A formulation of bioactive liposomal gummies and a process of preparation thereof

Abstract

The present invention discloses a formulation of bioactive liposomal gummies and a process of preparation thereof. The formulation comprises a bioactive component, an encapsulating agent, a sweetening agent, a flavoring agent, a gelling agent, an acidity regulator, a coloring agent, a bulking agent, a texturing agent, a thickening agent, a cross-linking polymer and water. The invention discloses a method of preparation of bioactive liposome using nano-milling process subsequently high-pressure homogenization, followed by a spray drying process. The process results in formulating dried bioactive liposomal gummies that exhibit increased stability and bioavailability of the liposomes making it a suitable for drug delivery system for wide range of bioactive agents. The bioactive liposomal gummies exhibit pH and temperature stability along with being palatable.

Description

Internal Ref: OR25C067PCT13TITLE OF THE INVENTIONA formulation of bioactive liposomal gummies and a process of preparation thereofPriority Claim

[0001] This application claims priority from the provisional application numbered 202441059155 filed with the Indian Patent Office, Chennai on 5thAugust 2024 entitled “d formulation of bioactive liposomal gummies and a process of preparation thereof, the entirety of which is expressly incorporated herein by reference.Preamble to the Description

[0002] The following specification particularly 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 formulation of bioactive liposomal gummies and a process of preparation thereof. More specifically, the formulation facilitates the utilization of liposomal technology in delivering palatable and stable gummies with enhanced bioavailability of the encapsulated bioactive molecules.Background of the invention

[0004] The awareness of the necessity of following a healthy diet along with the demand for natural and nutritious food products has increased lately. The development of plant-based and functional gummies has gained the attention of researchers and manufacturers. The development of gummies with alternative gelling agents and sweeteners, natural flavors and colorants, and the incorporationInternal Ref: OR25C067PCT13 of medicines, fiber, protein and antioxidants is of interest. Gummies are a versatile delivery format colorful, varied flavors, and chewable, are easy to consume and palatable for all age groups.

[0005] One of the major drawbacks involved in the gummy matrix is stability and palatability. The preparation method of gummies includes high temperature and low pH. Many of the bioactive components that are either temperature sensitive or pH sensitive will get degraded in these conditions and efficacy of the gummies will be poor. These limitations of the gummy matrix can be easily overcome by incorporating the bioactive components in a stable matrix. One of the most promising technologies which ensures the stability of these sensitive molecules includes liposomal technology.

[0001] Liposomes are small artificial vesicles of spherical shape that are formed from cholesterol and natural non-toxic phospholipids. The vesicle size is an acute parameter in determining the circulation half-life of liposomes, and both size and number of bilayers affect the amount of active molecule encapsulation in the liposomes. Based on the structure, liposomes are classified into four categories based on size and number of bilayers namely small unilamellar vesicles (SUV), large unilamellar vesicles (LUV), multilamellar vesicle (MLV), and multivesicular vesicles (MW). The liposomes are the most popular and promising approach to drug delivery systems compared to numerous drug carriers created to increase the bioavailability and therapeutic efficacy of bio-actives. The liposomes are associated with improved stability, drug delivery, and bioavailability of both hydrophilic and hydrophobic compounds, which are encapsulated or entrapped into various products.

[0002] Further, the liposomal products are mostly available in various forms such as liquid form or suspension form, as the phospholipids are stabilized through a water-in-oil emulsion as a vehicle-like compartment for encapsulation. Although, liposomes exhibit poor stability, due to the random nature of the collapsible bilayer of the phospholipids in the presence of water, leads to a non-uniform development in both size and shape.Internal Ref: OR25C067PCT13

[0003] However, in recent times, the liposomes are also utilized for many active molecules in dietary and nutraceutical industry. Additionally, the liposomes have one or bilayer membranes, and the amphipathic nature of the liposomes is advantageous, as the hydrophilic part is mainly phosphoric acid and bound to a water-soluble molecule, whereas the hydrophobic part consists of two fatty acid chains with 10-24 carbon atoms and 0-6 double bonds in each chain. They form lamellar sheets when dispersed in aqueous medium by aligning themselves in such a way that the polar head group faces outwards the aqueous region while fatty acid groups face each other forming a spherical, vesicle-like structures called as liposomes.

