Pharmaceutical composition for inhibiting elastic fibrosis detachment and method for producing said pharmaceutical composition

Abstract

The present invention relates to a pharmaceutical composition for inhibiting the decomposition of elastic fibers, and a method for producing the pharmaceutical composition containing a pomegranate seed extract. [Problem] The purpose of the present invention is to provide a composition having an effect of inhibiting the decomposition of elastic fibers, with the purpose of effectively using waste pomegranate seeds and increasing the added value thereof. [Solution] A pharmaceutical composition for suppressing the decomposition of elastic fibers according to the present invention is characterized by containing a pomegranate seed extract. Furthermore, a preferred embodiment of the pharmaceutical composition for inhibiting the decomposition of elastic fibers according to the present invention is characterized in that the pharmaceutical composition inhibits the decomposition of elastic fibers in fibroblasts of dermis. Furthermore, a preferred embodiment of the pharmaceutical composition for inhibiting the decomposition of elastic fibers according to the present invention is characterized in that the pharmaceutical composition inhibits the activity of an elastin lytic enzyme.

Description

Technical Field

[0001] The present invention relates to a pharmaceutical composition for inhibiting the decomposition of elastic fibers (elastin fibers) and a method for producing the pharmaceutical composition for inhibiting the decomposition of elastic fibers, and in particular to a pharmaceutical composition for inhibiting the decomposition of elastic fibers containing a pomegranate seed extract and a method for producing the pharmaceutical composition for inhibiting the decomposition of elastic fibers containing a pomegranate seed (pomegranate seed) extract. Background Art

[0002] Pomegranate is a fruit in which seeds account for a considerable proportion of the entire fruit, but in principle, the seeds remaining after juicing are discarded. As a few examples of using pomegranate seeds, there is known a nutritional supplement suggestion system using pomegranate seed extract. Regarding the calculation unit of the suggested supplement, when the desired item information is beauty, there is known a nutritional supplement suggestion system that outputs at least one of the following ingredients as the suggested supplement: multivitamins, multiminerals, vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, vitamin E, vitamin P, niacin, biotin, collagen, elastin, polyphenols, pantothenic acid, hyaluronic acid, acetylglucosamine, inositol, coenzyme Q10, α-carotene, β-carotene, soybean saponin, royal jelly, twintose, folic acid, rose petal extract, pomegranate seed extract, black cohosh extract, soybean isoflavones, kudzu isoflavones, squalene, brewer's yeast, fenugreek extract, calcium, magnesium, copper, iron, zinc, and manganese (Patent Document 1). Prior art literature Patent Literature

[0003] Patent Document 1: Japanese Patent Application Laid-Open No. 2011-232989 Summary of the invention Problems to be solved by the invention

[0004] However, in most cases, pomegranate seeds are generally treated as waste, and except for the above-mentioned patent document, pomegranate seeds are not effectively utilized. On the other hand, the effective utilization of waste is being re-examined.

[0005] In recent years, it has been discovered that atopic dermatitis (AD) is caused by loss-of-function mutations in the keratinization-related gene filaggrin (FLG), and the importance of skin barrier function has attracted attention. In this context, if pomegranate seeds, which are waste products, are effective against various diseases, it will be more beneficial.

[0006] Furthermore, in recent years, it has been considered that elastic fibers responsible for skin elasticity among the extracellular matrix proteins constituting the dermis are particularly strongly degraded by ultraviolet irradiation and the like, and thus are greatly involved in wrinkle formation.

[0007] Therefore, it is desired to inhibit the decomposition of elastic fibers by some method.

[0008] Therefore, an object of the present invention is to provide a composition having an effect of inhibiting the decomposition of elastic fibers in order to effectively utilize discarded pomegranate seeds and increase their added value. Means of solving problems

[0009] In order to achieve the above-mentioned object, the present inventors conducted intensive research on the effective utilization of pomegranate seeds, and as a result, found the present invention.

[0010] That is, the pharmaceutical composition for inhibiting the degradation of elastic fibers (elastin fibers) of the present invention is characterized by containing a pomegranate seed extract.

[0011] In addition, a preferred embodiment of the pharmaceutical composition for inhibiting the decomposition of elastic fibers of the present invention is characterized in that the pharmaceutical composition inhibits the decomposition of elastic fibers in dermal fibroblasts.

[0012] In addition, a preferred embodiment of the pharmaceutical composition for inhibiting the degradation of elastic fibers of the present invention is characterized in that the pharmaceutical composition inhibits the activity of an elastin degrading enzyme.

