WO2015072596A1 - Method for preparing medicinal herb extract using malt enzyme liquid - Google Patents

Method for preparing medicinal herb extract using malt enzyme liquid Download PDF

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WO2015072596A1
WO2015072596A1 PCT/KR2013/010338 KR2013010338W WO2015072596A1 WO 2015072596 A1 WO2015072596 A1 WO 2015072596A1 KR 2013010338 W KR2013010338 W KR 2013010338W WO 2015072596 A1 WO2015072596 A1 WO 2015072596A1
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extract
group
malt enzyme
cisplatin
malt
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PCT/KR2013/010338
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French (fr)
Korean (ko)
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박영준
여말희
김동규
이송진
김세환
유재훈
박재홍
강지윤
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씨제이헬스케어 주식회사
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Priority to PCT/KR2013/010338 priority Critical patent/WO2015072596A1/en
Publication of WO2015072596A1 publication Critical patent/WO2015072596A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
    • A61K36/489—Sophora, e.g. necklacepod or mamani
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/899—Poaceae or Gramineae (Grass family), e.g. bamboo, corn or sugar cane
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine

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  • the present invention relates to a method for producing a herbal extract using malt enzyme liquid, and more specifically, reacting the malt enzyme liquid and the herbal medicine; And it relates to a method for producing a herbal extract having an appetite and immune enhancing effect prepared by the step of extracting the reacted malt enzyme solution and herbal medicine.
  • Anorexia nervosa (or anorexia nervosa, anorexia) can cause psychotic symptoms such as characteristic cravings for emaciation and abnormal eating behavior, as well as body weight loss and amenorrhea (for women) of 20% or more of standard weight.
  • a symptomatic disease which often occurs in young women and is a serious and sometimes fatal disease.
  • various preparations for promoting appetite and digestion are often used.
  • Representative fermentation agents containing a large amount of digestive enzymes are widely used.
  • the core material is a yeast containing a large amount of digestive enzymes such as amylase, peptidase, and the like, followed by drying and tableting.
  • vitamin preparations, various digestive enzyme preparations or various formulations that stimulate the appetite center are used.
  • these drugs have a direct effect upon administration, but simply ingesting the appetite without the activation of other organs by simply digesting or stimulating the appetite causes the problem of worsening the disease. .
  • the present inventors while studying natural products having an effective appetite enhancing or immune enhancing activity, the long-term toxicity as a result of administering the malt enzyme extract herbal drug extract extracted by reacting the malt enzyme solution and the herbal medicine to the appetite suppression or immunosuppression animal model
  • the present invention was completed by confirming to alleviate the effects and at the same time exhibit an appetite enhancing or immune enhancing effect.
  • the present invention comprises the steps of reacting the malt enzyme solution and the herbal medicine (step 1); And extracting the reacted malt enzyme solution and the herbal medicine (step 2).
  • Step 1 is a step of reacting the malt enzyme solution with the herbal medicine to lower the molecular weight by hydrolyzing the carbohydrate and protein components of the herbal medicine.
  • the mixing ratio of the malt enzyme solution and the herbal medicine is preferably 50-99.9% by weight: 0.1-50% by weight. If the mixing ratio of the herbal medicine exceeds 50% by weight, a problem may occur that mixing is not easy.
  • the mixing is preferably mixed by stirring for 1 to 12 hours while maintaining a temperature range of 30 °C to 70 °C.
  • malt enzyme liquid used in the present invention means a liquid from which the enzyme is extracted from the malt sprouted from the barley called dried barley.
  • the malt enzyme solution of the present invention is preferably prepared by adding malt to water in an amount of 0.1% to 99% by weight and then mixing the mixture for 0.5 hours to 24 hours.
  • the reaction temperature is preferably 20 ° C to 50 ° C, more preferably 20 ° C to 40 ° C. If the reaction proceeds at a temperature range higher than 50 ° C., the enzyme activity of the malt enzyme solution may be remarkably reduced, and the extraction yield of the malt enzyme liquid may drop at a temperature range lower than 20 ° C. In view of such a problem, the reaction temperature is most preferably room temperature (about 24 ° C).
  • Step 2 is a step for obtaining the herbal extract by extracting the mixed malt enzyme solution and the herbal medicine.
  • the herbal medicine is preferably papillary or dermis.
  • extract means a specific component extracted from the mixed malt enzyme liquid and the crude drug using a solvent of the liquid.
  • a solvent of the liquid water, C 1-4 alcohol or a mixture thereof may be used.
  • concentration of ethanol is preferably 10% to 95%, but is not limited thereto.
  • the extraction may be performed for 1 to 24 hours, preferably 1 to 5 hours.
  • the extraction may use any one method selected from the group consisting of reflux cooling extraction, cold needle extraction, hot water extraction and ultrasonic extraction, it is preferable to use a reflux cooling extraction method in order to efficiently extract the active ingredient.
  • the extract may be obtained in a liquid or powder form by concentrating under reduced pressure or drying under reduced pressure in the temperature range of 20 °C to 100 °C as necessary, but is not limited thereto.
  • the herbal extract method using the malt enzyme liquid according to the present invention has the effect of increasing the bioabsorption rate by hydrolyzing the carbohydrate and protein components of the herbal medicine to lower the molecular weight.
  • FIG. 1 is a graph showing the appetite-promoting effect of malt enzyme liquid wild boar extract and wild boar extract in cisplatin-induced acute anorexia animal model according to an embodiment of the present invention.
  • FIG. 2 is a graph showing the appetite-stimulating effect of the dermis extract, malt enzyme dermis extract and megestrol acetate (control drug) in cisplatin-induced acute anorexia animal model according to an embodiment of the present invention.
  • Acute appetite decay animal model was used after a 7-week-old male rats after a period of 1 week of acclimatization (8-week-old male rats, 250 g to 270 g). A total of 7 rats were divided into 8 groups (normal group, cisplatin group, and mesgentral group 6) and fasted for 24 hours under free water intake.
  • the three groups were diluted twice (bid: twice a day) megestrol acetate in 0.5% methylcellulose (MC) after 18 hours of fasting and 23.5 hours of fasting in rats 25 mg / kg, 35 mg / kg and 45 mg / Suspension was administered orally at a dose of kg, and another three groups were diluted once a day (qd: once a day) megestrol acetate in 0.5% methylcellulose (MC) 23.5 hours after fasting 50 mg / kg, 70 rats, respectively Suspension was administered orally at the dose of mg / kg and 90 mg / kg.
  • cisplatin was dissolved in 5 ml of water for injection at 2 mg / kg of rat weight and rats were injected intraperitoneally. Immediately after administration of cisplatin, one rat was placed in a cage and water was supplied freely, and 50 g of food was recorded and recorded. After 24 hours, the remaining amount of food was measured to calculate the food intake of the rats, and the difference in food intake with each group was compared. The results are shown in Table 1 below.
  • Acute anorexia animals purchased 7 week old male rats and used 250 g to 270 g 8 week old male rats that had undergone one week of acclimation. Each group was divided into 4 groups (normal group, cisplatin-only group, malt enzyme extract group extract group, group extract group). Each group fasted 24 hours under free water intake, and the herbal extract-administered group diluted each extract with 0.5% methylcellulose (MC) twice after 19 hours and 23.5 hours after fasting (bid: 1 day). Twice) 100 mg / kg wild boar extract of Comparative Example 1 and 100 mg / kg wild boil extract of malt enzyme solution of Example 1 were orally administered to rats, respectively.
  • cisplatin was dissolved in 5 ml of water for injection at 2 mg / kg of rat weight and rats were injected intraperitoneally. Immediately after administration of cisplatin, one rat was placed in a cage, and water was freely consumed. After 24 hours, the food intake of rats was calculated by measuring the amount of food remaining, and the food intakes of the cisplatin-only group and each extract-administered group were compared. The results are shown in Table 2 and FIG. 1.
  • Acute anorexia animals purchased male rats of 7 weeks of age, followed by a 1-week purifying period, and used 250- to 270 g of 8-week-old male rats. Each group was divided into 5 groups (normal group, cisplatin alone group, megestrol acetate group, malt enzyme dermal extract group, dermal extract group). Each group fasted 24 hours under free water intake, and the herbal extract-administered group diluted each extract with 0.5% methylcellulose (MC) twice after 19 hours and 23.5 hours after fasting (bid: 1 day). Twice) 600 mg / kg dermal extract of Comparative Example 2 and malt enzyme liquid dermis extract of 600 mg / kg Example 2 were orally administered to rats, respectively.
  • MC methylcellulose
  • the megestrol acetate group was orally administered at a dose of 70 mg / kg diluted in 0.5% methylcellulose (MC) once (qd: once a day) only after 23.5 hours of fasting.
  • MC 0.5% methylcellulose
  • cisplatin was dissolved in 5 ml of water for injection at 2 mg / kg of rat weight and rats were injected intraperitoneally.
