KR20180103801A - A pharmaceutical composition comprising phytosphingosine as an active ingredient for treatment of senile dry skin - Google Patents
A pharmaceutical composition comprising phytosphingosine as an active ingredient for treatment of senile dry skin Download PDFInfo
- Publication number
- KR20180103801A KR20180103801A KR1020180107812A KR20180107812A KR20180103801A KR 20180103801 A KR20180103801 A KR 20180103801A KR 1020180107812 A KR1020180107812 A KR 1020180107812A KR 20180107812 A KR20180107812 A KR 20180107812A KR 20180103801 A KR20180103801 A KR 20180103801A
- Authority
- KR
- South Korea
- Prior art keywords
- phytosphingosine
- skin
- expression
- pharmaceutical composition
- active ingredient
- Prior art date
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- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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Abstract
Description
본 발명은 피토스핑고신을 유효성분으로 함유하는, 노인성 건조증 예방 또는 치료용 약학적 조성물에 관한 것이다.The present invention relates to a pharmaceutical composition for preventing or treating senile dryness comprising phytosphingosine as an active ingredient.
평균 수명의 연장, 신생아의 출산저하 등으로 전체 인구에서 노인 인구가 차지하는 비율이 급격히 증가하고 있다. 통계청 보고에 따르면 우리나라는 2000년에 이미 고령화 사회로 진입했고, 2018년이면 65세 이상의 인구가 15%를 넘는 초고령 사회가 될 것으로 전망되고 있다.The proportion of the elderly population in the total population is rapidly increasing due to an increase in the average life expectancy and a decrease in the birth rate of the newborn. According to the National Statistical Office, Korea has already entered an aging society in 2000, and by 2018 it is expected to become a super-aged society with a population of over 65 years old, which is over 15%.
이처럼 고령화 사회가 전개되면서 노인층 피부 질환자도 급속도로 늘고 있다. 노화된 피부는 피부 질환이 잘 낫지 않고 세균 감염 등의 2차 질환의 위험성이 높아 조기에 치료를 받는 것이 중요하다. 하지만 대한피부과학회에 따르면 노인층의 45%가 1가지 이상의 피부질환을 겪고 있지만, 59%는 피부질환 증상을 가볍게 보거나 진료비 부담 등을 이유로 치료를 등한시하고 있는 실정이다.As the aging society develops, the number of people with skin diseases of the elderly is rapidly increasing. It is important to get early treatment because aging skin does not heal skin disease well and there is a high risk of secondary disease such as bacterial infection. However, according to the Korean Dermatological Association, 45% of the elderly suffer from one or more skin diseases, but 59% are neglecting the treatment because of the skin disease symptoms or the burden of medical expenses.
피부노화는 유전적으로 결정되는 내인성 노화와 자외선과 같은 외부 환경에 노출되어 발생하는 광노화가 합쳐진 결과로 발생한다. 노화된 피부에서는 상처치유 능력과 면역기능, 비타민 D3 합성 능력, 땀과 피지 분비 능력이 감퇴되고 피부의 양성 및 악성종양의 발생이 증가하게 되는 등 전반적인 피부 기능의 변화가 나타난다.Skin aging occurs as a result of the combination of genetically determined endogenous aging and photoaging that occurs when exposed to external environments such as ultraviolet light. In aging skin, overall skin function changes such as wound healing ability, immune function, vitamin D3 synthesis ability, sweat and sebaceous secretion ability decline, skin positive and malignant tumors are increased.
특히 건조피부는 65세 이상의 노인층에서 최소한 75%에서 발생한다고 알려져 있을 정도로 노인에게 가장 흔한 피부과적 문제이다. 노인에서 건조피부의 발생 원인은 아직 명확하게 밝혀지지 않은 상태이나, 피부장벽기능의 장애와의 연관성에 대한 연구가 이루어지고 있다.Dry skin, especially dry skin, is known to occur in at least 75% of elderly people over 65 years old. The causes of dry skin in the elderly have not yet been clarified, but studies have been conducted on the relationship between skin barrier function and disability.
피부장벽기능 회복은 연령에 따라 차이가 있는데, 급성 장벽손상 시 젊은 사람의 경우 최초 12시간에 약 50%의 회복이 이루어지며 완전한 회복이 이루어지는 데는 약 3일 정도의 시간이 이 걸리는 반면, 노인의 경우는 완전한 회복이 이루어지는 데 약 1주일 정도가 소요된다. 이렇게 노화된 피부에서는 장벽기능의 회복 속도가 젊은 사람에 비해 저하되어 있음으로 해서 건조피부를 비롯하여 약물의 경피흡수의 변화, 자극성 접촉피부염에 대한 감수성 감소 등 표피의 여러 중요한 기능의 변화가 나타나게 된다.The recovery of skin barrier function varies with age. In the case of acute barrier damage, about 50% of recovery is achieved in the first 12 hours and about 3 days is needed for complete recovery. It takes about a week for a complete recovery to take place. In this aged skin, the recovery rate of the barrier function is lower than that of the young person, and various important function changes of the epidermis such as changes in the transdermal absorption of drugs, reduction of susceptibility to irritant contact dermatitis are observed.
노화피부에서 경표피 수분 손실(transepidermal water loss, TEWL)로 측정한 기초 상태의 피부 장벽 기능은 정상처럼 보이지만, 급성 피부장벽의 손상을 유발시킨 경우, 장벽이 쉽게 파괴되고, 회복이 지연된다. 또한 각질층간의 결합성이 떨어지고, 자외선에 대한 표피의 염증 반응도 감소된다.Skin barrier function of basal state measured by transepidermal water loss (TEWL) in aging skin seems normal, but if it causes acute skin barrier damage, barrier is easily broken and recovery is delayed. Also, the bond between the stratum corneum is lowered, and the inflammation reaction of the epidermis against ultraviolet light is also decreased.
