Pharmaceutical composition for suppressing elastin fiber decomposition, and pomegranate seed extract-containing pharmaceutical composition for suppressing elastin fiber decomposition

Abstract

To provide pharmaceutical compositions for suppressing elastin fiber decomposition to achieve effective utilization of and value addition to disposed pomegranate seeds.SOLUTION: The pharmaceutical compositions for suppressing elastin fiber decomposition of the invention comprise a pomegranate seed extract. In a preferred embodiment of the pharmaceutical compositions for suppressing elastin fiber decomposition, the pharmaceutical composition suppresses elastin fiber decomposition in the dermal fibroblasts. In a preferred embodiment of the pharmaceutical compositions for suppressing elastin fiber decomposition, the pharmaceutical composition inhibits activities of elastin-degrading enzymes.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a pharmaceutical composition for inhibiting the degradation of elastin fibers and a method for producing a pharmaceutical composition for inhibiting the degradation of elastin fibers, and in particular to a pharmaceutical composition for inhibiting the degradation of elastin fibers that contains pomegranate seed extract, and a method for producing a pharmaceutical composition for inhibiting the degradation of elastin fibers. [Background technology]

[0002] Pomegranate is a fruit in which seeds account for a significant proportion of the entire fruit, but as a rule, the seeds remaining after squeezing are discarded. A few examples of using pomegranate seeds include a supplement suggestion system using pomegranate seed extract, in which a suggested supplement calculation means outputs at least one of the following as suggested supplements when the desired item information is beauty: 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, alpha carotene, beta carotene, soybean saponin, royal jelly, twin toes, folic acid, rose petal extract, pomegranate seed extract, black cohosh extract, soybean isoflavones, kudzu isoflavone squalene, brewer's yeast, fenugreek extract, calcium, magnesium, copper, iron, zinc, and manganese (Patent Document 1). [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Patent Publication No. 2011-232989 DISCLOSURE OF THEINVENTION [Problem to be solved by the invention]

[0004] However, in most cases, pomegranate seeds are currently disposed of as waste, and are not effectively utilized except for those described in the above patent documents. On the other hand, effective utilization of waste materials is being reconsidered.

[0005] In recent years, it has been discovered that atopic dermatitis (AD) is caused by a loss-of-function mutation in the keratinization-related gene filaggrin (FLG), and the importance of skin barrier function has been drawing attention. Under these circumstances, it would be even more beneficial if pomegranate seeds, a waste product, were effective against various diseases.

[0006] Furthermore, in recent years, it has been pointed out that elastin fibers, which are the extracellular matrix proteins that make up the dermis and are responsible for the elasticity of the skin, are particularly susceptible to intense decomposition induction by exposure to ultraviolet rays and the like, and are therefore closely related to the formation of wrinkles.

[0007] Therefore, it is desirable to have some method to inhibit the degradation of elastin fibers.

[0008] Therefore, an object of the present invention is to provide a composition that has the effect of inhibiting the decomposition of elastin fibers, with the aim of effectively utilizing and adding value to discarded pomegranate seeds. [Means for solving the problem]

[0009] In order to achieve the above object, the present inventors conducted extensive research into effective utilization of pomegranate seeds and came up with the present invention.

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

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

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

[0013] In a preferred embodiment of the pharmaceutical composition for inhibiting the degradation of elastin fibers of the present invention, the composition is further characterized in that it inhibits the degradation of elastin fibers under ultraviolet light irradiation.

[0014] In a preferred embodiment of the pharmaceutical composition for inhibiting the degradation of elastin fibers according to 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 of the present invention is characterized by containing the pharmaceutical composition of the present invention as an active ingredient.

[0016] In addition, the manufacturing method of the pharmaceutical composition for inhibiting the degradation of elastin fibers of the present invention is characterized by containing a pomegranate seed extract, which comprises the steps of crushing pomegranate seeds, immersing the crushed product in a solvent, and then separating the supernatant, obtaining a pomegranate seed extract by separating the supernatant, and adjusting the obtained pomegranate seed extract to an effective amount.

