KR101948233B1 - Compositions for Preventing Hair Loss and Stimulating Hair Sprouting - Google Patents
Compositions for Preventing Hair Loss and Stimulating Hair Sprouting Download PDFInfo
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Abstract
Description
본 발명은 탈모방지 및 발모촉진 조성물에 관한 것이다. The present invention relates to hair loss prevention and hair growth promoting compositions.
두피로부터 모발이 빠지는 탈모증의 원인에는 유전적인 체질이나 남성 호르몬의 작용과 같은 내적인 요인, 일상 생활에서의 정신적인 스트레스, 두피에서의 과산화지질의 축적과 같은 외적인 요인, 및 이들 요인들이 복합적으로 관여되는 요인들이 알려져 있다. 최근에는 남성형 탈모뿐만 아니라, 여성들의 비만성 탈모를 비롯하여 젊은 층에서도 탈모가 점차 확산되는 추세에 있다. 따라서, 이러한 탈모현상을 개선하기 위해 많은 종류의 탈모방지 및 발모촉진제가 개발 및 시판되고 있다.The cause of hair loss from the scalp is caused by internal factors such as genetic constitution, male hormone action, mental stress in daily life, extrinsic factors such as accumulation of lipid peroxidation in the scalp, Are known. In recent years, not only male-type hair loss, but also hair loss in women, such as obesity hair loss is gradually spreading. Accordingly, many types of hair loss prevention and hair growth promoting agents have been developed and marketed to improve such hair loss phenomenon.
가장 일반적으로 알려져 있는 탈모증상 개선을 위한 메커니즘은 남성호르몬을 제어함으로써 탈모를 막고 발모를 촉진시키는 기전이다. 그러나 이와 관련된 대부분의 공지기술들은 그 효과가 미미하거나 부작용이 심하여 안전하고 효율적인으로 탈모방지 및 발모촉진제로 기능하고 있지 못한 실정이다.The most commonly known mechanism for improving hair loss symptoms is the mechanism by which male hormones are controlled to prevent hair loss and promote hair growth. However, most of the known technologies related thereto are not effective as hair loss prevention and hair growth promoting agent because they are insignificant or have a serious side effect and are safe and effective.
한편, 천연추출물을 포함하는 탈모방지 및 발모촉진제도 다양하게 시판되고 있다. 이러한 약물들은 천연추출물에 의한 혈행개선 및 혈관확장에 의해 두피의 혈액순환을 개선하고, 그에 따라 탈모증상을 개선하는 치료 기전을 갖는다. 그러나, 이러한 약물들은 효과가 미미하여 만족할만한 탈모증상 개선 효과를 제공하지 못하고 있는 실정이다. On the other hand, a variety of hair loss prevention and hair growth promoting agents including natural extracts are commercially available. These medicines have a therapeutic mechanism for improving blood circulation of the scalp by improving blood circulation and vasodilation by natural extracts, thereby improving hair loss symptoms. However, these drugs have little effect and thus fail to provide a satisfactory effect of improving hair loss symptoms.
최근에는 TGF-β로 인하여 탈모의 현상이 급격히 이루어지는 것이 보고된 바 있다. 따라서 TGF-β의 억제를 유도함으로 정상 모발이 탈모로 진행되는 과정을 줄여 결과적으로 탈모 현상을 늦추는 탈모증상 개선 약물도 개발되고 있다. Recently, it has been reported that the phenomenon of hair loss due to TGF-β is rapidly performed. Therefore, there is also developed a drug for alleviation of hair loss, which reduces the process of normal hair to progress to hair loss as a result of inducing inhibition of TGF-beta and consequently slows the hair loss phenomenon.
대한민국 등록특허 제10-1841042호는 하기 화학식 1로 표시되는 화합물 또는 이의 약학적으로 허용가능한 염을 유효성분으로 함유하는, 사이클로포스파마이드(cyclophosphamide)의 부작용인 탈모를 개선하거나 억제하는 약학조성물에 관하여 개시하고 있다:Korean Patent No. 10-1841042 discloses a pharmaceutical composition for improving or inhibiting hair loss, which is a side effect of cyclophosphamide, comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient Lt; / RTI >
[화학식 1][Chemical Formula 1]
그러나, 상기 약학조성물은 사이클로포스파마이드(cyclophosphamide)의 부작용에 의한 탈모증상개선과 별개로, 일반적인 탈모증상 개선에는 효과가 충분하지 않은 단점이 있다. 따라서, 일반적인 탈모증상에 사용될 수 있는 효과적인 약물의 개발이 요구되고 있다. However, the pharmaceutical composition has disadvantages in that it is not effective in improving general hair loss symptoms, apart from improvement of hair loss symptoms due to side effects of cyclophosphamide. Therefore, there is a need for the development of effective drugs that can be used for general hair loss symptoms.
본 발명은, 상기와 같은 종래기술의 과제를 해결하기 위하여 안출된 것으로서, 일반적인 탈모증상개선에 효과가 우수하며 부작용으로부터 자유로운 탈모방지 및 발모촉진 조성물을 제공하는 것을 목적으로 한다. Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made keeping in mind the above problems occurring in the prior art, and an object of the present invention is to provide a composition for preventing hair loss and promoting hair growth, which is excellent in improving general hair loss symptoms and free from side effects.
