KR102305964B1 - A composition for immune enhancement comprising narrow-leaf erecta fig extract - Google Patents
A composition for immune enhancement comprising narrow-leaf erecta fig extract Download PDFInfo
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- KR102305964B1 KR102305964B1 KR1020190160388A KR20190160388A KR102305964B1 KR 102305964 B1 KR102305964 B1 KR 102305964B1 KR 1020190160388 A KR1020190160388 A KR 1020190160388A KR 20190160388 A KR20190160388 A KR 20190160388A KR 102305964 B1 KR102305964 B1 KR 102305964B1
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- extract
- roasted
- chrysanthemum
- narrowleaf
- narrow
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Abstract
본 발명은 좁은잎천선과 추출물을 유효성분으로 포함하는 면역 증진용 조성물에 관한 것이다.The present invention relates to a composition for enhancing immunity, comprising an extract of the Narrowleaf chrysanthemum extract as an active ingredient.
Description
본 발명은 좁은잎천선과 추출물을 유효성분으로 포함하는 면역 증진용 조성물에 관한 것으로, 의약품 및 식품 등 다양한 용도로서 활용 가능하다.The present invention relates to a composition for enhancing immunity, comprising an extract of Narrowleaf chrysanthemum as an active ingredient, and can be used for various purposes such as pharmaceuticals and food.
모든 생물체는 생명을 영위하기 위해 끊임없이 많은 영양소를 필요로 하며, 이러한 영양소는 식품의 형태로 항상 외부로부터 공급되거나 일부는 체내에서 합성된다.All living things constantly need a lot of nutrients to sustain life, and these nutrients are always supplied from the outside in the form of food or some are synthesized in the body.
음식물 속에는 인간의 생명현상을 유지하는데 필요한 물질들이 포함되어 있으며, 인간은 음식물을 섭취하여 소화기관에서 소화시키고 흡수하여 체내의 조직세포에 공급함으로써 생명을 유지하고 있다.Food contains substances necessary for maintaining human life, and humans sustain life by ingesting food, digesting it in the digestive system, absorbing it, and supplying it to the tissue cells of the body.
따라서 좋은 영양은 건강한 신체를 유지시켜주며, 건강한 신체는 건강한 정신을 보장한다.Therefore, good nutrition maintains a healthy body, and a healthy body guarantees a healthy mind.
영양은 직접, 간접으로 질병의 원인이 되기도 하며, 질병의 예방 및 치료에도 관여하는 것으로 알려져 있는데, 인체에 필요한 영양소의 균형섭취 제대로 이루어지지 않아 생기는 영양분의 부족도 문제이고, 과잉섭취 또한 심각한 문제이며, 이로 인해 각종 질병이 유발될 수 있다.Nutrition is known to cause diseases directly or indirectly, and is also known to be involved in the prevention and treatment of diseases. , which can lead to various diseases.
최근 생활수준이 높아지고 식생활이 서구화되면서 영양과잉, 운동부족 등으로 인한 비만, 당뇨, 고혈압 및 심장병 등과 같은 성인병 발병률이 증가하고 있다. Recently, as the standard of living increases and the diet becomes westernized, the incidence of adult diseases such as obesity, diabetes, high blood pressure, and heart disease is increasing due to overnutrition and lack of exercise.
또한 평균수명 연장으로 인하여 인구가 점차 노령화되면서 건강에 대한 관심이 점차 높아지고 있다.In addition, as the population gradually ages due to the extension of life expectancy, interest in health is gradually increasing.
건강증진(health promotion)은 질병, 특히 만성 퇴행성질환이나 감염증, 계속되는 스트레스에 대한 저항력의 증가, 일상의 활동력의 증대를 꾀하는 것으로 이를 위해 영양의 개선, 면역기능 강화, 적절한 운동과 휴식, 정신적 활동의 지속 등 생활양식 전반에 미치는 관리가 요구된다. Health promotion aims to increase resistance to diseases, particularly chronic degenerative diseases or infectious diseases, and continuous stress, and to increase daily activity. Management that affects the overall lifestyle, such as sustainability, is required.
면역은 체내에 존재하는 자기방어체계로서 인체가 외부에서 침입하는 각종 물질이나 생명체를 자기 자신에 대한 이물질로 인식하여 제거하고 대사시키는 과정이다.Immunity is a self-defense system that exists in the body, and it is a process in which the human body recognizes and removes and metabolizes various substances or living things that invade from the outside as foreign substances against itself.
외부 자극에 의한 손상이나 병원 미생물의 침입으로부터 자신을 방어하기도 하지만 염증반응 등과 같이 자기 자신의 조직에 손상을 줄 수도 있다.It protects itself from damage caused by external stimuli or invasion of pathogenic microorganisms, but it can also damage its own tissues, such as an inflammatory response.
면역기능의 변화를 조절하여 정상으로 회복시키거나 변화의 폭을 줄여 주는 작용으로 면역기능 억제나 면역기능 증강으로 구분된다.It modulates changes in immune function to restore normalcy or reduces the range of changes, and is classified as suppression of immune function or enhancement of immune function.
면역 기능에 영향을 미치는 요인에는 유전적, 환경적 요인, 나이, 성별, 심리적 스트레스, 영양상태, 식이습관, 질병 등이 있다.Factors that affect immune function include genetic and environmental factors, age, gender, psychological stress, nutritional status, dietary habits, and disease.
면역 기능의 조절 기능을 갖는 건강기능식품은 현재까지, 저하된 면역기능을 증진 시키는 기능성과 과도한 면역기능을 조절하는 기능성, 자기 성분에 대한 면역반응을 조절하는 자가면역기능 조절 기능성으로 구분된다.So far, health functional foods having a function of regulating immune function are divided into a function to enhance a reduced immune function, a function to regulate an excessive immune function, and a function to regulate an autoimmune function to regulate an immune response to self-components.
면역기능이 저하되면 과민반응을 일으키는 경우가 있는데 이러한 과민반응은 천식, 계절성 또는 동년성 비염, 알러지성 부비강염, 결막염, 식품과 약품에 의한 알러지, 아토피성 피부염, 두드러기, 벌침에 의한 알러지 등과 같이 다양하게 나타난다.When the immune function is lowered, hypersensitivity reactions may occur. Such hypersensitivity reactions are diverse, such as asthma, seasonal or same-year rhinitis, allergic sinusitis, conjunctivitis, food and drug allergy, atopic dermatitis, urticaria, and allergy to bee stings. appear to be
여러 원인으로 인한 면역기능 장애를 극복하기 위하여, 다양한 면역 증강제가 사용되고 있으나, 부작용 및 생체 안전성의 문제가 있으므로 장기 복용이 가능하고 예방용으로도 적합한 면역 증진 조성물이 요구되는 실정이다.In order to overcome immune dysfunction due to various causes, various immune enhancing agents are used, but there is a need for an immune enhancing composition suitable for long-term use and prevention because there are side effects and biosafety problems.
(특허문헌 1) 대한민국등록특허 제10-2011684호
(특허문헌 2) 대한민국등록특허 제10-1874465호(Patent Document 1) Republic of Korea Patent No. 10-2011684
(Patent Document 2) Republic of Korea Patent No. 10-1874465
본 발명은 전술한 종래 기술의 문제점을 해결하기 위한 것으로, 본 발명의 목적은 생체 안전성이 우수한 천연 식물 유래의 기능성 식의약품을 제공하는 것이다.The present invention is to solve the problems of the prior art described above, and an object of the present invention is to provide a functional food product derived from a natural plant having excellent biosafety.
본 발명의 일 측면에 따르면, 좁은잎천선과 추출물을 유효성분으로 포함하는 면역 증진용 조성물이 제공된다.According to one aspect of the present invention, there is provided a composition for enhancing immunity comprising the extract of chrysanthemum chrysanthemum as an active ingredient.
일 실시예에 있어서, 상기 추출물은 물, 알코올, 에틸아세테이트, 아세톤, 핵산, 디클로로메탄 또는 이들의 혼합 용매로 추출될 수 있다.In one embodiment, the extract may be extracted with water, alcohol, ethyl acetate, acetone, nucleic acid, dichloromethane, or a mixed solvent thereof.