[0017] Furthermore, in a preferred embodiment of the pharmaceutical composition for inhibiting the degradation of elastin 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. Effect of the Invention

[0018] The pharmaceutical composition for inhibiting the degradation of elastin fibers in the dermis of the present invention has the advantageous effect of making effective use of pomegranate seeds, which have traditionally been treated as waste. In addition, the pharmaceutical composition for inhibiting the degradation of elastin fibers in the dermis of the present invention has the advantageous effect of inhibiting the degradation of elastin fibers in the dermis. [Brief description of the drawings]

[0019] [Figure 1] Figure 1 shows the evaluation of NEP activity inhibition. (n=5, mean ± standard error, **p < 0.01, ***p < 0.001 vs. UVA+ Control, ###p < 0.001 vs. UVA- Control) [Diagram 2] Figure 2 shows the flow chart of fraction preparation and the yields. F1 to F4 were obtained by distributing PSE-EA-H, which had a high yield. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0020] The pharmaceutical composition for inhibiting the degradation of elastin fibers according to the present invention is characterized by containing pomegranate seed extract. The pomegranate seed extract is an extract derived from pomegranate seeds. The pomegranate seed extract applied to the present invention covers all pomegranate seed extracts as long as they are derived from pomegranate seeds.

[0021] Although the target cells are not particularly limited, the cells on which the present invention acts are particularly effective are skin cells, particularly dermal cells, and therefore, the present specification will mainly describe dermal cells.

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

[0023] In a preferred embodiment of the pharmaceutical composition for inhibiting the degradation of elastin fibers of the present invention, the pharmaceutical composition is characterized by inhibiting the activity of elastin degrading enzymes. In the present invention, an example of the elastin degrading enzyme is Neprilysin (NEP). That is, the pharmaceutical composition can inhibit the degradation of elastin fibers by inhibiting the activity of elastin fiber degrading enzymes, i.e., Neprilysin (NEP) and the like, in dermal fibroblasts. That is, elastin fibers, which are responsible for the elasticity of the skin among the extracellular matrix proteins constituting the dermis, are particularly strongly degraded by ultraviolet irradiation and are therefore closely involved in the formation of wrinkles, and Neprilysin (NEP) and the like have been identified as elastin fiber degrading enzymes. Surprisingly, in the present invention, it has been found that the presence of pomegranate seed extract makes it possible to inhibit the activity of elastin fiber degrading enzymes such as Neprilysin (NEP), and thus inhibit the degradation of elastin fibers.

[0024] In the present invention, the dermal fibroblasts are not particularly limited, but examples thereof include KMST-6, NHDF (Normal Human Dermal Fibroblasts), etc., which are established cell lines.

[0025] In addition, in a preferred embodiment of the pharmaceutical composition for inhibiting the degradation of elastin fiber of the present invention, the composition is further characterized by inhibiting the degradation of the elastin fiber under ultraviolet irradiation.It is known that elastin fiber in fibroblasts is strongly decomposed by ultraviolet irradiation and is therefore closely related to wrinkle formation, but surprisingly, it has been found that the activity of elastin fiber decomposition enzymes such as Neprilysin (NEP) can be inhibited under both ultraviolet irradiation and non-irradiation in the present invention.That is, it is said that ultraviolet irradiation increases the activity of elastin fiber decomposition enzymes such as Neprilysin (NEP), but as is clear from the examples described below, it has been found that the activity of elastin fiber decomposition enzymes can be inhibited by the presence of pomegranate seed extract (PSE) in the present invention, where the activity of elastin fiber decomposition enzymes increases due to irradiation with ultraviolet A rays (UVA).

[0026] Furthermore, in a preferred embodiment of the pharmaceutical composition for inhibiting the degradation of elastin 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 a preferred embodiment of the pharmaceutical composition for inhibiting the degradation of elastin fibers of the present invention, the pomegranate seed extract is obtained by pulverizing pomegranate seeds, immersing the pulverized product in at least one solvent selected from the group consisting of ethyl acetate, ethyl acetate-hexane, ethyl acetate-methanol, ethanol, methanol, water, and hexane, and separating the supernatant. For example, shaking extraction can be performed. In shaking extraction, for example, extraction can be performed while rotating the product by setting it on a rotator in a low-temperature room at about 4°C. In a preferred embodiment of the pharmaceutical composition of the present invention, the solvent is ethanol.