[0013] In addition, a preferred embodiment of the pharmaceutical composition for suppressing the decomposition of elastic fibers of the present invention is characterized in that the decomposition of the elastic fibers is further suppressed under ultraviolet irradiation.

[0014] In a preferred embodiment of the pharmaceutical composition for inhibiting the decomposition of elastic fibers of the present invention, the content of the pomegranate seed extract is 0.1 to 100 μg / mL relative to the total amount of the composition.

[0015] Furthermore, the quasi-drug (quasi-drug) of the present invention is characterized in that it contains the pharmaceutical composition of the present invention as an active ingredient.

[0016] In addition, the method for producing a pharmaceutical composition for inhibiting the decomposition of elastic fibers, characterized in that it contains a pomegranate seed extract, is characterized in that it includes the following steps: a step of immersing a pulverized product obtained by pulverizing pomegranate seeds in a solvent and then separating the supernatant; a step of obtaining a pomegranate seed extract by separating the supernatant; and a step of adjusting the obtained pomegranate seed extract to an effective amount.

[0017] In a preferred embodiment of the pharmaceutical composition for inhibiting decomposition of elastic fibers of the present invention, the solvent is at least one selected from the group consisting of ethyl acetate, ethyl acetate-hexane, ethyl acetate-methanol, ethanol, methanol, hexane and water. Effects of the Invention

[0018] The pharmaceutical composition for inhibiting the decomposition of elastic fibers in the dermis according to the present invention has the advantageous effect of being able to effectively utilize pomegranate seeds that were previously treated as waste. In addition, the pharmaceutical composition for inhibiting the decomposition of elastic fibers according to the present invention has the advantageous effect of inhibiting the decomposition of elastic fibers in the dermis. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 It is a graph showing the evaluation of NEP activity inhibition. (n=5, mean±SE, **p<0.01, ***p<0.001 vs UVA+ control, ###p<0.001 vs UVA- control) Figure 2 The flow chart and yield of fraction preparation are shown. F1 to F4 are obtained by fractionating PSE-EA-H, which has a high yield. DETAILED DESCRIPTION

[0020] The pharmaceutical composition for inhibiting the decomposition of elastic fibers of the present invention is characterized by containing a pomegranate seed extract. The pomegranate seed extract is an extract from pomegranate seeds. The pomegranate seed extract applicable to the present invention is a whole pomegranate seed extract as long as it is derived from pomegranate seeds.

[0021] The target cells are not particularly limited, but the drug effectively acts on skin cells, especially dermal cells. Therefore, the present specification mainly describes dermal cells.

[0022] In addition, a preferred embodiment of the pharmaceutical composition for inhibiting the decomposition of elastic fibers of the present invention is characterized in that the pharmaceutical composition inhibits the decomposition of elastic fibers in dermal fibroblasts.

[0023] In addition, the preferred embodiment of the pharmaceutical composition for suppressing the decomposition of elastic fibers of the present invention is characterized in that the pharmaceutical composition suppresses the activity of elastin decomposing enzyme. In the present invention, as elastin decomposing enzyme, enkephalin (i.e. NEP) can be cited. That is, the pharmaceutical composition can suppress the decomposition of elastic fibers by suppressing the activity of the decomposing enzyme of elastic fibers in the fibroblasts of the dermis, i.e. enkephalin (NEP) etc. That is, the elastic fibers that bear the elasticity of the skin in the extracellular matrix proteins constituting the dermis are subjected to particularly strong decomposition induction due to ultraviolet irradiation, and therefore have a great relationship with wrinkle formation, and as the decomposing enzyme of elastic fibers, enkephalin (NEP) etc. are determined, but surprisingly, in the present invention, by the presence of pomegranate seed extract, the activity of the decomposing enzyme of elastic fibers such as enkephalin (NEP) can be suppressed, and then the decomposition of elastic fibers can be suppressed.

[0024] In the present invention, the dermal fibroblasts are not particularly limited, but from the viewpoint of establishing a cell line, examples thereof include KMST-6 and NHDF (Normal Human Dermal Fibroblasts).

