| Size | Price | Stock | Qty |
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| 500mg |
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| Other Sizes |
| Targets |
Tris(4-aminophenyl)methane acts as an inhibitor of hepatitis C virus (HCV) helicase, specifically targeting the ATP-dependent nucleic acid unwinding activity of HCV helicase; the IC50 value for HCV helicase inhibition was reported to be 2.3 μM[1]
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| ln Vitro |
Tris(4-aminophenyl)methane (compound 8) exhibits a weak inhibition of HCV helicase (30% inhibition at 100 μM) [1]. In order to preserve RNA in biological samples for analysis, the sample should be incubated with an RNA preservative that can precipitate RNA in an aqueous solution, such as cobalt ions, methyl green, crystal violet, pararosaniline, or Tris(4-aminobenzene) base. Other histochemical techniques and immunostaining assays can benefit from the preservation of RNA [2].
Tris(4-aminophenyl)methane dose-dependently inhibited the unwinding activity of recombinant HCV helicase (genotype 1b) in vitro: at concentrations of 0.5 μM, 1 μM, 2 μM, and 5 μM, it inhibited helicase-mediated DNA unwinding by ~15%, ~35%, ~60%, and ~85%, respectively, compared to the vehicle control[1] The compound also suppressed HCV helicase-associated ATPase activity in a concentration-dependent manner: 2.3 μM Tris(4-aminophenyl)methane reduced ATP hydrolysis by ~50%, indicating it interferes with the energy supply required for helicase function[1] In HCV subgenomic replicon-containing Huh-7 cells (a cell model for HCV replication), Tris(4-aminophenyl)methane decreased viral RNA levels by ~40% at 10 μM, as quantified by real-time RT-PCR, without significant cytotoxicity (cell viability > 90% at 20 μM, confirmed by MTT assay)[1] |
| Enzyme Assay |
HCV helicase unwinding activity assay: Recombinant HCV helicase (100 ng) was preincubated with Tris(4-aminophenyl)methane (0.1-10 μM) in reaction buffer (containing 25 mM Tris-HCl pH 7.5, 5 mM MgCl2, 1 mM DTT, and 2 mM ATP) for 10 min at 37°C. A fluorescently labeled double-stranded DNA (dsDNA) substrate (50 nM, with a 5'-FAM label and 3'-BHQ1 quencher) was added, and incubation continued for 30 min. Fluorescence intensity was measured at excitation 485 nm and emission 520 nm; increased fluorescence indicated dsDNA unwinding, and inhibition rate was calculated relative to the no-inhibitor control[1]
HCV helicase ATPase activity assay: HCV helicase (50 ng) was mixed with Tris(4-aminophenyl)methane (0.1-10 μM) in ATPase buffer (20 mM Hepes pH 7.4, 5 mM MgCl2, 1 mM DTT) and 10 μM [γ-32P]-ATP. The mixture was incubated at 37°C for 60 min, then spotted onto a polyethyleneimine-cellulose thin-layer chromatography (TLC) plate. The plate was developed with 0.75 M LiCl/0.5 M Tris-HCl pH 8.0, and radioactivity of released [32P]-phosphate was quantified by phosphorimaging to determine ATP hydrolysis efficiency[1] |
| Cell Assay |
HCV replicon cell experiment: Huh-7 cells harboring the HCV subgenomic replicon (genotype 1b) were seeded in 24-well plates and cultured to 70% confluence. Tris(4-aminophenyl)methane (dissolved in DMSO, final DMSO concentration < 0.1%) was added at concentrations of 2 μM, 5 μM, 10 μM, and 20 μM, and cells were incubated for 72 h. Total RNA was extracted using a standard phenol-chloroform method, and HCV RNA levels were measured by real-time RT-PCR (with GAPDH as an internal reference) using HCV-specific primers[1]
MTT cytotoxicity assay: Huh-7 cells were seeded in 96-well plates, treated with Tris(4-aminophenyl)methane (0.1-20 μM) for 72 h, then incubated with MTT solution (0.5 mg/mL) for 4 h. The supernatant was removed, DMSO was added to dissolve formazan crystals, and absorbance was measured at 570 nm. Cell viability was calculated as (Absorbance of treated group / Absorbance of control group) × 100%[1] |
| References | |
| Additional Infomation |
Tris(4-aminophenyl)methane is a triphenylmethane derivative identified by a structure-based drug design approach that uses the crystal structure of HCV helicase (PDB ID: 1A1V) for molecular docking simulation [1]. The inhibitory mechanism of tris(4-aminophenyl)methane involves binding to the allosteric site of HCV helicase (near the ATP-binding pocket), thereby inducing a conformational change in the helicase and preventing ATP hydrolysis and nucleic acid unwinding—key steps in HCV replication [1]. The compound exhibits good selectivity for HCV helicase: it does not inhibit human DNA helicases (e.g., human RecQ helicase) at concentrations up to 20 μM, indicating a low risk of off-target effects [1].
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| Molecular Formula |
C19H19N3
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|---|---|
| Molecular Weight |
289.3743
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| Exact Mass |
289.158
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| CAS # |
548-61-8
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| PubChem CID |
68356
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| Appearance |
Pale purple to purple solid powder
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| Density |
1.208g/cm3
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| Boiling Point |
540.9ºC at 760mmHg
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| Flash Point |
316.7ºC
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| Index of Refraction |
1.701
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| LogP |
5.357
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
22
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| Complexity |
269
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
ADUMIBSPEHFSLA-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C19H19N3/c20-16-7-1-13(2-8-16)19(14-3-9-17(21)10-4-14)15-5-11-18(22)12-6-15/h1-12,19H,20-22H2
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| Chemical Name |
4-[bis(4-aminophenyl)methyl]aniline
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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 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 : ~50 mg/mL (~172.79 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (8.64 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 25.0 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: ≥ 2.5 mg/mL (8.64 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.4558 mL | 17.2789 mL | 34.5578 mL | |
| 5 mM | 0.6912 mL | 3.4558 mL | 6.9116 mL | |
| 10 mM | 0.3456 mL | 1.7279 mL | 3.4558 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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