KR102145297B1 - Antiinflammatory and antibaterial compositions comprising extract of artemisiae annuae herb, ginkgo biloba and ficuscarica l. - Google Patents

Abstract

An embodiment of the present invention provides an antibacterial and anti-inflammatory composition comprising a blueberry extract, a ginkgo leaf extract, and a fig extract as active ingredients. Specifically, the composition according to the present invention may have an antibacterial effect against acne bacteria, and may also have an anti-inflammatory effect of suppressing inflammation. According to another embodiment of the present invention, the composition according to the present invention may reduce skin Demodex or inhibit survival.

Description

An antibacterial and anti-inflammatory composition containing blueberry extract, ginkgo leaf extract, and fig extract as active ingredients {ANTIINFLAMMATORY AND ANTIBATERIAL COMPOSITIONS COMPRISING EXTRACT OF ARTEMISIAE ANNUAE HERB, GINKGO BILOBA AND FICUSCARICA L.}

The present invention relates to an antibacterial and anti-inflammatory composition containing a natural plant extract as an active ingredient, and more particularly, to an antibacterial and anti-inflammatory composition comprising a green lake extract, a ginkgo leaf extract and a fig extract as an active ingredient.

Although acne is a common disease, the cause of its occurrence has not yet been determined.After puberty, the cause of the occurrence of acne has not been determined. Increasing the secretion of sebum due to hormonal effects after puberty, abnormally increased infundibular hyperkeratosis, and Propionibacterium acnes (P. ), etc. are believed to be the main cause. The number of bacteria P.acnes, which is considered to be the main cause of inflammatory acne, was found to have a greater number of acne lesions in adolescent acne patients than non-lesion areas, and the number of bacteria and the degree of clinical deterioration appeared to be correlated. Therefore, antibiotics that are currently used as major treatments for acne are treatments that focus on P.acnes bacteria. The mechanism of what action of P. acnes causes acne is still unclear, but according to the results of studies so far, it is expected that P. acnes is involved in both the innate and acquired immune systems and causes inflammation. It is known that P.acnes acts on histiocytes and neutrophils, which are important cells for innate immunity, to generate inflammatory cytokines, and toll-like receptor 2 (TLR2) acts in this process. In other words, TLR2, the main mediator of innate immunity caused by P.acnes, is involved in the pathogenesis of acne.

To treat acne formed by acne bacteria, antibiotics such as tetracycline, erythromycin, clindamycin, macrolides, penicillin, and cephalosporin have been used for decades. As a result, acne bacteria resistant to erythromycin and clindamycin were discovered by the Crawford research team in 1979, followed by acne bacteria resistant to tetracycline in the early 1980s. As a result of continuing research on antibiotic-resistant acne-resistant bacteria around the world, in the late 1970s, the proportion of acne bacteria resistant to erythromycin, clindamycin and tetracycline antibiotics was about 20%, but in 1995, the emergence of resistant bacteria was up to 72.5%. Furthermore, mixed resistant bacteria to erythromycin and clindamycin were more frequent than tetracycline-resistant bacteria. In the case of acne bacteria, resistance to antibiotics can be observed even if antibiotics are used for only 8 weeks, and it can be confirmed that the number of resistant bacteria gradually increases within a few weeks. As such, the emergence and spread of antimicrobial resistant bacteria is serious, and multidrug resistant bacteria that are resistant to almost all types of antimicrobial treatments developed and used to date have appeared, and the development of new compositions to replace antibiotics is urgently and urgently required. have.

The technical problem to be achieved by the present invention is to provide an anti-bacterial and anti-inflammatory composition comprising Cheongho extract, ginkgo biloba extract, and fig extract as active ingredients, as well as providing an anti-inflammatory and antibacterial composition that is safe for use in the human body.

The technical problem to be achieved by the present invention is not limited to the technical problems mentioned above, and other technical problems that are not mentioned can be clearly understood by those of ordinary skill in the technical field to which the present invention belongs from the following description. There will be.

