Albumin nanocomposites comprising phytochemicals and compositions for improving muscle diseases comprising same

Abstract

The present invention relates to an albumin nanocomposite comprising a phytochemical, and a composition for improving muscle diseases comprising the same, and more specifically, to an albumin nanocomposite comprising a phytochemical, the present invention relates to an albumin nanocomposite containing phytochemicals, and more specifically, to an albumin nanocomposite containing phytochemicals, which exhibits the effects of inhibiting muscle reduction caused by oxidative stress or inflammatory response and promoting the differentiation of myoblasts into muscle cells. The albumin nanocomposite containing the phytochemicals can target and induce immune cells of active oxygen and inflammatory reaction and effectively deliver the phytochemicals, so that muscle cell atrophy is inhibited and differentiation is promoted by adjusting a muscle reduction signal mechanism induced by the active oxygen and the inflammatory reaction, and further muscle reduction can be improved, so that the curative effect of the albumin nanocomposite is improved. The muscular dystrophy is prevented and treated. Therefore, the composition containing the albumin nanocomposite containing phytochemicals of the present invention can be provided as a composition for improving muscle diseases.

Description

TECHNICAL FIELD

[0001] The present application relates to an albumin nanoparticle complex comprising a phytochemical and a composition for improving muscle disease comprising the same, and more particularly, to an albumin nanoparticle complex comprising a phytochemical for preventing or improving muscle disease by inhibiting oxidative stress or inflammatory response, promoting differentiation of myoblasts into myocytes, and a composition for improving muscle disease comprising the same. BACKGROUND

[0002] In recent years, with the rapid growth of the global aging population, according to the United Nations (UN), the number of people aged 60 and over is expected to exceed 2 billion by 2050. As described above, with the rapid growth of the elderly population aged 65 and over, the quality of life of the elderly in an aging society is increasingly becoming an important issue, and the demand for a longer healthy life is increasing.

[0003] Skeletal muscle is the largest organ in the human body, accounting for 40 to 50% of the total weight of the human body, and is responsible for various functions of the human body and participates in body activities including maintenance of body weight, walking, and energy homeostasis, heat production, sugar and amino acid metabolism. Sarcopenia is one of the typical symptoms of aging, which is accompanied by a decrease in skeletal muscle content and a decrease in muscle function. With the World Health Organization (WHO) giving a disease code to sarcopenia in 2017, experts predict that food companies or pharmaceutical companies will develop a large number of products for preventing and improving sarcopenia.

[0004] Such muscle loss is caused by a combination of factors such as an imbalance between muscle protein synthesis and breakdown, an increase in muscle cell death, and a decrease in muscle regeneration capacity. Muscle reduction and muscle weakness are one of the main physical change symptoms that occur due to aging, and it is known that these changes gradually proceed from the age of 40. Such muscle reduction is accompanied by a decrease in overall physical function, a decrease in the quality of life of individuals, and a high mortality rate in the elderly who lack muscle, and it has been reported that even if they have the same disease, the mortality rate of the elderly with sarcopenia is higher. It has been reported that the causes of sarcopenia include hormone deficiency such as growth hormone and sex hormone, imbalance between muscle protein synthesis and breakdown, inactivity, obesity, increase in inflammatory cytokines, decrease in mitochondrial function, and insulin resistance, etc. In particular, it has been reported that as age increases, obesity and visceral fat increase, and inflammatory cytokines also increase, thereby causing sarcopenia.

[0005] Currently, hormone therapy is a representative therapy for treating sarcopenia, but the effect is poor, and side effects are continuously reported. Many pharmaceutical companies have developed blockers that inhibit or hinder myostatin, and have conducted a number of clinical trials, but all have ended in failure, and thus there is an urgent need for new alternatives.

[0006] It is reported that since causative factors of muscle depletion are various, it is difficult to control muscle depletion by controlling only a single pathway. Thus, in order to improve a clinical success rate, a technology capable of comprehensively controlling muscle weakness pathways is required to be developed, thereby alleviating and treating muscle depletion.

[0007] In order to develop such a new material, Korean Patent No. 10-2216599 discloses a technology of extracting effective components of Semisulcospira libertina and green tea as raw materials for a health functional food for improving muscle depletion, and Korean Patent No. 10-2606636 discloses a technology of providing a Bacillus velezensis strain or a culture solution thereof having an effect of preventing or improving muscle depletion by confirming the effect, and providing the same as a composition for improving muscle depletion.

[0008] However, none of the documents discloses a composition capable of improving muscle disease improvement effects by selectively delivering phytochemicals to tissues inducing muscle depletion. SUMMARY

[0009] TECHNICAL PROBLEM

[0010] An object of the present application is to provide an albumin nano-complex containing phytochemicals.

[0011] Still another object of the present application is to provide a pharmaceutical composition for preventing or treating muscle diseases containing the albumin nano-complex.

[0012] Another object of the present application is to provide a food composition for preventing or improving muscle diseases containing the albumin nano-complex.

[0013] MEANS FOR SOLVING PROBLEM

[0014] The present application provides an albumin nano-complex which is a nano-complex obtained by a click chemistry reaction of albumin to which an azide (N3) or a cyclooctyne functional group is bound, and a delivery substance to which an azide (N3) or a cyclooctyne functional group is bound, and loading phytochemicals on the surface of the albumin, the delivery substance containing a glucosyl group, the delivery substance being bound to the cyclooctyne functional group when the albumin is bound to the azide functional group, and the delivery substance being bound to the azide functional group when the albumin is bound to the cyclooctyne functional group.

[0015] In the present invention, the phytochemical can be one or more selected from the group consisting of phenolic compounds and triterpenes.

[0016] In the present invention, the phenolic compounds can be one or more selected from the group consisting of apigenin, citrusinol, flavone, flavonol, flavanone, flavanol, isoflavone, anthocyanins, stilbenoid, Caffeic acid, p-coumaric acid, Chrysin, tectochrysin, Primetin, acacetin, luteolin, Tangeritin, quercetin, kaempferol, genistein, daidzein, naringenin, hesperetin, cyanidin, catechin, curcumin, epigallocatechingallate, and resveratrol.

[0017] In the present invention, the triterpenes can be 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, beta-sitosterol, campesterol, stigmasterol, lanosterol, and germanicol.

[0018] In the present application, the number of azide or cyclooctyne functional groups introduced into the albumin can be 1 to 14.

[0019] In the present application, the nanocomplex can comprise 4 to 8 glucose groups.

[0020] In the present application, 1 to 20 of the phytochemicals can be loaded.

[0021] In the present application, the azide or cyclooctyne functional group can be bound to the 6th carbon atom of the glucose group.

[0022] In the present application, the delivery substance can further comprise a radioisotope, which can be one or more selected from the group consisting of 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.

[0023] In the present application, the radioisotope can be labeled with a chelator, which can be selected from the group consisting of 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-glutaricacid-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), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,811-tetraacetic acid)), p-SCN-Bn-DTPA (2-(4-isothiocyanatobenzyl)-diethylenetriaminepentaacetic acid At least one of the group consisting of: 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).

[0024] In the present invention, the delivery substance may further comprise a fluorescent substance, and the fluorescent substance may be selected from the group consisting of FNR (Ferrodoxin NADP(+) reductase), cyanine fluorescent substances, TAMRA (tetramethylrhodamine-5-maleimide), Flamm ® One or more selected from the group consisting of a fluorescent substance and ICG (indocyanine green).

[0025] In the present invention, the nanocomplex can deliver phytochemicals by selectively targeting macrophages overexpressing glucose transporter (GLUT).

[0026] The present invention also provides a pharmaceutical composition for preventing or treating muscle diseases, comprising the albumin nanocomplex.

[0027] In the present invention, the muscle disease may be caused by an anticancer agent or an inflammatory response.

[0028] In the present application, the anticancer agent can be one or more selected from the group consisting of Etoposide, Adriamycin, Vincristine, Cyclophoshamide, Ifosphamide, Cisplatin, Vinorelbine, Paclitaxel, Docetaxel, Gemcitabine, and Pemetrexed.

[0029] In the present application, the muscle disease can be selected from the group consisting of muscle reduction and muscle atrophy.

[0030] In the present application, the pharmaceutical composition can inhibit muscle reduction, myoblast proliferation, and promote differentiation into muscle cells.

[0031] The present application also provides a food composition for preventing or improving muscle disease, comprising the albumin nanocomplex.

