KR20220131805A - Lipid nanoparticle composition prepared with mastic gum or gamma oryzanol - Google Patents

Abstract

The present invention relates to lipid nanoparticles having a high encapsulation rate of physiologically active substances and high absorbability by using mastic gum or gammaoryzanol, or a composition thereof, and a method for manufacturing the lipid nanoparticles having reduced toxicity by not using harmful organic solvents or the composition thereof.

Description

Lipid nanoparticle composition prepared with mastic gum or gamma oryzanol {Lipid nanoparticle composition prepared with mastic gum or gamma oryzanol}

The present invention relates to a method for improving dispersion stability and absorption by improving the composition of lipid nanoparticles.

Lipid nanoparticles refer to lipids made of small particles less than 1 micron in size.

However, since there are numerous types and forms of lipids, there are many types and forms of lipid nanoparticles, but there is no standard for systematically classifying lipid nanoparticles. In general, if the lipid used for lipid nanoparticles is a solid at room temperature, it is called a solid lipid nanoparticle (SLN). , NLC). Compared to SLN, NLC is an advanced technology that can prevent recrystallization by inhibiting the movement of encapsulated materials in lipid nanoparticles. Therefore, recently, NLC rather than SLN has been widely used, and it is common to call the two types of lipid nanoparticles as lipid nanoparticles (NLP) without needing to distinguish them.

Lipid nanoparticles are widely used materials in the drug delivery system (DDS) area, and while maximizing the desired effect, such as when you want to selectively deliver a drug to a target site, or maintain an effective blood concentration for a long time. It is a useful technique for minimizing side effects. Recently, various manufacturing methods are being developed as the number of industrial fields to be applied as new materials using lipid nanoparticles increases.

Representative manufacturing methods include high-pressure homogenization, emulsification-sonification, microemulsion, solvent emulsification-evaporation, solvent diffusion, and solvent injection. (solvent injection), multiple emulsion (multiple emulsion), high-pressure dispersion emulsification (microfluidizer emulsification), etc. are well known, and also a method of mixing these methods and other various manufacturing methods.

When lipid nanoparticles are to be applied as part of a drug delivery system, since the target site to be applied is a part of the human body, the lipid nanoparticles should be well dispersed in water and, after dispersion, should have excellent dispersion stability. In this regard, lipid nanoparticles as drug delivery systems still have many problems to be improved.

On the other hand, when phospholipids are used among lipid nanoparticles, they are also called phospholipid nanoparticles. As mentioned above, lipid nanoparticles used in drug delivery systems often use phospholipids because their goal is to apply them to the human body. This is in consideration of the fact that there are many phospholipids among the lipids constituting the human body, and in particular, the membranes of cells constituting the human body are phospholipids.

A representative example of lipid nanoparticles is a liposome. Therefore, the preparation method of lipid nanoparticles is very similar to the preparation method of liposomes. So, the manufacturing method of lipid nanoparticles is thin film method, ultrasound method, reverse-phase evaporation method, ether vaporization method, freeze-drying extraction method ( freeze-thaw extrusion method, dehydration-rehydration method, microfluidizer emulsification and the like are used.

No matter what method is used for lipid nanoparticles, a homogenization process of dispersing them in water to form nanoparticles is essential in the manufacturing process. However, in the case of mechanical homogenization, oxygen in the air is introduced during the homogenization process to oxidize the lipids, thereby generating lipid peroxides, so there is a problem in that the lipid layer is degraded. Lipid denaturation adversely affects the dispersion stability of lipid nanoparticles. In addition, if an ultrasonic process is used instead of mechanical homogenization, titanium is separated from the probe tip and heavy metals are mixed into the finished product. Even a small amount of heavy metals mixed into the lipid nanoparticle dispersion can cause fatal toxicity to the human body.

On the other hand, phospholipids are basically used for lipid nanoparticles, and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC),1,2-dioleoyl-sn-glycero-3-phosphoethanolamine ( DOPE), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) and dipalmitoyl-phosphatidylglycerol (DPPG) are representative phospholipids. Also, in the composition of lipid nanoparticles, cholesterol is used together as in the case of liposomes, and a third lipid other than phospholipids is additionally used.

The use of cholesterol in the composition of lipid nanoparticles and liposomes is an application of the fact that the cell membranes of all living things are composed of phospholipids and cholesterol. In fact, cholesterol-free lipid membranes are very unstable and are easily destroyed, whereas lipid membranes made by adding an appropriate amount of cholesterol secure fluidity and rigidity to form a stable structure, helping to stabilize the dispersion of nanoparticles.

In the case of making lipid nanoparticles for use in drug delivery systems, phospholipids and cholesterol are the basic compositions by mimicking the cell membrane of an organism, and additional lipids are added to achieve desired purposes such as stability and absorption. .

As described above, cholesterol is used in lipid nanoparticles to be applied as a drug delivery system. However, the process of artificially manufacturing lipid nanoparticles containing cholesterol is completely different from the situation that occurs in living organisms. Cholesterol synthesis in living organisms starts from acetyl-CoA, a water-soluble substance, and goes through a chain of enzymatic reactions to make it at the site where cholesterol is needed. Therefore, the fat solubility of cholesterol is not an issue. In contrast, in the case of artificially making lipid nanoparticles, a process of dissolving cholesterol to a molecular level is required first.

Cholesterol is not soluble in water or hydrophilic solvents, but only soluble in harmful organic solvents. Therefore, there is a problem in that the use of harmful organic solvents cannot be avoided in order to prepare lipid nanoparticles containing cholesterol. In order to develop a drug delivery system containing cholesterol as a formulation, harmful organic solvents must be removed before administration to the human body. A method for removing harmful organic solvents generally uses a vacuum rotary evaporator or a dialysis membrane, and although this process is expensive, it is a bigger problem that the organic solvent pollutes the environment.

When lipid nanoparticles to be developed as a drug delivery system must pass through the cell membrane as in the case of a Covid-19 vaccine, a separate synthetic lipid is added as a component to increase cell membrane permeability. However, synthetic lipids have a problem with low industrial performance due to a complicated manufacturing process.

U.S. Avanti company supplies synthetic lipids, such as DOTAP (N-[1-(2,3-Dioleoyloxy)propyl]-N,N,N-trimethylammonium chloride), DOTMA (1,2-di-O-octadecenyl-3 -trimethylammonium propane (chloride salt)) is a representative product. However, this synthetic lipid has a complicated manufacturing process, so it is mainly used only for research purposes, and it is difficult to use industrially.

Therefore, companies that have succeeded in commercializing using lipid nanoparticles are using synthetic lipids supplied by a separate company.

In the case of lipid nanoparticles developed by Alnylam in the United States as a treatment for polyneuropathy, (6Z,9Z,28Z,31Z)-heptatriacont-6,9,28,31-tetraene-19-yl 4- (dimethylamino)butanoate is used.

In the case of lipid nanoparticles developed as a Covid-19 vaccine by Pfeizer in the United States and BioNTech in Germany, synthetic lipids ((4-Hydroxybutyl) azanediyl) bis (hexane-6,1-diyl) bis 2-hexyldecanoate and phospholipids and cholesterol 2-[(polyethylene glycol)-2000]-N,N-ditetradecylacetamide is additionally used.

In the case of lipid nanoparticles developed by Moderna and the Institute of Allergy and Infectious Diseases as a Covid-19 vaccine, heptadecan-9-yl 8-((2-hydroxyethyl) (6-oxo-6-(undecyloxy) hexyl) together with phospholipids and cholesterol Synthetic lipids such as amino) octanoate and polyethylene glycol 2000 dimyristoyl glycerol are additionally used.

In summary, in order to artificially make lipid nanoparticles for application as drug delivery systems, cholesterol must be dissolved at the molecular level. At this time, the use of hazardous organic solvents is unavoidable, and the used hazardous organic solvents must be removed, causing problems such as cost and environmental pollution in this process. In addition, synthetic lipids are often used additionally to enhance the functionality of the drug delivery system, but there is also a problem with low industriality.

The present invention has been proposed to solve the above problems, and it is a cholesterol substitute material that can form stable lipid nanoparticles by being well mixed with other components of lipid nanoparticles at the molecular level without using harmful organic solvents to the human body. It aims to provide lipid nanoparticles or a composition thereof with improved dispersion stability and absorption by using the searched alternative material.

To achieve this object, the present inventors search for natural substances that can be used instead of cholesterol as a constituent material of lipid nanoparticles, and devise a method for manufacturing lipid nanoparticles without using an organic solvent harmful to the human body. wanted to As a result of repeated research, the inventors have found that when mastic or gamma oryzanol is included in lipid nanoparticles, the physiologically active substances can be loaded into the lipid nanoparticles by encapsulating them in a high content regardless of whether they are fat-soluble or water-soluble, without the presence of cholesterol. Confirmed. In addition, it was confirmed that there is no need for the use of harmful organic solvents required to dissolve cholesterol due to not using cholesterol, and toxicity is reduced, and there is no need for an additional process for removing the organic solvent. In addition, mastic gum is a liquid surfactant such as PEG, glycerol, propylene glycol, dipropylene glycol, sorbitan-fatty acid ester, sorbitan-PEG-fatty acid ester, castor oil-PEG derivative, etc. As a result of confirming that it cannot be dissolved at the molecular level, and seeking a new method beyond the conventional solubilization method, a method of mixing mastic gum or gamma oryzanol with phospholipids and oils and fats and then heating to melt it was introduced to avoid the use of harmful organic solvents. The present invention has been completed by devising a new method that can be mixed with phospholipids and oils without any need.

Accordingly, according to one aspect of the present invention, lipid nanoparticles including mastic or gamma oryzanol in lipids such as phospholipids or a composition including the lipid nanoparticles may be provided.

According to one aspect of the present invention, a physiologically active material delivery system in which a physiologically active material is introduced into the lipid nanoparticles or a composition thereof may be provided. The physiologically active material is a fat-soluble or water-soluble material, and may be a nutritional component, a drug, or the like, but is not limited thereto.

