KR101869322B1 - Medicinal composition and health functional food for preventing or treating of stroke sequelae - Google Patents

Medicinal composition and health functional food for preventing or treating of stroke sequelae Download PDF

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KR101869322B1
KR101869322B1 KR1020170005172A KR20170005172A KR101869322B1 KR 101869322 B1 KR101869322 B1 KR 101869322B1 KR 1020170005172 A KR1020170005172 A KR 1020170005172A KR 20170005172 A KR20170005172 A KR 20170005172A KR 101869322 B1 KR101869322 B1 KR 101869322B1
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stroke
acid
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김동현
박혜진
장대식
류종훈
이영춘
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동아대학교 산학협력단
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Abstract

The present invention relates to a pharmaceutical composition and health functional food for preventing or treating stroke sequelae which include echinocystic acid, thereby promoting the neurite outgrowth of nerve cells and exhibiting the drug efficacy and health functions for stroke sequelae in which a nerve injury occurs.

Description

뇌졸중 후유증의 예방 또는 치료용 의약 조성물 및 건강기능식품{MEDICINAL COMPOSITION AND HEALTH FUNCTIONAL FOOD FOR PREVENTING OR TREATING OF STROKE SEQUELAE}TECHNICAL FIELD [0001] The present invention relates to a medicinal composition for preventing or treating stroke sequelae,

본 발명은 뇌졸중 후유증의 예방 또는 치료용 의약 조성물 및 건강기능식품에 관한 것이다.The present invention relates to a pharmaceutical composition and a health functional food for preventing or treating stroke sequelae.

뇌졸중은 뇌로 가는 혈관이 막히거나 파열되어 생기는 일시적 뇌질환으로 심각한 후유증을 남기는 질환이다. 뇌졸중으로 발생하는 뇌신경 세포 손상의 원인이 과도한 흥분성 신경전달물질의 유리, 자유라디칼의 생성， 단백질 합성의 저해， 유전자 발현 이상 및 면역반응의 활성화 등으로 설명될 수 있으나， 아직까지 뇌신경세포 손상기전의 복잡성 등으로 뇌졸중으로 발생하는 뇌신경세포의 손상을 보호해 줄 수 있는 치료제가 개발되어 있지 못한 실정이다.Stroke is a temporary brain disease caused by clogging or rupture of the blood vessels to the brain, leaving severe aftereffects. Cerebral nerve cell damage caused by stroke can be explained by excessive excitatory neurotransmitter release, free radical generation, inhibition of protein synthesis, gene expression abnormality, and activation of immune response. However, A therapeutic agent capable of protecting brain neuronal cell damage caused by a stroke due to complexity has not been developed.

뇌졸중은 운동기능의 손실, 감각 이상, 인지, 언어장애, 의식 소실, 삼킴 기능의 장애 등 출혈 및 폐쇄 부위에 따라 다앙한 신경학적 증상을 일으킨다. 이후 다양한 신경학적 증상은 90% 이상이 대개 3개월 이내에 회 복되는 것으로 알려져 있는데 이러한 기간 동안 재활치료를 통해 개선율을 높인다. 하지만 이러한 재활기간에 재활률을 높이기 위한 특별한 약물은 개발되어 있지 않은 실정이다.Stroke causes a variety of neurological symptoms, depending on the site of bleeding and obstruction, such as loss of motor function, sensory abnormalities, cognition, speech disorders, loss of consciousness, impairment of swallowing function. Afterwards, various neurological symptoms are known to recover within 90% of cases, usually within 3 months. During this period, rehabilitation improves the recovery rate. However, no specific drug has been developed to increase the rehabilitation rate during this rehabilitation period.

