Zeaxanthin-loaded ferrocene nanoparticles with improved stability, and use thereof

Abstract

The present invention relates to zeaxanthin-loaded ferrocene nanoparticles with improved stability, and a use thereof. More specifically, the present invention relates to zeaxanthin encapsulated with ferrocene nanoparticles, and a use thereof, for example, a use in antioxidants, pharmaceutical compositions, quasi-drug compositions, cosmetic compositions or food compositions.

Description

Zeaxanthin-loaded ferrocene nanoparticles with improved stability and uses thereof

[0001] The present invention relates to zeaxanthin-loaded ferrocene nanoparticles with improved stability and their uses. More specifically, the present invention relates to zeaxanthin encapsulated in ferrocene nanoparticles and their uses, such as in antioxidants, pharmaceutical compositions, quasi-drug compositions, cosmetic compositions, or food compositions.

[0002]

[0003] The exploration of nanoparticle synthesis and its applications has captivated researchers worldwide, making nanotechnology a rapidly growing and innovative field of research. Nanoparticles with sizes ranging from 1 to 100 nm exhibit enhanced properties due to their large surface area and quantum effects. Their applications are diverse, including materials science, environmental research, electronics, energy harvesting, manufacturing, machinery, textiles, and food industries. Novel nanomaterials have attracted increasing attention in recent years due to their utility in biology, engineering, agriculture, and food safety.

[0004] Meanwhile, zeaxanthin is a high-value-added substance belonging to the xanthophyll family of carotenoids, one of approximately 600 natural carotenoids. Zeaxanthin is well known as an antioxidant, eye health supplement, and coloring agent used in livestock and fish farming, and is also known to have anti-inflammatory and anticancer effects. As a free-radical scavenger, zeaxanthin has been confirmed to have a considerably strong ability to remove free oxygen radicals (antioxidant effect) in living tissues.

[0005] [constitutional formula]

[0006]

[0007] Zeaxanthin is concentrated and abundantly distributed around the center of the macula in the human retina. It is known to absorb blue light from strong ultraviolet rays and protect the eye from oxidative stress caused by free radicals. This role contributes to the prevention of age-related eye diseases, such as cataracts and macular degeneration, which can lead to vision loss. Zeaxanthin is also known to be distributed in the heart and skin. Furthermore, zeaxanthin effectively pigments animal fat, skin, and egg yolk, and is therefore attracting attention as a colorant in animal feed to prevent discoloration. Therefore, zeaxanthin can be utilized in diverse industries, including health supplements for eye health, cosmetics, and feed additives, and various products are being developed. However, zeaxanthin's instability and color instability pose limitations when applied to various products, such as reduced efficacy and color discoloration. Consequently, the need to develop products and technologies that address these issues is growing.

[0008] Prior literature

[0009] 1. Republic of Korea Publication Patent No. KR 10-2023-0091552

[0010] The present inventors have made extensive efforts to improve the stability and physicochemical properties of zeaxanthin, and have confirmed that when zeaxanthin is encapsulated in ferrocene nanoparticles, not only is the storage stability improved without precipitation or color change of zeaxanthin occurring, but also significantly improved oxygen radical scavenging activity can be exhibited, thereby completing the present invention.

[0011] However, the problems that the present invention seeks to solve are not limited to the problems mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the description below.

[0012]

[0013] According to the first embodiment, the present invention

[0014] ferrocene-containing polymer; and

[0015] Disclosed are zeaxanthin-loaded ferrocene nanoparticles with improved stability, comprising zeaxanthin loaded within the ferrocene-containing polymer.

[0016] According to one embodiment of the present invention, the zeaxanthin may be contained in an amount of 0.5 to 8 wt%, preferably 0.5 to 4 wt%, and more preferably 1 to 4 wt%, based on the weight of the ferroic acid polymer.

