Albumin nanocomplex containing phytochemicals and composition containing the same for improving muscle diseases

Abstract

The present invention relates to a phytochemical-containing albumin nanocomplex and a composition containing the same for improving muscle diseases. More specifically, the present invention relates to a phytochemical-containing albumin nanocomplex that has the effect of suppressing muscle loss caused by oxidative stress or inflammatory responses and promoting the differentiation of myoblasts into muscle cells. The phytochemical-containing albumin nanocomplex effectively delivers phytochemicals to immune cells that induce reactive oxygen species and inflammatory responses, thereby regulating the muscle loss signaling mechanism induced by reactive oxygen species and inflammatory responses, suppressing muscle cell atrophy, and promoting differentiation. This can improve muscle loss and prevent and treat muscle atrophy. Therefore, a composition containing the phytochemical-containing albumin nanocomplex of the present invention can be provided as a composition for improving muscle diseases.

Description

[Technical Field]

[0001] The present invention relates to an albumin nanocomposite containing a phytochemical and a composition containing the same for improving muscle diseases. More specifically, the present invention relates to an albumin nanocomposite containing a phytochemical that prevents or improves muscle diseases by suppressing muscle loss caused by oxidative stress or inflammatory responses and promoting the differentiation of myoblasts into muscle cells, and a composition containing the same for improving muscle diseases. [Background technology]

[0002] In recent years, the elderly population has been rapidly increasing worldwide, and according to the United Nations, the population aged 60 and over is predicted to exceed 2 billion by 2050. With this rapid increase in the elderly population aged 65 and over, the quality of life of the elderly in an aging society has become an important issue, and there is a growing demand for extending healthy life expectancy. Skeletal muscle is the largest organ in the human body, accounting for 40-50% of total body weight. It is involved in a variety of functions, including maintaining physique, physical activities such as walking, energy homeostasis, heat production, and glucose and amino acid metabolism. Sarcopenia, which is accompanied by a decrease in skeletal muscle mass and muscle function, is one of the most common symptoms of aging. With the WHO assigning a disease code to sarcopenia in 2017, experts predict that food and pharmaceutical companies will develop numerous products to prevent and treat the condition. This muscle loss manifests itself through an imbalance between muscle protein production and breakdown, increased muscle cell death, and decreased muscle regeneration. Muscle loss and muscle weakness are among the major physical changes associated with aging and are known to progress gradually from the age of 40. This muscle loss, accompanied by a decline in overall physical function, reduces an individual's quality of life and increases the mortality rate of elderly people with low muscle mass. It has been reported that the mortality rate of elderly people with sarcopenia is significantly higher than that of elderly people with the same disease. Possible causes of sarcopenia include hormone deficiencies, such as growth hormone and sex hormones, an imbalance between muscle protein synthesis and breakdown, inactivity, obesity, increased inflammatory cytokines, decreased mitochondrial function, and insulin resistance. In particular, sarcopenia is reported to be caused by an increase in obesity and visceral fat with aging, which in turn increases inflammatory cytokines. Currently, hormone therapy is the most commonly used treatment for muscle loss, but it is not very effective and side effects are being reported one after another. Many pharmaceutical companies have developed inhibitors that suppress or block myostatin and have conducted numerous clinical trials, but all have failed in clinical trials, and new alternatives are desperately needed. Since sarcopenia is caused by various factors, it has been reported that it is difficult to control sarcopenia by controlling only one pathway. Therefore, in order to increase the success rate in clinical trials, it is necessary to develop a technology that can control multiple pathways of muscle weakness to alleviate and treat sarcopenia. To develop such new materials, Korean Patent No. 10-2216599 discloses the extraction of active ingredients from Japanese river snail and green tea as ingredients for a health functional food for improving sarcopenia, and Korean Patent No. 10-2606636 discloses a technology for providing a composition for improving sarcopenia, confirming that a Bacillus verrezensis strain or its culture medium has the effect of preventing or improving sarcopenia. However, none of the above documents discloses a composition that can selectively deliver phytochemicals to tissues where muscle loss has been induced, thereby improving the muscle disease improving effect.

[0003] technical challenges

[0004] An object of the present invention is to provide albumin nanocomplexes containing phytochemicals. Another object of the present invention is to provide a pharmaceutical composition for preventing or treating muscle diseases, comprising the albumin nanocomplex. It is yet another object of the present invention to provide a food composition for preventing or improving muscle diseases, which comprises the albumin nanocomplex.

[0005] The present invention provides a nanocomposite comprising albumin bound to an azide group (N3) or a cyclooctyne group and a delivery substance bound to an azide group (N3) or a cyclooctyne group, which is obtained by a click chemistry reaction, and a phytochemical supported on the surface of the albumin, wherein the delivery substance contains a glucosyl group, and when the albumin is bound to the azide group, the delivery substance is bound to the cyclooctyne group, and when the albumin is bound to the cyclooctyne group, the delivery substance is bound to the azide group. In the present invention, the phytochemical may be one or more selected from the group consisting of phenolic compounds and triterpenes. In the present invention, the phenolic compounds include apigenin, citrusinol, flavone, flavonol, flavanone, flavanol, isoflavone, anthocyanins, stilbenoid, caffeic acid, p-coumaric acid, and the like. The active ingredient may be one or more selected from the group consisting of catechin, chrysin, tectochrysin, primetin, acacetin, luteolin, tangeritin, quercetin, kaempferol, genistein, daidzein, naringenin, hesperetin, cyanidin, catechin, curcumin, epigallocatechin gallate, and resveratrol. In the present invention, the triterpene may be at least one selected from the group consisting of ursolic acid, soyasapogenol A, soyasapogenol B, ursane, oleanane, lupane, saponin, sterol, oleanolic acid, lupeol, corosolic acid, maslinic acid, betulinic acid, ginsenoside, β-sitosterol, campesterol, stigmasterol, lanosterol, and germanicol. In the present invention, the number of azide groups or cyclooctyne groups introduced into the albumin may be 1 to 14. In the present invention, the nanocomposite may contain 4 to 8 glucosyl groups. In the present invention, the number of the phytochemicals supported can be 1 to 20. In the present invention, an azide group or a cyclooctyne group may be bound to the 6-carbon of the glucosyl group. In the present invention, the delivery substance further comprises a radioisotope, and the radioisotope is 3 H, 11 C. 18 F, 14 Cl, 32 P, 35 S, 36 Cl, 45 Ca, 51 Cr, 57 Co, 58 Co, 59 F, 64 Cu, 67 Ga, 68 Ga, 89 Zr, 90 Y, 99 Mo, 99m Tc,111 In, 131 I, 125 I, 124 I, 123 I, 186 Re, 188 Re, 225 Ac, 212 Pb, 117m Sn and 177 Lu. In the present invention, the radioisotope is labeled with a chelating agent, and the chelating agent is not particularly limited to NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid), DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid), DFO (3-[6,17-dihyroxy-7,10,18,21-tetraoxo-27-[N-acetylhydroxylamino]-6,11,17,22-tetraazaheptaeicosane]thiourea), DTPA (diethylenetriaminepentaacetic acid), N2S2 (diaminedithiol), p-SCN-Bn-NOTA (2-(4'-isothiocyanatobenzyl)-1,4,7-triazacyclononane-1,4,7-triacetic acid), NODAGA (1,4,7-triazacyclononane,1-glutaric acid-4,7-acetic acid), p-SCN-Bn-DOTA (2-(4'-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane1,4,7,10-tetraacetic acid), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), p-SCN-Bn-DTPA (2-(4-isothiocyanatobenzyl)-diethylenetriaminepentaacetic acid), p-SCN-Bn-DFO (1-(4-isothiocyanatophenyl)-3-[6,17-dihyroxy-7,10,18,21-tetraoxo-27-[N-acetylhydroxylamino]-6,11,17,22-tetraazaheptaeicosane]thiourea) and HYNIC (hydrazinonicotinic acid). In the present invention, the delivery substance further comprises a fluorescent substance, and the fluorescent substance may be one or more selected from the group consisting of FNR (Ferrodoxin NADP(+) reductase), cyanine fluorescent substances, TAMRA (tetramethylrhodamine-5-maleimide), Flamma (registered trademark) fluorescent substance, and ICG (indocyanine green). In the present invention, the nanocomplex may selectively target macrophages in which GLUT (Glucose Transporter) is overexpressed, and deliver phytochemicals thereto.

