Ergosterol is the primary sterol in fungi and is essential for maintaining fungal cell membrane integrity and fluidity. Its biosynthesis is a target for antifungal agents, notably azoles, which inhibit ergosterol synthesis. Ergosterol also exhibits multifaceted bioactivities, including anti-inflammatory, antibacterial, antiviral, and anticancer properties. It inhibits NO production with IC₅₀ values of 27,900 nM and 30,600 nM in different inflammatory models.

Ergosterol is a sterol that was isolated from Grifola frondosa that has applications in the research of allergy disorders that depend on mast cells. Ergosterol (10, 20, and 50 μM) suppresses the release of histamine and beta-hexosaminidase in response to antigen stimulation in RBL-2H3 cells. The levels of TNF-α and IL-4 mRNA were dramatically decreased by ergosterol (20 and 50 μM). FcεRI aggregation generated by antigen is inhibited by ergosterol (50 μM) [1].
In vitro, ergosterol exhibits anti-inflammatory, antibacterial, antiviral, and anticancer properties. It inhibits NO production with IC₅₀ values of 27,900 nM (NOX inhibition in fMLP-induced PMN cells) and 30,600 nM (iNOS inhibition in mouse BV2 microglial cells). It demonstrates antibacterial activity against Legionella pneumophila (MIC: 250 µg/mL) and inhibits the growth of Pseudomonas aeruginosa and Staphylococcus aureus. It shows cytotoxicity against various cell lines, including mouse L5178Y (EC₅₀: 12,600 nM), human COLO 205 (IC₅₀: 4,900 nM), and COLO 320 (IC₅₀: 6,500 nM).

Ergosterol (25, 50 mg/kg, oral) enhanced SOD activity, decreased the production of MDA, CK-MB, and LDH in LPS-induced septic rats, and considerably mitigated the LPS-induced decline in cardiac function in rats [2].
In vivo, ergosterol demonstrates significant antitumor activity in xenografted mouse models with Lewis lung cancer cells, with 100% growth inhibition at 12.5 µg/mL after 48 hours and evidence of metastasis inhibition or tumor regression. Despite some toxicity-related net weight reduction, the compound maintains 100% survival rates in treated mouse models. It also inhibits TNFα-induced NF-κB activity in HeLa cells with an IC₅₀ of 0.91 µg/mL.

Non-cellular enzyme assays for ergosterol involve assessing its antioxidant activity using cell-free systems. The compound's ability to scavenge free radicals can be evaluated using DPPH (2,2-diphenyl-1-picrylhydrazyl) or ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) radical scavenging assays. Its ability to inhibit lipid peroxidation can be measured using the thiobarbituric acid reactive substances (TBARS) assay. Its inhibition of NO production can be assessed using Griess reagent assays in cell-free systems.

In vitro cellular assays for ergosterol involve treating immune cells (e.g., macrophages, microglia) or cancer cell lines with the compound and assessing cell viability, inflammatory responses, and anticancer activity. Cell viability is measured using MTT or CellTiter-Glo® assays. NO production is measured using the Griess reagent. Cytokine production is measured by ELISA. Apoptosis is assessed by flow cytometry using Annexin V/PI staining or by measuring caspase activity.

In vivo animal experiments with ergosterol are conducted in mouse xenograft models of cancer. Tumor-bearing mice are treated with ergosterol via oral or intraperitoneal administration, and tumor growth inhibition is monitored. In Lewis lung cancer models, the compound shows 100% growth inhibition at 12.5 µg/mL after 48 hours. In inflammatory models, the compound's effects on cytokine production and tissue inflammation are assessed. Survival rates and toxicity are also monitored.

