KR20180108209A - Composition for treatment of skin damage comprising tyndallized Lactobacillus rhamnosus JC1225 - Google Patents
Composition for treatment of skin damage comprising tyndallized Lactobacillus rhamnosus JC1225 Download PDFInfo
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- KR20180108209A KR20180108209A KR1020170037668A KR20170037668A KR20180108209A KR 20180108209 A KR20180108209 A KR 20180108209A KR 1020170037668 A KR1020170037668 A KR 1020170037668A KR 20170037668 A KR20170037668 A KR 20170037668A KR 20180108209 A KR20180108209 A KR 20180108209A
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Abstract
Description
락토바실러스 람노서스 JC1225 균주 또는 이의 사멸화된 균체를 유효성분으로 포함하는 피부 보습, 주름 개선, 또는 피부 미백 효과를 갖는 조성물에 관한 것이다.The present invention relates to a composition having skin moisturizing, wrinkle improving, or skin whitening effect, comprising Lactobacillus lambsosus JC1225 strain or its killed cell as an active ingredient.
락토바실러스 람노서스(Lactobacillusrhamnosus)는 유산균의 일종으로 정장 작용(intestinal regulation), 항균 작용, 아토피 예방, 및 체지방 저하 등 여러 효과를 가진 것으로 알려져 있다. 또한 근래에는 사멸화된 유산균에 대한 연구가 많이 진행되고 있는데, 사멸화된 유산균은 생균을 사멸화 시키는 과정에서 생겨난 세포벽 성분인 리포테이코산(Lipoteichoic acid) 등이 장벽에 부착하여 유해균 정착을 억제하고, 유산(lactic acid), 및 박테리오신(bacteriocin) 등의 발효물 내의 항균 성분이 직접 유해균에 작용하여 억제시킬 수 있는 등의 기능을 갖는 것으로 알려져 있다. 또한, 외부 세균등의 감염방어의 중요한 물질인 면역글로불린 A는 보통 유산균 생균에 의하기 보다는 사멸화된 유산균체들이 소장으로 흡수되어 대식세포를 자극하여 생산된다는 것과, 사멸화시킨 유산균은 생균의 먹이로 활용 되어 유산균의 정장 작용(intestinal regulation)에 사멸화 유산균이 우수한 프리바이오틱스(Prebiotics)로서 기능하는 것이 밝혀져 있다.Lactobacillus ramno suspension (Lactobacillusrhamnosus) is known to have several effects, such as suit acts as a kind of lactic acid bacteria (intestinal regulation), antibacterial action, atopic prevention, and decreased body fat. In recent years, many studies on killed lactic acid bacteria have been carried out. Lactic acid bacteria that have been killed have inhibited the fixation of harmful bacteria by attaching to the wall, such as Lipoteichoic acid, which is a cell wall component generated in the process of killing the live bacteria , Lactic acid, and bacteriocin can act directly on the harmful microorganisms and inhibit them. In addition, immunoglobulin A, which is an important substance of infection defense such as external bacteria, is usually produced by stimulating macrophages to be absorbed into the small intestine rather than by lactic acid bacteria, and the killed lactic acid bacteria are fed to live bacteria And it has been found that the killed lactic acid bacteria function as excellent prebiotics in the intestinal regulation of lactic acid bacteria.
사멸화 유산균의 면역 관련 기작에 관하여도 많은 연구가 진행되고 있는데, 사멸화 유산균의 면역 활성 메카니즘 논문 및 문헌으로는 Lactic Acid Bacteria - R & D for Food, Health and Livestock Purposes (2013) 등이 있고, 고분자 다당 바인더에 컨쥬게이트된 락토바실러스 람노서스 RHT-3201의 아토피 예방 및 치료에 대한 특허가 등록된 바 있다(대한민국 등록특허 제10-1640744호).There have been many studies on the immune-related mechanisms of the killed lactic acid bacteria. Lactic Acid Bacteria - R & D for Food, Health and Livestock Purposes (2013) A patent for the prevention and treatment of atopy of Lactobacillus lambosus RHT-3201 conjugated to a polymeric polysaccharide binder has been registered (Korean Patent No. 10-1640744).
이러한 배경하에서, 보다 유용한 사멸화 유산균 균주 개발과 더불어 기존에 알려지지 않은 피부 보습, 주름 개선, 미백 기능성 등으로서 이용하기 위한 사멸화 유산균의 개발이 요구되고 있다.Under these circumstances, there is a demand for the development of more suicide lactobacillus strains that are more useful, as well as the development of a suicide lactic acid bacterium for use as skin moisturizing, wrinkle improvement, whitening function, and the like.
일 양상은 피부 손상 개선 효과를 가지는 락토바실러스 람노서스 JC1225 균주를 제공한다.One aspect provides Lactobacillus laminosus JC1225 strain having skin damaging improving effect.
다른 양상은 락토바실러스 람노서스 JC1225 균주를 포함하는 배양물을 제공한다.Another aspect provides a culture comprising Lactobacillus laminocus JC1225 strain.
다른 양상은 락토바실러스 람노서스 JC1225 균주를 유효성분으로 포함하는 피부 손상을 치료하기 위한 약학적 조성물을 제공한다.Another aspect provides a pharmaceutical composition for treating skin damage comprising Lactobacillus rhamnosus JC1225 strain as an active ingredient.
다른 양상은 락토바실러스 람노서스 JC1225 균주를 유효성분으로 포함하는 피부 손상을 개선 또는 예방하기 위한 화장료 조성물을 제공한다.Another aspect provides a cosmetic composition for improving or preventing skin damage comprising Lactobacillus lambosus JC1225 strain as an active ingredient.
다른 양상은 락토바실러스 람노서스 JC1225 균주를 유효성분으로 포함하는 피부 손상을 개선 또는 예방하기 위한 건강기능식품용 조성물을 제공한다.Another aspect provides a composition for a health functional food for improving or preventing skin damage comprising Lactobacillus lambosus JC1225 strain as an active ingredient.
다른 양상은 락토바실러스 람노서스 JC1225 균주의 사멸화 균체 및 이를 제조하는 방법을 제공한다.Another aspect provides a killed cell of a Lactobacillus laminocus JC1225 strain and a method of producing the same.
일 양상은 피부 손상 개선 효과를 가지는 락토바실러스 람노서스 JC1225 균주(Lactobacillus rhamnosus JC1225 strain) 및 이를 포함하는 배양물를 제공한다.One aspect provides a Lactobacillus rhamnosus JC1225 strain and a culture comprising the Lactobacillus rhamnosus JC1225 strain having skin damaging improving effect.
상기 락토바실러스 람노서스 JC1225 균주는, 수탁번호 KCTC 13184BP의 미생물 및 이와 동일한 유전자를 갖거나, 이로부터 유래한 균주로 분리되는 미생물을 포함한다. 여기서 "유래한 균주"는 특정 균주가 JC1225 균주의 생식으로 발생하였거나, 또는 JC1225 균주의 유전자 정보를 이용하여 당업계에서 통상적인 기술을 사용하여 유전자 조작으로 발생시킨 균주로서, JC1225와 동일 또는 유사한 미생물학적 특성을 갖는 균주를 포함하는 의미로 사용된다. 상기 JC1225 균주는 서열번호 1의 16S rDNA 서열을 갖거나, 혹은 이와 97% 이상의 유사성(similarity)을 갖는 16S rDNA 서열을 갖는 것일 수 있다. The Lactobacillus lambusus JC1225 strain includes a microorganism having the accession number KCTC 13184BP and a microorganism having the same gene or separated into a strain derived therefrom. Herein, the term "derived strain" refers to a strain generated by genetically engineering a strain of JC1225, or using the gene information of JC1225 strain, by genetic engineering using a technique common in the art, and a microorganism identical or similar to JC1225 Is used in the sense of including a strain having a characteristic of a microorganism. The JC1225 strain may have a 16S rDNA sequence of SEQ ID NO: 1, or a 16S rDNA sequence having 97% or more similarity thereto.
본 명세서에서 용어 "유전자 조작 (genetic engineering)" 또는 "유전자 재조합 (genetic recombination)"은 세포에 대하여 하나 이상의 유전적 변형 (genetic modification)을 도입하는 행위 또는 그에 의하여 만들어진 세포를 나타낸다. 예를 들면, 유전자 조작으로 발생된 균주는 락토바실러스 람노서스 JC1225 균주의 피부 손상 개선 효과를 갖는 유전자의 발현 또는 활성이 증가되도록 유전적으로 조작된 것, 예를 들면 외인성 락토바실러스 람노서스 JC1225 균주 유전자를 포함하는 것일 수 있다. 상기 활성 증가는 주어진 유전적으로 조작되지 않은 모세포(예, 야생형)가 갖지 않는 또는 갖는 내재적 단백질 또는 효소의 활성에 비해, 동일한 타입의 단백질 또는 효소의 활성이 보다 더 높은 활성을 갖는 것을 의미할 수 있다. 상기 외인성 유전자는, 상기 균주에서 그 모세포에 비하여 언급된 단백질의 활성이 증가되기에 충분한 양으로 발현된 것일 수 있다. 상기 외인성 유전자는 발현 벡터를 통하여 세포 내로 도입된 것일 수 있다. 또한, 상기 외인성 유전자는 선형 폴리뉴클레오티드 형태로 세포 내로 도입된 것일 수 있다. 또한, 상기 외인성 유전자는 세포 내에서 발현 벡터 (예, 플라스미드)로부터 발현되는 것일 수 있다. 또한, 상기 외인성 유전자는 안정적인 발현을 위하여 세포 내의 유전물질 (예, 염색체)에 삽입되어 발현되는 것일 수 있다. 상기 외인성 유전자가 도입되는 세포는 대장균, 효모, 또는 동물 세포 등의 이종 세포를 포함하며, 상업적으로 구매 가능한 것일 수 있다.The term "genetic engineering" or "genetic recombination" as used herein refers to the act of introducing one or more genetic modifications to a cell or cells made thereby. For example, a strain generated by genetic engineering is genetically engineered to increase the expression or activity of a gene having an effect of improving skin damage of Lactobacillus lambusus JC1225 strain, for example, an exogenous Lactobacillus lambosus JC1225 strain gene May include. This increase in activity may mean that the activity of the same type of protein or enzyme has a higher activity than the activity of an endogenous protein or enzyme that does not or does not have a given genetically engineered parental cell (e. G., Wild type) . The exogenous gene may be expressed in an amount sufficient to increase the activity of the protein mentioned in the strain relative to the parental cell. The exogenous gene may be introduced into a cell through an expression vector. In addition, the exogenous gene may be introduced into cells as a linear polynucleotide form. In addition, the exogenous gene may be expressed from an expression vector (e.g., a plasmid) in a cell. In addition, the exogenous gene may be inserted into a genetic material (e.g., a chromosome) in a cell for stable expression. The cell to which the exogenous gene is introduced includes heterologous cells such as Escherichia coli, yeast, or animal cells, and may be commercially available.
용어 "벡터(vector)"는 숙주 세포에서 목적 유전자를 발현시키기 위한 수단을 의미하며 유전자 재조합을 통해 원하는 목적 유전자를 포함한다. 예를 들어, 플라스미드 벡터, 코즈미드 벡터 및 박테리오파아지 벡터, 아데노바이러스 벡터, 레트로바이러스 벡터 및 아데노-연관 바이러스 벡터와 같은 바이러스 벡터를 포함한다. 재조합 벡터로 사용될 수 있는 벡터는 당업계에서 종종 사용되는 플라스미드 (예를 들면, pSC101, pGV1106, pACYC177, ColE1, pKT230, pME290, pBR322, pUC8/9, pUC6, pBD9, pHC79, pIJ61, pLAFR1, pHV14, pGEX 시리즈, pET 시리즈 및 pUC19 등), 파지 (예를 들면, λgt4λB, λ-Charon, λΔz1 및 M13 등) 또는 바이러스 (예를 들명, SV40 등)를 조작하여 제작될 수 있다.The term "vector" means means for expressing a gene of interest in a host cell and includes the desired gene of interest through gene recombination. For example, viral vectors such as plasmid vectors, cosmid vectors and bacteriophage vectors, adenovirus vectors, retroviral vectors, and adeno-associated viral vectors. The vector which can be used as a recombinant vector may be a plasmid (for example, pSC101, pGV1106, pACYC177, ColE1, pKT230, pME290, pBR322, pUC8 / 9, pUC6, pBD9, pHC79, pIJ61, pLAFR1, pHV14, pGEX series, pET series, and pUC19), phage (e.g.,? gt4? B,? -charon,?? z1 and M13) or viruses (e.g., SV40 and the like).
