In MFC-7 cells exposed to 9.37 μM α-ZOL μM, alpha-zearalenol (α-zol) (0.001-10 μM; 24-72 hours) exhibited the highest RPE (90.5%) with an IC50 of 12.5[1]. After 72 hours of treatment, α-Zearalenol (α-zol) (0.001-10 μM; 24-72 hours) mostly increases the viability of MCF-7 cells, but it also slightly damages MDA-MB231 cells [2]. MCF7 cell population is increased in S and G2/M phases by alpha-zearalenol (α-zol) (0.001-10 μM; 24-72 hours); MDA-MB231 cell cycle phase is not found to change. Adaptability [2]. TNF-α expression is lowered by α-zearalenol (α-zol) (1–10 μM; 24 hours). When combined, α-ZOL and β-ZOL have an anti-inflammatory effect on IL-1β and an inhibitory effect on IL-8. High toxin concentrations lead to synergistic effects [2].

Cell Proliferation Assay[1]
Cell Types: MFC-7 Cell
Tested Concentrations: 6.25 µM-25 µM
Incubation Duration: 24 hrs (hours)
Experimental Results: In MFC-7 cells, the highest RPE (90.5%) was observed at 9.37 µM α-ZOL exposure .

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Cell viability assay [2]
Cell Types: Breast Cancer
Cell Types: MCF-7 and MDA-MB231
Tested Concentrations: 0.001 μM, 0.1 μM and 10 μM
Incubation Duration: 24 hrs (hours), 48 hrs (hours), 72 hrs (hours)
Experimental Results: Changes in both cell types Growing breast cancer cell lines.

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Cell cycle analysis [2]
Cell Types: MCF7 cells, MDA-MB231 cells
Tested Concentrations: 0.001 μM, 0.1 μM and 10 μM
Incubation Duration: 72 hrs (hours)
Experimental Results: An increase in cell cycle was observed in MCF7 cells.

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RT-PCR[2]
Cell Types: HepG2 Cell
Tested Concentrations: 1 μM, 5 μM, 10 μM
Incubation Duration: 72 hrs (hours)
Experimental Results: Induced TNF-α, IL-1β and IL-8 expression diminished, that is, in most cases, lower than High doses were statistically significant.

Toxicity Summary
Mycotoxins, such as α-zearalenol (α-ZOL) and β-zearalenol (β-ZOL), as feed contaminants, can impair livestock fertility and cause fetal developmental abnormalities. Adding α- or β-ZOL (7.5, 15, and 30 μM) to cultures stimulated with follicle-stimulating hormone (FSH, 0.01 μg) or trichomonadine (10 μM) reduced progesterone synthesis and the levels of p450scc and 3β-HSD transcripts in a dose-dependent manner (P<0.05). These mycotoxins also reduced the enzyme activity of 3β-HSD and the abundance of the p450scc protein. The effects of mycotoxins on FSH receptor-dependent and receptor-independent pathways suggest that adenylate cyclase activity and/or downstream regulatory pathways are targets of mycotoxin action. A significant dose-dependent decrease in p450scc and 3β-HSD transcripts suggests that α- and β-zearalenones affect the transcriptional regulation of these enzymes. Detection of α- and β-zearalenone derivatives metabolized in the liver (as excretion products) showed that the β-epimer is dominant in pigs and humans. (A15419) Typically, α-zearalenone has approximately 50% of the estrogenic activity of E2, and its estrogenic activity order (both in terms of in vitro receptor competitive binding and in vivo induction of Vtg and Zr protein levels) is: α-zearalenone > β-zearalenone. Studies have also found that the fungal toxins α-zearalenol and β-zearalenol affect apoptosis and proliferation in equine ovarian granulosa cells. (L2099) The mechanisms by which α- or β-zearalenol mediates their cytotoxic effects appear to vary depending on cell type and the toxins exposed. In evaluating the toxicity of α-zearalenone and β-zearalenone to RAW264.7 macrophages, α-zearalenone not only reduced cell viability more significantly than β-zearalenone, but also induced cell death primarily through apoptosis rather than necrosis. Zearalenone metabolites induced loss of mitochondrial membrane potential (MMP), alterations in mitochondrial Bcl-2 and Bax proteins, and cytoplasmic release of cytochrome c and apoptosis-inducing factor (AIF). Use of specific inhibitors targeting c-Jun N-terminal kinase (JNK), p38 kinase, or p53 (rather than pan-caspase or caspase-8 inhibitors) reduced toxin-induced reactive oxygen species (ROS) production and attenuated the cell viability decrease induced by α-zoledronic acid (ZOL) or β-zoledronic acid (ZOL). Zearalenone metabolites activate p53, JNK, or p38 kinases, which are key upstream signals required for altered mitochondrial Bcl-2/Bax signaling pathways and intracellular ROS generation. Meanwhile, loss of mitochondrial membrane potential (MMP) and AIF nuclear translocation are key downstream events mediated by zearalenone metabolites in macrophage apoptosis. (A15420)

[1]. Estrogenic activity of zearalenone, α-zearalenol and β-zearalenol assessed using the E-screen assay in MCF-7 cells. Toxicol Mech Methods. 2018 May;28(4):239-242.

[2]. Cytotoxic and inflammatory effects of individual and combined exposure of HepG2 cells to zearalenone and its metabolites. Naunyn Schmiedebergs Arch Pharmacol. 2019 Aug;392(8):937-947.

[3]. Effects of α-zearalenol on the metabolome of two breast cancer cell lines by 1H-NMR approach. Metabolomics. 2018 Feb 14;14(3):33.

α-Zearabenol is a macrocyclic lactone compound. α-Zearabenol belongs to the class of macrocyclic lactones and their analogues. These compounds are organic compounds containing at least twelve lactone rings. The term "macrocyclic lactone" encompasses a series of unrelated compounds with large macrocyclic lactam rings. See also: Zearabenol (note moved here).

C₁₈H₂₄O₅

320.38

320.162

36455-72-8

5284645

White to off-white solid powder

1.2±0.1 g/cm3

599.0±50.0 °C at 760 mmHg

158-161°C

217.9±23.6 °C

0.0±1.8 mmHg at 25°C

1.549

4.17

3

5

0

23

408

2

C[C@H]1CCC[C@@H](CCC/C=C/C2=C(C(=CC(=C2)O)O)C(=O)O1)O

FPQFYIAXQDXNOR-QDKLYSGJSA-N

InChI=1S/C18H24O5/c1-12-6-5-9-14(19)8-4-2-3-7-13-10-15(20)11-16(21)17(13)18(22)23-12/h3,7,10-12,14,19-21H,2,4-6,8-9H2,1H3/b7-3+/t12-,14+/m0/s1

(4S,8R,12E)-8,16,18-trihydroxy-4-methyl-3-oxabicyclo[12.4.0]octadeca-1(14),12,15,17-tetraen-2-one

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 : ~100 mg/mL (~312.13 mM)

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.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*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).

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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)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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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).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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