본 발명은, 상기 목적을 달성하기 위하여, The present invention, in order to achieve the above object,
조성물 총 중량에 대하여, 하기 화학식 1로 표시되는 화합물 또는 이의 약학적으로 허용가능한 염 20~50 중량%, L-아르기닌 10~40 중량%, 및 구리 트리펩타이드-1(GHK-Cu) 10~40 중량%를 포함하는 탈모방지 및 발모촉진 조성물을 제공한다: Wherein the composition comprises 20 to 50% by weight of a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof, 10 to 40% by weight of L-arginine, 10 to 40% by weight of copper tripeptide-1 % ≪ / RTI > by weight of a composition for promoting hair loss prevention and hair growth:
[화학식 1][Chemical Formula 1]
상기 화학식 1에서, R1 및 R2는 각각 동일하거나 다를 수 있으며, 수소, (C1~C4)알킬, (C1~C4)알콕시 및 할로겐으로 이루어진 군에서 선택될 수 있다.In Formula 1, R 1 and R 2 may be the same or different and may be selected from the group consisting of hydrogen, (C 1 -C 4) alkyl, (C 1 -C 4) alkoxy and halogen.
본 발명의 일실시형태에서, 상기 조성물은 조성물 총 중량에 대하여, 풀루란 5~20 중량%; 콘알부민 5~20 중량%; 보스웰릭산 5~20 중량%; 및 수염가래꽃 및 산두근을 0.5~1:0.5~1의 중량비로 혼합한 혼합물의 추출물 10~20 중량%를 더 포함할 수 있다. In one embodiment of the invention, the composition comprises 5-20% by weight of pullulan, based on the total weight of the composition; 5 to 20% by weight of corn albumin; 5 to 20% by weight of boswelly acid; And 10 to 20% by weight of an extract of a mixture of a beard sprout flower and an acid beet pulp at a weight ratio of 0.5 to 1: 0.5 to 1.
본 발명의 일실시형태에서, 상기 조성물에 있어서 수염가래꽃 및 산두근을 0.5:0.5:1의 중량비로 혼합한 혼합물의 추출물은 추출용매로 에탄올을 사용하여 에탄올의 끓는 온도에서 0.3~1.5 시간 추출한 추출물일 수 있다. In one embodiment of the present invention, the extract of the mixture obtained by mixing the whitish spore flower and the acidic beef at a weight ratio of 0.5: 0.5: 1 is extracted with 0.3 to 1.5 hours at the boiling temperature of ethanol using ethanol as an extraction solvent Lt; / RTI >
본 발명의 일실시형태에서, 상기 탈모방지 및 발모촉진 조성물은 화장료 조성물로서 샴프 또는 헤어토닉 등일 수 있다. In one embodiment of the present invention, the composition for promoting hair loss prevention and hair growth may be a shampoo, a hair tonic, or the like as a cosmetic composition.
본 발명의 탈모방지 및 발모촉진 조성물은 부작용 없이 일반적인 탈모증상개선에 매우 우수한 효과를 제공한다. The hair loss prevention and hair growth promoting composition of the present invention provides an excellent effect for improving general hair loss symptoms without side effects.
도 1은 마우스 경피 투여에 의한 확인된 본 발명의 탈모방지 및 발모촉진 조성물의 모발 재성장 효과를 나타낸 사진이다.
도 2 및 도 3은 본 발명의 탈모방지 및 발모촉진 조성물로서 헤어토닉을 15주 동안 적용한 경우의 탈모방지개선 효과를 나타낸 사진이다.BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a photograph showing the hair regrowth preventing effect of the hair loss prevention and hair growth promoting composition of the present invention confirmed by transdermal administration of a mouse.
FIG. 2 and FIG. 3 are photographs showing the effect of improving hair loss prevention when hair tonic is applied for 15 weeks as a hair loss prevention and hair growth promoting composition of the present invention.
이하에서 본 발명을 바람직한 실시예를 참고하여 설명한다. Hereinafter, the present invention will be described with reference to preferred embodiments.
본 발명은, According to the present invention,
조성물 총 중량에 대하여, 하기 화학식 1로 표시되는 화합물 또는 이의 약학적으로 허용가능한 염 20~50 중량%, L-아르기닌 10~40 중량%, 및 구리 트리펩타이드-1(GHK-Cu) 10~40 중량%를 포함하는 탈모방지 및 발모촉진 조성물에 관한 것이다: Wherein the composition comprises 20 to 50% by weight of a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof, 10 to 40% by weight of L-arginine, 10 to 40% by weight of copper tripeptide-1 % ≪ / RTI > by weight of a composition for promoting hair loss and promoting hair growth,
[화학식 1][Chemical Formula 1]
상기 화학식 1에서, R1 및 R2는 각각 동일하거나 다를 수 있으며, 수소, (C1~C4)알킬, (C1~C4)알콕시 및 할로겐으로 이루어진 군에서 선택될 수 있다.In Formula 1, R 1 and R 2 may be the same or different and may be selected from the group consisting of hydrogen, (C 1 -C 4) alkyl, (C 1 -C 4) alkoxy and halogen.
상기 화학식 1의 화합물은 사이클로포스파마이드(cyclophosphamide)의 부작용인 탈모를 개선하는 효과가 있는 것으로 알려져 있다. 그러나, 일반적인 탈모증상개선에는 그 효과가 충분하지 못한 단점이 있었다.The compound of Formula 1 is known to have the effect of improving hair loss, which is a side effect of cyclophosphamide. However, there is a disadvantage in that the effect of improving general hair loss symptoms is not sufficient.
