Description:FIELD OF INVENTION
The present invention relates to the field of pharmaceutical formulations, specifically to nanoemulsion-based delivery systems for enhanced topical delivery of bioactive compounds, such as kaempferol. It focuses on improving stability, permeation, and therapeutic efficacy for skin-related applications.
BACKGROUND OF THE INVENTION
The growing demand for effective topical treatments for skin-related conditions has highlighted the need for delivery systems that can enhance the stability and bioavailability of active compounds. Conventional topical formulations often face challenges such as poor permeation through the stratum corneum, limited stability of bioactives, and inadequate therapeutic efficacy. These limitations reduce the effectiveness of treatments, especially when using natural compounds like kaempferol, which is known for its potent antioxidant, anti-inflammatory, and antimicrobial properties. However, kaempferol’s low solubility and instability under environmental conditions further hinder its clinical application.
Existing topical delivery systems, including creams and gels, often fail to achieve the desired penetration depth or sustained release of active compounds. This results in suboptimal therapeutic outcomes, requiring frequent reapplication and higher doses, which may lead to undesirable side effects. Furthermore, the use of synthetic chemicals in many formulations raises concerns about potential toxicity and environmental harm. There is a pressing need for a novel formulation approach that addresses these challenges while maintaining safety, efficacy, and user compliance.
Nanoemulsion-based delivery systems have emerged as a promising solution to overcome these limitations. They offer unique advantages such as improved solubility, enhanced skin permeation, and controlled release of active ingredients. By leveraging nanoemulsion technology, the present invention focuses on formulating a stable and efficient topical delivery system for kaempferol, ensuring enhanced therapeutic performance and addressing the limitations of existing formulations. This approach not only improves the bioavailability of kaempferol but also provides a safer and more sustainable alternative for treating various skin conditions.
The following Prior art being Reported was:
IN202211027483: A topical nano emulsion-based nanogel composition for the healing of wounds is disclosed. sprinkler irrigation system is disclosed. The composition comprises a nanogel, wherein the nanogel comprises a nano emulsion, a gelling agent, and a buffer. the nano emulsion comprises lidocaine; oil; emulsifier; and water. The topical nano emulsion-based nanogel composition is suitable for wound healing.
OBJECTS OF THE INVENTION
Some of the objects of the present disclosure, which at least one embodiment herein satisfies, are as follows.
It is an object of the present disclosure to ameliorate one or more problems of the prior art or to at least provide a useful alternative
An object of the present disclosure is to topical pharmaceutical formulations, specifically nanoemulsion-based delivery systems for enhanced therapeutic efficacy of bioactive compound, kaempferol.
Another object of the present disclosure is to enhance the drug stability and encapsulation efficiency, ensuring effective delivery of kaempferol.
Still another object of the present disclosure is to optimized particle size and charge to enhance the stability of the nanoemulsion.
Another object of the present disclosure is to develop a formulation having better skin absorption and retention for targeted therapeutic benefits.
Still another object of the present disclosure is to controlled and sustained drug release for longer-lasting effects.
Still another object of the present disclosure is to develop a smooth texture and ideal thickness make it easy to apply and comfortable to use.
Yet another object of the present disclosure is the formulation process is scalable and economical, making it suitable for commercial manufacturing.
Yet another object of the present disclosure is the developed formulation ensures faster drug penetration through the skin, providing quicker therapeutic relief.

Other objects and advantages of the present disclosure will be more apparent from the following description, which is not intended to limit the scope of the present disclosure.

SUMMARY OF THE INVENTION
The following presents a simplified summary of the invention in order to provide a basic understanding of some aspects of the invention. This summary is not an extensive overview of the present invention. It is not intended to identify the key/critical elements of the invention or to delineate the scope of the invention. Its sole purpose is to present some concept of the invention in a simplified form as a prelude to a more detailed description of the invention presented later.