According to one aspect of the present invention, there may be provided a method for preparing lipid nanoparticles or a composition thereof, comprising mixing mastic or gamma oryzanol with lipids and fats and melting and mixing them.

In the method for producing lipid nanoparticles and a composition thereof according to the present invention, lipid nanoparticles are prepared without using harmful organic solvents and synthetic lipids harmful to the human body by replacing cholesterol essential in the process with mastic gum or gamma oryzanol can do. Furthermore, the lipid nanoparticle composition prepared by this preparation method has improved dispersion stability and absorption, and these lipid nanoparticles are very useful as a carrier that can excellently deliver various physiologically active substances, regardless of whether they are fat-soluble or water-soluble. .

1 is a manufacturing process diagram of mastic gum or gamma oryzanol-containing lipid nanoparticles loaded with a physiologically active substance and a composition thereof.
2 is a particle size distribution diagram of a dispersion of Composition 4 prepared using mastic gum or gamma oryzanol.
3 is an electron transmission micrograph of a composition 4 dispersion prepared from mastic gum.
4 is a view confirming the dispersion stability of lipid nanoparticles prepared using mastic gum or gamma oryzanol.
5 is a diagram comparing the properties of the receptor compartment after the percutaneous permeation test of Composition 1 and Composition 4 dispersions prepared using mastic gum or gamma oryzanol.
6 is a graph showing the results of the transdermal permeation test of the lipid nanoparticle dispersion containing dutasteride prepared using mastic gum or gamma oryzanol.
7 is a graph showing the results of the transdermal permeation test of finasteride-containing lipid nanoparticle dispersions prepared using mastic gum or gamma oryzanol.
8 is a graph showing the results of a transdermal permeation test of ketoprofen-containing lipid nanoparticle dispersions prepared using mastic gum or gamma oryzanol.
9 is a graph showing the results of a transdermal permeation test of lidocaine-containing lipid nanoparticle dispersions prepared using mastic gum or gamma oryzanol.
10 is a graph showing the results of the transdermal permeation test of the minoxidil-containing lipid nanoparticle dispersion prepared using mastic gum or gamma oryzanol.
11 is a graph showing the results of the transdermal permeation test of docetaxel-containing lipid nanoparticle dispersion prepared using mastic gum or gamma oryzanol.
12 is a graph showing the results of the transdermal permeation test of the tamsulosin-containing lipid nanoparticle dispersion prepared using mastic gum or gamma oryzanol.
13 is a graph showing the results of the transdermal permeation test of the KTTKS-palmitate-containing lipid nanoparticle dispersion prepared using mastic gum or gamma oryzanol.
14 is a graph showing the results of a transdermal permeation test of a dispersion of curcumin-containing lipid nanoparticles prepared using mastic gum or gamma oryzanol.
15 is a graph showing the results of a transdermal permeation test of a dispersion of resveratrol-containing lipid nanoparticles prepared using mastic gum or gamma oryzanol.
16 is a graph showing the results of the transdermal permeation test of the quercetin-containing lipid nanoparticle dispersion prepared using mastic gum or gamma oryzanol.
17 is a graph showing the results of a transdermal permeation test of a dispersion of lutein-containing phospholipid nanoparticles prepared using mastic gum or gamma oryzanol.
18 is a graph showing the results of a transdermal permeation test of a risperidone-containing lipid nanoparticle dispersion prepared using mastic gum or gamma oryzanol.
19 is a graph showing the results of a transdermal permeation test of a brinzolamide-containing lipid nanoparticle dispersion prepared using mastic gum or gamma oryzanol.
20 is a graph showing the results of a transdermal permeation test of a lipid nanoparticle dispersion containing only clophage prepared using mastic gum or gamma oryzanol.
21 is a graph showing the results of a transdermal permeation test of etoposide-containing lipid nanoparticle dispersions prepared using mastic gum or gamma oryzanol.
22 is a view showing the results of a transdermal permeation test of ascorbic acid-containing lipid nanoparticles prepared using mastic gum or gamma oryzanol.
23 is a diagram showing the results of a transdermal permeation test of lipid nanoparticles containing epidermal growth factor prepared using mastic gum or gamma oryzanol.
24 is a graph showing the results of a transdermal permeation test of glutathione-containing lipid nanoparticles prepared using mastic gum or gamma oryzanol.
25 is a graph showing the results of a transdermal permeation test of risedronate sodium-containing lipid nanoparticles prepared using mastic gum or gamma oryzanol.
26 is a graph showing the results of the transdermal permeation test of irinotecan-containing lipid nanoparticles prepared using mastic gum or gamma oryzanol.
27 is a graph showing the absorption test results of oral administration of glutathione-containing lipid nanoparticles prepared using gamma oryzanol.

Hereinafter, the present invention will be described in detail.

The present invention provides lipid nanoparticles containing mastic or gamma oryzanol as a component for delivery of physiologically active ingredients together with lipids in lipid nanoparticles, or a composition comprising the lipid nanoparticles. Unlike conventionally known lipid nanoparticles, in the present invention, the drug delivery efficiency is significantly increased by including mastic or gamma oryzanol, and at the same time, the health hazards caused by cholesterol can be significantly reduced by not using cholesterol, and reduced pressure It has the advantage that the evaporation or dialysis process can be omitted.

In one embodiment, the lipid may be a phospholipid, and the phospholipid may be extracted from soybean or egg yolk, etc., and may be a phospholipid having a neutral charge or a positive charge. Examples of such phospholipids include, but are not limited to, dilauroyl phosphatidylethanolamine, dimyristoyl phosphatidylethanolamine, dipalmitoyl phosphatidylethanolamine, Distearoyl phosphatidylethanolamine, dioleoyl phosphatidylethanolamine, dilinoleoyl phosphatidylethanolamine, 1-palmitoyl-2-oleoyl phosphatidylethanolamine (1-palmitoylethanolamine) -oleoyl phosphatidylethanolamine), 1,2-diphytanoyl-3-sn-phosphatidylethanolamine (1,2-diphytanoyl-3-sn-phosphatidylethanolamine), 1,2-dioleoyl-sn-glycero-3-phospho Poethanolamine (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine, DOPE), dilauroyl phosphatidylcholine, dimyristoyl phosphatidylcholine, dipalmitoyl phosphatidylcholine, dipalmitoyl phosphatidylcholine Distearoyl phosphatidylcholine, dioleoyl phosphatidylcholine, dilinoleoyl phosphatidylcholine, 1-palmitoyl-2-oleoyl phosphatidylcholine (1-palmitoyl-2-oleoylphosphatidylcholine) 1,2-Dipalmitoyl-sn-glycero-3-phosphatidylcholine (1,2-dipalmitoyl-sn-glycero -3-phosphocholine, DPPC), 1,2-diphytanoyl-3-sn-phosphatidylcholine (1,2-diphytanoyl-3-sn-phosphatidylcholine), 1,2-dioleoyl-sn-glycero-3- Phosphatidylcholine (1,2-dioleoyl-sn-glycero-3-phosphatidylcholine, DOPC), 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (1,2-dimyristoyl-sn-glycero-3-phosphocholine) , DMPC), 1,2-distearoyl-sn-glycero-3-phosphatidylcholine (1,2-distearoyl-sn-glycero-3-phosphocholine, DSPC), dipalmitoyl-phosphatidylglycerol (dipalmitoyl-phosphatidylglycerol, DPPG) dilauroyl phosphatidic acid, dimyristoyl phosphatidic acid, dipalmitoyl phosphatidic acid, distearoyl phosphatidic acid acid), dioleoyl phosphatidic acid, dilinoleoyl phosphatidic acid, 1-palmitoyl-2-oleoyl phosphatidic acid ) and 1,2-diphytanoyl-3-sn-phosphatidic acid may be at least one selected from the group consisting of 1,2-diphytanoyl-3-sn-phosphatidic acid.

In one embodiment, the amount of mastic gum or gamma oryzanol added is 0.05% to 50% by weight, 0.07% to 47% by weight, 0.09% to 45% by weight or 0.1% to 40% by weight based on the weight of the lipid. %, but is not limited thereto.

In one aspect, the lipid nanoparticles and the composition according to the present invention may further include fats and oils. The mastic or gamma oryzanol contained in the lipid nanoparticles and the composition according to the present invention is characterized in that it is melted together with the oil and lipid.

The oil may include vegetable oil or animal oil, and examples of such vegetable oil may be at least one selected from the group consisting of palm oil, palm kernel oil, linseed oil, soybean oil, rice bran oil, corn oil, olive oil and peanut oil. , Examples of animal fats and oils may be at least one selected from the group consisting of beef tallow, brisket, horse oil, shark oil, light oil (whale oil), krill oil and butter oil. As a result of using these oils and fats, when preparing lipid nanoparticles and compositions thereof, mastic gum or gamma oryzanol melts and mixes well at the molecular level with lipids heated to 90° C. or higher and these oils and fats, so that nanoparticles are not needed without the need for substances such as cholesterol. is well formed.

In addition, the lipid nanoparticles or the composition of the present invention may further include a surfactant. Examples of the surfactant are not limited, but at least one liquid surfactant selected from the group consisting of PEG, glycerol, propylene glycol, dipropylene glycol, sorbitan-fatty acid ester, sorbitan-PEG-fatty acid ester and castor oil-PEG derivative. may be an active agent.

In addition, the lipid nanoparticles and the composition of the present invention may further include a pH adjusting agent, an isotonic agent, a preservative, an antioxidant, and the like.

In the present invention, the lipid nanoparticles and compositions thereof may have a formulation selected from the group consisting of liposomes, micelles, emulsions, and solid lipid nanoparticles.

On the other hand, the size of the lipid nanoparticles according to the present invention is not particularly limited. In one embodiment, the lipid nanoparticles may have a particle diameter of 10 to 500 nm. In general, it can be made into nanoparticles having a substantially uniform size in a particle diameter range of about 10 nm to 500 nm, 50 nm to 300 nm. For example, in the case of liposomes, an aqueous suspension of liposomes is passed through a series of polycarbonate membranes having a uniform membrane pore size selected in the range of 30 nm to 200 nm, typically 50 nm, 100 nm or 200 nm. By extrusion (extrusion), it is possible to prepare liposome nanoparticles having an equivalent size. The size of the membrane pores approximately corresponds to the average particle size of liposomes produced by extrusion through the membrane, and in particular, in order to homogenize the size of the prepared liposomes, several times, for example, 3 times or more through a membrane filter of the same membrane size. extrude.

