| Targets |
The compound targets dihydrofolate synthetase, an enzyme involved in folate biosynthesis. It also acts as an inhibitor of cysteine proteases, forming a covalent bond with the thiol group of cysteine residues, leading to irreversible enzyme inactivation. It interacts with MurA and other bacterial enzymes, making it a broad-spectrum antimicrobial agent.
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| ln Vitro |
In vitro, 2,4-Dinitrobenzenesulfonamide exhibits significant antibacterial and antifungal activity. It shows inhibitory activity against various microorganisms, including Aspergillus niger, Candida albicans, Bacillus subtilis, and Staphylococcus aureus. It is also a potent MurA inhibitor with an IC50 of 164 uM, and it targets dihydrofolate synthetase, disrupting bacterial folate metabolism and cell growth.
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| ln Vivo |
In vivo, 2,4-Dinitrobenzenesulfonamide is used as an antibacterial and antifungal agent. Its efficacy has been demonstrated in animal models of bacterial infection. However, due to its reactivity and potential toxicity, it is primarily used as a research compound rather than a clinical therapeutic. In vivo studies typically focus on its mechanism of action and its utility as a chemical probe.
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| Enzyme Assay |
For dihydrofolate synthetase assays, the enzyme (0.1-1.0 ug) is incubated with 2,4-Dinitrobenzenesulfonamide (0.1-100 uM) in a buffer (50 mM Tris-HCl, pH 8.0, 5 mM MgCl2, 5 mM ATP) at 37degC for 20-30 minutes. The substrate (dihydropteroate and glutamate) is added, and the reaction is terminated by adding 10% trichloroacetic acid. Product formation is measured by HPLC or by a coupled spectrophotometric assay. IC₅0 values are calculated from dose-response curves.
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| Cell Assay |
For antimicrobial susceptibility testing, bacteria (e.g., S. aureus, B. subtilis) or fungi (e.g., C. albicans, A. niger) are cultured in appropriate media (e.g., Mueller-Hinton broth for bacteria, Sabouraud dextrose broth for fungi). 2,4-Dinitrobenzenesulfonamide is added at concentrations ranging from 0.1-200 ug/mL in 96-well plates. After incubation at 37degC for 18-24 hours (bacteria) or 30degC for 24-48 hours (fungi), the minimum inhibitory concentration (MIC) is determined as the lowest concentration that inhibits visible growth.
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| Animal Protocol |
For in vivo efficacy studies, immunocompromised or normal mice (e.g., BALB/c) are infected with a lethal dose of bacteria (e.g., S. aureus, ~1×10⁸ CFU/mouse) via intraperitoneal injection. Two hours post-infection, 2,4-Dinitrobenzenesulfonamide is administered intraperitoneally or orally at doses of 10-100 mg/kg in a vehicle (e.g., 2% DMSO in saline) twice daily for 3-5 days. Survival is monitored daily, and bacterial load in blood, liver, and spleen is quantified by CFU counting at the endpoint.
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| ADME/Pharmacokinetics |
2,4-Dinitrobenzenesulfonamide is a small, polar molecule (MW 247.19) with limited oral bioavailability due to its low water solubility. It is typically administered intraperitoneally in animal studies. After absorption, it is rapidly metabolized in the liver via reduction of nitro groups and acetylation of the sulfonamide. The elimination half-life is expected to be 1-3 hours in rodents. The compound is excreted primarily in urine as metabolites.
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| Toxicity/Toxicokinetics |
2,4-Dinitrobenzenesulfonamide is an irritant to skin and eyes. It may cause allergic skin reactions. The compound is a potential mutagen due to the presence of nitro groups, which can be reduced to reactive intermediates that damage DNA. Prolonged or repeated exposure may cause organ damage. It should be handled with care in a well-ventilated fume hood, wearing appropriate PPE.
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| References | |
| Additional Infomation |
2,4-Dinitrobenzenesulfonamide is a valuable research tool for studying dihydrofolate synthetase and cysteine proteases. It is used to synthesize sulfonamide-based drug candidates with antibacterial, antifungal, and anticancer properties. The compound serves as a starting material for the synthesis of more complex bioactive molecules. It is not approved for clinical use and is intended for laboratory research only.
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| Exact Mass |
246.99
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|---|---|
| CAS # |
73901-01-6
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| PubChem CID |
10911854
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| Appearance |
White to yellow solid powder
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| Hydrogen Bond Donor Count |
1
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
16
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| Complexity |
390
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
HMMRSEKWXWQVIW-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C6H5N3O6S/c7-16(14,15)6-2-1-4(8(10)11)3-5(6)9(12)13/h1-3H,(H2,7,14,15)
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| Chemical Name |
2,4-dinitrobenzenesulfonamide
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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.) |
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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