The present invention is generally focuses on the development of kaempferol-loaded nanoemulsions-gels for topical applications, optimized through various formulation techniques like Box-Behnken experimental design.
An embodiment of the present invention is nanoemulsions were formulated using surfactants (Tween 80), oils (Sefsol), and water phases, with different concentration levels to optimize particle size, stability, and drug release.
Another embodiment of the invention is the kaempferol nanoemulsions demonstrated high drug encapsulation efficiency, ensuring effective incorporation of the active ingredient within the emulsion system for improved delivery.
Yet another embodiment of the invention is the formulations were characterized for key physicochemical properties, including particle size, zeta potential, viscosity, and drug content.
Yet another embodiment of the invention is in-vitro drug release studies were conducted using a dialysis tube method, showing controlled drug release over time and confirming the efficacy of the formulation.
Yet another embodiment of the invention is ex vivo permeation studies indicated the effective transdermal delivery of kaempferol from the nanoemulsion formulations, demonstrating potential for topical application.
Yet another embodiment of the invention is Kaempferol-loaded nanoemulsion gel formulations were created by incorporating the nanoemulsions into Carbopol-based gels, optimizing texture, spreadability, and stability.
Yet another embodiment of the invention also includes skin irritation and erythema studies, confirming the biocompatibility and non-irritant nature of the kaempferol-based formulations for safe use in topical applications.
Yet another embodiment of the invention topical nano emulsion-based nanogel composition is suitable for the treatment of skin-related disorders, including but not limited to inflammation, oxidative stress, and wound healing.
BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1: Transmission Electron Microscopy (TEM) image of the optimized nanoemulsion, illustrating the morphology and size distribution of the nanoparticles.
Figure 2: Cumulative percentage of drug release for all prepared formulations, showcasing the drug release profile over time.
Figure 3: Ex vivo skin permeation results at various time intervals, demonstrating the effectiveness of the formulations in penetrating the skin.
Figure 4: Skin irritation study conducted on mice, presenting the effects of the formulations on skin irritation.
Figure 5: Image showing the skin erythema observed during the skin irritation study, illustrating the level of redness and irritation.
Figure 6: Scoring of erythema severity, categorizing the extent of redness observed during the skin irritation study.
Figure 7: Ear erythema results, highlighting the severity of erythema on the ears of mice after application of the formulation.
Figure 8: Histopathological studies of the prepared optimized nanogel, showing tissue analysis and assessing the biocompatibility and safety of the formulation.

DETAILED DESCRIPTION OF THE INVENTION
The following description is of exemplary embodiments only and is not intended to limit the scope, applicability or configuration of the invention in any way. Rather, the following description provides a convenient illustration for implementing exemplary embodiments of the invention. Various changes to the described embodiments may be made in the function and arrangement of the elements described without departing from the scope of the invention.
The present invention relates to the development of kaempferol-based nanoemulsion-gel formulations designed for enhanced topical application and therapeutic efficacy. Kaempferol, a bioactive flavonoid known for its potent antioxidant and anti-inflammatory properties, serves as the core active ingredient in these formulations. The nanoemulsion was developed using a Box-Behnken design to optimize the concentration of key components such as surfactant (Tween 80), oil phase (Sefsol), and aqueous phase (water). The optimized nanoemulsion demonstrated desirable properties for drug delivery, including fine globule size and enhanced stability. Incorporating this nanoemulsion into a gel base with Carbopol as a gelling agent resulted in the formulation of a nanogel that combines ease of application with sustained drug release.

EXAMPLE 1: Formulation optimization by using Box-Behnken Experimental Design
A Box-Behnken design (BBD) was used to develop the models using Design Expert 13 software. In this study, three independent and three dependent variables were obtained from 17 experiments involving three central points as shown in the Table 1. and their actual and coded values were described in the table as its aids in determining the experimental error and the design precision. The low and high Values for the independent variables were as follows: surfactant (Tween 80), i.e., 15-35% A; oil phase (Sefsol), i.e., 5 – 25 %, B; water (aqueous phase) C i.e., 40-70 %. The mean globule size (Y1), polydispersity index (Y2) and pH (Y3). The table displays the variables chosen according to the Design-Expert software, Lack of fit, ANOVA, and multiple correlation coefficient (r2) tests were used to validate the 3D models.