피부 각질세포내의 필라그린이란 단백질이 분해되면 여러 자연보습인자가 형성되어 각질층의 수분을 유지해 주는데, 노인의 경우 필라그린 단백질이 분해가 저하되어 각질층의 수분 함량이 저하되며 피부 장벽을 이루고 있는 지질도 급격히 감소되어 피부 건조증이 발생하게 된다.In the case of elderly people, the degradation of fila green protein is reduced and the water content of the stratum corneum is lowered. Also, the lipid constituting the skin barrier is rapidly reduced Resulting in dry skin.
노화피부에서는 각질층의 수분 함유량이 유의하게 감소하여 있는데, 각질층지질구성 성분의 감소, 자연보습인자의 감소, glycerol 양의 감소 등이 그 원인으로 제시되고 있다. 각질층 내의 glycerol의 감소는 노화에 따른 피지 분비의 감소와 표피세포의 수분/glycerol 수송체(aquaporin-3)의 감소로 인해 발생한다고 알려져 있다.In the aging skin, the water content of the stratum corneum is decreased significantly. The decrease of the stratum corneum lipid component, the decrease of the natural moisturizing factor and the decrease of the glycerol amount are suggested as the causes. It is known that the decrease of glycerol in the stratum corneum is caused by the decrease of sebum secretion and the decrease of water / glycerol transporter (aquaporin-3) in the epidermal cells with aging.
결국 노화피부에서는 하부 표피로부터 각질층까지 공급되는 수분량이 부족하여 피부 건조가 심해지고 이로 인해 염증과 소양증이 쉽게 발생하게 되는 것이다.As a result, in the aging skin, the amount of water supplied from the lower epidermis to the stratum corneum is insufficient, so that the skin becomes dry and the inflammation and pruritus easily occur.
이렇게 노인의 피부 건조상태는 일차적으로 소양감과 불편감을 느끼게 하며 이차적으로는 감염과 욕창 등의 피부문제를 일으킨다. 이에 따른 불안, 초조, 수면장애, 자아개념장애 및 자기 역할장애 같은 심각한 정서장애까지 동반할 수 있어 노인의 피부 건강상태에 대한 관찰 및 관리가 중요하므로, 노인 피부 장벽의 회복을 위한 많은 연구와 치료제의 개발이 요구되는 실정이다.The dryness of the skin of the elderly makes the first feelings of pruritus and discomfort, and secondly it causes skin problems such as infection and pressure ulcer. Therefore, it is important to observe and manage the skin health status of the elderly, as it can accompany serious emotional disorders such as anxiety, irritability, sleeping disorder, self-conceptual disorder, and self-role disorder. Therefore, Is required.
종래 기술의 문제점을 해결하기 위해, 본 발명은 피토스핑고신을 유효성분으로 함유하는, 노인성 건조증 예방 또는 치료용 약학적 조성물 또는 개선용 건강식품 조성물을 제공하는 것이다.In order to solve the problems of the prior art, the present invention is to provide a pharmaceutical composition or health food composition for prevention or treatment of dryness of senile dryness containing phytosphingosine as an active ingredient.
1. 피토스핑고신을 유효성분으로 함유하는, 노인성 건조증 예방 또는 치료용 약학적 조성물.1. A pharmaceutical composition for preventing or treating senile dryness comprising phytosphingosine as an active ingredient.
2. 위 1에 있어서, 상기 피토스핑고신은 프로필라그린 및 필라그린 중 적어도 하나의 발현을 촉진하는 것인, 노인성 건조증 예방 또는 치료용 약학적 조성물.2. The pharmaceutical composition according to 1 above, wherein said phytosphingosine promotes the expression of at least one of propylaguanine and pilagreen.
3. 위 1에 있어서, 상기 피토스핑고신은 카스파제-14(Caspase-14)와 블레오마이신 하이드로라제 발현을 유도하여 필라그린의 분해를 촉진하는 것인, 약학적 조성물.3. The pharmaceutical composition according to 1 above, wherein said phytosphingosine induces caspase-14 (caspase-14) and bleomycin hydrolyzate expression to promote decomposition of filaggrin.
4. 위 1에 있어서, 상기 피토스핑고신은 세라마이드 합성효소-3(CerS-3)의 발현을 촉진하는 것인, 노인성 건조증 예방 또는 치료용 약학적 조성물.4. The pharmaceutical composition according to 1 above, wherein said phytosphingosine promotes expression of ceramide synthase-3 (CerS-3).
5. 위 1에 있어서, 상기 피토스핑고신은 클라우딘의 발현을 촉진하는 것인, 노인성 건조증 예방 또는 치료용 약학적 조성물.5. The pharmaceutical composition for preventing or treating dry skin according to 1 above, wherein said phytosphingosine promotes expression of claudin.
6. 위 1에 있어서, 상기 노인성 건조증은 아토피 및 어린선 중 적어도 하나인, 노인성 건조증 예방 또는 치료용 약학적 조성물.6. The pharmaceutical composition for preventing or treating dry skin according to 1 above, wherein the senile dry symptom is at least one of atopy and lice.
7. 피토스핑고신을 유효성분으로 함유하는, 노인성 건조증 예방 또는 개선용 건강식품 조성물.7. A health food composition for prevention or improvement of senile dryness comprising phytosphingosine as an active ingredient.
