A niosomal formulation of bioactive component for topical application and a process of preparation thereof

Abstract

A niosomal formulation of bioactive component for topical application and a process of preparation thereof [0069] The invention discloses a niosomal formulation of bioactive components for topical application. The niosomal formulation comprises non-ionic surfactants, cholesterol, glycerol, water, bioactive component propylene glycol and polysachharides. The process of preparation of niosomal formulation comprises steps of mixing cholesterol and non-ionic surfactants, homogenizing the mixture, mixing the bioactive component, propylene glycol and glycerol, nano milling the mixture, nano milling the homogenized and nano milled mixture to obtain multilamellar niosomes, homogenization to obtain large unilamellar niosomes, homogenization to further obtain small unilamellar niosomes. The process involves optimized temperatures favoring the formation of stable niosomes. The niosomal formulation is topically administered and does not induce any skin irritation. The niosomal formulation is safe, bioavailable and highly stable.

Description

Internal Ref: OR26C021PCT03TITLE OF THE INVENTIONA niosomal formulation of bioactive component for topical application and a process of preparation thereofPriority Claim

[0001] This application claims priority from the provisional application numbered 202441072308 filed with Indian Patent Office, Chennai on 25thMarch 2025 entitled ‘M niosomal formulation of bioactive component for topical application and a process of preparation thereof", the entirety of which is expressly incorporated herein by reference.Preamble to the Description

[0002] The following specification describes the invention and the manner in which it is to be performed:DESCRIPTION OF THE INVENTIONTechnical field of the invention

[0003] The present invention relates to a niosomal formulation comprising bioactive component for topical application and a process of preparation thereof. More specifically, the invention discloses a process of preparing the niosomal formulation that exhibits enhanced stability, improved skin compatibility, and effective delivery of the bioactive component upon topical administration.Background of the invention

[0004] Niosomes are vesicles composed of non-ionic surface-active agent bilayers, which serve as effective drug delivery systems. Niosomes are microscopic in size. Niosomes are formed on the admixture of non-ionic surfactant and cholesterol with subsequent hydration in aqueous media. Niosomes are unilamellar or multilamellar depending on the method used to prepare them. Niosomes aid in achieving targetedInternal Ref: OR26C021PCT03delivery, better drug entrapment efficiency and to develop specialized niosomes with special structures.

[0005] The niosome is made of a surfactant bilayer with its hydrophilic ends exposed on the outside and inside of the vesicle while the hydrophobic chains face each other within the bilayer. The vesicle holds hydrophilic drugs within the space enclosed in the vesicle while the hydrophobic drugs are embedded within the bilayer itself.

[0006] Niosomes are categorized according to their lamellarity. The bilayered vesicles may be Small Unilamellar Vesicles (SUV), Large Unilamellar Vesicles (LUV) or Multi-Lamellar Vesicles (MLV). Different sizes of niosomes are necessary for the optimization of specific delivery requirements, and several different approaches are used to produce these nanocarrier systems.

[0007] Various methods have been developed for synthesizing niosomes from ingredients mentioned in the previous section according to their particle size, lamellarity, and clinical applications. The methods incorporated for the synthesis of niosiomes are bubble method, ether injection method, hand shaking method, heating method, microfluidization method, multiple membrane extrusion method, reverse phase evaporation method, sonication method, thin film hydration method, and transmembrane pH gradient method.

[0008] Niosomes offer many advantages over conventional drug delivery systems in terms of flexibility for drug delivery and the capability to entrap hydrophilic and hydrophobic drugs. Niosomes entrap solutes and API of different solubilities, delivering them via many different routes of administration: oral, pulmonary, ocular, parenteral and topical. The surfaces of the vesicle structure can be functionalised for targeted delivery, and therapeutic performance may be enhanced due to reduced clearance from the biological system. The surfactants used in the synthesis of niosomes are biodegradable, biocompatible and nonimmunogenic, making them ideal for human use. Niosomes are osmotically active and stable and enhance the stability of the payload. Niosomes exhibit enhanced stability and overall circulation half-life.Internal Ref: OR26C021PCT03

