KR101346879B1 - Health food and Composition comprising hydroxybiphenyl derivatives for treating and preventing obesity, diabetes, fat liver, or hyperlipidemia - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the prevention and treatment of obesity, diabetes, fatty liver, or hyperlipidemia containing hydroxybiphenyl derivatives (Hydroxybiphenyl derivates) as an active ingredient and to obesity, diabetes, fatty liver or hyperlipidemia containing a hydroxybiphenyl derivative It relates to a dietary supplement for symptomatic improvement. The hydroxybiphenyl derivative according to the present invention is effective in the prevention of obesity, diabetes, fatty liver, or hyperlipidemia through weight loss, fat tissue reduction, hypoglycemia, inhibition of triglyceride accumulation in liver tissue, triglyceride reduction in liver, and blood cholesterol lowering effect. It can be usefully used as a therapeutic and dietary supplement.

Description

Health food and Composition comprising hydroxybiphenyl derivatives for treating and preventing obesity, diabetes, fat liver, or hyperlipidemia}

The present invention is a pharmaceutical composition for the prevention and treatment of obesity, diabetes, fatty liver or hyperlipidemia containing hydroxybiphenyl derivatives (Hydroxybiphenyl derivates) as an active ingredient and to improve the symptoms of obesity, diabetes, fatty liver or hyperlipidemia containing a hydroxybiphenyl derivative It relates to a dietary supplement for.

Obesity is a disease caused by the collapse of the human body's energy balance due to genetic, environmental, and mental factors. It is already recognized as a “disease” that needs to be cured worldwide. The number of patients continues to increase due to lifestyle changes and industrialization. The trend is. According to a report by the World Health Organization (WHO), approximately 1 billion people are overweight worldwide and 300 million are obese patients with a BMI of 30 kg / m ² or more. Obesity is rapidly increasing worldwide, and because obesity causes complications such as diabetes, hypertension, hyperlipidemia, heart disease, and cancer rather than risking obesity itself, the demand for obesity is steadily increasing and There is great potential for growth (Carmenrodrgue et al., NEJM . 341: 1097-1105, 1999). This trend is rapidly spreading in industrialized countries, where 5-10% of total medical expenses are used in the obesity field.However, if the market for obesity drugs is developed, there is a lack of effective products to meet the demand. Is expected to record high growth.

The causes of obesity include genetic factors, eating habits, lifestyle, and imbalances in energy metabolism. Treatments of obesity include diet therapy, exercise therapy, behavioral therapy, medication, orthodontics, and gastrectomy. Drug treatments include appetite suppressants, food absorption inhibitors, and heat production accelerators, and various drugs are used for weight loss. Among them, Roche's Xenical, which largely inhibits lipolytic enzymes, prevents the absorption of fat into the body. And Abbott's 'Reductil', which acts on the central nervous system and reduces appetite, is leading the market. However, recently, due to serious side effects, 'Reductil' has been withdrawn from the market, and 'Xenical' is also restricted in use because it causes gastrointestinal side effects such as fatty stools, gas production, and decreased absorption of fat-soluble vitamins. There is no clear treatment for obesity.

Recently, the term 'metaflammation' has emerged for inflammatory reactions caused by oversupply of nutrients or metabolites, and obesity has been termed as 'chronic and low-level inflammation'. Research on the relationship between obesity and the immune system is actively underway (Tilg H et al., Nat Rev Immunol 6: 772-783, 2006; Shoelson SE et al., Gastroenterology , 132: 2169, 2007; Hotamisligil GS et al., Nature 444: 860, 2007). Obesity also occurs through a series of processes involving the aggregation of adipocyte precursors and the differentiation of new adipocytes, resulting in the storage of triglycerides in the cytoplasm. At this time, AMP-activated protein kinase (AMPK) is an enzyme that plays an important role in maintaining the energy balance in the body by regulating carbohydrate metabolism and lipid metabolism in the cell depending on the energy state in the cell. In other words, activation of AMPK promotes pathways that increase energy production, such as glucose transport and fatty acid oxidation. If this pathway does not work, lipogenesis, protein synthesis and gluconeogensis lead to obesity (Apiradee S. et al. , Am J Physiol Endocrinol Metab 290: E925-932, 2006; Yun S. Lee et al., Diabetes 55: 2256-2264, 2006; S. Fogarty et al., 1804 (3): 581-91, 2010 Eun Ju Kim et al., Mol Pharmacol 72: 62-72, 2007). Thus, substances that act to inhibit the inflammatory response and AMPK activating materials are attracting attention as a new target of obesity treatment.