8. 위 7에 있어서, 상기 피토스핑고신은 프로필라그린 및 필라그린 중 적어도 하나의 발현을 촉진하는 것인, 노인성 건조증 예방 또는 개선용 건강식품 조성물.8. The health food composition according to 7 above, wherein the phytosphingosine promotes the expression of at least one of propylaguanine and pilagreen.
9. 위 7에 있어서, 상기 피토스핑고신은 카스파제-14(Caspase-14)와 블레오마이신 하이드로라제 발현을 유도하여 필라그린의 분해를 촉진하는 것인, 건강식품 조성물.9. The health food composition according to 7 above, wherein said phytosphingosine induces caspase-14 (caspase-14) and bleomycin hydrolase expression to promote degradation of filaggrin.
10. 위 7에 있어서, 상기 피토스핑고신은 세라마이드 합성효소-3(CerS-3)의 발현을 촉진하는 것인, 노인성 건조증 예방 또는 개선용 건강식품 조성물.10. The health food composition according to 7 above, wherein said phytosphingosine promotes expression of ceramide synthase-3 (CerS-3).
11. 위 7에 있어서, 상기 피토스핑고신은 클라우딘의 발현을 촉진하는 것인, 노인성 건조증 예방 또는 개선용 건강식품 조성물.11. The health food composition for prevention or improvement of senile dryness according to 7 above, wherein said phytosphingosine promotes expression of claudin.
12. 위 7에 있어서, 상기 노인성 건조증은 아토피 및 어린선 중 적어도 하나인, 노인성 건조증 예방 또는 개선용 건강식품 조성물.12. The composition for preventing or treating senile dryness according to 7 above, wherein the senile dry symptom is at least one of atopic and astringent.
본 발명에 따른 피토스핑고신을 포함하는 조성물은 필라그린 단백질이 분해가 저하되거나, 세라마이드 합성효소-3(CerS-3)의 발현이 저하되어 나타나는 노인성 건조증을 개선시킬 수 있는 효과가 있으므로, 노인성 건조증을 치료 또는 개선하는데 유용하게 사용될 수 있다.The composition comprising phytosphingosine according to the present invention has an effect of improving the dryness of senile dryness which is caused by degradation of filamentous green protein or degradation of expression of ceramide synthase-3 (CerS-3) Can be usefully used to treat or ameliorate dryness.
도 1은 본 발명에 따른 피토스핑고신의 필라그린 단백질 발현에 미치는 효과 분석한 결과를 나타내는 도면이다.
도 2는 본 발명에 따른 피토스핑고신의 카스파제-14와 블레오마이신 하이드로라제 단백질 발현에 미치는 영향을 분석한 결과를 나타내는 도면이다.
도 3은 본 발명에 따른 피토스핑고신의 필라그린 및 카스파제-14의 mRNA 발현에 미치는 영향을 분석한 결과를 나타내는 도면이다.
도 4는 본 발명에 따른 피토스핑고신의 세라마이드 합성효소 3(CerS 3)의 발현에 미치는 영향을 분석한 결과를 나타내는 도면이다.
도 5는 본 발명에 따른 피토스핑고신의 Tight Junction 관련 단백질의 발현에 미치는 영향을 분석한 결과를 나타내는 도면이다.
도 6은 본 발명에 따른 피토스핑고신을 포함한 조성물의 노인성 건조증 개선 효과를 나타내는 도면이다.FIG. 1 is a graph showing the results of an analysis of the effect of Phytosphingosine on the expression of filaggrin protein according to the present invention.
FIG. 2 is a graph showing the results of analysis of the effect of phytosphingosine on expression of caspase-14 and bleomycin hydrolyzate protein according to the present invention. FIG.
FIG. 3 is a graph showing the results of analyzing the effect of Phytosphingosine on the expression of mRNA of filaggrin and caspase-14 according to the present invention.
FIG. 4 is a graph showing the results of analysis of the effect of the ceramide synthase 3 (CerS 3) on the expression of phytosphingosine according to the present invention.
FIG. 5 is a graph showing the results of analyzing the effect of the expression of the tight junction-related protein of Phytosphingosine according to the present invention.
FIG. 6 is a graph showing the effect of improving the dry feeling of senescence in the composition containing phytosphingosine according to the present invention. FIG.
이하에서, 본 발명의 여러 측면 및 다양한 구현예에 대해 더욱 구체적으로 살펴보도록 한다. Hereinafter, various aspects and various embodiments of the present invention will be described in more detail.
본 발명은, 피토스핑고신을 유효성분으로 함유하는, 노인성 건조증 예방 또는 치료용 약학적 조성물을 제공한다.The present invention provides a pharmaceutical composition for preventing or treating dry skin of the senescence comprising phytosphingosine as an active ingredient.
종래의 피부 건조증을 개선하는 소재는 세라마이드, 콜레스테롤, 유리지방산을 1:1:1 또는 2:1:1의 비율로 혼합한 생리적 피부 지질 혼합물을 주로 사용하였으며, 이를 통해 피부의 보습이 증가하고 손상된 피부장벽의 기능이 빠르게 회복되는 효과를 가져왔다. 그러나 이러한 피부지질 혼합물은 근본적인 피부의 항상성을 정상적으로 되돌리는 데는 한계가 있으며 외부에서 도포함으로 부족한 피부장벽간 지질을 보충해주는 효과를 가지고 있어 지속적인 효과를 보기는 어려움이 있다.Conventional skin dryness enhancement materials are mainly composed of physiological skin lipid mixture of ceramide, cholesterol, and free fatty acid in a ratio of 1: 1: 1 or 2: 1: 1, The function of the skin barrier has been rapidly restored. However, such a skin lipid mixture has a limitation in restoring the intrinsic homeostasis of the skin normally, and it is difficult to see a continuous effect because it has an effect of supplementing the lipid between skin barrier which is insufficient by applying it from the outside.