따라서, 본 발명자들은 상기 화학식 1의 화합물과 상승효과를 나타낼 수 있는 추가의 성분들을 조합하여 본 발명의 조성물을 완성하였다.Therefore, the present inventors completed the composition of the present invention by combining the above-mentioned compound of Formula 1 with additional components capable of exhibiting a synergistic effect.
본 발명의 일실시형태에서, 상기 조성물은 조성물 총 중량에 대하여, 풀루란 5~20 중량%; 콘알부민 5~20 중량%; 보스웰릭산 5~20 중량%; 및 수염가래꽃 및 산두근을 0.5~1:0.5~1의 중량비로 혼합한 혼합물의 추출물 10~20 중량%를 더 포함할 수 있다. In one embodiment of the invention, the composition comprises 5-20% by weight of pullulan, based on the total weight of the composition; 5 to 20% by weight of corn albumin; 5 to 20% by weight of boswelly acid; And 10 to 20% by weight of an extract of a mixture of a beard sprout flower and an acid beet pulp at a weight ratio of 0.5 to 1: 0.5 to 1.
상기 풀루란(exopolysaccharide, EPS pullulan)은 곰팡이에 의한 세포외 분비 다당류로서, 1938년 Bauer에 의해 처음 발견된 후 많은 연구에 의해 안전한 중성용액으로 알려져 있다. The exopolysaccharide (EPS pullulan) is an extracellular secretory polysaccharide produced by the fungus. It was first discovered by Bauer in 1938 and is known as a safe solution by many studies.
풀루란은 maltotriose 단위가 α-1, 6 결합으로 연결된 것으로 진균류의 일종인 Aureobasidium pullulans가 생성하는 수용성 다당류이다. 버섯의 자실체에는 β-1, 3 glycoside 결합을 주쇄로 하고 그 C-6 위치에 1개의 glucose 잔기가 붙어 있는 구조를 가진 다당류가 함유되어 있다. 최근 이러한 구조를 가지는 β-glucan 중에는 종양세포의 증식을 저지하는 작용이 있는 것이 밝혀져 주목되고 있다. Pullulan is a water-soluble polysaccharide produced by the fungus Aureobasidium pullulans, which is linked to maltotriose units by α-1, 6 bonds. Fruiting bodies of mushrooms contain polysaccharides having a structure with β-1, 3 glycoside bonds as the main chain and a glucose residue attached at its C-6 position. Recently, β-glucan having such a structure has been found to have an action of inhibiting the proliferation of tumor cells, and attention has been paid.
상기 콘알부민은 오보트랜스페린(Ovotransferrin)이라고도 불리며, 난백 단백질의 12.6%를 차지하는 철이 결합된 분자량 76.6kDa의 당단백질로서 등전점(pI)은 6.1이다. 난백 단백질에서 두 번째로 많은 콘알부민은 686개의 아미노산 잔기를 가지고 있으며, 15개의 이황화 결합과 4개의 만노오스(mannose) 그리고 8개의 노르말-아셀틸글루코사민(N-acetylglucosamin)으로 구성된 한 개의 글리칸(glycan) 사슬로 당화되어 있다. 콘알부민은 두개의 금속이온(Fe3+, Al3+, Cu2+, Zn2+ 등)이 pH 6에서 열에 안정한 복합체를 형성하고 있다. 이들 복합체들은 각각, 붉은색, 무색, 노란색, 무색을 띈다. 콘알부민은 Fe3+원자가 하나도 없는 것, 한 개 또는 두개를 갖고 있는 것의 3가지 형이 균형 있게 존재하며, 이러한 철 결합능은 지질산화를 방지하는데 간접적인 역할을 한다. 또한, 금속이온과의 결합으로 열변성과 미생물오염에 대해 안전하고 철결합 복합체와 비철결합 아포단백질간의 다양한 특성으로 난백의 가공과 기능적 특성에 중요한 역할을 한다.The above-mentioned conalumin is also called ovotransferrin, which is a glycoprotein having a molecular weight of 76.6 kDa which is iron-bonded and occupies 12.6% of the egg white protein. The isoelectric point (pI) is 6.1. The second largest conglomerate of egg white protein has 686 amino acid residues and is composed of one glycan (N-acetylglucosamine) consisting of 15 disulfide bonds, 4 mannose and 8 N-acetylglucosamine ) Chain. Conalbumin is a thermally stable complex at pH 6 with two metal ions (Fe3 +, Al3 +, Cu2 +, Zn2 +, etc.). These complexes are reddish, colorless, yellow, and colorless, respectively. There are three types of conal albumin, one without Fe 3+ atom and one with two or more, and this iron binding ability plays an indirect role in preventing lipid oxidation. In addition, it is safe for thermal deformation and microbial contamination by bonding with metal ions, and plays an important role in processing and functional characteristics of egg white with various characteristics between iron-binding complex and non-iron-binding apoprotein.
