KR20210030192A - Pharmaceutical composition containing complex Rubus crataegifolius, Crataegus pinnatifida Bunge, Cinnamomum cassia extracts for prevention or treatment of metabolic syndrome - Google Patents

Abstract

A complex extract of a Rubus crataegifolius extract, a Crataegus pinnatifida extract and/or a Cinnamomum cassia extract of the present invention can significantly reduce the amount of food and dietary hormone and leptin and significantly increase adiponectin to reduce body weight and body fat weight, can reduce liver weight and the level of liver enzymes AST and ALT, can reduce hepatic lipid and the size and number of fat globules in fatty liver tissue, can significantly reduce the activity of hepatic lipid metabolism-related enzymes, and can also have effects of significantly increasing beta-oxidation-related CPT activity, lowering blood sugar, overcoming glucose tolerance, inhibiting blood clotting and platelet aggregation, and significantly inhibiting arteriosclerosis, in an obese animal model compared to a single extract. Thus the complex extract can be advantageously used for the prevention, treatment or amelioration of metabolic syndrome diseases.

Description

Pharmaceutical composition containing complex Rubus crataegifolius, Crataegus pinnatifida Bunge, Cinnamomum cassia extracts for prevention or treatment of metabolic syndrome }

The present invention relates to a pharmaceutical composition for preventing or treating metabolic syndrome disease.

Metabolic syndrome refers to a syndrome in which risk factors such as hypertriglyceridemia, hypertension, glucose metabolism, fatty liver, obesity, and blood coagulation disorders appear together. This metabolic syndrome or metabolic syndrome is a conception of a phenomenon in which risk factors for various cardiovascular diseases and type 2 diabetes form clusters into one disease group. It is a useful concept that can comprehensively explain insulin resistance and related complex and various metabolic abnormalities and clinical manifestations. Having metabolic syndrome increases the risk of developing cardiovascular disease or type 2 diabetes.

This symptom has been known for a long time, but in 1988 Reaven argued that the common cause of these symptoms is insulin resistance, which does not work well in the body, and named it X(x) syndrome and insulin resistance syndrome. In 1998, the World Health Organization decided to call it “metabolic syndrome” instead of the term “insulin resistance syndrome” because there was no evidence that insulin resistance could explain all of these symptoms. The diagnostic criteria suggested by the National Cholesterol Education Program (NCEP) of the United States are as follows.

Abdominal obesity: Waist circumference of male 102 cm (Asian 90 cm), female 88 cm (Asian 80 cm) or more

Triglyceride: 150 mg/dr or more

High-density cholesterol: male 40md/dr, female 50 mg/dr

Fasting blood sugar: 110 mg/dr or more or during diabetes treatment

Blood pressure: systolic 130 mmHg or higher or diastolic 85 mmHg or higher

If three or more of these five indicators exceed the threshold, it is considered metabolic syndrome.

Insulin resistance was first pointed out as the cause of metabolic syndrome. To lower blood pressure, excess insulin secreted cannot regulate blood pressure, induces vascular cell proliferation, thickens the wall of blood vessels, promotes lipolysis, increases triglycerides, and increases HDL- It is known that metabolic syndrome, such as hypertension, hyperlipidemia, and abdominal obesity, occurs overall by lowering cholesterol.

With the recent increase in adult diseases, vascular disorders such as arteriosclerosis are increasing significantly. Among them, thrombosis occurs from a mechanism in which platelets, one of the blood components, are deposited when a thrombus formed by coagulation of blood in the cardiovascular and blood vessels in a living body undergoes physiological damage while flowing through the vascular system. In vivo, blood coagulation and dissolution are always in balance, and blood clots do not form during normal circulation. However, when the balance is disturbed due to various pathological factors, the blood flow becomes impaired and cardiovascular disease occurs due to this. Therefore, the occurrence of functional abnormalities and circulatory abnormalities in the blood causes various vascular diseases, and when these functions and circulation disorders appear in the brain and heart, it is fatal to life, and the sequelae thereof is also serious. When looking at the statistical results of death and cause of death in Korea, cancer, cerebrovascular disease and heart disease were in order (Statistics Office, 2007 Statistical Yearbook).

Because metabolic syndrome increases the risk of cardiovascular disease and diabetes, once diagnosed, active treatment should be initiated to reduce the risk of developing these diseases. The primary treatment is to have a healthy lifestyle, limit calories, increase exercise and change diet so that you can achieve 5-10% weight loss per year. If the improvement of lifestyle is not enough, or if the risk of cardiovascular disease is very high, you can use an insulin sensitizer to treat medication. However, prevention is very important because a clear way to treat it has not yet been identified. Obesity is the most fundamental cause, so it should be prevented with proper weight maintenance and regular exercise. In addition, it is important to relax your mind by controlling your mental and physical environment to avoid stress.

Republic of Korea Patent Publication No. 10-2016-0132611

It is an object of the present invention to provide a pharmaceutical composition for preventing or treating metabolic syndrome diseases.

Another object of the present invention is to provide a health functional food composition for preventing or improving metabolic syndrome diseases.

In order to achieve the above object, the present invention is raspberry ( Rubus crataegifolius ) extract, hawthorn ( Crataegus) It provides a pharmaceutical composition for preventing or treating metabolic syndrome diseases containing a mixture of pinnatifida) extract and cinnamon ( Cinnamomum cassia) extract as an active ingredient.

In addition, the present invention raspberry ( Rubus crataegifolius ) extract, hawthorn ( Crataegus pinnatifida ) extract and cinnamon ( Cinnamomum cassia ) provides a health functional food composition for preventing or improving metabolic syndrome disease containing a mixture of the extract as an active ingredient.

Raspberries of the present invention ( Rubus crataegifolius ) extract, hawthorn ( Crataegus pinnatifida ) extract and/or cinnamon ( Cinnamomum cassia ) extract complex extract significantly reduced dietary amount, dietary hormone and leptin in an obese animal model than a single extract, and adiponectin Significantly increases, can reduce body weight and body fat weight, can reduce liver weight and liver enzymes AST and ALT levels, can reduce liver lipids in fatty liver tissue and reduce the size and number of fat cells, liver lipids Significantly decrease the activity of metabolism-related enzymes, and also significantly increase the activity of CPT involved in beta-oxidation, lower blood sugar in blood, overcome glucose tolerance, inhibit blood coagulation and inhibit platelet aggregation, and arteries Since it exhibits an effect of remarkably inhibiting hardening, it can be usefully used for preventing, treating or improving metabolic syndrome diseases.

1 is a normal diet (ND), a high fat diet (HFD), a high fat diet + Simvastin (S, SMV), a high fat diet in a metabolic syndrome prevention experiment using highly obese inducing animals (C57BL/6N mice). Lee + Raspberry Unripe Fruit (HFD+R), High Fat Diet + Hawthorn (HFD+C), High Fat Diet + Raspberry + Hawthorn (HFD+R+C), High Fat Diet + Raspberry + Hawthorn + Cinnamon (HFD+R) +C+C') is a graph showing the results of the change in body weight during the experiment (1a) and the body weight (1b) and fat weight (1c) after the experiment.
Figure 1d is a high-fat diet (HFD) + raspberries + hawthorn (HFD + R + C) or high-fat diet + raspberries + hawthorn + cinnamon (HFD + R + C + C') when treated with a decrease in the amount of diet. It is a graph showing a decrease in leptin and an increase in adiponectin due to.
Figure 2 is when treated with ND, HFD, HFD+R, HFD+C, HFD+R+C, HFD+R+C+C' in the metabolic syndrome prevention experiment using highly obese inducing animals (C57BL/6N mice) It is a graph showing the results of changes in liver weight (2a) and liver ALT, AST (2b, 2c) activity.
Figure 3 is when treated with ND, HFD, HFD+R, HFD+C, HFD+R+C, HFD+R+C+C' in a metabolic syndrome prevention experiment using highly obese inducing animals (C57BL/6N mice) Shows the experimental results on the change of total cholesterol (TC) (3a), triglyceride (TG) (3b), HDL-cholesterol (HDL-C) (3c) and LDL-cholesterol (LDL-C) (3d) of It is a graph.
Figure 4 is a normal diet (ND), high fat diet (HFD) and HFD+R, HFD+C, HFD+R+C, and HFD+R in a metabolic syndrome prevention experiment using highly obese inducing animals (C57BL/6N mice). It is a graph showing the activity of adipogenesis-regulating enzymes (FAS, HMG-CoAR/lipid synthesis and CPT/lipolysis) when treated with +C+C'.
Figure 5 shows ND, HFD, HFD+Simvastin, HFD+R, HFD+R+C, HFD+R+C+C' in a metabolic syndrome prevention experiment using highly obese inducing animals (C57BL/6N mice). This is a photograph showing the results of histological analysis (H&E-staining; 5a, Oil Red-staining; 5b) of liver tissue after treatment.
6 is a normal diet (ND), high fat diet (HFD) and HFD+R, HFD+C, HFD+R+C, and HFD+R+ in metabolic syndrome experiments using highly obese inducing animals (C57BL/6 mice). After administration of a high fat diet (HFD) for 8 weeks in C+C', the level of fasting and postprandial glucose was measured after inducing diabetes and maintaining a diabetic-induced experimental animal for 8 hours. R+C and R+C+C' are treated and pre-meal glucose (FBG) (6a) and post-prandial blood glucose (PBG) (6b) were measured at intervals of 2, 4, and 6 weeks.
7 is a result of measuring the anticoagulant activity (7a) and anti-spread platelet aggregation inhibitory ability (7b) of blood of raspberry extract.
8 is a normal diet (ND), high fat diet (HFD) and HFD+R, HFD+C, HFD+R+C, and HFD+R in a metabolic syndrome prevention experiment using highly obese inducing animals (C57BL/6N mice). This figure shows the results of carotoid intima medica thickness (CIMT; carotoid intima medica thickness) (8b) with the change of atherosclerosis index (AI; 8a) and arterial (Aorta) H&E staining when +C+C' was treated.