[0009] The patent application No. US10251820B2 entitled “Topical composition comprising plant extracts” discloses a topical composition comprising plant extracts embedded in hydrophilic vesicles niosomes, having a size smaller than 500 nm. The niosomes are composed of amphiphilic molecules selected from the group consisting of a linear polyglycerol esterified with a saturated linear fatty acid, a branched polyglycerol esterified with a saturated linear fatty acid, a linear polyglycerol esterified with a mono-unsaturated linear fatty acid, a branched polyglycerol esterified with a mono-unsaturated linear fatty acid. The noisome niosomes comprise a stabilizer consisting of glycerine and a hydrosoluble natural antioxidant. The plant extract is an extract of a plant selected from the group consisting of Norway spruce, red clover, ginkgo biloba, and bilberry. The excipient is selected from a group comprising an emulsifier, a hydrating agent, a solvent, an emollient, a stabilizer, a viscosity modifier, a preservative, a lubricant, a sequestering agent, a chelating agent, a filler, a powder, a fragrance, a perfume, an absorbent, a dye, an opacifier, an antioxidant, a vitamin, a screening substance, a UV filter, an essential oil, a keratin-active substance, and an amino acid. The plant extract is embedded in an aqueous compartment delimited by a double layer of said amphiphilic molecules. The process of preparation of niosomes comprises steps of forming niosomes by hand shaking or ultrasonification of an aqueous mixture which comprises 35% to 50%, based on the weight of said aqueous mixture, of an amphiphilic compound selected from the group consisting of a linear polyglycerol esterified with a saturated linear fatty acid, a branched polyglycerol esterified with a saturated linear fatty acid, a linear polyglycerol esterified with a monounsaturated linear fatty acid, a branched polyglycerol esterified with a monounsaturated linear fatty acid.

[0010] The patent application No. IN202321041384A entitled “Niosomal antifungal topical gel formulation for drug delivery” discloses a niosomal antifungal topical gel of Leonotis nepetaefolia and a process for the preparation of niosomal antifungal topical gel, comprising of preparing niosomes comprising of mixing 2: 1 ratio of cholesterol and Tween 40 and dissolving in 5 mL of chloroform, evaporating lipid solution by rotating the said flask at 135 rpm to form a smooth and dry lipid film and hydrating the film with 5 ml phosphate buffer saline of pHInternal Ref: OR26C021PCT037.4 for 3 hours with gentle shaking to form niosomes and stabilizing it by keeping at temperature 2°C-8°C for 24 hours, preparing niosomal suspension of Leonotis nepetaefolia extract comprising of dissolving 2.5 gm of methanolic extract of Leonotis nepetaefolia in 9.6 ml of propylene glycol to form 2.5% w / w methanolic extract of Leonotis nepetaefolia and adding the extract to the niosomes to form Leonotis nepetaefolia encapsulated niosomal suspension, preparing niosomal gel formulation of Leonotis nepetaefolia extract comprising of dispersing 2gm of carbopol 934 in distilled water and neutralizing it by adding triethanolamine to obtain a clear gel, adding niosomal suspension of Leonotis nepetaefolia into said dispersion by gentle mechanical mixing to obtain niosomal antifungal topical gel of Leonotis nepetaefolia. The niosomal antifungal topical gel has methanolic extract: surfactant: cholesterol is in the ratio of 1 :2: 1. The niosomal gel formulation wherein the particle size 2.82±1.09 pm, entrapment efficiency 84.84±0.36% and drug content 98.28±0.11. The niosomal gel formulation is prepared using the flowers of Leonotis nepetaefolia. The preparation process of methanolic extract of Leonotis nepetaefolia, comprising of collecting, dehydrating, pulverizing, extracting Leonotis nepetaefolia flowers in a hot continuous extraction mode using methanol as the solvent, removing leftover dissolvent after collecting the methanol dissoluble components in the receiver and storing the product.

[0011] Although various formulations and methods for the synthesis of niosomes exist, they bear certain drawbacks. The existing formulations upon administration cause possible skin irritation, exhibit lesser load ability of bioactive ingredients, leakage of bioactive from the niosome leading to product loss and extended formulation time. Additionally due to the mentioned drawbacks physical and chemical stability of the niosomes are hindered. Hence, there is a requirement of a niosomal formulation and a method to develop the niosomal formulation to overcome the existing shortcomings.Summary of the inventionInternal Ref: OR26C021PCT03

[0012] The present invention overcomes the drawbacks of the conventional systems to disclose a niosomal formulation of bioactive components for topical application and a process for preparation thereof.

[0013] The niosomal formulation comprises non-ionic surfactants, cholesterol, glycerol, propylene glycol, water, and at least one bioactive component selected from, but not limited to vitamin C, vitamin B3, and hyaluronic acid.

