DESC:Priority Claim:

[0001] This application claims priority from the provisional application numbered 202041050880 filed with Indian Patent Office, Chennai on 23rd November 2020 entitled “A liposphere formulation for parenteral and topical delivery with improved permeability”, 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 INVENTION
Technical field of the invention
[0003] The present invention relates to a liposphere formulation for delivery of bioconstituents. More particularly, the invention relates to a liposphere formulation for parenteral and topical delivery, with specific application in dermato-cosmetology. The formulation is cost-effective and exhibits improved permeability.
Background of the invention
[0004] Generally, effective delivery system plays a significant role for parenteral and topical drug delivery of bioactive compounds. Liposphere, a lipid microsphere, which is a fat-based encapsulation system, is often used for drug delivery of various bioactive compounds these days. It comprises a lipid-based water dispersible solid particles of particle size between 0.01 and 100 µm in diameter. Typically, a liposphere is composed of a solid hydrophobic lipid core containing active drug moiety dissolved or dispersed in a solid fat matrix, which is stabilized by a layer of phospholipid molecules as external coat.
[0005] Comparatively, liposphere systems are considered to be better than other delivery systems, primarily because the bioactive compound in liposphere is more stable, freeze dry and possess reconstitution properties, controlled particle size, high bioactive compound load, controlled bioactive compound release etc. In addition to this, lipospheres protect the bioactive compound from hydrolysis, improves the shelf life, facilitates high bioavailability and prolonged plasma levels. The lipospheres offer well controlled delivery to a variety of bioactive compounds, such as, anti-inflammatory compounds, local anesthetics, antibiotics, anticancer agents as well as compounds used in dermato-cosmetology.
[0006] In case of dermato-cosmetology or beauty related skin treatments, a wide variety of disorders and diseases affect the skin, e.g., acne, warts, multiple inflammatory dermatoses, skin cancers, autoimmune diseases, occupational dermatoses and contact dermatitis, requiring distinct kinds of investigations and therapies. There are various natural components that are found to be extremely effective to cure such skin disorders and people across the globe prefer using such natural components in order to protect skin from harmful chemicals and to avoid any side-effects.
[0007] The natural components such as curcumin, white curcumin, beta-caryophyllene, alpha lipoic acid, vitamins and ferulic acid, have been found to be extremely effective to cure a range of skin disorders and most of them are administered topically. However, their scarce water solubility is one of the major problems that limits their topical administration. Lots of research in the world has been done in order to develop effective delivery mechanism for these molecules and liposomes, solid lipid nanoparticles, and cyclodextrins have been found to be promising delivery mechanisms.
[0008] The Patent Application No. US20090098168A1 entitled “Multiple-layer microbubble liposphere drug delivery vehicle and system” discloses a multiple layer microbubble drug delivery system, multiple layer microbubble drug delivery vehicle, and a method for fabrication. The microfluidic drug delivery system for producing multiple layer microbubbles includes a first inlet which receives a gas and directs the gas into a central stream, a second inlet which receives a first liquid containing a drug substance and flow focuses the first liquid around the gas and a third inlet which receives a second liquid and flow focuses the second liquid around the first liquid. The multiple layer microbubble drug delivery vehicle includes a gas core, a first liquid layer containing a drug and surrounding the gas core and a second liquid layer surrounding the first liquid layer to stabilize the first liquid layer.
[0009] The Patent Application No. EP3015101B1 entitled “Non-phospholipid liposomes comprising hyaluronic acid” discloses skin care compositions providing lipid vesicles (liposomes) incorporating cross-linked hyaluronic acid. The compositions of the subject invention also comprise vesicles containing collagen. The compositions also contain additional skin care agents. In particular, the invention relates to a non-phospholipid liposome comprising a cavity having encapsulated therein hyaluronic acid, wherein said liposome is about 50 to 950 nm in size, wherein the hyaluronic acid is cross-linked.
[0010] Specifically, with respect to dermato-cosmetology, even though there are various bioactive component delivery mechanisms available today, liposphere formulation, especially for parenteral and topical delivery is popular because it solves one of the major problems of permeability and bioavailability. Even though such formulations have lots of advantages, but the major drawback of these formulations is that they are expensive, and this affects overall price of the final product. Hence, there is a need to have a cost-effective liposphere formulation for parenteral and topical delivery with better efficacy, bioavailability and permeability where such formulation encapsulates various natural cosmetic ingredients and biomolecules to be used in dermato-cosmetology.
[0011] The conventional lipospheres are particulate dispersion of solid spherical particles consisting of solid hydrophobic fat core such as triglycerides or fatty acids derivatives, stabilized by monolayer of phospholipids. The use of phospholipids may be less stable.
[0012] The existing lipospheres are associated with low loading capacity of bioconstituents, may have less stability resulting in the degradation of active molecules and variable kinetics of distribution process.
Summary of the Invention
[0013] In order to overcome the drawbacks of the state of art, the present invention provides a formulation of lipospheres for topical and parenteral delivery of bioactive components. The liposphere formulation comprises glycerolipids such as glyceryl monooleate and glyceryl monostearate at a concentration in the range between 40%-65%, nanofibers of turmeric, ginger or ashwagandha, stearic acid and water.
[0014] Generally, the lipospheres comprises solid lipid core surrounded by a single unit phospholipid layer that entraps the bioactive component or enrich its coat with the bioactive component. The emulsifier or stabilizer is used to form a uniform coat around the core material and to facilitate partitioning of the bioactive component between the lipid and aqueous phases. The lipospheres exhibit high dispersibility in aqueous medium and a release rate for the entrapped substance that is controlled by the phospholipid coating and the carrier.
[0015] The use of plant extracts as nanofibers in the liposphere formulation improves the physical stability and retains the micellar shape of the liposphere matrix. The nanofibers of plant extract used in the invention act as strengthening carriers and stabilize the core liposphere structure thus reinforcing the liposphere vesicles that are strong, stiff and highly crystalline with outstanding thermal stability.
[0016] This liposphere encapsulation ensures maximum bioefficacy, potency and bioavailability for myriad applications in cosmetics by encapsulation with, curcumin, alpha lipoic acid, beta-caryophyllene, vitamins and so on, with the targeted delivery of active ingredients.
[0017] The liposphere of the present invention has been prepared by a solvent evaporation method. This is achieved by evaporation of organic solvent ethanol in which lipids are dissolved allowing the formation of solid microparticles. The process involves dissolution of bioactive molecule in desired quality, glycerolipids, and plant extract in ethanol, followed by evaporation of the solvent at 55°C to 60°C, under reduced pressure condition, using a rotary evaporator. The resulting slurry is high sheared with water for 20-25 minutes with continuous mixing to get a homogenous dispersion.
The lipospheres are useful for parenteral and topical delivery of bioactive molecules especially in dermato-cosmetology and the biomolecules include phytochemicals, herbal extracts, beta-caryophyllene, white curcumin, vitamins and minerals.

