Lanosterol 14-alpha demethylase (CYP51); Ergosterol biosynthesis; p38 MAPK; COX-2; PGE2; Microtubules; Candida albicans; Dermatophytes

Sertaconazole (0.03-40 µg/mL; 24 h) inhibits 150 yeast strains, including six species of Candida, whose arithmetic mean minimum inhibitory concentration (MIC) is 0.77 µg/mL [1]. In a time-dependent manner, p38 MAP kinase is activated by sertaconazole (1 µg/mL; 5, 10, 30, 60 min)[2]. Dependent on p38 activation, sertaconazole (1, 2 µg/mL; 6, 8, or 24 h) causes a twofold release of PGE2 via COX-2 in keratinocytes[2]. By depolymerizing interphase and spindle microtubules, cetaconazole (10, 20, 30, 40 µM; 24 h) promotes significant mitotic arrest, which results in chromosomal aggregation problems and an anti-proliferation effect[3]. In HeLa cells, sertaconazole (20, 40 µM; 24 h) causes apoptosis via the p53 pathway[3]. HeLa cell migration is inhibited by sertaconazole (20, 30 µM; 24, 48, and 72 h) in a concentration-dependent manner[3]. In A549 and H460 cells, sertaconazole (15, 30 µM; 24 h) initiates autophagy[4].

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Sertaconazole inhibits ergosterol synthesis in Candida albicans with an IC50 of 115 nmol/L. It is fungistatic and fungicidal in vitro with MICs ranging from 0.35 to 5.04 μg/mL and 0.5 to 16 μg/mL, respectively. The compound exhibits anti-inflammatory activity by activating the p38-COX-2-PGE2 pathway. It also acts as a microtubule inhibitor, showing antiproliferative effects. It inhibits mouse IDO2 in HEK293T cells with ≥55% inhibition at 20 μM.

Sertaconazole (1% (w/v); applied once to the left ear) reduces TPA-induced otitis media in CD-1 mice[2].

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In vivo, sertaconazole is used as a topical antifungal for cutaneous mycoses, including dermatophytosis, candidiasis, and pityriasis versicolor. It achieves high local concentrations in the skin with minimal systemic absorption. Clinical studies demonstrate efficacy comparable to other topical antifungals with a favorable safety profile. The anti-inflammatory activity contributes to rapid symptom relief in inflammatory fungal infections.

For ergosterol synthesis inhibition assays: culture Candida albicans ATCC E-10.231 at 6 × 10⁵ cells/mL in aerobic conditions at 37°C. Add sertaconazole at varying concentrations (1–1000 nmol/L) and measure ergosterol content by GC-MS or HPLC. Calculate IC50. For enzyme binding: incubate recombinant CYP51 with sertaconazole and a fluorogenic substrate, measure fluorescence.

Cell Viability Assay[1]
Cell Types: C. albicans, C. guilliermondii, C. krusei, C. parapsilosi, C. tropicalis, C. glabrata
Tested Concentrations: 0.03-40 µg/m
Incubation Duration: 24 h
Experimental Results: Againsted 150 strains of yeasts (six Candida species) which included C albicans, C. guilliermondii, C. krusei, C. parapsilosi, C. tropicalis, C. glabrata species with arithmetic mean MIC values of 1.02, 0.51, 0.38, 0.31, 1.67 and 0.78 µg/mL, respectively.

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Western Blot Analysis[2]
Cell Types: HaCaT cells
Tested Concentrations: 1 µg/mL
Incubation Duration: 5, 10, 30, 60 min
Experimental Results: demonstrated activity of activating p38 MAP kinase and Hsp27 in a time-dependent manner.

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Western Blot Analysis[2]
Cell Types: HaCaT cells
Tested Concentrations: 1, 2 µg/mL
Incubation Duration: 6 or 8 h
Experimental Results: Induced 50% expression of COX-2 and resulted in a twofold increased in PGE2 release.

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Western Blot Analysis[2]
Cell Types: siRNA-transfected HaCaT cells (without p38 MAP kinase expression)
Tested Concentrations: 1 µg/mL
Incubation Duration: 24 h
Experimental Results: Mediated induction o

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For antifungal susceptibility testing: use broth microdilution per CLSI M27-A3 guidelines with RPMI 1640 medium. Prepare Candida albicans inoculum (0.5–2.5 × 10³ CFU/mL), add sertaconazole in 2-fold serial dilutions (0.03–16 μg/mL), and incubate at 35°C for 24–48 hours. Determine MIC as the lowest concentration inhibiting visible growth. For anti-inflammatory assays: measure PGE2 production by ELISA in LPS-stimulated macrophages.

Animal/Disease Models: CD-1 mice (TPA-induced ear edema model)[2].
Doses: 1% (w/v)
Route of Administration: Apply to the left ear, once.
Experimental Results: demonstrated a significant reduction of inflammation in mice by mediating PGE2 release.

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For in vivo antifungal studies: use guinea pig or mouse models of cutaneous candidiasis or dermatophytosis. Apply sertaconazole topically (1–2% cream) once or twice daily for 7–14 days. Assess lesion severity, fungal burden by culture, and histological improvement. For anti-inflammatory studies: use carrageenan-induced paw edema in rats; apply sertaconazole topically and measure paw volume. For pharmacokinetic studies: measure skin and plasma concentrations by HPLC.

Absorption, distribution, and excretion Bioavailability is negligible.

