KR20220104655A - Composition for treating acne containing nanoized bee pollen extract as an active ingredient - Google Patents

Abstract

The present invention relates to a composition for the treatment of acne containing a nanonized bee pollen extract as an active ingredient. More particularly, the present invention provides a composition for treating acne containing a nanonized bee pollen extract as an active ingredient which has a clear acne alleviation effect and enables long-term administration by a combination of nanonized bee pollen and purified bee venom with excellent anti-inflammatory efficacy.

Description

Composition for treating acne containing nanoized bee pollen extract as an active ingredient}

The present invention relates to a composition for treating acne derived from a natural product, and more particularly, to a composition for treating acne comprising a nanoized bee pollen extract as an active ingredient.

Acne is a chronic inflammatory disease of the sebaceous glands that is generated due to retention hyperkeratosis in the pores due to the clogging of sebum. Rather, it can be said that it is one of the characteristics of the skin during the growth period. The causes of acne can be largely explained according to the degree of development of acne symptoms, such as increased sebum secretion, stenosis by hyperkeratosis of hair follicles, proliferation of acne bacteria, and inflammatory response. It may act in combination. Excessive secretion of sebum, keratinization of hair follicles, and proliferation of acne bacteria are three major factors that cause acne, but hypertrophy of sebaceous glands, hyperkeratosis of hair follicles, and bacterial influences act in a complex way, exacerbating acne. and each one alone creates acne. In addition, genetic factors that are prone to acne, external factors such as season and climate, influence of food, fatigue and stress are also related to acne. Although the cause of acne is still not clearly identified, it has been confirmed through research reports so far that the three important causes are directly involved in the onset of acne if genetic and environmental factors are excluded. In order to treat such acne, there is a method of using an oral or external drug form. However, in the case of the therapeutic agent, there are side effects such as hormone abuse, skin irritation and contact dermatitis, so the need for the development of products that are safe and effective for the human body is constantly emerging. In this regard, Korean Patent No. 1459742 discloses a cosmetic composition for improving acne containing a natural extract.

However, in the case of the prior art, there has been no research on a composition for treating acne comprising purified bee venom and nano-sized bee pollen as active ingredients, which contains extracts of chrysanthemum chrysanthemum, Chrysanthemum and cinnamon as active ingredients.

The present invention is to solve various problems including the above problems, and it is a combination of purified bee venom, which has excellent anti-inflammatory effect, and nanoized bee pollen. An object of the present invention is to provide a composition for treating acne comprising: However, these problems are exemplary, and the scope of the present invention is not limited thereto.

According to one aspect of the present invention, there is provided a composition for treating acne, comprising a nano-sized bee pollen composition as an active ingredient and a tablet bee venom, and a pharmaceutically acceptable carrier.

According to another aspect of the present invention, there is provided an external preparation for acne treatment comprising the composition.

According to another aspect of the present invention, there is provided a cosmetic composition for improving acne, comprising the composition.

The composition for the treatment of acne comprising the nanoized bee pollen extract of the present invention made as described above as an active ingredient combines the significant effects of nanoized bee pollen derived from natural products and bee venom to provide excellent acne treatment effects according to anti-inflammatory effects. can be implemented Therefore, it can be applied to product groups related to acne skin treatment and all industrial fields. Of course, the scope of the present invention is not limited by these effects.

1 is a graph analyzing the NO production inhibitory effect according to the treatment of the composition of the present invention.
Figure 2 is a graph analyzing the change in the pro-inflammatory cytokine TNF-α content according to the treatment of the composition of the present invention.
Figure 3 is a graph analyzing the effect on the pro-inflammatory cytokine IL-6 profile according to the treatment of the composition of the present invention.
4 is a graph analyzing the inhibitory effect of β-hexaminidase activity according to the treatment of the composition of the present invention.
5 is a graph analyzing the leukotriene (LT) production inhibitory effect according to the treatment of the composition of the present invention.

