KR101694662B1 - Manufacturing methods of ultrasonicating extract of Star Reindeer Lichen with anti-inflammatory effect - Google Patents

Abstract

The present invention relates to a method for preparing an extract of Astragalus licheniformis having an anti-inflammatory effect, comprising the steps of: preparing a Astragalus licheniformis extract; The method of the present invention is characterized by the step of confirming the antioxidant activity, the anti-inflammatory activity and the wrinkle-reducing activity of the extract, and the starch moss ultrasound extract prepared according to the present invention has excellent antioxidative and anti-inflammatory effects and synergistically improved wrinkles.

Description

{Manufacturing methods of ultrasonicating extract of Starch lichen with anti-inflammatory effect}

The present invention relates to a method for producing an ultrasound extract of Star Reindeer Lichen having an anti-inflammatory effect.

Moss is a small, soft plant belonging to the lichen or tideland, usually 1 to 10 cm, but much larger. Mosses are usually grouped together in moist, shady places, do not have flowers or seeds, but sometimes produce sporangia. In addition, moss is known to have a strong vitality to survive in outer space (Goffinet, B. et al., Morphology and Classification of the Bryophyta, ryophyte Biology, Cambridge University Press, 55-138, 2008). Moss is also called lichen, and Cladonia stellaris among lichens is called Star Reindeer Lichen moss and is called Korean mackerel (mangosteen moss). There are about 400 species of Cladonia genus in the world and 30 species in the King George Island of Antarctica, but the research is insufficient (Lee, JS et al., Diversity of the Lichenized Fungi in King George Island, Antarctica, Revealed by Phylogenetic Analysis of Partial Large Subunit rDNA Sequences. Journal of Microbiology and Biotechnology, 18, 1016-1023, 2008).

Skin aging is divided into intrinsic aging and extrinsic aging. Inner aging is the decline of the structure and physiological function of the skin with age, while external aging is caused by external aging such as sunlight Gilchrest BA Skin aging and photoaging, Dermatol Nurs 2, 79-82, 1990). Skin aging caused by ultraviolet rays is caused by skin cells producing melanin pigment for the purpose of defending against ultraviolet absorption (Wang KH et al., Cosmetic applications of selected traditional Chinese herbal medicines, J. Ethnopharmacol, 353-359, 2006). Many studies have shown that these skin troubles are also associated with active oxygen. Active oxygen destroys collagen, elastin, and hyaluronic acid, which are connective tissues of the dermis, causing wrinkles of the skin, oxidizing the lipid part of the cell membrane, causing cell destruction, and causing diseases such as dermatitis, acne and skin cancer. In addition, this active oxygen participates in the spontaneous oxidation reaction during the melanin formation process, and causes the causes such as stain and freckles and wrinkle formation (Korean Journal of Cosmetic Science, Vol. 23, No. 1, 75-132). As skin aging progresses, the ability to produce structural proteins such as collagen, elastin, and hyaluronic acid, which constitute the skin, decreases and collagenase biosynthesis increases, resulting in increased expression of matrix metalloproteinases (MMPs) , fibronectin and laminin, resulting in skin elasticity and skin wrinkle formation (Brenneisen P, et al., Ultraviolet-B irradiation and matrix metalloproteinases: from induction via signaling to initial events, Ann. NY Acad. Sci. 31-43, 2002). Vitamin C, arbutin, ascorbic acid, kojic acid, hydroquinone, and retinol collagen are used as various materials effective for improving skin aging. However, there are constant problems in product safety.

A cosmetic composition comprising a moss extract, wherein the composition of Cladonia An anti-aging dermatological composition comprising an extract of n-hexane of stellaris or an active ingredient thereof, depsidone or a depsidone derivative is disclosed in Korean Patent Laid-Open No. 10-2009-0052096, Korean Patent No. 10-1176521 discloses a cosmetic composition for improving skin comprising, as an active ingredient, a mixed extract of moss, octagonal fennel, echinacea and propolis. In addition, Korean Patent No. 10-1083347 discloses a pharmaceutical composition for improving or treating atopic dermatitis using moss extract. However, no method has been disclosed in the above-mentioned literature for producing a starchy moss extract having an antiinflammatory effect by adding a polar solvent to Star Reindeer Lichen among lichens (lichen) and ultrasonically extracting it.

