KR20090016804A - Novel benzyl ester compounds and the composition comprising the same for preventing and treating of obesity, diabetes and hyperlipidemia - Google Patents

Abstract

A benzyl ester compound derivative is provided to reduce the body fat and the blood lipid by suppressing the lipid synthesis and cholesterogenesis and increasing the beta oxidation of a fatty acid, and to ensure excellent effects of preventing and treating obesity, diabetes and hyperlipidemia. A benzyl ester compound or a pharmaceutically allowable salt has a structure of the following chemical formula (I). In the chemical formula (I), R1, R2, R3 and R4, are independently at least one substitution radical selected from the group consisting of hydrogen, -OH, amine or methoxy group; the formula (a) is a single bond or a double bond; and n is an integer of 0-10.

Description

Novel benzyl ester compounds and the composition comprising the same for preventing and treating of obesity, diabetes and hyperlipidemia

The present invention relates to a novel benzyl ester compound and a composition for preventing and treating obesity, diabetes and hyperlipidemia containing the same as an active ingredient.

Document 1 Hardie et al, FEBS Lett . 546 , pp 113-120, 2003

[2] David Carling et al, TRENDS in Biochemical Sciences 29 , pp 18-24, 2004

Juleen R. et al, J Appl Physiol . 93 , pp 773-781, 2002

[4] Carmenrodrgue et al, NEJM . 341 , pp 1097-1105, 1999

[Reference 5] Ha J. et al, Mol Pharmacol , 72 , pp 62-72, 2007

6, Holman, RR et al, Metabolism , 55, S2-5,2006; Pickup, JC et al., Blackwell Scientific Publ . London, pp 462-476, 1991

Reference 7 Innerfield, RJ, New Engl . J. Med . , 334 , pp 1611-1613, 1996

Schlattner U, et al, J Biol. Chem . 279 , pp 43940-51, 2004

9 Somwar R et al, Clin Ther . 20 , pp 125-40, 1998

Document 10 Halseth AE et al, Biochem Biophys Res Commun . 294 , pp 798-850, 2002

      The present invention relates to a novel benzyl ester compound and a composition for preventing and treating obesity, diabetes and hyperlipidemia containing the same as an active ingredient.

AMPK (AMP-Activated Protein Kinase) is a kinase that plays a very important role in maintaining the energy balance in the body by controlling the carbohydrate metabolism and lipid synthesis metabolism in the cell by sensing the energy state in the cell. AMPK is an alpha subunit of AMPK caused by AMPK kinase when the ATP content of the cell's bioenergy is sharply lowered, that is, when the ATP: AMP ratio in the cell decreases significantly after heavy exercise or ingestion of food for a long time. The threonine residue 172 of the subunit is phosphorylated and converted into an enzyme in an activated form. As a result, beta oxidation (β-oxidation) metabolism, which is a process of glycolysis and body fat decomposition, is actively performed to generate ATP, a bioenergy (Hardie et al, FEBS). Lett . 546 , pp 113-120, 2003), on the contrary, inhibiting ATP-consuming fat synthesis or cholesterol synthesis (David Carling et al, TRENDS). in Biochemical Sciences 29 , pp 18-24, 2004). In addition, when AMPK is activated, it promotes the absorption of glucose into the cell, thereby increasing the production of substances and ATP necessary for maintaining the body, thereby promoting the lowering of blood glucose.

As a specific metabolic change according to the AMPK activation as described above, phosphorylated activated form of AMPK is a core enzyme of fatty acid synthesis, ACC (acetyl-CoA carboxylase) producing malonyl-CoA and HMG-CoA which is a core enzyme of cholesterol synthesis. By directly phosphorylating reductase proteins, these enzyme activities are lowered, thereby inhibiting fat and cholesterol synthesis. In addition, when the concentration of malonyl-CoA decreases due to a decrease in ACC activity due to increased phosphorylation of ACC, fatty acid inflow is rapidly increased into the mitochondria where beta oxidation, a lipolysis process, is promoted, thereby promoting fat decomposition. do. On the other hand, AMPK activation leads to a decrease in blood glucose because the movement of glucose uptake pathways such as GLUT4 (glucose transporter 4) increases to the cell membrane irrespective of insulin action (Juleen R. et al, J Appl). Physiol . 93 , pp 773-781, 2002).