Absorption, Distribution and Excretion
A small amount of vitamin D is secreted into breast milk. There is a direct relationship between maternal serum vitamin D levels and the concentration of vitamin D in breast milk. Long-term maternal intake of high doses of vitamin D may lead to higher than normal vitamin D activity in breast milk, resulting in hypercalcemia in infants. /Vitamin D/ Experiments on rats fed a diet containing ergosterol for an extended period showed that only trace amounts of ergosterol were present in liver tissue, lower than the ergosterol concentration that produced an effect in the model experiment. Ergosterol was not detected in the liver after 3 days of the experiment. Simultaneously, the study found that approximately 16% of the oral intake of ergosterol was unmetabolized in rat feces. Possible ergosterol metabolites (dehydroneoergosterol-24-methyl-1,3,5(10),6,8(9),22-hexen-3β-ol; 24-methylcholest-7,24(28)-dien-3β-ol; 4-cholest-7,22,25(?)-trien-3β-ol; 4-methylcholest-7,22(?)-dien-3β-ol, etc.) were identified in feces. Metabolites/Metabolites… The metabolism of ergosterol by cytochrome P450scc was confirmed in both the recombinant system and isolated adrenal mitochondria. The major reaction product was identified as 17α,24-dihydroxyergosterol. Purified P450scc also produced a small amount of hydroxyergosterol. This product is likely an intermediate in the synthesis of 17α,24-dihydroxyergosterol. Unlike cholesterol and 7-dehydrocholesterol, no cleavage of the ergosterol side chain was observed. NMR analysis clearly located a hydroxyl group at C24, and there was evidence that a second hydroxyl group was located at C17. 17α,24-dihydroxyergosterol inhibited the proliferation of HaCaT keratinocytes and melanoma cells. Therefore, compared with cholesterol and 7-dehydrocholesterol, the 24-methyl and C22-C23 double bond of ergosterol prevented the cleavage of the P450scc side chain and transferred the hydroxylase activity of the enzyme from the C22 and C20 sites to the C24 and C17 sites, thereby generating the bioactive product. To investigate the effects of different C-24 stereochemistry on the oxidation of sterols at the C-26 site in microorganisms, ergosterol was transformed using a genetically modified Mycobacterium tuberculosis strain BCS 396. Ergosterol was converted into methyl 3-oxo-4,22-ergosteradien-26-olate, methyl 3-oxo-1,4,22-ergostertrien-26-olate, and 3-oxo-1,4,22-ergostertrien-26-olate, the structures of which were determined by infrared spectroscopy, proton NMR spectroscopy, carbon NMR spectroscopy, and mass spectrometry. The X-ray crystal structure of methyl 3-oxo-4,22-ergosteradien-26-olate showed that the oxidation at the C-26 position of the ergosterane side chain, induced by the chiral center at the C-24 position, resulted in the formation of an S-configuration chiral center at the C-25 position. Long-term feeding of rats with ergosterol-containing diets showed that only trace amounts of ergosterol were present in the liver tissue, at concentrations far lower than the effective ergosterol concentrations observed in the model experiments. After 3 days of experimentation, ergosterol was not detected in the liver. Meanwhile, the study found that approximately 16% of the orally administered ergosterol was unmetabolized in rat feces. Furthermore, possible metabolites of ergosterol were identified, such as dehydroneoergosterol-24-methyl-1,3,5(10),6,8(9),22-hexen-3β-ol; 24-methylcholest-7,24(28)-dien-3β-ol; 4-cholest-7,22,25(?)-trien-3β-ol; 4-methylcholest-7,22(?)-dien-3β-ol, etc. Feces.
The metabolism of orally administered ergosterol (Erg) in rats and the bioactivity of vitamin D were investigated. Most of the orally administered Erg was excreted in feces, with the remaining sterols absorbed through the intestines. The absorbed sterols were not transported intact through the skin but were metabolized into campesterol and cholesterol within 25 hours, respectively. No increase in intestinal calcium absorption or plasma calcium concentration was observed in vitamin D-deficient rats after oral administration of Erg. The effects of dietary ergosterol on the responses to oral progesterone and estrogen were investigated using a chicken oviduct assay. Progesterone alone had no effect on the oviducts, but estrogen-induced oviduct hypertrophy was significantly enhanced with concurrent progesterone treatment at all tested dose levels. Ergosterol had no effect on any response of the oviducts studied.
Ergosterol has a molecular weight of 396.65 and a molecular formula of C₂₈H₄₄O. It is a solid powder with a purity of ≥98%. The compound is typically stored at -20°C, protected from light and moisture. It is soluble in organic solvents such as ethanol, DMSO, and chloroform. The compound is stable under recommended storage conditions.

