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KR102224051B1 - Composition for enhancing neutralizing antibody generation rate comprising Zingiber officinale extract as effective component - Google Patents

Composition for enhancing neutralizing antibody generation rate comprising Zingiber officinale extract as effective component Download PDF

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KR102224051B1
KR102224051B1 KR1020190039630A KR20190039630A KR102224051B1 KR 102224051 B1 KR102224051 B1 KR 102224051B1 KR 1020190039630 A KR1020190039630 A KR 1020190039630A KR 20190039630 A KR20190039630 A KR 20190039630A KR 102224051 B1 KR102224051 B1 KR 102224051B1
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유수성
이호영
김지현
심은형
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한국 한의학 연구원
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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00Medicinal preparations containing antigens or antibodies
    • A61K2039/555Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55588Adjuvants of undefined constitution

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Abstract

본 발명은 건강 추출물을 유효성분으로 포함하는 중화항체 생성율 증진용 조성물에 관한 것으로, 본 발명의 건강 추출물을 투여한 후, 백신을 접종한 동물모델은 항원 특이 항체 형성이 증진되었고, 항암제에 의해 저하되었던 항원 자극에 의해 생성되는 사이토카인의 생성 비율이 회복되며, 백신 접종 후, H1N1 및 H3N2 항원에 대한 항체 생성율이 증진되는 효과가 있으므로, 본 발명의 조성물을 독감백신 접종 전에 면역력인 낮아진 환자 또는 노인들에게 적용하면 항체 생성율을 증진시킬 수 있을 것으로 기대된다.The present invention relates to a composition for enhancing the production rate of neutralizing antibodies comprising a healthy extract as an active ingredient, and after administering the healthy extract of the present invention, in the animal model inoculated with the vaccine, the formation of antigen-specific antibodies was enhanced, and decreased by anticancer agents. Since the rate of cytokine production generated by antigen stimulation is restored, and the rate of antibody production against H1N1 and H3N2 antigens is increased after vaccination, the composition of the present invention is used in patients with lowered immunity before inoculation with flu vaccine or elderly people. It is expected to be able to improve the antibody production rate when applied to them.

Description

건강 추출물을 유효성분으로 포함하는 중화항체 생성율 증진용 조성물{Composition for enhancing neutralizing antibody generation rate comprising Zingiber officinale extract as effective component} Composition for enhancing neutralizing antibody generation rate comprising Zingiber officinale extract as effective component

본 발명은 건강 추출물을 유효성분으로 포함하는 중화항체 생성율 증진용 조성물에 관한 것이다.The present invention relates to a composition for enhancing the production rate of neutralizing antibodies comprising a health extract as an active ingredient.

독감 바이러스는 매년 전세계적으로 발생하여 호흡기 질환을 일으키는데, 이 질환은 면역 능력이 약한 어린이와 노인들에 대해 치사율이 높고, 합병증으로 폐렴(pneumonia), 심폐질환(cardiopulmonary disease) 등의 기관지 질환들을 수반하는 경우 치사율이 더욱 높아져 국민 보건에 미치는 영향이 크다고 할 수 있다. The flu virus occurs worldwide every year and causes respiratory disease, which has a high mortality rate in children and elderly people with weak immune systems, and complications include bronchial diseases such as pneumonia and cardiopulmonary disease. If it does, it can be said that the mortality rate is even higher, which has a great impact on public health.

특히 1918년에 약 2,000 만명의 사망자를 낸 스페인 독감 바이러스와 같이, 10 내지 20년 주기로 전세계으로 출현하는(pandemic) 독감 바이러스는 매우 치명적인데, 60년대의 홍콩 독감 바이러스 이후 이러한 독감 바이러스가 발생하지 않았다는 점을 고려하면, 가까운 미래에 치명적인 독감 바이러스의 출현이 예상되고 있다. 이러한 독감 바이러스의 예방을 위해, 세계 보건 기구(WHO)는 통계를 토대로 1 년후에 발생가능한 독감 바이러스를 세 유형별로 미리 선정하여 고시함으로써 백신이 보급되도록 하고 있다. 하지만 아직까지 백신을 접종한 후, 항체 형성 여부에 대한 검증은 이루어지지 않고 있다. 대부분의 건강한 사람은 항체 형성이 잘 이루졌을 것으로 예상되지만 면역력이 낮아진 환자군 또는 노인들의 경우는 항체 형성이 매우 낮은 것으로 알려져 있다. In particular, the pandemic flu virus, which occurs every 10 to 20 years, is very deadly, such as the Spanish flu virus, which killed about 20 million people in 1918. It is said that this flu virus has not occurred since the Hong Kong flu virus in the 60s. Considering that, the emergence of a deadly flu virus is expected in the near future. To prevent this flu virus, the World Health Organization (WHO) is pre-selecting and notifying the three types of flu viruses that can occur after one year based on statistics so that the vaccine is distributed. However, after vaccination, there is still no verification of the formation of antibodies. Antibody formation is expected to be well-formed in most healthy people, but it is known that antibody formation is very low in patients with low immunity or in the elderly.