The obesity population is increasing rapidly around the world, creating many health problems. Obese people have a high risk of diabetes, hypertension, hyperlipidemia, angina, stroke, metabolic syndrome, sleep apnea, osteoarthritis, back pain, stroke, breast cancer, colon cancer, and other diseases are known to be high. Obesity is also a direct cause of mortality and has been reported in several epidemiologic studies (Carmenrodrgue et al, NEJM . 341 , pp 1097-1105, 1999).

Currently, there are two types of anti-obesity drugs on the market, which are fat absorption inhibitors and appetite suppressants. In 2001, the market size increased to about US $ 1.3 billion. However, anti-obesity drugs based on metabolic control functions such as AMPK activators are not yet available. It is not being developed (Ha J. et al, Mol Pharmacol , 72 , pp 62-72, 2007).

Diabetes mellitus is a disease of high insulin and metabolic abnormalities persist due to lack of insulin action, and is highly likely to cause vascular complications in the future. Diabetes is largely divided into insulin-dependent type 1 diabetes and insulin-independent type 2 diabetes.

Diabetes treatment agents currently used in clinical practice are largely known as recombinant insulin, sulfonylurea, metformin, thiazolidinedion, and alpha-glucosidase inhibitor. . However, sulfonyl urea drugs secrete insulin even when insulin is not needed in vivo and risk of hypoglycemia (Holman, RR et al, Metabolism , 55, S2-5,2006; Pickup, JC et al., Blackwell Scientific Publ . London, pp 462-476, 1991), biguanide metformin inhibits hepatic biosynthesis and increases insulin receptors, but causes gastrointestinal disorders and is toxic to death after prolonged administration (Innerfield, RJ, New Engl . J. Med . , 334 , pp 1611-1613, 1996). Thiazolidinedione drugs include rosiglitazome (Avandia) and pioglitazone (Actose Actose), which help insulin function in muscle and adipose tissue and have an excellent effect on blood sugar control. However, side effects can cause hepatotoxicity, so be sure to take regular liver function tests while taking medication. Troglitazone, the first of its class, has been discontinued due to hepatotoxicity, and this class of drugs can also cause obesity. Recently released alpha-glucosidase inhibitors primarily delay the digestion and absorption of complex carbohydrates in the small intestine and reduce postprandial hyperglycemia and hyperinsulinemia, a hallmark of insulin-independent diabetes. However, abdominal bloating, nausea, and diarrhea are typical side effects. Recently, liver damage has been reported. Therefore, there is an urgent need for the development of drugs that minimize the side effects mentioned.

In general, obesity is accompanied by hyperlipidemia and the probability of developing diabetes is very high. This is because type 2 diabetes appears by causing insulin resistance to abnormally increased body lipid content due to obesity. Therefore, if there is an excellent anti-obesity effect that promotes body fat breakdown, and at the same time, a substance that can reduce blood glucose concentration independently of insulin, it can be used as a therapeutic agent for metabolic diseases such as obesity, hyperlipidemia and diabetes treatment . Accordingly, the present inventors have found that MHMB (8-methylnonanoic acid 3-hydroxy-4-methoxy benzyl ester) and its derivatives activate the AMPK enzyme, thereby inhibiting fat synthesis and cholesterol synthesis, and at the same time increasing beta oxidation of fatty acids. Not only does it accelerate the degradation, which leads to a decrease in blood lipids and body fat, but also improves insulin resistance due to a decrease in body fat, and increases cell membrane migration such as GLUT4 by activation of AMPK in an insulin-independent manner. The present invention has been completed by confirming that the composition of the present invention can be usefully used as a component of food, medicine and feed having excellent efficacy in the prevention, inhibition and treatment of obesity, hyperlipidemia and diabetes.

In order to achieve the above object, the present invention provides a novel benzyl ester compound derivative or a pharmaceutically acceptable salt thereof having a novel structure represented by the following general formula (I):

                                       (I)

In the above formula,

R ₁ , R ₂ , R ₃ and R ₄ are each independently one or more substituents selected from the group consisting of hydrogen, hydroxy groups, amine groups or methoxy groups.

) 은 단일결합 또는 이중결합일 수 있으며,(

) May be a single bond or a double bond,

n is an integer of 0-10.