[0017] In one embodiment of the present invention, the ferrocene-containing polymer may include ferrocenylmethyl methacrylate (FMMA), methacrylic acid (MA), and poly(ethylene glycol) methyl ether methacrylate (PEGMA) in a molar ratio of 1:1 to 10:1 to 5.

[0018] According to the second embodiment, the present invention

[0019] Disclosed are an antioxidant, a pharmaceutical composition for preventing, treating or improving a disease caused by excessive production of reactive oxygen species, a cosmetic composition and a food composition comprising zeaxanthin-loaded ferrocene nanoparticles with improved stability according to the first embodiment as an active ingredient.

[0020] According to one embodiment of the present invention, the disease caused by excessive production of active oxygen in the pharmaceutical composition may be stroke, Parkinson's disease, Alzheimer's disease, aging, heart disease, eye disease, ischemia, arteriosclerosis, skin disease, asthma, inflammation, arthritis, autoimmune disease, hyperlipidemia, liver disease, diabetes, cancer, chronic ulcer, burn, or wound.

[0021] According to one embodiment of the present invention, the above-mentioned pharmaceutical composition may be a disinfectant, a shower foam, a gargle, a wet tissue, a detergent soap, a hand wash, a humidifier filler, a mask, an ointment, or a filter filler.

[0022] According to one embodiment of the present invention, the symptoms or diseases caused by active oxygen in the cosmetic composition or food composition may be skin aging, wrinkle formation, skin pigmentation, atopy, acne, psoriasis, or eczema.

[0023] According to one embodiment of the present invention, the cosmetic can be manufactured in the form of an ampoule, cream, lotion, toner, essence or pack.

[0024] According to one embodiment of the present invention, the food may be meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, or vitamin complexes.

[0025]

[0026] Zeaxanthin encapsulated with ferrocene nanoparticles according to the present invention not only has significantly improved storage stability without sedimentation or color change, but also has excellent oxygen radical scavenging activity, is stable, and has excellent biocompatibility, so it is expected to be effectively used in the development of antioxidants, pharmaceutical compositions, quasi-drug compositions, cosmetic compositions, or food compositions.

[0027] Meanwhile, the scope of the present invention is not limited by the effects described above.

[0028]

[0029] Figure 1 shows the results of evaluating the size and dispersion of zeaxanthin-ferrocene nanoparticles according to Example 1.

[0030] Figure 2 shows the results of storage stability evaluation of zeaxanthin-ferrocene nanoparticles according to Example 1.

[0031] Figure 3 shows the results of evaluating the DPPH radical scavenging ability of ZX@FNP2 according to Example 2.

[0032] Figure 4 shows the results of evaluating the ROS production inhibition ability of ZX@FNP2 according to Example 3.

[0033] Figure 5 shows the results of evaluating the size and dispersion of ZX@FNP2 according to Example 4.

[0034] Figure 6 shows the results of storage stability evaluation of ZX@FNP2 according to Example 4.

[0035] Figure 7 shows the results of color stability evaluation of ZX@FNP2 according to Example 4.

[0036]

[0037] Hereinafter, zeaxanthin-loaded ferrocene nanoparticles with improved stability and their uses according to specific embodiments of the invention will be described in detail. However, this is presented as one example of the invention, and the scope of the invention is not limited thereby, and it will be apparent to those skilled in the art that various modifications to the embodiments are possible within the scope of the invention. Unless otherwise specified, throughout this specification, "comprises" or "containing" refers to including a certain component (or component) without any particular limitation, and cannot be interpreted as excluding the addition of other components (or components).

[0038] The term "antioxidant" used in this specification refers to the action of inhibiting oxidation. The human body has a balance of prooxidants and antioxidants, but if this balance becomes unbalanced due to various factors and tilts toward oxidation promotion, oxidative stress is induced, causing potential cell damage and pathological diseases. Reactive oxygen species (ROS), which are the direct cause of this oxidative stress, are unstable and highly reactive, easily react with various biological substances, and attack macromolecules in the body, causing irreversible damage to cells and tissues or leading to mutations, cytotoxicity, and carcinogenesis. NO, HNO2, ONOO - Reactive nitrogen species (RNS) such as these are produced in large quantities by the immune response of macrophages, neutrophils, and other immune cells during an inflammatory response, and ROS are also produced at this time. The above reactive oxygen species oxidize and destroy cells in the body, thereby exposing them to various diseases.