[0032] Effects of the Invention

[0033] The albumin nanocomplex comprising phytochemicals of the present application can target immune cells inducing reactive oxygen species and inflammatory responses and effectively deliver phytochemicals, thereby inhibiting atrophy and promoting differentiation of muscle cells by regulating oxidative stress, inflammatory responses, or muscle reduction signaling mechanisms induced by anticancer agents, and thus can improve muscle reduction, prevent, and treat muscle dystrophy, and thus the albumin nanocomplex comprising phytochemicals of the present application can be provided as an excellent composition for improving muscle disease. BRIEF DESCRIPTION OF DRAWINGS

[0034] Figure 1 A graph of the albumin nanocomplex loaded with apigenin according to an embodiment of the present application.

[0035] Figure 2 A UV analysis result for confirming the amount of apigenin bound to the surface of glycated albumin according to an embodiment of the present application.

[0036] Figure 3 A UV analysis result for confirming the stability of glycated albumin loaded with apigenin by evaluating the degree of release of the drug over time according to an embodiment of the present application.

[0037] Figure 4 A result of confirming hydrogen peroxide scavenging ability for confirming ROS (reactive oxygen species) scavenging effect of apigenin-glycated albumin according to an embodiment of the present application.

[0038] Figure 5 Results of confirming the oxygen radical scavenging ability in order to confirm the ROS scavenging effect of apigenin-glucosylated albumin according to an embodiment of the present application.

[0039] Figure 6 Results of confirming the intracellular fluorescence expression level of glucose-glycated albumin (Alb-DBCO-Glc) and non-glycated albumin (Alb-DBCO) introduced with FNR648 fluorescence in order to confirm whether M1 macrophages target the glucose-glycated albumin according to an embodiment of the present application.

[0040] FIG. 7 is results of confirming the cell morphology (a) and intracellular / extracellular ROS change (b) after introducing FNR648 fluorescence into glucose-glycated albumin (Glc-Alb), non-glycated albumin (DBCO-Alb), apigenin alone (Apigenin), non-glycated albumin combined with apigenin (AA), and glucose-glycated albumin combined with apigenin (GA) and treating in order to confirm the selective uptake of glucose-glycated albumin according to an embodiment of the present application after inducing the Raw cell line to polarize from M0 to M1 state.

[0041] FIG. 8 is immunofluorescence staining results for confirming the cytotoxicity of apigenin-glucose-glycated albumin according to an embodiment of the present application, Figure 8a to show the fluorescence expression of the nucleus after DAPI (4', 6-diamidino-2-phenylindole) staining, Figure 8b to show a graph of the number of nuclei after DAPI staining.

[0042] FIG. 9 is immunofluorescence staining results for confirming the muscle differentiation promoting effect of apigenin-glucose-glycated albumin according to an embodiment of the present application, Figure 9a to visualize MHC-positive myotubes in green by immunofluorescence and show the nucleus labeled with DAPI in blue, Figure 9b to show a graph of the level of differentiation of mouse myoblasts into myotubes.

[0043] FIG. 10 is immunofluorescence staining results for confirming the improvement effect of apigenin-glucose-glycated albumin according to an embodiment of the present application on muscle atrophy, Figure 10a to show fluorescence expression analysis results of the level of differentiation into myotubes after treating with apigenin alone or apigenin-glucose-glycated albumin in cells in which muscle atrophy was induced with cancer cell culture solution, Figure 10b to show a graph of the level of differentiation into myotubes.

[0044] Figure 11Western blot analysis results were used to confirm the effect of apigenin-glycated albumin on the expression of genes related to muscle production and degradation according to one embodiment of the present invention.

[0045] FIG12 is a result showing the improvement effect of apigenin-glycated albumin on muscle atrophy possibly caused by administration of anticancer drugs according to an embodiment of the present invention. Figure 12a The results of fluorescence expression analysis of the differentiation of mouse myocytes into myotubes after treatment with cisplatin alone or with apigenin-glycated albumin are shown. Figure 12b Figure 2 is a graph showing the level of myotube cell differentiation.

[0046] Figure 13 This is the result of Western blot analysis using proteins extracted from cells in each experimental group in FIG. 12 to confirm the effect of apigenin-glycated albumin on the expression of myogenin that induces muscle growth, according to one embodiment of the present invention.

[0047] Figure 14 The results confirm the feasibility of the injectable formulation of the albumin nanocomplex with GLUT (glucose transporter) targeting capability according to one embodiment of the present invention. These results are fluorescence analysis results confirming the time-dependent changes in the residual amount of the albumin nanoplatform at the injection site by directly injecting the albumin nanocomplex into an animal model.

[0048] Figure 15 The SPECT (single photon emission computed tomography) and PET (positron emission tomography) images of mice injected intramuscularly with apigenin (125I-Api), apigenin and albumin complex (125I-Api / 64Cu-Alb), and apigenin and glycated albumin complex (125I-Api / 64Cu-Glc-Alb) are shown in sequence.

[0049] Figure 16a To confirm the results of fluorescence expression analysis of the differentiation level of myotubes in cells induced by muscle atrophy using cancer cell culture medium after treatment with ursolic acid alone or ursolic acid-glycated albumin, Figure 16b Figure 2 is a graph showing the level of myotube cell differentiation.

[0050] Figure 17a To confirm the results of fluorescence expression analysis of the differentiation level of myotubes in cells induced with muscle atrophy using cancer cell culture medium after treatment with soyasapogenol B alone or with soyasapogenol B-glycosylated albumin, Figure 17b Figure 2 is a graph showing the level of myotube cell differentiation.

[0051] Figure 18aTo confirm the results of fluorescence expression analysis of the level of differentiation into myotube cells after treatment with citral alone or citral-sialbumin after inducing muscle atrophy in cells with cancer cell culture solution, Figure 18b A graph showing the level of myotube cell differentiation. DETAILED DESCRIPTION

[0052] Hereinafter, a specific embodiment of the present application will be described in more detail. Unless otherwise defined, all technical and scientific terms used in the present specification have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Generally, the terms used in the present specification have the same meanings as commonly understood by one of ordinary skill in the art and are not to be interpreted in an idealized or overly formal sense.

[0053] The present application relates to an albumin nano-complex comprising a phytochemical and a composition for improving muscle disease comprising the same.

[0054] The phytochemical is a compound word of "phyto" indicating plant and "chemical" indicating chemistry, and refers to a plant-derived chemical substance having a physiological activity that contributes to health.

[0055] The present application can specifically regulate oxidative stress and inflammatory responses induced by anticancer agents or immune cells by loading a phytochemical on an albumin nano-platform, and exhibit an effect of improving muscle disease associated with oxidative stress and inflammatory responses.

[0056] In the present application, the phytochemical can be a phenolic compound or a triterpene.

[0057] The phenolic compound that can be used in the present application is preferably a flavonoid, a stilbenoid, Caffeic acid, p-coumaric acid, curcumin, resveratrol, etc.

[0058] The flavonoid is a class of secondary metabolites of plants or fungi, and generally has a 15-carbon skeletal structure consisting of two benzene rings and one heterocyclic ring. More than 5000 natural flavonoids are known, and researches using various activities thereof are continuously ongoing.

[0059] As the flavonoid compound, apigenin, citrusinol, flavone, flavonol, flavanone, flavanol, isoflavone, anthocyanins, Chrysin, tectochrysin, Primetin, acacetin, luteolin, Tangeritin, quercetin, kaempferol, genistein, daidzein, naringenin, hesperetin, cyanidin, catechin, epigallocatechin gallate, etc. can be used, and apigenin and citrusinol are preferred.

[0060] Apigenin is a flavonoid compound, which is a yellow pigment contained in various fruits and vegetables, and is known to be abundant in parsley, chamomile, celery, and carrots. For several centuries, this apigenin has been used in the form of chamomile tea or directly applied to the skin, but recently it has been found to be a molecule capable of regulating various intracellular mechanisms, and in particular, it has been reported to inhibit inflammation and oxidative stress and promote carbohydrate metabolism.

[0061] As shown in Figure 1 The present application can provide an albumin nanocomplex and a composition comprising the same by constructing a form in which apigenin is loaded on an albumin-based nanocomplex, by targeting activated M1 type inflammatory macrophages around an acute inflammation or cancer site and precisely delivering apigenin to the target site, and thus can inhibit muscle reduction induced by oxidative stress or inflammation, promote muscle cell differentiation, and thus improve muscle disease.