[0006] The present invention also provides a pharmaceutical composition for preventing or treating muscle diseases, comprising the albumin nanocomplex. In the present invention, the muscle disease may be caused by an anti-cancer drug or an inflammatory reaction. In the present invention, the anticancer drug may be one or more selected from the group consisting of etoposide, adriamycin, vincristine, cyclophoshamide, ifosphamide, cisplatin, vinorelbine, paclitaxel, docetaxel, gemcitabine, and pemetrexed. In the present invention, the muscle disease can be selected from the group consisting of sarcopenia and muscle atrophy. In the present invention, the pharmaceutical composition may inhibit muscle loss and promote the proliferation of myoblasts and their differentiation into muscle cells. The present invention also provides a food composition for preventing or improving muscle diseases, comprising the albumin nanocomplex. Effect of the invention

[0007] The albumin nanocomplex containing phytochemicals according to the present invention can effectively deliver phytochemical substances by targeting immune cells that induce reactive oxygen species and inflammatory responses, thereby regulating the signaling mechanisms of muscle loss induced by oxidative stress, inflammatory responses, or anticancer drugs, suppressing muscle cell atrophy, and promoting differentiation, thereby improving muscle loss and preventing and treating muscle atrophy. Therefore, the albumin nanocomplex containing phytochemicals according to the present invention can be provided as an excellent composition for improving muscle diseases. [Brief explanation of the drawings]

[0008] FIG. 1 shows an apigenin-loaded albumin nanocomplex according to one embodiment of the present invention. FIG. 2 shows the results of UV analysis confirming the number of apigenins bound to the surface of glycated albumin according to one embodiment of the present invention. FIG. 3 shows the results of UV analysis to evaluate the degree of drug release over time and confirm the stability of apigenin-loaded glycated albumin according to one embodiment of the present invention. FIG. 4 shows the results of examining the hydrogen peroxide scavenging ability to confirm the ROS scavenging effect of apigenin-glycated albumin according to one embodiment of the present invention. FIG. 5 shows the results of examining the oxygen radical scavenging ability to confirm the ROS scavenging effect of apigenin-glycated albumin according to one embodiment of the present invention. FIG. 6 shows the results of measuring the intracellular fluorescence expression levels of glycated albumin (Alb-DBCO-Glc) and non-glycated albumin (Alb-DBCO) conjugated with FNR648 fluorescence to determine whether glucose-glycated albumin targets M1 macrophages, according to one embodiment of the present invention. FIG. 7 shows the results of examining (a) cell morphology and (b) intracellular / extracellular ROS changes after inducing polarization of a raw cell line from the M0 state to the M1 state to confirm that glycated albumin is selectively taken up by M1 macrophages according to one embodiment of the present invention. After inducing polarization of a raw cell line from the M0 state to the M1 state, glycated albumin (Glc-Abb), non-glycated albumin (DBCO-Alb), apigenin alone, apigenin-bound non-glycated albumin (AA), and apigenin-bound glycated albumin (GA) were transfected with FNR648 fluorescence and treated. FIG. 8 shows the results of immunofluorescence staining confirming the cytotoxicity of apigenin-glycated albumin according to one embodiment of the present invention, where FIG. 8a shows the results showing the fluorescent expression of nuclei stained with DAPI, and FIG. 8b is a graph showing the number of nuclei stained with DAPI. FIG. 9 shows the results of immunofluorescence staining confirming the muscle differentiation-promoting effect of apigenin-glycated albumin according to one embodiment of the present invention. FIG. 9a shows the immunofluorescence results of MHC-positive myotubes visualized in green, and DAPI-labeled nuclei shown in blue. FIG. 9b is a graph showing the differentiation level of mouse myoblasts into myotubes based on the number of total nuclei. FIG. 10 shows the results of immunofluorescence staining confirming the muscle atrophy-improving effect of apigenin-glycated albumin according to one embodiment of the present invention. FIG. 10a shows the results of fluorescence expression analysis confirming the level of differentiation into myotubes after treatment with apigenin alone or apigenin-glycated albumin in cells in which muscle atrophy was induced by a cancer cell culture medium. FIG. 10b is a graph showing the level of differentiation into myotubes. FIG. 11 shows the results of Western blot analysis confirming the effect of apigenin-glycated albumin on the expression of muscle production and degradation-related genes according to one embodiment of the present invention. FIG. 12 shows the results of confirming the effect of apigenin-glycated albumin on improving muscle atrophy that may be caused by the administration of an anticancer drug, according to one embodiment of the present invention. FIG. 12a shows the results of fluorescence expression analysis confirming the level of differentiation into myotubes after treating cisplatin-treated mouse muscle cells with apigenin alone or apigenin-glycated albumin, and FIG. 12b is a graph showing the level of differentiation into myotubes. FIG. 13 shows the results of Western blot analysis using proteins extracted from cells in each experimental group of FIG. 12, according to one embodiment of the present invention, to confirm the effect of apigenin-glycated albumin on myogenin expression, which induces muscle formation. FIG. 14 shows the results of confirming the possibility of injecting an albumin nanocomplex with GLUT targeting ability according to one embodiment of the present invention. The results are fluorescence analysis results obtained by directly injecting the albumin nanocomplex into an animal model and observing the amount of albumin nanoplatform remaining at the injection site over time. Figure 15 shows sequential SPECT and PET images of mice intramuscularly injected with apigenin (125I-Api), a complex of apigenin and albumin (125I-Api / 64Cu-Alb), and a complex of apigenin and glycated albumin (125I-Api / 64Cu-Glc-Alb). FIG. 16a shows the results of fluorescence expression analysis confirming the level of differentiation into myotubes after treatment with ursolic acid alone or ursolic acid-glycated albumin in cells in which muscle atrophy was induced by cancer cell culture medium, and FIG. 16b is a graph showing the level of differentiation into myotubes. Figure 17a shows the results of fluorescence expression analysis confirming the level of differentiation into myotubes in cells in which muscle atrophy was induced by cancer cell culture medium after treatment with soyasapogenol B alone or soyasapogenol B-glycated albumin, and Figure 17b is a graph showing the level of differentiation into myotubes. Figure 18a shows the results of a fluorescence expression analysis confirming the level of differentiation into myotubes after treatment with citrusinol alone or citrusinol-glycated albumin in cells in which muscle atrophy was induced by a cancer cell culture medium, and Figure 18b is a graph showing the level of differentiation into myotubes.