본 발명의 다른 측면에 따르면, 로스팅(roasting) 좁은잎천선과 추출물을 유효성분으로 포함하는 면역 증진용 조성물이 제공된다.According to another aspect of the present invention, there is provided a composition for enhancing immunity comprising the extract of Roasting Narrowleaf chrysanthemum as an active ingredient.
일 실시예에 있어서, 상기 좁은잎천선과는 150 내지 300℃에서 로스팅할 수 있다.In one embodiment, the narrow leaf cheonseonwa may be roasted at 150 to 300 °C.
일 실시예에 있어서, 상기 좁은잎천선과는 250℃에서 150℃까지 온도를 조절하면서 로스팅할 수 있다.In one embodiment, the narrow leaf cheonseonwa may be roasted while controlling the temperature from 250°C to 150°C.
일 실시예에 있어서, 상기 추출물은 열수 추출(hot water extraction)될 수 있다.In one embodiment, the extract may be subjected to hot water extraction.
본 발명의 다른 측면에 따르면, 좁은잎천선과 열수 추출물을 유효성분으로 포함하는 면역 증진용 건강기능식품이 제공된다.According to another aspect of the present invention, there is provided a health functional food for enhancing immunity comprising the extract of Narrowleaf chrysanthemum and hot water as active ingredients.
본 발명의 일 측면에 따른 조성물은 천연 추출물을 유효성분으로 포함하여 생체 안전성이 우수할 뿐만 아니라 면역 증진 활성이 우수하므로 건강기능식품뿐만 아니라 면역 관련 질환의 치료 용도로서 유용하게 활용할 수 있다.Since the composition according to one aspect of the present invention contains a natural extract as an active ingredient, it has excellent biosafety as well as excellent immune enhancing activity, so it can be usefully used as a health functional food as well as for the treatment of immune-related diseases.
본 발명의 효과는 상기한 효과로 한정되는 것은 아니며, 본 발명의 상세한 설명 또는 청구범위에 기재된 발명의 구성으로부터 추론 가능한 모든 효과를 포함하는 것으로 이해되어야 한다.The effects of the present invention are not limited to the above effects, but it should be understood to include all effects that can be inferred from the configuration of the invention described in the detailed description or claims of the present invention.
도 1은 본 발명의 일 실시예에 따른 좁은잎천선과 추출물의 세포 독성을 평가한 결과이다. (A)는 좁은잎천선과 열수 추출물, (B)는 좁은잎천선과 에탄올 추출물, (C)는 250℃에서 로스팅된 좁은잎천선과 열수 추출물 (D)는 250℃에서 로스팅된 좁은잎천선과 에탄올 추출물, (E)는 200℃에서 로스팅된 좁은잎천선과 열수 추출물 (F)는 200℃에서 로스팅된 좁은잎천선과 에탄올 추출물, (G)는 단계적으로 로스팅한 좁은잎천선과 열수 추출물, (H)는 단계적으로 로스팅한 좁은잎천선과 에탄올 추출물이다.
도 2는 본 발명의 일 실시예에 따른 좁은잎천선과 추출물의 면역 증진 활성을 평가(NO assay)한 결과이다. (A)는 좁은잎천선과 열수 추출물, (B)는 좁은잎천선과 에탄올 추출물, (C)는 250℃에서 로스팅된 좁은잎천선과 열수 추출물 (D)는 250℃에서 로스팅된 좁은잎천선과 에탄올 추출물, (E)는 200℃에서 로스팅된 좁은잎천선과 열수 추출물 (F)는 200℃에서 로스팅된 좁은잎천선과 에탄올 추출물, (G)는 단계적으로 로스팅한 좁은잎천선과 열수 추출물, (H)는 단계적으로 로스팅한 좁은잎천선과 에탄올 추출물이다.
도 3은 본 발명의 일 실시예에 따른 좁은잎천선과 추출물의 면역 증진 활성(TNF-a 생성량)을 평가한 결과이다. (A)는 좁은잎천선과 열수 추출물, (B)는 좁은잎천선과 에탄올 추출물, (C)는 250℃에서 로스팅된 좁은잎천선과 열수 추출물 (D)는 250℃에서 로스팅된 좁은잎천선과 에탄올 추출물, (E)는 200℃에서 로스팅된 좁은잎천선과 열수 추출물 (F)는 200℃에서 로스팅된 좁은잎천선과 에탄올 추출물, (G)는 단계적으로 로스팅한 좁은잎천선과 열수 추출물, (H)는 단계적으로 로스팅한 좁은잎천선과 에탄올 추출물이다.1 is a result of evaluating the cytotoxicity of the extract according to an embodiment of the present invention. (A) is Narrow-leaved C. spp. and hot water extract, (B) is Narrow-leaved C. C. and ethanol extract, (C) is Roasted Narrow-leaved C. C. and hot water extract (D) Roasted at 250°C Ethanol extract, (E) Narrowleaf chrysanthemum roasted at 200℃, and hot water extract (F) is ethanolic extract roasted at 200℃, (G) Narrowleaf chinensis and hot water extract roasted in stages H) is a stepwise roasted Narrowleaf chrysanthemum extract and ethanol extract.
2 is a result of evaluation (NO assay) of the immune enhancing activity of the extract of Narrowleaf chrysanthemum according to an embodiment of the present invention. (A) is Narrow-leaved C. spp. and hot water extract, (B) is Narrow-leaved C. C. and ethanol extract, (C) is Roasted Narrow-leaved C. C. and hot water extract (D) Roasted at 250°C Ethanol extract, (E) Narrowleaf chrysanthemum roasted at 200℃, and hot water extract (F) is ethanolic extract roasted at 200℃, (G) Narrowleaf chinensis and hot water extract roasted in stages H) is a stepwise roasted Narrowleaf chrysanthemum extract and ethanol extract.
Figure 3 is a result of evaluating the immune enhancing activity (TNF-a production amount) of the extract according to an embodiment of the present invention. (A) is Narrow-leaved C. spp. and hot water extract, (B) is Narrow-leaved C. C. and ethanol extract, (C) is Roasted Narrow-leaved C. C. and hot water extract (D) Roasted at 250°C Ethanol extract, (E) Narrowleaf chrysanthemum roasted at 200℃, and hot water extract (F) is ethanolic extract roasted at 200℃, (G) Narrowleaf chinensis and hot water extract roasted in stages H) is a stepwise roasted Narrowleaf chrysanthemum extract and ethanol extract.
본 명세서에서 사용되는 용어는 본 발명에서의 기능을 고려하면서 가능한 현재 널리 사용되는 일반적인 용어들을 선택하였으나, 이는 당 분야에 종사하는 기술자의 의도 또는 판례, 새로운 기술의 출현 등에 따라 달라질 수 있다. The terms used in this specification have been selected as currently widely used general terms as possible while considering the functions in the present invention, which may vary depending on the intention or precedent of a person skilled in the art, the emergence of new technology, and the like.
또한, 특정한 경우는 출원인이 임의로 선정한 용어도 있으며, 이 경우 해당되는 발명의 설명 부분에서 상세히 그 의미를 기재할 것이다. 따라서 본 발명에서 사용되는 용어는 단순한 용어의 명칭이 아닌, 그 용어가 가지는 의미와 본 발명의 전반에 걸친 내용을 토대로 정의되어야 한다.In addition, in a specific case, there is a term arbitrarily selected by the applicant, and in this case, the meaning will be described in detail in the description of the corresponding invention. Therefore, the term used in the present invention should be defined based on the meaning of the term and the overall content of the present invention, rather than the name of a simple term.
