KR102072439B1 - Method for preparing liposome by environment-friendly solvent - Google Patents

Abstract

The present invention relates to a method of dispersing phospholipids, which are components of liposomes, by using alcohol, which is a solvent for food, and preparing the same by using a high pressure emulsion. Unlike the basic method of preparing liposomes using toxic organic solvents such as hexane, ethyl acetate and chloroform, the present invention can be directly applied to the food industry by preparing liposomes using non-toxic spirits used in food manufacturing. By using high pressure emulsifier, mass production is possible. In addition, in the past, liposome powder was prepared by lyophilization using a sugar component and an additional solvent, which was inefficient in terms of time and cost. However, the present invention simplifies the complicated and complicated powder manufacturing process with a concentration process and replaces it by spray drying. Has the ease of minimization and cost reduction.

Description

Liposome manufacturing method using eco-friendly solvent {METHOD FOR PREPARING LIPOSOME BY ENVIRONMENT-FRIENDLY SOLVENT}

The present invention relates to a method of dispersing phospholipids, which are components of liposomes, by using alcohol, which is a solvent for food, and preparing the same by using a high pressure emulsion. In particular, in order to solve the shortcomings of the manufacturing method using organic solvents that are limited in use in existing foods, and to increase the ease of industrial application, by establishing a powdered manufacturing process through mass and spray drying techniques, food, cosmetics, It is a manufacturing technology that can be widely used in the pharmaceutical industry.

Nano-encapsulation technology is widely used to protect active ingredients from the external environment and increase bioavailability, and liposomes are double-layered nanocarriers that can be manufactured by the self-assembly principle of emulsifiers. (nano-carrier, vesicle). These liposomes can be loaded with both fat-soluble and water-soluble physiologically active substances as well as reduce the toxicity of the loaded drugs and protect the active drugs from external factors such as heat, light and enzymes. It is evaluated to have high availability as a carrier in a system.

Liposomes have the advantage of increasing the bioavailability of poorly soluble materials and are highly available. However, liposomes are used as foods because they use toxic organic solvents as solvents to disperse emulsifiers, which are components of liposomes. In order to do so, the KFDA requires certification. Due to these shortcomings, toxicological evaluation should be carried out in order to use in various industries, and toxic organic solvents are inconvenient to be used only for a specific purpose (extraction solvent) and are absolutely impossible to use as food. However, it is known that pharmaceuticals are less regulated than foods, and are manufactured by using toxic organic solvents and commercializing them. While liposomes are highly available due to the above-mentioned advantages, in order to disperse emulsifiers as constituents of liposomes, toxic organic solvents are used as solvents. However, due to the above disadvantages, it is difficult to apply to the food industry, there is an inconvenience that the toxicity evaluation should proceed in order to use the material in various industries.

In addition, liposomes have the disadvantage of chemical decomposition or physical change during the manufacturing process or storage. Chemical degradation is caused by the oxidation and hydrolysis of the phospholipid (lecithin), an emulsifier, and physical changes include agglomeration, fusion, and leakage of active ingredients in liposome particles in aqueous solution. Therefore, a method for stably storing liposomes from these factors is needed.

Existing previous studies have reported a number of research results of powdering liposomes through lyophilization. In the preceding studies, additional sugar components (trehalose, dextrin, sucrose, etc.) are added during lyophilization to contain high concentrations of sugar, and solvents are used to control the rate of constant freezing. Specifically, the previous studies reported above use a toxic organic solvent in liposome manufacturing process and apply powdering technique in several steps, but it is known that it must contain an additional sugar component and can be additionally applied in various processes. . As a result, it is impossible to apply economically and industrially immediately, and the market competitiveness due to high production cost is low.

Therefore, in order to overcome these disadvantages, the present inventors have completed the present invention by conducting a study on the powdering process of liposomes and non-toxic solvents that can disperse the phospholipids of the liposomes.

KR 10-2015-0074390 A

In one aspect, an object of the present invention is to prepare a micelle by mixing the dispersion and the primary hydrophilic solution; Preparing a multiphase emulsion by mixing a second hydrophilic solution and the prepared micelle; Preparing a liposome solution; And it provides a liposome manufacturing method comprising the step of preparing a liposome powder.

In another aspect, an object of the present invention is to provide a liposome prepared by the liposome production method.

In one aspect, the present invention comprises the steps of preparing a micelle (mixelle) by mixing a dispersion in which phospholipids are dispersed in alcohol and a primary hydrophilic solution in which the primary hydrophilic active material is dissolved in a primary hydrophilic solvent; Preparing a multiple emulsion by mixing a second hydrophilic solution in which a second hydrophilic active material is dissolved in a second hydrophilic solvent and the prepared micelle; Preparing a liposome solution by removing alcohol from the prepared multiphase emulsion using a concentrator; And spray drying the liposome solution to prepare a liposome powder.

In another aspect, the present invention provides a liposome prepared by the liposome production method.

