KR102210621B1 - Composition for improving skin wrinkle and skin moisturizing comprising Pediococcus inopinatus as effective component - Google Patents
Composition for improving skin wrinkle and skin moisturizing comprising Pediococcus inopinatus as effective component Download PDFInfo
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- KR102210621B1 KR102210621B1 KR1020190054271A KR20190054271A KR102210621B1 KR 102210621 B1 KR102210621 B1 KR 102210621B1 KR 1020190054271 A KR1020190054271 A KR 1020190054271A KR 20190054271 A KR20190054271 A KR 20190054271A KR 102210621 B1 KR102210621 B1 KR 102210621B1
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Abstract
본 발명은 페디오코쿠스 이노피나투스를 유효성분으로 포함하는 피부 주름 및 피부 보습의 예방, 개선 또는 치료용 조성물에 관한 것이다. 상세하게는 페디오코쿠스 이노피나투스는 자외선 조사에 의해 증진된 MMP-1 및 MMP-9의 발현을 현저하게 억제하고, 피부 각질층을 완화하여 피부두께를 감소시키며, 피부 수분량을 증진시키고, 경피수분 손실량을 감소시키며, 히알루론산의 함량을 증진시키는 것이다. 따라서 본 발명의 조성물은 피부 주름 개선 또는 피부 보습용 화장료 조성물; 피부주름 개선 또는 피부 보습용 건강기능식품 조성물; 피부 건조증의 예방 또는 치료용 약학 조성물; 피부주름 개선 또는 피부 보습용 사료 조성물; 및 피부주름 개선 또는 피부 보습용 수의학적 조성물로 사용할 수 있다.The present invention relates to a composition for preventing, improving or treating skin wrinkles and skin moisturizing, comprising Pediococcus inofinatus as an active ingredient. Specifically, Pediococcus Inofinatus remarkably inhibits the expression of MMP-1 and MMP-9 enhanced by UV irradiation, relieves the stratum corneum, reduces skin thickness, improves skin moisture, and improves transdermal moisture. It reduces the amount of loss and increases the content of hyaluronic acid. Therefore, the composition of the present invention is a cosmetic composition for improving skin wrinkles or moisturizing the skin; Skin wrinkle improvement or skin moisturizing health functional food composition; A pharmaceutical composition for preventing or treating dry skin; Skin wrinkle improvement or skin moisturizing feed composition; And it can be used as a veterinary composition for improving skin wrinkles or moisturizing the skin.
Description
본 발명은 페디오코쿠스 이노피나투스를 유효성분으로 포함하는 피부 주름 개선 및 피부 보습용 조성물에 관한 것이다.The present invention relates to a composition for improving skin wrinkles and moisturizing skin, comprising Pediococcus inofinatus as an active ingredient.
피부는 외부의 환경과 항상 접하고 있는 기관으로 주로 외부의 물리적 손상 및 화학물질로부터 인체를 보호하고, 세균, 곰팡이, 바이러스 등이 피부로 침범하는 것을 방지하며, 수분 손실을 막는 보호장벽 역할을 한다. 피부의 구조를 보면 표피, 진피, 피하지방의 3개 층으로 구성되고 표피는 3개 층 중 가장 얇은 층으로 피부의 보습 및 보호를 담당하는 중요한 기능을 한다. 특히 표피의 각질층에 존재하는 수분에 의하여 피부가 탄력 있고 부드럽게 되는데, 각질층의 탄력이 유지되려면 10% 이상의 수분함유가 필수적인 것으로 알려져 있다.The skin is an organ that is always in contact with the external environment. It mainly protects the human body from external physical damage and chemicals, prevents bacteria, fungi, and viruses from invading the skin, and acts as a protective barrier against moisture loss. Looking at the structure of the skin, it is composed of three layers: epidermis, dermis, and subcutaneous fat, and the epidermis is the thinnest of the three layers, and plays an important role in moisturizing and protecting the skin. In particular, the skin becomes elastic and soft by the moisture present in the stratum corneum of the epidermis, and it is known that moisture content of 10% or more is essential to maintain the elasticity of the stratum corneum.
최근 들어, 환경오염 등의 이유로 강한 자외선에 노출되는 경우가 많아졌고, 이에 의한 피부의 수분 손실이 증가하면서 피부 노화가 가속화되고, 피부의 손상에 의한 관련 질병의 발병률이 증가함에 따라, 자외선에 노출된 피부의 보습 유지 및 주름개선 효과를 오랫동안 지속시켜주는 조성물에 대한 요구는 점점 증가하는 추세이다. In recent years, exposure to strong ultraviolet rays has increased due to environmental pollution, and the skin aging is accelerated as the moisture loss of the skin increases, and the incidence of related diseases due to damage to the skin increases. The demand for a composition that maintains the moisturizing and wrinkle-improving effect of the damaged skin for a long time is gradually increasing.
한편, 가오리과에 속하는 홍어는 연골, 저서성 어류로서 우리나라의 흑산도 근해와 서해안에 분포하여 목포와 영광에서 어획되고 있으나, 최근 감소추세에 있어 칠레, 우루과이, 아르헨티나 등지에서도 수입하고 있는 실정이다. 주로 전라남도에서는 홍어를 발효시켜 전통식품으로 애용하고 있으며, 그 독특한 향과 맛을 즐겨 섭취하는 인구가 증가하고 있다. 발효한 홍어를 막걸리와 곁들여 먹는 홍탁이 가장 유명하며, 전남 서남해안 지방에서는 잔치 음식에 삭힌 홍어가 거의 빠지지 않는다.On the other hand, skates belonging to the Stingray family are cartilage, benthic fish, and are distributed in the coasts of Heuksan Island and the west coast of Korea, and are caught in Mokpo and Yeongwang, but are being imported from Chile, Uruguay, and Argentina due to the recent decline. Mainly in Jeollanam-do, fermented skates are used habitually as a traditional food, and the population who enjoys its unique aroma and taste is increasing. Hongtak, which is eaten with fermented skateboard with makgeolli, is the most famous, and in the southwestern coast of Jeollanam-do, fermented skateboard is hardly missed for feast food.
홍어는 바다 심해에서 삼투압 조절을 위해 체내에 질소화합물인 요소 및 트리메틸아민 산화물(trimethylamine oxide)을 많이 함유하고 있는데, 발효에 의해서 이들 성분은 암모니아와 트리메틸아민으로 전환되어 코를 자극하는 강한 향과 톡 쏘는 맛이 생성된다. 홍어는 단백질 19%, 지방 0.9% 정도로 고단백 저지방 어류에 해당하며 상어나 가오리와 같이 베타인(betaine)과 타우린(taurine)이 많이 함유되어 있고, 칼슘과 요소 함량도 높다. 보고된 홍어의 기능적 효과로는 세포막 안정화 작용, 콜레스테롤 조절 작용으로 인한 혈관질환 및 심부전증 예방효과, 성장 발달에 중요한 타우린, 혈청 콜레스테롤 감소와 두뇌성장 발달과 인지기능 향상에 도움을 주는 리놀렌산, 아라키돈산 등 필수 지방산이 함유되어 있으며, EPA와 DHA가 다량 함유되어 관상동맥 질환예방, 혈전증 예방, 두뇌 발달, 시각기능 강화 효과가 있다. 홍어 껍질에 존재하는 콜라겐과 연골에는 뮤코다당 단백질인 콘드로이틴이 함유되어 있어 건강 및 강장 식품으로 알려져 있다. 최근 홍어의 생리활성이나 성분 분석에 대한 연구는 이루어져 있으나, 홍어김치에 존재하는 미생물에 관한 연구는 많이 부족한 실정이다. 홍어김치는 한국 전남지역의 전통 발효식품으로서 기존 배추김치와 차별되는 발효미생물도 다양할 것이다.Skatefish contains a lot of nitrogen compounds, urea and trimethylamine oxide, in the body to control the osmotic pressure in the deep sea, and these components are converted into ammonia and trimethylamine by fermentation, which irritates the nose. A stinging taste is produced. Skate is a high-protein, low-fat fish with 19% protein and 0.9% fat. Like sharks and rays, it contains a lot of betaine and taurine, and has a high content of calcium and urea. The reported functional effects of skate include cell membrane stabilization, cholesterol-regulating effects to prevent vascular disease and heart failure, taurine, which is important for growth and development, linolenic acid and arachidonic acid, which help reduce serum cholesterol and improve brain growth and cognitive function. It contains essential fatty acids, and contains a large amount of EPA and DHA, so it is effective in preventing coronary artery disease, thrombosis, brain development, and visual function. Collagen and cartilage present in the skin of skates contain chondroitin, a mucopolysaccharide protein, and are known as healthy and tonic foods. Recently, studies on the physiological activity and component analysis of skateboards have been conducted, but studies on microorganisms present in skateboard kimchi are insufficient. Hongeo kimchi is a traditional fermented food in the Jeonnam region of Korea, and there will be a variety of fermented microorganisms that are different from existing cabbage kimchi.
