Natural product/Steroids; VEGF-VEGF-R2-Akt
(E)-Guggulsterone targets multiple receptors and enzymes. It is an antagonist of the farnesoid X receptor (FXR) with IC50 values of 15 µM (for CDCA-induced FXR activation) and 24.06 µM. It binds to mineralocorticoid receptor (MR) with Ki ≈ 35 nM, which is >100 times more potent than its affinity for FXR. It also binds to androgen receptor (AR), glucocorticoid receptor (GR), and progesterone receptor (PR) with Ki values ranging from 224 to 315 nM. In cell-based assays, (E)-Guggulsterone acts as an antagonist of AR, GR, and MR, but as an agonist of PR, with very low agonist activity on estrogen receptor alpha (ERα, EC50 > 5000 nM). It also inhibits CYP2C9 (19 μM) and CYP2C19 (2.1 μM).

GugguLsterone (0.5 -20 μM; 24 hours) suppresses the production of TREM-1, TLR4 and TNF-α and the phosphorylation of IκBα and NF-κB p65 by LPS [2].

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Guggulsterone suppressed LX-2 cell growth in a dose- and activation-dependent manner. This growth suppression was due to the induction of HSC apoptosis, which was mediated by the activation of c-Jun N-terminal kinase and mitochondrial apoptotic signaling. Additionally, guggulsterone regulated phosphorylation of Akt and adenosine monophosphate-activated protein kinase, which were subsequently proven responsible for the guggulsterone-induced HSC growth suppression. Guggulsterone inhibited NF-κB activation in LX-2 cells, which is one of the major mediators in HSC activation. Indeed, guggulsterone decreased collagen α1 synthesis and α-smooth muscle actin expression in these cells. [3]

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(E)-Guggulsterone inhibits the growth of a wide variety of tumor cells and induces apoptosis through downregulation of antiapoptotic gene products (IAP1, xIAP, Bfl-1/A1, Bcl-2, cFLIP and survivin), modulation of cell cycle proteins (cyclin D1 and c-Myc), activation of caspases and JNK, and inhibition of Akt. It decreases CDCA-induced FXR activation with IC50s of 15 μM for E-guggulsterone. In human breast epithelial MCF10A cells, (E)-Guggulsterone (5-25 μM, 0-12 hours) activates the Nrf2 signaling pathway, induces HO-1 expression, and leads to moderate intracellular ROS accumulation. In Huh-7 cells, it exhibits potent anti-dengue virus activity (0-20 μM, 3 days) through Nrf2/HO-1 pathway activation and restoration of antiviral interferon responses. It shows antibacterial effects against Bacillus subtilis, Staphylococcus aureus, and Pseudomonas aeruginosa (3.2 mM, 24 hours) with inhibition zone diameters of 14 mm, 14 mm, and 11 mm, respectively. In mouse and human hepatocytes, (E)-Guggulsterone (1-20 μM) selectively activates ERα (but not ERα subtype) and induces Cyp3a11 and CYP3A4 expression. It inhibits Cu²⁺-mediated LDL lipid peroxidation and oxygen free radical generation (5-20 μM), effectively blocking both enzymatic and non-enzymatic lipid peroxidation processes. In human hepatocellular carcinoma HepG2 cells, guggulsterone (0, 35, 50, 75 μmol/L, 24 hours) alters TGF-β1, TNF-α, and VEGF levels. Guggulsterone (0.5-20 μM, 24 hours) suppresses TREM-1, TLR4 and TNF-α expression as well as the phosphorylation of IκBα and NF-κB p65 induced by LPS.

GugguLsterone (mouse; 100 mg/kg once daily for 8 days) significantly increased the incidence of TNBS-induced cystitis in wild-type mice [2].

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Compared with the control mice or mice treated with a low dose of guggulsterone , high dose of guggulsterone significantly decreased the extent of collagen deposition and the percentage of activated HSCs undergoing apoptosis. Conclusions: These results demonstrate that guggulsterone suppressed HSC activation and survival by inhibiting NF-κB activation and inducing apoptosis. Therefore, guggulsterone may be useful as an antifibrotic agent in chronic liver diseases.[3]

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(E)-Guggulsterone (orally, 100 mg/kg once daily for 8 days) significantly improved the survival rates of wild-type mice with TNBS-induced colitis. It inhibited DSS-induced murine colitis as assessed by reduction in clinical disease activity score, colon length, and histology. Guggulsterone (50 mg/kg po.) significantly protected against cardiac damage in isoproterenol-induced myocardial ischemia in rats, as assessed by reversal of blood and heart biochemical parameters. In an endotoxin-induced uveitis (EIU) rat model, guggulsterone (30 mg/kg, intraperitoneally) prevented increases in infiltrating cells, total protein, MMP-2, NO, and PGE2 in aqueous humor, and also prevented the expression of MMP-2, iNOS, Cox-2, IκB and NF-κB in eye tissues. By inhibiting CDCA-induced transactivation of FXR, (E)-Guggulsterone lowers low-density lipoprotein cholesterol and triglyceride levels in rodents fed a high cholesterol diet.