Definition of Terms:

The term "bee pollen" used in this document is a mixture of nectar and nectar when worker bees collect nectar and pollen. The bee pollen is known as a nutritional substance containing various minerals, vitamins, and essential amino acids, and many European athletes are taking the bee pollen collected by a collector for a long time as a nutritional supplement to improve athletic performance.

The term "Bee Venom" used in this document is contained in the venom sac of bees, and is composed of about 40 substances and has been used to treat diseases of the human body since BC. The composition of bee venom differs depending on the type of bee, and even the same bee has a difference in composition according to the age of the bee. Currently, studies are being reported on the action and mechanism of bee venom components, hypersensitivity and toxicity of bee venom, immunotherapy and treatment of diseases such as arthritis, herpes simplex, multiple sclerosis, and tumors. is known Domestic bee venom ( Apis mellifera ) can be obtained by using a bee venom collecting device using an electric shock method to collect only pure bee venom in large quantities without damaging the bees, remove foreign substances and freeze-drying to obtain purified bee venom.

DETAILED DESCRIPTION OF THE INVENTION:

According to one aspect of the present invention, there is provided a composition for treating acne, comprising a nano-sized bee pollen composition as an active ingredient and a tablet bee venom and a pharmaceutically acceptable carrier.

In the composition, 1 to 20% by weight of the nanoized bee pollen composition, and 0.000001 to 1% by weight of purified bee venom, and 0.1 to 4% by weight of a dispersant, 0.1 to 4% by weight of a preservative, 1 to 10% by weight of a thickener , 0.05 to 0.3% of antioxidant and 0.1 to 1% by weight of neutralizing agent, and the remainder may be mixed to consist of distilled water.

In the composition, the nanoized bee pollen may be prepared by a wet nano-pulverization process after mixing distilled water with the bee pollen, and the nanosized bee pollen may have a particle size of 50 to 500 nm, and the nanosized bee pollen may be The exine may be removed, and the bee venom may be isolated from the venom sac of a worker bee over 15 days of age of a Western species of honey bee ( Apis mellifera L.).

According to another aspect of the present invention, there is provided an external preparation for acne treatment comprising the composition.

According to another aspect of the present invention, there is provided a cosmetic composition for improving acne, comprising the composition.

The composition of the present invention can be applied in any shape, such as liquid, cream, paste or solid, and includes not only creams, skin lotions, packs, and makeup bases, but also face wash such as soap, facial foam, body cleanser, detergent and shampoo; It can also be applied to hair products such as conditioners, and if necessary, it is also possible to mix with ingredients commonly used in external preparations for skin, for example, water-based ingredients, oil-based ingredients, surfactants, moisturizers, thickeners, preservatives, and fragrance pigments. . In addition, it can be prepared and applied in the form of powder, granules, capsules, or injections by mixing with a pharmaceutically acceptable carrier, forming agent, dilute, etc. It can also be applied as a cleaning tool.

The composition of the present invention can exhibit excellent effects beyond the effects of each component in the prevention or treatment of acne due to the effects of nano-sized bee pollen and purified bee venom. In the composition of the present invention, it is preferable that 1 to 20% by weight of the nanoized bee pollen and 0.00001 to 1% by weight of the purified bee venom of the composition are added in order to exert an effect of preventing or treating acne, and in the case of bee venom, 0.0001 to 0.5% by weight is more preferred. When the content of the purified bee venom is less than 0.00001 parts by weight, it is difficult to obtain the acne treatment effect, which is the desired effect of the present invention.

The composition of the present invention further comprises any one or more components selected from the group consisting of green tea extract, collagen, moisturetin, grape seed oil, licorice extract, allantoin, soybean oil and tocopherol, and when these components are additionally contained, acne improvement effect can be improved. Each of these components is preferably included in an amount of 1 to 20% by weight, respectively, based on the total weight of the composition. However, there is no significant limitation in weight.

