Fungal infections may be classified as superficial infections affecting the skin, hair, nails or mucous membranes, or systemic, affecting the body as a whole. Subcutaneous infection is largely confined to the subcutaneous tissue and dermis, but may extend to the epidermis as well as deeper, for example, to bone; systemic infections tend to occur more frequently in immunocompromised patients such as those with AIDS. Superficial fungal infections are among the most widespread diseases known to man. They target parts of the body as diverse in form and function as the skin, the nail, the buccal cavity, the eye and the vagina.²

Ketoconazole (KTZ) is a broad-spectrum azole antifungal approved in 1981 by the United States (US) Food and Drug Administration (FDA) for systemic fungal infections. Azole antifungals inhibit the P450-dependent enzyme lanosterol 14-α-demethylase, blocking synthesis of ergosterol, an important structural component of fungal cell membranes, and leading to cell death. During the 1980s, KTZ was the only oral antifungal agent available for the treatment of systemic mycoses³. Due to non-specific inhibition of mammalian P450-dependent enzymes, reports of serious adverse events such as hepatotoxicity, adrenal insufficiency, and drug-drug interaction led to the withdrawal of oral KTZ from Europe and Australia, and label changes in North America to remove the indication for treatment of skin and nail fungal infections in 2013. Currently, oral KTZ is only indicated for the treatment of endemic mycoses when alternative antifungal therapies are not available or tolerated⁴. KTZ has antifungal activities against localized, superficial Malassezia, Candida and dermatophyte infections, leading to the development of topical KTZ with the hopes that the drug is not systemically absorbed thus limiting drug-related adverse events. Although animal studies report mild-to-moderate ocular and dermal irritation with topical KTZ, there are no reports of systemic toxicity in animals. To these authors’ knowledge there is no review discussing adverse effects of topical KTZ in humans⁵.

Fig no – 2

Mechanism of Action:

Ketoconazole (NIZORAL, chemical name: cis-1-acetyl4-[4-[[2-(2,4-dichlorophenyl)-2-(1H-imidazol-1-ylmethyl)- 1,3-dioxolan-4-yl]methoxy] phenyl] piperazine is an azolebased antifungal, which targets CYP450 enzymes vital to cell metabolism and detoxification. Ketoconazole principally inhibits lanosterol 14a-demethylase -- the enzyme that regulates the synthesis of ergosterol⁶. The disruption of ergosterol biosynthesis alters cell membrane structure, thus compromising membrane integrity and permeability, and consequently interfering with cellular growth and reproduction. In humans, ketoconazole can cause AEs by interacting with the CYP450 class of enzymes in non-target cells. Ketoconazole is hepatically metabolized and excreted and hepatotoxicity may result through the direct action of the drug and its primary metabolite, N-deacetyl ketoconazole on hepatic cells. Ketoconazole interacts with the human CYP3A isoforms impacting hepatic drug clearance and alters steroid synthesis⁷. Given that it is a potent inhibitor of CYP3A4, concomitant administration of oral ketoconazole with other drugs metabolized by this system can raise serum levels, consequently increasing the toxicity of these agents. Furthermore, CYP450-dependent enzymes are responsible for the conversion of cholesterol to steroid hormones such as testosterone and cortisol, and thus are involved in regulating testicular and adrenal gland function. Despite ketoconazole’s therapeutic benefit, modulation of the CYP450 pathway has widespread physiological implications that pose significant risk for drugrelated AEs⁸.

Classification of Fungal Infection:

1 Superficial:-

Causative Fungus: Trichophyton rubrum.

Aspergillus Fumigatus.

Ex: Tinea versicolor , White piedra , tinea nigra.

Fig no – 3

2. Cutaneous:

Causative Fungus: Epidermophyton, Trichophyton,

Microsporum.

Ex: Tinea faciei, tinea barbae, tinea capitis, tinea manuum.

