| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
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| Other Sizes |
| Targets |
Epoxygenase (cytochrome P450 enzymes CYP1A1, CYP2B1, CYP2J3) – indirect activation leading to accumulation of epoxyeicosanoids. [1]
Intracellular calcium stores (ryanodine-sensitive) – induces Ca2+ release from intracellular stores. [2] |
|---|---|
| ln Vitro |
In C6 rat glioma cells, Saikogenin D (1–20 μM) has an IC50 value of 3 μM and has a concentration-dependent effect on PGE2 synthesis caused by the Ca2+ ionophore A23187 [1]. In C6 rat glioma cells, as long as extracellular Ca2+ is present or absent, saikogenin D (10-100 μM) raises [Ca2+]i in a concentration-dependent manner with an EC50 value of 35 μM [1].
Saikogenin D (1–20 µM) inhibited A23187-induced PGE2 production in C6 rat glioma cells in a concentration-dependent manner with an IC50 of approximately 3 µM. It also inhibited histamine (100 µM)-induced PGE2 production. [2] Saikogenin D (20 µM for 10 min) did not affect basal PGE2 release but inhibited A23187-induced PGE2 production. [1] Saikogenin D (50 µM) facilitated A23187-induced production of 11,12-dihydroxyeicosatrienoic acid (DHET), a metabolite of 11,12-epoxyeicosatrienoic acid (EET), in C6 cells labeled with [14C]-arachidonic acid. [1] Saikogenin D (20 or 50 µM for 20 min) did not affect mRNA expression of CYP1A1, CYP2B1, or CYP2J3 in C6 cells as measured by real-time PCR. [1] Saikogenin D did not affect the conversion of arachidonic acid to PGE2 in microsomal preparations (up to 30 µM tested, no direct COX inhibition). [2] Saikogenin D did not affect arachidonic acid liberation from intact C6 cells (concentrations up to 10 µM; A23187-induced release taken as 100%, saikogenin D at 2, 5, 10 µM gave 108.9%, 116.7%, 117.0% respectively, not statistically significant vs control). [2] Saikogenin D (10 µM) did not affect the [Ca2+]i elevation induced by 1 µM ionomycin. [2] Saikogenin D (10–100 µM) elevated intracellular free Ca2+ concentration ([Ca2+]i) in a concentration-dependent manner with an EC50 of approximately 35 µM in the presence or absence of extracellular Ca2+ (in the presence of 4 mM EGTA), indicating Ca2+ release from intracellular stores. [2] |
| Enzyme Assay |
Cyclooxygenase (COX) activity assay: Recombinant COX-1 or COX-2 enzyme protein (2.0 U each) was dissolved in Tris-buffer containing haematin and tryptophan. The mixture was pre-incubated with or without Saikogenin D for 10 minutes at 37°C, then further incubated with 20 µM arachidonic acid for 10 minutes at 37°C. The reaction was terminated by adding HCl. PGE2 was extracted and measured by radioimmunoassay. Saikogenin D (up to 30 µM) did not inhibit COX-1 or COX-2 activity (indomethacin showed inhibition). [1]
Microsomal COX activity assay: Microsomes (90 µg/tube) from C6 cells were incubated with various concentrations of Saikogenin D for 10 minutes, then further incubated with 20 µM arachidonic acid for 40 minutes. Biosynthesized PGE2 was determined by radioimmunoassay. Saikogenin D did not inhibit the conversion of arachidonic acid to PGE2. [2] |
| Cell Assay |
PGE2 production measurement: C6 rat glioma cells were cultivated in 12-well plates. Two days after seeding, cells were washed with HEPES-buffered Eagle's minimum essential medium. After drug treatment (Saikogenin D pre-incubation for 10 min, then with or without A23187 for 10 min), the medium was retrieved and indomethacin and EDTA were added to terminate COX reaction. PGE2 was extracted twice with ethyl acetate under acidic conditions (pH 4.0), dried under nitrogen, dissolved in Tris-HCl buffer, and measured by radioimmunoassay. [1][2]
Arachidonic acid metabolite analysis: C6 cells were seeded into 6-well plates and labeled with [14C]-arachidonic acid for 24 h. Cells were washed, pre-incubated with Saikogenin D for 10 min, then incubated with or without A23187 for 10 min. The medium was collected, COX reaction terminated, metabolites extracted with ethyl acetate, dried, dissolved in chloroform, and separated by thin-layer chromatography using ethyl acetate-isooctane-acetic acid-water upper phase. Radioluminogram was visualized with a molecular imager. [1] RT-PCR for epoxygenase mRNA: Total RNA was prepared from C6 cells using ISOGEN, reverse-transcribed into cDNA using oligo(dT) primer and reverse transcriptase. PCR was performed using specific primers for CYP1A1, CYP2B1, CYP2B2, and CYP2J3. PCR products were separated on 1.5% agarose gel and stained with ethidium bromide. Real-time PCR was also performed using SYBR Premix Ex Taq with β-actin as endogenous control. Saikogenin D (20 or 50 µM for 20 min) did not alter mRNA expression levels. [1] Intracellular calcium measurement: C6 cells were loaded with fura-2 acetoxymethyl ester. [Ca2+]i was monitored by fluorescence spectrophotometry. Cells were incubated with Saikogenin D (5 min pre-incubation, then stimulated with ionomycin, or direct concentration-response) in the presence or absence of extracellular Ca2+ (with 4 mM EGTA). [2] Arachidonic acid release assay: C6 cells labeled with [14C]-arachidonic acid were incubated with Saikogenin D for 10 min, then with or without A23187 for an additional 10 min. Released radioactivity was measured by liquid scintillation counting. [2] |
| References |
|
| Additional Infomation |
According to reports, plants in the genus Bupleurum contain saikosaponin D, and there is relevant data on the presence of this substance in plants in the genus Bupleurum.