다르게 정의되지 않는 한, 기술적이거나 과학적인 용어를 포함해서 여기서 사용되는 모든 용어들은 본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자에 의해 일반적으로 이해되는 것과 동일한 의미를 가지고 있다. 일반적으로 사용되는 사전에 정의되어 있는 것과 같은 용어들은 관련 기술의 문맥상 가지는 의미와 일치하는 의미를 가지는 것으로 해석되어야 하며, 본 출원에서 명백하게 정의하지 않는 한, 이상적이거나 과도하게 형식적인 의미로 해석되지 않는다. Unless defined otherwise, all terms used herein, including technical and scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Terms such as those defined in a commonly used dictionary should be interpreted as having a meaning consistent with the meaning in the context of the related art, and should not be interpreted in an ideal or excessively formal meaning unless explicitly defined in the present application. does not
수치 범위는 상기 범위에 정의된 수치를 포함한다. 본 명세서에 걸쳐 주어진 모든 최대의 수치 제한은 낮은 수치 제한이 명확히 쓰여져 있는 것처럼 모든 더 낮은 수치 제한을 포함한다. 본 명세서에 걸쳐 주어진 모든 최소의 수치 제한은 더 높은 수치 제한이 명확히 쓰여져 있는 것처럼 모든 더 높은 수치 제한을 포함한다. 본 명세서에 걸쳐 주어진 모든 수치 제한은 더 좁은 수치 제한이 명확히 쓰여져 있는 것처럼, 더 넓은 수치 범위 내의 더 좋은 모든 수치 범위를 포함할 것이다.Numerical ranges are inclusive of the values defined in that range. Every maximum numerical limitation given throughout this specification includes all lower numerical limitations as if the lower numerical limitation were expressly written. Every minimum numerical limitation given throughout this specification includes all higher numerical limitations as if the higher numerical limitation were expressly written. Any numerical limitation given throughout this specification shall include all numerical ranges within the broader numerical range, as if the narrower numerical limitation were expressly written.
이하, 본 발명의 실시예를 상세히 기술하나, 하기 실시예에 의해 본 발명이 한정되지 아니함은 자명하다.Hereinafter, examples of the present invention will be described in detail, but it is obvious that the present invention is not limited by the following examples.
본 발명의 일 측면에 따르면, 좁은잎천선과 추출물을 유효성분으로 포함하는 면역 증진용 조성물이 제공된다.According to one aspect of the present invention, there is provided a composition for enhancing immunity comprising the extract of chrysanthemum chrysanthemum as an active ingredient.
상기 “좁은잎천선과(Ficus erecta var. sieboldii)”는 무화과과에 속하는 식물로서 가는잎천선과나무라고도 하며 우리나라에서도 제주도와 남해안 지방, 특히, 제주도에 주로 자라는 나무이다.The "narrow-leafed sieboldii " is a plant belonging to the fig family and is also referred to as a thin-leaved fenugreek.
상기 좁은잎천선과는 바닷가 산기슭에서 자라며 높이 2 내지 4m이고, 나무껍질은 잿빛이 섞인 흰색이며 어두운 갈색 피목이 있고 털이 없다. 잎은 어긋나고 천선과 보다 좁은 바소꼴이며 길이 10~20㎝이다. 끝부분이 뾰족하고 가장자리는 밋밋하거나 거친 톱니가 난다.The narrow-leaved chrysanthemum grows at the foot of a mountain by the sea and is 2 to 4 m in height, and the bark is grayish white with dark brown bark and no hairs. Leaves are alternate phyllotaxis, narrower lanceolate, and 10-20 cm long. The tip is sharp and the edge is flat or coarsely serrated.
상기 좁은잎천선과의 곁맥은 5 내지 6쌍이고, 잎자루는 길이가 1 내지 4cm이다. 상기 좁은잎천선과는 민간에서는 오래전부터 보중, 익기, 건비, 화습, 강근장골, 소종, 활혈, 해독의 효능으로 알려져 있다. 또한, 류마티스 관절염, 중기허약, 기혈쇠미, 사디산언, 근골분리, 타박상, 경폐, 산후유즙결핍 등의 치료에 이용되어 왔다.The side veins of the Narrow-leaved Cheonenaceae are 5 to 6 pairs, and the petiole has a length of 1 to 4 cm. The narrow-leaved cheonseonseonwae has long been known in the folklore for its effects of balancing, ripening, dryness, hwasup, strong iliac crest, small bell, hwalyeol, and detoxification. In addition, it has been used in the treatment of rheumatoid arthritis, mid-stage weakness, qi and blood loss, sadisaneon, musculoskeletal separation, bruises, transpulmonary, postpartum milk deficiency, and the like.
상기 “면역(immunity)”은 생물체 내에서 병원체와 종양 세포 등을 탐지한 후 무력화하여 질병으로부터 생명체를 보호하는 기작을 의미하는 것으로, 상기 “면역 증진(Immunostimulation)”은 암 및 염증 등과 같은 다양한 질환에 대한 신체 방어 기전을 보강하는 일련의 치료학적 또는 예방학적 전략을 모두 포함할 수 있다.The “immunity” refers to a mechanism to protect a living organism from disease by detecting and neutralizing pathogens and tumor cells in an organism, and the “immunostimulation” refers to various diseases such as cancer and inflammation. It can include both therapeutic and prophylactic strategies that reinforce the body's defense mechanisms against
상기 “유효성분으로 포함하는”은 투여하고자 하는 개체에 대한 치료적 또는 예방적 효과를 제공하기에 충분한 양을 포함하는 것을 의미하며, 상기 좁은잎천선과 추출물은 생체 안전성이 우수하므로 당업자가 양적 상한을 적절히 조정할 수 있다.The "comprising as an active ingredient" means to include an amount sufficient to provide a therapeutic or prophylactic effect to the subject to be administered, and the extract of N. chrysanthemum extract has excellent biosafety, so that those skilled in the art can limit the quantitative upper limit. can be adjusted appropriately.
상기 “추출물”은 용매와 추출 원료를 특정 조건 하에서 접촉시킴으로써 추출 원료에 함유된 유효성분이 전이된 용매를 지칭하는 것으로, 천연물로부터 원료에 함유된 성분을 분리해낸 물질이라면, 추출 방법이나 성분의 종류와 무관하게 모두 포함할 수 있다. 예컨대, 물이나 유기용매를 이용하여 천연물로부터 용매에 용해되는 성분을 추출한 것, 천연물의 특정 성분, 예컨대 오일과 같은 특정 성분만을 추출하여 얻어진 것 등을 모두 포함할 수 있다.The “extract” refers to a solvent in which the active ingredient contained in the extraction raw material is transferred by contacting the solvent and the extraction raw material under specific conditions. All can be included regardless. For example, extracting a component soluble in a solvent from a natural product using water or an organic solvent, a specific component of a natural product, for example, extracting only a specific component such as oil, etc. may be included.
상기 추출물은 좁은잎천선과를 건조 후 분말화하거나, 열수추출법, 에탄올 추출법 등 종래 알려진 다양한 추출법에 의해 수득할 수 있다.The extract can be obtained by a variety of conventionally known extraction methods, such as dry, powdered, or hot water extraction method, ethanol extraction method of the Narrow-leaf Cheonsonaceae.
상기 추출물은 물, 알코올, 에틸아세테이트, 아세톤, 핵산, 디클로로메탄 또는 이들의 혼합 용매로 추출될 수 있고, 바람직하게는 정제수를 이용하여 열수 추출될 수 있다.The extract may be extracted with water, alcohol, ethyl acetate, acetone, nucleic acid, dichloromethane or a mixed solvent thereof, preferably hot water extraction using purified water.
상기 원료에 포함된 유효성분은 용매의 극성에 따라 추출 비율이 상이해질 수 있으므로, 용매에 따른 생리 활성 물질의 선택성을 고려하여 달리할 수 있다.Since the extraction ratio of the active ingredient included in the raw material may be different depending on the polarity of the solvent, it may be different in consideration of the selectivity of the physiologically active material according to the solvent.
상기 추출물은 추출 원료를 물로 수세한 후 건조 및 분쇄하여, 원료 중량의 8 내지 12배에 달하는 부피의 용매로 약 1 내지 24시간 동안 환류 순환 추출, 가압 추출, 초음파 추출 등 통상적인 방법으로 추출 및 여과하여 제조할 수 있다.The extract is extracted by conventional methods such as reflux circulation extraction, pressure extraction, ultrasonic extraction, etc. for about 1 to 24 hours with a solvent in a volume equivalent to 8 to 12 times the weight of the raw material by washing the raw material for extraction with water and then drying and pulverizing it It can be prepared by filtration.
상기 추출물은 감압 증류 또는 동결 건조 등과 같은 추가적인 공정에 의해 분말 상태로 수득할 수 있다.The extract may be obtained in a powder state by an additional process such as distillation under reduced pressure or freeze-drying.