[0004] Further, the liposomes are composed of naturally derived phospholipids with mixed lipid chains, such as egg phosphatidylethanolamine, or of pure surfactant components such as dioleoyl phosphatidylethanolamine (DOP). Generally, the liposomes also contain a core aqueous solution that is trapped by one or more bilayers. The phospholipid bilayers of a liposome originate from natural sources, which are biologically inert, immunogenic and exhibit a lower inherent toxicity.

[0005] The use of liposomes is associated with low stability in aqueous dispersions. Liposomes are associated with chemical and physical degradation eventually resulting in reduced efficacy owing to decreased quality of the formulation.

[0006] In order to overcome the limitations, a suitable liposomal formulation is achieved by choosing an adequate method of preparation. The selection of phospholipids, head group and chain length, as well as the ratio of liposomes components are crucial features to determine safety, stability, and efficiency of liposome.

[0007] In order to reduce the particle size of the molecule in a liquid vehicle, typically aqueous vehicle, the nano milling process is used, wherein the aqueous vehicle is subjected to grinding using polymeric or ceramic media. Nano milling is a proven, commercially validated process for formulating poorly water-solubleInternal Ref OR25C067PCT13 molecules, and the key is the tremendous increase in surface area which directly correlates to an increase in dissolution rate.

[0008] Nano milling is a “top-down” approach of converting large, coarse particles into smaller, finer particles, wherein the process entails the application of mechanical energy to physically break down crystalline structures of molecules. The main technique used for nano milling the molecule is high energy media milling. This technique relies on milling media — 0.2-1 pm beads that are made from ceramics or highly crosslinked polystyrene. The beads shear and collide with the particles during the milling process, reducing their particle size.

[0009] During nano milling, the molecule is suspended in a solution containing water and at least one stabilizer to prevent reaggregation of particles over time. The nano milling process generates an intermediate consisting of nanoparticles suspended in an aqueous vehicle, and it is often referred to as nanoparticulate suspension or nanosuspension, wherein the nanosuspensions have been converted into a wide range of dosage forms, including oral liquids, capsules, tablets, films, injectables, aerosols, and more.

[0010] The homogenization refers to any of several processes used to make a mixture of two mutually non-soluble liquids the same throughout, that is achieved by turning one of the liquids into a state consisting of extremely small particles distributed uniformly throughout the other liquid.

[0011] The Patent Application No. US2018168182A1 entitled “Chewable compositions containing curcuminoids and their method of preparation ” discloses a method for method for the manufacture of gummy composition containing curcuminoids comprising steps of mixing powdered chewable base in water with constant stirring to obtain a turbid solution, adding appropriate concentration of sweeteners to the solution of step a with continuous stirring, mixing the solutions with suitable acidifying agents and flavouring agents to obtain a colloidal mixture, heating the mixture to 55°C.-80° C for 15-20 minutes, adding an effective concentration of curcuminoids to the solution with continuous stirring to form a uniform mixing of the ingredients, pouring the syrup of step e into moulds ofInternal Ref: OR25C067PCT13 different shapes and sizes that is precoated with com starch, allowing the syrup to cool at 10-15° C within the mould and solidify to obtain a chewable gummy, passing the gummies on Vibrosifters to remove excess corn starch, sprinkling suitable sweeteners, acidifying agents and flavouring agents as a coating and packing the solidified chewable candy into suitable containers and packets. The composition of gummy and candy comprises effective concentration of curcuminoids that is palatable and bioavailable.

[0012] The Patent Application No. RU2806858C2 entitled “Chewable tablet and a method for its preparation” discloses a chewable tablet for delivering a bioactive molecule, comprising a compressed core comprising the bioactive molecule, a protective layer covering said compressed core to prevent cracking of said tablet during storage. The protective layer comprises ethyl cellulose, and soft coating layers on top of said protective layer, wherein each soft coating layer contains gum base powder. The protective layer consists of ethylcellulose and a residual organic solvent. The soft coating is covered with a hard coating containing a hard coating binder. The hard coating binder comprises a sugar-based material and gum Arabic.

