| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| Other Sizes |
| Targets |
ML318 targets PvdQ (an Ntn hydrolase involved in pyoverdine synthesis) with an in vitro IC50 of 20 nM. [1]
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| ln Vitro |
ML318 is a biaryl nitrile inhibitor of PvdQ acylase. ML318 inhibits PvdQ in vitro (IC50 = 20 nM) by binding in the acyl-binding site, as confirmed by the X-ray crystal structure of PvdQ bound to ML318. Additionally, the PvdQ inhibitor is active in a whole cell assay, preventing pyoverdine production and limiting the growth of P. aeruginosa under iron-limiting conditions.[1]
ML318 inhibited PvdQ acylase activity with an IC50 of 20 nM in a fluorogenic biochemical assay using 4-methylumbelliferyl laurate as substrate. [1] In whole-cell assays, ML318 reduced pyoverdine production in P. aeruginosa PAO1 with an IC50 of 1.9 μM (determined by direct HPLC measurement of pyoverdine). [1] In 48-hour growth cultures, ML318 exhibited an IC50 of 19 ± 4 μM against PAO1 wild-type (measured by absorbance at 405 nm for pyoverdine production). Against PAK wild-type strain, IC50 was 43 ± 18 μM; against a pump mutant (mexAB-oprM) of PAK, IC50 improved to 1.4 ± 0.2 μM. [1] ML318 showed no activity in other PubChem Bioassays, including toxicity studies with other bacteria (M. tuberculosis and E. coli). [1] |
| Enzyme Assay |
Recombinant PvdQ protein was prepared. The enzymatic substrate 4-methylumbelliferyl (4-MU) laurate was formulated using isopropanol, Triton X-100, and TNT buffer. PvdQ (0.02 μM) and 4-MU laurate (0.8 mM) were mixed in 1536-well plates with compounds added acoustically. Positive control: isopropyl dodecylfluorophosphate (IDFP) at 200 μM. Fluorescence was measured at excitation 303-367 nm and emission 440-460 nm at time 0 and after 60 min at room temperature. The fluorescence difference (60 min minus 0 min) was calculated. IC50 values were determined using nine compound concentrations ranging from 3 nM to 19.5 μM with 20 nM PvdQ. [1]
X-ray crystal structure of PvdQ bound to ML318 (compound 4) was determined (see PDB structure). The compound binds in the acyl-binding site with the nitrile surrounded by Pro401, Trp402, and Val403, and the aromatic rings engaging in π-stacking with Phe240 and Trp378. The trifluoromethyl group on the pyridine interacts with hydrophobic residues Phe240, Ile274, Trp378, Trp402, and Val403. [1] |
| Cell Assay |
Whole-cell pyoverdine production assay: P. aeruginosa (PAO1) was grown for 4 h in the presence of compounds, and the amount of pyoverdine produced was directly measured using HPLC. Pyoverdine migrates as a cluster of peaks that were integrated for IC50 calculation. For ML318, the whole-cell IC50 was 1.9 μM. [1]
Growth inhibition assay: P. aeruginosa strains (PAO1, PAK wild-type, and PAK mexAB-oprM pump mutant) were grown for 48 h in iron-limiting conditions (with EDDHA) in the presence of compound concentrations. Absorbance at 405 nm (pyoverdine production) and 600 nm (growth) were measured. IC50 values were calculated from dose-response curves. [1] Stability assay: ML318 (compound 4) showed improved stability over compound 3; after 48 h in neutral PBS buffer, about 30-40% of compound 3 remained, while compound 4 showed modestly better stability. After 6 h in the presence of 50 μM glutathione, about 90% of compound 3 remained; compound 4 was similarly stable. [1] |
| ADME/Pharmacokinetics |
ML318 (compound 4) was tested for toxicity in a HeLa cell counter screen and showed no toxicity. It also was not active in other PubChem Bioassays, including toxicity studies with M. tuberculosis and E. coli. [1]
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| Toxicity/Toxicokinetics |
ML318 (designated as Probe Compound ML318) is a biaryl nitrile with the structure shown in Figure 5 and Table 6 (R1 = 2-CF3, R2 = H, R3 = F on pyridine; phenyl ring with 4-fluorophenyl). The mechanism of action is competitive inhibition of PvdQ by binding in the fatty acyl-binding pocket. ML318 reduces pyoverdine production and limits growth of P. aeruginosa under iron-limiting conditions. It was shown to be active against two P. aeruginosa strains (PAO1 and PAK) and its activity was enhanced in an efflux pump mutant, indicating that efflux mechanisms partly account for the difference between in vitro and whole-cell potencies. Combination with 5-fluorocytosine (which reduces expression of pyoverdine synthesis genes) showed synergistic effect (IC50 of 1.7 μM for the combination vs 7.9 μM for ML318 alone). No in vivo animal data are reported. [1]
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| References |
| Molecular Formula |
C14H8F4N2
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|---|---|
| Molecular Weight |
280.220336914063
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| Exact Mass |
280.06
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| Elemental Analysis |
C, 60.01; H, 2.88; F, 27.12; N, 10.00
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| CAS # |
1610516-67-0
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| PubChem CID |
56604881
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| Appearance |
Solid powder
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| LogP |
3.3
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
20
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| Complexity |
365
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
DXQNDKQUHKVTTC-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C14H8F4N2/c15-10-6-4-9(5-7-10)11(8-19)12-2-1-3-13(20-12)14(16,17)18/h1-7,11H
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| Chemical Name |
(4-Fluoro-phenyl)-(6-trifluoromethyl-pyridin-2-yl)-acetonitrile
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| Synonyms |
ML318; ML 318; ML-318; CID-56604881
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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 Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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 : ~100 mg/mL (~356.86 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 2.5 mg/mL (8.92 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: 2.5 mg/mL (8.92 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. View More
Solubility in Formulation 3: 10% DMSO+40% PEG300+5% Tween-80+45% Saline: 2.5 mg/mL (8.92 mM) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.5686 mL | 17.8431 mL | 35.6862 mL | |
| 5 mM | 0.7137 mL | 3.5686 mL | 7.1372 mL | |
| 10 mM | 0.3569 mL | 1.7843 mL | 3.5686 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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