In order to achieve the above technical problem, an embodiment of the present invention provides an antibacterial and anti-inflammatory composition comprising blueberry extract, ginkgo leaf extract, and fig extract as active ingredients.

In an embodiment of the present invention, the composition may include 20 to 40 parts by weight of the green lake extract, 20 to 40 parts by weight of the ginkgo leaf extract, and 10 to 20 parts by weight of the fig extract.

In an embodiment of the present invention, the extract is water, anhydrous or aqueous lower alcohol having 1 to 4 carbon atoms, acetone, petroleum ether, ethyl acetate, butyl acetate, trichloromethane, dichloromethane, chloroform, hexane, and 1,3- It may be extracted with one or more solvents selected from the group consisting of butylene glycol and propanediol.

In an embodiment of the present invention, the extract may be extracted by ultrasonic extraction under the solvent.

In an embodiment of the present invention, the composition may have an inhibitory function against Propionibacterium acnes.

In an embodiment of the present invention, the composition may have a function of inhibiting NF-kB activity.

In an embodiment of the present invention, the composition may be a cosmetic composition.

According to an embodiment of the present invention, a composition comprising a blueberry extract, a ginkgo leaf extract, and a fig extract as active ingredients may have antibacterial and anti-inflammatory effects. Specifically, the composition according to an embodiment of the present invention may have an antibacterial effect against acne bacteria.

The effects of the present invention are not limited to the above effects, and should be understood to include all effects that can be deduced from the configuration of the invention described in the detailed description or claims of the present invention.

Figure 1 shows the antibacterial effect of the composition according to an embodiment of the present invention against acne bacteria.
2 is a graph showing the growth inhibitory ability against acne bacteria of the composition according to an embodiment of the present invention.
Figure 3 shows the change in the shape of acne bacteria according to the composition treatment according to an embodiment of the present invention.
Figure 4 is a graph showing the ability to inhibit NF-kb activity according to the composition treatment according to an embodiment of the present invention.

Hereinafter, the present invention will be described with reference to the accompanying drawings. However, the present invention may be implemented in a number of different forms, and therefore is not limited to the embodiments described herein. In the drawings, parts irrelevant to the description are omitted in order to clearly describe the present invention, and similar reference numerals are attached to similar parts throughout the specification.

Throughout the specification, when a part is said to be "connected (connected, contacted, coupled)" with another part, it is not only "directly connected", but also "indirectly connected" with another member in the middle. "Including the case. In addition, when a part "includes" a certain component, it means that other components may be further provided, not excluding other components unless otherwise specified.

The terms used in the present specification are only used to describe specific embodiments, and are not intended to limit the present invention. Singular expressions include plural expressions unless the context clearly indicates otherwise. In the present specification, terms such as "comprise" or "have" are intended to designate the presence of features, numbers, steps, actions, components, parts, or combinations thereof described in the specification, but one or more other features. It is to be understood that the presence or addition of elements or numbers, steps, actions, components, parts, or combinations thereof, does not preclude in advance.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings.

According to one aspect of the present invention, the present invention can provide an antibacterial and anti-inflammatory composition comprising a blueberry extract, a ginkgo leaf extract, and a fig extract as active ingredients.

The “cheongho (Artemisiae Annuae Herba)” refers to the above-ground part of Artemisiaannua Linne or Artemisia apiacea Hance of the dicotyledonous plant Campanula. Dogdung wormwood is also called Swallow Mugwort, Fine Leaf Mugwort, Chogo, Hyanggo, Dog Tangerine Mugwort, Mugwort, etc. In oriental medicine, it is used for the treatment of fever, cold, dysentery, dyspepsia, dyspepsia. In addition, the wormwood wormwood is known as a source of artemisinin, an antimalarial agent.

The leaves of the "Ginkgo biloba L" contain various ingredients. The ginkgo leaves contain flavonoids, biflavonoids, proanthocyanidins, terpenlactones, etc. as major components, and bioflavonoids such as quercetin, camperol, and isohamnetin become fallen leaves. It is known to contain abundantly the most mature green leaves when harvested from September to October. The ginkgo biloba extract is known to inhibit the expression of melasma-like blood vessels, which are found in a lot of toxic spots, to prevent abnormal melanocytes from being activated.