신경돌기성장은 뉴런의 분화의 표지로 뉴런 간의 기능적인 네트워크 형성에 중요하며 이러한 신경돌기성장의 조절은 고령화에 따른 다앙한 신경질환을 치료할 수 있는 타깃이 될 수 있다. 에키노시스트산은 penta cyclic tri terpenoid로 항염증 및 항암 효과뿐만 아니라 항HIV 효과， 항진균 효과와 더불어 심혈관 보호 효과가 있다고 보고되어 있다. 신경계에 대한 효과로 tetradecanoylphorbol-13-acetate에 의한 치매 증상을 개선할 수 있다는 연구결과가 있으나 아직 뇌졸중에 대한 연구는 이루어 지지 않았다.Neurite outgrowth is a marker of neuronal differentiation and is important for the formation of a functional network between neurons, and the control of these neurite outgrowths can be a target for the treatment of multiple neuronal diseases due to aging. Echinosist acid is a penta cyclic tri-terpenoid, which has anti-inflammatory and anticancer effects as well as anti-HIV and anti-fungal effects as well as cardiovascular protection. Studies have shown that tetradecanoylphorbol-13-acetate can improve the symptoms of dementia by its effects on the nervous system, but no studies have been done on stroke yet.

한국공개특허 제10-2009-0049171호Korean Patent Laid-Open No. 10-2009-0049171

본 발명은 신경세포 신경돌기 성장 촉진 효과를 나타내어 우수한 약효를 갖는 뇌졸중 후유증의 예방 또는 치료용 의약 조성물을 제공하는 것을 목적으로 한다.Disclosure of Invention Technical Problem [8] The present invention provides a medicinal composition for preventing or treating cerebral apoplexy, which exhibits neuronal neurite outgrowth-promoting effect and has excellent pharmaceutical efficacy.

본 발명은 신경세포 신경돌기 성장 촉진 효과를 나타내어 우수한 기능성을 갖는 건강기능식품을 제공하는 것을 목적으로 한다.It is an object of the present invention to provide a health functional food exhibiting an effect of promoting neuronal cell neurite outgrowth and having excellent functionality.

1. 에키노시스트산(echinocystic acid)을 포함하는 뇌졸중 후유증 예방 또는 치료용 의약 조성물.1. A pharmaceutical composition for preventing or treating stroke sequelae comprising echinocystic acid.

2. 위 1에 있어서, 상기 에키노시스트산은 한련초에서 분리된 것인 뇌졸중 후유증 예방 또는 치료용 의약 조성물.2. The pharmaceutical composition for prevention or treatment of stroke sequelae according to 1 above, wherein the echinocysteic acid is isolated from Hansikucho.

3. 위 1에 있어서, 상기 뇌졸중 후유증은 운동 기능 손실, 감각 이상, 인지 장애, 언어 장애, 의식 소실 및 삼킴 기능 장애로 이루어진 군에서 선택된 것인 뇌졸중 후유증 예방 또는 치료용 의약 조성물.3. The pharmaceutical composition for prevention or treatment of stroke sequelae according to 1 above, wherein the stroke sequelae are selected from the group consisting of loss of motor function, sensory disorders, cognitive disorders, speech disorders, loss of consciousness, and swallowing dysfunction.

4. 에키노시스트산(echinocystic acid)을 포함하는 뇌졸중 후유증 예방 또는 개선용 건강기능식품.4. A health functional food for preventing or improving stroke sequelae including echinocystic acid.

5. 위 4에 있어서, 상기 에키노시스트산은 한련초에서 분리된 것인 뇌졸중 후유증 예방 또는 개선용 건강기능식품.5. The health functional food according to item 4 above, wherein the echinocysteic acid is isolated from Hansikucho.

6. 위 4에 있어서, 상기 뇌졸중 후유증은 운동 기능 손실, 감각 이상, 인지 장애, 언어 장애, 의식 소실 및 삼킴 기능 장애로 이루어진 군에서 선택된 것인 뇌졸중 후유증 예방 또는 개선용 건강기능식품.6. The health functional food according to item 4 above, wherein the stroke sequelae are selected from the group consisting of a loss of motor function, a sensory disorder, a cognitive disorder, a speech disorder, a loss of consciousness and a dysfunction of swallowing.