In addition, obesity-induced non-alcoholic fatty liver disease (NAFLD) refers to a wide range of diseases including fatty liver or simple fatty liver with inflammation in the liver regardless of alcohol consumption, fatty liver disease, liver fibrosis, liver cirrhosis, etc. It is associated with hyperlipidemia, type 2 diabetes mellitus, insulin resistance, and can be considered as a liver sign of metabolic disease (Bugianesi E et al., Diabetologia 48: 634-642, 2005; Mensink RP et al., Curr Opin Lipidol 19: 25-29, 2008). In order to treat NAFLD, it is reported that monotherapy is not enough and a combination therapy of therapeutic agents that can treat various metabolic diseases is required.

In the present invention, hydroxybiphenyl derivates known to exhibit antimicrobial activity (Wonjun Seo et al., Arch Pharm Res 9 (3): 127-130, 1986) showed that some compounds not only showed excellent weight loss and hypoglycemic effects, but also had therapeutic effects on fatty liver and hyperlipidemia, thereby treating obesity, diabetes, fatty liver, or hyperlipidemia. The present invention was completed by confirming that it can be usefully used as a dietary supplement for improving symptoms of diabetes, fatty liver or hyperlipidemia.

An object of the present invention is to provide a pharmaceutical composition for the prevention and treatment of obesity, diabetes, fatty liver or hyperlipidemia, containing a hydroxybiphenyl derivative known to have an antimicrobial action as an active ingredient.

In addition, an object of the present invention is to provide a health functional food for improving the symptoms of obesity, diabetes, fatty liver or hyperlipidemia containing hydroxybiphenyl derivative.

In order to achieve the above object, the present invention is a hydroxybiphenyl derivative compound 2-allyl-p-phenylphenol, 6-allyl-o, o'-biphenyl or 2,2'- diallyl-p, p ' -Provides a pharmaceutical composition for the prevention and treatment of obesity, diabetes, fatty liver or hyperlipidemia, which contains a biphenol compound as an active ingredient, and a dietary supplement for improving the symptoms of patients with obesity, diabetes, fatty liver or hyperlipidemia.

Hydroxybiphenyl derivatives are biphenyl-containing compounds having a hydroxy group and an allyl group, and include 2-allyl-p-phenylphenol, 6-allyl-o, o'-biphenyl, 2,2'-diallyl-p, p′-biphenol compounds are known to exhibit antimicrobial effects (Wonjun Seo et al., Arch Pharm Res 9 (3): 127-130, 1986).

Surprisingly, the hydroxybiphenyl derivatives of 2-allyl-p-phenylphenol, 6-allyl-o, o'-biphenyl or 2,2'-diallyl-p, p'-biphenol compounds are known as obesity, diabetes, Excellent effect on the prevention and treatment of fatty liver or hyperlipidemia.

The structures of the above-mentioned compounds are represented by the following Chemical Formulas 1-3.

The hydroxybiphenyl derivatives represented by Chemical Formulas 1 to 3 according to the present invention have been shown to play an important role in lipolysis by AMPK (AMP activated protein kinase) and ACC (Acetyl-CoA carboxylation) activation. Weight loss, adipose tissue reduction, hypoglycemic effect, hepatic fat accumulation, hepatic triglyceride reduction, and blood LDL (low density lipoprotein) reduction in obese mice induced by Indicates. In addition, mice fed with a high glucose diet also exhibited a hypoglycemic effect.