The “ficus (Ficuscarica L.)” is a subtropical semi-arboreal deciduous broad-leaved tree and belongs to the Morus family. More than 600 varieties are distributed worldwide, and it is known as the first fruit used by humans in a long history. The fig has been recorded in the Bible or Donguibogam for civilian medical use since ancient times, and is known to be good for strengthening blood pressure, health, nutrition, constipation, hepatitis, cancer, and recovery of vitality from women's diseases. The general effects of figs include gastrointestinal action, treatment of diarrhea or hemorrhoids, sore throat, anti-inflammatory action, minor action, digestion promotion, etc., and are also used for anthelmintic medicine and jaundice.

The "extract" refers to a solvent in which the active ingredient contained in the raw material for extraction is transferred by contacting the solvent with the raw material for extraction under specific conditions. You can include all regardless. For example, it may include extracting a component soluble in a solvent from a natural product using water or an organic solvent, and extracting a specific component of a natural product, for example, a specific component such as oil. The extract may be obtained by drying and powdering the extraction raw material, or by various conventionally known extraction methods such as hot water extraction and ethanol extraction.

According to an embodiment of the present invention, the composition may include 20 to 40 parts by weight of the blueberry extract, 20 to 40 parts by weight of the gingko leaf extract, and 10 to 20 parts by weight of the fig extract. Preferably, it may include 20 to 30 parts by weight of the blueberry extract, 20 to 30 parts by weight of the gingko leaf extract, and 10 to 15 parts by weight of the fig extract. More preferably, it may include 20 to 25 parts by weight of the blueberry extract, 20 to 25 parts by weight of the gingko leaf extract, and 10 to 12 parts by weight of the fig extract.

According to an embodiment of the present invention, the extract is water, an anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms, acetone, petroleum ether, ethyl acetate, butyl acetate, trichloromethane, dichloromethane, chloroform, hexane and 1,3 -It can be extracted with one or more solvents selected from the group consisting of butylene glycol and propanediol. Specifically, it may be extracted with 60 to 1,3-butylene glycol and propanediol.

The extract is extracted by washing the raw material with water, drying and pulverizing, and using a solvent having a volume of 8 to 12 times the weight of the raw material for about 1 hour to 24 hours by a conventional extraction method such as reflux extraction, pressure extraction, ultrasonic extraction, etc. It can be prepared by extraction and filtration. According to an embodiment of the present invention, preferably, the extract may be extracted by an ultrasonic extraction method. In addition, the extract can be obtained in a powder state by an additional process such as distillation under reduced pressure or freeze drying.

According to another embodiment of the present invention, the composition may have an inhibitory function against Propionibacterium acnes. The acne bacteria may cause skin troubles, and such skin troubles may be inflammation, acne, comedones, papules, rashes, pus blisters, nodules, cysts, and false sacs.

According to an embodiment of the present invention, the composition may have a function of inhibiting NF-kB activity. The NF-kB is a protein complex capable of regulating DNA transcription, cytokine production and cell survival, and is found in almost all cells. Referring to Figure 4, as a result of confirming the composition according to an embodiment of the present invention through the Luciferase assay, it was confirmed that it can inhibit the activity of NF-kB.

