KR20210123853A - Composition for treatment, prevention or reducing atopic dermatitis, psoriasis or dermatitis comprising ganoderic acid - Google Patents
Composition for treatment, prevention or reducing atopic dermatitis, psoriasis or dermatitis comprising ganoderic acid Download PDFInfo
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- KR20210123853A KR20210123853A KR1020200041577A KR20200041577A KR20210123853A KR 20210123853 A KR20210123853 A KR 20210123853A KR 1020200041577 A KR1020200041577 A KR 1020200041577A KR 20200041577 A KR20200041577 A KR 20200041577A KR 20210123853 A KR20210123853 A KR 20210123853A
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- KR
- South Korea
- Prior art keywords
- composition
- ganoderic acid
- acid
- present
- ganoderic
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- 239000000203 mixture Substances 0.000 title claims abstract description 89
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Abstract
Description
본 명세서는 신규한 가노데릭산의 제조 방법, 가노데릭산의 아토피, 건선 또는 피부염증의 치료 예방, 또는 경감용 조성물에 관한 것이다.The present specification relates to a novel method for producing ganoderic acid, and to a composition for preventing or alleviating atopy, psoriasis or skin inflammation of ganoderic acid.
만성 염증성 피부 질환은 환경적유전적인 유발 요인들에 의해 일어난 피부 과민 반응에 의해 나타나는 질환을 의미한다. 신체는 외부의 자극으로부터의 자가 보호를 위해 이물질을 제거하고 유발된 염증반응을 조절하는 면역 반응을 가지고 있는데, 면역계에 이상이 생길 경우 면역질환이 발생하게 된다. 염증성 피부 질환은 피부와 관련된 면역체계 반응에 의해 발생하는 것으로 알레르기 피부염, 알레르기성비염, 천식, 알레르기성 결막염, 아토피성 두드러기 등이 이에 포함된다 (Donald Y.M.Leung et al, J. Clin Invest., 113(5)651-657(2004)). Chronic inflammatory skin disease refers to a disease caused by skin hypersensitivity reactions caused by environmental and genetic triggers. The body has an immune response that removes foreign substances for self-protection from external stimuli and regulates the induced inflammatory response. Inflammatory skin diseases are caused by skin-related immune system reactions, and include allergic dermatitis, allergic rhinitis, asthma, allergic conjunctivitis, and atopic urticaria (Donald YMLeung et al, J. Clin Invest., 113). (5) 651-657 (2004)).
아토피 피부염의 경우 유전적인 영향이 큰 것으로 알려져 있으나, 2018년 건강보험심사평가원의 조사 결과에 따르면 아토피피부염으로 진료를 받은 국내 환자 수는 94만 2,927명에 달하며 이 중 20세 이상의 성인이 42.7%를 차지했다. 이러한 성인 아토피 피부염은 유전적인 영향보다는 주거 및 근무 환경의 변화, 사회적 스트레스 등에 의해 유발되는 경우에 의한 것으로 판단되었다. Although it is known that atopic dermatitis has a large genetic influence, according to a survey by the Health Insurance Review and Assessment Service in 2018, the number of domestic patients treated for atopic dermatitis reached 942,927, of which 42.7% were adults over 20 years old. occupied This adult atopic dermatitis was judged to be caused by changes in living and working environment, social stress, etc. rather than genetic influences.
다른 피부염 질환의 일종인 건선의 경우 피부 내 세포의 비정상적인 활동에 의한 질병으로 아직 발병 원인이 명확히 밝혀져 있지 않다. 건선(Psoriasis)은 은백색의 인설을 동반한 구진을 나타내는 흔한 자가면역 피부 질환으로 분포나 정도가 개인에 따라 전신성 박탈, 홍반, 소양감, 건조감 및 작열감 등 아주 다양한 증상을 나타낸다. 전세계적으로 발생하나 인종 및 종족간 발생 빈도에 현저한 차이를 보이며 임상 경과는 다양할 수 있지만 일반적으로 호전과 악화를 반복하면서 지속되는 형태를 보인다. 건선의 발병 기전은 아직 완전히 규명되지 않았지만 염증세포의 침윤, 면역 활성화 분자 증가, Th1 및 Th17 반응, 염증 관련 사이토카인의 비정상적인 변화 등 염증이 중요한 역할을 한다는 연구결과가 보고되고 있다. 이러한 건선은 각질을 만들어내는 피부 세포가 일반적인 정상 피부에 비해 지나치게 자극을 받아 빠르게 증가하는 것으로 이에 따라 비듬과 같은 각질이 겹겹히 발생한다. 아토피 피부염과 달리 발병률이 성인에게 높으며 주로 팔꿈치와 무릅에서 발생한다. 주요 증상은 좁쌀 같은 발진과 두꺼운 각질이 보고되어 있다. Psoriasis, another type of dermatitis, is a disease caused by abnormal activity of cells in the skin, and the cause of the disease is still unknown. Psoriasis is a common autoimmune skin disease that presents with papules accompanied by silvery-white scales. The distribution and severity of psoriasis varies from person to person, including systemic deprivation, erythema, pruritus, dryness, and burning sensation. Although it occurs worldwide, there is a marked difference in the frequency of occurrence between races and races, and although the clinical course may vary, it generally shows a continuous form with repeated improvement and deterioration. Although the pathogenesis of psoriasis has not yet been fully elucidated, research results suggesting that inflammation plays an important role, such as infiltration of inflammatory cells, increased immune activation molecules, Th1 and Th17 responses, and abnormal changes in inflammation-related cytokines, are being reported. In psoriasis, the skin cells that produce dead skin cells are overstimulated compared to normal skin and increase rapidly. Unlike atopic dermatitis, the incidence rate is high in adults and mainly occurs in the elbow and knee. The main symptoms are millet-like rash and thick keratin.
현재 사용되고 있는 피부 질환 치료제의 경우에는 알레르기 반응의 산물인 히스타민 분비를 억제하거나 알레르기 증상의 하나인 염증 반응을 억제하는 스테로이드제 및 항히스타민제를 처방하고 있다. 이러한 아토피 질환 치료 연고의 경우 근본적인 염증의 생성을 예방하고 억제하는 염증성 질환이 아니며, 장기간 사용시 부작용을 유발할 수 있다. 스테로이드제의 경우 비만, 당뇨, 고혈압, 우울증 등을 항히스타민제의 경우 우울증, 집중력 장애, 무기력증, 졸림, 성기능 장애 등의 부작용을 일으키는 것으로 알려져 있어 아토피 질환에 사용 가능한 천연물 소재의 발굴과 이를 통한 치료제 개발이 중요시되고 있다. In the case of a skin disease treatment currently used, steroids and antihistamines that suppress the secretion of histamine, a product of an allergic reaction, or an inflammatory reaction, which are one of the symptoms of allergy, are prescribed. In the case of such atopic disease treatment ointment, it is not an inflammatory disease that prevents and suppresses the generation of underlying inflammation, and may cause side effects when used for a long period of time. Steroids are known to cause side effects such as obesity, diabetes, high blood pressure, and depression, while antihistamines are known to cause side effects such as depression, concentration disorders, lethargy, sleepiness, and sexual dysfunction. this is being considered important.
피부 염증은 내부외부 요인에 의한 자극으로 피부 장벽에 이상이 발생함에 따라 유도되는 신호 분자인 사이토카인(cytokine)에 의해 시작된다. 사이토카인은 면역 세포인 T 세포 (T cell)와 대식세포 (macrophage)에서 주로 생성되며, 인터루킨 (interlukins), 종양괴사인자-α (tumor necrosis factor-α, TNF-α), 인터페론 감마 (interferon gamma, IFNγ) 등이 이에 속한다 (SM Opal et al, CHEST, 117(4) 1162-1172, 2000).Skin inflammation is initiated by cytokines, which are signaling molecules that are induced when an abnormality occurs in the skin barrier due to stimulation by internal and external factors. Cytokines are mainly produced in immune cells, T cells and macrophages, interleukins, tumor necrosis factor-α (tumor necrosis factor-α, TNF-α), interferon gamma , IFNγ) and the like (SM Opal et al, CHEST, 117(4) 1162-1172, 2000).
케모카인 (chemokine)은 상처난 조직, 세균이나 바이러스에 감염된 조직 등에서 분비되는 저분자 단백질의 일종으로 백혈구의 유인 작용 및 활성화 작용을 하며 분자내 4개의 시스테인 잔기에 따라 분류되어 크게 CXCL, CCL, CX3CL 및 XCL로 분류된다. 또한, S100A7, S100A8 및 S100A9등의 S100 칼슘 결합 단백질들의 경우 염색체 1q21의 S100 유전자 클러스터 내에 위치하고 있으며 건선과 관련된 것으로 알려져있다. A chemokine is a type of small-molecular protein secreted from wounded tissue, bacteria or virus-infected tissue, etc., and has an attracting and activating action for leukocytes. classified as In addition, S100 calcium-binding proteins such as S100A7, S100A8 and S100A9 are located in the S100 gene cluster of chromosome 1q21 and are known to be associated with psoriasis.
이에 피부 염증 질환 예방 및 치료 효과를 확인하기 위해서 케모카인 (CXCL8, CCL5, CCL20)과 사이토카인 (IL-6, IL-23/p19), S100 칼슘 결합 단백질들 (S100A7, S100A8, S100A9) 등의 요소 발현 억제가 중요시 되어진다.Therefore, in order to confirm the prevention and treatment effect of skin inflammatory diseases, chemokines (CXCL8, CCL5, CCL20), cytokines (IL-6, IL-23/p19), S100 calcium binding proteins (S100A7, S100A8, S100A9), etc. Expression inhibition is important.
그러나 현재 아토피, 건선 또는 피부염증 질환에 효과가 있는 물질로는 스테로이드제 또는 항히스타민제가 사용되고 있으며, 이러한 치료 물질은 상기에서 언급한 바와 같이 다양한 부작용을 일으키는 것으로 알려져 있다. 또한, 이러한 치료 물질은 상기 질환의 증상인 히스타민의 생성단계를 억제하고 있으므로, 근본적인 염증의 생성을 예방 또는 억제하는 효과는 나타내고 있지 못하다.However, steroids or antihistamines are currently used as substances effective for atopic dermatitis, psoriasis, or skin inflammatory diseases, and these therapeutic substances are known to cause various side effects as mentioned above. In addition, since these therapeutic substances inhibit the generation of histamine, which is a symptom of the disease, the effect of preventing or inhibiting the generation of underlying inflammation is not shown.
따라서, 아토피, 건선 또는 피부염증의 치료를 위해서 보다 근본적인 염증메커니즘의 억제와 함께 부작용이 감소된 치료제 개발이 필요하다.Therefore, for the treatment of atopic dermatitis, psoriasis or skin inflammation, it is necessary to develop a therapeutic agent with reduced side effects and suppression of a more fundamental inflammatory mechanism.