상기 보스웰릭산은 보스웰리아 세레타 나무의 줄기에 상처를 내면 흘러나오는 엷은 녹색을 띤 진액으로부터 얻어진다. 보스웰리아 세레타는 아프리카, 남부 아라비아 또는 인도 등지에서 자생한다. 보스웰릭산은 전통적으로 피부 발진 및 궤양 등의 염증이나 천식, 기관지염 및 후두염과 같은 호흡기 질환에 사용되었고, 관절염 등에도 의학적 효과가 있는 것으로 알려져 있다. The boswellic acid is obtained from the pale greenish juice which flows when the boswellia serrata tree is injured. Bosswellia Sereta is native to Africa, southern Arabia or India. Bosswelllic acid has traditionally been used for inflammation such as skin rashes and ulcers, respiratory diseases such as asthma, bronchitis and laryngitis, and it has been known to have medical effects such as arthritis.
보스웰리산은 우수한 엘라스타아제 활성 저해 및 글리코스아미노글리칸 분해 억제 효과를 나타내는 것으로 알려져 있다.Bosswelli acid is known to exhibit excellent inhibition of elastase activity and inhibition of glycosaminoglycan degradation.
상기 수염가래꽃 및 산두근을 0.5~1:0.5~1의 중량비로 혼합한 혼합물의 추출물은 추출용매로 에탄올을 사용하여 에탄올의 끓는 온도에서 0.3~1.5 시간 추출한 추출물이 바람직하게 사용될 수 있다. The extract of the mixture obtained by mixing the above-mentioned whitish whiting flowers and the acidic beef at a weight ratio of 0.5: 1: 0.5 to 1 may be preferably used as an extraction solvent and extracted with ethanol at 0.3 to 1.5 hours at the boiling temperature of ethanol.
상기 수염가래꽃은 한의학에서 생약으로 사용되고 있는 약용식물로서 다양한 생리활성물질을 함유하고 있다. 수염가래꽃 분말은 반변련, 반변란 또는 급해색이라고 불리며 국내에서는 중부 이남과 제주도에 분포하고 도랑이나 냇가, 논두렁의 습지에서 서식하고 있다. 수염가래꽃 분말은 전통적으로 이뇨, 소염, 해독 등의 효능을 가지고 있으며 천식, 호흡곤란, 백일해, 악성종기, 습진, 외상출혈을 치료하는 약재로 사용되었다.The beard sprout flower is a medicinal plant used as a herbal medicine in Oriental medicine and contains various physiologically active substances. The beard spore flower powder is called semi-mature, semi-irregular or rapid-hueing color. It is distributed in the southern part of central Korea and Jeju-do, and it lives in ditches, streams and wetlands of rice paddies. Beard flies have traditionally been used as medicines for the treatment of asthma, dyspnea, pertussis, malignant swelling, eczema and traumatic hemorrhage.
상기 산두근은 새모래덩굴(Menispermum dahuricum DC.)로도 불리우는 덩굴뻗는 떨기나무 뿌리이다. 우리나라 전국 각지 양지바른 산기슭에 서식하며 뿌리와 줄기에 알카로이드, 타우리신, 테르난드린 등이 함유되어 있는 것으로 알려져 있다(Yakugawa Zasshi 1970). 또한, 시노메닌, 타우리신은 진경작용이 있으며 고혈압 및 항염증 작용에도 효능이 있는 것으로 알려져 있으며, 현재 총 알카로이드를 추출 분류하여 류마치즘 치료제로의 접근이 연구되어 지고 있다(Acta Med Okayama 1976).The mountain beetle is a vine extending bush root, also called a new sand vine (Menispermum dahuricum DC.). It is known to contain alkaloids, taurine, and terenadrine in roots and stems (Yakugawa Zasshi 1970). In addition, synomenin and taurine are known to have antipruritic effects and are also effective against hypertension and anti-inflammatory actions. Acta Med Okayama (1976) .
본 발명의 일실시형태에서, 상기 탈모방지 및 발모촉진 조성물은 화장료 조성물로서 샴프 또는 헤어토닉 등일 수 있다. In one embodiment of the present invention, the composition for promoting hair loss prevention and hair growth may be a shampoo, a hair tonic, or the like as a cosmetic composition.
이하에서, 실시예를 통하여 본 발명을 보다 상세히 설명한다. 그러나, 하기의 실시예는 본 발명을 더욱 구체적으로 설명하기 위한 것으로서, 본 발명의 범위가 하기의 실시예에 의하여 한정되는 것은 아니다. 하기의 실시예는 본 발명의 범위 내에서 당업자에 의해 적절히 수정, 변경될 수 있다. Hereinafter, the present invention will be described in more detail by way of examples. However, the following examples are intended to further illustrate the present invention, and the scope of the present invention is not limited by the following examples. The following examples can be appropriately modified and changed by those skilled in the art within the scope of the present invention.
제조예 1: 수염가래꽃 및 산두근 추출물 제조Production Example 1: Production of beard sprout flower and mountain beet roots extract
건조된 수염가래꽃과 산두근 각 500g씩을 가열용기에 넣고, 에탄올 2L를 추출용매로 넣어 끓는 온도에서 30분간 가열하여 수염가래꽃과 산두근 추출물을 제조하였다.500 g each of dried beard spiny flowers and mountain hulls were placed in a heating vessel, and 2 L of ethanol was added as an extraction solvent and heated at a boiling temperature for 30 minutes to prepare a beard spelea flower and an extract of beetroot.