The composition of the present invention is based on the composition of Patent No. 10-1572067, which has already been evaluated for antiseptic effect, application effect, cytotoxicity, and stability in order to maximize the effects of nanosized bee pollen and bee venom, 1 to 20% by weight of nanoized bee pollen , purified bee venom, 0.000001 to 1% by weight, and distilled water for the remainder to prepare a composition for treating acne, dispersant 0.1 to 4% by weight, preservative 0.1 to 4% by weight, thickener 1 to 10% by weight, antioxidant 0.05 to 0.3% and 0.1 to 1% by weight of a neutralizing agent may be additionally included and the remainder may be configured by mixing to constitute a distilled water.

The composition of the present invention can be applied in any shape such as liquid, cream, paste, or solid, and includes not only creams, skin lotions, packs, and makeup bases, but also cleansing agents such as soap, facial foam, and body cleanser, and It can be applied to hair products such as shampoo and conditioner, and it can be in the form of a patch such as a mask pack. In addition, if necessary, it is possible to mix it with components commonly used in external preparations for skin, for example, water-based components, oil-based components, surfactants, moisturizers, thickeners, preservatives, fragrance pigments, and the like.

In the composition of the present invention, a natural preservative MSE (Multi-Scutellaria Etracts) may be used as a preservative. MSE is extracted from the root of gold, a plant of the Lamiaceae family, and has antibacterial effects on various skin inflammations and skin troubles, and helps with antioxidant, whitening and anti-aging effects. In addition, the composition of the present invention may be prepared as a semi-solid preparation such as a cream, lotion, emulsifier, liniment agent, etc., but is not limited thereto, and a suitable colorant, flavoring agent, stabilizer, viscous agent, surfactant, etc. It can be prepared by adding In addition, the composition for external application for skin according to the present invention may be prepared as a liquid formulation and includes, but is not limited to, a solution such as water, alcohol, or propylene glycol solution, a suspension, an emulsion, and the like, and a suitable colorant, flavoring agent, and stabilizer. , a viscous agent, etc. may be added. Specifically, the external preparation for skin includes a suitable pharmaceutically acceptable base such as vaseline and stearyl alcohol; suitable pharmaceutically acceptable surfactants such as polysorbate and sorbitan sesquioleate; Suitable pharmaceutically acceptable moisturizers such as glycerin; a suitable pharmaceutically acceptable solvent; And it may be prepared by a conventional skin external preparation method of homogeneously mixing a flavoring agent, a colorant, a stabilizer, a viscous agent, and the like.

The composition of the present invention may be administered in an amount of about 0.0001 mg/cm ² of skin area to about 5 g/cm ² of skin area every day, and daily administration of about 0.001 mg/cm ² of skin area to about 1 g/cm ² of skin area capacity is preferred. However, the dosage may vary depending on the route of administration and formulation, the patient's condition (age, sex, weight, etc.), and the severity of the condition being treated. For convenience, if necessary, the total daily dose may be divided and administered several times during the day.

