KR20200081914A - A composition comprising extract of Prasiola japonica for preventing or treating for inflammatory skin diseases - Google Patents

Abstract

The present invention relates to a composition for preventing or treating inflammatory skin diseases containing a Prasiola japonica extract as an active component. The Prasiola japonica extract has an excellent effect of inhibiting skin inflammation induced by ultraviolet rays, thereby being able to be usefully used as the composition for treating inflammatory skin diseases.

Description

A composition comprising extract of Prasiola japonica for preventing or treating for inflammatory skin diseases}

The present invention relates to a composition for preventing or treating inflammatory skin diseases comprising fresh water kimchi extract as an active ingredient.

Skin is a physical barrier that protects the body against a variety of environmental factors. It protects the internal organs from physical or chemical stimuli such as ultraviolet rays and plays an important role in maintaining homeostasis (Athar, M., Indian J. Exp. Biol., 40(6), 656-667, 2002), not only as a protective barrier to protect the body, but as an immune organ (Seo, WS et al., Korean Society for Biotechnology and Bioenginnering Journal, 29( 1), 36-41, 2014).

The human skin structure is largely divided into epidermis, dermis, and subcutis. The cells constituting the skin can be roughly divided into keratinocytes in the epidermal layer, melanocytes, fibroblasts in the dermal layer, and fat cells in the subcutaneous layer.

Dermal keratinocytes are components that make up most of epidermal cells and form keratin and produce various cytokines to participate in various inflammatory and immune responses (Kim, NM et al., J. Ginseng Res., 31(2), 86-92, 2007; Beissert, S. et al., Clin.Exp.Rheumatol., 24(1 Suppl 40), S1-6, 2006). When skin keratinocytes are exposed to environmental and physiological stress, an inflammatory reaction occurs, which causes TNF-α (tumor necrosis factor-α), IFN-γ (interferon-γ), IL-1β (interleukin-1β), IL It releases several types of inflammatory cytokines such as -6 (Kim, JH et al., The immunopathogenesis of psoriasis, Dermatol. Clin., 33, 13-23, 2015).

In addition, NF-κB (nuclear factor-kappa B), which plays an important role in the inflammatory response, is a transcription factor that regulates the synthesis of various cytokines, chemokines, and growth factors. NF-κB is composed of p50 and p65, enters the nucleus, acts as a transcription factor, synthesizes iNOS, COX-2 and inflammation-related cytokines, and binds NF-κB in general by binding to inhibitory NF-κBα (IκBα) in the cytoplasm. The action of is suppressed. The regulation of their activity is a key factor for controlling the inflammatory response, and studies have been actively conducted to find natural materials that modulate the activity of NF-κB to alleviate the inflammatory response (Kang Bo-kyung et al., Microbiol. Biotechnol. Lett., 43 (2), 112-119, 2015).

Ultraviolet rays, one of the sun's rays, are electromagnetic spectral rays between 200 and 400 nm. Among the ultraviolet rays, the wavelength bands directly related to the human body are UVA and UVB. When ultraviolet rays such as UVA and UVB are irradiated to the skin, vitamin D synthesis, epithelial formation, and sterilization effects are achieved by photochemical effects of proteins or nucleic acids in the body, but dermatitis, skin cancer, sunburn, pigmentation, and skin aging , Dryness, fine lines, etc.

Acute inflammatory reaction of the skin caused by UV irradiation is typical of erythema reaction caused by temporary expansion of capillaries and osmotic effects due to increased permeability of capillary walls, and the production of melanocytes increases when acute inflammatory reaction passes. As a result, pigmentation occurs, a reaction that protects the human body from harmful stimuli. If the functions of skin cells responsible for skin immunity are over-activated to secrete excessive amounts of certain immune factors, or if the function is not performed properly, psoriasis, allergic contact dermatitis (ACD), atopy Various inflammatory skin diseases, such as atopic dermatitis and vitiligo, are caused.

Steroid preparations, topical immunosuppressants, topical antibiotics, and antihistamines are currently used as treatments for inflammatory skin diseases, but when the drug is used for a long time, swelling swelling, skin atrophy, capillary expansion, steroid acne, hirsutism, purpura Side effects may occur. In particular, when steroids are applied to a wide range of areas, the drug can be absorbed systemically through the skin, which may interfere with growth in children, and in adults, systemic side effects such as osteoporosis may occur. Therefore, there is a need to develop a composition that is derived from natural materials and has fewer side effects, which is safe for the human body and has an excellent effect of improving inflammatory skin diseases.