Saikogenin D is an intestinal metabolite of saikosaponin D; when saikosaponin D is orally administered, it is transformed to saikogenin D before absorption into the blood. [2] Saikogenin D inhibits PGE2 production not by direct COX inhibition but by activating epoxygenase pathway, leading to accumulation of 11,12-EET and 11,12-DHET, which secondarily inhibit COX-2 activity. [1] The IC50 for inhibition of A23187-induced PGE2 production is about 3 µM, while the EC50 for [Ca2+]i elevation is about 35 µM, suggesting that at low concentrations the anti-inflammatory effect predominates. [2] |
| Molecular Formula |
C30H48O4
|
|---|---|
| Molecular Weight |
472.6997
|
| Exact Mass |
472.355
|
| CAS # |
5573-16-0
|
| PubChem CID |
9890994
|
| Appearance |
White to off-white solid
|
| Density |
1.2±0.1 g/cm3
|
| Boiling Point |
607.4±55.0 °C at 760 mmHg
|
| Melting Point |
261-266 °C
|
| Flash Point |
252.4±26.1 °C
|
| Vapour Pressure |
0.0±3.9 mmHg at 25°C
|
| Index of Refraction |
1.586
|
| LogP |
5.38
|
| Hydrogen Bond Donor Count |
4
|
| Hydrogen Bond Acceptor Count |
4
|
| Rotatable Bond Count |
2
|
| Heavy Atom Count |
34
|
| Complexity |
921
|
| Defined Atom Stereocenter Count |
9
|
| SMILES |
O([H])[C@@]1([H])C([H])([H])C([H])([H])[C@@]2(C([H])([H])[H])[C@@]([H])(C([H])([H])C([H])([H])[C@@]3(C([H])([H])[H])[C@]4(C([H])([H])[H])C([H])([H])[C@]([H])([C@@]5(C([H])([H])O[H])C([H])([H])C([H])([H])C(C([H])([H])[H])(C([H])([H])[H])C([H])([H])C5=C4C([H])=C([H])[C@@]32[H])O[H])[C@]1(C([H])([H])[H])C([H])([H])O[H]
|
| InChi Key |
QGNVMEXLLPGQEV-IULQVHCXSA-N
|
| InChi Code |
InChI=1S/C30H48O4/c1-25(2)13-14-30(18-32)20(15-25)19-7-8-22-26(3)11-10-23(33)27(4,17-31)21(26)9-12-28(22,5)29(19,6)16-24(30)34/h7-8,21-24,31-34H,9-18H2,1-6H3/t21-,22-,23+,24-,26+,27+,28-,29-,30-/m1/s1
|
| Chemical Name |
(3S,4R,4aR,6aR,6bS,8R,8aS,14aR,14bS)-4,8a-bis(hydroxymethyl)-4,6a,6b,11,11,14b-hexamethyl-1,2,3,4a,5,6,7,8,9,10,12,14a-dodecahydropicene-3,8-diol
|
| 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 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)
|
| 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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1155 mL | 10.5775 mL | 21.1551 mL | |
| 5 mM | 0.4231 mL | 2.1155 mL | 4.2310 mL | |
| 10 mM | 0.2116 mL | 1.0578 mL | 2.1155 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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