Antimicrobial agent 325 (compound 14a) (0.25-16 μg/mL) exhibits potent broad-spectrum in vitro antimicrobial activity, particularly against methicillin-resistant Staphylococcus aureus (MIC = 0.25 μg/mL), Escherichia coli (MIC = 0.25 μg/mL), Klebsiella pneumoniae (MIC = 0.5 μg/mL), and Acinetobacter baumannii (MIC = 0.5 μg/mL) [1]. Antimicrobial agent 325 (0.5-128 μg/mL) exhibits extremely low hemolytic activity against human erythrocytes, with hemolysis rates below 3% even at concentrations up to 128 μg/mL [1]. Antimicrobial agent 325 (10-100 μg/mL; 24 h) shows low cytotoxicity against LO2 human hepatocytes [1]. Antimicrobial agent 325 and norfloxacin showed synergistic antimicrobial activity against both Escherichia coli and methicillin-resistant Staphylococcus aureus (MRSA) in vitro, with a FICI of 0.25 for both strains [1]. Antimicrobial agent 325 (0.25-1 μg/mL; 12 h) exhibited rapid bactericidal activity against MRSA, completely eliminating the strain within 12 hours at 4 times the MIC concentration [1]. Antimicrobial agent 325 (16 days) had a low probability of inducing MRSA resistance, and no increase in MIC was observed after 16 consecutive passages [1]. Antimicrobial agent 325 (0.25-1 μg/mL; 10-30 min) induced significant, time-dependent membrane depolarization in methicillin-resistant Staphylococcus aureus (MRSA) [1]. Antimicrobial agent 325 (0.25–1.5 μg/mL) disrupts the membrane integrity of MRSA in a dose-dependent manner, manifested as increased phosphatidylinositol (PI) uptake and decreased calcein-AM retention [1]. Antimicrobial agent 325 (0.25–2 μg/mL) can cause dose-dependent protein leakage in MRSA [1]. Antimicrobial agent 325 (0.25–2.5 μg/mL; 30–150 min) can induce dose-dependent and time-dependent reactive oxygen species (ROS) production in MRSA [1]. Antimicrobial agent 325 (0.25–2 μg/mL) can induce dose-dependent lipid peroxidation in MRSA [1]. Antimicrobial agent 325 (0.25–2 μg/mL) impairs the antioxidant defense system of methicillin-resistant Staphylococcus aureus (MRSA) cells, resulting in a dose-dependent decrease in glutathione (GSH) activity [1]. Antimicrobial agent 325 (2 μg/mL; 6 h) caused visible morphological damage and cell shrinkage in MRSA[1]. Antimicrobial agent 325 (0.25–2 μg/mL) inhibited the metabolic activity of MRSA in a dose-dependent manner[1]. Antimicrobial agent 325 (0.25–2 μg/mL) inhibited the lactate dehydrogenase (LDH) activity of MRSA in a dose-dependent manner[1].

Antimicrobial agent 325 (compound 14a) (0.100 mg/kg; topical application; once every 48 hours; twice) can inhibit the inflammatory response caused by bacterial infection and promote wound healing in a mouse model of MRSA infection[1]. Antimicrobial agent 325 (0.067-0.1 mg/kg) can improve the survival rate of MRSA-infected larvae of the large wax moth[1].

Cytotoxicity assay [1]
Cell Types: LO2 human hepatocytes
Tested Concentrations: 10-100 μg/mL
Incubation Duration: 24 hours
Experimental Results: At the bactericidal concentration, cell viability remained above 90%, and even at a concentration of 100 μg/mL, viability remained above 60%.

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Immunofluorescence [1]
Cell Types: MRSA
Tested Concentrations: 0.5 μg/mL
Incubation Duration: 6 hours Experimental
Experimental Results: Enhanced fluorescence of DCFH-DA.

Animal/Disease Models:MRSA-infected ICR mice (wound infection model) [1]
Doses: 0.100 mg/kg
Route of Administration: Topical administration
Experimental Results: Reduced neutrophil levels. At a dose of 0.100 mg/kg, the relative area of the wound in mice decreased to nearly 0% on day 7, comparable to the effect of 0.400 mg/kg ciprofloxacin.

[1]. Discovery of novel structural imidazolylvinylquinolones exerting excellent broad-spectrum antibacterial efficacy with multitargeting potential. Eur J Med Chem. 2026;307:118643.

C25H25CLFN3O3

469.94

Typically exists as solids at room temperature

O=C1C2=CC(F)=C(/C(C#N)=C/C3=C(Cl)C=C(CCCC)N3CC)C=C2N(CC)C=C1C(O)=O

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

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

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.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*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).

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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)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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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).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)