| Size | Price | |
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| 5mg | ||
| 10mg | ||
| 50mg | ||
| 100mg | ||
| Other Sizes |
| Targets |
PKM2 (pyruvate kinase M2) – binds to PKM2 with a binding free energy of -8.9 kcal/mol (molecular docking); activates PKM2 and increases its pyruvate kinase activity in a dose-dependent manner (0-100 μM) [1]
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| ln Vitro |
In Vitro: TEPC466 (10 μM) showed a high degree of selectivity for PKM2 protein over other analytes including free amino acids (His, Arg, Ala, Tyr, Cys, Gly), common ions (Na+, K+, Cl-), peptides (GSH), glucose, and proteins (COX-2, proteinase K, NQO1). When mixed with PKM1, TEPC466 emitted slight fluorescence, but the intensity induced by PKM2 was about 5.8-fold higher than that induced by PKM1. [1]
TEPC466 detected PKM2 with high sensitivity, showing a linear fluorescence intensity increase at 390 nm with PKM2 concentrations from 0 to 20 μg/mL (R² = 0.9884). The detection limit was determined to be 21.25 ng/mL. [1] TEPC466 exhibited an aggregation-induced emission (AIE) property. In a THF/H₂O system, TEPC466 emitted weak fluorescence in water; upon addition of THF (up to 90%), fluorescence intensity increased and the emission peak blue-shifted from 420 to 390 nm. In pure THF, fluorescence intensity decreased significantly along with a blue shift. [1] TEPC466 (0-100 μM) increased the PK activity of recombinant PKM2 in a dose-dependent manner, showing a stronger agonistic effect than TEPP-46 (100 μM). [1] Molecular docking revealed that TEPC466 binds to similar sites on PKM2 as TEPP-46, with the coumarine fragment forming additional interactions with PRO517 in the PKM2 subunit. The binding free energies were -8.9 kcal/mol for TEPC466 and -5.6 kcal/mol for TEPP-46, indicating tighter binding of TEPC466 to PKM2. [1] |
| Enzyme Assay |
Enzyme Assay: PK activity assay: Recombinant PKM2 was incubated with different concentrations of TEPC466 (0-100 μM) or TEPP-46 (100 μM). The pyruvate kinase activity was then measured. TEPC466 significantly increased the PK activity of PKM2 in a dose-dependent manner. [1]
Molecular docking: The crystal structure of PKM2 (PDB number: 4B2D1) was used. The position of the co-crystallized ligand was set as the binding site. Autodock Vina was employed as the docking engine. Gasteiger charge was used for electrostatic interaction calculations. The exhaustiveness parameter was set to 8. The docking space was set to 20 × 20 × 20 ų. All ligands (TEPC466 and TEPP-46) were docked 100 times for sufficient sampling. [1] Selectivity assay: The probe TEPC466 (10 μM) was incubated with free amino acids (His, Arg, Ala, Tyr, Cys, Gly), common ions (Na+, K+, Cl-), peptides (GSH), glucose, and proteins (PKM2, COX-2, proteinase K, NQO1) at 37°C for 30 min. Fluorescence emission spectrum was recorded from 330 to 500 nm at an excitation wavelength of 310 nm with excitation and emission slit widths of 3 nm and 5 nm, respectively. [1] Sensitivity assay: PKM2 protein was diluted in a series of concentrations and incubated with TEPC466 (10 μM) at 37°C for 20 min. Fluorescence emission spectrum was measured from 330 to 500 nm at an excitation wavelength of 310 nm. Detection limit was calculated as 3σb/k, where σb is the standard deviation of blank measurements and k is the slope of fluorescence intensities versus sample concentrations. [1] |
| Cell Assay |
Cell Assay: Cytotoxicity evaluation: HT-29 and HCT 116 cells were incubated with different concentrations of TEPC466 (0-20 μM) for 24 h, then cell viability was detected using CCK-8 kit. When incubated with TEPC466 (0-20 μM), cell viability of both cell lines was above 90%, indicating low toxicity and good biocompatibility. [1]
Bioimaging in living cells: HT-29 and HCT 116 colorectal cancer cells were treated with TEPC466 (20 μM) and incubated for 2 h before fluorescence imaging on a laser confocal microscope (λex: 405 nm). TEPC466 successfully emitted blue fluorescence in both normal cancer cells. PKM2-downregulated cells (treated with si-PKM2) incubated with TEPC466 barely exhibited a fluorescence signal. [1] Western blot: PKM2 protein was validated using PKM2 antibody. Endogenous PKM2 protein was significantly depleted by si-PKM2 compared with the control group. [1] PKM2 concentration detection in cells: PKM2 levels in tumor cells (HT-29, Caco-2, HCT 116) ranged from 290-906 ng/mL, which was significantly higher than that in normal cells (293T and IEC-6, 136-195 ng/mL). [1] |
| ADME/Pharmacokinetics |
The probe showed good solubility in aqueous solutions at concentrations below 10 μM; at 50 μM, it showed proper solubility in water but began to precipitate in THF solution. [1]
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| Toxicity/Toxicokinetics |
TEPC466 showed low toxicity in cell viability assays. When HT-29 and HCT 116 cells were incubated with TEPC466 at concentrations from 0 to 20 μM for 24 h, cell viability remained above 90% for both cell lines, indicating good biocompatibility. [1]
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| References | |
| Additional Infomation |
TEPC466 is a novel TEPP-46-based aggregation-induced emission (AIE) fluorescent probe for the detection of PKM2. It was designed by conjugating coumarin-3-carboxylic acid to TEPP-46 via EDC chemistry. The synthetic yield was about 25.7%. The melting range is 85-87°C. The molecular formula is C₂₇H₂₁N₄O₅S₂, with [M+H]+ calcd 545.0953 and found 545.1297 by HRMS. [1]
TEPC466 can be used as both an indicator and an activator of PKM2. It is the first reported fluorescent probe for PKM2. The probe shows potential for cancer diagnosis and therapy. [1] |
| Molecular Formula |
C27H20N4O5S2
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|---|---|
| Molecular Weight |
544.60
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| Exact Mass |
544.087
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| CAS # |
2621957-88-6
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| PubChem CID |
163196493
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| Appearance |
Off-white to yellow solid powder
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| LogP |
3.1
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
38
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| Complexity |
1080
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C12N(C)C3C=C(S(=O)C)SC=3C=1C=NN(CC1=CC(NC(=O)C3C(=O)OC4=CC=CC=C4C=3)=CC=C1)C2=O
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| InChi Key |
UGBKJKATORQJGR-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C27H20N4O5S2/c1-30-20-12-22(38(2)35)37-24(20)19-13-28-31(26(33)23(19)30)14-15-6-5-8-17(10-15)29-25(32)18-11-16-7-3-4-9-21(16)36-27(18)34/h3-13H,14H2,1-2H3,(H,29,32)
|
| Chemical Name |
N-[3-[(7-methyl-4-methylsulfinyl-9-oxo-3-thia-7,10,11-triazatricyclo[6.4.0.02,6]dodeca-1(8),2(6),4,11-tetraen-10-yl)methyl]phenyl]-2-oxochromene-3-carboxamide
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| Synonyms |
TEPC 466; TEPC-466; 2621957-88-6; TEPC466;
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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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| 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 | 1.8362 mL | 9.1811 mL | 18.3621 mL | |
| 5 mM | 0.3672 mL | 1.8362 mL | 3.6724 mL | |
| 10 mM | 0.1836 mL | 0.9181 mL | 1.8362 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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