이와 같이 바이러스 감염에 취약한 환자 또는 노인이 예방접종을 통해 항체 생성률이 낮다는 문제점이 제기되고 있으나, 아직까지는 이를 극복하기 위한 방안 마련은 거의 없는 수준이다. As such, a problem has been raised that patients or the elderly who are susceptible to viral infection have a low antibody production rate through vaccination, but there is little to no plan to overcome this problem.

한편, 건강(Zingiber officinale)은 아열대 및 열대성 다년생 식물로서 근경을 주로 식용하며, 그 특유한 향기와 매운 맛으로 인하여 오랫동안 향신료로서 사용되고 있다. 생강의 생리 활성 성분이 항균작용, 항염작용, 혈청 콜레스테롤 저하 효과, 항산화 작용을 나타내는 것으로 알려져 있다. 건강으로부터 분리한 올레오레진(oleoresin), 진저롤(gingerol) 및 쇼가올(shogaol)이 자연살해세포 기능을 활성화시켜 면역능 증진에 효과가 있다는 것으로 알려져 있고, [한국영양과학회지 37(1); 23~30, 2004]에 생강추출물 투여가 마우스 면역세포 활성에 미치는 영향이 개시되어 있으며, 한국등록특허 제1324926호에 생강 추출물 제조방법 및 이에 따른 생강 추출물이 개시되어 있으나, 본 발명의 건강 추출물을 유효성분으로 포함하는 중화항체 생성율 증진용 조성물에 대해 개시된 바는 없다. Meanwhile, health ( Zingiber officinale ) is a subtropical and tropical perennial plant, mainly edible with rhizomes, and has been used as a spice for a long time due to its peculiar aroma and spicy taste. It is known that the physiologically active ingredients of ginger have antibacterial, anti-inflammatory, serum cholesterol lowering, and antioxidant effects. It is known that oleoresin, gingerol, and shogaol isolated from health are effective in enhancing immunity by activating natural killer cell functions, [Journal of the Korean Society of Nutrition 37(1); 23-30, 2004] discloses the effect of administration of ginger extract on mouse immune cell activity, and Korean Patent No. 1324926 discloses a method for preparing a ginger extract and a ginger extract according thereto, but the health extract of the present invention There is no disclosure about the composition for enhancing the production rate of neutralizing antibodies containing as an active ingredient.

본 발명은 상기와 같은 요구에 의해 도출된 것으로서, 본 발명은 건강 추출물을 유효성분으로 포함하는 중화항체 생성율 증진용 조성물을 제공하고, 본 발명의 유효성분인 건강 추출물을 투여한 후, 백신을 접종한 동물모델은 항원 특이 항체 형성이 증진되었고, 항암제에 의해 저하되었던 항원 자극에 의해 생성되는 사이토카인의 생성 비율이 회복되며, 백신 접종 후, H1N1 및 H3N2 항원에 대한 항체 생성율이 증진되는 효과가 있다는 것을 확인함으로써, 본 발명을 완성하였다.The present invention is derived from the above requirements, the present invention provides a composition for enhancing the production rate of neutralizing antibodies comprising a health extract as an active ingredient, and after administering the health extract, the active ingredient of the present invention, vaccination One animal model showed that the formation of antigen-specific antibodies was enhanced, the rate of cytokine production, which was lowered by the anticancer drug, was restored, and the rate of antibody production against H1N1 and H3N2 antigens after vaccination was enhanced. By confirming this, the present invention was completed.

상기 목적을 달성하기 위하여, 본 발명은 건강 추출물을 유효성분으로 포함하는 독감 백신 접종에 의해 형성되는 중화항체 생성율 증진용 건강기능식품 조성물을 제공한다.In order to achieve the above object, the present invention provides a health functional food composition for enhancing the production rate of neutralizing antibodies formed by flu vaccination comprising a health extract as an active ingredient.

또한, 본 발명은 건강 추출물을 유효성분으로 포함하는 독감 백신 접종에 의해 형성되는 중화항체 생성율 증진용 약학 조성물을 제공한다.In addition, the present invention provides a pharmaceutical composition for enhancing the production rate of neutralizing antibodies formed by flu vaccination comprising a health extract as an active ingredient.

또한, 본 발명은 건강 추출물을 유효성분으로 포함하는 중화항체 생성율 증진용 독감 백신 보조제를 제공한다.In addition, the present invention provides a flu vaccine adjuvant for enhancing the production rate of neutralizing antibodies comprising a health extract as an active ingredient.