The hydroxybiphenyl derivatives of Chemical Formulas 1 to 3 according to the present invention have a therapeutic effect against obesity as well as obesity, and are useful as an agent for treating obesity, diabetes, fatty liver, or hyperlipidemia, and may be used for preventing and preventing disease progression. In addition, it is useful as a dietary supplement for improving the symptoms of obesity, diabetes, fatty liver or hyperlipidemia.

The pharmaceutical composition for the prevention and treatment of obesity, diabetes, fatty liver or hyperlipidemia according to the present invention can be used in the form of general pharmaceutical preparations, and can be used in the form of health functional food for the purpose of preventing and improving obesity, diabetes, fatty liver or hyperlipidemia. have.

The hydroxybiphenyl derivatives of Chemical Formulas 1 to 3 according to the present invention may be prepared by oral administration by mixing pharmaceutically acceptable conventional excipients, binders, disintegrants, lubricants, surfactants, cosolvents, sweeteners, and the like. can do. At this time, as an excipient, microcrystalline cellulose, lactose, corn starch, mannitol, low-substituted hydroxypropyl cellulose, magnesium metasilicate aluminate, colloidal silicon dioxide, etc. are preferable, and as a binder, hydroxypropyl methyl cellulose, povidone, hydride, etc. Roxypropyl cellulose is preferred, and disintegrant is preferably croscarmellose sodium, sodium starch glycolate, calcium carboxymethyl cellulose, crospovidone and the like, and the lubricant is preferably magnesium stearate, talc, sodium stearyl fumarate, or the like. Do. Surfactants include pharmaceutically acceptable polyoxyethylene sorbitan fatty acid esters, polyoxyethylene polyoxypropylene block copolymers, macrogol glycerol hydroxystearate and sodium lauryl sulfonate, which may be used alone or in combination of two or more thereof. Composites can be used. Co-solvents alone or in combination, such as propylene carbonate, propylene glycol, ethanol, transcutol (diethylene glycol monoethyl ether), glycofurol (Glycofurol, tetrahydrofurfuryl alcohol polyethylene glycol ether) and polyethylene glycols You can use Of these, propylene carbonate, propylene glycol and polyethylene glycols may be used alone or in combination of two or more thereof.

The present invention enables the preparation of pills, powders, granules, tablets, capsules, liquid formulations and the like as oral preparations, and the present invention further provides a pharmaceutically acceptable carrier in addition to the above-described active ingredients for administration. It is prepared by including one or more, the pharmaceutically acceptable carrier can be used by mixing one or more of the saline solution, sterile water, Ringer's solution, buffered saline solution, dextrose solution, maltodextrin solution, glycerol, ethanol and these components If necessary, other conventional additives such as antioxidants, buffers and bacteriostatic agents may be added. In addition, diluents, dispersants, surfactants, binders and lubricants may be additionally added to prepare injectable formulations such as aqueous solutions, suspensions, emulsions and the like.

In addition, the present invention may be prepared as a pouch, beverage or tea, when prepared in a liquid formulation, and may be prepared as a housewife raw material or added to other foods. At this time, food supplements that can be used food acceptable.

In addition, the dosage or dose of the hydroxybiphenyl derivative compound according to the present invention varies in the range depending on the weight, age, sex, health condition, diet, time of administration, administration method, excretion rate and severity of the patient. For example, the dosage of 0.01 mg / kg to 1000 mg / kg may be divided once to several times daily.

Accordingly, the present invention provides an obesity containing 2-allyl-p-phenylphenol, 6-allyl-o, o'-biphenyl or 2,2'-diallyl-p, p'-biphenol compound as an active ingredient, It can be usefully used as a pharmaceutical composition for treating and preventing diabetes, fatty liver or hyperlipidemia, and as a dietary supplement for improving obesity, diabetes, fatty liver or hyperlipidemia.

