WO2013077634A1 - Composition for preventing and treating non-alcoholic fatty liver disease comprising cordyceps militaris extract - Google Patents

Composition for preventing and treating non-alcoholic fatty liver disease comprising cordyceps militaris extract Download PDF

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WO2013077634A1
WO2013077634A1 PCT/KR2012/009890 KR2012009890W WO2013077634A1 WO 2013077634 A1 WO2013077634 A1 WO 2013077634A1 KR 2012009890 W KR2012009890 W KR 2012009890W WO 2013077634 A1 WO2013077634 A1 WO 2013077634A1
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liver disease
fatty liver
alcoholic fatty
extract
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박형진
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Park Hyoung Jin
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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
    • A61K36/06—Fungi, e.g. yeasts
    • A61K36/062—Ascomycota
    • A61K36/066—Clavicipitaceae
    • 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
    • A61K36/06—Fungi, e.g. yeasts
    • A61K36/062—Ascomycota
    • A61K36/066—Clavicipitaceae
    • A61K36/068—Cordyceps
    • 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
    • A61K36/06—Fungi, e.g. yeasts
    • A61K36/07—Basidiomycota, e.g. Cryptococcus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics

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  • the present invention provides a composition effective for the prevention and treatment of non-alcoholic steatosis comprising an extract of Cordyceps militaris.
  • the present invention also provides a method for extracting the extract.
  • Non-alcoholic fatty liver disease is a condition in which triglycerides accumulate in the liver regardless of drinking, and include nonalcoholic fatty liver disease (steatosis) and nonalcoholic steatohepatitis (steatohepatitis).
  • Non-alcoholic fatty liver disease is classified into macrovesicular fatty liver disease and microvesicular fatty liver disease according to the pathological size of fat droplets present in liver cells. Most non-alcoholic steatohepatic is macrovesicular. In the early stages of the onset, several small fat droplets appear in the liver cells, but as the fat droplets become larger and larger, the large vesicles are made. ring) shape. Nonalcoholic fatty liver disease is chronically progressing slowly, so there are generally no specific symptoms, but liver function gradually worsens, such as the activity of alkaline phosphatase or aminotransferase in blood, which is a measure of liver function.
  • fatty acids migrate from adipose tissue to liver due to deficiencies in fat metabolism due to an imbalance in energy consumption.
  • a decrease in receptor molecules (PPAR- ⁇ PPAR- ⁇ and SREBP1) that regulate the action of enzymes required for the oxidation or synthesis of fatty acids is one cause of fat accumulation in the liver.
  • Fatty liver disease which has severe fat accumulation, progresses to inflammation, steatohepatitis, which increases NF ⁇ B activation and tumor necrotizing factor- ⁇ expression due to oxidative stress.
  • Hepatic necrosis occurs in livers with extensive fatty hepatitis, and stellate cells are activated, resulting in fibrosis, especially perisinusoidal fibrosis at the end of hepatic veins. Therefore, hepatitis is improved by removing the cause of fat accumulation, but develops into fibrosis or cirrhosis of the liver if left unchecked. Liver fibrosis or cirrhosis occurs in 15-50% of patients with nonalcoholic steatohepatitis, and cirrhosis develops after 10 years in 30% of fibrotic patients.
  • Nonalcoholic fatty liver disease is highly associated with obesity or diabetes. Since both obese and diabetic patients have been increasing rapidly in recent years, nonalcoholic fatty liver disease has emerged as a hot spot in modern society. The incidence of nonalcoholic steatohepatitis increases significantly from 10 to 15% in people with normal weight and to 80% in people with overweight. The incidence of nonalcoholic steatohepatitis increases significantly from 10 to 15% in people with normal weight and to 80% in people with overweight. In 2005, the prevalence of obesity in Korean adults was 31.5% over 20 years old and 34.8% over 30 years old. In particular, the prevalence of obesity over the age of 20 increased from 26.3% in 1998 to 31.5% in 2005, and over 30 years increased from 29.1% to 34.8%.
  • Nonalcoholic fatty liver disease is usually recovered by proper intake of nutrients and control of related metabolic diseases. However, if left untreated, hepatitis is followed by fibrosis or hardening of the liver, resulting in poor prognosis. Therefore, in order to prevent the progression of non-alcoholic fatty liver disease to cirrhosis, it is necessary to prevent the accumulation of fat in the liver.However, to prevent the accumulation of fat in the liver, there are other treatment options besides reducing calorie intake or increasing calorie consumption through exercise. This is not presented.
  • Cordyceps militaris is a type of mushroom that grows naturally based on the pupa of moths. Extract of pupa cordyceps sinensis inhibits liver fibrosis (Nan et al, Arch Pharm Res 24: 327-332, 2001) and alleviates liver damage caused by liver toxic substances (Kim et al., Journal of Oriental Physiology and Pathology 16: 684 -692, 202), or repair of hepatocyte damage by ethanol (Yan et al, Afr J Biotech 7: 4004-4009, 2008) has been reported. However, the effect of pupa cordyceps sinensis on non-alcoholic fatty liver disease in which liver fat accumulation and resulting liver function decreases is not known yet.
  • the present invention provides a composition for the prevention and treatment of non-alcoholic fatty liver disease, including the extract of pupa cordyceps sinensis.
  • the extract is selected from the group consisting of:
  • the composition inhibits the accumulation of fat in the liver in experimental animals reared for 12 weeks in a high fat diet (containing 60% fat).
  • the composition inhibits an increase in the activity of alanine aminotransferase or asparate aminotransferase in the blood in experimental animals reared for 12 weeks in a high fat diet.
  • the present invention also provides an agent for the prevention or treatment of non-alcoholic fatty liver disease comprising the composition and a pharmaceutically acceptable carrier.
  • the present invention also provides an agent for the prevention or treatment of alcoholic fatty liver disease comprising the composition and a pharmaceutically acceptable carrier.
  • the present invention provides a dietary supplement or functional food for the prevention or improvement of non-alcoholic fatty liver disease comprising the composition and the carrier.
  • the present invention also provides an agent for the prevention or treatment of alcoholic fatty liver disease comprising the composition and a pharmaceutically acceptable carrier.