[0039] The term "treatment" as used herein refers to any form of treatment or prevention that provides a benefit to a subject suffering from a disease or at risk of developing a disease, including improving the condition of the subject, delaying the progression of the disease, delaying the onset of symptoms, or slowing the progression of symptoms. Accordingly, the term "treatment" includes prophylactic treatment of a subject that prevents the onset of symptoms. Furthermore, the terms "treatment" and "prevention" are not intended to imply a cure or complete elimination of symptoms.

[0040] The term "improvement" as used herein may mean any action that at least reduces the severity of a symptom, for example, a parameter associated with alleviating or treating a condition.

[0041] 1. Zeaxanthin-loaded ferrocene nanoparticles with improved stability

[0042] The present invention

[0043] Ferrocene-containing polymer; and

[0044] It is an object of the present invention to provide zeaxanthin-loaded ferrocene nanoparticles with improved stability, which contain zeaxanthin loaded inside the ferrocene-containing polymer.

[0045] In the zeaxanthin-loaded ferrocene nanoparticles with improved stability according to the present invention, the zeaxanthin may be contained in an amount of 0.5 to 8 wt%, preferably 0.5 to 4 wt%, and more preferably 1 to 4 wt%, based on the weight of the ferroic acid polymer.

[0046] In the zeaxanthin-loaded ferrocene nanoparticles with improved stability according to the present invention, the ferrocene-containing polymer may include ferrocenylmethyl methacrylate (FMMA), methacrylic acid (MA), and poly(ethylene glycol) methyl ether methacrylate (PEGMA) in a molar ratio of 1:1 to 10:1 to 5.

[0047] In the zeaxanthin-loaded ferrocene nanoparticles with improved stability according to the present invention, the size of the nanoparticles may be 10 to 300 nm, preferably 20 to 200 nm.

[0048] 2. Uses of zeaxanthin-loaded ferrocene nanoparticles with improved stability

[0049] The present invention seeks to provide a use of zeaxanthin-loaded ferrocene nanoparticles with improved stability in an antioxidant, pharmaceutical composition, quasi-drug composition, cosmetic composition or food composition.

[0050] (1) Antioxidant

[0051] The present invention aims to provide an antioxidant comprising zeaxanthin-loaded ferrocene nanoparticles with improved stability as an active ingredient.

[0052] In the antioxidant according to the present invention, the zeaxanthin may be contained in an amount of 0.5 to 8 wt%, preferably 0.5 to 4 wt%, and more preferably 1 to 4 wt%, based on the weight of the ferroic acid polymer.

[0053] In the antioxidant according to the present invention, the ferrocene-containing polymer may include ferrocenylmethyl methacrylate (FMMA), methacrylic acid (MA), and poly(ethylene glycol) methyl ether methacrylate (PEGMA) in a molar ratio of 1:1 to 10:1 to 5.

[0054] In the antioxidant according to the present invention, the size of the nanoparticles may be 10 to 300 nm, preferably 20 to 200 nm.

[0055] (2) Pharmaceutical composition

[0056] The present invention aims to provide a pharmaceutical composition for preventing or treating diseases caused by excessive production of reactive oxygen species, which comprises zeaxanthin-loaded ferrocene nanoparticles with improved stability as an active ingredient.

[0057] In the pharmaceutical composition according to the present invention, the zeaxanthin may be contained in an amount of 0.5 to 8 wt%, preferably 0.5 to 4 wt%, and more preferably 1 to 4 wt%, based on the weight of the ferroic acid polymer.