Accordingly, an object of the present invention is to provide a method for producing an ultrasound extract of Star Reindeer Lichen which has anti-inflammatory effect.

The above object of the present invention can be achieved by a method for producing an extract of the present invention comprising the steps of: Antioxidant activity, antiinflammatory activity and collagen aggregation performance of the extract were compared and confirmed.

In the present invention, when the extract of Star Reindeer Lichen is combined with the water and the organic solvent mixture by the ultrasonic extraction method, the antioxidative activity and the anti-inflammatory activity are increased and the wrinkle is improved.

FIG. 1 is a graph showing the antioxidative effect of Star Reindeer Lichen extract of the present invention with time. FIG.
FIG. 2 is a graph showing the antioxidative effect of 0.5%, 1.0% and 2.0% of the extract of Star Reindeer Lichen extract of the present invention.
FIG. 3 is a graph showing the antioxidative effect of 0.5%, 1.0% and 2.0% of the solvent extract of Star Reindeer Lichen according to the present invention.
FIG. 4 is a graph showing the anti-inflammatory effect of 0.5%, 1.0%, and 2.0% of the moss ectopic extract of the present invention.
FIG. 5 is a graph showing the collagen aggregation performance of 0.5%, 1.0%, and 2.0% of the moss ectopic extract of the present invention.

The present invention provides a method for producing an ultrasound extract of Star Reindeer Lichen having anti-inflammatory activity.

The extract of the present invention can be obtained only by extraction with water, an organic solvent or a mixed solvent thereof. The organic solvent used herein is ethanol, butylene glycol, propylene glycol, propanediol, and the like. It may also be a dried product obtained by drying the extract obtained by the above extract, a diluted solution of the extract, a concentrate or an extract, or a preparation or a purified product of the extract. However, in the present invention, it is most preferable to extract propolis diol by adding propanediol to a shrub specimen and extracting it using an ultrasonicator, wherein the extract is fixed at a temperature of 40 ° C and then subjected to ultrasonic treatment.

In the present invention, the above-prepared astringent moss ultrasound extract is characterized by containing 0.1 to 5% by weight, preferably 2% by weight, of the total weight of the cosmetic composition. When the content is less than 0.1% by weight, the active oxygen species inhibiting effect of the present invention is insufficient. When the content of the active ingredient is more than 5% by weight, inherent odor and color depending on the unique flavor are strong.

According to the present invention, it is necessary that rupture of the cell membrane by ultrasonic treatment is absolutely necessary because the embryonic cell tissue is hard. In order to improve the wrinkles caused by aged skin damage as in the present invention, it is necessary to select a polar solvent without extracting the active ingredient from the cell material but without removing the solvent.

Propanediol, which is capable of maximizing the extraction efficiency of the cell material without polar solvent removal, is preferably selected.

According to the present invention, it is most preferable to maximize the extraction of the flavonoid substance or the polyphenol component when the organic solvent is added and to treat the ultrasonic wave, and furthermore, the temperature is 35 to 40 ° C at which there is no thermal denaturation of the protein enzyme.

The cosmetic composition having a skin protecting effect according to the present invention may have conventional formulations and may be in the form of, for example, water-soluble liquid, lotion, cream, gel, pack, essence, oil / water, ) Or a silicone heavy (W / Si) type powder. The composition of each of these formulations may contain various kinds of bases and additives necessary for formulation of the formulation, and the kinds and amounts of these ingredients can be easily selected by those skilled in the art.

In addition, the cosmetic composition containing the extract of Aspergillus licheniformis having antioxidative, antiinflammatory and wrinkle-reducing effects according to the present invention can be used according to the conventional method of use, and the number of times of use thereof Can be different.

Hereinafter, the present invention will be described in more detail by way of examples. However, the following examples are intended to illustrate the invention and are not intended to limit the scope of the invention.

< Example > Invention Moss flower Propanediol  Manufacture of Ultrasonic Extracts

40 ~ 60 mL of 40% propanediol was added to 2 ~ 3g of shiitake moss, and the temperature was fixed at 35 ~ 40 ℃ or less using an ultrasolificator (JAC 4020, Jinwoo, Korea) We obtained ultrastructural extract of mackerel moss. All the procedures were repeated three times for 1 hour. The extracted samples were filtered with a 0.45 μm membrane filter, and then subjected to an efficacy test.