실시예 1: 탈모방지 및 발모촉진 조성물의 제조Example 1: Prevention of hair loss and preparation of hair growth promoting composition
화학식 1의 화합물의 약제학적으로 허용가능한 염으로서 Sigma사에서 제조한 SML0128 40 중량%, L-아르기닌 30 중량% 및 구리 트리펩타이드-1(GHK-Cu) 30 중량%를 혼합하여 탈모방지 및 발모촉진 조성물을 제조하였다.40% by weight of SML0128 manufactured by Sigma, 30% by weight of L-arginine and 30% by weight of copper tripeptide-1 (GHK-Cu) as a pharmaceutically acceptable salt of the compound of the formula (1) A composition was prepared.
실시예 2: 탈모방지 및 발모촉진 조성물의 제조Example 2: Preparation of hair loss prevention and hair growth promoting composition
화학식 1의 화합물의 약제학적으로 허용가능한 염으로서 Sigma사에서 제조한 SML0128 40 중량%, L-아르기닌 15 중량%, 구리 트리펩타이드-1(GHK-Cu) 20 중량%, 풀루란 5 중량%, 콘알부민 5 중량%, 보스웰릭산 10 중량%, 및 상기 제조예 1에서 제조한 수염가래꽃 및 산두근 혼합물의 추출물 5 중량를 혼합하여 탈모방지 및 발모촉진 조성물을 제조하였다.40% by weight of SML0128 manufactured by Sigma, 15% by weight of L-arginine, 20% by weight of copper tripeptide-1 (GHK-Cu), 5% by weight of pullulan, 5% by weight of albumin, 10% by weight of boswellylic acid, and 5% by weight of the extract of the whiskers and balsam mixture prepared in Preparation Example 1 were mixed to prepare a composition for promoting hair loss prevention and hair growth.
비교예 1:Comparative Example 1: 탈모방지 및 발모촉진제 조성물의 제조Preparation of Hair Loss Prevention and Hair Growth Promoter Composition
화학식 1의 화합물의 약제학적으로 허용가능한 염으로서 Sigma사에서 제조한 SML0128 100%로 탈모방지 및 발모촉진제를 제조하였다.As a pharmaceutically acceptable salt of the compound of the formula (1), hair loss preventing and hair growth promoting agent was prepared by 100% of SML0128 manufactured by Sigma.
<Sigma사에서 제조한 SML0128>≪ SML0128 manufactured by Sigma &
실시예 3~4 및 비교예 2: 탈모방지 및 발모촉진 조성물을 포함하는 헤어토닉 제조Examples 3 to 4 and Comparative Example 2: Hairtonic production comprising hair loss prevention and hair growth promoting composition
하기 표 1의 조성에 따라, 헤어토닉 조성물을 제조하였다. A hair tonic composition was prepared according to the composition of Table 1 below.
(실시예 1 제조)30
(Preparation of Example 1)
(실시예 2 제조)30
(Preparation of Example 2)
(비교예 1 제조)30
(Preparation of Comparative Example 1)
시험예: 발모 촉진 또는 탈모 억제 효과 실험Test Example: Effect of stimulation of hair growth or hair loss inhibition
시험예 1: 사람Test Example 1: 모유두세포 증식 효과 평가Evaluation of proliferation of dermal papilla cells
상기 실시예 1 및 실시예 2의 조성물, 및 비교예 1의 시료가 사람 모유두세포(human hair dermal papilla cell)의 세포 증식에 미치는 효과를 아래와 같이 사람 모유두세포(human hair dermal papilla cell, HHDPC)(Sciencell사, cat. no. 2400)을 사용하여 평가하였다.The effects of the compositions of Examples 1 and 2 and the sample of Comparative Example 1 on the cell proliferation of human hair dermal papilla cells were examined by using human hair dermal papilla cells (HHDPC) Sciencell, cat. No. 2400).
10% FBS (fetal bovine serum, Cambrex)가 포함된 전용배지인 MSC(Mesenchymal Stem Cell) 배지에 사람 모유두 세포(Sciencell사, cat. no. 2400)를 0.5 × 104cells이 되도록 하여 96 웰 플레이트에 접종하여 24 시간 동안, 37℃, 5% CO2 습윤 조건 하에서 배양하였다. 상기 배양액에 상기 시료들을 10 ㎍/ml의 농도로 처리하여 72 시간 동안 배양한 후, 5 mg의 MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) 시약 20㎕를 상기 배양액에 넣고 37℃, 5% CO2 습윤 조건하에서 4시간 동안 배양하였다. 배양 시, 살아있는 세포에는 보라색의 크리스탈이 형성된다. 4시간 배양 후 배지를 제거하고 DMSO 100를 넣어 10분 동안 쉐이킹 인큐베이터(shaking incubator)에서 크리스탈을 녹인 후 570 nm에서 흡광도를 측정하여 세포 성장 정도를 측정하였다. 상기 시험결과를 대조군인 무처리군 대비 백분율로 아래 표 2에 나타내었다.The cells were plated on a 96-well plate so that human dermal papilla cells (Sciencell, cat. No. 2400) were cultured at 0.5 × 10 4 cells in a MSC (Mesenchymal Stem Cell) medium, a culture medium containing 10% fetal bovine serum inoculation and for 24 hours and then cultured under 37 ℃, 5% CO 2 wet condition. The culture was treated with the above samples at a concentration of 10 μg / ml and cultured for 72 hours. 5 mg of MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide) Insert the 20㎕ the culture was incubated for 4 hours in a 37 ℃, 5% CO 2 wet condition. In culture, purple crystals form in living cells. After 4 hours of incubation, the medium was removed, DMSO 100 was added, the crystal was dissolved in a shaking incubator for 10 minutes, and the absorbance was measured at 570 nm to measure the degree of cell growth. The test results are shown in Table 2 below as a percentage of the control group as the non-treatment group.