  • one rat was placed in a cage, and water was freely consumed.
  • the food intake of rats was calculated by measuring the amount of food remaining, and the food intakes of the cisplatin-only group and each extract-administered group were compared. The results are shown in Table 3 and FIG. 2.
  • the cisplatin alone group showed a significant decrease in food intake compared to the normal group, 600 mg / kg group administered the dermis extract of Comparative Example 2 14.7% appetite promoting effect Showed.
  • the group administered the malt enzyme liquid dermis extract of 600 mg / kg of Example 2 was confirmed that the appetite promoting effect of 31.6% to 46.3%.
  • the appetite stimulating agent exhibits an appetite-stimulating effect equal to or higher than that of megestrol acetate, a commercially available control drug.
  • the endocrine system side effects mitigation effect of cisplatin-induced chronic anorexia animal model using malt enzyme extract triceps extract prepared in Example 1 was analyzed.
  • the chronic anorexic animal model purchased male rats of 7 weeks old and used 250 g to 270 g of 8 week old male rats that had undergone one week of acclimation. Each group was divided into four groups (normal group, cisplatin-only group, malt-enzyme sandal group extract-administered group and control group megestrol acetate-administered group) of 7 rats, and the experiment was conducted for 7 days. From the first day to the seventh day of the experiment, the malt enzyme extract wild boiled root extract of Example 1 was diluted in 0.5% methylcellulose (MC) twice at 10 am and 5 pm (bid: twice a day) at a dose of 150 mg / kg.
  • MC methylcellulose
  • the control drug megestrol acetate
  • MC 0.5% methylcellulose
  • the rest of the groups were orally administered 0.5 ml methylcellulose (MC) at a dose of 5 ml / kg.
  • cisplatin was dissolved twice in 5 ml of water for 2 kg / kg rats intraperitoneally and injected on the 1st and 2nd day of the experiment. From the first day of the experiment, one rat was placed in a cage, and water was supplied and recorded at a predetermined amount in a state in which water was freely ingested.
  • cisplatin alone-administered cisplatin alone showed a significant decrease in weight and food intake compared to normal, and showed significant changes in liver, kidney and testes in long-term weight. It was confirmed. Not only does cisplatin administration lead to a decrease in food intake and weight loss due to anorexia, but also direct and indirect side effects on endocrine organs.
  • the group administered 150 mg / kg of malt enzyme liquid wild boar extract of Example 1 was significantly improved food intake than the cisplatin alone group, it was confirmed that the decrease in weight can be alleviated.
  • the immunosuppressive animal model used male rats of 7 weeks old and used 8-week old male rats ranging from 250 g to 270 g after one week of acclimation.
  • the rats were separated into four groups (normal group, cyclophosphanide alone group, malt enzyme solution of wild bovine muscle extract group of Example 1 and control group megestrol acetate group) of 7 rats, and the experiment was conducted for 6 days. From day 1 to day 6 of the experiment, the maltose extract wild boiled root extract group of Example 1 was carried out at 10 am and 5 pm at 150 mg / kg twice the malt enzyme boiled bovine root extract of 1 (bid: twice a day).
  • the cyclophosphamide alone group showed no change in body weight compared to the normal group, but showed a significant decrease in splenic and thymus weight, which is an immune cell-related organ. This indicates that immunity was suppressed by cyclophosphamide administration.
  • the extract administration group of Example 1 showed no change in body weight compared to the cyclophosphamide alone group, but showed an effect of alleviating weight loss of cyclophosphamide-induced immune organs in both spleen and thymus.

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Abstract

The present invention relates to a medicinal herb extract using a malt enzyme liquid and a preparation method therefor and, more specifically, to a method for preparing a medicinal herb extract having appetite and immunity increasing effects, the method comprising: reacting a malt enzyme liquid with a herbal medicine; and extracting the reacted malt enzyme liquid and herbal medicine. Accordingly, the method for preparing a medicinal herb extract using a malt enzyme liquid enables carbohydrate and protein components of the herb medicine to be hydrolyzed so as to have low molecular weights, thereby providing the effect of increasing the absorption ratio in the human body.

Description

맥아 효소액을 이용한 생약 추출물의 제조방법Preparation method of herbal extract using malt enzyme solution
본 발명은 맥아 효소액을 이용한 생약 추출물의 제조방법에 관한 것으로, 더 자세하게는 맥아 효소액 및 생약을 반응시키는 단계; 및 상기 반응된 맥아 효소액 및 생약을 추출하는 단계로 제조된 식욕 증진 및 면역 증진 효과를 갖는 생약 추출물의 제조방법에 관한 것이다.The present invention relates to a method for producing a herbal extract using malt enzyme liquid, and more specifically, reacting the malt enzyme liquid and the herbal medicine; And it relates to a method for producing a herbal extract having an appetite and immune enhancing effect prepared by the step of extracting the reacted malt enzyme solution and herbal medicine.
최근 들어 사스(SARS), 조류독감(AI), 신종플루 등 신규 유행성 질환의 빈번한 발생은 전 세계적으로 심각한 경제적 손실과 국민 건강의 위해 요인이 되고 있다. 특히, 인플루엔자의 경우 세계적으로 매년 약 10억의 인구가 감염되고 30~50만 명이 이로 인해 사망한다고 추정되고 있다. 계절 독감과 신종플루 등 바이러스 질환의 경우 백신접종과 타미플루 같은 항바이러스제의 복용이 최선의 예방과 치료방법으로 알려져 있으나, 일부 신경성 부작용 문제, 수급의 어려움 및 오남용으로 인한 내성바이러스의 출현 등이 문제시되며 또한 항바이러스에 내성을 가진 신종 인플루엔자의 발견 등으로 예방과 치료에 한계를 나타내고 있는 실정이다. In recent years, the frequent outbreaks of new pandemic diseases, such as SARS, avian influenza (AI), and swine flu, are causing serious economic losses and risks to public health worldwide. Influenza, in particular, is estimated to affect approximately one billion people worldwide each year and cause 30 to 500,000 deaths. In the case of viral diseases such as seasonal flu and swine flu, vaccination and the use of antiviral drugs such as Tamiflu are known to be the best prevention and treatment methods, but some neurological side effects, difficulty in supply and the emergence of resistant viruses due to misuse, etc. In addition, due to the discovery of a new influenza resistant to antiviral, the situation is showing a limit in the prevention and treatment.
또한, 산업화와 환경의 변화로 인하여 다양한 원인에 의한 면역력 저하로 인해 각종 질병에 노출되어 있으며, 이러한 질병 치료를 위해 막대한 비용을 소모하고 있다. 특히, 최근 오염된 공기 및 식생활 변화, 즉 인스턴트 식품, 탄산음료 등의 과잉섭취 등으로 인해 면역체계에 이상이 발생하여 알러지성 질환들이 지속적으로 증가되고 있는 추세이다. In addition, due to industrialization and changes in the environment, due to various factors, immunity is lowered, and is exposed to various diseases, and consumes enormous costs for treating such diseases. In particular, allergic diseases are continuously increasing due to an abnormality in the immune system due to the recent contaminated air and dietary changes, that is, excessive intake of instant food, carbonated beverages, and the like.
이에, 알러지성 질환을 포함하는 면역질환을 예방 및 치료하기 위해, 여러 가지 약제가 개발되고 있으나, 대부분의 약제들이 화학약품들로 부작용이 있으며, 지속적인 치료효과를 내지 못하는 한계가 있다. Thus, in order to prevent and treat immune diseases including allergic diseases, various drugs have been developed, but most drugs have side effects with chemicals, and there is a limit that does not produce a continuous therapeutic effect.
따라서, 외부 세균이나 바이러스로부터 신체를 보호할 수 있는 면역력 증진 건강식품에 대한 소비자의 관심이 높아지고 있다. 이에, 마늘, 인삼 등 면역력 증진 및 개선 효과가 있는 건강기능식품 시장이 높은 성장세를 나타내고 있으나, 면역력 증진 효능과 관련하여 동물모델이나 임상연구를 통한 과학적인 효능 입증이나 신규 면역증진 건강기능 소재의 발굴 및 상품화 등 관련연구는 아직 부족한 실정이다. Therefore, consumers' interest in immune-enhancing health food that can protect the body from external bacteria and viruses is increasing. In this regard, the market for health functional foods with immunity enhancement and improvement effects, such as garlic and ginseng, is showing high growth.However, with respect to the efficacy of immunity enhancement, it has been proved scientifically through animal models or clinical studies or discovery of new immune-enhancing health functional materials. Related research such as and commercialization is still insufficient.