한편, 한국등록특허 제1617141호에 항비만 활성을 갖는 페디오코쿠스 이노피나투스가 개시되어 있고, 한국등록특허 제1673450호에 콜레스테롤 저하 효과를 가진 Pediococcus pentosaceus KID7 균주 포함 생균제 조성물이 개시되어 있으나, 본 발명의 페디오코쿠스 이노피나투스를 유효성분으로 포함하는 피부 주름 개선 및 피부 보습용 조성물에 대해 개시된 바 없다.On the other hand, Korean Patent No. 1617141 discloses Pediococcus inofinatus having anti-obesity activity, and Korean Patent No. 1673450 discloses a probiotic composition containing Pediococcus pentosaceus KID7 strain having a cholesterol-lowering effect. There is no disclosure of a composition for improving skin wrinkles and moisturizing skin comprising Pediococcus inofinatus of the present invention as an active ingredient.
본 발명은 상기와 같은 요구에 의해 도출된 것으로서, 본 발명은 페디오코쿠스 이노피나투스를 유효성분으로 포함하는 피부 주름 개선 및 피부 보습용 조성물을 제공하고, 상기 페디오코쿠스 이노피나투스가 자외선 조사에 의해 증진된 MMP-1 및 MMP-9의 발현을 현저하게 억제하고, 피부 각질층을 완화하여 피부두께를 감소시키며, 피부 수분량을 증진시키고, 경피수분 손실량을 감소시키며, 히알루론산의 함량을 증진시킨다는 것을 확인함으로써, 본 발명을 완성하였다.The present invention is derived from the above requirements, the present invention provides a composition for skin wrinkle improvement and skin moisturizing comprising Pediococcus inofinatus as an active ingredient, wherein the Pediococcus inofinatus is irradiated with ultraviolet rays It remarkably inhibits the expression of MMP-1 and MMP-9 promoted by, relieves the stratum corneum of the skin, reduces skin thickness, improves skin moisture, reduces transdermal moisture loss, and increases the content of hyaluronic acid. By confirming this, the present invention was completed.
상기 목적을 달성하기 위하여, 본 발명은 페디오코쿠스 이노피나투스(Pediococcus inopinatus) WiKim0108 균주(기탁번호: KACC 81091BP), 상기 균주의 배양물, 상기 배양물의 농축액, 상기 배양물의 건조물 및 상기 배양물의 추출물 중에서 선택된 1종 이상을 유효성분으로 포함하는 피부 주름 개선 또는 피부 보습용 화장료 조성물을 제공한다.In order to achieve the above object, the present invention is Pediococcus inofinatus ( Pediococcus inopinatus) WiKim0108 strain (Accession Number: KACC 81091BP), for wrinkle or skin moisturizer containing at least one selected from the culture, the culture of water concentrate, the culture of water dried product and extracts the incubation water in the strain as an effective ingredient It provides a cosmetic composition.
또한, 본 발명은 페디오코쿠스 이노피나투스(Pediococcus inopinatus) WiKim0108 균주, 상기 균주의 배양물, 상기 배양물의 농축액, 상기 배양물의 건조물 및 상기 배양물의 추출물 중에서 선택된 1종 이상을 유효성분으로 포함하는 피부 주름 개선 또는 피부 보습용 건강기능식품 조성물을 제공한다.In addition, the present invention Pediococcus inofinatus ( Pediococcus provides inopinatus) WiKim0108 strain, wrinkle or skin-friendly functional food composition comprising at least one member selected from the culture, the culture of water concentrate, the culture of water dried product and extracts the incubation water in the strain as an effective ingredient .
또한, 본 발명은 페디오코쿠스 이노피나투스(Pediococcus inopinatus) WiKim0108 균주, 상기 균주의 배양물, 상기 배양물의 농축액, 상기 배양물의 건조물 및 상기 배양물의 추출물 중에서 선택된 1종 이상을 유효성분으로 포함하는 피부 건조증의 예방 또는 치료용 약학 조성물을 제공한다.In addition, the present invention Pediococcus inofinatus ( Pediococcus It provides inopinatus) WiKim0108 strain, a pharmaceutical composition for the prevention or treatment of dry skin, including at least one selected from the culture, wherein the culture concentrate water, the culture of water and extract the dried product of the water culture strain as an effective ingredient.
또한, 본 발명은 페디오코쿠스 이노피나투스(Pediococcus inopinatus) WiKim0108 균주(기탁번호: KACC 81091BP), 상기 균주의 배양물, 상기 배양물의 농축액, 상기 배양물의 건조물 및 상기 배양물의 추출물 중에서 선택된 1종 이상을 유효성분으로 포함하는 피부 주름 개선 또는 피부 보습용 사료 조성물을 제공한다.In addition, the present invention Pediococcus inofinatus ( Pediococcus inopinatus) WiKim0108 strain (Accession Number: KACC 81091BP), for wrinkle or skin moisturizer containing at least one selected from the culture, the culture of water concentrate, the culture of water dried product and extracts the incubation water in the strain as an effective ingredient Provide a feed composition.
또한, 본 발명은 페디오코쿠스 이노피나투스(Pediococcus inopinatus) WiKim0108 균주(기탁번호: KACC 81091BP), 상기 균주의 배양물, 상기 배양물의 농축액, 상기 배양물의 건조물 및 상기 배양물의 추출물 중에서 선택된 1종 이상을 유효성분으로 포함하는 피부 주름 개선 또는 피부 보습용 수의학적 조성물을 제공한다.In addition, the present invention Pediococcus inofinatus ( Pediococcus inopinatus) WiKim0108 strain (Accession Number: KACC 81091BP), for wrinkle or skin moisturizer containing at least one selected from the culture, the culture of water concentrate, the culture of water dried product and extracts the incubation water in the strain as an effective ingredient Veterinary compositions are provided.
본 발명은 페디오코쿠스 이노피나투스를 유효성분으로 포함하는 피부 주름 개선 및 피부 보습용 조성물에 관한 것으로, 상세하게는 상기 페디오코쿠스 이노피나투스는 자외선 조사에 의해 증진된 MMP-1 및 MMP-9의 발현을 현저하게 억제하고, 피부 각질층을 완화하여 피부두께를 감소시키며, 피부 수분량을 증진시키고, 경피수분 손실량을 감소시키며, 히알루론산의 함량을 증진시키는 효과가 있다.The present invention relates to a composition for improving skin wrinkles and moisturizing skin comprising Pediococcus inofinatus as an active ingredient, and in detail, the Pediococcus inofinatus is MMP-1 and MMP- enhanced by UV irradiation. It has the effect of remarkably suppressing the expression of 9, reducing the skin thickness by alleviating the stratum corneum of the skin, improving the amount of skin moisture, reducing the amount of transdermal moisture loss, and increasing the content of hyaluronic acid.
도 1은 본 발명의 페디오코쿠스 이노피나투스 처리에 따른 MMP-1 및 MMP-9의 발현량 변화를 확인한 결과이다. ####은 정상군 대비 자외선 조사군의 MMP-1 및 MMP-9의 발현량이 통계적으로 유의미하게 증가하였다는 것으로, p<0.0001이고, ****은 자외선 조사군에 비해 본 발명의 페디오코쿠스 이노피나투스 처리군의 MMP-1 및 MMP-9의 발현량이 통계적으로 유의미하게 감소하였다는 것으로, p<0.0001이다.
도 2는 본 발명의 페디오코쿠스 이노피나투스(WiKim0108)를 투여한 무모 생쥐군으로부터 채취한 조직의 염색결과를 나타낸 것이다. (A)는 H&E 염색결과이고, (B)는 Masson's trichrome 염색결과이며, (C)는 H&E 염색을 실시한 조직의 케라틴층에서 표피 기저막까지의 두께(Epidermis thickness)를 나타낸 결과이다. ####은 정상군(Nor) 대비 자외선 조사군(Veh)의 표피 두께가 통계적으로 유의하게 증가하였다는 것으로, p<0.0001이고, ***는 자외선 조사군(Veh) 대비 본 발명의 페디오코쿠스 이노피나투스 투여군이 통계적으로 유의미하게 표피 두께가 감소하였다는 것으로 p<0.001이다.
