KR102495402B1 - Burkholderia vietnamiensis strain and its use for improving hair or scalp condition - Google Patents
Burkholderia vietnamiensis strain and its use for improving hair or scalp condition Download PDFInfo
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- KR102495402B1 KR102495402B1 KR1020200188265A KR20200188265A KR102495402B1 KR 102495402 B1 KR102495402 B1 KR 102495402B1 KR 1020200188265 A KR1020200188265 A KR 1020200188265A KR 20200188265 A KR20200188265 A KR 20200188265A KR 102495402 B1 KR102495402 B1 KR 102495402B1
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- South Korea
- Prior art keywords
- hair
- strain
- scalp
- alopecia
- culture medium
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Abstract
신규한 미생물, 그의 파쇄액, 배양액, 추출액, 발효액, 및 이들의 모발 또는 두피 상태 개선 용도에 관한 것이다. 일 양상에 따른 신규 버크홀데리아 비에트나미엔시스 균주에 의하면, 발모 촉진, 탈모 방지, 모근 강화, 두피 지질 억제, 비듬 방지, 및 두피 염증 억제 효과가 있으므로, 모발 또는 두피 상태의 개선, 예방, 또는 치료 용도로 다양하게 응용될 수 있다.It relates to novel microorganisms, their lysates, culture solutions, extracts, fermented solutions, and their use for improving hair or scalp conditions. According to the novel Burkholderia bietnamiensis strain according to one aspect, there are effects of promoting hair growth, preventing hair loss, strengthening hair roots, inhibiting scalp lipids, preventing dandruff, and inhibiting scalp inflammation, thereby improving, preventing, or treating hair or scalp conditions. It can be applied for various purposes.
Description
신규한 미생물, 그의 파쇄액, 배양액, 추출액, 발효액, 및 이들의 모발 또는 두피 상태 개선 용도에 관한 것이다. It relates to novel microorganisms, their lysates, culture solutions, extracts, fermented solutions, and their use for improving hair or scalp conditions.
인체에 있어서 모발은 두피에 있는 모낭에서 자라나며, 일생 동안 성장기(anagen), 퇴행기(catagen), 휴지기(telogen) 등의 주기가 지속적으로 반복되며 생성, 성장, 탈락을 거치게 된다(Paus et al., J Invest Dermatol. 113, pp.523-532, 1999). 모낭의 퇴행기에는 모낭의 성장이 중단되고, 세포사멸 및 세포외 기질의 재구성이 발생하게 되기 때문에, 모낭의 퇴행기 및 휴지기 상태의 모낭세포가 많아질수록 머리카락이 빠지고 다시 자라나지 않는 탈모 현상을 나타내게 된다.In the human body, hair grows from hair follicles on the scalp, and cycles of growth (anagen), catagen (catagen), and telogen (telogen) are continuously repeated throughout life and go through generation, growth, and shedding (Paus et al. , J Invest Dermatol. 113, pp.523-532, 1999). In the catagen phase of the hair follicle, the growth of the hair follicle is stopped, and apoptosis and reorganization of the extracellular matrix occur. Therefore, as the number of hair follicle cells in the catagen and telogen phases of the hair follicle increases, the hair falls out and does not grow back. .
모발은 눈에 보이는 모간부와 눈에 보이지 않는 모근부로 나누어지며, 모근부 중 모발이 자라는 뿌리를 모구(Hair bulb)라고 부르고, 그 아랫부분에 움푹 들어간 부분을 모유두(Hair papilla)라고 한다(도 1). 모유두는 모발의 발생에 있어 가장 중요한 부분으로, 혈관과 신경이 있어 영양분과 성장인자를 전달하여 모발의 성장을 촉진한다. 모유두세포(dermal papilla cell)의 사멸 및 생장 감소로 인해 모발의 수가 감소하기 때문에, 모발의 유지 및 증가를 위해서는 모유두세포의 증식이 필요하다.Hair is divided into the visible hair shaft and the invisible hair root, and among the hair roots, the root from which hair grows is called the hair bulb, and the recessed part at the bottom is called the hair papilla (Fig. One). The dermal papilla is the most important part in the development of hair, and has blood vessels and nerves to deliver nutrients and growth factors to promote hair growth. Since the number of hairs decreases due to the death and growth reduction of dermal papilla cells, proliferation of dermal papilla cells is required to maintain and increase hair.
또한, 두피 지질이 과다하게 분비가 되는 지루성 두피염의 경우, 모낭 피지선에서 분비되는 피지가 안드로겐 호르몬의 작용에 의해 증가되며, 피지가 증가할수록 두피에 존재하는 미생물의 양이 증가한다. 이런 두피 미생물은 두피의 노폐물을 발생시켜 가려움과 고통을 동반한 탈모 증상을 일으킬 수 있다(한국미용학회지, 제18권 제2호 (2012), pp. 279-289).In addition, in the case of seborrheic scalpitis in which scalp lipids are excessively secreted, sebum secreted from the sebaceous glands of the hair follicles is increased by the action of androgen hormones, and the amount of microorganisms present on the scalp increases as the sebum increases. These scalp microorganisms can generate wastes on the scalp and cause hair loss symptoms accompanied by itching and pain (Journal of the Korean Society of Aesthetics, Vol. 18, No. 2 (2012), pp. 279-289).
최근 환경 오염 및 도시 스트레스가 심각해지면서 탈모증을 가진 현대인이 많아짐에 따라 이를 개선할 수 있는 물질에 대한 관심이 높아지고 있다. 따라서, 모근부 중 모발 생성의 시작을 담당하는 모유두세포의 성장 인자 발현을 증가시켜 모발의 생장을 유도하여 모근을 강화하고 탈모 예방에 도움을 줄 수 있는 물질을 발명하고자 하였다. 또한, 두피 지질을 억제하는 미생물 균체 및 균주 배양액을 활용하여 피지에 의한 염증 및 가려움증을 해결하고, 나아가 탈모 증상을 예방하는 물질을 발명하고자 하였다.Recently, as environmental pollution and urban stress become more serious, as the number of modern people with alopecia increases, interest in materials capable of improving them is increasing. Therefore, an attempt was made to invent a substance capable of strengthening the hair root and helping to prevent hair loss by inducing hair growth by increasing the expression of growth factors in the dermal papilla cells responsible for the start of hair production in the hair root. In addition, it was attempted to invent a substance that solves sebum-induced inflammation and itching by utilizing microbial cells and strain cultures that inhibit scalp lipids, and further prevents hair loss symptoms.
이와 관련하여, 대한민국 등록특허공보 제10-2102578호는 다수의 화합물을 포함하는 발모 또는 육모 촉진용 화장료 조성물을 개시하고 있다. 그러나, 모발 또는 두피 상태를 개선할 수 있는 미생물에 대한 연구는 많지 않다.In this regard, Korean Patent Registration No. 10-2102578 discloses a cosmetic composition for promoting hair growth or hair growth containing a plurality of compounds. However, there are not many studies on microorganisms that can improve hair or scalp condition.
프로바이오틱스(Probiotics)는 인체에 유익한 작용을 하는 미생물을 총칭하는 말로 우리 몸에 유익(benefit)을 주는 미생물을 말한다. 현재까지 알려진 대부분의 프로바이오틱스는 유산균으로 알려져 있다. 프로바이오틱스는 인체에 여러 가지 유익작용을 통해 효과적인 효능이 발생되는 것으로 보고되었지만, 두피상재균과 두피의 상호관계에 대한 연구는 미비한 실정이다.Probiotics is a collective term for microorganisms that have a beneficial effect on the human body, and refers to microorganisms that benefit the body. Most probiotics known to date are known as lactic acid bacteria. Probiotics have been reported to produce effective effects through various beneficial effects on the human body, but studies on the interaction between scalp flora and scalp are insufficient.
이에, 두피 관련된 상태에 유용하게 사용될 수 있는 두피상재균의 개발이 필요하다. Therefore, it is necessary to develop a fungus on the scalp that can be usefully used for scalp-related conditions.
일 양상은 버크홀데리아 비에트나미엔시스(Burkholderia vietnamiensis) 균주를 제공하는 것이다. One aspect is to provide a Burkholderia vietnamiensis strain.
다른 양상은 상기 균주의 파쇄액, 배양액, 또는 배양액의 추출물을 제공하는 것이다. Another aspect is to provide a lysate of the strain, a culture medium, or an extract of the culture medium.
