| Size | Price | Stock | Qty |
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| 25mg |
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| 500mg |
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| 1g |
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Purity: ≥98%
Daphnetin (NSC-633563; NSC633563; 7,8-dihydroxycoumarin), a naturally occuring coumarin analog isolated from plants of the genus Daphne, is a PKI-protein kinase inhibitor with important biological activity (e.g. anti-inflammatory and anti-oxidant effects). The IC50 values of 7.67 μM, 9.33 μM, and 25.01 μM, respectively, indicate its inhibition of multikinases, including EGFR, PKA, and PKC. The tyrosine phosphorylation of exogenous substrate by the EGF receptor is strongly inhibited by daphnetin, as is the activity of PKA and PKC. A 24-hour exposure to daphnetin results in the inhibition of the estrogen-responsive human carcinoma cell line MCF-7, with an IC50 of 73 μM. Even at 50 μM of daphnetin, cyclin D1 levels are decreased..
| Targets |
Plasmodium; EGFR ( IC50 = 7.67 μM ); PKA ( IC50 = 9.33 μM ); PKC ( IC50 = 25.01 μM )
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| ln Vitro |
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| ln Vivo |
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| Enzyme Assay |
PKA and PKC activity is found. In short, for the PKC assay, 5 mL of 40 mM cAMP in 50 mM Tris-HCl, pH 7.5, and 5 mL of daphnetin are combined with 5 mL of lipid preparation that contains 100 mM phorbol 12-myristate 13-acetate, 2.8 mg/mL phosphatidyl serine, and Triton X-100 mixed micelles. Ten milliliters of either the PKC substrate solution (10 mL) or the PKA substrate solution (10 mL)—which contains 200 mM Kemptide, 400 mM ATP, 40 mM MgCl2, 1 mg/mL BSA, 50 mM Tris-HCl, pH 7.5, and 20–25 mCi/mL[g-32P] ATP—are added to initiate the reaction. There are two different substrate solutions in each. Following a 5-minute incubation period at 25°C, 20 mL of each mixture is spotted onto a phosphocellulose disc, which is then promptly submerged in 1% H3PO4. The peptide-incorporated 32P on the discs is counted in a scintillation counter after free [g-32P] ATP is removed. Following a five-minute incubation period at 25°C, 20 mL of each mixture is spotted onto a phosphocellulose disc, which is then promptly submerged in 1% H3PO4. Peptide-incorporated 32P on the discs is counted in a scintillation counter after free [g-32P] ATP is removed.
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| Cell Assay |
The MTT assay in microculture is used to estimate the cytostatic effect of daphnetin on MCF-7 tumor cells. The assay relies on the mitochondria of living cells reducing soluble tetrazolium salt. Dimethyl sulfoxide is used to dissolve the reduced product, an insoluble formazan with a purple hue, and measure it using spectrophotometry at 570 nm. The number of viable cells determines how much formazan forms. In each of the 96 microplate wells, 3 × 103 cells are seeded with a 200 μL medium containing the appropriate concentration of daphnetin. Five different concentrations of daphnetin are tested: 12.5 μM, 25 μM, 50 μM, 100 μM, and 200 μM. Following exposure for 24, 48, and 72 hours, the treated cells' percentage of proliferative inhibition is calculated in comparison to the control cells treated with solvent (PI% = [(T/C) − 1] × 100). Proliferation inhibition (PI); treated (T); and control (C). The least square concentration-response regressions are used to calculate the IC50.
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| Animal Protocol |
Immature CD1 (Im) female mice
35 mg/kg, 70 mg/kg, and 140 mg/kg Subcutaneously |
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| References |
| Molecular Formula |
C9H6O4
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| Molecular Weight |
178.14
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| Exact Mass |
178.03
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| Elemental Analysis |
C, 60.68; H, 3.40; O, 35.92
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| CAS # |
486-35-1
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| Appearance |
Solid powder
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| SMILES |
C1=CC(=C(C2=C1C=CC(=O)O2)O)O
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| InChi Key |
ATEFPOUAMCWAQS-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C9H6O4/c10-6-3-1-5-2-4-7(11)13-9(5)8(6)12/h1-4,10,12H
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| Chemical Name |
7,8-dihydroxychromen-2-one
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| Synonyms |
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| HS Tariff Code |
2934.99.03.00
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
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| 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) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (14.03 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 (14.03 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 (14.03 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 | 5.6136 mL | 28.0678 mL | 56.1356 mL | |
| 5 mM | 1.1227 mL | 5.6136 mL | 11.2271 mL | |
| 10 mM | 0.5614 mL | 2.8068 mL | 5.6136 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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