한편, 노화, 만성 질환 환자 및 급성 상태나 질병을 앓는 환자는 질병의 상태나 치료에 기인한 식욕부진이 발생하는데, 식욕부진은 체중 유지 곤란, 특히 몸무게 감소로 특징지어지는 영양 및 대사 감퇴를 수반하게 된다. 이에, 많은 식욕 부진을 겪는 사람들이 식사장애 또는 영양장애를 앓게 된다. 예를 들어, 암 화학요법을 받는 환자, AIDS 등에 기인한 면역타협환자, 또는 기관 이식 등에 기인한 면역억제 환자 등이 공통적으로 식욕 감퇴를 경험하며, 수술 후의 환자도 식욕 감퇴를 경험할 수 있다. On the other hand, aging, chronic disease patients and patients with acute conditions or diseases develop anorexia due to the condition or treatment of the disease, which is accompanied by nutrition and metabolic decline characterized by difficulty in maintaining weight, especially weight loss. Done. As a result, many people suffering from anorexia suffer from eating disorders or malnutrition. For example, patients undergoing cancer chemotherapy, immunocompromised patients due to AIDS or the like, or immunosuppressed patients due to organ transplantation or the like commonly experience a decrease in appetite, and patients after surgery may also experience anorexia.
신경성 무식욕증(또는 신경성 식욕결핍, 거식)은 척루(emaciation)에 대한 특징적 열망 및 비정상적인 식사 거동 등의 정신병성 증상뿐 아니라 표준 체중의 20% 이상의 체중 감량 및 무월경(여성의 경우) 등의 신체 증상을 보이는 질병으로, 이는 종종 젊은 여성에게 발생하며 심각하고 때로는 치명적인 질병이다. Anorexia nervosa (or anorexia nervosa, anorexia) can cause psychotic symptoms such as characteristic cravings for emaciation and abnormal eating behavior, as well as body weight loss and amenorrhea (for women) of 20% or more of standard weight. A symptomatic disease, which often occurs in young women and is a serious and sometimes fatal disease.
따라서, 식욕이 부진하거나 소화력이 약화된 경우 흔히 다양한 식욕촉진과 소화촉진을 위한 제제들이 사용되고 있다. 대표적으로는 소화효소를 다량 함유한 발효제제가 보급되고 있으며, 그 중심재료는 아밀라제, 펩티다제 등과 같은 소화효소를 다량함유한 효모를 배양하여 건조 후 타블렛화 한 것이다. 이외에, 각종 비타민과 미네랄을 다양하게 배합하여 조성한 비타민제제, 각종 소화효소제제 또는 가장 직접적으로는 식욕중추를 자극하는 제제가 사용되고 있다. 그러나, 이와 같은 약제들은 투여에 따라 직접적인 효과를 이루기는 하지만, 단순히 소화 혹은 식욕의 자극만으로 다른 장기의 활성화가 이루어지지 않은 채 식욕이 당기는 대로 섭취하게 하는 것은 오히려 질병을 악화시키는 문제점을 야기하고 있다. Therefore, when the appetite is poor or digestion is weakened, various preparations for promoting appetite and digestion are often used. Representative fermentation agents containing a large amount of digestive enzymes are widely used. The core material is a yeast containing a large amount of digestive enzymes such as amylase, peptidase, and the like, followed by drying and tableting. In addition, vitamin preparations, various digestive enzyme preparations or various formulations that stimulate the appetite center are used. However, these drugs have a direct effect upon administration, but simply ingesting the appetite without the activation of other organs by simply digesting or stimulating the appetite causes the problem of worsening the disease. .
상기와 같은 배경 하에, 본 발명자들은 효과적인 식욕 증진 또는 면역 증진 활성을 갖는 천연물을 연구하던 중, 맥아 효소액과 생약을 반응시켜 추출한 맥아 효소액 생약 추출물을 식욕억제 또는 면역억제 동물모델에 투여한 결과 장기 독성을 완화시킴과 동시에 식욕 증진 또는 면역 증진 효과를 나타내는 것을 확인함으로써 본 발명을 완성하였다.Under the above background, the present inventors, while studying natural products having an effective appetite enhancing or immune enhancing activity, the long-term toxicity as a result of administering the malt enzyme extract herbal drug extract extracted by reacting the malt enzyme solution and the herbal medicine to the appetite suppression or immunosuppression animal model The present invention was completed by confirming to alleviate the effects and at the same time exhibit an appetite enhancing or immune enhancing effect.
본 발명의 목적은 맥아 효소액을 이용하여 생체흡수율이 개선된 식욕 증진 또는 면역 증진 효과를 갖는 생약 추출물의 제조방법을 제공하는 것이다. It is an object of the present invention to provide a method for producing an herbal extract having an appetite enhancing or immune enhancing effect with improved bioabsorption using malt enzyme solution.
상기의 과제를 해결하기 위하여, 본 발명은 맥아 효소액 및 생약을 반응시키는 단계(단계 1); 및 상기 반응된 맥아 효소액 및 생약을 추출하는 단계(단계 2)를 포함하는 생약 추출물의 제조방법을 제공한다.In order to solve the above problems, the present invention comprises the steps of reacting the malt enzyme solution and the herbal medicine (step 1); And extracting the reacted malt enzyme solution and the herbal medicine (step 2).
상기 단계 1은, 상기 생약의 탄수화물 및 단백질 성분을 가수분해함으로써 저분자화하기 위하여, 맥아 효소액과 생약을 반응시키는 단계이다. 상기 맥아 효소액과 생약의 혼합비율은 50-99.9 중량%:0.1-50 중량%인 것이 바람직하다. 만약, 생약의 혼합비율이 50 중량%를 초과하는 경우, 혼합이 용이하지 않는 문제점이 발생할 수 있다. 상기 혼합은 30℃ 내지 70℃의 온도범위를 유지하면서 1시간 내지 12시간 동안 교반하여 혼합하는 것이 바람직하다. Step 1 is a step of reacting the malt enzyme solution with the herbal medicine to lower the molecular weight by hydrolyzing the carbohydrate and protein components of the herbal medicine. The mixing ratio of the malt enzyme solution and the herbal medicine is preferably 50-99.9% by weight: 0.1-50% by weight. If the mixing ratio of the herbal medicine exceeds 50% by weight, a problem may occur that mixing is not easy. The mixing is preferably mixed by stirring for 1 to 12 hours while maintaining a temperature range of 30 ℃ to 70 ℃.
본 발명에서 사용하는 용어 "맥아 효소액"은, 대맥이라 불리는 겉보리의 싹을 띄어 말린 맥아로부터 효소를 추출한 액체를 의미한다. 본 발명의 맥아 효소액은 맥아를 물에 0.1 중량% 내지 99 중량%로 첨가한 후 0.5시간 내지 24시간 혼합한 후 여과하여 제조되는 것이 바람직하다. 이때, 반응온도는 20℃ 내지 50℃가 바람직하고, 보다 바람직하게는 20℃ 내지 40℃가 바람직하다. 만약, 50℃보다 높은 온도범위에서 반응을 진행할 경우, 맥아 효소액의 효소활성이 현저히 감소하는 문제가 발생할 수 있으며, 20℃보다 낮은 온도범위에서는 맥아 효소액의 추출 수율이 떨어지는 문제가 발생할 수 있다. 이와 같은 문제점을 고려할 때, 반응온도는 실온(약 24℃)이 가장 바람직하다. The term "malt enzyme liquid" used in the present invention means a liquid from which the enzyme is extracted from the malt sprouted from the barley called dried barley. The malt enzyme solution of the present invention is preferably prepared by adding malt to water in an amount of 0.1% to 99% by weight and then mixing the mixture for 0.5 hours to 24 hours. At this time, the reaction temperature is preferably 20 ° C to 50 ° C, more preferably 20 ° C to 40 ° C. If the reaction proceeds at a temperature range higher than 50 ° C., the enzyme activity of the malt enzyme solution may be remarkably reduced, and the extraction yield of the malt enzyme liquid may drop at a temperature range lower than 20 ° C. In view of such a problem, the reaction temperature is most preferably room temperature (about 24 ° C).
상기 단계 2는, 상기 혼합된 맥아 효소액 및 생약을 추출하여 생약 추출물을 수득하기 위한 단계이다. 상기 생약은 산두근 또는 진피인 것이 바람직하다. Step 2 is a step for obtaining the herbal extract by extracting the mixed malt enzyme solution and the herbal medicine. The herbal medicine is preferably papillary or dermis.
본 발명에서 사용하는 용어 "추출물"은, 혼합된 맥아 효소액 및 생약으로부터 액체의 용매를 사용하여 추출된 특정 성분을 의미한다. 상기 용매로는 물, C1-4 알코올 또는 이들의 혼합물을 사용할 수 있다. 유효성분을 효율적으로 추출하기 위하여 에탄올로 추출하는 것이 바람직하다. 에탄올의 농도는 10% 내지 95%가 바람직하나, 이에 제한되는 것은 아니다. The term "extract" as used in the present invention means a specific component extracted from the mixed malt enzyme liquid and the crude drug using a solvent of the liquid. As the solvent, water, C 1-4 alcohol or a mixture thereof may be used. In order to efficiently extract the active ingredient, it is preferable to extract with ethanol. The concentration of ethanol is preferably 10% to 95%, but is not limited thereto.
