| Size | Price | Stock | Qty |
|---|---|---|---|
| 10mg |
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| 1g | |||
| Other Sizes |
| Targets |
Gymnestrogenin (compound 11) is a dual antagonist for Liver X Receptor \(\alpha\) (LXRα) and Liver X Receptor \(\beta\) (LXRβ). [2]
IC50 for LXRα: 2.5 μM. [2] IC50 for LXRβ: 1.4 μM. [2] |
|---|---|
| ln Vitro |
In vitro, estrogen suppresses SREBP1c and ABCA1 expression and can lessen lipid build-up in HepG2 cells [2].
Gymnestrogenin (11) at 10 μM reduced the basal transactivation of both LXRα and LXRβ in a reporter gene assay using HepG2 cells (p < 0.05 versus not treated cells). [2] When co-administered with 10 μM GW3965 (a dual LXRα/β agonist), Gymnestrogenin (11) at 50 μM antagonized the effect of GW3965 on LXRα transactivation (p < 0.05). [2] Gymnestrogenin (11) at 50 μM failed to antagonize the transcriptional activity driven by FXR, GP-BAR1, PXR, and PPARγ. [2] Gymnestrogenin (11) at 10 μM reduced the relative mRNA expression of SREBP-1c in HepG2 cells both under basal conditions and in cells co-stimulated with 10 μM GW3965 (p < 0.05 versus not treated cells; p < 0.05 versus GW3965 stimulated cells). [2] Gymnestrogenin (11) at 10 μM reduced the expression of ABCA1 in THP1 cells co-stimulated with 10 μM GW3965 (p < 0.05 versus GW3965 stimulated cells). [2] Gymnestrogenin (11) at 10 μM abrogated the effects of 10 μM GW3965 on intracellular lipid accumulation in HepG2 cells measured by Oil Red O staining, significantly reducing red lipid droplets inside cells. Quantitative analysis confirmed the data to be statistically significant. [2] Gymnestrogenin (11) at 10 μM reduced the relative mRNA expression of LXRβ (p < 0.05 versus GW3965 stimulated cells) but not that of LXRα in HepG2 cells stimulated with 10 μM GW3965. [2] |
| Enzyme Assay |
The study used molecular modeling to predict interactions. The induced fit docking protocol (IFD) was used to calculate movements of backbone and side chains upon ligand binding to the ligand binding domain (LBD) of LXR isoforms. For LXRβ, the docked pose of Gymnestrogenin (11) showed van der Waals contacts with residues Ser242, Phe268, Phe271, Thr272, Leu274, Ala275, Ile277, Ser278, Ile309, Met312, Glu315, Ile327, Thr328, Phe329, Leu330, Phe340, Leu345, Phe349, His435, Gln438, Val439, Leu442, Leu449, Leu453, and Trp457. The double bond at C-12 is parallel to the side chain of Phe315. The hydroxyl group at C-3 donates a hydrogen bond to the side chain of Gln438. The OH at C-23 donates an H-bond to Gln438 and at C-28 interacts with the backbone CO of Phe257. The OH at C-16 accepts two H-bonds from the side chain of Arg232. The OH at C-21 interacts with the side chain of Arg319. [2]
|
| Cell Assay |
For the LXR transactivation assay, HepG2 cells were plated at 5×10^4 cells/well in a 24-well plate. Cells were transfected with 200 ng of reporter vector p(UAS)5xTKLuc, 100 ng of a vector containing the ligand binding domain of LXRα or LXRβ cloned upstream of the GAL4-DNA binding domain (pSG5-LXRαLBD-GAL4DBD or pSG5-LXRβLBD-GAL4DBD), and 100 ng of pGL4.70 (a vector encoding the human Renilla gene). At 24 h post-transfection, cells were stimulated for 18 h with 10 μM GW3965 (a dual LXRα/β agonist), with test compounds (1-11) at 10 μM or 50 μM, or with combinations. After treatments, 10 μL of cellular lysates were read using a Dual Luciferase Reporter Assay System. [2]
For the specificity assay against GPBAR1, HEK-293T cells were plated at 1×10^4 cells/well in a 24 well-plate and transfected with 200 ng of pGL4.29 (a reporter vector containing a cAMP response element, CRE), with 100 ng of pCMVSPORT6-human GP-BAR1, and with 100 ng of pGL4.70. [2] For Real-Time PCR, total RNA was isolated using TRlzol reagent. One μg RNA was purified of genomic DNA by DNase I treatment and random reverse-transcribed. Ten ng of template cDNA was added to a PCR mixture containing 0.2 μM of each primer and SYBR FAST Universal ready mix. All reactions were performed in triplicate. The relative mRNA expression was calculated and expressed as 2^-(ΔΔCT). [2] For Oil Red O (ORO) staining, HepG2 cells were grown at an initial density of 10^5 cells/well in a 6-well plate and treated daily for 7 d with 10 μM GW3965 or with the combination of GW3965 plus Gymnestrogenin (11) at 10 μM. Cells were then washed, fixed with 10% formalin for 60 min, washed with dH2O and 60% isopropanol, and then stained with Oil Red O working solution for 15 min at room temperature. Cells were washed and acquired under microscope for qualitative analysis. To quantify Oil Red O content, it was eluted by incubating cells with 100% isopropanol for 10 min, and the density of samples was read at 500 nm. [2] |
| References | |
| Additional Infomation |
Reports indicate that Gymnema sylvestre contains gymnema estrogen, and relevant data is available for reference.