[0014] According to an embodiment, the present invention also discloses a process for preparing nano-niosomal formulation comprising sequential steps of preparing a lipid phase including cholesterol and non-ionic surfactants, preparing polysaccharide solution by dissolving gum ghatti, xanthan gum, guar gum, pectin, etc in water, preparing an aqueous phase comprising glycerol and water, and preparing an active phase comprising the bioactive component dissolved in a solvent system. The process further comprises subjecting the active phase to nanomilling, followed by mixing with the lipid phase and subsequent nano-milling to form multilamellar niosomes. The multilamellar vesicles are further subjected to ultrasonication to obtain large unilamellar niosomes, and subsequently nano-sized unilamellar niosomes having reduced vesicle size and improved uniformity.

[0015] The present invention provides a vitamin C-loaded nano-niosomal formulation exhibiting high encapsulation efficiency and loading capacity, along with improved physical stability. Analytical evaluation confirms effective incorporation of the bioactive component within the vesicular system with minimal degradation.

[0016] Further, the in vitro studies demonstrate that the nano-niosomal formulation exhibits reduced cytotoxicity compared to the corresponding free bioactive component and maintains high cell viability. Further, permeability studies indicate significantly enhanced permeation across epithelial barriers, with an apparent permeability coefficient substantially higher than that of the free bioactive component, thereby supporting improved transdermal delivery.Internal Ref: OR26C021PCT03

[0017] The niosomal formulation of the present invention provides controlled and sustained release of the bioactive component, enhanced bioavailability, reduced leakage, and improved stability. The formulation is dermatologically acceptable and suitable for topical, pharmaceutical, nutraceutical, and cosmeceutical applications.Brief description of the drawings

[0018] The foregoing and other features of embodiments will become more apparent from the following detailed description of embodiments when read in conjunction with the accompanying drawings. In the drawings, like reference numerals refer to like elements.

[0019] Figure 1 tabulates the niosomal formulation of bioactive components for topical application.

[0020] Figure 2 illustrates the flowchart for the process of preparation of niosomal formulation of bioactive component.

[0021] Figure 3 illustrates the cytotoxicity of niosomal vitamin C formulation of the present invention with vitamin C.

[0022] Figure 4 illustrates the permeability of niosomal vitamin C formulation of the present invention with vitamin C.

[0023] Detailed description of the invention

[0024] In order to more clearly and concisely describe and point out the subject matter of the claimed invention, the following definitions are provided for specific terms, which are used in the following written description.

[0025] The term “Niosome ” refers to a microscopic, self-assembled vesicle used as a drug delivery system for therapeutic applications.

[0026] The present invention discloses a niosomal formulation of bioactive components for topical application. The present invention further discloses aInternal Ref: OR26C021PCT03process of preparation of the niosomal formulation. The niosomal formulation comprises non-ionic surfactants, cholesterol, glycerol, water, bioactive component and propylene glycol. The process of preparation of niosomal formulation comprises steps of mixing cholesterol and non-ionic surfactants, homogenizing the mixture, mixing the polysaccharides, bioactive component, propylene glycol and glycerol, nano milling the mixture, nano milling the homogenized mixture and nano milled mixture to obtain multilamellar niosomes, ultrasonication to obtain large unilamellar niosomes, and ulltrasonication to further obtain small unilamellar niosomes. The obtained niosomes comprising bioactive components are topically administered and does not induce any skin irritation. The niosomal formulation is highly stable.

[0027] Figure 1 tabulates the niosomal formulation of bioactive components for topical application. The formulation comprises non-ionic surfactants at a w / w% concentration range of 2% to 6%. The non-ionic surfactants are selected from tween 60, tween 80 and span 60. The formulation further comprises cholesterol at a w / w% concentration range of 1% to 2%, glycerol at a w / w% concentration of 20%, water as diluent, bioactive component at a w / w% concentration range of 0.5% to 15% and propylene glycol at a w / w% concentration range of 5% to 10%, polysachharide at a w / w% concentration range of 1% to 2%. The bioactive component is selected from vitamin C, vitamin B3 and hyaluronic acid.