본 발명은 건강 추출물을 유효성분으로 포함하는 중화항체 생성율 증진용 조성물에 관한 것으로, 본 발명의 건강 추출물을 투여한 후, 백신을 접종한 동물모델은 항원 특이 항체 형성이 증진되었고, 항암제에 의해 저하되었던 항원 자극에 의해 생성되는 사이토카인의 생성 비율이 회복되며, 백신 접종 후, H1N1 및 H3N2 항원에 대한 항체 생성율이 증진되는 효과가 있다.The present invention relates to a composition for enhancing the production rate of neutralizing antibodies comprising a healthy extract as an active ingredient, and after administering the healthy extract of the present invention, in the animal model inoculated with the vaccine, the formation of antigen-specific antibodies was enhanced, and decreased by anticancer agents. The rate of production of cytokines generated by antigen stimulation is restored and, after vaccination, the rate of antibody production against H1N1 and H3N2 antigens is enhanced.

도 1은 본 발명의 건강 추출물의 투여에 의해 IgG 및 IgG1의 생성이 증진된 것을 확인한 결과이다. *은 항암제를 투여한 군에 비해 본 발명의 항암제 및 건강 추출물을 투여한 군에서의 IgG 및 IgG1의 생성이 통계적으로 유의미하게 증가하였다는 것으로 p<0.05이다.
도 2는 in vitro 항원 자극에 대한 IFN-γ 및 IL-4의 함량 변화를 확인한 결과로, *은 아무것도 투여하지 않은 대조군에 비해 항암제 또는 항암제와 건강추출물을 투여한 군에서의 IFN-γ 및 IL-4의 함량이 통계적으로 유의미하게 감소하였다는 것으로, p<0.05이다.
도 3은 in vitro 항원 자극에 대한 IFN-γ 및 IL-4의 비율 변화를 확인한 결과로, *은 항암제 투여군에 비해 항암제와 건강추출물을 투여한 군에서의 IFN-γ/IL-4의 비율이 통계적으로 유의미하게 증가하였다는 것으로, p<0.05이다.
도 4는 IFN-γ/IL-4의 비율과 IgG 생성량과의 관계를 확인한 결과이다.
1 is a result of confirming that the production of IgG and IgG1 was enhanced by administration of the health extract of the present invention. * Indicates that the production of IgG and IgG1 in the group administered with the anticancer agent and healthy extract of the present invention was statistically significantly increased compared to the group administered with the anticancer agent, p<0.05.
2 is a result of confirming the change in the content of IFN-γ and IL-4 upon in vitro antigen stimulation, * indicates IFN-γ and IL in the group administered with an anticancer agent or an anticancer agent and a health extract compared to the control group to which nothing was administered. The content of -4 was statistically significantly decreased, and p<0.05.
3 is a result of confirming the change in the ratio of IFN-γ and IL-4 to in vitro antigen stimulation, * indicates the ratio of IFN-γ/IL-4 in the group administered with the anticancer agent and the health extract compared to the group administered with the anticancer agent. It was a statistically significant increase, and p<0.05.
4 is a result of confirming the relationship between the ratio of IFN-γ/IL-4 and the amount of IgG produced.

본 발명은 건강 추출물을 유효성분으로 포함하는 독감 백신 접종에 의해 형성되는 중화항체 생성율 증진용 건강기능식품 조성물에 관한 것이다.The present invention relates to a health functional food composition for enhancing the production rate of neutralizing antibodies formed by flu vaccination comprising a health extract as an active ingredient.

상기 건강 추출물은 하기의 단계를 포함하는 방법에 의해 제조할 수 있으나, 이에 한정하지 않는다:The health extract may be prepared by a method comprising the following steps, but is not limited thereto:

(1) 건강에 추출용매를 가하여 추출하는 단계;(1) extracting by adding an extraction solvent to health;

(2) 단계 (1)의 추출물을 여과하는 단계; 및 (2) filtering the extract of step (1); And

(3) 단계 (2)의 여과한 추출물을 감압 농축하고 건조하여 추출물을 제조하는 단계. (3) preparing an extract by concentrating the filtered extract of step (2) under reduced pressure and drying it.

상기 단계 (1)에서 추출용매는 물, C1~C4의 저급 알코올 또는 이들의 혼합물 중에서 선택하는 것이 바람직하며, 더 바람직하게는 에탄올이고 더욱더 바람직하게는 70%(v/v) 에탄올지만 이에 한정하지 않는다.In the step (1), the extraction solvent is preferably selected from water, a lower alcohol of C 1 to C 4 or a mixture thereof, more preferably ethanol, and even more preferably 70% (v/v) ethanol. Not limited.