[0009] Specific embodiments of the present invention will be described in more detail below. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by a skilled artisan in the art to which the present invention belongs. Generally, the nomenclature used herein is that which is well known and commonly used in the art. The present invention relates to an albumin nanocomplex containing a phytochemical and a composition for improving muscle diseases containing the same. Phytochemicals are a compound word made up of "phyto" (plant-based) and "chemical" (chemical), and refer to plant-derived chemicals that have physiological activities that are beneficial to health. In the present invention, by loading phytocalcines onto the albumin nanoplatform, it is possible to specifically regulate oxidative stress and inflammatory responses induced by anticancer drugs or immune cells, and to demonstrate the effect of improving muscle diseases associated with oxidative stress and inflammatory responses. In the present invention, the phytochemical may be a phenolic compound or a triterpene. Preferred phenolic compounds that can be used in the present invention include flavonoids, stilbenoids, caffeic acid, p-coumaric acid, curcumin, and resveratrol. Flavonoids are a type of secondary metabolite of plants and fungi, and generally have a 15-carbon skeleton consisting of two phenyl rings and a heterocyclic ring. More than 5,000 types of natural flavonoids are known, and research utilizing their diverse activities is ongoing. Examples of the flavonoids include apigenin, citrusinol, flavone, flavonol, flavanone, flavanol, isoflavone, anthocyanins, chrysin, tectochrysin, primetin, acacetin, luteolin, tangeritin, quercetin, kaempferol, genistein, daidzein, naringenin, hesperetin, cyanidin, catechin, and epigallocatechin gallate. gallate), and the like can be used, with apigenin and citrusinol being preferred. Apigenin is a flavonoid, a yellow pigment found in various fruits and vegetables, and is known to be abundant in parsley, chamomile, celery, and carrots. Apigenin has been used for centuries in the form of chamomile tea or applied directly to the skin, but in recent years it has been revealed to be a molecule that can regulate various intracellular mechanisms, and in particular has been reported to suppress inflammation and oxidative stress and promote carbohydrate metabolism. In the present invention, as shown in Figure 1, apigenin is incorporated into an albumin-based nanocomposite, which targets activated M1-type inflammatory macrophages present in the periphery of acute inflammation or cancerous sites. By precisely delivering apigenin to the target site, it is possible to provide an albumin nanocomposite and a composition containing the same, which can suppress muscle loss induced by oxidative stress or inflammatory responses and promote muscle cell differentiation, thereby improving muscle diseases. Citrusinol is a flavonoid compound found in plants such as Citrus unshiu and Philodium pulchellum. According to the present invention, by incorporating citrusinol into an albumin nanoplatform, an albumin nanocomposite capable of improving muscle diseases can be provided. Triterpenes are a group of terpenes, the main components of plant oils, that have a carbon skeleton of 30 carbon atoms, and most have a tetracyclic or pentacyclic structure. Triterpenes are known to have anti-inflammatory, antioxidant, and anti-cancer activities. In the present invention, the triterpene may be ursolic acid, soyasapogenol A, soyasapogenol B, ursane, oleanane, lupane, saponin, sterol, oleanolic acid, lupeol, corosolic acid, maslinic acid, betulinic acid, ginsenoside, β-sitosterol, campesterol, stigmasterol, lanosterol, germanicol, or the like, with ursolic acid and soyasapogenol B being preferred. Phytochemicals delivered by the albumin nanocomplex of the present invention can specifically regulate oxidative stress and inflammatory responses induced by anticancer drugs or immune cells, thereby improving muscle diseases associated with oxidative stress and inflammatory responses. In particular, the albumin nanocomplex of the present invention not only overcomes the poor water solubility and delivery capacity of natural active ingredients such as apigenin, but also effectively scavenges intracellular reactive oxygen species (ROS), demonstrating superior efficacy compared to treatment with phytochemicals alone. This effect was demonstrated through experiments at the immune cell level. Furthermore, treatment of muscle cells with the albumin nanocomplex increased the expression of muscle cell differentiation-related proteins, promoting muscle cell differentiation and demonstrating the associated effects of alleviating inflammation and improving muscle function.

[0010] One aspect of the present invention provides an albumin nanocomplex obtained by a click chemistry reaction between albumin having an azide group (N3) or a cyclooctyne group bound thereto and a delivery substance having an azide group (N3) or a cyclooctyne group bound thereto, wherein a phytochemical is supported on the surface of the albumin, the delivery substance contains a glucosyl group, and when the albumin is bound to the azide group, the delivery substance is bound to the cyclooctyne group, and when the albumin is bound to the cyclooctyne group, the delivery substance is bound to the azide group. In the present invention, albumin refers to a protein that is one of the proteins that constitute the basic substances of cells, is present in large amounts in blood, and is produced in the liver. Albumin has the lowest molecular weight among simple proteins found in nature. Serum albumin in blood maintains and restores plasma volume, prevents shock caused by excessive bleeding, and is used in surgery and burn treatment. It is also known to have the same oxygen delivery capacity as hemoglobin. The albumin may include any albumin that can be formulated, but is preferably, but not limited to, albumin derived from human plasma or recombinant human serum albumin produced by genetic engineering. Genetic information for the albumin of the present invention can be obtained from known databases such as NCBI GenBank. In the present invention, the number of amino groups (-NH2) exposed on the surface of albumin can be 15 to 30. In one specific embodiment of the present invention, to prepare a nanocomposite based on biocompatible albumin that can deliver a delivery substance such as a drug or fluorescent substance to a target tissue in the body and exhibit the effect of treating a disease, a representative human serum albumin (HSA) was used, and one of the functional groups for click chemistry, cyclooctyne or azide group (N3), was introduced onto the surface of the human serum albumin under reaction conditions that minimized denaturation of HSA. In the present invention, azide (N3) is a reactive group consisting of three nitrogen atoms and is highly reactive. In particular, it is known to act as an electron donor in the 1,3-dipolar cycloaddition reaction, which is a type of Cu-free click chemistry, and to play a role in forming a triaza-5-membered ring. The cyclooctyne group is an eight-membered aliphatic ring containing a triple bond under ring strain. It is known to act as an electron acceptor, particularly in the 1,3-dipolar cycloaddition reaction, a type of Cu-free click chemistry, forming a triaza-5-membered ring. The structural feature of cyclooctyne, i.e., the triple bond structure under ring strain, enables click chemistry without the need for a Cu(I) catalyst. The cyclooctyne group is 4-cyclooctyn-1-yl( JPEG2026507017000002.jpg2720), 3-cyclooctyn-1-yl( JPEG2026507017000003.jpg2426), 2-cyclooctyn-1-yl( JPEG2026507017000004.jpg2122), Monofluorinated cyclooctyne(MOFO)( JPEG2026507017000005.jpg2025), Difluororinated cyclooctyne(DIFO)( JPEG2026507017000006.jpg2428), Dimethoxyazacyclooctyne(DIMAC)( JPEG2026507017000007.jpg2735), Dibenzocyclooctyne(DIBO)( JPEG2026507017000008.jpg2542), Azadibenzocyclooctyne(ADIBO)( JPEG2026507017000009.jpg2944) and biarylazacyclooctynone (BARAC) ( JPEG2026507017000010.jpg2843), but is not necessarily limited to this. In the present invention, the number of click reaction functional groups (azide groups or cyclooctyne groups) introduced onto the albumin surface is preferably 1 to 14, and more preferably 9 to 12. In this case, various delivery substances can be injected into the human body in excess, increasing the possibility of uptake into the target site. On the other hand, if the number of click reaction functional groups exceeds the above range, the substance will be immediately taken up by the liver upon injection into the body, which may limit uptake into other target disease sites. The number of click reaction functional groups introduced onto the albumin surface can be adjusted depending on the reaction ratio of albumin to azide-NHS or cyclooctyne-NHS. The solution of albumin having an azide group (N3) bound thereto or an albumin solution having a cyclooctyne group bound thereto can be obtained by (a) dissolving albumin in phosphate-buffered saline (PBS), (b) dissolving azide-NHS or cyclooctyne-NHS in DMSO, and (c) mixing the resulting solutions and then reacting them at 20 to 37°C for 30 minutes to 1 hour. In step (a), the phosphate buffer solution (PBS) may have a pH of 6.8 to 7.6, preferably 7.0 to 7.4. In step (b), the amount of DMSO used to prepare the azide-NHS solution or the cyclooctyne-NHS solution may be 2% (v / v) or less of the total reaction solution. In step (c), the mixing molar ratio of albumin to azide-NHS or cyclooctyne-NHS can be 1:1 to 1:25. In step (c), when the albumin solution and the azide-NHS solution are mixed, the functional group bound to the albumin may be an azide group, and in step (c), when the albumin solution and the cyclooctyne-NHS solution are mixed, the functional group bound to the albumin may be a cyclooctyne group. In one example of the present invention, a human serum albumin (HSA) solution and an ADIBO-NHS solution were mixed and reacted at 37°C for 30 minutes to prepare HSA-ADIBO. The number of click functional groups (azide or cyclooctyne groups) introduced onto the albumin surface is preferably 1 to 14, and more preferably 10 to 12. This allows for the injection of various delivery substances into the human body in excess, increasing the likelihood of uptake at the target site. In contrast, if the number of click functional groups exceeds the above range, the substance will be immediately taken up by the liver upon injection, potentially limiting uptake at other target disease sites. The number of click functional groups introduced onto the albumin surface can be adjusted by the reaction ratio of albumin to azide-NHS or cyclooctyne-NHS. The albumin nanocomplex of the present invention can be obtained by a click chemistry reaction by mixing a solution containing albumin to which the above-mentioned azide group (N3) or cyclooctyne group is bound with a solution containing a delivery substance to which the azide group (N3) or cyclooctyne group is bound, and the click chemistry reaction can be a Cu-free click chemistry reaction.

