PI3K; Akt
PI3K (phosphatidylinositol 3-kinase) [3]
Akt [3]
IKK (IκBα kinase) [3]
NF-κB [3]
AMPK (AMP-activated protein kinase) [3]
mTOR [3]
survivin [3]
Hsp90α (heat-shock protein 90α) - Deguelin specifically binds to the ATP pocket of Hsp90α [3]
HIF-1α [3]
VEGF [3]
Cyclin E [3]
pRb [3]
E2F1 [3]

Deguelin downregulates Akt phosphorylation in leukaemia cell lines with an active PI3K/Akt axis. Deguelin is effective at preventing Akt phosphorylation at concentrations of 10 or 100 nmol/l. By deguelin, total Akt expression is unaffected. Deguelin also has no impact on the phosphorylation or expression of the p44/42 or p38 MAP kinases in U937 cells. Deguelin makes human leukemia cells more susceptible to chemotherapy. Deguelin increases cytarabine sensitivity and dephosphorylates Akt in AML blasts, but not in CB CD34+. When used for 24 hours at a concentration of 10 nmol/l, deguelin arrests U937 cells in the S phase and prevents them from progressing to the G2/M phase. Deguelin does not significantly increase the apoptotic rate of U937 cells when used alone for 24 hours at a concentration of 10 nmol/l[1].

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Deguelin induced apoptosis in premalignant and malignant human bronchial epithelial (HBE) cells at 10⁻⁷ M, showing morphological changes typical of apoptosis, with no effect on healthy HBE cells. It inhibited PI3K activity and downregulated pAkt level without affecting MAPK pathways [3].
Deguelin induced apoptosis by inhibiting the PI3K/Akt pathway in human leukemia HL-60, U937 and human breast cancer MCF-7 cells [3].
Deguelin treatment reduced pAkt level, inhibiting pro-apoptotic proteins Bad and Bax by phosphorylation, and activating caspase-3 [3].
Deguelin suppressed NF-κB activation by inhibiting IKK activation, protecting IκBα from degradation, thereby masking nuclear localization signals of NF-κB and keeping IκBα outside the nucleus [3]. It also suppressed expression of NF-κB downstream anti-apoptotic proteins including Bcl-2, Bcl-xl, Bfl-1, IAP1/2 and TRAF-1 [3].
Deguelin blocked NF-κB activation induced by TNFR1, TRAF2, TRADD and NIK, but not by p65, indicating it acts upstream of p65 in the NF-κB pathway [3].
In B-chronic lymphocytic leukemia (B-CLL) cells, Deguelin promoted IκBα degradation [3]. In leukemia cells, Deguelin inhibited IκBα expression and induced apoptosis via NF-κB pathway [3].
Deguelin activated AMPK by ATP depletion and inhibited Akt activation in HBE cells; suppressed Akt promoted AMPK function, activating TSC2, which inhibited mTOR, p70S6, and activated 4E-BP1, ultimately suppressing survivin expression [3].
Deguelin increased p27 expression, leading to cyclin E inactivation, pRb dephosphorylation, and hypophosphorylated pRb binding to E2F1, causing G1-S phase cell cycle arrest in colon cancer cells [3].
In pre-malignant HBE cells, Deguelin treatment increased expression of p21 and p27 in a dose-dependent manner [3].
Deguelin induced cyclin D1/pRb decrease and subsequent G1-S phase arrest in breast cancer MDA-MB-231 and lymphoma Daudi cells [3].
Deguelin (5 nmol) significantly reduced the number of endothelial cell sprouts in chick aortic arch ring assay. In chick chorioallantoic membrane assay, Deguelin (1 nmol/egg) suppressed new vessel formation to 46.7±3.33% (n=20) [3].
Deguelin (100 nmol) arrested HIF-1α protein synthesis and initiated ubiquitin-mediated protein degradation, resulting in downregulation of VEGF and bFGF. HIF-1α degradation was correlated with Hsp90 binding [3].