상기 표 2의 결과에 의하면, 본 발명의 실시예들의 시료는 대조군 및 비교예 1의 조성물과 비교하여 현저한 모유두세포의 증식을 촉진하는 것으로 확인되었다. 특히, 실시예 2의 조성물은 더 우수하게 모유두세포의 증식을 촉진하는 것으로 확인되었다.The results of Table 2 above confirm that the samples of the examples of the present invention promote remarkable proliferation of dermal papilla cells compared with the control and the composition of Comparative Example 1. [ In particular, the composition of Example 2 was found to better promote the proliferation of dermal papilla cells.
시험예 2: 사람 각질형성세포(HaCaT cell) 증식 효과 평가Test Example 2: Evaluation of proliferation effect of human keratinocyte (HaCaT cell)
상기 실시예 1 및 실시예 2의 조성물, 및 비교예 1의 시료가 사람 각질형성세포의 세포 성장에 미치는 효과를, 각질형성세포(HaCaT cell)(Dr. N. E. Fusenig (Heidelberg, Germany)을 사용하여, 상기 시험예 1과 동일한 방법으로 평가하였다. 배지는 10% FBS(fetal bovine serum, Cambrex)가 포함된 RPMI 배지를 사용하였다. 시험결과를 대조군인 무처리군 대비 백분율로 아래 표 3에 나타내었다.The effect of the compositions of Examples 1 and 2 and the sample of Comparative Example 1 on the cell growth of human keratinocytes was examined using keratinocytes (HaCaT cell) (Dr. NE Fusenig (Heidelberg, Germany) , And evaluated in the same manner as in Test Example 1. The RPMI medium containing 10% FBS (fetal bovine serum, Cambrex) was used as the medium, and the results are shown in Table 3 below as a percentage of the control group .
상기 표 3의 결과에 의하면, 본 발명의 실시예들의 시료는 대조군 및 비교예 1의 조성물과 비교하여 현저한 각질형성세포의 증식을 촉진하는 것으로 확인되었다. 특히, 실시예 2의 조성물은 더 우수하게 각질형성세포의 증식을 촉진하는 것으로 확인되었다.According to the results shown in Table 3, the samples of the examples of the present invention were found to promote proliferation of keratinocytes remarkably as compared with the control and the composition of Comparative Example 1. In particular, the composition of Example 2 was found to promote the proliferation of keratinocytes to be superior.
시험예 3: 모발 성장효과에 대한 동물 실험Test Example 3: Animal experiment for hair growth effect
실시예 1 및 실시예 2의 조성물, 및 비교예 1의 시료의 탈모 증상 개선 효과를 확인하기 위해, 동물모델로 많이 사용하는 자발적 탈모의 유전적 특성을 가진 6주령의 암컷 C57BL/6 마우스를 이용하였다. 암컷 C57BL/6 마우스를 대상으로 일렉트릭 클립퍼 커트(electric clipper cut)를 이용하여 1차 제모 후 쉐이빙 폼(shaving form)을 바르고, 면도날(shaving razor)을 이용하여 가로로 등쪽 경부에서 꼬리 기시부까지의 길이와 세로로 등쪽 정중선을 기준으로 좌우 각각 1cm 이상의 폭에 대하여 2차 제모를 실시하여 등쪽 피부색이 균질한 핑크색을 보이는 개체만을 선별하여 실험에 사용하였다. 제모 부위에 붓을 댄 후 시험물질 200㎕를 마이크로피펫(micropipette)을 이용하여 붓끝에 분주한 후 상하 방향으로 제모 부위 전체에 균질하게 1일 1회 21일간 도포하였다. 모든 실험 결과는 평균치±표준편차로 표시하고 모든 데이터에 대해 분산의 동질성 비교분석은 터키 검증(Turkey test) 및 ANOVA(one-way analysis of variance)를 실시하였으며, 대조군과의 유의적 차이가 있는 실험군에 대해 두네트 티 검증(Dunnett's t-test)을 실시하였다.6-week-old female C57BL / 6 mice with the genetic characteristic of spontaneous hair loss, which is frequently used as an animal model, was used to examine the effect of the compositions of Examples 1 and 2 and the sample of Comparative Example 1 on improving hair loss symptoms Respectively. Female C57BL / 6 mice were subjected to primary shaving using an electric clipper cut, followed by shaving with a shaving razor. The shaving razor was applied to the cervical vertebra The length and length of the midline of the dorsum were compared with the width of 1cm or more on each side of the median line, and only the individuals showing a homogeneous pink color of the dorsal skin were selected for the experiment. After applying a brush to the hair removal part, 200 쨉 L of the test material was dispensed on the end of the brush using a micropipette, and then uniformly applied to the entire hair removal part once a day for 21 days in the vertical direction. All test results were expressed as mean ± standard deviation. For all data, homogeneity analysis of variance was performed with Turkey test and ANOVA (one way analysis of variance) (Dunnett's t-test).