  • compositions of the present invention have been shown to inhibit fat accumulation in the liver caused by high fat diet and improve liver function. Therefore, the composition of the present invention can be usefully used for the prevention and treatment of nonalcoholic fatty liver disease.
  • FIG. 1 is a photograph staining the liver tissue of the experimental animal by hematoxylin-eosin method.
  • NC Normal control
  • FC high fat diet control group
  • Cm high fat diet and pupa cordyceps extract administration group.
  • Figure 2 shows the liver function test results of the experimental animal.
  • FC experimental animals
  • ALT, A alanine aminotransferase
  • AST, B asparate aminotransferase
  • NC normal animals
  • the present invention provides a composition that effectively prevents the accumulation of fat in hepatocytes, and thus acts effectively in the prevention and treatment of nonalcoholic fatty liver disease.
  • the present invention also provides pharmaceutical compositions and foods comprising the composition.
  • composition of the present invention is characterized in that it comprises an extract extracted from pupa cordyceps.
  • the extract of chrysalis cordyceps sinensis is a step of extracting hot water from chrysalis cordyceps; ii) mixing the hot water extract with cold ethanol to obtain a precipitate and iii) dissolving the precipitate in distilled water followed by freeze drying.
  • fruiting body powder of pupa cordyceps is used. This is for obtaining the mushroom extract more efficiently because when the fruiting body powder is used to increase the extraction surface area, more active ingredients can be extracted during the extraction process.
  • the fruiting body powder is put in distilled water and boiled. At this time, by boiling over 3 hours at a temperature of 100 °C ⁇ 104 °C to ensure that the active ingredient is sufficiently extracted.
  • the extract thus obtained is filtered to remove solid components and then concentrated under reduced pressure to increase the content of the active ingredients.
  • the concentrated concentrate under reduced pressure is mixed with cold ethanol (-20 ⁇ ⁇ ).
  • the extract-ethanol mixture is left overnight at a temperature below 4 ° C. and then centrifuged to obtain a precipitate. Finally, the precipitate is again dissolved in distilled water sufficiently and then lyophilized.
  • the alanine aminotransferase and asparate aminotransferase activity in the blood is increased in the experimental animals bred only 12 weeks high fat diet.
  • alanine aminotransferase and asparate aminotransferase in blood were significantly decreased in experimental animals fed with oral chrysalis extract daily during high-fat diet. Therefore, the extract can be confirmed to improve the liver function associated with fatty liver disease.
  • the description of the present invention describes an embodiment for fatty liver disease caused by a high fat diet.
  • the composition comprising the extract of pupa cordyceps sinensis of the present invention is not limited to the prevention and treatment of non-alcoholic fatty liver disease, it can also be used for the prevention and treatment of alcoholic fatty liver disease.
  • the composition is particularly useful for metabolic diseases such as obesity, diabetes mellitus, Cushing's syndrome; Ulcerative colitis, chronic pancreatitis, ileal bypass surgery, prolonged parenteral nutrition as diseases or treatments causing malnutrition; Or it can be used for the prevention and treatment of non-alcoholic fatty liver disease caused by various liver toxins and the like.
  • compositions may be administered orally or parenterally.
  • Oral formulations form the compositions in pill, capsule or liquid form
  • parenteral formulations form the composition in liquid form.
  • the formulations together with the compositions include various excipients, carriers, diluents and the like known to those of ordinary skill in the art.
  • the fruiting body powder of pupa cordyceps was added to distilled water, boiled for 3 hours, filtered to remove solids, and the extract was concentrated under reduced pressure.
  • the concentrated extract was mixed with cold ethanol (-20 ° C.) and left overnight at 4 ° C. or lower.
  • the ethanol mixture was centrifuged to give a precipitate which was dissolved in distilled water again and then lyophilized to obtain a powder.
  • the animals were fed on a high-fat diet for 12 weeks, anesthetized, collected from the heart, and the livers were extracted and fixed in 4% paraformaldehyde solution. Immobilized liver tissue was embedded with paraffin, and 5 ⁇ m-thick serial sections were prepared. Liver sections were stained with hematoxylin-eosin (H & E) method and observed for fat accumulation of hepatic inclusions. The fat accumulated in the macrovesicles of hepatocytes was eliminated during staining by H & E method, so the macrovesicles were empty and rounded.
  • H & E hematoxylin-eosin
  • Figure 1 presents the effect of pupa cordyceps sinensis extract on high fat diet-induced fatty liver disease in experimental animals. Giant vesicles were not observed in livers of control animals (NCs) fed normal diets, whereas large vesicles were observed in livers of experimental animals (FCs) raised for 12 weeks using only high-fat diets. In addition, it was confirmed that the number of macrovesicles in the liver cells was significantly reduced in the experimental animals (Cm), which were orally administered with the chrysalis cordyceps extract daily, while providing a high fat diet compared to the experimental animals (FC).
  • NCs control animals
  • FCs experimental animals
  • alanine aminotransferase and asparate aminotransferase which are indicators of liver function, in serum isolated from blood of experimental animals was measured using an automatic analyzer.
  • ALT, A alanine aminotransferase
  • AST, B asparate aminotransferase
  • FC experimental animals
  • NC normal control animals
  • FC blood alanine aminotransferase and asparate aminotransferase activities were significantly lower in experimental animals (Cm), which were fed orally administered pupae Cordyceps sinensis extract daily, while being fed in high fat diet compared to experimental animals (FC).

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Abstract

The present invention relates to a composition comprising an extract of Cordyceps militaris. In test animals, the composition comprising the extract of Cordyceps militaris improved liver function impairment or reduced fat accumulation in liver cells induced by long-term feeding on a high-fat diet. Consequently, the composition of the present invention can be used to advantage in preventing or treating alcoholic fatty liver disease and non-alcoholic fatty liver disease brought on by various causes.

Description

번데기동충하초 추출물을 포함하는 비알코올성 지방간증의 예방 및 치료용 조성물Composition for the prevention and treatment of non-alcoholic fatty liver disease comprising pupa cordyceps sinensis extract
본 발명은 번데기동충하초(Cordyceps militaris)의 추출물을 포함하는 비알코올성 지방간증(non-alcoholic steatosis)의 예방 및 치료에 유효한 조성물을 제공한다. 또한 본 발명은 상기 추출물을 추출하는 방법을 제공한다.The present invention provides a composition effective for the prevention and treatment of non-alcoholic steatosis comprising an extract of Cordyceps militaris. The present invention also provides a method for extracting the extract.