또 다른 양상은 버크홀데리아 비에트나미엔시스(Burkholderia vietnamiensis) 균주, 그의 파쇄액, 배양액, 또는 배양액의 추출물을 포함하는 조성물을 제공하는 것이다.Another aspect is to provide a composition comprising a Burkholderia vietnamiensis strain, a lysate thereof, a culture medium, or an extract of the culture medium.
또 다른 양상은 유효한 양의 상기한 조성물을 그를 필요로 하는 개체에 처리 또는 투여하는 단계를 포함하는 개체의 상태를 예방, 개선, 또는 치료하는 방법을 제공한다.Another aspect provides a method for preventing, ameliorating, or treating a condition in a subject comprising treating or administering to a subject in need thereof an effective amount of a composition described above.
일 양상은 버크홀데리아 비에트나미엔시스(Burkholderia vietnamiensis) 균주를 제공한다.One aspect provides a Burkholderia vietnamiensis strain.
상기 균주는 버크홀데리아 속(Burkholderia sp.)에 속하는 균주일 수 있다.The strain may be a strain belonging to the Burkholderia sp .
상기 균주는 서열번호 1과 약 95% 이상, 약 96% 이상, 약 97% 이상, 약 98% 이상, 약 99% 이상, 약 99.5% 이상, 또는 약 99.9% 이상의 서열 동일성을 갖는 16S rRNA를 포함하는 것일 수 있다. 상기 균주는 서열번호 1의 16S rRNA를 포함하는 것일 수 있다.The strain comprises a 16S rRNA having at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9% sequence identity with SEQ ID NO: 1 it may be The strain may contain the 16S rRNA of SEQ ID NO: 1.
본 명세서에서 사용된 용어 "서열 동일성(sequence identity)"은 특정 비교 영역에서 양 서열을 최대한 일치되도록 얼라인시킨 후 서열간의 아미노산 잔기 또는 염기의 동일한 정도를 의미한다. 서열 동일성은 특정 비교 영역에서 2개의 서열을 최적으로 얼라인하여 비교함으로써 측정되는 값으로서, 비교 영역 내에서 서열의 일부는 대조 서열(reference sequence)과 비교하여 부가, 삭제되어 있을 수 있다. 서열 동일성 백분율은 예를 들면, 비교 영역 전체에서 두 개의 최적으로 정렬된 서열을 비교하는 단계, 두 서열 모두에서 동일한 아미노산 또는 핵산이 나타나는 위치의 개수를 결정하여 일치된(matched) 위치의 개수를 수득하는 단계, 상기 일치된 위치의 개수를 비교 범위 내의 위치의 총 개수(즉, 범위 크기)로 나누는 단계, 및 상기 결과에 100을 곱하여 서열 동일성의 백분율을 수득하는 단계에 의해 계산될 수 있다. 상기 서열 동일성의 퍼센트는 공지의 서열 비교 프로그램을 사용하여 결정될 수 있으며, 일례로 BLASTN(NCBI), CLC Main Workbench (CLC bio), MegAlignTM(DNASTAR Inc) 등을 들 수 있다. As used herein, the term "sequence identity" refers to the degree of identity of amino acid residues or bases between sequences after aligning both sequences in a specific comparison region so as to match each other as much as possible. Sequence identity is a value measured by optimally aligning and comparing two sequences in a specific comparison region, and a part of the sequence in the comparison region may be added or deleted compared to a reference sequence. Percentage sequence identity can be calculated by, for example, comparing two optimally aligned sequences across the comparison region, determining the number of positions in which identical amino acids or nucleic acids occur in both sequences to obtain the number of matched positions. dividing the number of matched positions by the total number of positions within the comparison range (i.e., range size), and multiplying the result by 100 to obtain a percentage of sequence identity. The percent of sequence identity can be determined using a known sequence comparison program, examples of which include BLASTN (NCBI), CLC Main Workbench (CLC bio), MegAlign™ (DNASTAR Inc), and the like.
상기 버크홀데리아 비에트나미엔시스(Burkholderia vietnamiensis) 균주는 기탁번호 KCCM12821P로 기탁된 균주일 수 있다. The Burkholderia vietnamiensis strain may be a strain deposited under accession number KCCM12821P.
상기 균주는 모발 또는 두피 상태 개선 효과를 갖는 것일 수 있다. 예를 들어, 상기 모발 또는 두피 상태 개선은 발모 촉진, 탈모 방지, 모근 강화, 두피 지질 억제, 비듬 방지, 및 두피 염증 억제 중 1종 이상일 수 있다.The strain may have an effect of improving hair or scalp conditions. For example, the hair or scalp condition improvement may be one or more of hair growth promotion, hair loss prevention, hair root strengthening, scalp lipid inhibition, dandruff prevention, and scalp inflammation inhibition.
상기 균주는 분리된 것일 수 있다. 본 명세서에서 용어 "분리된(isolated)"이란 자연계에 존재하는 것이 아닌 인위적으로 분리하여 이용가능한 것을 의미한다. 상기 균주는 인위적으로 분리되어 상기 기탁번호로 기탁되었다. 상기 균주는 인간의 피부로부터 분리된 것일 수 있다. 상기 균주는 인간의 두피로부터 분리된 것일 수 있다.The strain may be isolated. In this specification, the term “isolated” means something that is artificially separated and available that does not exist in nature. The strain was artificially isolated and deposited under the accession number. The strain may be isolated from human skin. The strain may be isolated from human scalp.
다른 양상은 상기 균주의 파쇄액, 배양액, 또는 배양액의 추출물을 제공한다.Another aspect provides a lysate of the strain, a culture medium, or an extract of the culture medium.
상기 균주의 구체적인 내용은 전술한 바와 같다. Details of the strain are as described above.
본 명세서에서 용어 "파쇄액"은 균주 자체에 대해 화학적 또는 물리적 힘을 가하여 균주의 세포벽을 파쇄하여 얻은 산물을 의미할 수 있다.As used herein, the term "lysate" may refer to a product obtained by crushing the cell wall of a strain by applying chemical or physical force to the strain itself.
본 명세서에서 용어 "배양액"은 "배양 상층액", "조건 배양액" 또는 "조정 배지"와 호환적으로 사용될 수 있고, 버크홀데리아 비에트나미엔시스 균주가 시험관 내에서 성장 및 생존할 수 있도록 영양분을 공급할 수 있는 배지에 상기 균주를 일정기간 배양하여 얻는 상기 균주, 이의 대사물, 여분의 영양분 등을 포함하는 전체 배지를 의미할 수 있다. 또한, 상기 배양액은 균주를 배양하여 얻은 균체 배양액에서 균체를 제거한 배양액을 의미할 수 있다. 한편, 상기 배양액 중 균체를 제거한 액체를 "상등액"이라고도 하며, 배양액을 일정시간 가만히 두어 하층에 가라앉은 부분을 제외한 상층의 액체만을 취하거나, 여과를 통해 균체를 제거하거나, 배양액을 원심분리하여 하부의 침전을 제거하고 상부의 액체만을 취하여 획득할 수 있다. 상기 "균체"는 본 발명의 균주 자체를 의미하는 것으로, 피부 샘플 등으로부터 분리하여 선별한 균주 자체 또는 상기 균주를 배양하여 배양액으로부터 분리한 균주를 포함한다. 상기 균체는 배양액을 원심분리하여 하층에 가라앉은 부분을 취하여 획득할 수 있고, 또는 중력에 의해 배양액의 하층으로 가라앉으므로 일정 시간 동안 가만히 두었다가 상부의 액체를 제거함으로써 획득할 수 있다.As used herein, the term "culture medium" may be used interchangeably with "culture supernatant", "conditioned culture medium" or "conditioned medium", and provides nutrients so that Burkholderia bietnamiensis strains can grow and survive in vitro. It may mean the entire medium including the strain obtained by culturing the strain for a certain period of time in a medium that can be supplied, metabolites thereof, and extra nutrients. In addition, the culture solution may mean a culture solution obtained by removing the cells from the cell culture solution obtained by culturing the strain. On the other hand, the liquid from which the cells are removed from the culture solution is also called "supernatant", and the culture solution is left still for a certain period of time to take only the liquid of the upper layer except for the part sunk in the lower layer, remove the cells through filtration, or centrifuge the culture solution to It can be obtained by removing the precipitate and taking only the upper liquid. The "cell" refers to the strain itself of the present invention, and includes the strain itself isolated and selected from a skin sample or the like, or a strain isolated from a culture medium by culturing the strain. The cells can be obtained by centrifuging the culture solution and taking the part that has sunk to the lower layer, or it can be obtained by leaving it for a certain period of time and then removing the upper liquid because it sinks to the lower layer of the culture medium by gravity.