[0028] Figure 2 illustrates the flowchart for the process of preparation of niosomal formulation of bioactive component. The process (200) begins with step (201) of preparing a lipid phase by dissolving accurately weighed cholesterol and non-ionic surfactants span 60 and tween 60. The non-ionic surfactants aggregate to form bilayer structures in aqueous environments comprising hydrophobic tail and hydrophilic heads with an aqueous core. The bilayer prevents the coalescence of vesicles and maintains structural integrity. The surfactants do not carry any charge and are more stable, compatible and less toxic. Also, hydrophilic bioactive components are trapped in the aqueous core of the niosomes, while lipophilic bioactive components are incorporated into the bilayer. The ability of non-ionic surfactants to form stable bilayers and accommodate various drug types makes themInternal Ref: OR26C021PCT03ideal for versatile drug delivery applications. Cholesterol fits into the bilayer between the surfactants there by enhancing the rigidity and fluidity of the membrane. Cholesterol further modulates the fluidity of the niosomal membrane. Cholesterol helps in entrapping the bioactive component more effectively hence enhancing the overall drug-loading capacity of the niosomal formulation. At step (202), the mixture of cholesterol and non-ionic surfactants is homogenized to obtain a uniform dispersion. The process of high shear homogenization is carried out at a speed of about 6000 ± 50 rpm and facilitates uniform dispersion of the non-ionic surfactants suspension.

[0029] At step (203), 1% to 2% w / w polysaccharides such as gum ghatti, xanthan gum, guar gum, pectin, etc. are dissolved in water and added to the uniform dispersion. At step (204), aqueous phase is prepared by dissolving glycerol in water at a ratio of 1:1. The active phase is prepared by dissolving bioactive components in propylene glycol and glycerol. At step (205), the obtained active phase of bioactive component and glycerol is subjected to the process of nano milling to obtain a reduced particle size dispersion. The process of nano milling is carried out using zirconia beads having a diameter of about 0.5 ± 0.01 mm and at 6000rpm for a duration of 5 minutes. At step (206), homogenized mixture comprising cholesterol and dispersed suspension of non-ionic surfactants and nano milled mixture comprising bioactive component and glycerol is subjected to mixing. The obtained mixture is subjected to nano milling process to obtain multilamellar niosomes. The process of nano milling is carried out at 6000rpm for a duration of 5 minutes. At step (207), the obtained multilamellar niosomes are subjected to ultrasonication to obtain large unilamellar niosomes. At step (208), the obtained large unilamellar niosomes are further subjected to ultrasonication to obtain nano niosomes.

[0030] The present invention will now be described in detail with reference to certain exemplary embodiments; however, it will be understood that the scope of the invention is not limited to these embodiments.

[0031] Example 1: Preparation of niosomal formulation of vitamin CInternal Ref: OR26C021PCT03

[0032] In an embodiment, the present invention provides a process for the preparation of a vitamin C-loaded nano-niosomal formulation suitable for topical application.

[0033] In an embodiment of the present invention, a lipid phase is prepared by dissolving about 2 w% to about 6 w% of one or more non-ionic surfactants, including Span 60 and Tween 60, in about 1 w% to about 2 w% cholesterol. The resulting mixture is subjected to high-shear homogenization using a homogenizer operated at a speed of about 6000 ± 50 rpm to obtain a substantially uniform dispersion.

[0034] Subsequently, gum ghatti is dissolved in water, separately an aqueous stabilizing phase is prepared by dissolving about 5 w% to about 10 w% propylene glycol in water at a temperature of about 70 ± 0.5 °C until complete dissolution is achieved. The aqueous stabilizing phase is then gradually added to the lipid dispersion under continuous homogenization to form a stable pre-emulsion.

[0035] In parallel, an active phase is prepared by dissolving vitamin C in a glycerol and water mixture, preferably in a ratio of about 1:1, to enhance solubility and stability of the bioactive compound. The pre-emulsion is subjected to a nanomilling process using an agitator bead mill equipped with zirconia beads having a diameter of about 0.5 ± 0.01 mm. The nano-milling is carried out at a rotational speed of about 6000 ± 50 rpm for a duration of about 5 minutes to achieve particle size reduction and improved dispersion. Thereafter, the nano-milled dispersion is combined with the Vitamin C-containing active phase, resulting in the formation of multilamellar niosomes.

[0036] The formulation is further subjected to homogenization at a speed of about 6000 ± 50 rpm for approximately 5 minutes to convert the multilamellar vesicles into unilamellar nano-niosomes having reduced vesicle size and improved uniformity.Internal Ref: OR26C021PCT03

[0037] The resulting vitamin C-loaded nano-niosomal suspension is transferred into light-resistant containers, preferably amber glass vials, sealed with airtight caps, and stored at a temperature of about 4 ± 0.5 °C until further use.