[0025] In addition, a preferred embodiment of the pharmaceutical composition for inhibiting the decomposition of elastic fibers of the present invention is characterized in that the decomposition of the elastic fibers is further inhibited under ultraviolet irradiation. It is well known that elastic fibers in fibroblasts are particularly strongly decomposed by ultraviolet irradiation, and therefore have a great relationship with wrinkle formation, but surprisingly, in the present invention, the activity of elastic fiber decomposing enzymes such as enkephalinase (NEP) can be inhibited under ultraviolet irradiation and non-irradiation. That is, ultraviolet irradiation increases the activity of elastic fiber decomposing enzymes such as enkephalinase (NEP), but as can be seen from the examples described below, in the present invention, the activity of elastic fiber decomposing enzymes is increased by irradiation of ultraviolet A waves (UVA), and the results show that the presence of pomegranate seed extract (PSE) can inhibit the activity of elastic fiber decomposing enzymes.

[0026] In a preferred embodiment of the pharmaceutical composition for inhibiting the degradation of elastic fibers of the present invention, the content of the pomegranate seed extract is 0.1 to 100 μg / mL, more preferably 2 to 50 μg / mL, relative to the total amount of the composition, from the viewpoint of not causing a decrease in cell viability.

[0027] In addition, a preferred embodiment of the pharmaceutical composition for inhibiting the decomposition of elastic fibers of the present invention is characterized in that it includes the following steps: immersing the crushed material obtained by crushing pomegranate seeds in at least one solvent selected from ethyl acetate, ethyl acetate-hexane, ethyl acetate-methanol, ethanol, methanol, water, and hexane, and separating the supernatant to obtain the above-mentioned pomegranate seed extract. For example, shaking extraction can be performed. In the shaking extraction, for example, it can be set on a rotator in a low-temperature chamber of about 4°C, and extraction can be performed while rotating. In addition, a preferred embodiment of the pharmaceutical composition of the present invention is characterized in that the above-mentioned solvent is ethanol.

[0028] In addition, the method for producing a pharmaceutical composition for inhibiting the decomposition of elastic fibers characterized by containing a pomegranate seed extract of the present invention is characterized in that it includes the following steps: a step of immersing a pulverized product obtained by pulverizing pomegranate seeds in a solvent and then separating the supernatant; a step of obtaining a pomegranate seed extract by separating the supernatant; and a step of adjusting the obtained pomegranate seed extract to an effective amount. In addition, a preferred embodiment of the pharmaceutical composition for inhibiting the decomposition of elastic fibers of the present invention is characterized in that the solvent is at least one selected from ethyl acetate, ethyl acetate-hexane, ethyl acetate-methanol, ethanol, methanol, hexane and water.

[0029] In addition, in a preferred embodiment of the method for producing a pharmaceutical composition for inhibiting the decomposition of elastic fibers in the dermis of the present invention, from the perspective of its application and suitability for food, it is characterized in that the ethanol extract of pomegranate seeds extracted with ethanol is further distributed between ethyl acetate and water, and the ethyl acetate layer is concentrated under reduced pressure to obtain a pomegranate seed extract as an ethyl acetate fraction.

[0030] In addition, in a preferred embodiment of the method for producing a pharmaceutical composition for inhibiting the decomposition of elastic fibers in the dermis of the present invention, from the viewpoint of identifying the active ingredient, the ethyl acetate fraction is further partitioned between hexane and methanol to obtain a pomegranate seed extract as an ethyl acetate-hexane fraction and an ethyl acetate-methanol fraction.

[0031] In addition, the ethyl acetate-hexane fraction (PSE-EA-H) and the ethyl acetate-methanol fraction (PSE-EA-Me) are as follows. That is, the pomegranate seed-ethanol extract (PSE) extracted with ethanol is divided into ethyl acetate and water using a separatory funnel, and the solvent of the ethyl acetate layer is concentrated under reduced pressure by an evaporator to prepare the ethyl acetate fraction (PSE-EA), and the solvent of the water layer is freeze-dried to prepare the water fraction (PSE-W). At this time, PSE-EA is further divided into hexane and methanol, and then the solvent is concentrated under reduced pressure by an evaporator, respectively, to obtain the ethyl acetate-hexane fraction (PSE-EA-H) and the ethyl acetate-methanol fraction (PSE-EA-Me).

[0032] Here, first, an example of a method for preparing a pomegranate seed extract is described. First, pomegranate seeds are prepared. The pomegranate seeds are washed and dried as needed. Drying is preferably performed sufficiently. In order to make the subsequent pulverization uniform.

[0033] Next, the pomegranate seeds are crushed. The crushing method is not particularly limited, and a known crusher such as a ball mill, a hammer mill, a roller mill, a rod mill, a sample mill, a pounding mill, a crusher, a mortar, a mixer with a cooling device, etc. can be used. It should be noted that the crushing time can be set to several seconds and repeated more than ten times in consideration of the decomposition of the pomegranate seed composition due to heat generation during crushing.