상기 배양액은 균주를 배양하여 수득된 배양액 자체, 그의 농축물 또는 동결건조물, 또는 배양액로부터 균주를 제거하여 수득된 배양 상층액, 그의 농축물 또는 동결건조물을 포함할 수 있다. The culture solution may include the culture solution itself obtained by culturing the strain, its concentrate or lyophilisate, or the culture supernatant obtained by removing the strain from the culture solution, its concentrate or lyophilisate.
상기 배양액은 버크홀데리아 비에트나미엔시스 균주를 배지(예를 들면, R2A 배지)에서 10℃ 초과 또는 40℃ 미만 중 어느 온도에서 일정 시간, 예를 들면, 4 내지 50시간 동안 배양하여 수득된 것일 수 있다. The culture solution may be obtained by culturing the Burkholderia bietnamiensis strain in a medium (eg, R2A medium) at any temperature above 10 ° C or below 40 ° C for a certain period of time, for example, 4 to 50 hours. there is.
일 구체예에서, 균주의 배양 상층액은 균주 배양액을 원심분리나 여과시켜 균주를 제거하는 단계에 의해 수득될 수 있다.In one embodiment, the culture supernatant of the strain may be obtained by centrifuging or filtering the strain culture medium to remove the strain.
다른 구체예에서, 농축물은 상기 균주 배양액 자체, 또는 상기 배양액을 원심분리나 필터를 이용하여 여과한 후 수득된 상층액을 농축하는 단계에 의해 수득될 수 있다. In another embodiment, the concentrate may be obtained by concentrating the supernatant obtained after filtering the strain culture medium itself, or the culture medium using a centrifugal separation or filter.
상기 버크홀데리아 비에트나미엔시스 균주를 배양하기 위한 배양용 배지 및 배양 조건은 통상의 지식을 가진 자가 적절하게 선택하거나 변형하여 이용할 수 있다.The culture medium and culture conditions for culturing the Burkholderia bietnamiensis strain can be appropriately selected or modified by those skilled in the art.
본 명세서에서 용어 "배양액 추출물"은 상기 배양액 또는 그의 농축액으로부터 추출한 것을 의미하며, 추출액, 추출액의 희석액 또는 농축액, 추출액을 건조하여 얻어지는 건조물, 또는 이들 조정제물 또는 정제물, 이를 분획한 분획물을 포함할 수 있다. As used herein, the term "culture broth extract" refers to an extract obtained from the culture medium or a concentrate thereof, and may include an extract, a dilution or concentrate of the extract, a dried product obtained by drying the extract, or a crude or purified product thereof, or a fraction obtained by fractionating the same. can
또 다른 양상은 버크홀데리아 비에트나미엔시스 균주, 상기 균주의 파쇄액, 배양액, 또는 배양액의 추출물의 용도를 제공한다. 상세하게는, 버크홀데리아 비에트나미엔시스 균주, 그의 파쇄액, 배양액 또는 배양액의 추출물을 포함하는 조성물을 제공한다. Another aspect provides the use of a Burkholderia bietnamiensis strain, a lysate, culture, or extract of the strain. Specifically, a composition containing a Burkholderia bietnamiensis strain, a lysate thereof, a culture medium, or an extract of the culture medium is provided.
상기 용도 또는 조성물에 있어서, 상기 버크홀데리아 비에트나미엔시스 균주는 상기 일 양상에 따른 버크홀데리아 비에트나미엔시스 균주일 수 있다. 일 구체예에서, 상기 균주는 서열번호 1과 약 95% 이상, 약 96% 이상, 약 97% 이상, 약 98% 이상, 약 99% 이상, 약 99.5% 이상, 또는 약 99.9% 이상의 서열 동일성을 갖는 16S rRNA를 포함하는 것일 수 있다. 다른 구체예에서, 상기 균주는 서열번호 1의 16S rRNA를 포함하는 것일 수 있다. 또 다른 구체예에서, 상기 균주는 기탁번호 KCCM12821P로 기탁된 균주일 수 있다.In the use or composition, the Burkholderia bietnamiensis strain may be the Burkholderia bietnamiensis strain according to the above aspect. In one embodiment, the strain has at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9% sequence identity with SEQ ID NO: 1 It may include 16S rRNA having In another embodiment, the strain may be one containing the 16S rRNA of SEQ ID NO: 1. In another embodiment, the strain may be a strain deposited with accession number KCCM12821P.
상기 균주의 용도는 모발 또는 두피 상태 개선을 포함할 수 있다.Uses of the strain may include improving hair or scalp condition.
상기 모발 또는 두피 상태 개선은 발모 촉진, 탈모 방지, 모근 강화, 두피 지질 억제, 비듬 방지, 및 두피 염증 억제 중 1종 이상일 수 있다.The hair or scalp condition improvement may be one or more of hair growth promotion, hair loss prevention, hair root strengthening, scalp lipid inhibition, dandruff prevention, and scalp inflammation inhibition.
본 명세서에서 용어 "발모"란 모발이 생성 및 성장하는 것을 의미한다.As used herein, the term "hair growth" means that hair is generated and grown.
본 명세서에서 용어 "발모 촉진"은 "육모 촉진"과 혼용되며, 모발의 생성 및 성장을 촉진하여 전체 모발에서 성장기 모발의 비중을 증가시키는 것을 의미한다. 이는 새로운 모발의 생성을 촉진하는 것뿐만 아니라, 기존 모발이 건강하게 자라도록 하는 것도 포함한다.In this specification, the term "hair growth promotion" is used interchangeably with "hair growth promotion", and means to increase the specific gravity of anagen hair in total hair by promoting hair generation and growth. This includes not only stimulating new hair growth, but also allowing existing hair to grow healthily.
본 명세서에서 용어 "탈모(alopecia)"란 두피로부터 모발이 탈락하는 현상 또는 모발이 성기거나 가늘어지는 상태를 의미한다. 상기 탈모는 성장기 모발의 비중이 감소함에 따라 유발될 수 있다. 상기 탈모는 외인성 요인 또는 내인성 요인에 의해 유발될 수 있다. 상기 외인성 요인은 여러 가지 외부 요인, 예컨대 자외선을 말하고, 상기 내인성 요인은 시간의 흐름에 의해 발생하는 요인으로 텔로미어에 의한 모유두세포의 세포사와 세포 기능 저하에 따른 산화 스트레스 등을 포함한다. 상기 탈모는 원형 탈모증(alopecia areata), 유전성 안드로겐 탈모증(androgenetic alopecia), 휴지기 탈모증(Telogen effluvium), 외상성 탈모증, 발모벽(trichotillomania)으로 인한 탈모, 압박성 탈모증(pressure alopecia), 생장기 탈모증, 비강성 탈모증, 매독성 탈모증(alopecia syphlltiac), 지루 탈모증(alopecia seborrhecia), 증후성 탈모증, 반흔성 탈모증, 및 선천성 탈모증 중에서 선택된 하나 이상일 수 있으나, 이에 제한되지 않는다.As used herein, the term "alopecia" refers to a phenomenon in which hair falls off from the scalp or a condition in which hair becomes sparse or thin. The hair loss may be caused by a decrease in the specific gravity of anagen hair. The hair loss may be caused by extrinsic factors or endogenous factors. The extrinsic factors refer to various external factors, such as ultraviolet rays, and the endogenous factors include oxidative stress due to cell death of dermal papilla cells caused by telomeres and cell function degradation, etc., which occur over time. The hair loss is alopecia areata, hereditary androgenetic alopecia, telogen effluvium, traumatic alopecia, hair loss due to trichotillomania, pressure alopecia, anagen alopecia, pityriasis It may be one or more selected from alopecia, syphilitic alopecia (alopecia syphlltiac), seborrheic alopecia (alopecia seborrhecia), symptomatic alopecia, scarring alopecia, and congenital alopecia, but is not limited thereto.
본 명세서에서 용어 "탈모 방지"란 탈모를 예방하거나 억제하는 것을 의미한다. 따라서, 상기 탈모 방지는 탈모 개선, 탈모 예방, 탈모 치료 등을 포함하는 의미일 수 있다.As used herein, the term "anti-hair loss" means to prevent or suppress hair loss. Therefore, the prevention of hair loss may include hair loss improvement, hair loss prevention, hair loss treatment, and the like.
본 명세서에서 용어 "모근 강화"란 모발의 모근부를 강하고 튼튼하게 하는 모든 작용을 의미한다. 예를 들어, 모유두세포의 증식, 모유두세포로의 영양분 전달 촉진, 모유두세포에서 성장인자의 발현 증가에 의해 모근이 강화될 수 있다.In this specification, the term "hair root strengthening" refers to all actions that strengthen and strengthen the hair root part of the hair. For example, hair roots can be strengthened by proliferation of dermal papilla cells, promotion of nutrient delivery to dermal papilla cells, and increased expression of growth factors in dermal papilla cells.