상기 제조방법에 있어서, 추출방법은 여과법, 열수 추출, 침지 추출, 환류 냉각 추출 및 초음파 추출 등의 당 업계에 공지된 모든 통상적인 방법을 이용할 수 있다. 상기 추출용매는 건조된 건강 중량의 1~20배 첨가하여 추출하는 것이 바람직하며, 더 바람직하게는 5~15배 첨가하는 것이다. 추출온도는 4 내지 110℃인 것이 바람직하며, 더 바람직하게는 82±2℃이지만, 이에 한정하지 않는다. 또한, 추출시간은 0.5~10시간인 것이 바람직하며, 0.5~5시간이 더욱 바람직하나 이에 한정하지 않는다. 상기 방법에 있어서, 단계 (3)의 감압농축은 진공 감압 농축기 또는 진공회전증발기를 이용하는 것이 바람직하나 이에 한정하지 않는다. 또한, 건조는 감압건조, 진공건조, 비등건조, 분무 건조 또는 동결 건조하는 것이 바람직하나 이에 한정하지 않는다.In the above manufacturing method, the extraction method may use all conventional methods known in the art such as filtration, hot water extraction, immersion extraction, reflux cooling extraction, and ultrasonic extraction. The extraction solvent is preferably extracted by adding 1 to 20 times the weight of the dried healthy, more preferably 5 to 15 times. The extraction temperature is preferably 4 to 110°C, more preferably 82±2°C, but is not limited thereto. In addition, the extraction time is preferably 0.5 to 10 hours, more preferably 0.5 to 5 hours, but is not limited thereto. In the above method, the vacuum concentration in step (3) is preferably a vacuum vacuum concentrator or a vacuum rotary evaporator, but is not limited thereto. In addition, the drying is preferably vacuum drying, vacuum drying, boiling drying, spray drying, or freeze drying, but is not limited thereto.

상기 중화항체는 H1N1 또는 H3N2의 A형 독감바이러스 감염에 대한 항체 또는 B/Phuket 또는 B/Bristone의 B형 독감바이러스 감염에 대한 항체인 것이 바람직하지만 이에 한정하지 않는다. The neutralizing antibody is preferably an antibody against H1N1 or H3N2 type A flu virus infection or B/Phuket or B/Bristone antibody against type B flu virus infection, but is not limited thereto.

상기 조성물은 면역력이 저하된 암환자 또는 60세 이상의 노인에게 독감 백신 접종 전에 투여하기 위한 것임이 바람직하지만 이에 한정하는 것은 아니다. 상기 투여는 백신 접종 1개월 전 내지 10일 전에 투여하는 것이 바람직하지만 이에 한정하지 않는다. 투여는 경구투여가 바람직하지만 이에 제한하는 것은 아니다.The composition is preferably for administration to a cancer patient with reduced immunity or an elderly person aged 60 years or older prior to inoculation of the flu vaccine, but is not limited thereto. The administration is preferably administered 1 month to 10 days before vaccination, but is not limited thereto. Administration is preferably oral administration, but is not limited thereto.

상기 건강기능식품 조성물은 분말, 과립, 환, 정제, 캡슐, 캔디, 시럽 및 음료 중에서 선택된 어느 하나의 제형으로 제조되는 것이 바람직하지만 이에 한정하지 않고, 식품의 성분으로 첨가하여 제조될 수 있으며, 통상적인 방법에 따라 적절하게 제조될 수 있다. 본 발명의 건강 추출물을 첨가할 수 있는 식품의 일례로는 육류, 소시지, 빵, 초콜릿, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알코올 음료 및 비타민 복합제 중에서 선택된 어느 하나의 형태일 수 있으며, 통상적인 의미에서의 건강기능식품을 모두 포함한다.The health functional food composition is preferably prepared in any one formulation selected from powders, granules, pills, tablets, capsules, candy, syrup, and beverages, but is not limited thereto, and may be prepared by adding it as an ingredient of food, and is usually It can be suitably manufactured according to the phosphorus method. Examples of foods to which the health extract of the present invention can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, It may be in any one form selected from tea, drink, alcoholic beverage, and vitamin complex, and includes all health functional foods in the usual sense.

본 발명의 건강기능식품 조성물은 여러 가지 영양제, 비타민, 광물(전해질), 합성 및 천연 풍미제, 착색제 및 증진제(치즈, 초콜릿 등), 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. 그 밖에 천연 과일 주스 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 본 발명의 건강기능식품 조성물은 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상기 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드, 말토스, 슈크로스와 같은 디사카라이드, 및 덱스트린, 사이클로덱스트린과 같은 폴리사카라이드, 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 감미제로서는 타우마틴, 스테비아 추출물과 같은 천연 감미제나, 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다.The health functional food composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), synthetic and natural flavors, colorants and enhancers (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective properties. It may contain colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonates used in carbonated beverages, and the like. In addition, it may contain pulp for the manufacture of natural fruit juices and vegetable beverages. These components may be used independently or in combination. The health functional food composition of the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients. The natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as taumatin and stevia extract, and synthetic sweeteners such as saccharin and aspartame can be used.