To date, there is a method of using an oral or external drug form to treat acne. Specifically, in addition to the method of inhibiting sebum production using anti-male hormones, and treatment by the anti-inflammatory action of steroid hormones and non-steroidal anti-inflammatory drugs, resorcinol, sulfur, salicylic acid, benzoyl peroxide, retinoic acid or isotretinoin There is a method of inhibiting the activity of Propionibacterium acnes by antibiotics such as antibacterial agents and exfoliants, tetracycline, erythromycin, and macrocycline. Meanwhile, recently, treatment methods using azelaic acid have been widely used. However, the methods described above have some problems in terms of effectiveness, side effects, and usability, although achieving some degree of performance in the treatment of acne. The use of hormones has been reported to have side effects of hormone abuse such as inhibition of epidermal growth, and exfoliants such as retinoic acid and benzoyl peroxide have concerns about skin irritation and contact dermatitis due to keratin exfoliation, and antibiotics such as tetracycline The substance is known to cause bleeding of resistant bacteria and photosensitivity. In particular, isotretinoin is known to have a teratogenic effect on the fetus. Therefore, the need for the development of products that are stable to the human body and have excellent effects for acne-prone skin is constantly emerging. On the other hand, purified bee venom has been proven to have anti-inflammatory and antibacterial effects, and has high potential for development as a treatment for acne bacteria and is being studied. In this regard, Korea Registered Patent No. 10-1459742 for acne improvement cosmetic composition containing extracts of natural products, and Korean Registered Patent No. 10-1835397 Inflammation and allergic diseases containing a combination extract of Nahanpoja, Nahan, and coniferous herb as an active ingredient. Although there have been many studies such as preventive and therapeutic compositions, the composition for improving acne mainly made from herbal medicine compositions and natural material compositions is complicated by the manufacturing process because there are many processes and substances contained therein. Therefore, the inventors of the present inventors have removed the allergen component by crushing the outer shell of the nano-sized bee pollen, and the extract has excellent antibacterial, anti-inflammatory, and antiviral power, and thus the value is recognized as a cosmetic composition. Based on this, a composition for treating acne containing two natural materials was completed. Although purified bee venom and nanoized bee pollen have excellent antibacterial activity and cosmetic composition, they have not yet been used as a composition limited to acne, so it is considered to be suitable as a convergence material. Therefore, in the present invention, the composition containing melatin of purified bee venom and nanoized bee pollen in keratinocytes and monocytes induced in inflammation caused by acne bacteria suppresses the inflammatory response through regulation of inflammation and apoptosis, and has antibacterial activity with commercially available drugs. The excellent anti-inflammatory effects of bee venom and bee pollen were confirmed through comparative tests. In addition, the safety, antigenicity, histamine test, phototoxicity, photosensitivity, and skin irritation evaluation to confirm the safety for improving acne, the composition of the present invention will be useful as various cosmetic preparations and therapeutic preparations.

The present invention relates to a composition for improving and treating acne prepared for the purpose of treating anti-inflammatory and allergic symptoms, using two naturally-derived complex extracts as an active ingredient as a convergence bio material of purified bee venom and nanoized bee pollen. Its astrological properties are: 1) LPS stimulation represses the NO, TNF-a, and IL-6 cytokine production mechanisms secreted by macrophages; 2) β-Hex release and secreted by mast cells; By also inhibiting the LTC4 production mechanism, the production of IgE, IL-4, IL-5 and IL-13 in the bronchoalveolar lavage fluid of an animal model of asthma caused by ovalbumin was inhibited, and IL-4, IL-5 in the lungs was inhibited. and IL-13, which significantly exhibits anti-inflammatory and anti-allergic effects, and thus can be used for the prevention and treatment of inflammatory and allergic diseases as a complex extract of two natural substances.

In relation to the composition, researches such as extraction of specialized ingredients from the corresponding substances are active, but technology development focusing on the skin cleansing effect opens closed pores while minimizing skin irritation with the exfoliating effect of cleansing and moisturizing ingredients. Enzyme development technology to generate free fatty acids by suppressing excess secretion of male hormones with the effect of controlling excess sebum, technology to prevent acne by removing excess sebum by fine particles that can adsorb sebum, antibacterial As for the effect, it is a technology to prevent acne by performing antibacterial action with a broad spectrum such as Propionbacterium ancness and various bacteria and fungi harmful to the skin, and PDT (photodynamic therapy) such as ALA, MAL, etc. Acne treatment technology is being developed through In this regard, the purified bee venom of the present invention and nanoized bee pollen have excellent antibacterial activity as a cosmetic composition, but have not yet been used as a composition limited to acne, so it is considered to be suitable as a convergence material. In addition, after the development of the bee venom collecting device, domestic bee venom production has been actively carried out. These purified bee venom have proven anti-inflammatory and antibacterial effects, and are highly likely to be developed as a treatment for acne bacteria.