지방간질환(fatty liver disease)은 과다한 알코올 섭취로 인한 알코올성 지방간질환과 그렇지 않은 비알코올성 지방간질환으로 구분된다. 비알코올성 지방간질환은 음주와 관계없이 간 내에 중성지방이 축적되는 질환으로 여기에는 비알코올성 지방간증(steatosis)과 비알코올성 지방간염(steatohepatitis)이 포함된다. Fatty liver disease is classified into alcoholic fatty liver disease due to excessive alcohol intake and nonalcoholic fatty liver disease. Non-alcoholic fatty liver disease is a condition in which triglycerides accumulate in the liver regardless of drinking, and include nonalcoholic fatty liver disease (steatosis) and nonalcoholic steatohepatitis (steatohepatitis).
비알코올성 지방간증은 병리학적으로 간세포에 출현하는 지방 방울의 크기에 따라 거대소포성 지방간증과 미세소포성 지방간증으로 구분된다. 대부분의 비알코올성 지방간증은 거대소포성으로 발병 초기에는 간세포 내에 여러 개의 작은 지방 방울이 출현하나 진행하면서 지방 방울이 점점 커지면서 거대소포가 만들어 지며 거대소포가 핵을 세포 중앙부에서 한쪽으로 밀어내어 도장반지(signet ring) 모양을 나타낸다. 비알코올성 지방간증은 만성적으로 서서히 진행되므로 일반적으로 특이한 증상이 없으나 간 기능의 척도인 혈중 alkaline phosphatase 또는 aminotransferase의 활성도가 상승하는 등 간 기능이 점차 나빠진다.Non-alcoholic fatty liver disease is classified into macrovesicular fatty liver disease and microvesicular fatty liver disease according to the pathological size of fat droplets present in liver cells. Most non-alcoholic steatohepatic is macrovesicular. In the early stages of the onset, several small fat droplets appear in the liver cells, but as the fat droplets become larger and larger, the large vesicles are made. ring) shape. Nonalcoholic fatty liver disease is chronically progressing slowly, so there are generally no specific symptoms, but liver function gradually worsens, such as the activity of alkaline phosphatase or aminotransferase in blood, which is a measure of liver function.
비알코올성 지방간증에서는 에너지 소모의 불균형에 의한 지방대사의 결함으로 인해 지방조직으로부터 간으로 이동하는 지방산이 증가한다. 또한 지방산의 산화 또는 합성에 필요한 효소의 작용을 조절하는 수용체 분자(PPAR-α PPAR-γ 및 SREBP1)의 감소가 간 내 지방축적의 한 원인이다. 지방축적이 심한 지방간증은 염증 즉 지방간염(steatohepatitis)으로 진행되는데 이때에는 oxidative stress로 인한 NFκB의 활성화와 tumor necrotizing factor-α의 발현이 증가한다. In nonalcoholic fatty liver disease, fatty acids migrate from adipose tissue to liver due to deficiencies in fat metabolism due to an imbalance in energy consumption. In addition, a decrease in receptor molecules (PPAR-α PPAR-γ and SREBP1) that regulate the action of enzymes required for the oxidation or synthesis of fatty acids is one cause of fat accumulation in the liver. Fatty liver disease, which has severe fat accumulation, progresses to inflammation, steatohepatitis, which increases NFκB activation and tumor necrotizing factor-α expression due to oxidative stress.
광범위한 지방간염을 수반하는 간에서는 간세포의 괴사가 나타나며 또한 성상세포(stellate cell)가 활성화되어 섬유화가 일어나게 되는데 특히 간정맥 말단 부위에서 굴모양혈관주변섬유화(perisinusoidal fibrosis)가 심하게 나타난다. 따라서 지방간염은 지방이 축적하는 원인을 제거하면 개선되나 방치하면 간의 섬유화 또는 경변으로 발전한다. 비알코올성 지방간염 환자의 15 - 50%에서 간의 섬유화 또는 경변이 나타나며 섬유화 환자의 30%에서는 10년 후에 간경변이 나타난다. Hepatic necrosis occurs in livers with extensive fatty hepatitis, and stellate cells are activated, resulting in fibrosis, especially perisinusoidal fibrosis at the end of hepatic veins. Therefore, hepatitis is improved by removing the cause of fat accumulation, but develops into fibrosis or cirrhosis of the liver if left unchecked. Liver fibrosis or cirrhosis occurs in 15-50% of patients with nonalcoholic steatohepatitis, and cirrhosis develops after 10 years in 30% of fibrotic patients.
비알코올성 지방간증은 비만증 또는 당뇨병과 관련이 매우 높다. 비만증 환자와 당뇨병 환자 모두가 최근 들어 급격히 증가하는 추세이므로 비알코올성 지방간증은 현대 사회에서 주목 받는 질환으로 대두되고 있다. 비알코올성 지방간증의 발병률은 정상체중을 지닌 사람에서는 10 - 15%이나 과체중을 지닌 사람에서는 80%로 크게 상승한다. 비알코올성 지방간증의 발병률은 정상체중을 지닌 사람에서는 10 - 15%이나 과체중을 지닌 사람에서는 80%로 크게 상승한다. 2005년에 국내 성인 비만의 유병률은 20세 이상에서 31.5%, 30세 이상에서 34.8%이었다. 특히 20세 이상의 비만 유병률은 1998년의 26.3%에서 2005년에 31.5%로 증가하였으며, 30세 이상은 29.1%에서 34.8%로 증가하였다. Nonalcoholic fatty liver disease is highly associated with obesity or diabetes. Since both obese and diabetic patients have been increasing rapidly in recent years, nonalcoholic fatty liver disease has emerged as a hot spot in modern society. The incidence of nonalcoholic steatohepatitis increases significantly from 10 to 15% in people with normal weight and to 80% in people with overweight. The incidence of nonalcoholic steatohepatitis increases significantly from 10 to 15% in people with normal weight and to 80% in people with overweight. In 2005, the prevalence of obesity in Korean adults was 31.5% over 20 years old and 34.8% over 30 years old. In particular, the prevalence of obesity over the age of 20 increased from 26.3% in 1998 to 31.5% in 2005, and over 30 years increased from 29.1% to 34.8%.