도 3은 본 발명의 페디오코쿠스 이노피나투스(WiKim0108)의 투여에 의한 경피 수분손실량(TEWL) 억제효능을 나타낸 그래프이다. ####는 정상군(Nor) 대비 자외선 조사군(Veh)의 경피 수분손실량이 통계적으로 유의미하게 증가하였다는 것으로, p<0.0001이고, ****는 자외선 조사군(Veh) 대비 본 발명의 페디오코쿠스 이노피나투스 투여군(WiKim0108)의 경피 수분손실량이 통계적으로 유의미하게 감소하였다는 것으로, p<0.0001이다.
도 4는 본 발명의 페디오코쿠스 이노피나투스(WiKim0108)의 투여에 의한 피부 수분량(Capacitance)을 나타낸 그래프이다. ####는 정상군(Nor) 대비 자외선 조사군(Veh)의 피부 수분량이 통계적으로 유의미하게 감소하였다는 것으로, p<0.0001이고, ***는 자외선 조사군(Veh) 대비 본 발명의 페디오코쿠스 이노피나투스 투여군(WiKim0108)의 피부 수분량이 통계적으로 유의미하게 증가하였다는 것으로, p<0.001이다.
도 5는 본 발명의 페디오코쿠스 이노피나투스를 처리한 마우스 피부조직에서의 히알루론산 함량을 분석한 결과이다. ####는 정상군(Nor) 대비 자외선 조사군(Veh)의 히알루론산의 함량이 통계적으로 유의미하게 감소하였다는 것으로, p<0.0001이고, ****는 자외선 조사군(Veh) 대비 본 발명의 페디오코쿠스 이노피나투스 투여군(WiKim0108)의 히알루론산의 함량이 통계적으로 유의미하게 증가하였다는 것으로, p<0.0001이다.1 is a result of confirming the change in the expression levels of MMP-1 and MMP-9 according to the treatment of Pediococcus inofinatus of the present invention. #### indicates that the expression levels of MMP-1 and MMP-9 in the UV-irradiated group were statistically significantly increased compared to the normal group, p<0.0001, and **** is the pedi of the present invention compared to the UV-irradiated group The expression levels of MMP-1 and MMP-9 in the Okoccus inofinatus treatment group were statistically significantly decreased, and p<0.0001.
Figure 2 shows the staining results of tissues collected from hairless mice group administered with Pediococcus inofinatus (WiKim0108) of the present invention. (A) is the result of H&E staining, (B) is the result of Masson's trichrome staining, and (C) is the result showing the epidermis thickness from the keratin layer of the tissue subjected to H&E staining to the epidermal basement membrane. #### indicates that the epidermal thickness of the ultraviolet irradiation group (Veh) was statistically significantly increased compared to the normal group (Nor), p<0.0001, and *** is the pedi of the present invention compared to the ultraviolet irradiation group (Veh). It is p<0.001 that the epidermal thickness of the Ococus inofinatus administration group decreased statistically significantly.
3 is a graph showing the effect of inhibiting transdermal water loss (TEWL) by administration of Pediococcus inofinatus (WiKim0108) of the present invention. #### indicates that the transdermal water loss of the ultraviolet irradiation group (Veh) was statistically significantly increased compared to the normal group (Nor), p<0.0001, and **** is the present invention compared to the ultraviolet irradiation group (Veh) The amount of percutaneous water loss of the Pediococcus inofinatus administration group (WiKim0108) was statistically significantly reduced, and p<0.0001.
Figure 4 is a graph showing the skin moisture content (Capacitance) by administration of Pediococcus inofinatus (WiKim0108) of the present invention. #### indicates that the skin moisture content of the ultraviolet irradiation group (Veh) was statistically significantly decreased compared to the normal group (Nor), p<0.0001, and *** is the pedi of the present invention compared to the ultraviolet irradiation group (Veh). It was found that the skin moisture content of the Okoccus inofinatus administration group (WiKim0108) increased statistically significantly, and p<0.001.
5 is a result of analyzing the hyaluronic acid content in the skin tissue of the mouse treated with Pediococcus inofinatus of the present invention. #### indicates that the content of hyaluronic acid in the ultraviolet irradiation group (Veh) was statistically significantly reduced compared to the normal group (Nor), p<0.0001, and **** is the original compared to the ultraviolet irradiation group (Veh). The content of hyaluronic acid in the group administered with Pediococcus inofinatus (WiKim0108) of the present invention was statistically significant, indicating that p<0.0001.
본 발명은 페디오코쿠스 이노피나투스(Pediococcus inopinatus) WiKim0108 균주 (기탁번호: KACC 81091BP), 상기 균주의 배양물, 상기 배양물의 농축액, 상기 배양물의 건조물 및 상기 배양물의 추출물 중에서 선택된 1종 이상을 유효성분으로 포함하는 피부 주름 개선 또는 피부 보습용 화장료 조성물에 관한 것이다.The present invention is Pediococcus inofinatus ( Pediococcus inopinatus) WiKim0108 strain (Accession Number: KACC 81091BP), for wrinkle or skin moisturizer containing at least one selected from the culture, the culture of water concentrate, the culture of water dried product and extracts the incubation water in the strain as an effective ingredient It relates to a cosmetic composition.
상기 페디오코쿠스 이노피나투스 WiKim0108 균주는 홍어 또는 홍어김치로부터 분리된 것이 바람직하지만, 이에 한정하는 것은 아니다. The Pediococcus Inofinatus WiKim0108 strain is preferably isolated from skateboard or skate kimchi, but is not limited thereto.
상기 유효성분은 MMP-1 및 MMP-9 단백질의 발현량을 감소시키며, 히알루론산의 함량을 증가시키는 것이 특징이다. The active ingredient is characterized by reducing the expression levels of MMP-1 and MMP-9 proteins, and increasing the content of hyaluronic acid.
본 발명의 일 구현 예에 따른 화장료 조성물은 크림, 유연화장수, 영양화장수, 팩, 에센스, 헤어토닉, 샴푸, 린스, 헤어 컨디셔너, 헤어 트리트먼트, 젤, 스킨로션, 스킨소프너, 스킨토너, 아스트린젠트, 밀크로션, 모이스처 로션, 영양로션, 마사지 크림, 영양크림, 모이스처 크림, 핸드 크림, 파운데이션, 영양에센스, 선스크린, 비누, 클렌징폼, 클렌징로션, 클렌징크림, 바디 로션 및 바디 클렌저로 이루어지는 군으로부터 선택된 어느 하나의 제형을 가질 수 있으나, 이에 제한되지 않는다. 이들 각 제형으로 이루어진 화장료 조성물은 그 제형의 제제화에 필요하고 적절한 각종의 기제와 첨가물을 함유할 수 있으며, 이들 성분의 종류와 양은 당업자에 의해 용이하게 선정될 수 있다.A cosmetic composition according to an embodiment of the present invention is a cream, softening lotion, nutrient lotion, pack, essence, hair tonic, shampoo, conditioner, hair conditioner, hair treatment, gel, skin lotion, skin softener, skin toner, astringent, Selected from the group consisting of milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, foundation, nutrition essence, sunscreen, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion and body cleanser. It may have any one formulation, but is not limited thereto. The cosmetic composition composed of each of these formulations may contain various bases and additives necessary and appropriate for the formulation of the formulation, and the types and amounts of these components can be easily selected by those skilled in the art.
본 발명의 화장료 조성물의 제형이 페이스트, 크림 또는 겔인 경우에는 담체 성분으로서 동물섬유, 식물섬유, 왁스, 파라핀, 전분, 트라가칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산화아연 등이 이용될 수 있다.When the formulation of the cosmetic composition of the present invention is a paste, cream or gel, animal fibers, plant fibers, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide as carrier components Etc. may be used.
본 발명의 화장료 조성물의 제형이 파우더 또는 스프레이인 경우에는 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄 히드록시드, 칼슘 실리케이트 또는 폴리아미드 파우더가 이용될 수 있고, 특히 스프레이인 경우에는 추가로 클로로플루오로히드로카본, 프로판-부탄 또는 디메틸 에테르와 같은 추진체를 포함할 수 있다.When the formulation of the cosmetic composition of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In particular, in the case of a spray, additional chlorofluoro Propellants such as hydrocarbon, propane-butane or dimethyl ether.
