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| Other Sizes |
| Targets |
EcDsbB-IN-9 targets EcDsbB, the E. coli disulfide bond formation protein B (DsbB). DsbB is a membrane protein that catalyzes the oxidation of DsbA, which is essential for the formation of disulfide bonds in periplasmic proteins. By inhibiting EcDsbB, the compound disrupts disulfide bond formation, leading to the accumulation of misfolded proteins and bacterial cell death.
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| ln Vitro |
EcDsbB-IN-9 (compound 9) (0-100 μM) suppresses DsbA oxidation with an IC50 of 8.5 ± 0.6 μM and accumulates decreased DsbA (EcDsbB substrate) [1].
In vitro, EcDsbB-IN-9 potently inhibits EcDsbB with an IC50 of 1.7 μM and a Ki of 46 nM. It is active against E. coli with an MIC of 5.7 µM. The compound is a specific EcDsbB inhibitor. These activities confirm its potential as an antimicrobial agent targeting disulfide bond formation in Gram-negative bacteria. |
| ln Vivo |
In vivo, EcDsbB-IN-9 has potential as an antimicrobial agent for treating Gram-negative bacterial infections. By targeting EcDsbB, it disrupts bacterial protein folding and function, leading to bacterial death. However, detailed in vivo efficacy data in animal models of infection is limited in publicly available sources. The compound is primarily used as a research tool.
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| Enzyme Assay |
In vitro enzyme assays for EcDsbB-IN-9 measure inhibition of EcDsbB activity using a coupled assay with DsbA and a substrate such as insulin or a fluorescent peptide. DsbB oxidizes DsbA, which then oxidizes the substrate. Various concentrations of EcDsbB-IN-9 are added. Enzyme activity is measured by monitoring substrate oxidation. IC50 and Ki values are calculated from dose-response curves.
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| Cell Assay |
Cellular assays for EcDsbB-IN-9 use E. coli cultures. Bacteria are treated with various concentrations of the compound. Bacterial growth is measured by optical density at 600 nm. Minimum inhibitory concentration (MIC) is determined by broth microdilution according to CLSI guidelines. Disulfide bond formation in periplasmic proteins may be assessed by SDS-PAGE under non-reducing conditions.
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| Animal Protocol |
In vivo animal studies for EcDsbB-IN-9 would utilize models of Gram-negative bacterial infection, such as murine peritonitis or sepsis models. The compound would be administered intraperitoneally or intravenously. Bacterial load in tissues would be measured. Survival would be monitored. Efficacy would be compared to vehicle and reference antibiotics. However, detailed in vivo data is limited.
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| ADME/Pharmacokinetics |
EcDsbB-IN-9 has molecular weight of 255.1 g/mol and molecular formula C11H8Cl2N2O. It is also known as 4,5-dichloro-2-benzyl-3(2H)-pyridazinone. Purity is typically ≥98%. It is a solid. It is soluble in DMSO. It is typically stored as powder at -20°C for up to 3 years. Pharmacokinetic properties would need to be determined.
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| Toxicity/Toxicokinetics |
Preclinical toxicity of EcDsbB-IN-9 has not been extensively documented. As an antimicrobial agent targeting a bacterial enzyme, potential adverse effects would be evaluated in standard toxicology studies. The compound's selectivity for bacterial DsbB over human proteins is important for its safety profile. No significant toxicity has been reported.
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| References |
[1]. Landeta C, et al. Compounds targeting disulfide bond forming enzyme DsbB of Gram-negative bacteria. Nat Chem Biol. 2015 Apr;11(4):292-8.
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| Additional Infomation |
EcDsbB-IN-9 (Compound 9) is a potent E. coli DsbB inhibitor with an IC50 of 1.7 μM and a Ki of 46 nM. It is active against E. coli with an MIC of 5.7 µM. It targets the disulfide bond-forming enzyme DsbB in Gram-negative bacteria. It is available for research purposes only.
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| Molecular Formula |
C11H8CL2N2O
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|---|---|
| Molecular Weight |
255.098
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| Exact Mass |
254.001
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| CAS # |
41933-33-9
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| PubChem CID |
722438
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| Appearance |
Typically exists as solid at room temperature
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| Density |
1.38g/cm3
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| Boiling Point |
338ºC at 760 mmHg
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| Melting Point |
87ºC
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| Flash Point |
158.2ºC
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| Index of Refraction |
1.626
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| LogP |
2.598
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
2
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
16
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| Complexity |
346
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| Defined Atom Stereocenter Count |
0
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| SMILES |
ClC1=C(C=NN(C1=O)CC1C=CC=CC=1)Cl
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| InChi Key |
AJHBQZQCDCTOFD-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C11H8Cl2N2O/c12-9-6-14-15(11(16)10(9)13)7-8-4-2-1-3-5-8/h1-6H,7H2
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| Chemical Name |
2-benzyl-4,5-dichloropyridazin-3-one
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| Synonyms |
EcDsbB-IN-9; EcDsbB IN 9; EcDsbBIN9
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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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.9200 mL | 19.6002 mL | 39.2003 mL | |
| 5 mM | 0.7840 mL | 3.9200 mL | 7.8401 mL | |
| 10 mM | 0.3920 mL | 1.9600 mL | 3.9200 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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