[0058] In the pharmaceutical composition according to the present invention, the ferrocene-containing polymer may include ferrocenylmethyl methacrylate (FMMA), methacrylic acid (MA), and poly(ethylene glycol) methyl ether methacrylate (PEGMA) in a molar ratio of 1:1 to 10:1 to 5.

[0059] In the pharmaceutical composition according to the present invention, the nanoparticles may be 10 to 300 nm, preferably 20 to 200 nm.

[0060] In the pharmaceutical composition according to the present invention, the disease caused by excessive production of reactive oxygen species may be stroke, Parkinson's disease, Alzheimer's disease, aging, heart disease, eye disease, ischemia, arteriosclerosis, skin disease, asthma, inflammation, arthritis, autoimmune disease, hyperlipidemia, liver disease, eye disease, diabetes, cancer, chronic ulcer, burn, or wound. For example, the above eye diseases include macular degeneration including macular degeneration (e.g., age-related macular degeneration), visual loss, cataracts, glaucoma, ocular peroxidation, retinal damage, dry eye, diabetic retinopathy, macular degeneration including non-exudative age-related macular degeneration and exudative age-related macular degeneration, choroidal neovascularization, retinopathy, acute and chronic macular neuroretinopathy, central serous chorioretinopathy, macular edema, acute multifocal lamellar pigment epitheliopathy, birdshot retinochoroidopathy, posterior scleritis, serpiginous choroiditis, subretinal fibrosis, uveitis syndrome, retinal artery occlusion, central retinal vein occlusion, disseminated intravascular coagulation, branch retinal vein occlusion, hypertensive fundus changes, ocular ischemic syndrome, retinal artery microangiopathy, Coats' disease, foveal telangiectasia, hemilateral retinal vein occlusion, papillophlebitis, central retinal artery The disease may be selected from the group consisting of occlusion, branch retinal artery occlusion, branchial vasculitis, sickle cell retinopathy, angiopattern retinopathy, familial exudative vitreoretinopathy, Eels' disease, proliferative vitreoretinopathy, proliferative diabetic retinopathy, tumor-associated retinal diseases, congenital thickening of the retinal pigment epithelium (RPE), posterior uveal melanoma, choroidal hemangioma, choroidal osteoma, choroidal metastases, combined hamartoma of the retina and retinal pigment epithelium, retinoblastoma, angioproliferative tumor of the fundus, retinal astrocytoma, intraocular lymphoid tumor, myopic retinal degeneration, and acute retinal pigment epitheliitis and glaucoma. The arthritis may be selected from the group consisting of osteoarthritis, degenerative arthritis, rheumatoid arthritis, osteochondritis dissecans, joint ligament damage, meniscus damage, joint malalignment, avascular necrosis, and juvenile idiopathic arthritis.

[0061] The pharmaceutical composition according to the present invention can be administered to a patient by a general oral or parenteral administration method, and can be in any form, solid or liquid. In addition, the pharmaceutical composition according to the present invention can further include one or more pharmaceutically acceptable liquid or solid carriers. In addition, it can be prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. The solid form preparations can be tablets, pills, powders, granules, capsules, pellets, fine granules, or powders, and such solid preparations can be prepared by adding carriers, excipients, and / or diluents to the complex. The carrier, excipient and / or diluent may include lactose, sucrose, dextrose, mannitol, malitol, sorbitol, xylitol, erythritol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, polyvinyl pyrrolidone, magnesium stearate and mineral oil. The liquid form of the preparation may be a solution, suspension or emulsion, and may contain various excipients, such as wetting agents, sweeteners, fragrances and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. Aqueous suspensions suitable for oral use can be prepared by dispersing the finely divided active ingredient in viscous materials such as natural or synthetic gums, resins, methyl cellulose, sodium carboxymethyl cellulose, and known suspending agents.Parenteral medications include injections, drops, solutions, ointments, sprays, suspensions, emulsions, and suppositories. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions can include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases include witepsol, macrogol, cacao butter, laurin, and glycerogelatin.