상기 재조합 벡터에서 상기 단백질 복합체를 코딩하는 폴리뉴클레오타이드는 프로모터에 작동적으로 연결될 수 있다. 용어 "작동 가능하게 연결된(operatively linked)"은 뉴클레오타이드 발현 조절 서열(예를 들어, 프로모터 서열)과 다른 뉴클레오타이드 서열 사이의 기능적인 결합을 의미한다. 상기 조절 서열은 "작동 가능하게 연결(operatively linked)"됨으로써 다른 뉴클레오타이드 서열의 전사 및/또는 해독을 조절할 수 있다.The polynucleotide encoding the protein complex in the recombinant vector may be operatively linked to a promoter. The term "operatively linked" refers to the functional linkage between a nucleotide expression control regulatory sequence (e.g., a promoter sequence) and another nucleotide sequence. The regulatory sequence may be "operatively linked" to regulate transcription and / or translation of other nucleotide sequences.
상기 재조합 벡터는, 전형적으로 클로닝을 위한 벡터 또는 발현을 위한 벡터로서 구축될 수 있다. 상기 발현용 벡터는 당업계에서 식물, 동물 또는 미생물에서 외래의 단백질을 발현하는 데 사용되는 통상의 것을 사용할 수 있다. 상기 재조합 벡터는 당업계에 공지된 다양한 방법을 통해 구축될 수 있다.The recombinant vector can typically be constructed as a vector for cloning or as a vector for expression. The expression vector may be any conventional vector used in the art to express an exogenous protein in plants, animals or microorganisms. The recombinant vector may be constructed by a variety of methods known in the art.
상기 재조합 벡터는 원핵 세포 또는 진핵 세포를 숙주로 하여 구축될 수 있다. 예를 들어, 사용되는 벡터가 발현 벡터이고, 원핵 세포를 숙주로 하는 경우에는, 전사를 진행시킬 수 있는 강력한 프로모터 (예를 들어, pLλ 프로모터, CMV 프로모터, trp 프로모터, lac 프로모터, tac 프로모터, T7 프로모터 등), 해독의 개시를 위한 라이보좀 결합 자리 및 전사/해독 종결 서열을 포함하는 것이 일반적이다. 진핵 세포를 숙주로 하는 경우에는, 벡터에 포함되는 진핵 세포에서 작동하는 복제원점은 f1 복제원점, SV40 복제원점, pMB1 복제원점, 아데노 복제원점, AAV 복제원점 및 BBV 복제원점 등을 포함하나, 이에 한정되는 것은 아니다. 또한, 포유동물 세포의 게놈으로부터 유래된 프로모터 (예를 들어, 메탈로티오닌 프로모터) 또는 포유동물 바이러스로부터 유래된 프로모터 (예를 들어, 아데노바이러스 후기 프로모터, 백시니아 바이러스 7.5K 프로모터, SV40 프로모터, 사이토메갈로바이러스 프로모터 및 HSV의 tk프로모터)가 이용될 수 있으며, 전사 종결 서열로서 폴리아데닐화 서열을 일반적으로 갖는다.The recombinant vector may be constructed with prokaryotic or eukaryotic cells as hosts. For example, when the vector used is an expression vector and the prokaryotic cell is used as a host, a strong promoter capable of promoting transcription (for example, pL? Promoter, CMV promoter, trp promoter, lac promoter, tac promoter, T7 Promoter, etc.), a ribosome binding site for initiation of translation, and a transcription / translation termination sequence. When a eukaryotic cell is used as a host, the origin of replication that functions in eukaryotic cells contained in the vector includes f1 replication origin, SV40 replication origin, pMB1 replication origin, adeno replication origin, AAV replication origin, and BBV replication origin But is not limited thereto. Also, promoters derived from the genome of mammalian cells (e.g., metallothionein promoter) or mammalian viruses (e.g., adenovirus late promoter, vaccinia virus 7.5K promoter, SV40 promoter, The cytomegalovirus promoter and the tk promoter of HSV) can be used and generally have a polyadenylation sequence as a transcription termination sequence.
상기 재조합 세포 또는 재조합 균주는 상기 재조합 벡터를 적절한 숙주 세포에 도입시킴으로써 얻어진 것일 수 있다. 상기 숙주 세포는 상기 재조합 벡터를 안정되면서 연속적으로 클로닝 또는 발현시킬 수 있는 세포로서 당업계에 공지된 어떠한 숙주 세포도 이용할 수 있으며, 원핵 세포로는, 예를 들어, E. coliJM109, E. coliBL21, E. coli RR1, E. coli LE392, E. coli B, E. coli X 1776, E. coli W3110, 바실러스 서브틸리스, 바실러스 츄린겐시스와 같은 바실러스 속 균주, 그리고 살모넬라 티피무리움, 세라티아 마르세슨스 및 다양한 슈도모나스 종과 같은 장내균과 균주 등이 있으며, 진핵 세포에 형질 전환시키는 경우에는 숙주 세포로서, 효모(Saccharomyce cerevisiae), 곤충 세포, 식물 세포 및 동물 세포, 예를 들어, Sp2/0, CHO(Chinese hamster ovary) K1, CHO DG44, PER.C6, W138, BHK, COS-7, 293, HepG2, Huh7, 3T3, RIN, MDCK 세포주 등이 이용될 수 있다. The recombinant cell or the recombinant strain may be obtained by introducing the recombinant vector into an appropriate host cell. The host cell may be any host cell known in the art as a cell capable of continuously cloning or expressing the recombinant vector stably. Examples of the prokaryotic cell include E. coli JM109, E. coli BL21, Bacillus strains such as E. coli RR1, E. coli LE392, E. coli B, E. coli X1776, E. coli W3110, Bacillus subtilis, and Bacillus turginensis, and Salmonella typhimurium and Serratia marsea Such as yeast (Saccharomyce cerevisiae), insect cells, plant cells and animal cells, for example, Sp2 / 0, CHO (Chinese hamster ovary) K1, CHO DG44, PER.C6, W138, BHK, COS-7, 293, HepG2, Huh7, 3T3, RIN and MDCK cell lines.
상기 락토바실러스 람노서스 JC1225 균주는 피부 손상 개선 또는 치료 효과를 갖는다. 용어 "피부 손상"은 외부의 물리적 손상, 화학 물질, 세균, 곰팡이, 바이러스 등의 침투, 자외선에 대한 노출, 피부의 수분 손실, 노화 등에 의한 주름, 및 피부 색소 침착, 피부 염증, 지루성 피부염, 붉어짐(발적), 태선화, 습진, 소양감, 및 아토피 피부염 등을 포함하는 피부 조직 또는 세포의 임상적 및 미용적 관점에서의 손상을 포함한다. 상기 피부는 얼굴, 손, 팔, 다리, 발, 가슴, 배, 등, 엉덩이, 및 두피를 포함하는 신체의 모든 피부 부위를 포함한다. 본 발명자는 락토바실러스 람노서스 JC1225 균주를 이용한 실험에서 상기 균주의 피부 보습, 주름 개선 및 피부 미백 효과를 확인하였으므로, 락토바실러스 람노서스 JC1225 균주는 피부 손상을 개선 또는 치료하는데 이용할 수 있다. 또한, 본 발명자는 락토바실러스 람노서스 JC1225 균주를 환자에게 투여하는 임상 시험에서 두피, 얼굴 및 등 피부 등의 피부 염증을 현저히 완화 내지 치료하는 효과를 확인하였으므로, 두피를 포함하는 피부의 피부 염증을 개선 또는 치료하는데 이용할 수 있다.The Lactobacillus laminosis JC1225 strain has an effect of improving or treating skin damage. The term "skin damage" refers to any type of skin damage, including, but not limited to, external physical damage, penetration of chemicals, bacteria, fungi, viruses, exposure to ultraviolet light, skin moisture loss, wrinkles due to aging and skin pigmentation, Including, but not limited to, skin or tissue damage including, but not limited to, skin irritation (redness), firming, eczema, pruritus, and atopic dermatitis. The skin includes all parts of the body including the face, hands, arms, legs, feet, chest, belly, back, buttocks, and scalp. The present inventor has confirmed the skin moisturizing, wrinkle improvement and skin whitening effect of the above strain in an experiment using Lactobacillus lambosus JC1225 strain, so that Lactobacillus lambosus JC1225 strain can be used for improving or treating skin damage. In addition, the inventor of the present invention confirmed that the clinical test of administering the Lactobacillus lambosus JC1225 strain to the patient significantly alleviated or treated the skin inflammation of the scalp, face, and back skin, thereby improving the skin inflammation of the skin including the scalp Or treatment.
상기 락토바실러스 람노서스 JC1225 균주는 사멸화된 것일 수 있다. 상기 사멸화는 당업계에서 알려진 틴달화(Tyndallized) 방법을 이용하여 수행할 수 있다. 일반적인 틴달화 방법은 균체가 내포하고 있는 기능성 성분이 변질되지 않는 50℃ 내지 180℃의 온도에서 15분 내지 120분간 미생물 배양체를 가열하는 단계, 및 상기 가열된 미생물 배양체를 10℃ 내지 40℃의 온도에서 냉각시키는 단계를 포함한다. 이후 수득한 미생물 배양체(사균체)는 동결 건조될 수 있다. 상기 미생물 배양체는 미생물 발효물일 수 있다. 상기 틴달화는 상기 미생물 발효물을 균체와 발효여액으로 분리한 후 각각 틴달화를 수행하는 것일 수 있다. 통상적으로는, 사멸화(틴딜화)할 때 배양액을 원심분리를 통하여 발효여액과 균체로 분리하여 각각 따로따로 사멸화(틴딜화)하여 건조하는 방법이 이용된다. 이후 상기 건조된 각각의 발효여액 및 균체를 혼합하는 것이 일반적이다. 그러나 이러한 방법은 배양액을 균체와 배양여액으로 원심분리하는 과정에서 배양액이 함유하고 있는 성분 중 인체에 유익한 박테리오신, 유기산, 아미노산, 인산염, 젖당, 및 펩타이드 등이 유실되는 단점을 가지고 있다. 따라서, 상기 틴달화는 균체 및 배양액(발효액)을 분리하지 않고 배지에서 배양한 배양물을 그대로 이용하여 수행하는 것일 수 있다. 배양물을 그대로 이용하여 틴달화를 수행한 경우, 이후 감압 농축 및 유동층 건조방법 등 당업계에서 통상적으로 이용되는 방식으로 건조하는 단계를 추가로 포함할 수 있다. 상기 배양물은 유산균을 배양하는데 통상적으로 이용되는 성분을 포함하고 있을 수 있으며, 상업적으로 구매가능한 것일 수 있다. 예를 들면, 일반적으로 사용하는 기본 배지(MRS배지)에 계면활성제, 단백질, 전분, 과당, 및 무기물 등을 첨가한 배양 배지를 사용하여 pH를 6.0~7.5으로 유지하면서 유산균을 배양할 수 있다.The Lactobacillus laminosus JC1225 strain may be killed. The killing may be performed using a Tyndallized method known in the art. The general method for culturing a microorganism is to heat the microorganism culture for 15 to 120 minutes at a temperature of 50 to 180 DEG C at which the functional ingredient contained in the microorganism is not altered, and heating the microorganism culture to a temperature of 10 to 40 DEG C . The resulting microorganism cultures (dead cells) can be lyophilized. The microorganism culture may be a microorganism fermentation product. The thymarization may be performed by separating the microbial fermentation product with the microbial cells and the fermentation filtrate, respectively, and then performing the tincturation. Normally, when the fermentation broth is killed (tin-dilution), the culture broth is separated into fermentation filtrate and microbial cells through centrifugation, and then each of them is separately killed (tin dilution) and dried. Thereafter, the dried fermentation filtrate and the cells are generally mixed. However, this method has disadvantages that bacteriocins, organic acids, amino acids, phosphates, lactose, and peptides, which are beneficial to the human body, are lost during the centrifugation of the culture solution with the culture solution. Therefore, the cultivation may be carried out using the culture obtained by culturing in the medium without separating the cells and the culture medium (fermentation broth). In the case where the cultivation is carried out using the culture as it is, it may further include a step of drying in a manner commonly used in the art such as a method of concentration under reduced pressure and a fluidized bed drying method. The culture may contain components conventionally used for culturing the lactic acid bacteria, and may be commercially available. For example, a lactic acid bacterium can be cultured while maintaining the pH at 6.0 to 7.5 using a culture medium supplemented with a surfactant, protein, starch, fructose, and an inorganic substance, etc. in a commonly used basic medium (MRS medium).