Unlike the basic method of preparing liposomes using toxic organic solvents such as hexane, ethyl acetate and chloroform, the present invention can be directly applied to the food industry by preparing liposomes using non-toxic spirits used in food manufacturing. By using high pressure emulsifier, mass production is possible. In addition, in the past, liposome powder was prepared by lyophilization using a sugar component and an additional solvent, but it was inefficient in terms of time and cost. Has the ease of minimization and cost reduction.

1 is a schematic diagram of a liposome manufacturing process of the present invention. The hydrophilic core material of FIG. 1 is the hydrophilic active material of one embodiment of the present invention.
FIG. 2 is a distribution diagram of the liposome particle size of the liposome solution of the present invention using dynamic light scattering (DLS).
3 is a distribution chart of the liposome particle size of the liposome powder of the present invention confirmed by dynamic light scattering (DLS).
Figure 4 is a photograph of the liposome powder of the present invention.
5 is a photo of the liposome powder of the present invention taken with a transmission electron microscope (TEM).

Hereinafter, the present invention will be described in detail.

In one aspect, the present invention comprises the steps of preparing a micelle (mixelle) by mixing a dispersion in which phospholipids are dispersed in alcohol and a primary hydrophilic solution in which the primary hydrophilic active material is dissolved in a primary hydrophilic solvent; Preparing a multiple emulsion by mixing a second hydrophilic solution in which a second hydrophilic active material is dissolved in a second hydrophilic solvent and the prepared micelle; Preparing a liposome solution by removing alcohol from the prepared multiphase emulsion using a concentrator; And spray drying the liposome solution to prepare a liposome powder.

The liposomes of the present invention are spherical vesicles in which the phospholipid bilayer surrounds the water phase, and are mostly spherical or elliptical. Since there is a space inside the liposome, it is used to deliver active substances such as polymers, drugs, and antigens to the body.

Phospholipids of the present invention may be dispersed in an organic solvent and then mixed with a solution containing an active substance to form micelles. The phospholipid may be natural phospholipid or synthetic phospholipid. Specifically, the phospholipid is lecithin, phosphatidylcholine (PC), phosphatidylglycerol (PG), phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidic acid (PA), phosphatidyl inositol (PI), egg phosphatidylcholine (EPC), egg phosphatidylglycerol (EPG), egg phosphatidylethanolamine (EPE), egg phosphatidylserine (EPS), egg phosphatidyl acid (EPA), egg phosphatidyl inositol (EPI), soy phosphatidylcholine (SPC), soy phosphatidylglycerol (SPG), Soy phosphatidylethanolamine (SPE), Soy phosphatidylserine (SPS), Soy phosphatidyl acid (SPA), Soy phosphatidyl inositol (SPI), dipalmitoylphosphatidylcholine (DPPC), 1,2-dioleoyl- sn-glycero-3-phosphatidylcholine (DOPC), dimyristoylphosphatidylcholine (DMPC), dipalmitoylphosphatidylglycerol (DPPG), diol phosphatidylglycerol (DOPG), dimyristoylphosphatidylglycerol (DMPG), Sardecylphosphocholine (HEPC), hydrogenated soybean phosphatidylcholine (HSPC), distearoylphosphatidylcholine (DSPC), distearoylphosphatidylglycerol (DSPG), dioleylphosphatidylethanolamine (DOPE), palmitoylstearo Ilphosphatidylcholine (PSPC), palmitoylstearoylphosphatidylglycerol (PSPG), monooleoylphosphatidylethanolamine (MOPE), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC), polyethylene Glycol distearoylphosphatidylethanolamine (PEG-DSPE), dipalmitoylphosphatidylserine (DPPS), 1,2-dioleoyl-sn-glycero-3-phosphatidylserine (DOPS), dimyristoylphosphatidylserine (DMPS), distearoylphosphatidylserine (DSPS), dipalmitoylphosphatidic acid (DPPA), 1,2-dioleoyl-sn-glycero-3-phosphatidic acid (DOPA), dimyristoyl Phosphatidic acid (DMPA), distearoylphosphatidic acid (DSPA), Dipalmitoylphosphatidyl inositol (DPPI), 1,2-dioleoyl-sn-glycero-3-phosphatidyl inositol (DOPI), dimyristoyl phosphatidyl inositol (DMPI) or distearoyl phosphatidyl inositol (DSPI) yl And, more specifically, it may be lecithin, but is not limited thereto.

Alcohol of the present invention is ethanol, a food alcohol, is non-toxic and can be used in food production. The spirit of the present invention may be 90 to 100% alcohol, specifically 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more Or 99% or more of alcohol, 100% or less, 99% or less, 98% or less, 97% or less, 96% or less, 95% or less, 94% or less, 93% or less, 92% or less or 91% or less In more detail, it may be 95% of alcohol, but is not limited thereto.