Hereinafter, the present invention will be described in more detail through examples. However, the present invention is not limited to the embodiments disclosed below, but can be implemented in a variety of different forms. It is provided to fully inform

experimental material

The bee pollen used in the present invention was purchased from a bee pollen farm in Namwon, Jeollabuk-do and Sancheong, Gyeongsangnam-do, and cell culture medium RPMI-1640, MEM (Modifed Eagle Medium), non-essential amino acids solution, FBS ( Cell culture reagents such as fetal bovine serum), streptomycin, and penicillin were purchased from Hyclone (Logan, UT, USA). Also, among the reagents used in the experiment, SCF (Stem cell factor) was STEMCELL (Vancouver, Canada), LPS (Lipopolysaccharides) and ovalbumin (Albumin, Albumin from chickenegg white) were Sigma (St. Louis, MO, USA), LTC4 measurement kit is Cayman (Ann Arbor, MI, USA), alum is Thermo (Hanover Park, IL, USA), IgE measurement product is BD Biosciences (San Jose, CA, USA), for IL measurement The kit was purchased from R&D Systems (Minneapolis, MN, USA) and used.

Preparation of wet nano-grinded bee pollen

The wet nano-grinding process of bee pollen was performed using a wet mill (Rotate Mill; Ilsin Nanotechnology, Japan) through the prior art (Choi et al., Journal of Life Science. 26(6): 651-656, 2016). However, specifically, a bee pollen mixture was prepared by mixing 10 g of bee pollen per 100 ml of distilled water, and the mixture was wet-pulverized for 3 hours. Thereafter, the particle size distribution of the wet-pulverized bee pollen was confirmed using a particle size analyzer (Mastersizer 2000; Malvern Instruments, UK), and as a result, it was confirmed that most were less than or equal to 500 nm. 300 g of nanoized bee pollen was obtained.

Tablet Bee Venom Manufacturing

The present inventors isolated bee venom from the venom sac of Western bee ( Apis mellifera L.) worker bees, and collected pure bee venom in large quantities without harm to the bees by using a bee venom collecting device through an electric shock method. Thereafter, foreign substances were removed from the bee venom, and the collected bee venom was dissolved in distilled water, centrifuged, filtered, and then freeze-dried to obtain 25 μg of pure purified bee venom.

Preparation of mixtures for cell experiments

The freeze-dried nanoized bee pollen obtained according to an embodiment of the present invention was mixed with distilled water at concentrations of 3 µg/mL, 4 µg/mL, 5 µg/mL and 10 µg/mL, and bee venom was 2 ng/ mL was added.

Preparation of mixtures for skin testing

A skin test mixture was prepared according to an embodiment of the present invention. Specifically, 1 ml of a bee venom solution prepared by adding 0.1 g of purified bee venom to 1 ml of distilled water and 0.5 g of the nanoized bee pollen prepared above were used in patent No. 10-1572067, whose preservative effect, application effect, cytotoxicity and stability were already evaluated. 99 ml of a mixture consisting of 0.1 to 4% by weight of a dispersant prepared based on the composition, 0.1 to 4% by weight of a preservative, 1 to 10% by weight of a thickener, 0.05 to 0.3% of an antioxidant and 0.1 to 1% by weight of a neutralizing agent, with the remainder being distilled water After adding to the mixture, it was stirred to prepare a composition for external application for skin treatment for acne.

cell culture

RAW 264.7 cells, a mouse macrophage cell line used in the present invention, were cultured in DMEM medium containing 10% FBS, 100 U/mL penicillin, and 100 μg/mL streptomycin at 37° C. and 5% CO ₂ conditions. In addition, mouse bone marrow-derived mast cells (BMMC) were isolated from bone marrow cells from the femur of BALB/C mice, containing 10% FBS, 100 U/mL penicillin and 100 μg/mL streptomycin. In RPMI-1640 medium, PWM-SCM (containing pokeweed mitogen-stimulated spleen cell-conditioned medium) as an IL-3 source is added to a final concentration of 20%, and cultured for about 3 weeks to obtain 90% or more homogeneous BMMC. did. The cells were cultured in an incubator at 37° C., 5% CO ₂ , and a constant number of BMMC cells were dispensed in a 96-well plate.