[0062] (3) Non-medical composition

[0063] The present invention aims to provide a pharmaceutical composition for the prevention or treatment of diseases caused by excessive production of reactive oxygen species, which comprises zeaxanthin-loaded ferrocene nanoparticles with improved stability as an active ingredient.

[0064] In the pharmaceutical composition according to the present invention, the zeaxanthin may be contained in an amount of 0.5 to 8 wt%, preferably 0.5 to 4 wt%, and more preferably 1 to 4 wt%, based on the weight of the ferroic acid polymer.

[0065] In the pharmaceutical composition according to the present invention, the ferrocene-containing polymer may include ferrocenylmethyl methacrylate (FMMA), methacrylic acid (MA), and poly(ethylene glycol) methyl ether methacrylate (PEGMA) in a molar ratio of 1:1 to 10:1 to 5.

[0066] In the pharmaceutical composition according to the present invention, the size of the nanoparticles may be 10 to 300 nm, preferably 20 to 200 nm.

[0067] In the quasi-drug composition according to the present invention, the quasi-drug composition is characterized in that it is a disinfectant, cleanser, shower foam, garglin, wet tissue, detergent soap, hand wash, humidifier filler, mask, ointment, or filter filler.

[0068] (4) Cosmetic composition

[0069] The present invention aims to provide a cosmetic composition for preventing or improving symptoms or diseases caused by excessive production of reactive oxygen species, which comprises zeaxanthin-loaded ferrocene nanoparticles with improved stability as an active ingredient.

[0070] In the cosmetic composition according to the present invention, the symptoms or diseases caused by the active oxygen are characterized as skin aging, wrinkle formation, skin pigmentation, atopy, acne, psoriasis or eczema.

[0071] In the cosmetic composition according to the present invention, the zeaxanthin may be contained in an amount of 0.5 to 8 wt%, preferably 0.5 to 4 wt%, and more preferably 1 to 4 wt%, based on the weight of the ferroic acid polymer.

[0072] In the cosmetic composition according to the present invention, the ferrocene-containing polymer may include ferrocenylmethyl methacrylate (FMMA), methacrylic acid (MA), and poly(ethylene glycol) methyl ether methacrylate (PEGMA) in a molar ratio of 1:1 to 10:1 to 5.

[0073] In the cosmetic composition according to the present invention, the size of the ursodeoxycholic acid nanoparticle complex may be 10 to 300 nm, preferably 20 to 200 nm.

[0074] In the cosmetic composition according to the present invention, the cosmetic may be manufactured in the form of an ampoule, cream, lotion, toner, essence, or pack. In addition, carriers commonly used in formulations, such as dextrin or cyclodextrin, or any other optional additives may be added to facilitate preservation or handling.

[0075] (5) Food composition

[0076] The present invention aims to provide a food composition for preventing or improving symptoms or diseases caused by active oxygen, which comprises zeaxanthin-loaded ferrocene nanoparticles with improved stability as an active ingredient.

[0077] In the food composition according to the present invention, the symptoms or diseases caused by the active oxygen are characterized as skin aging, wrinkle formation, skin pigmentation, atopy, acne, psoriasis or eczema.

[0078] In the food composition according to the present invention, the zeaxanthin may be contained in an amount of 0.5 to 8 wt%, preferably 0.5 to 4 wt%, and more preferably 1 to 4 wt%, based on the weight of the ferroic acid polymer.

[0079] In the food composition according to the present invention, the ferrocene-containing polymer may include ferrocenylmethyl methacrylate (FMMA), methacrylic acid (MA), and poly(ethylene glycol) methyl ether methacrylate (PEGMA) in a molar ratio of 1:1 to 10:1 to 5.

[0080] In the food composition according to the present invention, the size of the nanoparticles may be 10 to 300 nm, preferably 20 to 200 nm.