Fig no – 4

3. Subcutaneous:

Causative Fungus: Sporothrixschencki, Candida albicans .

Ex: Maduramycosis, chromomycosis

Fig no - 5

MATERIALS AND METHODS:

Table no – 1

Sr. No	Ingredients	Obtained from
01	Ketoconazole	From the central store of Kle College of pharmacy, nipani
02	Eucalyptus oil	From the central store of Kle college of Pharmacy, nipani
03	Poloxamer	From the central store of Kle college of Pharmacy, nipani
04	Propylene Glycol	From the central store of Kle College of pharmacy, nipani
05	Potassium Sorbate	From the central store of Kle College of Pharmacy, nipani
06	Lecithin	From the central store of Kle College of Pharmacy, nipani
07	Isopropyl Myristate	From the central store of Kle college of Pharmacy, nipani
08	Castor Oil	From the central store of Kle college of Pharmacy, nipani
09	Paraffin Oil	From the central store of Kle College of pharmacy, nipani

FORMULATION OF KETOCONAZOLE GEL:

Table no - 2

Sr. No	Ingredients	Components (%)
01	Ketoconazole	2
02	Eucalyptus oil	5
03	Poloxamer	15.5
04	Propylene Glycol	10
05	Potassium Sorbate	0.2
06	Lecithin	5
07	Isopropyl Myristate	Up to 100
08	Castor Oil	Up to 100
09	Paraffin Oil	Up to 100

PROCEDURE:

STEP 1: AQUEOUS PHASE PREPARATION:

The aqueous phase was received by dissolving a weighed quantity of Pluronic® F-127 (15.5%, w/w) in an aqueous-based solution containing propylene glycol and potassium sorbate.

STEP 2: OIL PHASE PREPARATION:

The oily phase was prepared by dissolving lecithin (5%, w/w) in isopropyl myristate, castor oil, or paraffin oil, respectively. After preparation, the water phase and the oil phase were conditioned at 4 ◦C for 24 h in order to completely dissolve the ingredients and stabilize the systems. Pluronic/lecithin organogels (PLOs)

STEP3: MIXING PHASE:

Combine water and oil phases in different weight ratios, 30:70, 50:50, and 70:30, using a mechanical stirrer at a speed of 400 rpm. A stable and homogeneous gel system was obtained at a weight ratio of water phase to oil phase of 70:30. KTZ and TTO were added to the lipid phase before organogel formation. The compositions of the designed formulations are listed in Table.⁰⁹

EVALUATION OF GEL:

Organoleptic Characteristics Each formulation was organoleptically tested for odor, color, texture, phase separation, and greasiness.

pH Determination:

The pH was measured using the glass electrode of an Orion 3-Star pH Meter (Thermo Scientific, Waltham, MA, USA). Each measurement was carried out six times and the average pH was calculated.

Determination of Rheological Properties:

The viscosity was determined using a Brookfield viscometer (RVDV-III Ultra, Brookfield Engineering Laboratories, Middleboro, MA, USA) equipped with a cone/plate type CPA52Z measuring system (plate diameter: 24mm; cone angle: 3^◦) at a temperature of 25^◦C ±1^◦C. The viscosity values of the gel formulations at shear rate 6.00 s−1 were recorded and the rheograms were estimated by plotting the obtained values of shear stress versus shear rate (2.00 –10.00 s−1).

Fig no - 6

Fig no – 7

Fig no - 8

Fig no – 9

Fig no - 10

RESULT AND DISCUSSION:

Preformulation study:

Characterization of the drug (KETOCONAZOLE):

1. UV spectroscopic study (λ_max):

The λ_maxof drug was found to be 205nm. The UV spectrum of the drug solution is shown in Fig. 11

Melting point:

The melting point of pure drug was first determined by capillary method and the melting point was found to be in the range 168-170⁰C which matches with the reported value Ketoconazole.

The melting point of Ketoconazole was found to be around 149.79C.

