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| 10mg |
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| Targets |
The biological target of AHL modulator-1 is the LuxR-type family of transcriptional regulators, which are the intracellular receptors for AHL signals in many Gram-negative pathogens. Specifically, it modulates the activity of QS receptors such as AbaR in Acinetobacter baumannii and QscR in Pseudomonas aeruginosa, acting as both a partial agonist and an antagonist. It is a phenylacetanoyl L-homoserine lactone (PHL) derivative that strongly modulates many LuxR-type receptors. AHL modulator-1 does not target traditional drug targets like enzymes or GPCRs; rather, it interferes with bacterial quorum sensing by binding to these receptors, thereby preventing them from responding to natural AHL signals. The consequence is the disruption of coordinated bacterial behaviors without affecting bacterial growth, thus exerting less evolutionary pressure for resistance development.
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| ln Vitro |
The in vitro activity of AHL modulator-1 has been quantitatively characterized using two major assays: cellulase activity and potato maceration. In cellulase activity assays (a measure of a key virulence factor in some phytopathogens), AHL modulator-1 exhibits both agonistic (21%) and antagonistic (42%) properties at the same concentration. In potato maceration assays (a measure of tissue degradation and bacterial virulence on plant hosts), it shows agonistic (5%) and antagonistic (32%) activities, respectively. This mixed profile indicates that the compound is a partial modulator, capable of both activating and inhibiting QS-regulated functions depending on the specific receptor and cellular context. Its activity is typically measured in bacterial cultures without significant growth inhibition, confirming that its effects are due to QS modulation rather than general toxicity.
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| ln Vivo |
In vivo activity of AHL modulator-1 was evaluated using a plant infection model (potato maceration). In this model, the compound's ability to attenuate the virulence of plant-pathogenic bacteria (such as Pectobacterium carotovorum or similar species) was assessed by measuring the reduction in tissue maceration (soft rot) volume. The results showed that AHL modulator-1 exhibited both agonistic (5%) and antagonistic (32%) activity in the potato maceration assay, indicating its potential to both slightly promote and more significantly reduce bacterial virulence in a living host system. While the compound shows promise as an anti-virulence agent, no studies have been reported in mammalian animal models (e.g., mice or rats) for systemic infections. Its utility has thus far been demonstrated in plant pathosystems, which are valuable for studying the fundamentals of quorum sensing modulation in vivo.
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| Enzyme Assay |
There is no standard protocol for direct enzyme/receptor binding for AHL modulator-1 in a cell-free system, as it is not an enzyme inhibitor but a receptor modulator. A standard protocol for studying its interaction with LuxR-type receptors involves a reporter gene assay using a bacterial biosensor strain. (1) Culture a biosensor strain (e.g., E. coli carrying a plasmid with a QS receptor like LuxR and a promoter-lacZ or GFP fusion) overnight. (2) Dilute the culture 1:100 into fresh media. (3) Add a fixed concentration of a natural AHL signal (e.g., 3-oxo-C6-HSL) along with varying concentrations of AHL modulator-1 (0.1-100 uM) to the wells of a microtiter plate. (4) Incubate at 30degC for 4-6 hours with shaking to allow for gene expression. (5) For beta-galactosidase assays, lyse the cells, add the substrate (e.g., ONPG or CPRG), and measure absorbance at 420 nm (ONPG) or 570 nm (CPRG). For GFP reporters, measure fluorescence (Ex/Em = 485/528 nm). (6) To determine antagonist activity, co-incubate with a sub-saturating concentration of the natural agonist (AHL) and measure the reduction in signal. To determine agonist activity, add the modulator alone in the absence of natural AHL and measure the induction of the reporter.
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| Cell Assay |
An in vitro protocol for assessing AHL modulator-1 uses a cellulase activity assay in a bacterial culture to measure its effect on a secreted virulence factor. (1) Grow a virulent bacterial strain (e.g., Pectobacterium carotovorum or Dickeya dadantii) overnight at 28degC in liquid culture. (2) Dilute the overnight culture to an OD600 of 0.1 in fresh culture medium containing the cell density inducer (e.g., autoinducer). (3) Aliquot the bacterial suspension into sterile culture tubes or flasks. (4) Add AHL modulator-1 (dissolved in DMSO) to the cultures at a range of concentrations (e.g., 10, 50, 100, 200 uM) and a vehicle control (DMSO only). (5) Incubate at 28degC for 18-24 hours with shaking. (6) Centrifuge the cultures to pellet bacteria. (7) Collect the supernatant and assay for total extracellular cellulase activity using a standard substrate, such as 0.5% carboxymethylcellulose (CMC) in buffer, incubated at 50degC for 30-60 minutes. (8) Measure the reducing sugars released using a method such as the dinitrosalicylic acid (DNS) assay by reading absorbance at 540 nm. (9) Calculate the % modulation (agonism or antagonism) compared to untreated control.