특히 노화피부에서 경표피 수분 손실(transepidermal water loss, TEWL)로 측정한 기초 상태의 피부 장벽 기능은 정상처럼 보이지만, 급성 피부장벽의 손상을 유발시킨 경우, 장벽이 쉽게 파괴되고, 회복이 지연된다. 또한 각질층간의 결합성이 떨어지고, 자외선에 대한 표피의 염증 반응도 감소되는 만큼, 노인의 경우 피부가 손상된 후 피부의 장벽이 회복되는 속도가 떨어지므로 젊은 사람들에 비해 피부 건조증이 오래 발생한다.In particular, the basal skin barrier function measured by transepidermal water loss (TEWL) in aging skin seems normal, but if it causes acute skin barrier damage, the barrier is easily broken and recovery is delayed. In addition, as the bond between the stratum corneum is lowered and the inflammation reaction of the epidermis against the ultraviolet rays is decreased, the dry skin of the elderly is longer than the younger ones because the skin barrier is restored after the skin is damaged.
본 발명에 따른 피토스핑고신은 하기 화학식 1로 표시되는 화합물로서, 최초로 식물에서 추출되었으며 그 구조가 스핑고신과 유사하였기 때문에 피토스핑고신이라는 이름이 붙여졌다. 최근 곰팡이, 효모 등의 미생물뿐 만이 아니라 해양 생물, 특히 포유 동물의 조직에서 많이 발견되고 있다.The phytosphingosine according to the present invention is a compound represented by the following formula (1), which was first extracted from a plant and named Phytosphingosine because its structure was similar to sphingosine. Recently, microorganisms such as molds and yeasts are found not only in marine organisms, but also in mammalian tissues.
[화학식 1][Chemical Formula 1]
최근에 각질층 형성과정에 관한 연구가 많이 발전하면서 각질세포형성에 필라그린이 핵심적인 기능을 수행하는 것으로 알려지고 있다. 최근의 연구에 의하면 필라그린(Filaggrin: FLG)은 필라멘트 응집 단백질(filament aggregating protein)로 표피의 과립층(stratum granulosom)에 주로 위치하며 케라토하이알린(keratohyaline granules)을 이루며 약 400 kDa 크기의 프로필라그린(Profilaggrin) 상태로 존재한다. 프로필라그린은 10~12개의 필라그린이 결합된 중합체로 각질세포 분화 시 탈인산되고 분해되어 필라그린 단량체(monomer)가 된다. 이후 필라그린은 카스파제-14(Caspase-14), 펩티딜알르기닌 디이미나제(peptidylarginine deiminase), 블레오마이신 하이드로라제(bleomycin hydrolase) 등의 단백질 분해효소에 의해 글루타민(glutamine), 아르기닌(arginine), 히스티딘(histidine) 등으로 분해되고, 이후 더욱 분해되어 acidic polycarboxylic acid osmolytes를 생성하여 각질층의 보습(천연보습인자: Natural moisturizing factor)을 담당한다.Recently, studies on the process of stratum corneum formation have been developed and it is known that pillar green plays a key role in keratinocyte formation. Recent studies have shown that filaggrin (FLG) is a filament aggregating protein that is mainly located in the stratum granulosomes of the epidermis and forms keratohyaline granules and has a profile of about 400 kDa It is present in the green (Profilaggrin) state. Propylguanine is a polymer with 10 to 12 pillagreenes attached. It is dephosphorylated upon keratinocyte differentiation and degraded to a filamentary monomer. Afterwards, pilagreen is converted to glutamine, arginine, and the like by proteolytic enzymes such as Caspase-14, peptidylarginine deiminase, and bleomycin hydrolase. , Histidine, etc., and then decomposed to produce acidic polycarboxylic acid osmolytes, which are responsible for the moisturizing factor (natural moisturizing factor) of the stratum corneum.
한편, 카스파제-14는 피부장벽세포 형성과정에서 프로필라그린(Profilaggrin)을 분해하는 효소이다. 프로필라그린의 분해에 문제가 발생하면 피부에서 천연보습인자의 생성이 감소하여 피부에 문제가 발생하고, 카스파제-14는 최종 각질세포 분화단계에서 핵심적인 역할을 수행하는데 카스파제-14에 문제가 발생하면 번들거리는 피부(shinier skin)가 되고 경표피수분증발이 증가하며, 피부의 보습에 문제가 발생한다.On the other hand, caspase-14 is an enzyme that degrades profilaggrin in the process of skin barrier cell formation. Problems with the degradation of propylaguanine cause a reduction in the production of natural moisturizing factors in the skin, causing skin problems, and caspase-14 plays a key role in the final stage of keratinocyte differentiation. , The skin becomes shinier skin, the transpulmonary water evaporation increases, and the problem of moisturization of the skin occurs.
노인의 경우에 이러한 필라그린 단백질의 분해가 저하되는 문제점이 있어 건조증을 야기하는데, 본 발명에 따르면, 피토스핑고신은 프로필라그린의 발현을 촉진하고, 카스파제-14와 블레오마이신 하이드로라제 발현을 유도하여 필라그린의 분해를 촉진하는 효과가 있어 노인성 건조증을 치료 또는 개선하는데 유용하게 사용될 수 있다.In the case of the elderly, degradation of the pilar green protein is degraded, resulting in dryness. According to the present invention, phytosphingosine promotes the expression of propyllaurine and induces expression of caspase-14 and bleomycin hydrolase And promotes degradation of pilgar green. Thus, it can be effectively used to treat or ameliorate senile dryness.