Toxicity Summary
Sertaconazole interacts with 14α-demethylase, a cytochrome P-450 enzyme essential for the conversion of lanosterol to ergosterol. Since ergosterol is a crucial component of the fungal cell membrane, inhibition of its synthesis leads to increased cell permeability, resulting in leakage of cell contents. Sertaconazole may also inhibit endogenous respiration, interact with membrane phospholipids, inhibit yeast conversion to mycelium, inhibit purine uptake, and impair the biosynthesis of triglycerides and/or phospholipids. Protein Binding Plasma protein binding is 99%.

[1]. In-vitro antifungal activity of sertaconazole, econazole, and bifonazole against Candida spp. J Antimicrob Chemother. 1995 Oct;36(4):713-6.

[2]. Anti-inflammatory activity of sertaconazole nitrate is mediated via activation of a p38-COX-2-PGE2 pathway. J Invest Dermatol. 2008 Feb;128(2):336-44.

[3]. Sertaconazole induced toxicity in HeLa cells through mitotic arrest and inhibition of microtubule assembly. Naunyn Schmiedebergs Arch Pharmacol. 2021 Jun;394(6):1231-1249.

[4]. Sertaconazole provokes proapoptotic autophagy via stabilizing TRADD in nonsmall cell lung cancer cells. MedComm (2020). 2021 Dec 16;2(4):821-837.

1-{2-[(7-chloro-1-benzothiophene-3-yl)methoxy]-2-(2,4-dichlorophenyl)ethyl}imidazolium belongs to the imidazolium class of compounds, containing a 2-[(7-chloro-1-benzothiophene-3-yl)methoxy]-2-(2,4-dichlorophenyl)ethyl group at the 1-position. It is a dichlorophenyl ether, belonging to both the imidazolium and 1-benzothiophene classes. Sertaconazole nitrate is an imidazolium antifungal drug. It is available in topical formulations for treating skin infections such as athlete's foot. Sertaconazole is an azole antifungal drug. Sertaconazole is a synthetic imidazolium derivative containing a benzothiophene ring, possessing antifungal, antibacterial, anti-inflammatory, and antipruritic activities. Besides inhibiting ergosterol synthesis, the benzothiophene ring of sertaconazole can insert into the fungal cell wall, replacing tryptophan. This increases cell wall permeability. Furthermore, sertaconazole can inhibit the release of cytokines. Sertaconazole was only detected in individuals who had used or taken the drug. Sertaconazole nitrate is an imidazole antifungal drug used in cream form to treat skin infections such as tinea pedis. [Wikipedia] Sertaconazole interacts with 14α-demethylase, a cytochrome P-450 enzyme responsible for converting lanosterol to ergosterol. Since ergosterol is an important component of the fungal cell membrane, inhibition of its synthesis leads to increased cell permeability, resulting in leakage of cell contents. Sertaconazole may also inhibit endogenous respiration, interact with membrane phospholipids, inhibit the conversion of yeast to mycelium, inhibit purine uptake, and impair the biosynthesis of triglycerides and/or phospholipids.
See also: Sertaconazole nitrate (in salt form).

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Drug Indications
For topical treatment of interdigital tinea pedis caused by Trichophyton rubrum, Trichophyton mentagrophytes, and Epidermophyton floccosum in immunocompetent patients aged 12 years and older.
FDA Label
Mechanism of Action
Sertaconazole interacts with 14α-demethylase, a cytochrome P-450 enzyme essential for the conversion of lanosterol to ergosterol. Since ergosterol is a crucial component of the fungal cell membrane, inhibition of its synthesis leads to increased cell permeability, resulting in leakage of cell contents. Sertaconazole may also inhibit endogenous respiration, interact with membrane phospholipids, inhibit yeast conversion to mycelium, inhibit purine uptake, and impair the biosynthesis of triglycerides and/or phospholipids.
Pharmacodynamics
Sertaconazole is an imidazole/triazole antifungal drug. Sertaconazole selectively inhibits the C-14α-demethylation of fungal cytochrome P450 sterols by inhibiting cytochrome P450 14α-demethylase. This enzyme converts lanosterol to ergosterol, essential for fungal cell wall synthesis.
The subsequent loss of normal sterols is associated with the accumulation of 14α-methylsterol in fungi, which may be partly responsible for the antibacterial activity of fluconazole. Demethylation in mammalian cells is much less sensitive to the inhibitory effects of fluconazole. Sertaconazole exhibits in vitro activity against Cryptococcus neoformans and Candida spp. In normal and immunocompromised animal models, sertaconazole also showed antibacterial activity against systemic and intracranial fungal infections caused by Cryptococcus neoformans and systemic infections caused by Candida albicans.

C20H15N2OSCL3

437.77

435.997

99592-32-2

Sertaconazole nitrate;99592-39-9

65863

Typically exists as solid at room temperature

1.4±0.1 g/cm3

614.1±55.0 °C at 760 mmHg

325.2±31.5 °C

0.0±1.7 mmHg at 25°C

1.675

7.49

0

3

6

27

488

0

C1=CC2=C(C(=C1)Cl)SC=C2COC(CN3C=CN=C3)C4=C(C=C(C=C4)Cl)Cl

JLGKQTAYUIMGRK-UHFFFAOYSA-N

InChI=1S/C20H15Cl3N2OS/c21-14-4-5-16(18(23)8-14)19(9-25-7-6-24-12-25)26-10-13-11-27-20-15(13)2-1-3-17(20)22/h1-8,11-12,19H,9-10H2

1-[2-[(7-chloro-1-benzothiophen-3-yl)methoxy]-2-(2,4-dichlorophenyl)ethyl]imidazole

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)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

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.)