Triggering receptor expressed on myeloid cells 1 (TREM-1)-expressing intestinal macrophages are significantly increased in the colons of patients with inflammatory bowel disease (IBD). We focused here on the effects of guggulsterone on macrophage modulation in colitis as a potential therapeutic molecule in human IBD and explore the underlying mechanisms. Gene expression in macrophages was examined and wound-healing assay using HT-29 cells was performed. Colitis in wild-type and IL-10-, Toll-like receptor 4 (TLR4)-, and myeloid differentiation primary response 88 (MyD88)-deficient mice was induced via the administration of 2,4,6-trinitrobenzene sulfonic acid (TNBS) into the colon. In both in vitro and in vivo experiments, guggulsterone suppressed intestinal inflammation amplified by TREM-1 stimulation, in which the suppression of NF-κB, activating protein-1, and proteasome pathways was involved. In the TNBS-induced colitis model, guggulsterone reduced disease activity index scores and TREM-1 expression, stimulated IL-10 production, and improved survival in wild-type mice. These effects were not observed in IL-10-, TLR4-, and MyD88-deficient mice. Guggulsterone also suppressed M1 polarization, yet induced the M2 phenotype in macrophages from IBD patients as well as from mice. These findings indicate that guggulsterone blocks the hyperactivation of macrophages via TREM-1 suppression and induces M2 polarization via IL-10 mediated by the TLR4 signaling pathway. Furthermore, this study provides a new rationale for the therapeutic potential of guggulsterone in the treatment of IBD. NEW & NOTEWORTHY We found that guggulsterone attenuates triggering receptor expressed on myeloid cells 1 (TREM-1)-mediated hyperactivation of macrophages and polarizes macrophages toward the M2 phenotype. This was mediated by IL-10 and partly Toll-like receptor 4 signaling pathways. Overall, these data support that guggulsterone as a natural plant sterol modulates macrophage phenotypes in colitis, which may be of novel therapeutic importance in inflammatory bowel disease treatment[2].

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The following experimental protocols are described: FXR antagonism assay – FXR activation induced by chenodeoxycholic acid (CDCA) is measured, and (E)-Guggulsterone decreases CDCA-induced FXR activation with an IC50 of 15 µM. Steroid receptor binding assays – Both (E)- and (Z)-guggulsterone bind to mineralocorticoid receptor (MR) with Ki ≈ 35 nM, and to androgen receptor (AR), glucocorticoid receptor (GR), and progesterone receptor (PR) with Ki values ranging from 224 to 315 nM. In cell-based functional cotransfection assays, guggulsterones behave as antagonists of AR, GR, and MR but as agonists of PR. CYP enzyme inhibition assay – (E)-Guggulsterone inhibits CYP2C9 with IC50 of 19 μM and CYP2C19 with IC50 of 2.1 μM.

The following cell-based assay protocols are described: MCF10A cell Nrf2 activation assay – Human breast epithelial MCF10A cells are treated with (E)-Guggulsterone (5-25 μM, 0-12 hours) to assess Nrf2 signaling pathway activation and HO-1 expression. Huh-7 cell anti-dengue virus assay – Huh-7 cells are treated with (E)-Guggulsterone (0-20 μM, 3 days) to evaluate anti-DENV activity through Nrf2/HO-1 pathway activation. Antibacterial disc diffusion assay – (E)-Guggulsterone (3.2 mM, 24 hours) is tested against Bacillus subtilis, Staphylococcus aureus, and Pseudomonas aeruginosa, with inhibition zone diameters measured. Hepatocyte CYP3A induction assay – Mouse and human hepatocytes are treated with (E)-Guggulsterone (1-20 μM) to assess ERα activation and Cyp3a11/CYP3A4 expression. LDL lipid peroxidation inhibition assay – (E)-Guggulsterone (5-20 μM) is tested for inhibition of Cu²⁺-mediated LDL lipid peroxidation and oxygen free radical generation. HepG2 cell apoptosis assay – Human hepatocellular carcinoma HepG2 cells are treated with guggulsterone (0, 35, 50, 75 μmol/L, 24 hours) and TGF-β1, TNF-α, and VEGF levels are measured by ELISA. LPS-induced inflammation assay – Cells are treated with guggulsterone (0.5-20 μM, 24 hours) to assess suppression of TREM-1, TLR4 and TNF-α expression as well as IκBα and NF-κB p65 phosphorylation.