[0034] Next, the pomegranate seeds are crushed to obtain a crushed product, and then the crushed product is immersed in various solvents. The solvent at this time is not particularly limited, and the solvent can be appropriately set according to the desired effect. In addition, a preferred embodiment of the method for producing a pomegranate seed extract of the present invention is characterized in that the solvent is at least one selected from ethyl acetate, ethyl acetate-hexane, ethyl acetate-methanol, ethanol, methanol, hexane and water. As the solvent, polar and non-polar solvents such as ethyl acetate, ethyl acetate-hexane, ethyl acetate-methanol, ethanol, methanol, water, hexane, ethyl acetate, chloroform, acetone, etc. can be mentioned. Preferably, ethyl acetate, ethyl acetate-hexane, ethyl acetate-methanol, methanol, ethanol, water, etc. can be mentioned.

[0035] The impregnation can be carried out under slow stirring. The above-mentioned pulverized material is impregnated in various solvents to obtain various solutions. Various solutions can be stirred according to the state of the solution, or the solution can be left as it is according to the situation. There is no particular limitation on the stirring time, and the stirring can be continued for 10 hours to 48 hours, preferably for about 1 day (24 hours).

[0036] Then, the pomegranate seed extract can be obtained by fractionating the supernatant. If necessary, the supernatant is evaporated to dryness. The evaporation to dryness can be performed using an evaporator in a warm bath at 20°C to 60°C, preferably 37°C to 40°C. By evaporating to dryness, the pomegranate seed extract can be stored for a long time.

[0037] The components contained in the pomegranate seeds are extracted (extracted) by using solvents with different polarities, and separated by shaking according to their physical properties. Therefore, the types and contents of the components of the pomegranate seed extract differ depending on the solvent used.

[0038] In addition, the quasi-drug (quasi-drug) of the present invention is characterized in that the composition of the present invention is used as an active ingredient. There are no particular limitations on the quasi-drug (quasi-drug) applicable to the present invention as long as the pharmaceutical composition for inhibiting the decomposition of elastic fibers of the present invention is used as an active ingredient.

[0039] <Effective amount> The composition of the present invention is prepared in the form of an effective amount of pomegranate seed extract and a suitable dosage form.

[0040] The dosage of the pomegranate seed extract in the composition of the present invention can be varied according to the condition and severity of the patient, the dosage form, the selected administration route, the number of administrations per day, and the like.

[0041] The dosage of the pomegranate seed extract in the composition of the present invention can be set to 1000 mg / kg / day when administered to mice. In humans, a lower dosage is preferably used depending on the sensitivity of the human body.

[0042] In addition, the dosage form includes oral preparations (tablets, capsules, coated tablets, granules, solutions, syrups), ointments or creams for application, etc. From the perspective of skin moisture replenishment and barrier function, the subject of administration can be female, male, adult, child, etc.

[0043] The dosage form may contain other conventional ingredients such as a storage stabilizer, a sweetener, a colorant, a flavoring agent, and the like.

[0044] <Acute toxicity test> Regarding the ingredients contained in pomegranate seed extract, since it contains linolenic acid, it is considered to be non-toxic.

[0045] The abbreviations are as follows. FBS: Fetal bovine serum F1: PSE-ethyl acetate-hexane-fraction 1 F2: PSE-ethyl acetate-hexane-fraction 2 F3: PSE-ethyl acetate-hexane-fraction 3 F4: PSE-ethyl acetate-hexane-fraction 4 HA: Hyaluronic acid HABP: Hyaluronan Binding Protein HAS: Hyaluronic acid synthase PBS: Phosphate buffered saline PSE: Punica gtanatumSeed Ethanol extract ·PSE-Bu-Me: PSE-butanol-methanol fraction ·PSE-EA: PSE-ethyl acetate fraction PSE-EA-H: PSE-ethyl acetate-hexane fraction ·PSE-EA-Me: PSE-ethyl acetate-methanol fraction ·PSE-W: PSE-water fraction UV: Ultraviolet light Example

[0046] An embodiment of the present invention will be described below, but the present invention is not limited to the following embodiment.

[0047] Example 1 First, regarding the skin functionality of pomegranate extract, the evaluation of elastase activity was specifically investigated.