Figure 1 shows the effect of increasing the expression level of the active (phosphorylated form) protein of AMPK and ACC with respect to the compounds of formulas 1 to 3 in HepG ₂ cells.
Figure 2 shows the effect of inhibiting weight gain with respect to the compound of formula 1 to 3 in the obesity-induced mice providing a high fat diet.
Figure 3 provides a high fat diet showing the effect of reducing the adipose tissue with respect to the compound of formula 1 to 3 in the obesity-induced mice.
Figure 4 provides a high-fat diet showing a blood sugar improving effect on the compound of formula 1 to 3 in hyperglycemia induced mice.
Figure 5 shows the effect of inhibiting and improving the accumulation of fat globules in the liver tissues for the compounds of formulas 1 to 3 in mice induced by fatty liver due to high fat diet.
Figure 6 shows the triglyceride-lowering effect on the compound of formulas 1 to 3 in mice with elevated liver triglycerides due to high fat diet.
Figure 7 shows the blood LDL cholesterol-lowering effect on the compound of formula 1 to 3 in the mice high in the blood LDL cholesterol due to high fat diet.
Figure 8 shows a hypoglycemic effect on the compound of formula 1 to 3 in the hyperglycemia induced mice providing a high glycemic diet.

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited by the following examples.

< Example  1> AMPK  And ACC  Activation effect

The effect of the compounds according to the invention on the activation of AMPK and ACC, important proteins involved in fat metabolism was evaluated.

AMPK and ACC, respectively, phosphorylate to inhibit fat synthesis and regulate energy metabolism to consume the accumulated fat. HepG2 cells, a liver cancer cell line, were treated with 0.5 mM AICAR (aminoimidzole carboxamide ribonucleotide), which was identified as an AMPK activator, and compounds 1 to 3 at a concentration of 25 μM, followed by Western blot phosphorylation of AMPK and ACC. Check the degree to evaluate the degree of activation is shown in FIG.

As can be seen in Figure 1, the compounds according to the invention all increased the degree of phosphorylation of AMPK, ACC, suggests that they play an important role in lipolysis by activating AMPK and ACC.

< Example  2> On high paper  Effects in obese mice induced by

C57BL / 6 mice (male, 4 weeks old) were obtained from Orient Co., Ltd. for 6 weeks to provide high fat diet (60% fat calorie, Diet Research # D12492) and induced obesity (DIO mice, diet induced obese) mice, the normal control group was fed a standard diet (Standard chow, 13.5% fat calorie, Lab diet # 5001). After 6 weeks, divided into 5 test groups (n = 10 per group) according to body weight, and fed the normal diet to the normal control group (test group 1) mice and the obesity-inducing mice (test groups 2 to 5) during the entire test period. The high-fat diet was supplied and maintained in a dark: light cycle (12 hours: 12 hours intervals) and water was freely consumed. The diet, dose and administration method administered to the experimental group are shown in Table 1 below.

One) Weight measurement

C57BL / 6 mice, which induced obesity for 6 weeks in a high-fat diet, were separated into test groups as shown in Table 1, and then prepared in the form of a suspension as a suspension using 3% HPMC. Table 2 and FIG. In order to compensate for the placebo effect of 3% HPMC and the weight loss due to the administration stress, 3% HPMC was orally administered to the vehicle control group every day.

As shown in Table 2 and FIG. 2, the normal control group gained 7.5g in weight for 8 weeks, while the high-fat diet vehicle control group increased by 14.2 g, when providing a high-fat diet when providing a regular diet. It was confirmed that the weight gain was obvious when the high fat diet was provided in this test model.

When the compounds 1 to 3 are administered to the obese mouse model, the weight gain is significantly inhibited compared to the vehicle-administered group, suggesting that it can be used as a dietary supplement for preventing and treating obesity or improving obesity symptoms.

2) Fat tissue weight measurement

After completion of the compound administration for 8 weeks in the experiment 1), the adipose tissue (epididymal fat, retroperitoneal fat) of all administration groups were extracted and weighed, and is shown in FIG. 3.