Experimental Example 1: NO (nitric oxide) production inhibitory effect

The present inventors confirmed the NO production inhibitory effect by treating RAW 264.7 cells with the composition of the present invention. Specifically, the experiment was performed as follows using the method described in the existing literature (Yang, JH et al., Immunopharmacol Immunotoxicol. 33(2):373-379, 2011). First, RAW 264.7 cells (1.5×10 ⁵ cells/well) were pre-cultured in a 24-well plate and replaced with DMEM medium. After pretreatment with the composition of the present invention for 1 hour at each concentration, 200 ng/mL of LPS was added to each well. Treated and incubated for 24 hours at 37 ℃, 5% CO ₂ conditions. After that, the supernatant was collected and reacted by mixing the same amount of Griess reagent (G4410, Sigma) in 100 μL, and then the absorbance at 570 nm was measured with an ELISA reader (Sunrise-Basic Tecan, TECAN). The amount of NO production was analyzed using a standard curve for each concentration of sodium nitrite.

As a result, when the composition of the present invention was treated at concentrations of 4 μg / mL, 5 μg / mL and 10 μg / mL, the production of nitrite was inhibited to 11 μM, 9 μM and 4 μM, respectively (Fig. One).

Experimental Example 2: Inhibitory effect of pro-inflammatory cytokine production

The present inventors performed the inhibitory effect on the production of pro-inflammatory cytokines TNF-α and IL-6 according to the treatment of the composition of the present invention with reference to prior research literature (Yang JH et al., Immunopharmacol Immunotoxicol, 33(2): 373-379, 2011). Specifically, RAW 264.7 cells (1.5×10 ⁵ cells/well) were pre-cultured in a 24-well plate and replaced with DMEM medium. The cells were pre-treated with the composition of the present invention for 1 hour, and LPS 200 ng/ml was added to each well. After treatment in 37 ℃, 5% CO ₂ conditions were incubated for 24 hours. Then, the supernatant was collected and the amount of pro-inflammatory cytokines TNF-α and IL-6 was measured using an ELISA kit (R&D System, USA).

As a result, in the LPS-treated cell supernatant, the production of TNF-α was significantly increased to 114 ng/mL, but the composition of the present invention was pre-treated at concentrations of 4 μg/mL, 5 μg/mL and 10 μg/mL. In one case, TNF-α production was shown to decrease in a concentration-dependent manner to 80 ng/mL, 76 ng/mL and 68 ng/mL (FIG. 2). In addition, in the LPS-treated cell supernatant, the production of IL-6 was significantly increased to 1125 pg/mL, but as a result of pre-treatment with the composition of the present invention at concentrations of 4 μg/mL, 5 μg/mL and 10 μg/mL It was also shown to decrease in a concentration-dependent manner to 601 pg/mL, 503 pg/mL and 450 pg/mL (FIG. 3). The above results suggest that the composition of the present invention exhibits excellent anti-inflammatory activity.

Experimental Example 2: β-Hex release inhibitory effect

The present inventors performed the inhibitory effect on the release of β-hexaminidase (β-Hex), which is a marker of degranulation of BMMC, according to the treatment of the composition of the present invention with reference to the prior research literature (Son JK et al . , Biol Pharm Bull, 28(12):2181-84, 2005). Specifically, 2×10 ⁵ cells/well of BMMC was treated with the composition of the present invention at different concentrations (3 to 5 μg/mL) and pre-cultured for 30 minutes at 37° C., 5% CO ₂ conditions, and then SCF (stem cell factor) (100 ng/mL) (STEMCELL Technologies, Vancouver, Canada), incubated for 15 minutes, and centrifuged at 3000 rpm, 4°C for 5 minutes. Thereafter, the supernatant was mixed with β-Hex substrate {100 mM citrate buffer (citric acid 0.955%, sodium citrate dihydrate 1.478%, pH 4.5), 1.3 mg/mL p-nitrophenyl-N-acetyl-bD- After mixing with glucosaminide (p-nitrophenyl-N-acetyl-bD-glucosaminide) in a mixing ratio of 1:2 (w/v) and reacting at 37°C for 1 hour, 0.2M glycine (pH 10.7) (Sigma) to stop the reaction, the absorbance at 405 nm wavelength was measured using an ELISA reader (Tecan System, San Jose, CA, USA), and the measured value was converted into an inhibition ratio (inhibition %).