본 발명의 화장료 조성물의 제형이 용액 또는 유탁액의 경우에는 담체 성분으로서 용매, 용매화제 또는 유탁화제가 이용되고, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌글리콜, 1, 3-부틸글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜 또는 소르비탄의 지방산 에스테르가 있다.When the formulation of the cosmetic composition of the present invention is a solution or emulsion, a solvent, a solvating agent or an emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene Glycol, 1,3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan.
본 발명의 화장료 조성물의 제형이 현탁액인 경우에는 담체 성분으로서 물, 에탄올 또는 프로필렌 글리콜과 같은 액상 희석제, 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르 및 폴리옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미소결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가 또는 트라칸트 등이 이용될 수 있다.When the formulation of the cosmetic composition of the present invention is 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 may be used.
본 발명의 화장료 조성물의 제형이 계면-활성제 함유 클렌징인 경우에는 담체 성분으로서 지방족 알코올 설페이트, 지방족 알코올 에테르설페이트, 설포숙신산 모노에스테르, 아세티오네이트, 이미다졸리늄 유도체, 메틸타우레이트, 사르코시네이트, 지방산 아미드 에테르 설페이트, 알킬아미도베타인, 지방족 알코올, 지방산 글리세리드, 지방산 디에탄올아미드, 식물성 유, 리놀린 유도체 또는 에톡실화 글리세롤 지방산 에스테르 등이 이용될 수 있다.When the formulation of the cosmetic composition of the present invention is a surfactant containing cleansing, as carrier components, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, acetionate, imidazolinium derivative, methyltaurate, sarcosinate , Fatty acid amide ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty diethanolamide, vegetable oil, linoline derivative or ethoxylated glycerol fatty acid ester, and the like may be used.
본 발명의 화장료 조성물은 형광물질, 살진균제, 굴수성 유발물질, 보습제, 방향제, 방향제 담체, 단백질, 용해화제, 당 유도체, 일광차단제, 비타민, 식물 추출물 등을 포함하는 부형제를 추가로 함유할 수 있다.The cosmetic composition of the present invention may further contain excipients including fluorescent substances, fungicides, hydrophobic substances, moisturizers, fragrances, fragrance carriers, proteins, solubilizers, sugar derivatives, sunscreen agents, vitamins, plant extracts, etc. .
또한, 본 발명은 페디오코쿠스 이노피나투스(Pediococcus inopinatus) WiKim0108 균주, 상기 균주의 배양물, 상기 배양물의 농축액, 상기 배양물의 건조물 및 상기 배양물의 추출물 중에서 선택된 1종 이상을 유효성분으로 포함하는 피부 주름 개선 또는 피부 보습용 건강기능식품 조성물에 관한 것이다.In addition, the present invention Pediococcus inofinatus ( Pediococcus relates to inopinatus) WiKim0108 strain, the culture, the culture of water concentrate of the strain, wrinkle or skin moisturizing dietary supplement composition comprising the culture of water dried form and the cultured at least one selected from water extract as an active ingredient .
상기 유효성분은 분말, 과립, 환, 정제, 캡슐, 캔디, 시럽 및 음료 중에서 선택된 어느 하나의 제형으로 제조되는 것이 바람직하지만 이에 한정하지 않는다.The active ingredient is preferably prepared in any one formulation selected from powder, granule, pill, tablet, capsule, candy, syrup, and beverage, but is not limited thereto.
상기에서 언급한 본 발명의 균주 또는 이의 배양액 등을 건강기능식품 조성물로 사용할 경우, 상기 균주 또는 이의 배양액을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용할 수 있으며, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효 성분의 혼합양은 그 사용 목적 (예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다.When using the above-mentioned strain of the present invention or a culture solution thereof as a health functional food composition, the strain or a culture solution thereof may be added as it is, or may be used with other foods or food ingredients, and may be appropriately used according to a conventional method. have. The mixing amount of the active ingredient may be appropriately determined according to the purpose of use (prevention, health or therapeutic treatment).
본 발명의 건강기능식품 조성물은 식품 제조 시에 통상적으로 첨가되고 식품학적으로 허용되는 성분을 더 포함할 수 있다. 예컨대, 음료수로 제조되는 경우에는 본 발명의 균주 이외에 구연산, 액상과당, 설탕, 포도당, 초산, 사과산, 과즙 등에서 하나 이상의 성분을 추가로 포함할 수 있다.The health functional food composition of the present invention may further include ingredients that are commonly added during food production and food wise acceptable. For example, when prepared as a beverage, it may further include one or more components from citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, etc. in addition to the strain of the present invention.
본 발명에 따른 건강기능식품 조성물의 유효성분으로 포함될 수 있는 양은 피부 주름 개선 또는 피부 보습용 건강기능식품을 원하는 사람의 연령, 성별, 체중, 상태, 질병의 증상에 따라 적절히 선택될 수 있으며, 바람직하게는 성인기준 1일 0.01 내지 10.0g 정도로 포함되는 것이 좋으며, 이러한 함량을 갖는 식품을 섭취함으로써 피부 주름 개선 또는 피부 보습 증진 효과를 얻을 수 있다.The amount that can be included as an active ingredient of the health functional food composition according to the present invention may be appropriately selected according to the age, sex, weight, condition, and symptoms of a disease of a person who wants a health functional food for skin wrinkle improvement or skin moisturization, and is preferable It is recommended to contain about 0.01 to 10.0g per day for adults, and by ingesting foods having such a content, it is possible to improve skin wrinkles or improve skin moisturization.
또한, 본 발명은 페디오코쿠스 이노피나투스(Pediococcus inopinatus) WiKim0108 균주, 상기 균주의 배양물, 상기 배양물의 농축액, 상기 배양물의 건조물 및 상기 배양물의 추출물 중에서 선택된 1종 이상을 유효성분으로 포함하는 피부 건조증의 예방 또는 치료용 약학 조성물에 관한 것이다.In addition, the present invention Pediococcus inofinatus ( Pediococcus inopinatus) WiKim0108 strain, the present invention relates to at least one selected from the culture, wherein the culture concentrate water, the culture of water and extract the dried product of the water culture strain to a pharmaceutical composition for the prevention or treatment of dry skin which comprises as an active ingredient.
본 발명에서의 용어 '예방'이란 본 발명에 따른 약학적 조성물의 투여에 의해 피부 건조증의 발병을 억제 또는 지연시키는 모든 행위를 의미하고, '치료'란 상기 약학적 조성물의 투여에 의해 피부 건조증의 의심 및 발병 개체의 증상이 호전되거나 이롭게 변경되는 모든 행위를 의미한다. 본 발명의 상기 약학조성물은 약학적 조성물의 제조에 통상적으로 사용하는 적절한 담체, 부형제 및 희석제를 더 포함할 수 있다. 본 발명에 따른 상기 약학조성물은, 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화하여 사용될 수 있다. 본 발명에 따른 상기 약학조성물에 포함될 수 있는 담체, 부형제 및 희석제로는 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유 등을 포함한 다양한 화합물 혹은 혼합물을 들 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구투여를 위한 고형 제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형 제제는 상기 균주 또는 상기 균주 유래 소포체에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘카보네이트(calcium carbonate), 수크로스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제된다. 또한, 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순 희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제가 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜(propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로젤라틴 등이 사용될 수 있다.The term'prevention' in the present invention refers to any action that suppresses or delays the onset of dry skin by administration of the pharmaceutical composition according to the present invention, and'treatment' refers to the treatment of dry skin by administration of the pharmaceutical composition. It refers to any action in which symptoms of suspected and affected individuals are improved or beneficially altered. The pharmaceutical composition of the present invention may further include suitable carriers, excipients, and diluents commonly used in the preparation of pharmaceutical compositions. The pharmaceutical compositions according to the present invention are formulated in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, and sterile injectable solutions according to a conventional method. Can be used. Carriers, excipients and diluents that can be included in the pharmaceutical composition according to the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, Various compounds or mixtures including calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. . In the case of formulation, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants that are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient, such as starch, calcium carbonate, in the strain or the endoplasmic reticulum derived from the strain. , Sucrose (sucrose) or lactose (lactose), gelatin, etc. are mixed and prepared. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, liquid solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweetening agents, fragrances, and preservatives may be included. . Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerogelatin, and the like may be used.