또한 피부 각질 세포간 지질의 주요 성분은 세라마이드와 콜레스테롤, 유리지방산등이 규칙적인 층상구조(Lamella structure)를 이루며 각질세포의 사이를 빈틈없이 채우고 있어야 한다.In addition, the main component of intercellular lipid bilayer is ceramide, cholesterol, free fatty acid, etc., which form a regular lamella structure and fill the space between keratinocytes tightly.
장벽세포간 지질 중 가장 많이 존재하는 성분이 세라마이드인데 피부에 존재하는 세라마이드는 장쇄의 스핑고지질 염기와 여기에 결합된 지방산의 종류에 따라 다양하게 분류되며, 피부에 존재하는 종류도 수백종에 이르는 것으로 알려져 있다. 세라마이드는 결합된 지방산의 길이에 따라 구분된다. 이중 C22 이상의 세라마이드의 합성에 관여하는 세라마이드 합성효소-3(CerS-3)에 문제가 발생하면 피부장벽에 문제가 발생한다.Barrier cell interstitial lipid is the most abundant component of ceramide. Ceramide present in skin is classified into various kinds according to kinds of long chain sphingolipid base and fatty acid bonded thereto, and there are hundreds of species present in skin It is known. Ceramides are distinguished by the length of the fatty acids bound. Among them, when problems occur in ceramide synthase-3 (CerS-3) involved in the synthesis of C22 or higher ceramide, problems occur in the skin barrier.
또한, 노인의 경우 세라마이드 합성효소-3(CerS-3)의 발현이 저하되는 문제점이 있어 노인 건조증을 야기하는데, 본 발명에 따르면, 피토스핑고신은 세라마이드 합성효소-3(CerS-3)의 발현을 촉진하고, 클라우딘의 발현을 촉진하는 효과가 있어 세라마이드 합성효소-3(CerS-3)의 발현이 저하되어 나타나는 노인성 건조증을 개선시킬 수 있는 효과가 있으므로, 노인성 건조증을 치료 또는 개선하는데 유용하게 사용될 수 있다.In addition, in the elderly, the expression of ceramide synthase-3 (CerS-3) is lowered and causes dryness of the elderly. According to the present invention, phytosphingosine is produced from ceramide synthase- (CerS-3), which is effective for improving senescence of senile dryness, which is effective for the treatment or improvement of senile dry symptom. Can be used.
따라서 본 발명에 따른 피토스핑고신은 약학적으로 허용가능한 염을 포함하여 노인성 건조증 예방 또는 치료용 약학적 조성물로 적용될 수 있고, 이외에도 피토스핑고신을 유효성분으로 함유하는 노인성 건조증 예방 또는 개선용 건강식품 조성물이나 화장료 조성물에 적용될 수 있다.Accordingly, the phytosphingosine according to the present invention can be applied as a pharmaceutical composition for the prevention or treatment of senile dryness including pharmacologically acceptable salts. In addition, phytosphingosine can be used for prevention or improvement of senile dryness containing phytosphingosine as an active ingredient It can be applied to a health food composition or a cosmetic composition.
상기 본 발명에 따른 피토스핑고신을 포함하는 노인성 건조증 예방 또는 치료용 약학적 조성물은 방부제, 안정화제, 수화제 또는 유화 촉진제, 삼투압 조절을 위한 염 및/또는 완충제 등의 약제학적 보조제 및 기타 치료적으로 유용한 물질을 추가로 함유할 수 있으며, 통상적인 방법에 따라 다양한 경구 투여제 또는 비경구 투여제 형태로 제형화할 수 있다.The pharmaceutical composition for preventing or treating dry skin of the senile comprising phytosphingosine according to the present invention is useful as a preservative, a stabilizer, a wetting agent or an emulsifying accelerator, a pharmaceutical adjuvant such as a salt and / or a buffer for controlling osmotic pressure, , And may be formulated into various oral or parenteral dosage forms according to conventional methods.
상기 경구 투여제는 예를 들면, 정제, 환제, 경질 및 연질 캅셀제, 액제, 현탁제, 유화제, 시럽제, 분제, 산제, 세립제, 과립제, 펠렛제 등이 있으며, 이들 제형은 유효성분 이외에 계면 활성제, 희석제(예: 락토즈, 덱스트로즈, 수크로즈, 만니톨, 솔비톨, 셀룰로즈 및 글리신), 활택제(예: 실리카, 탈크, 스테아르산 및 그의 마그네슘 또는 칼슘염 및 폴리에틸렌 글리콜)를 함유할 수 있다. 정제는 또한 마그네슘 알루미늄 실리케이트, 전분 페이스트, 젤라틴, 트라가칸스, 메틸셀룰로즈, 나트륨 카복시메틸셀룰로즈 및 폴리비닐피롤리딘과 같은 결합제를 함유할 수 있으며, 경우에 따라 전분, 한천, 알긴산 또는 그의 나트륨 염과 같은 붕해제, 흡수제, 착색제, 향미제, 및 감미제 등의 약제학적 첨가제를 함유할 수 있다. 상기 정제는 통상적인 혼합, 과립화 또는 코팅 방법에 의해 제조될 수 있다.Examples of the oral administration agent include tablets, pills, hard and soft capsules, liquids, suspensions, emulsions, syrups, powders, powders, granules, granules and pellets, , Diluents (such as lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and glycine), lubricants (such as silica, talc, stearic acid and magnesium or calcium salts thereof and polyethylene glycols) . Tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and polyvinylpyrrolidine, optionally mixed with starch, agar, alginic acid or its sodium salt Such as disintegrants, absorbents, coloring agents, flavoring agents, and sweetening agents. The tablets may be prepared by conventional mixing, granulating or coating methods.