비알코올성 지방간증은 영양물질의 적절한 섭취와 유관 대사성 질환의 조절에 의해 대개 회복된다. 그러나 방치하면 지방간염에 이어 간의 섬유화 또는 경화로 진행되어 예후가 불량해 진다. 따라서 비알코올성 지방간증이 간경변으로 진행하는 것을 차단하기 위해서는 간에 지방이 축적하는 것을 예방해야 하나 아직까지 간의 지방 축적을 예방하기 위해서는 칼로리 섭취를 줄이거나 운동으로 칼로리 소모를 증가시키는 방법 외에는 이렇다 할 치료방안이 제시되어 있지 않다. Nonalcoholic fatty liver disease is usually recovered by proper intake of nutrients and control of related metabolic diseases. However, if left untreated, hepatitis is followed by fibrosis or hardening of the liver, resulting in poor prognosis. Therefore, in order to prevent the progression of non-alcoholic fatty liver disease to cirrhosis, it is necessary to prevent the accumulation of fat in the liver.However, to prevent the accumulation of fat in the liver, there are other treatment options besides reducing calorie intake or increasing calorie consumption through exercise. This is not presented.
한편 번데기동충하초(Cordyceps militaris)는 나방류의 번데기를 기주로 하여 자생하는 버섯의 일종이다. 번데기동충하초의 추출물이 간 섬유화를 억제하며(Nan et al, Arch Pharm Res 24:327-332, 2001), 간 독성물질로 유발한 간 손상을 경감시키며(김 등, 동의생리병리학회지 16권: 684-692, 202), 또는 에탄올에 의한 간세포의 손상을 회복시킨다(Yan et al, Afr J Biotech 7:4004-4009, 2008)는 연구결과가 보고되어 있다. 그러나 번데기동충하초가 간 내의 지방축적 및 그로 인한 간 기능이 저하하는 비알코올성 지방간증에 미치는 효과에 대해서는 아직까지 알려진 바가 없다.Cordyceps militaris, on the other hand, is a type of mushroom that grows naturally based on the pupa of moths. Extract of pupa cordyceps sinensis inhibits liver fibrosis (Nan et al, Arch Pharm Res 24: 327-332, 2001) and alleviates liver damage caused by liver toxic substances (Kim et al., Journal of Oriental Physiology and Pathology 16: 684 -692, 202), or repair of hepatocyte damage by ethanol (Yan et al, Afr J Biotech 7: 4004-4009, 2008) has been reported. However, the effect of pupa cordyceps sinensis on non-alcoholic fatty liver disease in which liver fat accumulation and resulting liver function decreases is not known yet.
본 발명은 번데기동충하초 추출물을 포함하는 비알코올성 지방간증의 예방 및 치료에 유효한 신규한 조성물을 제공하는 것을 목적으로 한다. It is an object of the present invention to provide a novel composition effective for the prevention and treatment of non-alcoholic steatohepatitis comprising pupa cordyceps sinensis extract.
본 발명은 번데기동충하초의 추출물을 포함하는 비알코올성 지방간증의 예방 및 치료용 조성물을 제공한다.The present invention provides a composition for the prevention and treatment of non-alcoholic fatty liver disease, including the extract of pupa cordyceps sinensis.
바람직하게, 상기 추출물은 Preferably, the extract
i) 번데기동충하초를 열수 추출하는 단계;i) hot water extracting pupa cordyceps;
ii) 상기 열수 추출물을 냉 에탄올과 혼합하여 침전물을 취하는 단계 및ii) mixing the hot water extract with cold ethanol to obtain a precipitate, and
iii) 상기 침전물을 증류수에 용해시킨 후 동결 건조하는 단계로부터 얻어지는 것이다.iii) obtained by dissolving the precipitate in distilled water and freeze drying.
바람직하게, 상기 조성물은 고지방식이(지방을 60% 함유)로 12주 동안 사육한 실험동물에서 간 내의 지방축적을 억제한다.Preferably, the composition inhibits the accumulation of fat in the liver in experimental animals reared for 12 weeks in a high fat diet (containing 60% fat).
바람직하게, 상기 조성물은 고지방식이로 12주 동안 사육한 실험동물에서 혈중 alanine aminotransferase 또는 asparate aminotransferase의 활성의 상승을 억제한다.Preferably, the composition inhibits an increase in the activity of alanine aminotransferase or asparate aminotransferase in the blood in experimental animals reared for 12 weeks in a high fat diet.
또한 본 발명은 상기 조성물 및 약제학적으로 허용되는 담체를 포함하는 비알코올성 지방간증의 예방 또는 치료용 제재를 제공한다.The present invention also provides an agent for the prevention or treatment of non-alcoholic fatty liver disease comprising the composition and a pharmaceutically acceptable carrier.
또한 본 발명은 상기 조성물 및 약제학적으로 허용되는 담체를 포함하는 알코올성 지방간증의 예방 또는 치료용 제재를 제공한다.The present invention also provides an agent for the prevention or treatment of alcoholic fatty liver disease comprising the composition and a pharmaceutically acceptable carrier.
또한 본 발명은 상기 조성물 및 담체를 포함하는 비알코올성 지방간증의 예방 또는 개선용 건강보조 식품 또는 기능성 식품을 제공한다.In another aspect, the present invention provides a dietary supplement or functional food for the prevention or improvement of non-alcoholic fatty liver disease comprising the composition and the carrier.
또한 본 발명은 상기 조성물 및 약제학적으로 허용되는 담체를 포함하는 알코올성 지방간증의 예방 또는 치료용 제재를 제공한다.The present invention also provides an agent for the prevention or treatment of alcoholic fatty liver disease comprising the composition and a pharmaceutically acceptable carrier.