Table 1 Factor level (independent variable) and response (dependent variable) data for Box-Behnken Design (BBD).
Factor level used Actual (coded)

Independent Variable Low (-1) Medium (0) High (+)
A= Surfactant (%)
B= Oil (%)
C=Water (%) 15
5
40 25
15
55 35
25
70

Another embodiment of the present invention is composition of prepared Formulations are as follows:
Ingredients Pure KMF F1 F2 F3 F4 (Claimed formulation)
Kaempferol % w/v 0.1 0.1 0.1 0.1 0.1
Sefsol % v/v - 25 5 5 15
Tween 80 % v/v - 12.5 12.5 17.5 12.5
PEG 400 %v/v - 12.5 12.5 17.5 12.5
Ethanol % v/v - 10 10 10 10
Carbopol % w/v - - - - 1
Water 100 100 100 100 100

EXAMPLE 2: Characterization of Kaempferol Nanoemulsion Formulations
Kaempferol nanoemulsion formulations were stored at 8°C temperature and later it was characterized for thermodynamic stability studies, particle size, encapsulation efficiency, drug content, and percentage drug release.
Particle size, polydispersity index & zeta potential
Particle size of nanoemulsion and PDI was evaluated using Zeta sizer at 25°C. The prepared nanoemulsion was diluted with distilled water about 1 gm of sample in 1000 ml of distilled water before measurement. The zeta potential of the formulation was carried out by diluting the formulation with 1 mM NaCl in the ratio of 1:1000 with the help of a zeta sizer. All the analysis was carried out in triplicate by using a zetasizer at 25° C.
Results shows that the mean particle size of the nanoemulsion formulation was found to be 332.5 nm and PDI was found to be 0.311. The zeta potential of nanoemulsion formulation F4 was found to be -16.7mV which shows the particles prevents aggregation &therefore stability of nano formulation would be achieved.
Drug Content determination of Kaempferol Nanoemulsion
The drug content determination of nanoemulsion containing drug was incorporating about 5 mg nanoemulsion in 10 ml of methanol, followed by vortexing for 30 minutes for 2 hours. By using 0.45 millipore of filter papers the obtained suspension was filtered, if any insoluble particles, were present again, diluted with methanol 10 ml and measured at 368 nm. Same procedure repeated for blank nanoemulsion.
Results shows that drug content of all prepared formulations were in the range of the 89.69% and 89.75%. This high and uniform drug content across formulations indicates the precision and reproducibility of the preparation method. The formulation F1 achieved the highest percentage drug content at 89.75%.
(% EE) Entrapment Efficiency determination
For determination of entrapment efficiency, kaempferol nanoemulsion of 0.5 ml was diluted with methanol (0.5 ml), and followed for 15 minutes centrifugation at 3500 rpm, obtained supernatant was collected, diluted, and followed for UV-visible spectrophotometer analysis at 368 nm. The percentage entrapment efficiency was calculated by using the equation.
% EE=(Amount of drug added-Amount of free drug in supernatent)/(Total amount of drug added )×100
Results shows that encapsulation efficiency of the all the prepared formulations were in the range of 74.578% to 77.44%, indicating a consistent and effective incorporation of kaempferol within the nanoemulsion systems. Among the formulations, F4 demonstrated the highest encapsulation efficiency at 77.44%, suggesting superior entrapment.
Transmission Electron Microscopy (TEM)
The Morphological analysis of the optimized nanoemulsion was studied by using transmission electron microscopy. The optimized nanoemulsions sample was sonicated on ultra sonicator for 8-10 minutes. A drop of the sample was poured on a 300/200 mesh carbon-coated copper grid with film side up in a pair of self-clamping forceps, followed by air drying for 10 minutes. After, drying sample was negatively stained with 1% phosphotungstic acid solution. Incubate it for 30 seconds, excess amount of liquid was wiped out with the help of filter paper. At last dry sample was subject to TEM examination.
Fig 1 shows the Transmission electron microscopy of optimized nanoemulsion.
Viscosity of Kaempferol Nanoemulsion
By using the Brookfield viscometer, the viscosity of all prepared samples was determined. The instrument spindle was completely immersed into the sample at 25° C. All reading was taken in triplicate.
Results shows that the viscosity of the all prepared formulations were in the range from 128.65 cps (F4) to 172.63 cps (F1). Results as shown in table 2 shows that the formulation F4 had the least viscous consistency, which could influence the spreadability and ease of application, while F1 exhibited the highest viscosity.
Formulation % Encapsulation Efficiency %Drug Content Viscosity (cps)
F1 74.578 89.75 172.63
F2 74.615 89.71 166.21
F3 75.1 89.69 142.62
F4 77.44 89.73 128.65