또한, 상기 비경구 투여제는 예를 들어, 주사제, 점적제, 연고, 로션, 겔, 크림, 스프레이, 현탁제, 유제, 좌제(坐劑), 패취 등의 제형일 수 있으나, 이에 제한되는 것은 아니다.The parenteral administration may be, for example, an injection, a drip, an ointment, a lotion, a gel, a cream, a spray, a suspension, an emulsion, a suppository, no.
본 발명에 따른 상기 약학 조성물은 경구, 비경구, 직장, 국소, 경피, 정맥 내, 근육 내, 복강 내, 피하 등으로 투여될 수 있다.The pharmaceutical composition according to the present invention may be administered orally, parenterally, rectally, topically, transdermally, intravenously, intramuscularly, intraperitoneally, subcutaneously and the like.
또한, 상기 활성성분의 투여량은 치료 받을 대상의 연령, 성별, 체중과, 치료할 특정 질환 또는 병리 상태, 질환 또는 병리 상태의 심각도, 투여경로 및 처방자의 판단에 따라 달라질 것이다. 이러한 인자에 기초한 투여량 결정은 당업자의 수준 내에 있다. 일반적인 투여량은 0.001mg/kg/일 내지 2000mg/kg/일, 보다 구체적으로는 0.5mg/kg/일 내지 2.5mg/kg/일이다.In addition, the dosage of the active ingredient will depend on the age, sex, body weight of the subject to be treated, the particular disease or condition to be treated, the severity of the disease or condition, the route of administration and the judgment of the prescriber. Dosage determinations based on these factors are within the level of those skilled in the art. Typical dosages are from 0.001 mg / kg / day to 2000 mg / kg / day, more specifically from 0.5 mg / kg / day to 2.5 mg / kg / day.
또한 본 발명에 따른 피토스핑고신을 포함하는 노인성 건조증 예방 또는 개선용 건강식품 조성물은 상기 건강식품 조성물은 제형이 특별히 한정되지 않으나, 예를 들어, 정제, 과립제, 드링크제, 캬라멜, 다이어트바 등으로 제형화 될 수 있다. 각 제형의 건강식품 조성물은 유효 성분 이외에 해당 분야에서 통상적으로 사용되는 성분들을 제형 또는 사용 목적에 따라 당업자가 어려움 없이 적의 선정하여 배합할 수 있으며, 다른 원료와 동시에 적용할 경우 상승 효과가 일어날 수 있다.In addition, the health food composition for prevention or improvement of senile dryness comprising phytosphingosine according to the present invention is not particularly limited as far as formulation of the health food composition is applied. For example, tablet, granule, drink, caramel, Can be formulated. The health food composition of each formulation may be formulated by those skilled in the art without difficulty, depending on the purpose of formulation or use, in addition to the active ingredient, and the synergistic effect may occur when the composition is applied simultaneously with other ingredients .
이하에서 실시예 등을 통해 본 발명을 더욱 상세히 설명하고자 하며, 다만 이하에 실시예 등에 의해 본 발명의 범위와 내용이 축소되거나 제한되어 해석될 수 없다. 또한, 이하의 실시예를 포함한 본 발명의 개시 내용에 기초한다면, 구체적으로 실험 결과가 제시되지 않은 본 발명을 통상의 기술자가 용이하게 실시할 수 있음은 명백한 것이며, 이러한 변형 및 수정이 첨부된 특허청구범위에 속하는 것도 당연하다.Hereinafter, the present invention will be described in more detail with reference to Examples and the like, but the scope and content of the present invention can not be construed to be limited or limited by the following Examples. It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit and scope of the present invention as set forth in the following claims. It is natural that it belongs to the claims.
실시예1Example 1 : : 피토스핑고신의Phytosphingosine 필라그린Pillar Green 단백질 발현에 미치는 효과 분석 Analysis of effect on protein expression
Human epidermal keratinocytes (NEKn)을 EpiLife medium을 이용해 배양한 후 6 well plate에 2x105으로 파종하여 24시간 배양 한 후, 피토스핑고신을 각각 1uM, 5uM의 농도로 처리하고 72 시간 동안 추가 배양하였다. 배양이 끝난 후 케라틴세포(Keratinocyte)를 회수하고 파쇄한 후 동일양의 단백질을 SDS-PAGE에 전기영동 한 후 FLG 항체를 이용해 웨스턴 블랏 분석을 수행하였다.Human epidermal keratinocytes (NEKn) were cultured in EpiLife medium and seeded at 2 × 10 5 in a 6-well plate for 24 hours. Phytosphingosine was further treated at a concentration of 1 μM and 5 μM, respectively, for 72 hours. After incubation, keratinocytes were recovered and disrupted, and the same amount of protein was electrophoresed on SDS-PAGE, followed by Western blot analysis using FLG antibody.
그 결과 도 1에 나타낸 바와 같이, 피토스핑고신의 처리는 무처리군에 비해 프로필라그린의 발현과 필라그린의 분해를 증가시키는 것으로 확인되었다.As a result, as shown in Fig. 1, the treatment of phytosphingosine was confirmed to increase the expression of propylaguanine and the degradation of filaggrin in comparison with the untreated group.
프로필라그린의 발현과 필라그린 분해는 칼슘이온에 의해 각질세포가 분화되면서 증가하는 양과 비슷한 수준으로 증가하는 것을 확인되었고, 유사한 스핑고지질 골격인 Sphinganine(스핑가닌)과 Sphingosine(스핑고신)에 의해서도 필라그린의 발현이 증가하는 것을 확인하였다.It was found that the expression of propylaguanine and the degradation of filaggrin increased to a level similar to that of keratinocyte differentiation by calcium ion and that Sphinganine (sphinganine) and Sphingosine (sphingosine) The expression of pillar green was also increased.