Deguelin inhibits in vivo angiogenesis of chick chorioallantoic membrane (CAM) without cytotoxic effect and significantly reduces laser-induced CNV in a mouse model of AMD without significant retinal toxicity[2]. It demonstrated notable in vitro and in vivo anti-tumorigenesis and anti-proliferative activity in different cancer types. Deguelin significantly decreased the incidence of tumors in pre-clinical studies. Deguelin, when applied topically, significantly reduced the number of skin tumors that were caused by UVB exposure, suggesting that it may have potential as a cancer chemopreventive. In A/J mice exposed to the tobacco-specific carcinogen benzo(a)pyrene (Bap) and other carcinogens, deguelin significantly reduced tumor multiplicity and volume as well as the overall tumor burden with no discernible toxicity. Deguelin is toxic above a certain dose, though, so this should not be disregarded. Parkinson's disease was brought on by the treatment with deguelin, a potential mitochondria complex I inhibitor, which reduced tyrosine hydroxylase-positive neurons. Kim et al shows that deguelin promoted a PD-like syndrome, mainly by Src/STAT signaling, since α-synuclein (a key protein function in the pathogenesis of PD) was phosphorylated by deguelin-activated Src[3].
In early animal experiments (1997), Deguelin at 33 µg/rat decreased tumor incidence from 60% (control) to 10%, and tumor multiplicity from 4.2 to 0.1; at 330 µg/rat, no tumors were observed [3].
Topically-administered Deguelin significantly suppressed the multiplicity of UVB-induced skin tumors [3].
In A/J mice, Deguelin reduced tumor multiplicity, volume, and overall tumor burden after exposure to tobacco-specific carcinogen benzo(a)pyrene (BaP) and other carcinogens, with no detectable toxicity [3].
Deguelin (4 mg/kg) significantly inhibited growth of H1299 xenograft tumors (treatment group tumor volume 115.9 mm³ vs. control 798 mm³) without detectable toxic effects [3].
Micro-vessel density in xenograft tumors from Deguelin-treated mice was decreased, reflected by tumor vessel number/high-power field from 100 to 58% [3].

Caspase 3 activity is determined using Caspase-Glo-3 assays. This assay provides luminogenic substrate in a buffer system optimized for each specific caspase activity. The caspase cleavage of the substrate is followed by generation of a luminescent signal. The signal generated is proportional to the amount of caspase activity present in the sample. Protein (10 µg) from the cell samples is diluted in water to a final volume of 50 µL, then 50 µL of Caspase-Glo-3 reagent is added to a white 96-well microtitre plate. The plate is sealed, gently mixed for 30 seconds at 300-500 rpm, and then incubated for 30 minutes at room temperature. In a microplate reader (TECAN Infinite 200), luminescence is measured.

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Not described in this review article.

Deguelin is incubated with breast cancer cells at increasing concentrations for 24, 48, and 72 hours. The concentrations range from 31 nM to 500 nM. At the conclusion, cells are trypsinized, and cell proliferation is assessed by counting cells using a Z-series Coulter counter. Data are displayed as MeanSE% of control.

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For cell cycle and apoptosis analysis: Cells were treated with deguelin at indicated concentrations for 24 h, then harvested, fixed with 70% cold ethanol for 1 h, stained with propidium iodide, and analyzed by flow cytometry for subdiploid DNA content. Alternatively, cells were stained with Annexin V-FITC in binding buffer and analyzed by flow cytometry after addition of propidium iodide to discriminate live, apoptotic, and necrotic cells. [1]
For Western blot analysis: Cells were lysed in buffer containing Tris-HCl, MgCl2, EGTA, Triton X-100, sucrose, protease inhibitor cocktail, NaF, 2-glycerophosphate, and NaPPi. After homogenization and centrifugation, protein concentration was determined. Protein (80 μg per lane) was separated by SDS-PAGE, transferred to nitrocellulose membranes, blocked with blocking buffer (PBS containing normal goat serum, BSA, non-fat dried milk), incubated overnight with primary antibodies against total Akt, Thr308 p-Akt, Ser473 p-Akt, total p44/42 MAP kinase, Thr202/Tyr204 p-MAP kinase, total p38 MAP kinase, Thr180/Tyr182 p-p38 MAP kinase, or β-tubulin, then incubated with peroxidase-conjugated secondary antibody, and visualized by enhanced chemiluminescence. [1]
For isolation and culture of human cord blood CD34+ cells: Cord blood mononuclear cells were isolated by density gradient centrifugation, adherent cells removed, and CD34+ cells selected using magnetic cell sorting. Purity (93-98%) was determined by flow cytometry. Cells were cultured in serum-free medium supplemented with nucleosides, BSA, insulin, iron-saturated transferrin, 2-mercaptoethanol, IL-3 (5 ng/mL), SCF (50 ng/mL), and IL-6 (10 ng/mL). For erythroid differentiation, cells were incubated for 6 days with EPO (5 U/mL) and SCF (50 ng/mL) without IL-3 and IL-6, then double-stained with anti-CD71-FITC and anti-glycophorin A-PE and analyzed by flow cytometry. [1]
For AML blast culture: AML blasts (>80% purity) were cultured in methylcellulose medium supplemented with IL-3 (20 ng/mL), IL-6 (20 ng/mL), and SCF (50 ng/mL). [1]
For MTT cell viability assay (HUVECs): HUVECs (1×10^5 cells/well) were plated in 96-well plates, cultured overnight, treated with deguelin (0.01-10 μM) for 48 h, then medium replaced with fresh medium containing 0.5 mg/mL MTT for 4 h. Formazan was solubilized with DMSO and absorbance measured at 540 nm. [2]
For tube formation assay: HUVECs (1×10^5 cells) were seeded on Matrigel-coated surfaces and treated with 0.1 μM deguelin or VEGF (20 ng/mL) for 18 h. Tube formation was quantified by counting connected cells in randomly selected fields at 200× magnification. [2]
For TUNEL assay (retinal toxicity): Mice were intravitreally injected with 1 μM deguelin, sacrificed after 3 days, eyes enucleated, fixed in paraformaldehyde, embedded in paraffin, and TUNEL staining performed using a fluorescence kit. TUNEL-positive cells were evaluated under fluorescence microscopy. [2]