[0081] In the food composition according to the present invention, the food is characterized in that it is meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, or vitamin complexes.

[0082] In the food composition according to the present invention, the food composition may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, the composition of the present invention may contain fruit pulp for producing natural fruit juice, fruit juice drinks, or vegetable drinks. These ingredients may be used independently or in combination.

[0083]

[0084] Below, various examples are presented to aid understanding of the invention. These examples are provided solely to facilitate understanding of the invention and are not intended to limit the scope of protection of the invention.

[0085] <Example>

[0086] Example 1. Preparation and characterization of zeaxanthin-ferrocene nanoparticles

[0087] 1.1 Preparation of ferrocene polymer

[0088] Three kinds of Poly(FMMA-r-PEGMA-r-MA) polymers were prepared using ferrocenylmethyl methacrylate (FMMA), methacrylic acid (MA), and poly(ethylene glycol) methyl ether methacrylate (PEGMA) as monomers. First, FMMA (0.4 mmol) was added with MA and PEGMA according to the molar ratios shown in Table 1, then azobicisobutyronitrile (AIBN) was added and dissolved in 2 ml of tetrahydrofuran (THF), followed by polymerization at 70°C for 24 hours with magnetic stirring to prepare the polymers.

[0089] PolymerFMMAMAPEGPoly-PC-10.420Poly-PC-20.41.50.5Poly-PC-30.402

[0090] 1.2 Preparation of zeaxanthin-loaded ferrocene nanoparticles The mixed solution containing the prepared ferrocene polymer, zeaxanthin, and ethanol (1 mL) was placed in a 4 mL vial, and the mixed solution was stabilized on a shaking rotor for 2 hours. The amount of zeaxanthin was added at 4 wt% (0.2 mg) based on the weight of the ferrocene polymer. Next, 5 mL of DI water was filled into a 20 mL vial containing a magnetic bar, and the mixed solution was dropped dropwise at a speed of 530 rpm for 10 minutes. After dropwise stirring, the vial was closed and stirred for 60 minutes to stabilize the nanoparticles, and then placed in the dessicator, and ethanol was removed using a cold trap (fixing conditions: -40 degrees, time: 2 h) to obtain zeaxanthin-containing ferrocene nanocapsules. At this time, the zeaxanthin-loaded ferrocene nanoparticles (ZX@FNP) manufactured using the above-mentioned polymers Poly-PC-1, Poly-PC-2 and Poly-PC-3 were named ZX@FNP1, ZX@FNP2 and ZX@FNP3, respectively.

[0091] 1.3 Characteristic Evaluation

[0092] To confirm the stability of the zeaxanthin-loaded ferrocene nanocapsules, size and PDI were measured. As a result, the size and dispersion of ZX@FNP2 and ZX@FNP3 were both found to be approximately 158 nm and less than 0.3, respectively (Fig. 1). The size and dispersion remained constant for up to about 21 days after preparation, and in particular, ZX@FNP2 was found to remain stable even after about 28 days after preparation (Fig. 2).

[0093] Example 2. Evaluation of DPPH radical scavenging ability of ZX@FNP2

[0094] To determine the DPPH radical scavenging activity, DPPH radical scavenging activity was used to measure antioxidant activity, and DPPH radical scavenging activity was measured using a modified method of Blois (1958). 100 μl of 0.2 mM DPPH (2,2-diphenyl-1-picrylhydrazyl, Sigma) solution was dispensed into 100 μl of each sample, left to stand for 30 minutes, and the absorbance was measured at 520 nm using an ELISA reader (Thermo). The control group was measured using the same method by adding a solvent instead of the sample. In addition, in order to correct the absorbance value for the color of the sample itself, ethanol was added instead of 0.2 mM DPPH, and the absorbance was measured. The DPPH radical scavenging activity was calculated using the following mathematical equation 1:

[0095] [Mathematical Formula 1]

[0096] DPPH radical scavenging activity = (1-absorbance of sample added group / absorbance of no-additive group) × 100

[0097] As a result, it was confirmed that the DPPH radical scavenging activity of zeaxanthin itself was approximately 45%, while that of ZX@FNP2 was approximately 60%, showing results comparable to those of the DMSO solution (Fig. 3).