Freshwater seaweed ( Prasiola japonica ) has been handed down under the name of water seaweed, and is known as freshwater seaweed in modern times. Unlike fresh seaweed, which is a red algae, freshwater seaweed is a green algae (Phylum Chlorophyta) that grows in freshwater, not in the sea, and is included in the freshwater paraphysis (Park, MK et al., J. Plant Bio., 13, 1-10, 1970). In Japan, freshwater seaweed is called kawanori (river stream seaweed) and artificial commercial culture of freshwater algae such as stream seaweed creates high commercial profits. Korean freshwater seaweed was first collected in 1938 by a Japanese scholar, Okada, from Muncheon-myeon, Muncheon-gun, Hamgyeongnam-do, in Prasiola formosana var. It was published in the name of coreana Okada and was published (Okada, Y., J. Jpn. Bot., 15, 449-452, 1939), and morphological and ecological studies of the Samcheok group have been conducted (Park, MK et al., J. Plant Bio., 13, 1-10, 1970). In Korea, it has been known that the species disappeared more than 40 years ago due to the artificial influence of industrialization, but it has been found that freshwater seaweed is still distributed in Socheon, Samcheok-si, Gangwon-do.

Green algae-derived extracts, such as freshwater seaweed, have been reported to have antihypertensive and UV-blocking activity by mycosporine-like amino acids (MAAs) (Cha, SH et al., J. Korean) Soc. Food Sci. Nutr., 35, 307-314, 2006; Groniger, A. et al., J. Photochem.Photobiol. B., 66, 54-59, 2002), used as freshwater-derived seaweed in Japan Suizenzinori ( Aphanothece sacrum ) has been found to be rich in minerals such as high content of iron and calcium, and has been highly evaluated for pharmacological utilization (Ngatu, NR et al., Ann.Allergy Asthma Immunol., 108 , 117-122, 2012). However, studies on the analysis of physiologically active substance components and pharmacological analysis of native freshwater seaweed have not been conducted so far.

As a prior art literature related to inflammatory skin diseases, including freshwater seaweed extract, prior literature [Seo, D. W. et al., J. Biomed. Res., 14(2), 83-90, 2013] and [Sa, J. H. et al., Rep. Inst. Health & Environ., 25, 52-62, 2014] disclosed the physiological activity (anti-cancer effect, anti-inflammatory effect, anti-oxidative effect, anti-diabetic effect, sunscreen effect) of freshwater seaweed extract, and Korean Patent Registration No. 10-1853687 The anti-inflammatory composition of the radiation pattern seaweed extract was disclosed in Korean Patent No. 10-1015702, and a composition for improving inflammation and skin irritation of seaweed extract has been disclosed. However, there have been no previous reports that the freshwater seaweed extract as in the present invention has the effect of preventing or treating inflammatory skin diseases.

Korean Registered Patent No. 10-1853687, anti-inflammatory composition comprising sterol fraction of radish seaweed extract and method of manufacturing sterol fraction of radish seaweed extract, registered on April 25, 2018. Korean Registered Patent No. 10-1015702, Composition for improving and alleviating inflammation and skin irritation, including seaweed extract, registered on February 10, 2011.

Kang Bo-Kyung et al., Anti-inflammatory effect of large leaf cap ethanol extract through inhibition of NF-κB regulation and mouse model from LPS-induced RAW 264.7 cells, Microbiol. Biotechnol. Lett., 43(2), 112-119, 2015. Athar, M., Oxidative stress and experimental carcinogenesis, Indian J. Exp. Biol., 40(6), 656-667, 2002. Beissert, S., et al., Mechanisms of immune-mediated skin diseases: an overview, Clin. Exp. Rheumatol., 24 (1 Suppl 40), S1-6, 2006. Cha, S. H. et al., Screening of extracts from marine green and brown algae in Jeju for potent marine angiotension-I converting enzyme (ACE) inhibitory activity, J. Korean Soc. Food Sci. Nutr., 35, 307-314, 2006. Groniger, A. et al., Induction of the synthesis of an UV-absorbing substance in the green alga Prasiola stipitata, J. Photochem. Photobiol. B., 66, 54-59, 2002. Kim, J. H. et al., The immunopathogenesis of psoriasis, Dermatol. Clin., 33, 13-23, 2015. Kim, N. M. et al., Effect of Korean red ginseng on collagen biosynthesis and MMP-1 activity in human dermal fibroblast, J. Ginseng Res., 31(2), 86-92, 2007. Ngatu, N. R. et al., Anti-inflammatory effects of sacran, a novel polysaccharide from Aphanothece sacrum, on 2,4,6-trinitrochlorobenzene-induced allergic dermatitis in vivo, Ann. Allergy Asthma Immunol., 108, 117-122, 2012. Okada, Y., On a new Prasiola from Corea, J. Jpn. Bot., 15, 449-452, 1939. Park, M. K. et al., Ecological and morphological studies on the Prasiola sp. in the Samchuck-Chodang, J. Plant Bio., 13, 1-10, 1970. Sa, J. H. et al., Chemical Characteristics and Physiological Activities from Freshwater Laver Grown in the Area of Samcheok-city, Rep. Inst. Health & Environ., 25, 52-62, 2014. Seo, D. W. et al., Screening of functional components derived from fresh water laver, Prasiola japonica, and its pharmacological properties, J. Biomed. Res., 14(2), 83-90, 2013. Seo, W. S. et al., Study for possibility of N,N,N-Trimethylphytosphingosine (TMP) for management of chronic skin diseases, Korean Society for Biotechnology and Bioenginnering Journal, 29(1), 36-41, 2014.