[0062] Citrusinol is a flavonoid compound found in plants such as Citrus unshiu and Phyllodinium pulchellum. According to the present application, an albumin nanocomplex can be provided, which can improve muscle disease by loading citrusinol on an albumin nanocomplex platform.

[0063] Triterpenes are a general term for compounds having a carbon skeleton composed of 30 carbon atoms among terpenes, which are the main components of plant oils, most of which have 4 or 5 cyclic structures. Triterpenes are known to have anti-inflammatory, antioxidant, and anticancer activities.

[0064] In the present application, as the triterpenes, ursolic acid, soyasapogenol A, soyasapogenol B, ursane, oleanane, lupane, saponin, sterol, oleanolic acid, lupeol, corosolic acid, maslinic acid, betulinic acid, ginsenoside, beta-sitosterol, campesterol, stigmasterol, lanosterol, germanicol, and the like can be used, and ursolic acid and soyasapogenol B are preferred.

[0065] The phytocompounds delivered by the albumin nanocomplex of the present application can exhibit an effect of improving muscle diseases associated with oxidative stress and inflammatory responses by specifically modulating the oxidative stress and inflammatory responses induced by anticancer agents or immune cells. In particular, the albumin nanocomplex of the present application not only improves the low water solubility and delivery ability of natural active ingredients such as apigenin, but also effectively scavenges ROS (Reactive oxygen species) in cells, thereby exhibiting a more superior effect than when the phytocompounds are used alone, and the effect has been experimentally confirmed at the immune cell level. Also, it was confirmed that the expression amount of muscle cell differentiation-related proteins was increased by treating muscle cells, and muscle cell differentiation was promoted, thereby confirming an effect of relieving inflammation and improving muscle function.

[0066] An embodiment of the present application provides an albumin nano-complex, which is a nano-complex of albumin to which an azide (N3) or a cyclooctyne functional group is bound by a click chemistry reaction, and a delivery substance to which an azide (N3) or a cyclooctyne functional group is bound, a phytochemical being loaded on the surface of the albumin, the delivery substance including a glucosyl group, the delivery substance being bound to the cyclooctyne functional group when the albumin is bound to the azide functional group, the delivery substance being bound to the azide functional group when the albumin is bound to the cyclooctyne functional group.

[0067] In the present application, albumin is one of proteins constituting basic substances of cells, and is a protein that exists in a large amount in blood and is produced by a liver. Albumin is the smallest protein in simple proteins existing in nature in terms of molecular weight. Serum albumin in blood has a function of maintaining and restoring plasma volume, and thus is used for preventing shock caused by excessive bleeding as well as for surgical and burn treatment. Also, it is known to have an oxygen delivery capacity similar to that of hemoglobin.

[0068] The albumin can include all formulatable albumins, preferably derived from human plasma, or can be a recombinant human serum albumin prepared by genetic engineering, but is not limited thereto. The genetic information of the albumin according to the present application can be obtained from a known database such as NCBI GenBank (National Center for Biotechnology Information, a global public nucleotide sequence database).

[0069] In the present application, the number of amino groups (-NH2) exposed on the surface of the albumin can be 15 to 30.

[0070] In one embodiment of the present application, in order to prepare a nano-complex based on albumin having biocompatibility, which can target living tissues and deliver a delivery substance such as a drug or a fluorescent substance, and exhibit a therapeutic effect, a representative human serum albumin (HSA) is used, and a cyclooctane (Cyclooctyne) or an azide (N3) as a functional group on one side of click chemistry is introduced to the surface of the human serum albumin under a reaction condition in which the denaturation of the HSA is minimized.

[0071] In the present application, azide (N3) is a reactive group consisting of three nitrogen atoms, known to have high reactivity, particularly in 1,3-dipolar cycloaddition reaction as a Cu-Free click chemistry, as an electron donor to generate triaza-5-membered ring.

[0072] Cyclooctyne functional group is an eight-membered aliphatic ring containing a triple bond subject to ring strain, and particularly, known to generate triaza-5-membered ring as an electron acceptor in 1,3-dipolar cycloaddition reaction as a Cu-Free click chemistry. Due to the structural characteristics of cyclooctyne, i.e., triple bond structure subject to ring strain, click chemistry can be achieved even without Cu(I) catalyst.

[0073] The cyclooctyne functional group can be one or more selected from the group consisting of 4-cyclooctyn-1-yl (4-COO) ), 3-cyclooctyn-1-yl (3-COO) ), 2-cyclooctyn-1-yl (2-COO) ), monofluorinated cyclooctyne (MOFO) ), difluorinated cyclooctyne (DIFO) ), dimethoxyazacyclooctyne (DIMAC) ), dibenzocyclooctyne (DIBO) ), azadibenzocyclooctyne (ADIBO) ), and biarylazacyclooctynone (BARAC) ), but not necessarily limited thereto.

[0074] In the present application, the number of click reaction functional groups (azide or cyclooctyne functional group) introduced into the albumin surface is preferably 1 to 14, more preferably 9 to 12. In this case, various delivery substances can be injected into the human body in excess, thereby increasing the possibility of being absorbed by the target site. On the contrary, if the number of click reaction functional groups exceeds the range, it can be immediately taken up by the liver after being injected into the body, resulting in a possibility of limiting its uptake by other target disease sites. The number of click reaction functional groups introduced into the albumin surface can be adjusted depending on the reaction ratio of albumin to azide-NHS (N-hydroxysuccinimide ester) or cyclooctyne-NHS.

[0075] The solution combined with azide (N3) or the albumin solution combined with cyclooctyne functional group can be obtained by the following steps: step (a), dissolving albumin in a phosphate-buffered saline (PBS) solution; step (b), dissolving azide-NHS or cyclooctyne-NHS in DMSO (dimethyl sulfoxide); and step (c), mixing the resulting solutions and reacting at 20 to 37°C for 30 minutes to 1 hour.

[0076] In step (a), the pH of the phosphate-buffered saline (PBS) solution can be 6.8 to 7.6, preferably 7.0 to 7.4.

[0077] In step (b), the amount of DMSO used to prepare the azide-NHS solution or cyclooctyne-NHS solution can be 2% (v / v) or less of the total reaction solution.

[0078] In step (c), the mixing molar ratio of albumin to azide-NHS or cyclooctyne-NHS can be 1:1 to 1:25.

[0079] When the albumin solution and the azide-NHS solution are mixed in step (c), the functional group combined with the albumin can be an azide functional group, and when the albumin solution and the cyclooctyne-NHS solution are mixed in step (c), the functional group combined with the albumin can be a cyclooctyne functional group.

[0080] In an embodiment of the present application, a human serum albumin (HSA) solution and an ADIBO-NHS (azadibenzocyclooctyne-N-hydroxysuccinimide ester) solution are mixed and reacted at 37°C for 30 minutes to prepare HSA-ADIBO (human serum albumin-azadibenzocyclooctyne).

[0081] The number of click reaction functional groups (azide or cyclooctyne functional groups) introduced into the surface of the albumin is preferably 1 to 14, and more preferably 10 to 12. In this case, various delivery substances can be injected into the human body in excess, thereby increasing the possibility of being absorbed by the target site. In contrast, if the number of click reaction functional groups exceeds the range, it can be immediately taken up by the liver after being injected into the body, resulting in a possibility of limiting its uptake by other target disease sites. The number of click reaction functional groups introduced into the surface of the albumin can be adjusted depending on the reaction ratio of the albumin to azide-NHS or cyclooctyne-NHS.

[0082] The albumin nanocomplex of the present application is obtained by mixing the above-described solution containing albumin to which an azide (N3) or cyclooctyne functional group is bound, and a solution containing a delivery substance to which an azide (N3) or cyclooctyne functional group is bound, and performing a click chemistry reaction, wherein the click chemistry reaction can be a Cu-free catalyst click chemistry reaction.

[0083] In one embodiment of the present application, since the azide functional group used as a click chemistry functional group is an electron donor, and the cyclooctyne functional group is an electron acceptor, when the functional group bound to the albumin is an azide functional group, the functional group bound to the delivery substance is preferably a cyclooctyne functional group, and when the functional group bound to the albumin is a cyclooctyne functional group, the functional group bound to the delivery substance is preferably an azide functional group.

[0084] In the present application, the albumin nanocomplex can contain a phytochemical on the surface of the albumin.