As a result, it was shown that the composition of the present invention inhibited the release of β-hexaminidase by 25%, 30% and 43%, respectively, at treatment concentrations of 3 μg/mL, 4 μg/mL and 5 μg/mL (FIG. 4). ).

Experimental Example 3: Leukotriene production inhibitory effect

The present inventors performed the inhibitory effect of leukotriene (LT) production according to the treatment of the composition of the present invention with reference to prior research literature (Hua JM et al., Arch Pharm Res , 31:617-621, 2008). Specifically, in order to confirm LT production, 1 × 10 ⁵ cells/mL BMMC was treated with the composition of the present invention at different concentrations (3 to 5 μg/mL) and cultured for 1 hour at 37° C., 5% CO ₂ conditions, and After stimulation with SCF (stem cell factor) at a concentration of 100 ng/mL for 15 minutes, the supernatant was separated by a centrifuge (Centrifuge 5415R, Eppendorf) at 3000 rpm, 4° C. for 5 minutes. Then, the LTC ₄ content in the supernatant was measured using the LTC ₄ -EIA kit (Cayman). As a result, it was found that the composition of the present invention inhibited the production of LTC ₄ by 30%, 51% and 90%, respectively, at the treatment concentrations of 3 μg/mL, 4 μg/mL and 5 μg/mL ( FIG. 5 ).

Experimental Example 4: Antibacterial activity against multidrug-resistant bacteria

When infected with multidrug-resistant bacteria (MRSA), the symptoms of skin and soft tissue infection are like a bump on the skin, causing general swelling, pus, and pain. In addition, the infection site appears as acne, weak burns, or insect bites, and the present inventors performed an antibacterial activity according to the treatment of the composition of the present invention as a complementary experiment with reference to the prior research literature (Kwak, MK et al., Food Control 85: 223-234, 2018). Specifically, the antibacterial activity against the multidrug-resistant strains Staphylococcus aureus 11471, Streptococcus pneumoniae 14596, and Salmonella typhimurium 12219 was determined by comparing the minimum inhibitory concentration (MIC) to obtain the minimum concentration and the active concentration against the fungus as human opportunistic/pathogenic fungi. Candida albicans (Candida albicans) and the phytopathogenic fungus Ganoderma boinense (Ganoderma boninense) were confirmed in each 6-well. As a result, it was found that the antibacterial and antifungal activity of the mixture containing the nanoized bee pollen and bee venom of the present invention containing a naturally-derived component against multidrug-resistant bacteria was high. Antimicrobial activity evaluation results for the composition of the present invention against the multi-drug-resistant bacteria are summarized in Table 1 below.

antibacterial activity Multidrug-resistant strains Active concentration of bee pollen-bee venom mixture (MIC, μg/L)* Staphylococcus aureus 114711 7.3 ± 0.77 Streptococcus pneumoniae 145961 8.5 ± 0.32 Salmonella typhimurium 122192 5.1 ± 0.54 fungus Active concentration of the mixture (μg complex in 3 mL agar assay)* plant pathogen Ganoderma boninense 110.9 ± 3.56 human pathogen Candida albicans 101.4 ± 3.26

*Data represent ± SD of 3 independent experiments expressed in mg/L of each test sample.

Experimental Example 5: Formulation stability evaluation

The present inventors stored the composition of the present invention at a temperature of 40 ° C, room temperature (25 ° C), -10 ° C. for 25 days to experimentally evaluate the degree of stabilization, and as a result of confirming phase safety, the formulation was changed under all the temperature conditions appeared to be absent.

Experimental Example 6: Measurement of acne treatment effect

The present inventors prepared an external preparation containing the composition of the present invention and observed the acne treatment effect. Specifically, the skin test mixture was prepared according to the above-described manufacturing method and then stored in a refrigerator. After that, for 17-20 year old female students with acne skin, 7 students per group, total of 6 groups, for a total of 42 people, once a day for 7 days, after washing the face in the evening, apply the topical agent containing the composition of the present invention to the entire face. It was applied in an appropriate amount, and the acne treatment effect was observed for 7 days.

