| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
Natural flavonoid
|
|---|---|
| ln Vitro |
2,3-Dehydrosilybin A and 2,3-dehydrosilybin B are a pair of enantiomers formed by the oxidation of the natural flavonolignans silybin A and silybin B, respectively. However, the antioxidant activity of 2,3-dehydrosilybin molecules is much stronger than that of their precursors. Here, we investigated the biotransformation of pure 2,3-dehydrosilybin A and 2,3-dehydrosilybin B in isolated human hepatocytes, and we also aimed to identify human UDP-glucuronosyltransferases (UGTs) and sulfotransferases (SULTs) with activity toward their respective enantiomers. After incubation with hepatocytes, both 2,3-dehydrosilybin A and 2,3-dehydrosilybin B were converted to hydroxyl derivatives, methylated hydroxyl derivatives, methyl derivatives, sulfates, and glucuronides. The products of direct conjugations predominated over those of oxidative metabolism, and glucuronides were the most abundant metabolites. Furthermore, we found that recombinant human UGTs 1A1, 1A3, 1A7, 1A8, 1A9, and 1A10 were capable of catalyzing the glucuronidation of both 2,3-dehydrosilybin A and 2,3-dehydrosilybin B. UGTs 1A1 and 1A7 showed the highest activity toward 2,3-dehydrosilybin A, and UGT1A9 showed the highest activity toward 2,3-dehydrosilybin B. The sulfation of 2,3-dehydrosilybin A and B was catalyzed by SULTs 1A1*1, 1A1*2, 1A2, 1A3, 1B1, 1C2, 1C4, and 1E1, of which SULT1A3 exhibited the highest activity toward both enantiomers. We conclude that 2,3-dehydrosilybin A and B are preferentially metabolized by conjugation reactions, and that several human UGT and SULT enzymes may play a role in these conjugations. [1]
|
| Enzyme Assay |
Incubation of 2,3-Dehydrosilybin A and B with Human Sulfotransferases [1]
Sulfation of the tested compounds by individual sulfotransferases was examined using cytosolic fractions from Escherichia coli expressing recombinant human SULTs 1A1*1, 1A1*2, 1A2, 1A3, 1B1, 1C2, 1C4, 1E1, or 2A1. The incubations were performed in 0.2 mL of potassium phosphate–HCl buffer (pH 7.4; 50 mM) containing 5 mM MgCl2, 10 mM dithiothreitol, 50 µM 2,3-dehydrosilybin A or B (in 0.1% (v/v) DMSO), 50 µg/mL E. coli cytosol protein, and 120 µM 3′-phosphoadenosine 5′-phosphosulfate (PAPS). Control samples were prepared by incubating the tested compounds in the absence of PAPS and/or the cytosolic fraction. All samples were incubated for 30 min at 37 °C and 300 rpm in a Thermomixer Comfort, and then stored at –80 °C until their analysis by UHPLC-MS. |
| Cell Assay |
Incubation of 2,3-Dehydrosilybin A and B with Human Hepatocytes [1]
The samples of human liver were obtained from multiorgan donors. Hepatocytes were isolated using two-step collagenase perfusion, and resuspended in serum-free medium containing Williams’ medium E, Ham’s F-12 medium, and additives as described previously. Hepatocyte cultures used in the study were prepared from liver samples of three donors: a 45-year-old man (culture LH81), a 56-year-old man (culture LH83), and a 47-year-old woman (culture LH84). Suspensions of human hepatocytes (4 × 106 cell/mL) were incubated with 0.1% (v/v) DMSO (control) or with 50 µM 2,3-dehydrosilybin A or B (in 0.1% (v/v) DMSO). After 1 h of incubation at 37 °C and 160 rpm in an ES-20 Environmental Shaker, the cells and media were separated by centrifugation for 5 min at 150× g and 4 °C, and stored at −80 °C until their analysis by ultra-high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS). Incubation of 2,3-Dehydrosilybin A and B with Human UGTs [1] Glucuronidation of the tested compounds by individual UGT enzymes was examined using Corning Supersomes, i.e., microsomes from baculovirus-transfected insect cells expressing the recombinant human UGTs 1A1, 1A3, 1A4, 1A6, 1A7, 1A8, 1A9, 1A10, 2B4, 2B7, 2B10, 2B15, or 2B17. The incubations were performed in 0.25 mL of Tris–HCl buffer (pH 7.4; 100 mM) containing 8 mM MgCl2, 25 µg/mL alamethicin, 50 µM 2,3-dehydrosilybin A or B, 0.2 mg/mL microsomal protein and 2 mM UDP-glucuronic acid, with a final concentration of DMSO of 0.6% (v/v). Control samples were prepared by incubating the tested compounds with Corning Supersomes insect cell control microsomes, which lack UGT activity. All samples were incubated for 30 min at 37 °C and 300 rpm in a Thermomixer Comfort, and then stored at −80 °C until their analysis by UHPLC-MS. |
| References |
| Molecular Formula |
C25H20O10
|
|---|---|
| Exact Mass |
480.105
|
| CAS # |
142796-24-5
|
| Related CAS # |
2,3-Dehydrosilybin A;25166-14-7
|
| PubChem CID |
12051803
|
| Appearance |
Light yellow to yellow solid powder
|
| Density |
1.574±0.06 g/cm3(Predicted)
|
| Boiling Point |
761.0±60.0 °C(Predicted)
|
| LogP |
3.1
|
| Hydrogen Bond Donor Count |
5
|
| Hydrogen Bond Acceptor Count |
10
|
| Rotatable Bond Count |
4
|
| Heavy Atom Count |
35
|
| Complexity |
816
|
| Defined Atom Stereocenter Count |
2
|
| SMILES |
O=C1C(O)=C(OC=2C=C(O)C=C(O)C12)C=3C=CC=4OC(CO)C(OC4C3)C5=CC=C(O)C(OC)=C5
|
| InChi Key |
BVKQRAYKLBRNIK-RDPSFJRHSA-N
|
| InChi Code |
InChI=1S/C25H20O10/c1-32-17-6-11(2-4-14(17)28)24-20(10-26)33-16-5-3-12(7-18(16)34-24)25-23(31)22(30)21-15(29)8-13(27)9-19(21)35-25/h2-9,20,24,26-29,31H,10H2,1H3/t20-,24-/m0/s1
|
| Chemical Name |
3,5,7-trihydroxy-2-[(2S,3S)-3-(4-hydroxy-3-methoxyphenyl)-2-(hydroxymethyl)-2,3-dihydro-1,4-benzodioxin-6-yl]chromen-4-one
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| Solubility (In Vitro) |
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
|
|---|---|
| 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.) |
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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