Hereinafter, the present invention will be described in detail.

The present invention is raspberry ( Rubus crataegifolius ) extract, hawthorn ( Crataegus pinnatifida ) extract and cinnamon ( Cinnamomum cassia ) provides a pharmaceutical composition for preventing or treating metabolic syndrome disease containing a mixture of the extract as an active ingredient.

The raspberries used as an active ingredient in the composition of the present invention are deciduous broad-leaved shrubs belonging to the genus Rubus of the Rosaceae family, and are distributed in China, Russia, and Japan, including Korea. The leaves are ovate or oval and are often divided into 3~5 pieces. The flowers are white bisexual flowers and bloom in June, and the fruits are round and ripe in July-August with a dark red color. Since ancient times, fruit has been harvested and used for edible or medicinal purposes, and sulfation and immunity enhancing effects have been reported. Raspberries contain a large amount of phenolic compounds such as flavotoids, tannins, and phenolic acids, as well as anthocyanin, a water-soluble pigment, and triterpenes that have various pharmacological actions. It is known that Ryu exists. The raspberry in the present invention is Rubus crataegifolius .

The Raspberry ( Rubus cratagegifolius ) refers to the green fruit that the raspberry flowers bear, which is preferably obtained from April to September, more preferably from May to August, and most preferably from May to July. However, it is not limited thereto.

The raspberry extract may be prepared by a manufacturing method comprising the following steps.

1) extracting the raspberries by adding an extraction solvent;

2) filtering the extract of step 1); And

3) A step of drying after concentrating the filtered extract of step 2) under reduced pressure.

In the above method, the raspberries of step 1) can be used without limitation, such as those grown or commercially available.

In the above method, the extraction solvent in step 1) may be water, alcohol, or a mixture thereof. The alcohol may be a C ₁ to C ₂ lower alcohol, and specifically, the lower alcohol may be methanol or ethanol.

The extraction method of step 1) may be shaking extraction, Soxhlet extraction, or reflux extraction. In this case, the extraction temperature may be 40 to 80°C, specifically 50 to 70°C, but is not limited thereto. In addition, the extraction solvent may be added 1 to 20 times the weight of the raspberry used for extraction, may be 1 to 10 times, or may be added 5 to 15 times, but is not limited thereto. In addition, the extraction time may be 1 to 60 hours, 4 to 58 hours, 10 to 48 hours, or 15 to 30 hours, but is not limited thereto. In addition, the number of extractions may be 1 to 5 times, may be 3 to 5 times, and, for example, it is preferable to repeatedly extract 3 times, but is not limited thereto. The content of the active ingredient in the raspberry extract may be insignificant because extraction is not sufficiently performed in a range outside the conditions of the extraction temperature, extraction time, or number of extractions. Alternatively, the extraction yield may no longer increase, and thus the efficiency of the extraction operation may decrease.

In addition, in one aspect of the invention, in the extraction of step 1), ethanol may be 10% to 90% ethanol, 20% to 80% ethanol, or 30% to 70% ethanol, and the extraction time is 2 To 10 hours, 3 to 9 hours, 4 to 8 hours, or 5 to 7 hours.

The vacuum concentration in step 3) may be performed using a vacuum vacuum concentrator or a vacuum rotary evaporator. In addition, the drying may be reduced pressure drying, vacuum drying, boiling drying, spray drying or freeze drying, preferably spray drying or freeze drying.

In addition, the extract further provides a pharmaceutical composition for the prevention or treatment of metabolic syndrome diseases, containing a fraction of the organic solvent prepared by extracting with an organic solvent as an active ingredient. The organic solvent is preferably selected from the group consisting of n-hexane, methylene chloride, chloroform, ethyl acetate, dichloromethane, and butanol, but is not limited thereto.

On the other hand, the hawthorn used as an active ingredient in the composition of the present invention is also called agawi and is a red hawthorn fruit belonging to the Rosaceae family. The appearance is round and red, with white spots. Sansa is phenolic substances such as quercetin, hyricetin, gentisic acid, caffeic acid, catechin, and chlorogenic acid, and fluoroglucinol. Antioxidant substances such as (phloroglucinol), protocatechuic acid, and pyrogallol, amino acids such as glutamic acid, asparagine, and proline, and minerals such as potassium, calcium, and magnesium Contains. Since ancient times, it has been widely used for medicinal and edible use, and has been reported to have physiological activities such as antibacterial activity, antioxidant activity, inhibition of differentiation of fat progenitor cells, improvement of lipid concentration in the body, antihyperlipidemia, vascular relaxation effect, and liver damage prevention. The hawthorn in the present invention is Crataegus pinnatifida .

The hawthorn extract may be prepared by a manufacturing method comprising the following steps.

1) extracting by adding an extraction solvent to the hawthorn;

2) filtering the extract of step 1); And

3) A step of drying after concentrating the filtered extract of step 2) under reduced pressure.

In the above method, the hawthorn of step 1) can be used without limitation, such as cultivated ones or commercially available ones.

In the above method, the extraction solvent in step 1) may be water, alcohol, or a mixture thereof. The alcohol may be a C ₁ to C ₂ lower alcohol, and specifically, the lower alcohol may be methanol or ethanol.

The extraction method of step 1) may be shaking extraction, Soxhlet extraction, or reflux extraction. In this case, the extraction temperature may be 40 to 80°C, specifically 50 to 70°C, but is not limited thereto. In addition, the extraction solvent may be added 1 to 20 times the weight of the hawthorn used for extraction, may be 1 to 10 times, or may be added 5 to 15 times, but is not limited thereto. In addition, the extraction time may be 1 to 60 hours, 4 to 58 hours, 10 to 48 hours, or 15 to 30 hours, but is not limited thereto. In addition, the number of extractions may be 1 to 5 times, may be 3 to 5 times, and, for example, it is preferable to repeatedly extract 3 times, but is not limited thereto. In a range outside the conditions of the extraction temperature, extraction time, or number of extractions, extraction may not be sufficiently performed, so that the content of the active ingredient in the hawthorn extract may be insignificant. Alternatively, the extraction yield may no longer increase, and thus the efficiency of the extraction operation may decrease.

In addition, in one aspect of the invention, in the extraction of step 1), ethanol may be 10% to 90% ethanol, 20% to 80% ethanol, or 30% to 70% ethanol, and the extraction time is 2 To 10 hours, 3 to 9 hours, 4 to 8 hours, or 5 to 7 hours.

The vacuum concentration in step 3) may be performed using a vacuum vacuum concentrator or a vacuum rotary evaporator. In addition, the drying may be reduced pressure drying, vacuum drying, boiling drying, spray drying or freeze drying, preferably spray drying or freeze drying.

In addition, the extract further provides a pharmaceutical composition for the prevention or treatment of metabolic syndrome diseases, containing a fraction of the organic solvent prepared by extracting with an organic solvent as an active ingredient. The organic solvent is preferably selected from the group consisting of n-hexane, methylene chloride, chloroform, ethyl acetate, dichloromethane, and butanol, but is not limited thereto.

