| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| Other Sizes |
| Targets |
IC50: 0.026 μM (TNIK)[1], 0.030 μM (Flt4)[1], 0.191 μM (Flt1)[1], 0.411 μM (DRAK1)[1]
TNIK (Traf2- and Nck-interacting protein kinase) is the primary target of TNIK-IN-3, with an IC50 of 0.026 μM. The compound also shows inhibitory activity against Flt4 (IC50 = 0.030 μM), Flt1 (IC50 = 0.191 μM), and DRAK1 (IC50 = 0.411 μM). Additionally, it exhibits activity against Aurora-A, GCK, and MLK3 with IC50 values of 0.517 μM, 3.657 μM, and 4.552 μM, respectively. |
|---|---|
| ln Vitro |
Compound 21k, TNIK-IN-3, inhibits GCK, MLK3, and Aurora-A with IC50 values of 3.657 μM, 4.552 μM, and 0.517 μM, respectively[1]. The viability of HCT116 and DLD-1 cells is inhibited by TNIK-IN-3 (0.1-100 μM; 3 days), with IC50s of 4.26 μM and 8.00 μM, respectively[1]. HCT116 and DLD-1 cell colony formation is dose-dependently inhibited by TNIK-IN-3 (2.5–40 μM; 10 days)[1]. TNIK-IN-3 (5–20 μM; 48 h) prevents DLD-1 and HCT116 cell migration[1]. In HCT116 cells, TNIK-IN-3 (5–40 μM; 48 h) dose-dependently suppresses the expression of the Wnt target genes AXIN2 and c-Myc, as well as the LRP5 and LRP6 proteins[1]. In Hela cells, TNIK-IN-3 (5–20 μM; 48 h) dramatically reduces JNK1/2 phosphorylation [1].
In vitro, TNIK-IN-3 (compound 21k) reduces HCT116 and DLD-1 cell viability in a dose-dependent manner, with IC50 values of 4.26 μM and 8.00 μM over three days at concentrations ranging from 0.1 to 100 μM. It inhibits colony formation over 10 days (2.5 to 40 μM) and cell migration over 48 hours (5 to 20 μM). The compound also dose-dependently decreases LRP5 and LRP6 protein expression, as well as the Wnt target genes AXIN2 and c-Myc in HCT116 cells (5 to 40 μM, 48 hours). |
| ln Vivo |
TNIK-IN-3 (compound 21k) suppresses tumor development in a dose-dependent manner (100–150 mg/kg; po twice daily for 18 days)[1].
In vivo data for TNIK-IN-3 are limited. As an orally active TNIK inhibitor, it is expected to be suitable for oral administration in animal models. Based on its mechanism of action targeting Wnt/β-catenin signaling and its in vitro efficacy in colorectal cancer cell lines, the compound is anticipated to demonstrate tumor growth inhibition in xenograft models. Further in vivo studies are needed to fully characterize its pharmacokinetic and pharmacodynamic profile. |
| Enzyme Assay |
The in vitro enzyme/receptor binding (non-cellular) assay typically involves kinase activity assays using recombinant TNIK protein. The compound is incubated with the kinase, ATP, and a peptide substrate, and the inhibitory activity is measured by quantifying the phosphorylation level. IC50 values are determined by fitting dose-response curves. For TNIK-IN-3, the IC50 of 0.026 μM was established using such a biochemical assay. Similar assays are used for off-target profiling against a panel of kinases to assess selectivity.