[0048] <Abbreviation list> ·PSE: Punica granatum Seed Ethanol extract ·FBS: Fetal Bovine Serum

[0049] <Research summary> Among the extracellular matrix proteins that make up the dermis, elastic fibers, which are responsible for skin elasticity, are particularly strongly induced to decompose by ultraviolet irradiation, and thus are highly related to wrinkle formation. Neprilysin (NEP) has been identified as the degrading enzyme of elastic fibers. In this experiment, an evaluation was carried out on whether the NEP activity in the crude enzyme solution prepared from dermal fibroblasts irradiated / non-irradiated with ultraviolet A (UVA) was inhibited in the presence of PSE, through the inhibition evaluation of elastase activity by an enzyme-substrate reaction.

[0050] First, ethanol was used as a solvent to attempt the preparation of pomegranate seed extract. The pomegranate seeds were crushed and extracted by shaking with 100% ethanol. It was concentrated to dryness and redissolved to prepare the pomegranate seed-ethanol extract for the experiment. Further, the pomegranate seed-ethanol extract (hereinafter also referred to as PSE) extracted with ethanol was partitioned (extracted) between ethyl acetate and water using a separating funnel. The solvent in the ethyl acetate layer was concentrated under reduced pressure using an evaporator to prepare an ethyl acetate fraction (PSE-EA). The aqueous layer was lyophilized to prepare an aqueous fraction (PSE-W). At this time, PSE-EA was partitioned between hexane and methanol, and then the solvents were concentrated under reduced pressure using an evaporator, and the resulting fractions became an ethyl acetate-hexane fraction (hereinafter also referred to as PSE-EA-H) and an ethyl acetate-methanol fraction (hereinafter also referred to as PSE-EA-Me). Specifically, it is described as follows.

[0051] <Preparation method of PSE (pomegranate seed-ethanol extract)> The dried pomegranate seeds were powdered using a grinder, and 10 times the amount of 100% ethanol (manufactured by Wako Pure Chemical Industries, Ltd.) was added, and low-temperature shaking extraction was carried out at 4°C for 24 hours. After 24 hours, centrifugation was carried out at room temperature at 3000 rpm for 20 minutes, and the supernatant was recovered. The concentrate obtained by concentrating the supernatant under reduced pressure was used as the pomegranate seed-ethanol extract (PSE: Punica granatum Seed Ethanol extract).

[0052] <Preparation method of ethyl acetate-hexane fraction (PSE-EA-H) and ethyl acetate-methanol fraction (PSE-EA-Me)> 2.00 g of PSE was distributed in ethyl acetate (100 ml×3) and Mil iQ water (ultrapure water) (100 ml) using a separatory funnel, and the solvent of the ethyl acetate layer was concentrated under reduced pressure by an evaporator to prepare an ethyl acetate fraction (PSE-EA). The solvent of the water layer was freeze-dried to prepare a water fraction (PSE-W). PSE-EA was further distributed in hexane (100 ml×3) and methanol (100 ml), and the solvent was then concentrated under reduced pressure by an evaporator to obtain an ethyl acetate-hexane fraction (PSE-EA-H) and an ethyl acetate-methanol fraction (PSE-EA-Me). The high-yield PSE-EA-H was distributed through a chromatographic column (30φ×300mm) in the order of hexane (100ml), a mixture of hexane and ethyl acetate (hexane:ethyl acetate = 9:1 (hexane 90mL, ethyl acetate 10mL), hexane:ethyl acetate = 8:2 (hexane 80mL, ethyl acetate 20mL), hexane:ethyl acetate = 6:4 (hexane 40mL, ethyl acetate 60mL)), ethyl acetate (100ml), and methanol (100ml). The fraction yield was calculated according to the following formula: Figure 2 A flow chart showing fraction preparation is shown.

[0053] Yield (%) = (mass of fraction (g) / mass of PSE (g)) × 100

[0054] The prepared extracts were stored frozen at -20 °C until used in experiments.

[0055] <Experimental methods> <Cultivation and UVA irradiation> KMST-6 cells were cultured at a final concentration of 7.0 × 10 4 Cells / mL were inoculated into a 10 cm dish. Pre-cultured with 10% FBS MEMα for about 15 hours to allow cells to fully adhere. After washing the dish once with PBS, the culture medium was replaced with MEMα containing 5% FBS but without phenol red, and irradiated on ice at 10 J / cm 2 Considering the increase in liquid temperature caused by UVA irradiation, the UVA non-irradiation group was kept on ice at 32°C until the treatment of the UVA irradiation group was completed. The culture dish was then returned to the incubator and cultured for 24 hours.