The dispersion of the present invention is a phospholipid dispersed in alcohol, the phospholipid may be included in 0.1 to 1% by weight relative to the total weight of the dispersion. Specifically, the phospholipid may be 0.1 wt% or more, 0.2 wt% or more, 0.3 wt% or more, 0.4 wt% or more, 0.5 wt% or more, 0.6 wt% or more, 0.7 wt% or more, 0.8 wt% or more, based on the total weight of the dispersion, or 0.9 wt% or more, 1 wt% or less, 0.9 wt% or less, 0.8 wt% or less, 0.7 wt% or less, 0.6 wt% or less, 0.5 wt% or less, 0.4 wt% or less, 0.3 wt% or less, 0.2 wt% or less It may be included in% or less or 0.1% by weight or less, more specifically 0.5% by weight, but is not limited thereto.

According to an embodiment of the present invention, as a result of research on a nontoxic solvent capable of dispersing phospholipids, which are components of liposomes, in order to disperse phospholipids well, phospholipids are uniformly dispersed by using alcohol, and even particles are selected. In order to make the dispersion was prepared using a high pressure emulsifying device during the dispersion process (Example).

The dispersion may further disperse the sterols in alcohol. The sterol is an alcohol having a steroid skeleton widely distributed in the flora and fauna, animal-derived sterols include cholesterol, coprostanol, and the like, plant-derived sterols include cytosterol, and fungus-derived sterols include ergosterol. . The sterol may be cytosterol, and more specifically β-sitosterol, but is not limited thereto. The sterol may be included in an amount of 0.01 to 0.1 wt% based on the total weight of the dispersion. Specifically, the sterol is 0.01% by weight, 0.02% by weight, 0.03% by weight, 0.04% by weight, 0.05% by weight, 0.06% by weight, 0.07% by weight, 0.08% by weight or more based on the total weight of the dispersion Or 0.09 wt% or more, and 0.1 wt% or less, 0.09 wt% or less, 0.08 wt% or less, 0.07 wt% or less, 0.06 wt% or less, 0.05 wt% or less, 0.04 wt% or less, 0.03 wt% or less, 0.02 It may include less than or equal to 0.01% by weight, more specifically 0.0625% by weight, but is not limited thereto.

Primary hydrophilic active substances of the invention include branched amino acids, hydrophilic drugs and the like. The branched amino acid is an amino acid having a branched alkyl group in the side chain, and specifically, may be valine, leucine, or isoleucine, but is not limited thereto. The hydrophilic drug may be, but is not limited to, a cell membrane dye, a photosensitizer, an anticancer agent, or an antibiotic.

The primary hydrophilic solvent of the present invention may be specifically water, but is not limited thereto.

The primary hydrophilic solution of the present invention is to dissolve the primary hydrophilic active material in the primary hydrophilic solvent, the primary hydrophilic active material may be included in 0.0001 to 99% by weight relative to the total weight of the primary hydrophilic solution. Specifically, the primary hydrophilic active material is 0.0001% by weight, 0.0005% by weight, 0.001% by weight, 0.005% by weight, 0.01% by weight, 0.05% by weight, 0.1% by weight or more, based on the total weight of the primary hydrophilic solution , At least 0.2 wt%, at least 0.3 wt%, at least 0.4 wt%, at least 0.5 wt%, at least 0.6 wt%, at least 0.7 wt%, at least 0.8 wt%, at least 0.9 wt%, at least 1 wt%, at least 2 wt% , 3% by weight, 4% by weight, 5% by weight, 10% by weight, 20% by weight, 30% by weight, 40% by weight, 50% by weight, 60% by weight, 70% by weight, 80 wt% or more or 90 wt% or more, and 99 wt% or less, 90 wt% or less, 80 wt% or less, 70 wt% or less, 60 wt% or less, 50 wt% or less, 40 wt% or less, 30 wt% % Or less, 20% or less, 10% or less, 9% or less, 8% or less, 7% or less, 6% or less, 5% or less, 4% or less, 3% or less , 2 wt% or less, or 1 wt% or less, and more specifically, 0.1 to 3 wt%, but is not limited thereto.

Micelle of the present invention can be prepared by mixing the dispersion and the primary hydrophilic solution. The micelle is a colloidal dispersion state, and when the polarity of the solvent is large, the micelle is formed with the polar polar group facing outward, and when the polarity of the solvent is small, the micelle is formed with the hydrophobic group facing outward. The micelle of the present invention may be an inverted micelle. In addition, the dispersion and the primary hydrophilic solution may be mixed in a volume ratio of 1: 0.1 to 10. Specifically, the volume ratio of the dispersion and the primary hydrophilic solution is 1 to 0.1 or more, 0.2 or more, 0.3 or more, 0.4 or more, 0.5 or more, 0.6 or more, 0.7 or more, 0.8 or more, 0.9 or more, 1 or more, 2 or more, 3 Or more, 4 or more, or 5 or more, 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, 2 or less, 1 or less, 0.9 or less, 0.8 or less, 0.7 or less, 0.6 Less than or equal to 0.5, less than or equal to 0.4, less than or equal to 0.3, less than or equal to 0.2, or less than 0.1, more specifically 1: 0.5, but is not limited thereto.

The micelle of the present invention may be a mixture of the dispersion and the primary hydrophilic solution using a high pressure emulsifier. The high pressure emulsifier may be used to obtain a uniform mixture. The high pressure emulsifier may be a microfluidizer or a two-stage homogenizer, but is not limited thereto.