Development and Validation of UV Spectroscopic method:

Preparation of standard stock solution:

Accurately weighed 100mg of pure Ketoconazole drug was transferred into a 100ml volumetric flask. The small quantity of methanol was added to ensure complete solubilization of drug and finally, volume was made up to the mark with Phosphate buffer 6.8³⁸. The clear solution was obtained with the strength of 1000µg/ml. From this stock solution, 10ml was taken into a 100ml volumetric flask, diluted up to 100ml with Phosphate buffer 6.8 to get working solution of 100μg/ml concentration and filtered through Whatman filter paper¹⁰.

Preparation of sample solutions:

Different aliquots of stock solution 1 to 6ml were transferred to 10ml volumetric flasks and diluted up to mark to get solution of 10-60µg/ml. These dilutions were scanned at 205nm. The correlation coefficient and regression line equation for Ketoconazole were determined by plotting the calibration curve of absorbance vs. concentration.

Standard calibration curve of ketoconazole by UV spectroscopic method:

The standard calibration curve of Ketoconazole was determined by the absorbance of Ketoconazole solution in Phosphate b 6.8 buffer having range of 10-60µg/ml. The concentration v/s absorbance graph was plotted by using values given in table no.18. From this plot Ketoconazole showed linearity with R²of 0.999 and standard equation of line was obtained and used for further analysis.

Table no – 3

Conc (µg/ml)	Absorbance
10	0.98
20	1.35
30	1.77
40	2.15
50	2.57
60	2.96

Fig no – 12

Table No - 4

S. No	Tested solvent	Solubility of KTZ (ug/ ml)
1	Water	9.56 ±0.09
2	Isopropyl myristate	93.99±3.26
3	Caster oil	93.55±8.39
4	Paraffin oil	17.02± 2.84
5	Water+Eucalyptus oil 5%	35.70±4.59
6	Isopropyl myristate+Eucalyptus oil 5%	130.95± 5.13
7	Caster oil+Eucalyptus oil 5%	154.78±7.94
8	Paraffin oil+Eucalyptus oil 5%	80.11±6.5

FORMULATION DEVELOPMENT:

Solubility:

The solubility of KTZ was determined by using the shake-flask method. In brief, an excess amount of KTZ was added to each vial containing 5mL of the selected solvents (water, isopropyl myristate, castor oil, paraffin oil, and the abovementioned solvents with the addition of 5% eucalyptus oil). Mixtures were mechanically shaken for 24h at 25^◦C±0.5^◦C and allowed to stand for 24 h to attain equilibrium. Then, the samples were centrifuged at 4000rpm for 15min, followed by filtration through a CA membrane filter (0.45µm), diluted appropriately with methanol.

Solubility of KTZ in chosen solvents. Values are expressed as means ± SD of six replicates:

Ph:

The pH values were found in the acceptable range and thus would cause no irritation. The pH was found in the range of 6.6 – 7.8.

Viscosity:

The viscosity of the formulated gel was found to be 3883 cP.

ACKNOWLEDGMENT:

For the completion of this dissertation work, many people have contributed by helping us throughout the course. It is a privilege to express our heartfelt thanks to all those who have contributed directly or indirectly to the success of this dissertation work. It is our pleasant duty to acknowledge our deepest sense of gratitude to K.L.E ‘s College of Pharmacy, Nipani for his invaluable and consistent guidance and suggestions which always stimulated to pursue the investigation with great interest.

We wish to express our deep sense of gratitude and sincere thanks to Dr. Pramod C. Gadad,

Principal K.L.E ‘s College of Pharmacy, Nipani for his constructively criticizing advice and support while doing our work

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Received on 27.03.2025 Revised on 28.04.2025

Accepted on 25.05.2025 Published on 30.10.2025

Available online from November 08, 2025

Research J. Topical and Cosmetic Sci. 2025; 16(2):91-96.

DOI: 10.52711/2321-5844.2025.00015

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