[0018] The lipospheres of the present invention are stable, permeable and cost-effective for delivery of the bioactive molecules.
Brief description of the invention
[0019] FIG 1 illustrates the composition of the lipospheres according to an embodiment of the invention.
[0020] FIG 2 illustrates the results of the in vitro release of white curcumin from the liposphere.
[0021] FIG 3 illustrates the results of the in vitro release of ß caryophyllene from the liposphere.
Detailed description of the invention
[0022] 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.
[0023] The term “Bioavailability” refers to the extent and rate at which the active moiety (drug or metabolite) enters systemic circulation, thereby accessing the site of action.
[0024] The term “Dermato-cosmetology” is a scientific super specialty of dermatology.
[0025] The term “Liposphere” refers to fat based encapsulation system developed for parenteral and topical delivery of bioactive compounds.
[0026] The term “Permeability” refers to a parameter which is equal to the diffusion coefficient of the drug or bioactive molecule through the membrane times the membrane/aqueous partition coefficient of the drug or bioactive molecule divided by the membrane thickness.
[0027] The term “Solvent Evaporation Technique” is commonly used method to prepare polymeric nanoparticles, more specifically drug-loaded polymeric systems, generally for pharmaceutical formulations.
[0028] The present invention discloses a novel liposphere formulation, as bioactive component delivery mechanism for parenteral and topical delivery. The liposphere formulation is cost-effective and exhibits improved permeability. The liposphere technology consumes less than the normal dose of the bioconstituents with uncompromised bioavailability.
[0029] The liposphere comprises glycerolipids such as glyceryl monooleate and glyceryl monostearate at a concentration in the range between 40%-65%, nanofibers of turmeric, ginger or ashwagandha, stearic acid and water.
[0030] FIG 1 illustrates the composition of the lipospheres according to an embodiment of the invention. The lipospheres comprises bioactive component at a concentration in the range between 30% to 50%, glycerolipids such as glyceryl monooleate and glyceryl monostearate at a concentration in the range between 40% to 65%, nanofibers from turmeric, ginger or ashwagandha at a concentration in the range between 1% to 5%, stearic acid and water. The use of glycerolipids enhance the core structure of the cosmetic formulation.
[0031] According to an embodiment of the invention, the lipospheres comprises solid lipid core surrounded by a single unit phospholipid layer that entraps the bioactive component or enrich its coat with the bioactive component. The emulsifier or stabilizer is used to form a uniform coat around the core material and to facilitate partitioning of the bioactive component between the lipid and aqueous phases.
[0032] The combination of solid inner core with phospholipid exterior confers several advantages on the lipospheres, including high dispersibility in aqueous medium, and a release rate for the entrapped substance that is controlled by the phospholipid coating and the carrier. In addition, the bioactive component to be delivered does not have to be soluble in the vehicle since it can be dispersed in the solid carrier. The solid core containing a bioactive component dissolved or dispersed in a solid fat matrix and used as carrier for hydrophobic drugs.
[0033] The lipospheres of the present invention utilizes the plant extracts instead of phospholipids as in the conventional lipospheres. The use of plant extracts as nanofibers improves the physical stability and also retains the micellar shape of the liposphere matrix. The plant extracts used in the invention act as strengthening carriers and stabilize the core liposphere structure. These strengthening carriers have been utilized in the invention to reinforce the liposphere vesicles and these resulting vesicles are strong, stiff and highly crystalline with outstanding thermal stability. Such novel lipospheres have been effectively used in dermato-cosmetology by encapsulating with various cosmetic ingredients and biomolecules such as phytochemicals, herbal extracts, beta-caryophyllene, white curcumin, vitamins and minerals.
[0034] Glycerolipids such as glyceryl monooleate and glyceryl monostearate are used in liposphere technology, which enhance the core structure of the cosmetic formulation. Polyglycerol ester of fatty acid, glycerolipids, are a structurally heterogeneous group of lipids that play key structural and functional roles in plant membrane and comprises at least one hydrophobic chain linked to a glycerol backbone in an ester. The conventional liposphere utilizes costly phospholipids to stabilize and to make hydrophilic nature, resulting in overall high price of the final product.
[0035] The liposphere technology disclosed in the present invention uses nanofibers, which contains plant extracts, to stabilize and strengthen the core structure of the liposphere. This novel liposphere offers flexibility to couple with site-specific ligands such as biomolecules to achieve active targeting. This liposphere encapsulation ensures maximum bioefficacy, potency and bioavailability for myriad applications in cosmetics by encapsulation with, curcumin, alpha lipoic acid, beta-caryophyllene, vitamins and so on, with the targeted delivery of active ingredients.
[0036] Several techniques, such as solvent emulsification evaporation, hot and cold homogenization and high-pressure homogenization have been used for the production of lipospheres. Lipid nanoparticle protects the loaded bioactive component from chemical and enzymatic degradation and gradually releases bioactive component from the lipid matrix into blood, resulting in improved therapeutic profiles compared to free drug.