상기 추출물은 통상적인 정제 과정을 거친 추출물도 포함할 수 있다. 예컨대, 상기 추출물은 일정한 분자량 컷-오프 값을 갖는 한외 여과막을 이용한 분리, 다양한 크로마토그래피(크기, 전하, 소수성 또는 친화성에 따른 분리를 위해 제작된 것)에 의한 분리 등 추가적으로 실시된 다양한 정제 방법을 통해 얻어진 분획물을 포함할 수 있다.The extract may also include an extract that has undergone a conventional purification process. For example, the extract is separated using an ultrafiltration membrane having a constant molecular weight cut-off value, and various purification methods are additionally performed such as separation by various chromatography (prepared for separation according to size, charge, hydrophobicity or affinity). It may include a fraction obtained through
상기 조성물은 식품 조성물, 예컨대 면역 증진용 건강기능식품으로서 사용될 수 있다.The composition may be used as a food composition, such as a health functional food for enhancing immunity.
상기 건강기능식품은 건강기능식품에 관한 법률에 따른 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 제조 및 가공한 식품을 의미하며, 상기 기능성은 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건 용도에 유용한 효과를 얻을 목적으로 섭취하는 것을 의미할 수 있다.The health functional food means a food manufactured and processed using raw materials or ingredients useful for the human body according to the Health Functional Food Act, and the functionality is to control nutrients or physiologically affect the structure and function of the human body. It may refer to ingestion for the purpose of obtaining useful effects for health purposes, such as action.
상기 조성물은 통상의 식품 첨가물을 포함할 수 있으며, 상기 식품 첨가물은 다른 규정이 없는 한 식품의약품안전처에 승인된 식품 첨가물 공전의 총칙 및 일반시험법 등에 따라 해당 품목에 관한 규격 및 기준에 의하여 적합성 여부를 판단할 수 있다.The composition may include normal food additives, and unless otherwise specified, the food additives are approved by the Ministry of Food and Drug Safety according to the standards and standards for the relevant items according to the general rules and general test methods for food additives approved by the Ministry of Food and Drug Safety. can determine whether
상기 식품 첨가물 공전에 기재된 품목은 예컨대 케톤류, 글리신, 구연산칼륨, 니코틴산, 계피산 등의 화학적 합성물, 감색소, 감초추출물, 결정셀룰로오스, 고량색소, 구아검 등의 천연첨가물, L-글루타민산나트륨 제제, 면류첨가알칼리제, 보존료제제, 타르색소제제 등의 혼합제제류를 들 수 있다.The items described in the Food Additives Codex include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid, depigmentation, licorice extract, crystalline cellulose, high pigment, natural additives such as guar gum, sodium L-glutamate preparation, noodles Mixed formulations, such as an additive alkali agent, a preservative agent, and a tar dye agent, are mentioned.
상기 조성물은 면역 증진을 위한 식품 및 음료 등에 다양하게 이용될 수 있으며, 예컨대, 각종 식품류, 음료, 껌, 차, 비타민 복합제, 건강기능성 보조 식품, 식품 첨가제 등에 사용될 수 있다.The composition can be used in a variety of foods and beverages for immune enhancement, for example, can be used in various foods, beverages, gum, tea, vitamin complexes, health functional supplements, food additives, and the like.
또한, 상기 건강기능식품은 면역 증진을 목적으로 정제, 과립, 분말, 캅셀, 액상의 용액 및 환으로 이루어진 군에서 선택된 어느 하나의 제형으로 제조 및 가공될 수 있다.In addition, the health functional food may be manufactured and processed into any one formulation selected from the group consisting of tablets, granules, powders, capsules, liquid solutions and pills for the purpose of enhancing immunity.
구체적으로 상기 정제 형태의 건강기능식품은 좁은잎천선과 추출물, 부형제, 결합제, 붕해제 및 다른 첨가제와의 혼합물을 통상의 방법으로 과립화한 후, 활택제 등을 넣어 압축 성형하거나, 상기 혼합물을 직접 압축 성형하여 제조할 수 있다. 또한, 상기 정제 형태의 건강기능식품은 필요에 따라 교미제 등을 함유할 수 있으며, 필요에 따라 적당한 제피제로 제피할 수도 있다.Specifically, the health functional food in the form of a tablet is granulated by a conventional method by granulating a mixture of Narrowleaf chrysanthemum extract, excipients, binders, disintegrants and other additives, and then compression molding by adding a lubricant, or the mixture. It can be manufactured by direct compression molding. In addition, the health functional food in the form of tablets may contain a mating agent, etc., if necessary, and may be coated with a suitable skinning agent if necessary.
상기 캅셀 형태의 건강기능식품 중 경질캅셀제는 통상의 경질캅셀에 좁은잎천선과 추출물 및 부형제 등의 첨가제와의 혼합물 또는 그의 입상물 또는 제피한 입상물을 충진하여 제조할 수 있으며, 연질캅셀제는 좁은잎천선과 추출물 및 부형제 등의 첨가제와의 혼합물을 젤라틴 등 캅셀기제에 충진하여 제조할 수 있다. 상기 연질캅셀제는 필요에 따라 글리세린 또는 솔비톨 등의 가소제, 착색제, 보존제 등을 함유할 수 있다.Among the health functional foods in the form of capsules, hard capsules can be prepared by filling conventional hard capsules with a mixture of Narrowleaf chrysanthemum and additives such as extracts and excipients, or their granules or coated granules. It can be prepared by filling a capsule base such as gelatin with a mixture of leaf chrysanthemum extract and additives such as excipients. The soft capsules may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, if necessary.
상기 환 형태의 건강기능식품은 좁은잎천선과 추출물, 부형제, 결합제, 붕해제 등의 혼합물을 적당한 방법으로 성형하여 조제할 수 있으며, 필요에 따라 백당이나 다른 적당한 제피제로 제피를, 또는 전분, 탈크 또는 적당한 물질로 환의를 입힐 수도 있다.The health functional food in the form of a ring can be prepared by molding a mixture of Narrowleaf chrysanthemum extract, excipients, binders, disintegrants, etc. by an appropriate method. Alternatively, the gown may be dressed with a suitable material.
상기 과립형태의 건강기능식품은 좁은잎천선과 추출물, 부형제, 결합제, 붕해제 등의 혼합물을 적당한 방법으로 입상으로 제조할 수 있으며, 필요에 따라 착향제, 교미제 등을 함유할 수 있다. The health functional food in the form of granules can be prepared in a granular form by a suitable method of a mixture of Narrowleaf chrysanthemum extract, excipients, binders, disintegrants, and the like, and may contain flavoring agents, flavoring agents, etc. as necessary.
또한, 상기 부형제, 결합제, 붕해제, 활택제, 교미제, 착향제 등에 대한 용어 정의는 당업계에 공지된 문헌에 기재된 것으로 그 기능 등이 동일 내지 유사한 것들을 포함할 수 있다.In addition, the term definitions for the excipients, binders, disintegrants, lubricants, flavoring agents, and the like are described in documents known in the art and may include those having the same or similar functions.
본 발명의 다른 측면에 따르면, 좁은잎천선과 추출물을 유효성분으로 포함하는 면역 증진용 약학적 조성물이 제공된다.According to another aspect of the present invention, there is provided a pharmaceutical composition for enhancing immunity comprising the extract of chrysanthemum chrysanthemum extract as an active ingredient.
또한, 상기 조성물은 면역활성을 증진시킴으로써, 면역결핍, 면역저하 또는 면역계 손상으로 인한 질환의 예방, 개선 또는 치료에 유용하게 이용될 수 있다In addition, the composition can be usefully used for preventing, improving or treating diseases caused by immunodeficiency, immunosuppression, or immune system damage by enhancing immune activity.
상기 “예방”은 병리학적 현상의 발생 빈도 또는 정도를 감소시키는 모든 행위를 의미한다. 예방은 완전할 수 있으며 또는 부분적일 수도 있다. 이 경우에는 개체 내의 면역력 저하로 인해 발생되는 증상이 상기 조성물을 사용 하지 않은 경우와 비교하여 감소하는 현상을 의미할 수 있다.The "prevention" means any action that reduces the frequency or degree of occurrence of a pathological phenomenon. Prevention may be complete or partial. In this case, it may mean a phenomenon in which the symptoms caused by the decrease in immunity within the individual are reduced compared to the case where the composition is not used.
