| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| 100mg | |||
| Other Sizes |
| Targets |
MT-4 targets the TG2/FN complex (Tissue Transglutaminase 2/Fibronectin complex), with a binding affinity to TG2 of Kd = 5.1 μM [1]
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| ln Vitro |
Ovarian cancer (OC) cells become more susceptible to conventional anticancer medications when pretreated with MT4 [1].
- Cell adhesion inhibition: MT-4 inhibited adhesion of ovarian cancer cell lines (SKOV3, OVCAR433, OVCAR5) to fibronectin-coated surfaces in a dose-dependent manner (concentrations ranging from 2 μM to 25 μM), and complete inhibition of cell adhesion was achieved at 25 μM [1] - Cytoskeleton and morphology effects: MT-4 induced membrane ruffling in ovarian cancer cells, delayed the formation of stable focal contacts, disrupted actin cytoskeleton organization, and reduced the number of vinculin-positive focal adhesion points [1] - Signal transduction suppression: MT-4 attenuated the activation of focal adhesion kinase (FAK) and extracellular signal-regulated kinase (ERK) in ovarian cancer cells following cell adhesion to fibronectin, thereby inhibiting "outside-in" signaling mediated by FAK [1] - Chemosensitization: Pretreatment of ovarian cancer cells with 1 μM MT-4 for 72 hours enhanced their sensitivity to paclitaxel; combined treatment with 5 nM paclitaxel reduced cancer cell colony formation by 50% compared to paclitaxel alone [1] |
| ln Vivo |
- Intraperitoneal dissemination inhibition: In NSG mice intraperitoneally injected with SKOV3-GFP ovarian cancer cells, intraperitoneal administration of MT-4 at a dose of 10 μg/kg significantly reduced the adhesion of tumor cells to the peritoneal wall; the number of attached GFP-positive cancer cells in the treatment group was 60% lower than that in the vehicle control group [1]
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| Enzyme Assay |
- Bio-layer Interferometry (BLI) binding assay: The 45 kDa gelatin-binding domain of fibronectin (FN45) was immobilized on streptavidin-coated sensors. Tissue Transglutaminase 2 (TG2) at a concentration of 1 μM was pre-incubated with MT-4 at gradient concentrations (0 μM to 100 μM). The sensor was then immersed in the TG2-MT-4 mixture to measure binding kinetics, and the results showed that MT-4 bound to TG2 with a Kd value of 5.1 μM and a ΔH value of -20.5 kcal/mol [1]
- Isothermal Titration Calorimetry (ITC): MT-4 was titrated into a solution containing TG2, and thermodynamic parameters of the binding interaction were measured. The results confirmed the binding affinity of MT-4 to TG2, with a Kd of 5.1 μM and a ΔH of -20.5 kcal/mol [1] |
| Cell Assay |
- Cell adhesion assay: 96-well plates were coated with fibronectin at a concentration of 5 μg/mL. Ovarian cancer cells (SKOV3, OVCAR433, OVCAR5) were pre-treated with MT-4 at concentrations of 2 μM, 5 μM, 10 μM, and 25 μM for 72 hours. The pre-treated cells were seeded onto the fibronectin-coated plates and incubated for 1 hour. Non-adherent cells were removed by gentle washing, and adherent cells were fixed, stained with crystal violet, and quantified by measuring absorbance [1]
- Focal adhesion and actin cytoskeleton analysis: OVCAR5 cells were pre-treated with 1 μM MT-4 for 72 hours, then seeded onto fibronectin-coated chamber slides and incubated for 30 minutes. The cells were fixed, and immunofluorescent staining was performed to detect vinculin (a marker for focal adhesions), phosphorylated FAK (pFAK), and actin. The stained cells were observed and analyzed using confocal microscopy [1] - Western blot analysis: OVCAR cells were pre-treated with 5 μM MT-4 for 72 hours, then plated on fibronectin-coated surfaces and incubated for 15 minutes, 30 minutes, or 45 minutes. Cell lysates were prepared, and the expression levels of pFAK, total FAK, pERK, and total ERK were detected by Western blot. GAPDH was used as a loading control to ensure equal protein loading [1] |
| Animal Protocol |
- Intraperitoneal dissemination model: 6-8 week-old NSG mice were used for the experiment. SKOV3-GFP ovarian cancer cells (1 × 10^6 cells per mouse) were suspended in phosphate-buffered saline (PBS). MT-4 was dissolved in a mixture of DMSO and PBS at a volume ratio of 1:9 to prepare the administration solution. Mice were randomly divided into two groups: the control group received intraperitoneal injection of SKOV3-GFP cells plus an equal volume of the DMSO/PBS vehicle; the treatment group received intraperitoneal injection of SKOV3-GFP cells plus MT-4 at a dose of 10 μg/kg. Two hours after injection, the peritoneal cavity of each mouse was washed with PBS. Non-adherent cells in the wash solution were recovered and counted using flow cytometry, and the peritoneal wall was examined to quantify the number of attached GFP-positive cancer cells [1]
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| References |
[1]. Sima LE, et al. Small Molecules Target the Interaction between Tissue Transglutaminase and Fibronectin. Mol Cancer Ther. 2019 Jun;18(6):1057-1068.
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| Molecular Formula |
C21H23N5O
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|---|---|
| Molecular Weight |
361.440223932266
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| Exact Mass |
361.19
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| CAS # |
2327925-35-7
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| PubChem CID |
121032298
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| Appearance |
Typically exists as solid at room temperature
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| Density |
1.2±0.1 g/cm3
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| Index of Refraction |
1.681
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| LogP |
2.48
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
27
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| Complexity |
458
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O=C(CC1C=CC=CC=1)NC1C=CC(=CC=1)NC1=NC(C)=CC(=N1)N(C)C
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| InChi Key |
SKUUNAQXXGDUMF-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C21H23N5O/c1-15-13-19(26(2)3)25-21(22-15)24-18-11-9-17(10-12-18)23-20(27)14-16-7-5-4-6-8-16/h4-13H,14H2,1-3H3,(H,23,27)(H,22,24,25)
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| Chemical Name |
N-[4-[[4-(dimethylamino)-6-methylpyrimidin-2-yl]amino]phenyl]-2-phenylacetamide
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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 : ~100 mg/mL (~276.67 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.92 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (6.92 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 | 2.7667 mL | 13.8336 mL | 27.6671 mL | |
| 5 mM | 0.5533 mL | 2.7667 mL | 5.5334 mL | |
| 10 mM | 0.2767 mL | 1.3834 mL | 2.7667 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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