EXAMPLE 3: In vitro studies
The dialysis tube (MWCO 12KDa) was utilized for assessing the drug release pattern of pure drug solution and nanoemulsion. The desired length of the dialysis tube was pre-soaked in the phosphate buffer solution of pH 6.8 overnight at room temperature. The drug sample of about 5mg was taken in the dialysis bag, and both ends of the bag were tied. The sample inside the dialysis tube was immersed in a dissolution medium of 100 ml which was maintained at 37° C with constant stirring at 50 rpm for uniform distribution of the released drug in the medium. The samples were withdrawn 1 ml at the definite time interval from the beaker by adding the same amount of fresh medium to maintain sink condition. The sample was collected for 12 hours and then the sample was collected for UV-visible Spectrophotometer analysis at 368 nm. The percentage release was calculated and the graph was plotted against cumulative percentage release and time. The kinetic study was described by using zero-order, first order and Higuchi model.
Results shows that the in-vitro release of all prepared nanoemulsion formulation was found to be 62.29%, 72.29%, 68.96%, 79.88% respectively. Nanoemulsion formulation F4 elicits the higher rate of % drug release and the results was found to be 79.88% for 12 hours. Fig 2 shows the Cummultaive percentage of drug release of all prepared formulations.
Table 3: Kinetic Model of the Prepared Optimized Formulation F4.
Kinetic Models Slope (n) Rate Constant Regression coefficient
Zero Order Plot 6.963 -2.8736 0.9894
First Order Plot 0.1444 0.5246 0.8222
Higuchi Plot 24.383 -16.962 0.8877
Korsmeyer & Peppas Plot 1.3199 0.5264 0.9114