상기 “치료”는 치료하고자 하는 대상 또는 세포의 천연 과정을 변경시키기 위하여 임상적으로 개입하는 모든 행위를 의미하며, 임상 병리 상태가 진행되는 동안 또는 이를 예방하기 위하여 수행할 수 있다. 목적하는 치료 효과는 질병의 발생 또는 재발을 예방하거나, 증상을 완화시키거나, 질병에 따른 모든 직접 또는 간접적인 병리학적 결과를 저하시키거나, 전이를 예방하거나, 질병 진행 속도를 감소시키거나, 질병 상태를 경감 또는 일시적 완화시키거나, 예후를 개선시키는 것을 포함할 수 있다.The "treatment" refers to any action that clinically intervenes to change the natural process of a subject or cell to be treated, and may be performed while a clinical pathology is in progress or to prevent it. The desired therapeutic effect is to prevent the occurrence or recurrence of a disease, alleviate symptoms, reduce any direct or indirect pathological consequences of the disease, prevent metastasis, reduce the rate of disease progression, or reduce the disease progression rate. alleviating or temporarily ameliorating the condition, or improving the prognosis.
상기 조성물은 경구적 전달, 비경구적 전달의 형태로 투여될 수 있다. 상기 조성물은 전신 또는 국소 투여될 수 있으며, 상기 투여는 경구 투여 및 비경구 투여를 포함할 수 있다. 상기 조성물은 적절한 투여 형태를 제공하도록 적합한 양의 약학적으로 허용되는 비히클 또는 담체와 함께 제형화될 수 있다.The composition may be administered in the form of oral delivery or parenteral delivery. The composition may be administered systemically or locally, and the administration may include oral administration and parenteral administration. The compositions may be formulated with a suitable amount of a pharmaceutically acceptable vehicle or carrier to provide an appropriate dosage form.
상기 조성물은 약학 조성물의 제조에 사용되는 담체, 부형제 및 희석제를 더 포함할 수 있다. 상기 담체, 부형제 및 희석제로는 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 또는 광물유를 들 수 있으나, 이에 제한되는 것은 아니다.The composition may further include a carrier, excipient and diluent used in the preparation of the pharmaceutical composition. The carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil.
또한, 상기 조성물은 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제제화하여 사용할 수 있다.In addition, the composition can be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injection solutions.
경구 투여를 위한 고형제제는 정제, 환제, 산제, 과립제, 캡슐제 등이 사용될 수 있고, 상기 고형제제는 상기 좁은잎천선과 추출물과 이의 분획물들에 적어도 하나 이상의 부형제, 예컨대, 전분, 칼슘카보네이트, 수크로스, 락토오스, 또는 젤라틴 등을 혼합하여 조제할 수 있다. 또한, 상기 부형제 이외에 마그네슘 스티레이트, 탈크 같은 윤활제가 사용될 수 있다.A solid preparation for oral administration may include tablets, pills, powders, granules, capsules, etc., and the solid preparation may include at least one excipient, such as starch, calcium carbonate, It can be prepared by mixing sucrose, lactose, or gelatin. In addition to the above excipients, lubricants such as magnesium stearate and talc may be used.
경구 투여를 위한 액상 제제는 현탁제, 내용액제, 유제, 시럽제 등이 사용될 수 있고, 단순희석제인 물, 리퀴드 파라핀 외에 여러 가지 부형제, 예컨대 습윤제, 감미제, 방향제, 보존제 등이 사용될 수 있다.Liquid formulations for oral administration may include suspensions, solutions, emulsions, syrups, etc., and various excipients such as wetting agents, sweeteners, fragrances, preservatives, etc. may be used in addition to simple diluents such as water and liquid paraffin.
비경구 투여를 위한 제제는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제가 사용될 수 있다. 상기 비수성용제, 현탁제는 프로필렌글리콜(propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르가 사용될 수 있다. 상기 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로제라틴이 사용될 수 있다.Formulations for parenteral administration may be sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized formulations, and suppositories. The non-aqueous solvent and suspension may be propylene glycol, polyethylene glycol, vegetable oil such as olive oil, or injectable ester such as ethyl oleate. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin oil, and glycerogelatin may be used.
상기 약학적 조성물은 약제학적으로 유효한 양이 대상체에 투여될 수 있다. 상기 “약제학적으로 유효한 양”은 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분한 양을 의미하며, 유효 용량 수준은 환자의 질환의 종류, 중증도, 약물의 활성, 약물에 대한 민감도, 투여 시간, 투여 경로 및 배출 비율, 치료 기간, 동시 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 결정될 수 있다.The pharmaceutical composition may be administered to a subject in a pharmaceutically effective amount. The “pharmaceutically effective amount” means an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is dependent on the patient's type, severity, drug activity, and drug Sensitivity, time of administration, route of administration and rate of excretion, duration of treatment, factors including concomitant drugs, and other factors well known in the medical field.
상기 약학적 조성물은 개별 치료제로 투여하거나 다른 치료제와 병용하여 투여될 수 있고, 종래의 치료제와 순차적으로 또는 동시에 투여될 수 있으며, 단일 또는 다중 투여될 수 있다. 상기 요소들을 모두 고려하여 부작용 없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 바람직하며, 이는 당업자에 의해 용이하게 결정될 수 있다.The pharmaceutical composition may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered single or multiple. Taking all of the above factors into consideration, it is preferable to administer an amount capable of obtaining the maximum effect with a minimum amount without side effects, which can be easily determined by those skilled in the art.
본 발명의 다른 측면에 따르면, 로스팅(roasting) 좁은잎천선과 추출물을 유효성분으로 포함하는 면역 증진용 조성물이 제공된다.According to another aspect of the present invention, there is provided a composition for enhancing immunity comprising the extract of Roasting Narrowleaf chrysanthemum as an active ingredient.
상기 "로스팅(roasting)"은 150℃ 이상의 높은 건열을 사용해 수분을 더하지 않고 가열하는 방법이다. 상기 로스팅을 위한 가열 장치는 오븐, 가스레인지, 전자레인지 등이 될 수 있으나, 온도를 조절할 수 있는 가열 장치라면 특별히 제한되지 않는다.The "roasting" is a method of heating without adding moisture using high dry heat of 150° C. or higher. The heating device for the roasting may be an oven, a gas range, a microwave oven, etc., but is not particularly limited as long as it is a heating device capable of controlling the temperature.
상기 좁은잎천선과는 150 내지 300℃에서 로스팅할 수 있다.The narrow-leaf cheonseonwa can be roasted at 150 to 300 ℃.
구체적으로, 상기 좁은잎천선과는 150 내지 300℃에서 5 내지 50분간 로스팅될 수 있고, 바람직하게는 200 내지 250℃에서 20 내지 30분간 로스팅될 수 있다.Specifically, the narrow-leaf cheonseonwa may be roasted at 150 to 300° C. for 5 to 50 minutes, preferably at 200 to 250° C. for 20 to 30 minutes.
상기 로스팅에 의해 좁은잎천선과에 함유된 다양한 유효성분의 함량이 증감할 수 있으며, 본 발명자들은 상기 로스팅 공정에 의해 면역 증진 활성이 현저히 증대될 수 있음을 규명하였다.By the roasting, the content of various active ingredients contained in the Narrow-leaf Cheesonaceae can be increased or decreased, and the present inventors have found that the immune enhancing activity can be significantly increased by the roasting process.
이 때 상기 좁은잎천선과는 고온에서 저온으로 단계적으로 온도를 변경하면서 로스팅할 수 있고, 예컨대 상기 좁은잎천선과를 250℃에서 150℃까지 온도를 조절하면서 로스팅할 수 있다.At this time, the narrow-leaf cheonseonwa may be roasted while changing the temperature step by step from high temperature to low temperature, for example, the narrow-leaf cheonseonwa may be roasted while controlling the temperature from 250°C to 150°C.
구체적으로 상기 로스팅 과정에서 온도를 250℃에서 150℃까지 점진적으로 저하시킬 수 있고, 250℃, 200℃, 150℃에서 각각 소정의 시간 동안 로스팅할 수도 있다.Specifically, in the roasting process, the temperature may be gradually lowered from 250° C. to 150° C., and roasted at 250° C., 200° C., and 150° C. for a predetermined time, respectively.
상기 좁은잎천선과는 고온에서 저온으로 단계적으로 로스팅을 실시함으로써 면역 증진 활성이 더욱 개선될 수 있다.Immune enhancing activity can be further improved by performing the roasting in stages from high temperature to low temperature with the narrow-leaf Cheoncheongwa.