Preferred compound groups among the above general formula (I) compounds are R ₁ , R ₂ , R ₃ and R _{4, which} are hydrogen atoms, hydroxy groups, or amine groups, and n is an integer of 1 to 5, and is more preferable. In the compound group, R ₁ , R ₂ , R _3, and R ₄ are hydrogen atoms or hydroxy groups, n is an integer of 3 or 4, and the most preferred compound is MHMB (8-methylnonanoic acid 3-hydroxy- 4-methoxy benzyl ester) compound.

       In another aspect, the present invention provides an AMP-activated protein kinase (hereinafter, referred to as "AMPK") enzyme activator using the benzyl ester compounds as an active ingredient.

       The compounds of the present invention represented by the general formula (I) may be prepared as pharmaceutically acceptable salts according to methods conventional in the art.

       As salts are acid addition salts formed with pharmaceutically acceptable free acids. Acid addition salts are prepared by conventional methods, for example by dissolving a compound in an excess of aqueous acid solution and precipitating the salt using a water miscible organic solvent, such as methanol, ethanol, acetone or acetonitrile. Equal molar amounts of the compound and acid or alcohol (eg, glycol monomethylether) in water can be heated and the mixture can then be evaporated to dryness or the precipitated salts can be suction filtered.

In this case, organic acids and inorganic acids may be used as the free acid, hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, etc. may be used as the inorganic acid, and methanesulfonic acid, p -toluenesulfonic acid, acetic acid, trifluoroacetic acid, Citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid ( gluconic acid), galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanic acid, hydroiodic acid, and the like.

Bases can also be used to make pharmaceutically acceptable metal salts. An alkali metal or alkaline earth metal salt is obtained by, for example, dissolving a compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and then evaporating and drying the filtrate. At this time, as the metal salt, it is particularly suitable to prepare sodium, potassium or calcium salts, and the corresponding silver salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (for example, silver nitrate).

Pharmaceutically acceptable salts of the above compounds of formula (I) include salts of acidic or basic groups which may be present in compounds of formula (I), unless otherwise indicated. For example, pharmaceutically acceptable salts include sodium, calcium and potassium salts of the hydroxy group, and other pharmaceutically acceptable salts of the amino group include hydrobromide, sulfate, hydrogen sulphate, phosphate, hydrogen phosphate, dihydrogen Phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate) and p -toluenesulfonate (tosylate) salts, which are known in the art. It can be prepared through.

      Another object of the present invention is to provide a method for preparing the compound of Formula (I), which may be chemically synthesized by the method shown in the following schemes, but is not limited thereto.

 The following reaction schemes represent the preparation method of the representative compounds of the present invention in stages of manufacture. Various compounds of the present invention may be prepared by small changes such as changing the reagents, solvents, and reaction sequences used in the synthesis of Scheme (1). have. Some compounds of the present invention were synthesized according to procedures not included in the scope of Scheme 1, and detailed synthesis procedures for these compounds are described in their respective examples.

      Scheme (1) shows a two-step process for preparing a commercial compound or a compound synthesized by known methods.

      In the first step, the activator is reacted in a solvent such as methyl nonanoic acid (1) with anhydrous methylene chloride such as thionyl chloride to obtain an activated halide (2). The benzyl alcohol derivative of general formula (II) may be reacted under an inert solvent such as a pyridine solution to prepare a novel benzyl ester compound derivative of general formula (I), but is not limited thereto.

     The composition containing the general formula (I) compounds obtained by the above method as an active ingredient inhibits fat synthesis and cholesterol synthesis by activating the AMPK enzyme, and at the same time increases the beta oxidation of fatty acids to promote lipolysis, thereby improving blood lipids and body fat. In addition to improving insulin resistance due to the reduction of body fat, and the increase of cell membrane movements such as GLUT4 by AMPK activation by insulin-independent method, it has an excellent effect on lowering blood sugar, thus preventing and suppressing obesity, hyperlipidemia and diabetes. And food, pharmaceutical and feed additives having excellent efficacy in treatment.

      Accordingly, the present invention provides a pharmaceutical composition for the prevention and treatment of obesity, hyperlipidemia and diabetes containing the novel benzyl ester compound derivative represented by the general formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient. .

      More specifically, for the prevention and treatment of obesity, hyperlipidemia and diabetes through the activation of AMPK enzyme containing a novel benzyl ester compound derivative represented by the general formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient Provide a pharmaceutical composition.

       The present invention also provides an AMPK enzyme activator containing a novel benzyl ester compound derivative represented by the general formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient.