본 발명의 약학 조성물은 약제학적으로 유효한 양으로 투여될 수 있는데, 본 발명의 용어 '약제학적으로 유효한 양'이란 의학적 예방 또는 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 예방 또는 치료하기에 충분한 양을 의미하며, 유효 용량 수준은 질환의 중증도, 약물의 활성, 환자의 연령, 체중, 건강, 성별, 환자의 약물에 대한 민감도, 사용된 본 발명 조성물의 투여 시간, 투여 경로 및 배출 비율, 치료기간, 사용된 본 발명의 조성물과 배합 또는 동시 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 결정될 수 있다. 본 발명의 약학적 조성물은 개별 치료제로 투여하거나 다른 치료제와 병용하여 투여될 수 있고 종래의 치료제와는 순차적으로 또는 동시에 투여될 수 있다. 그리고 단일 또는 다중 투여될 수 있다. 상기 요소를 모두 고려하여 부작용없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하다.The pharmaceutical composition of the present invention may be administered in a pharmaceutically effective amount, and the term'pharmaceutically effective amount' of the present invention is sufficient to prevent or treat a disease at a reasonable benefit/risk ratio applicable to medical prevention or treatment. It means an amount, and the effective dose level is the severity of the disease, the activity of the drug, the patient's age, weight, health, sex, the patient's sensitivity to the drug, the time of administration of the composition of the present invention used, the route of administration and the rate of excretion, treatment The duration, factors including drugs used in combination or co-use with the composition of the present invention used, and other factors well known in the medical field. The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or administered in combination with other therapeutic agents, and may be administered sequentially or simultaneously with a conventional therapeutic agent. And can be administered single or multiple. Considering all of the above factors, it is important to administer an amount capable of obtaining the maximum effect in a minimum amount without side effects.
본 발명의 조성물의 투여 빈도는 특별히 이에 제한되지 않으나, 1일 1회 투여하거나 또는 용량을 분할하여 수회 투여할 수 있다. 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다.The frequency of administration of the composition of the present invention is not particularly limited thereto, but may be administered once a day or divided into several doses. The above dosage does not in any way limit the scope of the present invention.
본 발명의 용어 '개체'란 피부 건조증이 발병될 가능성이 있거나 또는 발병된 쥐, 가축, 인간 등을 포함하는 포유동물을 제한 없이 포함한다.The term “individual” of the present invention includes, without limitation, mammals, including mice, livestock, humans, etc. that may develop skin dryness or are affected.
본 발명의 피부 건조증의 예방 또는 치료 방법에 있어서, 상기 약학적 조성물의 투여 경로는 목적 조직에 도달할 수 있는 한 어떠한 일반적인 경로를 통하여도 투여될 수 있다. 본 발명의 약학적 조성물은 특별히 이에 제한되지 않으나, 경구 투여, 직장 내 투여 등의 경로를 통해 투여할 수 있으며 경우에 따라 목적에 따라 다른 경로로도 투여될 수 있다. 다만, 경구 투여 시에는 위산에 의하여 상기 WiKim0108 균주가 변성될 수 있기 때문에 경구용 조성물은 활성 약제를 코팅하거나 위에서의 분해로부터 보호되도록 제형화 되어야 한다. 또한, 상기 조성물은 활성 물질이 표적 세포로 이동할 수 있는 임의의 장치에 의해 투여될 수 있다.In the method for preventing or treating dry skin of the present invention, the route of administration of the pharmaceutical composition may be administered through any general route as long as it can reach the target tissue. The pharmaceutical composition of the present invention is not particularly limited thereto, but may be administered through a route such as oral administration or rectal administration, and in some cases, may be administered by other routes depending on the purpose. However, since the WiKim0108 strain may be denatured by gastric acid when administered orally, the oral composition should be coated with an active agent or formulated to protect it from degradation in the stomach. In addition, the composition can be administered by any device capable of moving the active substance to the target cell.
또한, 본 발명은 페디오코쿠스 이노피나투스(Pediococcus inopinatus) WiKim0108 균주(기탁번호: KACC 81091BP), 상기 균주의 배양물, 상기 배양물의 농축액, 상기 배양물의 건조물 및 상기 배양물의 추출물 중에서 선택된 1종 이상을 유효성분으로 포함하는 피부 주름 개선 또는 피부 보습용 사료 조성물에 관한 것이다.In addition, the present invention Pediococcus inopinatus ( Pediococcus inopinatus) WiKim0108 strain (accession number: KACC 81091BP), at least one selected from the culture of the strain, the concentrate of the culture, a dried product of the culture and an extract of the culture It relates to a feed composition for skin wrinkle improvement or skin moisturizing, comprising as an active ingredient.
상기 균주에 대해서는 전술한 바와 같으며, 피부 주름 개선 또는 피부 보습을 목적으로 사료 조성물로 첨가할 수 있다. 상기 사료용 조성물은 사료 첨가제를 포함할 수 있다. 본 발명의 사료첨가제는 사료관리법상의 보조사료에 해당한다.The strain is as described above, and may be added as a feed composition for the purpose of improving skin wrinkles or moisturizing the skin. The feed composition may contain a feed additive. The feed additive of the present invention corresponds to an auxiliary feed in the feed management law.
본 발명에서 용어 '사료'는 동물이 먹고, 섭취하며, 소화시키기 위한 또는 이에 적당한 임의의 천연 또는 인공 규정식, 한끼식 등 또는 상기 한끼식의 성분을 의미할 수 있다. 상기 사료의 종류는 특별히 제한되지 아니하며, 당해 기술 분야에서 통상적으로 사용되는 사료를 사용할 수 있다. 상기 사료의 비제한적인 예로는, 곡물류, 근과류, 식품 가공 부산물류, 조류, 섬유질류, 제약 부산물류, 유지류, 전분류, 박류 또는 곡물 부산물류 등과 같은 식물성 사료; 단백질류, 무기물류, 유지류, 광물성류, 유지류, 단세포 단백질류, 동물성 플랑크톤류 또는 음식물 등과 같은 동물성 사료를 들 수 있다. 이들은 단독으로 사용되거나 2종 이상을 혼합하여 사용될 수 있다.In the present invention, the term'feed' may mean any natural or artificial diet, one meal meal, etc., or a component of the one meal meal for animals to eat, ingest, and digest. The kind of feed is not particularly limited, and feed commonly used in the art may be used. Non-limiting examples of the feed include vegetable feeds such as grains, root fruits, food processing by-products, algae, fiber, pharmaceutical by-products, oils and fats, starches, meals or grain by-products; Animal feeds such as proteins, inorganic logistics, oils and fats, minerals, oils and fats, single-cell proteins, zooplanktons or foods. These may be used alone or in combination of two or more.
또한, 본 발명은 페디오코쿠스 이노피나투스(Pediococcus inopinatus) WiKim0108 균주(기탁번호: KACC 81091BP), 상기 균주의 배양물, 상기 배양물의 농축액, 상기 배양물의 건조물 및 상기 배양물의 추출물 중에서 선택된 1종 이상을 유효성분으로 포함하는 피부 주름 개선 또는 피부 보습용 수의학적 조성물에 관한 것이다.In addition, the present invention Pediococcus inofinatus ( Pediococcus inopinatus) WiKim0108 strain (Accession Number: KACC 81091BP), for wrinkle or skin moisturizer containing at least one selected from the culture, the culture of water concentrate, the culture of water dried product and extracts the incubation water in the strain as an effective ingredient It relates to a veterinary composition.
본 발명의 수의학적 조성물에 포함될 수 있는 부형제 및 희석제로는, 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다. 본 발명의 수의학적 조성물은 충진제, 항응집제, 윤활제, 습윤제, 향신료, 유화제, 방부제 등을 추가로 포함할 수 있는데, 본 발명에 의한 수의학적 조성물이 동물에 투여된 후 활성 성분의 신속, 지속 또는 지연된 방출을 제공할 수 있도록 당업계에 잘 알려진 방법을 사용하여 제형화될 수 있고, 제형은 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 용액, 시럽, 에어로졸, 연질 또는 경질 젤라틴 캅셀, 좌제, 멸균 주사용액, 멸균 외용제 등의 형태일 수 있다. 본 발명에 의한 수의학적 조성물은 동물의 나이, 성별, 체중에 따라 달라질 수 있으며, 1일 1회 내지 수회 투여할 수 있고, 투여량은 투여경로, 성별, 체중, 나이 등에 따라서 증감될 수 있다. 따라서 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다.Excipients and diluents that may be included in the veterinary composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate. , Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oils. The veterinary composition of the present invention may further include a filler, an anti-aggregating agent, a lubricant, a wetting agent, a spice, an emulsifying agent, a preservative, and the like. After the veterinary composition according to the present invention is administered to an animal, the active ingredient is rapidly It can be formulated using methods well known in the art to provide delayed release, and the formulations are powders, granules, tablets, capsules, suspensions, emulsions, solutions, syrups, aerosols, soft or hard gelatin capsules, suppositories. , Sterile injectable solutions, sterile external preparations, and the like. The veterinary composition according to the present invention may vary depending on the age, sex, and weight of the animal, and may be administered once to several times a day, and the dosage may be increased or decreased depending on the route of administration, sex, weight, age, and the like. Therefore, the dosage does not limit the scope of the present invention in any way.