Thirty-five white male mice of six weeks’ old were used. Their weights ranged between 20-30 gm. The animals were housed in well-ventilated plastic cages, and were maintained under conditions of relatively controlled temperature, humidity, and 12 hrs. light / dark cycle. With a free access to a standard commercial diet that was purchased from the local market and tap water ad libitum. The animals randomly were divided into five groups of 7 animals in each group, as the following: Group I: mice utilized in this group were fed standard commercial diet and considered to be the control group. Group II: mice utilized in this group were fed a specially formulated high-fat diet (HFD) for 12 weeks to induce nonalcoholic liver disease. Group III: mice utilized in this group were fed a high-fat diet that contains Guggulsterone at a concentration of 500 ppm for 12 weeks. Group IV: mice utilized in this group were fed a high-fat diet that contains guggulsterone at a concentration of 1000 ppm for 12 weeks. Group V: mice utilized in this group were fed a high-fat diet that contains Guggulsterone at a concentration of 2000 ppm for 12 weeks. The animal’s weight of all groups was measured routinely once weekly and at zero time. The animals of each group at the end of the experiment after overnight fasting were anesthetized using diethyl ether, and blood was collected from the heart by cardiac puncture, then the animals were sacrificed and the liver were obtained and weighed for calculating liver index (by dividing liver weight in mg by the last body weight. [4]

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Guggulsterone is an active constituent of guggulipid, an ayurvedic drug derived from Commiphora mukul, and is reported to have hypolipidaemic activity. The pharmacokinetics of Z-guggulsterone (1a) and its metabolite, E-guggulsterone (1b), was studied in rats after oral (50 mg kg−1) and intravenous (18 mg kg−1) administration of 1a. It was observed that 1a was isomerized to 1b in treated rat serum samples.[3]
Serum levels of gugulsterone after intravenous administration showed a biexponential elimination phase with a mean ± s.d. terminal half-life of 10.02 ± 4.74 h and 9.24 ± 3.32 h for 1a and 1b isomers, respectively. The values of systemic clearance and AUC for both 1a and 1b were observed to be 0.71 Lh−1, 4.9 μg h mL−1 and 1.04 Lh−1, 3–65 μg h mL−1, respectively. After oral administration, the concentration-time profile declined in a monoexponential fashion with the value of Cmax, terminal half-life, clearance and AUC for 1a and 1b being 1.07 μg ML−1, 4.48 h, 1.76 Lh−1, 5.95 μg mL−1 and 0.97 μ mL−1, 3.56 h, 2.24 Lh−1 and 4.75 μg h mL−1, respectively. Absolute bioavailability of parent compound (1a) after oral administration was 42.9%.[3]

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The following animal experimental protocols are described: Colitis model (TNBS-induced) – Wild-type mice with TNBS-induced colitis are treated orally with (E)-Guggulsterone (100 mg/kg once daily for 8 days), and survival rates are measured. Colitis model (DSS-induced) – DSS-induced murine colitis is assessed by reduction in clinical disease activity score, colon length, and histology following guggulsterone treatment. Myocardial ischemia model – Isoproterenol-induced myocardial ischemia in rats; (E)-Guggulsterone (both E and Z isomers) is administered orally at 50 mg/kg, and cardiac damage protection is assessed by reversal of blood and heart biochemical parameters. Endotoxin-induced uveitis (EIU) model – EIU is induced by subcutaneous injection of lipopolysaccharide (150 μg) into Lewis rats; guggulsterone is administered intraperitoneally at 30 mg/kg body weight. After 24 hours, eyes are enucleated, aqueous humor collected, and infiltrating cells, total protein, MMP-2, NO, PGE2, and inflammatory cytokine expression are measured. Hyperlipidemia model – Rodents fed a high cholesterol diet receive (E)-Guggulsterone; LDL cholesterol and triglyceride levels are measured to assess hypolipidemic effects.

(E)-Guggulsterone is orally active. Both isomers are highly bound to rat plasma proteins (>95% bound). Following oral administration, plasma concentration decreases rapidly, with a terminal half-life of 0.63 ± 0.25 hours for the E-isomer. The clearance for the E-isomer is 2.79 ± 0.73 L/h/kg. In vitro intrinsic clearance (CLint) in rat liver microsomes is 33.34 ± 0.51 μL/min/mg protein for the E-isomer. First-pass metabolism seems to be responsible for low bioavailability of guggulsterone in rats. (E)-Guggulsterone shows high protein binding in human, monkey, rabbit, and rat plasma (>96%) and promotes oxidative metabolism in liver microsomes. ADME prediction shows no violation of Lipinski and Veber rules. The compound may inhibit CYP2C19 and CYP2C9 but cannot inhibit CYP1A2, CYP2D6, and CYP3A4.