[0038] Example 2: Analysis of ascorbic acid in the nano-niosomal formulation of the present invention

[0039] The amount of ascorbic acid present in the vitamin C-loaded nano-niosomal formulation is determined using a modified analytical method.

[0040] The analysis is carried out using an ultra-performance liquid chromatography (UPLC) system comprising a chromatographic instrument equipped with a Cl 8 column having a particle size of about 5 pm and dimensions of about 4.6 mm x 250 mm. The mobile phase comprises a mixture of acetonitrile and an aqueous buffer solution, preferably 0.05 M ammonium dihydrogen phosphate, adjusted to a pH of about 3 using orthophosphoric acid, in a volumetric ratio of about 90:10.

[0041] The chromatographic separation is performed under isocratic conditions at a flow rate of about 1 mL / min, with the column maintained at a temperature of about 32 °C. Detection of ascorbic acid is carried out using a variable wavelength detector at a wavelength of about 245 nm. The analysis is performed in multiple runs, preferably in triplicate, to ensure precision, reproducibility, and reliability of the obtained results

[0042] The chromatographic method provided a well-resolved and distinct peak corresponding to ascorbic acid at the selected detection wavelength of about 245 nm, indicating the suitability and specificity of the method for analysis. The analysis performed in triplicate demonstrated consistent and reproducible results, with minimal variation between runs, thereby confirming the precision and reliability of the analytical method. The retention time of ascorbic acid remained substantially constant across all runs, indicating stability of the chromatographic system.Internal Ref OR26C021PCT03

[0043] The measured ascorbic acid content confirmed successful incorporation of vitamin C within the nano-niosomal formulation. The high recovery of ascorbic acid from the nano-niosomal formulation further indicates minimal degradation during the preparation process, including nano-milling and homogenization steps.

[0044] Example 3: Determination of Encapsulation Efficiency and Loading Capacity of nano-niosomal vitamin C formulation

[0045] The encapsulation efficiency and the loading capacity of vitamin C in the nano-niosomal formulation of the present invention are determined to evaluate the effectiveness of the encapsulation process.

[0046] A predetermined quantity of the vitamin C-loaded nano-niosomal formulation is dispersed in purified water to obtain a solution of known concentration. The dispersion is subjected to vortex mixing and agitation to ensure uniform distribution of the vesicles. The dispersion is centrifuged at a controlled speed and duration to separate unencapsulated (free) vitamin C from the nano-niosomal vesicles.

[0047] The supernatant containing unencapsulated vitamin C is carefully collected and filtered through a membrane filter, preferably having a pore size of about 0.02 pm, to remove any residual particulate matter. The filtrate is then analyzed using a chromatographic method, such as ultra-performance liquid chromatography (UPLC), at a detection wavelength of about 245 nm to quantify the amount of free Vitamin C.

[0048] The total vitamin C content initially added to the formulation and the amount of unencapsulated Vitamin C present in the filtrate are used to calculate the encapsulation efficiency and loading capacity. Encapsulation Efficiency is determined based on the proportion of vitamin C retained within the niosomal vesicles relative to the total amount added. The loading Capacity is determined based on the amount of vitamin C encapsulated within the vesicles relative to the total weight of the formulation.Internal Ref: OR26C021PCT03

[0049] The results indicated that the niosomal formulation exhibits a high encapsulation efficiency, preferably greater than about 85%, preferably about 90% or higher. In a specific embodiment, the encapsulation efficiency is about 91.87 ± 3.42%, indicating that a substantial portion of vitamin C is successfully entrapped within the niosomal vesicles.

[0050] Further, the loading capacity of the formulation is observed to be about 11.62 ± 1.34%, demonstrating efficient incorporation of the bioactive compound within the vesicular system.

[0051] The high encapsulation efficiency and loading capacity is attributed to the combined effect of non-ionic surfactants and cholesterol forming a stable bilayer structure, which facilitates effective entrapment of hydrophilic vitamin C within the aqueous core. Additionally, the presence of stabilizing agents and the application of nano-milling and homogenization processes contribute to improved vesicle formation and reduced leakage of the encapsulated compound.