An object of the present invention is to provide a composition for preventing or treating inflammatory skin diseases comprising freshwater kimchi extract as an active ingredient.

The present invention relates to a pharmaceutical composition for the prevention or treatment of inflammatory skin diseases comprising freshwater seaweed ( Prasiola japonica ) extract as an active ingredient.

The freshwater seaweed is a green algae, and unlike red seaweed, seaweed, grows in fresh water, not in the sea, and belongs to the freshwater parasaceae.

The freshwater seaweed extract can be obtained by extracting freshwater seaweed with one or more solvents selected from the group consisting of water, lower alcohols of C1 to C4, acetone, hexane, dichloromethane and ethyl acetate, and lower alcohols of C1 to C4 The furnace may be selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol and isobutanol, preferably ethanol.

In addition, the extraction time of the extract is not particularly limited, it is preferable to extract within 10 minutes to 1 day, as an extraction device, a conventional extraction device, an ultrasonic pulverizer or a fractionator may be used.

The inflammatory skin disease is characterized by being induced by ultraviolet rays, and is specifically selected from the group consisting of psoriasis, atopic dermatitis, allergic dermatitis, seborrheic dermatitis, contact dermatitis, acne, systemic lupus erythematosus and urticaria.

The freshwater seaweed extract of the present invention may be added in an amount of preferably 0.001% to 50% by weight, more preferably 0.001% to 40% by weight, and most preferably 0.001% to 30% by weight relative to the total weight of the pharmaceutical composition. have.

The pharmaceutical composition according to the present invention may be formulated in a suitable form together with a commonly used pharmaceutically acceptable carrier. “Pharmaceutically acceptable” refers to a composition that is physiologically acceptable and does not cause an allergic reaction or similar reaction, such as gastrointestinal disorders, dizziness, etc., when administered to a human. In addition, the composition may be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injectable solutions, respectively, according to a conventional method. Preferably, it can be used in the form of oral dosage forms, sprays, gels, ointments, creams or external preparations.

Carriers, excipients and diluents that can be included in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl Cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl paraoxybenzoate, propyl paraoxybenzoate, talc, magnesium stearate, and mineral oil may be included, but is not limited thereto. In the case of formulation, it is prepared using diluents or excipients such as fillers, stabilizers, binders, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include at least one excipient in the extract of the present invention, for example, starch, microcrystalline cellulose, sucrose or lactose, It is prepared by mixing low-substituted hydroxypropyl cellulose and hypromellose. Also, in addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, intravenous solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used, various excipients, such as wetting agents, sweetening agents, fragrances, and preservatives, can be included. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations and suppositories. As non-aqueous solvents and suspensions, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin, glycerol, gelatin, etc. may be used. Adjuvants such as sterile and/or preservatives, stabilizers, hydrating or emulsifying accelerators, salts and/or buffers for osmotic pressure control, and other treatments of the extract or its pharmaceutically acceptable salts for formulation into parenteral dosage forms. It can be prepared as a solution or suspension by mixing in water with a useful substance, and it can be prepared in ampoules or vials.

The pharmaceutical composition comprising the extract disclosed in the present invention as an active ingredient may be administered to various mammals, such as rats, livestock, and humans. Any mode of administration can be envisaged, for example, oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dura or cerebrovascular injection. The dosage is the age, gender, weight of the subject to be treated, the specific disease or pathology to be treated, the severity of the disease or pathology, the time of administration, the route of administration, the absorption, distribution and excretion rate of the drug, the type of other drug used and the prescriber's It will depend on the judgment. Dosage determination based on these factors is within the level of those skilled in the art, and the dosage is generally in the range of 0.01 mg/kg/day to approximately 2000 mg/kg/day. A more preferred dosage is 1 mg/kg/day to 500 mg/kg/day. The administration may be administered once a day, or may be divided into several times. The above dosage does not limit the scope of the present invention in any way.