In FIG. 3, epididymal fat and reperritoneal fat were measured, respectively, as compared to the control group with a normal diet, the control group provided with a high-fat diet was found to significantly increase the weight of adipose tissue. On the other hand, in all groups to which Compounds 1 to 3 were administered, adipose tissue weight was significantly reduced compared to vehicle control group.

Through this, it can be seen that the weight loss effect due to the administration of the compounds 1 to 3 compared to the vehicle administration group mainly due to the weight loss of adipose tissue, from these results Compound 1 to 3 by reducing the fat tissue to prevent and treat obesity or obesity It can be used as a dietary supplement for improving symptoms.

3) Blood glucose measurement

Obesity and diabetes are metabolic disorders and are highly correlated. Therefore, in order to determine the degree of improvement in blood sugar of the compound according to the present invention, fasting blood glucose was measured after completion of drug administration for 8 weeks, and is shown in FIG. 4.

As can be seen in Figure 4, as a result of administering five compounds, the compound according to the present invention showed a significant hypoglycemic effect compared to the vehicle control.

Therefore, 1 to 3 according to the present invention suggests that it can be used as a dietary supplement for preventing and treating diabetes or improving diabetic symptoms.

4) Liver tissue pictures

The degree of fat accumulation in liver tissue due to the high fat diet was checked, and the effect of the compound on the inhibition of fat accumulation in liver tissue was evaluated. After autopsy, 5 livers per group were extracted, tissue sections were prepared, and H & E staining was performed to observe the distribution of fat globules (FIG. 5). Fat diet was not observed in the liver of the normal diet group, but the vehicle control group fed the high fat diet showed a significant increase in the fat sphere. On the other hand, the group administered with Compounds 1 to 3 showed all excellent effects compared to the vehicle due to the effect of the drug (Table 3). These results suggest that the compound according to the present invention can be used as a dietary supplement for preventing and treating fatty liver or improving symptoms of fatty liver.

5) Liver Triglyceride ( TG ) Measure

The degree of fat accumulation in liver tissue due to the high fat diet was evaluated, and the effect of the compound on the inhibition of fat accumulation in liver tissue was evaluated.

After necropsy, a certain amount of liver tissue was taken and crushed, followed by Lysis buffer containing 10% Protease inhibitor cocktail (Sigma Co.) (25 mM Tris / pH 7.4, 50 mM NaCl, 0.5% Na-Deoxycholate, 2% Nonidet P-40, 1 ml of 0.2% SDS) was added and the mixture was allowed to stand at room temperature for 10 minutes, followed by centrifugation at 10,000 rpm for 5 minutes, and the supernatant was taken. The obtained liver tissue extract was quantified for protein using a micro BCA assay kit (Micro BCA assay kit, Thermo Scientific, cat # 23232). Triglycerides in liver tissue were measured using the enzyme coloration method (BioVision # K622-100), and then corrected by the amount of protein.

As a result, the compound according to the present invention was found to significantly reduce the triglyceride level significantly increased due to the high fat diet (Fig. 6). These results suggest that the compound according to the present invention can be used as a dietary supplement for preventing and treating fatty liver or improving symptoms of fatty liver.

6) Blood LDL  Measure

Increased adipose tissue due to high fat diet is closely related to deterioration of blood lipid levels, such as increased blood cholesterol, especially LDL cholesterol, and these measurements are important indicators of adult diseases such as hypertension, diabetes, hyperlipidemia. In order to determine the degree of improvement in blood LDL cholesterol level of the compound according to the present invention, cholesterol in serum was measured. As a result, while the high-fat diet increased the LDL cholesterol level in the blood, it was confirmed that the LDL cholesterol level was significantly improved in the group administered with the compound according to the present invention (Fig. 7). These results suggest that the compound according to the present invention can be used as a dietary supplement for treating hyperlipidemia or improving symptoms of hyperlipidemia by lowering blood lipids.