상기 락토바실러스 람노서스 JC1225 균주를 포함하는 배양물은 생균 또는 사멸화 락토바실러스 람노서스 JC1225 균체를 108 CFU/g 내지 1010 CFU/g 농도로 포함하는 것일 수 있다.The culture containing the Lactobacillus lambosus JC1225 strain may contain live cells or killed Lactobacillus lanxosus JC1225 cells at a concentration of 10 8 CFU / g to 10 10 CFU / g.
상기한 바와 같이 본 발명은 또한 락토바실러스 람노서스 JC1225 균주의 사멸화 균체를 제조하는 방법을 제공한다.As described above, the present invention also provides a method for producing the killed bacteria of Lactobacillus lambusus JC1225 strain.
다른 양상은 락토바실러스 람노서스 JC1225 균주를 유효성분으로 포함하는 피부 손상을 치료하기 위한 약학적 조성물을 제공한다.Another aspect provides a pharmaceutical composition for treating skin damage comprising Lactobacillus rhamnosus JC1225 strain as an active ingredient.
용어 "치료"는 질환, 장애 또는 병태, 또는 그의 하나 이상의 증상의 경감, 진행 억제 또는 예방을 지칭하거나, 그를 포함하며, "유효성분" 또는 "약제학적 유효량"은 질환, 장애 또는 병태, 또는 그의 하나 이상의 증상의 경감, 진행 억제 또는 예방에 충분한 본원에서 제공되는 발명을 실시하는 과정에서 이용되는 조성물의 임의의 양을 의미할 수 있다.Refers to or includes the reduction, progression, or prevention of a disease, disorder or condition, or one or more symptoms thereof, wherein the "active ingredient" or "pharmaceutically effective amount" refers to a disease, disorder or condition, May refer to any amount of composition used in the practice of the invention provided herein to ameliorate, inhibit or prevent progression of one or more symptoms.
일 구체예에 따른 상기 조성물은 조성물 총 중량에 대하여 0.001 중량% 내지 80 중량%의 락토바실러스 람노서스 JC1225 균주를 포함할 수 있다. 또한, 락토바실러스 람노서스 JC1225 균주의 투여 용량은 0.01mg 내지 10,000mg, 0.1mg 내지 1000mg, 1mg 내지 100mg, 0.01mg 내지 1000mg, 0.01mg 내지 100mg, 0.01mg 내지 10mg, 또는 0.01mg 내지 1mg일 수 있다. 또한, 락토바실러스 람노서스 JC1225 균주의 투여 용량은 1.0× 105 내지 1.0× 108 세포/kg(체중)일 수 있다. 다만, 투여량은 제제화 방법, 투여 방식, 환자의 연령, 체중, 성별, 병적 상태, 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하게 처방될 수 있고, 당업자라면 이러한 요인들을 고려하여 투여량을 적절히 조절할 수 있다. 투여 횟수는 1회 또는 임상적으로 용인 가능한 부작용의 범위 내에서 2회 이상이 가능하고, 투여 부위에 대해서도 1개소 또는 2개소 이상에 투여할 수 있다. 인간 이외의 동물에 대해서도, kg당 인간과 동일한 투여량으로 하거나, 또는 예를 들면 목적의 동물과 인간과의 기관(심장 등)의 용적비(예를 들면, 평균값) 등으로 상기의 투여량을 환산한 양을 투여할 수 있다. 가능한 투여 경로에는 경구, 설하, 비경구 (예를 들어, 피하, 근육내, 동맥내, 복강내, 경막내, 또는 정맥내), 직장, 국소 (경피 포함), 흡입, 및 주사, 또는 이식성 장치 또는 물질의 삽입을 포함할 수 있다. 일 구체예에 따른 치료의 대상동물로서는, 인간 및 그 밖의 목적으로 하는 포유동물을 예로 들 수 있고, 구체적으로는 인간, 원숭이, 마우스, 래트, 토끼, 양, 소, 개, 말, 돼지 등이 포함된다. 일 실시예에 따르면 상기 조성물은 사멸화 건조 균주를 포함하며, 1회 1g 내지 10g, 0.5g 내지 1.5g, 2.5g 내지 3.5g, 또는 4.5g 내지 5.5g 투여 가능하고, 1일 1회 내지 3회 투여될 수 있다.The composition according to one embodiment may comprise from 0.001% to 80% by weight of Lactobacillus laminosus JC1225 strain relative to the total weight of the composition. The dose of the Lactobacillus laminocus JC1225 strain may be 0.01 mg to 10,000 mg, 0.1 mg to 1000 mg, 1 mg to 100 mg, 0.01 mg to 1000 mg, 0.01 mg to 100 mg, 0.01 mg to 10 mg, or 0.01 mg to 1 mg . In addition, the dose of Lactobacillus laminosis JC1225 strain may be 1.0 x 10 5 to 1.0 x 10 8 cells / kg (body weight). However, the dose may be variously prescribed depending on factors such as the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, route of administration, excretion rate and responsiveness of the patient, These factors can be taken into account to appropriately adjust the dosage. The number of administrations may be one or two or more times within the range of clinically acceptable side effects, and the administration site may be administered at one site or two or more sites. For an animal other than a human, the dosage may be the same as that of a human per kg, or the dosage may be converted into a dose in terms of a volume ratio (for example, an average value) One dose may be administered. Possible routes of administration include oral, sublingual, parenteral (e.g. subcutaneous, intramuscular, intraarterial, intraperitoneal, intrathecal, or intravenous), rectal, topical (including transdermal), inhalation, Or insertion of a substance. Examples of animals to be treated in accordance with one embodiment include humans and other mammals for the purpose of administration. Specifically, humans, monkeys, mice, rats, rabbits, sheep, cows, dogs, horses, . According to one embodiment, the composition comprises an autotrophic dry strain and may be administered in an amount of 1 g to 10 g, 0.5 g to 1.5 g, 2.5 g to 3.5 g, or 4.5 g to 5.5 g, ≪ / RTI >
일 구체예에 따른 약학적 조성물은 약학적으로 허용가능한 담체 및/또는 첨가물을 포함할 수 있다. 예를 들어, 멸균수, 생리식염수, 관용의 완충제(인산, 구연산, 그 밖의 유기산 등), 안정제, 염, 산화방지제(아스코르브산 등), 계면활성제, 현탁제, 등장화제, 또는 보존제 등을 포함할 수 있다. 국소 투여를 위해, 생체고분자(biopolymer) 등의 유기물, 하이드록시아파타이트 등의 무기물, 구체적으로는 콜라겐 매트릭스, 폴리락트산 중합체 또는 공중합체, 폴리에틸렌글리콜 중합체 또는 공중합체 및 그의 화학적 유도체 등과 조합시키는 것도 포함할 수 있다. 일 구체예에 따른 약학적 조성물이 주사에 적당한 제형으로 조제되는 경우에는, 락토바실러스 람노서스 JC1225 균체가 약학적으로 허용가능한 담체 중에 용해 또는 분산되어 있거나 또는 용해 또는 분산되어 있는 용액 상태로 동결된 것일 수 있다. The pharmaceutical compositions according to one embodiment may comprise pharmaceutically acceptable carriers and / or additives. Examples thereof include sterilized water, physiological saline, buffering agents (phosphoric acid, citric acid, other organic acids, etc.), stabilizers, salts, antioxidants (such as ascorbic acid), surfactants, suspending agents, isotonic agents, can do. For the topical administration, organic substances such as biopolymers, and inorganic substances such as hydroxyapatite, specifically, collagen matrix, polylactic acid polymer or copolymer, polyethylene glycol polymer or copolymer, and chemical derivatives thereof . When a pharmaceutical composition according to one embodiment is formulated into a formulation suitable for injection, the Lactobacillus lanxosus JC1225 cells are dissolved or dispersed in a pharmaceutically acceptable carrier or frozen in a solution state in which they are dissolved or dispersed .
일 구체예에 따른 약학적 조성물은 그 투여방법이나 제형에 따라 필요한 경우, 현탁제, 용해보조제, 안정화제, 등장화제, 보존제, 흡착방지제, 계면활성화제, 희석제, 부형제, pH 조정제, 무통화제, 완충제, 환원제, 산화방지제 등을 적절히 포함할 수 있다. 상기에 예시된 것들을 비롯하여 본 발명에 적합한 약학적으로 허용되는 담체 및 제제는 문헌[Remington's Pharmaceutical Sciences, 19th ed., 1995]에 상세히 기재되어 있다. 일 구체예에 따른 약학적 조성물은 당해 발명이 속하는 기술 분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있는 방법에 따라, 약학적으로 허용되는 담체 및/또는 부형제를 이용하여 제제화함으로써 단위 용량 형태로 제조되거나 또는 다용량 용기 내에 내입시켜 제조될 수 있다. 이때 제형은 오일 또는 수성 매질 중의 용액, 현탁액 또는 유화액 형태이거나 분말, 과립, 정제 또는 캡슐 형태일 수 있다. The pharmaceutical composition according to one embodiment of the present invention may be formulated into various forms such as suspensions, solubilizers, stabilizers, isotonizing agents, preservatives, adsorption inhibitors, surfactants, diluents, excipients, pH adjusters, Buffers, reducing agents, antioxidants, and the like. Pharmaceutically acceptable carriers and formulations suitable for the present invention, including those exemplified above, are described in detail in Remington's Pharmaceutical Sciences, 19th ed., 1995. The pharmaceutical composition according to one embodiment of the present invention may be formulated into pharmaceutically acceptable carriers and / or excipients in accordance with a method which can be easily carried out by those skilled in the art, Or may be manufactured by penetration into a multi-dose container. The formulations may be in the form of solutions, suspensions or emulsions in oil or aqueous media, or in the form of powders, granules, tablets or capsules.
또한 상기 약학적 조성물은 피부에 도포하기 위한 피부외용제일 수 있다. 상기 피부외용제는 크림, 겔, 연고, 피부 유화제, 피부 현탁액, 경피전달성 패치, 약물 함유 붕대, 로션, 또는 그 조합일 수 있다. 상기 피부외용제는 통상 화장품이나 의약품 등의 피부외용제에 사용되는 성분, 예를 들면 수성성분, 유성성분, 분말성분, 알코올류, 보습제, 증점제, 자외선흡수제, 미백제, 방부제, 산화방지제, 계면활성제, 향료, 색제, 각종 피부 영양제, 또는 이들의 조합과 필요에 따라서 적절하게 배합될 수 있다. 상기 피부외용제는, 에데트산이나트륨, 에데트산삼나트륨, 시트르산나트륨, 폴리인산나트륨, 메타인산나트륨, 글루콘산 등의 금속봉쇄제, 카페인, 탄닌, 벨라파밀, 감초추출물, 글라블리딘, 칼린의 과실의 열수추출물, 각종 생약, 아세트산토코페롤, 글리틸리틴산, 트라넥삼산 및 그 유도체 또는 그 염등의 약제, 비타민 C, 아스코르브산인산마그네슘, 아스코르브산글루코시드, 알부틴, 코지산, 글루코스, 프룩토스, 트레할로스 등의 당류도 적절하게 배합할 수 있다.The pharmaceutical composition may also be a skin external preparation for application to the skin. The external preparation for skin may be a cream, a gel, an ointment, a skin emulsifier, a skin suspension, a transdermal patch, a drug-containing bandage, a lotion, or a combination thereof. The external preparation for skin is usually used as a component used in external skin preparations such as cosmetics or medicines such as an aqueous component, an oily component, a powder component, an alcohol, a moisturizer, a thickener, an ultraviolet absorber, a whitening agent, an antiseptic, , Coloring agents, various skin nutrients, or a combination thereof, and may be suitably blended as necessary. The external preparation for skin may be a metal blocker such as sodium edetate, sodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate or gluconic acid, caffeine, tannin, bellapamil, licorice extract, glabridine, Vitamin C, ascorbic acid magnesium phosphate, ascorbic acid glucoside, arbutin, kojic acid, glucose, fructose, fructose, fructose and other herbal medicines, various herbal medicines, tocopherol acetate, glycyrrhizic acid, Saccharides such as trehalose can also be appropriately compounded.