In addition, the homogenization method can be usefully used to control the particle size of the liposome to a size less than a certain size, preferably less than 300 nm.

The lipid nanoparticles or a composition thereof according to the present invention may be used for drug delivery. Accordingly, the present invention provides a composition for drug delivery comprising the lipid nanoparticles.

In one embodiment, the lipid nanoparticles or a composition thereof may be for tissue-specific drug delivery.

The present invention also provides a lipid nanoparticle-bioactive substance complex in which a drug is introduced into the lipid nanoparticle obtained by adding mastic or gamma oryzanol to the lipid described above.

As described above, the lipid nanoparticles or a composition thereof according to the present invention can be used as a carrier of a physiologically active substance such as a drug or a nutrient component. Therefore, a desired physiologically active material is introduced into the lipid nanoparticles to form lipid nanoparticles loaded with a physiologically active material, and then the physiologically active material can be used for diagnosis, prevention or treatment of a target disease.

The type of physiologically active material that can be introduced into the lipid nanoparticles or the composition according to the present invention is not particularly limited. The physiologically active material may be introduced into the lipid nanoparticles according to the present invention through a hydrophobic bond. Alternatively, since there is a space in which the physiologically active material can be loaded or loaded in the interior of the nanoparticles, the physiologically active material in this internal space It is also possible to form a lipid nanoparticle-bioactive material complex by loading.

As used herein, the term "biologically active substance" is a substance that enhances or inhibits a biological function, and a substance that can maintain the balance when a substance that regulates the function of the human body is deficient or excessively secreted to show an abnormal condition means The physiologically active material may be any one or more selected from the group consisting of compounds, extracts, proteins, peptides, nucleic acids and antibodies.

In the present invention, the nucleic acid is ribonucleic acid (RNA), small interfering ribonucleic acid (small interfering RNA), antisense oligonucleotide, deoxyribonucleic acid (DNA), plasmid deoxyribonucleic acid, aptamer (aptamer) etc. Preferably, a small interfering ribonucleic acid consisting of 5 to 200 base pairs, an antisense nucleic acid, or a nucleic acid aptamer may be used.

On the other hand, the backbone, sugar or base of the nucleic acid may be chemically modified or the terminal thereof may be modified for the purpose of increasing blood stability or weakening an immune response. Specifically, a part of the phosphodiester bond of the nucleic acid is replaced with a phosphorothioate or boranophosphate bond, or a methyl group or a methoxyethyl group is located at the 2'-OH position of some ribose bases. , may include one or more modified nucleotides into which various functional groups such as fluorine are introduced.

In another embodiment, the physiologically active substance may be a compound. The kind of compound that can be introduced into the nanoparticles of the present invention is not particularly limited. The extract, protein, peptide, nucleotide or antibody exhibits physiological activity, and any material used for the prevention, treatment or improvement of diseases may be captured in the lipid nanoparticles of the present invention. The extract, protein peptide, nucleotide or antibody may be prepared by any method known in the art, and may be used in a required amount if necessary. In addition, any material exhibiting physiological activity may be captured in the lipid nanoparticles of the present invention.

As another example, the physiologically active material may be a fat-soluble material or a water-soluble material. In one embodiment, the fat-soluble drug is dutasteride, finasteride, ketoprofen, lidocaine, minoxidil, docetaxel, tamsulosin, KTTS-palmitate, resveratrol, quercetin, lutein, curcumin, risperidone, brinzolamide, clofa Jimin, etoposide, and the like, and the water-soluble drug may include, but is not limited to, ascorbic acid, epidermal growth factor (EGF), glutathione, risedronate, irinotecan, and the like.

Dutasteride and finasteride are substances used to treat benign prostatic hyperplasia by inhibiting 5-alpha reductase and preventing the conversion of testosterone to disiderotosterone. This substance is also used to treat alopecia.

Ketoprofen is a kind of nonsteroidal anti-inflammatory analgesic and is widely used to control pain caused by rheumatoid arthritis and various inflammations.

Lidocaine inactivates fast voltage-gated Na+ channels in the cell membrane and blocks signal transduction, so it is used in various diseases such as local anesthetics, pain regulators for various pain-inducing diseases such as herpes zoster, arrhythmias, and epilepsy. is a substance

Minoxidil is a substance used to treat high blood pressure that dilates blood vessels by opening potassium channels. Since it also expands blood vessels distributed in hair follicle cells of the scalp, it is also helpful in the treatment of alopecia.

Docetaxel is a kind of anticancer drug that inhibits the proliferation of cancer cells by interfering with the process of separation of microtubules, a mechanism of division and self-replication, during cell division. Docetaxel is a material with a relatively large molecular weight with a molecular weight of 807.9 g/mole.

Tamsulosin is a substance that selectively inhibits alpha-1A receptors distributed in the smooth muscle tissue in the bladder neck to relax the urethra so that urine is discharged smoothly. Therefore, this substance is used for the treatment of diseases in which urine drainage is difficult due to benign prostatic hyperplasia.

Risperidone acts on the central nervous system and is used to treat depression, bipolar disorder, and schizophrenia. Brinzolamide is a carbonic anhydrase inhibitor and is used to treat glaucoma. Clofazimin is used to treat leprosy and tuberculosis, and etoposide is used as an anticancer agent to inhibit the proliferation of tumor cells by binding to DNA and topoisomerase I.

KTKS-palmitate is a substance esterified with palmitic acid to a pentapeptide composed of lysine-threonine-threonine-lysine-serine. This substance is a signaling peptide that promotes the formation of an extracellular matrix in subconfluent fibroblast cell culture. Therefore, foreign substances promote the production of type I and type III collagen and fibronectin and are used as anti-wrinkle and skin aging improving agents. KTKS-palmitate has a molecular weight of 802.05 g/mole and is a kind of small peptide drug with a relatively small molecular size as a peptide substance.

Glutathione is a substance present in all cells and has an antioxidant action that cleans various free radicals that occur in cell metabolism, so it is widely used in food and cosmetics. Sodium risedronate is used as a treatment for osteoporosis by inhibiting the function of osteoclasts. Irinotecan is an anticancer drug that acts antagonistically to topoisomerase I.

Curcumin is a yellow polyphenol component extracted from the root of a plant called Curcuma longa belonging to the ginger family. It has antioxidant, anti-cancer, anti-helicobacterial, anti-inflammatory and immunomodulatory effects, and is widely used in food and cosmetics.

Resveratrol is a polyphenol component extracted from grape seeds and grape skins. It has various actions such as antioxidant, anti-inflammatory, anti-aging, and anti-viral, and is widely used in food and cosmetics.

Quercetin is a polyphenol component contained in onion skins, and it has antioxidant and anti-inflammatory effects like curcumin and resveratrol, and is widely used in food and cosmetics fields.

Lutein is a xanthophyll component found in a lot of spinach, kale, and carrots. Lutein exists in a particularly high concentration in the macula of the retina, and it is known to prevent macular cells from being degenerated by ultraviolet rays, so it is a substance widely used for the prevention and treatment of macular degeneration.

The present invention also provides a composition comprising the lipid nanoparticles into which the physiologically active material is introduced, and a pharmaceutically or food-acceptable carrier. As an example, the composition may be a pharmaceutical composition or a food composition.

In addition, the present invention is the use, prevention, improvement, or therapeutically effective amount of the lipid nanoparticle-bioactive substance complex for the preparation of the pharmaceutical composition or food composition, administering the lipid nanoparticle-bioactive substance complex to the subject. A method of preventing, ameliorating or treating a disease in a subject comprising the subject is provided.

The pharmaceutical composition of the present invention may also include a carrier, diluent, excipient, or a combination of two or more commonly used in biological agents. The pharmaceutically acceptable carrier is not particularly limited as long as it is suitable for delivering the composition in vivo, and for example, saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, or one of these components. It may be a mixture of more than one component. In this case, other conventional additives such as antioxidants, buffers, and bacteriostats may be added as needed. When formulating the composition, it can be prepared using a diluent or excipient such as a filler, extender, binder, wetting agent, disintegrant, surfactant, etc. commonly used. The composition of the present invention may be formulated as an oral preparation or a parenteral preparation. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, troches, and the like, and such solid preparations include at least one excipient in one or more compositions, for example, starch, calcium carbonate, sucrose, It may be prepared by mixing lactose and gelatin. Lubricants such as magnesium stearate and talc may also be added. On the other hand, liquid formulations include suspensions, solutions, emulsions or syrups, and these may include excipients such as wetting agents, sweetening agents, fragrances, and preservatives. Formulations for parenteral administration may include injections such as sterile aqueous solutions, non-aqueous solutions, suspensions, and emulsions. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.

The composition of the present invention may be orally administered or parenterally administered according to a desired method, and parenteral administration is transdermal administration, or intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection. can be selected from

In one embodiment, the composition of the present invention has the characteristics of exhibiting an excellent effect of delivering physiologically active substances through transdermal administration as well as injection administration such as intravenous administration.

The composition according to the present invention is administered in a prophylactically, ameliorated or therapeutically effective amount. This may vary depending on the type of disease, severity, drug activity, sensitivity to drug, administration time, administration route and excretion rate, treatment period, concurrently used drugs, and the like. The composition of the present invention may be administered alone or in combination with other therapeutic agents. When administered in combination, the administration may be sequential or simultaneous. The content of the physiologically active ingredient in the pharmaceutical composition can be appropriately selected by those skilled in the art. The administration may be once a day, or it may be divided into several times.

In another aspect, the present invention provides a method for preparing lipid nanoparticles to which mastic gum or gamma oryzanol is added or a composition comprising the lipid nanoparticles. As a result of seeking a new method beyond the conventional solubilization method, the present inventors introduced a method of melting and mixing mastic gum or gamma oryzanol with heated lipids and oils to produce mastic gum or gamma oryzanol without using harmful organic solvents. A new method that can be miscible with phospholipids and oils was devised.