본 발명의 조성물은 고지방식이로 유발한 간 내의 지방 축적을 억제하고 간 기능을 개선하는 것으로 나타났다. 따라서 본 발명의 조성물은 비알코올성 지방간증의 예방 및 치료에 유용하게 활용될 수 있다. The compositions of the present invention have been shown to inhibit fat accumulation in the liver caused by high fat diet and improve liver function. Therefore, the composition of the present invention can be usefully used for the prevention and treatment of nonalcoholic fatty liver disease.
도1은 실험동물의 간 조직을 hematoxylin-eosin 법으로 염색한 사진이다. 고지방식이만을 12주 동안 제공한 실험동물의 간(FC)에서는 간 세포 내의 지방축적을 의미하는 원형의 거대소포가 다수 출현하며, 이러한 고지방식이에 의한 거대소포의 출현이 번데기동충하초 추출물의 투여(Cm)에 의해 현저히 감소한다. NC; 정상대조군, FC: 고지방식이 대조군, Cm: 고지방식이 및 번데기동충하초 추출물 투여군. 1 is a photograph staining the liver tissue of the experimental animal by hematoxylin-eosin method. In the livers of experimental animals provided with only high fat diet for 12 weeks, a large number of circular vesicles indicating fat accumulation in liver cells appeared, and the appearance of giant vesicles by high fat diet was administered by pupa extract Markedly reduced by (Cm). NC; Normal control, FC: high fat diet control group, Cm: high fat diet and pupa cordyceps extract administration group.
도2는 실험동물의 간 기능 검사결과를 도시한 것이다. 고지방식이만을 12주 동안 제공한 실험동물(FC)의 혈액에서는 alanine aminotransferase(ALT, A) 및 asparate aminotransferase(AST, B)의 활성이 정상동물(NC)에 비해 크게 상승하며, 이러한 고지방식이에 의한 간 기능 검사치의 상승이 번데기동충하초 추출물의 투여(Cm)에 의해 현저히 개선된다. 별표(*): 통계적으로 유의함을 표시, NC; 정상대조군, FC: 고지방식이 대조군, Cm: 고지방식이 및 번데기동충하초 추출물 투여군. Figure 2 shows the liver function test results of the experimental animal. In the blood of experimental animals (FC), which had been fed only high-fat diet for 12 weeks, the activity of alanine aminotransferase (ALT, A) and asparate aminotransferase (AST, B) increased significantly compared to normal animals (NC). The elevation of the liver function test by is significantly improved by administration of pupa cordyceps sinensis extract (Cm). Asterisk (*): indicate statistically significant, NC; Normal control, FC: high fat diet control group, Cm: high fat diet and pupa cordyceps extract administration group.
본 발명은 간세포 내의 지방축적을 방지하여 비알코올성 지방간증의 예방 및 치료에 유효하게 작용하는 조성물을 제공한다. 본 발명은 또한 상기 조성물을 포함하는 약제학적 조성물 및 식품을 제공한다.The present invention provides a composition that effectively prevents the accumulation of fat in hepatocytes, and thus acts effectively in the prevention and treatment of nonalcoholic fatty liver disease. The present invention also provides pharmaceutical compositions and foods comprising the composition.
본 발명의 조성물은 번데기동충하초로부터 추출된 추출물을 포함하는 것을 특징으로 한다. The composition of the present invention is characterized in that it comprises an extract extracted from pupa cordyceps.
본 발명에서 상기 번데기동충하초의 추출물은 i) 번데기동충하초를 열수 추출하는 단계; ii) 상기 열수 추출물을 냉 에탄올과 혼합하여 침전물을 취하는 단계 및 iii) 상기 침전물을 증류수에 용해시킨 후 동결 건조하는 단계로부터 얻어진다.In the present invention, the extract of chrysalis cordyceps sinensis is a step of extracting hot water from chrysalis cordyceps; ii) mixing the hot water extract with cold ethanol to obtain a precipitate and iii) dissolving the precipitate in distilled water followed by freeze drying.
본 발명에서는 번데기동충하초의 자실체 가루를 이용한다. 이는 버섯 추출물을 보다 효율적으로 얻기 위한 것으로 자실체 가루를 이용할 경우 추출 표면적이 넓어지므로, 추출 과정에서 보다 많은 유효 성분들이 추출될 수 있기 때문이다. In the present invention, fruiting body powder of pupa cordyceps is used. This is for obtaining the mushroom extract more efficiently because when the fruiting body powder is used to increase the extraction surface area, more active ingredients can be extracted during the extraction process.
다음으로, 자실체 가루의 추출물을 얻기 위해 자실체 가루를 증류수에 넣고 끓이는 과정을 거친다. 이때 100℃ ~ 104℃의 온도에서 3시간 이상에 걸쳐 끓임으로써 유효 성분이 충분히 추출되도록 한다. Next, in order to obtain an extract of the fruiting body powder, the fruiting body powder is put in distilled water and boiled. At this time, by boiling over 3 hours at a temperature of 100 ℃ ~ 104 ℃ to ensure that the active ingredient is sufficiently extracted.
이렇게 얻은 추출액을 여과하여 고형성분을 제거한 다음 감압 농축하여 유효 성분의 함량을 높인다. 감압 농축된 추출액을 냉 에탄올(-20℃)과 혼합한다. 상기 추출액-에탄올 혼합물을 4℃ 이하의 온도에서 하룻밤 동안 방치한 다음 원심분리하여 침전물을 취한다. 마지막으로, 상기 침전물을 다시 증류수에 충분히 용해한 다음 동결건조 한다. The extract thus obtained is filtered to remove solid components and then concentrated under reduced pressure to increase the content of the active ingredients. The concentrated concentrate under reduced pressure is mixed with cold ethanol (-20 占 폚). The extract-ethanol mixture is left overnight at a temperature below 4 ° C. and then centrifuged to obtain a precipitate. Finally, the precipitate is again dissolved in distilled water sufficiently and then lyophilized.