다른 양상은 락토바실러스 람노서스 JC1225 균주를 유효성분으로 포함하는 피부 손상을 개선 또는 예방하기 위한 화장료 조성물을 제공한다. Another aspect provides a cosmetic composition for improving or preventing skin damage comprising Lactobacillus lambosus JC1225 strain as an active ingredient.
상기 화장료 조성물은 국소 적용에 적합한 모든 제형으로 제공될 수 있다. 예를 들면, 용액, 수상에 유상을 분산시켜 얻은 에멀젼, 유상에 수상을 분산시켜 얻은 에멀젼, 현탁액, 고체, 겔, 분말, 페이스트, 마스크 팩 또는 시트, 포말(foam) 또는 에어로졸 조성물의 제형으로 제공될 수 있다. 이러한 제형의 조성물은 당해 분야의 통상적인 방법에 따라 제조될 수 있다.The cosmetic composition may be provided in all formulations suitable for topical application. For example, as a solution, an emulsion obtained by dispersing an oil phase in an aqueous phase, an emulsion obtained by dispersing an oil phase in water, a suspension, a solid, a gel, a powder, a paste, a mask pack or a sheet, a foam or an aerosol composition . Compositions of such formulations may be prepared according to conventional methods in the art.
상기 화장료 조성물은 보습제, 에몰리언트제, 자외선 흡수제, 방부제, 살균제, 산화 방지제, pH 조정제, 유기 및 무기 안료, 향료, 냉감제 또는 제한제를 더 포함할 수 있다. 상기 보습제 등의 추가 성분의 배합량은 본 발명의 목적 및 효과를 손상시키지 않는 범위 내에서 당업자가 용이하게 선정 가능하며, 그 배합량은 조성물 전체 중량을 기준으로 0.01 내지 5 중량%, 구체적으로 0.01 내지 3 중량%일 수 있다.The cosmetic composition may further comprise a moisturizing agent, an emollient agent, an ultraviolet absorber, an antiseptic, a bactericide, an antioxidant, a pH adjuster, an organic and inorganic pigment, a perfume, a cold agent or a limiting agent. The amount of the additional component such as the above-mentioned moisturizing agent can be easily selected by those skilled in the art within a range not to impair the purpose and effect of the present invention, and the amount thereof is 0.01 to 5% by weight, specifically 0.01 to 3% % ≪ / RTI >
상기 조성물은 조성물 총 중량에 대하여 0.001 중량% 내지 80 중량%, 예를 들면, 0.01 중량% 내지 60 중량%, 0.01 중량% 내지 40 중량%, 0.01 중량% 내지 30 중량%, 0.01 중량% 내지 20 중량%, 0.01 중량% 내지 10 중량%, 0.01 중량% 내지 5 중량%, 0.05 중량% 내지 60 중량%, 0.05 중량% 내지 40 중량%, 0.05 중량% 내지 30 중량%, 0.05 중량% 내지 20 중량%, 0.05 중량% 내지 10 중량%, 0.05 중량% 내지 5 중량%, 0.1 중량% 내지 60 중량%, 0.1 중량% 내지 40 중량%, 0.1 중량% 내지 30 중량%, 0.1 중량% 내지 20 중량%, 0.1 중량% 내지 10 중량%, 또는 0.1 중량% 내지 5 중량%의 락토바실러스 람노서스 JC1225 균체를 포함할 수 있다.The composition may be present in an amount of from 0.001% to 80%, such as from 0.01% to 60%, from 0.01% to 40%, from 0.01% to 30%, from 0.01% to 20% %, 0.01% to 10%, 0.01% to 5%, 0.05% to 60%, 0.05% to 40%, 0.05% to 30%, 0.05% to 20% From 0.05% to 10%, from 0.05% to 5%, from 0.1% to 60%, from 0.1% to 40%, from 0.1% to 30%, from 0.1% to 20% % To 10% by weight, or 0.1% to 5% by weight of Lactobacillus laminosus JC1225.
다른 양상은 락토바실러스 람노서스 JC1225 균주를 유효성분으로 포함하는 피부 손상을 개선 또는 예방하기 위한 건강기능식품용 조성물을 제공한다.Another aspect provides a composition for a health functional food for improving or preventing skin damage comprising Lactobacillus lambosus JC1225 strain as an active ingredient.
상기 건강기능식품용 조성물은 당해 기술분야에 공지되어 있는 통상적인 건강기능식품의 제형으로 제제화될 수 있다.The health functional food composition may be formulated into a conventional health functional food formulation known in the art.
상기 건강기능식품은 예를 들어 산제, 과립제, 정제, 환제, 캅셀제, 현탁액, 유제, 시럽제, 침제, 액제, 엑스제 등의 일반적인 제형으로 제조될 수도 있고, 육류, 소세지, 빵, 쵸코렛, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 젤리, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알콜 음료 및 비타민 복합제 등의 임의의 건강식품 형태로 제조될 수도 있다. 상기 건강식품의 제제화를 위해 식품학적으로 허용 가능한 담체 또는 첨가제를 사용할 수 있으며, 제조하고자 하는 제형의 제조에 당해 기술분야에서 사용 가능한 것으로 공지되어 있는 임의의 담체 또는 첨가제가 이용될 수 있다. 상기 첨가제로서 각종 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있다. 그 이외에도 천연 과일쥬스, 과일쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 첨가제 성분은 독립적으로 또는 조합하여 사용할 수 있으며, 첨가제의 비율은 크게 중요하진 않지만 본 발명의 조성물 100 중량부 당 0.01 ~ 0.1 중량부의 범위에서 선택될 수 있다. The health functional food may be prepared by conventional formulations such as powders, granules, tablets, pills, capsules, suspensions, emulsions, syrups, Beverages, tea, drinks, alcoholic beverages, and vitamins, such as, for example, beverages, snacks, confectionery, pizza, ramen, other noodles, gums, jellies, ice creams, A pharmaceutically acceptable carrier or additive may be used for the formulation of the health food, and any carrier or additive known to be usable in the art for the preparation of the formulation to be prepared may be used. As the above-mentioned additives, there can be used various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, Carbonating agents, and the like. In addition, it may contain natural fruit juice, fruit juice beverage and flesh for the production of vegetable beverages. These additive components may be used independently or in combination, and the proportion of the additive is not critical, but may be selected in the range of 0.01 to 0.1 part by weight per 100 parts by weight of the composition of the present invention.
상기 건강기능식품용 조성물 중의 상기 락토바실러스 람노서스 JC1225 균체의 함량은 사용 목적(예방 또는 개선)에 따라 적합하게 결정될 수 있다. 일반적으로, 전체 식품 중량의 0.01 내지 15 중량%로 포함할 수 있으며, 음료로서 제조될 경우 100 mL를 기준으로 0.02 내지 10 g의 비율로 함유할 수 있다. The content of the Lactobacillus laminosus JC1225 cells in the composition for health functional food may be suitably determined according to the intended use (prevention or improvement). Generally, it may contain 0.01 to 15% by weight of the total food weight, and when it is prepared as a beverage, it may contain 0.02 to 10 g based on 100 mL.
상기 음료는 상기 락토바실러스 람노서스 JC1225 균체 이외의 다른 성분을 더 포함할 수 있으며, 통상적으로 음료에 사용되는 다양한 향미제 또는 천연 탄수화물 등을 더 함유할 수 있다. 상기 천연 탄수화물로는 단당류(예: 포도당, 과당 등), 이당류(예: 말토즈, 수크로즈 등), 다당류(예: 덱스트린, 시클로덱스트린 등)와 같은 통상적인 당 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이 함유될 수 있다. 또한, 향미제로서 천연 향미제(예: 타우마틴, 스테비아 추출물 등) 및 합성 향미제(예: 사카린, 아스파탐 등)를 함유할 수 있다. 상기 천연 탄수화물의 비율은 음료 100 mL 당 일반적으로 약 1 내지 20g, 바람직하게는 약 5 내지 12g으로 함유될 수 있다.The beverage may further contain components other than the Lactobacillus lambsosus JC1225 cells, and may further contain various flavors or natural carbohydrates commonly used in beverages. Examples of the natural carbohydrate include conventional sugars such as monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; and sugars such as xylitol, sorbitol, erythritol, Of sugar alcohols may be contained. In addition, as a flavoring agent, a natural flavoring agent (e.g., tau Martin, stevia extract, etc.) and a synthetic flavoring agent (e.g., saccharin, aspartame, etc.) may be contained. The ratio of the natural carbohydrate may be generally about 1 to 20 g, preferably about 5 to 12 g per 100 mL of beverage.
상기 설명에서 이미 언급된 용어 또는 요소에 대해서는 따로 언급하는 것이 없는한 이미 상술한 바와 같다.The terms or elements already mentioned in the above description are already described above unless otherwise stated.
락토바실러스 람노서스 JC1225 균주 및 이의 사균체는 다른 유산균주와는 차별화되는 피부 보습, 주름 개선, 및 피부 미백 효과를 가지고 있어, 피부 손상을 개선하기 위한 조성물 등에 유용하게 사용될 수 있다.The Lactobacillus lambsus JC1225 strain and its dead cells have skin moisturizing, wrinkle and skin whitening effects different from those of other lactic acid bacteria, and thus can be effectively used for compositions for improving skin damage.
도면 중 *은 대조군에 비해 유의성이 있는 것을 나타내며(P<0.0001); #은 자외선 조사군에 비해 유의성이 있는 것을 나타내고(P<0.0001); CON은 대조군; UV는 자외선 조사군; TLC는 사멸화 락토바실러스 카세이 처리군; TLA는 사멸화 락토바실러스 아시도필루스 처리군; TLB는 사멸화 락토바실러스 브레비스 처리군; 및 TLR은 사멸화 락토바실러스 람노서스의 처리군을 의미한다.
도 1은 사멸화 락토바실러스 람노서스의 경피수분 손실량 억제효능을 확인한 결과이다.
도 2는 사멸화 락토바실러스 람노서스의 피부수분량을 평가한 결과이다.
도 3은 사멸화 락토바실러스 람노서스의 프로콜라겐량 측정을 통한 주름 개선 효능을 평가한 결과이다.
도 4는 사멸화 락토바실러스 람노서스의 MMP-1량 측정을 통한 주름 개선 효능을 평가한 결과이다.
도 5는 사멸화 락토바실러스 람노서스의 멜라닌 생성 억제율을 평가한 결과이다.
도 6은 사멸화 락토바실러스 람노서스의 피부 미백 효과를 측정한 결과이다.
도 7은 사멸화 락토바실러스 람노서스를 투여한 환자의 두피 상태가 개선되는 모습을 나타내는 사진이다.
도 8은 사멸화 락토바실러스 람노서스를 투여한 환자의 등 피부 상태가 개선되는 모습을 나타내는 사진이다.* In the figure indicates that there is a significant difference (P < 0.0001) compared to the control group; # Was significantly (P <0.0001) higher than ultraviolet irradiation group; CON is the control; UV is an ultraviolet irradiation group; TLC was treated with a peroxidized Lactobacillus casein; TLA was treated with killed Lactobacillus acidophilus; TLB was treated with killed Lactobacillus brevis; And TLR refers to the treated group of the killed Lactobacillus lambosus.