본 명세서에서 용어 "두피 지질"이란 두피에서 생성된 지질을 의미한다. 지질이 두피에 과다하게 쌓이면 번들거림, 비듬, 따가움, 가려움 등의 문제를 일으킨다. 피지가 과다 분비되면 두피에 지질막을 형성하게 된다. 두피에 지질막이 과다하게 형성되면 혐기성 세균의 번식, 지방산 생성이 증가하고, 이는 각종 트러블, 지루성 두피염, 악취, 탈모 등을 일으킬 수 있다. 상기 지질은 과산화지질을 의미할 수 있다.As used herein, the term "scalp lipid" refers to lipids produced in the scalp. Excessive accumulation of lipids on the scalp causes problems such as shine, dandruff, stinging, and itching. When sebum is excessively secreted, a lipid film is formed on the scalp. When the lipid film is excessively formed on the scalp, the propagation of anaerobic bacteria and the production of fatty acids increase, which can cause various troubles, seborrheic scalpitis, odor, hair loss, and the like. The lipid may mean a lipid peroxide.
본 명세서에서 용어 "두피 지질 억제"란 두피에서 지질의 생성을 감소 또는 저해하는 모든 작용을 의미한다. 두피 지질 억제를 통해, 비듬 방지, 피지로 인한 두피 염증 및 가려움증 개선 또는 억제, 과산화지질로 인한 탈모의 방지 효과를 얻을 수 있다.As used herein, the term "suppressing scalp lipids" refers to any action that reduces or inhibits the production of lipids in the scalp. Prevention of dandruff, improvement or suppression of scalp inflammation and itching due to sebum, and prevention of hair loss due to lipid peroxide can be obtained through inhibition of scalp lipids.
본 명세서에서 용어 "비듬(dandruff)"이란 두피에서 표피 탈락이 발생하여 각질이 눈에 띄게 나타나는 현상을 말한다. 비듬은 가려움증을 동반할 수 있다.In this specification, the term "dandruff" refers to a phenomenon in which epidermal detachment occurs on the scalp and dead skin cells appear conspicuously. Dandruff can be accompanied by itching.
본 명세서에서 용어 "비듬 방지"란 비듬의 생성을 예방하거나 억제하는 것을 의미한다. 따라서, 상기 비듬 방지는 비듬 개선, 비듬 예방 등을 포함하는 의미일 수 있다.As used herein, the term "anti-dandruff" means preventing or inhibiting the production of dandruff. Therefore, the anti-dandruff may mean improving dandruff, preventing dandruff, and the like.
본 명세서에서 용어 "두피 염증"이란 두피에 발생한 염증을 의미한다. 상기 두피 염증은 지루성 피부염, 소양증, 건성 습진, 홍반, 두드러기, 건선, 약발진, 두피 여드름 중 어느 하나 이상을 포함할 수 있다.As used herein, the term "scalp inflammation" means inflammation occurring in the scalp. The scalp inflammation may include any one or more of seborrheic dermatitis, pruritus, dry eczema, erythema, urticaria, psoriasis, drug rash, and scalp acne.
본 명세서에서 용어 "두피 염증 억제"란 두피 염증을 감소 또는 개선시키거나, 두피 염증의 발생을 예방 또는 억제하는 모든 작용을 의미한다. As used herein, the term “suppressing scalp inflammation” refers to any action that reduces or improves scalp inflammation or prevents or suppresses the occurrence of scalp inflammation.
본 명세서에서 용어 "예방"은 질병의 발생을 억제하는 것을 포함한다.As used herein, the term "prevention" includes inhibiting the occurrence of a disease.
본 명세서에서 용어 "치료"는 질병의 발전의 억제, 경감, 또는 제거를 포함한다.As used herein, the term "treatment" includes the inhibition, alleviation, or elimination of the development of a disease.
일 양상에 따른 조성물은 버크홀데리아 비에트나미엔시스 균주, 그의 파쇄액, 배양액 또는 배양액의 추출물을 포함함으로써 인간모유두세포(Human follicular dermal papilla cells, HFDPC)를 증식시킬 수 있다. 따라서, 상기 조성물은 모근 강화 효과, 탈모 방지 효과, 및 발모 촉진 효과를 나타낼 수 있다.The composition according to one aspect may proliferate human follicular dermal papilla cells (HFDPC) by including a Burkholderia bietnamiensis strain, a lysate thereof, a culture medium, or an extract of the culture medium. Therefore, the composition may exhibit a hair root strengthening effect, an hair loss preventing effect, and a hair growth promoting effect.
일 양상에 따른 조성물은 버크홀데리아 비에트나미엔시스 균주, 그의 파쇄액, 배양액 또는 배양액의 추출물을 포함함으로써 두피 지질을 억제시킬 수 있다. 따라서, 상기 조성물은 두피 지질 억제를 통해, 비듬 방지, 피지로 인한 두피 염증 및 가려움증 개선 또는 억제, 과산화지질로 인한 탈모의 방지 효과를 나타낼 수 있다.The composition according to one aspect may inhibit scalp lipids by including a Burkholderia bietnamiensis strain, a lysate thereof, a culture medium, or an extract of the culture medium. Therefore, the composition may exhibit effects of preventing dandruff, improving or suppressing scalp inflammation and itching due to sebum, and preventing hair loss due to lipid peroxide through inhibition of scalp lipids.
또 다른 구체예에 있어서, 상기 균주는 모발 또는 두피 상태 개선 효과를 갖는 다른 균주, 예를 들어, 버크홀데리아 속에 속하는 균주와 함께 사용되어 시너지 효과를 나타낼 수 있다.In another embodiment, the strain may exhibit a synergistic effect when used together with other strains having an effect of improving hair or scalp conditions, for example, strains belonging to the genus Burkholderia.
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 균주, 이의 파쇄액, 배양액, 또는 배양액의 추출물을 포함할 수 있다.The composition is 0.001% to 80% by weight, for example, 0.01% to 60% by weight, 0.01% to 40% by weight, 0.01% to 30% by weight, 0.01% to 20% by weight, based on the total weight of the composition. %, 0.01% to 10%, 0.01% to 5%, 0.05% to 60%, 0.05% to 40%, 0.05% to 30%, 0.05% to 20%, 0.05% to 10% by weight, 0.05% to 5% by weight, 0.1% to 60% by weight, 0.1% to 40% by weight, 0.1% to 30% by weight, 0.1% to 20% by weight, 0.1% by weight % to 10% by weight, or 0.1% to 5% by weight of the strain, a lysate thereof, a culture medium, or an extract of the culture medium.
본 명세서에서 용어, "유효성분으로 포함"은 상기에서 언급한 효과를 나타낼 수 있는 정도로 본 명세서의 균주, 이의 파쇄액, 배양액, 또는 배양액의 추출물이 첨가되는 것을 의미하고, 약물전달 및 안정화 등을 위하여 다양한 성분을 부성분으로 첨가하여 다양한 형태로 포뮬레이션(formulation)되는 것을 포함하는 의미이다.As used herein, the term "included as an active ingredient" means that the strain of the present specification, its lysate, culture medium, or extract of the culture medium is added to the extent that the above-mentioned effects can be exhibited, and drug delivery and stabilization, etc. It means that it is formulated in various forms by adding various components as subcomponents for the purpose.
상기 조성물은 액체 상태 또는 건조 상태인 것일 수 있다. 일 구체예에서, 상기 조성물은 건조 분말 형태일 수 있다.The composition may be in a liquid or dry state. In one embodiment, the composition may be in the form of a dry powder.
상기 조성물을 건조 상태로 제조하는 건조 방법은 당업계에서 일반적으로 사용되는 방법을 사용할 수 있으며, 특별히 제한되지 않는다. 상기 건조 방법의 비제한적인 예는, 공기 건조 방법, 자연 건조 방법, 분무 건조 방법, 동결 건조 방법 등이 있다. 이들 방법은 단독으로 사용하거나 적어도 두 가지 방법을 함께 사용할 수 있다.A drying method for preparing the composition in a dry state may use a method commonly used in the art, and is not particularly limited. Non-limiting examples of the drying method include an air drying method, a natural drying method, a spray drying method, a freeze drying method, and the like. These methods may be used alone or at least two methods may be used together.