[0028] In addition, the method for producing a pharmaceutical composition for inhibiting the degradation of elastin fibers of the present invention is characterized by comprising the steps of: immersing the pulverized product obtained by pulverizing pomegranate seeds in a solvent and then separating the supernatant; obtaining a pomegranate seed extract by separating the supernatant; and adjusting the obtained pomegranate seed extract to an effective amount. In a preferred embodiment of the pharmaceutical composition for inhibiting the degradation of elastin 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.

[0029] Furthermore, in a preferred embodiment of the method for producing a pharmaceutical composition for inhibiting the degradation of elastin fibers in the dermis of the present invention, from the viewpoint of anticipated application to food, the method is characterized in that the pomegranate seed ethanol extract extracted with ethanol is further partitioned with ethyl acetate and water, and the ethyl acetate layer is concentrated under reduced pressure to obtain 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 degradation of elastin fibers in the dermis of the present invention, from the viewpoint of identifying the active ingredient, the method is further characterized in that the ethyl acetate fraction is partitioned with hexane and methanol to obtain pomegranate seed extract as an ethyl acetate-hexane fraction and an ethyl acetate-methanol fraction.

[0031] The ethyl acetate-hexane fraction (PSE-EA-H) and the ethyl acetate-methanol fraction (PSE-EA-Me) are as follows: That is, when pomegranate seed ethanol extract (PSE) extracted with ethanol is partitioned between ethyl acetate and water using a separatory funnel, and the solvent of the ethyl acetate layer is concentrated under reduced pressure using an evaporator to obtain the ethyl acetate fraction (PSE-EA), and the solvent of the water layer is freeze-dried to obtain the water fraction (PSE-W), PSE-EA is partitioned between hexane and methanol, and the solvents are then each concentrated under reduced pressure using an evaporator to obtain the resulting fractions, which can be called the ethyl acetate-hexane fraction (PSE-EA-H) and the ethyl acetate-methanol fraction (PSE-EA-Me).

[0032] Here, an example of a method for preparing pomegranate seed extract will be described. First, pomegranate seeds are prepared. The pomegranate seeds are washed and dried as necessary. It is preferable to dry the seeds thoroughly so that the subsequent grinding can be performed uniformly.

[0033] Next, the pomegranate seeds are pulverized. The pulverization method is not particularly limited, and known pulverizers such as ball mills, hammer mills, roller mills, rod mills, sample mills, stamp mills, disintegrators, mortars, and blenders with cooling devices can be used. Note that, since heat generated during pulverization may cause decomposition of the pomegranate seed composition, the pulverization time is set to several seconds, and the pulverization can be repeated a dozen times.

[0034] Next, the pomegranate seeds are crushed to obtain a crushed product, and the crushed product is immersed in various solvents. In this case, the solvent is not particularly limited, and can be appropriately selected according to the desired effect. In a preferred embodiment of the method for producing a pomegranate seed extract 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. Examples of the solvent include ethyl acetate, ethyl acetate-hexane, ethyl acetate-methanol, ethanol, methanol, water, hexane, ethyl acetate, chloroform, acetone, and other polar and non-polar solvents. Preferred examples include ethyl acetate, ethyl acetate-hexane, ethyl acetate-methanol, methanol, ethanol, and water.

[0035] The immersion can be carried out under gentle stirring. The pulverized material is immersed in various solvents to obtain various solutions. The various solutions are stirred depending on the state of the solution, and in some cases the solution may be left as is. When stirring, the stirring can be continued for 10 hours to 48 hours, preferably for about one day (24 hours), although there is no particular limitation.

[0036] Thereafter, the supernatant is separated to obtain the pomegranate seed extract. If necessary, the supernatant is evaporated to dryness. The evaporation to dryness can be carried out using an evaporator on 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 period of time.