Fig. 1 shows the results of confirming the effect of inhibiting the transdermal water loss of the peroxidized Lactobacillus lambosus.
Fig. 2 shows the result of evaluating the skin moisture content of the peroxidized Lactobacillus lambosus.
FIG. 3 shows the results of evaluating the wrinkle-reducing effect by measuring the amount of procollagen in the peroxidized Lactobacillus lambosus.
FIG. 4 shows the results of evaluating the wrinkle-reducing effect of MMP-1 in the killed lipobacterium laminocus.
Fig. 5 shows the results of evaluating the rate of inhibition of melanin formation in the peroxidized Lactobacillus lambosus.
Fig. 6 shows the result of measurement of the skin whitening effect of the peroxidized Lactobacillus lambosus.
FIG. 7 is a photograph showing a state in which the scalp condition of a patient administered with an endosulfanylated Lactobacillus lambosus is improved.
FIG. 8 is a photograph showing a state in which the back skin condition of a patient administered with the killed lipobactor lambsosus is improved.
이하 본 발명을 실시예를 통하여 보다 상세하게 설명한다. 그러나, 이들 실시예는 본 발명을 예시적으로 설명하기 위한 것으로 본 발명의 범위가 이들 실시예에 한정되는 것은 아니다.Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.
<< 비교예Comparative Example 1> 1> 사멸화Extinction 락토바실러스Lactobacillus 카세이Kasei 배양액 건조물 제조 Manufacture of culture dried material
MRS 액체배지에서 37℃에서 24시간 배양 후 여과한 락토바실러스 카세이(Lactobacillus casei)를 120℃에서 30분동안 열사하여 사멸화하고, 20℃에서 냉각하여, 이를 동결건조하였다. 사멸화 락도바실러스 카세이(tyndallized Lactobacillus casei: TLC) 건조물을 수득하였다. Lactobacillus casei , which had been filtered and cultured in an MRS liquid medium at 37 ° C for 24 hours, was thermally killed at 120 ° C for 30 minutes to be killed, cooled at 20 ° C and lyophilized. Drying of the tyndallized Lactobacillus casei (TLC) was also obtained.
<< 비교예Comparative Example 2> 2> 사멸화Extinction 락토바실러스Lactobacillus 아시도필루스Ashidopoulos 배양액 건조물 제조 Manufacture of culture dried material
MRS 액체배지에서 37℃에서 24시간 배양 후 여과한 락토바실러스 아시도필루스(Lactobacillus acidophilus)를 120℃에서 30분동안 열사하여 사멸화하고, 20℃에서 냉각하여, 이를 동결건조하였다. 사멸화 락도바실러스 아시도필루스(tyndallized Lactobacillus acidophilus: TLA) 건조물을 수득하였다. Lactobacillus acidophilus , which was filtered and cultured in an MRS liquid medium at 37 ° C for 24 hours, was thermally killed at 120 ° C for 30 minutes to be killed, cooled at 20 ° C, and lyophilized. A deadly lacquer also yielded the dried T. tyndallized Lactobacillus acidophilus (TLA).
<< 비교예Comparative Example 3> 3> 사멸화Extinction 락토바실러스Lactobacillus 브레비스Brevis 배양액 건조물 제조 Manufacture of culture dried material
MRS 액체배지에서 37℃에서 24시간 배양 후 여과한 락토바실러스 브레비스(Lactobacillus brevis)를 120℃에서 30분동안 열사하여 사멸화하고, 20℃에서 냉각하여, 이를 동결건조하였다. 사멸화 락도바실러스 브레비스(tyndallized Lactobacillus brevis: TLB) 건조물을 수득하였다. Lactobacillus brevis , which had been cultured in an MRS liquid medium at 37 ° C for 24 hours, was thermally killed at 120 ° C for 30 minutes to be killed, cooled at 20 ° C and lyophilized. A desiccation rock also yielded the dried T. tyndallized Lactobacillus brevis (TLB).
<< 실시예Example 1> 1> 사멸화Extinction 락토바실러스Lactobacillus 람노서스Lambrosse JC1225 스트레인 배양액 건조물 제조 Production of JC1225 strain culture
(1) 균주 동정(1) Identification of strain
건강한 성인 장내 미생물을 얻기 위해 채취된 분변 속의 균주를 동정하고자 0.85% 살린(saline) 용액 50 mL에 분변 1 g을 넣고, 스토마커(stomacher)(simple mixer, 3M)를 이용하여 3분간 교반하였다. 교반 후 고형물 제거를 위해 페이퍼 필터(paper filter)를 이용하여 여과 후 1 mL을 취하여 0.85% 살린 희석액으로 105까지 십진희석하였다. 원액, 10 ~ 105의 희석액을 0.2% BPB(bromophenol blue) 지시약 1%가 첨가된 MRS, LB, TSB 한천에 각각 농도별로 도말하여 30℃의 혐기배양기에서 48~72시간 배양하였다. 콜로니 주변이 노랗게 변한 균주를 육안으로 관찰하여 형태 및 색 등이 서로 다른 콜로니들을 각각 새로운 MRS 한천 플레이트에 선조접종(streaking)하여 37℃에서 48시간 배양하였다. 배양 후 얻은 단일 콜로니(single colony)는 2회 이상의 계대를 통해 순수배양을 유도하고, 얻은 콜로니는 16S rRNA 유전자의 서열분석을 통해 종을 확인하였다. 이들 중 서열번호 1의 16s rRNA 유전자를 갖는 락토바실러스 람노서스 JC1225 균주를 선별하였다.In order to obtain a healthy adult intestinal microorganism, 1 g of feces was added to 50 mL of a 0.85% saline solution and the mixture was stirred for 3 minutes using a stomacher (simple mixer, 3M). After stirring, 1 mL of the filtrate was removed using a paper filter to remove solids, and the solution was decanted to 10 5 with 0.85% saline. 10 to 10 5 dilutions were plated on MRS, LB, and TSB agar supplemented with 1% of 0.2% BPB (bromophenol blue) indicator and incubated for 48 to 72 hours in an anaerobic incubator at 30 ° C. Colonies with a yellow color around the colonies were visually observed, and colonies having different shapes and colors were each streaked onto new MRS agar plates and cultured at 37 DEG C for 48 hours. A single colony obtained after cultivation induced pure culture through two or more passages, and the obtained colonies were identified by sequencing the 16S rRNA gene. Of these, Lactobacillus lambosus JC1225 strain having the 16s rRNA gene of SEQ ID NO: 1 was selected.
(2) 배양 건조물 수득(2) Obtain culture culture
MRS 액체배지에서 37℃에서 24시간 배양한 후 락토바실러스 람노서스 JC1225(Lactobacillus rhamnosus JC1225) 균주 배양액을 120℃에서 30분동안 열사하여 사멸화 후 건조하여 20℃에서 냉각하여, 이를 동결 건조하였다. 사멸화 락도바실러스 람노서스(tyndallized Lactobacillus rhamnosus: TLR) 건조물을 수득하였다.After culturing in MRS liquid medium at 37 DEG C for 24 hours, Lactobacillus rhamnosus JC1225 JC1225) was incubated at 120 ° C for 30 minutes, and then killed and dried at 20 ° C and lyophilized. The anterolytic rock is also a tyndallized Lactobacillus rhamnosus : TLR).
<< 실시예Example 2> 2> 사멸화Extinction 락토바실러스Lactobacillus 람노서스Lambrosse JC1225 스트레인 JC1225 strain 처리군의Treatment group 보습효능평가 Evaluation of moisturizing efficacy
1. 실험동물 및 시료투여1. Experimental animal and sample administration
7주령 수컷 무모쥐(male HR-1, Hairless mice, Japan SLC Inc.)를 중앙실험동물로부터 구입하여 1주 동안 적응시킨 후 사용하였다. 적응기간 중 일반 상태를 관찰하여 건강한 상태의 동물을 시험에 사용하였다. 사육환경은 온도 23±3℃, 습도 50±5%, 명암주기 12시간(07:00-19:00/조명시간)으로 유지하였다. 시험기간 중 실험동물은 군 당 7마리로 사육하였고, 사료는 마우스 전용사료 5L79(Charles river, USA)를 자유로이 급이 하였으며, 음수는 자외선으로 소독한 상수도수를 자유로이 급이 하였다. 시료의 투여는 대조군(control), 자외선 처리군(UV), 사멸화 유산균 투여군(TLC, TLA, TLB, TLR)으로 총 6개 군으로 나누어 실험을 하였다. 시료투여는 마우스 존대를 이용하여 경구 투여를 실시하였다. 투여기간은 총 12주로 주 5일 동안 투여하였다.Seven-week-old male hairless mice (male HR-1, Hairless mice, Japan SLC Inc.) were purchased from the central laboratory animals and used for 1 week. Animals in healthy state were used for the test by observing the general condition during the adaptation period. The breeding environment was maintained at a temperature of 23 ± 3 ° C, a humidity of 50 ± 5%, and a light period of 12 hours (07: 00-19: 00 / lighting time). During the test period, the experimental animals were fed with 7 livers per group. Feeds were fed 5L79 (Charles river, USA) for mouse only and free water was added for drinking water. Tests were divided into six groups: control, ultraviolet (UV), and peroxidized lactic acid bacteria (TLC, TLA, TLB, TLR). Samples were administered orally using a mouse mouse. The administration period was 12 weeks for a total of 5 days.
2. 자외선 조사2. Ultraviolet irradiation
피부 손상을 유도하기 위하여, 대조군을 제외한 실험군에 12주 동안 주 3회 자외선 조사를 실시하였다. 자외선은 UVB 램프 (Mineralight UV Display lamp, UVP, USA)를 사용하였다. 자외선 조사량은 1 내지 7주간은 60 mJ/㎠, 8 내지 12주는 90 mJ/㎠으로 12주 동안 조사하였다. 자외선 조사량은 광측정기 (Delta OHM, Italy)를 이용하여 광량을 측정한 후 조사시간을 조절하였다.To induce skin damage, the experimental group except for the control group was irradiated three times a week for 12 weeks. The ultraviolet rays were UVB lamp (Mineralight UV Display lamp, UVP, USA). The ultraviolet irradiation dose was 60 mJ /
3. 3. 경피Percutaneous 수분 손실량( Water loss ( TEWLTEWL : : transepidermaltransepidermal waterwater lossloss ) 분석) analysis
보습 효과를 확인하기 위하여 각 실험군의 무모 생쥐의 등 피부에서 경피 수분 손실량 분석을 실시하였다. 경피 수분 손실량 수치는 피부로부터 발산되는 수분량으로서, 수치가 높을수록 피부의 보습 기능이 떨어져 있음을 의미하며, 피부 고유의 장벽 기능이 손상되었음을 나타낸다. 경피 수분 증발량은 항온, 항습 조건(23℃, 상대습도: 50%)에서 투와미터(Tewameter Courage & Khazaka, Germany)를 이용하여 피부에서 증발하는 수분의 양을 면적과 시간에 따라 계산하였으며, 증발하는 수분의 양(g/m/hr)을 보습 전자센서로 읽고, 이를 수치화하여 피부의 보습력을 측정하였다. 결과를 도 1에 나타내었다. 도 1과 같이, 자외선조사에 의해 피부 장벽이 손상되어 피부 보습 기능이 떨어졌음을 확인할 수 있었으며 (p<0.0001), 사멸화 락토바실러스 람노서스(TLR) 처리군에 의하여 자외선에 의해 유발되는 피부 수분 손실을 방지할 수 있음을 확인하였다 (p<0.0001). 반면에 사멸화 락토바실러스 아시도필루스(TLA)도 피부 수분 손실을 약하게 방지하는 것을 확인하였으나, 사멸화 락토바실러스 카세이(TLC), 사멸화 락토바실러스 브레비스(TLB)는 피부 수분 손실 방지효과가 나타나지 않았다. To confirm the moisturizing effect, transcutaneous water loss analysis was performed on the dorsal skin of reared mice in each experimental group. The amount of transdermal water loss is the amount of water emanating from the skin. The higher the value, the lower the moisturizing function of the skin, indicating that the inherent barrier function of the skin is impaired. The amount of water vapor evaporated from the skin was calculated by area and time using a Tewameter Courage & Khazaka, Germany at constant temperature and humidity conditions (23 ° C, 50% relative humidity) (G / m / hr) was read by a moisturizing electronic sensor, and the moisture content of the skin was measured. The results are shown in Fig. As shown in Fig. 1, it was confirmed that the skin barrier was damaged due to UV irradiation (p <0.0001), and skin moisture loss caused by ultraviolet rays (P < 0.0001). ≪ / RTI > On the other hand, it was confirmed that the peroxidized Lactobacillus acidophilus (TLA) also weakly prevented skin water loss, but the deadly Lactobacillus casei (TLC) and the killed Lactobacillus brevis (TLB) I did.