Specifically, the manufacturing method may include the following steps:

(a) mixing mastic gum or gamma oryzanol with lipid and natural or synthetic oil

(b) as an oil phase, by melting the mixture prepared in step (a) to mix mastic gum or gamma oryzanol in the mixture with phospholipids and natural or synthetic oils and fats, mastic gum or gamma oryzanol, phospholipids, and natural oils or fats or Step of preparing a mixture containing synthetic oil

(c) preparing a lipid nanoparticle composition by cooling a mixture containing mastic gum or gamma oryzanol, lipid, and natural or synthetic oil prepared in step (b)

(d) preparing the award; and

(e) adding and homogenizing the lipid nanoparticle composition prepared in step (c) and the aqueous phase prepared in step (d), or hydrating with the aqueous phase prepared in step (d) and then freeze-drying.

In one aspect, the manufacturing method according to the present invention does not use a harmful organic solvent as an oil phase in step (b), melts the mastic gum or gamma oryzanol, and mixes it with phospholipids and natural or synthetic oils do it with

In one embodiment, the amount of mastic gum or gamma oryzanol added in step (a) is 0.05 wt% to 50 wt%, 0.07 wt% to 47 wt%, 0.09 wt% to 45 wt% or It may be 0.1 wt% to 40 wt%, but is not limited thereto. In addition, the fat may be included in an amount of 1.4 to 2.5 parts by weight, 1.5 parts by weight to 2.0 parts by weight, or 1.6 parts by weight to 1.999 parts by weight, based on 1 part by weight of the phospholipid, but is not limited thereto.

In step (b), the mixture of mastic gum or gamma oryzanol, lipid and natural oil or synthetic oil prepared in step (a) is homogeneously stirred while heating, and then maintained at a predetermined time and a predetermined temperature so that each component is This is the step of mixing well at the molecular level. Although not limited thereto, the heating temperature may be specifically, 90 to 120° C. or 95 to 110° C., but may be appropriately changed and selected. In one embodiment, the mastic or gamma oryzanol is melted and miscible at a molecular level with phospholipids and natural or synthetic oils and fats heated to 110°C. At this time, it can be homogeneously mixed by using a magnetic stirrer or a mechanical stirring device. In addition, the holding time for better mixing may be 5 minutes or more, 10 minutes or more, 5 minutes to 30 minutes, 10 minutes to 30 minutes, or 10 minutes to 25 minutes, but is not limited thereto. The holding temperature may be 100°C to 110°C, 110°C to 120°C, 120°C to 130°C, or 130°C to 140°C, but is not limited thereto.

In addition, after complete mixing through step (b), the mixture prepared as in step (c) is cooled to prepare a lipid nanoparticle composition. At this time, the cooling method is not limited, but may be performed by, for example, air cooling, cold water cooling, or ice cooling. Upon cooling, it is preferably cooled to room temperature to finally prepare a lipid nanoparticle composition. The lipid nanoparticle composition cooled to room temperature may be refrigerated or stored at room temperature as needed.

Also separately, in step (d), an aqueous phase is prepared. As an example of the solvent that can be used as the aqueous phase, water, specifically distilled water, purified water, water for injection, physiological saline, etc. can be used, and in these solvents, a dispersant, an emulsifier, a pH adjuster, an isotonic agent, a preservative, and an antioxidant An aqueous phase may be prepared by adding substances selected from the group.

The step (e) is a step of homogenizing the lipid nanoparticle composition prepared in step (c) and the aqueous phase prepared in step (d), or hydrating the aqueous phase prepared in step (d) and then freeze-drying to be. For the homogenization, a known homomixer, extruder or homogenizer may be used. As an embodiment, the lipid nanoparticle composition in the oil phase container and the aqueous solution in the aqueous phase container are put in a container equipped with a homomixer, extruder or homogenizer, and crushed until the average particle size is, for example, 1 μm or less. Prepare a homogenized lipid nanoparticle dispersion. In this case, an appropriate surfactant may be used to prevent size change after dispersion and ease of dispersion of the lipid nanoparticles. Examples of the surfactant include, but are not limited to, PEG, glycerol, propylene glycol, dipropylene glycol, sorbitan-fatty acid ester, sorbitan-PEG-fatty acid ester, castor oil-PEG derivative, and the like.

In addition, in one aspect, when it is intended to load a fat-soluble substance or a nutrient component as a physiologically active ingredient in the lipid nanoparticles, mixing mastic gum or gamma oryzanol with lipids and fats, which is step (a) of the manufacturing method, It can be loaded by adding the fat-soluble substance or nutrient component to lipids and oils and performing a subsequent process.

In addition, in the case of loading a water-soluble substance or a nutrient component as a physiologically active ingredient in the lipid nanoparticles, when preparing the aqueous phase in step (d) of the manufacturing method, a pH adjuster, an isotonic agent, a preservative, an antioxidant, etc. It can be loaded by adding water-soluble substances or nutrients to the aqueous phase and then proceeding with the process.

In the case of manufacturing lipid nanoparticles or a composition thereof according to the production method of the present invention, there is no toxicity problem due to the use of harmful organic solvents in that lipid nanoparticles can be prepared without using any harmful organic solvents, and There is an advantage in that a separate process is not required to reduce the toxicity problem. Furthermore, in one embodiment according to the present invention, it is confirmed that the dispersion containing the lipid nanoparticles prepared according to the present invention has much better dispersion stability than the dispersion containing the lipid nanoparticles prepared using cholesterol and a harmful organic solvent. did.

In one embodiment, it was confirmed that the lipid nanoparticle dispersion prepared without using any harmful organic solvents according to the present invention showed very good results in the bioavailability test results for transdermal administration and oral administration.

In one embodiment, the dispersion of lipid nanoparticles according to the present invention exhibits a higher zeta potential compared to that without mastic or gamma oryzanol, thereby providing a homogeneous and stable dispersion, thereby improving the dispersion stability of lipid nanoparticles. did.

In one embodiment, it was confirmed that the dispersion of lipid nanoparticles according to the present invention exhibited high encapsulation efficiency (encapsulation rate) of 97% or more for fat-soluble substances and 57.96% for water-soluble substances even when the molecular weight was large.

The manufacturing method according to the present invention is schematically shown in FIG. 1 .

Hereinafter, the present invention will be described in detail by the following examples. However, the following examples are only for illustrating the present invention, and the present invention is not limited thereto. Anything that has substantially the same configuration as the technical idea described in the claims of the present invention and achieves the same operation and effect is included in the technical scope of the present invention.

[Example]

Example 1: Preparation of lipid nanoparticles prepared with mastic gum or gamma oryzanol

Phospholipids (DPPC) and mastic gum or gamma oryzanol are mixed with oil and propylene glycol to form an oil phase. In this case, phospholipids and phospholipids are products extracted from soybeans or egg yolk, and phospholipids having a neutral charge or positive charge may be used. The amount of mastic gum or gamma oryzanol added is between 0.1% and 40% based on the weight of the phospholipid. Glycerin fatty acid ester is preferred for oil and fat, but horse oil, shark oil, light oil (whale oil), krill oil, butter oil or synthetic oil can also be used. Propylene glycol can be used instead of dipropylene glycol or a liquid surfactant. When the temperature reaches 95°C, the phospholipids and mastic gum or gamma oryzanol begin to melt. When mixing is complete, the temperature is maintained at 110° C. for at least 15 minutes so that each component is well mixed at the molecular level. If you want to make a fat-soluble substance or a physiologically active substance into a drug delivery system, you can add the substance at the mixing stage. When the mixing is completed, it is cooled to room temperature to obtain a lipid nanoparticle composition. The lipid nanoparticle composition cooled to room temperature may be refrigerated or stored at room temperature.

Separately from the oil phase, add necessary ingredients such as distilled or purified water, dispersant, emulsifier, isotonic agent, pH adjuster, and preservative to the aqueous phase container and heat to the same temperature as the oil phase container. If you want to make a water-soluble substance into a drug delivery system, you can add the substance to the water phase.

Finally, put the composition in the oil phase container and the solution in the aqueous phase container into a container equipped with a homomixer or homogenizer, crush until the average particle size is 1 micron or less, or hydrate the oil phase with the aqueous phase, then freeze-dried. Prepare lipid nanoparticles.

As an example of lipid nanoparticles prepared using mastic gum or gamma oryzanol, the addition amount of mastic gum or gamma oryzanol was varied from 0.1 wt% to 40 w/w% relative to the weight of phospholipids, and the lipid nanoparticles according to the above method A composition was prepared, and the composition of the lipid nanoparticle composition is shown in Table 1 below.

Composition of Lipid Nanoparticle Composition code DPPC (g) mastic gum or gamma oryzanol (g) glycerine fatty acid ester (g) dipropylene glycol (g) composition 1 5 0.005 99.95 5 composition 2 5 0.05 9.95 5 composition 3 5 One 9.0 5 composition 4 5 2 8.0 5

Example 1-1: Confirmation of zeta potential, particle size and polydispersity index of lipid nanoparticle dispersion prepared with mastic gum or gamma oryzanol

In Example 1, the lipid nanoparticle composition prepared by varying the amount of mastic gum or gamma oryzanol from 0.1 wt% to 40 wt% relative to the phospholipid weight was homogenized in 60 times the amount of distilled water based on the weight to obtain a dispersion, and then zeta The potential, particle size and polydispersity index were measured. The zeta potential was measured 5 times by the electrophoretic light scattering method in an electric field of 25°C and 16 V/cm intensity for Compositions 1 to 4 dispersed in phosphate buffer (pH 6.5), and the average value was obtained. The particle size and polydispersity index were measured 15 times by dynamic light scattering method at a fixed angle of 90 degrees for Composition 1 to Composition 4 dispersed in a phosphate buffer (pH 6.5) prepared at a solid concentration of 0.05% w/w, and the average value was measured 15 times. saved Table 2 shows the zeta potential, particle size, and polydispersity index of Compositions 1 to 4 prepared by varying the amount of mastic gum or gamma oryzanol added from 0.1% to 40% based on the weight of the phospholipid. As shown in Table 2, as the ratio of mastic gum or gamma oryzanol to the weight of phospholipids increased, the zeta potential increased proportionally, and thus the particle size decreased proportionally. It is judged that as the zeta potential increases, a repulsive force is generated between the particles, which does not cause agglomeration and the particle size decreases. The polydispersity index also showed a more uniform distribution as the zeta potential increased and the particle size decreased. In general, it is known that a significant repulsive force is generated between nanoparticles having an absolute value of zeta potential of 25 mV or more to form a stable dispersion without agglomeration. As an example of lipid nanoparticles prepared with mastic gum or gamma oryzanol, the particle size distribution of composition 4 is shown in FIG. 2 .