시험예 3-1: 양모면적 분석Test Example 3-1: Analysis of wool area
실시예 1 및 실시예 2의 조성물, 및 비교예 1의 시료를 암컷 C57BL/6 마우스에 경피 투여한 후 0, 7, 14 및 21일째 총 4회에 걸쳐 경시적인 모발 성장 주기(hair growth cycle)의 변화를 육안으로 확인하였다. 그 결과, 하기 표 4 및 도 1에 개시한 바와 같이 초기 7일부터 털이 꼬리 쪽 등 부위와 목 쪽 등 부위로 모발 성장을 나타내었다. 14일부터 아무것도 처리하지 않은 대조군에 대비하여 비교예 1인 화학식 1의 화합물의 염(Sigma사에서 제조한 SML0128), 실시예 1의 조성물 및 실시예 2의 조성물을 처리하였을 때, 더 우수한 모발 재성장 효과를 나타냈다. The compositions of Example 1 and Example 2 and the sample of Comparative Example 1 were transdermally administered to female C57BL / 6 mice, and then subjected to a hair growth cycle over a total of 4 times on days 0, 7, 14 and 21, Were visually confirmed. As a result, as shown in Table 4 and Fig. 1, hair growth was observed from the beginning 7 days to hairy tail side back part and neck side back part. Treatment of the salt of the compound of formula 1 (SML0128 produced by Sigma), the composition of Example 1 and the composition of Example 2 in comparison to the control group that did not undergo anything from Day 14 resulted in better hair regrowth Effect.
더 나아가서, 비교예 1인 화학식 1의 화합물의 염(Sigma사에서 제조한 SML0128)과 비교하여, 실시예 1의 조성물 및 실시예 2의 조성물은 더 현저한 모발 재성장 효과를 나타냈다. 특별히, 실시예 2의 조성물은 실시예 1보다도 더 현저한 모발 재성장 효과를 나타냈다. Furthermore, the composition of Example 1 and the composition of Example 2 exhibited a more remarkable hair regrowth effect, as compared to the salt of the compound of Formula 1 (SML0128 produced by Sigma). Specifically, the composition of Example 2 exhibited a hair regrowth effect more prominent than that of Example 1.
**탈모증 유도 C57BL/6 마우스에 대한 시료 처리에 의한 모발 재성장 촉진 효과** Promotes hair re-growth by sample treatment of alopecia induced C57BL / 6 mice
시험예 3-2: 조직병리학적 분석Test Example 3-2: Histopathological Analysis
암컷 C57BL/6 마우스 모낭의 형태학적인 변화를 확인하기 위해, 경피투여 21일째에 채취한 피부조직을 헤마톡실린-에오신(hematoxylin-eosin)으로 염색하여 광학현미경으로 관찰하여 모낭의 수를 확인하고, 그 결과를 하기 표 5에 나타내었다. In order to confirm the morphological changes of female C57BL / 6 mouse hair follicles, the skin tissue collected on the 21st day after transdermal administration was stained with hematoxylin-eosin and observed with an optical microscope to confirm the number of hair follicles, The results are shown in Table 5 below.
**탈모증 유도 C57BL/6 마우스에서 시료 처리에 따른 휴지기 모낭의 수 측정 결과상기 표 5 나타낸 바와 같이, 비교예 1인 화학식 1의 화합물의 염(Sigma사에서 제조한 SML0128)으로 처리한 경우, 대조군과 비교하여 휴지기의 모낭수가 증가한 것을 확인할 수 있다. ** Results of measurement of number of resting hair follicles following sample treatment in alopecia induced C57BL / 6 mice As shown in Table 5, when treated with the salt of the compound of Chemical Formula 1 (SML0128 produced by Sigma), the control group The number of hair follicles in the resting period was increased.
그러나, 본 발명의 실시예 1의 조성물로 처리한 경우, 대조군이나 비교예 1과 비교하여 휴지기의 모낭수가 현저하게 증가한 것을 확인할 수 있다. However, when treated with the composition of Example 1 of the present invention, it was confirmed that the number of hair follicles at rest was significantly increased as compared with the control group and Comparative Example 1. [
특히, 본 발명의 실시예 2의 조성물로 처리한 경우, 실시예 1과 비교해서도 휴지기의 모낭수가 현저하게 증가한 것을 확인할 수 있다. In particular, when treated with the composition of Example 2 of the present invention, the number of hair follicles in the resting period was significantly increased as compared with Example 1.
시험예 4: 탈모방지 및 발모촉진 효과에 대한 임상시험Test Example 4: Clinical trial for prevention of hair loss and promotion of hair growth
본 실시예 4, 실시예 5 및 비교예 2의 헤어토닉 조성물을 100 ml를 스프레이 용기에 넣어 1일 2회(아침, 저녁) 탈모증 환자(각 실험군 마다 30~50 대 남자 8명, 여자 2명)의 탈모부위의 두피에 스프레이하고 그 효과를 확인하였다. 상기 시험은 15주 동안 진행하였다. 100 ml of the hair tonic composition of Example 4, Example 5 and Comparative Example 2 were put into a spray container and the patients were alopeccated twice a day (morning and evening) (8 male and 2 female ), And the effect was confirmed. The test was conducted for 15 weeks.