[0037] The components contained in pomegranate seeds are extracted with solvents of different polarity and are divided according to their physical properties. Therefore, the type and content of the components in the pomegranate seed extract varies depending on the solvent used.

[0038] Furthermore, the quasi-drug of the present invention is characterized by containing the composition of the present invention as an active ingredient. There are no particular limitations on the quasi-drug applicable to the present invention, so long as it contains the pharmaceutical composition for inhibiting the degradation of elastin fibers of the present invention as an active ingredient.

[0039] <Effective amount> The compositions according to the present invention are prepared by providing an effective amount of pomegranate seed extract and a suitable dosage form.

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

[0041] The dosage of pomegranate seed extract in the composition of the present invention can be set at 1000 mg / kg / day when tested in mice, and a lower dosage is preferable in humans due to differences in sensitivity, etc.

[0042] The dosage form may be an oral agent (tablet, capsule, coated tablet, granule, solution, syrup), or an ointment or cream for application. From the viewpoint of skin hydration and barrier function, the subject of administration may be any patient, regardless of gender, gender, adult, or child.

[0043] The dosage form may contain other conventional ingredients, such as preservatives, sweeteners, colorants, flavoring agents, and the like.

[0044] <Acute toxicity test> As for the ingredients contained in pomegranate seed extract, including linoleic acid, it is believed that there is no toxicity.

[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:Hyaluronic Acid Binding Protein ·HAS:Hyaluronic Acid Synthase ·PBS: Phosphate Buffered Saline · PSE: Punica granatum Seed 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 EXAMPLES

[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, the skin functionality of pomegranate extract was examined, specifically, its elastase activity was evaluated.

[0048] <List of abbreviations> · PSE: Punica granatum Seed Ethanol extract ·FBS: Fetal Bovine Serum

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

[0050] First, an attempt was made to prepare pomegranate seed extract using ethanol as a solvent. Pomegranate seeds were crushed and shaken and extracted with 100% ethanol. These were concentrated to dryness and redissolved, and used in the experiment as pomegranate seed ethanol extract. Furthermore, the pomegranate seed ethanol extract extracted with ethanol (hereinafter also referred to as PSE) was partitioned with ethyl acetate and water using a separating funnel, the solvent in the ethyl acetate layer was concentrated under reduced pressure using an evaporator, and when the aqueous layer was freeze-dried to obtain a water fraction (PSE-W), after partitioning PSE-EA with hexane and methanol, the solvents were each concentrated under reduced pressure using an evaporator, and the obtained fractions were used as 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 as follows.

[0051] <Preparation method of PSE (Punica granatum Seed Ethanol extract)> Dried pomegranate seeds were powdered with a miller, 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 3,000 rpm for 20 minutes, and the supernatant was collected. The concentrate obtained by concentrating the supernatant under reduced pressure was used as 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)> 2.00g of PSE was partitioned between ethyl acetate (100ml x 3) and MiliQ water (100ml) using a separatory funnel, and the solvent in the ethyl acetate layer was concentrated under reduced pressure using an evaporator to obtain the ethyl acetate fraction (PSE-EA). The solvent in the water layer was freeze-dried to obtain the water fraction (PSE-W). PSE-EA was further partitioned between hexane (100ml x 3) and methanol (100ml), and the solvents were then each concentrated under reduced pressure using an evaporator to obtain the ethyl acetate-hexane fraction (PSE-EA-H) and the ethyl acetate-methanol fraction (PSE-EA-Me). The PSE-EA-H with the highest yield was passed through a column (30φ×300mm) in the following order: 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) were poured and distributed. The fraction yield was calculated from the following formula, and the flow chart of fraction preparation is shown in Figure 2.

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

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

[0055] <Experimental Method> <Culture and UVA irradiation> KMST-6 cells were cultured in a 10 cm dish at a final concentration of 7.0 x 10 4 The cells were pre-cultured in 10% FBS MEMα for approximately 15 hours to allow sufficient cell adhesion. After washing the dish once with PBS, the culture medium was replaced with MEMα containing 5% FBS and no phenol red, and the cells were incubated on ice for 10 J / cm 2It was irradiated with UVA. Considering the increase in liquid temperature due to UVA irradiation, the non-UVA-irradiated group was at 32°C and left standing on ice until the treatment of the UVA-irradiated group was completed. Then, the dish was returned to the incubator and cultured for 24 hours.