[0085] In this case, the phytochemical loaded on the nanocomplex is a concept including the above-described listed phytochemicals and their derivatives. For example, the derivative of apigenin can mean that the part of oxygen or hydrogen contained in apigenin is substituted with another element or a substituent.

[0086] The phytochemical is loaded on the surface of the albumin, and it is analyzed that there is a pocket that can specifically load the phytochemical on the surface of the albumin, so that the phytochemical can be stably loaded and form a nanocomplex.

[0087] The nanocomplex of the present application can improve muscle diseases by precisely delivering the phytochemical loaded on the surface of the albumin to the target site, thereby inhibiting muscle reduction induced by oxidative stress, inflammatory response, or administration of an anticancer agent, promoting muscle cell differentiation.

[0088] In particular, the apigenin-loaded albumin nanoparticle can play a synergistic role in improving muscle disease activity when used in the form of a glycated albumin.

[0089] The nanoparticle can contain 1 to 20 phytochemicals on the surface of the albumin, preferably 2 to 10, and more preferably 4 to 8 phytochemicals.

[0090] Since the albumin nanoparticle of the present application has a plurality of click reaction functional groups, various delivery substances can be bound to the plurality of reactive groups.

[0091] In the present application, the delivery substance refers to a substance that binds to albumin and is delivered into a living organism, and in the present application, it contains a glucose group, the structure of which is shown in the following Formula 1.

[0092] Formula 1

[0093]

[0094] The albumin nanoparticle of the present application uses glucose as a target ligand, targeting the glucose transporter (GLUT, Glucose Transporter) overexpressed in M1 macrophages, thereby delivering phytochemicals.

[0095] In the present application, the albumin nanoparticle can contain 4 to 8 glucose groups, preferably 5 to 7. In this case, the nanoparticle injected into the body can remain in the injection site or blood for a long time, thereby increasing the possibility of targeting the target site, however, if the number of glucose groups exceeds 8, it will be immediately taken up by the liver after being injected into the body, and thus the targeting of the target disease site can be limited.

[0096] In the present application, the glucose group as a delivery substance can be bound by a click chemistry reaction between an azide or cyclooctyne functional group bound to albumin and an azide or cyclooctyne functional group bound to the glucose group.

[0097] In this case, GLUT overexpressed on the surface of M1 cells recognizes the OH functional group at the 1st, 3rd, and 5th positions of glucose, thereby obtaining glucose as an energy source. In the present application, in order to more effectively target M1 macrophages, the 1st, 2nd, and 6th carbon positions of glucose can be used to bind to albumin, and among them, the 6th carbon position is most preferred.

[0098] For example, the glucose group bound to the 1st, 2nd, or 6th carbon position of azide has a chemical structure shown in the following Formulas 2, 3, and 4, respectively.

[0099] Formula 2

[0100]

[0101] Formula 3

[0102]

[0103] Formula 4

[0104]

[0105] The nanocomplex in which the albumin is bound to the 6th carbon of glucose can exhibit 3 times and 2 times higher GLUT targeting ability, respectively, compared to the nanocomplex in which the albumin is bound to the 1st and 2nd carbons of glucose.

[0106] In the present application, the nanocomplex can further comprise one or more substances as delivery substances, in which case, the plurality of substances can be simultaneously subjected to the click chemistry reaction with the albumin to which the azide or cyclooctyne functional group is bound, or the plurality of substances can be sequentially subjected to the click chemistry reaction, respectively.

[0107] In the present application, as the delivery substance, a radioisotope, which refers to an element having the same atomic number but different atomic mass, among isotopes, is used. Such a radioisotope can be used as an important marker for diagnosing a disease or confirming pharmacodynamics, using the characteristic of decaying radioactivity by emitting gamma rays or other subatomic particles. The radioisotope used as the marker in the present application can be used without limitation from among the radioisotopes well known in the art, and can be selected from the group consisting of 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, preferably 11 C, 18 F, 64 Cu, 67 Ga, 68 Ga, 89 Zr, 99m Tc, 111 IN and 123 I, but not necessarily limited thereto.

[0108] In the present invention, the radioisotope is labeled with a chelator that functions to link the radioisotope to albumin, for example, can be selected from the group consisting of 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-glutaricacid-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), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), TACN (1,2,4-triazacyclohexane), TETA (1,4,8,11-tetraazacycl11-tetraacetic acid)), p-SCN-Bn-DTPA (2-(4-isothiocyanatobenzyl)-diethylenetriaminepentaacetic acid 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), but the present invention is not necessarily limited thereto.

[0109] In the present invention, the delivery substance may further include a fluorescent substance.

[0110] Fluorescent substances refer to substances that emit visible light at a specific wavelength. In the present invention, fluorescent substances include all fluorescent substances that can be combined with the albumin nanocomplex to confirm the location of the nanocomplex.

[0111] Specifically, as fluorescent substances that can be used as markers in the present invention, radioactive isotopes known in the art can be used without limitation, and the following fluorescent substances can be used: rhodamines, including rhodamine, tetramethylrhodamine (TAMRA), etc.; fluoresceins, including fluorescein, fluorescein isothiocyanate (FITC) and fluorescein amide (FAM), etc.; boron-dipyrromethene (bodipy); AF (alexa fluor); and cyanines, including Cy3, Cy5, Cy7, indocyanine green, etc., but not limited thereto.

[0112] Another embodiment of the present invention provides a pharmaceutical composition for preventing or treating muscle diseases, comprising an albumin nanocomplex containing phytochemicals.

[0113] In the present invention, the muscle disease may be a muscle disease caused by an anticancer agent or an inflammatory response.

[0114] Specifically, the albumin nanoparticle complex comprising a phytochemical of the present application can inhibit muscle loss by modulating the signaling mechanism of anticancer agent administration and immune cell-induced oxidative stress or inflammatory response-induced muscle weakness, and induce muscle mass increase by promoting myocyte differentiation.

[0115] According to an embodiment of the present application, the apigenin-loaded glycated albumin nanoparticle complex selectively targets inflammatory M1-type inflammatory microphages in mouse myocytes, thereby inhibiting myocyte loss induced by inflammation and promoting myoblast differentiation, while at the same time, inhibiting myocyte loss and atrophy induced by anticancer agent administration and promoting myocyte differentiation, thereby exhibiting an effect of improving muscle loss and muscular dystrophy.

[0116] In particular, the cell group administered with the apigenin-loaded glycated albumin nanoparticle complex exhibited a significant decrease in ROS compared to the cell group administered with apigenin alone, and exhibited an effect of inhibiting differentiation of macrophages into inflammatory type. Also, in myocytes in which muscle atrophy was induced by cancer cell culture solution and anticancer agent, the improvement effect of the cell group administered with the apigenin-loaded glycated albumin nanoparticle complex was confirmed to be significantly increased compared to the apigenin alone administration group, and thus it can be confirmed that the apigenin and the glycated albumin nanoparticle complex of the present application exert a synergistic effect.

[0117] Also, since it was confirmed that the albumin nanoparticle complex comprising apigenin can inhibit expression of myocyte decomposition genes induced by immune cells and anticancer agent administration, and increase expression of genes inducing differentiation of myoblasts into myocytes, the pharmaceutical composition of the present application containing the apigenin-containing albumin nanoparticle complex can prevent or treat muscle diseases by inhibiting muscle loss, myoblast proliferation, and promoting differentiation into myocytes.

[0118] In the present application, the anticancer agent can be one or more selected from the group consisting of Etoposide, Adriamycin, Vincristine, Cyclophoshamide, Ifosphamide, Cisplatin, Vinorelbine, Paclitaxel, Docetaxel, Gemcitabine, and Pemetrexed.

[0119] Also, in another embodiment of the present application, it is confirmed that ursolic acid, soyasapogenol B, and citral are composed of the same albumin nano-complex as apigenin, and thus can inhibit muscle reduction, myoblast proliferation, and promote myocyte differentiation, and in particular, it is confirmed that these substances can exert more excellent synergistic effects than when used alone.

[0120] In the present application, the muscle disease can be selected from the group consisting of muscle reduction and muscular atrophy.

