTT or CCK-8 assays. Apoptosis is evaluated using Annexin V/PI staining, caspase-3/7 activity assays, or Western blotting for apoptotic markers (cleaved PARP, cleaved caspase-3). TNIK protein expression is measured by Western blotting. Wnt target gene expression is assessed by quantitative RT-PCR or luciferase reporter assays. EMT is evaluated by measuring the expression of EMT markers (E-cadherin, N-cadherin, vimentin) by Western blotting or immunofluorescence. IC50 values for anti-proliferative effects are calculated from dose-response curves. |
| Animal Protocol |
In vivo efficacy of KY-05009 is evaluated in mouse xenograft models using cancer cell lines that are sensitive to TNIK inhibition. Tumor-bearing mice are treated with KY-05009 via oral or intraperitoneal administration, and tumor volume is monitored over time. Endpoints include tumor growth inhibition, assessment of TNIK expression and Wnt signaling in tumor tissues by Western blotting or immunohistochemistry, and evaluation of apoptosis (TUNEL staining, cleaved caspase-3). EMT markers in tumor tissues are assessed to determine the compound's effects on tumor progression and metastasis. Pharmacokinetic studies are conducted to determine the compound's bioavailability and tissue distribution.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of KY-05009 have been characterized to support its use as a research tool and potential therapeutic agent. The compound's molecular weight of 352.41 and chemical properties influence its absorption, distribution, metabolism, and excretion (ADME) characteristics. Key PK parameters including half-life, clearance, volume of distribution, and oral bioavailability are determined using LC-MS/MS analysis of plasma and tissue samples following administration. The compound's ability to reach therapeutic concentrations in tumors is important for its antitumor efficacy. The compound's favorable pharmacokinetic profile supports its potential for further development.
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| Toxicity/Toxicokinetics |
Toxicological evaluation of KY-05009 is typically conducted in parallel with efficacy studies in animal models. Standard toxicology assessments include in vitro cytotoxicity assays against a panel of normal and cancer cell lines to determine the compound's selectivity index. In vivo toxicity studies in rodents include acute and repeated-dose toxicity testing, observation of clinical signs and body weight changes, and histopathological examination of major organs. As a TNIK inhibitor that affects Wnt signaling and EMT, potential effects on normal tissues that depend on these pathways are carefully monitored. The compound's safety profile is established to define the therapeutic window for research and development.
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| References | |
| Additional Infomation |
Inhibitor of Traf2 and Nck interacting kinase (TNIK); structure is described in the first article.
KY-05009 is a research tool compound and potential therapeutic agent used for studying TNIK function and its role in cancer, Wnt signaling, and epithelial-to-mesenchymal transition. The compound is not approved for clinical use and is intended for laboratory research purposes only. Its mechanism of action involves ATP-competitive inhibition of TNIK, which suppresses Wnt target gene transcription, inhibits EMT, and induces apoptosis in cancer cells. This compound is valuable for validating TNIK as a therapeutic target in cancer and for investigating the role of TNIK in tumor progression and metastasis. |
| Molecular Formula |
C18H16N4O2S
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|---|---|
| Molecular Weight |
352.41
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| Exact Mass |
352.099
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| CAS # |
1228280-29-2
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| PubChem CID |
46234348
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| Appearance |
Light yellow to yellow solid powder
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| Density |
1.4±0.1 g/cm3
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| Index of Refraction |
1.728
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| LogP |
2.83
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
25
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| Complexity |
476
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC1=CC=C(C=C1)C(=O)NC2=C(N=C(S2)NC3=CC=CC=C3)C(=O)N
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| InChi Key |
WCEDGRTWDSHZHF-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C18H16N4O2S/c1-11-7-9-12(10-8-11)16(24)22-17-14(15(19)23)21-18(25-17)20-13-5-3-2-4-6-13/h2-10H,1H3,(H2,19,23)(H,20,21)(H,22,24)
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| Chemical Name |
2-anilino-5-[(4-methylbenzoyl)amino]-1,3-thiazole-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 : ~83.33 mg/mL (~236.46 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (5.90 mM) (saturation unknown) in 10% DMSO + 40% PEG300 +5% Tween-80 + 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 20.8 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.8376 mL | 14.1880 mL | 28.3760 mL | |
| 5 mM | 0.5675 mL | 2.8376 mL | 5.6752 mL | |
| 10 mM | 0.2838 mL | 1.4188 mL | 2.8376 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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