Gymnestrogenin (M = 490) is a new pentahydroxy-triterpene from Gymnema sylvestre leaves, distinct from Gymnemagenin. Unlike Gymnemagenin, Gymnestrogenin shows a negative reaction with benzidine-periodate, suggesting it does not contain a 1,2-diol grouping. Acetylation of Gymnestrogenin with acetic anhydride and pyridine at 35°C for 16 hours yielded an amorphous but uniform Penta-O-acetyl-gymnestrogenin derivative (C40H60O10), indicating all hydroxy groups are easily acetylable. [1] The proposed structure of Gymnestrogenin (3) is based on spectroscopic data, the easy acetylatability of the hydroxy groups, and its relationship to Gymnemagenin, and thus remains hypothetical and requires verification through chemical experiments. [1] Gymnestrogenin (11) is a less common aglycone in gymnemic acids, differing from the widespread aglycone gymmenagenin (10) by the absence of a hydroxyl group on ring E (position 22). This structural difference results in a dual LXRα/β antagonistic profile for Gymnestrogenin, whereas gymmenagenin is a selective LXRβ antagonist. [2] The study suggests that the presence of the hydroxyl group at position 22 is a negative factor in LXRβ recognition by non-acylated gymnemic acids, and its absence in Gymnestrogenin allows for dual antagonism. [2] Gymnestrogenin (11) could reduce the transcriptional activity of LXR on its own promoter, thus reducing the mRNA expression of LXRβ. [2] |
| Molecular Formula |
C30H50O5
|
|---|---|
| Exact Mass |
490.366
|
| CAS # |
19942-02-0
|
| PubChem CID |
15560302
|
| Appearance |
White to off-white solid powder
|
| Melting Point |
288-289 °C
|
| LogP |
4.055
|
| Hydrogen Bond Donor Count |
5
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
2
|
| Heavy Atom Count |
35
|
| Complexity |
899
|
| Defined Atom Stereocenter Count |
11
|
| SMILES |
C[C@]12CC[C@@H]([C@@]([C@@H]1CC[C@@]3([C@@H]2CC=C4[C@]3(C[C@@H]([C@@]5([C@H]4CC([C@H](C5)O)(C)C)CO)O)C)C)(C)CO)O
|
| InChi Key |
SIBYGGBNBRCVQI-DGNDGBPUSA-N
|
| InChi Code |
InChI=1S/C30H50O5/c1-25(2)13-19-18-7-8-21-26(3)11-10-22(33)27(4,16-31)20(26)9-12-28(21,5)29(18,6)14-24(35)30(19,17-32)15-23(25)34/h7,19-24,31-35H,8-17H2,1-6H3/t19-,20+,21+,22-,23-,24-,26-,27-,28+,29+,30+/m0/s1
|
| Chemical Name |
(3S,4aS,5S,6aR,6aS,6bR,8aR,9R,10S,12aR,14bS)-4a,9-bis(hydroxymethyl)-2,2,6a,6b,9,12a-hexamethyl-1,3,4,5,6,6a,7,8,8a,10,11,12,13,14b-tetradecahydropicene-3,5,10-triol
|
| 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)
|
| 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
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|---|---|
| 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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