[0013] The Patent Application No. EP3138558B1 entitled “Liposome composition and production method therefore” discloses a method for preparation of liposomes. The method of adsorbing and retaining a drug onto a lipid membrane of a liposome, release of the drug to the outside of the liposome becomes difficult due to strong interactions such as hydrophobic interactions and electrostatic interactions. Therefore, the configuration of the liposome composition after production can be maintained, whereby it is easy to secure long-term preservation stability.

[0014] In spite of various advantages of the use of liposomes as drug delivery systems, the liposomes are associated with few challenges. One of the drawbacks of liposomes is the rapid degradation due to reticuloendothelial system (RES) and inability to achieve sustained drug delivery over a prolonged period of time. In addition, liposome-based delivery systems are known to possess limitations such as instability, short half-life, and rapid clearance. Also, the manufacturing process ofInternal Ref: OR25C067PCT13 gummies includes pH adjustment and high temperature, resulting in the degradation of many of the sensitive bioactive components, thereby leading to poor efficacy of the gummy.

[0015] Therefore, there is a need for formulation and method for preparation of bioactive liposomal gummies with stable pH and temperature and thus bioavailability of the active molecules.Summary of the invention

[0016] The present invention overcomes the drawbacks of the existing prior art. The present invention discloses a formulation of bioactive liposomal gummies and a process of preparation thereof.

[0017] The present invention discloses a formulation and process of preparation of bioactive liposomes using nano-milling, homogenization followed by spray drying to enhance the bioavailability and stability of the liposomes. The present invention discloses a formulation of bioactive liposomal gummies comprising bioactive components, encapsulating agent, sweetening agents, flavouring agents, gelling agents, acidity regulators, colouring agent, bulking agent, texturing agent, thickening agent, cross linking polymer and water.

[0018] The process results in formulating dried bioactive liposomal gummies that exhibit increased stability and bioavailability of the liposomes making it a suitable drug delivery system for wide range of active agents.Brief description of the drawings

[0019] 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.

[0020] FIG 1 tabulates the formulation of bioactive liposomal gummies, according to an embodiment of the invention.Internal Ref: OR25C067PCT13

[0021] FIG 2a, 2b, 2c illustrates the flowchart disclosing the process of preparation of bioactive liposomal gummies, according to an embodiment of the invention.

[0022] FIG 3 illustrates the schematic representation of the process of preparation of bioactive liposomal gummies.

[0023] FIG 4 illustrates the bioactive liposomal molecule encapsulated in the gummies.

[0024] FIG 5 illustrates the scanning electron microscopic (SEM) visualization of the liposomal gummies.

[0025] FIG 6 illustrates the Transmission Electron Microscopy (TEM) visualization of liposomal gummies.

[0026] FIG 7 illustrates the graphical representation of the zeta potential and particle size distribution of the liposomal gummies.

[0027] FIG 8 illustrates the graphical representation of the release of vitamin C by liposomal gummies and conventional gummies under simulated gastric fluid and simulated intestinal fluid environments.Detailed description of the invention

[0028] 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.

[0029] The term “ Liposomes’" refers to a small artificial vesicle, spherical in shape, having at least one lipid bilayer.

[0030] The term “ Bioavailability” refers to the extent a substance or drug becomes completely available to its intended biological destination.

[0031] The term “Gummy” refers to soft, chewy confectionery made from sugar, flavorings, colorings, and gelling agents.Internal Ref: OR25C067PCT13

[0032] The present invention discloses a formulation and process of preparation of bioactive liposomes using nano-milling, homogenization followed by spray drying to enhance the bioavailability and stability of the liposomes. The present invention discloses a formulation of bioactive liposomal gummies comprising bioactive components, encapsulating agent, sweetening agents, flavouring agents, gelling agents, acidity regulators, colouring agent, bulking agent, texturing agent, thickening agent, cross linking polymer and water. The process results in formulating dried bioactive liposomal gummies that exhibit increased stability and bioavailability of the liposomes making it a suitable drug delivery system for wide range of active agents.