[0121] In the present application, the muscle reduction or muscular atrophy can be interpreted to include not only the dictionary meaning of muscle reduction (sarcopenia) or muscular atrophy (muscular atrophy), but also a muscle disease caused by or induced by muscle reduction or muscular atrophy, such as sarcopenic obesity, myopathy, muscular dystrophy, muscular injury, myasthenia, myoneural conductive disease, nerve injury, amyotrophic lateral sclerosis (ALS), atony, myotonia, cachexia, etc.

[0122] For example, the muscular dystrophy can be meant by diabetic amyotrophy, spinal muscular atrophy, etc., the muscular dystrophy can be meant by Duchenne muscular dystrophy, Becker muscular dystrophy, limb girdle muscular dystrophy, facioscapulohumeral muscular dystrophy, oculopharyngeal muscular dystrophy, Myotonic dystrophy, etc., and the myopathy can be meant by inflammatory myopathy such as polymyositis and dermatomyositis, endocrine myopathy, toxic myopathy, metabolic myopathy, mitochondrial myopathy, congenital myopathy, etc.

[0123] The pharmaceutical composition can further include a pharmaceutically acceptable carrier, excipient, or diluent appropriate for a conventional method. The pharmaceutically acceptable carrier is a carrier commonly used in the preparation of a pharmaceutical composition, including lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginic acid, gelatin, calcium silicate, microcrystalline cellulose, polyvinyl pyrrolidone, cellulose, water, syrup, methyl cellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil, but is not limited thereto.

[0124] The pharmaceutical composition of the present application can further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc., in addition to the above ingredients. As to the suitable pharmaceutically acceptable carrier and dosage form, the method disclosed in Remington's Pharmaceutical Sciences (19th ed., 1995) can be preferably used according to each ingredient.

[0125] The pharmaceutical composition of the present application can be administered orally or parenterally, and the parenteral administration includes intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc.

[0126] Dosage forms for oral administration include, for example, tablets, pills, hard capsules, soft capsules, liquid preparations, suspensions, emulsions, syrups, granules, etc., and these dosage forms can contain, in addition to the effective ingredient, diluents (e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and / or glycine), lubricants (e.g., silica, talc, stearic acid, its magnesium or calcium salt, and / or polyethylene glycol). Also, the tablets can contain binders, e.g., magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidone, if necessary, disintegrating agents or effervescent mixtures, e.g., starch, agar, alginic acid or its sodium salt, and / or an absorbent, a coloring agent, a flavoring agent, and a sweetening agent. The dosage forms can be prepared by conventional mixing, granulation or coating methods.

[0127] Also, representative examples of dosage forms for parenteral administration are injection preparations, and as solvents for injection preparations, water, Ringer's solution, isotonic saline or suspensions, etc. can be exemplified.

[0128] The injection preparation can use a sterile, non-volatile oil as a solvent or suspending medium, and any non-irritating, non-volatile oil, including mono- and di-glycerides, can be used for this purpose. Also, the injection preparation can use a fatty acid, such as oleic acid.

[0129] The composition of the present application is administered in a pharmaceutically effective amount. In the present application, "pharmaceutically effective amount" means a dose sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level can be determined depending on the type and severity of the patient's disease, the activity of the drug, the sensitivity to the drug, the administration time, the administration route and the excretion rate, the treatment period, the drugs used simultaneously, and other factors known in the medical field. The composition of the present application can be administered as a single therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered once or multiple times. Considering all the above factors, it is important to administer the minimum dose to achieve the maximum therapeutic effect without side effects, and this can be easily determined by a person with ordinary knowledge.

[0130] Specifically, the effective dose of the composition of the present application can vary depending on the age, sex, and body weight of the patient, and is generally administered at a dose of 0.001 to 150 mg, preferably 0.01 to 100 mg, per 1 kg of body weight, once or every other day, or 1 to 3 times a day. However, since the dose can be increased or decreased depending on the administration route, sex, body weight, age, etc., the dose does not limit the scope of the present application in any way.

[0131] Also, the present application provides a food composition for preventing or improving muscle disease containing a nanocomplex containing a phytochemical. The food of the present application can be in the meaning of food and feed additives including for humans as well as animals.

[0132] In still another embodiment of the present application, the food composition can further comprise one or more additives selected from the group consisting of organic acids, phosphates, antioxidants, lactoseryl casein, dextrin, glucose, sugar, and sorbitol. The organic acid can be, but is not limited to, citric acid, fumaric acid, adipic acid, lactic acid, or malic acid, the phosphate can be, but is not limited to, sodium phosphate, potassium phosphate, acid pyrophosphate, or polyphosphate (polyphosphates), the antioxidant can be, but is not limited to, a natural antioxidant such as polyphenol, catechin, alpha-tocopherol, rosemary extract, licorice extract, chitosan, tannin, or phytic acid.

[0133] In another embodiment of the present application, the food composition can further comprise various nutrients, vitamins, minerals (electrolytes), flavoring agents (e.g., synthetic flavoring agents and natural flavoring agents), coloring agents, and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerol, alcohol, carbonating agents used in carbonated beverages, etc. in addition to the effective ingredient. Also, the food composition of an embodiment of the present application can contain pulp for preparing natural fruit juice, fruit juice beverages, and vegetable beverages.

[0134] According to an embodiment of the present application, the dosage form of the food composition can be, but is not limited to, solid, powder, granule, tablet, capsule, liquid, or beverage form.

[0135] Also, the food composition can be used to manufacture foods such as, but not limited to, cookies, sugars, ice cream products, dairy products, meat products, fish products, tofu, edible oils, noodles, teas, beverages, special nutritional foods, health foods, seasoning foods, ice, ginseng products, kimchi pickled foods, dried foods, fruits, vegetables, dried fruits or vegetables, sliced products, fruit juices, vegetable juices, mixed juices thereof, potato chips, noodles, livestock processed foods, aquatic processed foods, dairy processed foods, fermented dairy products, legume foods, cereal foods, microorganism fermented foods, candies and baked foods, seasonings, processed meats, acidic beverages, licorice, herbs, etc.

[0136] As described above, the albumin nanoparticle complex of the present application containing phytochemicals effectively delivers phytochemicals by targeting immune cells inducing reactive oxygen species and inflammatory responses, thereby regulating oxidative stress and inflammatory response-induced muscle loss signaling mechanisms, and thus inhibiting atrophy and promoting differentiation of muscle cells, so that effects of improving muscle loss and preventing and treating muscular dystrophy can be obtained.

[0137] Example

[0138] The present application will be described in more detail by the following examples. However, for illustrative purposes of the present application, the examples describe some experimental methods and configurations, but the scope of the present application is not limited to the examples.

[0139] Preparation Example 1: Preparation of Apigenin-glycated albumin complex

[0140] 1-1. Preparation of glycated albumin complex

[0141] The glycosylated albumin to which glucose is bound was prepared by the following method.

[0142] Albumin was dissolved in a phosphate buffer solution (PBS, pH 7.4) at a concentration of 40 mg / mL, and the albumin solution was aliquoted into vials at 20 mg / 0.5 mL. DBCO-NHS was dissolved in DMSO (10 mg / 50 μL) and added to the albumin solution to make 500 μL.

[0143] Albumin and DBCO-NHS were reacted at a ratio of 1:14 at room temperature for 6 hours, and purified with amico N Ultra-0.5 centrifugal filter tubes to obtain DBCO-albumin (DBCO-Alb).

[0144] To synthesize glucose-albumin, 6-azido-glucose was dissolved in distilled water at a concentration of 3 mg / mL, and then added to the DBCO-albumin (DBCO-Alb) solution (4 mg / 0.5 mL). After incubation at room temperature for 1 hour, purification was performed with a centrifugal filter tube.

[0145] Then, the absorbance at 280 and 309 nm was confirmed by ultraviolet-visible (UV-Vi) spectrophotometry, thereby confirming the tendency of glucose-DBCO-Alb (Alb-DBCO-Glc). To measure the absorbance of albumin, each albumin sample was diluted to 2 mg / mL and used, and matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF MS) was performed to determine the amount of ADIBO and glucose bound to albumin (functionalization degree = DOF).

[0146] Subsequently, the number of DBCO molecules on the surface of the albumin and the number of glucose molecules introduced by the click reaction were measured using UV-visible spectrophotometry and matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis. To prepare DBCO 11 -Alb-FL and Glc 6 -DBCO 11 -Alb-FL, N3-FNR648 (10 nmol / 1 μL, dissolved in DMSO) was added to DBCO-Alb and Glc-DBCO-Alb (30 nmol / 500 μL), and incubated at 4°C for 30 minutes, and then purified using a centrifugal filter.

