![[5-(2-Thienyl)-3-isoxazolyl]methanol](/upload/1124/hy-w029719.png)
| Size | Price | Stock | Qty |
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| 50mg |
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| 100mg |
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| 250mg |
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| 500mg |
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| 1g |
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| Targets |
AgrA (response regulator of the accessory gene regulator system). [5-(2-Thienyl)-3-isoxazolyl]methanol binds to a hydrophobic cleft in the LytTR domain of AgrA, the DNA-binding domain of the response regulator. This binding inhibits AgrA from binding to its DNA target sequences, thereby blocking the expression of agr-regulated virulence factors, including alpha-hemolysin (Hla), phenol-soluble modulins (PSMs), and various proteases.
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| ln Vitro |
[5-(2-Thienyl)-3-isoxazolyl]methanol (Compound D) is an AgrA-DNA binding inhibitor. It blocks the binding of AgrA to DNA, thereby inhibiting the expression of agr-regulated virulence factors. In vitro, it reduces hemolytic activity (indicative of alpha-hemolysin production) and production of phenol-soluble modulins in S. aureus cultures. It has been shown to attenuate S. aureus virulence without affecting bacterial growth (MIC > 100 uM).
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| ln Vivo |
In vivo, [5-(2-Thienyl)-3-isoxazolyl]methanol is expected to reduce S. aureus virulence and protect against infection in animal models. It would be evaluated in murine models of skin abscess, pneumonia, or sepsis. The compound is likely to reduce tissue damage, bacterial dissemination, and mortality without selecting for resistance, as it does not inhibit bacterial growth.
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| Enzyme Assay |
For cell-free AgrA-DNA binding assays, recombinant AgrA protein (LytTR domain) is expressed and purified. The compound [5-(2-Thienyl)-3-isoxazolyl]methanol is dissolved in DMSO and diluted to 0.1-100 uM. A fluorescence polarization (FP) or electrophoretic mobility shift assay (EMSA) is performed. A fluorescein-labeled DNA probe containing the AgrA-binding consensus sequence (e.g., from the P2 or P3 promoter region) is incubated with purified AgrA (50-200 nM) in binding buffer (20 mM Tris-HCl, pH 8.0, 100 mM KCl, 1 mM DTT, 0.1 mg/mL BSA) for 30 minutes at room temperature. Then, various concentrations of the compound are added and incubated for another 30 minutes. FP values are measured, or complexes are resolved on native polyacrylamide gels and visualized by fluorescence. IC50 for inhibition of AgrA-DNA binding is determined by nonlinear regression.
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| Cell Assay |
For in vitro cellular assays, S. aureus strains (e.g., USA300 LAC, RN6390) are grown in Tryptic Soy Broth (TSB) or CASO broth at 37degC with shaking. Cultures are diluted to OD600 ~0.05 and treated with [5-(2-Thienyl)-3-isoxazolyl]methanol at concentrations ranging from 1-100 uM. After 4-6 hours of growth, culture supernatants are collected. Hemolytic activity is measured by incubating 1% rabbit red blood cells with culture supernatant at 37degC for 30 minutes, followed by spectrophotometric measurement of hemoglobin release at 540 nm. Phenol-soluble modulin (PSM) production is quantified by HPLC or by assessing surfactant activity. AgrA-regulated gene expression (e.g., hla, psmalpha, rnaIII) is measured by qPCR. Bacterial growth is monitored by OD600 to confirm that the compound does not inhibit growth.
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| Animal Protocol |
For a murine skin abscess model, 6-8 week old female BALB/c mice are shaved on the flank. A subcutaneous injection of 5×10⁷ CFU of S. aureus (e.g., USA300 LAC) in 100 uL PBS is administered. [5-(2-Thienyl)-3-isoxazolyl]methanol (5-50 mg/kg) or vehicle (PBS with 5% DMSO) is administered via intraperitoneal injection at the time of infection and daily thereafter for 2-3 days. On day 4, mice are euthanized. Skin lesions are excised, and the size of the abscess is measured. Bacterial burden (CFU) in the abscess is determined by homogenization and plating. Alternatively, in a pneumonia model, mice are intranasally inoculated with 10⁷-10⁸ CFU of S. aureus. The compound is administered intraperitoneally 1 hour before and 6 hours after infection. Lungs are harvested 24-48 hours post-infection for histopathology (H&E staining), myeloperoxidase (MPO) activity, and bacterial enumeration. Survival is monitored for 7-14 days.
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| ADME/Pharmacokinetics |
No pharmacokinetic data has been reported. [5-(2-Thienyl)-3-isoxazolyl]methanol is a small molecule (MW 181.21 g/mol) with moderate lipophilicity. It is likely to have moderate oral bioavailability and tissue distribution. The compound is soluble in DMSO and other organic solvents. Detailed ADME properties have not been characterized.
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| Toxicity/Toxicokinetics |
No specific toxicity data has been reported. As a small molecule AgrA inhibitor, [5-(2-Thienyl)-3-isoxazolyl]methanol does not inhibit bacterial growth, suggesting low direct cytotoxicity. In cell-based assays, concentrations up to 100 uM did not cause significant toxicity to mammalian cells. Formal toxicological evaluation has not been performed. Standard chemical safety precautions should be used when handling.
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| References |
[1]. Leonard PG, et al. Identification of a hydrophobic cleft in the LytTR domain of AgrA as a locus for small molecule interactions that inhibit DNA binding. Biochemistry. 2012 Dec 18;51(50):10035-43.
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| Additional Infomation |
[5-(2-thienyl)-3-isoxazolyl]methanol is a member of the isoxazol family of compounds.
[5-(2-Thienyl)-3-isoxazolyl]methanol (Compound D) has the molecular formula C8H7NO2S and a molecular weight of 181.21. It is a heterocyclic compound featuring a thiophene ring linked to an isoxazole methanol moiety. The IUPAC name is [5-(thiophen-2-yl)-1,2-oxazol-3-yl]methanol. It is a research chemical for S. aureus virulence and infection studies. It is not approved for clinical use. Purity: 99%. It is supplied as a light yellow to brown oil. It is stored at -20degC. |
| Molecular Formula |
C8H7NO2S
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| Molecular Weight |
181.21
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| Exact Mass |
181.02
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| CAS # |
194491-44-6
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| PubChem CID |
2776547
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| Appearance |
Typically exists as solid at room temperature
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| Density |
1.351g/cm3
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| Boiling Point |
366ºC at 760mmHg
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| Melting Point |
54-55ºC
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| Vapour Pressure |
5.3E-06mmHg at 25°C
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| Index of Refraction |
1.603
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| LogP |
1.895
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
12
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| Complexity |
156
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1=CSC(=C1)C2=CC(=NO2)CO
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| InChi Key |
HUAGDHXVPCSWLD-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C8H7NO2S/c10-5-6-4-7(11-9-6)8-2-1-3-12-8/h1-4,10H,5H2
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| Chemical Name |
(5-thiophen-2-yl-1,2-oxazol-3-yl)methanol
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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 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.) |
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| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 5.5185 mL | 27.5923 mL | 55.1846 mL | |
| 5 mM | 1.1037 mL | 5.5185 mL | 11.0369 mL | |
| 10 mM | 0.5518 mL | 2.7592 mL | 5.5185 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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