[0033] FIG 1 tabulates the formulation of bioactive liposomal gummies formulation, according to an embodiment of the invention. The bioactive liposomal gummies formulation comprises a bioactive component at a concentration in a range of 150mg to 200mg, an encapsulating agent at a concentration in a range of 80mg to lOOmg, a sweetening agent at a concentration in a range of 900mg to lOOOmg, a sweetening agent at a concentration in a range of 900mg to 950mg, a flavouring agent at a concentration in a range of 50mg to 80mg, a gelling agent at a concentration in a range of 225mg to 275mg, an acidity regulators at a concentration in a range of 75mg to lOOmg, a flavoring agent at a concentration in a range of 25mg to 40mg, a colouring agent at a concentration in a range of 30mg to 50mg, a bulking agent at a concentration in a range of 300mg to 350mg, a texturing agent at a concentration in a range of 50mg to 75mg, a thickening agent at a concentration in a range of O. lmg to 0.5mg, a cross-linking polymer at a concentration range of 75mg to lOOmg and water.

[0034] The bioactive component is selected from a group comprising of vitamin C, multivitamin, vitamin B12, ferrous bisglycinate, magnesium bisglycinate, vitamin D3, curcuminoids, resveratrol, quercetin, ashwagandha, biotin. The bioactive components facilitate enhanced bioavailability and protection, leading to improved effectiveness.Internal Ref: OR25C067PCT13

[0035] The encapsulating agent incorporated is lecithin. The encapsulating agent aids in protecting and delivering bioactive components. The encapsulating agent aids in the formation of liposomes and in encapsulating the active compounds. The encapsulating agent shields from degradation of the bioactive component by stomach acid and enzymes hence enhances the stability and bioavailability of the active ingredients, ensuring they reach the bloodstream and target tissues more effectively.

[0036] The sweetening agents incorporated are sorbitol syrup and maltitol. The flavoring agent incorporated is orange powder. The gelling agent incorporated is pectin. The gelling agent provides the desired texture and structure to the formulation, transforming a liquid liposome into a solid, chewable form The acidity regulators incorporated are citric acid and sodium citrate. The flavoring agent incorporated is natural orange. The coloring agent incorporated is natural color.

[0037] The bulking agent incorporated is gum Arabic. Gum Arabic helps to maintain the stability of the liposomes, preventing them from aggregating or separating out, and also contributes to a uniform dispersion of the liposomes within the gummy matrix. Furthermore, gum Arabic enhances overall structure and texture of the gummy. The texturing agent incorporated is olive oil. The thickening agent incorporated is carnauba wax and water as diluent. Texturing agent and thickening agent facilitates in gelling process and influences the optimal firmness, chewiness, mouthfeel and palatability of the liposomal gummy.

[0038] The cross-linking polymer incorporated is selected from a group of trehalose, xyloglucan and citric acid. The addition of cross-linking aids in stabilizing the liposomal structure encapsulating the bioactive components. Additionally, the cross-linking polymer enhances the overall stability and release profile of the gummies. Cross-linking enhances the mechanical properties of the liposomal network and enhances the resistance to degradation and further improves the ability to deliver the bioactive components effectively. Also, cross-linking polymer creates a network within the liposome, aiding in prevention of leakage of the encapsulated bioactive components.Internal Ref: OR25C067PCT13

[0039] FIG 2a, 2b, 2c illustrates the flowchart disclosing the process of preparation of bioactive liposomal gummies, according to an embodiment of the invention. The process of preparation of bioactive liposomal gummies is segregated into processes 200a, 200b & 200c. FIG 2a illustrates the flowchart for the process (200a) comprising steps (201a) where pectin is dissolved in boiling water at a concentration in a range of 225mg to 275mg. At step (202a), pectin dissolved in water is subjected to de-aeration. Further, at step (203a), the sweetening agent at a concentration range of 900mg to lOOOmg is dissolved in water, and at step (204a), the sweetening agent is dissolved in water and subjected to boiling followed by cooling process. Additionally, at step (205a), the solution of de-aerated pectin solution and the sweetening agent are subjected to mixing.