본 발명의 조성물은 신경세포의 신경돌기성장을 촉진하여 신경 손상이 나타나는 뇌졸증 후유증에 대한 우수한 예방, 치료 효과를 나타낼 수 있다. 이에, 뇌졸중 후유증에 따른 재활을 돕는 의약, 건강기능식품으로서 우수하게 활용될 수 있다.The composition of the present invention can exhibit an excellent preventive and therapeutic effect on stroke sequelae which promote neurite outgrowth of nerve cells and cause nerve damage. Therefore, it can be used as a medicine and a health functional food which help rehabilitation after a stroke after stroke.

도 1은 신경돌기성장에 미치는 에키노시스트산의 효과를 나타낸 것이다. (A) 각 군의 신경돌기성장을 평가하기 위해 사용한 대표 사진으로, RA는 양성대조군으로 사용한 retinoic acid를 말한다. (B) 신경돌기의 성장을 보인 세포의 비율을 나타낸 막대그래프로, 에키노시스트산은 농도 의존적으로 신경돌기성장을 촉진하는 효과가 있음을 알 수 있다.
도 2는 중뇌동맥 결찰 및 재관류 후 신경 손상 정도 평가 결과를 나타낸 것이다. EA는 에키노시스트산을 지칭한다. 재관류 후 하루 한 번씩 neurological deficit score를 측정하였다. 7일째 되는 날부터 1일 1회 EA(5 mg/kg)을 경구투여하였다. 재관류 후 11일(EA 투여 후 5일째 되는 날부터 약물을 먹이지 않은 그룹과 유의적인 score 차이가 남을 알 수 있다.Figure 1 shows the effect of echinocysteic acid on neurite outgrowth. (A) Representative photographs used to evaluate neurite outgrowth of each group. RA refers to retinoic acid used as a positive control. (B) A bar graph showing the percentage of cells showing the growth of neurites. It can be seen that the echinocysteic acid has an effect of stimulating neurite growth in a concentration-dependent manner.
FIG. 2 shows the results of evaluating the degree of neuronal damage after middle cerebral artery ligation and reperfusion. EA refers to an echinocysteic acid. The neurological deficit score was measured once a day after reperfusion. From day 7, EA (5 mg / kg) was orally administered once a day. After reperfusion, it was found that there was a significant difference in score between day 11 and day 5 after EA administration.

이하 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.

본 발명의 뇌졸중 후유증 예방 또는 치료용 의약 조성물은 에키노시스트산(echinocystic acid)을 포함한다.The pharmaceutical composition for preventing or treating stroke sequelae of the present invention includes echinocystic acid.

에키노시스트산은 하기 화학식 1로 표시되는 구조를 갖는 화합물로서, 항염증, 항암 효과 등이 알려져 있으나, 뇌졸중과 관련된 효과는 기존에 알려져 있지 않다.The echinocysteic acid has a structure represented by the following formula (1), and its antiinflammatory and anticancer effects are known, but the effects related to stroke have not been known.

[화학식 1][Chemical Formula 1]

Figure 112017003924189-pat00001
.
Figure 112017003924189-pat00001
.

본 명세서에서 뇌졸중 후유증(Stroke sequelae)은 뇌졸중을 겪은 이후에 발생하는 후유증을 의미하는 것으로, 구체적으로 운동 기능 손실, 감각 이상, 인지 장애, 언어 장애, 의식 소실, 삼킴 기능 장애 등을 의미한다.Stroke sequelae in the present specification means sequelae occurring after suffering a stroke, and specifically refers to loss of motor function, sensory abnormality, cognitive disorder, speech disorder, loss of consciousness, swallowing dysfunction, and the like.

본 발명자들은 상기 에키노시스트산이 신경돌기 성장 촉진 효과를 나타내어 우수한 신경재생, 보완 효과가 있어, 뇌졸중 후유증을 개선, 치료할 수 있음에 착안하여 본 발명을 고안하였다.The present inventors have devised the present invention in consideration of the fact that the above-mentioned echinocysteic acid exhibits an effect of promoting neurite outgrowth and has excellent nerve regeneration and complementary effect, thereby improving and treating stroke sequelae.