실시예Example 2: 2: 피토스핑고신의Phytosphingosine 카스파제Caspase -14와 -14 and 블레오마이신Bleomycin 하이드로라제Hydro lase 단백질 발현에 미치는 영향 분석 Analysis of effect on protein expression
Human epidermal keratinocytes (NEKn)을 EpiLife medium을 이용해 배양한 후 6 well plate에 2x105으로 파종하여 24시간 배양 한 후 피토스핑고신을 각각 1uM, 5uM의 농도로 처리하고 72 시간 동안 추가 배양하였다. 배양이 끝난 후 케라틴세포를 회수하고 파쇄한 후 동일양의 단백질을 SDS-PAGE에 전기영동 한 후 카스파제-14와 블레오마이신 하이드로라제 (BLMH) 항체를 이용해 웨스턴 블랏 분석을 수행하였다.Human epidermal keratinocytes (NEKn) were cultured in EpiLife medium and seeded at 2 × 10 5 in a 6-well plate for 24 hours. Phytosphingosine was further treated at a concentration of 1 μM and 5 μM, respectively, for 72 hours. After cultivation, keratinocytes were recovered and disrupted, and the same amount of protein was electrophoresed on SDS-PAGE, followed by Western blot analysis using caspase-14 and a bleomycin hydrolyzate (BLMH) antibody.
그 결과 도 2a 및 도 2b에 나타낸 바와 같이, 피토스핑고신의 처리는 무처치군에 비해 필라그린의 분해에 관여하는 카스파제 14과 블레오마이신 하이드로라제의 발현을 증가시키는 것을 확인하였다.As a result, as shown in Figs. 2A and 2B, it was confirmed that the treatment of phytosphingosine increased expression of
전술한 바와 같이, 실시예 1과 실시예 2를 종합해보면 피토스핑고신의 처리는 각질 세포내 필라그린의 발현을 증가시키고, 동시에 Caspase-14, 블레오마이신 하이드로라제 의 발현을 증가시킴으로 필라그린의 분해를 원활히 하여 각질층의 온전한 형성과 각질층 세포에서의 천연보습인자의 양을 증가시키는 효과를 나타냄을 알 수 있다.As described above, the combination of Example 1 and Example 2 shows that the treatment of phytosphingosine increases the expression of pillar green in keratinocytes and at the same time increases expression of caspase-14 and pemomycin hydrolase, It can be seen that the decomposition is smoothly performed, and the effect of increasing the amount of the natural moisturizing factor in the stratum corneum cell and the intact formation of the stratum corneum is shown.
실시예Example 3: 3: 피토스핑고신의Phytosphingosine 필라그린Pillar Green 및 And 카스파제Caspase -14의 -14 mRNAmRNA 발현에 미치는 영향 분석 Analysis of effects on expression
Human epidermal keratinocytes (NEKn)을 EpiLife medium을 이용해 배양한 후 60 파이 dish 에 4x105으로 파종하여 24시간 배양 한 후 피토스핑고신을 처리해 48시간 동안 추가 배양하였다. 배양이 끝난 후 mRNA 분리 후 cDNA를 합성하였다. 합성한 cDNA와 각각의 프라이머를 이용하여 quantitative real-time PCR을 수행하였다.Human epidermal keratinocytes (NEKn) were cultured in EpiLife medium, seeded at 4 × 10 5 in a 60-pie dish, cultured for 24 hours, and further cultured for 48 hours with phytosphingosine treatment. After cultivation, mRNA was isolated and cDNA was synthesized. Quantitative real-time PCR was performed using the synthesized cDNA and each primer.
그 결과 도 3에 나타낸 바와 같이, 실시예 1과 실시예 2의 결과를 전사 레벨(mRNA transcription level)에서 확인한 결과 동일한 결과를 얻었다. 즉 필라그린과 카스파제-14는 피토스핑고신의 처리에 의해 농도의존적으로 발현량이 증가하는 것을 알 수 있다.As a result, as shown in Fig. 3, the results of Example 1 and Example 2 were confirmed at the transcription level (mRNA transcription level), and the same results were obtained. That is, the expression level of pilase green and caspase-14 increased in a concentration-dependent manner by treatment with phytosphingosine.
실시예Example 4: 4: 피토스핑고신의Phytosphingosine 세라마이드 합성효소 3( Ceramide synthetic enzyme 3 ( CerSCerS 3)의3) of 발현에 미치는 영향분석 Analysis of effects on expression
Human epidermal keratinocytes (NEKn)을 EpiLife medium을 이용해 배양한 후 6well plate에 2x105으로 파종하여 24시간 배양 한 후 피토스핑고신을 처리해 72 시간 동안 추가 배양하였다. 배양이 끝난 후 cell lysate을 회수하고 동일양의 단백질을 SDS-PAGE 한 후 CERS3 antibody를 이용해 웨스턴 블랏 분석을 수행하였다.Human epidermal keratinocytes (NEKn) were cultured in EpiLife medium, seeded at 2 × 10 5 on a 6- well plate, cultured for 24 hours, and further cultured for 72 hours with phytosphingosine treatment. After incubation, cell lysate was recovered and the same amount of protein was subjected to SDS-PAGE and Western blot analysis was performed using CERS3 antibody.