상기 로스팅(roasting) 좁은잎천선과 추출물은 열수 추출(hot water extraction)될 수 있다.The roasted Narrowleaf chrysanthemum extract may be subjected to hot water extraction.
본 발명자들은 로스팅된 좁은잎천선과를 열수 추출시킴으로써 면역 증진 활성이 현저히 증대될 수 있음을 확인하였으며, 상기 열수 추출물은 면역 증진을 위한 식품 및 의약품 제조에 유용하게 활용될 수 있다.The present inventors have confirmed that the immune enhancing activity can be remarkably increased by hot water extraction of the roasted Narrowleaf Cheonenaceae, and the hot water extract can be usefully used in the manufacture of foods and pharmaceuticals for enhancing immunity.
이하 실시예를 통해, 본 발명을 더욱 상술하나 하기 실시예에 의해 본 발명이 제한되지 아니함은 자명하다.Through the following examples, the present invention will be described in more detail, but it is obvious that the present invention is not limited by the following examples.
제조예 1 : 열수 추출물(SW1902-W) 제조Preparation Example 1: Preparation of hot water extract (SW1902-W)
파쇄한 좁은잎천선과 건조물에 100배의 정제수를 넣고 100℃에서 4시간 동안 가열한 후 여과하였다. 100-fold purified water was added to the crushed Narrow-leaf cheonseon and dried matter, heated at 100° C. for 4 hours, and then filtered.
수득한 액상 추출물을 회전감압농축기로 농축(20 brix)한 후 동결건조기를 이용하여 좁은잎천선과 추출물을 수득하였다. 분말화하여 수득한 추출물의 수율은 건조물 대비 36.5%였다.The obtained liquid extract was concentrated (20 brix) with a rotary vacuum concentrator, and then an extract of Narrowleaf chrysanthemum was obtained using a freeze dryer. The yield of the extract obtained by pulverization was 36.5% compared to the dry matter.
제조예 2 : 에탄올 추출물(SW1902-70E) 제조Preparation Example 2: Preparation of ethanol extract (SW1902-70E)
파쇄한 좁은잎천선과 건조물에 100배의 에탄올(70% v/v)를 넣고 100℃에서 4시간 동안 가열한 후 여과하였다.100 times of ethanol (70% v/v) was added to the crushed narrow-leaf chrysanthemum and dried matter, heated at 100° C. for 4 hours, and then filtered.
수득한 액상 추출물을 회전감압농축기로 농축(20 brix)한 후 동결건조기를 이용하여 좁은잎천선과 추출물을 수득하였다. 분말화하여 수득한 추출물의 수율은 건조물 대비 27.2%였다.The obtained liquid extract was concentrated (20 brix) with a rotary vacuum concentrator, and then an extract of Narrowleaf chrysanthemum was obtained using a freeze dryer. The yield of the extract obtained by pulverization was 27.2% compared to the dry matter.
제조예 3 : 로스팅 열수 추출물(SW1902-250R-W) 제조Preparation Example 3: Preparation of roasted hot water extract (SW1902-250R-W)
좁은잎천선과 건조 시료에 대한 가열처리(로스팅)를 수행하였다. 미리 충분히 예열 후 로스팅 공정을 수행하였다. Heat treatment (roasting) was performed on the narrow-leaf chrysanthemum and dried samples. After sufficiently preheating in advance, the roasting process was performed.
로스팅 온도는 250℃이었으며, 20분, 25분, 30분 시간 범위에서 바이오테크 FEC-006를 이용하여 로스팅을 수행하였다.The roasting temperature was 250° C., and roasting was performed using Biotech FEC-006 in the time ranges of 20 minutes, 25 minutes, and 30 minutes.
각 조건에서 각 로스팅된 좁은잎천선과는 동등한 수준의 활성이 나타났으나, 250℃에서 25분간 로스팅된 좁은잎천선과의 활성이 가장 우수하였으므로 해당 시료의 활성 평가를 진행하였다.In each condition, the same level of activity was shown with each roasted Narrowleaf P., but the activity was the most excellent with Narrowleaf Prunus roasted at 250°C for 25 minutes, so the activity of the sample was evaluated.
파쇄한 로스팅 좁은잎천선과 건조물에 100배의 정제수를 넣고 100℃에서 4시간 동안 가열한 후 여과하였다.100-fold purified water was added to the crushed roasted narrow-leaf cheonseon and dried, heated at 100° C. for 4 hours, and then filtered.
수득한 액상 추출물을 회전감압농축기로 농축(20 brix)한 후 동결건조기를 이용하여 좁은잎천선과 추출물을 수득하였다. 분말화하여 수득한 추출물의 수율은 건조물 대비 36.9%였다.The obtained liquid extract was concentrated (20 brix) with a rotary vacuum concentrator, and then an extract of Narrowleaf chrysanthemum was obtained using a freeze dryer. The yield of the extract obtained by pulverization was 36.9% compared to the dry matter.
제조예 4 : 로스팅 에탄올 추출물(SW1902-250R-70E) 제조Preparation Example 4: Preparation of roasted ethanol extract (SW1902-250R-70E)
제조예 3과 동일한 방법으로 로스팅을 수행하였다. 250℃에서 25분간 로스팅된 좁은잎천선과로 활성 평가를 진행하였다.Roasting was performed in the same manner as in Preparation Example 3. Activity evaluation was conducted with Narrow-leaf Cheoncheongwa roasted at 250° C. for 25 minutes.
파쇄한 로스팅 좁은잎천선과 건조물에 100배의 에탄올(70% v/v)을 넣고 100℃에서 4시간 동안 가열한 후 여과하였다.100 times of ethanol (70% v/v) was added to the crushed roasted narrow-leaf cheonseon and dried matter, heated at 100° C. for 4 hours, and then filtered.
수득한 액상 추출물을 회전감압농축기로 농축(20 brix)한 후 동결건조기를 이용하여 좁은잎천선과 추출물을 수득하였다. 분말화하여 수득한 추출물의 수율은 건조물 대비 26.2%였다.The obtained liquid extract was concentrated (20 brix) with a rotary vacuum concentrator, and then an extract of Narrowleaf chrysanthemum was obtained using a freeze dryer. The yield of the extract obtained by pulverization was 26.2% compared to the dry matter.
제조예 5 : 로스팅 열수 추출물(SW1902-200R-W) 제조Preparation Example 5: Preparation of roasted hot water extract (SW1902-200R-W)
제조예 3과 동일한 방법으로 열수 추출물을 제조하되, 로스팅 온도를 200℃로 하였다. 분말화하여 수득한 추출물의 수율은 건조물 대비 36.3%였다.A hot water extract was prepared in the same manner as in Preparation Example 3, but the roasting temperature was 200°C. The yield of the extract obtained by pulverization was 36.3% compared to the dry matter.
제조예 6 : 로스팅 에탄올 추출물(SW1902-200R-70E) 제조Preparation Example 6: Preparation of roasted ethanol extract (SW1902-200R-70E)
제조예 4과 동일한 방법으로 에탄올 추출물을 제조하되, 로스팅 온도를 200℃로 하였다. 분말화하여 수득한 추출물의 수율은 건조물 대비 26.0%였다.An ethanol extract was prepared in the same manner as in Preparation Example 4, but the roasting temperature was 200°C. The yield of the extract obtained by pulverization was 26.0% compared to the dry matter.
제조예 7 : 로스팅 열수 추출물(SW1902-R(3STEP)-W) 제조Preparation Example 7: Preparation of roasted hot water extract (SW1902-R(3STEP)-W)
제조예 3과 동일한 방법으로 열수 추출물을 제조하되, 로스팅 온도를 단계적으로 변경하였다.A hot water extract was prepared in the same manner as in Preparation Example 3, but the roasting temperature was changed in stages.
구체적으로, 로스팅 공정은 25분간 진행하되 250℃에서 8.5분, 200℃에서 8.5분, 150℃에서 8분간 단계적으로 진행하였다. Specifically, the roasting process was carried out in stages for 25 minutes, but at 250°C for 8.5 minutes, at 200°C for 8.5 minutes, and at 150°C for 8 minutes.
분말화하여 수득한 추출물의 수율은 건조물 대비 27.8%였다.The yield of the extract obtained by pulverization was 27.8% compared to the dry matter.