     Compositions comprising a compound of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

Compositions comprising the compounds of the invention are each formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories, and sterile injectable solutions, according to conventional methods. Carriers, excipients and diluents that may be included in the composition comprising the extract include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, Calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations may contain at least one excipient such as starch, calcium carbonate, sucrose, or the like. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Oral liquid preparations include suspending agents, liquid solutions, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Preferred dosages of the compounds of the present invention depend on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, but may be appropriately selected by those skilled in the art. However, for the desired effect, the compound of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably at 0.001 to 10 mg / kg. Administration may be administered once a day or may be divided several times. Therefore, the above dosage does not limit the scope of the present invention in any aspect.

The compounds of the present invention can be administered to mammals such as mice, mice, livestock, humans, and the like by various routes. All modes of administration can be expected, for example by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection. Pharmaceutical dosage forms of the compounds of the present invention may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable collection.

In addition, the present invention is to prevent the obesity, hyperlipidemia and diabetes prevention of obesity, hyperlipidemia and diabetes containing a novel benzyl ester compound of formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient showing the effect of preventing and improving And it provides a dietary supplement for improvement.

      Compounds of the present invention can be used in a variety of drugs, food and beverages for the prevention and improvement of obesity, hyperlipidemia and diabetes. Examples of the food to which the compound of the present invention may be added include various foods, beverages, gums, teas, vitamin complexes, health supplements, and the like, and may be used in the form of powders, granules, tablets, capsules, or beverages. have.

Since the compound of the present invention has little toxicity and side effects, it is a drug that can be used safely even for long-term administration for the purpose of prevention.

The compounds of the present invention may be added to food or beverages for the purpose of preventing and improving obesity, hyperlipidemia and diabetes. At this time, the amount of the compound in the food or beverage is generally added to the health food composition of the present invention to 0.01 to 15% by weight of the total food weight, the health beverage composition is 0.02 to 30 g based on 100 ml, preferably Can be added in a ratio of 0.3 to 10 g.

The health beverage composition of the present invention, in addition to containing the compound as an essential ingredient in the indicated proportions, has no particular limitation on the liquid component and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. Examples of the above-mentioned natural carbohydrates are conventional monosaccharides such as disaccharides such as glucose and fructose, such as maltose, sucrose and the like, and polysaccharides such as dextrin, cyclodextrin and the like. Sugars and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the compounds of the present invention, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid And salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. The compositions of the present invention may also contain pulp for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected from the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

      In addition, the present invention is to prevent the obesity, hyperlipidemia and diabetes prevention of obesity, hyperlipidemia and diabetes containing a novel benzyl ester compound of formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient showing the effect of preventing and improving And animal feed additives useful for improvement and feed comprising the same.

      The animal feed additive compound may be 20 to 90% high concentrate or powder or granule form.

Compounds for animal feed additives of the present invention are organic acids such as citric acid, fumaric acid, adipic acid, lactic acid, malic acid, phosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate, polyphosphate (polymeric phosphate), polyphenols, catechins (catechin) ), Alpha-tocopherol, rosemary extract (rosemary extract), vitamin C, green tea extract, licorice extract, chitosan, tannic acid, phytic acid and the like may further include any one or more than one.

Animal feed additives containing a compound of the present invention and feed containing the same as a secondary component, various supplements such as amino acids, inorganic salts, vitamins, antibiotics, antibacterial substances, antioxidants, antifungal enzymes, live microbial preparations, grains, for example Milled or crushed wheat, oats, barley, corn and rice; Vegetable protein feed, such as rape, soybeans and sunflower; Animal protein feeds such as blood meal, meat meal, bone meal and fish meal; Sugars and dairy products, for example, a dry ingredient consisting of various powdered milk and whey powder, and a drying additive, all mixed together with a liquid component, and a component which becomes a liquid after heating, that is, a lipid, for example, an animal liquefied by heating. In addition to the main components such as fats and vegetable fats can be used with substances such as nutritional supplements, digestion and absorption enhancers, growth promoters, disease prevention agents and the like.

The animal feed additive compound may be administered to an animal alone in combination with other feed additives in an edible carrier.

In addition, the compound for animal feed additives can be easily administered as a top dressing or directly mixed with the animal feed or separately from the feed, in a separate oral formulation, by injection or transdermal or in combination with other ingredients. Typically, single or divided daily dosages can be used as is well known in the art.