[0011] In one embodiment of the present invention, the azide group used as the click chemistry functional group is an electron donor, and the cyclooctyne group is an electron acceptor. Therefore, when the functional group bound to albumin is an azide group, the functional group bound to the delivery substance is preferably a cyclooctyne group, and when the functional group bound to albumin is a cyclooctyne group, the functional group bound to the delivery substance is preferably an azide group. In the present invention, the albumin nanocomplex can contain a phytochemical on the surface of the albumin. The phytochemicals supported by the nanocomposite include the aforementioned phytochemicals and their derivatives. For example, apigenin derivatives refer to compounds in which some of the oxygen or hydrogen atoms in apigenin have been replaced with other elements or substituents. The phytochemicals are supported on the surface of albumin, and it is believed that there are pockets on the albumin surface that can specifically support the phytochemicals, allowing the phytochemicals to be stably supported and form a nanocomposite. The nanocomplex of the present invention can precisely deliver phytochemicals supported on the albumin surface to target sites, thereby suppressing muscle loss induced by oxidative stress, inflammatory responses, or anticancer drug administration, and improving muscle diseases by promoting muscle cell differentiation. In particular, the apigenin-loaded albumin nanocomplex can exhibit a synergistic effect in improving muscle diseases when used in the form of glycated albumin, which will be described later. The nanocomplex may contain 1 to 20 phytochemicals on the surface of albumin, more preferably 2 to 10 phytochemicals, and even more preferably 4 to 8 phytochemicals. The albumin nanocomposite of the present invention has a large number of click reaction functional groups, allowing various delivery substances to be bound to a large number of reaction chambers. In the present invention, the delivery substance refers to a substance that is bound to albumin and delivered into the body, and in the present invention, contains a glucosyl group, the structure of which is shown in Formula 1 below. [Formula 1] JPEG2026507017000011.jpg132140 The albumin nanocomplex of the present invention uses glucose as a targeting ligand to deliver phytochemicals by targeting GLUT (glucose transporter) overexpressed in M1-type macrophages. In the present invention, the albumin nanocomplex may contain 4 to 8 glucosyl groups, more preferably 5 to 7. In this case, the nanocomplex injected into the body remains in the injection site or blood for a long time, increasing the target site targeting potential. However, if the number of glucosyl groups exceeds 8, the nanocomplex is immediately taken up by the liver upon injection, which may limit targeting to the target disease site. In the present invention, the glucosyl group, which is a delivery substance, can be bound by a click chemistry reaction between an azide group or cyclooctyne group bound to albumin and an azide group or cyclooctyne group bound to the glucosyl group. In this case, GLUT overexpressed on the surface of M1 cells recognizes the OH groups at positions 1, 3, and 5 of glucose to obtain glucose for use as an energy source. In the present invention, to more effectively target M1 macrophages, albumin can be bound to the 1, 2, and 6 carbon positions of glucose, with position 6 being the most preferred. For example, a glucosyl group with an azide attached to the 1, 2, or 6 carbon position corresponds to the chemical structures shown below in Formula 2, Formula 3, and Formula 4, respectively. [Formula 2] JPEG2026507017000012.jpg93162[Formula 3] JPEG2026507017000013.jpg139147[Formula 4] JPEG2026507017000014.jpg125136 The nanocomplex in which albumin is bound to the 6th position of glucose can exhibit GLUT targeting ability that is three times and two times higher than that of the nanocomplexes in which albumin is bound to the 1st and 2nd carbons of glucose, respectively. In the present invention, the nanocomplex may further contain one or more substances as delivery substances. In this case, multiple substances can be simultaneously subjected to a click chemistry reaction with albumin having an azide group or a cyclooctyne group bound thereto, or multiple substances can be subjected to a click chemistry reaction sequentially. In the present invention, the delivery substance further includes a radioisotope, which refers to an element with the same atomic number but a different atomic mass, and an isotope having radioactivity is called a radioisotope. Such radioisotopes can be used as important labeling substances in diagnosing diseases and confirming pharmacokinetics by utilizing their property of emitting gamma rays or other subatomic particles to decay radioactivity. Radioisotopes that can be used as labeling substances in the present invention can be any radioisotope known in the art without limitation. 3 H, 11 C. 18 F, 14 Cl, 32 P, 35 S, 36Cl, 45 Ca, 51 Cr, 57 Co, 58 Co, 59 F, 64 Cu, 67 Ga, 68 Ga, 89 Zr, 90 Y, 99 Mo, 99m Tc, 111 In, 131 I, 125 I, 124 I, 123 I, 186 Re, 188 Re, 225 Ac, 212 Pb, 117m Sn, and 177 Lu. Preferably, 11 C. 18 F, 64 Cu, 67 Ga, 68 Ga, 89 Zr, 99m Tc, 111 In and 123 I, etc., but is not necessarily limited to this. In the present invention, the radioisotope is labeled with a chelating agent, which serves to link the radioisotope to albumin. Examples of the chelating agent include NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid), DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid), DFO (3-[6,17-dihyroxy-7,10,18,21-tetraoxo-27-[N-acetylhydroxylamino]-6,11,17,22-tetraazaheptaeicosane]thiourea), DTPA (diethylenetriaminepentaacetic acid), N2S2 (diaminedithiol), p-SCN-Bn-NOTA (2-(4'-isothiocyanatobenzyl)-1,4,7-triazacyclononane-1,4,7-triacetic acid), and the like. acid), NODAGA (1,4,7-triazacyclononane,1-glutaric acid-4,7-acetic acid), p-SCN-Bn-DOTA (2-(4'-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), p-SCN-Bn-DTPA (2-(4-isothiocyanatobenzyl)-diethylenetriaminepentaacetic acid), p-SCN-Bn-DFO The compound may be one or more selected from the group consisting of (1-(4-isothiocyanatophenyl)-3-[6,17-dihyroxy-7,10,18,21-tetraoxo-27-[N-acetylhydroxylamino]-6,11,17,22-tetraazaheptaeicosane]thiourea), and HYNIC (hydrazinonicotinic acid), but is not necessarily limited thereto. In the present invention, the delivery substance may further include a fluorescent substance. A fluorescent substance is a substance that emits a specific wavelength of visible light at a specific wavelength, and in the present invention includes all fluorescent substances that can be bound to albumin nanocomplexes and used to identify the location of the nanocomplexes. Specifically, fluorescent substances that can be used as labeling substances in the present invention can be any fluorescent substance known in the art, and include, but are not limited to, rhodamine-based substances such as rhodamine, TAMRA, etc.; fluorescein-based substances such as fluorescein, FITC (fluorescein isothiocyanate), and FAM (fluorescein amidite); bodipy-based substances (bodipy, boron-dipyrromethene); Alexa Fluor-based substances; and cyanine-based substances such as Cy3, Cy5, Cy7, and indocyanine green.

[0012] Another aspect of the present invention provides a pharmaceutical composition for preventing or treating a muscle disease, comprising an albumin nanocomplex containing a phytochemical. In the present invention, the muscle disease may be a muscle disease caused by an anticancer drug or an inflammatory reaction. Specifically, the albumin nanocomplex containing the phytochemical of the present invention can suppress muscle loss by regulating the signaling mechanisms that induce muscle weakening due to oxidative stress or inflammatory responses induced by anticancer drug administration and immune cells, and can promote muscle cell differentiation to induce an increase in muscle mass. According to one embodiment of the present invention, the apigenin-loaded glycated albumin nanocomplex selectively targeted inflammatory M1-type macrophages in mouse muscle cells, inhibiting muscle cell loss due to inflammatory responses and promoting myoblast differentiation. Additionally, it also inhibited muscle cell loss and atrophy induced by anticancer drug administration and promoted muscle cell differentiation, thereby improving muscle loss and muscle atrophy. In particular, the cell group administered with the apigenin-loaded glycated albumin nanocomplex showed a significant reduction in ROS compared to the cell group administered with apigenin alone, demonstrating the effect of suppressing the differentiation of macrophages into inflammatory types. Furthermore, in muscle cells in which muscle atrophy was induced by cancer cell culture medium or anticancer drugs, the cell group administered with the apigenin-loaded glycated albumin nanocomplex showed a significantly greater improvement effect than the group administered with apigenin alone, demonstrating the synergistic effect of the apigenin and glycated albumin nanocomplex of the present invention. Furthermore, it has been confirmed that the apigenin-containing albumin nanocomplex suppresses the expression of muscle cell degrading genes induced by immune cells and the administration of anticancer drugs, and increases the expression of genes that induce the differentiation of myoblasts into muscle cells. This indicates that the pharmaceutical composition containing the apigenin-containing albumin nanocomplex of the present invention can prevent or treat muscle diseases by suppressing muscle loss and promoting the proliferation and differentiation of myoblasts into muscle cells. In the present invention, the anticancer drug may be one or more selected from the group consisting of etoposide, adriamycin, vincristine, cyclophoshamide, ifosphamide, cisplatin, vinorelbine, paclitaxel, docetaxel, gemcitabine, and pemetrexed.