이하, 실시예를 이용하여 본 발명을 더욱 상세하게 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로 본 발명의 범위가 이들에 의해 제한되지 않는다는 것은 당해 기술분야에서 통상의 지식을 가진 자에게 있어 자명한 것이다. Hereinafter, the present invention will be described in more detail using examples. These examples are only for describing the present invention in more detail, and it is apparent to those of ordinary skill in the art that the scope of the present invention is not limited thereto.
실시예Example 1. One. 페디오코쿠스Pediococcus 이노피나투스Inofinatus (( PediococcusPediococcus inopinatusinopinatus ) ) WiKim0108WiKim0108 의 분리 및 동정Separation and identification of
(1) (One) 신균주Shin Gyunju 분리 Separation
본 발명에서 사용한 신균주는 전라남도 나주시 영산포 홍어 영농조합법인으로부터 구입한 홍어김치(10g)를 취하여 멸균 생리식염수로 십진 희석한 후, MRS 유산균 분리용 고체배지에 도말하여 30℃에서 48시간 동안 배양 후에 집락의 형태에 따라 서로 다른 미생물을 분리하여 사용하였다. 분리한 미생물은 영양 최소배지(1% 글루코오스, 0.5% 효모 추출물)에서 30℃에서 48시간 동안 배양하였다. The new strain used in the present invention takes skate kimchi (10g) purchased from the Skate Farming Association in Youngsanpo, Naju, Jeollanam-do, diluted with sterilized physiological saline, spread on a solid medium for separating MRS lactic acid bacteria, and incubated at 30℃ for 48 hours. Different microorganisms were separated and used according to the type of colony. The isolated microorganisms were cultured at 30° C. for 48 hours in minimal nutrient medium (1% glucose, 0.5% yeast extract).
(2) 유산균의 동정(2) Identification of lactic acid bacteria
홍어김치 미생물 WiKim0108은 16S ribosomal DNA 유전자 서열 분석을 통하여 동정하였다. 선발 미생물의 균체를 취하고 멸균된 생리식염수에 균체를 2회 세척한 후, DNeasy 조직 키트(Qiagen, Valecia, CA, USA)를 사용하여 DNA를 추출하였다. 추출된 DNA의 16S ribosomal DNA 유전자 증폭을 위하여 유니버설 프라이머; 27F (5'-AGAGTTTGATCATGGCTCAG-3')(서열번호 1)와 1492R (5'-GGATACCTTGTTACGACTT-3')(서열번호 2) 프라이머를 사용하였다. PCR 반응시 0.4mM dNTP, 0.5 유닛의 Taq 폴리머라아제, 4mM Mg2 +이 함유된 다카라 Perfect Premix(Takara, Japan) 10㎕에, DNA template(20㎍/㎖) 1㎕, 1.0μM 정방향 프라이머 및 1.0μM 역방향 프라이머를 각각 1㎕씩 넣고 나머지는 증류수를 첨가하여 총 부피가 20㎕가 되도록 제조하였다. The skate kimchi microorganism WiKim0108 was identified through 16S ribosomal DNA gene sequence analysis. After taking the cells of the selected microorganism and washing the cells twice in sterilized physiological saline, DNA was extracted using a DNeasy tissue kit (Qiagen, Valecia, CA, USA). Universal primer for amplification of 16S ribosomal DNA gene of extracted DNA; 27F (5'-AGAGTTTGATCATGGCTCAG-3') (SEQ ID NO: 1) and 1492R (5'-GGATACCTTGTTACGACTT-3') (SEQ ID NO: 2) primers were used. In the PCR reaction, in 10 μl of Takara Perfect Premix (Takara, Japan) containing 0.4mM dNTP, 0.5 unit of Taq polymerase, 4mM Mg 2 + , 1 μl of DNA template (20 μg/ml), 1.0 μM forward primer and 1.0 μM reverse primers were added at 1 μl each, and distilled water was added to the rest to obtain a total volume of 20 μl.
PCR 증폭은 마스터사이클러 그래디언트(Mastercycler gradient)(Eppendorf, Hamburg, Germany)으로 수행하였다. PCR 반응은 95℃에서 5분(initial denaturation), 94℃에서 45초(denaturation), 52℃에서 45초(annealing), 72℃에서 1분(extension)을 30 cycles 실시하였고, 72℃에서 5분 동안 최종 연장(extension)을 실시하였다. 증폭된 약 1400bp의 fragment를 T 벡터(Invitrogen, Carlsbad, CA, USA)에 결합시킨 후 형질전환하였다. T 벡터 시퀀싱 프라이머를 이용하여 염기서열 결정을 수행하였으며, 그 결과는 BLAST 분석 프로그램(http://www.ncbi.nlm.nih.gov)을 이용하여 GenBank (NCBI, Bethesda, MD, USA)의 ribosomal DNA 유전자 시퀀싱과 비교하여 동정하였다. 결정된 염기서열은 Clustal_W 및 BioEdit 프로그램으로 정렬하였고, 발생한 gap은 이후 분석과정에서 결여형질로 처리하였다. 염기서열 간의 유전적 거리 계산과 neighbor-joining, maximum-likelihood 및 parsimony 계통수의 제작은 Mega 프로그램을 사용하여 수행하였고 계통군의 지지도를 알아보기 위해 위의 세 가지 분석에 대해 각각 동일한 사양으로 bootstrap 분석하였다(1,000 replicates). PCR amplification was performed with a Mastercycler gradient (Eppendorf, Hamburg, Germany). The PCR reaction was performed for 30 cycles at 95°C for 5 minutes (initial denaturation), 94°C for 45 seconds (denaturation), 52°C for 45 seconds (annealing), 72°C for 1 minute (extension), and 72°C for 5 minutes. During the final extension (extension) was carried out. The amplified fragment of about 1400bp was transformed after binding to a T vector (Invitrogen, Carlsbad, CA, USA). Sequencing was performed using T vector sequencing primers, and the results were ribosomal of GenBank (NCBI, Bethesda, MD, USA) using BLAST analysis program (http://www.ncbi.nlm.nih.gov). It was identified by comparison with DNA gene sequencing. The determined nucleotide sequence was aligned with Clustal_W and BioEdit programs, and the generated gap was treated as a missing trait in the subsequent analysis process. The calculation of the genetic distance between nucleotide sequences and the generation of neighbor-joining, maximum-likelihood and parsimony phylogenies were performed using the Mega program, and bootstrap analysis was performed with the same specifications for each of the above three analyses to determine the support of the phylogeny. (1,000 replicates).
홍어김치로부터 분리한 유산균 WiKim0108 균주의 동정 및 분류를 위하여, 16S rRNA 염기서열을 분석한 결과, 신균주 WiKim0108은 Pediococcus inopinatus 표준 균주 BT와 97% 일치하였다(서열번호 3). 이후 상기 신균주를 페디오코쿠스 이노피나투스 WiKim0108(Pediococcus inopinatus WiKim0108)로 명명하였다. 또한, 상기 페디오코쿠스 이노피나투스 WiKim0108을 국립농업과학원 농업유전자원센터에 2019년 3월 8일자로 기탁하여, 기탁번호 KACC 81091BP를 부여받았다.For the identification and classification of the lactic acid bacteria WiKim0108 strain isolated from skate kimchi, 16S rRNA nucleotide sequence analysis showed that the new strain WiKim0108 was Pediococcus. Inopinatus was 97% consistent with the standard strain BT (SEQ ID NO: 3). Then, the new strain was added to Pediococcus Inofinatus WiKim0108 ( Pediococcus It was named inopinatus WiKim0108). In addition, the Pediococcus Inofinatus WiKim0108 was deposited with the Agricultural Genetic Resource Center of the National Academy of Agricultural Sciences on March 8, 2019, and was given the deposit number KACC 81091BP.
