| Size | Price | Stock | Qty |
|---|---|---|---|
| 500mg |
|
| Targets |
- Cell adhesion-related molecules (vinculin, F-actin) (No IC50/Ki/EC50 data available; modulates the expression and polymerization of these molecules to affect cell-substrate attachment) [1]
|
|---|---|
| ln Vitro |
5a-Pregnane-3,20-dione reduces the expression of vinculin, adhesion plaques, and cell-substrate attachment. In MCF-7 breast cancer cells, F-actin[1].
- In MCF-7 breast cancer cells, 5a-Pregnane-3,20-dione (10-1000 nM) decreased cell-substrate attachment in a concentration- and time-dependent manner. At 100 nM (24 h treatment), cell adhesion rate reduced by 42% compared to the control group (measured by crystal violet staining and absorbance detection at 570 nm); at 1000 nM (48 h treatment), the adhesion rate further decreased by 68% [1] - The drug reduced the number of adhesion plaques in MCF-7 cells. Immunofluorescence staining showed that 100 nM 5a-Pregnane-3,20-dione (24 h) decreased the average number of adhesion plaques per cell from 18 to 7, with a reduction rate of 61% [1] - It downregulated vinculin expression in MCF-7 cells. Western blot analysis revealed that 100-1000 nM 5a-Pregnane-3,20-dione (48 h) reduced vinculin protein levels in a concentration-dependent manner: 100 nM caused a 35% reduction, while 1000 nM caused a 72% reduction (normalized to β-actin) [1] - 5a-Pregnane-3,20-dione inhibited F-actin polymerization. Phalloidin staining showed that 100 nM (24 h) treatment decreased the fluorescence intensity of polymerized F-actin by 53% compared to the control, indicating disrupted actin cytoskeleton structure [1] |
| Cell Assay |
- Cell-substrate attachment assay: MCF-7 cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum. 96-well plates were pre-coated with type I collagen (10 μg/mL) and air-dried. Cells (5×10⁴ cells/well) were seeded and treated with 5a-Pregnane-3,20-dione (10-1000 nM) for 24-48 h. Non-adherent cells were removed by washing with PBS, and adherent cells were fixed with 4% paraformaldehyde, stained with 0.1% crystal violet, and dissolved in 10% acetic acid. Absorbance was measured at 570 nm to calculate adhesion rate [1]
- Adhesion plaque observation assay: MCF-7 cells were plated on glass coverslips pre-coated with type I collagen (10 μg/mL) and treated with 5a-Pregnane-3,20-dione (100 nM) for 24 h. Cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100, and incubated with anti-vinculin primary antibody and fluorescent secondary antibody. Adhesion plaques were observed under a confocal laser scanning microscope, and the number of plaques per cell was counted (n=50 cells per group) [1] - Vinculin Western blot assay: MCF-7 cells were treated with 5a-Pregnane-3,20-dione (100-1000 nM) for 48 h. Cells were lysed with RIPA buffer containing protease inhibitors, and protein concentration was determined by BCA assay. Equal amounts of protein (30 μg/lane) were separated by SDS-PAGE, transferred to PVDF membranes, and incubated with anti-vinculin and anti-β-actin primary antibodies, followed by HRP-conjugated secondary antibodies. Bands were visualized by ECL reagent, and band intensity was quantified using image analysis software [1] - F-actin staining assay: MCF-7 cells on glass coverslips were treated with 5a-Pregnane-3,20-dione (100 nM) for 24 h, fixed with 4% paraformaldehyde, and stained with rhodamine-conjugated phalloidin (1:100 dilution) for 30 min. Nuclei were counterstained with DAPI. Fluorescence intensity of F-actin was measured using image analysis software (n=30 cells per group) [1] |
| References | |
| Additional Infomation |
5α-Pregnane-3,20-dione is a C21 steroid hormone formed by the carbonyl substitution of 5α-pregnane at the 3 and 20 positions. It is a metabolite of progesterone and functions as both a metabolite and a progestin in the human body. It is a 20-oxosteroid, a C21 steroid, and a 3-oxo-5α-steroid, functionally related to progesterone, and derived from the hydride of 5α-pregnane. 5α-Pregnane-3,20-dione has been reported and data are available in both Homo sapiens and Pilocarpus. It is a biologically active 5α-reduced metabolite of plasma progesterone. It is a direct precursor of 5α-pregnane-3α-ol-20-one (allogrenolone), a neuroactive steroid that binds to GABA(A) receptors.
- 5α-pregnane-3,20-dione is an endogenous metabolite of progesterone, mainly produced by the adrenal glands and gonads[1] - The mechanism by which it reduces MCF-7 cell adhesion to the matrix involves downregulating the expression of vinculin, a key component of focal adhesion, and inhibiting F-actin polymerization, both of which disrupt the adhesion structure and the formation of the actin cytoskeleton[1] - This study suggests that 5α-pregnane-3,20-dione may affect the migration and invasion of MCF-7 breast cancer cells by impairing cell adhesion, although no direct migration/invasion experiments were performed in this study. [1] |
| Molecular Formula |
C21H32O2
|
|---|---|
| Molecular Weight |
316.4776
|
| Exact Mass |
316.24
|
| CAS # |
566-65-4
|
| PubChem CID |
92810
|
| Appearance |
White to off-white solid powder
|
| Density |
1.0±0.1 g/cm3
|
| Boiling Point |
429.2±38.0 °C at 760 mmHg
|
| Melting Point |
204 °C
|
| Flash Point |
160.3±23.8 °C
|
| Vapour Pressure |
0.0±1.0 mmHg at 25°C
|
| Index of Refraction |
1.516
|
| LogP |
4.31
|
| Hydrogen Bond Donor Count |
0
|
| Hydrogen Bond Acceptor Count |
2
|
| Rotatable Bond Count |
1
|
| Heavy Atom Count |
23
|
| Complexity |
538
|
| Defined Atom Stereocenter Count |
7
|
| SMILES |
CC(=O)[C@H]1CC[C@@H]2[C@@]1(CC[C@H]3[C@H]2CC[C@@H]4[C@@]3(CCC(=O)C4)C)C
|
| InChi Key |
XMRPGKVKISIQBV-BJMCWZGWSA-N
|
| InChi Code |
InChI=1S/C21H32O2/c1-13(22)17-6-7-18-16-5-4-14-12-15(23)8-10-20(14,2)19(16)9-11-21(17,18)3/h14,16-19H,4-12H2,1-3H3/t14-,16-,17+,18-,19-,20-,21+/m0/s1
|
| Chemical Name |
(5S,8R,9S,10S,13S,14S,17S)-17-acetyl-10,13-dimethyl-1,2,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydrocyclopenta[a]phenanthren-3-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) |
DMSO : ~6.67 mg/mL (~21.08 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 0.67 mg/mL (2.12 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 6.7 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.1598 mL | 15.7988 mL | 31.5976 mL | |
| 5 mM | 0.6320 mL | 3.1598 mL | 6.3195 mL | |
| 10 mM | 0.3160 mL | 1.5799 mL | 3.1598 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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