As evaluation criteria, if the acne treatment effect is significant, that is, if the acne inflammation level is clearly reduced, 5 points, if the inflammation level is improved, 4 points, if the inflammation level remains but improvement is seen, 3 points, if there is no change, 2 points, and In the case of more aggravation, the total score was given as 1 point, and overall judgment was made. As a comparison group, distilled water without bee venom and bee pollen and distilled water containing 5% alcohol was used.

As a result, the excellent acne treatment effect of the experimental group to which the external preparation containing the composition of the present invention was applied was confirmed. The measurement results of the acne treatment effect are summarized in Table 2 below.

Subject 1 Subject 2 Subject 3 Subject 4 Subject 5 Subject 6 Subject 7 total score composition 5 4 5 5 4 4 5 32 Distilled water One 2 2 One 3 2 One 12 alcoholic distilled water 3 2 One 3 2 2 One 14

Experimental Example 7: Antibacterial activity evaluation experiment

The present inventors performed an antibacterial activity evaluation experiment of the external application containing the composition of the present invention using Propionibacterium acnes , which is an acne bacterium as a test bacteria. Specifically, each of the acne bacteria was inoculated into BH1 broth and pre-cultured, and then the composition of the present invention was inoculated at a concentration of 1% in 100 ml of sterilized fresh BHI broth containing 5%, and then cultured in an anaerobic incubator for 2 days. was measured. The source of the strain is involved in sebum secretion for acne and abnormal keratinization of hair follicles to form a bacterial community of the strain, so the growth of the strain was observed when relieving localized chronic inflammatory disease caused by an inflammatory reaction.

As a result, it was confirmed that the growth of acne bacteria was significantly reduced according to the treatment of the composition of the present invention. The above results suggest the excellent antibacterial activity of the composition of the present invention. The antibacterial activity evaluation test results are summarized in Table 3 below.

Antibacterial activity evaluation test result division Acne ( Propionibacterium acnes ) Inoculum concentration (cfu/mL) Bacteria concentration after culture (cfu/mL) Example 1 2.32 × 10 ⁶ 1.3 × 10 ² Comparative Example 1 2.32 × 10 ⁶ 2.0 × 10 ⁵

In conclusion, the composition for treating acne comprising the nano-sized bee pollen extract of the present invention as an active ingredient contains as active ingredients Bee Pollen wet-pulverized into nano-sized particles and purified bee venom to provide anti-inflammatory, antibacterial, and anti-inflammatory properties. Since the virus effect is excellent, it can be used as a pharmaceutical composition or a food composition for improving skin diseases such as acne treatment, prevention and treatment of inflammation and allergic diseases.

Although the present invention has been described with reference to the above-described embodiments, these are merely exemplary, and those skilled in the art will understand that various modifications and equivalent other embodiments are possible therefrom. Therefore, the true technical protection scope of the present invention should be determined by the technical spirit of the appended claims.

Claims (9)

A composition for treating acne, comprising a nanosized bee pollen composition as an active ingredient and a tablet bee venom, and a pharmaceutically acceptable carrier. According to claim 1,
A composition comprising 1 to 20% by weight of a nanosized bee pollen composition, and 0.000001 to 1% by weight of purified bee venom. According to claim 1,
A composition further comprising 0.1 to 4% by weight of a dispersant, 0.1 to 4% by weight of a preservative, 1 to 10% by weight of a thickener, 0.05 to 0.3% of an antioxidant and 0.1 to 1% by weight of a neutralizing agent. According to claim 1,
The nanoized bee pollen is prepared by mixing distilled water with the bee pollen and then using a wet nano pulverization process. According to claim 1,
wherein the nanosized bee pollen has a particle size of 100 to 500 nm. According to claim 1,
The nanoized bee pollen has an outer shell (exine) removed, composition. According to claim 1,
The bee venom is isolated from the venom sac of worker bees over 15 days of age of Western honey bees ( Apis mellifera L.). Claims 1 to 7, comprising the composition of any one of claims, for external use for acne treatment. A cosmetic composition for improving acne, comprising the composition of any one of claims 1 to 7.

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