[0056] <Preparation of crude enzyme solution> After washing the culture dish three times with PBS, use 1000μL chip to recover cells and centrifuge at 1200rpm for 5 minutes to make cell blocks. Extract the supernatant, add 0.1% Triton-X-100 (non-fluorescent) / 0.2M Tris-HCl (pH8.0) to the supernatant, and ultrasonicate on ice for 1 minute to prepare cell disruption solution. Centrifuge at 8000×g for 20 minutes and use the supernatant as the crude enzyme solution for fibroblasts.

[0057] <Evaluation of NEP activity inhibition> The crude enzyme solution of fibroblasts diluted to a protein concentration of 60 μg / mL was added to a 96-well black plate at 50 μL / well together with 50 μL of PSE (or control) / MES buffer solution adjusted to a concentration twice the target, and gently shaken for pre-incubation. After 15 minutes, 2 μL of 10 mM glutaryl-Ala-Ala-Phe-4-methoxy-β-naphthamide (NEP substrate) was added, and the enzyme substrate decomposition reaction was carried out at 37°C for 1 hour. 2 μL of 100 μM phosphonyl dipeptide was added to stop the reaction, and then 2 μL of 25 U / mL leucine aminopeptidase M was added and incubated for 1 hour to separate the fluorophore from the substrate decomposition product. The fluorescence intensity at an excitation wavelength of 340 nm and a fluorescence wavelength of 425 nm was calculated as the substrate decomposition rate = NEP activity, and compared with the activity of the control group without pomegranate seed extract.

[0058] <Experimental Results> Comparing the UVA irradiation / non-irradiation controls, the NEP activity in the crude enzyme solution prepared by UVA irradiated cells was 30% higher than that in the non-irradiated group. Concentration-dependent inhibition of NEP activity was observed by adding PSE, and inhibition was also confirmed for the state where NEP activity was increased by UVA irradiation. When 50μg / mL of PSE was added, the NEP activity of the crude enzyme solution prepared by UVA irradiated cells (compared to the non-irradiated control) was inhibited from about 130% to 104%. The above results show that PSE inhibits the decomposition of elastic fibers caused by NEP.

[0059] in addition, Figure 2 The flow chart and yield of fraction preparation are shown. F1 to F4 are obtained by distributing PSE-EA-H with a large yield. It is explained that further fractionation of PSE-EA-H to obtain F1 to F4 means that the fractions obtained after passing PSE-EA-H through the chromatographic column according to F1 being a mixture of hexane and ethyl acetate in a ratio of 8:2, F2 being a mixture of hexane and ethyl acetate in a ratio of 6:4, F3 being 100% ethyl acetate, and F4 being 100% methanol. The same effect was obtained with these pomegranate seed extracts. Industrial Applicability

[0060] According to the present invention, it is not enough to mention the contribution to the medical industry field, the social contribution such as the effective utilization of waste is high, and the utilization value is high in a wide range.

Claims

1. A pharmaceutical composition for inhibiting the decomposition of elastic fibers, comprising a pomegranate seed extract.

2. The pharmaceutical composition for inhibiting the decomposition of elastic fibers according to claim 1, characterized in that: The pharmaceutical composition inhibits the breakdown of elastic fibers in fibroblasts of the dermis.

3. The pharmaceutical composition for inhibiting the decomposition of elastic fibers according to claim 1 or 2, characterized in that: The pharmaceutical composition inhibits the activity of elastin degrading enzymes.

4. The pharmaceutical composition for inhibiting the decomposition of elastic fibers according to any one of claims 1 to 3, characterized in that: Furthermore, the decomposition of the elastic fiber is suppressed under ultraviolet irradiation.

5. The pharmaceutical composition for inhibiting the decomposition of elastic fibers according to any one of claims 1 to 4, characterized in that: The content of the pomegranate seed extract is 0.1-100 μg / mL relative to the total amount of the composition.

6. A quasi-drug, characterized in that The pharmaceutical composition according to any one of claims 1 to 5 is used as an active ingredient.

7. A method for producing a pharmaceutical composition for inhibiting the decomposition of elastic fibers containing a pomegranate seed extract, characterized in that: Including the following processes: The process of immersing the crushed product obtained by crushing pomegranate seeds in a solvent and then collecting the supernatant. a step of obtaining a pomegranate seed extract by fractionating the supernatant, and A step of adjusting the obtained pomegranate seed extract to an effective amount.

8. The method according to claim 7, characterized in that The solvent is at least one selected from the group consisting of ethyl acetate, ethyl acetate-hexane, ethyl acetate-methanol, ethanol, methanol, hexane and water.

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