In another aspect, the present invention relates to a health functional food for preventing or improving inflammatory skin disease, including freshwater seaweed ( Prasiola japonica ) extract as an active ingredient.

The health functional food refers to food manufactured or processed using ingredients or ingredients having useful functionality, and includes, for example, health supplements, functional foods, nutritional supplements, and supplements.

The extract may be added in an amount of preferably 0.001% to 50% by weight, more preferably 0.001% to 30% by weight, and most preferably 0.001% to 10% by weight relative to the total weight of the dietary supplement. have. The health functional food of the present invention includes tablets, capsules, pills or liquids, and foods to which the extract of the present invention can be added, for example, various foods, beverages, gums, teas, vitamin complexes, etc. There is this.

The present invention relates to a composition for preventing or treating inflammatory skin diseases comprising fresh water kimchi extract as an active ingredient. The freshwater seaweed extract is excellent in inhibiting skin inflammation induced by ultraviolet rays, and thus can be usefully used as a composition for the treatment of inflammatory skin diseases.

1 is a result of confirming a change in mRNA expression levels of COX-2, IL-1β and IL-8 according to freshwater seaweed extract (Pj-EE) treatment in HaCaT cells in which inflammation is induced by UV light.
Figure 2 is a result confirming the mRNA expression changes of IL-6, TNF-α and IFN-γ according to freshwater seaweed extract (Pj-EE) treatment in inflammation-induced HaCaT cells by UV.
3 is a result of confirming changes in protein expression levels of IκBα, P65 and P50 and phosphorylated IκBα, P65 and P50 according to freshwater seaweed extract (Pj-EE) treatment in HaCaT cells in which inflammation is induced by UV light.

Hereinafter, a preferred embodiment of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, the contents introduced herein are thorough and complete, and are provided to sufficiently convey the spirit of the present invention to those skilled in the art.

<Example 1. Preparation of freshwater seaweed extract>

Freshwater seaweed ( Prasiola japonica ) of the present invention used freshwater seaweed collected from Samcheok City.

Freshwater seaweed collected from Samcheok City was washed with tap water to remove impurities, and sterilized using 70% [v/v] ethanol, followed by washing with distilled water and freeze-dried. The freeze-dried freshwater seaweed was finely crushed, and 1 l of 80% [v/v] ethanol was added to 100 g of freshly crushed freshwater seaweed and immersed at room temperature for 24 hours. After filtering the extract obtained by repeating this process 3 times with Whatman filter paper, the evaporator was used to evaporate the solvent to obtain a freshwater extract.

<Example 2. Culture of HaCaT cells>

Human keratinocyte cell (HaCaT) cells were cultured at 37° C. using DMEM (Dulbecco's Modified Medium) medium containing 10% fetal bovine serum (FBS), 1% penicillin, and 100 U/ml streptomycin. It was used for the experiment by subculturing in 5% CO2 condition.

<Example 3. Confirmation of skin inflammation inhibitory effect>

In order to confirm the effect of inhibiting inflammation according to the treatment of the freshwater seaweed extract in the skin cells, mRNA expression of COX-2, IL-1β, IL-6, IL-8, TNF-α and IFN-γ and IκBα, P65 And protein expression of P50.

Seeded per well such that the embodiment 8 × 10 ⁵ cells of the HaCaT cells cultured in Example 2 on a 6-well plate and cultured for 24 hours. Subsequently, 30 mJ/cm ² of UVB was irradiated with a UVB (317 nm) lamp to induce skin inflammation, and freshwater seaweed extract was treated with concentrations (50, 100, 200 µg/ml) and cultured for 24 hours. The mRNA expression of COX-2, IL-1β, IL-6, IL-8, TNF-α and IFN-γ and protein expression of IκBα, P65 and P50 were confirmed from the cells, and are shown in FIGS. 1 to 3.

Looking at Figures 1 to 3, when treated with freshwater seaweed extract in skin inflammation-induced HaCaT cells by UV treatment, COX-2, IL-1β, IL-6, IL-8, TNF-α and IFN-γ mRNA It was found that the expression and phosphorylated protein expression of IκBα, P65 and P50 are concentration-dependently reduced, thereby reducing the inflammatory response. In addition, although not shown in the figure, the freshwater seaweed extract of the present invention suppressed the inflammatory response more selectively in skin cells (HaCaT cells) than macrophages (RAW 264.7 cells). From this, it was found that the freshwater seaweed extract of the present invention can be usefully used as a composition for the treatment of inflammatory skin diseases induced by ultraviolet rays.