On the other hand, cinnamon, which is used as an active ingredient in the composition of the present invention, is a bark of an arboreous tree of the camphor family, and grows wildly in southern China or Vietnam. The main ingredients of cinnamon are essential oils called cinnamon oil (cinnamon aldehyde 75-90%, cinnamyl aldehyde, etc.), cinnamic aldehyde, camphene cineol, linalool, eugenol, It contains ingredients such as sugar, fat, phosphorus, iron, and vitamins A, B1, and B2. Cinnamon has been used in various ways such as antiperspirant, antipyretic, pain reliever, as well as flavoring for food products. In addition, it has been reported that it has an antiseptic effect that promotes intestinal peristalsis and inhibits abnormal fermentation in the intestine. Cinnamon in the present invention is Cinnamonium cassia Blume.

The cinnamon extract may be prepared by a manufacturing method comprising the following steps.

1) extracting cinnamon by adding an extraction solvent;

2) filtering the extract of step 1); And

3) A step of drying after concentrating the filtered extract of step 2) under reduced pressure.

In the above method, the cinnamon of step 1) can be used without limitation, such as grown or commercially available.

In the above method, the extraction solvent in step 1) may be water, alcohol, or a mixture thereof. The alcohol may be a C ₁ to C ₂ lower alcohol, and specifically, the lower alcohol may be methanol or ethanol.

The extraction method of step 1) may be shaking extraction, Soxhlet extraction, or reflux extraction. In this case, the extraction temperature may be 40 to 80°C, specifically 50 to 70°C, but is not limited thereto. In addition, the extraction solvent may be added 1 to 20 times the weight of cinnamon used for extraction, may be 1 to 10 times, or may be added 5 to 15 times, but is not limited thereto. In addition, the extraction time may be 1 to 60 hours, 4 to 58 hours, 10 to 48 hours, or 15 to 30 hours, but is not limited thereto. In addition, the number of extractions may be 1 to 5 times, may be 3 to 5 times, and, for example, it is preferable to repeatedly extract 3 times, but is not limited thereto. The content of the active ingredient in the cinnamon extract may be insignificant because extraction is not sufficiently performed in a range outside the conditions of the extraction temperature, extraction time, or number of extractions. Alternatively, the extraction yield may no longer increase, and thus the efficiency of the extraction operation may decrease.

In addition, in one aspect of the invention, in the extraction of step 1), ethanol may be 10% to 90% ethanol, 20% to 80% ethanol, or 30% to 70% ethanol, and the extraction time is 2 To 10 hours, 3 to 9 hours, 4 to 8 hours, or 5 to 7 hours.

The vacuum concentration in step 3) may be performed using a vacuum vacuum concentrator or a vacuum rotary evaporator. In addition, the drying may be reduced pressure drying, vacuum drying, boiling drying, spray drying or freeze drying, preferably spray drying or freeze drying.

In addition, the extract further provides a pharmaceutical composition for the prevention or treatment of metabolic syndrome diseases, containing a fraction of the organic solvent prepared by extracting with an organic solvent as an active ingredient. The organic solvent is preferably selected from the group consisting of n-hexane, methylene chloride, chloroform, ethyl acetate, dichloromethane, and butanol, but is not limited thereto.

The present inventors studied a pharmaceutical composition for preventing or treating metabolic syndrome disease using natural plant extracts, while the complex active ingredients of raspberry, hawthorn and/or cinnamon extract significantly reduced the dietary amount of experimental animals and the secretion of leptin, a dietary hormone. And, by increasing adiponectin, it reduces body weight, fat and sugar tolerance, reduces triglycerides, cholesterol and LDL-cholesterol, increases HDL-cholesterol, decreases ALP, AST, and protects liver health, especially for blood coagulation and aggregation. Prevents metabolic syndrome, which can cause cerebrovascular disease and heart attack by promoting blood circulation by preventing and preventing metabolic syndrome, which can cause cerebrovascular disease and heart attack by showing remarkable obesity, fatty liver suppression, abnormal lipid suppression and blood circulation, carotid artery protection, and liver protection effect than a single extract. It was found that it can be usefully used for treatment.

In the composition of the present invention, the mixture of raspberry extract, hawthorn extract and cinnamon extract may be prepared by adding an extraction solvent to each dried product of raspberry, hawthorn and cinnamon to prepare a single extract, and mixing the single extract. Specifically, the mixture may be a mixture of raspberry extract, hawthorn extract, and cinnamon extract in a weight ratio of 1: 0.1-3: 0.1-3, or 1: 0.5-2.5: 0.5-2.5 in a weight ratio, or 1: 1 It may be mixed in a weight ratio of -2.5: 1-2.5, or a weight ratio of 1: 1.5-2.5: 1.5-2.5. For example, the mixture may be a mixture of raspberry extract, hawthorn extract, and cinnamon extract in a weight ratio of 1: 1-2: 1-2. In addition, the composition may be a composition comprising an extract obtained by first mixing two or three dried products of raspberries, hawthorn and cinnamon dried products at the weight ratio described above, and then extracting the mixture.

In one aspect of the present invention, the metabolic syndrome disease is not limited as including conditions, symptoms, or diseases included in commonly known metabolic diseases, but, for example, the metabolic syndrome disease is obesity, liver disease, Fatty liver, dyslipidemia, blood circulation disorder, thrombosis, arteriosclerosis, hypertension, hyperlipidemia or type 2 diabetes.

In another aspect of the present invention, the composition is for preventing or treating metabolic syndrome disease through weight loss, body fat reduction, total cholesterol reduction, total glyceride reduction, HDL-C elevation or LDL-C reduction, wherein the metabolic syndrome disease May be, for example, obesity, or dyslipidemia.

In another aspect of the present invention, the composition is a reduction in liver AST (aspartate aminotransferase) level, liver ALT (alanine aminotransferase) level, liver lipid reduction, liver tissue fat cell size and number reduction, liver FAS ( It is to prevent or treat metabolic syndrome disease by reducing the enzyme activity of fatty acid synthase), reducing the enzyme activity of HMG-CoAR (3-hydroxy-3-methyl-glutaryl-CoA reductase) in the liver, or increasing the activity of beta-oxidation-related CPT. , Wherein the metabolic syndrome disease may be, for example, liver disease or fatty liver, wherein the liver disease is a condition, symptom or disease in the liver that can be prevented, treated or ameliorated from the above-described effects.

In another aspect of the present invention, the composition is for preventing or treating a metabolic syndrome disease by controlling blood sugar or improving glucose tolerance, wherein the metabolic syndrome disease may be diabetes or impaired glucose tolerance, for example, type 2 diabetes.

In another aspect of the present invention, the composition is for preventing or treating metabolic syndrome diseases by inhibiting blood coagulation, inhibiting platelet aggregation, or inhibiting arteriosclerosis, wherein the metabolic syndrome diseases include, for example, blood circulation disorders, thrombosis, arteriosclerosis. It may be a hauling disorder caused by.

In another aspect of the present invention, the composition may be to prevent or treat arteriosclerosis, hypertension, or hyperlipidemia from any one of the above-described effects.

In a specific embodiment of the present invention, in order to confirm the preventive effect of single or complex extracts against diseases of metabolic syndrome, a single or complex extract was supplied together for 8 weeks to mice inducing obesity by a high fat diet. As a result, compared to the control group (HFD), the amount of diet was significantly decreased in the experimental group (HFD+R,C,R+C+R+C+C') (Table 1), and the secretion of the dietary hormone leptin was significantly reduced, It was confirmed that adiponectin was remarkably increased, so that body weight and body fat weight were significantly reduced (see FIG. 1), and liver weight and levels of liver enzymes AST and ALT were also significantly decreased (see FIG. 2). As shown in Fig. 1, weight loss was reduced by individual raspberry hawthorn extract (reduced 7.4% and 12.74), but the reduction effect was remarkably reduced when raspberry + hawthorn (16.2%) and raspberry + hawthorn + cinnamon (18.24%) were treated. It can be seen that it increases significantly.

In order to examine plasma lipid concentration and liver tissue, blood was sampled after the administration period of single or complex extracts, and plasma enzymes (AST, ALT) and lipid levels (Triglyceride, total cholesterol, LDL-cholesterol, HDL-cholesterol) were analyzed. Triglyceride and total cholesterol concentrations were analyzed, and changes in hepatocytes were analyzed morphologically (FIGS. 2, 3 and 5). As a result, in the study on the prevention of metabolic syndrome by highly obese-inducing animals, the liver weight of the experimental group (HFD+R,C,RC,RCC' was decreased compared to the control group (HFD) (Fig. It can be seen that the levels of AST/Figs. 2b and 2c) also decreased, especially when the combined treatment of raspberry + hawthorn extract and raspberry + hawthorn + cinnamon extract, liver levels decreased by 79.1%, AST (33.6% and 51.1%, respectively). ) It can be seen that it reduces and synergisticly reduces liver damage caused by HFD (FIGS. 2B and 2C ).