[0056] <Preparation of crude enzyme solution> After washing the dish with PBS three times, the cells were collected using a 1000 μL pipette tip and centrifuged at 1,200 rpm for 5 minutes to prepare cell pellets. The supernatant was aspirated, 0.1% Triton-X-100 (non-fluorescent) / 0.2 M Tris-HCl (pH 8.0) was added, and ultrasonic treatment was performed on ice for 1 minute to prepare a cell lysate. It was centrifuged at 8,000×g for 20 minutes, and the supernatant was used as the fibroblast crude enzyme solution.

[0057] <Evaluation of NEP activity inhibition> The fibroblast crude enzyme solution was diluted so that the protein concentration became 60 μg / mL, and 50 μL / well was added to a 96-well black plate together with 50 μL of a PSE (or Control) / MES Buffer solution prepared at twice the target concentration, and gently shaken for pre-incubation. After 15 minutes, 2 μL of 10 mM Glutaryl-Ala-Ala-Phe-4-methoxy-β-naphthylamide (NEP substrate) was added, and an enzyme-substrate degradation reaction was carried out at 37°C for 1 hour. After adding 2 μL of 100 μM Phosphoramidon to stop the reaction, 2 μL of 25 U / mL Leucine aminopeptidase M was added and incubated for 1 hour to separate the fluorophore from the degradation product of the substrate. The fluorescence intensity at an excitation wavelength of 340 nm and an emission wavelength of 425 nm was calculated as the substrate degradation rate = NEP activity and compared with the activity of Control in the absence of the pomegranate seed extract.

[0058] <Experimental results> When comparing UVA-irradiated and non-irradiated controls, the NEP activity in the crude enzyme solution prepared from UVA-irradiated cells was 30% higher than that of the non-irradiated one. The addition of PSE resulted in a concentration-dependent inhibition of NEP activity, and similar inhibition was confirmed even when NEP activity was increased by UVA irradiation. The addition of 50μg / mL PSE reduced the NEP activity of the crude enzyme solution prepared from UVA-irradiated cells from approximately 130% (compared to the non-irradiated control) to 104%. These results suggest that PSE inhibits the degradation of elastin fibers by NEP.

[0059] Figure 2 shows the flow chart of fraction preparation and the yield. F1 to F4 were obtained by partitioning PSE-EA-H, which had a high yield. F1 to F4 were obtained by further fractionating PSE-EA-H, meaning that F1 was obtained after running the column with a mixture of 8:2 hexane and ethyl acetate, F2 was obtained with a mixture of 6:4 hexane and ethyl acetate, F3 was obtained with 100% ethyl acetate, and F4 was obtained with 100% methanol. The same effect was obtained with these pomegranate seed extracts. [Industrial Applicability]

[0060] The present invention not only contributes to the medical industry, but also contributes greatly to society through the effective use of waste materials, and has a wide range of useful value.

Claims

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

2. The pharmaceutical composition for inhibiting the degradation of elastin fibers according to claim 1, characterized in that the pharmaceutical composition inhibits the degradation of elastin fibers in dermal fibroblasts.

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

4. A pharmaceutical composition for inhibiting the degradation of elastin fibers according to any one of claims 1 to 3, further comprising: a composition for inhibiting the degradation of elastin fibers under ultraviolet light irradiation.

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

6. A quasi-drug comprising the pharmaceutical composition according to any one of claims 1 to 5 as an active ingredient.

7. A method for producing a pharmaceutical composition for inhibiting the degradation of elastin fibers, characterized by containing pomegranate seed extract, comprising the steps of: immersing the resulting pulverized product in a solvent and then separating out the supernatant; obtaining a pomegranate seed extract by separating out the supernatant; and adjusting the obtained pomegranate seed extract to an effective amount.

8. 8. The method according to claim 7, wherein 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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