상기 추출은 1시간 내지 24시간 동안 수행할 수 있으며, 바람직하게는 1시간 내지 5시간 동안 수행한다. 또한, 상기 추출은 환류 냉각 추출, 냉침 추출, 열수 추출 및 초음파 추출로 이루어진 군으로부터 선택된 어느 하나의 방법을 사용할 수 있으며, 유효성분을 효율적으로 추출하기 위하여 환류 냉각 추출법을 이용하는 것이 바람직하다. The extraction may be performed for 1 to 24 hours, preferably 1 to 5 hours. In addition, the extraction may use any one method selected from the group consisting of reflux cooling extraction, cold needle extraction, hot water extraction and ultrasonic extraction, it is preferable to use a reflux cooling extraction method in order to efficiently extract the active ingredient.
또한, 상기 추출물은 필요에 따라 20℃ 내지 100℃의 온도범위에서 감압농축 또는 감압 건조함으로써, 액상 또는 분말 형태로 수득할 수 있으나, 이에 제한되는 것은 아니다.In addition, the extract may be obtained in a liquid or powder form by concentrating under reduced pressure or drying under reduced pressure in the temperature range of 20 ℃ to 100 ℃ as necessary, but is not limited thereto.
본 발명에 따른 맥아 효소액을 이용한 생약 추출방법은 생약의 탄수화물 및 단백질 성분을 가수분해하여 저분자화함으로써 생체흡수율을 높이는 효과가 있다.The herbal extract method using the malt enzyme liquid according to the present invention has the effect of increasing the bioabsorption rate by hydrolyzing the carbohydrate and protein components of the herbal medicine to lower the molecular weight.
도 1은, 본 발명의 일 실시예에 따른 cisplatin으로 유도된 급성 식욕부진 동물모델에서의 맥아 효소액 산두근 추출물 및 산두근 추출물의 식욕촉진 효능 결과를 나타낸 그래프이다. 1 is a graph showing the appetite-promoting effect of malt enzyme liquid wild boar extract and wild boar extract in cisplatin-induced acute anorexia animal model according to an embodiment of the present invention.
도 2는, 본 발명의 일 실시예에 따른 cisplatin으로 유도된 급성 식욕부진 동물모델에서의 진피 추출물, 맥아 효소액 진피 추출물 및 megestrol acetate(대조약물)의 식욕촉진 효능 비교 결과를 나타낸 그래프이다. 2 is a graph showing the appetite-stimulating effect of the dermis extract, malt enzyme dermis extract and megestrol acetate (control drug) in cisplatin-induced acute anorexia animal model according to an embodiment of the present invention.
이하, 하기 제조예 및 실시예에 의하여 본 발명을 더욱 상세하게 설명하고자 한다. 단, 하기 제조예 및 실시예는 본 발명을 예시하기 위한 것일 뿐 본 발명의 범위가 이들만으로 한정되는 것은 아니다. Hereinafter, the present invention will be described in more detail by the following Preparation Examples and Examples. However, the following Preparation Examples and Examples are only for illustrating the present invention, and the scope of the present invention is not limited thereto.
실시예 1: 맥아 효소액을 이용한 산두근 추출물의 제조Example 1: Preparation of wild boar extract using malt enzyme solution
물 1 ℓ에 맥아 100 g을 넣고 1시간 동안 실온(약 24℃)에서 교반기를 이용하여 맥아 효소를 추출해낸 후, 여과하여 맥아 효소액을 수득하였다. 수득한 맥아 효소액 1 ℓ에 산두근 100 g을 분쇄하여 넣고 4시간 동안 50℃를 유지하면서 교반기를 이용하여 산두근을 맥아 효소액과 반응시켰다. 그 후, 상기 맥아 효소액 및 산두근 혼합물에 에탄올 3 ℓ를 첨가하여 75%의 에탄올을 만든 후 79℃에서 3시간 환류 냉각 추출 후 여과하여 여액을 수득하였다. 수득한 여액을 감압 농축한 후 동결건조하여 약 22 g의 산두근 추출물을 수득하였다. 100 g of malt was added to 1 L of water, and the malt enzyme was extracted using a stirrer at room temperature (about 24 ° C.) for 1 hour, followed by filtration to obtain a malt enzyme liquid. 100 g of wild bovine muscle was pulverized in 1 L of the obtained malt enzyme solution, and the wild bovine muscle was reacted with the malt enzyme liquid using a stirrer while maintaining 50 ° C. for 4 hours. Thereafter, 3 liters of ethanol was added to the malt enzyme solution and wild boar muscle mixture to make 75% ethanol, and the mixture was filtered after reflux cooling at 79 ° C. for 3 hours to obtain a filtrate. The obtained filtrate was concentrated under reduced pressure and then lyophilized to obtain about 22 g of wild boar extract.
실시예 2: 맥아 효소액을 이용한 진피 추출물의 제조Example 2: Preparation of dermis extract using malt enzyme solution
물 1 ℓ에 맥아 100 g을 넣고 1시간 동안 실온(약 24℃)에서 교반기를 이용하여 맥아 효소를 추출해낸 후, 여과하여 맥아 효소액을 수득하였다. 수득한 맥아 효소액 1 ℓ에 진피 100 g을 분쇄하여 넣고 4시간 동안 50℃를 유지하면서 교반기를 이용하여 진피를 맥아 효소액과 반응시켰다. 그 후, 상기 맥아 효소액 및 진피 혼합물에 에탄올 3 ℓ를 첨가하여 75%의 에탄올을 만든 후 79℃에서 3시간 환류 냉각 추출 후 여과하여 여액을 수득하였다. 수득한 여액을 감압 농축한 후 동결건조하여 약 30 g의 진피 추출물을 수득하였다. 100 g of malt was added to 1 L of water, and the malt enzyme was extracted using a stirrer at room temperature (about 24 ° C.) for 1 hour, followed by filtration to obtain a malt enzyme liquid. 100 g of the dermis was ground in 1 L of the obtained malt enzyme solution, and the dermis was reacted with the malt enzyme solution using a stirrer while maintaining 50 ° C for 4 hours. Thereafter, 3 l of ethanol was added to the malt enzyme solution and the dermis mixture to make 75% ethanol, and the mixture was filtered after reflux cooling at 79 ° C. for 3 hours to obtain a filtrate. The filtrate was concentrated under reduced pressure and then lyophilized to obtain about 30 g of dermis extract.
비교예 1: 산두근 추출물의 제조Comparative Example 1: Preparation of wild boar extract
산두근 100 g을 분쇄하여 75% 에탄올 1 ℓ에 침지한 후 79℃에서 3시간 동안 환류 냉각 추출한 후 여과하여 여액을 수득하였다. 수득한 여액을 감압 농축한 후 동결건조하여 약 20 g의 산두근 추출물을 수득하였다. 100 g of triceps were pulverized, immersed in 1 L of 75% ethanol, and reflux-cooled for 3 hours at 79 ° C., followed by filtration to obtain a filtrate. The obtained filtrate was concentrated under reduced pressure and then lyophilized to obtain about 20 g of wild boar extract.
비교예 2: 진피 추출물의 제조Comparative Example 2: Preparation of dermis extract
진피 100 g을 분쇄하여 75% 에탄올 1 ℓ에 침지한 후 79℃에서 3시간 동안 환류 냉각 추출한 후 여과하여 여액을 수득하였다. 수득한 여액을 감압 농축한 후 동결건조하여 약 20 g의 진피 추출물을 수득하였다. 100 g of the dermis was ground and immersed in 1 L of 75% ethanol, followed by reflux extraction at 79 ° C. for 3 hours, followed by filtration to obtain a filtrate. The filtrate was concentrated under reduced pressure and then lyophilized to obtain about 20 g of dermis extract.
실험예 1: 식욕 증진 효과 분석Experimental Example 1: Analysis of appetite enhancing effect
상기 실시예 1, 2 및 비교예 1, 2의 생약 추출물의 식욕 부진 개선 효과 확인에 앞서 비교약물인 megestrol의 식욕촉진 최고 약효 농도를 산정한 후 대조구로 사용하였다. Prior to confirming the anorexia improvement effect of the herbal extracts of Examples 1 and 2 and Comparative Examples 1 and 2, the highest appetite-stimulating concentration of megestrol as a comparative drug was calculated and used as a control.