According to another embodiment of the present invention, the composition may further include a skin demodex removal function. The cutaneous demodex is an ectoparasite observed under a microscope that often infects the fur branch of the skin. Among the various species reported, at least Demodex folliculorum and Demodex brevis are found on the outer surface of the human body. The two Demodex species often coexist and preferentially gather in the same skin area of the face, cheeks, forehead, nose and ear canal, where active sebum secretion is beneficial to the environment and reproduction. Adult Demodex mites have a life cycle of 14 to 18 days and eventually die afterwards, and their proliferation depends on successful mating by the adult tick. Mating occurs at the opening of the hair follicle (near the outer surface of the skin). After this, the pregnant female moves to the sebaceous gland, lays eggs, and becomes a larva and a first nymph. The first nymph goes to the opening of the hair follicle and becomes the second nymph, crawls out of the skin and reenters the hair follicle to become an adult. Thus, if the adult Demodex can successfully mate during the life cycle and produce the next generation, the degree of infection in the host increases. Demodex infection does not appear in healthy children, but increases in an age-dependent manner, and is found in almost 100% of the skin of middle-aged people unless eradication is taken. The incidence of demodex infection is higher in unhealthy skin than in normal skin. The excessive growth of these ticks is associated with skin diseases and hair loss. Demodex is prevalent in limb mammals, particularly domestic animals, particularly dogs, and can cause follicular tick infections. Demodex enters the hair follicles and the sebaceous glands of animals, often causing severe dermatitis, infection and discomfort. On the other hand, Demodex is a mite that is parasitic in the hair follicles and sebaceous glands of the skin. As few as a few to as many as dozens of them inhabit each pore, and can reproduce by feeding fat in the sebaceous gland.

With reference to Examples 5 and 6, it was confirmed that the composition could inhibit or reduce Demodex survival.

According to an embodiment of the present invention, the composition may be a cosmetic composition. The cosmetic composition of the present invention contains the bacteriophage Pacnes 2012-15 as an active ingredient, and a basic cosmetic composition with a dermatologically acceptable excipient (face wash such as lotion, cream, essence, cleansing foam and cleansing water, pack, body oil) , Color cosmetic composition (foundation, lipstick, mascara, makeup base), hair product composition (shampoo, conditioner, hair conditioner, hair gel), and soap. The excipient is not limited thereto, but may include, for example, an emollient, a skin penetration enhancer, a colorant, a fragrance, an emulsifier, a thickener, and a solvent. In addition, flavoring, coloring, disinfecting agents, antioxidants, preservatives, moisturizing agents, etc. may be additionally included, and thickeners, inorganic salts, synthetic polymer materials, etc. may be included for the purpose of improving physical properties. For example, in the case of preparing a face wash and soap using the cosmetic composition of the present invention, it can be easily prepared by adding the composition of the present invention to a common face wash and soap base. In the case of preparing a cream, it can be prepared by adding the composition of the present invention to a general oil-in-water (O/W) cream base. Synthetic or natural materials such as proteins, minerals, vitamins, etc. for the purpose of improving physical properties, such as fragrances, chelating agents, coloring agents, antioxidants, preservatives, etc., may be additionally added.

When the cosmetic composition is formulated as a solution or emulsion, a solvent, a solubilizing agent, or an emulsifying agent may be used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl benzoate, propylene glycol, 1,3-Butylglycol oil may be used, and in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan may be used.

When the cosmetic composition is formulated as a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, an ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracant, and the like can be used.

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily implement the present invention. However, the present invention may be implemented in various different forms and is not limited to the embodiments described herein.

Manufacturing Example 1. Preparation of plant extract

After adding 10 times the volume of propandiol or butylene glycol in a 2:2:1 composition ratio of green lake, ginkgo biloba, and figs, extraction was performed at 60°C for 2 hours in an ultrasonic extractor, and then immersed at 4°C for 24 to 72 hours. Thereafter, after primary filtration with a 250 mesh filter cloth, secondary filtration was performed using a 0.45 μm membrane filter.

Experimental Example 1. Acne bacteria ( Propionibacterium acnes ) Antibacterial effect

Paper disc diffusion assay was performed to test the antibacterial effect of the composition according to the present invention against acne bacteria. Acne bacteria were inoculated and smeared on a solid plate medium (Reinforced clostridial Medium) for Propionibacterium acnes . Then, paper discs treated with 50, 100, 150 µl of the mixed extract of Artemisia Annua Extract, Ginkgo Biloba leaf Extract, and Fig (Ficus Carica (Fig) Fruit Extract) were placed on top of it and cultured. As a positive control, 20 µl of ampicillin 50µg/ml stock solution was treated. After incubation for 47 to 72 hours, the antibacterial activity was confirmed by measuring the extent to which acne bacteria did not grow around the paper disc.