4. 피부수분량 측정4. Skin moisture measurement
각 실험군의 무모 생쥐의 등 피부에서 피부수분량 분석을 실시하였다. 무모 생쥐의 등에 사멸화 락토바실러스를 14% 농도로 처리한 후, 3시간 뒤 항온항습 조건 (23℃, 상대습도 50%)에서 코니오미터 (Corneometer Courage & Khazaka, Germany)를 이용하여 수분량을 측정하였다. 센서를 이용하여 수분의 이온정도를 측정하고, 피부의 표피 내에 존재하는 수분의 양을 수치화하여 수분의 양을 계산함으로써 보습력을 측정하였다.Skin moisture analysis was performed on the dorsal skin of reared mice of each experimental group. After 3 hours, the water content was measured using a corneometer (Corneometer Courage & Khazaka, Germany) under constant temperature and humidity conditions (23 ° C,
그 결과, 도 2에서와 같이 자외선 조사군은 피부수분량이 대조군에 비해서 감소하였음을 확인하였고(p<0.0001), 사멸화 락토바실러스 람노서스(TLR) 처리군은 자외선 조사군에 비해서 뚜렷하게 증가된 피부수분 함유량이 관찰되었다. 반면에 사멸화 락토바실러스 아시도필루스(TLA)가 처리된 실험군도 다소 피부 수분 함유량이 증가한 것을 확인하였으나, 사멸화 락토바실러스 카세이(TLC), 사멸화 락토바실러스 브레비스(TLB)는 피부수분 함유량이 높지 않았다. 이런 결과를 토대로 사멸화 락토바실러스 람노서스 처리군이 피부 보습효과가 있음을 확인하였다.As a result, as shown in FIG. 2, it was confirmed that the amount of skin moisture in the ultraviolet irradiated group was decreased compared to the control group (p <0.0001), and the epidermal Lactobacillus lambosus (TLR) treated group had a significantly increased skin Moisture content was observed. On the other hand, in the treated group treated with the deadly lactobacillus acidophilus (TLA), the skin moisture content was somewhat increased. However, the peroxidized Lactobacillus casei (TLC) and the killed Lactobacillus brevis (TLB) It was not high. Based on these results, it was confirmed that the skin-moisturizing effect of the peroxidized Lactobacillus lambosus treatment group was confirmed.
<< 실시예Example 3> 3> 사멸화Extinction 락토바실러스Lactobacillus 람노서스Lambrosse JC1225 스트레인의 주름개선 효능평가 Evaluation of JC1225 strain wrinkle-reducing efficacy
1. One. TLR이TLR 처리된 세포 내 콜라겐 생성량 분석 Analysis of collagen production in treated cells
인간섬유아세포(Human fibroblast) 유래의 Hs68(ATCC; CRL-1635) 세포를 1.5 x 105 세포/웰으로 6-웰 플레이트에 분주하고 세포가 바닥 면에 부착되도록 24시간 배양하였다. Hs68 세포가 바닥 면에 부착한 후에 배양액을 제거하고, 세포독성시험 결과를 토대로 세포 생존율이 100%인 농도부터 TLA, TLC, TLB 및 TLR을 각각 8배 희석하여 배지에 처리하고 24시간 동안 배양하였다. 24시간 뒤, 배양액과 세포를 회수하고 세포 용해물로 단백질을 정량 한 뒤 ELISA 키트(mdbioproducts; Cat. M036007)를 사용하여 배양액에 생성된 콜라겐의 양을 측정하였다. Human fibroblasts (Human fibroblast) origin of Hs68 (ATCC; CRL-1635) were cultured 24 hours the cells 1.5 x 10 5 cells / well in 6-well plates in cell division and to be attached to the bottom surface. After adherence of Hs68 cells to the bottom surface, the culture medium was removed and the medium was diluted 8 times with TLA, TLC, TLB and TLR starting from the cell viability of 100% based on the cytotoxicity test and cultured for 24 hours . After 24 hours, the culture medium and cells were recovered and protein was quantified with a cell lysate, and the amount of collagen produced in the culture was measured using an ELISA kit (mdbioproducts; Cat. M036007).
도 3은 0.00004 wt%(사균체양/Hs68 배양배지)로 시험물질을 Hs68에 처리한 결과를 나타낸다. 사멸화 락토바실러스 람노서스(TLR)가 처리된 Hs68 세포는 세포 내에서 콜라겐 생성이 증가되는 효과가 있음이 확인되었다. 반면에 사멸화 락토바실러스 아시도필루스(TLA), 사멸화 락토바실러스 카세이(TLC), 사멸화 락토바실러스 브레비스(TLB)는 세포내 콜라겐 생성효과가 높지 않았다. Fig. 3 shows the results of treatment of the test substance with Hs68 at 0.00004 wt% (amount of dead cells / Hs68 culture medium). It was confirmed that Hs68 cells treated with killed Lactobacillus lansosus (TLR) had an effect of increasing collagen production in the cells. On the other hand, intracellular collagen production effect was not high in the deadly Lactobacillus acidophilus (TLA), the killed Lactobacillus casei (TLC), and the killed Lactobacillus brevis (TLB).
2. 2. TLR이TLR 처리된 세포 내 Within treated cells MMPMMP (( humanhuman matrixmatrix metalloproteinasemetalloproteinase )-1 생성량 분석) -1 Production amount analysis
인간섬유아세포(Human fibroblast) 유래의 Hs68(ATCC; CRL-1635) 세포를 1.5 x 105 세포/웰로 6-웰 플레이트에 분주하고 세포가 바닥 면에 부착되도록 24시간 배양하였다. 세포가 바닥 면에 부착한 후에 배양액을 제거하고 PBS로 2회 세척한 후 PBS에서 UVB(80 mJ/cm2; 312 nm)조사를 하였다. 세포독성시험 결과를 토대로 세포 생존율이 100%인 농도부터 2~16배 희석한 배지를 처리하고 48시간 동안 배양하였다. 48시간 뒤, 배양액과 세포를 회수하고 세포 용해물로 단백질을 정량 한 뒤 ELISA 키트(Amersham; Cat. RPN2610)를 사용하여 배양액에 함유된 MMP-1양을 측정하였다.Human fibroblast cells (Human fibroblast) origin of Hs68 (ATCC; CRL-1635) were cultured 24 hours dividing the cell to 1.5 x 10 5 cells / well and 6-well plate so the cells are attached to the bottom surface. After the cells adhered to the bottom, the culture solution was removed, and the cells were washed twice with PBS, followed by UVB (80 mJ / cm 2 ; 312 nm) in PBS. Based on the results of the cytotoxicity test, the medium was diluted 2 to 16 times with a cell viability of 100%, and cultured for 48 hours. After 48 hours, the culture medium and cells were recovered and protein was quantified with a cell lysate, and the amount of MMP-1 contained in the culture solution was measured using an ELISA kit (Amersham; Cat. RPN2610).
도 4는 0.0025 wt%(사균체양/Hs68 배양배지)로 시험물질을 Hs68에 처리한 결과를 나타낸다. 사멸화 락토바실러스 람노서스(TLR) 처리군은 세포내에서 MMP1 생성을 억제하는 것이 확인되었다. 반면에 사멸화 락토바실러스 아시도필루스(TLA), 사멸화 락토바실러스 카세이(TLC), 사멸화 락토바실러스 브레비스(TLB)는 MMP-1 억제효과가 나타나지 않았다. 이런 결과를 토대로 사멸화 락토바실러스 람노서스 처리군이 주름개선효과가 있음을 확인하였다.Figure 4 shows the results of treatment of the test material with Hs68 at 0.0025 wt% (amount of dead cells / Hs68 culture medium). It was confirmed that the treatment with the killed Lactobacillus lambosus (TLR) inhibited the production of MMP1 in the cells. On the other hand, there was no MMP-1 inhibitory effect of the killed lipase Bacillus acidophilus (TLA), the killed lipase Bacillus casei (TLC), and the killed lipase Bacillus brevis (TLB). Based on these results, it was confirmed that waxy laxity was improved in the group treated with peroxidized Lactobacillus lambosus.
<< 실시예Example 4> 4> 사멸화Extinction 락토바실러스Lactobacillus 람노서스Lambrosse JC1225 스트레인의 미백 효능 평가 Evaluation of whitening efficacy of JC1225 strain
1. 쥐의 색소 세포를 이용한 멜라닌 생성 억제효과 측정1. Measurement of inhibitory effect on melanin production by mouse pigment cells
C57BL/6 마우스 유래의 쥐의 색소세포(Mel-Ab cell)를 DMEM(Dulbecco’s modified Eagle’s media)에 10% 우태반 혈청, 100nM 2-O-테트라데카노일포르빌 (tetradecanoyphorbil)-13-아테이트, 1nM 콜레라 독소(choleratoxin)을 첨가한 배지에서 37℃, 5% CO2의 조건에서 배양하였다. 배양된 Mel-Ab 세포를 0.25% 트립신-EDTA로 떼어낸 다음, 24-웰 플레이트에 105 세포/웰(cells/well)의 농도로 세포를 배양하고 이틀째부터 3일 연속으로 10 ppm의 시험물질을 가하여 배양하였다. TLA, TLB, TLC, 및 TLR에 대한 멜라닌 생성 억제 효과를 비교하기 위하여 대조군으로 코지산을 사용하였다. 배양액을 제거하고, PBS로 세척한 후, 1 N 수산화나트륨으로 세포를 녹여 400 nm에서 흡광도를 측정한 후, 하기 수학식 1에 따라 멜라닌 생성 억제율을 계산하여 그 결과는 도 5에 나타내었다(Dooley의 방법). Mouse C57BL / 6 mouse mel-Ab cells were inoculated into DMEM (Dulbecco's modified Eagle's media) supplemented with 10% fetal calf serum, 100 nM 2-O-tetradecanoyphorbil-13- The cells were cultured in a medium supplemented with 1 nM choleratoxin at 37 ° C and 5
<수학식 1>&Quot; (1) "
도 5와 같이, 사멸화 락토바실러스 람노서스(TLR)을 사용할 경우 35.8%의 멜라닌 생성 억제효과를 나타내어 미백효과가 매우 우수하였으며, 특히 미백 효과가 우수하다고 알려진 코지산에 비해서도 멜라닌 생성 억제효과가 우수함을 확인할 수 있었다. 반면에 사멸화 락토바실러스 카세이(TLC), 사멸화 락토바실러스 아시도필루스(TLA), 및 사멸화 락토바실러스 브레비스(TLB)는 멜라닌 생성 억제효과를 뚜렷하게 나타내지 않았다. As shown in FIG. 5, when using the killed lipobacillus rhamnose (TLR), the melanin production inhibitory effect was 35.8% and the whitening effect was excellent. Especially, the melanin production inhibitory effect was superior to that of kojic acid, . On the other hand, the peroxidized Lactobacillus casei (TLC), the killed Lactobacillus acidophilus (TLA), and the killed Lactobacillus brevis (TLB) did not show any significant inhibitory effect on melanin production.
