S6K1
Eudesmin targets S6K1 (ribosomal protein S6 kinase 1) [1].
Eudesmin inhibits the activity and nuclear translocation of S6K1 [1].

Eudesmin (20, 40, and 80μM) treatment of mesenchymal stem cells (MSCs) inhibits the S6K1 signaling pathway, which prevents adipogenesis. Nuclear translocation and S6K1 activation are inhibited by eudesmin treatment. Following Eudesmin treatment, there is a decrease in the phosphorylation of H2B serine 36 (H2BS36p) mediated by S6K1 [1].

---

In C3H10T1/2 mouse mesenchymal stem cells (MSCs), treatment with eudesmin (20-80 µM for 24 hours) abolished phosphorylation of downstream S6K1 substrates, ribosomal protein S6 (S235/236) and H2B (S36), in a dose-dependent manner [1].
Treatment with eudesmin (80 µM for 24 hours) decreased the level of nuclear S6K1 and increased cytoplasmic S6K1, and reduced phosphorylation of S6 in both subcellular locations and H2B in the nucleus [1].
Eudesmin (80 µM for 24 hours) abrogated the enrichment of H2BS36 phosphorylation (H2BS36p) at the promoter regions of Wnt6, Wnt10a, and Wnt10b genes as shown by ChIP-qPCR [1].
Eudesmin (80 µM for 24 hours) significantly increased the mRNA expression of Wnt6, Wnt10a, and Wnt10b genes as measured by qPCR [1].
During adipogenic commitment (4 days with BMP4), eudesmin (80 µM) increased the mRNA levels of Wnt6, Wnt10a, and Wnt10b and inhibited the expression of adipogenic transcription factors PPARγ and Cebpa in 10T1/2 cells [1].
Eudesmin (80 µM) treatment during the adipogenic commitment stage, terminal differentiation stage, or the whole differentiation stage prevented lipid accumulation in terminally differentiated adipocytes, as visualized by Oil Red O staining [1].
Treatment of 10T1/2 cells with eudesmin (80 µM for 24 hours) increased the protein levels of MyoD (myogenesis marker) and Runx2 (osteogenesis marker) [1].
Eudesmin (80 µM for 24 hours) significantly increased mRNA levels of myogenic genes (Myf5, MyoD, Pax3) and osteogenic genes (Bmp2, Col1a1, Sp7, Runx2) in 10T1/2 cells [1].

C3H10T1/2 mouse mesenchymal stem cells were grown in DMEM supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin [1]. For adipogenic differentiation, cells were incubated in DMEM with 10 µg/ml BMP4, 10% FBS, and 1% P/S for 4 days for commitment to preadipocytes [1]. For terminal differentiation, pre-adipocytes were further incubated in DMEM with 10% FBS, 1% P/S, 0.5 mM IBMX, 1 µM dexamethasone, and 1 µg/ml insulin, followed by medium changes with DMEM containing 10% FBS, 1% P/S, and 1 µg/ml insulin every other day [1].
For immunoblotting, cells were lysed in Pro-Prep buffer, proteins were subjected to SDS-PAGE, transferred to PVDF membranes, incubated overnight with primary antibodies, then with HRP-conjugated secondary antibodies for 1 hour, and signals were detected using chemiluminescence reagents and quantified with ImageJ [1].
For subcellular fractionation, cells were resuspended in Buffer A (10 mM HEPES, 1.5 mM MgCl2, 10 mM KCl, 1 mM EDTA, 1 mM DTT, 0.5 µg/ml leupeptin, 1 mM PMSF, 1 µM pepstatin A, 0.05% NP-40), cytoplasmic extracts were separated by centrifugation at 3000 rpm and 4°C for 10 min [1]. The pellet was resuspended in Buffer B (20 mM HEPES, 1.5 mM MgCl2, 420 mM KCl, 25% glycerol, 0.2 mM EDTA, 1 mM DTT, 0.5 µg/ml leupeptin, 1 mM PMSF, 1 µM pepstatin A), incubated on ice for 30 min, and nuclear extracts were separated by centrifugation at 13000 rpm and 4°C for 20 min [1].
For qPCR, total RNA was extracted using Easy-Blue reagent, 1 µg of total RNA was reverse transcribed into cDNA using a Maxim RT-PreMix Kit, and qPCR was performed using KAPA SYBR FAST qPCR Master Mix and a CFX96 Touch real-time PCR detector [1]. Relative mRNA levels were normalized to β-actin [1].
For chromatin immunoprecipitation (ChIP), cross-linked and sheared chromatin was immunoprecipitated overnight at 4°C with appropriate antibodies, and recovered chromatin fragments were subjected to qPCR using primer pairs specific for the target gene promoter [1].
For Oil Red O staining, fully differentiated adipocytes were fixed with 10% formalin for 1 hour, washed with 60% isopropanol, incubated with Oil Red O working solution for 1 hour, and rinsed with distilled water three times [1]. The Oil Red O stock solution was prepared by dissolving 300 mg of Oil Red O powder in 100 ml of 99% isopropanol, and working solution was prepared by diluting 30 ml of stock solution with 20 ml of distilled water just before use [1].

