| Size | Price | Stock | Qty |
|---|---|---|---|
| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| Other Sizes |
| Targets |
SIRT1 (IC50 = 17 μM)
SIRT2 (IC50 = 74 μM) SIRT3 (IC50 = 235 μM) [1] |
|---|---|
| ln Vitro |
In HCT116 cells, SIRT-IN-3 (Compound 7) (30-300 μM; 8 hours) acetates the p53 gene[1].
2-(phenylamino)benzamide inhibited SIRT1 with an IC50 of 17 μM and showed 4-fold selectivity over SIRT2 (IC50 = 74 μM) and 14-fold selectivity over SIRT3 (IC50 = 235 μM). [1] It did not inhibit class I and class II HDACs at a concentration of 1000 μM. [1] Enzyme kinetic analysis revealed that compound 7 acts as a noncompetitive inhibitor with respect to NAD+ and a competitive inhibitor with respect to the acetylated lysine substrate. [1] In cellular assays, compound 7 (20, 50, 100 μM) increased the acetylation level of p53 in HCT116 cells after etoposide-induced DNA damage in a dose-dependent manner. [1] |
| Enzyme Assay |
SIRT1 inhibition assay: Initial screening was performed at 300 μM compound concentration. For IC50 determination, serial dilutions of 2-(phenylamino)benzamide were used to measure deacetylation of a fluorogenic substrate. [1]
Enzyme kinetic assay: To investigate the inhibition mechanism, various concentrations of compound 7 (0, 50, 150, 300 μM) were incubated with SIRT1, NAD+, and acetylated lysine substrate. Reaction rates were measured, and Lineweaver-Burk plots were generated by plotting reciprocal velocity against reciprocal NAD+ concentration (fixed acetylated lysine substrate) and reciprocal acetylated lysine substrate concentration (fixed NAD+). The patterns showed noncompetitive inhibition with NAD+ and competitive inhibition with acetylated lysine substrate. [1] Isoform selectivity assay: SIRT2 and SIRT3 inhibition activities were measured similarly, giving IC50 values of 74 μM and 235 μM, respectively. [1] Class I/II HDAC inhibition assay: Compound 7 was tested at 1000 μM and showed no inhibition. [1] |
| Cell Assay |
HCT116 cells were incubated with 20 μM etoposide to induce DNA damage and subsequent p53 expression, along with various concentrations of 2-(phenylamino)benzamide (20, 50, 100 μM) for 8 hours. Cell lysates were prepared and subjected to SDS-PAGE, followed by western blotting with an antibody specific for acetylated p53. The results showed a dose-dependent increase in acetylation level of p53. [1]
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| References | |
| Additional Infomation |
2-(phenylamino)benzamide was identified from a nicotinamide- and benzamide-focused chemical library. It is structurally similar to EX-527 but exhibits different SAR. Molecular docking into yeast HST2 (PDB code 1Q1A) revealed that the NH2 and CO groups form hydrogen bonds with backbone carbonyl of Ala227 and backbone amine of Tyr229, while the phenyl group interacts with hydrophobic residues (Val182, Phe184, Gly185, Leu188, Val228, Pro238) blocking the entrance of histone H4-binding pocket. An intramolecular hydrogen bond between CO and NH stabilizes the active conformation. [1]
The compound does not inhibit class I/II HDACs at 1000 μM, indicating selectivity for class III HDACs. [1] It increased p53 acetylation in HCT116 cells after etoposide-induced DNA damage, demonstrating cellular SIRT1 inhibition. [1] |
| Molecular Formula |
C13H12N2O
|
|---|---|
| Molecular Weight |
212.25
|
| Exact Mass |
212.095
|
| CAS # |
1211-19-4
|
| PubChem CID |
96455
|
| Appearance |
White to yellow solid powder
|
| Density |
1.218g/cm3
|
| Boiling Point |
413.3ºC at 760 mmHg
|
| Flash Point |
203.8ºC
|
| Vapour Pressure |
4.84E-07mmHg at 25°C
|
| Index of Refraction |
1.663
|
| LogP |
3.302
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
2
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
16
|
| Complexity |
236
|
| Defined Atom Stereocenter Count |
0
|
| InChi Key |
JKWQUMKCVDUICQ-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C13H12N2O/c14-13(16)11-8-4-5-9-12(11)15-10-6-2-1-3-7-10/h1-9,15H,(H2,14,16)
|
| Chemical Name |
2-anilinobenzamide
|
| 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 Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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 : 250 mg/mL (1177.86 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (9.80 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 (9.80 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 (9.80 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 | 4.7114 mL | 23.5571 mL | 47.1143 mL | |
| 5 mM | 0.9423 mL | 4.7114 mL | 9.4229 mL | |
| 10 mM | 0.4711 mL | 2.3557 mL | 4.7114 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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