As a result of the experiment, when 50, 100, and 150 µl of the mixed extracts of Cheongho, Ginkgo biloba, and figs were treated, the range of antibacterial power was measured as 0.05mm, 0,22mm, 0,3mm, respectively, at the outermost part of the paper disc. Confirmed (Table 1).

Experimental Example 2. Acne bacteria ( Propionibacterium acnes ) Growth inhibition effect

In order to test the growth inhibition effect of the composition according to the present invention against acne bacteria, a Growth inhibition assay was performed.

Acne bacteria were inoculated in a liquid medium for Propionibacterium acnes (Actinomyces Broth), and a mixture extract of Artemisia Annua Extract, Ginkgo Biloba leaf Extract, and Ficus Carica (Fig) Fruit Extract was added to 0. , 2, 4, 6, 8, 10% were treated. After incubation for 24 hours, the OD (optical dencity) value was measured with an absorbance spectrophotometer (UV-VIS sepcetrophotometer, Lambda 265, Perkin elmer) to compare the ability to inhibit the growth of acne bacteria.

As a result of the experiment, when 0, 2, 4, 6, 8, 10% of the mixed extract of blueberry, ginkgo biloba, and fig was treated for 24 hours, 7.83, 18.70, 36.50, 42.14, 51.55% of acne bacteria growth was suppressed compared to the untreated group. It was confirmed (Fig. 2). [Table 2] shows the measured value (OD) for inhibiting the growth of acne bacteria according to the treatment of mixed extracts of Cheongho, Ginkgo biloba, and Fig. [Table 3] shows the growth inhibition of acne bacteria according to the treatment of mixed extracts of Cheongho, Ginkgo biloba, and figs. It shows the converted value (OD).

Experimental Example 3. Acne bacteria ( Propionibacterium acnes ) Growth inhibition effect (SEM)

In order to confirm the growth inhibitory effect of the composition according to the present invention against acne bacteria, SEM (Scanning Electron Microscope) was taken.

Acne bacteria were inoculated in a liquid medium for Propionibacterium acnes (Actinomyces Broth), and a mixture extract of Artemisia Annua Extract, Ginkgo Biloba leaf Extract, and Ficus Carica (Fig) Fruit Extract was added to 0. , 2, 4% were treated. After incubation for 24 hours, acne bacteria were photographed with a scanning electron microscope (XL-30 FEG, FEI).

As a result of the experiment, it was confirmed that the original form (bacillus) of acne bacteria was deformed compared to the untreated group when 0, 2, 4% of the mixed extract of blueberry, ginkgo leaf, and fig were treated for 24 hours (Fig. 3).

Experimental Example 4. Anti-inflammatory effect

In order to confirm the anti-inflammatory effect of the composition according to the present invention, NF-kB promoter luciferase assay was performed.

NF-kB Luciferase Reporter NIH3T3 Stable Cell was dispensed into a 6-well culture plate at 1.5×10 ⁵ cells/well and cultured for 24 hours. After that, the mixture extract of Artemisia Annua Extract, Ginkgo Biloba leaf Extract, and Fig (Ficus Carica (Fig) Fruit Extract) was pretreated for 24 hours, and then TNF-α (50 ng) /mL) and incubated for 8 hours. After harvesting the cultured cells, the cells were lysed and centrifuged by adding passive lysis buffer (Promega) to extract luciferase in the cells. After taking only the supernatant and dispensing it on a Black 96-well plate, luciferin (Promega) was added, and the degree of luminescence of luciferin was measured using a Microplate reader system (SpectraMax®i3x Multi-Mode Detection Platform; Molecular Devices). The measured luminescence level of luciferin was calculated using the BCA Protein Assay Kit (Thermo). NF-kB luciferase activity was converted to the degree of luciferin emission per certain protein, and as the measured value decreases, NF-kB Luciferase Reporter NIH3T3 Stable Cell has an inhibitory effect on NF-kB activity.