(E)-Guggulsterone has several reported side effects, including widespread erythematous papules in a morbilliform pattern, macules localized to the arms, swelling and erythema of the face with burning sensation, pruritis, and bullous lesions on the lower legs with associated headaches, myalgia, and itching. In silico prediction indicates that guggulsterone has no hepatotoxicity, cytotoxicity, or mutagenicity effects. (E)-Guggulsterone induces the expression of human CYP3A. In large quantities, it may lead to damage to health and cause irritation to eyes, skin, or respiratory organs.

[1]. Guggulsterone for Chemoprevention of Cancer. Curr Pharm Des. 2016;22(3):294-306.
[2]. Protective effects of guggulsterone against colitis are associated with the suppression of TREM-1 and modulation of macrophages. Am J Physiol Gastrointest Liver Physiol. 2018 Jul 1;315(1):G128-G139.
[3]. Guggulsterone attenuates activation and survival of hepatic stellate cell by inhibiting nuclear factor kappa B activation and inducing apoptosis. J Gastroenterol Hepatol. 2013 Dec;28(12):1859-68.
[4]. Possible Amelioration of the Severity of Nutritional Steatohepatitis by Guggulsterone in Mice. Iraqi J Pharm Sci, Vol.28(1) 2019. DOI: https://doi.org/10.31351/vol28iss1pp17-23; https://bijps.uobaghdad.edu.iq/index.php/bijps/article/view/801; https://pdfs.semanticscholar.org/fed5/b2b899b009d9a009406450b1d111bb67f233.pdf

E-Glucosterone is a 3-hydroxysteroid with androgenic activity. It has been reported that E-Glucosterone is found in the myrrh tree (Commiphora mukul) and myrrh tree (Commiphora wightii), and relevant data are available for reference.

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(E)-Guggulsterone is a naturally occurring plant sterol (3-hydroxy steroid) found in Commiphora mukul and Commiphora wightii. It is also known as (-)-(E)-Guggulsterone, with molecular formula C21H28O2 and molecular weight 312.45. It potently reverses multi-drug resistance in a number of human cancer cell lines, extending the efficacy of chemotherapy. Guggulsterone is a competitive antagonist of FXR both in vitro and in vivo. It suppresses dengue virus (DENV) replication by upregulating antiviral interferon responses via inducing HO-1 expression through Nrf2 activation. The compound is being investigated for chemoprevention of cancer and has demonstrated activity in head and neck squamous cell carcinoma cell lines with EC50 values ranging from 5 to 8 μM, inducing apoptosis and cell cycle arrest, inhibiting invasion, and enhancing the efficacy of erlotinib, cetuximab, and cisplatin. The cardioprotective and antioxidant activities of synthetic guggulsterone are comparable to those of natural guggulsterone. Guggulsterone and both isomers at concentrations of 5-20 mM inhibit oxidative degradation of lipids in human low-density lipoprotein and rat liver microsomes induced by metal ions. Further studies are needed to evaluate the potential reproductive toxicity of guggulsterone.

C21H28O2

312.44582

312.208

C, 80.73; H, 9.03; O, 10.24

39025-24-6

39025-23-5 (Z-Guggulsterone); 95975-55-6; 39025-24-6 (E-Guggulsterone)

6439929

Typically exists as White to off-white solids at room temperature

1.1±0.1 g/cm3

463.3±45.0 °C at 760 mmHg

170-171.5°

172.3±25.7 °C

0.0±1.1 mmHg at 25°C

1.557

3.65

0

2

0

23

640

5

C/C=C1/C2(C)C(C3C(CC2)C2(C)C(=CC(=O)CC2)CC3)CC/1=O

WDXRGPWQVHZTQJ-AUKWTSKRSA-N

InChI=1S/C21H28O2/c1-4-16-19(23)12-18-15-6-5-13-11-14(22)7-9-20(13,2)17(15)8-10-21(16,18)3/h4,11,15,17-18H,5-10,12H2,1-3H3/b16-4-/t15-,17+,18+,20+,21-/m1/s1

(8R,9S,10R,13S,14S,17E)-17-ethylidene-10,13-dimethyl-1,2,6,7,8,9,11,12,14,15-decahydrocyclopenta[a]phenanthrene-3,16-dione

(E)-Guggulsterone; E-Guggulsterone; 39025-24-6; Guggulsterone E; Guggulsterone; trans-Guggulsterone; Guggulsterones E; Guggulsterone, (E)-;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

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

DMSO : ~25 mg/mL (~80.01 mM)

Solubility in Formulation 1: ≥ 2 mg/mL (6.40 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 20.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)