Female athymic mice (nu/nu), which are six to seven weeks old, are kept in barrier-free housing with 12-hour light/12-hour dark cycles in an environment with a temperature of 24°C and a relative humidity of 50%. Water is available at all times, along with sterile mouse food. The dorsal flank region of the animal is subcutaneously injected with a 23 g hypodermic needle after MDA-MB-231 cells (3.0 million cells/animal) are injected in sterile PBS suspension. A palpable tumor at the injection site is checked daily on the animals. The mice are divided randomly into three groups and given one of the following after the tumor (about 50 mm3) appears: 1) a vehicle as a control. 2) Deguelin therapy at a dose of 2 mg/kg body weight; or 3) Deguelin at a dose of 4 mg/kg body weight. Ten creatures make up each group. For a period of 21 days, vehicle or deguelin is given by i.p. injection. The toxicity of the drugs and vehicles is observed in animals every day, and they are weighed once per week. Every other day, the tumor's growth is observed, and every other day, calipers are used to measure the tumor's size. The widely used equation tumor volume (mm3)=π/6 lengthwidthdepth is used to calculate tumor volume. The information shown in each group's average tumor volume plus SE (mm3). Except when they appear to be moribund or tumors exhibit necrosis, the animals are sacrificed at the appointed time. At termination, the tumor is excised, freed from connective tissue and other organs, a small piece is fixed in 10% buffered formalin, and the remaining tumor is snap-frozen for a future biochemical analysis. The liver, lung, kidney, and spleen are removed and weighed.

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Eight-week-old male and female C57BL/6 mice were used. The experimental procedures were approved by the Animal Ethics Committee of Southern Medical University. The mice were housed in a controlled environment with 12 h light/dark cycles, a temperature of 25 ± 2°C, humidity between 40% between 60%, and had free access to water and food. Referring to our previous study,11 the ALF model was induced by APAP (300 mg/kg, Macklin, Shanghai, China) via oral gavage at 8:00 PM. To investigate the protective role of deguelin (dissolved in sesame oil) in ALF, the APAP-treated mice were immediately injected intraperitoneally with deguelin (20 mg/kg, InvivoChem, Guangzhou, China) or sesame oil for 24 h. For inhibitor experiment, ML224 (10 mg/kg, InvivoChem, Guangzhou, China) was injected intraperitoneally into mice. Reference: Gut Microbes. 2024 Jan-Dec;16(1):2404138.

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For laser-induced CNV in mice: Seven- to eight-week-old female C57BL/6J mice were anesthetized, pupils dilated, and three burns of 831-nm diode laser photocoagulation (75-μm spot size, 0.1-s duration, 120 mW) were delivered to each 3, 6, 9, and 12 o'clock position approximately two disc diameters from the optic disc using a photocoagulator with a handheld +78 diopter lens. Bubble or pop sensation indicated successful rupture of Bruch's membrane. On day 10 after laser photocoagulation (when maximal CNV began), mice were intravenously injected with 0.1 μM deguelin in 1 μL phosphate-buffered saline. At day 14, mice were perfused through the tail vein with high-molecular-weight (500,000) fluorescein-conjugated dextran dissolved in PBS. After 1 h perfusion, eyes were enucleated, fixed in 4% paraformaldehyde for 4 h, then flat-mounted or embedded in paraffin. Sagittal sections (4-5 μm thick, each 10 μm apart) were cut through the center of the laser photocoagulation site, stained with hematoxylin and eosin, and vessels from subretinal fibrovascular membrane were counted in five sections per site by two independent observers blinded to treatment. At least 25 animals per group and 100 laser sites were evaluated. [2]
For retinal toxicity assessment: Deguelin (1 μM) was intravitreally injected into mice. After 3 days, eyes were enucleated, fixed, paraffin-embedded, sectioned, and examined by light microscopy for histological changes. TUNEL staining was also performed to detect apoptotic cells. [2]