본 발명에 따른 에키노시스트산은 예를 들면 한련초에서 분리된 것일 수 있고, 또한 시그마 알드리치사(Sigma-Aldrich) 등에서 판매되는 것일 수도 있으나, 이에 제한되는 것은 아니다.The echinocysteic acid according to the present invention may be, for example, isolated from Hansucikcho, or may be from Sigma-Aldrich, but is not limited thereto.

본 발명의 의약 조성물은 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화하여 사용될 수 있다.The pharmaceutical composition of the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions according to a conventional method.

본 발명의 추출물을 함유하는 조성물에 함유될 수 있는 담체, 부형제 및 희석제로는 락토오즈(lactose), 덱스트로즈, 수크로스(sucrose), 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 및 광물유를 들 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다.Examples of carriers, excipients and diluents that may be contained in the composition containing the extract of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, , Alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil have. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used.

경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 상기 화합물에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘카보네이트(calcium carbonate), 수크로스 또는 락토오스, 젤라틴 등을 섞어 조제된다. 또한 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다.Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose or lactose , Gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included .

비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제가 포함된다. 비수성용제, 현탁제로는 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.Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

본 발명의 의약 조성물의 사용량은 환자의 나이, 성별, 체중에 따라 달라질 수 있으나, 0.1 내지 100mg/kg으로, 바람직하게는 1 내지 10mg/kg을 일일 1회 내지 수회 투여할 수 있다. 또한 그 투여량은 투여경로, 질병의 정도, 성별, 체중, 나이 등에 따라서 증감될 수 있다. 따라서 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다.The dosage of the pharmaceutical composition of the present invention may vary depending on the age, sex and body weight of the patient, but may be 0.1 to 100 mg / kg, preferably 1 to 10 mg / kg, once to several times per day. The dosage may also be increased or decreased depending on the route of administration, degree of disease, sex, weight, age, and the like. Accordingly, the dosage is not limited in any way to the scope of the present invention.

또한, 본 발명은 뇌졸중 후유증 예방 또는 개선용 건강기능식품을 제공한다.In addition, the present invention provides a health functional food for preventing or improving stroke sequelae.

본 발명의 뇌졸중 후유증 예방 또는 개선용 건강기능식품은 에키노시스트산(echinocystic acid)을 포함한다.The health functional food for preventing or improving stroke sequelae of the present invention includes echinocystic acid.

본 발명의 건강기능식품은 상기 추출물을 포함한 음료 (알콜성 음료 포함), 과실 및 그의 가공식품 (예: 과일통조림, 병조림, 잼, 마아말레이드 등), 어류, 육류 및 그 가공식품 (예: 햄, 소시지콘비이프 등), 빵류 및 면류 (예: 우동, 메밀국수, 라면, 스파게티, 마카로니 등), 과즙, 각종 드링크, 쿠키, 엿, 유제품 (예: 버터, 치즈 등), 식용식물유지, 마아가린, 식물성 단백질, 레토르트 식품, 냉동식품, 각종 조미료(예: 된장, 간장, 소스 등) 등일 수 있다. The health functional food of the present invention can be used as a health functional food containing the above extract (including an alcoholic beverage), fruit and its processed food (e.g., canned fruit, jar, jam, maralade, etc.), fish, (Eg udon, buckwheat noodles, ramen noodles, spaghetti, macaroni, etc.), juice, various drinks, cookies, candy, dairy products such as butter, cheese, Margarine, vegetable protein, retort food, frozen food, various seasonings (eg, miso, soy sauce, sauces, etc.).

또한, 본 발명의 건강기능식품은 정제, 환제, 산제, 과립제, 분말제, 캡슐제, 액제 제형 등으로 제형화된 것일 수도 있다. 이들은 담체, 희석제, 부형제 및 첨가제 중 하나 이상을 더 포함하여 제형화될 수 있다.In addition, the health functional food of the present invention may be formulated into tablets, pills, powders, granules, powders, capsules, liquid preparations and the like. These may be formulated further comprising one or more of carriers, diluents, excipients and additives.