상기 조성물은 상기 균주, 그의 파쇄액, 배양액 또는 배양액의 추출물의 열화(deterioration)를 감소시키기에 충분한 유효량의 첨가제를 포함할 수 있다. 상기 첨가제는 예를 들어 결합제일 수 있으나, 이에 제한되지 않는다.The composition may contain an additive in an effective amount sufficient to reduce deterioration of the strain, its lysate, culture medium or extract of the culture broth. The additive may be, for example, a binder, but is not limited thereto.
상기 조성물은 화장품학적으로, 약학적으로 또는 식품학적으로 허용가능한 담체를 더 포함할 수 있다. 상기 조성물은 담체와 함께 제형화되어 화장품, 약품, 식품 첨가제 등으로 제공될 수 있다. The composition may further include a cosmetically, pharmaceutically or food-acceptable carrier. The composition may be formulated with a carrier and provided as cosmetics, drugs, food additives, and the like.
상기 조성물은 화장료 조성물일 수 있다.The composition may be a cosmetic composition.
상기 화장료 조성물은 본 명세서에 개시된 유효성분 이외에 화장료 조성물에 통상적으로 이용되는 성분들, 기능성 첨가물 등을 추가로 포함할 수 있으며, 예컨대 항산화제, 안정화제, 용해화제, 계면활성제, 분산제, 방부제, 비타민, 안료, 향료 등과 같은 통상적인 보조제, 그리고 담체를 포함할 수 있다. The cosmetic composition may further include ingredients commonly used in cosmetic compositions, functional additives, etc. in addition to the active ingredients disclosed herein, such as antioxidants, stabilizers, solubilizers, surfactants, dispersants, preservatives, and vitamins. , conventional adjuvants such as pigments, flavors, and the like, and carriers.
상기 화장료 조성물은 특정 제형에 특별히 한정되지 않으며, 목적하는 바에 따라 제형을 적절히 선택할 수 있다. 상기 화장료 조성물은, 예를 들어, 가용화 제형, 유화 제형, 또는 분산 제형을 갖는 것일 수 있다. 상기 화장료 조성물은, 예를 들어, 유연화장수, 영양화장수, 마사지크림, 영양크림, 에센스, 팩, 젤, 앰플 또는 피부 점착 타입의 화장료 제형을 갖는 것일 수 있다.The cosmetic composition is not particularly limited to a specific formulation, and the formulation may be appropriately selected according to the purpose. The cosmetic composition may have, for example, a solubilizing formulation, an emulsifying formulation, or a dispersion formulation. The cosmetic composition may have, for example, a softening lotion, a nutrient lotion, a massage cream, a nutrient cream, an essence, a pack, a gel, an ampoule, or a skin-adhesive cosmetic formulation.
상기 화장료 조성물은 모발 또는 두피에 사용할 수 있는 제형일 수 있다. 상기 화장료 조성물은, 예를 들면 헤어토닉, 헤어컨디셔너, 헤어에센스, 헤어로션, 헤어영양로션, 헤어샴푸, 헤어린스, 헤어트리트먼트, 헤어크림, 헤어영양크림, 헤어모이스처크림, 헤어맛사지크림, 헤어왁스, 헤어 에어로졸, 헤어팩, 헤어영양팩, 헤어비누, 헤어클렌징폼, 머릿기름, 모발건조제, 모발보존처리제, 모발염색제, 모발용 웨이브제, 모발탈색제, 헤어겔, 헤어글레이즈, 헤어드레싱어, 헤어래커, 헤어모이스처라이저, 헤어무스 또는 헤어스프레이의 제형일 수 있고, 구체적으로 샴푸, 클렌져, 컨디셔너, 트리트먼트, 헤어팩, 린스, 두피팩, 토닉, 에센스, 로션, 크림, 오일, 미스트, 스프레이, 또는 헤어젤 제형일 수 있다.The cosmetic composition may be a formulation that can be used for hair or scalp. The cosmetic composition, for example, hair tonic, hair conditioner, hair essence, hair lotion, hair nutrition lotion, hair shampoo, hair rinse, hair treatment, hair cream, hair nutrition cream, hair moisture cream, hair massage cream, hair Wax, hair aerosol, hair pack, hair nutrition pack, hair soap, hair cleansing foam, hair oil, hair drying agent, hair preservative, hair dye, hair waving agent, hair bleach, hair gel, hair glaze, hair dressing, hair lacquer, It may be in the form of a hair moisturizer, hair mousse or hair spray, specifically shampoo, cleanser, conditioner, treatment, hair pack, rinse, scalp pack, tonic, essence, lotion, cream, oil, mist, spray, or hair gel may be a formulation.
상기 조성물은 피부외용제용 조성물일 수 있다. The composition may be a composition for external application for skin.
본 명세서에서, 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 염 등의 약제; 비타민 C, 마그네슘아스코빌포스페이트, 아스코르브산글루코시드, 알부틴, 코지산; 글루코스, 프룩토스, 트레할로스 등의 당류 등도 적절하게 배합할 수 있다.In the present specification, the external skin preparation may be a cream, gel, ointment, skin emulsifier, skin suspension, transdermal delivery patch, drug-containing bandage, lotion, or a combination thereof. The external skin preparation is a component usually used in external preparations for skin such as cosmetics or pharmaceuticals, for example, water-based components, oil-based components, powder components, alcohols, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, and fragrances. , colorants, various skin nutrients, or combinations thereof and may be suitably blended as needed. The external skin preparation may include metal sequestering agents such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, and gluconic acid; drugs such as caffeine, tannin, bellapamil, licorice extract, glabridin, hot water extract of calin fruit, various herbal medicines, tocopherol acetate, glycylitnic acid, tranexamic acid and its derivatives or salts; Vitamin C, magnesium ascorbyl phosphate, ascorbic acid glucoside, arbutin, kojic acid; Sugars, such as glucose, fructose, and trehalose, etc. can also be mix|blended suitably.
상기 조성물은 약학적 조성물일 수 있다. The composition may be a pharmaceutical composition.
상기 약학적 조성물은 약제학적으로 허용가능한 희석제 또는 담체를 추가적으로 포함할 수 있다. 상기 희석제는 유당, 옥수수 전분, 대두유, 미정질 셀룰로오스, 만니톨, 또는 그 조합일 수 있다. 상기 담체는 부형제, 붕해제, 결합제, 활택제, 또는 그 조합일 수 있다. 상기 부형제는 미결정 셀룰로오즈, 유당, 저치환도 히드록시셀룰로오즈, 또는 그 조합일 수 있다. 상기 붕해제는 카르복시메틸셀룰로오스 칼슘, 전분글리콜산 나트륨, 무수인산일수소 칼슘, 또는 그 조합일 수 있다. 상기 결합제는 폴리비닐피롤리돈, 저치환도 히드록시프로필셀룰로오즈, 히드록시프로필셀룰로오즈, 또는 그 조합일 수 있다. 상기 활택제는 스테아린산 마그네슘, 이산화규소, 탈크, 또는 그 조합일 수 있다.The pharmaceutical composition may additionally include a pharmaceutically acceptable diluent or carrier. The diluent may be lactose, corn starch, soybean oil, microcrystalline cellulose, mannitol, or a combination thereof. The carrier may be an excipient, a disintegrant, a binder, a lubricant, or a combination thereof. The excipient may be microcrystalline cellulose, lactose, low-substituted hydroxycellulose, or a combination thereof. The disintegrant may be calcium carboxymethylcellulose, sodium starch glycolate, calcium monohydrogen phosphate, or a combination thereof. The binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or a combination thereof. The lubricant may be magnesium stearate, silicon dioxide, talc, or a combination thereof.
상기 약학적 조성물은 경구 또는 비경구 투여 제형으로 제형화될 수 있다. 경구 투여 제형은 과립제, 산제, 액제, 정제, 캅셀제, 건조시럽제 등일 수 있다. 비경구 투여 제형은 주사제, 연고제 등일 수 있다.The pharmaceutical composition may be formulated for oral or parenteral administration. Oral dosage forms may be granules, powders, liquids, tablets, capsules, dry syrups, and the like. Parenteral dosage forms may be injections, ointments, and the like.
상기 조성물은 건강기능식품 조성물일 수 있다. The composition may be a health functional food composition.