EXAMPLE 4: Ex vivo animal Study
Ex-vivo permeation studies were carried out by using the shaved skin of the mice procured from CSIR-CDRI, Lucknow.
The hair of mice was removed from the dorsal region with the help of an electrical shaver without damaging the skin. The animal used for experiment studies was sacrificed by the cervical dislocation method. The shaven part of the skin was separated from the animal by using surgical blade No. 23. The skin was washed with normal saline and the dermis part of the skin was wiped out 3-4 times. the prepared skin was used for the permeation studies.
Skin permeation studies
The Franz diffusion cells were used for ex vivo permeation study with a diffusion area of 2 cm2. The skin of mice from the dorsal side excised were clamped between the donor and the receptor compartment of Franz diffusion cells with the stratum corneum facing the donor compartments. After that 1 g of nanoemulsion containing 0.1% w/w of kaempferol nanogel was applied to the donor components. A phosphate buffer solution of pH 6.8 was filled into the receptor compartment at the maintained temperature of 37° C with stirring at 100 rpm. The samples were withdrawn at predetermined time intervals of 30 min, 1 ml of solution was withdrawn and the same amount of fresh solution was added into the receptor compartment. The same procedure was repeated for up to 24 hours. Samples collected were filled and subjected to a UV-visible spectrophotometer at 368nm.
Drug retention studies
After skin permeation studies, the remaining amount of drug adhering on to the skin was removed with the help of a spatula. The obtained skin was washed 3-4 times with normal saline solution and with the help of tissue paper for drying and for the removal of the drug on the surface of the skin. The modified microwave techniques were used to determine the amount of drug accumulated in the skin layers by separating the epidermis and the dermis layers. After air drying of skin normal room temperature skin was dried in a microwave for 6-12s, followed by separation of the epidermis layer from the dermis region. In 10mL of ethanol obtained skin was soaked separately for the extraction of the drugs and later followed by homogenization. After homogenization, the dispersion obtained was centrifuged for 5 min at 5000 rpm, and the supernatant collected was used to determine the drug content by UV- visible spectrophotometer.
Ex- Vivo Permeation study shows that the cumulative percentage of drug release of the optimized formulation was 85.72%. Fig 3 shows the results of ex-vivo skin permeation at various time intervals.
EXAMPLE 5: Composition and Preparation of Kaempferol Nanoemulsion gel
Ingredients Pure KMF Placebo For
G1
G2 G3
Kaempferol % w/v 0.1 - 0.1 0.1
Sefsol % v/v - 15 15 15
Tween 80 % v/v - 12.5 12.5 12.5
PEG 400 %v/v - 12.5 12.5 12.5
Ethanol % v/v - 10 10 10
Carbopol % w/v - 1 0.5 1
Water 100 100 100 100
The nanoemulsion of kaempferol gel was prepared by adding 0.5 to 1 gram of Carbopol 940 in 100mL of double distilled water and with continuous stirring using a magnetic stirrer for 30 minutes. 0.5 mL of triethanolamine was added, followed by the addition of 0.1 mL of DMSO. After complete formulation of the gel. Nanoemulsion containing kaempferol formulations (01 w/w) were incorporated in the gel with continuous stirring. The prepared gel was kept in a well-closed plastic container for further investigation.
EXAMPLE 6: Characterization of Nanogel
Physicochemical properties of gel
Developed nanogel formulations were subjected to physicochemical evaluation such as physical appearance, color, homogeneity, and, spreadability. Results as shown in table 4 shows that the all the formulations (G1, G2, and G3) exhibited a clear appearance and homogeneous consistency, reflecting uniformity in their physical characteristics.
Drug content (%)
For determination of drug content, nanoemulsion gel 1 gram was dispersed into 10mL of methanol, and subjected to centrifugation for 15 minutes at 3500 rpm, supernatant was collected and diluted if necessary and followed for a UV-visible spectrophotometer at 368 nm.
Results shows that the drug content across all formulations was consistently high, ranging from 92.6 ± 0.0008% (G3) to 93.0 ± 0.0047% (G2), as shown in table 4.
Viscosity determination of gel
The prepared nanogel formulation was subjected to the determination of viscosity by using a Brookfield Viscometer at 20 rpm and measured at 25°C. Results shows that viscosity of all prepared formulations were in the range of 5210 ± 1.2 cps (G1) to 6805 ± 0.86 cps (G3). G3 exhibited the highest viscosity, indicating a thicker consistency, while G1 had the lowest, suggesting a lighter texture.
pH determination of gel
The gels pH was determined and recorded by using a digital pH meter at room temperature. The gel of about 2 gm was dispersed into 20 ml of double distilled water if any impurities were present filtered using filter paper and the pH was recorded. Results were shown in table 4. The pH values of the formulations ranged between 6.0 ± 0.02 and 6.1 ± 0.00, indicating their suitability for topical application on the skin.
Spreadability
A definite amount of gel was added onto the glass slide with the dimension of about 5 ×5 cm and another glass slide with the same dimensions was added or fallen from 3 cm. The spreadability of the gel was measured by using the equation:
S=M×L/T
where S is spreadability in g.cm/s, M is mass of gel in gm, L is length in cm and, T is time in sec.
The spreadability values varied among the formulations, with G2 displaying the highest spreadability at 6.59 ± 0.43 g.cm/s, followed by G3 (5.92 ± 0.60 g.cm/s) and G1 (5.2 ± 0.50 g.cm/s).
EXAMPLE 7: In -vitro release of kaempferol nanogel
For the in-vitro release of nanogel formulations and pure drug suspension, the Franz diffusion cell was utilized. A dialysis membrane was employed for the diffusion process. The dialysis membrane was secured on the Franz diffusion cell, and about 200 mg of the nanogel formulation, and pure drug suspension were added to it. The phosphate buffer of pH 6.8 was added into the receptor compartment and temperature was maintained at 37 ±0.5°C. At definite time intervals, of 0, 0.5,1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, and 24 hours and fresh buffer solution was replaced to maintain the sink conditions. For determination of drug concentrations in the withdrawn sample was analyzed at 368nm by using a UV-visible spectrophotometer.