Secondary hydrophilic active substances of the present invention include branched amino acids, hydrophilic drugs and the like. The branched amino acid is an amino acid having a branched alkyl group in the side chain, and specifically, may be valine, leucine, or isoleucine, but is not limited thereto. The hydrophilic drug may be, but is not limited to, a cell membrane dye, a photosensitizer, an anticancer agent, or an antibiotic.

Specifically, the secondary hydrophilic solvent of the present invention may be water, but the secondary hydrophilic solvent is not limited thereto.

The secondary hydrophilic solution of the present invention is to dissolve the secondary hydrophilic active material in the secondary hydrophilic solvent, the secondary hydrophilic active material may be included in 0.0001 to 99% by weight relative to the total weight of the secondary hydrophilic solution. Specifically, the secondary hydrophilic active material is at least 0.0001% by weight, at least 0.0005% by weight, at least 0.001% by weight, at least 0.005% by weight, at least 0.01% by weight, at least 0.05% by weight, and at least 0.1% by weight, based on the total weight of the secondary hydrophilic solution. , At least 0.2 wt%, at least 0.3 wt%, at least 0.4 wt%, at least 0.5 wt%, at least 0.6 wt%, at least 0.7 wt%, at least 0.8 wt%, at least 0.9 wt%, at least 1 wt%, at least 2 wt% , 3% by weight, 4% by weight, 5% by weight, 10% by weight, 20% by weight, 30% by weight, 40% by weight, 50% by weight, 60% by weight, 70% by weight, 80 wt% or more or 90 wt% or more, and 99 wt% or less, 90 wt% or less, 80 wt% or less, 70 wt% or less, 60 wt% or less, 50 wt% or less, 40 wt% or less, 30 wt% % Or less, 20% or less, 10% or less, 9% or less, 8% or less, 7% or less, 6% or less, 5% or less, 4% or less, 3% or less , 2 wt% or less, or 1 wt% or less, and more specifically, 0.1 to 3 wt%, but is not limited thereto.

The content of the primary hydrophilic active material, the primary hydrophilic solvent and the primary hydrophilic active material may be the same as or different from those of the secondary hydrophilic active material, the secondary hydrophilic solvent and the secondary hydrophilic active material.

The multiple emulsion of the present invention can be prepared by mixing the micelles and the secondary hydrophilic solution. The multiphase emulsion refers to an emulsion in which an oil phase is dispersed in an aqueous phase and contains a fine oil phase again in the oil phase, or an emulsion in which the oil phase is dispersed and a fine oil phase is contained in the aqueous phase again. It is shown when inverting the phase by adding another emulsifier from the place, or when making an emulsion using two kinds of emulsifiers from the beginning. The multiphase emulsion of the present invention is an emulsion of W / O / W type (Water / Oil / Water type). Can be. In addition, the micelles and the secondary hydrophilic solution may be mixed in a volume ratio of 1: 0.1 to 10. Specifically, the volume ratio of the micelle and the secondary hydrophilic solution is 1 to 0.1 or more, 0.2 or more, 0.3 or more, 0.4 or more, 0.5 or more, 0.6 or more, 0.7 or more, 0.8 or more, 0.9 or more, 1 or more, 2 or more, 3 Or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, or 9 or more, 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, 2 or less, or 1 It may be less than or equal to 1: More specifically, but is not limited thereto.

The multiphase emulsion of the present invention may be a mixture of the micelle and the secondary hydrophilic solution using a high pressure emulsion. The high pressure emulsifier may be used to obtain a uniform mixture. The high pressure emulsifier may be a microfluidizer or a two-stage homogenizer, but is not limited thereto.

The liposome solution of the present invention may be one in which the alcohol is removed from the multiphase emulsion, and the alcohol removal may be using a concentrator. According to one embodiment of the present invention, when spray drying is used to prepare liposome powder, the material to be dried should be present at a specific concentration (eg, 10% or more). The liposome solution of high concentration may be prepared by replacing the solvent volatilization process with a concentration process, and the liposome solution may have a liposome content of 10% or more.

Particle size or average particle size of liposomes in liposome solution of the present invention may be specifically 90 to 140 nm, more specifically 90 nm or more, 92 nm or more, 94 nm or more, 96 nm or more, 98 nm or more, 100 nm 102 nm or more, 104 nm or more, 106 nm or more, 108 nm or more, 110 nm or more, 111 nm or more, 112 nm or more, 113 nm or more, 114 nm or more, 115 nm or more, 116 nm or more, 117 nm or more, 118 nm or more, 119 nm or more, or 120 nm or more, 140 nm or less, 138 nm or less, 136 nm or less, 134 nm or less, 132 nm or less, 130 nm or less, 129 nm or less, 128 nm or less, 127 nm or less, 126 nm or less, 125 nm or less, 124 nm or less, 123 nm or less, 122 nm or less, 121 nm or less, 120 nm or less, 119 nm or less, 118 nm or less, 117 nm or less, 116 nm or less, 115 nm or less, 114 nm 113 nm or less, 112 nm or less, 111 nm or less, 110 nm or less, 109 nm or less, 108 nm or less, 107 nm or less, 106 nm or less, 105 nm or less, 104 nm or less, 103 nm or less, 102 nm or less , It can be not more than 101 nm or less, or 100 nm. According to one embodiment of the present invention, the particle size of the liposome in the liposome solution was confirmed that the average size is 120 nm or less, the distribution is uniform PDI (polydiversity index) of 0.02 or less (Experimental Example 1 and Figure 2).