상기와 같은 문제를 해결하기 위하여, 본 발명자들은 천연물에서 추출한 성분 중 가노데릭산이 아토피, 건선 및 피부 염증을 일으키는 케모카인 (CXCL8, CCL5, CCL20)과 사이토카인 (IL-6, IL-23/p19), S100 칼슘 결합 단백질들 (S100A7, S100A8, S100A9) 등의 요소 발현 억제가 우수함을 확인하여 본원발명에 이르게 되었다.In order to solve the above problems, the present inventors, among the components extracted from natural products, chemokines (CXCL8, CCL5, CCL20) and cytokines (IL-6, IL-23/p19) that ganoderic acid causes atopy, psoriasis and skin inflammation , S100 calcium binding proteins (S100A7, S100A8, S100A9), such as to confirm the excellent suppression of the expression of the elements led to the present invention.
이에 본 발명의 일측면은 가노데릭산을 유효성분으로 포함하는 아토피, 건선 또는 피부염증의 예방, 치료 또는 경감용 조성물을 제공하고자 한다.Accordingly, one aspect of the present invention is to provide a composition for the prevention, treatment or alleviation of atopic dermatitis, psoriasis or skin inflammation comprising ganoderic acid as an active ingredient.
또한, 본 발명의 일측면은 가노데릭산을 유효성분으로 포함하는 아토피, 건선 또는 피부염증의 예방, 치료 또는 경감용 조성물의 제조방법을 제공하고자 한다.In addition, one aspect of the present invention is to provide a method for preparing a composition for preventing, treating or alleviating atopic dermatitis, psoriasis, or skin inflammation comprising ganoderic acid as an active ingredient.
또한, 본 발명의 일측면은 아토피, 건선 또는 피부염증의 예방, 치료 또는 경감용 가노데릭산의 제조방법을 제공하고자 한다.In addition, one aspect of the present invention is to provide a method for preparing ganoderic acid for the prevention, treatment or alleviation of atopic dermatitis, psoriasis or skin inflammation.
본 발명의 목적은 이상에서 언급한 목적으로 제한되지 않는다. 본 발명의 목적은 이하의 설명으로 보다 분명해 질 것이며, 특허청구범위에 기재된 수단 및 그 조합으로 실현될 것이다.The object of the present invention is not limited to the object mentioned above. The objects of the present invention will become more apparent from the following description, and will be realized by means and combinations thereof described in the claims.
본 발명의 일측면은 가노데릭산(ganoderic acid) 또는 이의 유도체를 유효성분으로 포함하는 피부염증의 예방, 치료 또는 경감용 조성물을 제공한다. One aspect of the present invention provides a composition for preventing, treating or alleviating skin inflammation comprising ganoderic acid or a derivative thereof as an active ingredient.
본 발명의 일측면은 가노데릭산(ganoderic acid) 또는 이의 유도체를 유효성분으로 포함하는 아토피의 예방, 치료 또는 경감용 조성물을 제공한다. One aspect of the present invention provides a composition for preventing, treating, or alleviating atopy comprising ganoderic acid or a derivative thereof as an active ingredient.
본 발명의 일측면은 가노데릭산(ganoderic acid) 또는 이의 유도체를 유효성분으로 포함하는 건선의 예방, 치료 또는 경감용 조성물을 제공한다. One aspect of the present invention provides a composition for preventing, treating or alleviating psoriasis comprising ganoderic acid or a derivative thereof as an active ingredient.
본 발명의 일측면에 있어서, 상기 가노데릭산 또는 이의 유도체는 가노데릭산 eta (GAeta), 부틸 가노데릭산 A (BGAA), 가노데릭산 C2 (GAC2), 가노데릭산 A (GAA), 가노데릭산 G (GAG), 가노드레닉산 B (GDAB), 가노데릭산 B (GAB), 가노드레닉산 K (GDAK), 가노데릭산 K (GAK), 가노드레닉산 D (GDAD), 가노데릭산 D (GAD), 가노데릭산 H (GAH), 가노데릭산 C6 (GAC6) 및 루시데닉산 D 중에서 선택된 어느 하나 이상인, 조성물을 제공한다.In one aspect of the present invention, the ganoderic acid or its derivative is ganoderic acid eta (GAeta), butyl ganoderic acid A (BGAA), ganoderic acid C2 (GAC2), ganoderic acid A (GAA), gano Derric Acid G (GAG), Ganoderenic Acid B (GDAB), Ganoderic Acid B (GAB), Ganoderic Acid K (GDAK), Ganoderic Acid K (GAK), Ganoderenic Acid D (GDAD), Ganoderic Acid D (GAD), Ganoderic acid H (GAH), Ganoderic acid C6 (GAC6) and lucidenic acid D at least one selected from the group consisting of, provides a composition.
본 발명의 일측면에 있어서, 상기 가노데릭산 또는 이의 유도체는 영지버섯(ganoderma lucidum)의 추출물로부터 분리 또는 수득된 것인, 조성물을 제공한다.In one aspect of the present invention, the ganoderic acid or its derivative is isolated or obtained from the extract of reishi mushroom ( ganoderma lucidum ), it provides a composition.
본 발명의 일측면에 있어서, 상기 영지버섯의 추출물은 에탄올 추출물인, 조성물을 제공한다.In one aspect of the present invention, the extract of the reishi mushroom is an ethanol extract, it provides a composition.
본 발명의 일측면에 있어서, 상기 영지버섯의 추출물은 30 내지 80℃의 온도에서, 2 내지 40 시간 동안 추출된 것인, 조성물을 제공한다.In one aspect of the present invention, the extract of the reishi mushroom is extracted at a temperature of 30 to 80 ℃, for 2 to 40 hours, it provides a composition.
본 발명의 일측면에 있어서, 상기 유효성분은 케모카인, 사이토카인 및 S100 칼슘 결합 단백질 중 어느 하나 이상을 억제하는, 조성물을 제공한다.In one aspect of the present invention, the active ingredient provides a composition that inhibits any one or more of chemokines, cytokines and S100 calcium binding protein.
본 발명의 일측면에 있어서, 상기 케모카인은 CXCL8, CCL5 및 CCL20 중 어느 하나 이상이며, 상기 사이토카인은 IL-6 및 IL-23/p19 중 어느 하나 이상이며, 상기 S100 칼슘 결합 단백질은 S100A7, S100A8 및 S100A9 중 어느 하나 이상인, 조성물을 제공한다.In one aspect of the present invention, the chemokine is any one or more of CXCL8, CCL5, and CCL20, the cytokine is any one or more of IL-6 and IL-23/p19, and the S100 calcium binding protein is S100A7, S100A8 And any one or more of S100A9 provides a composition.
본 발명의 일측면에 있어서, 상기 조성물은 약학 조성물인, 조성물을 제공한다.In one aspect of the present invention, the composition provides a pharmaceutical composition, the composition.
본 발명의 일측면에 있어서, 상기 조성물은 화장료 조성물인, 조성물을 제공한다.In one aspect of the present invention, the composition provides a cosmetic composition, the composition.
본 발명의 일측면에 있어서, 상기 조성물은 건강기능 식품 조성물인, 조성물을 제공한다.In one aspect of the present invention, the composition provides a health functional food composition, the composition.
본 발명의 다른 측면은, 피부염증, 아토피 또는 건선의 예방, 치료 또는 경감용 가노데릭산 또는 이의 유도체의 제조 방법에 있어서, 영지버섯(ganoderma lucidum)을 30 내지 80℃의 온도에서, 2 내지 40 시간 동안 에탄올 용매를 이용하여 추출하는 단계를 포함하는, 제조 방법을 제공한다.Another aspect of the present invention, in the method for producing ganoderic acid or a derivative thereof for the prevention, treatment or alleviation of skin inflammation, atopic dermatitis or psoriasis, reishi mushroom ( ganoderma lucidum ) at a temperature of 30 to 80 ℃, 2 to 40 It provides a manufacturing method comprising the step of extracting using an ethanol solvent for a period of time.
본 발명의 다른 측면에 있어서, 상기 추출하는 단계는 초음파 추출인, 제조 방법을 제공한다.In another aspect of the present invention, the extracting step is ultrasonic extraction, provides a manufacturing method.
본 발명의 다른 측면에 있어서, 상기 방법은 추출 후 수득된 조추출물을 재용해하는 단계를 더 포함하는, 제조 방법을 제공한다.In another aspect of the present invention, the method provides a manufacturing method, further comprising the step of re-dissolving the crude extract obtained after extraction.
본 발명의 유효성분 또는 이를 포함하는 조성물은 건선의 예방, 치료 또는 경감효과가 우수하다.The active ingredient of the present invention or a composition comprising the same is excellent in preventing, treating or alleviating psoriasis.
본 발명의 유효성분 또는 이를 포함하는 조성물은 아토피의 예방, 치료 또는 경감효과가 우수하다.The active ingredient of the present invention or a composition comprising the same is excellent in preventing, treating or alleviating atopy.
본 발명의 유효성분 또는 이를 포함하는 조성물은 피부염증의 예방, 치료 또는 경감효과가 우수하다.The active ingredient of the present invention or a composition comprising the same is excellent in preventing, treating or alleviating skin inflammation.
본 발명의 유효성분 또는 이를 포함하는 조성물은 기존 스테로이드계 물질 및 항히스타민제와 달리 부작용 없이 건선, 아토피, 피부염증의 예방, 치료 또는 경감 효과가 우수하다.The active ingredient of the present invention or a composition comprising the same is excellent in preventing, treating or alleviating psoriasis, atopic dermatitis, and skin inflammation without side effects, unlike conventional steroid-based substances and antihistamines.
본 발명의 유효성분 또는 이를 포함하는 조성물은 피부 각질 세포인 HeCaT 세포에서 염증을 유발하는 인자들인 염증성 사이토카인들 (IL-6, S100A7, S100A8, S100A9, CCL5, CCL20, CXCL8)의 발현량이 mRNA 수준에서 현저히 감소시킬 수 있다.The active ingredient of the present invention or a composition comprising the same is expressed in mRNA levels of inflammatory cytokines (IL-6, S100A7, S100A8, S100A9, CCL5, CCL20, CXCL8), which are factors that induce inflammation in HeCaT cells, which are keratinocytes of the skin. can be significantly reduced in
본 발명의 유효성분인 가노데릭산 또는 이의 유도체의 제조 방법은 높은 수율로 상기 유효성분을 수득할 수 있다.The method for producing ganoderic acid or a derivative thereof, which is the active ingredient of the present invention, can obtain the active ingredient in high yield.
본 발명의 효과는 이상에서 언급한 효과로 한정되지 않는다. 본 발명의 효과는 이하의 설명에서 추론 가능한 모든 효과를 포함하는 것으로 이해되어야 할 것이다. The effects of the present invention are not limited to the above-mentioned effects. It should be understood that the effects of the present invention include all effects that can be inferred from the following description.
도 1은 본원발명 실시예 1의 영지버섯 추출물 및 그 분획물의 제조 과정을 그린 도면이다.
도 2는 본원발명 실시예 1의 영지버섯 추출물의 성분을 실험예 1에 따라 UPLC-QTOF-MS/MS 로 분석한 결과이다.
도 3은 본원발명 실시예 1의 반정량 분석 결과이다.
도 4(도 4a 내지 도 4c)은 본원발명 실험예 2에 따른 세포 독성 확인 결과이다.
도 5는 본원발명 실험예 3에서 각 추출조건에 따른 추출물의 IL-6 유전자 발현 억제 결과이다.
도 6는 본원발명 실험예 3에서 각 추출조건에 따른 추출물의 S100A7 유전자 발현 억제 결과이다.