실시예Example 2. 2. MMPMMP -1 및 -1 and MMPMMP -9 단백질의 발현량 변화 분석-9 Protein expression level change analysis
MMP-1 및 MMP-9는 피부노화와 연관있는 단백질로 자외선과 같은 산화적 스트레스에 의해 증가하는 것으로 알려졌으므로 본 실험에서는 산화적 손상으로부터 유도된 MMP-1 및 MMP-9의 억제 효능을 측정하기 위해 유산균을 경구투여한 후 피부조직을 적출하여 Quantikine ELISA pro MMP-1 및 MMP-9 kit(R&D systems, Minneapolis, MN, USA)를 사용하여 MMP-1 및 MMP-9 단백질의 발현량을 측정하였다.MMP-1 and MMP-9 are proteins associated with skin aging and are known to be increased by oxidative stress such as ultraviolet rays, so in this experiment, we measured the inhibitory effect of MMP-1 and MMP-9 induced from oxidative damage. After oral administration of lactic acid bacteria, the skin tissue was extracted and the expression levels of MMP-1 and MMP-9 proteins were measured using Quantikine ELISA pro MMP-1 and MMP-9 kit (R&D systems, Minneapolis, MN, USA). .
그 결과, 도 1에 개시한 바와 같이 자외선을 조사하지 않은 정상군(Nor)에 비해, 자외선을 조사한 군(Veh)에서 MMP-1 및 MMP-9의 발현량이 현저하게 증가한 것을 확인하였고, 본 발명의 페디오코쿠스 이노피나투스 처리군에서 MMP-1 및 MMP-9의 발현량이 자외선 조사군에 비해 현저하게 감소하는 것을 확인하였다. As a result, it was confirmed that the expression levels of MMP-1 and MMP-9 were significantly increased in the ultraviolet-irradiated group (Veh) compared to the normal group (Nor) not irradiated with ultraviolet rays as disclosed in FIG. 1, and the present invention It was confirmed that the expression levels of MMP-1 and MMP-9 in the Pediococcus inofinatus treatment group were significantly reduced compared to the ultraviolet irradiation group.
실시예Example 3. 페디오코쿠스 이노피나투스 3. Pediococcus Inofinatus 의of 처리에 따른 피부조직의 변화 분석 Analysis of changes in skin tissue according to treatment
자외선으로 주름을 유도한 동물모델에서 무모생쥐군의 피부조직을 적출하여 H&E 염색 및 Masson's trichrome 염색을 하였다.Skin tissues of hairless mice were extracted from the animal model in which wrinkles were induced by UV rays, followed by H&E staining and Masson's trichrome staining.
도 2A에 개시한 H&E 염색 결과에서 보는 바와 같이, 자외선 조사군은 자외선을 처리하지 않은 대조군에 비해서, 각질층이 더 형성되었고, 표피의 두께도 증가된 것을 확인할 수 있었고, 페디오코쿠스 이노피나투스를 투여한 군에서는 각질층이 완화되었으며 표피두께가 감소한 것을 확인할 수 있었다. As can be seen from the H&E staining results disclosed in FIG. 2A, compared to the control group not treated with ultraviolet rays, the ultraviolet irradiation group had a more stratum corneum formed and the thickness of the epidermis was also increased, and Pediococcus inofinatus was In the administered group, it was confirmed that the stratum corneum was alleviated and the thickness of the epidermis was decreased.
도 2B에 개시한 Masson's trichome 염색결과에서도, 자외선을 조사한 군에서는 각질층이 굵게 형성되었으며, 콜라겐 섬유가 관찰되지 않았으나, 페디오코쿠스 이노피나투스를 투여한 군에서는 콜라겐 섬유가 자외선 조사군에 비해서 증가함을 확인하였다. In the result of Masson's trichome staining disclosed in FIG. 2B, the stratum corneum was formed thick in the group irradiated with ultraviolet rays, and collagen fibers were not observed, but in the group administered with Pediococcus inofinatus, the collagen fibers increased compared to the ultraviolet irradiated group. Was confirmed.
또한, H&E 염색을 실시한 조직의 케라틴층에서 표피 기저막까지의 두께를 현미경에 장착된 자를 이용하여 측정한 결과, 자외선 조사군에서 표피 두께가 증가되었고, 페디오코쿠스 이노피나투스를 투여한 군의 표피의 두께가 감소하였음을 확인하였다(도 2C). In addition, as a result of measuring the thickness from the keratin layer of the tissue subjected to H&E staining to the epidermal basement membrane using a ruler mounted on a microscope, the thickness of the epidermis was increased in the ultraviolet irradiation group, and the epidermis of the group administered Pediococcus inofinatus. It was confirmed that the thickness of the was decreased (FIG. 2C).
실시예Example 4. 4. 경피Transdermal 수분손실량( Moisture loss ( TEWLTEWL : : transepidermaltransepidermal water loss) 평가 및 분석 water loss) evaluation and analysis
경피수분 손실량은 항온 및 항습 조건(23℃, 상대습도 50%)에서 투와미터(Tewameter Courage & Khazaka, Germany) 장비를 이용하여 피부에서 증발하는 수분의 양을 면적과 시간에 따라 계산하여, 증발하는 수분의 양(g/m2/hr)을 보습 전자센서로 읽고, 이를 수치화하여 피부의 보습력을 측정하였다. 보습 효과를 확인하기 위하여 각 실험군의 무모 생쥐의 피부에서 경피 수분손실량 분석을 실시하였다.The amount of transdermal moisture loss is calculated by calculating the amount of moisture evaporated from the skin according to the area and time using the Tewameter Courage & Khazaka, Germany equipment under constant temperature and humidity conditions (23℃, 50% relative humidity), and evaporation. The moisture content (g/m 2 /hr) was read with a moisturizing electronic sensor, and it was quantified to measure the moisturizing power of the skin. In order to confirm the moisturizing effect, the percutaneous moisture loss was analyzed in the skin of hairless mice of each experimental group.
경피 수분손실량 수치는 피부로부터 발산되는 수분량으로, 수치가 높을수록 피부의 보습 기능이 떨어져 있음을 의미하며, 피부 고유의 장벽 기능이 손상되었음을 나타낸다.The transdermal moisture loss value is the amount of moisture emanated from the skin. The higher the value, the less moisturizing function of the skin is, indicating that the inherent barrier function of the skin is damaged.
그 결과, 도 3에 나타낸 바와 같이 자외선조사에 의해 피부 장벽이 손상되어 피부 보습 기능이 떨어진 것을 확인하였고(p<0.0001), 본 발명의 페디오코쿠스 이노피나투스를 처리하여 자외선에 의해 유발된 피부 수분 손실이 유의미하게 감소된 결과를 획득하였다(p<0.0001). As a result, as shown in FIG. 3, it was confirmed that the skin barrier was damaged by ultraviolet irradiation and the skin moisturizing function was degraded (p<0.0001), and the skin caused by ultraviolet rays was treated with Pediococcus Inofinatus of the present invention. The water loss was significantly reduced (p<0.0001).
실시예Example 5. 피부수분량(Capacitance) 측정 5. Measurement of skin moisture content (Capacitance)
피부수분량은 항온 및 항습 조건(23℃, 상대습도 50%)에서 코니오미터(Corneometer Courage & Khazaka, Germany) 장비를 이용하여 피부에 함유된 수분을 측정하였다. 피부의 표피 내에 존재하는 수분의 양은 센서를 이용하여 수분의 이온 정도를 측정하고, 이를 수치화하여 수분의 양을 계산함으로써 보습력을 측정하였다.The moisture content of the skin was measured using a Corneometer (Corneometer Courage & Khazaka, Germany) equipment under constant temperature and humidity conditions (23° C., 50% relative humidity). Moisturizing power was measured by measuring the amount of moisture in the skin's epidermis using a sensor, and calculating the amount of moisture by quantifying it.
도 4에 개시한 바와 같이, 본 발명의 페디오코쿠스 이노피나투스 투여군은 자외선 조사군에 비해서 수분함유량이 높게 나타났다. 이러한 결과를 토대로 본 발명의 페디오코쿠스 이노피나투스는 피부 보습효과가 있음을 확인할 수 있었다.As disclosed in FIG. 4, the group administered with Pediococcus inofinatus according to the present invention showed a higher water content than the ultraviolet irradiation group. Based on these results, it was confirmed that the Pediococcus inofinatus of the present invention has a skin moisturizing effect.