As a result of the experiment, it was confirmed that when 0, 0.1, 1, 2, 4% of the mixed extract of blueberry, ginkgo biloba, and fig were treated for 24 hours, they inhibited NF-kB activity by 7.23, 20.23, 36.46, and 41.32%, respectively, compared to the untreated group. I did. (Fig. 4).

Experimental Example 5. Demodex reduction effect

Demodex reduction effect of the composition according to the present invention was confirmed through the following experiment. In addition, by varying the ratio of the raw material mixing ratio of the composition according to the present invention as shown in Table 4 below, the effect of reducing Demodex was measured.

To quantify sebum and observe Demodex, a digital light microscope (Advanced Microscopy Group Inc., USA), a slide glass, a cover glass, mineral oil (Bio-Rad, USA), and tweezers were used. During the experiment, sebum extracted from the nose was collected, placed on a slide glass, covered with a cover glass, and photographed, and the sebum was quantified through ImageJsoftware. After dissolving by dropping 20 μL of oil on the slide glass where sebum was collected, the cover glass was covered and the presence or absence of Demodex was observed using a Digitallightmicroscope. After 100 subjects were divided into 10 groups of 10 subjects, 1 g of sebum was collected, the average value of the number of Demodex was performed, and the results are shown in Table 5 below.

Demodex is difficult to observe because it forms a transparent shape after dying at room temperature within a short period of time, so Demodex was read within a short period of time for observation in a living state.

Experimental Example 6. Demodex survival inhibition effect

The compositions of Examples and Comparative Examples in Experimental Example 5 were suspended in purified water and diluted, and the effect of inhibiting Demodex survival was evaluated.

After attaching the electron microscope to the CCD camera, it was connected to the computer. After separating the Demodex from the hair follicle using a blade, the extracts of Examples and Comparative Examples, which were placed on a slide glass and diluted to a concentration of 100 μg/mL, were treated. The cover glass was covered and the movements of the Demodex body and legs were observed under 3 electron microscopes. The electron microscope used a low magnification to observe the appearance of the Demodex, and then gradually observed the detailed appearance at a high magnification to check the time and measure the survival time. In the experiment, the survival time was measured from the time the sample was dropped to the time when the Demodex body gradually shrunk and the movement was terminated, and is shown in Table 6 below.

Referring to [Table 6], Examples 1 to 3 were excellent in inhibiting the survival of Demodex compared to Comparative Examples 1 to 4.

The above description of the present invention is for illustrative purposes only, and those of ordinary skill in the art to which the present invention pertains will be able to understand that other specific forms can be easily modified without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are illustrative and non-limiting in all respects. For example, each component described as a single type may be implemented in a distributed manner, and similarly, components described as being distributed may also be implemented in a combined form.

The scope of the present invention is indicated by the claims to be described later, and all changes or modified forms derived from the meaning and scope of the claims and their equivalent concepts should be interpreted as being included in the scope of the present invention.

Claims (7)

As an antibacterial and anti-inflammatory composition comprising blueberry extract, ginkgo leaf extract and fig extract as active ingredients,
The composition is characterized in that it has an inhibitory function against acne bacteria (Propionibacterium acnes), antibacterial and anti-inflammatory composition. The method of claim 1,
The composition is an antibacterial and anti-inflammatory composition comprising 20 to 40 parts by weight of the blueberry extract, 20 to 40 parts by weight of the ginkgo leaf extract, and 10 to 20 parts by weight of the fig extract. The method of claim 1,
The extract is composed of water, anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms, acetone, petroleum ether, ethyl acetate, butyl acetate, trichloromethane, dichloromethane, chloroform, hexane and 1,3-butylene glycol, propanediol Antibacterial and anti-inflammatory composition, characterized in that extracted with one or more solvents selected from the group. The method of claim 3,
The extract is an antibacterial and anti-inflammatory composition, characterized in that extracted by ultrasonic extraction in the solvent. delete The method of claim 1,
The composition is an antibacterial and anti-inflammatory composition, characterized in that it has a function of inhibiting NF-kB activity. The method of claim 1,
The composition is an antibacterial and anti-inflammatory composition, characterized in that the cosmetic composition.

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