도 7(도 7a 내지 7c)은 40℃ 12시간, 55℃ 12시간, 24시간, 70℃ 1시간의 특정 조건에서 추출한 추출물의 피부염증활성 관련 유전자의 발현활성을 측정한 결과이다.1 is a diagram illustrating a process for preparing a reishi mushroom extract and a fraction thereof of Example 1 of the present invention.
2 is a result of analyzing the components of the reishi mushroom extract of Example 1 of the present invention by UPLC-QTOF-MS/MS according to Experimental Example 1.
3 is a semi-quantitative analysis result of Example 1 of the present invention.
4 ( FIGS. 4a to 4c ) is a cytotoxicity confirmation result according to Experimental Example 2 of the present invention.
5 is a result of suppression of IL-6 gene expression of the extract according to each extraction condition in Experimental Example 3 of the present invention.
6 is a result of S100A7 gene expression inhibition of the extract according to each extraction condition in Experimental Example 3 of the present invention.
7 ( FIGS. 7a to 7c ) is a result of measuring the expression activity of genes related to skin inflammatory activity of extracts extracted under specific conditions of 40° C. for 12 hours, 55° C. for 12 hours, 24 hours, and 70° C. for 1 hour.
달리 명시되지 않는 한, 본 명세서에서 사용된 성분, 반응 조건, 성분의 함량을 표현하는 모든 숫자, 값 및/또는 표현은, 이러한 숫자들이 본질적으로 다른 것들 중에서 이러한 값을 얻는 데 발생하는 측정의 다양한 불확실성이 반영된 근사치들이므로, 모든 경우 "약"이라는 용어에 의해 수식되는 것으로 이해되어야 한다. 또한, 본 기재에서 수치범위가 개시되는 경우, 이러한 범위는 연속적이며, 달리 지적되지 않는 한 이러한 범위의 최소값으로부터 최대값이 포함된 상기 최대값까지의 모든 값을 포함한다. 더 나아가, 이러한 범위가 정수를 지칭하는 경우, 달리 지적되지 않는 한 최소값으로부터 최대값이 포함된 상기 최대값까지를 포함하는 모든 정수가 포함된다.Unless otherwise specified, all numbers, values, and/or expressions expressing ingredients, reaction conditions, and amounts of ingredients used herein refer to a variety of measures that may occur in obtaining such values, among others, in which such numbers are inherently different. Since they are approximations reflecting uncertainty, it should be understood as being modified by the term "about" in all cases. Also, where numerical ranges are disclosed in this description, such ranges are continuous and inclusive of all values from the minimum to the maximum inclusive of the range, unless otherwise indicated. Furthermore, when such ranges refer to integers, all integers inclusive from the minimum to the maximum inclusive are included, unless otherwise indicated.
본 명세서에 있어서, 범위가 변수에 대해 기재되는 경우, 상기 변수는 상기 범위의 기재된 종료점들을 포함하는 기재된 범위 내의 모든 값들을 포함하는 것으로 이해될 것이다. 예를 들면, "5 내지 10"의 범위는 5, 6, 7, 8, 9, 및 10의 값들뿐만 아니라 6 내지 10, 7 내지 10, 6 내지 9, 7 내지 9 등의 임의의 하위 범위를 포함하고, 5.5, 6.5, 7.5, 5.5 내지 8.5 및 6.5 내지 9 등과 같은 기재된 범위의 범주에 타당한 정수들 사이의 임의의 값도 포함하는 것으로 이해될 것이다. 또한 예를 들면, "10% 내지 30%"의 범위는 10%, 11%, 12%, 13% 등의 값들과 30%까지를 포함하는 모든 정수들뿐만 아니라 10% 내지 15%, 12% 내지 18%, 20% 내지 30% 등의 임의의 하위 범위를 포함하고, 10.5%, 15.5%, 25.5% 등과 같이 기재된 범위의 범주 내의 타당한 정수들 사이의 임의의 값도 포함하는 것으로 이해될 것이다.In this specification, when a range is described for a variable, the variable will be understood to include all values within the stated range including the stated endpoints of the range. For example, a range of “5 to 10” includes the values of 5, 6, 7, 8, 9, and 10, as well as any subranges such as 6 to 10, 7 to 10, 6 to 9, 7 to 9, etc. It will be understood to include any value between integers that are appropriate for the scope of the recited range, such as 5.5, 6.5, 7.5, 5.5 to 8.5 and 6.5 to 9, and the like. Also for example, a range of "10% to 30%" includes values such as 10%, 11%, 12%, 13%, and all integers up to and including 30%, as well as 10% to 15%, 12% to It will be understood to include any subrange, such as 18%, 20% to 30%, etc., as well as any value between reasonable integers within the scope of the recited range, such as 10.5%, 15.5%, 25.5%, and the like.
이하, 본 발명에 대하여 상세히 설명한다.Hereinafter, the present invention will be described in detail.
가노데릭산은 3개의 사이클로헥산 링과 한 개의 사이클로펜탄 링으로 구성된 대표적인 트리터페노이드계 성분으로 항산화, 항당뇨, 항비만 및 항염 등의 생리활성뿐 아니라 면역 기능을 증진시키고, 특히 항염증 활성이 우수한 성분이다. 영지버섯 (Ganoderma lucidum)의 주요 기능성 성분인 가노데릭산은 지금까지 약 120여개의 성분들이 보고되어 왔으며, 기존의 연구들에 따르면, 탄소 번호 3, 7, 11, 12, 15번에 연결되어 있는 하이드록실기, 카르보닐기, 아세틸기 등의 작용기의 종류와 여부에 따라 생리 활성이 다르게 나타나는 것으로 알려져 있다. 그러나 가노데릭산이 어떤 조건에서 피부 염증에 효능이 있는지에 대해 보고된 바가 없다.Ganoderic acid is a representative triterpenoid component composed of three cyclohexane rings and one cyclopentane ring. is an ingredient Reishi mushroom ( Ganoderma lucidum ) Ganoderic acid, the main functional ingredient, has been reported about 120 ingredients so far, and according to previous studies, the hydride linked to
본 특허는 피부염증 효과에 탁월한 영지버섯(ganoderma lucidum) 추출물 및 추출 방법에 관한 것으로, 본 발명의 영지버섯 추출물은 피부 각질 세포인 HeCaT 세포에서 염증을 유발하는 인자들인 염증성 사이토카인들 (IL-6, S100A7, S100A8, S100A9, CCL5, CCL20, CXCL8)의 발현량이 mRNA 수준에서 감소한 것을 확인하였다.This patent relates to a ganoderma lucidum extract and an extraction method excellent for skin inflammatory effects, and the reishi mushroom extract of the present invention contains inflammatory cytokines (IL-6 , S100A7, S100A8, S100A9, CCL5, CCL20, CXCL8) was confirmed to be reduced at the mRNA level.
기존의 합성 항염증제인 스테로이드제의 경우 구역질, 소화불량 등의 부작용을 유발해 천연 항염증제에 대한 관심과 수요가 높아지고 있다. 이에 본원발명의 발명자들은 영지버섯의 항염증 활성 성분인 가노데릭산을 고함유하고 항염증 활성이 증진된 영지버섯 추출물 제조법을 발명하여, 높은 함량의 가노데릭산을 포함하는 조성물을 제공할 수 있다.Steroids, which are conventional synthetic anti-inflammatory drugs, cause side effects such as nausea and indigestion, so interest and demand for natural anti-inflammatory drugs are increasing. Accordingly, the inventors of the present invention invent a method for preparing a reishi mushroom extract containing a high content of ganoderic acid, which is an anti-inflammatory active ingredient of reishi mushroom, and having enhanced anti-inflammatory activity, thereby providing a composition containing a high content of ganoderic acid. .
이하, 본 발명의 다양한 측면에 대하여 설명한다.Hereinafter, various aspects of the present invention will be described.
본 발명의 일측면은 가노데릭산(ganoderic acid) 또는 이의 유도체를 유효성분으로 포함하는 피부염증의 예방, 치료 또는 경감용 조성물을 제공한다. One aspect of the present invention provides a composition for preventing, treating or alleviating skin inflammation comprising ganoderic acid or a derivative thereof as an active ingredient.
본 발명의 일측면은 가노데릭산(ganoderic acid) 또는 이의 유도체를 유효성분으로 포함하는 아토피의 예방, 치료 또는 경감용 조성물을 제공한다. One aspect of the present invention provides a composition for preventing, treating, or alleviating atopy comprising ganoderic acid or a derivative thereof as an active ingredient.
본 발명의 일측면은 가노데릭산(ganoderic acid) 또는 이의 유도체를 유효성분으로 포함하는 건선의 예방, 치료 또는 경감용 조성물을 제공한다. One aspect of the present invention provides a composition for preventing, treating or alleviating psoriasis comprising ganoderic acid or a derivative thereof as an active ingredient.
본 발명의 일측면에 있어서, 상기 유효성분은 0.1 중량% 내지 90 중량%로 포함되는 조성물을 제공한다.In one aspect of the present invention, the active ingredient provides a composition comprising 0.1 wt% to 90 wt%.
본 발명의 일측면에 있어서, 상기 가노데릭산 또는 이의 유도체는 가노데릭산 eta (GAeta), 부틸 가노데릭산 A (BGAA), 가노데릭산 C2 (GAC2), 가노데릭산 A (GAA), 가노데릭산 G (GAG), 가노드레닉산 B (GDAB), 가노데릭산 B (GAB), 가노드레닉산 K (GDAK), 가노데릭산 K (GAK), 가노드레닉산 D (GDAD), 가노데릭산 D (GAD), 가노데릭산 H (GAH), 가노데릭산 C6 (GAC6) 및 루시데닉산 D 중에서 선택된 어느 하나 이상인, 조성물을 제공한다.In one aspect of the present invention, the ganoderic acid or its derivative is ganoderic acid eta (GAeta), butyl ganoderic acid A (BGAA), ganoderic acid C2 (GAC2), ganoderic acid A (GAA), gano Derric Acid G (GAG), Ganoderenic Acid B (GDAB), Ganoderic Acid B (GAB), Ganoderic Acid K (GDAK), Ganoderic Acid K (GAK), Ganoderenic Acid D (GDAD), Ganoderic Acid D (GAD), Ganoderic acid H (GAH), Ganoderic acid C6 (GAC6) and lucidenic acid D at least one selected from the group consisting of, provides a composition.
본 발명의 일측면에 있어서, 상기 가노데릭산 또는 이의 유도체는 영지버섯(ganoderma lucidum)의 추출물로부터 분리 또는 수득된 것인, 조성물을 제공한다.In one aspect of the present invention, the ganoderic acid or its derivative is isolated or obtained from the extract of reishi mushroom ( ganoderma lucidum ), it provides a composition.