For reference, Table 3 shows the zeta potential, particle size, and polydispersity index of the lipid nanoparticle dispersion prepared with a composition not using mastic gum in Composition 3 in the manufacturing process of Example 1. As shown in Table 3 , when mastic gum or gamma oryzanol is not added, the zeta potential is -19.5±0.7 mV, the particle size is 585.1±12.7 nm, and the polydispersity index is 0.766±0.050, which is a big difference compared to the case when mastic gum is added. was shown. The polydispersity index is an index that can predict whether a homogeneous and stable dispersion is a homogeneous and stable dispersion when it is less than 0.3.

The lipid nanoparticle dispersion prepared without mastic gum or gamma oryzanol had a polydispersity index of 0.766, but when mastic gum or gamma oryzanol is used, it becomes smaller depending on the amount. It was determined that gamma oryzanol could improve the dispersion stability of lipid nanoparticles.

Zeta potential, particle size and polydispersity index of lipid nanoparticle dispersion prepared without mastic gum or gamma oryzanol zeta potential (mV) particle size (nm) Polydispersity index -19.5±0.7 585.1±12.7 0.766±0.050

Examples 1-2: Electron transmission microscope confirmation of lipid nanoparticle dispersion prepared with mastic gum

As an example of lipid nanoparticles prepared using mastic gum , composition 4 was homogenized in 60 times the amount of distilled water based on weight to make a dispersion, and then photographed with an electron transmission microscope (JEM-2100F) to determine the structure of the lipid nanoparticles. I looked. The experimental method is as follows. 100 mg of Composition 4 was added to 10 ml of distilled water and then homogenized at 24,000 rpm for 2 minutes (IKA homogenizer) to prepare a lipid nanoparticle dispersion. A drop of this dispersion was placed on a carbon-coated copper grid, and distilled water used as a dispersion medium was allowed to dry naturally, and the remaining water that did not dry even after 5 minutes was sucked out with a filter paper. A drop of Phosphotungstic acid 1% solution was dropped on the dried sample, and when it was completely dried, it was mounted on the stage of the microscope and images were taken at 100 kV. As a result, as shown in FIG. 3 , it was found that most of the nanoparticles were spherical, and the size was consistent with the results of Table 2 . Although some particles were agglomerated and lumps were observed, most of the particles were not agglomerated and existed individually.

Example 1-3: Dispersion stability of lipid nanoparticles prepared with mastic gum or gamma oryzanol

In Example 1, the lipid nanoparticle composition prepared by changing the amount of mastic gum or gamma oryzanol from 0.1% to 40% by weight relative to the weight of the phospholipid was homogenized in 60 times the amount of distilled water based on the weight to obtain a dispersion, and then dispersed Stability was measured and shown in FIG. 4 . Dispersion stability was evaluated by measuring the particle size and zeta potential for 4 weeks while storing the dispersion at 25±2° C.

As shown in FIG. 4 , the average particle size of the lipid nanoparticles of Composition-1 prepared using only 0.1% of mastic gum compared to phospholipids was 522.1±10.9 nm at first, but 759.2±36.7 nm at the 1st week, and 1067.4± at the 4th week. It increased significantly to 120.4 nm. On the other hand, the average particle size of the lipid nanoparticles of Composition-3 prepared using mastic gum in 20% by weight relative to the phospholipids was 303.4±15.6 nm at the beginning, and there was little change to 325.6±50.4 nm even at the 4th week. In particular, in the case of Composition-4 using 40% mastic gum compared to phospholipids, the average particle size was 300 nm or less even after 4 weeks of storage, indicating very stable dispersibility. Even when gamma oryzanol was used, it showed the same tendency as when mastic gum was used. The reason why the dispersibility of the lipid nanoparticles is stabilized is because the absolute value of the zeta potential is increased and maintained constant due to the use of mastic gum or gamma oryzanol of the nanoparticles.

Example 1-4: Confirmation of properties of receptor compartment liquid after transdermal permeation test of lipid nanoparticle dispersion prepared with mastic gum or gamma oryzanol

Compositions 1 and 4 , prepared using mastic gum or gamma oryzanol in Example 1, were homogenized with 60 times the weight of distilled water and applied to the donor compartment of the Franz diffusion cell, and then 24 hours later, the receptor compartment solution It is shown in Figure 5 by taking a picture of the properties of. The effective diffusion area of the Franz diffusion cell was 2.087 cm ² , and the skin used for the test was purchased from pig skin within 24 hours of slaughter. Pig skin was placed in a thickness of about 1 mm by removing subcutaneous fat so that the stratum corneum layer was directed toward the donor compartment and the dermis layer was directed toward the receptor compartment.

As a result, the receptor compartment properties of the composition 1 dispersion were only slightly turbid compared to the time at which the test was started, but in the case of the composition 4 dispersions, the receptor compartment properties showed a very cloudy color unlike the composition 1 dispersion. This indicates that lipid nanoparticles in the dispersion in the donor compartment penetrated the skin and migrated to the receptor compartment. This result is evidence that the lipid nanoparticles in the dispersion of Composition 4 penetrated the skin, which is because the average particle size of the lipid nanoparticles in the dispersion is very small, about 250 nm, and the polydispersity index is less than 0.3 because it is a very homogeneous dispersion. to be.

Example 2 : Comparison with lipid nanoparticles prepared using cholesterol

The inventors have found that lipid nanoparticles prepared by replacing cholesterol with mastic gum or gamma oryzanol form a stable dispersion. In this experiment, the sample using mastic gum was used as composition 3 , and the sample using cholesterol was prepared using the same weight of cholesterol and other raw materials in composition 3 . For the preparation of the cholesterol-using sample, it was separately put into 10 ml of ethanol to dissolve cholesterol, completely dissolved by heating to 70° C., and then mixed with other raw materials, and then the ethanol was evaporated with a dark evaporator. After the experimental procedure, the zeta potential, particle size, and polydispersity index of the lipid nanoparticle dispersion were measured according to the method of Example 1-1 . The results are shown in Table 4 .

As shown in Table 4 , the sample prepared using mastic gum or gamma oryzanol was superior to the sample prepared using cholesterol in all areas such as zeta potential, particle size, and polydispersity index. In particular, when cholesterol was replaced with mastic gum or gamma oryzanol, the absolute value of the zeta potential almost doubled. Considering that the zeta potential is an important variable that can predict the dispersion stability of nanoparticles, the excellent stability of the composition 3 dispersion prepared by replacing cholesterol with mastic gum or gamma oryzanol is a result of the existing lipid nanoparticles using cholesterol. It can be seen that this is because the absolute value of the zeta potential is significantly increased compared to that ( see composition 2, composition 3, and composition 4 in FIG . 4 ).

Comparison with lipid nanoparticles prepared using cholesterol code zeta potential (mV) particle size (nm) polydispersity index Sample using mastic gum (composition 3) -56.1±0.8 303.4±15.6 0.257±0.002 Sample using gamma oryzanol (Composition 3) -45.6±0.3 262.7±24.3 0.206±0.001 Cholesterol Use Sample -29.58±0.9 591.76±27.3 0.606±0.050

Example 3: Preparation of lipid nanoparticles encapsulated in fat-soluble substances

An experiment was performed to encapsulate a fat-soluble material as a model material in lipid nanoparticles prepared using mastic gum or gamma oryzanol. Since the fat-soluble substance is hardly soluble in water but soluble in oil, it was put in an oil phase with phospholipids, mastic gum or gamma oryzanol, fats and oils, etc., mixed and mixed, and encapsulated in lipid nanoparticles. The manufacturing process is the same as in Example 1 except that the fat-soluble substance is added to the oil phase in the mixing step with other raw materials. Table 5 shows the composition and amount of raw materials used to prepare lipid nanoparticles encapsulated with fat-soluble substances.

Composition of lipid nanoparticle composition encapsulated in fat-soluble substances code DPPC (g) mastic gum　 or gamma oryzanol (g) glycerine fatty acid ester (g) dipropylene glycol (g) Fat-soluble substances (g) composition 5 5 0.005 9.495 5 0.5 composition 6 5 0.05 9.45 5 0.5 composition 7 5 One 8.5 5 0.5 composition 8 5 2 7.5 5 0.5 Fat-soluble substances: dutasteride, finasteride, ketoprofen, lidocaine, minoxidil, docetaxel, tamsulosine, KTTKS-palmitate, curcumin, resveratrol, quercetin, lutein, risperidone, brinzolamide, clofazimin, eto forsyth

Example 3-1: Transdermal permeation test of lipid nanoparticle dispersions encapsulated with fat-soluble substances

Various lipid-soluble substances with the compositions shown in Table 5 of Example 3 were used as model substances, mounted on lipid nanoparticles, and the skin permeation phenomenon of the substances was measured.

The transdermal permeation test was measured using a Franz diffusion cell with an effective diffusion area of 2.087 cm ² . The skin used for the test was used after purchasing pig skin less than 24 hours after slaughter, removing the subcutaneous fat and making it about 1 mm thick. The stratum corneum layer of the skin was oriented towards the donor compartment and the dermis layer was oriented towards the receptor compartment.