또한, 본 발명은 건강 추출물을 유효성분으로 포함하는 독감 백신 접종에 의해 형성되는 중화항체 생성율 증진용 약학 조성물에 관한 것이다.In addition, the present invention relates to a pharmaceutical composition for enhancing the production rate of neutralizing antibodies formed by flu vaccination comprising a health extract as an active ingredient.

본 발명의 약학 조성물은 상기 건강 추출물 이외에 추가로 담체, 부형제 또는 희석제를 더 포함할 수 있고, 본 발명의 약학 조성물은 경구 또는 비경구로 투여될 수 있으며, 비 경구 투여 시 피부 외용 또는 복강 내, 직장, 정맥, 근육, 피하, 자궁 내 경막 또는 뇌혈관 내 주사 방식을 선택하는 것이 바람직하다.The pharmaceutical composition of the present invention may further include a carrier, excipient, or diluent in addition to the health extract, and the pharmaceutical composition of the present invention may be administered orally or parenterally. , Intravenous, intramuscular, subcutaneous, intrauterine dura mater or cerebrovascular injection method is preferred.

본 발명의 약학 조성물은 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제될 수 있다. 경구투여를 위한 고형 제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형 제제는 하나 이상의 화합물에 적어도 하나 이상의 부형제 예를 들면, 전분, 탄산칼슘, 수크로오스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제된다. 또한, 단순한 부형제 이외에 스테아린산 마그네슘, 탈크 등과 같은 윤활제들도 사용된다. 경구 투여를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순 희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성 용제, 현탁제, 유제, 동결건조제제, 좌제가 포함된다. 비수성 용제 및 현탁 용제로는 프로필렌글리콜(propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈tween) 61, 카카오지, 라우린지, 글리세로 젤라틴 등이 사용될 수 있다.The pharmaceutical composition of the present invention may be prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient in one or more compounds, such as starch, calcium carbonate, sucrose, or lactose ( lactose), gelatin, etc. In addition, in addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, liquid solutions, emulsions, syrups, etc.In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweetening agents, fragrances, and preservatives may be included. have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, and suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. As a base for suppositories, witepsol, macrogol, twin tween) 61, cacao butter, laurin paper, glycero gelatin, and the like may be used.

본 발명에 따른 조성물은 약제학적으로 유효한 양으로 투여한다. 본 발명에 있어서, '약제학적으로 유효한 양'은 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분한 양을 의미하며, 유효용량 수준은 환자의 질환의 종류, 중증도, 약물의 활성, 약물에 대한 민감도, 투여 시간, 투여 경로 및 배출 비율, 치료기간, 동시 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 결정될 수 있다. 본 발명의 조성물은 개별 치료제로 투여하거나 다른 치료제와 병용하여 투여될 수 있고 종래의 치료제와는 순차적 또는 동시에 투여될 수 있으며, 단일 또는 다중 투여될 수 있다. 상기한 요소들을 모두 고려하여 부작용없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하며, 이는 당업자에 의해 용이하게 결정될 수 있다.The composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention,'pharmaceutically effective amount' means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is the type of disease, severity, and drug activity of the patient. , Sensitivity to drugs, time of administration, route of administration and rate of excretion, duration of treatment, factors including drugs used concurrently, and other factors well known in the medical field. The composition of the present invention may be administered as an individual therapeutic agent or administered in combination with other therapeutic agents, may be administered sequentially or simultaneously with a conventional therapeutic agent, and may be administered single or multiple. It is important to administer an amount capable of obtaining the maximum effect in a minimum amount without side effects in consideration of all the above factors, and this can be easily determined by a person skilled in the art.

본 발명의 조성물의 투여량은 환자의 체중, 연령, 성별, 건강상태, 식이, 투여시간, 투여방법, 배설률 및 질환의 중증도에 따라 그 범위가 다양하게 사용할 수 있다.The dosage of the composition of the present invention can be used in various ranges according to the patient's weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of the disease.

또한, 본 발명은 건강 추출물을 유효성분으로 포함하는 중화항체 생성율 증진용 독감 백신 보조제에 관한 것이다.In addition, the present invention relates to a flu vaccine adjuvant for enhancing the production rate of neutralizing antibodies comprising a health extract as an active ingredient.

상기 독감 백신 보조제는 건강 추출물에 추가로 동일 또는 유사한 기능을 나타내는 유효성분을 1종 이상을 함유할 수 있다. 상기 독감 백신 보조제는 임상 투여 시에 경구 또는 비경구로 투여가 가능하며, 비경구 투여 시 복강 내 주사, 직장 내 주사, 피하주사, 정맥주사, 근육 내 주사, 자궁 내 경막주사, 뇌혈관 내 주사 또는 흉부 내 주사에 의해 투여될 수 있고, 일반적인 의약품 제제의 형태로 사용될 수 있다. The flu vaccine adjuvant may contain one or more active ingredients exhibiting the same or similar function in addition to the health extract. The flu vaccine adjuvant can be administered orally or parenterally during clinical administration, and when administered parenterally, intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection, intrauterine dural injection, cerebrovascular injection, or It can be administered by intrathoracic injection, and can be used in the form of a general pharmaceutical formulation.