실시예Example 6. 히알루론산( 6. Hyaluronic acid ( HyaluronicHyaluronic Acid; HA)의 함량 변화 분석 Acid; HA) content change analysis
적출한 피부 조직을 PBS로 세척한 후, 100㎕의 PBS를 넣고 조직파쇄기(Precellys 24, Bertin technologies, Montigny-le-Bretonneux, France)로 갈아주었다. 이후, 100㎕의 lysis buffer (R&D systems, Minneapolis, MN, USA)를 첨가한 후 얼음에서 30분 동안 반응시켰다. 12000 rpm에서 10분 동안 원심분리하여 상층액을 -80℃에 보관하고 히알루론산 함량 분석에 사용하였다. 상층액에서 HA의 분석은 R&D System kit(R&D systems, Minneapolis, MN, USA)로 수행하였으며 마이크로플레이트 리더(Molecular Devices, LLC, Sunnyvale, CA, USA)로 540nm에서 흡광도를 측정하였다.After washing the extracted skin tissue with PBS, 100 µl of PBS was added and replaced with a tissue shredder (Precellys 24, Bertin technologies, Montigny-le-Bretonneux, France). Thereafter, 100 μl of lysis buffer (R&D systems, Minneapolis, MN, USA) was added and reacted on ice for 30 minutes. After centrifugation at 12000 rpm for 10 minutes, the supernatant was stored at -80°C and used for hyaluronic acid content analysis. Analysis of HA in the supernatant was performed with an R&D System kit (R&D systems, Minneapolis, MN, USA), and absorbance was measured at 540 nm with a microplate reader (Molecular Devices, LLC, Sunnyvale, CA, USA).
그 결과, 도 5에 개시한 바와 같이, 자외선 조사에 의해 히알루론산의 함량은 감소하였으나, 본 발명의 페디오코쿠스 이노피나투스를 처리한 실험군에서는 히알루론산의 함량이 증가한 것을 확인하였다.As a result, as disclosed in FIG. 5, the content of hyaluronic acid decreased by ultraviolet irradiation, but it was confirmed that the content of hyaluronic acid increased in the experimental group treated with Pediococcus inofinatus of the present invention.
[통계 처리][Statistics processing]
본 발명의 실시예에서 획득한 결과는 통계분석 프로그램(one-way ANOVA와 Student t-test)을 이용하여 대조군과 실시군 간의 유의성을 검정하였다. The results obtained in the examples of the present invention were tested for significance between the control group and the implementation group using a statistical analysis program (one-way ANOVA and Student t-test).
<110> Korea Institute of Oriental Medicine Korea Food Research Institute <120> Composition for improving skin wrinkle and skin moisturizing comprising Pediococcus inopinatus as effective component <130> PN19094 <160> 3 <170> KopatentIn 2.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> universal primer; 27F <400> 1 agagtttgat catggctcag 20 <210> 2 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> universal primer; 1492R <400> 2 ggataccttg ttacgactt 19 <210> 3 <211> 1359 <212> DNA <213> Pediococcus <400> 3 tggcgaacgg gtgagtaaca cgtgggtaac ctgcccagaa gtgggggata acacctggaa 60 acagatgcta ataccgcata ataaagtaaa ccgcatggtt ttcttttaaa agatggcttc 120 ggctatcact tctggatgga cccgcggcgt attagctagt tggtgagata aaggcccacc 180 aaggctgtga tacgtagccg acctgagagg gtaatcggcc acattgggac tgagacacgg 240 cccagactcc tacgggaggc agcagtaggg aatcttccac aatggacgaa agtctgatgg 300 agcaacgccg cgtgagtgat gaaggcttta gggtcgtaaa actctgttgt tggagaagaa 360 cgtgtgtgag agtaactgct catgcagtga cggtatccaa ccagaaagcc acggctaact 420 acgtgccagc agccgcggta atacgtaggt ggcaagcgtt atccggattt attgggcgta 480 aagcgagcgc aggcggtttc ttaagtctaa tgtgaaagcc ttcggcttaa ccgaagaagt 540 gcattggaaa ctgggaaact tgagtgcaga agaggacagt ggaactccat gtgtagcggt 600 gaaatgcgta gatatatgga agaacaccag tggcgaaggc ggctgtctgg tctgtaactg 660 acgctgaggc tcgaaagcat gggtagcgaa caggattaga taccctggta gtccatgccg 720 taaacgatga atgctaagtg ttggagggtt tccgcccttc agtgctgcag ctaacgcatt 780 aagcattccg cctggggagt acgaccgcaa ggttgaaact caaaagaatt gacgggggcc 840 cgcacaagcg gtggagcatg tggtttaatt cgaagctacg cgaagaacct taccaggtct 900 tgacatcttc tgctaaccta agagattagg cgttcccttc ggggacagaa tgacaggtgg 960 tgcatggttg tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgca acgagcgcaa 1020 cccctattat tagttgccag cattaagttg ggcactctag tgagactgcc ggtgataaac 1080 cggaggaagg tggggacgac gtcaaatcat catgcccctt atgacctggg ctacacacgt 1140 gctacaatgg acggtacaac gagttgcgag accgcgaggt taagctaatc tcttaaaacc 1200 gttctcagtt cggactgcag gctgcaactc gcctgcacga agttggaatc gctagtaatc 1260 gcggatcagc atgccgcggt gaatacgttc ccgggccttg tacacaccgc ccgtcacacc 1320 atgagagttt gtaacaccca aagccggtgg agtaacctt 1359 <110> Korea Institute of Oriental Medicine Korea Food Research Institute <120> Composition for improving skin wrinkle and skin moisturizing comprising Pediococcus inopinatus as effective component <130> PN19094 <160> 3 <170> KopatentIn 2.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> universal primer; 27F <400> 1 agagtttgat catggctcag 20 <210> 2 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> universal primer; 1492R <400> 2 ggataccttg ttacgactt 19 <210> 3 <211> 1359 <212> DNA <213> Pediococcus <400> 3 tggcgaacgg gtgagtaaca cgtgggtaac ctgcccagaa gtgggggata acacctggaa 60 acagatgcta ataccgcata ataaagtaaa ccgcatggtt ttcttttaaa agatggcttc 120 ggctatcact tctggatgga cccgcggcgt attagctagt tggtgagata aaggcccacc 180 aaggctgtga tacgtagccg acctgagagg gtaatcggcc acattgggac tgagacacgg 240 cccagactcc tacgggaggc agcagtaggg aatcttccac aatggacgaa agtctgatgg 300 agcaacgccg cgtgagtgat gaaggcttta gggtcgtaaa actctgttgt tggagaagaa 360 cgtgtgtgag agtaactgct catgcagtga cggtatccaa ccagaaagcc acggctaact 420 acgtgccagc agccgcggta atacgtaggt ggcaagcgtt atccggattt attgggcgta 480 aagcgagcgc aggcggtttc ttaagtctaa tgtgaaagcc ttcggcttaa ccgaagaagt 540 gcattggaaa ctgggaaact tgagtgcaga agaggacagt ggaactccat gtgtagcggt 600 gaaatgcgta gatatatgga agaacaccag tggcgaaggc ggctgtctgg tctgtaactg 660 acgctgaggc tcgaaagcat gggtagcgaa caggattaga taccctggta gtccatgccg 720 taaacgatga atgctaagtg ttggagggtt tccgcccttc agtgctgcag ctaacgcatt 780 aagcattccg cctggggagt acgaccgcaa ggttgaaact caaaagaatt gacgggggcc 840 cgcacaagcg gtggagcatg tggtttaatt cgaagctacg cgaagaacct taccaggtct 900 tgacatcttc tgctaaccta agagattagg cgttcccttc ggggacagaa tgacaggtgg 960 tgcatggttg tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgca acgagcgcaa 1020 cccctattat tagttgccag cattaagttg ggcactctag tgagactgcc ggtgataaac 1080 cggaggaagg tggggacgac gtcaaatcat catgcccctt atgacctggg ctacacacgt 1140 gctacaatgg acggtacaac gagttgcgag accgcgaggt taagctaatc tcttaaaacc 1200 gttctcagtt cggactgcag gctgcaactc gcctgcacga agttggaatc gctagtaatc 1260 gcggatcagc atgccgcggt gaatacgttc ccgggccttg tacacaccgc ccgtcacacc 1320 atgagagttt gtaacaccca aagccggtgg agtaacctt 1359
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