본 발명에 더 포함될 수 있는 첨가제로는, 천연 탄수화물, 향미제, 영양제, 비타민, 광물(전해질), 풍미제(합성 풍미제, 천연 풍미제 등), 착색제, 충진제(치즈, 초콜렛 등), 팩트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH조절제, 안정화제, 방부제, 산화 방지제, 글리세린, 알콜, 탄산화제 및 과육으로 이루어진 군으로부터 선택된 1종 이상의 성분을 사용할 수 있다. Examples of the additive which can be further included in the present invention include natural carbohydrates, flavors, nutrients, vitamins, minerals (electrolytes), flavors (synthetic flavors, natural flavors and the like), colorants, fillers At least one component selected from the group consisting of acids and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, antioxidants, glycerin, alcohols, carbonating agents and fats can be used.

상술한 천연 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토스, 슈크로스 등; 및 폴리사카라이드, 예를 들어 덱스트린, 시클로덱스트린 등과 같은 통상적인 당, 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 상기 향미제로서 천연 향미제(타우마틴, 스테비아 추출물(예를 들어 레바우디오시드 A, 글리시르히진등) 및 합성 향미제(사카린, 아스파르탐 등)를 유리하게 사용할 수 있다. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavors (tautatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharine, aspartame, etc.) can be advantageously used as the flavor.

상기 외에 본 발명의 건강기능식품은 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 중진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있다. 그 밖에 본 발명의 건강기능식품은 천연 과일 쥬스 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. In addition to the above, the health functional food of the present invention may contain flavorings such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and heavies (cheese, chocolate etc.), pectic acid and its salts, And salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the health functional food of the present invention may contain flesh for the production of natural fruit juice and vegetable beverages. These components may be used independently or in combination.

상기 담체, 부형제, 희석제 및 첨가제의 구체적인 예로는 이에 한정하는 것은 아니나, 락토즈, 덱스트로즈, 슈크로즈, 솔비톨, 만니톨, 에리스리톨, 전분, 아카시아 고무, 인산칼슘, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 미세결정성 셀룰로즈, 폴리비닐피롤리돈, 셀룰로즈, 폴리비닐피롤리돈, 메틸셀룰로즈, 물, 설탕시럽, 메틸셀룰로즈, 메틸 하이드록시 벤조에이트, 프로필하이드록시 벤조에이트, 활석, 스테아트산 마그네슘 및 미네랄 오일로 이루어진 그룹으로부터 선택된 1종 이상이 사용되는 것이 바람직하다.Specific examples of the carrier, excipient, diluent and additives include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, erythritol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium phosphate, calcium Silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, polyvinylpyrrolidone, methylcellulose, water, sugar syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate And a mineral oil are preferably used.

상기 상술한 제형 내 유효성분으로서의 본 발명의 건강기능식품의 함량은 사용 형태 및 목적, 사용자 상태, 증상의 종류 및 경중 등에 의하여 적절하게 조절할 수 있으며, 고형분 중량 기준으로 0.001 내지 99.9 중량%, 바람직하게는 0.01 내지 50 중량%일 수 있으나, 이에 한정되지 않는다.The content of the health functional food of the present invention as the active ingredient in the above-described formulation can be appropriately adjusted depending on the mode and purpose of use, the condition of the user, the type of symptoms and the severity, and is 0.001 to 99.9% by weight, May be 0.01 to 50% by weight, but is not limited thereto.

이하, 본 발명을 구체적으로 설명하기 위해 실시예를 들어 상세하게 설명하기로 한다. BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to examples.

실시예Example

1. 약물 및 시약1. Drugs and reagents

에키노시스트 산은 최초 한련초(경동시장에서 구입, Seoul, Korea)에서 분리하여 사용하였고, 이후 Sigma-Aldrich Chemistry Co.에서 구입하여 사용하였다.Echinosist acid was first used in Hansikschu (purchased from Kyungdong market, Seoul, Korea) and then purchased from Sigma-Aldrich Chemistry Co.