In the donor compartment, 100 mg of the lipid nanoparticle composition of Composition 5 to Composition 8 loaded with the material prepared in Example 3 was placed in 6 ml of distilled water (60-fold dilution) and homogenized at 24,000 rpm for 2 minutes (IKA homogenizer), followed by 0.5 dispersion. ml was taken and added. To prevent the dispersion in the donor compartment from evaporating, the upper opening was covered with parafilm. The receptor compartment was filled with 11 ml of distilled water, and a donor blood was installed between the donor compartment and the receptor compartment. In the Franz diffusion cell test apparatus, the temperature was kept constant while circulating water at 37°C using a water bath.

Samples in the receptor compartment were collected at set time intervals (1, 2, 3, 6, 12, 24 hours) at 500 μl and stored in a deep freezer (-80°C). The volume reduced due to sampling was maintained by injecting the same volume of distilled water each time to keep the volume of the receptor compartment constant. When the last sample was collected, the concentration of the substance was quantitatively analyzed by HPLC without delay.

The HPLC analysis conditions of each material are shown in Table 6 below.

The results of the transdermal permeation test using pig skin of lipid nanoparticles encapsulated with dutasteride are shown in FIG. 6 . As shown in the figure, in the case of composition-5 (mastic gum) containing dutasteride, the transdermal permeability was only 12.54±6.24% of the amount of dutasteride applied to the donor compartment even after 24 hours, but composition-8 containing dutasteride (each stick gum) , it is 76.59±6.26%, indicating that more than three quarters of the applied amount has been transmitted. Also, the cumulative transdermal permeability increased proportionally with the content of mastic gum or gamma oryzanol.

The results of the transdermal permeation test using pig skin of lipid nanoparticles encapsulated with finasteride are shown in FIG. 7 . As shown in the figure, in the case of finasteride-containing composition-5 (gamma oryzanol) , the transdermal permeability was only 16.87±6.01% for 24 hours, but in the case of finasteride-containing composition-8 (gamma oryzanol) , it was 56.87±1.36%, which increased about three times. Also, the cumulative transdermal permeability increased proportionally with the content of mastic gum or gamma oryzanol.

The results of the transdermal permeation test using pig skin of lipid nanoparticles encapsulated with ketoprofen are shown in FIG. 7 . As shown in FIG. 7, in the case of ketoprofen-containing composition-5 (gamma oryzanol) , the transdermal permeability was only 53.65±2.68% for 24 hours, but in the case of ketoprofen-containing composition-7 8 (gamma oryzanol) , it was 86.97±8.29%. increased by 1.6 times. In addition, the cumulative transdermal permeability showed a tendency to increase proportionally with the content of mastic gum or gamma oryzanol.

The results of the transdermal permeation test using pig skin of lipid nanoparticles encapsulated with lidocaine are shown in FIG. 9 . As shown in FIG. 9 , in the case of lidocaine-containing composition-5 (gamma oryzanol) , the transdermal permeability was 50.27±5.26% for 24 hours, but in the case of lidocaine-containing composition-8 (gamma oryzanol) , almost 100% was transmitted as 98.57±5.27%. indicated. In addition, the cumulative transdermal permeability showed a tendency to increase proportionally with the content of mastic gum or gamma oryzanol.

The results of the transdermal permeation test using pig skin of lipid nanoparticles encapsulated with minoxidil are shown in FIG. 10 . As shown in FIG. 10 , the transdermal permeability for 24 hours was 25.81±4.59% for the minoxidil- containing composition-5 (mastic gum) , but 70.84±1.99% for the minoxidil-containing composition-8 (mastic gum) . In addition, the cumulative transdermal permeability showed a tendency to increase proportionally with the content of mastic gum or gamma oryzanol.

The results of the transdermal permeation test using pig skin of lipid nanoparticles encapsulated with docetaxel are shown in FIG. 11 . As shown in FIG. 11 , in the case of docetaxel-containing composition-5 (gamma oryzanol) , the transdermal permeability was 13.05±5.37% for 24 hours, but in the case of docetaxel-containing composition-8 (gamma oryzanol) , it was 53.54±5.63%. In addition, the cumulative transdermal permeability showed a tendency to increase proportionally with the content of mastic gum or gamma oryzanol.

The results of the transdermal permeation test using pig skin of lipid nanoparticles encapsulated with tamsulosin are shown in FIG. 12 . As shown in FIG. 12 , the transdermal permeability for 24 hours was 11.70±6.25% in the case of tamsulosine -containing composition-5 (mastic gum) , but 47.65±5.67% in the case of tamsulosine -containing composition-8 (mastic gum) . It was. In addition, the cumulative transdermal permeability showed a tendency to increase proportionally with the content of mastic gum or gamma oryzanol.

The results of the transdermal permeation test using pig skin of lipid nanoparticles encapsulated with KTKS-palmitate are shown in FIG. 13 . As shown in FIG. 13 , in the case of KTTKS-palmitate-containing composition-5 (mastic gum) , the transdermal permeability was 14.30±2.92% for 24 hours, but in the case of KTKS-palmitate-containing composition-8 (mastic gum) , it was 3.4 to 48.65±5.64%. appeared to be improved by a factor of two. In addition, the cumulative transdermal permeability showed a tendency to increase proportionally with the content of mastic gum or gamma oryzanol.

The results of the transdermal permeation test using pig skin of lipid nanoparticles encapsulated with curcumin are shown in FIG. 14 . As shown in FIG. 14 , in the case of curcumin-containing composition-5 (gamma oryzanol) , the transdermal permeability was 20.23±4.19% for 24 hours, but in the case of curcumin-containing composition-8 (gamma oryzanol) , it was 65.87±8.06%. In addition, the cumulative transdermal permeability showed a tendency to increase proportionally with the content of mastic gum or gamma oryzanol.

The results of the transdermal permeation test using pig skin of lipid nanoparticles encapsulated with resveratrol are shown in FIG. 15 . As shown in FIG. 15 , the transdermal permeation amount for 24 hours was 18.24±5.97% for the resveratrol-containing composition -5 (gamma orizanol), but 79.67±5.79% for the resveratrol-containing composition-8 (gamma oryzanol) . In addition, the cumulative transdermal permeability showed a tendency to increase proportionally with the content of mastic gum or gamma oryzanol.

The results of the transdermal permeation test using pig skin of lipid nanoparticles encapsulated with quercetin are shown in FIG. 16 . As shown in FIG. 16 , the transdermal permeability of quercetin-containing composition-5 (mastic gum) for 24 hours was 25.12±6.68%, but the quercetin-containing composition-8 (mastic gum) showed 85.97±7.24%. In addition, the cumulative transdermal permeability showed a tendency to increase proportionally with the content of mastic gum or gamma oryzanol.

The results of the transdermal permeation test using pig skin of lipid nanoparticles encapsulated with lutein are shown in FIG. 17 . As shown in FIG. 17 , the transdermal permeability of Lutein-containing composition-5 (gamma oryzanol) was 8.67±2.19% for 24 hours, but 65.87±5.36% of the lutein-containing composition-8 (gamma oryzanol) . In addition, the cumulative transdermal permeability showed a tendency to increase proportionally with the content of mastic gum or gamma oryzanol.

The results of the transdermal permeation test using pig skin of lipid nanoparticles encapsulated with risperidone are shown in FIG. 18 . As shown in FIG. 18 , in the case of risperidone-containing composition-5 (gamma oryzanol) , the transdermal permeability was 7.27±2.10% for 24 hours, but in the case of risperidone-containing composition-8 (gamma orizanol) , it was 35.28±7.46%. In addition, the cumulative transdermal permeability showed a tendency to increase proportionally with the content of mastic gum or gamma oryzanol.

The results of the transdermal permeation test using pig skin of lipid nanoparticles encapsulated with brinzolamide are shown in FIG. 19 . As shown in FIG. 19 , the transdermal permeation rate for 24 hours was 14.20±3.65% in the case of brinzolamide -containing composition-5 (gamma oryzanol) , but 52.64±4.20% in the case of brinzolamide -containing composition-8 (gamma oryzanol) . In addition, the cumulative transdermal permeability showed a tendency to increase proportionally with the content of mastic gum or gamma oryzanol.

The results of the transdermal permeation test using pig skin of lipid nanoparticles encapsulated with clofazimin are shown in FIG. 20 . As shown in FIG. 20 , in the case of clofazimin -containing composition-5 (gamma oryzanol) , the transdermal permeability was only 6.86±2.99% for 24 hours, but in the case of clofazimin -containing composition-8 (gamma orizanol) , it was 31.24±6.69%. It was. In addition, the cumulative transdermal permeability showed a tendency to increase proportionally with the content of mastic gum or gamma oryzanol.

The results of the transdermal permeation test using pig skin of lipid nanoparticles encapsulated with etoposide are shown in FIG. 21 . As shown in FIG. 21 , the transdermal permeability for 24 hours was only 20.68±3.07% in the case of etoposide-containing composition-5 (gamma oryzanol) , but 65.00±8.57% in the case of etoposide-containing composition-8 (gamma oryzanol) . In addition, the cumulative transdermal permeability showed a tendency to increase proportionally with the content of mastic gum or gamma oryzanol.