The liposome powder of the present invention may be prepared by spray drying the liposome solution, wherein the spray drying is rapidly dried while spraying and dispersing the liquid material in hot wind to return the hot air powder product It may be a drying method obtained. The conventional liposome powdering process using freeze drying increases the production cost due to the use of cryoprotectants such as sugar or isopropyl alcohol, which is a lyophilization aid, to prevent the destruction of liposomes during freeze drying. It was very disadvantageous. Therefore, according to one embodiment of the present invention, the liposome powdering process is carried out using spray drying instead of freeze drying, which is more inexpensive, and can be directly applied industrially, and is easily applied to various industries such as food and cosmetics. Yes).

The particle size or average particle size of the liposome of the liposome powder of the present invention may be specifically 90 to 140 nm, more specifically 90 nm or more, 92 nm or more, 94 nm or more, 96 nm or more, 98 nm or more, 100 nm 102 nm or more, 104 nm or more, 106 nm or more, 108 nm or more, 110 nm or more, 111 nm or more, 112 nm or more, 113 nm or more, 114 nm or more, 115 nm or more, 116 nm or more, 117 nm or more, 118 nm or more, 119 nm or more, or 120 nm or more, 140 nm or less, 138 nm or less, 136 nm or less, 134 nm or less, 132 nm or less, 130 nm or less, 129 nm or less, 128 nm or less, 127 nm or less, 126 nm or less, 125 nm or less, 124 nm or less, 123 nm or less, 122 nm or less, 121 nm or less, 120 nm or less, 119 nm or less, 118 nm or less, 117 nm or less, 116 nm or less, 115 nm or less, 114 nm 113 nm or less, 112 nm or less, 111 nm or less, 110 nm or less, 109 nm or less, 108 nm or less, 107 nm or less, 106 nm or less, 105 nm or less, 104 nm or less, 103 nm or less, 102 nm or less , It can be not more than 101 nm or less, or 100 nm. According to one embodiment of the invention, the liposome powder has a spherical uniform form (Experimental Examples 2, 4 and 5), the particle size of the liposome in the liposome powder is similar to that of the liposome solution to spray drying Liposomes prepared through it was confirmed that the size and uniformity of the liposomes are well maintained without breaking (Experimental Example 1 and Figure 3).

The present invention provides a liposome prepared by the liposome preparation method. Liposomes, description of the liposome manufacturing method is as described above.

In the above aspects, the present invention provides a composition, food composition, cosmetic composition or pharmaceutical composition for delivery of the active material comprising the liposome.

Active substance delivery of the present invention is intended to maximize the activity of the active substance affecting metabolism in the human body and to minimize the side effects of the human body, specifically, the active substance may be a drug, when the active substance is a drug In addition, the present invention may include a drug delivery system (DDS) for the purpose of maximizing the therapeutic effect of the medicine and minimizing the side effects of the human body. Active substances administered to the body through various routes reach specific sites by moving to biological membranes and organs through transport routes such as blood and lymphatic system.

The present invention provides a food composition comprising the liposome. The composition according to the present invention may be included in food, and may have a powder, suspension, semi-solid formulation, tablet or capsule formulation, and when the liposome of the present invention is used as a food additive, it is added as it is separated from the synthesis or extract. It may be used in conjunction with other food or food ingredients, and may be appropriately used according to a conventional method. In addition, the mixed amount of the liposome may be used by appropriately adjusted according to the purpose of use (prevention, health or therapeutic treatment).

There is no particular limitation on the kind of food. Examples of the food to which the substance may be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, and drinks. Alcoholic beverages and vitamin complexes, and all foods in the conventional sense.

The food supplement additive of the present invention may use various flavors or natural carbohydrates. The above-mentioned natural carbohydrates are glucose, monosaccharides such as fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as tautin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like can be used.

In addition to the above, the composition according to the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, Alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the composition according to the present invention may contain a flesh for preparing natural fruit juice, fruit juice beverage and vegetable beverage. These components can be used independently or in combination.

The present invention also provides a cosmetic composition comprising the liposome. It can be prepared in any of the known formulations and formulated in the form of lotion, nourishing lotion, nourishing cream, massage cream, essence, pack, paste, gel, cream, lotion, powder, soap, oil, foundation, wax or spray. Can be.