As shown in Figure 3, the R and C extract treatment showed a decrease in TC, TG and LDL levels, and in the R+C and R+C+C' extracts, respectively, TC (39.3%, 41.6%), TG (40.6%, 50.6%) and LDL (34.9%, 36.4%) decreased, so the R+C complex extract and the R+C+C' complex extract showed a synergistic healing effect, and HDL increased by 42.5% and 45.0%, respectively. As it shows, it can be an excellent material for improving abnormal lipid metabolism.

In addition, as a result of measuring liver enzyme activity using the method of Hulcher and Oleson (1973) to investigate liver lipid-forming enzymes, the activity of enzymes involved in liver lipid metabolism (FAS, HMG-CoAR) in the experimental group compared to the control group (HFD). This decreased and the activity of CPT involved in beta-oxidation increased (FIG. 4).

In Figure 5, the presence and reduction of fat droplets in liver tissue was measured by H&E Staining and Oil Red O Staining.

In addition, as shown in Figure 6, after induction of diabetes in obesity-inducing animals, single or complex (R, C, RC, RCC' extracts were treated to test for blood glucose load, and anti-diabetic evaluation analysis showed that both FBG and PBG were experimental groups ( It can be seen that R, C, RC, RCC' had lower blood glucose levels than the control (HFD), and the glucose tolerance was remarkably overcome by the complex extract (see FIG. 6).

In addition, in order to investigate the anticoagulant activity, as a result of measuring activated Partial Thromboplastin Time (aPTT), Prothrombin time (PT) and thrombin time (TT), raspberry extract inhibited blood coagulation. (See Fig. 7a). In addition, the antiplatelet inhibitory ability was confirmed at various concentrations in the platelet aggregation inhibitory inhibitory activity (see Fig. 7b). On the other hand, it was confirmed that the anticoagulant and antiplatelet activity of hawthorn and cinnamon appeared very weak.

Figure 8 shows the arterial sclerosis index (artherosclerosis Index) and arterial H&E staining of the treated group when treated with Normal, HFD, HFD+SMV and R, C, R+C, R+C+C' in HFD-administered C57BL/6 mice. I got it.

Arteriosclerosis index (AI) was decreased in the experimental group (HFD+RFex) compared to the control group (HFD) (FIG. 8A). In addition, as a result of Aorta H&E staining, the intima thickness of aorta was similar to that of HFD treatment when treated with raspberries, so it could not alleviate arteriosclerosis, but when treated with hawthorn, hawthorn + raspberry, hawthorn + raspberry + cinnamon, the intima thickness decreased, leading to arteriosclerosis. You can see the effect of blocking. The thickness of the blood vessel's inner media is influenced by several factors. Cardiovascular risk factors such as age, sex, obesity, hypertension, hyperlipidemia, diabetes, and smoking are known to be closely related to the carotid intima thickness. From these facts, it can be seen that the single or complex extracts used in this study can be an important pharmaceutical composition for the prevention and treatment of blood vessels caused by metabolic diseases such as hypertension or hyperlipidemia, which relieves the carotid intima thickness.

In other words, when using a combination of raspberry and hawthorn extract used in this study, it prevents arteriosclerosis by reducing blood clotting, inhibiting platelet aggregation, and reducing the thickness of the blood vessel lining, which helps blood circulation, thereby facilitating blood circulation in the body, resulting in high blood pressure. , It can be seen that it can be an object of remarkably improved metabolic disease improvement and suppression as an important composition for blood vessel prophylaxis treatment by preventing abnormal lipid accumulation, etc.

Therefore, overall, the single or complex extract according to the present invention remarkably inhibits the expression of enzymes involved in lipid metabolism, reduces body weight, fat and sugar tolerance, and reduces triglyceride (TG) and cholesterol, and LDL-cholesterol. It can be seen that it can suppress cerebrovascular disease and heart attack, since it has been confirmed that it increases HDL-cholesterol, inhibits fatty liver, promotes blood circulation, and normalizes the inner medial thickness of blood vessels. It can be usefully used as a composition for preventing or treating metabolic syndrome showing a significant synergistic effect.

The pharmaceutical composition according to the present invention may contain 10 to 95% by weight of a mixture of the active ingredients of raspberry extract, hawthorn extract, and cinnamon extract based on the total weight of the composition. In addition, the pharmaceutical composition of the present invention may further include one or more active ingredients exhibiting the same or similar functions in addition to the active ingredients.

The pharmaceutical composition of the present invention may contain a carrier, a diluent, an excipient, or a mixture thereof commonly used in biological preparations. Any pharmaceutically acceptable carrier can be used as long as it is suitable for delivering the composition in vivo. Specifically, the carrier is Merck Index, 13th ed., Merck & Co. Inc., saline, sterile water, Ringer's solution, dextrose solution, maltodextrin solution, glycerol, ethanol, or a mixture thereof. In addition, conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added as needed.

When formulating the composition, diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants may be added.

The composition of the present invention may be formulated as an oral or parenteral formulation. Oral formulations may include solid formulations and liquid formulations. The solid preparation may be a tablet, a pill, a powder, a granule, a capsule or a troche, and the solid preparation may be prepared by adding at least one excipient to the composition. The excipient may be starch, calcium carbonate, sucrose, lactose, gelatin, or a mixture thereof. In addition, the solid preparation may contain a lubricant, and examples thereof include magnesium stearate and talc. On the other hand, the liquid formulation may be a suspension, an inner solution, an emulsion or a syrup. At this time, the liquid formulation may contain excipients such as wetting agents, sweetening agents, fragrances, and preservatives.

The parenteral preparation may include injections, suppositories, powders for respiratory inhalation, aerosols for sprays, powders and creams. The injection may include a sterilized aqueous solution, a non-aqueous solvent, a suspension solvent, an emulsion, and the like. At this time, as the non-aqueous solvent or suspension solvent, vegetable oils such as propylene glycol, polyethylene glycol, and olive oil, or injectable esters such as ethyl oleate may be used.

The composition of the present invention may be administered orally or parenterally according to a desired method. Parenteral administration may include intraperitoneal, rectal, subcutaneous, intravenous, intramuscular or intrathoracic injection.

The composition may be administered in a pharmaceutically effective amount. This may vary depending on the type of disease, the severity, the activity of the drug, the patient's sensitivity to the drug, the administration time, the administration route, the treatment period, and the drugs used at the same time. However, for a desirable effect, the amount of the active ingredient contained in the pharmaceutical composition according to the present invention may be 0.0001 to 1,000 mg/kg, specifically 0.001 to 500 mg/kg. The administration may be several times a day.

The composition of the present invention may be administered alone or in combination with other therapeutic agents. When administered in combination, administration may be sequential or simultaneous.

In addition, the present invention raspberry ( Rubus crataegifolius ) extract, hawthorn ( Crataegus pinnatifida ) extract and cinnamon ( Cinnamomum cassia ) provides a health functional food composition for preventing or improving metabolic syndrome disease containing a mixture of the extract as an active ingredient.

The raspberry, hawthorn and cinnamon extract may be the same as described in the pharmaceutical composition. For example, the extract _{may be extracted with water, C 1} to C ₂ lower alcohol or a mixture thereof. Specifically, the lower alcohol may be methanol or ethanol. More specifically, the lower alcohol may be 30 to 70% ethanol or 40 to 60% ethanol, and according to an embodiment, the lower alcohol may be 50% ethanol.

In addition, in one aspect of the invention, in the extraction of step 1), ethanol may be 10% to 90% ethanol, 20% to 80% ethanol, or 30% to 70% ethanol, and the extraction time is 2 To 10 hours, 3 to 9 hours, 4 to 8 hours, or 5 to 7 hours.

In the composition of the present invention, the mixture of raspberry extract, hawthorn extract, and cinnamon extract may be prepared by adding an extraction solvent to each dried product of raspberry, hawthorn and cinnamon to prepare a single extract, and mixing the single extract. Specifically, the mixture may be a mixture of raspberry extract, hawthorn extract, and cinnamon extract in a weight ratio of 1: 0.1-3: 0.1-3, or 1: 0.5-2.5: 0.5-2.5 in a weight ratio, or 1: 1 It may be mixed in a weight ratio of -2.5: 1-2.5, or a weight ratio of 1: 1.5-2.5: 1.5-2.5. For example, the mixture may be a mixture of raspberry extract, hawthorn extract, and cinnamon extract in a weight ratio of 1: 1-2: 1-2. In addition, the composition may be a composition comprising an extract obtained by first mixing two or three kinds of dried raspberries, hawthorn and cinnamon at the weight ratio described above, and then extracting the mixture.

