KR102261340B1 - 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 comprising a freshwater laver extract as an active ingredient. Since the freshwater laver extract is excellent in inhibiting skin inflammation induced by ultraviolet rays, it can be usefully used as a composition for treating inflammatory skin diseases.

Description

A composition comprising extract of Prasiola japonica for preventing or treating for inflammatory skin diseases, comprising extract of freshwater laver as an active ingredient

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

As a physical barrier that protects the body against various environmental factors, the skin protects 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), it is considered not only as a protective film to protect the body, but also as an immune organ (Seo, WS et al., Korean Society for Biotechnology and Bioenginnering Journal, 29 (Seo, WS et al., Korean Society for Biotechnology and Bioenginnering Journal, 29) 1), 36-41, 2014).

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

Skin keratinocytes are a component that constitutes most of the epidermal cells, and are involved in various inflammatory and immune responses by forming keratin and producing various cytokines (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 response occurs, resulting in TNF-α (tumor necrosis factor-α), IFN-γ (interferon-γ), IL-1β (interleukin-1β), IL It secretes 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 and acts as a transcription factor, synthesizes iNOS, COX-2, and inflammation-related cytokines, and generally binds to IκBα (inhibitor NF-κBα) in the cytoplasm to bind NF-κB action is inhibited. Their activity regulation is a key factor for regulating the inflammatory response, and studies are being actively conducted to find natural materials that relieve inflammatory responses by regulating the activity of NF-κB (Bokyung Kang, et al., Microbiol. Biotechnol. Lett., 43 (2), 112-119, 2015).

Ultraviolet rays, one of the sun's rays, are rays in the form of electromagnetic spectrum between 200 and 400 nm. Among 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 effect are achieved by photochemical action of proteins or nucleic acids in the body, but dermatitis, skin cancer, sunburn, pigmentation, and skin aging , dryness, and fine wrinkles may also occur.

The acute inflammatory reaction of the skin caused by UV irradiation is representative of the erythema reaction caused by the temporary expansion of capillaries and the osmosis by increasing the permeability of the capillary wall. When the acute inflammatory reaction passes, the production of melanocytes increases. Pigmentation, a reaction that protects the human body from harmful stimuli, occurs. When the function of skin cells responsible for skin immunity is overactivated and excessive secretion of specific immune factors or fails to perform their functions properly, psoriasis, allergic contact dermatitis (ACD), atopic dermatitis Various inflammatory skin diseases such as atopic dermatitis and vitiligo are caused.

Currently, steroid preparations, topical immunosuppressants, topical antibiotics, and antihistamines are used as therapeutic agents for inflammatory skin diseases. However, long-term use of the above drug preparations causes fattening striae, skin atrophy, capillary dilatation, steroid acne, hirsutism, and purpura. side effects may occur. In particular, when steroids are applied to a wide area, the drug is systemically absorbed through the skin, which can impede growth in children and systemic side effects such as osteoporosis in adults. Therefore, there is a need for the development of a composition derived from a natural substance, which has few side effects, is safe for the human body and has an excellent effect of improving inflammatory skin disease.

Freshwater laver ( Prasiola japonica ) has been handed down under the name of water laver, and is now known as freshwater laver. Unlike sea laver, which is a red alga, freshwater laver is a green algae (Phylum Chlorophyta) that grows in fresh water rather than the sea, and is included in the Prasiolacease family (Park, MK et al., J. Plant Bio., 13, 1-10, 1970). In Japan, freshwater laver is called kawanori (stream laver), and freshwater algae such as river laver are artificially cultured to create high commercial profits. Korean freshwater laver was first collected in 1938 by a Japanese scholar Okada from Muncheon-myeon, Muncheon-gun, Hamgyeongnam-do, and Prasiola formosana var. It was named coreana Okada and announced at the beginning (Okada, Y., J. Jpn. Bot., 15, 449-452, 1939), and morphological and ecological studies on the Samcheok-gun group have been conducted (Park, MK et al., J. Plant Bio., 13, 1-10, 1970). In Korea, it is known that the species disappeared 40 years ago due to the artificial influence of industrialization, but it was found that freshwater laver is still distributed in Sohancheon in Samcheok, Gangwon-do.