급성 식욕저하 동물모델은 7주령의 수컷 랫트를 구입하여 1주일 간 순화기간을 거친 후 사용하였다(8주령 수컷 랫트, 250 g 내지 270 g). 각 그룹당 7마리씩 총 8그룹(정상군, cisplatin 단독 투여군, mesgentral 투여군 6)으로 분리하였고, 물을 자유로이 섭취할 수 있는 상태에서 24시간 동안 절식하였다. 세 군은 절식 18시간 후와 절식 23.5시간 후에 두 차례(bid:하루에 두 번) megestrol acetate를 0.5% methylcellulose(MC)에 희석하여 랫트에 각각 25 mg/kg, 35 mg/kg 및 45 mg/kg의 용량으로 현탁하여 경구 투여하였고, 또 다른 세 군은 절식 23.5시간 후에 한 차례(qd: 하루에 한 번) megestrol acetate를 0.5% methylcellulose(MC)에 희석하여 랫트에 각각 50 mg/kg, 70 mg/kg 및 90 mg/kg의 용량으로 현탁하여 경구 투여하였다. 절식 24시간 경과 후에 cisplatin을 랫트 몸무게 kg 당 2 mg 씩 주사용수 5 ㎖에 녹여 랫트의 복강 내 주사하였다. cisplatin 투여 직후, 한 케이지에 랫트 한 마리씩 넣고 물은 자유로이 섭취할 수 있는 상태에서 먹이를 약 50 g씩 공급해주고 기록하였다. 24시간 경과 후, 남은 먹이량을 측정하여 랫트의 먹이 섭취량을 계산하여, 각 그룹과의 먹이 섭취량 차이를 비교하였다. 그 결과를 하기 표 1에 나타내었다.Acute appetite decay animal model was used after a 7-week-old male rats after a period of 1 week of acclimatization (8-week-old male rats, 250 g to 270 g). A total of 7 rats were divided into 8 groups (normal group, cisplatin group, and mesgentral group 6) and fasted for 24 hours under free water intake. The three groups were diluted twice (bid: twice a day) megestrol acetate in 0.5% methylcellulose (MC) after 18 hours of fasting and 23.5 hours of fasting in rats 25 mg / kg, 35 mg / kg and 45 mg / Suspension was administered orally at a dose of kg, and another three groups were diluted once a day (qd: once a day) megestrol acetate in 0.5% methylcellulose (MC) 23.5 hours after fasting 50 mg / kg, 70 rats, respectively Suspension was administered orally at the dose of mg / kg and 90 mg / kg. After 24 hours of fasting, cisplatin was dissolved in 5 ml of water for injection at 2 mg / kg of rat weight and rats were injected intraperitoneally. Immediately after administration of cisplatin, one rat was placed in a cage and water was supplied freely, and 50 g of food was recorded and recorded. After 24 hours, the remaining amount of food was measured to calculate the food intake of the rats, and the difference in food intake with each group was compared. The results are shown in Table 1 below.
표 1
구분 투여량 먹이 섭취량(g/24hr)(mean SD) 먹이 섭취 증가율(%)
정상군 - 25.1±3.09 100%
cisplatin 단독 투여군 2 mg/kg cisplatin only 15.3±2.69 0%
megestrol 투여군 1 25 mg/kg(bid) megestrol acetate+2 mg/kg cisplatin 18.2±2.10 29.3%
megestrol 투여군 2 35 mg/kg(bid) megestrol acetate+2 mg/kg cisplatin 19.7±2.40 44.4%
megestrol 투여군 3 45 mg/kg(bid) megestrol acetate+2 mg/kg cisplatin 19.8±2.49 45.7%
megestrol 투여군 4 50 mg/kg(qd) megestrol acetate+2 mg/kg cisplatin 19.5±2.92 43.0%
megestrol 투여군 5 70 mg/kg(qd) megestrol acetate+2 mg/kg cisplatin 20.6±2.91 53.6%
megestrol 투여군 6 70 mg/kg(qd) megestrol acetate+2 mg/kg cisplatin 18.4±1.46 31.8%
Table 1
division Dosage Food Intake (g / 24hr) (mean SD) % Food intake growth
Normal - 25.1 ± 3.09 100%
cisplatin alone 2 mg / kg cisplatin only 15.3 ± 2.69 0%
megestrol dose group 1 25 mg / kg (bid) megestrol acetate + 2 mg / kg cisplatin 18.2 ± 2.10 29.3%
megestrol group
2 35 mg / kg (bid) megestrol acetate + 2 mg / kg cisplatin 19.7 ± 2.40 44.4%
megestrol group 3 45 mg / kg (bid) megestrol acetate + 2 mg / kg cisplatin 19.8 ± 2.49 45.7%
megestrol group 4 50 mg / kg (qd) megestrol acetate + 2 mg / kg cisplatin 19.5 ± 2.92 43.0%
megestrol group
5 70 mg / kg (qd) megestrol acetate + 2 mg / kg cisplatin 20.6 ± 2.91 53.6%
megestrol group 6 70 mg / kg (qd) megestrol acetate + 2 mg / kg cisplatin 18.4 ± 1.46 31.8%
상기 표 1에 나타난 바와 같이, 가장 효과가 좋은 하루에 한 번 megestrol acetate 70 mg/kg을 경구 투여하여 생약 추출물들과의 비교시험에 사용하였다. As shown in Table 1, oral administration of 70 mg / kg of megestrol acetate once a day for the most effective was used in comparison with the herbal extracts.
1) 산두근 추출물의 식욕 증진 효과 1) Appetite Enhancement Effect of Extract
추출방법에 따른 식욕 증진 효과를 확인하기 위하여, 상기 실시예 1에서 제조한 맥아 효소액을 이용한 산두근 추출물과 상기 비교예 1에서 제조한 산두근 추출물을 cisplatin으로 유도된 급성 식욕 저하 동물모델에 사용하여 비교실험 하였다. In order to confirm the appetite-promoting effect according to the extraction method, using the wild boiled extract using the malt enzyme solution prepared in Example 1 and the wild boiled extract prepared in Comparative Example 1 in a cisplatin-induced acute appetite depleted animal model Comparative experiment.
급성 식욕저하 동물은 7주령의 수컷 랫트를 구입하여 1주일 간 순화기간을 거친 250 g 내지 270 g의 8주령의 수컷 랫트를 사용하였다. 각 그룹당 7마리씩 4그룹(정상군, cisplatin 단독 투여군, 맥아 효소액 산두근 추출물 투여군, 산두근 추출물 투여군)으로 나누었다. 각 그룹은 물을 자유로이 섭취할 수 있는 상태에서 24시간 동안 절식시키고, 생약 추출물 투여군은 절식 19시간 후와 절식 23.5시간 후에 각각의 추출물을 0.5% methylcellulose(MC)에 희석하여 두 차례(bid: 하루에 두 번) 100 mg/kg 비교예 1의 산두근 추출물과 100 mg/kg 실시예 1의 맥아 효소액 산두근 추출물을 각각 랫트에 경구 투여하였다. 절식 24시간 경과 후에 cisplatin을 랫트 몸무게 kg 당 2 mg씩 주사용수 5 ㎖에 녹여 랫트의 복강내 주사하였다. cisplatin 투여 직후, 한 케이지에 랫트 한 마리씩 넣고 물은 자유로이 섭취할 수 있는 상태에서 먹이를 약 50 g씩 공급하고 기록하였다. 24시간 경과 후, 남은 먹이량을 측정하여 랫트의 먹이 섭취량을 계산하여, cisplatin만 단독으로 투여한 군과 각 추출물을 투여한 군의 먹이 섭취량을 비교하였다. 그 결과를 하기 표 2 및 도 1에 나타내었다. Acute anorexia animals purchased 7 week old male rats and used 250 g to 270 g 8 week old male rats that had undergone one week of acclimation. Each group was divided into 4 groups (normal group, cisplatin-only group, malt enzyme extract group extract group, group extract group). Each group fasted 24 hours under free water intake, and the herbal extract-administered group diluted each extract with 0.5% methylcellulose (MC) twice after 19 hours and 23.5 hours after fasting (bid: 1 day). Twice) 100 mg / kg wild boar extract of Comparative Example 1 and 100 mg / kg wild boil extract of malt enzyme solution of Example 1 were orally administered to rats, respectively. After 24 hours of fasting, cisplatin was dissolved in 5 ml of water for injection at 2 mg / kg of rat weight and rats were injected intraperitoneally. Immediately after administration of cisplatin, one rat was placed in a cage, and water was freely consumed. After 24 hours, the food intake of rats was calculated by measuring the amount of food remaining, and the food intakes of the cisplatin-only group and each extract-administered group were compared. The results are shown in Table 2 and FIG. 1.