The health functional food composition regulates the amount of diet and thus decreases the dietary hormone leptin and increases adiponectin, thereby reducing body weight, fat and sugar tolerance, reducing triglycerides, cholesterol and LDL-cholesterol, and HDL- Cerebral vascular disease by increasing cholesterol and reducing ALP and AST to protect liver health and promoting blood circulation by preventing blood coagulation and aggregation, and comprehensively suppresses obesity and fatty liver, suppresses abnormal lipids and blood circulation, protects the carotid artery, and protects the liver. And it may be usefully used in the prevention or improvement of metabolic syndrome disease that may cause a heart attack.

In a specific embodiment of the present invention, in order to confirm the preventive effect of single or complex extracts against diseases of metabolic syndrome, a single or complex extract was supplied together for 8 weeks to mice inducing obesity by a high fat diet. As a result, compared to the control group (HFD), body weight and body fat weight were decreased in the experimental group (HFD+R,C,R+C+R+C+C' (see Fig. 1)), and liver weight and liver enzymes AST, ALT It was confirmed that the numerical value of was also decreased (see Fig. 2.) As shown in Fig. 1, the weight loss showed a reduction effect individually (reduced 7.4% and 12.74), but raspberry + hawthorn (16.2%) and raspberry + It can be seen that the reduction effect increases when hawthorn + cinnamon (18.24%) is treated.

In order to examine plasma lipid concentration and liver tissue, blood was collected after the administration period of single or complex extracts, and plasma enzymes (AST, ALT) and lipid levels (Triglyceride, total cholesterol, LDL-cholesterol, HDL-cholesterol) were analyzed. Triglyceride and total cholesterol concentrations were analyzed, and changes in hepatocytes were analyzed morphologically (Figs. 2, 3, and 5). As a result, in the study on the prevention of metabolic syndrome by highly obese-inducing animals, the liver weight of the experimental group (HFD+R,C,RC,RCC' was decreased compared to the control group (HFD) (Fig. It can be seen that the levels of AST/Fig. 2b and 2c) also decreased, especially when the combined treatment of raspberry + hawthorn extract and raspberry + hawthorn + cinnamon extract, liver levels decreased by 79.1%, AST (33.5%, 51.0%, respectively) ), and it can be seen that synergistic reduction of liver damage caused by HFD (FIGS. 2b and 2c ).

As shown in Figure 3, the R and C extract treatment showed a decrease in TC, TG and LDL levels, and in the R+C and R+C+C' extracts, respectively, TC (39.3%, 41.6%), TG (40.6%, 50.6%) and LDL (33.2%, 40.5%) reduction effect, so R+C complex extract and R+C+C' complex extract show a synergistic healing effect, so it can be an excellent material for improving abnormal lipid metabolism. have.

In addition, as a result of measuring liver enzyme activity using the method of Hulcher and Oleson (1973) for the investigation of interlipid-forming enzyme, the activity of enzymes involved in liver lipid metabolism (FAS, HMG-CoAR) in the experimental group compared to the control group (HFD). It decreased and the activity of CPT involved in beta-oxidation increased (FIG. 4).

In Figure 5, the presence and reduction of fat droplets in liver tissue was measured by H&E Staining and Oil Red O Staining.

In addition, as shown in Figure 6, after induction of diabetes in obesity-inducing animals, single or complex (R, C, RC, RCC' extracts were treated to test for blood glucose load, and anti-diabetic evaluation analysis resulted in both FBG and PBG cases ( It can be seen that R, C, RC, and RCC' had lower blood glucose levels than the control (HFD), and the glucose tolerance was overcome by the complex extract (see FIG. 6).

In addition, in order to investigate the anticoagulant activity, as a result of measuring activated Partial Thromboplastin Time (aPTT), Prothrombin time (PT) and thrombin time (TT), raspberry extract inhibited blood coagulation. (See Fig. 7a). In addition, the antiplatelet inhibitory ability was confirmed at various concentrations in the platelet aggregation inhibitory inhibitory activity (see Fig. 7b). On the other hand, the anticoagulant and antiplatelet imaging properties of hawthorn and cinnamon were found to be very weak.

Figure 8 shows the arterial sclerosis index (artherosclerosis Index) and arterial H&E staining of the treated group when treated with Normal, HFD, HFD+SMV and R, C, R+C, R+C+C' in HFD-administered C57BL/6 mice. I got it.

Arteriosclerosis index (AI) was decreased in the experimental group (HFD+RFex) compared to the control group (HFD) (FIG. 8A). In addition, as a result of Aorta H&E staining, the intima thickness of aorta was similar to that of HFD treatment when treated with raspberries, so it could not alleviate arteriosclerosis. However, when treated with hawthorn, hawthorn + raspberry, hawthorn + raspberry + cinnamon, the intima thickness decreased, resulting in arteriosclerosis. You can see the effect of blocking. The thickness of the blood vessel's inner media is influenced by several factors. Cardiovascular risk factors such as age, sex, obesity, hypertension, hyperlipidemia, diabetes, and smoking are known to be closely related to the carotid intima thickness. From these facts, it can be seen that the single or complex extracts used in this study can be an important pharmaceutical composition for the prevention and treatment of blood vessels caused by metabolic diseases such as hypertension or hyperlipidemia, which relieves the carotid intima thickness. Therefore, the single or complex extract according to the present invention, the active ingredient inhibits the expression of enzymes involved in lipid metabolism, reduces body weight, fat and sugar tolerance, reduces triglycerides (TG) and cholesterol, LDL-cholesterol, and HDL- It has been confirmed that it increases cholesterol, inhibits fatty liver, promotes blood circulation, and normalizes the inner media thickness of blood vessels, so it can be seen that it can suppress cerebrovascular disease and heart attack, and the complex extract prevents metabolic syndrome. Or it can be usefully used as a health functional food composition for improvement.

"Health functional food" as used herein refers to a food manufactured using nutrients that are liable to be deficient in daily meals or raw materials or ingredients having useful functions for the human body, and refers to foods that help maintain human health, but is not limited thereto. It is not used in a sense that includes all health foods in the usual sense.

The form and type of the health functional food is not particularly limited. Specifically, the health functional food may be in the form of tablets, capsules, powders, granules, liquids, and pills. The health functional food may contain various flavoring agents, sweetening agents, or natural carbohydrates as additional ingredients. The sweetener may be a natural or synthetic sweetener, and examples of the natural sweetener include taumatin and stevia extract. Meanwhile, examples of synthetic sweeteners include saccharin and aspartame. In addition, the natural carbohydrates may be monosaccharides, disaccharides, polysaccharides, oligosaccharides and sugar alcohols.

In addition to the above-described additional ingredients, the health functional food of the present invention includes nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pexane and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, and preservatives. , Glycerin, alcohol, and the like may be further included. These ingredients can be used independently or in combination. The ratio of the additive may be selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

The [mixture of raspberry extract and hawthorn extract] or [mixture of raspberry extract, hawthorn extract, and cinnamon extract] of the present invention may be added to food as it is or used with other foods or food ingredients. At this time, the content of the added active ingredient may be determined according to the purpose. In general, the content of the health functional food may be 0.01 to 90 parts by weight of the total food weight. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of health control, the amount may be below the above range, and if there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

Hereinafter, the present invention will be described in detail by the following examples and experimental examples.

However, the following examples and experimental examples are merely illustrative of the present invention, and the contents of the present invention are not limited by the following examples and experimental examples.

<Example 1> Raspberries ( Rubus crataegifolius ) Preparation of extract

10 times 50% ethanol was added to the raspberry sample, followed by extraction at 60°C for 4 hours. The extract was filtered using filter paper, and the filtrate was concentrated under reduced pressure and dried to obtain a final raspberry extract.

<Example 2> Sansa ( Crataegus pinnatifida ) Preparation of extract

After removing sugar from the fruit portion of the hawthorn, 10 times the mass of the sample was added 70% ethanol, followed by extraction at 60°C for 6 hours. The extract was filtered using filter paper, and the filtrate was concentrated under reduced pressure and dried to obtain a final hawthorn extract.

< Example 3> Cinnamon ( Cinnamomum cassia Blume ) Preparation of extract

Cinnamon was extracted at 60° C. by adding distilled water 10 times, and the extract was filtered using filter paper, concentrated and dried to obtain a final cinnamon extract.