상기 탈모증 환자는 병원을 처음 방문했을 때, 모발 상태를 진단하고, 탈모 부위를 표시 기록하고, 가장 심한 부위에 검은 점을 문신하여 측정 시마다 동일부위를 측정할 수 있도록 조치하였다. 그리고, simple hand-held phototricho gram(Folliscope,Lead M Co,Seoul, Korea)을 이용해 탈모부위의 단위면적당 모발수(모발 개수/㎠)를 측정하였다. 단위 면적당 모발 개수는 30배 렌즈를 이용하여 표시부분 주위 탈모 부위를 촬영후 단위면적내 성모 모발(terminal hair)의 개수를 계산하였다. When the patient visited the hospital for the first time, the hair loss was diagnosed, the hair loss area was recorded and the black spot was tattooed on the most severe area so that the same area could be measured every time the hair loss was measured. The number of hairs per unit area (number of hairs / ㎠) of hair loss area was measured using a simple hand-held phototricho gram (Folliscope, Lead M Co., Seoul, Korea). The number of hairs per unit area was measured by using a 30x lens and the number of terminal hair in a unit area was calculated after the hair loss area around the display area was photographed.
상기 탈모증 환자들의 약물 투여전, 약물 투여후 일정 시점에서 측정된 모발수(모발 개수/㎠)의 변화는 하기 표 6에 나타내었다. 실시예 4의 헤어토닉을 사용한 1번 및 7번 피험자의 시작 시점과 15주 시점에서 촬영한 사진을 각각 도 2 및 도 3에 도시하였다. Table 6 shows changes in the number of hairs (number of hairs / cm2) measured at a certain point in time before and after drug administration in the alopecia patients. 2 and 3 are photographs taken at the starting point and the 15th point of time of the subjects 1 and 7 using the hair tonic of Example 4, respectively.
상기 표 6에 나타낸 바와 같이, 비교예 2의 헤어토닉(Sigma사에서 제조한 SML0128 포함)을 적용한 경우와 비교하여, 본 발명의 실시예 3(실시예 1의 조성물 포함) 및 실시예 4(실시예 2의 조성물 포함)의 헤어토닉을 적용한 피험군들의 경우, 증가된 모발 개수가 현저히 증가한 것을 알 수 있다. As shown in Table 6, in comparison with the case where the hair tonic of Comparative Example 2 (including SML0128 manufactured by Sigma) was applied, Example 3 of the present invention (including the composition of Example 1) and Example 4 In the case of the experimental groups to which the hair tonic of the Example 2 was applied, the number of the increased hair was remarkably increased.
특히, 본 발명의 실시예 3의 헤어토닉을 적용한 경우와 비교하여 실시예 4의 헤어토닉을 비교한 경우 증가된 모발 개수가 더 현저하게 증가한 것을 확인할 수 있다. In particular, it can be seen that the number of hair increased was significantly increased when the hair tonic of Example 4 was compared with the hair tonic of Example 3 of the present invention.
시험예 5: 부작용 평가Test Example 5: Evaluation of side effects
상기 시험예 4의 과정에서 사용된 실시예 3, 실시예 4 및 비교예 2의 헤어토닉의 부작용을 설문을 통하여 확인하고, 그 결과를 하기 표 7에 나타내었다. The side effects of the hair tonics of Example 3, Example 4 and Comparative Example 2 used in the process of Test Example 4 were confirmed through questionnaires, and the results are shown in Table 7 below.
상기 표 7에 나타난 바와 같이, 본 발명의 탈모방지 및 발모촉진 조성물은 부작용이 전혀 발현되지 않았으며 치료 포기자도 전혀 발생하지 않아서 매우 안전한 것으로 확인되었다. As shown in Table 7, it was confirmed that the hair loss prevention and hair growth promoting composition of the present invention was completely safe because no side effects were exhibited at all and no treatment abandoner occurred at all.
Claims (4)
상기 수염가래꽃 및 산두근 혼합물의 추출물은 추출용매로 에탄올을 사용하여 에탄올의 끓는 온도에서 0.3~1.5 시간 추출한 추출물인 것을 특징으로 하는 탈모방지 및 발모촉진용 화장료 조성물.
[화학식 1]
상기 화학식 1에서, R1 및 R2는 각각 동일하거나 다를 수 있으며, 수소, (C1~C4)알킬, (C1~C4)알콕시 및 할로겐으로 이루어진 군에서 선택될 수 있다.Wherein the composition comprises 20 to 50% by weight of a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof, 10 to 40% by weight of L-arginine, 10 to 40% by weight of copper tripeptide-1 (GHK- An extract of a mixture obtained by mixing 5 to 20% by weight of pullulan, 5 to 20% by weight of corn albumin, 5 to 20% by weight of boswellinic acid, and 0.5 to 1: 10 to 20% by weight,
The cosmetic composition for preventing hair loss and promoting hair growth according to claim 1, wherein the extract of the whitish phalaenopsis flower and the mixture of persimmon extract is an extract which is extracted from ethanol at boiling temperature for 0.3 to 1.5 hours using ethanol as an extraction solvent.
[Chemical Formula 1]
In Formula 1, R 1 and R 2 may be the same or different and may be selected from the group consisting of hydrogen, (C 1 -C 4) alkyl, (C 1 -C 4) alkoxy and halogen.
상기 화장료는 샴프 또는 헤어토닉인 것을 특징으로 하는 탈모방지 및 발모촉진용 화장료 조성물. The method according to claim 1,
The cosmetic composition for preventing hair loss and promoting hair growth, wherein the cosmetic composition is a shampoo or a hair tonic.
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