실험동물을 고지방식이로 장기간 사육하면 지방간증이 유발되는데 이렇게 유발된 지방간증은 인간의 비알코올성 지방간증과 유사한 병리학적 및 면역학적 소견을 나타내므로(Park et al, J Hepatol (2011),doi:10.1016/j.jhep2010.08.019), 본 발명에서는 이 실험동물 모델을 이용하여 번데기동충하초의 추출물이 비알코올성 지방간증에 유효함을 입증한다. Long-term rearing of experimental animals in a high-fat diet leads to fatty liver disease, which has similar pathological and immunological findings as human nonalcoholic fatty liver disease (Park et al, J Hepatol (2011), doi : 10.1016 / j.jhep2010.08.019), The present invention demonstrates that the extract of pupa cordyceps sinensis is effective for nonalcoholic fatty liver disease using this experimental animal model.
본 발명에서 실험동물을 12주 동안 고지방식이만(지방을 60% 함유)으로 사육한 실험동물에서는 간세포 내의 지방축적이 크게 증가한다. 그러나 고지방식이로 사육하는 동안 번데기동충하초 추출물을 매일 경구 투여한 실험동물에서는 고지방식이만으로 사육한 실험동물에 비해 간세포 내의 지방축적이 현저히 감소한다. 따라서 상기 추출물은 실험동물에서 고지방식이로 유발한 지방간증의 발생을 억제하는 것을 확인할 수 있다.In the present invention, in the experimental animals in which the experimental animals were raised only by high fat diet (containing 60% fat) for 12 weeks, fat accumulation in hepatocytes is greatly increased. However, the fat accumulation in hepatocytes was significantly reduced in the experimental animals fed orally administered pupa cordyceps extract daily during the high fat diet, compared to the experimental animals raised only by the high fat diet. Therefore, the extract can be confirmed to suppress the occurrence of fatty liver disease caused by a high fat diet in the experimental animal.
또한 본 발명에서 실험동물을 12주 동안 고지방식이만으로 사육한 실험동물에서는 혈중 alanine aminotransferase 그리고 asparate aminotransferase의 활성이 상승한다. 그러나 고지방식이로 사육하는 동안 번데기동충하초 추출물을 매일 경구 투여한 실험동물에서는 고지방식이만으로 사육한 실험동물에 비해 혈중 alanine aminotransferase 그리고 asparate aminotransferase가 현저히 감소한다. 따라서 상기 추출물은 지방간증에 수반되는 간의 기능저하를 향상시키는 것을 확인할 수 있다. In addition, in the present invention, the alanine aminotransferase and asparate aminotransferase activity in the blood is increased in the experimental animals bred only 12 weeks high fat diet. However, alanine aminotransferase and asparate aminotransferase in blood were significantly decreased in experimental animals fed with oral chrysalis extract daily during high-fat diet. Therefore, the extract can be confirmed to improve the liver function associated with fatty liver disease.
본 발명에 대한 설명에서는 고지방식이로 유발한 지방간증에 대한 실시예를 기재한다. 그러나 본 발명의 번데기동충하초의 추출물을 포함하는 조성물은 비알코올성 지방간증의 예방 및 치료에만 한정되는 것이 아니라 알코올성 지방간증의 예방 및 치료에도 사용될 수 있다. The description of the present invention describes an embodiment for fatty liver disease caused by a high fat diet. However, the composition comprising the extract of pupa cordyceps sinensis of the present invention is not limited to the prevention and treatment of non-alcoholic fatty liver disease, it can also be used for the prevention and treatment of alcoholic fatty liver disease.
상기 조성물은 특히 비만, 당뇨병, Cushing씨 증후군 등의 대사성 질환; 영양결핍을 일으키는 질환 또는 치료로서 궤양성 결장염, 만성 췌장염, 공회장우회술, 장기간의 전비경구적영양; 또는 각종 간독소 등으로 유발되는 비알코올성 지방간증의 예방 및 치료에 사용될 수 있다. The composition is particularly useful for metabolic diseases such as obesity, diabetes mellitus, Cushing's syndrome; Ulcerative colitis, chronic pancreatitis, ileal bypass surgery, prolonged parenteral nutrition as diseases or treatments causing malnutrition; Or it can be used for the prevention and treatment of non-alcoholic fatty liver disease caused by various liver toxins and the like.
이러한 조성물은 경구 또는 비경구적으로 투여될 수 있다. 경구용은 조성물을 알약, 캡슐 또는 액상의 형태로 제형화 하며, 비경구용은 조성물을 액상으로 제형화 한다. 제형에는 조성물과 함께 본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자에게 알려진 다양한 부형제, 담체, 희석제 등이 포함된다.Such compositions may be administered orally or parenterally. Oral formulations form the compositions in pill, capsule or liquid form, and parenteral formulations form the composition in liquid form. The formulations together with the compositions include various excipients, carriers, diluents and the like known to those of ordinary skill in the art.
이하 실시예를 통해 본 발명을 상세히 설명한다. 그러나 이는 발명의 이해를 돕기 위한 것일 뿐, 본 발명이 이에 한정되는 것으로 여겨져서는 안 된다. The present invention will be described in detail through the following examples. However, this is only to aid the understanding of the invention, the present invention should not be considered to be limited thereto.
실시예EXAMPLE
[번데기동충하초로부터 추출물의 제조]  [Preparation of Extract from Chrysalis Cordyceps sinensis]
번데기동충하초의 자실체 가루를 증류수에 넣고 3시간 끓인 다음 여과하여 고형성분을 제거하고 추출액을 감압 농축하였다. 상기 농축 추출액을 냉 에탄올(-20℃)과 혼합하고 4℃ 이하에서 하룻밤 동안 방치했다. 에탄올 혼합물을 원심분리하여 침전물을 취하고 그것을 다시 증류수에 용해한 다음 동결건조하여 분말을 얻었다. The fruiting body powder of pupa cordyceps was added to distilled water, boiled for 3 hours, filtered to remove solids, and the extract was concentrated under reduced pressure. The concentrated extract was mixed with cold ethanol (-20 ° C.) and left overnight at 4 ° C. or lower. The ethanol mixture was centrifuged to give a precipitate which was dissolved in distilled water again and then lyophilized to obtain a powder.