In the cosmetic composition of each of the above formulations, other components in addition to the liposomes can be arbitrarily selected and blended according to the formulation or purpose of use of other cosmetics. In addition, the compositions of each formulation may contain a variety of bases and additives necessary for the formulation of the formulation and are suitable, and nonionic surfactants, silicone polymers, extender pigments, fragrances, preservatives, Fungicides, oxidation stabilizers, organic solvents, ionic or nonionic thickeners, softening agents, antioxidants, free radical destroying agents, opacifying agents, stabilizers, emollients, silicones, α-hydroxy acids, defoamers, humectants, Known compounds such as vitamins, insect repellents, fragrances, preservatives, surfactants, anti-inflammatory agents, substance P antagonists, fillers, polymers, propellants, basicizing or acidifying agents, or coloring agents.

The present invention also provides a pharmaceutical composition comprising the liposome. The liposome may further include an active substance effective for a specific disease as an active ingredient, and in addition to the active ingredient, additional ingredients, pharmaceutically acceptable or nutritionally acceptable carriers, depending on the formulation, method of use, and purpose of use. It may further comprise excipients, diluents or subcomponents.

Specifically, the pharmaceutical composition comprising the liposome may further contain a nutrient, vitamins, electrolytes, flavors, colorants, neutralizers, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like may further be contained.

As the carrier, excipient and diluent, all conventional ones can be used, and examples thereof include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, Calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, calcium carbonate, dextrin, propylene glycol, liquid paraffin And physiological saline may be one or more selected from the group consisting of, but is not limited thereto. The components can be added independently or in combination with liposomes.

The pharmaceutical composition comprising the liposome according to the present invention may be used alone or in combination with surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers.

Liposomes according to the present invention can be administered orally or parenterally (eg, intravenously, subcutaneously, intraperitoneally or topically) according to the desired method, and the dosage is determined by the weight, age, sex, health of the patient. The range varies depending on the condition, diet, time of administration, method of administration, rate of excretion and the severity of the disease. The daily dosage of the liposome of the present invention is 0.01 to 10000 mg / kg, preferably 1 to 20 mg / kg, preferably administered once to three times a day.

In addition, the pharmaceutical composition comprising the liposome of the present invention provides a medicament for the prevention or treatment of specific diseases, the medicament is a granule, powder, syrup, liquid, suspension, tablets, capsules, troches, pills, injections It may be, but is not limited to, a drug formulated as a transdermal absorbent, lotion, ointment, patch, cataplasm, paste or suppository.

When the formulation of the composition of the present invention is a paste, cream or gel, vegetable oils, waxes, paraffins, starches, trachants, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide may be used as carrier components. have.

When the formulation of the composition of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, in particular, in the case of a spray, additionally chlorofluorohydrocarbons. Propellant, such as propane / butane or dimethyl ether.

When the formulation of the composition of the present invention is a solution or emulsion, a solvent, solubilizing agent or emulsifying agent is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol , Fatty acid esters of 1,3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.

When the dosage form of the composition of the present invention is a suspension, a carrier diluent such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Microcrystalline cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used.

Hereinafter, the present invention will be described through the following examples and experimental examples. The following Examples and Experimental Examples are provided only for the purpose of illustration in order to facilitate understanding of the present invention, and the scope and scope of the present invention are not limited thereto.

[ Example Of the present invention Liposomes  Produce

A dispersion was prepared by dispersing 0.5% by weight of soybean-derived lecithin (ES food raw material) and 0.0625% by weight of β-sitosterol (Sigma) using agitation or a high pressure emulsified nano-device. It was. In addition, primary hydrophilic solutions were prepared by dissolving 2% by weight of branched amino acids valine, leucine and isoleucine (Sigma) as distilled water as primary hydrophilic active substances. Then, the dispersion and the primary hydrophilic solution were mixed for 1 minute in a homogenizer (homogenizer) at a volume ratio of 2: 1 to prepare an oil-in-water emulsion, and the oil-in-water emulsion was prepared at 1000 bar, 10 Inverted micelles were made using a microfluidizer in a pass condition or a two-stage homogenizer in a 400 bar, 10 pass condition.

Thereafter, 1% by weight of branched amino acids valine, leucine and isoleucine (sigma) as distilled water were added to distilled water as a secondary hydrophilic active material, and stirred for 5 minutes to dissolve to prepare a secondary hydrophilic solution. Then, the solution containing the inverted micelle and the secondary hydrophilic solution were mixed in a homogenizer for 1 minute at a volume ratio of 1: 2 to prepare a W / O / W type polyphase emulsion (W / O / W emulsion). The W / O / W emulsion was prepared and homogenized using a 500 bar, 5 pass microfluidizer or 200 bar, 5 pass condition two-stage homogenizer. Thereafter, liquor in the W / O / W emulsion was removed using a concentrator for 4 hours to form a liposome solution. In addition, the liposome solution was spray dried (spray drying) to prepare a liposome powder, the liposome manufacturing process of the present invention is shown in FIG.

[ Experimental Example  One] Liposome  Check Particle Size and Uniformity

In order to measure the liposome particle size of each of the liposome solution and the liposome powder prepared in the above example, a dynamic light scattering method using zetasizer nano ZS (Malvern instruments Ltd, 1XZ, UK) DLS) was performed in the following manner.