HPLC analysis conditions for fat-soluble substances substance name column Column temperature (℃) mobile phase Flow rate (ml/min) Injection volume (μl) UV detector wavelength (nm) holding time (min) dutasteride Inertsil ODS-3 (4.6×150 mm, 5㎛) 30 Acetonitrile: 1% distilled water containing ortho-phosphoric acid (70:30 v/v) 0.8 25 210 7.9 finasteride Inertsil ODS-3 (4.6×150 mm, 5㎛) 25 Acetonitrile: 1% distilled water containing ortho-phosphoric acid (70:30 v/v) 0.4 25 210 5.9 ketoprofen Inertsil ODS-3 (4.6×150 mm, 5㎛) 25 Acetonitrile: 1% distilled water containing ortho-phosphoric acid (65:35 v/v) 0.4 25 260 7.9 lidocaine Inertsil ODS-3 (4.6×150 mm, 5㎛) 25 Acetonitrile: 1% distilled water containing ortho-phosphoric acid (15:85 v/v) 0.5 25 235 7.1 minoxidil Inertsil ODS-3 (4.6×150 mm, 5㎛) 25 Acetonitrile: 0.1% triethanolamine-containing distilled water mixture (60:40 v/v) 0.6 25 285 6.2 docetaxel Inertsil ODS-3 (4.6×150 mm, 5㎛) 25 Acetonitrile: Distilled water mixture containing 0.5% ortho-phosphoric acid (25:75 v/v) 0.5 25 210 6.2 tamsulosin Inertsil ODS-3 (4.6×150 mm, 5㎛) 25 Acetonitrile: 1% distilled water containing ortho-phosphoric acid (68:32 v/v) 0.4 25 280 7.6 KTKS-palmitate Inertsil ODS-3 (4.6×150 mm, 5㎛) 25 Acetonitrile containing 0.1% trifluoroacetic acid: A mixture of distilled water containing 0.1% trifluoroacetic acid (50:50 v/v) 0.4 25 214 6.1 curcumin Prodigy ODS-3 (4.6×150 mm, 5㎛) 25 Acetonitrile: 2% acetic acid-containing distilled water mixture (70:30 v/v) 0.4 25 420 7.1 resveratrol Inertsil ODS-3 (4.6×150 mm, 5㎛) 20 Acetonitrile: 2% acetic acid-containing distilled water mixture (50:50 v/v) 0.45 25 346 7.0 quercetin Inertsil ODS-3 (4.6×150 mm, 5㎛) 20 Acetonitrile: 2% acetic acid-containing distilled water mixture (50:50 v/v) 0.4 25 346 8.2 lutein Inertsil ODS-3 (4.6×150 mm, 5㎛) 25 Acetonitrile:methanol:methylene chloride mixture (40:40:20 v/v) 1.0 25 445 6.9 risperidone Inertsil CN-3 (4.6×250 mm, 5㎛) 25 Acetonitrile: 0.2% triethanolamine-containing distilled water mixture (45:55 v/v) 0.5 30 280 8.5 brinzolamide Prodigy ODS-3 (4.6×250 mm, 5㎛) 25 Acetonitrile:methanol:phosphate buffer (pH 6.0) (15:40:45 v/v) 1.0 25 254 3.9 clofazimine Inertsil ODS-3 (4.6×250 mm, 5㎛) 30 Methanol:acetate buffer (pH3.0) (75:25 v/v) 1.0 25 286 10.2 etoposide Inertsil ODS-3 (4.6×150 mm, 5㎛) 25 Acetonitrile:acetate buffer (pH 4.6) (40:60 v/v) 2.0 25 285 7.8

Example 4 : Preparation of lipid nanoparticles encapsulated with water-soluble substances

An experiment was performed to prepare a dispersion by encapsulating a water-soluble material as a model material in lipid nanoparticles prepared using mastic gum or gamma oryzanol. Since the water-soluble substance is hardly soluble in oil and is well soluble in water, it was placed in an aqueous phase and encapsulated in lipid nanoparticles. The manufacturing process is the same as in Example 1 except that a water-soluble substance is put in an aqueous phase, hydrated, and then freeze-dried. The composition and amount of the raw material used for the preparation of the lipid nanoparticles encapsulated in the water-soluble substance are shown in Table 7.

Composition of lipid nanoparticles encapsulated with water-soluble substances code DPPC (g) mastic gum or gamma oryzanol (g) glycerine fatty acid ester　 (g) dipropylene glycol (g) Water-soluble substances (g) Purified water
(g) composition 9 5 0.005 9.495 5 see footnote 20 composition 10 5 0.05 9.45 5 see footnote 20 composition 11 5 One 8.5 5 see footnote 20 composition 12 5 2 7.5 5 see footnote 20 Water-soluble substances: ascorbic acid 0.5 g, epidermal growth factor 0.01 g, glutathione 8 g, risedronate sodium 4.5 g, irinotecan 4 g

Example 4-1 : Percutaneous penetration test of lipid nanoparticles encapsulated with water-soluble substances

In Table 7 of Example 4 , water-soluble substances such as ascorbic acid, epidermal growth factor, glutathione, risedronate sodium, and irinotecan were used as model substances and encapsulated in lipid nanoparticles, and the skin permeation phenomenon of the substances was measured.

Ascorbic acid is a powerful antioxidant and vitamin C required for the activation of various enzymes, including enzymes involved in collagen synthesis.

Epidermal growth factor consists of 53 amino acids and is a type of peptide drug with a molecular weight of 6400 g/mole. It is a signaling peptide that binds to the epidermal growth factor receptor (EGFR) of the cell membrane and induces the growth and differentiation of epidermal cells. Therefore, this substance is used for the purpose of improving skin elasticity and helping to improve wrinkles by promoting the generation of epidermal cells.

Glutathione is a peptide compound composed of three amino acids: glutamic acid, cysteine, and glycine. It is an antioxidant that converts cellular damage-causing substances such as free radicals, peroxides, and lipid peroxides into harmless substances. Glutathione is a useful substance with antioxidant and anti-aging effects, but has a very low absorption rate through the digestive tract and skin.

Sodium risedronate is a bisphosphonate compound and has a high affinity with hydroxyapatite, which is a component of bone, and is dormant in the bone. However, this drug has a problem of very low bioavailability because the absorption rate is only less than 1% when administered orally.

Irinotecan is a compound belonging to a plant alkaloid, which inhibits topoisomerase I and causes damage to RNA and DNA, so it is a drug used as an anticancer agent for various types of cancer such as rectal cancer, colon cancer, stomach cancer, small cell lung cancer, and non-small cell lung cancer. However, since this drug has no specificity for cancer cells, there is a problem in that it is cytotoxic to normal cells.

The transdermal permeation test was carried out in the same manner as in Example 3-1 by dispersing the lipid nanoparticles prepared with the composition shown in Table 7 in an appropriate amount of distilled water, and then using a Franz diffusion cell having an effective diffusion area of 2.087 cm ² . When the last sample was collected in the transdermal permeation test, the concentration of the substance was quantitatively analyzed under the HPLC conditions shown in Table 8 without delay.

The results of the transdermal permeation test using pig skin of lipid nanoparticles encapsulated with ascorbic acid are shown in FIG. 22 . 22 , in the case of ascorbic acid-containing composition-9 (mastic gum) prepared using mastic gum, the transdermal permeability was 13.25±9.26% for 24 hours, but ascorbic acid-containing composition-12 prepared using mastic gum In the case of (mastic gum) , it was found to be 51.26±5.65%, which was improved by 3.9 times. In addition, the cumulative transdermal permeability showed a tendency to increase proportionally with the content of mastic gum or gamma oryzanol.

The results of the transdermal permeation test using pig skin of lipid nanoparticles encapsulated with epidermal growth factor are shown in FIG. 23 . As shown in FIG. 23 , in the case of composition-9 (mastic gum) containing epidermal growth factor prepared using mastic gum, the transdermal permeability was 5.98±2.36% for 24 hours, but composition-12 containing epidermal growth factor prepared using mastic gum In the case of (mastic gum) , it was 34.26±2.67%, which was improved by 5.7 times. In addition, the cumulative transdermal permeability showed a tendency to increase proportionally with the content of mastic gum or gamma oryzanol.

The results of the transdermal permeation test using pig skin of lipid nanoparticles encapsulated with glutathione are shown in FIG. 24 . As shown in FIG. 24 , in the case of glutathione-containing composition-9 (mastic gum) prepared using mastic gum, the transdermal permeability was 2.16±1.20% for 24 hours, but glutathione-containing composition-12 (mastic gum) prepared using mastic gum ) was 23.27±5.64%, which was an improvement of 10.8 times. On the other hand, in the case of glutathione-containing composition-9 (gamma-orizanol) prepared using gamma-orizanol, the transdermal permeability was 2.34±3.67% for 24 hours, but in the case of glutathione-containing composition-12 (gamma-orizanol) prepared using gamma-orizanol, 48.97± It was found to be improved by 20.9 times to 7.26%. In addition, the cumulative transdermal permeability showed a tendency to increase proportionally with the content of mastic gum or gamma oryzanol.

The results of the transdermal permeation test using the pig skin of lipid nanoparticles encapsulated with risedronate are shown in FIG. 25 . As shown in FIG. 25 , in the case of risedronate-containing composition-9 (mastic gum) prepared using mastic gum, the transdermal permeability was 2.65±1.30% for 24 hours, but risedronate-containing composition-12 prepared using mastic gum In the case of (mastic gum), 10.26±2.97% was found to be improved by 3.9 times. On the other hand, in the case of risedronate-containing composition-9 (gamma-orizanol) prepared using gamma-oryzanol, the transdermal permeability was 5.28±3.67% for 24 hours, but the risedronate-containing composition-12 (gamma-orizanol) prepared using gamma-orizanol was case, it was 37.87±7.26%, an improvement of 7.2 times. In addition, the cumulative transdermal permeability showed a tendency to increase proportionally with the content of mastic gum or gamma oryzanol.

The results of the transdermal permeation test using pig skin of lipid nanoparticles encapsulated with irinotecan are shown in FIG. 26 . As shown in FIG. 26 , in the case of irinotecan-containing composition-9 (mastic gum) prepared using mastic gum, the transdermal permeability was 3.78±1.26% for 24 hours, but irinotecan-containing composition-12 (mastic gum) prepared using mastic gum ), 20.64±4.67%, which was about 5.5 times improved. On the other hand, in the case of irinotecan-containing composition-9 (gamma-orizanol) prepared using gamma-orizanol, the transdermal permeability was 12.87±5.24% for 24 hours, but in the case of irinotecan-containing composition-12 (gamma-orizanol) prepared using gamma-orizanol, 44.68± It was shown that it was improved by 3.5 times to 4.24%. In addition, the cumulative transdermal permeability showed a tendency to increase proportionally with the content of mastic gum or gamma oryzanol.