본 명세서에서 "추출물"이라 함은, 천연물로부터 그 안의 성분을 뽑아냄으로써 얻어진 물질이라면, 뽑아내는 방 법이나 성분의 종류와 무관하게 모두 포함한다. 예컨대, 물이나 유기 용매를 이용하여 천연물로부터 용매에 용 해되는 성분을 추출해 낸 것, 천연물의 특정 성분만을 추출하여 얻어진 것 등을 모두 포함하는 광의의 개념이다. 본 발명의 일구현예에서, 상기 유기 용매는, 특별히 제한되는 것은 아니며, 메탄올, 에탄올, 이소프 로필알코올, n-프로필 알코올, n-부탄올 및 이소부탄올 등의 C1~C5의 저급 알코올, 글리세롤, 에틸렌글리콜, 프 로필렌글리콜, 1,3-부틸 렌글리콜 등의 다가 알코올, 메틸아세테이트, 에틸아세테이트, 벤젠, n-헥산, 디에틸에 테르, 디클로로메탄, 클로로포름 등의 탄화수소계 용매, 그리고 석유에테르, 메틸아세테이트, 벤젠, 헥산, 클로 로포름, 메틸렌클로라이드, 디메틸에테르, 에틸아세테이트 등의 비극성 유기용매 등 일 수 있다.As used herein, the term "extract" includes all substances obtained by extracting the components therein from natural products, regardless of the extraction method or the type of components. For example, it is a broad concept that includes both extracting a component soluble in a solvent from a natural product using water or an organic solvent, and extracting only a specific component of a natural product. In one embodiment of the present invention, the organic solvent is not particularly limited, and C1-C5 lower alcohols such as methanol, ethanol, isopropyl alcohol, n-propyl alcohol, n-butanol and isobutanol, glycerol, and ethylene Polyhydric alcohols such as glycol, propylene glycol, 1,3-butylene glycol, hydrocarbon solvents such as methyl acetate, ethyl acetate, benzene, n-hexane, diethyl ether, dichloromethane, chloroform, petroleum ether, It may be a non-polar organic solvent such as methyl acetate, benzene, hexane, chloroform, methylene chloride, dimethyl ether, and ethyl acetate.
본 발명의 일측면에 있어서, 상기 추출물은 물, C1-C5 알코올, 아세톤, 아세톤 수용액 또는 C1-C5 알코올 수용액을 이용하여 추출한 추출물인, 조성물을 제공한다.In one aspect of the present invention, the extract is an extract extracted using water, C 1 -C 5 alcohol, acetone, an aqueous acetone solution or a C 1 -C 5 aqueous alcohol solution, it provides a composition.
본 발명의 일측면에서, 상기 C1-C5 알코올은, 메탄올, 에탄올, 이소프로필알코올, n-프로필알코올, n-부탄올 및 이소부탄올로 구성된 군으로부터 선택되는 어느 하나 이상을 이용한, 조성물을 제공한다.In one aspect of the present invention, the C1-C5 alcohol is methanol, ethanol, isopropyl alcohol, n-propyl alcohol, n-butanol and provides a composition using any one or more selected from the group consisting of isobutanol.
본 발명의 일측면에 있어서, 상기 C1-C5 알코올 수용액 및 아세톤 수용액의 농도는 각각 독립적으로 10% 내지 90% (v/v)인, 조성물을 제공한다.In one aspect of the present invention, the concentration of the C 1 -C 5 aqueous alcohol solution and the aqueous acetone solution is each independently 10% to 90% (v/v), it provides a composition.
본 발명의 일측면에서, 상기 유효성분은 조성물 전체 중량을 기준으로 0.001 내지 90 중량%로 포함될 수 있다. 일 구현예에서, 상기 유효성분은 조성물 전체 중량을 기준으로 0.001 중량% 이상, 0.01중량% 이상, 0.1중량% 이상, 1중량% 이상, 1.1 중량% 이상, 1.5 중량% 이상, 2 중량% 이상, 3 중량% 이상, 5 중량% 이상, 10 중량% 이상, 20중량% 이상 또는 30 중량% 이상일 수 있다. 또한, 상기 인삼 꽃대 추출물은 조성물 전 체 중량을 기준으로 90 중량% 이하, 85중량% 이하, 80중량% 이하, 70중량% 이하, 50중량% 이하, 40중량% 이하, 30중량% 이하, 20중량% 이하, 10 중량% 이하, 5 중량% 이하, 4 중량% 이하, 3 중량% 이하, 2 중량% 이하, 1 중 량% 이하, 0.1 중량% 이하 또는 0.05중량% 이하일 수 있다.In one aspect of the present invention, the active ingredient may be included in an amount of 0.001 to 90% by weight based on the total weight of the composition. In one embodiment, the active ingredient is 0.001% by weight or more, 0.01% by weight or more, 0.1% by weight or more, 1% by weight or more, 1.1% by weight or more, 1.5% by weight or more, 2% by weight or more, based on the total weight of the composition, 3 wt% or more, 5 wt% or more, 10 wt% or more, 20 wt% or more, or 30 wt% or more. In addition, the ginseng flower stem extract is 90% by weight or less, 85% by weight or less, 80% by weight or less, 70% by weight or less, 50% by weight or less, 40% by weight or less, 30% by weight or less, 20% by weight or less, based on the total weight of the composition weight % or less, 10 wt% or less, 5 wt% or less, 4 wt% or less, 3 wt% or less, 2 wt% or less, 1 wt% or less, 0.1 wt% or less, or 0.05 wt% or less.
본 발명의 일측면에 있어서, 상기 영지버섯의 추출물은 에탄올 추출물인, 조성물을 제공한다.In one aspect of the present invention, the extract of the reishi mushroom is an ethanol extract, it provides a composition.
본 발명의 일측면에 있어서, 상기 영지버섯의 추출물은 30 내지 80℃의 온도에서, 2 내지 40 시간 동안 추출된 것인, 조성물을 제공한다.In one aspect of the present invention, the extract of the reishi mushroom is extracted at a temperature of 30 to 80 ℃, for 2 to 40 hours, it provides a composition.
본 발명의 일측면에 있어서, 상기 영지버섯의 추출물은 30 내지 60℃의 온도에서, 2 내지 15 시간 동안 추출된 것인, 조성물을 제공한다.In one aspect of the present invention, the extract of the reishi mushroom at a temperature of 30 to 60 ℃, it provides a composition that will be extracted for 2 to 15 hours.
본 발명의 일측면에 있어서, 상기 영지버섯의 추출물은 35 내지 45℃의 온도에서, 10 내지 14 시간 동안 추출된 것인, 조성물을 제공한다.In one aspect of the present invention, the extract of the reishi mushroom is extracted at a temperature of 35 to 45 ℃, for 10 to 14 hours, it provides a composition.
본 발명의 일측면에 있어서, 상기 영지버섯의 추출물은 50 내지 60℃의 온도에서, 10 내지 14 시간 동안 추출된 것인, 조성물을 제공한다.In one aspect of the present invention, the extract of the reishi mushroom at a temperature of 50 to 60 ℃, it provides a composition that is extracted for 10 to 14 hours.
본 발명의 일측면에 있어서, 상기 유효성분은 케모카인, 사이토카인 및 S100 칼슘 결합 단백질 중 어느 하나 이상을 억제하는, 조성물을 제공한다.In one aspect of the present invention, the active ingredient provides a composition that inhibits any one or more of chemokines, cytokines and S100 calcium binding protein.
본 발명의 일측면에 있어서, 상기 케모카인은 CXCL8, CCL5 및 CCL20 중 어느 하나 이상이며, 상기 사이토카인은 IL-6 및 IL-23/p19 중 어느 하나 이상이며, 상기 S100 칼슘 결합 단백질은 S100A7, S100A8 및 S100A9 중 어느 하나 이상인, 조성물을 제공한다.In one aspect of the present invention, the chemokine is any one or more of CXCL8, CCL5, and CCL20, the cytokine is any one or more of IL-6 and IL-23/p19, and the S100 calcium binding protein is S100A7, S100A8 And any one or more of S100A9 provides a composition.
본 발명의 일측면에 있어서, 상기 조성물은 약학 조성물인, 조성물을 제공한다. 본 발명의 약학 조성물은 약제의 제조에 통상적으로 사용하는 적절한 담체, 부형제 및/또는 희석제를 더 포함하여 경구 또는 비경구 투여에 적합한 의약품 제제의 형태로 제조가 가능하다. 또한, 본 발명의 약학 조성물을 사용하여 통상적인 방법에 따라 약학 제형을 제조할 수 있다. 제형의 제조에 있어, 활성성분을 담체와 함께 혼합하거나, 담체로 희석하거나, 캅셀, 새세이 또는 기타 용기 형태의 담체 내에 봉입시킬 수 있다. 따라서 제형은 정제, 환제, 분제, 캡슐, 새세이, 엘릭씨르, 현탁제, 에멀젼, 액제, 시럽제, 에어로졸, 연질 또는 경질 젤라틴 캅셀제, 주사용 용액 또는 현탁액, 연고제, 크림제, 겔제 또는 로숀제 등의 형태일 수 있다. In one aspect of the present invention, the composition provides a pharmaceutical composition, the composition. The pharmaceutical composition of the present invention can be prepared in the form of a pharmaceutical formulation suitable for oral or parenteral administration by further including an appropriate carrier, excipient and/or diluent commonly used in the manufacture of pharmaceuticals. In addition, a pharmaceutical formulation can be prepared according to a conventional method using the pharmaceutical composition of the present invention. In preparing the formulation, the active ingredient may be mixed with a carrier, diluted with a carrier, or enclosed in a carrier in the form of a capsule, sachet or other container. Accordingly, the dosage form is a tablet, pill, powder, capsule, sachet, elixir, suspension, emulsion, solution, syrup, aerosol, soft or hard gelatin capsule, solution or suspension for injection, ointment, cream, gel or lotion. It may be in the form of
본 발명의 약학 조성물에 포함될 수 있는 적합한 담체, 부형제 및 희석제는 예를 들면 락토즈, 덱스트로즈, 수크로즈, 솔비톨, 만니톨, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로즈, 폴리비닐피롤리돈, 물, 메틸하이드록시벤조에이트, 프로필하이드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다. 또한, 제형 제조 시에 통상적으로 사용되고 있는 충진제, 항응집제, 윤활제, 습윤제, 향료, 유화제, 방부제 등을 추가로 포함할 수 있다. 본 발명의 약제 조성물은 포유동물에 투여된 후 활성성분의 신속, 지속 또는 지연된 방출을 제공할 수 있도록 당 업계에 잘 알려진 방법을 사용하여 제형화 될 수 있다. Suitable carriers, excipients and diluents that may be included in the pharmaceutical composition of the present invention are, for example, lactose, dextrose, sucrose, sorbitol, mannitol, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone. , water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition, fillers, anti-aggregants, lubricants, wetting agents, fragrances, emulsifiers, preservatives, and the like, which are commonly used in the preparation of formulations may be further included. The pharmaceutical composition of the present invention may be formulated using methods well known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal.
본 발명의 약학 조성물의 투여량은 환자의 상태 및 체중, 질병의 정도, 약물형태, 투여경로 및 기간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다. 환자의 몸무게 대비 활성성분은 0.001 ㎎/㎏ 내지 500 ㎎/㎏ 범위일 수 있고, 바람직하게는 0.001 내지 200 ㎎/㎏으로 투여하는 것이 좋다. 투여는 하루에 한번 투여할 수도 있고, 또는 수회 나누어 투여할 수도 있다. 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다.The dosage of the pharmaceutical composition of the present invention varies depending on the condition and weight of the patient, the degree of disease, the drug form, the route and duration of administration, but may be appropriately selected by those skilled in the art. The active ingredient relative to the body weight of the patient may be in the range of 0.001 mg/kg to 500 mg/kg, and it is preferably administered at 0.001 to 200 mg/kg. Administration may be administered once a day, or may be administered in several divided doses. The above dosage does not limit the scope of the present invention in any way.