제조예 8 : 로스팅 에탄올 추출물(SW1902-R(3STEP)-70E) 제조Preparation Example 8: Preparation of roasted ethanol extract (SW1902-R(3STEP)-70E)
제조예 3과 동일한 방법으로 에탄올 추출물을 제조하되, 로스팅 온도를 단계적으로 변경하였다.An ethanol extract was prepared in the same manner as in Preparation Example 3, but the roasting temperature was changed in stages.
구체적으로, 로스팅 공정은 25분간 진행하되 250℃에서 8.5분, 200℃에서 8.5분, 150℃에서 8분간 단계적으로 진행하였다.Specifically, the roasting process was carried out in stages for 25 minutes, but at 250°C for 8.5 minutes, at 200°C for 8.5 minutes, and at 150°C for 8 minutes.
분말화하여 수득한 추출물의 수율은 건조물 대비 23.9%였다.The yield of the extract obtained by pulverization was 23.9% compared to the dry matter.
실험예 1 : 세포 배양Experimental Example 1: Cell Culture
세포주인 RAW264.7 macrophage cell은 American Type Culture Collection(ATCC, Manassas. USA)에서 구입하여 사용하였다.The cell line, RAW264.7 macrophage cell, was purchased from the American Type Culture Collection (ATCC, Manassas. USA).
세포의 배양을 위하여 10% Fetal Bovine Serum(FBS, Hyclone USA), 100 units/mL penicillin/Streptomycin(Hyclone, USA)이 함유된 DMEM(Dulbecco's modified of Eagle's medium, Hyclone,USA) 배지를 사용하였으며, 37℃, 5% CO2 존재하의 incubator(Thermo, Forma, Germany)에서 배양하였다.For cell culture, DMEM (Dulbecco's modified of Eagle's medium, Hyclone, USA) medium containing 10% Fetal Bovine Serum (FBS, Hyclone USA) and 100 units/mL penicillin/Streptomycin (Hyclone, USA) was used, 37 C, 5% CO 2 Incubated in an incubator (Thermo, Forma, Germany) in the presence.
실험예 2 : 독성 평가(MTT assay)Experimental Example 2: Toxicity evaluation (MTT assay)
살아있는 세포 내 미토콘드리아의 탈수소효소에 의해 Tetrazolium salt(WST-1)에서 formazan으로 변화하는 것을 이용하여, 생성된 formazan의 농도를 spectrophotometer로 측정함으로써 세포 생존률을 측정하였다.Cell viability was measured by measuring the concentration of the generated formazan with a spectrophotometer using a change from tetrazolium salt (WST-1) to formazan by mitochondrial dehydrogenase in living cells.
Ez-cytox(Dozen, Korea)을 이용하여 프로토콜에 따라 MTT assay를 수행하였다.MTT assay was performed according to the protocol using Ez-cytox (Dozen, Korea).
Raw264.7 세포를 5x103 cells/100μL로 96 well에 분주한 후 37℃, 5% CO2 배양기에서 24시간 동안 배양하였다.Raw264.7 cells were aliquoted into 96 wells at 5x10 3 cells/100 μL and cultured at 37° C., 5% CO 2 in an incubator for 24 hours.
농도별로 시료를 처리한 후 24시간 동안 배양하였다. 배양 후 10% MTT 시약이 포함된 배지로 교체하고 37℃에서 1시간 동안 반응시킨 후 450 nm 파장에서 microplate reader(Epoch, Biotek)로 측정하였다.Samples were treated for each concentration and incubated for 24 hours. After incubation, the medium was replaced with a medium containing 10% MTT reagent and reacted at 37° C. for 1 hour, followed by measurement with a microplate reader (Epoch, Biotek) at a wavelength of 450 nm.
각 시료에 대한 세포생존율에 의한 독성 평가는 3회 반복하여 측정하였으며, 그에 대한 평균값과 표준편차를 산출하였다.Toxicity evaluation by cell viability for each sample was repeated three times, and the average value and standard deviation thereof were calculated.
도 1을 참조하면, 제조예 1 내지 8의 추출물은 모든 농도(10, 100, 1000 μg/mL) 범위에서 세포 독성이 나타나지 않았다.1, the extracts of Preparation Examples 1 to 8 did not show cytotoxicity in all concentrations (10, 100, 1000 μg/mL) range.
실험예 3 : 면역 효능 평가(NO assay)Experimental Example 3: Immune efficacy evaluation (NO assay)
Nitric Oxide(NO)의 농도 측정을 위해 그리스 반응(griess reaction)을 이용하여 540nm에서 흡광도를 측정하였다.Absorbance was measured at 540 nm using a grease reaction to measure the concentration of nitric oxide (NO).
N-(1-naphthyl)-ethylenediamine, sulfanilamide, 및 NO2 -가 반응하여 azo coupling 을 이루는데, 두 개의 링 형태가 540nm의 파장에서 최대 흡광도 값을 가진다.N-(1-naphthyl)-ethylenediamine, sulfanilamide, and NO 2 - react to form azo coupling, and the two rings have the maximum absorbance value at a wavelength of 540 nm.
Raw264.7 세포를 5x104 cells/250μL로 24 well plate 에 분주하고 37℃, 5% CO2 배양기에서 24시간 동안 배양하였다.Raw264.7 cells were aliquoted in a 24 well plate at 5x10 4 cells/250 μL and cultured at 37° C., 5% CO 2 in an incubator for 24 hours.
시료를 농도별로 처리하고 48시간 동안 CO2 배양기에 배양하였다. 세포배양 상등액 50μL를 취해 N1 buffer 50μL를 혼합하고 5 내지 10분 상온에 반응시켰다. N2 buffer 50μL를 혼합하여 10분간 반응시킨 후 microplate reader(Epoch, Biotek)를 이용하여 560nm 흡광도를 측정하였다.Samples were treated by concentration and incubated in a CO 2 incubator for 48 hours. 50 μL of the cell culture supernatant was taken, mixed with 50 μL of N1 buffer, and reacted at room temperature for 5 to 10 minutes. After mixing 50 μL of N2 buffer and reacting for 10 minutes, absorbance at 560 nm was measured using a microplate reader (Epoch, Biotek).
동일한 방법으로 3회 측정하였으며, 그에 대한 평균값과 표준편차를 구하였다.It was measured three times in the same way, and the average value and standard deviation thereof were obtained.
도 2를 참조하면, 좁은잎천선과 열수 추출물(제조예 1)의 경우 1000 μg/mL에서 NO 농도가 대조군 대비 약 49% 증가하였으며, 좁은잎천선과 에탄올 추출물(제조예 2)의 경우 대조군 대비 유의미한 증가는 나타나지 않았다.Referring to FIG. 2 , in the case of the Narrow-leaf chrysanthemum and hot water extract (Preparation Example 1), the NO concentration was increased by about 49% compared to the control at 1000 μg/mL, and in the case of the Narrow-leaved chrysanthemum and the ethanol extract (Preparation Example 2) compared to the control group No significant increase was observed.
250℃에서 로스팅된 좁은잎천선과 열수 추출물(제조예 3)의 경우 100 μg/mL에서 NO 농도가 대조군 대비 약 44%, 1000 μg/mL에서 약 115% 증가하였으며, 에탄올 추출물(제조예 4)의 경우 1000 μg/mL에서 약 21% 증가하였다.In the case of the Narrowleaf chrysanthemum and hot water extract (Preparation Example 3) roasted at 250 ° C, the NO concentration at 100 μg/mL increased by about 44% compared to the control and about 115% at 1000 μg/mL, and the ethanol extract (Preparation Example 4) In the case of , it increased by about 21% at 1000 μg/mL.
200℃에서 로스팅된 좁은잎천선과 열수 추출물(제조예 5)의 경우 1000 μg/mL에서 NO 농도가 대조군 대비 약 45% 증가하였으며, 에탄올 추출물(제조예 6)의 경우 대조군 대비 유의미한 증가는 나타나지 않았다.In the case of the Narrowleaf chrysanthemum roasted at 200 ° C. and the hot water extract (Preparation Example 5), the NO concentration increased by about 45% compared to the control at 1000 μg/mL, and in the case of the ethanol extract (Preparation Example 6), there was no significant increase compared to the control. .