[0013] In another example of the present invention, ursolic acid, soyasapogenol B, and citrusinol were formulated as an albumin nanocomplex, similar to apigenin, and it was confirmed that this could inhibit muscle loss and promote myoblast proliferation and differentiation into muscle cells. In particular, it was confirmed that these substances could exert a more excellent synergistic effect than when used alone. In the present invention, the muscle disease can be selected from the group consisting of sarcopenia and muscle atrophy. In the present invention, the term "sarcopenia or muscle atrophy" is interpreted to include not only the dictionary meaning of "sarcopenia" or "muscular atrophy," but also muscle diseases that cause or result from muscle loss or muscle atrophy, such as sarcopenic obesity, myopathy, muscular dystrophy, muscle injury, myasthenia, myoneural conductive disease, nerve injury, amyotrophic lateral sclerosis (ALS), atony, myotonia, cachexia, etc. For example, the term "muscular atrophy" may include diabetic amyotrophy, spinal muscular atrophy, etc. The term "muscular dystrophy" may include Duchenne muscular dystrophy, Becker muscular dystrophy, limb girdle muscular dystrophy, facioscapulohumeral muscular dystrophy, oculopharyngeal muscular dystrophy, myotonic dystrophy, etc. Furthermore, the myopathy may include inflammatory myopathy such as polymyositis and dermatomyositis, endocrine myopathy, toxic myopathy, metabolic myopathy, mitochondrial myopathy, congenital myopathy, and the like.

[0014] The pharmaceutical compositions may further contain suitable pharmaceutically acceptable carriers, excipients, or diluents according to conventional methods. The pharmaceutically acceptable carriers are those commonly used in pharmaceutical formulations, including, but not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition to the above ingredients, the pharmaceutical composition of the present invention may additionally contain lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, etc. Appropriate pharmaceutically acceptable carriers and formulations can be appropriately formulated according to each ingredient using the methods disclosed in Remington's Pharmaceutical Sciences (19th edition, 1995). The pharmaceutical composition of the present invention can be administered either orally or parenterally. Parenteral administration includes intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, and the like. Oral dosage forms include tablets, pills, hard capsules, soft capsules, liquids, suspensions, emulsions, syrups, and granules. These formulations may contain, in addition to the active ingredient, diluents (e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and / or glycine), lubricants (e.g., silica, talc, stearic acid and its magnesium or calcium salts, and / or polyethylene glycol). The tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidone, and may optionally contain disintegrants or effervescent mixtures such as starch, agar, alginic acid, or its sodium salt, and / or absorbents, colorants, flavorings, and sweeteners. These formulations can be prepared by conventional mixing, granulating, or coating methods. A typical example of a formulation for parenteral administration is an injection formulation, and examples of solvents for the injection formulation include water, Ringer's solution, isotonic saline, and suspensions. Sterile fixed oils can be used as a solvent or suspending medium for the above-mentioned injection preparations, and any non-irritating fixed oils including mono-glycerides and di-glycerides can be used for this purpose.Furthermore, the above-mentioned injection preparations can use fatty acids such as oleic acid. The compositions of the present invention are administered in a pharmaceutically effective amount. In the present invention, a "pharmaceutically effective amount" refers to an amount sufficient to treat a disease based on a reasonable benefit / risk ratio applicable to any medical treatment. The effective dosage can be determined based on factors including the type and severity of the patient's disease, the activity and sensitivity of the drug, the administration time, administration route and excretion rate, the duration of treatment, concurrently used drugs, and other factors well known in the medical field. The compositions of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents. They can be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered in a single dose or multiple doses. Taking all of the above factors into consideration, it is important to administer an amount that achieves maximum efficacy with the minimum dose without causing side effects, which can be easily determined by one of ordinary skill in the art. Specifically, the effective amount of the composition of the present invention may vary depending on the age, sex, and weight of the patient, but is generally 0.001 to 150 mg per kg of body weight, preferably 0.01 to 100 mg, administered daily or every other day, or in 1 to 3 divided doses per day. However, since the amount may increase or decrease depending on the route of administration, sex, weight, age, etc., the above dosage does not limit the scope of the present invention in any way.

[0015] The present invention also provides a food composition for preventing or improving muscle diseases, which contains a nanocomposite containing a phytochemical. The food of the present invention is not limited to human food, but may also include animal food and feed additives. In another embodiment of the present invention, the food composition may further contain one or more additives selected from the group consisting of organic acids, phosphates, antioxidants, lactose casein, dextrin, glucose, sugar, and sorbitol. The organic acids may be, but are not limited to, citric acid, fumaric acid, adipic acid, lactic acid, or malic acid. The phosphates may be, but are not limited to, sodium phosphate, potassium phosphate, acid pyrophosphate, or polyphosphate (polymerized phosphate). The antioxidants may be, but are not limited to, natural antioxidants such as polyphenols, catechins, α-tocopherol, rosemary extract, licorice extract, chitosan, tannic acid, or phytic acid. In yet another embodiment of the present invention, the food composition may contain, in addition to the active ingredients, various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and flavor enhancers (for cheese, chocolate, etc.), pectinic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated beverages, etc. Additionally, the food composition according to one embodiment of the present invention may contain fruit pulp for producing natural fruit juice, fruit juice beverages, and vegetable beverages. According to one embodiment of the present invention, the food composition formulation may be in the form of, but is not limited to, a solid, powder, granule, tablet, capsule, liquid or beverage. Furthermore, the food composition can be used to produce foods such as, but not limited to, confectioneries, sugars, ice cream products, dairy products, meat products, fish products, beans or agar, edible oils and fats, noodles, tea, beverages, special nutritional foods, health supplements, seasonings, ice, ginseng products, kimchi pickles, dried meat, fruits, vegetables, dried fruit or vegetable products, cut products, fruit juice, vegetable juice, mixed juices thereof, chips, noodles, processed livestock foods, processed seafood foods, processed dairy foods, fermented milk foods, bean foods, grain foods, microbially fermented foods, confectioneries and breads, seasonings, processed meat products, acidic beverages, licorice, and herbs. Thus, the albumin nanocomplex containing phytochemicals according to the present invention effectively delivers phytochemicals by targeting immune cells that induce reactive oxygen species and inflammatory responses, thereby regulating the muscle loss signaling mechanism induced by oxidative stress and inflammatory responses, suppressing muscle cell atrophy, and promoting differentiation, thereby improving muscle loss and achieving the effects of preventing and treating muscle atrophy.

[0016] Example The present invention will be described in more detail with reference to the following examples, but these examples are merely illustrative of some experimental methods and configurations for the purpose of illustrating the present invention, and the scope of the present invention is not limited to these examples.