[0098] Example 3. Evaluation of the ROS production inhibition ability of ZX@FNP2

[0099] The cell line used in the test was human keratinocytes HaCaT cell line. 8.0 x 103 cells were seeded per well in a 96-well black plate for fluorescence measurement, and DMEM (Dulbeccos Modification of Eagles Medium, FBS 10%) supplemented with penicillin / streptomycin was used. After culturing for 1 day at 37°C and 5% CO2, the test sample was treated. Serum-free DMEM (FBS free) was used as the medium and cultured for 1 day at 37°C and 5% CO2. After adding the sample and culturing for 24 hours, the remaining medium was removed by washing with HBSS (Hanks' Balanced Salt solution), and 100 μl of DCFH-DA (2',7'-Dichlorofluorescein diacetate, Sigma) prepared at 50 μM in HBSS was added and incubated for 1 hour at 37°C and 5% CO2, and washed with HBSS. After that, 100 μl of HBSS was treated with UVB 20 mJ / cm2. 30 minutes after UV treatment, the fluorescence intensity of DCF (dichlorofluorescein) oxidized to ROS was measured using a fluorescence plate reader (Ex = 485 nm, Em = 530 nm). The measured value was calculated as % Control based on the UV control.

[0100] As a result, ROS increased by about 50% through UV treatment, and ROS production was reduced by about 47% when treated with Trolox, which is a positive control. When treated with ZX extract_D.W (ZX 1 ppm), ROS production was reduced by 15%, and when treated with ZX extract_DMSO (ZX 1 ppm), it was reduced by 28%, whereas when treated with ZX@FNP2 (1 ppm), it was reduced by about 51%, confirming that the ROS scavenging ability of ZX@FNP2 is superior to that of the DMSO solution (Fig. 4).

[0101] Example 4. Preparation and characterization of ZX@FNP2 according to the loading amount of zeaxanthin

[0102] 4.1 Manufacturing of ZX@FNP2

[0103] A mixed solution containing 5 mg of the ferrocene polymer Poly-PC-2, zeaxanthin, and ethanol (1 mL) prepared in the above example was placed in a 4 mL vial, and the mixed solution was stabilized on a shaking rotor for 2 hours. At this time, the amount of zeaxanthin was added in different amounts of 1 wt% (0.05 mg), 2 wt% (0.1 mg), 4 wt% (0.2 mg), and 8 wt% (0.4 mg) based on the weight of the ferrocene polymer. Then, 5 mL of DI water was filled in a 20 mL vial containing a magnetic bar, and the mixed solution was droppedwise at a speed of 530 rpm for 10 minutes. After dropwise, the vial lid was closed and stirred for 60 minutes to stabilize the nanoparticles, and then placed in the dessicator, and ethanol was removed using a cold trap (fixing conditions: -40 degrees, time: 2 h) to obtain zeaxanthin-containing ferrocene nanocapsules.

[0104] 4.2 Characteristic Evaluation

[0105] In order to confirm the stability of the above ZX@FNP2, the size, PDI, and color were measured. As a result, when zeaxanthin was loaded in an amount of 0.5 to 4 wt% based on the weight of the ferrocene polymer, the size and dispersity were found to be about 150 nm or less and less than 0.2, respectively (Fig. 5). The size and dispersity were maintained consistently for about 14 days after manufacture, and in particular, when zeaxanthin was loaded in an amount of 4 wt% based on the weight of the ferrocene polymer, it was confirmed that it was maintained stably even after about 28 days after manufacture (Fig. 6). Meanwhile, when zeaxanthin was loaded in an amount of 8 wt% based on the weight of the ferrocene polymer, it was confirmed that the color was maintained stably without any color change until about 28 days after manufacture, except for a slight increase in turbidity (Fig. 7).