[0040] FIG 2b illustrates the flowchart for the process (200b) comprising steps of: (201b) where bioactive molecules at a concentration in a range of 150mg to 250mg are dissolved in water. Further, at step (202b), gum Arabic at a concentration range of 300mg to 350mg is mixed with the solution of bioactive molecules. Additionally, at step (203b), the solution mixture is subjected to the process of nano milling. The mixture subjected to nano milling undergoes particle size reduction wherein the chamber capacity is 80ml with a bead size of 0.5mm and 3.7 kg / m3 density per beads. The rotation conditions incorporated are 6000rpm for a duration of 10 to 25 minutes. The process of nano milling aims at particle size reduction for enhancing the solubility, absorption and hence bioavailability of the bioactive molecule.

[0041] Further, at step (204b), the phospholipids at a concentration of 80mg-100mg is dissolved in water at a temperature of 80°C, and the obtained mixture is subjected to nano milling. The mixture subjected to nano milling undergoes particle size reduction wherein the chamber capacity is 80ml with a bead size of 0.5mm and 3.7 kg / m3 density per beads. The rotation conditions incorporated are 6000rpm for a duration of 10 to 25 minutes. The process of nano milling aims at particle size reduction for enhancing the solubility, absorption and hence bioavailability of the bioactive molecule.Internal Ref: OR25C067PCT13

[0042] Further, at step (205b), the obtained mixture is subjected to homogenization under high pressure conditions, and subsequently the process of homogenization under high pressure conditions is repeated 3 times, in step (206b). The homogenization is conducted at a pressure of 1000 bars and 10,000rpm. The pressure and rotational speed are optimized in order to maintain uniform particle size distribution upon size reduction. Further, at step (207b), the homogenized mixture is subjected to spray drying in order to obtain the dry bioactive liposomal in powdered form. The spray drying parameters involve an inlet temperature of 90°C and an outlet temperature of 170°C, wherein the inlet temperature affects drying rate and thermal efficiency, while the outlet temperature influences moisture content and particle characteristics.

[0043] FIG 2c illustrates the flowchart for the process (200c) comprising steps (201c) wherein the dry liposomal powder and mixture of cross-linking polymers at a concentration range of 75mg to lOOmg are subjected to mixing. Further at step (202c), the matrix coated dry liposomal powder and mixture of sweetening agent and pectin are subjected to mixing. Further, at step (203c), the acidity regulator at a concentration range of 75mg to lOOmg, the natural color at a concentration range of 30mg to 50mg and the flavor at a concentration range of 25mg to 40mg are incorporated into the mixture. Furthermore, at step (204c), the thickening agent at a concentration range of O.lmg to 0.5mg, the texturing agent at a concentration range of 50mg to 75mg are incorporated into the mixture. At step (205c), the obtained mixture is subjected to cooling, wherein the cooled mixture is poured into a mold to obtain a bioactive liposomal gummy in step (206c).

[0044] FIG 3 illustrates the schematic representation of the process of preparation of bioactive liposomal gummies. The schematic representation illustrates the mixing of the solution comprising gum Arabic and water along with the solution of bioactive components at step (i). Further, under controlled temperature by the process of soft nano milling, the phospholipids are added to the mixture comprising gum Arabic and bioactive components at step (ii). Subsequently, the mixture is homogenized at high pressure conditions of 1000 bars and temperature controlled soft milling process at step (iii). The obtained liposomal mixture is subjected toInternal Ref: OR25C067PCT13 spray drying to obtain bioactive liposomes at steps (iv) & (v).

[0045] FIG 4 illustrates the bioactive liposomal molecule encapsulated in the gummies. The bioactive liposome comprises bioactive molecule depicted as (a) dispersed in water depicted as (b) along with gum Arabic depicted as (c) in the center surrounded by a bilayer of phospholipids depicted as (d). The process aids the preparation of hydrophilic as well as hydrophobic bioactive liposomal gummies as the bioactive components are well protected inside the liposomal matrix hence preventing the degradation of the bioactive components, hence increasing the bioavailability.

[0046] The process disclosed in the present invention facilitates the formation of stable liposomal gummies with encapsulated bioactive components with enhanced stability. The formulation of bioactive liposomal gummies with exhibits sustained release upon administration. Furthermore, the formulation of bioactive gummies facilitates uniform particle size and distribution.