본 발명의 일측면에 있어서, 상기 조성물은 화장료 조성물인, 조성물을 제공한다.In one aspect of the present invention, the composition provides a cosmetic composition, the composition.
상기 화장품 조성물은 제형이 특별히 한정되지 않으며, 목적하는 바에 따라 적절히 선택할 수 있다. 예를 들어, 스킨로션, 스킨소프너, 스킨토너, 아스트린젠트, 로션, 밀크로션, 모이스쳐 로션, 영양로션, 맛사지크림, 영양크림, 모이스처크림, 핸드크림, 파운데이션, 에센스, 영양에센스, 팩, 비누, 클렌징폼, 클렌징로션, 클렌징크림, 바디로션 및 바디클린저로 이루어진 군으로부터 선택된 어느 하나 이상의 제형으로 제조될 수 있으나, 이에 제한되는 것은 아니다.The formulation of the cosmetic composition is not particularly limited and may be appropriately selected according to the purpose. For example, skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, hand cream, foundation, essence, nourishing essence, pack, soap, cleansing It may be prepared in any one or more formulations selected from the group consisting of foam, cleansing lotion, cleansing cream, body lotion and body cleanser, but is not limited thereto.
본 발명의 제형이 페이스트, 크림 또는 겔인 경우에는 담체 성분으로서 동물섬유, 식물섬유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산화아연 등이 이용될 수 있다.When the formulation of the present invention is a paste, cream or gel, animal fiber, vegetable fiber, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component. can
본 발명의 제형이 파우더 또는 스프레이인 경우에는 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄히드록시드, 칼슘 실리케이트 또는 폴리아미드 파우더가 이용될 수 있고, 특히 스프레이인 경우에는 추가적으로 클로로플루오로히드로카본, 프로판/부탄 또는 디메틸 에테르와 같은 추진체를 포함할 수 있다.When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and in particular, in the case of a spray, additional chlorofluorohydrocarbon, propane /may contain propellants such as butane or dimethyl ether.
본 발명의 제형이 용액 또는 유탁액의 경우에는 담체 성분으로서 용매, 용매화제 또는 유탁화제가 이용되고, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌 글리콜, 1,3-부틸글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜 또는 소르비탄의 지방산 에스테르가 있다.When the formulation of the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butylglycol oil, glycerol fatty esters, fatty acid esters of polyethylene glycol or sorbitan.
본 발명의 제형이 현탁액인 경우에는 담체 성분으로서 물, 에탄올 또는 프로필렌 글리콜과 같은 액상 희석제, 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르 및 폴리옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미소결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가 또는 트라칸트 등이 이용될 수 있다.When the formulation of the present invention is a suspension, as a carrier component, water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystalline Cellulose, aluminum metahydroxide, bentonite, agar or tracanth may be used.
본 발명의 제형이 계면-활성제 함유 클린징인 경우에는 담체 성분으로서 지방족 알코올 설페이트, 지방족 알코올 에테르 설페이트, 설포숙신산 모노에스테르, 이세티오네이트, 이미다졸리늄 유도체, 메틸타우레이트, 사르코시네이트, 지방산 아미드 에테르 설페이트, 알킬아미도베타인, 지방족 알코올, 지방산 글리세리드, 지방산 디에탄올아미드, 식물성 유, 리놀린 유도체 또는 에톡실화 글리세롤 지방산 에스테르 등이 이용될 수 있다.When the formulation of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide as carrier components Ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linolin derivative or ethoxylated glycerol fatty acid ester and the like can be used.
상기 유효성분의 함량은 특별히 제한되지 않으나, 조성물 총 중량을 기준으로 0.001 내지 100 중량%로 포함될 수 있다. 상기 유효 성분이 상기 함량을 만족하는 경우 부작용 없이 우수한 효능을 나타낼 수 있다.The content of the active ingredient is not particularly limited, but may be included in an amount of 0.001 to 100% by weight based on the total weight of the composition. When the active ingredient satisfies the above content, excellent efficacy may be exhibited without side effects.
상기 화장품 조성물에는 기능성 첨가물 및 일반적인 화장품 조성물에 포함되는 성분이 추가로 포함될 수 있다. 상기 기능성 첨가물로는 수용성 비타민, 유용성 비타민, 고분자 펩티드, 고분자 다당, 스핑고 지질 및 해초 엑기스로 이루어진 군에서 선택된 성분을 포함할 수 있다.The cosmetic composition may further include functional additives and components included in general cosmetic compositions. The functional additive may include a component selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high-molecular peptides, high-molecular polysaccharides, sphingolipids, and seaweed extract.
본 발명의 화장품 조성물에는 또한, 상기 기능성 첨가물과 더불어 필요에 따라 일반적인 화장품 조성물에 포함되는 성분을 배합해도 된다. 이외에 포함되는 배합 성분으로서는 유지 성분, 보습제, 에몰리엔트제, 계면 활성제, 유기 및 무기 안료, 유기 분체, 자외선 흡수제, 방부제, 살균제, 산화 방지제, 식물 추출물, pH 조정제, 알콜, 색소, 향료, 혈행 촉진제, 냉감제, 제한(制汗)제, 정제수 등을 들 수 있다.In the cosmetic composition of the present invention, a component contained in a general cosmetic composition may be further blended with the functional additive as needed. Other ingredients included include oils and fats, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, UV absorbers, preservatives, fungicides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, fragrances, blood circulation An accelerator, a cooling agent, a limiting agent, purified water, etc. are mentioned.
본 발명의 일측면에 있어서, 상기 조성물은 건강기능 식품 조성물인, 조성물을 제공한다.In one aspect of the present invention, the composition provides a health functional food composition, the composition.
본 발명의 건강식품 조성물은 상기 유효성분을 포함하는 것으로, 그 종류에는 특별한 제한은 없다. 상기 식품의 예로는 드링크제, 육류, 소시지, 빵, 비스킷, 떡, 선식, 초콜릿, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 알코올 음료 및 비타민 복합제, 유제품 및 유가공 제품 등이 있으며, 그 밖에도 통상적인 의미에서의 건강기능식품을 모두 포함한다. 활성성분으로서 영지버섯 추출물 또는 이로부터 분획한 용매분획물은 식품에 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효 함량은 사용 목적(예방 또는 개선용)에 따라 적합하게 결정될 수 있으며, 건강식품의 전체 중량대비 0.001 내지 70 중량% 범위로 포함될 수 있다. 그러나 건강 및 위생을 목적으로 하거나 또는 건강 조절을 목적으로 하는 장기간 섭취의 경우에는 상기 양은 상기 범위 이하일 수 있으며, 안전성 면에서 아무런 문제가 없기 때문에 유효성분은 상기 범위 이상의 양으로도 사용될 수 있다. 예를 들면, 건강음료로 제조하는 경우는 상기 활성성분 이외에도 음료 제조 시에 통상적으로 사용되는 첨가제로서 천연 탄수화물 또는 향미제 등을 함유할 수 있다. 상기 천연 탄수화물은 모노사카라이드(예를 들어, 포도당, 과당 등), 디사카라이드(예를 들어, 말토스, 슈크로스 등) 및 폴리사카라이드(예를 들어 덱스트린, 시클로덱스트린 등)와 같은 통상적인 당 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이 포함될 수 있다. 상기 천연 탄수화물은 건강식품의 전체 중량대비 1 내지 20 중량%, 바람직하게는 5 내지 10 중량% 범위로 함유될 수 있다. 상기 향미제는 천연 향미제(타우마틴, 스테비아 추출물, 레바우디오시드 A, 글리시르히진 등) 및 합성 향미제(사카린, 아스파르탐 등)가 포함될 수 있다. 그 밖에도 여러 가지 영양제, 비타민, 광물(전해질), 풍미제(합성 또는 천연 풍미제), 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 또한, 천연 과일 쥬스 및 과일 쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 첨가제의 함량에 특별한 제한을 두고 있지는 않지만, 건강식품의 전체 중량대비 0.1 내지 20 중량%의 범위로 포함될 수 있다.The health food composition of the present invention includes the active ingredient, and there is no particular limitation on the type thereof. Examples of the food include drinks, meat, sausage, bread, biscuits, rice cakes, sun food, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, alcoholic beverages and vitamin complexes, dairy products, and dairy products. As an active ingredient, the reishi mushroom extract or a solvent fraction fractionated therefrom may be added to food as it is or used together with other foods or food ingredients, and may be appropriately used according to a conventional method. The effective content may be appropriately determined depending on the purpose of use (for prevention or improvement), and may be included in the range of 0.001 to 70% by weight based on the total weight of the health food. However, in the case of long-term intake for health and hygiene or health control, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount greater than the above range. For example, in the case of preparing a health drink, natural carbohydrates or flavoring agents may be included as additives commonly used in the manufacture of beverages in addition to the active ingredient. The natural carbohydrates are conventional such as monosaccharides (eg glucose, fructose, etc.), disaccharides (eg maltose, sucrose, etc.) and polysaccharides (eg dextrin, cyclodextrin, etc.). Phosphorus sugar and sugar alcohols such as xylitol, sorbitol, and erythritol may be included. The natural carbohydrate may be contained in an amount of 1 to 20% by weight, preferably 5 to 10% by weight, based on the total weight of the health food. The flavoring agent may include natural flavoring agents (taumatine, stevia extract, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.). In addition, various nutrients, vitamins, minerals (electrolytes), flavoring agents (synthetic or natural flavoring agents), coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives , glycerin, alcohol, a carbonation agent used in carbonated beverages, and the like. It may also contain fruit flesh for the production of natural fruit juice and fruit juice beverages and vegetable beverages. Although there is no particular limitation on the content of these additives, it may be included in the range of 0.1 to 20% by weight based on the total weight of the health food.
본 발명의 다른 측면은, 피부염증, 아토피 또는 건선의 예방, 치료 또는 경감용 가노데릭산 또는 이의 유도체의 제조 방법에 있어서, 영지버섯(ganoderma lucidum)을 30 내지 80℃의 온도에서, 2 내지 40 시간 동안 에탄올 용매를 이용하여 추출하는 단계를 포함하는, 제조 방법을 제공한다.Another aspect of the present invention, in the method for producing ganoderic acid or a derivative thereof for the prevention, treatment or alleviation of skin inflammation, atopic dermatitis or psoriasis, reishi mushroom ( ganoderma lucidum ) at a temperature of 30 to 80 ℃, 2 to 40 It provides a manufacturing method comprising the step of extracting using an ethanol solvent for a period of time.
본 발명의 다른 측면에 있어서, 상기 추출하는 단계는 초음파 추출인, 제조 방법을 제공한다.In another aspect of the present invention, the extracting step is ultrasonic extraction, provides a manufacturing method.
본 발명의 다른 측면에 있어서, 상기 방법은 추출 후 수득된 조추출물을 재용해하는 단계를 더 포함하는, 제조 방법을 제공한다.In another aspect of the present invention, the method provides a manufacturing method, further comprising the step of re-dissolving the crude extract obtained after extraction.