[0147] The number of DBCO functional groups introduced in HSA-DBCO was quantitatively analyzed using UV-visible spectrophotometry, and the result was confirmed to be 12.8 to 13.1, and when quantitatively analyzed using matrix-assisted laser desorption ionization time-of-flight mass spectrometry, it was confirmed to be 11.1 to 11.5, and in this case, the number of glucose binding was confirmed to be 5.4 to 7.8.

[0148] 1-2. Preparation of glycated albumin complex loaded with apigenin

[0149] It was intended to load apigenin onto the glycosylated albumin prepared in Preparation Example 1-1.

[0150] 0.5, 1, 2, 4, 8, 16, and 32 equivalents of apigenin were mixed with albumin on the basis of albumin, and after reacting for 0.5 to 2 hours, purified using a filter, and then evaluated by UV. The result is shown in Table 1, and the apigenin characteristic peak between 350 nm and 380 nm was saturated after 16 equivalents, so it can be confirmed that the level of apigenin binding to each albumin was 8 to 16. Figure 2

[0151] Then, the stability of the complex was confirmed by evaluating the degree of drug release over time. The glycosylated albumin loaded with apigenin was prepared immediately after setting the time to 0 hours, and the albumin was sampled after 10 minutes, 30 minutes, 1 hour, 1 hour and 30 minutes, 3 hours, 6 hours and 50 minutes, 17 hours, 24 hours, and 88 hours, respectively, and then the degree of release was confirmed by UV, and the result is shown in Table 2. Figure 3

[0152] Referring to Table 2, it was confirmed that the amount of drug released was about 20% after 88 hours, and thus it can be confirmed that apigenin has been stably loaded on glycosylated albumin to form an apigenin-glycosylated albumin complex. Figure 3

[0153] Experimental Example 1: Confirmation of reactive oxygen species (ROS) scavenging effect ​​​

[0154] To confirm the ROS scavenging effect of the apigenin-sugarated albumin complex prepared in Preparation Example 1, a hydrogen peroxide and oxygen radical scavenging ability evaluation experiment was performed, and the results are shown in Figure 4 and Figure 5

[0155] In the experiment, apigenin alone (API), albumin combined with apigenin (AA), and apigenin-sugarated albumin (GA) combined with apigenin were compared and evaluated, and in this case, the concentration of apigenin was 12.5 μM.

[0156] Referring to the hydrogen peroxide scavenging effect confirmed in Figure 4 , the hydrogen peroxide scavenging ability showed the same level of effect in all groups, and thus it can be confirmed that the structure of the complex formed with albumin does not affect the oxidative stress scavenging ability of apigenin.

[0157] Also, in the oxygen radical scavenging ability evaluation experiment Figure 5 , a similar level of effective oxygen radical scavenging effect can be confirmed.

[0158] Thus, it can be confirmed that the apigenin-sugarated albumin has ROS scavenging ability, and even if a complex is formed with albumin, the efficacy of apigenin can be maintained.

[0159] Experimental Example 2: Confirmation of M1 macrophage targeting and inhibitory effect

[0160] 2-1. Confirmation of M1 macrophage targeting ability of glycated albumin

[0161] The sugarated albumin of Preparation Example 1-1 was prepared using glucose as a target ligand to selectively target glucose transporters (GLUT, Glucose Transporter) overexpressed in M1 type macrophages.

[0162] To confirm whether the albumin sugarated by glucose can actually selectively target M1 type macrophages, the Raw cell line was polarized into M0 and M1 states, and then treated with FNR648 fluorescence in sugarated albumin (Alb-DBCO-Glc) and non-sugarated albumin (Alb-DBCO).

[0163] Referring to the graph showing the targeting ability of each albumin treated with cells Figure 6 , the uptake rate of sugarated albumin in M1 was more than 4 times higher than that of non-sugarated albumin.

[0164] ​Specifically, it was confirmed that there was no significant difference in the uptake rate of the two types of albumin in M0, and there was no significant difference between the groups, but the fluorescence expression of glycated albumin was significantly increased in M1, and thus it was confirmed that the glycated albumin was a platform that could be specifically taken up by M1.

[0165] 2-2. Confirmation of M1 macrophage inhibition of apigenin-glycated albumin

[0166] As in the previous experiment, it was confirmed whether apigenin-glycated albumin could selectively inhibit M1-type macrophages.

[0167] By the same method as in Experimental Example 2-1, after inducing the Raw cell line to polarize from M0 to M1, FNR648 fluorescence was introduced in glycated albumin (Glc-Alb), non-glycated albumin (DBCO-Alb), apigenin alone, non-glycated albumin combined with apigenin (D / A), and glycated albumin combined with apigenin (G / A) to perform treatment. Subsequently, the fluorescence expression was confirmed, and the changes in cell morphology and intracellular / extracellular ROS were confirmed, and the results are shown in Figure 7a and 7b

[0168] As a result, ROS reduction was confirmed in the apigenin alone administration group and the experimental groups treated with albumin combined with apigenin (D / A and G / A), and it was confirmed that the Raw cell line maintained the MO type in terms of morphology.

[0169] In particular, compared to the other experimental groups, glycated albumin combined with apigenin (G / A) significantly reduced ROS, showing an effect of inhibiting differentiation into M1 type, and thus it was confirmed that apigenin and glycated albumin exert a synergistic effect.

[0170] Experimental Example 3: Confirmation of cytotoxicity of apigenin-glycated albumin

[0171] The cytotoxicity of apigenin-glycated albumin was confirmed by performing immunofluorescence staining.

[0172] After culturing mouse myoblast C2C12, apigenin alone and apigenin-glycated albumin were treated at the same concentration of 2.5 μM and 5 μM, respectively, and after culturing for 96 hours, the nuclei were stained with DAPI, and then the fluorescence expression was confirmed as shown in Figure 8a , and the number of stained nuclei was counted, and the results are shown in Figure 8b

[0173] Referring to Figure 8b ​​It was confirmed that the group treated with apigenin alone or apigenin-glycated albumin did not exhibit cytotoxicity.

[0174] Experimental Example 4: Confirmation of effect of apigenin-glycated albumin on promoting myocyte differentiation

[0175] To confirm the effect of apigenin-glycated albumin in promoting muscle differentiation, immunofluorescence staining was performed.

[0176] After culturing mouse myoblast C2C12, apigenin alone and apigenin-glycated albumin were treated at the same concentration of 2.5 μM and 5 μM, respectively, and myotube cell differentiation was performed for 48 hours.

[0177] Then, the cells were fixed with 4% formaldehyde solution, and myosin heavy chain (MHC, Developmental Studies Hybridoma Bank, Iowa City, Iowa, USA) and 4', 6-diamidino-2-phenylindole dihydrochloride (DAPI, Sigma Aldrich, MO, USA) were used as a primary antibody and reacted overnight at 4°C, and a secondary antibody was attached for 1 hour at room temperature.

[0178] Subsequently, as shown in Figure 9a , the immunofluorescence results of MHC-positive myotubes were shown in green, and the DAPI-labeled nuclei were shown in blue. Also, the total number of nuclei was confirmed to differentiate mouse myoblasts into myotubes using a mathematical formula (Fusion Index = number of nuclei in myotube cells / myocytes), and the results are shown in Figure 9b .

[0179] Referring to Figure 9a and 9b , it was confirmed that the group treated with apigenin-glycated albumin showed increased myotube cell differentiation compared to the group treated with apigenin alone, and in particular, the group treated with 2.5 μM apigenin-glycated albumin showed a 15% increase in myotube cell differentiation compared to the group treated with the same concentration of apigenin alone. It was thus confirmed that apigenin-glycated albumin can promote muscle generation by increasing the differentiation of myoblasts.

[0180] Experimental Example 5: Improvement effect of apigenin-glycated albumin on muscle atrophy

[0181] To confirm the improvement effect of apigenin-glycated albumin on muscle atrophy, immunofluorescence staining was performed using the same method as in Experimental Example 3.

[0182] First, mouse myoblast C2C12 was cultured, and then the medium was replaced with cancer cell medium (CCM) in which mouse colon carcinoma cell CT26 was cultured, and muscle atrophy was induced, and apigenin was treated alone at a concentration of 2.5 μM and 5 μM or apigenin-glycated albumin was treated, and after 48 hours of culture, the fluorescence expression of the cells was confirmed, thereby confirming the differentiation level of myotube cells.

