| Size | Price | Stock | Qty |
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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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| Targets |
Dafadine-A targets DAF-9 (a cytochrome P450) and also inhibits the mammalian functional ortholog CYP27A1. [1]
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| ln Vitro |
Dafadine-A (at 25 µM) induces a protruding vulva (Pvl) phenotype and distal-tip cell migration (Mig) defects in over 60% of wild-type worms.
In a HEK293-cell-based assay, dafadine-A at 20 µM reduced the production of 26-hydroxy-4-cholesten-3-one and Δ4-dafachronic acid by a factor greater than 20 (P < 0.05) in DAF-9-expressing cells. In wild-type worms treated with dafadine-A, reduced amounts of dafachronic acid were measured by LC/MS/MS compared to DMSO-treated controls. Dafadine-A treatment (at a concentration not specified in the main text, but used in L4 larval stage) extended the lifespan of wild-type worms by as much as 29% (P < 0.001). Dafadine-A did not increase the lifespan of daf-9 null mutants, consistent with its action through DAF-9 inhibition. Exogenously supplied dafachronic acid rescued the Daf-c and Mig phenotypes induced by dafadine-A. Dafadine-A inhibits the ability of mammalian CYP27A1 to metabolize 4-cholesten-3-one. Dafadine-A does not inhibit CYP7A1 or CYP7B1. [1] |
| ln Vivo |
Dafadine-A was tested for its ability to antagonize the recovery of daf-9-null mutants from the dauer state. Approximately 75% of daf-9(dh6) null mutants recovered from dauer over an 11-day period, and this recovery was not antagonized by dafadine-A (P > 0.05). [1]
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| Enzyme Assay |
The enzymatic activity of DAF-9 was assessed using a HEK293-cell-based assay. DAF-9 was expressed in HEK293 cells, and the cells were treated with 20 µM 4-cholesten-3-one as substrate. The production of metabolites (26-hydroxy-4-cholesten-3-one and Δ4-dafachronic acid) was measured by liquid chromatography and tandem mass spectrometry (LC/MS/MS). The addition of 20 µM dafadine-A reduced the production of these metabolites by a factor greater than 20 (P < 0.05) in DAF-9(+) cells. Metabolites were normalized for extraction efficiency relative to an internal standard (0.2% EtOH).
Absorbance spectroscopy was used to investigate physical interaction of dafadine-A with DAF-9. Dafadine-A (final concentration 5 µM) was added to Sf9-derived microsomes containing either a human oxidoreductase (HOR)-DAF-9 fusion protein or HOR control protein. A characteristic type 2 difference spectrum was observed with HOR-DAF-9 and dafadine-A, showing an absorption minimum at 407 nm and an absorption maximum at 435 nm, indicating binding to the heme iron. [1] |
| Cell Assay |
Wild-type C. elegans were raised in nonstressful, favorable environmental conditions on media containing increasing concentrations of dafadine-A (0 to 25 µM) to assess Daf-c and Mig phenotypes. For each concentration, four replicates were analyzed with at least 20 animals per replicate.
For epistatic analysis, null alleles of daf-16(m26), daf-12(m20), and din-1(dh127) were used. Worms were treated with dafadine-A (concentration not specified, but consistent with other experiments) and the fraction of worms with Daf-c or Mig phenotype was scored. At least three replicates and at least 90 animals per replicate were analyzed. For lifespan extension studies, worms were treated with dafadine-A at the fourth larval stage (L4). Lifespan was monitored, and dafadine-A treatment extended wild-type lifespan by up to 29% (P < 0.001). No details on solvent formulation, route, or exact concentration are provided in the main text. [1] |
| References | |
| Additional Infomation |
Dafida A is an N-acylpiperidine compound formed by the condensation of the carboxyl group of 5-[(2,6-dimethylphenoxy)methyl]-1,2-oxazol-3-carboxylic acid with the secondary amino group of 4-(pyridin-4-yl)piperidine. It is a P450 inhibitor and an anti-aging drug. It belongs to the pyridine, N-acylpiperidine, isoxazole, aromatic ether, and aromatic amide classes of compounds.
Dafadine-A is the first small-molecule tool that robustly induces dauer formation in Caenorhabditis elegans under typical culture conditions. The Daf-c and Mig phenotypes induced by dafadine-A are completely suppressed by null mutations in daf-12 or din-1S but not by mutations in daf-16, indicating that dafadine-A acts downstream or parallel to DAF-16 but upstream or at the same level as DAF-12. Dafadine-A phenocopies the residual pharyngeal pumping phenotype and SDS sensitivity of daf-9 and daf-12 mutants. Dafadine-A contains an isoxazole substructure and binds to the Fe3+ in the heme pocket of DAF-9, displacing a water molecule required for catalytic activity, as evidenced by a type 2 spectrum in absorbance spectroscopy. Out of 59 inhibitors of mammalian cytochrome P450s tested, none induced Daf-c or Mig phenotype in wild-type worms at 12.5 µM, suggesting that dafadine-A is unique among CYP inhibitors in robustly inhibiting DAF-9 in wild-type worms. Dafadine-A was part of a structure-activity relationship study where 158 structurally related molecules were tested at 12.5 µM for ability to induce Daf-c and/or Mig phenotypes. Only three other molecules (dafadine-B, -C, -D) induced similar phenotypes in wild-type and daf-2(e1370) mutants. [1] |
| Molecular Formula |
C23H25N3O3
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|---|---|
| Molecular Weight |
391.46290564537
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| Exact Mass |
391.189
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| Elemental Analysis |
C, 70.57; H, 6.44; N, 10.73; O, 12.26
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| CAS # |
1065506-69-5
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| PubChem CID |
24790866
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| Appearance |
Light yellow to yellow solid powder
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| Density |
1.2±0.1 g/cm3
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| Boiling Point |
600.3±55.0 °C at 760 mmHg
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| Flash Point |
316.9±31.5 °C
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| Vapour Pressure |
0.0±1.7 mmHg at 25°C
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| Index of Refraction |
1.592
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| LogP |
1.84
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
29
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| Complexity |
522
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O=C(N1CCC(C2=CC=NC=C2)CC1)C3=NOC(COC4=C(C)C=CC=C4C)=C3
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| InChi Key |
WNSNPYIHDMIFOO-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C23H25N3O3/c1-16-4-3-5-17(2)22(16)28-15-20-14-21(25-29-20)23(27)26-12-8-19(9-13-26)18-6-10-24-11-7-18/h3-7,10-11,14,19H,8-9,12-13,15H2,1-2H3
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| Chemical Name |
[5-[(2,6-dimethylphenoxy)methyl]-1,2-oxazol-3-yl]-(4-pyridin-4-ylpiperidin-1-yl)methanone
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| Synonyms |
Dafadine-A
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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 : ~50 mg/mL (~127.73 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.39 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
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 (6.39 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. 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: ≥ 2.5 mg/mL (6.39 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.5545 mL | 12.7727 mL | 25.5454 mL | |
| 5 mM | 0.5109 mL | 2.5545 mL | 5.1091 mL | |
| 10 mM | 0.2555 mL | 1.2773 mL | 2.5545 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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