2. 실험동물의 준비2. Preparation of experimental animals

6주령의 ICR 마우스 (26~28g)를 (주)샘타코에서 공급받아 동아대학교 생명자원과학대학의 clean cage에 약 5일간 적응시켜 사용하였으며, 물과 사료는 자유롭게 섭취하도록 하였고， 온도(23±2℃). 습도 (55±10%) 및 명암주기 (12 시간)는 자동으로 조절되도록 하였다.Six - week - old ICR mice (26 ~ 28g) were supplied from Sam Taco Co., Ltd. and used for 5 days in a clean cage of Dong - A University College of Life Science and Technology. Water and feed were freely consumed. 2 [deg.] C). The humidity (55 ± 10%) and the light period (12 hours) were adjusted automatically.

3. 통계처리3. Statistical processing

모든 실험 결과는 ANOVA (one way analysis of variance)를 이 용하여 통계처리하였고, 유의성이 인정될 경우 Student-Newman-Keuls Test를 사용하여 p<0.05 수준 이하에서 유의성 검정을 실시하였다.Statistical analysis was performed using ANOVA (one way analysis of variance). Statistical significance was tested at the p <0.05 level using the Student-Newman-Keuls test when significance was recognized.

실험예Experimental Example 1. 신경돌기성장 평가 1. Neurite outgrowth assessment

(1) 실험 방법(1) Experimental method

신경돌기성장을 측정하기 위해서 Neuro2a 세포를 24-we11 cu1ture p1ate(2 x 104cells/well)에 24시간 배앙 후， FBS가 1~2% 포함된 Dublecco’s Modified Eag1e's Medium(Welgene, korea)에 Retinoic acid 15μM， Echinocystic acid를 각각 5μM. 10μM, 15μM, 20μM 농도로 희석한 후 세포에 6시간 동안 처리하였다. 그 후， 위상차 현미경을 200x 배율로 하여 관찰하였다. Neurite-bearing ce11은 신경돌기의 길이가 세포체의 지름보다 긴 신경돌기를 가진 세포로 정의하였으며 무작위로 3~5곳을 선택하여 측정하였다.To measure neurite outgrowth, Neuro2a cells were cultured in 24-we11 cu1ture p1ate (2 x 10 4 cells / well) for 24 hours, and then Dublecco's Modified Eagle's Medium (Welgene, Korea) containing 1-2% 15 μM and Echinocystic acid, respectively. 10 [mu] M, 15 [mu] M and 20 [mu] M and then treated for 6 hours. Thereafter, the phase difference microscope was observed at a magnification of 200x. Neurite-bearing ce11 was defined as a cell with a neurite longer than the diameter of the cell body, and randomly selected from 3 to 5 sites.

(2) 실험 결과(2) Experimental results

앙성대조군으로 사용한 retinoic acid는 대조군에 비해 유의적으로 신경돌기성장을 보인 세포 수를 증가시켰다(도 1A. 1B). 또한 echinocystic acid의 경우 농도 의존적으로 신경돌기성장을 보인 세포 수를 증가시켰다(도 1A, 1B).The retinoic acid used as a control group increased the number of cells showing neurite outgrowth as compared with the control group (Fig. 1A. 1B). In addition, echinocystic acid increased the number of cells showing neurite outgrowth in a concentration-dependent manner (Fig. 1A, 1B).