[0052] The encapsulation efficiency and loading capacity contribute to enhanced stability, reduced degradation, and improved delivery performance of vitamin C, thereby making the niosomal formulation suitable for pharmaceutical, nutraceutical, and transdermal applications.

[0053] Example 4: Analysis of cytotoxicity of the vitamin C niosomal formulation Using A431 Cells

[0054] The cytotoxicity of vitamin C and vitamin C-loaded niosomal formulation is evaluated using A431 human epidermoid carcinoma cells. The in vitro cytotoxicity of free vitamin C and vitamin C-loaded niosomal formulation was evaluated by an MTT assay following exposure of the cells to varying concentrations of the samples.

[0055] Figure 3 illustrates the cytotoxicity of niosomal vitamin C formulation of the present invention with vitamin C. The results indicated that the vitamin C-loaded niosomal formulation exhibits a lower percentage of cytotoxicity comparedInternal Ref: OR26C021PCT03to free vitamin C across the tested concentration range. The observed cell viability remained comparatively higher in cells treated with the niosomal formulation.

[0056] Further, it was observed that free vitamin C demonstrates minimal cytotoxic effect on A431 cells even at relatively higher concentrations. However, encapsulation of vitamin C within the niosomal vesicles further reduces the cytotoxic impact, indicating improved biocompatibility of the niosomal formulation. The reduced cytotoxicity of the niosomal formulation is attributed to the controlled and sustained release of vitamin C from the vesicular system, thereby preventing sudden exposure of cells to high concentrations of the bioactive compound.

[0057] The cytotoxicity results demonstrate that the vitamin C-loaded nano-niosomal formulation is well tolerated by A431 cells and exhibits enhanced safety profile compared to free vitamin C, thereby supporting its suitability for topical, transdermal, and pharmaceutical applications.

[0058] Example 5: Analysis of In Vitro Permeability of the vitamin C niosomal formulation Using A431 Cells

[0059] The permeability of vitamin C and vitamin C-loaded niosomal formulation is evaluated using A431 human epidermoid carcinoma cells as an in vitro skin model.

[0060] A431 cells are cultured in a suitable growth medium, preferably Dulbecco’s Modified Eagle Medium (DMEM) supplemented with fetal bovine serum and antibiotics, under controlled conditions of about 37 °C and 5% CO2 until a confluent monolayer is formed. The cells are seeded onto permeable supports, such as Transwell inserts, and allowed to differentiate to form a barrier layer mimicking the epidermal membrane. Prior to the study of permeability, the integrity of the cell monolayer is confirmed using transepithelial electrical resistance (TEER) measurements and permeability markers. Only monolayers exhibiting acceptable integrity values are used for further experimentation.Internal Ref: OR26C021PCT03

[0061] A predetermined concentration of free vitamin C solution and an equivalent concentration of vitamin C-loaded nano-niosomal formulation are separately applied to the apical side of the cell monolayer. The basolateral compartment is filled with a suitable receptor medium maintained under physiological conditions. At predetermined time intervals, aliquots are withdrawn from the basolateral compartment and replaced with fresh medium to maintain sink conditions. The collected samples are analyzed for vitamin C content using a suitable analytical technique, preferably ultra-performance liquid chromatography (UPLC), at a detection wavelength of about 245 nm.

[0062] Figure 4 illustrates the permeability of niosomal vitamin C formulation of the present invention with vitamin C. The results indicated that the vitamin C-loaded niosomal formulation exhibits significantly enhanced permeability compared to free vitamin C. The improved permeation is attributed to the nanoscale vesicular structure, which facilitates improved interaction with the cellular membrane and enhanced transport across the epithelial barrier. In addition, the niosomal system provides sustained release of vitamin C, resulting in prolonged permeation over time as compared to the rapid diffusion observed with free vitamin C solution.

[0063] The enhanced permeability and sustained release characteristics contribute to improved bioavailability and efficacy of vitamin C, thereby supporting the suitability of the niosomal formulation for topical, transdermal, and pharmaceutical delivery applications.

[0064] The skin permeation studies revealed that the niosomal formulation exhibited a significantly enhanced permeability profile, with an apparent permeability coefficient approximately 4.77 times higher than that of free vitamin C. This enhancement indicates improved transport of the bioactive compound across the epithelial barrier, thereby supporting its suitability for transdermal delivery applications.