[0106] While specific aspects of the present invention have been described in detail above, it should be apparent to those skilled in the art that these specific descriptions are merely preferred implementation examples and are not intended to limit the scope of the present invention. Therefore, the substantial scope of the present invention is defined by the appended claims and their equivalents.

[0107]

[0108] Zeaxanthin encapsulated with ferrocene nanoparticles according to the present invention not only has significantly improved storage stability without sedimentation or color change, but also has excellent oxygen radical scavenging activity, is stable, and has excellent biocompatibility, so it is expected to be effectively used in the development of antioxidants, pharmaceutical compositions, quasi-drug compositions, cosmetic compositions, or food compositions.

Claims

1. Ferrocene-containing polymer; and Zeaxanthin-loaded ferrocene nanoparticles with improved stability, comprising zeaxanthin loaded into the ferrocene-containing polymer.

2. In paragraph 1, Zeaxanthin-loaded ferrocene nanoparticles with improved stability, characterized in that the zeaxanthin is contained in an amount of 0.5 to 8 wt% based on the weight of the ferroic acid polymer.

3. In paragraph 1, Zeaxanthin-loaded ferrocene nanoparticles with improved stability, characterized in that the ferrocene-containing polymer comprises ferrocenylmethyl methacrylate (FMMA), methacrylic acid (MA), and poly(ethylene glycol) methyl ether methacrylate (PEGMA) in a molar ratio of 1:1 to 10:1 to 5.

4. An antioxidant comprising zeaxanthin-loaded ferrocene nanoparticles with improved stability according to any one of claims 1 to 3 as an active ingredient.

5. A pharmaceutical composition for preventing or treating a disease caused by excessive production of reactive oxygen species, comprising zeaxanthin-loaded ferrocene nanoparticles with improved stability according to any one of claims 1 to 3 as an active ingredient.

6. In paragraph 5, A pharmaceutical composition characterized in that the diseases caused by the excessive production of the above active oxygen are stroke, Parkinson's disease, Alzheimer's disease, aging, heart disease, eye disease, ischemia, arteriosclerosis, skin disease, asthma, inflammation, arthritis, autoimmune disease, hyperlipidemia, liver disease, diabetes, cancer, chronic ulcer, burn or wound.

7. A pharmaceutical composition for the prevention or treatment of a disease caused by excessive production of reactive oxygen species, comprising zeaxanthin-loaded ferrocene nanoparticles with improved stability according to any one of claims 1 to 3 as an active ingredient.

8. In paragraph 7, The above-mentioned pharmaceutical composition is characterized in that it is a disinfectant, a cleansing agent, a shower foam, a garglin, a wet tissue, a detergent soap, a hand wash, a humidifier filler, a mask, an ointment, or a filter filler.

9. A cosmetic composition for preventing or improving symptoms or diseases caused by excessive production of active oxygen, comprising zeaxanthin-loaded ferrocene nanoparticles with improved stability according to any one of claims 1 to 3 as an active ingredient.

10. In paragraph 9, A cosmetic composition characterized in that the symptoms or diseases caused by the above active oxygen are skin aging, wrinkle formation, skin pigmentation, atopy, acne, psoriasis or eczema.

11. In paragraph 9, A cosmetic composition, characterized in that the cosmetic is manufactured in the form of an ampoule, cream, lotion, toner, essence or pack.

12. A food composition for preventing or improving symptoms or diseases caused by active oxygen, comprising zeaxanthin-loaded ferrocene nanoparticles with improved stability according to any one of claims 1 to 3 as an active ingredient.

13. In paragraph 12, A food composition, characterized in that the symptoms or diseases caused by the above active oxygen are skin aging, wrinkle formation, skin pigmentation, atopy, acne, psoriasis or eczema.

14. In paragraph 12, A food composition characterized in that the food is meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages or vitamin complexes.

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