<Experimental Example 1> Preparation of obesity induction mouse model by high fat diet

<1-1> Study on the prevention of metabolic syndrome by obesity-inducing animals

Experimental animals were provided with C57BL/6N male mice from central laboratory animals (Seoul, Korea). This animal is allowed to freely consume solid feed and water to adapt to the breeding environment for 7 days, maintains a temperature of 22℃ and a relative humidity of 50 ±10%, and separates the day and night every 12 hours. I did. The mice were divided into 6 groups according to the egg mass method into a normal group (ND, normal diet), a control group (HFD, high fat diet), and an experimental group (HFD+R, HFD+C, HFD+RC and HFD+RCC'). Diet was supplied for 8 weeks using a high-fat diet (HFD, containing 60 kcal% fat) and a normal diet (ND, containing 10 kcal% fat) purchased from a central laboratory animal (Seoul, Korea). In addition, the experimental group was orally administered the extract of unripe bokbunja for 8 weeks along with the high fat diet.

All experimental animals were conducted by the approved animal management system and use committee, and were performed according to the institutional guidelines of the Korea Research Institute of Bioscience and Biotechnology.

Normal group (ND): A diet (Normal diet, 10% fat) was given for 8 weeks.

Control (HFD): A high fat diet (60% fat) was given for 8 weeks.

Experimental group (HFD+R, C, RC, RCC'): High fat diet (60% fat) and raspberry, hawthorn, raspberry + hawthorn, raspberry + hawthorn + cinnamon extract were administered orally for 8 weeks.

As a result, as a result of the study on the prevention of metabolic syndrome by obesity-inducing animals, the amount of diet significantly decreased in the experimental group (HFD+R, HFD+C, HFD+RC, HFD+RCC') compared to the control group (HFD) (Table 1). (HFD; 17.94g/week, SMV; 16.64g/week, HFD+RC; 15.73g/week, HFD+RCC'; 15.46g/week) The amount of leptin, a dietary hormone, significantly decreased, and combined with RC and RCC' treatment. It was confirmed that the effect of reducing the dietary hormone leptin of the extract was higher than that of treatment with a single extract (FIG. 1D). In addition, since the amount of adiponectin was significantly increased, it was confirmed that the adiponectin increasing effect was increased when the complex extract was treated than when the single extract was treated (Fig. 1d). In addition, as shown in FIGS. 1A to 1C, body weight and body fat weight were decreased in the experimental group. In addition, it was found that the weight of the liver and the levels of the enzymes AST and ALT in the liver were also reduced (FIGS. 2A to 2C ).

Normal group (ND) Control (HFD) Simvastin (SMV) High Fat Diet + Raspberry (HFD+R) High Fat Diet + Sansa (HFD+C) High fat diet + raspberries + hawthorn (HFD + RC) High fat diet + raspberries + hawthorn + cinnamon (HFD+RCC') Weight(g) 29.28±5.8** 51.93±2.19 48.85±5.76 48.48±1.58* 45.31±6.22* 44.3±5.7* 44.68±3.07** Dietary intake (g/week) 16.16±0.8* 17.94±0.61 16.64±0.42* 16.14±0.56 16.35±0.38 15.73±0.67 15.46±0.56 Dietary intake efficiency ratio 0.021 0.096 0.063 0.076 0.053 0.053 0.061 Total fat mass (g) 1.06±0.14** 4.73±0.35 4.26±0.39* 3.76±0.52* 3.71±0.3** 3.64±0.24** 3.69±0.25* Kidney membrane (g) 0.32±0.11** 1.41±0.2 1.37±0.23 1.13±0.12* 1.06±0.12** 0.96±0.16** 1.04±0.14* Abdominal fat (g) 0.21±0.09** 1.81±0.17 1.51±0.2* 1.39±0.26* 1.35±0.1** 1.39±0.08** 1.37±0.23** Epididymis fat (g) 0.49±0.12** 1.50±0.18 1.37±0.16 1.24±0.28* 1.28±0.19* 1.27±0.11* 1.23±0.16* Liver weight (g) 0.93±0.06 2.41±0.19 1.61±0.34 1.86±0.2 1.70±0.39 1.63±0.31 1.52±0.32

Dietary intake efficiency ratio = weight gain (g/day)/diet intake (g/day)

Statistical differences were analyzed by an independent t-test, and values were expressed as mean±standard deviation (n=7). The results were compared with the high fat diet (HFD) group using the student's t-test.

(** is p<0.005, * is p<0.05)

<1-2> Study on the treatment of metabolic syndrome by obesity-inducing animals

Experimental animals were provided with C57BL/6N male mice from central laboratory animals (Seoul, Korea). The animals are allowed to freely consume solid feed and water to adapt to the breeding environment for 7 days, maintain a temperature of 22°C and a relative humidity of 50 ±10%, and separate the day and night every 12 hours. I did. After inducing obesity for 8 weeks using a high-fat diet (HFD, containing 60 Kcal% fat) supplied from a central laboratory animal (Seoul, Korea), control (HFD, high-fat diet), experimental group (HFD+ treatment) according to the egg mass method Group). Then, the control group was fed a high-fat diet for 5 weeks, and the experimental group was orally administered R, C, RC, and RCC' extracts for 5 weeks along with the high-fat diet. All experimental animals were conducted by the approved animal management system and use committee, and were performed according to the institutional guidelines of the Korea Research Institute of Bioscience and Biotechnology.

<Experimental Example 2> Plasma lipid concentration and liver tissue examination

After the end of the single or mixed extract administration period, blood was collected using cardiac puncture, and plasma enzymes (AST, ALT) and lipid levels (Triglyceride, total cholesterol, LDL-cholesterol, HDL-cholesterol) were analyzed. Plasma was stored at -70℃ by centrifuging the blood (10.000 xg, 5 min, 4℃), and the levels of ALT, AST, TG, TC, LDL-C and HDL-C in plasma were automatically determined by KIRBB's animal laboratory. It was measured using a biochemical analyzer.

Interlipid extraction was used by modifying Folch's method (1957). The frozen liver tissue was homogenized using 0.9% NaCl solution, and CM solution (Chloroform-methanol, 1:2, v/v) was added, then the mixture was well mixed and centrifuged (2000xg, 20 minutes) to separate the supernatant. Filtered, and the separated chloroform layer was used for analysis. The concentrations of triglyceride and total cholesterol in the liver were analyzed using an enzyme assay kit (Asan Pharmaceutical Co. Ltd, Seoul, Korea).

<Experimental Example 3> Morphological analysis of liver

For morphological analysis of hepatocytes, H&E (Hematoxilin & Eosin) and Oil Red O staining were used. After fixing liver tissue in diluted 10% formaldehyde solution, dehydration with ethanol, embedding in paraffin and sectioning, hematoxylin-eosin (H&E) or Oil Red O Stained with, and observed using an optical microscope.

As a result, in a study on the prevention of metabolic syndrome by obesity-inducing animals, as shown in the figure, liver lipids (TG and TC) were decreased in the experimental group (HFD) compared to the control group (HFD), and the size and number of fat cells were also observed in the histological observation. Decreased (Fig. 5).

In addition, as a result of a study on the treatment of metabolic syndrome by obesity-inducing animals, plasma and liver lipid (TC and TG) levels were decreased in the experimental group (HFD) compared to the control group (HFD). Decreased (Fig. 3).

In addition, the arteriosclerosis index (AI) was decreased compared to the control group (FIG. 8).

<Experimental Example 4> Measurement of activity for investigation of interlipid-forming enzyme

Liver enzyme analysis was used by modifying the method of Hulcher and Oleson (1973). Use 0.1 mol/L triethanolamine, 0.02 mol/L ethylene diamine tetra-acetic acid, 2 mol/L dithiothreitol, pH 7.0 to homogenize and centrifuge ( 600xg, 10min) to remove cell debris, and the supernatant was centrifuged again (10,000xg, followed by 12,000 xg, 20 min, 4 °C) and then removed to obtain mitochondria. Then, the cytosol supernatant was separated through two ultracentrifugation (100,000xg, 60min, 4°C). The mitochondria and microsome pellets were re-dissolved in 800 µl of a homogenized solution, and bovine serum albumin was taken as a standard using the Bradford method, and proteins were quantified. FAS activity was measured by a part of the microsome, and according to Nepokroeff Lakshmanan's method (1975), by monitoring the oxidation of NADPH dependent on malonyl-CoA at 340 nm, the oxidized NADPH was measured at nmol/min/mg. Expressed in protein. CPT activity was calculated by mitochondrial fractionation according to Markwell's method (Markwell et al., 1973), and the result was expressed as a protein of nmol/min/mg. The activity of 3-hydroxy-3-methylglutaryl-coenzyme reductase (HMG-CoAR) of microsomes was performed using a modified Hulcher and Oleson's method (1973), and CoA-SH nmol It was expressed as a protein of /min/mg.