[0047] The following examples are offered to illustrate various aspects of the invention. However, the examples are not intended to limit or define the scope of the invention in any manner.Example 1: Characterization of bioactive liposomal gummies with Vitamin C

[0048] The characterization studies were evaluated for liposomal gummies comprising vitamin C as the bioactive component, wherein the characterization studies are crucial in determining and analyzing the physical properties of the gummies. The analysis of physical properties of the liposomal gummies aids in understanding the drug delivery upon administration. The characterization studies further facilitate in the process optimization to enhance the efficacy of the formulation obtained. Additionally, the characterization studies also illustrate the molecular behavior of the formulation.

[0049] According to an embodiment of the invention, the formulation of liposomal gummies with vitamin C as bioactive component was evaluated to analyze theInternal Ref: OR25C067PCT13 liposomal structure by scanning electron microscopy (SEM), Transmission electron microscopy (TEM), particle size distribution, zeta potential and simulated gastrointestinal digestion.

[0050] The formulation of liposomal gummies with vitamin C as bioactive component was visualized under SEM. The formulation was sampled and stored in double-sided carbon ribbon- wrapped aluminium stubs, sputter-coated with a thin layer of gold by a sputter gold coater. The sampled formulation was subjected to scanning at an accelerating voltage of 10 kV.

[0051] FIG 5 illustrates the scanning electron microscopic (SEM) visualization of the liposomal gummies. The illustration depicts the morphology of the liposomal structure incorporated in the gummies. The liposomal gummies exhibit a well dispersed, spherical shape with fold-like surface indicating the formation of crosslinked polymeric matrix coating with gummy layers over the liposomal vitamin C formulation without modifying the characteristic liposomal structure.

[0052] The morphology of the liposomal gummies was further analyzed by TEM. The formulation was sampled and dispersed by placing it in a sonicator for a duration of 10 minutes, wherein the sample comprising the formulation was subjected to disperse on glow-discharged thin carbon coated TEM micro grids, and the dispersed sample was allowed to dry at room temperature.

[0053] FIG 6 illustrates the graphical representation of Transmission Electron Microscopy (TEM) visualization of liposomal gummies. The illustration depicts distinct dark interface inside attributed to the well encapsulation of vitamin C with self-emulsified nanospheres. The nanospheres were well surrounded by transparent phospholipid layer. Furthermore, a dark colored coating around the surface of phospholipoid layer exhibits successful coating of crosslinked polymeric matrix in the formulation of liposomal gummies.

[0054] The formulation of liposomal gummies was analyzed for zeta potential and particle size distribution. Zeta potential and particle size distribution analysis are crucial for understanding and optimizing lipid-based drug delivery systems.Internal Ref OR25C067PCT13Zeta potential reflects the surface charge of the particles, colloidal stability and interactions. Particle size distribution impacts drug encapsulation, release, and biodistribution.

[0055] The dynamic light scattering (DLS) method was employed to determine the mean particle sizes and zeta potentials of the bioactive liposomal formulation by DLS-nanoZS, Zetasizer Nanoseries and Malvern Instruments. The formulation was dispersed and subjected to dilution with water. The zeta potential was measured at a temperature of 25 °C. The particle sizes of the formulation of liposomal gummies were determined using the Stokes-Einstein equation. The zeta potentials were determined using the Nano DTS software. The measurement was executed at least three sets of six runs.

[0056] The zeta potential aids in analyzing the probability of particle aggregation. Higher zeta potential indicates enhanced stability. FIG 7 illustrates the graphical representation of the zeta potential and particle size distribution of the liposomal gummies. The zeta potential of the liposomal gummies was found to be -31.98 mV. The zeta potential indicates a homogenous distribution of the liposomal gummies without aggregation and are well dispersed.