The in-vitro drug release percentages showed slight variation among the formulations, with G1 releasing 71.07 ± 0.0784%, G2 releasing 79.96 ± 0.963%, and G3 demonstrating the highest release at 80.88 ± 0.77%.
Based on above all evaluation parameter, it was found that the batch G2 containing Carbopol 940 of about (1 % w/v) has the maximum drug content of 93%, Spreadability (g.cm/s) 6.59±0.43 and it also has the optimum viscosity, when compared with the other concentration of Carbopol 940. Thus the batch B2 was optimized and is used further for studies.
EXAMPLE 8: Skin irritation
For measuring the skin irritation of nanogel, topical application tests were performed on mice. The procedure involved applying approximately 0.5 g of the nanogel to the clean-shaved backs of two mice. After application, with the help of an adhesive bandage, and area was covered with gauze. After 24 hours, the applied nanogel of kaempferol was removed and the mice's skin was examined for edema and erythema. The evaluation procedure was subjected to 48 hours. Based on observation of skin reactions, the reaction was categorized as non-irritant, irritant, and, highly irritant
The skin irritation score (total) was calculated by using the formula:
Average irritation scores=(Erythema reaction score+edema reaction score)/(Time ( hours))
Figure 4 illustrates the skin irritation study conducted on mice. Images of the skin were recorded before applying the nanogel and after 24 and 48 hours. The results confirmed that the nanogel caused no irritation.
In vivo skin erythema activity
For skin erythema studies, female mice of a four-week-old with a weight of 20± 2 g were purchased from CSIR- CDRI, Lucknow and randomly assigned into six groups. In a pathogen-free environment, all female mice were kept according to as standard conditions of laboratories with a temperature of 22 ± 2°C and provided 55 ±5% of relative humidity with ad libitum access to food and water and 12 hours of light and dark cycle. All animals were divided into six groups randomly containing four mice in each group. Group I was considered the normal group (placebo) with no treatment, the disease control (positive) was group II, the standard is group III, group IV was treatment I (o.1 w/v KMF NG), group V treatment II (0.2 % w/v KMF NG) and group VI was treatment III (0.4 % w/v KMF NG). The skin was removed or shaved from the dorsal region of the mice with the help of an electric shaver without damaging or affecting the skin. To promote the erythema and skin lesions in the mice skin with slight modification, about 100 µl of 2% DNCB which was dissolved in acetone applied later on applied on the back side of the skin, about 10 µl of DNCB was applied to the back side of both ears at day -4 [33]. Five days after dorsal hair removal, 0.2% DNCB of concentration 100 µl was applied to the back side of the skin, and 10 µl was applied three times a week for 0- 20 days on the back side of the ears. The group III was given a betamethasone gel of 0.1% was applied on the skin seven times a week on both dorsal skin as well as ear for 3 weeks (0-20 days). The mice in Groups IV, V, and VI were treated with kaempferol nanogel with the concentration (0.1%, 0.2% & 0.4%) applied on the dorsal skin region & the back side of the ears from the eleventh day twice a day. On the last day of experiments, animals Ketamine HCl (50mg/kg, i.p.) were used for anesthesia followed by cervical dislocation.
Skin lesions and ear erythema
The mice were anesthetized and later sacrificed by administering ketamine HCl (50 mg/kg, i.p), the skin from the mice was collected from each group and washed with normal saline. With the help of the digital camera, skin lesion images were captured on the last day of the study. Erythema was assessed weekly or in days using a modified scoring system with values of 0, 1, 2,3 and, 4 which are later recorded based on grades as none, slight, moderate, marked, and very marked based on symptoms which are as follows erythema, edema, excoriation/erosion and dryness or scaling. The thickness of mice ears and erythema was recorded after measuring weekly.
Figure 5 shows skin erythema, while Figure 6 depicts the scoring of erythema severity. In Figure 7, ear erythema.
EXAMPLE 9: Histopathological study
On the last day animal study experiment, animals were sacrificed by administering ketamine HCl in the concentration of 50 mg/kg intraperitoneally. After euthanization, the skin of animals was collected from each group. The collected animal skins were then later fixed with 10% formalin solution and it was processed for histopathological examination. The skin tissues were cut into different layers by using a microtome instrument, hematoxylin-eosin was used for staining, and later it was subjected to microscopic examination
Histopathological images show normal skin conditions in Groups I and III with no erythema, while Group II exhibited excessive hypersensitivity, indicating disease induction. The KMF nanogel-treated group displayed improved skin conditions with mild keratosis due to better penetration and slow KMF release. Groups IV to VI showed reduced hypersensitivity and improved skin health, accompanied by hair growth, as shown in figure 8.
EXAMPLE 10: Efficacy Data
Ingredients Pure KMF F1 F2 F3 F4 (Claimed formulation)
Kaempferol % w/v 0.1 0.1 0.1 0.1 0.1
Sefsol % v/v - 25 5 5 15
Tween 80 % v/v - 12.5 12.5 17.5 12.5
PEG 400 %v/v - 12.5 12.5 17.5 12.5
Ethanol % v/v - 10 10 10 10
Carbopol % w/v - - - - 1
Water 100 100 100 100 100
Evaluation of various test formulations
Nanoemulsion formulation No Yes Yes Yes Yes
Emulsion stability - Stable Stable Stable Stable
Entrapment Efficiency (%) - 78.40% 86.71% 84% 94.83%
% of drug release after 12 hours 38% 62% 68% 67% 79%
Evaluation Characteristics of Nanoemulgel (Nanoemulsion Gel) (F4)
Ingredients Pure KMF Placebo For G1 G2 G3
Kaempferol % w/v 0.1 - 0.1 0.1
Sefsol % v/v - 15 15 15
Tween 80 % v/v - 12.5 12.5 12.5
PEG 400 %v/v - 12.5 12.5 12.5
Ethanol % v/v - 10 10 10
Carbopol % w/v - 1 0.5 1
Water 100 100 100 100
Appearance Yellow colour suspension Clear clear Clear
Homogeneous Homogeneous Homogeneous Homogeneous Homogeneous
Viscosity (cps) - 4913±0.64 5210 ±1.2 5732±0.07
Drug content (%) - - 92.8±0.00 93.0± 0.0047
Spreadability (g.cm/s) - 5.0±0.84 5.2±0.50 6.59±0.43
p H 6.0±0.06 6.0±0.02 6.1±0.00
Skin Irritation study - No irritation - No irritation