[0017] Production Example 1: Production of apizenin-glycated albumin complex 1-1. Production of glycated albumin complex Glycated albumin having glucosyl groups bound thereto was produced by the following method. Albumin was dissolved in phosphate-buffered saline (PBS, pH 7.4) at a concentration of 40 mg / mL, and the albumin solution was dispensed at 20 mg / 0.5 mL per vial. DBCO-NHS was dissolved in DMSO (10 mg / 50 μL) and added to the albumin solution to prepare a 500 μL solution. Albumin and DBCO-NHS were reacted at a ratio of 1:14 at room temperature for 6 hours, and then purified using an Amicon Ultra-0.5 centrifugal filter tube to obtain DBCO-albumin (DBCO-Alb). For glucose-albumin synthesis, 6-azido-glucose was dissolved in distilled water at a concentration of 3 mg / mL and added to a DBCO-albumin (DBCO-Alb) solution (4 mg / 0.5 mL). The mixture was incubated at room temperature for 1 hour and then purified using a centrifugal filter tube. Next, we used UV-Vis spectroscopy to confirm the absorbance at 280 nm and 309 nm of glucose-DBCO-Alb (Alb-DBCO-Glc). To measure the absorbance of albumin, we diluted each albumin sample to 2 mg / mL and performed MALDI-TOF MS to determine the number of ADIBO and glucose conjugated to albumin (degree of functionalization, or DOF). Then, UV and MALDI-TOF analyses were used to measure the number of DBCO molecules on the albumin surface and the number of glucose molecules introduced through the click reaction. 11 -Alb-FL and Glc 6 -DBCO 11 To prepare -Alb-FL, N3-FNR648 (10 nmol / 1 μL, dissolved in DMSO) was added to DBCO-Alb and Glc-DBCO-Alb (30 nmol / 500 μL), incubated at 4°C for 30 min, and then purified using a centrifugal filter tube. The number of DBCO functional groups introduced into HSA-DBCO was quantified using UV-Vis spectrophotometric methods, and was found to be 12.8 to 13.1. When quantified using MALDI-TOF, it was found to be 11.1 to 11.5. In this case, the number of glucose molecules bound was found to be 5.4 to 7.8. 1-2. Preparation of apigenin-loaded glycated albumin complex An attempt was made to load apigenin onto glycated albumin prepared as in Preparation Example 1-1. Apigenin was mixed with albumin at 0.5, 1, 2, 4, 8, 16, and 32 equivalents, and the mixture was incubated for 0.5 to 2 hours. The mixture was then purified using a filter and analyzed by UV light. As shown in Figure 2, the peak characteristic of apigenin between 350 and 380 nm saturated at 16 equivalents or more, confirming that the number of apigenin molecules bound to albumin was between 8 and 16. Next, the stability of the conjugate was confirmed by evaluating the degree of drug release over time. Immediately after preparation of the apigenin-loaded glycated albumin was set as time 0, and albumin was sampled at 10 minutes, 30 minutes, 1 hour, 1 hour 30 minutes, 3 hours, 6 hours 50 minutes, 17 hours, 24 hours, and 88 hours. The degree of release was confirmed by UV light and is shown in Figure 3. Referring to FIG. 3, approximately 20% of the drug was released after 88 hours, confirming that apigenin was stably loaded onto glycated albumin to form an apigenin-glycated albumin complex. Experimental example 1: Confirmation of reactive oxygen species (ROS) removal effect In order to confirm the ROS scavenging effect of the apigenin-glycated albumin complex prepared in Production Example 1, an evaluation experiment was carried out to evaluate the ability to scavenge hydrogen peroxide and oxygen radicals, and the results are shown in Figures 4 and 5, respectively. In the experiment, apigenin alone (API), albumin bound to apigenin (AA), and apigenin-glycated albumin (GA) were compared and evaluated, and the apigenin concentration was 12.5 μM. Referring to Figure 4, which shows the hydrogen peroxide scavenging effect, the hydrogen peroxide scavenging ability showed the same level in all groups, confirming that the complexation with albumin does not affect the oxidative stress scavenging ability of apigenin. Furthermore, in Figure 5, which confirmed the oxygen radical scavenging ability, a similar level of effective oxygen radical scavenging effect was also confirmed. This confirmed that apigenin-glycated albumin has the ability to remove ROS, and that the pharmacological efficacy of apigenin is maintained even when it forms a complex with albumin. Experimental Example 2: Confirmation of M1 macrophage targeting and inhibitory effect 2-1. Confirmation of the M1 macrophage targeting ability of glycated albumin The glycated albumin of Preparation Example 1-1 was prepared using glucose as a target ligand to selectively target GLUT (Glucose transporter) that is overexpressed in M1-type macrophages. To confirm whether glucose-glycated albumin can selectively target M1 macrophages, we polarized raw cell lines into M0 and M1 states, and then treated the cells with glycated albumin (Alb-DBCO-Glc) and non-glycated albumin (Alb-DBCO) transfected with FNR648 fluorescence. Referring to FIG. 6, which shows the targeting ability of each albumin administered to cells, glycated albumin showed uptake in M1 that was four times higher than that of non-glycated albumin. Specifically, no significant difference in uptake was observed for either of the two types of albumin in M0, and no significant difference was observed in any of the groups. However, a significant increase in the fluorescence expression of glycated albumin was confirmed in M1, confirming that glycated albumin is a platform that is specifically taken up by M1. 2-2. Confirmation of apigenin-glycated albumin suppression of M1 macrophages As in the previous experiment, we confirmed whether apigenin-glycated albumin could selectively suppress M1-type macrophages. Using the same method as in Experimental Example 2-1, Raw cell lines were polarized from the M0 to M1 state, and then treated with glycated albumin (Glc-Abb), non-glycated albumin (DBCO-Alb), apigenin alone, non-glycated albumin bound to apigenin (D / A), and glycated albumin bound to apigenin (G / A) and transfected with FNR648 fluorescence. Fluorescence expression was then confirmed, and changes in cell morphology and intracellular / extracellular ROS were observed. These results are shown in Figures 7a and 7b, respectively. As a result, a reduction in ROS was confirmed in the experimental groups treated with apigenin alone and apigenin-bound albumin (D / A and G / A), and the morphology of the raw cell line was confirmed to be maintained as M0 type. In particular, glycated albumin bound to apigenin (G / A) significantly reduced ROS compared to other experimental groups and showed the effect of suppressing differentiation into M1 type, confirming that apigenin and glycated albumin exert a synergistic effect. Experimental Example 3: Confirmation of the cytotoxicity of apigenin-glycated albumin Immunofluorescence staining was performed to confirm the cytotoxicity of apigenin-glycated albumin. Mouse muscle cells C2C12 were cultured and treated with apigenin alone or apigenin-glycated albumin at equal concentrations of 2.5 and 5 μM. After culturing for 96 hours, the nuclei were stained with DAPI, and fluorescence expression was confirmed as shown in Figure 8a. The number of stained nuclei was counted and shown in the graph in Figure 8b. Referring to FIG. 8b, which shows the number of nuclei in surviving cells, it was confirmed that neither apigenin alone nor apigenin-glycated albumin exhibited cytotoxicity. Experimental Example 4: Confirmation of the muscle cell differentiation promoting effect of apigenin-glycated albumin To confirm the muscle differentiation promoting effect of apigenin-glycated albumin, immunofluorescence staining was performed. Mouse muscle cells C2C12 were cultured, treated with apigenin alone or apigenin-glycated albumin so that apigenin was administered equally at concentrations of 2.5 and 5 μM, and then differentiated into myotubes for 48 hours. Next, the cells were fixed with 4% formaldehyde solution and incubated overnight at 4°C with primary antibodies MHC (myosin heavy chain, Developmental Studies Hybridoma Bank, Iowa City, Iowa, USA) and DAPI (4',6-Diamidino-2-Phenylindole Dihydrochloride, Sigma-Aldrich, MO, USA), followed by secondary antibody attachment at room temperature for 1 hour. Then, as shown in Figure 9a, MHC-positive myotubes were visualized by immunofluorescence in green, and DAPI-labeled nuclei were displayed in blue. The differentiation of mouse myoblasts into myotubes was confirmed based on the total number of nuclei using the formula (Fusion Index = number of nuclei in myotubes / muscle cells), as shown in Figure 9b. 9a and 9b, it was confirmed that the experimental group treated with apigenin-glycated albumin showed increased differentiation into myotubes compared to the experimental group treated with apigenin alone. In particular, the experimental group treated with 2.5 μM apigenin-glycated albumin showed a 15% increase in myotube differentiation effect compared to the experimental group treated with the same concentration of apigenin alone. This confirmed that apigenin-glycated albumin can increase myoblast differentiation and promote muscle formation. Experimental Example 5: Confirmation of the muscle atrophy improving effect of apigenin-glycated albumin To confirm the muscle atrophy-improving effect of apigenin-glycated albumin, immunofluorescence staining was performed in the same manner as in Experimental Example 3. First, mouse muscle cells C2C12 were cultured, and then the culture medium was replaced with cancer cell culture medium (CCM) containing mouse colon cancer cells CT26 (colon carcinoma cells) to induce muscle atrophy. The cells were then treated with 2.5 and 5 μM concentrations of apigenin alone or apigenin-glycated albumin. After 48 hours of culture, the cells were checked for fluorescence expression to confirm the level of differentiation into myotubes. 10a and 10b, which show the level of myotube differentiation of atrophy-induced myoblasts after treatment with apigenin or apigenin-glycated albumin, it was confirmed that the atrophy-induced myoblasts differentiated into myotubes increased in the apigenin-glycated albumin-treated experimental group compared to the apigenin-treated experimental group. In particular, the experimental group treated with 2.5 μM apigenin-glycated albumin showed a 15% increase in myotube differentiation compared to the experimental group treated with the same concentration of apigenin alone, confirming that apigenin-glycated albumin promotes myoblast differentiation and has the effect of improving atrophied muscles. Meanwhile, to confirm the effect of apigenin-glycated albumin on the expression of genes related to muscle production and degradation, proteins were extracted from the cells of each experimental group and subjected to Western blotting to determine the protein expression levels of each gene. The results are shown in Figure 11. 11, MHC expression increased in the experimental groups in which cells that had undergone atrophy due to cancer cell culture medium (CCM) were treated with apigenin alone or apigenin-glycated albumin, demonstrating that CCM improved muscle cell atrophy. Furthermore, the expression of IGF-1R, p-AKT, and p-Foxo3a, which induce muscle formation, was increased in both the apigenin-only and apigenin-glycated albumin-treated cell groups. Furthermore, in the case of MuRF1, which induces muscle atrophy, the expression level of MuRF1, which was significantly increased by CCM treatment, was reduced by apigenin alone and apigenin-glycated albumin treatment. However, the MuRF1 expression level in the apigenin-glycated albumin-treated cell group was significantly reduced compared to the MuRF1 expression level in the apigenin-glycated albumin-treated cell group. This confirmed that apigenin-glycated albumin exhibited a more pronounced synergistic effect than apigenin alone. Experimental Example 6: Confirmation of the muscle cell atrophy suppression effect of anti-cancer drugs We confirmed the ameliorative effect of apigenin-glycated albumin on muscle atrophy that may be caused by anticancer drug administration. First, mouse muscle cells C2C12 were cultured and treated with 40 μM cisplatin, 2.5 μM and 5 μM apigenin alone, or apigenin-glycated albumin. After 48 hours of culture, immunofluorescence staining was performed to confirm the level of differentiation into myotubes, as shown in Figure 12a and Figure 12b. Referring to Figures 12a and 12b, it was confirmed that cisplatin induced atrophy of muscle cells, but that the atrophy of muscle cells caused by cisplatin was suppressed in the cell groups treated with apigenin or apigenin-glycated albumin. In particular, it was confirmed that the atrophy suppression effect was superior in the cell group treated with glycated albumin carrying apigenin at a concentration of 2.5 μM than in the cell group treated with apigenin alone at a concentration of 2.5 μM. In addition, proteins were extracted from the cells of each experimental group and Western blotting was performed to confirm the expression level of myogenin (MyoG), which induces muscle formation, as shown in Figure 13.The results confirmed that the expression of MHC and MyoG, which was reduced by cisplatin treatment, was increased by apigenin treatment.In particular, it was confirmed that the expression level of MyoG, which induces muscle formation, was significantly increased in the cell group treated with glycated albumin loaded with apigenin. This confirmed that apigenin and glycated albumin have a synergistic effect in improving muscle atrophy caused by anticancer drug administration. Experimental Example 7: Confirmation of in-vivo targeting ability of glycated albumin Using the albumin nanocomplex with GLUT targeting ability of the present invention, the possibility of preparing an injectable formulation via a practical administration route was confirmed. Intramuscular injection experiments were conducted to evaluate how long the albumin nanocomplex can remain at the injection site and deliver the drug to the diseased site when directly injected, and the results are shown in FIG. Referring to Figure 14, an albumin platform (Albumin(11)) with 11 ADIBO functional groups introduced therein and an albumin nanocomposite (Glc(6)-Albumin(11) with glucose introduced therein to ensure GLUT targeting ability in the cell experiment) were intramuscularly injected, followed by isotope-based nuclear medicine imaging evaluation at 0 hours, 3 hours, and 24 hours. The results showed that the amount of residual activity at the injection site was very high at all time points. The above experimental results are extremely important in that they not only solve the problem that conventional drugs are rapidly absorbed after intramuscular injection, but do not accumulate sufficiently at the diseased site, resulting in the failure to achieve the expected pharmacological effect, but also make it possible to set a concentration that will exert the optimal pharmacological effect with a smaller amount. Experimental Example 8: Simultaneous distribution evaluation of glycated albumin and apigenin Different isotopes were introduced into albumin and the drug, and after co-injection into mice, the dynamics of the drug and albumin platforms were visualized and analyzed in real time. A labeling method was established for apigenin using the SPECT nuclide I-125 (125I-Api), albumin was labeled with the PET nuclide Cu-64 using click chemistry (64Cu-Alb), and albumin to which glucose had been introduced was also labeled with Cu-64 to confirm the targeting ability due to glycation (64Cu-Glc-Alb). After intramuscular injection of labeled apigenin (125I-Api), apigenin-albumin complex (125I-Api / 64Cu-Alb), and apigenin-glycated albumin complex (125I-Api / 64Cu-Glc-Alb), SPECT and PET images were acquired sequentially and shown in Figure 15. Referring to the image in Figure 15, in mice that received an intramuscular injection of labeled apigenin (125I-Api), it was confirmed that apigenin remained persistently at the injection site, and the image signal was maintained uniformly, confirming that no absorption, metabolism, or excretion occurred. On the other hand, the apigenin-albumin complex (125I-Api / 64Cu-Alb) and the apigenin-glycated albumin complex (125I-Api / 64Cu-Glc-Alb) were confirmed to recirculate in the blood via lymphatic vessels after intramuscular injection and then be excreted through the intestine. This means that the apigenin / albumin complex may be absorbed, metabolized, and excreted, potentially increasing the drug's absorption rate and allowing it to remain at a specific site, thereby demonstrating its efficacy. Furthermore, the same behavior as seen in the PET images was confirmed up to 8 hours after the study, confirming that the compound exists as a complex until it is absorbed and then separated during metabolism and excretion, suggesting the possibility of targeting specific cells for therapy.