본 발명의 독감 백신 보조제는 실제 임상 투여 시에 비경구의 여러 가지 제형으로 투여될 수 있는데, 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조제제, 좌제가 포함된다. 비수성용제, 현탁용제로는 프로필렌글리콜(Propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로제라틴 등이 사용될 수 있다.The flu vaccine adjuvant of the present invention can be administered in various parenteral formulations at the time of actual clinical administration.In the case of formulation, diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants are used. It is prepared by doing. Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerogelatin, and the like may be used.

이하, 실시예를 이용하여 본 발명을 더욱 상세하게 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로 본 발명의 범위가 이들에 의해 제한되지 않는다는 것은 당해 기술분야에서 통상의 지식을 가진 자에게 있어 자명한 것이다. Hereinafter, the present invention will be described in more detail using examples. These examples are for illustrative purposes only, and it is obvious to those of ordinary skill in the art that the scope of the present invention is not limited thereto.

실시예Example 1. 건강 추출물의 제조 1. Preparation of healthy extract

건강 무게 대비 10배의 70%(v/v) 에탄올을 첨가하여 82±2℃에서 3시간 동안 환류 추출을 하였다. 그 후 거름망 필터 후, 솜 필터를 진행하였다. 필터된 용액은 동결건조하여 분말화하였다.Reflux extraction was performed at 82±2° C. for 3 hours by adding 70% (v/v) ethanol 10 times the weight of healthy weight. Thereafter, after the sieve filter, a cotton filter was performed. The filtered solution was lyophilized to powder.

[동물모델 및 재료][Animal model and materials]

본 발명의 실험동물은 Balb/c 8주령 암컷 마우스를 오리엔트 바이오로부터 구입하였으며, 상기 시클로포스파미드(cyclophosphamide, Sigma) 150mg/kg을 2일 간격으로 총 3회 복강 내 주사하였다. 실험군은 마지막 항암제 투여 후, 3일째부터 10일 동안 500mg/kg의 건강추출물을 경구투여하였고, 대조군은 증류수를 경구투여하였다. 경구투여 10일 종료 후, 1일째에 마우스 당 각 인플루엔자 항체 1㎍의 독감 백신을 근육 내 주사하였다. 백신 접종 후 4주째에 부검을 통해서 혈청 및 비장세포를 분리하였다.In the experimental animal of the present invention, a Balb/c 8-week-old female mouse was purchased from Orient Bio, and the cyclophosphamide (Sigma) 150mg/kg was injected intraperitoneally three times at intervals of 2 days. The experimental group was orally administered 500mg/kg of health extract for 10 days from the 3rd day after the last anticancer drug administration, and the control group was orally administered distilled water. After the end of the oral administration on the 10th day, on the first day, a flu vaccine of 1 µg of each influenza antibody per mouse was injected intramuscularly. Serum and splenocytes were isolated through autopsy at 4 weeks after vaccination.

상기 백신은 2017-2018 seasonal vaccine으로, A/Michigan/45/2015(H1N1), A/Hong Kong/4801/2014(H3N2), B/Phuket/3073/2013, B/Brisbane/60/2008을 포함하는 것이다.The vaccine is a 2017-2018 seasonal vaccine, including A/Michigan/45/2015(H1N1), A/Hong Kong/4801/2014(H3N2), B/Phuket/3073/2013, B/Brisbane/60/2008 It is to do.

실시예Example 2. ELISA를 이용한 항원 특이 항체 형성 여부 확인 2. Confirmation of antigen-specific antibody formation using ELISA

4가 백신 용액을 이용하여, 각각의 인플루엔자 항원이 1㎍/㎖씩 로딩되도록 되도록 웰당 총 4㎍/㎖의 항원을 96웰 플레이트에 코팅하고 블로킹한 후, 마우스 혈청을 10,000배 희석해서 인큐베이션 하고, HRP가 붙어있는 anti-IgG 항체, anti-IgG1 항체, anti-IgG2a 항체(각각은 1,000배 희석)를 이용해서 인플루엔자 항원 특이 항체 titer를 효소결합면역흡착검사법(ELISA)으로 측정하였다.Using a tetravalent vaccine solution, a total of 4 μg/ml of antigen per well was coated on a 96-well plate and blocked so that each influenza antigen was loaded at 1 μg/ml, and then the mouse serum was diluted 10,000 times and incubated. Anti-IgG antibody with HRP, anti-IgG1 antibody, and anti-IgG2a antibody (each diluted 1,000 times) were used to measure the titer of the influenza antigen-specific antibody by enzyme-linked immunosorbent assay (ELISA).