< Comparative Example  > Moss flower Propanediol  Preparation of extract

40 ~ 60 mL of 40% propanediol was added to 2 ~ 3 g of dried starch moss and extracted at 40 ° C to obtain an organic solvent extract. All the procedures were repeated three times for 1 hour. The extracted samples were filtered with a 0.45 μm membrane filter, and then subjected to an efficacy test.

< Experimental Example  1> Moss flower  Antioxidant activity test of extract

In this experimental example, the skin fibroblast (HDFn) was suspended in DMEM (containing 10% serum and 1% antibiotic), cultured in a 5% CO ₂ incubator at 37 ° C, .

Wells were inoculated in 96 wells at a concentration of about 1 x 10 ⁵ cells per well. The starch moss supernatant extract prepared according to the above Example and the starch moss hot water extract prepared according to the Comparative Example were suspended in 0.5%, 1% and 2% And cultured in a 5% CO ₂ incubator at 37 ° C for 20 minutes. 1 mM H ₂ O ₂ was added and 20 μM DCFH-DA (stock 20 mM) was added. The reaction was monitored by a spectrofluorophotometer (excitation 485 nm, emission 535 nm) at 37 ° C. for 90 minutes . The inhibition rate of reactive oxygen species was determined by comparing the formation of reactive oxygen species with the absence of the sample as a control group, and compared with the group treated with a sample to remove active oxygen species. All experiments were repeated 3 times.

As a result of the experiment, as shown in Fig. 1, The antioxidant activity of the extracts increased with time. As shown in FIG. 3, when organic solvent was added to the moss moss, the extract was inhibited by 29% at a maximum concentration of 2%, and 19% at a maximum concentration of 2% Respectively. Therefore, the extraction efficiency of organic solvent was 10% higher than that of organic solvent extraction.

< Experimental Example  2> invention Moss flower  Nitric Oxide of Ultrasonic Extract Low performance  Experiment

In this experiment, LPS (Lipopolysaccharide), an inflammation inducing substance, was artificially treated with mouse macrophage cell RAW 264.7 cell, and then it was evaluated whether or not it has an inflammation inhibitory effect.

Cells were seeded at 2 × 10 ⁵ cells per well in a 96-well plate and cultured in a 5% CO ₂ incubator at 37 ° C for 24 hours. LPS (1 ug / mL) and extracts were added at different concentrations and incubated for 24 hours. After incubation, the supernatant was taken and centrifuged at 13,000 rpm for 3 minutes. The supernatant was collected, reacted with Griess reagent 1: 1 and absorbance was measured at 570 nm.

As a result, as shown in FIG. 4, the Nitric Oxide inhibitory activity of the present invention was higher than that of the control group.

< Experimental Example  3> invention Moss flower  Collagen of ultrasound extract Sum performance  Experiment

In this experiment, HDFn was inoculated into a culture medium (0.3 mL) at a concentration of 5 × 10 ⁴ cells / well in a 48-well plate and cultured at 37 ° C and 5% CO ₂ for 24 hours. And further cultured for 24 hours in a fresh medium containing the flower moss ultrasound extract of the present invention at different concentrations. The cultures were collected and the amount of C-terminus of the collagen precursor was measured using an enzymes-linked immunoassay kit (Takara). A standard curve between collagen concentration of 0 ng and 640 ng was obtained using collagen solution (640 ng / The amount of collagen in the medium was calculated.

As a result, as shown in FIG. 5, the collagen synthesis performance was increased with increasing concentration of the present invention. Especially, the collagen synthesis performance was increased by 128% when 2% of the ultrasound extract was treated.

INDUSTRIAL APPLICABILITY As described above, the present invention is an extremely useful invention in the cosmetics industry because it has an effect of providing an ultrasound extract derived from a mosquito net with anti-inflammatory effect.

Claims (1)

Characterized in that 40 to 60 mL of propanediol is added to 2 to 3 g of a shaded venomous moss, and the mixture is sonicated at a temperature of 35 to 40 DEG C for 1 hour.

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