2. 인체 2. Human body 피부에 대한 미백 효과Whitening effect on skin 실험 Experiment
건강한 12명의 남자를 대상으로 피검자의 상박 부위에 직경 1.5㎝의 구멍이 뚫린 불투명 테이프를 부착한 뒤, 각 피검자의 최소 홍반량(Minimal Erythema Dose))의 1.5~2배 정도의 자외선(UVB)을 조사하여 피부의 흑화를 유도하였다. 자외선 조사 후 시험물질이 각 1 중량% 포함되어 있는 용액(용매는 1,3-부틸렌그리콜:에탄올 = 7:3)을 흑화 부위에 바르고, 대조군으로는 코지산 3 중량% 및 용매(vehicle)만을 사용한 것과, 한 곳은 아무 것도 바르지 않은 것을 이용하였다. 이후 10주 동안 상태변화를 관찰하며 1주 단위로 피부의 색깔을 색차계 CR2002(일본, 미놀타 사)로 측정하였다. 도포 시작시점과 완료시점에서의 피부색의 차이(△L*)를 아래 수학식 2에 따라 계산하고, 이를 도 6에 나타내었다. 미백효과는 시료 도포 부위와 대조군 부위의 △L*의 비교로 판정하는데, △L*값이 2 정도일 경우는 침착된 색소의 미백화가 뚜렷한 경우이고, 1.5정도 이상이면 미백효과가 있다고 판정할 수 있다. Twelve healthy men were exposed to 1.5 to 2 times the UVB of the subject's minimum erythema dose after attaching an opaque tape with a hole of 1.5 cm in diameter at the upper abdomen of the subject. To induce blackening of the skin. After irradiation with ultraviolet rays, a solution containing 1 wt% of each test substance (1,3-butyleneglycol: ethanol = 7: 3 as a solvent) was applied to the blackening region. As a control group, 3 wt% And one where nothing was applied. The color of the skin was measured with a colorimeter CR2002 (Minolta, Japan) every 1 week by observing the state change for 10 weeks. The difference (? L *) of the skin color at the application start time and the completion time is calculated according to the following equation (2), which is shown in FIG. The whitening effect is judged by comparison of DELTA L * between the sample application region and the control region. When the DELTA L * value is about 2, the whitening of the deposited pigment is noticeable. When the DELTA L * value is about 1.5 or more, .
<수학식 2>&Quot; (2) "
ΔL* = 도포 완료 시점에서의 L* 값 - 도포 개시 시점에서의 L* 값DELTA L * = L * value at the time of completion of application - L * value at the start of application
도 6과 같이, 사멸화 락토바실러스 람노서스(TLR)을 사용할 경우 △L*값이 1.6 이상으로 미백 효과가 우수하였으며, 특히 미백 효과가 우수하다고 알려진 코지산에 비해서도 미백 효과가 우수함을 확인할 수 있었다. 반면에 사멸화 락토바실러스 카세이(TLC), 사멸화 락토바실러스 아시도필루스(TLA), 사멸화 락토바실러스 브레비스(TLB)에서는 미백효과가 뚜렷하게 나타나지 않았다. As shown in FIG. 6, when the saccharified lactobacillus lanxosus (TLR) was used, the ΔL * value was 1.6 or more, and the whitening effect was excellent. In particular, the whitening effect was superior to that of kojisan . On the other hand, whitening effect was not apparent in the peroxidized Lactobacillus casei (TLC), the killed Lactobacillus acidophilus (TLA), and the killed Lactobacillus brevis (TLB).
<< 실시예Example 5> 5> 사멸화Extinction 락토바실러스Lactobacillus 람노서스Lambrosse JC1225 스트레인의 염증 치료 효능 임상 평가 Clinical evaluation of JC1225 strain inflammation treatment efficacy
증례 1) 윤00(여/35세) Case 1) Yoon 00 (female / 35 years old)
상기인은 두피에 염증을 동반한 지루성 피부염이라는 진단을 병원에서 받고 별다른 호전 없는 상태로 두피 염증이 오히려 2년 동안 더 지속적으로 악화되고 탈모까지 진행되었다. 일 실시예에 따른 사멸화 락토바실러스 람노서스 JC1225 스트레인을 1일 3g씩 1회~2회씩 섭취하고 60일 만에 두피의 염증이 사라졌다. 아울러 탈모로 빠졌던 머리도 다시 나고 정상으로 돌아왔다. 도 7은 본 증례의 두피 치료 전후 사진을 나타낸다. The diagnosis of seborrheic dermatitis accompanied by inflammation in the scalp was received at the hospital, and the scalp inflammation continued to deteriorate for 2 years and continued to be alopecia. The intracutaneous Lactobacillus JC1225 strain according to one embodiment was ingested once or twice every 3 g of the day, and inflammation of the scalp disappeared within 60 days. In addition, the hair that had fallen into hair loss came back to normal. Fig. 7 shows photographs before and after the scalp treatment of the present case.
증례 2) 성00(여/50세)Case 2) Sex 00 (female / 50 years old)
상기인은 피부가 건조해서 하얗게 각질이 쌓이고 떨어지며 두피에 붉은 염증이 상존하는 상태에서 병원 피부과를 지속적으로 다녔지만 별다른 효과를 보지 못했다. 상태가 심해서 일 실시예에 따른 사멸화 락토바실러스 람노서스 JC1225 스트레인을 각 오전 3g, 오후 3g 섭취하게 하였고 약 60일 만에 피부에 하얀 각질이 없어지고 두피 염증도 없어졌다.The above-mentioned skin was dry, whiteness was piled up and fell off, and the scalp had red irritation. The condition was severe, so that the deadening Lactobacillus lambosus JC1225 strain according to one embodiment was ingested 3 g each morning and 3 g in the afternoon. In about 60 days, the skin had no white keratin and the scalp inflammation disappeared.
증례 3) 강00(여/33세)Case 3) 00 (female / 33 years old)
상기인은 평소에 피부가 건조하고 자기 전에 몸에서 하얀 각질이 떨어지고 소양감을 느꼈으며, 얼굴에도 잡티가 많았다. 일 실시예에 따른 사멸화 락토바실러스 람노서스 JC1225 스트레인을 하루 3g씩 1회씩 섭취하였다. 1개월 후부터 가려움이 현저히 없어지고 3개월 후에는 피부에 각질도 많이 개선이 되었다.The skin of the above-mentioned person was dry, and the white horny body fell from his body before he was sleeping, felt a sense of well-being, and had many dirt on his face. One gram of the peroxidized Lactobacillus laminosus JC1225 strain according to one embodiment was taken once per day. After 1 month, the itching was remarkably disappeared, and after 3 months, the skin was improved much.
증례 4) 정00(여/69세)Case 4) 00 (female / 69)
상기인은 얼굴과 등뒤에 붉은 염증이 퍼져있고 일부 염증부위에서는 피가 나고 소양감도 심했다. 일 실시예에 따른 사멸화 락토바실러스 람노서스 JC1225 스트레인을 각 3g씩 1일 1~2회 섭취했는데 약 15일부터 등의 염증이 가라앉기 시작해서 30일째에는 염증은 많이 호전되어 붉은 부위가 조금 남아 있는 상태로 호전되었다. 도 8은 본 증례의 등 염증 치료 전후 사진을 나타낸다.The person has red irritation behind the face and back, bleeding in some areas of inflammation, and feeling of pruritus. The ingestion of Lactobacillus rhamnosus JC1225 strain according to one embodiment was taken 1 to 2 times a day for each 3 g of the strain. After about 15 days, inflammation of the back began to sink, and on the 30th day, the inflammation improved a little, . 8 shows a photograph before and after the treatment of isoniatic inflammation in this case.
증례 5) 차0(남/33세) Case 5) Car 0 (male / 33 years old)
상기인은 입원했던 병원에서 total IgE가 1100 IU/ml로 측정되었다. 얼굴부위와 다리가 부위가 특히 염증과 짓물로 특히 심한 상태였다. 일 실시예에 따른 사멸화 락토바실러스 람노서스 JC1225 스트레인을 복용하고서 점차 좋아져 15일 후에 퇴원하고 2개월 만에 total IgE가 400 IU/ml로 측정되었고 지속적으로 호전이 되었다.Total IgE was measured at 1100 IU / ml in the hospital where the patient was hospitalized. The facial region and the leg region were especially severely affected by inflammation and debris. After taking 15 days of the saponin-treated Lactobacillus JC1225 strain according to one embodiment, the patient was discharged after 15 days, and the total IgE was measured at 400 IU / ml in 2 months, and it was continuously improved.
증례 6) 황00(남/45세) Case 6) Sulfur 00 (male / 45 years old)
상기인은 술을 많이 마시고 항상 얼굴이 상기되어 있고 피부가 붉고 건조한 상태였다. 일 실시예에 따른 사멸화 락토바실러스 람노서스 JC1225 스트레인을 하루 3g씩 섭취하였다. 처음에는 설사를 하였으나, 삼일 동안 섭취하지 않고 사일째부터는 섭취량을 1/3로 줄여 섭취했는데 설사 하지 않았다. 십일 후에는 다시 조금씩 먹는 양을 늘려 1일 3g씩 섭취했으며, 30일 후에는 얼굴의 붉은 색이 많이 좋아졌고 건조함이 많이 호전되었다.She had a lot of alcohol and always reminiscent of her face, and her skin was red and dry. Three grams of the peroxidized Lactobacillus lambosus JC1225 strain according to one embodiment was ingested. Initially, diarrhea, but did not eat for three days from the first day of the intake was reduced by one third of the diarrhea did not. After 10 days, the amount of eating a little bit more 3g a day, and 30 days later, the red color of the face was much better and dryness improved a lot.
증례 7) 김00(남/5세) Case 7) Kim 00 (male / 5 years old)
5세 남자 아이로서 밤에 피부가 건조하고 가려워서 잠을 거의 자지 못하고, 온몸? 긁어서 붉은 상처들이 나 있는 상태였다. 상태가 심해서 일 실시예에 따른 사멸화 락토바실러스 람노서스 JC1225 스트레인 조성물을 하루 5g씩 섭취케 하였고, 열흘 후에 많이 좋아지기 시작하여 한 달 후에는 아이가 밤에 잠을 잘 잘 수 있게 되고, 가려움이 거의 없어졌다.As a 5-year-old boy, his skin is dry and itchy at night. I was scratched and there were red wounds. The condition was so severe that the ingested Lactobacillus lambosus JC1225 strain composition according to one embodiment was ingested 5g / day, and after 10 days, it started to improve much so that the child could sleep well at night after a month, Almost lost.
증례 8) 김00(남/17세)Case 8) Kim 00 (male / 17 years old)
얼굴에 하얀 각질이 심하였고 팔다리 부위도 피부가 갈라지고 하얗게 각질이 버섯처럼 피어 있었다. 피부염까지 증가하여 본인이 괴로움을 호소하였다. 일 실시예에 따른 사멸화 락토바실러스 람노서스 JC1225 스트레인 조성물을 하루 3g씩 1일 1-2회씩 섭취케하였고, 25일 후쯤부터 많이 호전된 것이 관찰되고 60일 후에는 정상적인 생활이 가능하다고 하였다.The face was white keratinized, and the skin of the limbs was divided and the keratin was bloomed like white mushroom. My skin has been increasing, and I have complained of pain. One day, 3 g / day of the submerged Lactobacillus laminocyte JC1225 strain composition according to one embodiment was consumed 1-2 times a day. After 25 days, the improvement was observed and the normal life was possible after 60 days.
증례 9) 김00(여/24세) Case 9) Kim 00 (female / 24 years old)
취업준비 중에 발목과 무릎 뒤 접히는 부분과 손등 위에 아토피와 가려움증이 갑자기 심하게 나타났고, 일 실시예에 따른 사멸화 락토바실러스 람노서스 JC1225 스트레인 조성물을 하루 3g씩 1일 1-2회씩 20일 섭취 후 좋아지기 시작하여 60일 후 확연히 호전되었다.Atopic and itchy on the back of the ankle and the back of the knee suddenly appeared suddenly on the ankle and the back of the knee during preparation for work, and after ingestion of the deadly Lactobacillus lambosus JC1225 strain composition according to one embodiment for 20 days After 60 days from the start, it was obviously improved.