< Example  3> High sugar diet  Hypoglycemic effect in the given mice

Among the compounds of the present invention showing a high blood sugar lowering effect in a high-fat diet model, the mouse was provided with a high sucrose diet for 6 weeks in order to further confirm the hypoglycemic effect of the high sugar diets of the compounds of Formula 1 and Formula 3 Blood glucose was measured after 8 weeks of oral administration of Compound 1 and Formula 3 for 8 weeks. Mice provided with a high glycemic diet significantly increased blood glucose levels compared to the normal group, and after 8 weeks of administration of the compounds of Formulas 1 and 3 according to the present invention, the mice were restored to almost normal levels (FIG. 8). Through this, it was confirmed that the compound of Formula 1 and Formula 3 has a hypoglycemic effect in mice with elevated blood sugar due to high glycemic diet. From these results, the compound according to the present invention can be used as a dietary supplement for treating diabetes or diabetic symptoms.

< Example  4> Acute toxicity test

In order to test the acute toxicity of the compound according to the present invention was carried out based on the single dose toxicity test method of Table 1 of the Toxicological Standards of Food and Drug Safety Notice. Compounds 1 to 3 were administered once at 2,000 mg / kg dose and observed for 3 days in 6-week-old male and female SD rats, and 3 days after compound 1 was administered to male rats at 1,000 mg / kg dose based on mortality. Mortality, clinical symptoms, body weight and necropsy findings were observed as indicators of toxicity during the study. As a result, two of three male rats to which the compound of formula 1 according to the present invention was administered at a dose of 2,000 mg / kg died and no death was observed in other animals. As a result, LD ₅₀ of the compound of Formula 1 was found to be> 2,000 mg / kg in females and 1,000-2,000 mg / kg in males. The LD ₅₀ of 3 compounds was determined to be> 2,000 mg / kg in all males. Therefore, the compound according to the present invention did not show a toxicity change upon oral administration up to a dose of 1,000 mg / kg in rats, it can be seen that it is a safe drug and useful as a dietary supplement.

< Formulation example  1> Preparation of tablets

2-allyl-p-phenylphenol 60 mg

Calcium Silicate 20 mg

Lactose 130 mg

100 mg of microcrystalline cellulose

Magnesium stearate appropriate amount

Talc proper amount

Coating amount

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

< Formulation example  2> Preparation of capsule

6-allyl-o, o′-biphenyl 60 mg

Lactose 160 mg

Starch

The above components are mixed in accordance with a conventional method for producing a capsule, and filled in a gelatin capsule to prepare a capsule.

< Formulation example  3> Preparation of soft capsule

2,2'-diallyl-p, p'-biphenol 60 mg

Tween 80 175mg

PEG400 190mg

Propylene carbonate 100mg

Propylene gylcol appropriate amount

Poloxamer 407 Correct

According to a conventional soft capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

< Formulation example  4> Manufacturing of beverages

0.26% by weight of honey, 0.0002% by weight of thioctoamide, 0.0004% by weight of nicotinic acid, 0.0001% by weight of riboflavin sodium hydrochloride, 0.0001% by weight of pyridoxine hydrochloride, 0.001% by weight of inositol, 0.002% by weight of orthoic acid and 89.7362% by weight of water, 2, The beverage was prepared by the conventional method which mix | blended 10 weight% of 2'- diallyl-p, p'- biphenol.

Claims (2)

A pharmaceutical composition for treating and preventing obesity, fatty liver or hyperlipidemia, comprising a hydroxybiphenyl derivative represented by the following Chemical Formula 1 or 2 and a pharmaceutically acceptable carrier.
&Lt; Formula 1 >

(2)

Health functional food for the improvement of the symptoms of obesity, fatty liver or hyperlipidemia comprising a compound of formula 1 or 2 which is a hydroxybiphenyl derivative.
&Lt; Formula 1 >

(2)

Images