PI3K; Akt

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].

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].

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.

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.

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.

Rat LD oral >2 gm/kg

[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.

Deguelin is a rotenone that is 13,13a-dihydro-3H-chromeno[3,4-b]pyrano[2,3-h]chromen-7(7aH)-one substituted by methoxy groups at positions 9 and 10, and by two methyl groups at position 3 (the 7aS,13aS-stereoisomer). It exists in abundant quantities in the bark, roots, and leaves of the Leguminosae family of plants and reported to exert anti-tumour effects in various cancers. It has a role as an apoptosis inducer, an antineoplastic agent, a plant metabolite, an angiogenesis inhibitor, an antiviral agent, a mitochondrial NADH:ubiquinone reductase inhibitor, an anti-inflammatory agent and an EC 2.7.11.1 (non-specific serine/threonine protein kinase) inhibitor. It is a member of rotenones, an aromatic ether, an organic heteropentacyclic compound and a diether.
Deguelin has been reported in Piscidia piscipula, Derris elliptica, and other organisms with data available.

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.)