[1]. Eudesmin Impairs Adipogenic Differentiation via Inhibition of S6K1 Signaling Pathway. Biochem Biophys Res Commun. 2018 Nov 10;505(4):1148-1153.

(3R,3aS,6R,6aS)-3,6-bis(3,4-dimethoxyphenyl)-1,3,3a,4,6,6a-hexahydrofurano[3,4-c]furan has been reported in Stellera chamaejasme, Metrodorea nigra, and other organisms with available data.

---

Eudesmin is a novel small molecule that blocks adipogenesis through down-regulation of the S6K1-H2BS36p axis, followed by regulation of cell fate determination genes [1]. The study suggests a promising therapeutic approach with eudesmin to treat obesity and metabolic diseases [1]. S6K1-knockout mice exhibit resistance to high fat diet-induced obesity due to increased insulin sensitivity, enhanced β-oxidation, and impaired adipogenesis [1]. Upon adipogenic stimulus, S6K1 is activated and translocated to the nucleus to phosphorylate H2B at serine 36, suppressing the expression of Wnt6, Wnt10a, and Wnt10b, which disrupt adipogenesis [1]. Eudesmin inhibits S6K1-mediated H2BS36 phosphorylation, leading to increased expression of Wnt genes and promotion of myogenic and osteogenic gene expression in MSCs [1].

C22H26O6

386.4382

386.172

526-06-7

325601

White to off-white solid

1.2±0.1 g/cm3

517.3±50.0 °C at 760 mmHg

108.5-109.5℃

209.8±30.0 °C

0.0±1.3 mmHg at 25°C

1.547

3.08

0

6

6

28

457

4

O1C([H])([H])C2([H])C([H])(C3C([H])=C([H])C(=C(C=3[H])OC([H])([H])[H])OC([H])([H])[H])OC([H])([H])C2([H])C1([H])C1C([H])=C([H])C(=C(C=1[H])OC([H])([H])[H])OC([H])([H])[H]

PEUUVVGQIVMSAW-DJDZNOHASA-N

InChI=1S/C22H26O6/c1-23-17-7-5-13(9-19(17)25-3)21-15-11-28-22(16(15)12-27-21)14-6-8-18(24-2)20(10-14)26-4/h5-10,15-16,21-22H,11-12H2,1-4H3/t15-,16-,21+,22+/m1/s1

(3R,3aS,6R,6aS)-3,6-bis(3,4-dimethoxyphenyl)-1,3,3a,4,6,6a-hexahydrofuro[3,4-c]furan

Eudesmin

2934.99.9001

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.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~100 mg/mL (~258.77 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.47 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 (6.47 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 (6.47 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 25.0 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.)