<Formulation Example 1. Pharmaceutical preparation>

Formulation Example 1-1. Preparation of hydrophilic ointment

Using the freshwater seaweed extract of Example 1 of the present invention, a hydrophilic ointment was prepared by a conventional method according to the composition shown in Table 1 below.

Raw material name Content (% by weight) Example 1 8.0 White vaseline 35.0 Stearyl alcohol 30.0 Ethyl (or methyl) p-oxybenzoate a very small amount Propylene glycol 20.0 Sodium lauryl sulfate 3.4 Propyl p-oxybenzoate a very small amount

Formulation Example 1-2. Preparation of tablets

Using the freshwater seaweed extract of Example 1 of the present invention, after mixing the components shown in Table 2, tablets were prepared by tableting according to a conventional tablet manufacturing method.

Raw material name Weight (mg) Example 1 100mg Corn starch 100mg Lactose 100mg Magnesium stearate 2mg

< Formulation example 2. Production of health functional food>

20 g of freshwater seaweed extract of Example 1 of the present invention, a suitable amount of vitamin mixture, 70 μg of vitamin A acetate, 1.0 mg of vitamin E, 0.13 mg of vitamin B1, 0.15 mg of vitamin B2, 0.5 mg of vitamin B6, 0.2 μg of vitamin B12, and 10 mg of vitamin C , Biotin 10 µg, Nicotinic acid amide 1.7 mg, Folic acid 50 µg, Calcium Pantothenate 0.5 mg, Mineral mixture amount, Ferrous sulfate 1.75 mg, Zinc oxide 0.82 mg, Magnesium carbonate 25.3 mg, Potassium phosphate 15 mg, Dibasic phosphoric acid 55 mg of calcium, 90 mg of potassium citrate, 100 mg of calcium carbonate, and 24.8 mg of magnesium chloride were mixed to prepare granules, but can be prepared by modifying them into various formulations depending on the application. In addition, the composition ratio of the above-mentioned vitamin and mineral mixture may be arbitrarily modified, and the above ingredients may be mixed and prepared according to a conventional method for manufacturing a health functional food.

< Formulation example 3. Production of health functional beverages>

A beverage was prepared by mixing 1 g of freshwater seaweed extract of Example 1 of the present invention, 0.1 g of citric acid, 100 g of fructooligosaccharide, and 900 g of purified water, followed by stirring, heating, filtering, sterilizing, and refrigerating according to a conventional beverage preparation method.

Claims (10)

A pharmaceutical composition for preventing or treating inflammatory skin diseases, including freshwater kim ( Prasiola japonica ) extract as an active ingredient. According to claim 1,
The freshwater seaweed extract is a pharmaceutical composition for preventing or treating inflammatory skin diseases, characterized in that freshwater seaweed is obtained by extraction and concentration with water, a lower alcohol of C1 to C4, or a mixed solvent thereof. According to claim 1,
The inflammatory skin disease is a pharmaceutical composition for preventing or treating inflammatory skin disease, characterized in that induced by ultraviolet rays. According to claim 1,
The inflammatory skin disease is psoriasis, atopic dermatitis, allergic dermatitis, seborrheic dermatitis, contact dermatitis, acne, systemic erythematous lupus and urticaria inflammatory skin disease prevention or treatment pharmaceutical composition characterized in that selected from the group consisting of. According to claim 1,
The pharmaceutical composition of the pharmaceutical composition for the prevention or treatment of inflammatory skin diseases, characterized in that the formulation is selected from the group consisting of oral dosage forms, sprays, gels, ointments, creams or external preparations. A health functional food for preventing or improving inflammatory skin diseases, including freshwater kim ( Prasiola japonica ) extract as an active ingredient. The method of claim 6,
The freshwater seaweed extract is a health functional food for preventing or improving inflammatory skin diseases, characterized in that freshwater seaweed is obtained by extracting and concentrating with water, a lower alcohol of C1 to C4, or a mixed solvent thereof. The method of claim 6,
The inflammatory skin disease is a health functional food for preventing or improving inflammatory skin disease, characterized in that induced by ultraviolet rays. The method of claim 6,
The inflammatory skin disease is psoriasis, atopic dermatitis, allergic dermatitis, seborrheic dermatitis, contact dermatitis, acne, systemic lupus erythematosus lupus and urticaria, characterized in that selected from the group consisting of inflammatory skin disease prevention or improvement health functional food . The method of claim 6,
The formulation of the health functional food is a health functional food for preventing or improving inflammatory skin disease, characterized in that selected from the group consisting of tablets, capsules, pills or liquids.

Images