| Size | Price | Stock | Qty |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| Other Sizes |
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Purity: ≥98%
TH257 (TH-257; TH 257) is a novel and selective allosteric inhibitor of LIM kinases-LIMK1 and LIMK2 with important biological activity. It can be used as a chemical probe for LIMK1 and LIMK2, as it inhibits them with IC50 values of 39 nM and 84 nM, respectively. An αC and DFG-out conformation-induced binding pocket is the target of the allosteric inhibitor TH-257. TH257 exhibits remarkable selectivity, as demonstrated by the KINOMEscan assay at 1 μM, which shows no discernible activity against the broader kinome.
| Targets |
LIMK1 (IC50 = 84 nM); LIMK2 (IC50 = 39 nM)
- LIM kinase 1 (LIMK1) (IC50: 84 nM) [1, 2] - LIM kinase 2 (LIMK2) (IC50: 39 nM) [1, 2] |
|---|---|
| ln Vitro |
- Enzyme inhibition: TH257 potently inhibits LIMK1/2 activity in biochemical assays, with IC50 values of 84 nM (LIMK1) and 39 nM (LIMK2). It acts as an allosteric inhibitor targeting a binding pocket induced by αC-helix and DFG-out conformations [1, 2].
- Cofilin phosphorylation inhibition: In cell-based assays, TH257 blocked LIMK-mediated cofilin phosphorylation, a key step in actin cytoskeleton regulation, with submicromolar potency [2]. - Kinome selectivity: At 1 μM, TH257 showed no significant activity against 468 other kinases in kinome-wide screening, confirming its high selectivity for LIMK1/2 [1, 2]. Promising biochemical and cellular effectiveness are demonstrated by TH-257. However, it is not a feasible candidate for use as an in vivo instrument due to its exceedingly fast exterior clearance [2]. |
| Enzyme Assay |
- LIMK1/2 kinase activity assay: Recombinant LIMK1/2 enzymes were incubated with ATP and a fluorescent peptide substrate. TH257 was titrated into the reaction mixture, and phosphorylation was measured via fluorescence resonance energy transfer (FRET). IC50 values were determined by nonlinear regression analysis [1, 2].
- Allosteric binding mode analysis: Surface plasmon resonance (SPR) and molecular modeling studies revealed that TH257 binds to an allosteric pocket distinct from the ATP-binding site, stabilizing the inactive DFG-out conformation of LIMK1/2 [2]. |
| Cell Assay |
- Cofilin phosphorylation assay: Human cancer cells (e.g., MDA-MB-231) were treated with TH257 (0.1–10 μM) for 2 hours. Phosphorylated cofilin levels were quantified by Western blot using phospho-specific antibodies, showing dose-dependent inhibition [2].
- Neurite outgrowth assay: In N1E-115 neuroblastoma cells, TH257 (1–10 μM) dose-dependently suppressed neurite extension, consistent with its role in actin dynamics regulation [2]. |
| ADME/Pharmacokinetics |
- In vitro clearance: TH257 exhibited extremely rapid clearance in liver microsome assays, rendering it unsuitable for in vivo studies [1, 2].
- Solubility: Soluble in DMSO (250 mg/mL) and formulated in 10% DMSO/40% PEG300/5% Tween-80/45% saline for in vivo administration [1]. |
| References |
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| Additional Infomation |
- Mechanism of action: TH257 disrupts actin cytoskeleton remodeling by inhibiting LIMK1/2-mediated cofilin phosphorylation, potentially blocking cancer cell migration and invasion [1, 2].
- Chemical probe status: Developed as a tool compound for studying LIMK1/2 function, TH257 is widely used in cell biology research but has not advanced to clinical trials due to poor pharmacokinetic properties [1, 2]. |
| Molecular Formula |
C24H26N2O3S
|
|---|---|
| Molecular Weight |
422.5398
|
| Exact Mass |
422.166
|
| Elemental Analysis |
C, 68.22; H, 6.20; N, 6.63; O, 11.36; S, 7.59
|
| CAS # |
2244678-29-1
|
| Related CAS # |
2244678-29-1
|
| PubChem CID |
137796294
|
| Appearance |
Light yellow to yellow solid powder
|
| LogP |
4.7
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
4
|
| Rotatable Bond Count |
9
|
| Heavy Atom Count |
30
|
| Complexity |
611
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
S(C1C=CC(=CC=1)C(N(CC1C=CC=CC=1)CCCC)=O)(NC1C=CC=CC=1)(=O)=O
|
| InChi Key |
VNCIWNGCMAKKEO-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C24H26N2O3S/c1-2-3-18-26(19-20-10-6-4-7-11-20)24(27)21-14-16-23(17-15-21)30(28,29)25-22-12-8-5-9-13-22/h4-17,25H,2-3,18-19H2,1H3
|
| Chemical Name |
N-benzyl-N-butyl-4-(phenylsulfamoyl)benzamide
|
| Synonyms |
TH 257; TH257; 2244678-29-1; N-Benzyl-N-butyl-4-(N-phenylsulfamoyl)benzamide; N-benzyl-N-butyl-4-(phenylsulfamoyl)benzamide; CHEMBL4790618; C24H26N2O3S; TH-257
|
| 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: 85~250 mg/mL (201.2~591.7 mM)
Ethanol: ~85 mg/mL (~201.2 mM) |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.92 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 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. Solubility in Formulation 2: ≥ 2.08 mg/mL (4.92 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 20.8 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. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (4.92 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3666 mL | 11.8332 mL | 23.6664 mL | |
| 5 mM | 0.4733 mL | 2.3666 mL | 4.7333 mL | |
| 10 mM | 0.2367 mL | 1.1833 mL | 2.3666 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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