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| Animal Protocol |
AHL modulator-1 was evaluated in an in vivo plant infection model known as the potato maceration assay. A typical protocol: (1) Prepare a bacterial suspension of a virulent, quorum-sensing-dependent strain (e.g., Pectobacterium carotovorum subsp. carotovorum) in sterile phosphate-buffered saline (PBS) to a final concentration of ~10⁸ CFU/mL. (2) AHL modulator-1 is dissolved in an appropriate solvent (e.g., DMSO) and diluted in the bacterial suspension to a final concentration of 100-500 uM (with a final DMSO concentration ≤ 1%). (3) Sterile, fresh potatoes are sliced into uniform cylinders or disks (approx. 1 cm thick) using a cork borer. (4) The potato disks are surface-sterilized by dipping in 70% ethanol and air-drying. (5) A small well is bored into the center of each potato disk. (6) The bacterial suspension, with or without the test compound, is inoculated into the well (e.g., 20 uL). (7) The inoculated potato disks are placed in sterile Petri dishes containing moist filter paper to maintain humidity. (8) After incubation at 28degC for 48-72 hours, the extent of tissue maceration (soft rot) is quantified by measuring the volume (mm3) of decayed tissue using a ruler, or by weighing the macerated tissue. (9) The percent activity (agonism vs. antagonism) is calculated by comparing maceration volume in compound-treated samples to positive (bacteria only, 100% maceration) and negative (PBS only, 0% maceration) controls.
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| ADME/Pharmacokinetics |
AHL modulator-1 is a research chemical, and detailed pharmacokinetic (PK) studies have not been published. The molecular weight is 298.13. For in vitro experiments, it is soluble in DMSO at concentrations up to 125 mg/mL (419.28 mM). For in vivo (plant) studies, it can be dissolved in organic solvents like DMSO or ethanol and then diluted in aqueous buffers, ensuring the final organic solvent concentration is low (≤1%) to avoid phyto-toxicity. Storage: The powder should be stored at -20degC, protected from light and moisture. It is stable for up to 3 years as a powder. Stock solutions in DMSO should be stored at -80degC for up to 6 months or at -20degC for up to 1 month, and repeated freeze-thaw cycles should be avoided. No data are available regarding its absorption, distribution, metabolism, and excretion (ADME) in animals or humans. LogP, pKa, and plasma protein binding data are not available.
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| Toxicity/Toxicokinetics |
There are no published toxicity data for AHL modulator-1 in mammalian systems, as it is a research compound primarily studied in plant and bacterial models. It is not a pharmaceutical and has not been evaluated in standard preclinical toxicology studies. The compound contains a brominated aromatic ring, which raises the potential for metabolic activation to reactive intermediates. However, the concentrations used in bacterial and plant studies (e.g., 100-200 uM) did not exhibit general growth inhibition or significant phytotoxicity, suggesting a degree of selectivity for quorum sensing machinery over general cellular metabolism. Standard safety precautions should be applied when handling this compound: wear appropriate personal protective equipment (PPE), including gloves, safety glasses, and a lab coat. Work should be performed in a well-ventilated area (fume hood). Avoid skin contact, inhalation of dust/aerosols, and ingestion. It is strictly for research use and not approved for human or veterinary use.
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| References | |
| Additional Infomation |
AHL modulator-1 (compound 12) is a phenylacetanoyl L-homoserine lactone (PHL) used to study attenuation of bacterial virulence by targeting quorum sensing (QS) pathways. PHLs are structural analogs of natural AHL autoinducers. The compound was originally described in a study by Palmer et al. (ACS Chemical Biology, 2011) which showed that synthetic QS modulators could function under native conditions on plant hosts, reducing the need for traditional antibiotics. The advantage of anti-virulence (or quorum quenching) strategies is that they disarm pathogens without killing them, theoretically reducing the selective pressure that drives antibiotic resistance. The compound exhibits a mixed agonism/antagonism profile: in cellulase activity assays, it shows 21% agonism and 42% antagonism; in potato maceration assays, it shows 5% agonism and 32% antagonism. No clinical trials or regulatory approvals exist for this compound.
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| Molecular Formula |
C12H12BRNO3
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| Molecular Weight |
298.13
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| Exact Mass |
297
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| CAS # |
942296-18-6
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| PubChem CID |
25014621
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| Appearance |
Solid powder ; White to off-white
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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 |
3
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| Heavy Atom Count |
17
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| Complexity |
308
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| Defined Atom Stereocenter Count |
1
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| SMILES |
C1COC(=O)[C@H]1NC(=O)CC2=CC=CC=C2Br
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| InChi Key |
MMJSTOMMZPSKSQ-JTQLQIEISA-N
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| InChi Code |
InChI=1S/C12H12BrNO3/c13-9-4-2-1-3-8(9)7-11(15)14-10-5-6-17-12(10)16/h1-4,10H,5-7H2,(H,14,15)/t10-/m0/s1
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| Chemical Name |
2-(2-bromophenyl)-N-[(3S)-2-oxooxolan-3-yl]acetamide
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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) |
DMSO : ~125 mg/mL (~419.28 mM; with ultrasonication)
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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 | 3.3542 mL | 16.7712 mL | 33.5424 mL | |
| 5 mM | 0.6708 mL | 3.3542 mL | 6.7085 mL | |
| 10 mM | 0.3354 mL | 1.6771 mL | 3.3542 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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