As a result, in a study on the prevention of metabolic syndrome by obesity-inducing animals, the activity of liver lipid metabolism-related enzymes (FAS, HMG-CoAR) decreased in the experimental group (HFD+R, C, RC, RCC') compared to the control group (HFD). And, beta-oxidation (beta-oxidation) involved CPT activity was increased (Fig. 4).

<Experimental Example 5> Analysis of antidiabetic evaluation of single, complex extracts of raspberries, hawthorn and cinnamon after induction of diabetes in obesity-inducing animals

Experimental animals were provided with C57BL/6N male mice from central laboratory animals (Seoul, Korea). The animals are allowed to freely consume solid feed and water to adapt to the breeding environment for 7 days, maintain a temperature of 22°C and a relative humidity of 50 ±10%, and separate the day and night every 12 hours. I did. After adaptation to the environment, the mice were divided into two groups: a normal group (ND, normal diet) and a control group (HFD, high fat diet) according to the egg mass method, and were supplied with diet. After supplying ND and HFD for 8 weeks, the glucose level of the blood of each mouse was measured, and the blood glucose level in the HFD-treated group was measured, and then a single or complex extract was treated to perform a blood glucose test.

Experiments were conducted for 0, 2, 4 and 6 weeks. After starving for 8 hours in the evening, the amount of glucose in the mouse blood was measured (FBG; fasting blood glucose) and glucose (1.0 g/kg) was administered orally. After 2 hours, the amount of glucose in the blood was measured (PBG; postprandial blood blucose level). All experimental animals were conducted by the approved animal management system and use committee, and were performed according to the institutional guidelines of the Korea Research Institute of Bioscience and Biotechnology.

Normal group (ND): 8 weeks + 6 weeks of diet (Nnrmal diet, 10% fat) was given.

Control (HFD): 8 weeks + 6 weeks of diet (High diet, 60% fat) was given.

Experimental groups (HFD+R, C, RC, RCC' diet (high fat diet, 60% fat) were given for 6 weeks, and R, C, RC, RCC' extracts were administered orally for 4 weeks. As a result, both FGB and PBG were administered. In the case of the experimental group (HFD+RFex), the glucose level in the blood was lower than that of the control group (HFD), and the glucose tolerance was overcome by a single or complex extract (FIG. 6).

<Experimental Example 6> Measurement of anticoagulant activity of raspberry extract powder

Human blood collection

In order to investigate the anticoagulant activity and the ability to inhibit platelet aggregation in Experimental Example 7, plasma was prepared from whole blood from volunteers who did not receive drug administration for the last 1 month, and immediately after blood collection, centrifuged at 4℃ for 5 minutes at 4℃ for 5 minutes. Was separated and stored in a frozen state (fresh frozen plasma), and used after thawing at room temperature if necessary.

In order to investigate the anticoagulant activity of the raspberry extract of the present invention, activated Partial Thromboplastin Time (aPTT), Prothrombin time (PT) and thrombin time (Thrombin Time, TT) were measured. Anticoagulant activity was evaluated by measuring blood clotting time using the previously reported Amelung coagulometer KC-1A (Japan), and TT, aPTT and endogenous blood clotting time PT exogenous blood clotting time.

<6-1> Thrombin time (TT) measurement

Thrombin time was mixed with 50 µl of 0.5U thrombin (Sigma Co., USA) at 37°C in a tube of Amelung coagulometer KC-1A (Japan), reacted for 2 minutes, added 100 µl of plasma, and then plasma. The time until it solidified was measured. Aspirin (Sigma Co., USA) was used as a control, and DMSO was used instead of the sample as a solvent control. The degree of anticoagulant activity was confirmed by measuring TT at various concentrations by the above method.

As a result, blood coagulation activity was confirmed (Table 2).

<6-2> Measurement of activated Partial Thromboplastin Time (aPTT)

For aPTT: activated Partial Thromboplastin Time (aPTT), 100 µl of plasma and 10 µl of sample extracts of various concentrations were added to the tube of Amelung coagulometer KC-1A (Japan), heated at 37°C for 3 minutes, and 50 µl of aPTT Reagent (Sigma, ALEXIN ^™ ) was added and incubated again at 37° C. for 3 minutes. Thereafter, 50 µl CaCl ₂ (35 mM) was added and the time until plasma coagulated was measured. DMSO was used instead of the sample as a solvent control.

As a result, raspberry extract inhibited blood coagulation (Table 2).

<6-3> Prothrombin time (PT) measurement

70 µl of prothrombin time with standard plasma (MD Pacific Co., China) and 10 µl of sample solution of various concentrations were added to the tube of Amelung coagulometer KC-1A (Japan), heated at 37°C for 3 minutes, and then 130 µl of PT The time until the reagent was added and the plasma coagulated was indicated. Aspirin (Sigma Co., USA) was used as a control, and DMSO was used instead of the sample as a solvent control.

As a result of measuring the degree of anticoagulant activity of raspberry extract of the present invention at various concentrations of the extract, it was confirmed that the raspberry extract of the present invention has anticoagulant activity (Table 2).

Sample Conc.
(mg/ml) Anti-thrombosis activity TT PT aPTT DMSO - 1.0 ±0.0 1.0 ±0.0 1.0 ±0.0 Aspirin 1.5 2.3 ±0.2 1.4 ±0.2 1.1 ±0.0 Unripe raspberry extract 5.0 ＞15 ＞15 ＞15 4.5 ＞15 ＞15 ＞15 4.0 ＞15 4.4 ±0.1 ＞15 3.5 ＞15 2.0 ±0.1 ＞15 3.0 1.3 ±0.0 1.5 ±0.0 ＞15 2.5 1.3 ±0.0 1.5 ±0.2 ＞15 1.25 1.2 ±0.4 1.1 ±0.3 1.1 ±0.0

As shown in Table 2, raspberry extract showed excellent blood coagulation effect by inhibiting the exogenous coagulation factor (PT), the endogenous coagulation factor (aPTT), and the common final coagulation factor (TT). On the other hand, the non-suri hawthorn extract showed a weak blood coagulation effect (DATA not shown).

<Experimental Example 7> Measurement of inhibitory activity against platelet aggregation inhibition of raspberry extract powder

Platelet aggregation inhibition activity: After mixing with 3.2% sodium citrate solution from healthy human whole blood in a ratio of 1:9 ml, mixed with unripe raspberry extract sample. Inject a mixed sample of blood and unripe raspberry extract into a cartridge (collagen-epinephrine), and use PFA-100 (Platelet adhesion & aggregation test) to determine the degree of inhibition of platelet aggregation inhibitory activity and platelet aggregation of unripe raspberry extract. Measurements were made at various concentrations.

As a result, it was confirmed that it has the ability to inhibit platelet aggregation (Fig. 7).

As shown in Figure 7, the raspberry extract was able to observe the platelet inhibitory ability by ADP. In addition, the effect of inhibiting platelet aggregation by collagen can be observed. As described above, it was proved that the unripe raspberry extract showed strong anticoagulant and antiplatelet aggregation effects.

Figure 8 shows the arterial sclerosis index (artherosclerosis Index) and arterial H&E staining of the treated group when treated with Normal, HFD, HFD+SMV and R, C, R+C, R+C+C' in HFD-administered C57BL/6 mice. I got it.

Arteriosclerosis index (AI) was decreased in the experimental group (HFD+RFex) compared to the control group (HFD) (FIG. 8A). In addition, as a result of Aorta H&E staining, the intima thickness of aorta was similar to that of HFD treatment when treated with raspberries, so it could not alleviate arteriosclerosis. However, when treated with hawthorn, hawthorn + raspberry, hawthorn + raspberry + cinnamon, the intima thickness decreased, resulting in arteriosclerosis. You can see the effect of blocking.

Hereinafter, a preparation example including the raspberry extract, hawthorn extract and cinnamon extract or fractions thereof of the present invention will be described, but the present invention is not intended to limit them, but is intended to be described in detail only.

<Production Example 1> Preparation of pharmaceutical formulation

<1-1> Preparation of powder

Mixture of raspberry extract, hawthorn extract and cinnamon extract 2 g

Lactose 1 g

The above ingredients were mixed and filled in an airtight cloth to prepare a powder.

<1-2> Preparation of tablets

Mixture of raspberry extract, hawthorn extract and cinnamon extract 100 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above ingredients, tablets were prepared by tableting according to a conventional tablet preparation method.