이하, 구체적인 실시예를 통해 본 발명을 보다 구체적으로 설명한다. 하기 실시예는 본 발명의 이해를 돕기 위한 예시에 불과하며, 본 발명의 범위가 이에 한정되는 것은 아니다.Hereinafter, the present invention will be described in more detail through specific examples. The following examples are merely examples to help the understanding of the present invention, and the scope of the present invention is not limited thereto.
실시예Example
실시예 1: 영지버섯 추출물 제조Example 1: Preparation of reishi mushroom extract
본 실시예에서는 영지버섯(Ganoderma lucidum)을 이용해 에탄올로 초음파추출기를 통해 추출하였다. 구체적으로 하기와 같이 영지버섯 추출물을 제조하였다.In this example, reishi mushroom ( Ganoderma lucidum ) was extracted with ethanol through an ultrasonic extractor. Specifically, a reishi mushroom extract was prepared as follows.
영지버섯의 자실체를 열건조시킨 다음 영지버섯 약 2g을 100 % 에탄올에 개의 온도 조건 (40, 55 및 70℃)에서 다양한 시간(1, 3, 6, 12 및 24시간) 초음파 추출기를 이용해 추출하였다. 용액을 거름종이로 여과하여 얻어진 여과액(Filtrate)을 질소 농축기를 통해 농축하여 조 추출물 약 0.2g을 얻었다. 도 1은 영지버섯 추출물 및 그 분획물의 제조 과정을 그린 도면이다. After drying the fruiting body of the reishi mushroom, about 2 g of the reishi mushroom was extracted in 100% ethanol using an ultrasonic extractor at various times (1, 3, 6, 12 and 24 hours) under dog temperature conditions (40, 55 and 70 ° C). . The filtrate obtained by filtering the solution through a filter paper was concentrated through a nitrogen concentrator to obtain about 0.2 g of a crude extract. 1 is a diagram illustrating a manufacturing process of a reishi mushroom extract and a fraction thereof.
[실험예][Experimental example]
실험예 1: 가노데릭산 함량 분석Experimental Example 1: Ganoderic acid content analysis
실시예 1에서 제조된 조추출물은 DMSO를 이용해 재용해하였으며, UPLC-QTOF-MS/MS 분석을 위해 분석용 0.2 ㎛ 멤브레인 필터를 통해 한 번 더 필터링하였다. The crude extract prepared in Example 1 was redissolved using DMSO and filtered once more through a 0.2 μm membrane filter for analysis for UPLC-QTOF-MS/MS analysis.
(1) 함량분석(1) Content analysis
가노데릭산의 분석은 에이질런트의 UPLC 1290 infinity 시리즈/6470 Triple quadrupole MS를 연결하여 진행하였으며, 분석 조건은 표 1과 같았다. Ganoderic acid was analyzed by connecting Agilent's UPLC 1290 infinity series/6470 triple quadrupole MS, and the analysis conditions were as shown in Table 1.
(B) acetonitrile containing 0.2% formic acid(A) distilled water containing 0.2% formic acid
(B) acetonitrile containing 0.2% formic acid
(2) 분석 결과(2) Analysis result
UPLC-QTOF-MS/MS 분석 결과, 도 2에 표시된 바와 같이 총 19개의 성분을 검출 및 분석하였다. As a result of UPLC-QTOF-MS/MS analysis, a total of 19 components were detected and analyzed as shown in FIG. 2 .
가노데릭산 eta (GAeta; RT = 15.179 분, C30H44O8, m/z 532), 부틸 가노데릭산 A (BGAA; RT = 16.183 분, C34H52O7, m/z 572), 가노데릭산 C2 (GAC2; RT = 17.228 분, C30H46O7, m/z 517) 및 가노데릭산 A (GAA; RT = 28.159, C30H44O7, m/z 515)의 경우 유사한 쪼개짐 패턴을 확인하였다. 기존에 이 성분들의 경우 D링의 특징적인 쪼개짐이 보고되어 있는 바 이를 정성 분석하였다.Ganoderic acid eta (GAeta; RT = 15.179 min, C30H44O8, m/z 532), Butyl Ganoderic acid A (BGAA; RT = 16.183 min, C34H52O7, m/z 572), Ganoderic acid C2 (GAC2; RT = A similar cleavage pattern was confirmed for 17.228 min, C30H46O7, m/z 517) and Ganoderic acid A (GAA; RT = 28.159, C30H44O7, m/z 515). In the case of these components, the characteristic cleavage of the D-ring was previously reported, and this was analyzed qualitatively.
가노데릭산 G (GAG; RT = 20.556 분, C30H44O8, m/z 532), 가노드레닉산 B (GDAB; RT = 20.965 분, C30H42O7, m/z 514), 가노데릭산 B (GAB; RT = 22.753 분, C30H44O7, m/z 516), 가노드레닉산 K (GDAK; RT = 24.227 분, C32H44O9, m/z 572), 가노데릭산 K (GAK; RT = 25.604 분, C32H46O9, m/z 574), 가노드레닉산 D (GDAD; RT = 34.453 분, C30H44O7, m/z 512) 및 가노데릭산 D (GAD; RT = 38.331 분, C30H42O7, m/z 514)의 경우 탄소번호 7번과 15번에 하이드록기가 붙은 형태로 C링과 D링의 특징적인 쪼개짐 패턴을 보이는 것으로 보고되어 있다.Ganoderic acid G (GAG; RT = 20.556 min, C30H44O8, m/z 532), Ganoderic acid B (GDAB; RT = 20.965 min, C30H42O7, m/z 514), Ganoderic acid B (GAB; RT = 22.753) min, C30H44O7, m/z 516), Ganoderic acid K (GDAK; RT = 24.227 min, C32H44O9, m/z 572), Ganoderic acid K (GAK; RT = 25.604 min, C32H46O9, m/z 574), For ganodernic acid D (GDAD; RT = 34.453 min, C30H44O7, m/z 512) and ganoderic acid D (GAD; RT = 38.331 min, C30H42O7, m/z 514), the hydrogens at
그 외에 가노데릭산 H (GAH; RT = 28.731 분, C32H44O9, m/z)와 유사한 골격을 가지는 가노데릭산 C6 (가노데릭산 H의 탄소 12번이 탈아세틸화된 형태), 및 루시데닉산 D (가노데릭산 H의 탄소 3번이 산화되어 알코올기가 붙은 형태)는 D링의 특징적인 쪼재김 패턴과 가노데릭산 H의 쪼재김 패턴을 참고하여 분석하였다.In addition, ganoderic acid H (GAH; RT = 28.731 min, C32H44O9, m/z) and ganoderic acid C6 (deacetylated form of
(3) 반정량 분석(3) semi-quantitative analysis
위의 정성 분석 결과를 토대로 Agilent MassHunter Workstation 프로그램을 이용하여 반정량 분석을 실시하였다. 정량분석 결과는 하기 표 2-4 및 도 3과 같았다.Based on the above qualitative analysis results, semi-quantitative analysis was performed using the Agilent MassHunter Workstation program. Quantitative analysis results were shown in Table 2-4 and FIG. 3 below.
±123,225687,985
±123,225
±109,382441,578
±109,382
±49,710486,832
±49,710
±143,358754,764
±143,358
±143,358635,989
±143,358
±1,127,0363,085,383
±1,127,036
±354,7191,908,072
±354,719
±629,6611,838,681
±629,661
±293,5212,055,515
±293,521
±127,6581,618,223
±127,658
±989,6771,928,533
±989,677
±234,9131,234,696
±234,913
±454,2881,134,606
±454,288
±382,6961,958,492
±382,696
±69,7951,443,624
±69,795
±6,645,4079,644,429
±6,645,407
±1,058,3763,285,273
±1,058,376
±1,503,9942,441,869
±1,503,994
±386,1654,006,304
±386,165
±63,6703,161,750
±63,670
±1,154,6553,867,549
±1,154,655
±333,7582,990,578
±333,758
±459,9262,688,483
±459,926
±316,6434,027,345
±316,643
±170,2643,242,556
±170,264
±606,5581,276,931
±606,558
±855,4121,444,309
±855,412
±746,7622,228,511
±746,762
±861,3941,834,914
±861,394
±613,2086,321,831
±613,208
±771,4395,884,623
±771,439
±1,170,5759,117,802
±1,170,575
±712,0286,664,291
±712,028
±560,8496,283,350
±560,849
±342,2695,828,423
±342,269
±1,590,8517,870,545
±1,590,851
±1,170,6145,187,842
±1,170,614
±744,8434,507,415
±744,843
±203,6563,745,784
±203,656
±782,1136,249,796
±782,113
±66,3524,916,080
±66,352
±173,409412,156
±173,409
±173,409225,836
±173,409
±82,178437,149
±82,178
±78,717292,770
±78,717
±1,283,8621,486,360
±1,283,862
±505,2111,126,862
±505,211
±505,211771,711
±505,211
±218,8491,361,551
±218,849
±121.041891,869
±121.041
±17,348,85918,509,502
±17,348,859
±2,244,61919,668,856
±2,244,619
±2,244,61917,761,856
±2,244,619
±2,996,65625,304,038
±2,996,656
±5,653,63719,011,430
±5,653,637
±499,139816,581
±499,139
±142,435455,234
±142,435
±45,054221,516
±45,054
±217,656582,234
±217,656
±126,799492,103
±126,799
±105,905414,670
±105,905
±257,206236,548
±257,206
±257,206217,720
±257,206
±246,776419,209
±246,776
±72,220328,699
±72,220
±1,409,6041,653,494
±1,409,604
±53,344270,295
±53,344
±131,570216,907
±131,570
±274,630707,365
±274,630
±221,051271,856
±221,051
±121,733542,856
±121,733
±138,904562,522
±138,904
±26,843532,654
±26,843
±23,875664,083
±23,875
±146,692532,835
±146,692
±247,8002001,953
±247,800
±448,3722,121,054
±448,372
±143,4361,949,150
±143,436
±78,3951,904,119
±78,395
±149,2271,901,536
±149,227
±230,5291,071,238
±230,529
±223,6721,495,767
±223,672
±216,3081,132,470
±216,308
±60,5201,622,025
±60,520
±111,5991,652,953
±111,599
±200,2902,382,995
±200,290
±527,6063,437,828
±527,606
±200,2003,120,484
±200,200
±98,3573,721,425
±98,357
±789,9974,026,071
±789,997
±659,2313,266,650
±659,231
±519,1773,672,913
±519,177
±330,4833,408,274
±330,483
±123,5053,782,335
±123,505
±358,4312,954,402
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±281,1431,589,884
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±96,0601,502,420
±96,060
±778,4512,307,602
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±1,070,7626,336,495
±1,070,762
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±1,792,8846,487,146
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578,7094,268,714
578,709
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±29,822
각 추출 조건에 따라서 가노데릭산 성분들의 함량과 비율이 다르게 나타나는 것을 확인하였다. It was confirmed that the content and ratio of ganoderic acid components were different according to each extraction condition.