그 결과 도 1에 개시한 바와 같이, 건강 추출물을 투여하지 않은 항암제 투여군에 비해 건강 추출물을 투여한 군에서의 IgG 및 IgG1의 생성이 증진된 것을 확인하였다.As a result, as disclosed in FIG. 1, it was confirmed that the production of IgG and IgG1 was improved in the group administered with the healthy extract compared to the group administered with the anticancer agent to which the healthy extract was not administered.

실시예Example 3. 3. in vitroin vitro 항원 자극에 대한 사이토카인 생성 Cytokine production against antigen stimulation

(1) IFN-γ 및 IL-4의 함량 변화 확인(1) Confirmation of changes in the content of IFN-γ and IL-4

각각의 개체마다 비장세포를 분리한 후, 각각의 웰에 anti-mouse CD28 Ab(1㎍/㎖, biolegend) 자극 군과 anti-mouse CD28 Ab+인플루엔자 항원(0.5㎍/㎖) 자극 군으로 나누어 자극을 주고, 배양해서 2일 후에 배양액의 상등액을 분리하였다. 배양액에서 각각의 사이토카인의 생성 값을 구하였다. 구체적으로 TH1 반응 평가를 위한 사이토카인 IFN-γ 및 TH2 반응 평가를 위한 사이토카인 IL-4의 함량 변화를 확인하였다. After separating splenocytes for each individual, stimulation was performed in each well by dividing into an anti-mouse CD28 Ab (1㎍/㎖, biolegend) stimulation group and an anti-mouse CD28 Ab + influenza antigen (0.5㎍/mL) stimulation group. Then, after 2 days of culture, the supernatant of the culture solution was separated. The production values of each cytokine were determined in the culture medium. Specifically, changes in the content of the cytokine IFN-γ for T H 1 reaction evaluation and the cytokine IL-4 for T H 2 reaction evaluation were confirmed.

그 결과 도 2에 개시한 바와 같이 IFN-γ은 항암제 투여군 및 항암제 투여 후 건강추출물을 투여한 군에서 모두 감소한 것을 확인하였고, IL-4의 함량 변화는 항암제만 투여한 군에서는 오히려 아무것도 투여하지 않은 대조군에 비해 IL-4의 함량이 증가하였으며, 항암제 투여 후, 본 발명의 건강추출물을 투여한 군에서는 아무것도 투여하지 않은 대조군에 비해 감소하였다. As a result, as disclosed in FIG. 2, it was confirmed that IFN-γ decreased both in the group administered with the anticancer agent and the group administered with the health extract after the administration of the anticancer agent, and the change in the content of IL-4 was rather nothing in the group administered with only the anticancer agent. The content of IL-4 was increased compared to the control group, and after administration of the anticancer agent, the group to which the health extract of the present invention was administered was decreased compared to the control group to which nothing was administered.

(2) IFN-γ/ IL-4의 비율 확인(2) Confirmation of the ratio of IFN-γ/IL-4

상기 IFN-γ 및 IL-4의 함량의 변화를 확인한 후, 이들의 함량비를 분석하였다. After confirming the change in the contents of IFN-γ and IL-4, the content ratio thereof was analyzed.

그 결과, 도 3에 개시한 바와 같이 항암제만 투여했을 때 감소했던 IFN-γ/ IL-4의 비율이 항암제 및 건강 추출물을 투여한 군에서 증진된 것을 확인하였다. As a result, it was confirmed that the ratio of IFN-γ/IL-4 decreased when only the anticancer agent was administered as disclosed in FIG. 3 was improved in the group to which the anticancer agent and health extract were administered.

이후, 도 4에 개시한 바와 같이, IFN-γ/IL-4의 비율과 IgG 생성량과의 관계를 확인한 결과, 정비례관계가 있다는 것을 확인하였다.Thereafter, as disclosed in FIG. 4, as a result of confirming the relationship between the ratio of IFN-γ/IL-4 and the amount of IgG produced, it was confirmed that there is a direct proportional relationship.

실시예Example 4. 적혈구 응집억제 검사를 이용한 중화항체 4. Neutralizing antibody using red blood cell aggregation inhibition test 생성율Production rate 측정 Measure

백신을 접종하고 30일이 경과한 후, 적혈구 응집 억제 검사를 실시하였다. After 30 days of vaccination had elapsed, a hemagglutination inhibition test was performed.

그 결과, 표 1 및 2에 개시한 바와 같이, 항암제만 투여한 군에 비해 항암제 투여 후 본 발명의 건강 추출물을 투여한 군의 중화항체(항체가 40 이상의 항체) 생성율이 증진된 것을 확인하였다.As a result, as disclosed in Tables 1 and 2, it was confirmed that the production rate of neutralizing antibodies (antibodies having an antibody of 40 or more) was increased in the group to which the health extract of the present invention was administered after administration of the anticancer agent compared to the group to which only the anticancer agent was administered.