[0018] Production Example 2: Production of ursolic acid-glycated albumin complex Ursolic acid was loaded onto the glycated albumin produced by the method of Production Example 1-1. 10 equivalents of ursolic acid based on albumin were mixed with albumin, and the mixture was reacted for 1.5 hours, and then purified using a filter to produce an ursolic acid-glycated albumin complex. Experimental Example 9: Confirmation of the muscle atrophy improving effect of ursolic acid-glycated albumin An experiment was conducted to confirm the effect of ursolic acid-glycated albumin on improving muscle atrophy. First, mouse muscle cells C2C12 were cultured, and then the culture medium was replaced with cancer cell culture medium (CCM) in which mouse colon cancer cells CT26 (colon carcinoma cells) were cultured to induce muscle atrophy. The cells were then treated with 2.5% ursolic acid alone or with ursolic acid-glycated albumin, and after 48 hours of culture, the cells were checked for fluorescence expression and the level of differentiation into myotube cells was confirmed. Referring to Figures 16a and 16b, MHC expression increased in cells that had undergone atrophy due to cancer cell culture medium (CCM) in both the experimental groups treated with ursolic acid alone and ursolic acid-glycated albumin, confirming that CCM improved muscle cell atrophy. In particular, it was confirmed that MHC expression was further increased in the ursolic acid-glycated albumin-treated cell group compared to when ursolic acid was administered alone, confirming that ursolic acid-glycated albumin exhibits a synergistic effect compared to when ursolic acid was administered alone.