Specifically, 1.0 mL of the liposome solution prepared in Example was placed in a disposable plastic cuvette, and the DLS was repeatedly performed three times under the following conditions, and the results are shown in FIGS. 2 and 3.

Measurement conditions: refractive index (1.330), speed (0.8872 cP), equilibration time (1 min), temperature (25 ° C), measurement angle (173 ° backscattering)

As shown in Figure 2, it was found that the particle size of the liposome solution prepared by using the alcohol liposomes of the nanoparticles of uniform size with an average size of 120 nm or less, PDI (polydiversity index) = 0.02 or less. In addition, as shown in FIG. 3, the liposome powder also has a particle size and uniformity similar to that of the liposome solution, whereby the liposome prepared by spray drying shows that the size and uniformity of the liposome is well maintained without being destroyed. Could.

[ Experimental Example  2] Liposome  Morphological analysis

The shape of the powdered liposomes prepared in the above example was analyzed using a Transmission Electron Microscope (TEM). In the course of analysis, uranyl acetate was used as a negative staining reagent of liposomes.

Specifically, 10 μL of the 10% liposome powder solution in which the powdered liposomes prepared in the above example was dissolved was formed into a formvar grid (200 mesh) coated with silicon monoxide. After 1 minute, a 2% uranyl acetate solution was placed on the grid. After 1 minute, the grid was washed with distilled water, the grid washed at room temperature and dried, and then observed the form of liposomes by TEM (120 keV, JEOL Ltd., Tokyo, Japan), the results are shown in Figures 4 and 5 Shown in

As shown in Figures 4 and 5, the liposomes prepared using the alcohol has a spherical uniform shape, it can be seen that even after spray drying the form is maintained and not destroyed.

Formulation examples of the composition according to an embodiment of the present invention will be described below, but can be applied to other various formulations, which are intended to explain in detail only, not intended to limit the present invention.

Formulation Example 1 Preparation of Nutrients

According to the composition shown in Table 1 it was prepared nutrient cosmetics in a conventional manner.

Raw material name Content (% by weight) glycerin 3.0 Butylene glycol 3.0 Propylene glycol 3.0 Carboxy Vinyl Polymer 0.1 Example Powdered Liposomes 1.0 Beeswax 4.0 Polysorbate 60 1.5 Caprylic / Capric Reglycerides 5.0 Squalane 5.0 Sorbitassquioleate 1.5 Cetearyl Alcohol 1.0 Triethanolamine 0.2 Preservatives, spices Quantity Purified water Remaining amount system 100

Formulation Example 2 Nutritional Lotion

According to the composition shown in Table 2 it can be prepared by the conventional method lotion.

Raw material name Content (% by weight) Purified water Remaining amount glycerin 3.0 Butylene glycol 3.0 Liquid paraffin 5.0 Beta Glucan 7.0 Carbomer 0.1 Example Powdered Liposomes 1.0 Caprylic / Capric Triglycerides 3.0 Squalane 5.0 Cetearyl Glucoside 1.5 Sorbitan stearate 0.4 Polysorbate 60 1.5 antiseptic Quantity incense Quantity Pigment Quantity Triethanolamine 0.1 system 100

Formulation Example 3 Nutrition Cream

Nutritional creams may be prepared by conventional methods according to the composition described in Table 3.

Raw material name Content (% by weight) glycerin 3.5 Butylene glycol 3.0 Liquid paraffin 7.0 Beta Glucan 7.0 Carbomer 0.1 Example Powdered Liposomes 1.0 Caprylic / Capric Triglycerides 3.0 Squalane 5.0 Cetearyl Glucoside 1.5 Sorbitan stearate 0.4 Polysorbate 60 1.2 Triethanolamine 0.1 Preservatives, spices Quantity Purified water Remaining amount system 100

[Formulation Example 4] Hair Pack

The pack can be prepared by conventional methods according to the compositions described in Table 4 below.

Raw material name Content (% by weight) glycerin 4.0 Polyvinyl alcohol 15.0 Hyaluronic acid extract 5.0 Beta Glucan 7.0 Allantoin 0.1 Example Powdered Liposomes 1.0 Nonyl Phenyl Ether 0.4 Polysorbate 60 1.2 Ethanol preservative Quantity Preservatives, spices Quantity Purified water Remaining amount system 100

Formulation Example 5 Ointment

Ointments can be prepared by conventional methods according to the compositions described in Table 5 below.

Raw material name Content (% by weight) Example Powdered Liposomes 2.0 Beta-1,3-glucan 10.0 Beeswax 10.0 Polysorbate 5.0 Fiji 60 Cured Castor Oil 2.0 Sorbitan sesquioleate 0.5 vaseline 5.0 Liquid paraffin 10.0 Squalane 5.0 Shea Butter 3.0 Caprylic / Capric Triglyceride 5.0 glycerin 10.0 Propylene glycol 10.2 Triethanolamine 0.2 Preservative, coloring, flavoring Quantity Purified water Remaining amount system 100

Formulation Example 6 Preparation of Pharmaceutical Agent (Patch) for Local Administration

In accordance with the composition shown in Table 6 below, the pharmaceutical preparation (patch) for topical administration was prepared in a conventional manner.