표 2
구분 약물 투여량 먹이 섭취량(g/24hr)(mean SD) 먹이 섭취 증가율(%)
정상군 - (비투여) 28.3±2.6 100%
cisplatin 단독 투여군 2 mg/kg cisplatin only 7.7±2.6 Ŧ 0%
비교예 1 투여군 100 mg/kg(bid) 비교예 1 추출물+2 mg/kg cisplatin 14.6±2.2 * 33.4%
실시예 1 투여군 100 mg/kg(bid) 실시예 1 추출물+2 mg/kg cisplatin 16.3±3.1 * 41.7%
Ŧvs 정상군(p<0.05)* vs cisplatin 단독 투여군(p<0.05)
TABLE 2
division Drug dosage Food Intake (g / 24hr) (mean SD) % Food intake growth
Normal -(Non-administration) 28.3 ± 2.6 100%
cisplatin alone 2 mg / kg cisplatin only 7.7 ± 2.6 Ŧ 0%
Comparative Example 1 Administration Group 100 mg / kg (bid) Comparative Example 1 extract + 2 mg / kg cisplatin 14.6 ± 2.2 * 33.4%
Example 1 Administration Group 100 mg / kg (bid) Example 1 extract + 2 mg / kg cisplatin 16.3 ± 3.1 * 41.7%
Vs vs normal ( p <0.05) * vs cisplatin alone ( p <0.05)
상기의 표 2 및 도 1에 나타난 바와 같이, 맥아 효소액을 이용하여 추출한 실시예 1의 산두근 추출물이 식욕 증진 효능이 향상되는 것을 확인하였다. As shown in Table 2 and FIG. 1, it was confirmed that the triceps extract of Example 1 extracted using the malt enzyme solution improves appetite enhancing efficacy.
2) 진피 추출물의 식욕 증진 효과2) Appetite Enhancement Effect of Dermis Extract
추출방법에 따른 식욕 증진 효과를 확인하기 위하여, 상기 실시예 2에서 제조한 맥아 효소액을 이용한 진피 추출물과 상기 비교예 2에서 제조한 진피 추출물을 cisplatin으로 유도된 급성 식욕 저하 동물모델에 사용하여 비교실험하였다. In order to confirm the appetite enhancing effect according to the extraction method, a comparative experiment using the dermis extract using the malt enzyme solution prepared in Example 2 and the dermis extract prepared in Comparative Example 2 in a cisplatin-induced acute appetite lowered animal model It was.
급성 식욕저하 동물은 7주령의 수컷 랫트를 구입하여 1주일 간 순화기간을 거친 후 250 g 내지 270 g의 8주령의 수컷 랫트를 사용하였다. 각 그룹당 7마리씩 5그룹(정상군, cisplatin 단독 투여군, megestrol acetate 투여군, 맥아 효소액 진피 추출물 투여군, 진피 추출물 투여군)으로 나누었다. 각 그룹은 물을 자유로이 섭취할 수 있는 상태에서 24시간 동안 절식시키고, 생약 추출물 투여군은 절식 19시간 후와 절식 23.5시간 후에 각각의 추출물을 0.5% methylcellulose(MC)에 희석하여 두 차례(bid: 하루에 두 번) 600 mg/kg 비교예 2의 진피 추출물과 600 mg/kg 실시예 2의 맥아 효소액 진피 추출물을 각각 랫트에 경구 투여하였다. megestrol acetate 투여군은 앞서 측정한 것에 따라, 절식 23.5시간 후에만 한차례(qd: 하루에 한 번) 0.5 % methylcellulose(MC)에 희석하여 70 mg/kg 용량으로 경구 투여하였다. 절식 24시간 경과 후에 cisplatin을 랫트 몸무게 kg 당 2 mg씩 주사용수 5 ㎖에 녹여 랫트의 복강내 주사하였다. cisplatin 투여 직후, 한 케이지에 랫트 한 마리씩 넣고 물은 자유로이 섭취할 수 있는 상태에서 먹이를 약 50 g씩 공급하고 기록하였다. 24시간 경과 후, 남은 먹이량을 측정하여 랫트의 먹이 섭취량을 계산하여, cisplatin만 단독으로 투여한 군과 각 추출물을 투여한 군의 먹이 섭취량을 비교하였다. 그 결과를 하기 표 3 및 도 2에 나타내었다.Acute anorexia animals purchased male rats of 7 weeks of age, followed by a 1-week purifying period, and used 250- to 270 g of 8-week-old male rats. Each group was divided into 5 groups (normal group, cisplatin alone group, megestrol acetate group, malt enzyme dermal extract group, dermal extract group). Each group fasted 24 hours under free water intake, and the herbal extract-administered group diluted each extract with 0.5% methylcellulose (MC) twice after 19 hours and 23.5 hours after fasting (bid: 1 day). Twice) 600 mg / kg dermal extract of Comparative Example 2 and malt enzyme liquid dermis extract of 600 mg / kg Example 2 were orally administered to rats, respectively. As measured previously, the megestrol acetate group was orally administered at a dose of 70 mg / kg diluted in 0.5% methylcellulose (MC) once (qd: once a day) only after 23.5 hours of fasting. After 24 hours of fasting, cisplatin was dissolved in 5 ml of water for injection at 2 mg / kg of rat weight and rats were injected intraperitoneally. Immediately after administration of cisplatin, one rat was placed in a cage, and water was freely consumed. After 24 hours, the food intake of rats was calculated by measuring the amount of food remaining, and the food intakes of the cisplatin-only group and each extract-administered group were compared. The results are shown in Table 3 and FIG. 2.
표 3
구분 약물 투여량 먹이 섭취량(g/24hr)(mean SD) 먹이 섭취 증가율(%)
정상군 - (비투여) 28.3±2.5 100%
cisplatin 단독 투여군 2 mg/kg cisplatin only 13.1±2.7 Ŧ 0%
megestrol acetate 투여군 70 mg/kg(qd) megestrol acetate+2 mg/kg cisplatin 18.7±2.6 * 35.5%
비교예 2 투여군 600 mg/kg(bid) 비교예 2 추출물+2 mg/kg cisplatin 15.3±3.2 14.7%
실시예 2 투여군 600 mg/kg(bid) 실시예 2 추출물+2 mg/kg cisplatin 20.0±2.7 * 46.3%
Ŧvs 정상군(p<0.05)* vs cisplatin 단독 투여군(p<0.05)
TABLE 3
division Drug dosage Food Intake (g / 24hr) (mean SD) % Food intake growth
Normal -(Non-administration) 28.3 ± 2.5 100%
cisplatin alone 2 mg / kg cisplatin only 13.1 ± 2.7 Ŧ 0%
megestrol acetate group 70 mg / kg (qd) megestrol acetate + 2 mg / kg cisplatin 18.7 ± 2.6 * 35.5%
Comparative Example 2 Administration Group 600 mg / kg (bid) Comparative Example 2 extract + 2 mg / kg cisplatin 15.3 ± 3.2 14.7%
Example 2 Administration Group 600 mg / kg (bid) Example 2 extract + 2 mg / kg cisplatin 20.0 ± 2.7 * 46.3%
Vs vs normal ( p <0.05) * vs cisplatin alone ( p <0.05)
상기 표 3 및 도 2에 나타난 바와 같이, cisplatin 단독 투여군에서는 정상군에 비해 유의한 먹이 섭취량 감소를 보였으며, 600 mg/kg의 비교예 2의 진피 추출물을 투여한 군은 14.7%의 식욕촉진 효과를 보였다. 반면, 600 mg/kg의 실시예 2의 맥아 효소액 진피 추출물을 투여한 군은 46.3%로 31.6%의 식욕촉진 향상 효과를 확인할 수 있었다. 또한, 현재 식욕촉진제로 시판중인 대조 약물인 megestrol acetate와 동등이상의 식욕촉진 효과를 나타내는 것을 확인하였다. As shown in Table 3 and Figure 2, the cisplatin alone group showed a significant decrease in food intake compared to the normal group, 600 mg / kg group administered the dermis extract of Comparative Example 2 14.7% appetite promoting effect Showed. On the other hand, the group administered the malt enzyme liquid dermis extract of 600 mg / kg of Example 2 was confirmed that the appetite promoting effect of 31.6% to 46.3%. In addition, it was confirmed that the appetite stimulating agent exhibits an appetite-stimulating effect equal to or higher than that of megestrol acetate, a commercially available control drug.
실험예 2: 내분비계 부작용 완화 효과 분석Experimental Example 2: Analysis of endocrine side effects relief
본 발명의 맥아 효소액을 이용한 생약 추출법의 내분비계 부작용 완화 효과를 확인하기 위하여, 상기 실시예 1에서 제조한 맥아 효소액 산두근 추출물을 이용하여 cisplatin으로 유도된 만성 식욕부진 동물모델의 내분비계 부작용 완화 효과를 분석하였다. In order to confirm the endocrine system side effects mitigating effect of the herbal extract method using malt enzyme solution of the present invention, the endocrine system side effects mitigation effect of cisplatin-induced chronic anorexia animal model using malt enzyme extract triceps extract prepared in Example 1 Was analyzed.