상기 건강기능식품 조성물은 상기 균주, 그의 파쇄액, 배양액, 또는 배양액의 추출물 단독을 사용하거나, 또는 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효 성분의 혼합 양은 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다. 일반적으로, 식품 또는 음료의 제조시에 본 명세서의 조성물은 원료에 대하여 50 중량부 이하, 또는 15 중량부 이하의 양으로 첨가될 수 있다. 상기 건강기능식품의 종류에는 특별한 제한은 없다. 건강기능식품의 종류 중 음료 조성물은 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상기 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드; 말토스, 슈크로스와 같은 디사카라이드; 및 덱스트린, 사이클로덱스트린과 같은 폴리사카라이드; 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 감미제로서는 타우마틴, 스테비아 추출물과 같은 천연 감미제나, 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다. 상기 건강식품 조성물은 또한 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, 또는 그 조합을 함유할 수 있다. 상기 건강기능식품 조성물은 또한, 천연 과일쥬스, 과일쥬스 음료, 야채 음료의 제조를 위한 과육, 또는 그 조합을 함유할 수 있다.The health functional food composition may be used alone, or in combination with other foods or food ingredients, and may be appropriately used according to conventional methods. The mixing amount of the active ingredient can be suitably determined depending on the purpose of use (prevention, health or therapeutic treatment). In general, when preparing food or beverage, the composition of the present specification may be added in an amount of 50 parts by weight or less, or 15 parts by weight or less based on the raw material. There is no particular limitation on the type of health functional food. Among the types of health functional foods, beverage compositions may contain various flavoring agents or natural carbohydrates as additional components, like conventional beverages. The natural carbohydrates include monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; and polysaccharides such as dextrins and cyclodextrins; It is a sugar alcohol, such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as thaumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame may be used. The health food composition may also contain nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, and carbonated beverages. carbonation agent used, or a combination thereof. The health functional food composition may also contain natural fruit juice, fruit juice beverages, fruit flesh for preparing vegetable beverages, or a combination thereof.
또 다른 양상은 유효한 양의 상기한 조성물을 그를 필요로 하는 개체에 처리 또는 투여하는 단계를 포함하는 개체의 상태를 예방, 개선, 또는 치료하는 방법을 제공한다. Another aspect provides a method for preventing, ameliorating, or treating a condition in a subject comprising treating or administering to a subject in need thereof an effective amount of a composition described above.
상기 개체의 상태는 모발 또는 두피 상태일 수 있고, 구체적으로 탈모와 관련된 상태, 또는 두피 염증과 관련된 상태일 수 있다.The condition of the subject may be a hair or scalp condition, and may specifically be a condition related to hair loss or a condition related to inflammation of the scalp.
본 명세서에서 용어, "투여하는", "도입하는", 및 "이식하는"은 상호교환적으로 사용되고, 일 구체예에 따른 조성물의 원하는 부위로의 적어도 부분적 국소화를 초래하는 방법 또는 경로에 의한 개체 내로의 일 구체예에 따른 조성물의 배치를 의미할 수 있다. The terms "administering," "introducing," and "implanting" are used interchangeably herein, and according to one embodiment, a subject by a method or route that results in at least partial localization of a composition to a desired site. It can mean the placement of a composition according to one embodiment into.
투여는 당업계에 알려진 방법에 의하여 투여될 수 있다. 투여는 예를 들면, 정맥내, 근육내, 경구, 경피(transdermal), 점막, 코안(intranasal), 기관내(intratracheal) 또는 피하 투여와 같은 경로로, 임의의 수단에 의하여 개체로 직접적으로 투여될 수 있다. 상기 투여는 전신적으로 또는 국부적으로 투여될 수 있다.Administration may be administered by a method known in the art. Administration may be administered directly to a subject by any means, for example, by routes such as intravenous, intramuscular, oral, transdermal, mucosal, intranasal, intratracheal or subcutaneous administration. can The administration may be administered systemically or locally.
상기 개체는 포유동물, 예를 들면, 사람, 소, 말, 돼지, 개, 양, 염소, 또는 고양이일 수 있다. 상기 개체는 모발 또는 두피 상태 개선을 필요로 하는 개체, 예를 들어 발모 촉진, 탈모 방지, 모근 강화, 두피 지질 억제, 비듬 방지, 또는 두피 염증 억제 효과를 필요로 하는 개체일 수 있다.The subject may be a mammal, such as a human, cow, horse, pig, dog, sheep, goat, or cat. The subject may be a subject in need of hair or scalp condition improvement, for example, a subject in need of hair growth promotion, hair loss prevention, hair root strengthening, scalp lipid inhibition, dandruff prevention, or scalp inflammation inhibitory effect.
상기 투여는 일 구체예에 따른 조성물을 개체당 일당 0.1 ㎎ 내지 1,000 ㎎, 예를 들면, 0.1 ㎎ 내지 500 ㎎, 0.1 ㎎ 내지 100 ㎎, 0.1 ㎎ 내지 50 ㎎, 0.1 ㎎ 내지 25 ㎎, 1 ㎎ 내지 1,000 ㎎, 1 ㎎ 내지 500 ㎎, 1 ㎎ 내지 100 ㎎, 1 ㎎ 내지 50 ㎎, 1 ㎎ 내지 25 ㎎, 5 ㎎ 내지 1,000 ㎎, 5 ㎎ 내지 500 ㎎, 5 ㎎ 내지 100 ㎎, 5 ㎎ 내지 50 ㎎, 5 ㎎ 내지 25 ㎎, 10 ㎎ 내지 1,000 ㎎, 10 ㎎ 내지 500 ㎎, 10 ㎎ 내지 100 ㎎, 10 ㎎ 내지 50 ㎎, 또는 10 ㎎ 내지 25 ㎎을 투여하는 것일 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 1일 1회 또는 임상적으로 용인가능한 부작용의 범위 내에서 2회 이상이 가능하고, 투여 부위에 대해서도 1개소 또는 2개소 이상에 투여할 수 있으며, 매일 또는 2 내지 5일 간격으로 총 투여 일수는 한번 치료 시 1일에서 30일까지 투여될 수 있다. 필요한 경우, 적정 시기 이후에 동일한 치료를 반복할 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.The administration is 0.1 mg to 1,000 mg per day of the composition according to one embodiment, for example, 0.1 mg to 500 mg, 0.1 mg to 100 mg, 0.1 mg to 50 mg, 0.1 mg to 25 mg, 1 mg to 1,000 mg, 1 mg to 500 mg, 1 mg to 100 mg, 1 mg to 50 mg, 1 mg to 25 mg, 5 mg to 1,000 mg, 5 mg to 500 mg, 5 mg to 100 mg, 5 mg to 50 mg , 5 mg to 25 mg, 10 mg to 1,000 mg, 10 mg to 500 mg, 10 mg to 100 mg, 10 mg to 50 mg, or 10 mg to 25 mg may be administered. However, the dosage may be prescribed in various ways depending on factors such as formulation method, administration method, patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate and reaction sensitivity, and those skilled in the art can Dosage can be appropriately adjusted in consideration of these factors. The number of administrations can be once a day or two or more times within the range of clinically acceptable side effects, and administration can be performed at one or two or more sites, daily or at intervals of 2 to 5 days. The number of administration days may be administered from 1 day to 30 days per treatment. If necessary, the same treatment can be repeated after a titration period. For non-human animals, the same dosage per kg as for humans is used, or the above dosage is converted by the volume ratio (eg, average value) of the organ (heart, etc.) between the target animal and the human. A single dose can be administered.
일 양상에 따른 신규 버크홀데리아 비에트나미엔시스 균주에 의하면, 발모 촉진, 탈모 방지, 모근 강화, 두피 지질 억제, 비듬 방지, 및 두피 염증 억제 효과가 있으므로, 모발 또는 두피 상태의 개선, 예방, 또는 치료 용도로 다양하게 응용될 수 있다.According to the novel Burkholderia bietnamiensis strain according to one aspect, there are effects of promoting hair growth, preventing hair loss, strengthening hair roots, inhibiting scalp lipids, preventing dandruff, and inhibiting scalp inflammation, thereby improving, preventing, or treating hair or scalp conditions. It can be applied for various purposes.
도 1은 모발의 구조 및 모유두세포를 도시한 도이다.
도 2는 모유두세포의 증식능 증가 효능을 확인한 결과이다.
도 3은 각질형성세포에서 분비되는 지질의 발생 감소 효과를 나타낸 것이다.1 is a diagram showing the structure of hair and dermal papilla cells.
2 is a result confirming the efficacy of increasing the proliferation capacity of dermal papilla cells.
Figure 3 shows the effect of reducing the generation of lipids secreted from keratinocytes.
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 through examples. However, these examples are intended to illustrate the present invention by way of example, and the scope of the present invention is not limited to these examples.