[0057] Further, mean particle size of the liposomal gummies was analyzed. Mean particle size influences physical stability, chemical stability, solubility, turbidity, release rate and biological performance. The liposomal gummies exhibited particle size range of 170 nm to 320 nm with an average particle size as 247.5±22.4 nm. Moreover, the results clearly revealed that the sizes of the particles exist in a narrow range and are uniform.Example 2: In vitro analysis of stability and sustained release of the bioactive liposomal gummies with Vitamin C

[0058] The formulation of liposomal gummies with vitamin C as bioactive component was subjected to in vitro analysis of stability and sustained release in comparison to conventional vitamin C. The formulation of liposomal gummies and conventional vitamin C were subjected to Simulated Gastric Fluid (SGF) andInternal Ref: OR25C067PCT13Simulated Intestinal Fluid (SIF) environments for a duration of 12 hours. The rate of vitamin C release in SGF and SIF from liposomal gummies and conventional gummies with an initial weight of 2.8 g were incubated in 900 ml of SGF at a pH of 1.2 and SIF at pH of 6.8 solutions with a stirring speed at 120 rpm for a period of 12 hours at a temperature of 37 °C. The liposomal gummies and conventional gummies were subjected to filtration by placing them within a filtration bag and immersed in a beaker including 900 ml of SGF or SIF.

[0059] FIG 8 illustrates the graphical representation of the release of vitamin C by liposomal gummies and conventional gummies under simulated gastric fluid and simulated intestinal fluid environments. The illustration depicts a burst release of vitamin C at a percentage of 58% of vitamin C from conventional gummies occurred after a duration of 30 minutes. Subsequently, the release of vitamin C reached 67% after a duration of 2 hours following which reaching to 72% at a duration of 12 hours. The liposomal gummies exhibited slow release at a percentage of 22% of vitamin C up to a duration of 2 hours. The decrease in rate of release is due to the well encapsulation of vitamin C in the liposomal core of the gummies with surface coated by the cross-linked polymeric coating. The sustained release indicates the surface coated liposomes could resist the destruction of gastric ph.

[0060] Under SIF environment, the conventional gummies exhibited a burst release with an initial release at a percentage of 61% up to a duration of 1 hour and subsequently release steadily decreased up to a percentage of 43% up to a duration of 12 hours. The liposomal gummies exhibited enhanced release. The cumulative release rate of liposomal gummies after a duration of 2 hours under the SIF environment was at a percentage of 64% and exhibited an increase in release at a percentage of 78% up to a duration of 12 hours. The encapsulation of vitamin C in the liposomal gummies exhibited higher vitamin C release compared to conventional gummies under SIF environment. The higher vitamin C release depicts stabilization of vitamin C.

[0061] The characterization and in vitro studies indicate that the formulation of liposomal gummies was nanosized, spherical, without any aggregation, illustratingInternal Ref: OR25C067PCT13 the stability of the formulation. The evaluation was further supported by TEM analysis. The formulation of liposomal gummies exhibit an optimal particle size range of 170 nm to 320 nm with an average particle size as 247.5±22.4 nm. The particles exhibit homogenous distribution, well dispersed with good stability, zeta potential -31.98 mV. The bioactive component is well protected in the liposomal structure.

[0062] In addition, the liposomal gummies exhibited improved release of vitamin C due to improved encapsulation of vitamin C in the liposomal formulation.. Accordingly, liposomal encapsulation of vitamin C followed by coating through crosslinked polymeric matrix into the preparation of aids in retaining the potency of the bioactive component.

[0063] The present invention discloses a formulation of bioactive liposomal gummies and a process of preparation thereof. The bioactive liposomal gummies exhibit pH and temperature stability along with being palatable. The formulation of bioactive liposomal gummies prepared by said method are stable, results in controlled release and enhances the absorption and bioavailability of the encapsulated bioactive molecules. The bioactive liposomal gummies of the present invention are useful for encapsulation of various ranges of active molecules. The bioactive liposomal gummies exhibit improved stability, extended half-life, and delays the degradation as a result of spray drying.

Claims

Internal Ref: OR25C067PCT13Claims:We Claim:

1. A formulation of bioactive liposomal gummies, the formulation comprising: a) a bioactive component at a concentration range of 150mg to 200mg; b) an encapsulating agent at a concentration range of 80mg to lOOmg; c) a sweetening agent at a concentration range of 900mg to lOOOmg; d) a sweetening agent at a concentration range of 900mg to 950mg; e) a flavoring agent at a concentration range of 50mg to 80mg, f) a gelling agent at a concentration range of 225mg to 275mg; g) one or more acidity regulators at a concentration range of 75mg to lOOmg; h) a flavouring agent at a concentration range of 25mg to 40mg; i) a coloring agent at a concentration range of 30mg to 50mg; j) a bulking agent at a concentration range of 300mg to 350mg; k) a texturing agent at a concentration range of 50mg to 75mg; l) a thickening agent at a concentration range of 0. lmg-0.5mg; m) a cross-linking polymer at a concentration range of 75mg to lOOmg; and n) water. wherein the components are combined to form bioactive liposomal gummies formulation with enhanced palatability, stability, absorption and bioavailability.Internal Ref: OR25C067PCT132. The formulation as claimed in claim 1, wherein the bioactive component is selected from a group comprising of vitamin C, multivitamin, vitamin B 12, ferrous bisglycinate, magnesium bisglycinate, vitamin D3, curcuminoids, resveratrol, quercetin, ashwagandha, biotin.

3. The formulation as claimed in claim 1, wherein the encapsulating agent is lecithin, the sweetening agents selected from a group of sorbitol syrup and malitol, the flavoring agent is orange powder, the gelling agent is pectin, the acidity regulator is selected from a group of citric acid and sodium citrate, the flavouring agent is natural orange, the coloring agent is natural color, the bulking agent is gum Arabic, the texturing agent is olive oil, the cross linking polymer is selected from a group of trehalose, xyloglucan and citric acid and the thickening agent is carnauba wax.

4. The method for the preparation of the bioactive liposomal gummies, the method (200) comprises steps of: a) dissolving pectin at a concentration range of 225mg to 275mg in boiling water; (201a) b) subjecting pectin dissolved in water to de-aeration; (202a) c) dissolving a sweetening agent at a concentration range of 900mg tolOOOmg in water; (203a) d) subj ecting the mixture of sweetening agent dissolved in water to boiling followed by cooling; (204a) e) combining the de-aerated pectin solution and the solution of sweetening agent to form a uniform mixture; (205a) f) dissolving bioactive molecules at a concentration of 150mg to 250mg in water; (201b) g) mixing gum Arabic at a concentration range of 300mg to350mg with the solution containing the bioactive molecules; (202b) h) subjecting the solution mixture to nano milling for particle size reduction; (203b)Internal Ref: OR25C067PCT13 i) dissolving phospholipids at a concentration of 80mg tolOOmg in water and subjecting the obtained mixture to nanomilling; (204b) j) subjecting the obtained mixture to homogenization under high pressure for particle size reduction and uniform distribution; (205b) k) repeating the process of homogenization 3 times; (206b) l) subjecting the homogenized mixture to spray drying to obtain dry bioactive liposomal powder; (207b) m) subjecting the dry liposomal powder and mixture of cross-linking polymers at a concentration range of 75mg to lOOmg to mixing; (201c) n) subjecting the matrix coated dry liposomal powder and mixture of sweetening agent and pectin to mixing; (202c); o) adding acidity regulators at a concentration range of 75mg to lOOmg, natural colors at a concentration range of 30mg-50mg and flavours at a concentration range of 25mg to40mg to the mixture; (203c) p) adding thickening agents at a concentration range of O. lmg to0.5mg, texturing agents at a concentration range of 50mg to75mg to the mixture; (204c); q) subjecting the obtained mixture to cooling at room temperature; (205c) and r) pouring the cooled mixture into molds to obtain bioactive liposomal gummies. (206c) wherein, the process facilitates the formation of well dispersed, spherical liposomal gummies with uniform particle size encapsulating the bioactive component and facilitates sustained release with enhanced bioavailability.

5. The method as claimed in claim 4, wherein the homogenization is carried out at a pressure of 1000 bars and 10,000 rpm for 3 cycles for particle size reduction and uniform particle size distribution.

6. The method as claimed in claim 4, wherein nanomilling for particle size reduction is carried out at a chamber capacity of 80ml, ahead size of 0.5mm,Internal Ref: OR25C067PCT13 a density of 3.7 kg / m3 per bead, rotating at a speed of 6000rpm for a duration of 10 minutes to 25 minutes.

7. The method as claimed in claim 4, wherein the inlet temperature for spray drying is 90°C and the outlet temperature is 170°C.

8. The method as claimed in claim 4, wherein the particle size of the liposomes is in a range of 170nm to 320 nm.

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