만성 식욕부진 동물모델은 7주령의 수컷 랫트를 구입하여 1주일 간 순화기간을 거친 250 g 내지 270 g의 8주령의 수컷 랫트를 하용하였다. 각 그룹당 7마리씩 4그룹(정상군, cisplatin 단독 투여군, 실시예 1의 맥아 효소액 산두군 추출물 투여군 및 대조약물인 megestrol acetate 투여군)으로 나누었으며, 7일간 실험을 진행하였다. 실험 1일차부터 7일차까지 실시예 1의 맥아 효소액 산두근 추출물은 150 mg/kg 용량으로 오전 10시와 오후 5시 두 차례(bid: 하루에 두 번) 0.5% methylcellulose(MC)에 희석하여 랫트에 경구 투여하였고, 대조약물인 megestrol acetate는 오후 5시에만 한차례(qd: 하루에 한 번) 70 mg/kg 용량으로 0.5% methylcellulose(MC)에 희석하여 경구 투여하였다. 그 외 나머지 군들은 0.5% methylcellulose(MC)를 5 ㎖/kg의 용량으로 경구 투여하였다. 내분비계 부작용 유도를 위하여 실험 1일차와 2일차에 두 번 cisplatin을 랫트 몸무게 kg 당 2 mg씩 주사용수 5 ㎖에 녹여 랫트의 복강내 주사하였다. 실험 1일차부터, 한 케이지에 랫트 한마리씩 넣고 물은 자유로이 섭취할 수 있는 상태에서 먹이를 일정량씩 공급해주고 기록하였다. 매일 남은 먹이량과 랫트의 몸무게를 측정하고, 실험 8일차에 부검을 통해 부신(adrenal gland), 고환(testis), 신장(kidney), 간(liver), 지라(spleen)를 적출하여 cisplatin만 단독으로 투여한 그룹과 약물 투여군을 비교하였다. 결과를 하기 표 4 및 표 5에 나타내었다.The chronic anorexic animal model purchased male rats of 7 weeks old and used 250 g to 270 g of 8 week old male rats that had undergone one week of acclimation. Each group was divided into four groups (normal group, cisplatin-only group, malt-enzyme sandal group extract-administered group and control group megestrol acetate-administered group) of 7 rats, and the experiment was conducted for 7 days. From the first day to the seventh day of the experiment, the malt enzyme extract wild boiled root extract of Example 1 was diluted in 0.5% methylcellulose (MC) twice at 10 am and 5 pm (bid: twice a day) at a dose of 150 mg / kg. The control drug, megestrol acetate, was diluted orally in 0.5% methylcellulose (MC) at a dose of 70 mg / kg once (qd: once a day) only at 5 pm. The rest of the groups were orally administered 0.5 ml methylcellulose (MC) at a dose of 5 ml / kg. To induce endocrine side effects, cisplatin was dissolved twice in 5 ml of water for 2 kg / kg rats intraperitoneally and injected on the 1st and 2nd day of the experiment. From the first day of the experiment, one rat was placed in a cage, and water was supplied and recorded at a predetermined amount in a state in which water was freely ingested. Daily foods and rat weights were measured, and on day 8 of the experiment, the adrenal gland, testis, kidney, liver and spleen were removed by autopsy only cisplatin alone. Group and drug administration group were compared. The results are shown in Tables 4 and 5 below.
표 4
구분 투여량 무게(g)
체중 간 지라 신장 고환 부신
정상군 - 330.7 15.5 0.65 2.77 2.89 0.05
cisplatin 단독 투여군 2 mg/kg cisplatin only 250.6 Ŧ 11.1 Ŧ 0.60 3.27 Ŧ 2.61 Ŧ 0.04
실시예 1 투여군 150 mg/kg 실시예 1 추출물+2 mg/kg cisplatin 287.4 * 13.0 0.63 3.13 2.70 * 0.04
대조약물 투여군 70 mg/kg megestrol acetate+2 mg/kg cisplatin 275.9 13.0 0.65 3.07 Ŧ 2.70 * 0.03 *
Ŧ vs 정상군(p<0.05)* vs cisplatin 단독 투여군(p<0.05)
Table 4
division Dosage Weight (g)
weight liver Zira kidney testicle tally
Normal - 330.7 15.5 0.65 2.77 2.89 0.05
cisplatin alone 2 mg / kg cisplatin only 250.6 Ŧ 11.1 Ŧ 0.60 3.27 Ŧ 2.61 Ŧ 0.04
Example 1 Administration Group 150 mg / kg Example 1 extract + 2 mg / kg cisplatin 287.4 * 13.0 0.63 3.13 2.70 * 0.04
Reference Drug Administration Group 70 mg / kg megestrol acetate + 2 mg / kg cisplatin 275.9 13.0 0.65 3.07 Ŧ 2.70 * 0.03 *
Ŧ vs normal ( p <0.05) * vs cisplatin alone ( p <0.05)
표 5
구분 투여량 먹이 섭취량(g/24hr)(mean SD) 먹이 섭취 증가율(%)
정상군 - 23.3±3.0 100%
cisplatin 단독 투여군 2 mg/kg cisplatin only 7.4±4.6 Ŧ 0%
실시예 1 투여군 150 mg/kg 실시예 1 추출물+2 mg/kg cisplatin 14.8±3.6 * 46.8%
대조약물 투여군 70 mg/kg megestrol acetate+2 mg/kg cisplatin 14.8±5.6 * 46.6%
Ŧ vs 정상군(p<0.05)* vs cisplatin 단독 투여군(p<0.05)
Table 5
division Dosage Food Intake (g / 24hr) (mean SD) % Food intake growth
Normal - 23.3 ± 3.0 100%
cisplatin alone 2 mg / kg cisplatin only 7.4 ± 4.6 Ŧ 0%
Example 1 Administration Group 150 mg / kg Example 1 extract + 2 mg / kg cisplatin 14.8 ± 3.6 * 46.8%
Reference Drug Administration Group 70 mg / kg megestrol acetate + 2 mg / kg cisplatin 14.8 ± 5.6 * 46.6%
Ŧ vs normal ( p <0.05) * vs cisplatin alone ( p <0.05)
상기 표 4 및 표 5에 나타난 바와 같이, cisplatin 2회 단독 투여한 cisplatin 단독 투여군에서는 정상군 대비 몸무게, 먹이 섭취량에서 유의한 감소를 보였으며, 장기무게에서는 간, 신장, 고환에서 유의한 변화를 보이는 것을 확인하였다. 이는 cisplatin 투여가 단순한 식욕부진으로 인한 먹이 섭취량 감소와 그로 인한 몸무게 저하를 초래하는 것 뿐 만아니라, 내분비계 관련 장기들에도 직ㆍ간접적으로 부작용을 초래하는 결과로 볼 수 있다. 반면, 실시예 1의 맥아 효소액 산두근 추출물 150 mg/kg을 투여한 군은 cisplatin 단독 투여군보다 먹이 섭취량이 유의하게 증진되었으며, 이에 따른 몸무게의 저하도 완화될 수 있음을 확인하였다. 또한, cisplatin 투여에 의한 간의 무게 감소 부작용도 완화되었고, 간과 고환도 마찬가지로 cisplatin 투여에 의한 부작용을 완화해주는 것을 확인할 수 있었다. 상기의 결과에 나타난 바와 같이, 맥아 효소액을 이용하여 추출한 산두근 추출물은 식욕촉진 효능 뿐만 아니라, 내분비계 관련 장기들의 부작용도 완화시킬 수 있음을 확인하였다. 또한, 현재 시판중인 대조약물인 megestrol acetate와 비교하였을 경우에도 동등이상의 식욕 촉진 효과는 물론 내분비계 장기들의 부작용 완화효과도 있음을 확인하였다. As shown in Table 4 and Table 5, cisplatin alone-administered cisplatin alone showed a significant decrease in weight and food intake compared to normal, and showed significant changes in liver, kidney and testes in long-term weight. It was confirmed. Not only does cisplatin administration lead to a decrease in food intake and weight loss due to anorexia, but also direct and indirect side effects on endocrine organs. On the other hand, the group administered 150 mg / kg of malt enzyme liquid wild boar extract of Example 1 was significantly improved food intake than the cisplatin alone group, it was confirmed that the decrease in weight can be alleviated. In addition, the side effects of cisplatin-induced weight loss were alleviated, and the liver and testicles were similarly alleviated. As shown in the above results, it was confirmed that the extract of wild boar muscle using malt enzyme solution can alleviate side effects of endocrine-related organs as well as appetite-stimulating effect. In addition, when compared with the current commercial control drug megestrol acetate, it was confirmed that the appetite-stimulating effect is higher than the equivalent, as well as the side effects of endocrine organs.
실험예 3: 면역억제 완화 효과 분석Experimental Example 3: Analysis of Immunosuppressive Mitigation Effect
상기 실시예 1에서 제조한 맥아 효소액을 이용한 산두근 추출물의 면역억제 완화 효과를 확인하기 위하여, cyclophosphamide로 유도된 면역억제 동물모델 사용하여 분석하였다. In order to confirm the immunosuppressive mitigation effect of the extract of wild boar muscle using the malt enzyme solution prepared in Example 1, it was analyzed using a cyclophosphamide-induced immunosuppressive animal model.