특히 40℃와 55℃ 추출물의 경우 시간에 따른 가노데릭산 성분들의 함량이 다른 것으로 나타났으며, 12시간 추출물에서 GAG, GAB, GDAK, GAK 및 GAH 의 함량이 가장 높게 나타났다. 하지만 24시간의 장기간 추출 시 성분들의 함량 감소가 확인되었다. 이에 따라 가노데릭산을 추출하는데 적합한 추출 시간 또한 다른 것을 알 수 있었다.In particular, in the case of 40℃ and 55℃ extracts, the contents of ganoderic acid components were different according to time, and the contents of GAG, GAB, GDAK, GAK and GAH were the highest in the 12-hour extract. However, it was confirmed that the content of the components decreased during extraction for 24 hours. Accordingly, it was found that the extraction time suitable for extracting ganoderic acid was also different.
실험예 2: 세포 독성 효과 확인Experimental Example 2: Confirmation of cytotoxic effect
실시예 1에서 제조한 가노데릭산 추출물에 대하여 세포 배양을 이용한 세포 증식 효과를 측정하였다. For the ganoderic acid extract prepared in Example 1, the cell proliferation effect using cell culture was measured.
세포 독성 실험은 문헌(Mosmann T. et al, J. Immunol. Meth., 65 55-63(1983)에 기재된 방법에 따라 실시하였다. 세포 증식 및 독성 시험에 사용한 배지는 10% FBS(fetal bovine serum, Gibco)를 포함하는 DMEM(Dulbeco"s Modified Eagle"s Medium/high glucose, Gibco)에 성인 피부 세포주(KCLB, HaCaT; Keratinocyte cell)를 96-웰 플레이트에 1 x 102 세포/㎖의 수로 접종하여 사용하였다. 접종 24시간 후 FBS가 첨가되지 않은 배지로 교체하고 가노데릭산 추출물을 농도별 (3, 10, 30 ug/㎖)로 첨가한 후 24시간 동안 37℃ 5% CO2 배양기(incubator)에 배양하였으며, 대조군은 배양액만 넣어주었다. 배양이 끝난 후 DPBS를 통해 세포를 워싱하고 0.5 mg/㎖ 농도의 MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5- Diphenyltetrazolium Bromide, Dojindo Laboratories ZY903) 용액을 (1 ㎖/웰) 첨가한 후 37℃ CO2 배양기에서 2시간 후, 배지를 제거하고 DPBS로 다시 한 번 워싱했다. 여기에 DMSO(dimethylsulfoxide, Sigma D 4540)를 100 ㎕씩 첨가한 뒤 550 nm에서 흡광도를 측정하여 하기 수학식 1의 계산식에 따라 세포 독성율을 계산하였다. Cytotoxicity experiments were performed according to the method described in Mosmann T. et al, J. Immunol. Meth., 65 55-63 (1983). The medium used for cell proliferation and toxicity testing was 10% fetal bovine serum (FBS). , Gibco) containing adult skin cell lines (KCLB, HaCaT; Keratinocyte cells) in DMEM (Dulbeco's Modified Eagle's Medium/high glucose, Gibco) in a 96-well plate at a number of 1 x 10 2 cells/ml. After 24 hours of inoculation, the medium was replaced with FBS-free medium, and ganoderic acid extract was added at each concentration (3, 10, 30 ug/ml), and then 37° C. 5% CO 2 incubator for 24 hours. ), and the control group added only the culture solution After the culture was completed, the cells were washed with DPBS and MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium at a concentration of 0.5 mg/ml Bromide, Dojindo Laboratories ZY903) solution (1 ml/well) was added, and after 2 hours in a 37° C. CO 2 incubator, the medium was removed and washed again with DPBS. After adding 100 μl of DMSO (dimethylsulfoxide, Sigma D 4540) to this, absorbance was measured at 550 nm to calculate the cytotoxicity rate according to the formula in
수학식 1
실험결과는 도 4에 나타내었다.The experimental results are shown in FIG. 4 .
도 4의 결과에서 알 수 있는 바와 같이 본 발명의 가노데릭산 추출물을 함유하는 추출물의 경우 농도가 높아질수록 세포 생존률이 낮아짐을 알 수 있었고, 30 ug/㎖의 농도에서 독성을 나타냄을 알 수 있었다. 또한, 추출 조건에 따라 세포 독성이 다르게 나타남을 알 수 있었다.As can be seen from the results of Figure 4, in the case of the extract containing the ganoderic acid extract of the present invention, it was found that the higher the concentration, the lower the cell viability, and it was found that the toxicity at a concentration of 30 ug / ml. . In addition, it was found that the cytotoxicity was different depending on the extraction conditions.
실험예 3: 유전자 발현 분석Experimental Example 3: Gene expression analysis
세포 증식 및 독성 시험에 사용한 배지는 10% FBS(fetal bovine serum, Gibco)를 포함하는 DMEM(Dulbeco"s Modified Eagle"s Medium/high glucose, Gibco)에 성인 피부 각질 세포주(KCLB, HeCaT)를 6-웰 플 레이트에 1X106 세포/㎖의 수로 접종하여 사용하였다. 10 ug/㎖의 샘플과 섞어 한 시간 배양한 후 50 ng/㎖ rhIL-17a와 10 ng/㎖ rhTNF-α을 넣고 24시간 동안 배양하여 피부 염증을 유도하였다. The medium used for cell proliferation and toxicity tests was adult skin keratinocytes (KCLB, HeCaT) in DMEM (Dulbeco's Modified Eagle's Medium/high glucose, Gibco) containing 10% fetal bovine serum (Gibco). - The well plate was inoculated at a number of 1X10 6 cells/ml and used. After mixing with a 10 ug/ml sample and incubating for 1 hour, 50 ng/ml rhIL-17a and 10 ng/ml rhTNF-α were added and incubated for 24 hours to induce skin inflammation.
RNeasy kit (Qiagen, MD, USA)을 이용해 RNA를 추출한 후, 1 ug의 RNA를 역전사 시스템 (cDNA with RevertAid First Strand cDNA Synthesis Kit; Thermo Fisher Scientific, MA, USA)에 이용해 cDNA를 합성하였다. 이후 상기 cDNA를 이용해 정량적 PCR을 실시해 각 유전자의 발현 수준을 측정하였다. 본 PCR에 쓰인 프라이머는 표 5에 나타내었다.After RNA was extracted using the RNeasy kit (Qiagen, MD, USA), cDNA was synthesized using 1 ug of RNA in a reverse transcription system (cDNA with RevertAid First Strand cDNA Synthesis Kit; Thermo Fisher Scientific, MA, USA). Thereafter, quantitative PCR was performed using the cDNA to measure the expression level of each gene. The primers used in this PCR are shown in Table 5.
SYBR green을 이용한 real-time PCR은 StepOne Real Time PCR System(Applied Biosystems, Foster City, CA, USA)을 이용하여 진행하였고, GAPDH를 하우스 키핑 유전자로 사용하여 comparative CT 방법으로 분석하였다. Real-time PCR using SYBR green was performed using the StepOne Real Time PCR System (Applied Biosystems, Foster City, CA, USA), and analyzed by comparative CT method using GAPDH as a housekeeping gene.
본 발명에 사용된 프라이머는 케모카인 (CCL5, CCL20)과 사이토카인 (IL-6, IL, IL-23/p19), S100 칼슘 결합 단백질들 (S100A7, S100A8, S100A9) 등의 아토피 피부염과 간질의 발생 기전에 영향을 미치는 것으로 보고된 단백질을 대상으로 하였다. The primers used in the present invention include chemokines (CCL5, CCL20), cytokines (IL-6, IL, IL-23/p19), and S100 calcium binding proteins (S100A7, S100A8, S100A9), such as atopic dermatitis and epilepsy Proteins reported to affect the mechanism were targeted.
실험결과는 도 5 및 도 6과 같았다. 도 5 및 도 6의 결과에서 알 수 있는 바와 같이, 인터루킨-6 (IL-6), S100A7, 등의 발현 억제 활성이 본 발명의 가노데릭산 추출물의 경우 다른 조건에서는 낮았으나 40℃에서 12시간과 55℃에서 1-24시간 추출물의 경우 mRNA 수준에서 우수한 항염증 활성을 확인할 수 있었다. Experimental results were as shown in FIGS. 5 and 6 . As can be seen from the results of Figures 5 and 6, the expression inhibitory activity of interleukin-6 (IL-6), S100A7, etc. was low in the case of the ganoderic acid extract of the present invention under other conditions, but at 40 °C for 12 hours. And in the case of the extract for 1-24 hours at 55°C, excellent anti-inflammatory activity was confirmed at the mRNA level.
40℃에서 12시간의 특정 조건에서 mRNA 수준에서 우수한 항염증 활성이 인터루킨-6 (IL-6), S100A7, 등의 발현 억제로 확인되었다. 55℃에서는 6시간, 12시간 조건에서 IL-6 발현 억제 활성이 1시간, 3시간, 6시간, 12시간, 24시간의 모든 조건에서 유의적인 S100A7 억제 활성을 보였다. 70℃ 조건의 경우 단시간 추출인 1시간, 3시간 조건에서 S100A7의 억제 활성이 확인되었다. 이에 따라 40℃ 12시간, 55℃ 12시간24시간, 70℃ 1시간의 우수한 활성을 보인 조건에서 추출된 추출물을 대상으로 다양한 유전자 발현 활성을 확인하였다. Excellent anti-inflammatory activity at the mRNA level under specific conditions for 12 hours at 40°C was confirmed by inhibition of expression of interleukin-6 (IL-6), S100A7, and the like. At 55°C, the IL-6 expression inhibitory activity under the conditions of 6 hours and 12 hours showed significant S100A7 inhibitory activity at all the conditions of 1 hour, 3 hours, 6 hours, 12 hours, and 24 hours. In the case of 70 ℃ condition, the inhibitory activity of S100A7 was confirmed under the short-time extraction conditions of 1 hour and 3 hours. Accordingly, various gene expression activities were confirmed for the extracts extracted under the conditions showing excellent activity at 40° C. for 12 hours, 55° C. for 12 hours and 24 hours, and at 70° C. for 1 hour.
피부 염증의 경우 활성화되는 경로가 복잡해 다양한 프라이머를 이용해 항염증 활성에 효과가 있는지를 확인하였다. 실험결과는 도 7과 같았다. 도 7에 나타난 바와 같이 40℃에서 12시간 추출한 영지버섯 추출물의 경우 높은 항염증 활성을 보였으며, 특히 CCL5, S100A8, S100A9의 경우 음성 대조군과 비교했을 때 그 활성이 우수함을 알 수 있었다.In the case of skin inflammation, the activation pathway is complex, so it was confirmed whether various primers were effective in anti-inflammatory activity. The experimental results were as shown in FIG. 7 . As shown in FIG. 7 , the reishi mushroom extract extracted at 40° C. for 12 hours showed high anti-inflammatory activity, and in particular, CCL5, S100A8, and S100A9 showed superior activity compared to the negative control group.
이상, 첨부된 도면을 참조하여 본 발명의 실시예를 설명하였지만, 본 발명이 속하는 기술분야에서 통상의 지식을 가진 자는 본 발명이 그 기술적 사상이나 필수적인 특징으로 변경하지 않고서 다른 구체적인 형태로 실시될 수 있다는 것을 이해할 수 있을 것이다. 그러므로 이상에서 기술한 실시예는 모든 면에서 예시적인 것이며 한정적이 아닌 것으로 이해해야만 한다.As mentioned above, although embodiments of the present invention have been described with reference to the accompanying drawings, those of ordinary skill in the art to which the present invention pertains can practice the present invention in other specific forms without changing its technical spirit or essential features. You will understand that there is Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive.