[0183] Referring to the level of differentiation of atrophy-induced myoblast into myotube cells after treatment with apigenin or apigenin-glycated albumin Figure 10a and 10b Compared to the experimental group in which apigenin was treated alone, in the experimental group in which apigenin-glycated albumin was treated, it was confirmed that the differentiation of atrophy-induced myoblast into myotube cells was increased. In particular, compared to the experimental group in which apigenin was treated alone at the same concentration, the experimental group in which apigenin-glycated albumin was treated at 2.5 μM showed a 15% increase in myotube cell differentiation, and thus it was confirmed that apigenin-glycated albumin can exhibit an effect of improving muscle atrophy by increasing the differentiation of myoblast.

[0184] On the other hand, to confirm the effect of apigenin-glycated albumin on the expression of muscle generation and decomposition-related genes, proteins were extracted from the cells of the respective experimental groups, and Western blot analysis was performed, and then the protein expression level of each gene was confirmed, and the results are shown in Table 2. Figure 11

[0185] Referring to Figure 11 In the cells atrophied by cancer cell medium (CCM), regardless of the experimental group in which apigenin was treated alone or the experimental group in which apigenin-glycated albumin was treated, MHC expression was increased, and it was confirmed that the muscle cells atrophied by CCM were improved. Also, it was confirmed that, regardless of the experimental group in which apigenin was treated alone or the experimental group in which apigenin-glycated albumin was treated, the expression of IGF-1R, p-AKT, and p-Foxo3a, which induce muscle generation, was increased.

[0186] ​And, for MuRF1 which induces muscle atrophy, it was confirmed that the expression level of MuRF1 which was increased to a very high level by CCM treatment was decreased by apigenin treatment alone and apigenin-glycated albumin treatment, and the MuRF1 expression level of the cell group treated with apigenin-glycated albumin was significantly decreased compared to the MuRF1 expression level of the cell group treated with apigenin alone. It was thus confirmed that apigenin-glycated albumin showed a significant synergistic effect compared to the case of apigenin treatment alone.

[0187] Experimental Example 6: Confirmation of inhibitory effect of anticancer agent on muscle atrophy

[0188] It was confirmed that apigenin-glycated albumin showed an improvement effect on muscle atrophy which can be caused by administration of an anticancer agent.

[0189] First, mouse myocytes C2C12 were cultured, treated with cisplatin 40 μM alone and apigenin at concentrations of 2.5 and 5 μM alone or with apigenin-glycated albumin, and after 48 hours of culture, immunofluorescence staining was performed to confirm the differentiation level into myotube cells, and the results are shown in Figure 12a and Figure 12b

[0190] Referring to Figure 12a and Figure 12b , it was confirmed that cisplatin induced atrophy of myocytes, but in the cell groups treated with apigenin or apigenin-glycated albumin, it was confirmed that the cisplatin-induced atrophy of myocytes was inhibited, and in particular, an excellent atrophy-inhibiting effect was shown in the cell group treated with glycated albumin loaded with apigenin at a concentration of 2.5 μM compared to the cell group treated with apigenin alone at a concentration of 2.5 μM.

[0191] And, as shown in Figure 13 , proteins were extracted from the cells of each experimental group, and Western blot analysis was performed to confirm the expression level of MyoG which induces myogenesis, and as a result, it was confirmed that the decreased MHC and MyoG expression after cisplatin treatment was increased after apigenin treatment, and in particular, in the cell group treated with glycated albumin loaded with apigenin, it was confirmed that the expression level of MyoG which induces myogenesis was significantly increased.

[0192] Thus, it was confirmed that apigenin and glycated albumin play a synergistic role in improving muscle atrophy caused by administration of an anticancer agent.

[0193] Experimental Example 7: Confirmation of in-vivo targeting ability of glycated albumin

[0194] It was confirmed that the injection formulation possibility according to the actual administration route was confirmed using the albumin nanocomplex having a GLUT targeting ability according to the present application. ​

[0195] To evaluate the residual time of the albumin nanocomplex at the injection site and the ability to deliver the drug to the disease site upon direct injection, a muscle injection experiment was performed, and the results are shown in Figure 14

[0196] Referring to Figure 14 , the albumin platform (Albumin(11)) to which 11 ADIBO functional groups were introduced, and the albumin nanocomplex [Glc(6)-Albumin(11), albumin surface ADIBO functional group is 11, and glucose is introduced to 6 thereof] to which glucose was introduced for the purpose of securing GLUT targeting ability in the cell were subjected to muscle injection, and at 0, 3, and 24 hours after the injection, nuclear medicine imaging evaluation was performed using isotopes, and the results showed that a very high amount of residual injection site was exhibited at all times.

[0197] The experimental results are very important in that they can solve the problem that existing drugs are rapidly absorbed after muscle injection, are difficult to sufficiently accumulate at the affected site, and thus cannot exert the intended pharmacological effect, and can set the concentration at which the optimal pharmacological effect is achieved with a small amount of use.

[0198] Experimental Example 8: Evaluation of simultaneous distribution of glycated albumin and apigenin

[0199] Different isotopes were introduced into the albumin and the drug, respectively, and after being simultaneously injected into mice, the kinetics of the drug and the albumin platform were analyzed through real-time imaging.

[0200] A labeling method using the SPECT isotope I-125 (125I-Api) was constructed for apigenin, and a labeling method using the PET isotope Cu-64 (64Cu-Alb) was constructed for albumin, and in order to confirm the targeting ability due to glycosylation, Cu-64 was labeled to the glucose-introduced albumin (64Cu-Glc-Alb).

[0201] After the labeled apigenin (125I-Api), the apigenin and albumin complex (125I-Api / 64Cu-Alb), and the apigenin and glycosylated albumin complex (125I-Api / 64Cu-Glc-Alb) were each subjected to muscle injection, each SPECT and PET images were sequentially obtained, as shown in Figure 15

[0202] Referring to Figure 15 the images, it was confirmed that the apigenin remained at the injection site from the mouse to which the labeled apigenin (125I-Api) was injected, and since the image signal was uniform, it was confirmed that there was no absorption, metabolism, and excretion.

[0203] ​​On the other hand, apigenin and albumin complexes (125I-Api / 64Cu-Alb) and apigenin and glycated albumin complexes (125I-Api / 64Cu-Glc-Alb) were confirmed to be recirculated into the blood via lymphatic vessels after intramuscular injection and then excreted through the intestines.

[0204] This means that in the case of the apigenin / albumin complex, it is absorbed, metabolized, and excreted, thereby increasing the absorption rate of the drug and allowing it to remain in the specific area where it can be effective.

[0205] Furthermore, it exhibited the same morphology as that of PET imaging for up to 8 hours, confirming that it existed in the form of a complex before being absorbed and separated during metabolism and excretion, suggesting the possibility of targeted therapy for specific cells.

[0206] Preparation Example 2: Preparation of ursolic acid-glycated albumin complex

[0207] Ursolic acid was loaded onto the glycated albumin prepared by the method of Preparation Example 1-1.

[0208] 10 equivalents of ursolic acid based on albumin were mixed with albumin, reacted for 1.5 hours, and then purified using a filter to prepare an ursolic acid-glycated albumin complex.

[0209] Experimental Example 9: Confirmation of improvement effect of ursolic acid-glycated albumin on muscle atrophy

[0210] An experiment was conducted to confirm the effect of ursolic acid-glycated albumin on improving muscle atrophy.

[0211] First, mouse muscle cells (C2C12) were cultured. The culture medium was then replaced with cancer cell culture medium (CCM) used to culture mouse colon carcinoma cells (CT26). Muscle atrophy was induced, and cells were treated with 2.5 μM ursolic acid alone or ursolic acid-glycated albumin. After 48 hours of culture, the cells were evaluated for fluorescence expression, thereby confirming the level of myotube differentiation.

[0212] Reference Figure 16a and Figure 16b In cells that had atrophied due to cancer cell culture medium (CCM), MHC expression increased in both the experimental groups treated with ursolic acid alone and the experimental groups treated with ursolic acid-glycated albumin, and it was confirmed that the muscle cells atrophied by CCM were improved.

[0213] In particular, MHC expression was further increased in the cell group treated with ursolic acid-glycated albumin compared to ursolic acid alone, confirming that ursolic acid-glycated albumin exhibited a synergistic effect compared to ursolic acid alone.