실시예 1. 균주의 분리 및 동정Example 1. Isolation and identification of strains
건강한 두피를 가진 20대 남성의 두피를 멸균 증류수로 세척하여 샘플을 얻었다. 상기 샘플을 R2A(Reasoner's 2A) 액체 배지 또는 고체 배지(제조사: Difco)에 접종하였다. 접종 후 48시간 동안 28℃ 인큐베이터에서 배양한 뒤, 형성된 집락 100개를 순수분리 배양하여, 48시간 동안 28℃ 인큐베이터에서 재배양 하였다. 배양이 완료된 집락은 16S rRNA 서열 동정을 실시하였다. 이때 사용된 프라이머는 박테리아에만 반응하여 증폭하도록 고안되었다. PCR 증폭은 95℃ 1분, 55℃ 1분, 75℃ 1분 30초씩 30 사이클로 실시 하였으며, 마지막으로 72℃에서 8분간 유지한 후 4℃에서 보관하였다. PCR 반응이 끝난 뒤 분리 배양된 종들의 DNA 서열은 ABI-3730XL(ABI, USA)를 이용하여 결정하였다. A sample was obtained by washing the scalp of a male in his twenties with a healthy scalp with sterile distilled water. The samples were inoculated into R2A (Reasoner's 2A) liquid medium or solid medium (manufacturer: Difco). After culturing in a 28° C. incubator for 48 hours after inoculation, 100 formed colonies were cultured in pure isolation and re-cultured in a 28° C. incubator for 48 hours. 16S rRNA sequence identification was performed on colonies completed with culture. The primers used at this time were designed to amplify in response only to bacteria. PCR amplification was carried out in 30 cycles of 95 ° C for 1 minute, 55 ° C for 1 minute, and 75 ° C for 1 minute and 30 seconds, and finally maintained at 72 ° C for 8 minutes and then stored at 4 ° C. After the PCR reaction, the DNA sequences of the isolated and cultured species were determined using ABI-3730XL (ABI, USA).
분리 배양된 미생물 집락 중 결정된 16S rRNA 부위의 염기서열을 미국 국립생물정보센터(NCBI, National Center for Biotechnology Information) 홈페이지에서 제공되는 BLAST 프로그램을 통해 등록된 다른 균주들과 비교 분석 하였을 때, 상동성 97% 이하의 신규성 있는 종들만 선별하였고, 그 중 상동성 98% 이하의 신규 미생물 버크홀데리아 비에트나미엔시스(Burkholderia vietnamiensis) 균주(이하 "SP-7"이라 함)를 선별하였다. 선별된 SP-7 균주를 2020년 10월 28일자로 한국 미생물 보존센터에 기탁하여 기탁번호 KCCM12821P를 부여 받았다. SP-7 균주는 서열번호 1(complementary DNA)의 16S rRNA 서열을 갖는다.When the nucleotide sequence of the 16S rRNA region determined in the isolated and cultured microbial colony was compared and analyzed with other strains registered through the BLAST program provided on the homepage of the National Center for Biotechnology Information (NCBI), homology 97 Only novel species of less than % were selected, and among them, a novel microorganism Burkholderia vietnamiensis strain (hereinafter referred to as "SP-7") having 98% or less homology was selected. The selected SP-7 strain was deposited with the Korea Microorganism Conservation Center on October 28, 2020 and was given accession number KCCM12821P. The SP-7 strain has a 16S rRNA sequence of SEQ ID NO: 1 (complementary DNA).
실험예 1. 인간모유두세포(HFDPC) 증식능 증가 효능 평가Experimental Example 1. Evaluating the efficacy of increasing the proliferative capacity of human dermal papilla cells (HFDPC)
실시예 1에서 분리된 SP-7 균주의 배양액이 모유두세포의 증식을 현저하게 유도하는지를 평가하기 위해, 다음과 조건으로 MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) 분석을 수행하였다. In order to evaluate whether the culture of the SP-7 strain isolated in Example 1 markedly induces the proliferation of dermal papilla cells, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyl-2H-tetrazolium bromide) assay was performed.
먼저 인간모유두세포(HFDPC)를 96웰 플레이트에 2x104 세포/웰씩 분주한 후, 37℃, 5% CO2 조건의 배양기에서 24시간 동안 배양하였다. 이후, 아무것도 포함하지 않은 배지로 교체한 후, 상기 SP-7 균주의 배양액 1%(w/w)를 첨가하고 24시간 동안 추가 배양하였다. 균주 배양액 대신 모발 발육 촉진제로 알려진 미녹시딜(Minoxidil, MXD, 6-piperidin-1-ylpyrimidine-2,4-diamine 3-oxide)을 10ppm 첨가한 군을 양성 대조군으로 설정하고, 균주 배양액을 포함하지 않는 배지 첨가군(배지con)을 음성 대조군으로 설정하였으며, 균주 배양액의 효능을 확인하기 위해 배양액 비첨가군(None)도 대조군으로 설정하였다. 24시간 후, 각 웰의 배지를 제거하고 DPBS (Dulbecco's phosphate buffered saline)를 이용한 워싱 과정을 1회 거친 다음, 각 웰당 500μl의 0.5mg/mL MTT (Sigma-aldrich, St. Louis, USA)를 포함하는 배지로 교체하고 37℃에서 4시간 동안 반응시켰다. 이후 빅터 3 (Victor 3, Perkin-Elmer)를 이용하여 570nm에서 흡광도를 측정하였다. 측정된 평균 흡광도 값에 대한 백분율로 세포의 생존율을 나타내었으며, 이를 통해 인간모유두세포(HFDPC) 증식능을 확인할 수 있다. First, human dermal papilla cells (HFDPC) were dispensed in 2x10 4 cells/well in a 96-well plate, and then cultured for 24 hours in an incubator at 37°C and 5% CO 2 conditions. Then, after replacing with a medium containing nothing, 1% (w / w) of the culture medium of the SP-7 strain was added and further cultured for 24 hours. A group containing 10 ppm of minoxidil (Minoxidil, MXD, 6-piperidin-1-ylpyrimidine-2,4-diamine 3-oxide), known as a hair growth promoter, was set as a positive control group instead of the strain culture medium, and a medium containing no strain culture medium The addition group (medium con) was set as a negative control group, and the culture medium addition group (None) was also set as a control group to confirm the efficacy of the strain culture medium. After 24 hours, the medium of each well was removed, washed once using DPBS (Dulbecco's phosphate buffered saline), and then 500μl of 0.5mg/mL MTT (Sigma-aldrich, St. Louis, USA) was added to each well. medium and reacted at 37°C for 4 hours. Then, absorbance was measured at 570 nm using Victor 3 (Perkin-Elmer). The cell viability was expressed as a percentage of the measured average absorbance value, and through this, the proliferative ability of human dermal papilla cells (HFDPC) can be confirmed.
도 2는 모유두세포의 증식능 증가 효능을 확인한 결과이다.2 is a result confirming the efficacy of increasing the proliferation capacity of dermal papilla cells.
도 2에 나타낸 바와 같이, 실시예 1의 균주는 우수한 모유두세포 증식 효과를 나타내는 것을 확인할 수 있었다.As shown in Figure 2, it was confirmed that the strain of Example 1 exhibits an excellent dermal papilla cell proliferation effect.
따라서, SP-7 균주는 모근 강화, 탈모 방지, 및 발모 촉진을 위해 사용될 수 있음을 확인하였다.Therefore, it was confirmed that the SP-7 strain can be used for strengthening hair roots, preventing hair loss, and promoting hair growth.
실험예 2. 두피 지질 억제 효능 평가Experimental Example 2. Scalp lipid inhibition efficacy evaluation
실시예 1에서 분리된 SP-7 균주의 배양액이 두피 지질을 억제하는지 여부를 평가하기 위해, 각질형성세포에 과산화지질을 유도한 후 지질 억제 평가를 수행하였다. In order to evaluate whether the culture solution of the SP-7 strain isolated in Example 1 inhibits scalp lipids, lipid inhibition was evaluated after inducing lipid peroxides into keratinocytes.