면역억제 동물모델은 7주령의 수컷 랫트를 구입하여 1주일 간의 순화기간을 거친 250 g 내지 270 g의 8주령의 수컷 랫트를 사용하였다. 랫트는 각 7마리씩 4그룹(정상군, cyclophosphanide 단독 투여군, 실시예 1의 맥아 효소액 산두근 추출물 투여군 및 대조약물 megestrol acetate 투여군)으로 분리하고 6일간 실험을 진행하였다. 실험 1일차부터 6일차까지 실시예 1의 맥아 효소액 산두근 추출물 투여군은 오전 10시와 오후 5시에 실시에 1의 맥아 효소액 산두근 추출물을 150 mg/kg씩 두 차례(bid: 하루에 두 번) 0.5% methylcellulose(MC)에 희석하여 경구 투여하였으며, 대조약물 megestrol 투여군은 하루에 한 번 오후 5시에 0.5% methylcellulose(MC)에 희석하여 70 mg/kg megestrol acetate 용량으로 경구 투여 하였다. 그 외 나머지 군은 0.5% methylcellulose(MC)를 5 ㎖/kg으로 경구 투여하였다. 면역억제 유도를 위해 실험 4일차에 한 번 cyclophosphamide를 랫트 몸무게 kg 당 25 mg씩 주사용수 5 ㎖에 녹인 후 복강내 주사하였다. 실험 1일차부터, 한 케이지에 랫트 한마리씩 넣고 물과 먹이는 자유로이 섭취할 수 있도록 한 상태에서 매일 몸무게를 측정하였으며, 실험 7일차에 부검을 통해 지라(spleen)와 흉선(thymus)을 적출하여 cyclophosphamide 단독으로 투여한 군과 비교하였다. 결과를 하기 표 6에 나타내었다.The immunosuppressive animal model used male rats of 7 weeks old and used 8-week old male rats ranging from 250 g to 270 g after one week of acclimation. The rats were separated into four groups (normal group, cyclophosphanide alone group, malt enzyme solution of wild bovine muscle extract group of Example 1 and control group megestrol acetate group) of 7 rats, and the experiment was conducted for 6 days. From day 1 to day 6 of the experiment, the maltose extract wild boiled root extract group of Example 1 was carried out at 10 am and 5 pm at 150 mg / kg twice the malt enzyme boiled bovine root extract of 1 (bid: twice a day). ) Dilution in 0.5% methylcellulose (MC) was administered orally, and the control drug megestrol group was diluted in 0.5% methylcellulose (MC) at 5 pm once a day orally administered at a dose of 70 mg / kg megestrol acetate. The rest of the groups were orally administered with 0.5 ml methylcellulose (MC) at 5 ml / kg. To induce immunosuppression, cyclophosphamide was once dissolved in 5 ml of water for injection at 25 mg / kg of rats once every 4 days of the experiment and injected intraperitoneally. From the first day of the experiment, each rat was placed in a cage so that the water and the food were freely ingested, and the weight was measured every day. On the seventh day, the spleen and the thymus were removed by autopsy, and cyclophosphamide alone was used. Compared with the group administered. The results are shown in Table 6 below.
표 6
구분 투여량 무게(g) 체중대비 %
체중 지라 흉선
정상군 - 279.7 0.58±0.04 0.55±0.04
cyclophosphamide 단독 투여군 25 mg/kg cyclophosphamide 277.0 0.40±0.04 Ŧ 0.25±0.04 Ŧ
실시예 1 투여군 150 mg/kg 실시예1 추출물+25 mg/kg cyclophosphamide 281.6 0.47±0.14 0.30±0.06
대조약물 투여군 70 mg/kg megestrol acatate+25 mg/kg cyclophosphamide 276.3 0.42±0.06 0.20±0.05
Ŧ vs 정상군(p>0.05)
Table 6
division Dosage Weight (g) % Of body weight
weight Zira Thymus
Normal - 279.7 0.58 ± 0.04 0.55 ± 0.04
cyclophosphamide alone 25 mg / kg cyclophosphamide 277.0 0.40 ± 0.04 Ŧ 0.25 ± 0.04 Ŧ
Example 1 Administration Group 150 mg / kg Example 1 extract + 25 mg / kg cyclophosphamide 281.6 0.47 ± 0.14 0.30 ± 0.06
Reference Drug Administration Group 70 mg / kg megestrol acatate + 25 mg / kg cyclophosphamide 276.3 0.42 ± 0.06 0.20 ± 0.05
Ŧ vs normal group ( p > 0.05)
상기 표 6에 나타난 바와 같이, cyclophosphamide 단독 투여군의 경우 정상군과 비교하여 몸무게의 변화는 없었으나, 면역세포 관련 장기인 지라와 흉선 무게에서는 유의한 감소를 나타내었다. 이는 cyclophosphamide 투여로 인해 면역이 억제되었음을 나타내는 것이다. 반면, 실시예 1의 추출물 투여군에서는 cyclophosphamide 단독 투여군과 비교하여 몸무게 변화는 나타내지 않았으나, 지라와 흉선 모두에서 cyclophosphamide로 유도된 면역장기의 무게감소를 완화하는 효능을 나타내었다.As shown in Table 6, the cyclophosphamide alone group showed no change in body weight compared to the normal group, but showed a significant decrease in splenic and thymus weight, which is an immune cell-related organ. This indicates that immunity was suppressed by cyclophosphamide administration. On the other hand, the extract administration group of Example 1 showed no change in body weight compared to the cyclophosphamide alone group, but showed an effect of alleviating weight loss of cyclophosphamide-induced immune organs in both spleen and thymus.

Claims (7)

  1. 맥아 효소액 및 생약을 반응시키는 단계; 및Reacting the malt enzyme solution and the herbal medicine; And
    상기 혼합된 맥아 효소액 및 생약을 추출하는 단계를 포함하는Extracting the mixed malt enzyme solution and herbal medicine
    생약 추출물의 제조방법.Method of preparing herbal extracts.
  2. 제1항에 있어서, 상기 맥아 효소액 및 생약은 1시간 내지 12시간 동안 혼합하여 반응시키는 것을 특징으로 하는 제조방법.The method according to claim 1, wherein the malt enzyme solution and the herbal medicine are reacted by mixing for 1 hour to 12 hours.
  3. 제1항에 있어서, 상기 맥아 효소액은 물 및 맥아를 0.5시간 내지 24시간 혼합한 후 여과하여 제조되는 것을 특징으로 하는 제조방법.The method according to claim 1, wherein the malt enzyme solution is prepared by mixing water and malt for 0.5 to 24 hours and then filtering.
  4. 제1항에 있어서, 상기 맥아 효소액 및 생약 혼합은 30℃ 내지 70℃ 온도범위에서 수행하는 것을 특징을 하는 제조방법.The method according to claim 1, wherein the malt enzyme and the herbal mixture are mixed at 30 ° C to 70 ° C.
  5. 제1항에 있어서, 상기 맥아 효소액 및 생약의 혼합비율은 50-99.9 중량%:0.1-50 중량%인 것을 특징으로 하는 제조방법.The method of claim 1, wherein the mixing ratio of the malt enzyme liquid and the herbal medicine is 50-99.9% by weight: 0.1-50% by weight.
  6. 제1항에 있어서, 상기 추출용매는 물, C1-4 알코올 또는 이들의 혼합용매인 것을 특징으로 하는 제조방법.The method of claim 1, wherein the extraction solvent is water, C 1-4 alcohol or a mixed solvent thereof.
  7. 제1항에 있어서, 상기 생약은 산두근 또는 진피인 것을 특징으로 하는 제조방법.The method of claim 1, wherein the herbal medicine is a papillary muscle or dermis.
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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20020003165A (en) * 2001-11-23 2002-01-10 송윤강 Process for Producing a fermented food for health aid
KR20030053417A (en) * 2001-12-22 2003-06-28 함종천 Health food composition which contains Saururus chinensis Baill. and others
KR20100130265A (en) * 2009-06-03 2010-12-13 정세헌 A manufacturing method of functional foods for natural drugstuff and thereby natural drugstuff
KR20130045659A (en) * 2011-10-26 2013-05-06 (주)함소아 Oriental medicine composition for chidren's anorexia and manufacturing method thereof

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20020003165A (en) * 2001-11-23 2002-01-10 송윤강 Process for Producing a fermented food for health aid
KR20030053417A (en) * 2001-12-22 2003-06-28 함종천 Health food composition which contains Saururus chinensis Baill. and others
KR20100130265A (en) * 2009-06-03 2010-12-13 정세헌 A manufacturing method of functional foods for natural drugstuff and thereby natural drugstuff
KR20130045659A (en) * 2011-10-26 2013-05-06 (주)함소아 Oriental medicine composition for chidren's anorexia and manufacturing method thereof

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