Raw material name Content (% by weight) Example Powdered Liposomes 2.0 Beta-1,3-glucan 3.0 Diethylamine 0.7 Sodium sulfite 0.1 Polyoxyethylene lauryl ether (E.O = 9) 1.0 Polyhydroxyethylenecetylstearyl ether (Cetomacrogol 1000) 1.0 Viscous Paraffin Oil 2.5 Caprylic Acid Ester / Capric Acid Ester (Cetiol LC) 2.5 Polyethylene Glycol 400 3.0 Polyacrylic Acid (Carbopol 934P) 1.0 Purified water Remaining amount system 100

Formulation Example 7 Preparation of Powder

Example 2 Powdered Liposomes

1g lactose

After mixing the above components and filled in airtight cloth to prepare a powder.

Formulation Example 8 Preparation of Tablet

Example 100 mg of powdered liposomes

Corn Starch 100mg

Lactose 100mg

2 mg magnesium stearate

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

Formulation Example 9 Preparation of Capsule

Example 100 mg of powdered liposomes

Corn Starch 100mg

Lactose 100mg

2 mg magnesium stearate

After mixing the above components, the capsule was prepared by filling in gelatin capsules according to the conventional method for producing a capsule.

Formulation Example 10 Preparation of Ring

Example 1 Powdered Liposome 1g

Lactose 1.5g

1 g of glycerin

Xylitol 0.5g

After mixing the above components, it was prepared to be 4g per ring according to a conventional method.

Formulation Example 11 Preparation of Granules

Example 150g of Powdered Liposomes

Soybean Extract 50mg

Glucose 200mg

Starch 600mg

After mixing the above components, 100mg of 30% ethanol was added and dried at 60 degrees Celsius to form granules, and then filled into fabric.

Formulation Example 12 Drinks

50 mg of powdered liposomes of Example 1

10 g of glucose

0.6 g citric acid

25g of liquid oligosaccharides

After mixing the above components, add 300 ml of purified water and fill each bottle with 200 ml. After filling the bottle sterilized for 4-5 seconds at 130 ℃ to prepare a beverage.

Formulation Example 13 Caramel Formulations

Example 50 mg of powdered liposomes

Corn syrup 1.8g

0.5 g skim milk

0.5 g soy lecithin

0.6 g butter

0.4g of vegetable hardened oil

1.4 g of sugar

Margarine 0.58g

20 mg of salt

Caramel molding can be performed by mixing the above components.

Claims (21)

Preparing a micelle by mixing a dispersion obtained by dispersing phospholipid in alcohol and a primary hydrophilic solution in which a primary hydrophilic active material is dissolved in a primary hydrophilic solvent by a high pressure emulsifying apparatus;
Preparing a multiple emulsion by mixing a secondary hydrophilic solution in which a secondary hydrophilic active material is dissolved in a secondary hydrophilic solvent and the prepared micelle with a high pressure emulsion apparatus;
Removing liquor using a concentrator in the prepared multiphase emulsion and preparing a liposome solution having a liposome concentration of 10% or more; And
Spray drying the liposome solution to prepare a liposome powder,
The high pressure emulsifying apparatus is a microfluidizer or a two-stage homogenizer, liposome manufacturing method. The method of claim 1, wherein the phospholipid is lecithin. The method of claim 1, wherein the alcohol is 90-100% alcohol. The method of claim 1, wherein the content of the phospholipid is 0.1 to 1% by weight based on the total weight of the dispersion. The method of claim 1, wherein the primary hydrophilic solvent is water. The method of claim 1, wherein the content of the primary hydrophilic active material is 0.0001 to 99% by weight based on the total weight of the primary hydrophilic solution. The method of claim 1, wherein the volume ratio of the dispersion and the primary hydrophilic solution is 1: 0.1 to 10. delete delete The method of claim 1, wherein the secondary hydrophilic solvent is water. The method of claim 1, wherein the content of the secondary hydrophilic active material is 0.0001 to 99% by weight based on the total weight of the secondary hydrophilic solution. The method of claim 1, wherein the volume ratio of the micelles and the secondary hydrophilic solution is 1: 0.1 to 10, liposome production method. The method of claim 1, wherein the micelle is an inverted micelle. The method of claim 1, wherein the multiphase emulsion is W / O / W type (Water / Oil / Water type). The method of claim 1, wherein the particle size of the liposome in the liposome solution is 90 to 140 nm. The method of claim 1, wherein the liposome powder has a particle size of 90 to 140 nm. A liposome prepared by the method of any one of claims 1 to 7, 10 to 16. 18. An active substance delivery composition comprising the liposome of claim 17. A food composition comprising the liposome of claim 17. A cosmetic composition comprising the liposome of claim 17. A pharmaceutical composition comprising the liposome of claim 17.

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