<110> Korea Institute of Science and Technology <120> Composition for treatment, prevention or reducing atopic dermatitis, psoriasis or dermatitis comprising ganoderic acid <130> DP-2020-0028 <160> 22 <170> KoPatentIn 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_IL-1b_F <400> 1 tgagctcgcc agtgaaatga 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_IL-1b_R <400> 2 agattcgtag ctggatgccg 20 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_IL-6_F <400> 3 ttcggtccag ttgccttctc 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_IL-6_R <400> 4 tcttctcctg ggggtactgg 20 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_IL-23p19_F <400> 5 acagaagctc tgcacactgg 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_IL-23p19_R <400> 6 gttgtccctg agtccttggg 20 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_IFN-y_F <400> 7 gggaagcgaa caaggagtca 20 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_IFN-y_R <400> 8 ctgggatgct cttcgacctc 20 <210> 9 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_TNF-a_F <400> 9 tcttctcgaa ccccgagtga 20 <210> 10 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_TNF-a_R <400> 10 gggtttgcta caacatgggc 20 <210> 11 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_CCL5_F <400> 11 cagtcgtctt tgtcacccga 20 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_CCL5_R <400> 12 tcttctctgg gttggcacac 20 <210> 13 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_CCL20_F <400> 13 tgtcagtgct gctactccac 20 <210> 14 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_CCL20_R <400> 14 ccgtgtgaag cccacaataa 20 <210> 15 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_S100A7_F <400> 15 gcacaaatta cctcgccgat 20 <210> 16 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_S100A7_R <400> 16 tgcttgtggt agtctgtggc 20 <210> 17 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_S100A8_F <400> 17 aaggggaatt tccatgccgt 20 <210> 18 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_S100A8_R <400> 18 aggacactcg gtctctagca 20 <210> 19 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_S100A9_F <400> 19 catgctgatg gcgaggctaa 20 <210> 20 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_S100A9_R <400> 20 ccctcgtgca tcttctcgtg 20 <210> 21 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_GAPDH_F <400> 21 gaaggtgaag gtcggagtc 19 <210> 22 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_GAPDH_R <400> 22 gaagatggtg atgggatttc 20 <110> Korea Institute of Science and Technology <120> Composition for treatment, prevention or reducing atopic dermatitis, psoriasis or dermatitis comprising ganoderic acid <130> DP-2020-0028 <160> 22 <170> KoPatentIn 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_IL-1b_F <400> 1 tgagctcgcc agtgaaatga 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_IL-1b_R <400> 2 agattcgtag ctggatgccg 20 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_IL-6_F <400> 3 ttcggtccag ttgccttctc 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_IL-6_R <400> 4 tcttctcctg ggggtactgg 20 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_IL-23p19_F <400> 5 acagaagctc tgcacactgg 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_IL-23p19_R <400> 6 gttgtccctg agtccttggg 20 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_IFN-y_F <400> 7 gggaagcgaa caaggagtca 20 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_IFN-y_R <400> 8 ctgggatgct cttcgacctc 20 <210> 9 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_TNF-a_F <400> 9 tcttctcgaa ccccgagtga 20 <210> 10 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_TNF-a_R <400> 10 gggtttgcta caacatgggc 20 <210> 11 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_CCL5_F <400> 11 cagtcgtctt tgtcacccga 20 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_CCL5_R <400> 12 tcttctctgg gttggcacac 20 <210> 13 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_CCL20_F <400> 13 tgtcagtgct gctactccac 20 <210> 14 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_CCL20_R <400> 14 ccgtgtgaag cccacaataa 20 <210> 15 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_S100A7_F <400> 15 gcacaaatta cctcgccgat 20 <210> 16 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_S100A7_R <400> 16 tgcttgtggt agtctgtggc 20 <210> 17 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_S100A8_F <400> 17 aaggggaatt tccatgccgt 20 <210> 18 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_S100A8_R <400> 18 aggacactcg gtctctagca 20 <210> 19 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_S100A9_F <400> 19 catgctgatg gcgaggctaa 20 <210> 20 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_S100A9_R <400> 20 ccctcgtgca tcttctcgtg 20 <210> 21 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_GAPDH_F <400> 21 gaaggtgaag gtcggagtc 19 <210> 22 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_H_Q_GAPDH_R <400> 22 gaagatggtg atgggatttc 20
Claims (15)
A composition for preventing, treating or alleviating skin inflammation comprising ganoderic acid or a derivative thereof as an active ingredient.
A composition for preventing, treating or alleviating atopy comprising ganoderic acid or a derivative thereof as an active ingredient.
A composition for preventing, treating or alleviating psoriasis comprising ganoderic acid or a derivative thereof as an active ingredient.
상기 가노데릭산 또는 이의 유도체는
가노데릭산 eta (GAeta), 부틸 가노데릭산 A (BGAA), 가노데릭산 C2 (GAC2), 가노데릭산 A (GAA), 가노데릭산 G (GAG), 가노드레닉산 B (GDAB), 가노데릭산 B (GAB), 가노드레닉산 K (GDAK), 가노데릭산 K (GAK), 가노드레닉산 D (GDAD), 가노데릭산 D (GAD), 가노데릭산 H (GAH), 가노데릭산 C6, 아세틸 가노데릭산 F 및 루시데닉산 D 중에서 선택된 어느 하나 이상인, 조성물.
4. The method according to any one of claims 1 to 3,
The ganoderic acid or its derivative is
Ganoderic acid eta (GAeta), Butyl Ganoderic acid A (BGAA), Ganoderic acid C2 (GAC2), Ganoderic acid A (GAA), Ganoderic acid G (GAG), Ganoderic acid B (GDAB), Gano Deric Acid B (GAB), Ganoderic Acid K (GDAK), Ganoderic Acid K (GAK), Ganoderic Acid D (GDAD), Ganoderic Acid D (GAD), Ganoderic Acid H (GAH), Ganoderic Acid C6, acetyl ganoderic acid F, and at least one selected from lucidenic acid D, the composition.
상기 가노데릭산 또는 이의 유도체는 영지버섯(ganoderma lucidum)의 추출물로부터 분리 또는 수득된 것인, 조성물.
4. The method according to any one of claims 1 to 3,
The ganodermic acid or its derivative is isolated or obtained from an extract of reishi mushroom ( ganoderma lucidum ), the composition.
상기 영지버섯의 추출물은 에탄올 추출물인, 조성물.
6. The method of claim 5,
The extract of the reishi mushroom is an ethanol extract, composition.
상기 영지버섯의 추출물은 30 내지 80℃의 온도에서, 2 내지 40 시간 동안 추출된 것인, 조성물.
6. The method of claim 5,
The extract of the reishi mushroom will be extracted for 2 to 40 hours at a temperature of 30 to 80 ℃, the composition.
상기 유효성분은 케모카인, 사이토카인 및 S100 칼슘 결합 단백질 중 어느 하나 이상을 억제하는, 조성물.
4. The method according to any one of claims 1 to 3,
The active ingredient inhibits any one or more of chemokines, cytokines and S100 calcium binding protein, the composition.
상기 케모카인은 CCL5 및 CCL20 중 어느 하나 이상이며,
상기 사이토카인은 IL-6 및 IL-23/p19 중 어느 하나 이상이며,
상기 S100 칼슘 결합 단백질은 S100A7, S100A8 및 S100A9 중 어느 하나 이상인, 조성물.
9. The method of claim 8,
The chemokine is any one or more of CCL5 and CCL20,
The cytokine is any one or more of IL-6 and IL-23/p19,
The S100 calcium binding protein is any one or more of S100A7, S100A8 and S100A9, the composition.
상기 조성물은 약학 조성물인, 조성물.
4. The method according to any one of claims 1 to 3,
The composition is a pharmaceutical composition.
상기 조성물은 화장료 조성물인, 조성물.
4. The method according to any one of claims 1 to 3,
The composition is a cosmetic composition, composition.
상기 조성물은 건강기능 식품 조성물인, 조성물.
4. The method according to any one of claims 1 to 3,
The composition is a health functional food composition, composition.
영지버섯(ganoderma lucidum)을 30 내지 80℃의 온도에서, 2 내지 40 시간 동안 에탄올 용매를 이용하여 추출하는 단계를 포함하는, 제조 방법.
In the method for producing ganoderic acid or a derivative thereof for preventing, treating or alleviating skin inflammation, atopic dermatitis or psoriasis,
Reishi mushroom ( ganoderma lucidum ) at a temperature of 30 to 80 ℃, comprising the step of extracting using an ethanol solvent for 2 to 40 hours, a manufacturing method.
상기 추출하는 단계는 초음파 추출인, 제조 방법.
14. The method of claim 13,
The extracting step is ultrasonic extraction, the manufacturing method.
상기 방법은 추출 후 수득된 조추출물을 재용해하는 단계를 더 포함하는, 제조 방법.
14. The method of claim 13,
The method further comprises the step of re-dissolving the crude extract obtained after extraction.
Priority Applications (3)
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KR1020200041577A KR20210123853A (en) | 2020-04-06 | 2020-04-06 | Composition for treatment, prevention or reducing atopic dermatitis, psoriasis or dermatitis comprising ganoderic acid |
CN202110202078.1A CN113491701A (en) | 2020-04-06 | 2021-02-23 | Composition containing ganoderic acid for treating, preventing or reducing atopic disease, psoriasis or skin inflammation, and its preparation method |
KR1020220098153A KR102608571B1 (en) | 2020-04-06 | 2022-08-05 | Composition for treatment, prevention or reducing atopic dermatitis, psoriasis or dermatitis comprising ganoderic acid |
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KR1020200041577A KR20210123853A (en) | 2020-04-06 | 2020-04-06 | Composition for treatment, prevention or reducing atopic dermatitis, psoriasis or dermatitis comprising ganoderic acid |
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KR1020220098153A KR102608571B1 (en) | 2020-04-06 | 2022-08-05 | Composition for treatment, prevention or reducing atopic dermatitis, psoriasis or dermatitis comprising ganoderic acid |
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KR20090018886A (en) | 2006-03-23 | 2009-02-24 | 허발사이언스 싱가포르 피티이 리미티드 | Extracts and methods comprising ganoderma species |
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JPH11180889A (en) * | 1997-12-15 | 1999-07-06 | Buhei Kono | Agent for treating dermatitis containing extracted ingredient from ganoderma lucidum karst. |
TWI439273B (en) * | 2012-12-19 | 2014-06-01 | Microbiological Res Foundation R O C | Uses of ganoderic acids for preventing myocardial injury or damage |
JP2016013981A (en) * | 2014-07-01 | 2016-01-28 | 国立大学法人九州大学 | Tubulin polymerization activity regulator, food and agent comprising the same, and method of producing tubulin polymerization activity regulator |
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KR20090018886A (en) | 2006-03-23 | 2009-02-24 | 허발사이언스 싱가포르 피티이 리미티드 | Extracts and methods comprising ganoderma species |
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