실험예Experimental Example 2. 뇌졸중 후유증 평가 2. Evaluation of stroke sequelae

(1) 실험방법(1) Experimental method

중뇌동맥 결찰모델을 이용하여 에키노시스트산의 효과를 확인 하였다. 중뇌동맥 결찰 모델에 나일론 팔라멘트를 내경동맥(internal carotid artery)으로 삽입하여 중뇌동맥을 폐색시킨 후 120분 후에 다시 필라멘트를 제거함으로써 재관류시키는 방법이다. 에키노시스트산이 뇌졸중 후유증으로 인한 신체기능장애에 대한 재활을 돕는지 평가하기 위해 중뇌동맥 결찰 및 재관류 1주일 후 에키노시스트산을 5mg/kg의 농도로 매일 투여 하며 1주일간 신경학적 기능(neurological score) 평가를 실시하였다.The effect of echinocysteic acid was confirmed using the middle cerebral artery ligation model. In the middle cerebral artery ligation model, a nylon palate is inserted into the internal carotid artery to occlude the middle cerebral artery, and after 120 minutes, the filament is removed again to reperfusion. To evaluate whether echinosistan acid helps rehabilitate the dysfunction due to sequelae of stroke, echinocysteic acid is administered daily at a concentration of 5 mg / kg once a week after ligation of the middle cerebral artery and reperfusion, and neurological score ).

(2) 실험 결과(2) Experimental results

도 2를 참조하면, 재관류 후 11일(EA 투여 후 5일째 되는 날부터 약물을 먹이지 않은 그룹과 유의적인 score 차이가 나타나, 신경학적 기능의 변화가 유의적으로 차이가 남을 알 수 있다.Referring to FIG. 2, there is a significant difference in score between the drug-free group and the change in neurological function from day 5 after EA administration to 11 days after reperfusion.

Claims (6)

에키노시스트산(echinocystic acid)을 포함하는 뇌졸중에 따른 운동 기능 손실 예방 또는 치료용 의약 조성물.
A pharmaceutical composition for preventing or treating loss of motor function due to stroke including echinocystic acid.
청구항 1에 있어서, 상기 에키노시스트산은 한련초에서 분리된 것인 뇌졸중에 따른 운동 기능 손실 예방 또는 치료용 의약 조성물.
[Claim 2] The pharmaceutical composition according to claim 1, wherein the echinocystein is isolated from the muscles of a mammal.
삭제delete 에키노시스트산(echinocystic acid)을 포함하는 뇌졸중에 따른 운동 기능 손실 예방 또는 개선용 건강기능식품.
A health functional food for preventing or ameliorating loss of exercise function due to stroke including echinocystic acid.
청구항 4에 있어서, 상기 에키노시스트산은 한련초에서 분리된 것인 뇌졸중에 따른 운동 기능 손실 예방 또는 개선용 건강기능식품.[Claim 5] The health functional food according to claim 4, wherein the echinocystein is isolated from the genus Hancock. 삭제delete
KR1020170005172A 2017-01-12 2017-01-12 Medicinal composition and health functional food for preventing or treating of stroke sequelae KR101869322B1 (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109549942A (en) * 2019-01-16 2019-04-02 于海龙 Application of the echinocystic acid in treatment cerebral infarction medicine preparation
KR20240032613A (en) 2022-08-30 2024-03-12 서울대학교산학협력단 The pharmaceutical composition for relivieing or treating affereffects of cerebral infarction

Citations (2)

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Publication number Priority date Publication date Assignee Title
KR20090049171A (en) 2007-11-13 2009-05-18 관동대학교산학협력단 Preventing and modulating agents of cerebral apoplexy using vasodilatation effects of the extract of sargassum siliquastrum or its effective constituent
KR20140077382A (en) * 2012-12-14 2014-06-24 경희대학교 산학협력단 Pharmaceutical Composition comprising lancemaside A or metabolite thereof for prevention and treatment of cognitive disorder

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20090049171A (en) 2007-11-13 2009-05-18 관동대학교산학협력단 Preventing and modulating agents of cerebral apoplexy using vasodilatation effects of the extract of sargassum siliquastrum or its effective constituent
KR20140077382A (en) * 2012-12-14 2014-06-24 경희대학교 산학협력단 Pharmaceutical Composition comprising lancemaside A or metabolite thereof for prevention and treatment of cognitive disorder

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109549942A (en) * 2019-01-16 2019-04-02 于海龙 Application of the echinocystic acid in treatment cerebral infarction medicine preparation
KR20240032613A (en) 2022-08-30 2024-03-12 서울대학교산학협력단 The pharmaceutical composition for relivieing or treating affereffects of cerebral infarction

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