Extracts derived from green algae such as freshwater laver have been reported to have antihypertensive and UV protection by mycosporine-like amino acids (MAAs), etc. (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 edible laver derived from freshwater in Japan Suizenzinori ( Aphanothece sacrum ) has been investigated for being rich in minerals such as high iron and calcium, and its pharmacological utility has been highly evaluated (Ngatu, NR et al., Ann. Allergy Asthma Immunol., 108). , 117-122, 2012). However, studies on the physiologically active substance analysis and pharmacological analysis of domestic freshwater laver have not been conducted yet.

As prior prior art related to inflammatory skin disease including freshwater laver 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 activities (anticancer effect, anti-inflammatory effect, antioxidant effect, antidiabetic effect, UV protection effect) of freshwater laver extract, and Korean Patent No. 10-1853687 No. discloses an anti-inflammatory composition of a radish laver extract, and Korean Patent No. 10-1015702 discloses a composition for improving inflammation and skin irritation of a seaweed extract. However, there is no previous report mentioning that the freshwater laver extract is effective in preventing or treating inflammatory skin diseases as in the present invention.

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

Bo-Kyung Kang, et al., Inhibition of NF-κB regulation from LPS-induced RAW 264.7 cells and anti-inflammatory effect of ethanol extract of Capricornus larvae in mouse model, 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.

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

The present invention relates to a pharmaceutical composition for preventing or treating inflammatory skin diseases, comprising an extract of freshwater laver ( Prasiola japonica ) as an active ingredient.

The freshwater laver is a green algae and, unlike sea laver, which is a red algae, grows in freshwater rather than the sea, and belongs to the family Prasiolacease.

The freshwater laver extract can be obtained by extracting freshwater laver with at least one solvent selected from the group consisting of water, C1 to C4 lower alcohol, acetone, hexane, dichloromethane and ethyl acetate, and the C1 to C4 lower alcohol The furnace may be selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol and isobutanol, and ethanol is preferably used.

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

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

The freshwater laver 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 based on 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 normally cause allergic reactions such as gastrointestinal disorders, dizziness, or similar reactions when administered to humans. In addition, the composition may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, and sterile injection solutions according to conventional methods, respectively. Preferably, it can be used in formulations for oral administration, spray, gel, ointment, cream or external use.

Carriers, excipients and diluents that may 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, but is not limited thereto. In the case of formulation, it is prepared using diluents or excipients, such as commonly used fillers, stabilizers, binders, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and 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, hypromellose, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid formulations for oral use include suspensions, internal solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerol, gelatin, etc. may be used. In order to formulate a formulation for parenteral administration, the extract or a pharmaceutically acceptable salt thereof is sterilized and/or an adjuvant such as a preservative, a stabilizer, a wetting agent or emulsification accelerator, a salt and/or a buffer for regulating osmotic pressure, and other treatments It can be prepared as a solution or suspension by mixing it with water with useful substances, and it can be prepared in ampoules or vial unit dosage form.

The pharmaceutical composition comprising the extract disclosed in the present invention as an active ingredient may be administered to mammals such as mice, livestock, and humans by various routes. Any mode of administration can be envisaged, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebrovascular injection. The dosage may vary depending on the age, sex, weight, specific disease or pathology to be treated, the severity of the disease or pathology, administration time, administration route, absorption, distribution and excretion rate of the drug, the type of other drugs used, and the prescriber's It will depend on judgment, etc. Dosage determination based on these factors is within the level of the skilled artisan, and dosages generally range from 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. Administration may be administered once a day, or may be administered in several divided doses. The above dosage does not limit the scope of the present invention in any way.

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

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

The extract is preferably 0.001% to 50% by weight, more preferably 0.001% to 30% by weight, most preferably 0.001% to 10% by weight based on the total weight of the health functional food. have. The health functional food of the present invention includes the form of tablets, capsules, pills or liquids, and the like, and the food to which the extract of the present invention can be added includes, for example, various foods, beverages, gum, tea, vitamin complex, etc. There is this.

The present invention relates to a composition for preventing or treating inflammatory skin diseases comprising a freshwater laver extract as an active ingredient. Since the freshwater laver extract is excellent in inhibiting skin inflammation induced by ultraviolet rays, it can be usefully used as a composition for treating inflammatory skin diseases.