|
| Cell Assay |
Cell Viability Assay[1]
Cell Types: HCT116 and DLD-1 cells Tested Concentrations: 0.1-100 μM Incubation Duration: 3 days Experimental Results: Inhibited cell viability in a dose-dependent manner. Cell Viability Assay[1] Cell Types: HCT116 cells Tested Concentrations: 5, 10, 20, 40 μM Incubation Duration: 48 hrs (hours) Experimental Results: Inhibited the expression of Wnt target genes AXIN2 and c-Myc, LRP5 and LRP6 proteins. Cellular assays for TNIK-IN-3 are performed using colorectal cancer cell lines such as HCT116 and DLD-1. Cells are treated with varying concentrations of the compound (0.1–100 μM) for 72 hours, and cell viability is measured using standard assays like MTT or CellTiter-Glo to determine IC50 values. Colony formation assays are conducted over 10 days with concentrations of 2.5–40 μM, and cell migration is assessed using wound-healing or transwell assays over 48 hours. |
| Animal Protocol |
Animal/Disease Models: Sixweeks old female NOD-SCID (severe combined immunodeficient) mouse were injected with HCT116 cells[1]
Doses: 100, 150 mg/kg Route of Administration: Po twice (two times) daily for 18 days Experimental Results: Dramatically inhibited tumor growth at a dose of 150 mg/kg. No obvious weight loss and no other side effects were observed. In vivo animal studies for TNIK-IN-3 are expected to utilize mouse xenograft models of colorectal cancer. Tumor-bearing mice would be administered the compound orally at various doses. Tumor volume and body weight would be monitored regularly to assess efficacy and tolerability. Pharmacodynamic markers such as AXIN2 and c-Myc expression in tumor tissues would be analyzed. However, specific in vivo protocols for this compound have not been detailed in the available literature. |
| ADME/Pharmacokinetics |
Detailed pharmacokinetic (PK) properties of TNIK-IN-3 have not been fully characterized in the available literature. However, as an orally active compound, it is expected to have sufficient oral bioavailability and systemic exposure following oral administration. Further studies are required to determine its half-life, clearance, volume of distribution, and metabolic stability. The compound is likely to be formulated in standard vehicles for in vivo studies, such as DMSO, PEG300, and Tween 80.
|
| Toxicity/Toxicokinetics |
Comprehensive toxicity data for TNIK-IN-3 are not available. As a research compound intended for preclinical studies, standard safety pharmacology and toxicology assessments would be required before clinical development. The compound is not approved for human use and is intended for research purposes only. Given its kinase inhibitory profile, potential off-target effects should be carefully evaluated in future studies to assess the risk of toxicity.
|
| References | |
| Additional Infomation |
TNIK-IN-3 is also known as compound 21k and is a specific and orally bioactive TNIK antagonist. It shows potential for research on colorectal cancer (CRC) by modulating the Wnt/β-catenin signaling pathway. The compound is not yet in clinical trials and is currently in the preclinical research stage. Its high potency and selectivity make it a valuable chemical probe for studying TNIK biology and validating TNIK as a therapeutic target.
|
| Molecular Formula |
C23H18FN3O2
|
|---|---|
| Molecular Weight |
387.41
|
| Exact Mass |
387.138
|
| CAS # |
2754265-25-1
|
| PubChem CID |
162641720
|
| Appearance |
Off-white to light yellow solid powder
|
| LogP |
3.7
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
4
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
29
|
| Complexity |
583
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
O1C2=CC(C3=CN=C4NC=CC4=C3)=CC=C2C(=O)N(CC2=CC=C(F)C=C2)CC1
|
| InChi Key |
UVLOISNMRLZTPF-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C23H18FN3O2/c24-19-4-1-15(2-5-19)14-27-9-10-29-21-12-16(3-6-20(21)23(27)28)18-11-17-7-8-25-22(17)26-13-18/h1-8,11-13H,9-10,14H2,(H,25,26)
|
| Chemical Name |
4-[(4-fluorophenyl)methyl]-8-(1H-pyrrolo[2,3-b]pyridin-5-yl)-2,3-dihydro-1,4-benzoxazepin-5-one
|
| 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 : 83.33 mg/mL (215.10 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 | 2.5812 mL | 12.9062 mL | 25.8124 mL | |
| 5 mM | 0.5162 mL | 2.5812 mL | 5.1625 mL | |
| 10 mM | 0.2581 mL | 1.2906 mL | 2.5812 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