건강 추출물을 투여한 군의 H1N1에 대한 중화항체(항체가 40 이상의 항체) 생성율 The production rate of neutralizing antibodies (antibodies with an antibody of 40 or more) against H1N1 in the group administered with the healthy extract H1N1
H1N1
혈청방어(seroprotection);
적혈구 응집 억제 titer≥40
Serum protection;
Hemagglutination inhibition titer≥40
중화항체 생성율(%)
Neutralizing antibody production rate (%)
40 이상40 or more 40 미만Less than 40 항암제 단독투여Anticancer drug alone 22 66 25.025.0 항암제 투여 후, 건강 추출물 투여After administration of anticancer drug, administration of health extract 66 22 75.075.0

건강 추출물을 투여한 군의 H3N2에 대한 중화항체(항체가 40 이상의 항체) 생성율 Production rate of neutralizing antibodies (antibodies with an antibody of 40 or more) against H3N2 in the group administered with the healthy extract H3N2
H3N2
혈청방어(seroprotection);
적혈구 응집 억제 titer≥40
Serum protection;
Hemagglutination inhibition titer≥40
중화항체 생성율(%)
Neutralizing antibody production rate (%)
40 이상40 or more 40 미만Less than 40 항암제 단독투여Anticancer drug alone 1One 77 12.512.5 항암제 투여 후, 건강 추출물 투여After administration of anticancer drug, administration of health extract 22 66 25.025.0

Claims (6)

건강 추출물을 유효성분으로 포함하는 독감 백신 접종에 의해 형성되는 H1N1 또는 H3N2의 A형 독감바이러스 감염에 대한 중화항체 또는 B/Phuket 또는 B/Bristone의 B형 독감바이러스 감염에 대한 중화항체 생성율 증진용 건강기능식품 조성물.Health for enhancing the production rate of neutralizing antibodies against H1N1 or H3N2 type A influenza virus infection or B/Phuket or B/Bristone neutralizing antibody against type B influenza virus infection formed by influenza vaccination containing health extract as an active ingredient Functional food composition. 제1항에 있어서, 상기 건강 추출물의 추출 용매는 물, C1~C4의 저급 알코올 또는 이들의 혼합물인 것을 특징으로 하는 독감 백신 접종에 의해 형성되는 H1N1 또는 H3N2의 A형 독감바이러스 감염에 대한 중화항체 또는 B/Phuket 또는 B/Bristone의 B형 독감바이러스 감염에 대한 중화항체 생성율 증진용 건강기능식품 조성물.The method of claim 1, wherein the extraction solvent of the healthy extract is water, a lower alcohol of C 1 to C 4 or a mixture thereof. A health functional food composition for enhancing the production rate of neutralizing antibodies or neutralizing antibodies against B/Phuket or B/Bristone influenza virus infection. 삭제delete 제1항에 있어서, 상기 조성물은 면역력이 저하된 암환자 또는 60세 이상의 노인에게 독감 백신 접종 전에 투여하기 위한 것임을 특징으로 하는 H1N1 또는 H3N2의 A형 독감바이러스 감염에 대한 중화항체 또는 B/Phuket 또는 B/Bristone의 B형 독감바이러스 감염에 대한 중화항체 생성율 증진용 건강기능식품 조성물.The method of claim 1, wherein the composition is a neutralizing antibody against H1N1 or H3N2 type A flu virus infection or B/Phuket, which is for administration to a cancer patient with reduced immunity or an elderly person aged 60 years or older prior to inoculation with the flu. Health functional food composition for enhancing the production rate of neutralizing antibodies against B/Bristone's B-type flu virus infection. 건강 추출물을 유효성분으로 포함하는 독감 백신 접종에 의해 형성되는 H1N1 또는 H3N2의 A형 독감바이러스 감염에 대한 중화항체 또는 B/Phuket 또는 B/Bristone의 B형 독감바이러스 감염에 대한 중화항체 생성율 증진용 약학 조성물.Pharmaceuticals for enhancing the production rate of neutralizing antibodies against H1N1 or H3N2 type A influenza virus infection or B/Phuket or B/Bristone neutralizing antibody against type B influenza virus infection formed by influenza vaccination containing health extract as an active ingredient Composition. 건강 추출물을 유효성분으로 포함하는 H1N1 또는 H3N2의 A형 독감바이러스 감염에 대한 중화항체 또는 B/Phuket 또는 B/Bristone의 B형 독감바이러스 감염에 대한 중화항체 생성율 증진용 독감 백신 보조제.A flu vaccine adjuvant for enhancing the production rate of neutralizing antibodies against H1N1 or H3N2 type A influenza virus infection, or B/Phuket or B/Bristone including a healthy extract as an active ingredient.
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