증례 10) 정00(남/49세)Case 10) 00 (male / 49 years old)
허리 둘레와 복부에 아토피성 피부염으로 고생하는 남성으로, 일 실시예에 따른 사멸화 락토바실러스 람노서스 JC1225 스트레인 조성물을 하루 3g씩 1일 1-2회씩 약30일 가량 섭취하였다. 섭취 후 40일 후에는 가려움과 붉은 반점이 없어지고 많이 호전되었다.A male suffering from atopic dermatitis in the waist and abdomen, ingested saccharide Lactobacillus rhamnosus JC1225 strain composition according to one embodiment about 3 times a day for 1 day 1-2 times for about 30 days. After 40 days of ingestion, itching and red spot disappearing and improved.
[수탁기관][Agency]
기탁기관명 : 한국생명공학연구원Institution name: Korea Biotechnology Research Institute
수탁번호 : KCTC13184BPAccession number: KCTC13184BP
수탁일자 : 20170111Checked on: 20170111
<110> CHUNG, Yong Hee <120> Composition for treatment of skin damage comprising tyndallized Lactobacillus rhamnosus JC1225 <130> PN112417 <160> 1 <170> KopatentIn 2.0 <210> 1 <211> 1852 <212> DNA <213> lactobacillus rhamnosus <400> 1 caccgccacg ccggcgcgga cggcgcgcga caccaccaaa tagcacaccg cccgccgcga 60 ccgactcacg acacacccag ccgacggaca ccatagagag cgcaccgcag cctctcgaca 120 cgcacggccg cgtgaagacg ccgcgcaccg cagagagtgc gaacaccgtg ccaccggaga 180 cacgcggacc gggccgaaca agaacacgga acgaaggaaa caggagaagc gtgagcgagt 240 gcaccgtagg tcagtaacga cggaaccagg accaccgtta gagttttgat caggctcagg 300 atgaacgctg gcggcgtgcc taatacatgc aagtcgaacg agttctgatt attgaaaggt 360 gcttgcatct tgatttaatt ttgaacgagt ggcggacggg tgagtaacac gtgggtaacc 420 tgcccttaag tgggggataa catttggaaa cagatgctaa taccgcataa atccaagaac 480 cgcatggttc ttggctgaaa gatggcgtaa gctatcgctt ttggatggac ccgcggcgta 540 ttagctagtt ggtgaggtaa cggctcacca aggcaatgat acgtagccga actgagaggt 600 tgatcggcca cattgggact gagacacggc ccaaactcct acgggaggca gcagtaggga 660 atcttccaca atggacgcaa gtctgatgga gcaacgccgc gtgagtgaag aaggctttcg 720 ggtcgtaaaa ctctgttgtt ggagaagaat ggtcggcaga gtaactgttg tcggcgtgac 780 ggtatccaac cagaaagcca cggctaacta cgtgccagca gccgcggtaa tacgtaggtg 840 gcaagcgtta tccggattta ttgggcgtaa agcgagcgca ggcggttttt taagtctgat 900 gtgaaagccc tcggcttaac cgaggaagtg catcggaaac tggaaaactt gagtgcagaa 960 gaaggacagt ggaactccat gtgtagcggt gaaatgcgta gatatatgga agaacaccag 1020 tggcgaaggc ggctgtctgg tctgtaactg acgctgaggc tcgaaagcat gggtagcgaa 1080 caggattaga taccctggta gtccatgccg taaacgatga atgctaggtg ttggagggtt 1140 tccgcccttc agtgccgcag ctaacgcatt aagcattccg cctggggagt acgaccgcaa 1200 ggttgaaact caaaggaatt gacgggggcc cgcacaagcg gtggagcatg tggtttaatt 1260 cgaagcaacg cgaagaacct taccaggtct tgacatcttt tgatcacctg agagatcagg 1320 tttccccttc gggggcaaaa tgacaggtgg tgcatggttg tcgtcagctc gtgtcgtgag 1380 atgttgggtt aagtcccgca acgagcgcaa cccttatgac tagttgccag catttagttg 1440 ggcactctag taagactgcc ggtgacaaac cggaggaagg tggggatgac gtcaaatcat 1500 catgcccctt atgacctggg ctacacacgt gctacaatgg atggtacaac gagttgcgag 1560 accgcgaggt caagctaatc tcttaaagcc attctcagtt cggactgtag gctgcaactc 1620 gcctacacga agtcggaatc gctagtaatc gcggatcagc acgccgcggt gaatacgttc 1680 ccgggccttg tacacaccgc ccgtcacacc atgagagttt gtaacacccg aagccggtgg 1740 cgtaaccctt ttagggagcg agccgtctaa ggtgggacaa atgattaggg tgaagtcgta 1800 caggggtaac ccgtaaaagt ggtaggattt tgttatttgt ttggttgtgg gg 1852 <110> CHUNG, Yong Hee <120> Composition for treatment of skin damage caused by tyndallized Lactobacillus rhamnosus JC1225 <130> PN112417 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 1852 <212> DNA <213> lactobacillus rhamnosus <400> 1 caccgccacg ccggcgcgga cggcgcgcga caccaccaaa tagcacaccg cccgccgcga 60 ccgactcacg acacacccag ccgacggaca ccatagagag cgcaccgcag cctctcgaca 120 cgcacggccg cgtgaagacg ccgcgcaccg cagagagtgc gaacaccgtg ccaccggaga 180 cacgcggacc gggccgaaca agaacacgga acgaaggaaa caggagaagc gtgagcgagt 240 gcaccgtagg tcagtaacga cggaaccagg accaccgtta gagttttgat caggctcagg 300 atgaacgctg gcggcgtgcc taatacatgc aagtcgaacg agttctgatt attgaaaggt 360 gcttgcatct tgatttaatt ttgaacgagt ggcggacggg tgagtaacac gtgggtaacc 420 tgcccttaag tgggggataa catttggaaa cagatgctaa taccgcataa atccaagaac 480 cgcatggttc ttggctgaaa gatggcgtaa gctatcgctt ttggatggac ccgcggcgta 540 ttagctagtt ggtgaggtaa cggctcacca aggcaatgat acgtagccga actgagaggt 600 tgatcggcca cattgggact gagacacggc ccaaactcct acgggaggca gcagtaggga 660 atcttccaca atggacgcaa gtctgatgga gcaacgccgc gtgagtgaag aaggctttcg 720 ggtcgtaaaa ctctgttgtt ggagaagaat ggtcggcaga gtaactgttg tcggcgtgac 780 ggtatccaac cagaaagcca cggctaacta cgtgccagca gccgcggtaa tacgtaggtg 840 gcaagcgtta tccggattta ttgggcgtaa agcgagcgca ggcggttttt taagtctgat 900 gtgaaagccc tcggcttaac cgaggaagtg catcggaaac tggaaaactt gagtgcagaa 960 gaaggacagt ggaactccat gtgtagcggt gaaatgcgta gatatatgga agaacaccag 1020 tggcgaaggc ggctgtctgg tctgtaactg acgctgaggc tcgaaagcat gggtagcgaa 1080 caggattaga taccctggta gtccatgccg taaacgatga atgctaggtg ttggagggtt 1140 tccgcccttc agtgccgcag ctaacgcatt aagcattccg cctggggagt acgaccgcaa 1200 ggttgaaact caaaggaatt gacgggggcc cgcacaagcg gtggagcatg tggtttaatt 1260 cgaagcaacg cgaagaacct taccaggtct tgacatcttt tgatcacctg agagatcagg 1320 tttccccttc gggggcaaaa tgacaggtgg tgcatggttg tcgtcagctc gtgtcgtgag 1380 atgttgggtt aagtcccgca acgagcgcaa cccttatgac tagttgccag catttagttg 1440 ggcactctag taagactgcc ggtgacaaac cggaggaagg tggggatgac gtcaaatcat 1500 catgcccctt atgacctggg ctacacacgt gctacaatgg atggtacaac gagttgcgag 1560 accgcgaggt caagctaatc tcttaaagcc attctcagtt cggactgtag gctgcaactc 1620 gcctacacga agtcggaatc gctagtaatc gcggatcagc acgccgcggt gaatacgttc 1680 ccgggccttg tacacaccgc ccgtcacacc atgagagttt gtaacacccg aagccggtgg 1740 cgtaaccctt ttagggagcg agccgtctaa ggtgggacaa atgattaggg tgaagtcgta 1800 caggggtaac ccgtaaaagt ggtaggattt tgttatttgt ttggttgtgg gg 1852
Claims (17)
상기 배양된 균주에 열을 가하여 사멸화하는 단계; 및
상기 사멸화된 균주를 동결 건조하는 단계를 포함하는 사멸화 균체를 제조하는 방법.Culturing Lactobacillus laminocus JC1225 strain;
Subjecting the cultured strain to heat to kill it; And
And lyophilizing the killed bacterial strain.
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| KR102074383B1 (en) * | 2019-04-26 | 2020-02-06 | 주식회사 락토메이슨 | Skin-whitening cosmetic composition comprising lactobacillus rhamnosus lm1011 having immunostimulating activity |
| KR102149102B1 (en) | 2019-07-05 | 2020-08-27 | 에스케이바이오랜드 주식회사 | Method of preparing dermobiotics block composition for skin hydration and skin regeneration |
| KR20210006687A (en) | 2019-07-09 | 2021-01-19 | 서울여자대학교 산학협력단 | Novel Paenisporosarcina quisquiliarum 17mud 1-1541 strain and its use |
| KR102296286B1 (en) * | 2020-06-16 | 2021-09-01 | 주식회사 바이오뱅크힐링 | Lactobacillus rhamnosus strain, and vesicles from thereof and anti-inflammation and anti-bacteria uses of thereof |
| WO2021242057A1 (en) * | 2020-05-28 | 2021-12-02 | 주식회사 바이오뱅크힐링 | Lactobacillus sp. strain, endoplasmic reticulum derived therefrom, and anti-inflammatory and antibacterial use thereof |
| KR20230112200A (en) * | 2022-01-19 | 2023-07-27 | 록야 주식회사 | Lactobacillus rhamnosus B3421 isolated from ginseng sprout and composition comprising the same |
| KR102582773B1 (en) * | 2022-07-28 | 2023-09-27 | 한국콜마홀딩스 주식회사 | Novel Lactobacillus sp. strain and skin condition improving uses of thereof |
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2017
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR102074383B1 (en) * | 2019-04-26 | 2020-02-06 | 주식회사 락토메이슨 | Skin-whitening cosmetic composition comprising lactobacillus rhamnosus lm1011 having immunostimulating activity |
| WO2020218888A1 (en) * | 2019-04-26 | 2020-10-29 | 주식회사 락토메이슨 | Cosmetic composition for skin whitening containing novel lactobacillus rhamnosus lm1011 as active ingredient |
| KR102149102B1 (en) | 2019-07-05 | 2020-08-27 | 에스케이바이오랜드 주식회사 | Method of preparing dermobiotics block composition for skin hydration and skin regeneration |
| KR20210006687A (en) | 2019-07-09 | 2021-01-19 | 서울여자대학교 산학협력단 | Novel Paenisporosarcina quisquiliarum 17mud 1-1541 strain and its use |
| WO2021242057A1 (en) * | 2020-05-28 | 2021-12-02 | 주식회사 바이오뱅크힐링 | Lactobacillus sp. strain, endoplasmic reticulum derived therefrom, and anti-inflammatory and antibacterial use thereof |
| KR102296286B1 (en) * | 2020-06-16 | 2021-09-01 | 주식회사 바이오뱅크힐링 | Lactobacillus rhamnosus strain, and vesicles from thereof and anti-inflammation and anti-bacteria uses of thereof |
| KR20230112200A (en) * | 2022-01-19 | 2023-07-27 | 록야 주식회사 | Lactobacillus rhamnosus B3421 isolated from ginseng sprout and composition comprising the same |
| KR102582773B1 (en) * | 2022-07-28 | 2023-09-27 | 한국콜마홀딩스 주식회사 | Novel Lactobacillus sp. strain and skin condition improving uses of thereof |
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