| Size | Price | Stock | Qty |
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| 100mg |
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| Other Sizes |
| Targets |
MurA (UDP-N-acetylglucosamine enolpyruvyl transferase). MurA-IN-4 is a covalent inhibitor of MurA, an essential enzyme that catalyzes the first committed step in peptidoglycan biosynthesis: the transfer of enolpyruvate from phosphoenolpyruvate (PEP) to UDP-N-acetylglucosamine (UNAG). Inhibition of MurA disrupts bacterial cell wall synthesis, leading to bacterial cell death. The chloroacetamide warhead covalently binds to a cysteine residue in the active site.
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| ln Vitro |
MurA-IN-4 exhibits antibacterial activity by inhibiting the bacterial cell wall synthesis enzyme MurA. It inhibits MurA enzyme activity and shows antibacterial effects against bacterial strains, particularly those expressing MurA. While specific IC50 values are not provided, the compound has demonstrated potent inhibition of MurA in biochemical assays and antibacterial effects in cell-based assays.
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| ln Vivo |
No specific in vivo activity data has been reported. As a MurA inhibitor with antibacterial activity, MurA-IN-4 would be evaluated in animal models of bacterial infection, such as murine thigh infection models or peritonitis models. It is expected to reduce bacterial burden in infected tissues.
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| Enzyme Assay |
For cell-free MurA inhibition assay, purified recombinant MurA (e.g., from E. coli) is incubated with increasing concentrations of MurA-IN-4 (0.1 nM-100 uM) in 50 mM HEPES buffer, pH 7.5, containing 1 mM DTT, 5 mM MgCl2, and 0.1% BSA at 25degC for 10-30 minutes. The reaction is initiated by adding UDP-N-acetylglucosamine (UNAG, 50 uM) and phosphoenolpyruvate (PEP, 100 uM). The reaction proceeds for 30-60 minutes at 37degC and is stopped by adding 0.5 M HCl. The product, enolpyruvyl-UNAG (EP-UNAG), is quantified by measuring absorbance at 232 nm using a spectrophotometer or by HPLC. IC50 is determined by plotting inhibition percentage against inhibitor concentration. The chloroacetamide warhead forms a covalent bond with a cysteine residue in the active site, which can be confirmed by mass spectrometry.
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| Cell Assay |
For in vitro antibacterial assays, standard bacterial strains (e.g., E. coli ATCC 25922, S. aureus ATCC 29213, or MurA-expressing strains) are cultured overnight in Mueller-Hinton broth. Cultures are diluted to 0.5 McFarland standard (approximately 10⁶ CFU/mL). MurA-IN-4 is serially diluted (0.1-100 ug/mL) in Mueller-Hinton broth in 96-well plates. Bacteria are added to achieve a final inoculum of 5×10⁵ CFU/mL. Plates are incubated at 37degC for 18-24 hours. Minimum inhibitory concentration (MIC) is determined as the lowest concentration that prevents visible bacterial growth. For time-kill kinetics, cultures treated with MurA-IN-4 at 1×, 2×, and 4× MIC are sampled at 0, 2, 4, 6, 12, and 24 hours, and viable colonies are enumerated by plating. The integrity of the bacterial cell wall can be assessed by measuring leakage of cytoplasmic enzymes (e.g., lactate dehydrogenase) or by electron microscopy.
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| Animal Protocol |
No specific in vivo animal study protocols are documented. A typical protocol for evaluating a MurA inhibitor would involve establishing a murine systemic infection model. Female BALB/c mice (6-8 weeks old) are challenged intraperitoneally with a lethal dose of E. coli or S. aureus (approximately 5×10⁷ CFU/mouse) suspended in 10% mucin. One hour post-infection, MurA-IN-4 is administered via intravenous or intraperitoneal injection at doses ranging from 1-50 mg/kg. A positive control group receives a standard antibiotic (e.g., ciprofloxacin or vancomycin). Survival is monitored for 7-14 days. Alternatively, in a neutropenic murine thigh infection model, mice are rendered neutropenic by cyclophosphamide administration (150 mg/kg, i.p., 4 days and 1 day before infection). Thighs are injected with 10⁵-10⁶ CFU of bacteria. MurA-IN-4 is administered subcutaneously at various doses (0.1-100 mg/kg) 2 hours post-infection. After 24 hours, mice are euthanized, and thigh muscles are homogenized for bacterial colony counting. Efficacy is expressed as reduction in log10 CFU/g compared to untreated controls.
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| ADME/Pharmacokinetics |
No pharmacokinetic data has been reported for MurA-IN-4. As a small molecule with a molecular weight of 205.64 g/mol and molecular formula C8H12ClNO3, the compound may have moderate oral bioavailability. Its ADME (absorption, distribution, metabolism, excretion) properties have not been characterized. The chloroacetamide warhead suggests potential for covalent binding to serum proteins, which may affect PK profile.
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| Toxicity/Toxicokinetics |
No specific toxicity data has been reported. As a MurA inhibitor with a chloroacetamide warhead, the compound is a potential alkylating agent and may exhibit genotoxicity or cytotoxicity at high concentrations. Formal toxicological evaluation (acute, subchronic, genotoxicity) has not been performed. Standard chemical safety precautions (gloves, goggles, fume hood) should be used when handling this compound. It is not recommended for human use.
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| References |
[1]. Grabrijan K, et al. Covalent inhibitors of bacterial peptidoglycan biosynthesis enzyme MurA with chloroacetamide warhead. Eur J Med Chem. 2022 Dec 5;243:114752.
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| Additional Infomation |
MurA-IN-4 is also known as 1-(2-chloroacetyl)piperidine-4-carboxylic acid. The molecular formula is C8H12ClNO3, and the molecular weight is 205.64. The IUPAC name is 1-(2-chloroacetyl)piperidine-4-carboxylic acid. The compound contains a chloroacetamide warhead that covalently binds to a cysteine residue in the active site of MurA, leading to irreversible inhibition of the enzyme. It is a research chemical for antibacterial and anti-infective studies. It is not approved for clinical or veterinary use. It should be stored as a powder at -20degC and is stable for up to 3 years.
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| Molecular Formula |
C8H12CLNO3
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| Molecular Weight |
205.64
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| Exact Mass |
205.051
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| CAS # |
318280-69-2
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| PubChem CID |
1991140
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| Appearance |
Typically exists as solid at room temperature
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| Density |
1.349g/cm3
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| Boiling Point |
406.2ºC at 760 mmHg
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| Melting Point |
109-111ºC
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| Flash Point |
199.4ºC
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| Index of Refraction |
1.526
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| LogP |
0.486
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
13
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| Complexity |
211
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1CN(CCC1C(=O)O)C(=O)CCl
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| InChi Key |
PEQVHRULPWCLOJ-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C8H12ClNO3/c9-5-7(11)10-3-1-6(2-4-10)8(12)13/h6H,1-5H2,(H,12,13)
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| Chemical Name |
1-(2-chloroacetyl)piperidine-4-carboxylic acid
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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 | 4.8629 mL | 24.3143 mL | 48.6287 mL | |
| 5 mM | 0.9726 mL | 4.8629 mL | 9.7257 mL | |
| 10 mM | 0.4863 mL | 2.4314 mL | 4.8629 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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