[0037] The use of plant extracts instead of costly phospholipids, as nanofibers to enhance the stability of the liposphere and improve the loading capacity of active biomolecules.
[0038] The liposphere formulation of the present invention has been prepared by a solvent evaporation method. This technique is based on the evaporation of organic solvent in which lipids are dissolved and allowing the formation of solid microparticles. The process involves dissolution of active molecule in desired quality, glycerolipids, and plant extract in ethanol, followed by evaporation of the solvent at 55–60°C, under reduced pressure condition, using a rotary evaporator. The resulting slurry is high sheared with water for 20-25 minutes with continuous mixing to get a homogenous dispersion.
[0039] The lipospheres of the present invention is analysed for release of white curcumin and ß caryophyllene.
[0001] 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: In vitro release of white curcumin from the liposphere
[0002] The liposphere of the present invention is prepared for white curcumin. White curcumin is encapsulated in liposphere and the in vitro solubility and release of white curcumin from liposphere is analyzed. The solubility is analyzed using dimethyl sulfoxide (DMSO) in combination with 7.4 pH phosphate buffer at different concentrations (1:1, 3:7, 1:9, 7: 3, 9:1). 2 grams each of curcumin and white curcumin are separately taken in 50 ml of selected solvent system and visually checked for solubility. The appropriate ratio of the solvent system considered for test for both curcumin liposphere and white curcumin with ethanol and 7.4 pH phosphate buffer in 7:3 ratio.
[0003] In vitro release tests were performed in 7-mL Franz diffusion cells with a diffusion area of 1.86 cm2. The receptor medium comprises phosphate buffer at pH 7.4 which mimics skin conditions, added with DMSO, to ensure sink conditions and to guarantee that the receptor media could not act as a barrier to percutaneous absorption as recommended by FDA guideline (Food and Drug Administration, 1997). The donor compartment contained 200 µL of curcumin liposphere, and the receptor compartment was filled with the respective receptor medium. Hydrophilic polysulfone membrane disc filters were positioned between the cell compartments. The paste was then carefully applied to achieve complete uniform coverage, with the compartments held together using a clamp. Same system was set up with plain white curcumin and considered as control. Stirring rate and temperature were kept at 100 rpm and 32 ± 2 ºC, respectively. 1 mL of aliquots were withdrawn at regular time intervals from 0.25 to 10 hours and collected into HPLC vials, and immediately replaced with the receptor medium at the same temperature. White curcumin liposphere concentrations were correspondingly corrected for replenishments. The withdrawn samples were analyzed to Ultra-High-Performance Liquid Chromatography (UPLC) analysis with mobile phase comprising acetonitrile and 0.1% orthophosphoric acid in a ratio of 32.5:67.5 with C-18 column. Data were expressed as the cumulative amount of the paste permeated through the membrane.
[0004] FIG 2 illustrates the results of the in vitro release of white curcumin from the liposphere. The figure tabulates the release of white curcumin liposphere and white curcumin at different time intervals from 0.25 hours to 48 hours. The results indicated that white curcumin liposphere exhibited a sustained-release profile up to 24 hours, with a total release of 95.6% as shown in Figure. In contrast, the release for white curcumin was much faster, achieving 100.3 % within first hour, indicating the permeability of free white curcumin through the membrane.
Example 2: In vitro release of ß Caryophyllene from the liposphere
[0005] The liposphere of the present invention is prepared for ß caryophyllene. ß caryophyllene is encapsulated in liposphere and the in vitro solubility and release of ß caryophyllene from liposphere is analysed. In vitro release tests were performed in 7-mL Franz diffusion cells with a diffusion area of 1.86 cm2. The receptor medium used is ethanol. The donor compartment contains 200 µL of ß caryophyllene, and the receptor compartment was filled with the respective receptor medium. Hydrophilic polysulfone membrane disc filters were positioned between the cell compartments. The paste was then carefully applied to achieve complete uniform coverage, with the compartments held together using a clamp. Same system was set up with plain ß caryophyllene and was used as control. Stirring rate and temperature were kept at 100 rpm and 32 ± 2 ºC, respectively. 1 mL of aliquots were withdrawn at regular time intervals from 0.25 hours to 48 hours, collected into HPLC vials, and immediately replaced with the receptor medium at the same temperature. ß caryophyllene liposphere concentrations were correspondingly corrected for replenishments. The withdrawn samples were analysed through Gas Chromatography (GC). The data were expressed as the cumulative amount of the paste permeated through the membrane, considering the total amount of drug applied.
[0006] FIG 3 illustrates the results of the in vitro release of ß caryophyllene from the liposphere. The figure tabulates the release of ß caryophyllene liposphere and ß caryophyllene at different time intervals from 0.25 hours to 48 hours. The results indicated that ß caryophyllene exhibited a sustained-release profile up to 24 hours, with a total release of 100.65 % as shown in Figure. In contrast, the release for ß caryophyllene was much faster, achieving 101 % within the first hr, indicating the permeability of free ß caryophyllene through the membrane.
[0007] The novel liposphere formulation is effective as parenteral and topical delivery, specifically in dermato-cosmetology. The formulation is cost-effective and exhibits better bioavailability as well as permeability.