[실험동물에서 지방간증의 유발 및 번데기동충하초 추출물 투여][Induction of Fatty Liver Disease and Administration of Chrysalis Cordyceps Sinensis Extract] in Experimental Animals
6-7주령의 C57BL/6 마우스 수컷을 실온 및 습도가 조절되는 환경에 1주일 동안 적응시켰다. 지방간증을 유발하기 위해 실험동물을 고지방식이(조지방 60% 함유)로 12주 동안 사육했다. 반면 정상대조동물은 일반사료(조지방 4.5% 함유)로 사육했다. 번데기동충하초 추출물을 증류수에 용해한 다음 고지방식이로 사육하기 시작한 첫날부터 실험이 끝날 때까지 매일 1회씩 체중당 100 mg/Kg씩 경구투여 했다. 6-7 week old C57BL / 6 mouse males were acclimated for one week in a controlled environment of room temperature and humidity. To induce fatty liver disease, experimental animals were raised for 12 weeks in a high fat diet (containing 60% crude fat). Normal control animals, on the other hand, were reared with regular feed (containing 4.5% crude fat). Pupa Cordyceps sinensis extract was dissolved in distilled water and then administered orally at a daily dose of 100 mg / Kg once a day until the end of the experiment.
[간 내의 지방축적에 대한 효과 관찰][Observation of effect on fat accumulation in liver]
번데기동충하초 추출물이 고지방식이로 유발한 간 내의 지방축적에 미치는 효과를 다음과 같이 관찰했다. The effects of pupa cordyceps sinensis extract on fat accumulation in liver induced by high fat diet were observed as follows.
고지방식이로 12주간 사육한 실험동물을 마취하고 심장에서 채혈한 다음 간을 적출하여 4% paraformaldehyde 용액에 고정했다. 고정된 간 조직을 paraffin으로 포매하고 5 ㎛ 두께의 연속절편을 제작했다. 간 절편을 hematoxylin-eosin(H & E) 법으로 염색한 다음 간세 내포의 지방축적 상태를 관찰했다. 간세포의 거대소포에 축적된 지방은 H & E 법으로 염색하는 과정에서 탈락되므로 거대소포는 비어있고 둥근 형태를 나타내었다. The animals were fed on a high-fat diet for 12 weeks, anesthetized, collected from the heart, and the livers were extracted and fixed in 4% paraformaldehyde solution. Immobilized liver tissue was embedded with paraffin, and 5 μm-thick serial sections were prepared. Liver sections were stained with hematoxylin-eosin (H & E) method and observed for fat accumulation of hepatic inclusions. The fat accumulated in the macrovesicles of hepatocytes was eliminated during staining by H & E method, so the macrovesicles were empty and rounded.
도1에 실험동물에서 고지방식이로 유발된 지방간증에 대한 번데기동충하초 추출물의 효과를 제시하였다. 고지방식이만으로 12주 동안 사육한 실험동물(FC)의 간에서 거대소포가 다수 관찰되었음에 비해 정상식이로 사육한 대조동물(NC)의 간에서는 거대소포가 관찰되지 않다. 또한 고지방식이를 제공하면서 번데기동충하초 추출물을 매일 경구 투여한 실험동물(Cm)에서는 고지방식이만으로 사육한 실험동물(FC)에 비해 간 세포 내의 거대소포의 수가 크게 감소되었음이 확인되었다. Figure 1 presents the effect of pupa cordyceps sinensis extract on high fat diet-induced fatty liver disease in experimental animals. Giant vesicles were not observed in livers of control animals (NCs) fed normal diets, whereas large vesicles were observed in livers of experimental animals (FCs) raised for 12 weeks using only high-fat diets. In addition, it was confirmed that the number of macrovesicles in the liver cells was significantly reduced in the experimental animals (Cm), which were orally administered with the chrysalis cordyceps extract daily, while providing a high fat diet compared to the experimental animals (FC).
이러한 결과는 번데기동충하초 추출물이 고지방식이로 유발된 지방간증에서 간 내의 지방축적을 현저히 억제함을 나타낸다.These results indicate that pupa cordyceps sinensis extract significantly inhibits fat accumulation in the liver in high fat diet-induced fatty liver disease.
[간 기능에 대한 효과 관찰] [Observing effect on liver function]
번데기동충하초 추출물이 고지방식이로 유발한 지방간증에서 간 기능 저하에 미치는 효과를 다음과 같이 관찰했다. The effects of pupa cordyceps sinensis extract on liver function deterioration in high fat diet-induced fatty liver disease were observed as follows.
실험동물의 혈액으로부터 분리한 혈청에서 간 기능의 지표인 alanine aminotransferase와 asparate aminotransferase의 활성을 자동분석기를 사용해 측정했다. The activity of alanine aminotransferase and asparate aminotransferase, which are indicators of liver function, in serum isolated from blood of experimental animals was measured using an automatic analyzer.
도2에 실험동물에서 고지방식이로 유발한 지방간증의 간 기능에 대한 번데기동충하초 추출물의 효과를 제시하였다. 고지방식이로 12주 동안 사육한 실험동물(FC)의 혈액에서는 alanine aminotransferase(ALT, A) 및 asparate aminotransferase(AST, B)의 활성이 정상식이로 사육한 정상대조동물(NC)에 비해 크게 상승했다. 또한 고지방식이로 사육하면서 번데기동충하초 추출물을 매일 경구 투여한 실험동물(Cm)에서는 고지방식이만으로 사육한 실험동물(FC)에 비해 혈중 alanine aminotransferase 와 asparate aminotransferase 활성이 현저히 낮아졌음이 확인되었다. 2 shows the effect of pupa cordyceps sinensis extract on liver function of fatty liver disease induced by high fat diet in experimental animals. The activity of alanine aminotransferase (ALT, A) and asparate aminotransferase (AST, B) was significantly higher in the blood of experimental animals (FC) raised for 12 weeks on a high-fat diet compared to normal control animals (NC). did. In addition, it was confirmed that blood alanine aminotransferase and asparate aminotransferase activities were significantly lower in experimental animals (Cm), which were fed orally administered pupae Cordyceps sinensis extract daily, while being fed in high fat diet compared to experimental animals (FC).
이상의 결과는 번데기동충하초 추출물이 고지방식이로 유발된 지방간증에서 간 기능 저하를 현저히 개선함을 나타낸다.The above results indicate that pupa cordyceps sinensis extract significantly improves liver function in fatty liver disease induced by high fat diet.