Oral LD50 in rats >2 g/kg

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Deguelin is a potential mitochondria complex I inhibitor [3].
Long-term and high-dose Deguelin administration reduced tyrosine hydroxylase-positive neurons, leading to Parkinson's disease (PD)-like syndrome; promoted PD-like syndrome mainly by Src/STAT signaling, as α-synuclein was phosphorylated by deguelin-activated Src [3].
No sign of overt toxicity was observed at the dose of 2-4 mg/kg [3].
At doses of 33 µg/rat and 330 µg/rat, no toxicity was reported in early animal experiments [3].
However, over a certain dose, toxicity should not be neglected; deguelin is closely correlated with the pathogenesis of Parkinson's disease [3].

[1]. Br J Haematol. 2005 Jun;129(5):677-86.

[2]. J Pharmacol Exp Ther. 2008 Feb;324(2):643-7.

[3]. Mol Clin Oncol. 2013 Mar;1(2):215-219.

Rotenones, chemically named 13,13a-dihydro-3H-chromeno[3,4-b]pyrano[2,3-h]chromeno-7(7aH)-one, are substituted with methoxy groups at positions 9 and 10 and with two methyl groups at position 3 (7aS,13aS-stereoisomers). They are abundant in the bark, roots, and leaves of leguminous plants and have been reported to have antitumor effects against various cancers. They possess a variety of activities, including inducing apoptosis, antitumor activity, acting as a plant metabolite, inhibiting angiogenesis, antiviral activity, inhibiting mitochondrial NADH:ubiquinone reductase, anti-inflammatory activity, and inhibiting EC 2.7.11.1 (non-specific serine/threonine protein kinase). They belong to the rotenone class of compounds and are aromatic ethers, organic heteropentane compounds, and diethers. Houttuynin has been reported in fish species with relevant data (such as Houttuynia cordata and Houttuynia elliptica).

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Deguelin is a natural rotenoid with anti-tumorigenic, anti-proliferative and anti-angiogenic activities. It acts as a special inhibitor of Hsp90 chaperone function, leading to decreased expression of oncogenic proteins including MEK1/2, Akt, HIF-1α, COX-2 and NF-κB [3].
Derivatives such as SH-14 (via hydroxylation of 7α carbon or functional group substitution of 7 carbon) showed highest apoptotic activity with no detectable effect on PD evocation [3].
Deguelin is superior for various cancer cells: Colo 16 and SRB-12 skin cancer cells, normal HBE and BEAS-2B HBE cells, MKN-28 and SNU-484 gastric cancer cells [3].
Compared to 17-AAG (an Hsp90 inhibitor in phase II clinical trials with various side-effects), Deguelin demonstrated antitumor activity at a certain dose with no toxicity to healthy cells [3].

C23H22O6

394.4172

394.141

C, 70.04; H, 5.62; O, 24.34

522-17-8

522-17-8

107935

Light yellow to yellow solid powder

1.3±0.1 g/cm3

560.1±50.0 °C at 760 mmHg

85-87ºC(lit.)

244.7±30.2 °C

0.0±1.5 mmHg at 25°C

1.584

5.03

0

6

2

29

674

2

O1C2C3C([H])=C([H])C(C([H])([H])[H])(C([H])([H])[H])OC=3C([H])=C([H])C=2C([C@@]2([H])C3=C([H])C(=C(C([H])=C3OC([H])([H])[C@@]12[H])OC([H])([H])[H])OC([H])([H])[H])=O

ORDAZKGHSNRHTD-UXHICEINSA-N

InChI=1S/C23H22O6/c1-23(2)8-7-12-15(29-23)6-5-13-21(24)20-14-9-17(25-3)18(26-4)10-16(14)27-11-19(20)28-22(12)13/h5-10,19-20H,11H2,1-4H3/t19-,20+/m1/s1

(7aS,13aS)-9,10-dimethoxy-3,3-dimethyl-13,13a-dihydro-3H-pyrano[2,3-c:6,5-f'''']dichromen-7(7aH)-one.

(-)-Deguelin; (-)-cis-deguelin; DEGUELIN(-); CHEBI:4357; K5Z93K66IE; MFCD01740600; (-)-cis-Deguelin

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: ~78 mg/mL (197.8 mM)
Water: <1 mg/mL
Ethanol: 78 mg/mL (197.8 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.34 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.

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Solubility in Formulation 2: 2.5 mg/mL (6.34 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.
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.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (6.34 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.

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