HPLC analysis conditions for water-soluble substances substance name column Column temperature (℃) mobile phase Flow rate (ml/min) Injection volume (μl) UV detector wavelength (nm) holding time (min) ascorbic acid Inertsil ODS-3 (4.6×150 mm, 5㎛) 25 Acetonitrile: 0.1% acetic acid-containing distilled water mixture (45:55 v/v) 0.45 25 250 5.9 epidermal growth factor Inertsil ODS-3 (4.6×150 mm, 5㎛) 20 Acetonitrile: 1% distilled water containing ortho-phosphoric acid (26:74 v/v) 0.4 25 214 5.6 glutathione Inertsil CN-3 (4.6×250 mm, 5㎛) 35 Distilled water containing 0.1% ortho-phosphoric acid 0.3 35 192 10.5 risedronate Inertsil CN-3 (4.6×150 mm, 5㎛) 35 Distilled water containing 0.1% triethanolamine 0.5 35 260 2.2 irinotecan Synergi Polar-RP 80A (4.6×250 mm, 4㎛) 25 Acetonitrile:0.1% trifluoroacetic acid (gradient from 30:70 to 70:30 over 13 min) 0.1 25 372 7.5

Example 4-2 : Absorption test for oral administration of lipid nanoparticle composition encapsulated with water-soluble substances

The absorption test after oral administration of the lipid nanoparticle composition encapsulated with glutathione, a water-soluble substance, was conducted using 8-week-old male Sprague-Dawley rats. Glutathione-containing composition-12 (gamma oryzanol) was homogeneously dispersed in physiological saline and then orally administered as glutathione at a dose of 50 mg/Kg, and then the concentration of glutathione absorbed into the blood was measured (n=5). To the control group, a sample prepared by dissolving pure glutathione powder in physiological saline to have the same concentration as the sample administered to the test drug group was administered. The concentration of glutathione in the blood sample was quantitatively analyzed under the HPLC conditions shown in Table-8 . The concentration of glutathione absorbed into the blood was defined as the increased concentration compared to the glutathione concentration (baseline concentration) measured before administration of the test drug or control drug. As shown in FIG. 27 , the solution obtained by dissolving glutathione powder in physiological saline had an increased blood glutathione concentration of 6.27±5.64 μg/ml, which was hardly absorbed considering the standard deviation. This result confirms the previously known fact that glutathione is hardly absorbed into the blood through oral administration. On the other hand, in the case of glutathione-containing composition-12 (gamma oryzanol) disclosed in the present invention, the increase in blood glutathione concentration was 29.63±6.67 μg/ml, indicating that it was clearly absorbed into the blood. This result shows that the glutathione molecules encapsulated in Composition-12 (gamma oryzanol) exist as lipid nanoparticles, unlike glutathione molecules in a general solution state, and due to this, that is, due to the properties of lipid nanoparticles, they cannot pass through enterocytes. means there is

Example 5 : Confirmation of encapsulation efficiency of lipid nanoparticle dispersion prepared with mastic gum or gamma oryzanol

The encapsulation efficiency of the material encapsulated in the lipid nanoparticles in the lipid nanoparticle dispersion prepared in Example 3 was measured by the following method.

In the case of a fat-soluble substance, 100 mg of the lipid nanoparticle composition loaded with the substance according to the amount shown in Composition 8 of Table 5 was placed in 6 ml of distilled water (60-fold dilution), homogenized at 24,000 rpm for 2 minutes, and then 1 ml of the dispersion was added to Amikon Ultra- Centrifugation was performed at 4,000x g for 30 minutes at 4°C using a 4 filter (MWCO 10,000 g/mol). In this process, the unencapsulated material passes through the Amikon Ultra-4 Filter filter. 50 μl of a sample that passed through the Amikon Ultra-4 Filter filter was taken and quantitatively analyzed under HPLC conditions used for quantitative analysis of each substance mentioned in Example 3 . The column and mobile phase were performed under the same conditions used for quantitative analysis of each material.

In the case of water-soluble substances, the lipid nanoparticles prepared according to the amount shown in Composition 12 of Table 7 were again dispersed in 20 g of purified water, and then 100 mg was added to 6 ml of aqueous phase (60-fold dilution), followed by the same method as in the case of fat-soluble substances. Since the unencapsulated material passes through the Amikon Ultra-4 Filter filter, the sample passing the filter was taken, and then the encapsulation efficiency was measured under HPLC conditions used for quantitative analysis of each material mentioned in Example 4 .

The encapsulation efficiency was calculated by Equation 1 below.

[Equation 1]

Encapsulation efficiency (%) = 100×(D _total D _unentrapped )/D _total

D _total : Weight of substance in lipid nanoparticle dispersion (mcg)

D _unentrapped : Weight (mcg) of material passed through Amikon Ultra-4 Filter (MWCO 10,000 g/mol)

As shown in Table 9 , the encapsulation efficiency of the lipid nanoparticle dispersion prepared with mastic gum or gamma oryzanol was 97% or more for the fat-soluble material. In the case of water-soluble substances, epidermal growth factor, a substance with a large molecular weight, showed a relatively high encapsulation efficiency of 57% or more, and ascorbic acid with a low molecular weight showed a very high encapsulation efficiency of 74.37%. In the case of glutathione, risedronate sodium, and irinotecan, more than 95% and close to 100% were encapsulated.

Claims (13)

Lipid nanoparticles containing mastic gum or gamma oryzanol in lipids. According to claim 1, wherein the lipid is dilauroyl phosphatidylethanolamine (dilauroyl phosphatidylethanolamine), dimyristoyl phosphatidylethanolamine (dimyristoyl phosphatidylethanolamine), dipalmitoyl phosphatidylethanolamine (dipalmitoyl phosphatidylethanolamine), distearoyl ethanol Amine (distearoyl phosphatidylethanolamine), dioleoyl phosphatidylethanolamine (dioleoyl phosphatidylethanolamine), dilinoleoyl phosphatidylethanolamine (dilinoleoyl phosphatidylethanolamine), 1-palmitoyl-2-oleoyl phosphatidylethanolamine (1-palmitoyl-2-oleoylethanolamine) ), 1,2-diphytanoyl-3-sn-phosphatidylethanolamine (1,2-diphytanoyl-3-sn-phosphatidylethanolamine), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine, DOPE), dilauroyl phosphatidylcholine, dimyristoyl phosphatidylcholine, dipalmitoyl phosphatidylcholine, dipalmitoyl phosphatidylcholine Il phosphatidylcholine (distearoyl phosphatidylcholine), dioleoyl phosphatidylcholine (dioleoyl phosphatidylcholine), dilinoleoyl phosphatidylcholine (dilinoleoyl phosphatidylcholine), 1-palmitoyl-2-oleoyl phosphatidylcholine (1-palmitoyl-2-oleoyl-2-oleoyl) -Dipalmitoyl-sn-glycero-3-phosphatidylcholine (1,2-dipalmitoyl-sn-glycero-3-phosphochol) ine, DPPC) 1,2-diphytanoyl-3-sn-phosphatidylcholine (1,2-diphytanoyl-3-sn-phosphatidylcholine), 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine (1, 2-dioleoyl-sn-glycero-3-phosphatidylcholine (DOPC), 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (1,2-dimyristoyl-sn-glycero-3-phosphocholine, DMPC), 1,2-distearoyl-sn-glycero-3-phosphatidylcholine (1,2-distearoyl-sn-glycero-3-phosphocholine, DSPC), dipalmitoyl-phosphatidylglycerol (DPPG) dilau dilauroyl phosphatidic acid, dimyristoyl phosphatidic acid, dipalmitoyl phosphatidic acid, distearoyl phosphatidic acid, diol dioleoyl phosphatidic acid, dilinoleoyl phosphatidic acid, 1-palmitoyl-2-oleoyl phosphatidic acid, and 1, 2-diphytanoyl-3-sn-phosphatidic acid (1,2-diphytanoyl-3-sn-phosphatidic acid) comprising at least one selected from the group consisting of, lipid nanoparticles. The lipid nanoparticles according to claim 1, wherein the mastic gum or gamma oryzanol is included in an amount of 0.05 to 50% by weight based on the weight of the lipid. According to claim 1, which further comprises fats and oils, lipid nanoparticles. The lipid nanoparticles according to claim 4, wherein the oil is vegetable oil or animal oil. The lipid nanoparticles of claim 4, wherein the oil is included in an amount of 1.4 to 2.5 parts by weight based on 1 part by weight of the lipid. The method of claim 1, further comprising at least one surfactant selected from the group consisting of PEG, glycerol, propylene glycol, dipropylene glycol, sorbitan-fatty acid ester, sorbitan-PEG-fatty acid ester, and castor oil-PEG derivative. That is, lipid nanoparticles. According to claim 1, having a particle diameter of 10 to 500nm, lipid nanoparticles. The lipid nanoparticles according to claim 1 or a physiologically active material introduced into a composition comprising the lipid nanoparticles,
In this case, the physiologically active material is a composition for delivery of a physiologically active material that is introduced into the composition comprising the lipid nanoparticles or lipid nanoparticles. The composition of claim 9, wherein the physiologically active material is at least one selected from the group consisting of compounds, extracts, proteins, peptides, nucleic acids, and antibodies. (a) mixing mastic gum or gamma oryzanol with lipid and natural or synthetic oil
(b) as an oil phase, by melting the mixture prepared in step (a) and mixing mastic gum or gamma oryzanol with lipids and natural or synthetic oils and fats in the mixture, mastic gum or gamma oryzanol, lipids, and natural oils or fats or Step of preparing a mixture containing synthetic oil
(c) preparing a lipid nanoparticle composition by cooling a mixture containing mastic gum or gamma oryzanol, lipid, and natural or synthetic oil prepared in step (b)
(d) preparing the award; and
(e) adding and homogenizing the lipid nanoparticle composition prepared in step (c) to the aqueous phase prepared in step (d), or hydrating the aqueous phase prepared in step (d) and then freeze-drying,
A method for producing lipid nanoparticles and a composition according to claim 1 comprising a. The method according to claim 11, wherein the amount of mastic gum or gamma oryzanol added in step (a) is 0.05 to 50% by weight based on the weight of the lipid. The method of claim 11, wherein the oil is 1.4 to 2.5 parts by weight based on 1 part by weight of the lipid.

Images