그 결과, 도 4에 나타낸 바와 같이, 피토스핑고신은 세라마이드 합성효소 3(CerS 3)의 발현을 증가시키는 것을 확인하였다. 이는 피부 세포간 지질에서 장쇄의 세라마이드(C22 이상의 장쇄의 지방산이 결합된 세라마이드)의 함량을 증가시켜 피부장벽기능을 증가시키는 효과를 나타낼 것으로 판단된다.As a result, as shown in Fig. 4, it was confirmed that phytosphingosine increased the expression of ceramide synthase 3 (CerS 3). It is considered that this would increase the skin barrier function by increasing the content of long chain ceramide (ceramide having long chain fatty acid bound to C22 or more) in interstitial lipid of skin.
실시예Example 5: 5: 피토스핑고신의Phytosphingosine Tight Junction 관련 단백질의 발현에 미치는 영향 분석 Tight junction-related protein expression
Human epidermal keratinocytes (NEKn)을 EpiLife medium을 이용해 배양한 후 6 well plate에 2x105으로 파종하여 24시간 배양 한 후 피토스핑고신을 처리해 48시간 동안 추가 배양하였다. 배양이 끝난 후 cell lysate을 회수하고 동일양의 단백질을 이용해 SDS-PAGE 후 Claudin-1과 occludin antibody를 이용해 웨스턴 블랏 분석을 수행하였다.Human epidermal keratinocytes (NEKn) were cultured in EpiLife medium, seeded at 2 × 10 5 on a 6-well plate, cultured for 24 hours, and further cultured for 48 hours with phytosphingosine treatment. Cell lysate was recovered and SDS-PAGE was performed using the same amount of protein, followed by Western blot analysis using Claudin-1 and occludin antibody.
그 결과 도 5에 나타낸 바와 같이, 피토스핑고신은 Tight Junction에서 중요한 기능을 수행하는 단백질인 오클루딘과 클라우딘의 발현을 증가시키는 것을 확인하였다. 이는 2차 피부장벽이라 불리는 Tight junction의 결합을 더욱 견고히 하여 세포간 공간에서의 수분이나 유해물질의 이동을 억제하는 효과를 나타낸다.As a result, as shown in Fig. 5, it was confirmed that phytosphingosine increased the expression of the proteins euclidin and cladin, which perform important functions in tight junctions. This further strengthens the binding of the tight junction called the secondary skin barrier, thereby suppressing the movement of moisture and harmful substances in the intercellular space.
실시예Example 6: 노인성 건조증 개선 효과 6: Improvement of senile dryness
건조한 피부를 갖는 50대 이상의 남녀 5명을 대상으로 피토스핑고신 1% 수용액을 상박에 하루 1회 2주 동안 도포 하였다.A total of 5 men and women aged 50 or more with dry skin were applied a 1% aqueous solution of phytosphingosine in a suppository for 2 weeks once a day.
2주 후, 피험자의 경표피 수분 손실량(TEWL), 수분함량 및 PCA(pyrrolidone carboxylic acid) 변화량을 측정하였고, 그 결과를 도 6에 나타내었다. Non은 무처리 대조군, Vehicle은 0.5% 락틱 수용액, PHS는 0.5% 락틱 수용액에 피토스핑고신(PHS)를 1%로 녹인 수용액을 처리한 실험군을 말한다. After two weeks, the amount of water-loss (TEWL), water content and pyrrolidone carboxylic acid (PCA) of the subjects was measured. The results are shown in Fig. Non-treated control group, Vehicle 0.5% lactic aqueous solution, PHS 0.5% lactic aqueous solution was treated with an aqueous solution of 1% PHS dissolved in 1% solution.
도 6(A)는 경표피 수분 손실량(TEWL)를 측정한 결과를 나타낸 것이다. PHS는 무처리 대조군(Non) 보다 경표피 수분 손실량(TEWL)이 상대적으로 낮아졌음을 관찰할 수 있었다. 즉, 피토스핑고신은 피부의 수분 증발을 지연시킴으로 보습효과를 나타냄을 알 수 있다. Fig. 6 (A) shows the result of measuring the amount (TEWL) of the transdermal skin moisture loss. PHS showed relatively lower TEWL than non-treated control (Non). In other words, it can be seen that the phytosphingosine shows a moisturizing effect by delaying the water evaporation of the skin.
도 6(B)는 피토스핑고신에 의한 수분함량을 코네오미터(corneometer)로 측정하여 나타낸 결과이다. 피토스핑고신 수용액을 도포한 군(PHS)은 무처리 대조군(Non)과 비교했을 때 수분함량이 더 큰 것을 확인하였다. FIG. 6 (B) is a result of measuring the water content by phytosphingosine with a corneometer. (PHS) coated with an aqueous solution of phytosphingosine was found to have a higher water content than the non-treated control (Non).
도 6(C)는 피토스핑고신 수용액을 도포한 후 각질층을 회수하여 피부 각질층에 함유된 천연 보습인자 중 PCA (pyrrolidone carboxylic acid) 변화량을 나타낸 결과이다. 피토스핑고신 수용액(PHS) 처리에 의해 PCA의 함량이 증가한 것을 확인하였다. 이로부터 피토스핑고신 수용액에 의해 피부 보습력이 증가되었음을 알 수 있다. FIG. 6 (C) is a graph showing the amount of PCA (pyrrolidone carboxylic acid) change in the natural moisturizing factor contained in the stratum corneum of the skin after the application of the aqueous solution of phytosphingosine and recovery of the stratum corneum. It was confirmed that the content of PCA was increased by treatment with phytosphingosine aqueous solution (PHS). From this, it can be seen that the skin moisturizing power is increased by the aqueous solution of phytosphingosine.
Claims (2)
A composition for enhancing natural moisturizing factors in isolated keratinocytes, comprising phytosphingosine.
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