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

Hereinafter, preferred embodiments 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, it is provided so that this disclosure will be thorough and complete, and will fully convey the spirit of the invention to those skilled in the art.

<Example 1. Preparation of freshwater laver extract>

Freshwater laver ( Prasiola japonica ) of the present invention was used freshwater laver collected in Samcheok-si.

Freshwater laver collected in Samcheok was washed with tap water to remove impurities, sterilized using 70% [v/v] ethanol, washed with distilled water, and freeze-dried. Finely crushed freeze-dried laver, 1 liter of 80% [v/v] ethanol was added to 100 g of crushed laver, and immersed at room temperature for 24 hours. After repeating this process three times, the obtained extract was filtered with Whatman filter paper, and then the solvent was evaporated using an evaporator to obtain a freshwater laver extract.

<Example 2. Culture of HaCaT cells>

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

<Example 3. Confirmation of skin inflammation inhibitory effect>

In order to confirm the anti-inflammatory effect of the present invention freshwater laver extract treatment in skin cells, the mRNA expression of COX-2, IL-1β, IL-6, IL-8, TNF-α and IFN-γ and IκBα, P65 and P50 protein expression.

The HaCaT cells cultured in Example 2 were aliquoted in a 6-well plate so that 8×10 ⁵ cells per well were cultured for 24 hours. ^{Thereafter, skin inflammation was induced by irradiating UVB of 30mJ/cm 2} with a UVB (317nm) lamp, and the freshwater laver extract was treated with different 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 .

Referring to FIGS. 1 to 3, when the HaCaT cells induced by UV treatment were treated with the freshwater laver extract, mRNA of COX-2, IL-1β, IL-6, IL-8, TNF-α and IFN-γ It was found that the inflammatory response was reduced by the concentration-dependent reduction of the expression and phosphorylated protein expression of IκBα, P65 and P50. In addition, although not shown in the drawings, the freshwater laver extract of the present invention more selectively inhibited the inflammatory response in skin cells (HaCaT cells) than macrophages (RAW 264.7 cells). From this, it was found that the freshwater laver 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 formulation>

Formulation Example 1-1. Preparation of hydrophilic ointment

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

Raw material name content (wt%) 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. manufacture of tablets

Using the freshwater laver extract of Example 1 of the present invention, the ingredients shown in Table 2 were mixed, and then 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. Manufacturing of health functional food>

20 g of freshwater laver extract of Example 1 of the present invention, appropriate 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, 10 mg of vitamin C , biotin 10㎍, nicotinic acid amide 1.7mg, folic acid 50㎍, calcium pantothenate 0.5mg, mineral mixture appropriate amount, ferrous sulfate 1.75mg, zinc oxide 0.82mg, magnesium carbonate 25.3mg, potassium monophosphate 15mg, diphosphate dibasic Calcium 55 mg, potassium citrate 90 mg, calcium carbonate 100 mg, and magnesium chloride 24.8 mg were mixed to prepare granules, but it can be prepared by transforming into various formulations depending on the use. In addition, the composition ratio of the vitamin and mineral mixture may be arbitrarily modified, and it may be prepared by mixing the above ingredients according to a conventional health functional food manufacturing method.

< Formulation example 3. Manufacture of health functional beverage>

1 g of freshwater laver extract of Example 1 of the present invention, 0.1 g of citric acid, 100 g of fructooligosaccharide, and 900 g of purified water were mixed and stirred, heated, filtered, sterilized, and refrigerated according to a conventional beverage preparation method to prepare a beverage.

Claims (10)

In the pharmaceutical composition for preventing or treating inflammatory skin disease comprising freshwater laver (Prasiola japonica) extract as an active ingredient,
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 freshwater laver extract is a pharmaceutical composition for preventing or treating inflammatory skin diseases, characterized in that it is obtained by extracting and concentrating freshwater laver with water, C1 to C4 lower alcohol, or a mixed solvent thereof. delete delete According to claim 1,
The pharmaceutical composition for the prevention or treatment of inflammatory skin disease, characterized in that the formulation of the pharmaceutical composition is selected from the group consisting of a spray, a gel, an ointment, a cream, or an external application. delete delete delete delete delete

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