구체적으로, 인간각질형성세포(HaCaT)를 6웰 플레이트에 3x105 세포/웰씩 분주한 후, 세포 배양조건에서 24시간 동안 배양하였다. 24시간 이후 배지를 버리고 PBS로 세척한 다음, FBS를 포함하지 않는 배지에 과산화스쿠알렌(peroxidated squalene)을 1%(w/w)로 희석하고, 이를 세포에 첨가하여 과산화지질을 유도하였다. 이후, SP-7 균주의 배양액 1%(w/w)을 첨가하고 72시간 동안 추가 배양하였다. 균주 배양액을 포함하지 않는 배지 1%(w/w) 첨가군(배지con, RCM control)을 음성 대조군으로 설정하였다. 배지를 버리고 PBS로 3회 세척한 다음, 세포의 지질을 염색하기 위해 Nile red O 염색(staining)을 실시하였다. FBS를 포함하지 않는 배지에 Nile red 용액(아세톤 중 1 mg/ml)을 1/1000로 희석하여 세포에 첨가하고, 10분 후 IncucyteZoom(Essen BioScience) 기기에 넣어 형광 현미경으로 촬영하고 형광 세기를 정량하였다. Specifically, after dispensing human keratinocytes (HaCaT) into a 6-well plate by 3x10 5 cells/well, they were cultured for 24 hours under cell culture conditions. After 24 hours, the medium was discarded, washed with PBS, and peroxidated squalene was diluted to 1% (w/w) in a medium without FBS, and lipid peroxide was induced by adding it to the cells. Thereafter, 1% (w/w) of the culture medium of the SP-7 strain was added and further cultured for 72 hours. A medium 1% (w/w) addition group (media con, RCM control) not containing the strain culture medium was set as a negative control group. After discarding the medium and washing with PBS three times, Nile red O staining was performed to stain cell lipids. Nile red solution (1 mg/ml in acetone) diluted 1/1000 in a medium without FBS was added to the cells, and after 10 minutes, it was put into an IncucyteZoom (Essen BioScience) device and photographed with a fluorescence microscope, and fluorescence intensity was quantified. did
도 3은 각질형성세포에서 분비되는 지질의 발생 감소 효과를 나타낸 것이다.Figure 3 shows the effect of reducing the generation of lipids secreted from keratinocytes.
도 3에 나타낸 바와 같이, 실시예 1의 균주는 우수한 지질 억제 효과를 나타내는 것을 확인할 수 있었다.As shown in Figure 3, it was confirmed that the strain of Example 1 exhibits an excellent lipid inhibition effect.
따라서, SP-7 균주는 두피 지질 억제, 비듬 방지, 피지로 인한 두피 염증 및 가려움증 개선 또는 억제, 과산화지질로 인한 탈모의 방지, 및 두피 건강을 위해 사용될 수 있음을 확인하였다.Therefore, it was confirmed that the SP-7 strain can be used for suppressing scalp lipids, preventing dandruff, improving or suppressing scalp inflammation and itching caused by sebum, preventing hair loss due to lipid peroxides, and promoting scalp health.
이상의 설명은 본 발명의 기술 사상을 예시적으로 설명한 것에 불과한 것으로서, 본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자라면 본 발명의 본질적인 특성에서 벗어나지 않는 범위에서 다양한 수정 및 변형이 가능할 것이다. The above description is merely an example of the technical idea of the present invention, and various modifications and variations can be made to those skilled in the art without departing from the essential characteristics of the present invention.
SEQUENCE LISTING <110> Cosmax, INC. <120> Burkholderia vietnamiensis strain and its use for improving hair or scalp condition <130> PN136751 <160> 1 <170> PatentIn version 3.2 <210> 1 <211> 1364 <212> DNA <213> Burkholderia vietnamiensis <400> 1 cacgggtgct tgcacctggt ggcgagtggc gaacgggtga gtaatacatc ggaacatgtc 60 ctgtagtggg ggatagcccg gcgaaagccg gattaatacc gcatacgatc tatggatgaa 120 agcgggggac cttcgggcct cgcgctatag ggttggccga tggctgatta gctagttggt 180 ggggtaaagg cctaccaagg cgacgatcag tagctggtct gagaggacga ccagccacac 240 tgggactgag acacggccca gactcctacg ggaggcagca gtggggaatt ttggacaatg 300 ggcgaaagcc tgatccagca atgccgcgtg tgtgaagaag gccttcgggt tgtaaagcac 360 ttttgtccgg aaagaaatcc ttggctctaa tacagtcggg ggatgacggt accggaagaa 420 taagcaccgg ctaactacgt gccagcagcc gcggtaatac gtagggtgca agcgttaatc 480 ggaattactg ggcgtaaagc gtgcgcaggc ggtttgctaa gaccgatgtg aaatccccgg 540 gctcaacctg ggaactgcat tggtgactgg caggctagag tatggcagag gggggtagaa 600 ttccacgtgt agcagtgaaa tgcgtagaga tgtggaggaa taccgatggc gaaggcagcc 660 ccctgggcca atactgacgc tcatgcacga aagcgtgggg agcaaacagg attagatacc 720 ctggtagtcc acgccctaaa cgatgtcaac tagttgttgg ggattcattt ccttagtaac 780 gtagctaacg cgtgaagttg accgcctggg gagtacggtc gcaagattaa aactcaaagg 840 aattgacggg gacccgcaca agcggtggat gatgtggatt aattcgatgc aacgcgaaaa 900 accttaccta cccttgacat ggtcggaatc ctgaagagat tcgggagtgc tcgaaagaga 960 accggcgcac aggtgctgca tggctgtcgt cagctcgtgt cgtgagatgt tgggttaagt 1020 cccgcaacga gcgcaaccct tgtccttagt tgctacgcaa gagcactcta aggagactgc 1080 cggtgacaaa ccggaggaag gtggggatga cgtcaagtcc tcatggccct tatgggtagg 1140 gcttcacacg tcatacaatg gtcggaacag agggttgcca acccgcgagg gggagctaat 1200 cccagaaaac cgatcgtagt ccggattgca ctctgcaact cgagtgcatg aagctggaat 1260 cgctagtaat cgcggatcag catgccgcgg tgaatacgtt cccgggtctt gtacacaccg 1320 cccgtcacac catgggagtg ggttttacca gaagtggcta gtct 1364 SEQUENCE LISTING <110> Cosmax, INC. <120> Burkholderia vietnamiensis strain and its use for improving hair or scalp condition <130> PN136751 <160> 1 <170> PatentIn version 3.2 <210> 1 <211> 1364 <212> DNA <213> Burkholderia vietnamiensis <400> 1 cacgggtgct tgcacctggt ggcgagtggc gaacgggtga gtaatacatc ggaacatgtc 60 ctgtagtggg ggatagcccg gcgaaagccg gattaatacc gcatacgatc tatggatgaa 120 agcgggggac cttcgggcct cgcgctatag ggttggccga tggctgatta gctagttggt 180 ggggtaaagg cctaccaagg cgacgatcag tagctggtct gagaggacga ccagccacac 240 tgggactgag acacggccca gactcctacg ggaggcagca gtggggaatt ttggacaatg 300 ggcgaaagcc tgatccagca atgccgcgtg tgtgaagaag gccttcgggt tgtaaagcac 360 ttttgtccgg aaagaaatcc ttggctctaa tacagtcggg ggatgacggt accggaagaa 420 taagcaccgg ctaactacgt gccagcagcc gcggtaatac gtagggtgca agcgttaatc 480 ggaattactg ggcgtaaagc gtgcgcaggc ggtttgctaa gaccgatggg aaatccccgg 540 gctcaacctg ggaactgcat tggtgactgg caggctagag tatggcagag gggggtagaa 600 ttccacgtgt agcagtgaaa tgcgtagaga tgtggaggaa taccgatggc gaaggcagcc 660 ccctgggcca atactgacgc tcatgcacga aagcgtgggg agcaaacagg attagatacc 720 ctggtagtcc acgccctaaa cgatgtcaac tagttgttgg ggattcattt ccttagtaac 780 gtagctaacg cgtgaagttg accgcctggg gagtacggtc gcaagattaa aactcaaagg 840 aattgacggg gacccgcaca agcggtggat gatgtggatt aattcgatgc aacgcgaaaa 900 accttaccta cccttgacat ggtcggaatc ctgaagagat tcgggagtgc tcgaaagaga 960 accggcgcac aggtgctgca tggctgtcgt cagctcgtgt cgtgagatgt tgggttaagt 1020 cccgcaacga gcgcaaccct tgtccttagt tgctacgcaa gagcactcta aggagactgc 1080 cggtgacaaa ccggaggaag gtggggatga cgtcaagtcc tcatggccct tatgggtagg 1140 gcttcacacg tcatacaatg gtcggaacag agggttgcca acccgcgagg gggagctaat 1200 cccagaaaac cgatcgtagt ccggattgca ctctgcaact cgagtgcatg aagctggaat 1260 cgctagtaat cgcggatcag catgccgcgg tgaatacgtt cccgggtctt gtacacaccg 1320 cccgtcacac catggggagtg ggttttacca gaagtggcta gtct 1364
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