[0065] The formation of stable niosomal formulation is dependent on lipid homogeneity, temperature and particle size. The high shear homogenization processInternal Ref: OR26C021PCT03facilitates breaking down of large aggregates and promotes the formation of small uniform well aligned vesicles which is crucial in creating stable niosomes. Uniform dispersion and reduction in the size of the vesicles aid in enhancing the ability of niosomes to encapsulate and protect the bioactive components. Leakage of bioactive components from the niosomes is hence prevented effectively.

[0066] The encapsulated niosomal matrix fuses with skin cells releasing the bioactive components more effectively with enhanced bioavailability. The well aligned vesicular structure helps to release the bioactive component slowly by facilitating a prolonged release. The niosomal formulation administered topically as a cosmetic, is safe to use and does not induce skin irritation.

[0067] The present invention discloses a niosomal formulation of bioactive component for topical application and a process of preparation thereof. The niosomal formulation and the process of preparation disclosed aids in the overcoming the drawbacks of lack of stability of the niosomes, leakage of bioactive components, skin irritability and so on. The process of preparation of niosomes incorporates optimized temperature facilitating the formation of highly stable niosomes. The formulation of niosomes is administered topically and is bioavailable, safe, effective, skin friendly and stable.

[0068] The niosomal vitamin C formulation provides a synergistic combination of reduced cytotoxicity, enhanced permeability, and improved stability. Accordingly, the formulation represents an effective and delivery system for vitamin C, particularly suited for pharmaceutical, nutraceutical, and cosmeceutical applications.

Claims

Internal Ref: OR26C021PCT03Claims:We Claim,1. A niosomal formulation of bioactive component for topical application, the formulation comprises:a. one or more non-ionic surfactants at a concentration in a range between 2% and 6% w / w;b. cholesterol at a concentration in a range between 1% and 2 % w / w; c. glycerol at a concentration of 20% w / w;d. water as diluent;e. propylene glycol at a concentration in a range between 5% and 10% w / w;f. a bioactive component at a concentration in a range between 0.5% and 15% w / w;g. one or more polysaccharide at a concentration in a range between 1% and 2% w / w;wherein the formulation comprises one or more nano-niosomal vesicles to encapsulate the bioactive component within a bilayer structure in aqueous environment.

2. The niosomal formulation as claimed in claim 1, wherein the bioactive component is selected from vitamin C, vitamin B 12 or hyaluronic acid.

3. The niosomal formulation as claimed in claim 1, wherein the non-ionic surfactants comprise one or more of Span 60, Tween 60, and Tween 80.

4. The niosomal formulation as claimed in claim 1, wherein the polysaccharide is selected from gum ghatti, xanthan gum, guar gum and pectin.

5. The niosomal formulation as claimed in claim 1 , wherein the nano-niosomes are unilamellar vesicles.Internal Ref: OR26C021PCT036. A process for preparation of niosomal formulation of bioactive component, the process (200) comprising the steps of:a. preparing a lipid phase by dissolving 1% to 2% w / w cholesterol and 2% to 6% w / w non-ionic surfactants span 60 and tween 60 (201); b. homogenizing the lipid phase to high shear homogenization to obtain a uniform dispersion (202);c. dissolving 1% to 2 % w / w of one or more polysaccharides in water and adding to the uniform dispersion (203);d. preparing an aqueous phase comprising 20% w / w glycerol in water and preparing active phase by dissolving a 0.5% to 15% w / w bioactive component in a solvent system comprising 5% to 10% w / w propylene glycol and glycerol (204);e. subjecting the active phase to nano milling to obtain a reduced particle size dispersion (205);f. mixing the homogenized lipid phase and polysaccharide mixture with the nano-milled active phase and subjecting the resulting mixture to nano-milling to obtain multilamellar niosomes (206); g. subjecting the multilamellar niosomes to ultrasonication to obtain large unilamellar niosomes (207); andh. subjecting the large unilamellar niosomes to ultrasonication to obtain nano unilamellar niosomes (208).

7. The process as claimed in claim 6, wherein the homogenization of the lipid phase is carried out at a speed of about 6000 ± 50 rpm.

8. The process as claimed in claim 6, wherein the nano milling of active phase is carried out at 6000rpm for a duration of 5 minutes.

9. The process as claimed in claim 6, wherein the nano milling of active phase is carried out using zirconia beads having a diameter of about 0.5 ± 0.01 mm.Internal Ref: OR26C021PCT0310. The process as claimed in claim 6, wherein the ratio of glycerol and water for the aqueous phase is 1:1.

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