While considerable emphasis has been placed herein on the specific features of the preferred embodiment, it will be appreciated that many additional features can be added and that many changes can be made in the preferred embodiment without departing from the principles of the disclosure. These and other changes in the preferred embodiment of the disclosure will be apparent to those skilled in the art from the disclosure herein, whereby it is to be distinctly understood that the foregoing descriptive matter is to be interpreted merely as illustrative of the disclosure and not as a limitation.
, Claims:We Claim,
1. A pharmaceutical kaempferol nanoemulsion gel composition comprising of:
a) kaempferol in an amount of 0.1% w/v;
b) sefsol in an amount of 15% v/v;
c) tween 80 in an amount of 12.5% v/v;
d) poly ethylene glycol 400 in an amount of 12.5% v/v;
e) ethanol in an amount of 10% v/v;
f) carbopol in an amount ranging from 0.5% to 1% w/v; and
g) water as the balance to 100%.
Wherein the active ingredient kaempferol is loaded in the form of nanoemulsion;
wherein the nanoemulsion has a particle size of less than 200 nm, and the composition is formulated for topical application with enhanced stability, bioavailability, and therapeutic efficacy.
2. The method for preparing a kaempferol nanoemulsion gel, as claimed in claim 1, wherein the step comprising of:
a) dispersing 0.5% to 1% w/v of Carbopol 940 in double-distilled water with continuous stirring for 30 minutes using a magnetic stirrer;
b) neutralizing the dispersion by adding 0.5% v/v of triethanolamine to form a gel base;
c) incorporating 0.1% v/v of dimethyl sulfoxide (DMSO) into the gel base under stirring;
d) preparing a nanoemulsion comprising 0.1% w/v kaempferol, 15% v/v sefsol, 12.5% v/v tween 80, 12.5% v/v poly ethylene glycol 400, 10% v/v ethanol, and water as the balance; and
e) blending the nanoemulsion into the gel base under continuous stirring to form a homogeneous nanoemulsion gel, which is stored in a well-closed container for further use.
3. The pharmaceutical kaempferol nanoemulsion gel composition, as claimed in claim 1, wherein the optimized composition of kaempferol nanoemulsion gel, comprising of:
a) kaempferol in an amount of 0.1% w/v;
b) sefsol in an amount of 15% v/v;
c) tween 80 in an amount of 12.5% v/v;
d) poly ethylene glycon 400 in an amount of 12.5% v/v;
e) ethanol in an amount of 10% v/v;
f) carbopol in an amount of 1% w/v; and
g) water as the balance to 100%.
4. The pharmaceutical kaempferol nanoemulsion gel composition as claimed in claim 1, wherein the gel provides sustained release of kaempferol over a period of 8 to 12 hours.
5. The pharmaceutical kaempferol nanoemulsion gel composition as claimed in claim 1, wherein the pH of the gel is in the range of 5.5 to 7.5.

Dated this 16 January 2025

Dr. Amrish Chandra
Agent of the applicant
IN/PA No: 2959