[0019] Production Example 3: Production of soyasapogenol B-glycated albumin complex Based on the glycated albumin produced by the method of Preparation Example 1-1, 10 equivalents of soyasapogenol B was mixed with albumin, reacted for 1.5 hours, and then purified using a filter to produce a soyasapogenol B-glycated albumin complex. Experimental Example 10: Confirmation of the muscle atrophy improving effect of soyasapogenol B-glycated albumin An experiment was conducted to confirm the effect of soyasapogenol B-glycated albumin on improving muscle atrophy. First, mouse muscle cells C2C12 were cultured, and then the culture medium was replaced with cancer cell culture medium (CCM) containing mouse colon cancer cells CT26 (colon carcinoma cells) to induce muscle atrophy. The cells were then treated with a 2.5% concentration of soyasapogenol B alone or with soyasapogenol B-glycated albumin, and after 48 hours of culture, the cells were checked for fluorescence expression to determine the level of differentiation into myotube cells. Referring to Figures 17a and 17b, MHC expression increased in cells that had undergone atrophy due to cancer cell culture medium (CCM) in both experimental groups treated with soyasapogenol B alone and soyasapogenol B-glycated albumin, confirming that CCM improved muscle cells that had undergone atrophy. In particular, we confirmed that MHC expression was significantly increased in the cell group treated with soyasapogenol B-glycated albumin compared to when soyasapogenol B was administered alone, demonstrating that soyasapogenol B-glycated albumin exhibits a superior synergistic effect than when soyasapogenol B was administered alone.

[0020] Production Example 4: Production of citrusinol-glycated albumin complex Based on the glycated albumin prepared by the method of Preparation Example 1-1, 10 equivalents of citrusinol were mixed with albumin, reacted for 1.5 hours, and then purified using a filter to prepare a citrusinol-glycated albumin complex. Experimental Example 11: Confirmation of the muscle atrophy improving effect of citrusinol-glycated albumin An experiment was conducted to confirm the effect of citrusinol-glycated albumin on improving muscle atrophy. First, mouse muscle cells C2C12 were cultured, and then the culture medium was replaced with cancer cell culture medium (CCM) containing mouse colon cancer cells CT26 (colon carcinoma cells) to induce muscle atrophy. The cells were then treated with a 2.5% concentration of citrusinol alone or with citrusinol-glycated albumin, and after 48 hours of culture, the cells were checked for fluorescence expression to determine the level of differentiation into myotube cells. Referring to Figures 18a and 18b, MHC expression increased in cells that had undergone atrophy due to cancer cell culture medium (CCM) in both the experimental groups treated with citrusinol alone and citrusinol-glycated albumin, confirming that CCM improved muscle cell atrophy. In particular, MHC expression was further increased in the citrusinol-glycated albumin-treated cell group compared to when citrusinol was administered alone, demonstrating that citrusinol-glycated albumin exhibits a synergistic effect compared to when citrusinol was administered alone. Although some embodiments of the present invention have been described above, the present invention is not limited to the above-described embodiments, and can be implemented with modifications and variations within the scope of the gist of the present invention, and it should be understood that such modified and varied forms also belong to the technical spirit of the present invention.

Claims

1. Azide group (N 3 ) or cyclooctyne group-bound albumin, and azide group (N 3 a nanocomposite obtained by a click chemistry reaction between a delivery substance having a cyclooctyne group or a cyclooctyne group bound thereto and a phytochemical supported on the surface of the albumin, the delivery agent comprises a glucosyl group; An albumin nanocomplex, wherein when the albumin is conjugated with an azide group, the delivery agent is conjugated with a cyclooctyne group, and when the albumin is conjugated with a cyclooctyne group, the delivery agent is conjugated with an azide group.

2. 2. The albumin nanocomplex according to claim 1, wherein the phytochemical is one or more selected from the group consisting of phenolic compounds and triterpenes.

3. The phenolic compounds include apigenin, citrusinol, flavone, flavonol, flavanone, flavanol, isoflavone, anthocyanins, stilbenoid, caffeic acid, p-coumaric acid, and the like. The compound is one or more selected from the group consisting of catechin, tectochrysin, primetin, acacetin, luteolin, tangeritin, quercetin, kaempferol, genistein, daidzein, naringenin, hesperetin, cyanidin, catechin, curcumin, epigallocatechin gallate, and resveratrol. The albumin nanocomplex of claim 2.

4. 3. The albumin nanocomplex according to claim 2, wherein the triterpene is one or more selected from the group consisting of ursolic acid, soyasapogenol A, soyasapogenol B, ursane, oleanane, lupane, saponin, sterol, oleanolic acid, lupeol, corosolic acid, maslinic acid, betulinic acid, ginsenoside, β-sitosterol, campesterol, stigmasterol, lanosterol, and germanicol.

5. The albumin nanocomplex according to claim 1, wherein the number of azide groups or cyclooctyne groups introduced into the albumin is 1 to 14.

6. 2. The albumin nanocomplex of claim 1, wherein the nanocomplex contains 4 to 8 glucosyl groups.

7. The albumin nanocomplex of claim 1, wherein an azide group or a cyclooctyne group is bound to the 6-carbon of the glucosyl group.

8. The albumin nanocomplex according to claim 1, wherein 1 to 20 of the phytochemicals are supported.

9. The delivery agent further comprises a radioisotope, the radioisotope comprising: ３ H, １１ C. １８ F, １４ Cl, ３２ P, ３５ S, ３６ Cl, ４５ Ca, ５１ Cr, ５７ Co, ５８ Co, ５９ F, ６４ Cu, ６７ Ga, ６８ Ga, ８９ Zr, ９０ Y, ９９ Mo, ９９ｍ Tc, １１１ In, １３１ I, １２５ I, １２４ I, １２３ I, １８６ Re, １８８ Re, ２２５ Ac, ２１２ Pb, １１７ｍ Sn, and １７７ The albumin nanocomplex according to claim 1, wherein the albumin nanocomplex is one or more selected from the group consisting of Lu.

10. The radioisotope is labeled with a chelating agent, and the chelating agent is not particularly limited, and may be, for example, NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid), DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid), DFO (3-[6,17-dihyroxy-7,10,18,21-tetraoxo-27-[N-acetylhydroxylamino]-6,11,17,22-tetraazaheptaeicosane]thiourea), DTPA (diethylenetriaminepentaacetic acid), N2S2 (diaminedithiol), p-SCN-Bn-NOTA (2-(4'-isothiocyanatobenzyl)-1,4,7-triazacyclononane-1,4,7-triacetic acid), or NODAGA (1,4,7-triazacyclononane,1-glutaric acid-4,7-acetic acid), p-SCN-Bn-DOTA (2-(4'-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane1,4,7,10-tetraacetic acid), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), p-SCN-Bn-DTPA (2-(4-isothiocyanatobenzyl)-diethylenetriaminepentaacetic acid), p-SCN-Bn-DFO (1-(4-isothiocyanatophenyl)-3-[6,17-dihyroxy-7,10,18,21-tetraoxo-27-[N-acetylhydroxylamino]-6,11,17,22-tetraazaheptaeicosane]thiourea) 10. The albumin nanocomplex according to claim 9, wherein the nanocomplex is one or more selected from the group consisting of HYNIC (hydrazinonicotinic acid).

11. 2. The albumin nanocomplex according to claim 1, wherein the delivery substance further comprises a fluorescent substance, and the fluorescent substance is one or more selected from the group consisting of FNR (Ferrodoxin NADP(+) reductase), cyanine fluorescent substances, TAMRA (tetramethylrhodamine-5-maleimide), Flamma (registered trademark) fluorescent substance, and ICG (indocyanine green).

12. The albumin nanocomplex of claim 1, wherein the nanocomplex selectively targets macrophages overexpressing GLUT (Glucose Transporter) to deliver phytochemicals.

13. A pharmaceutical composition for preventing or treating a muscle disease, comprising the albumin nanocomplex of any one of claims 1 to 12.

14. The pharmaceutical composition for preventing or treating a muscle disease according to claim 13, wherein the muscle disease is caused by an anticancer drug or an inflammatory reaction.

15. 15. The pharmaceutical composition for preventing or treating a muscular disease according to claim 14, wherein the anticancer drug is one or more selected from the group consisting of etoposide, adriamycin, vincristine, cyclophoshamide, ifosphamide, cisplatin, vinorelbine, paclitaxel, docetaxel, gemcitabine, and pemetrexed.

16. 14. The pharmaceutical composition for preventing or treating a muscle disease according to claim 13, wherein the muscle disease is selected from the group consisting of sarcopenia and muscle atrophy.

17. 14. The pharmaceutical composition for preventing or treating a muscle disease according to claim 13, wherein the pharmaceutical composition inhibits muscle loss and promotes proliferation of myoblasts and their differentiation into muscle cells.

18. A food composition for preventing or improving muscle diseases, comprising the albumin nanocomplex of any one of claims 1 to 12.

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