,CLAIMS:Claims:
We Claim.
1. A formulation of liposphere for as bioactive component delivery system, the liposphere comprises:

a. bioactive component at a concentration in a range between 30%-50%;
b. glycerolipids at a concentration in a range between 40%-65%;
c. nanofibers from turmeric, ginger or ashwagandha at a concentration in the range of 1-5%;
wherein the liposphere with a solid lipid core surrounded by a single unit nanofiber layer entraps a bioactive component between the lipid and aqueous phases.
2. The formulation of liposphere as claimed in claim 1, wherein the glycerolipids are selected from glyceryl monooleate or glyceryl monostearate.

3. The formulation of liposphere as claimed in claim 1, wherein the nanofibers from turmeric enhance the stability of the liposphere and improve the loading capacity of bioactive components.

4. The formulation of liposphere as claimed in claim 1, wherein the bioactive components are selected from a group comprising phytochemicals, herbal extracts, beta-caryophyllene, white curcumin, vitamins and minerals.

5. The formulation of liposphere as claimed in claim 1, wherein the liposphere encapsulation improves bio-efficacy, potency and bioavailability of the bioactive components.

6. The formulation of liposphere as claimed in claim 1, wherein the liposphere exhibited a sustained-release profile of white curcumin up to 24 hours with a total release of 95.5%.

7. The formulation of liposphere as claimed in claim 1, the liposphere exhibited a sustained-release profile of ß caryophyllene up to 24 hours, with a total release of 100.65 %.

8. The formulation of liposphere as claimed in claim 1, wherein the liposphere is effective for parenteral and topical delivery of bioactive components.

9. The formulation of liposphere as claimed in claim 1, wherein the liposphere is prepared by solvent evaporation method using ethanol as solvent and the evaporation of the solvent at 55°C to 60°C, under reduced pressure condition.