<1-3> Preparation of capsule formulation

Mixture of raspberry extract, hawthorn extract and cinnamon extract 100 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above ingredients, a gelatin capsule was filled according to a conventional capsule preparation method to prepare a capsule.

<1-4> Preparation of ring

Mixture of raspberry extract, hawthorn extract and cinnamon extract 1 g

Lactose 1.5 g

glycerin 1 g

Xylitol 0.5 g

After mixing the above components, it was prepared so as to be 4 g per pill according to a conventional method.

<1-5> Preparation of granules

Mixture of raspberry extract, hawthorn extract and cinnamon extract 150 mg

Soybean extract 50 mg

glucose 200 mg

Starch 600 mg

After mixing the above ingredients, 100 mg of 30% ethanol was added, dried at 60°C to form granules, and then filled into a cloth.

<Production Example 2> Preparation of health food

Mixture of raspberry extract, four extract and cinnamon extract 150 mg

Vitamin mixture Appropriate amount

Vitamin A acetate 70 ㎍

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

Vitamin B12 0.2 ㎍

Vitamin C 10 mg

Biotin 10 ㎍

Nicotinic acid amide 1.7 mg

Folic acid 50 ㎍

Calcium pantothenate 0.5 mg

Mineral mixture Appropriate amount

Ferrous sulfate 1.75 mg

Zinc oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium Phosphate Monobasic 15 mg

Dicalcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

The composition ratio of the vitamin and mineral mixture is relatively suitable for health food, but it may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. , To prepare granules, and can be used for preparing a health food composition according to a conventional method.

<Production Example 3> Preparation of food

<3-1> Preparation of flour food

0.5 to 5.0 parts by weight of a mixture of the raspberry extract, hawthorn extract, and cinnamon extract of the present invention was added to flour, and the mixture was used to prepare bread, cakes, cookies, crackers and noodles.

<3-2> Preparation of soup and gravy

0.1 to 5.0 parts by weight of a mixture of the raspberry extract, hawthorn extract, and cinnamon extract of the present invention were added to soups and broth to prepare health-promoting meat products, noodle soup, and broth.

<3-3> Preparation of ground beef

Ground beef for health promotion was prepared by adding 10 parts by weight of a mixture of raspberry extract, hawthorn extract and cinnamon extract of the present invention to ground beef.

<3-4> Manufacture of dairy products

5 to 10 parts by weight of a mixture of the raspberry extract, hawthorn extract, and cinnamon extract of the present invention were added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

<3-5> Manufacturing of Seonsik

Brown rice, barley, glutinous rice, and adlay were gelatinized and dried by a known method, and then roasted, and then prepared into a powder having a particle size of 60 mesh with a grinder. Black soybeans, black sesame seeds, and perilla seeds were also steamed and dried by a known method, and then roasted, and then prepared into a powder having a particle size of 60 mesh with a grinder. The mixture of the raspberry extract, hawthorn extract and cinnamon extract of the present invention was concentrated under reduced pressure in a vacuum concentrator, sprayed and dried with a hot air dryer, and the resulting dried product was pulverized with a grinder to a particle size of 60 mesh to obtain a dry powder. The grains, seeds and unripe raspberries of the present invention were prepared by doubling them in the following ratio. Grains (30 parts by weight of brown rice, 15 parts by weight of barley, 20 parts by weight of barley), seeds (7 parts by weight of perilla, 8 parts by weight of black beans, 7 parts by weight of black sesame), raspberry extract, hawthorn extract and cinnamon extract of the present invention Mixture (3 parts by weight), Ganoderma lucidum (0.5 parts by weight), Rehmannia (0.5 parts by weight).

<3-6> Preparation of chewing gum

Gum base 20%

Sugar 76.36-76.76%

Mixture of raspberry extract, hawthorn extract and cinnamon extract 0.24-0.64%

Fruit flavor One %

water 2 %

Using the above composition and content, a chewing gum was prepared using a conventional method.

<3-7> Preparation of candy

Sugar 50-60%

corn syrup 39.26-49.66%

Mixture of raspberry extract, hawthorn extract and cinnamon extract 0.24-0.64%

Orange scent 0.1%

Candy was prepared using a conventional method with the above composition and content.

Claims (19)

Raspberry ( Rubus cratagegifolius ) extract and hawthorn ( Crataegus) pinnatifida ) A pharmaceutical composition for the prevention or treatment of metabolic syndrome diseases containing a mixture of extracts as an active ingredient.
According to claim 1, The mixture is a mixture further comprising a cinnamon (Cinnamomum cassia ) extract, a pharmaceutical composition for preventing or treating metabolic syndrome disease.
The pharmaceutical composition for preventing or treating metabolic syndrome according to claim 1 or 2, wherein the extract is _{extracted with water, C 1} to C _{2 lower alcohol or a mixture thereof.}
The pharmaceutical composition of claim 3, wherein the lower alcohol is methanol or ethanol.
The pharmaceutical composition of claim 2, wherein the mixture is a mixture of raspberry extract, hawthorn extract and cinnamon extract in a weight ratio of 1: 0.1-3: 0.1-3.
The method of claim 1, wherein the composition is to prevent or treat metabolic syndrome disease through weight loss, body fat reduction, fatty liver reduction, total cholesterol reduction, total glyceride reduction, HDL-C elevation or LDL-C reduction. Ever composition.
The method of claim 1, wherein the composition decreases liver AST (aspartate aminotransferase) levels, decreases liver ALT (alanine aminotransferase) levels, decreases liver lipids, decreases the size and number of liver tissue fat cells, liver FAS (Fatty acid). synthase), reducing the enzyme activity of HMG-CoAR (3-hydroxy-3-methyl-glutaryl-CoA reductase) in the liver, or increasing the CPT activity involved in beta-oxidation to prevent or treat metabolic syndrome diseases, Pharmaceutical composition.
The pharmaceutical composition of claim 1, wherein the composition prevents or treats metabolic syndrome diseases through blood sugar control or improvement of glucose tolerance.
The pharmaceutical composition according to claim 1, wherein the composition is for preventing or treating metabolic syndrome diseases by inhibiting blood coagulation, inhibiting platelet aggregation, inhibiting arteriosclerosis due to a decrease in the thickness of the vascular inner media, or inhibiting hypertension.
The pharmaceutical composition according to claim 1, wherein the metabolic syndrome disease is obesity, liver disease, fatty liver, dyslipidemia, blood circulation disorder, thrombosis, arteriosclerosis, hypertension, hyperlipidemia or type 2 diabetes.
Raspberry ( Rubus crataegifolius ) extract and hawthorn ( Crataegus) pinnatifida ) Health functional food composition for preventing or improving metabolic syndrome diseases containing a mixture of extracts as an active ingredient.
The health functional food composition for preventing or improving metabolic syndrome diseases according to claim 11, wherein the mixture is a mixture further comprising cinnamon (Cinnamomum cassia) extract.
The health functional food composition of claim 11 or 12, wherein the extract is _{extracted with water, C 1} to C ₂ lower alcohol or a mixture thereof.
The health functional food composition according to claim 13, wherein the lower alcohol is methanol or ethanol.
The health functional food composition of claim 12, wherein the mixture is a mixture of raspberry extract, hawthorn extract and cinnamon extract in a weight ratio of 1: 0.1-3: 0.1-3.
The method of claim 11 or 12, wherein the composition prevents or improves metabolic syndrome disease through weight loss, body fat reduction, fatty liver reduction, total cholesterol reduction, total glyceride reduction, HDL-C elevation or LDL-C reduction. That, health functional food composition.
The method of claim 11 or 12, wherein the composition is a decrease in liver AST (aspartate aminotransferase) level, decrease in liver ALT (alanine aminotransferase) level, decrease in liver lipid, decrease in the size and number of liver tissue fat cells, and Prevent or ameliorate metabolic syndrome diseases by decreasing the enzyme activity of FAS (Fatty acid synthase), reducing the enzyme activity of HMG-CoAR (3-hydroxy-3-methyl-glutaryl-CoA reductase) in the liver, or increasing the CPT activity involved in beta-oxidation. That is, a health functional food composition.
The health functional food composition according to claim 11 or 12, wherein the composition prevents or improves metabolic syndrome diseases by controlling blood sugar or improving glucose tolerance.
The method of claim 11 or 12, wherein the composition is to prevent or improve metabolic syndrome disease by inhibiting blood coagulation, inhibiting platelet aggregation, inhibiting arteriosclerosis due to a decrease in the thickness of the blood vessel lining, or inhibiting hypertension. Food composition.

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