Claims (12)

  1. 번데기동충하초(Cordyceps militaris)의 추출물을 포함하는 것을 특징으로 하는 비알코올성 지방간증의 예방 및 치료용 조성물.Chrysalis Cordyceps sinensis (Cordyceps militaris) composition for the prevention and treatment of non-alcoholic fatty liver disease, characterized in that it comprises an extract.
  2. 제1항에서,In claim 1,
    상기 추출물은 The extract is
    i) 번데기동충하초를 열수 추출하는 단계;i) hot water extracting pupa cordyceps;
    ii) 상기 열수 추출물을 냉 에탄올과 혼합하여 침전물을 취하는 단계 및ii) mixing the hot water extract with cold ethanol to obtain a precipitate, and
    iii) 상기 침전물을 증류수에 용해시킨 후 동결 건조하는 단계로부터 얻어지는 것을 특징으로 하는 비알코올성 지방간증의 예방 및 치료용 조성물.iii) a composition for the prevention and treatment of non-alcoholic fatty liver disease, which is obtained by dissolving the precipitate in distilled water and freeze drying.
  3. 제1항에서,In claim 1,
    상기 조성물은 간 내의 지방축적을 억제하는 것을 특징으로 하는 비알코올성 지방간증의 예방 및 치료용 조성물.The composition is a composition for preventing and treating non-alcoholic fatty liver disease, characterized in that it inhibits the accumulation of fat in the liver.
  4. 제1항에서,In claim 1,
    상기 조성물은 비알코올성 지방간증으로 유발된 혈중 alanine aminotransferase 및 asparate aminotransferase 활성의 상승을 억제하는 것을 특징으로 하는 비알코올성 지방간증의 예방 및 치료용 조성물.The composition is a composition for the prevention and treatment of non-alcoholic fatty liver disease, characterized in that to suppress the rise of alanine aminotransferase and asparate aminotransferase activity in the blood induced by non-alcoholic fatty liver disease.
  5. 제1항 내지 제4항 중 어느 한 항의 조성물 및 약제학적으로 허용되는 담체를 포함하는 비알코올성 지방간증 및 비알코올성 지방간염의 예방 및 치료용 제재.An agent for the prevention and treatment of nonalcoholic steatohepatitis and nonalcoholic steatohepatitis comprising the composition of any one of claims 1 to 4 and a pharmaceutically acceptable carrier.
  6. 제5항에서,In claim 5,
    상기 제재는 비만, 당뇨병 또는 Cushing씨 증후군의 대사성 질환; 영양결핍을 일으키는 질환 또는 치료로서 궤양성 결장염, 만성 췌장염, 공회장우회술 또는 장기간의 전비경구적영양; 또는 간독소로 인해 유발되는 비알코올성 지방간증 및 비알코올성 지방간염의 예방 및 치료용 제재.The agent is a metabolic disease of obesity, diabetes or Cushing's syndrome; Ulcerative colitis, chronic pancreatitis, ileal bypass surgery or prolonged parenteral nutrition as diseases or treatments causing malnutrition; Or agents for preventing and treating nonalcoholic steatohepatitis and nonalcoholic steatohepatitis caused by hepatoxin.
  7. 제5항에서,In claim 5,
    상기 제재는 정제, 캡슐 또는 경구용 또는 비경구용 액상인 것을 특징으로 하는 비알코올성 지방간증 및 비알코올성 지방간염의 예방 및 치료용 제재.The agent is a tablet, capsule or oral or parenteral liquid for the prevention and treatment of non-alcoholic steatohepatitis and non-alcoholic steatohepatitis, characterized in that.
  8. 제1항 내지 제4항 중 어느 한 항의 조성물 및 약제학적으로 허용되는 담체를 포함하는 알코올성 지방간증 및 알코올성 지방간염의 예방 및 치료용 제재.An agent for the prevention and treatment of alcoholic fatty liver disease and alcoholic fatty hepatitis, comprising the composition of any one of claims 1 to 4 and a pharmaceutically acceptable carrier.
  9. 제8항에서,In claim 8,
    상기 제재는 정제, 캡슐 또는 경구용 또는 비경구용 액상인 것을 특징으로 하는 알코올성 지방간증 및 알코올성 지방간염의 예방 및 치료용 제재.The agent is an agent for the prevention and treatment of alcoholic fatty liver disease and alcoholic fatty hepatitis, characterized in that the tablet, capsule or oral or parenteral liquid.
  10. 제1항 내지 제4항 중 어느 한 항의 조성물 및 담체를 포함하는 비알코올성 지방간증 및 비알코올성 지방간염의 예방 및 개선용 건강보조 또는 기능성 식품.A health supplement or functional food for preventing and improving nonalcoholic steatohepatitis and nonalcoholic steatohepatitis comprising the composition and carrier of any one of claims 1 to 4.
  11. 제10항에서,In claim 10,
    상기 식품은 비만, 당뇨병 또는 Cushing씨 증후군의 대사성 질환; 영양결핍을 일으키는 질환 또는 치료로서 궤양성 결장염, 만성 췌장염, 공회장우회술 또는 장기간의 전비경구적영양; 또는 간독소로 인해 유발되는 비알코올성 지방간증 및 비알코올성 지방간염의 예방 및 개선용 건강보조 또는 기능성 식품. The food products are metabolic diseases of obesity, diabetes or Cushing's syndrome; Ulcerative colitis, chronic pancreatitis, ileal bypass surgery or prolonged parenteral nutrition as diseases or treatments causing malnutrition; Or a health supplement or functional food for preventing and improving nonalcoholic fatty liver disease and nonalcoholic steatohepatitis caused by hepatoxin.
  12. 제1항 내지 제4항 중 어느 한 항의 조성물 및 담체를 포함하는 알코올성 지방간증 및 알코올성 지방간염의 예방 및 개선용 건강보조 또는 기능성 식품.A health supplement or functional food for preventing and improving alcoholic fatty liver disease and alcoholic fatty hepatitis, comprising the composition and carrier of any one of claims 1 to 4.
PCT/KR2012/009890 2011-11-25 2012-11-21 Composition for preventing and treating non-alcoholic fatty liver disease comprising cordyceps militaris extract WO2013077634A1 (en)

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