[0214] Preparation Example 3: Preparation of soyasapogenol B-glycated albumin complex

[0215] Soyasapogenin B was mixed with 10 equivalents of glycosylated albumin prepared by the method of Preparation Example 1-1, and reacted for 1.5 hours, and then purified using a filter to prepare a soyasapogenin B-glycosylated albumin complex.

[0216] Experimental Example 10: Confirmation of improvement effect of soyasapogenol B-glycated albumin on muscle atrophy

[0217] An experiment for confirming the improvement effect of soyasapogenin B-glycosylated albumin on muscle atrophy was performed.

[0218] First, mouse myoblast C2C12 cells were cultured, and then the medium was replaced with cancer cell medium (CCM) for culturing mouse colon carcinoma cells CT26, and muscle atrophy was induced, and the cells were treated with soyasapogenin B alone at a concentration of 2.5 μM or treated with soyasapogenin B-glycosylated albumin, and after 48 hours of culture, the fluorescence expression of the cells was confirmed to confirm the differentiation level of myotube cells.

[0219] Referring to Figure 17a and Figure 17b In the cells atrophied by the cancer cell medium (CCM), the MHC expression was increased in both the experimental group treated with soyasapogenin B alone and the experimental group treated with soyasapogenin B-glycosylated albumin, and it was confirmed that the muscle cells atrophied by the CCM were improved.

[0220] In particular, it was confirmed that the MHC expression was significantly increased in the cell group treated with soyasapogenin B-glycosylated albumin compared to the group treated with soyasapogenin B alone, and thus it was confirmed that soyasapogenin B-glycosylated albumin showed an excellent synergistic effect compared to the treatment with soyasapogenin B alone.

[0221] Preparation Example 4: Preparation of limonene-glycated albumin complex

[0222] Citronellol was mixed with 10 equivalents of glycosylated albumin prepared by the method of Preparation Example 1-1, and reacted for 1.5 hours, and then purified using a filter to prepare a citronellol-glycosylated albumin complex.

[0223] Experimental Example 11: Confirmation of improvement effect of limonene-glycated albumin on muscle atrophy

[0224] An experiment for confirming the improvement effect of citronellol-glycosylated albumin on muscle atrophy was performed.

[0225] First, mouse myocytes C2C12 were cultured, and then the medium was replaced with cancer cell medium (CCM) in which mouse colon carcinoma cells CT26 were cultured, and muscle atrophy was induced, and cells were treated with limonene alone at a concentration of 2.5 μM or with limonene-sugar albumin, and after 48 hours of culture, fluorescence expression of the cells was confirmed, thereby confirming the differentiation level of myotube cells.

[0226] Reference Figure 18a and Figure 18b In the atrophied cells by the cancer cell medium (CCM), MHC expression was increased in both the experimental group treated with limonene alone and the experimental group treated with limonene-sugar albumin, and it was confirmed that the muscle cells atrophied by the CCM were improved.

[0227] In particular, it was confirmed that MHC expression was significantly increased in the cell group treated with limonene-sugar albumin compared to the group treated with limonene alone, and thus it was confirmed that limonene-sugar albumin showed a synergistic effect compared to the case where limonene alone was treated.

[0228] The above describes some embodiments of the present application, but the present application is not limited to the above-described embodiments, and can be modified and deformed to be implemented within the scope of the gist of the present application. These modifications and deformations should be understood to be also included in the scope of the technical idea of the present application.

Claims

1. An albumin nanocomposite, comprising an albumin bound to an azide or cyclooctyne functional group and a delivery substance bound to an azide or cyclooctyne functional group obtained by a click chemistry reaction, and a phytochemical loaded on the surface of the albumin, characterized in that: The delivery substance comprises a glucose group, When the albumin is bound to the azide functional group, the delivery substance is bound to the cyclooctyne functional group, and when the albumin is bound to the cyclooctyne functional group, the delivery substance is bound to the azide functional group.

2. The albumin nanocomplex according to claim 1, characterized in that The phytochemicals are one or more selected from the group consisting of phenolic compounds and triterpenes.

3. The albumin nanocomplex according to claim 2, characterized in that The phenolic compound is one or more selected from the group consisting of apigenin, citrus alcohol, flavonoids, flavonols, flavanones, flavanols, isoflavones, anthocyanins, stilbenoids, caffeic acid, p-coumaric acid, chrysin, lutein, protoflavonoids, acacetin, luteolin, tangerine, quercetin, kaempferol, genistein, daidzein, naringenin, hesperetin, cyanidin, catechin, curcumin, epigallocatechin gallate and resveratrol.

4. The albumin nanocomplex according to claim 2, characterized in that The triterpenes are one or more selected from the group consisting of ursolic acid, soybean saponin A, soybean saponin B, ursane, oleanane, lupeol, saponin, sterol, oleanolic acid, lupeol, corosolic acid, maslinic acid, betulinic acid, ginsenoside, β-sitosterol, campesterol, stigmasterol, lanosterol and germanium alcohol.

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

6. The albumin nanocomplex according to claim 1, characterized in that The nanocomplex contains 4 to 8 glucose groups.

7. The albumin nanocomplex according to claim 1, characterized in that The sixth carbon atom of the glucose group is combined with an azide or cyclooctyne functional group.

8. The albumin nanocomplex according to claim 1, characterized in that The phytochemicals are loaded with 1 to 20 of the phytochemicals.

9. The albumin nanocomplex according to claim 1, characterized in that The delivery substance further comprises a radioactive isotope, wherein the radioactive isotope is selected from 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 One or more of the group consisting of Lu.

10. The albumin nanocomplex according to claim 9, characterized in that The radioactive isotope is labeled with a chelating agent, and the chelating agent is selected from 1,4,7-triazacyclononane-1,4,7-triacetic acid, 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid, 3-[6,17-dihydroxy-7,10,18,21-tetraoxo-27-[N-acetylhydroxyamino]-6,11,17,22-tetraazaheptacosane]thiourea, diethylenetriaminepentaacetic acid, diaminedithiol, 2-(4'-isothiocyanatobenzyl)-1,4,7-triazacyclononane-1,4,7-triacetic acid, 1,4,7-triazacyclododecane-1,4,7,10-tetraacetic acid, One or more selected from the group consisting of nonane-1-pentanedioic acid-4,7-acetic acid, 2-(4'-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid, 1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid, 2-(4-isothiocyanatobenzyl)-diethylenetriaminepentaacetic acid, 1-(4-isothiocyanatophenyl)-3-[6,17-dihydroxy-7,10,18,21-tetraoxo-27-[N-acetylhydroxyamino]-6,11,17,22-tetraazaheptacosane]thiourea and hydrazinonicotinic acid.

11. The albumin nanocomplex according to claim 1, characterized in that The delivery substance further comprises a fluorescent substance, wherein the fluorescent substance is selected from the group consisting of ferredoxin-NADP+ reductase, cyanine fluorescent substances, tetramethylrhodamine-5-maleimide, Flamma ® One or more selected from the group consisting of a fluorescent substance and indocyanine green.

12. The albumin nanocomplex according to claim 1, characterized in that The nanocomplexes deliver phytochemicals by selectively targeting macrophages overexpressing glucose transporters.

13. A pharmaceutical composition for preventing or treating muscle diseases, characterized in that: Comprising the albumin nanocomplex according to any one of claims 1 to 12.

14. The pharmaceutical composition for preventing or treating muscle diseases according to claim 13, characterized in that: The muscle disease is caused by anticancer agents or inflammatory responses.

15. The pharmaceutical composition for preventing or treating muscle diseases according to claim 14, characterized in that: The anticancer agent is at least one selected from the group consisting of etoposide, doxorubicin, vincristine, cyclophosphamide, ifosfamide, cisplatin, vinorelbine, paclitaxel, docetaxel, gemcitabine, and pemetrexed.

16. The pharmaceutical composition for preventing or treating muscle diseases according to claim 13, characterized in that: The muscle disease is selected from the group consisting of sarcopenia and muscular dystrophy.

17. The pharmaceutical composition for preventing or treating muscle diseases according to claim 13, characterized in that: The pharmaceutical composition inhibits muscle loss, myoblast proliferation, and promotes differentiation into muscle cells.

18. A food composition for preventing or improving muscle diseases, characterized in that: Comprising the albumin nanocomplex according to any one of claims 1 to 12.

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