Interactions
Mouse leukemia cells (L1210) cultured in a medium containing ergosterol (40 μg/ml) showed higher antibiotic sensitivity to the control group after transient exposure to amphotericin B (0-10 μg/ml). After 4 hours of culture in the presence of 5 × 10⁻⁹ mol ketoconazole, the acetate incorporation activity of Candida albicans was inhibited by approximately 50%. The inhibitory effect of the antifungal drug miconazole nitrate on ergosterol biosynthesis in Candida albicans was investigated after the addition of the antifungal drug. In vitro drug exposure was performed at 1, 4, 16, and 24 hours. The function and biosynthesis of sterols have long been effective targets for fungal control in various fields, including pharmaceutical and agricultural applications. Fungi are among the organisms that synthesize sterols, primarily ergosterol. This study investigated the effect of dibutyryl cyclic adenosine monophosphate (db-cAMP) on ergosterol levels and the interaction between drugs that can alter cAMP concentrations and antifungal agents. Sterols were extracted from Candida albicans, and ergosterol content was determined by gas chromatography. Interactions between different drugs were determined using the broth dilution method. The study found that phosphodiesterase inhibitors could reverse the inhibitory activity of azole antifungal drugs. Ergosterol levels in Candida albicans incubated with db-cAMP were assessed, and the results showed that db-cAMP increased ergosterol levels. Further experiments provided evidence that the interaction between azole drugs and phosphodiesterase inhibitors is attributable to the relationship between ergosterol and cAMP. Potential implications of this interaction include enhancing the antifungal activity of drugs by modulating cAMP levels. For more complete data on ergosterol interactions (6 entries total), please visit the HSDB record page.
Ergosterol is generally considered to have low toxicity at therapeutic doses. In preclinical studies, it has been generally well-tolerated, although some toxicity-related net weight reduction has been observed at higher doses. Standard laboratory safety precautions should be followed when handling the compound. It may cause skin, eye, and respiratory irritation. Appropriate personal protective equipment should be used.

[1]. Kawai J, et al. Ergosterol and its derivatives from Grifola frondosa inhibit antigen-induced degranulation of RBL-2H3 cells by suppressing the aggregation of high affinity IgE receptors. Biosci Biotechnol Biochem. 2018 Jul 2:1-9.
[2]. Xu J, et al. Ergosterol Attenuates LPS-Induced Myocardial Injury by Modulating Oxidative Stress and Apoptosis in Rats. Cell Physiol Biochem. 2018;48(2):583-592

Therapeutic Uses
This treatment involves the use of very high doses of vitamin D during pregnancy to treat hypoparathyroidism in pregnant women. In two studies, 15 pregnant women received an average of 107,000 IU of vitamin D daily throughout their pregnancy to maintain normal serum calcium levels. All 27 children were born and followed up (up to 16 years old) with normal development. /Vitamin D/ …Irradiated ergosterol from yeast became a major source of vitamin D for food fortification and the treatment of rickets, sparking a public health campaign aimed at eradicating rickets by the 1930s. /Irradiated Ergosterol/

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Drug Warnings Vitamin D is excreted in small amounts into breast milk. There is a direct relationship between maternal serum vitamin D levels and vitamin D concentrations in breast milk. Long-term high maternal intake of vitamin D may lead to excessively high vitamin D activity in breast milk, resulting in hypercalcemia in infants.
Ergosterol (CAS 57-87-4) is the primary sterol in fungi and a provitamin for vitamin D2. It exhibits anti-inflammatory, antibacterial, antiviral, and anticancer properties. Ergosterol inhibits NO production with IC₅₀ values of 27.9 µM and 30.6 µM. It demonstrates antitumor activity in mouse xenograft models, with 100% growth inhibition at 12.5 µg/mL. The compound is used as a research tool for studying fungal biology, antifungal drug development, and cancer research.

C28H44O

396.65

396.339

57-87-4

444679

White to yellow solid powder

1.0±0.1 g/cm3

501.5±39.0 °C at 760 mmHg

156-158 °C(lit.)

216.3±19.3 °C

0.0±2.9 mmHg at 25°C

1.543

9.3

1

1

4

29

712

8

C[C@H](/C=C/[C@H](C)C(C)C)[C@H]1CC[C@@H]2[C@@]1(CC[C@H]3C2=CC=C4[C@@]3(CC[C@@H](C4)O)C)C

DNVPQKQSNYMLRS-APGDWVJJSA-N

InChI=1S/C28H44O/c1-18(2)19(3)7-8-20(4)24-11-12-25-23-10-9-21-17-22(29)13-15-27(21,5)26(23)14-16-28(24,25)6/h7-10,18-20,22,24-26,29H,11-17H2,1-6H3/b8-7+/t19-,20+,22-,24+,25-,26-,27-,28+/m0/s1

(3S,9S,10R,13R,14R,17R)-17-[(E,2R,5R)-5,6-dimethylhept-3-en-2-yl]-10,13-dimethyl-2,3,4,9,11,12,14,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-3-ol

Provitamin D2; Provitamin D; Ergosterol

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 : ~3.57 mg/mL (~9.00 mM)
Ethanol : ~2.6 mg/mL (~6.55 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.)