단계적으로 온도를 달리하여 로스팅한 좁은잎천선과 열수 추출물(제조예 7)의 경우 100 μg/mL에서 NO 농도가 대조군 대비 약 51%, 1000 μg/mL에서 약 96% 증가하였으며, 에탄올 추출물(제조예 8)의 경우 대조군 대비 유의미한 증가는 나타나지 않았다.In the case of the Narrowleaf chrysanthemum and hot water extract (Preparation Example 7) roasted at different temperatures in stages, the NO concentration at 100 µg/mL increased by about 51% compared to the control group and by about 96% at 1000 µg/mL, and the ethanol extract (Preparation Example 7) In the case of Example 8), there was no significant increase compared to the control group.
상기 결과는 좁은잎천선과가 열수 추출시 면역 증진 효과가 우수할 뿐만 아니라, 로스팅에 의해서 면역 증진 활성이 현저하게 증대될 수 있음을 시사한다.The above results suggest that Narrowleaf Cheonenaceae not only has an excellent immune enhancing effect during hot water extraction, but also that the immune enhancing activity can be remarkably increased by roasting.
실험예 4 : 면역 효능 평가(TNF-a 생성량 평가)Experimental Example 4: Immune efficacy evaluation (TNF-a production amount evaluation)
면역 증가 효능을 알아보기 위해 Raw264.7 세포를 5x104 cells/250μL로 24 well plate 에 분주하고 37℃, 5% CO2 배양기에서 24시간 동안 배양하였다.In order to examine the efficacy of increasing immunity, Raw264.7 cells were aliquoted at 5x10 4 cells/250 μL in a 24 well plate and cultured at 37° C., 5% CO 2 in an incubator for 24 hours.
시료를 농도별로 처리하고 4시간동안 CO2 배양기에 배양한 후 세포 부유액을 원심 분리하여 세포들은 침전시켜 상층액을 수집하였다.The samples were treated by concentration and incubated in a CO 2 incubator for 4 hours, then the cell suspension was centrifuged to precipitate the cells, and the supernatant was collected.
상층액 내 TNF-a 생성량을 ELISA kit(R&D systems Inc., Minneapolis, MN, USA를 이용하여 사용자 매뉴얼에 기재된 방법대로 정량하였다.The amount of TNF-a production in the supernatant was quantified according to the method described in the user's manual using an ELISA kit (R&D systems Inc., Minneapolis, MN, USA).
도 3을 참조하면, 좁은잎천선과 열수 추출물(제조예 1)의 경우 100 μg/mL에서 대조군 대비 TNF-a 양이 약 101 pg/mL 증가하였으며, 1000 μg/mL에서 대조군 대비 약 296 pg/mL 증가하였다.Referring to FIG. 3, in the case of the Narrowleaf chrysanthemum extract (Preparation Example 1), the amount of TNF-a increased by about 101 pg/mL compared to the control at 100 μg/mL, and at 1000 μg/mL, about 296 pg/mL compared to the control. mL increased.
반면, 좁은잎천선과 에탄올 추출물(제조예 2)의 경우 대조군 대비 TNF-a 생성량의 유의미한 증가는 나타나지 않았다.On the other hand, in the case of Narrow-leaf Cheoncheon and ethanol extract (Preparation Example 2), there was no significant increase in the amount of TNF-a production compared to the control group.
250℃에서 로스팅된 좁은잎천선과 열수 추출물(제조예 3)의 경우 100 μg/mL에서 대조군 대비 TNF-a 양이 약 306 pg/mL, 1000 μg/mL에서 대조군 대비 약 468 pg/mL 증가하였으며, 에탄올 추출물(제조예 4)의 경우 100 μg/mL에서 대조군 대비 TNF-a 양이 약 68 pg/mL, 1000 μg/mL에서 대조군 대비 약 246 pg/mL 증가하였다.The amount of TNF-a was increased by about 306 pg/mL compared to the control at 100 μg/mL, and by about 468 pg/mL compared to the control at 1000 μg/mL in the case of the Narrowleaf chrysanthemum and hot water extract (Preparation Example 3) roasted at 250 ° C. , In the case of the ethanol extract (Preparation Example 4), the amount of TNF-a was increased by about 68 pg/mL compared to the control at 100 μg/mL, and by about 246 pg/mL compared to the control at 1000 μg/mL.
200℃에서 로스팅된 좁은잎천선과 열수 추출물(제조예 5)의 경우 100 μg/mL에서 대조군 대비 TNF-a 양이 약 202 pg/mL, 1000 μg/mL에서 대조군 대비 약 371 pg/mL 증가하였으며, 에탄올 추출물(제조예 6)의 경우 대조군 대비 TNF-a 생성량의 유의미한 증가는 나타나지 않았다.In the case of the Narrowleaf chrysanthemum and hot water extract (Preparation Example 5) roasted at 200 °C, the amount of TNF-a was increased by about 202 pg/mL compared to the control at 100 μg/mL, and by about 371 pg/mL compared to the control at 1000 μg/mL. , In the case of the ethanol extract (Preparation Example 6), there was no significant increase in the amount of TNF-a production compared to the control group.
단계적으로 온도를 달리하여 로스팅한 좁은잎천선과 열수 추출물(제조예 7)의 경우 100 μg/mL에서 대조군 대비 TNF-a 양이 약 307 pg/mL, 1000 μg/mL에서 대조군 대비 약 476 pg/mL 증가하였으며, 에탄올 추출물(제조예 4)의 경우 1000 μg/mL에서 대조군 대비 TNF-a 양이 약 180 pg/mL 증가하였다.In the case of the Narrowleaf chrysanthemum and hot water extract (Preparation Example 7) roasted at different temperatures in stages, the amount of TNF-a was about 307 pg/mL compared to the control at 100 μg/mL, and about 476 pg/mL compared to the control at 1000 μg/mL. mL was increased, and in the case of the ethanol extract (Preparation Example 4), the amount of TNF-a was increased by about 180 pg/mL compared to the control at 1000 μg/mL.
상기 좁은잎천선과는 로스팅 처리하였을 때 특히 낮은 농도에서도 높은 수준의 면역 증진 활성이 확인되었다.A high level of immune-enhancing activity was confirmed even at a particularly low concentration when the Narrowleaf Chrysanthemum was roasted.
상기 결과는 좁은잎천선과가 열수 추출시 면역 증진 효과가 우수할 뿐만 아니라, 로스팅에 의해서 면역 증진 활성이 현저하게 증대될 수 있음을 시사한다.The above results suggest that Narrowleaf Cheonenaceae not only has an excellent immune enhancing effect during hot water extraction, but also that the immune enhancing activity can be remarkably increased by roasting.
전술한 본 발명의 설명은 예시를 위한 것이며, 본 발명이 속하는 기술분야의 통상의 지식을 가진 자는 본 발명의 기술적 사상이나 필수적인 특징을 변경하지 않고서 다른 구체적인 형태로 쉽게 변형이 가능하다는 것을 이해할 수 있을 것이다. 그러므로 이상에서 기술한 실시예들은 모든 면에서 예시적인 것이며 한정적이 아닌 것으로 이해해야만 한다. 예를 들어, 단일형으로 설명되어 있는 각 구성 요소는 분산되어 실시될 수도 있으며, 마찬가지로 분산된 것으로 설명되어 있는 구성 요소들도 결합된 형태로 실시될 수 있다.The above description of the present invention is for illustration, and those of ordinary skill in the art to which the present invention pertains can understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. For example, each component described as a single type may be implemented in a dispersed form, and likewise components described as distributed may be implemented in a combined form.
본 발명의 범위는 후술하는 청구범위에 의하여 나타내어지며, 청구범위의 의미 및 범위 그리고 그 균등 개념으로부터 도출되는 모든 변경 또는 변형된 형태가 본 발명의 범위에 포함되는 것으로 해석되어야 한다.The scope of the present invention is indicated by the following claims, and all changes or modifications derived from the meaning and scope of the claims and their equivalents should be construed as being included in the scope of the present invention.
Claims (7)
상기 좁은잎천선과는 150 내지 300℃에서 로스팅한 조성물.4. The method of claim 3,
The composition roasted at 150 to 300 ℃ with the narrow leaf cheonseonwa.
상기 좁은잎천선과는 250℃에서 150℃까지 온도를 조절하면서 로스팅한 조성물.4. The method of claim 3,
The composition roasted while controlling the temperature from 250 ℃ to 150 ℃ with the narrow leaf cheonseonwa.
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