When the compound for animal feed additive is administered separately from the animal feed, the dosage form of the compound, as is well known in the art, may be prepared in an immediate release or sustained release formulation by combining them with a non-toxic pharmaceutically acceptable edible carrier. have. Such edible carriers may be solid or liquid, for example corn starch, lactose, sucrose, soy flakes, peanut oil, olive oil, sesame oil and propylene glycol. When a solid carrier is used, the dosage form of the compound may be tablets, capsules, powders, torokies or sugar-containing tablets or top dressings in microdisperse form. If a liquid carrier is used, it may be in the form of soft gelatin capsules or syrups or liquid suspensions, emulsions or solutions. In addition, the dosage form may contain auxiliaries such as preservatives, stabilizers, wetting or emulsifying agents, solution promoters and the like.

In addition, the animal feed in which the compound of the present invention is included as an animal feed additive may be any protein-containing organic cereal meal conventionally used to meet the dietary needs of animals. Such protein-containing flours typically consist mainly of corn, soy flour, or corn / bean flour mixtures.

The animal feed additive may be used by adding to the animal feed by dipping, spraying or mixing.

The invention is applicable to a number of animal diets, including mammals, poultry and fish. More specifically, the diet is commercially important mammals such as pigs, cows, sheep, goats, laboratory rodents (rats, mice, hamsters and gerbils), furry animals (eg, mink and fox), and Zoo animals (eg monkeys and tailless monkeys), as well as livestock (eg cats and dogs). Commercially important poultry typically include chickens, turkeys, ducks, geese, pheasants and quails. Commercially raised fish, such as trout, may also be included.

In the animal feed blending method comprising the compound according to the present invention, the compound is incorporated into the animal feed in an amount of about 1 g to 100 g per kg of feed on a dry weight basis.

In addition, the feed mixture is thoroughly mixed and then a viscous granulated or granular material is obtained depending on the degree of grinding of the components. It is fed as a mash or formed into the desired discrete shape for further processing and packaging. At this time, it is preferable to add water to the animal feed and then go through the usual pelletization, expansion, or extrusion process to prevent separation during storage. Excess water may be removed to dryness.

      The novel benzyl ester compound derivatives of the present invention or pharmaceutically acceptable salts thereof inhibit lipogenesis and cholesterol synthesis by activating the AMPK enzyme, and at the same time increase the beta oxidation of fatty acids to promote lipolysis, thereby reducing blood lipids and body fat. In addition to improving insulin resistance due to a decrease in body fat and increasing cell membrane movements such as GLUT4 by AMPK activation in a insulin-independent manner, it has an excellent effect on lowering blood sugar, thus preventing, inhibiting and preventing obesity, hyperlipidemia and diabetes. It can be usefully used as a food, medicine and feed additive having excellent efficacy in treatment.

     Hereinafter, the present invention will be described in detail by Examples and Experimental Examples.

However, the following Examples and Experimental Examples are only illustrative of the present invention, and the content of the present invention is not limited to the following Examples and Experimental Examples.

Reference Example  1. Reagents and Instruments

Analyzers include ¹ H-NMR (400 MHz) and ¹³ C-NMR (400 MHz) spectrometers (JNM-AL 400, JEOL Ltd., Japan), Melting pointers (Yamako, MD-S3, Japan), Mass spectrometer (MS, PE SCIX API 2000 MS / MS, Canada) was used. Various reagents were used by Aldrich Chemical Co., and other solvents were used without purification of the first or higher reagents. Silica gel (Silica gel, Merck, 230-400 mesh) was used for the purification of the synthesized materials.

Example  1. Preparation of Intermediates

The synthesis of 8-methylnonanoic acid 4-hydroxy-3-methoxybenzyl ester and 8-methylnonanoic acid 3-hydroxy-4-methoxybenzyl ester is as follows.

1-1. 8-methylnonanoic acid (8- methylnonanoic acid ) Synthesis

1-1-1. 3- Hydro -4- Of methoxybenzyl ester (Dihydrocapsiate)  synthesis

1-1-1-1. (6- Carboxhesil ) Triphenylphosphorium  Bromide ((6- carboxhexyl )- triphenylphosphorium  bromide)

      A mixture of 6-bromohexanoic acid (25.8 g, 0.13 mol) and triphenylphosphine (34.5 g, 0.13 mol) was heated at 145 ° C. for 4 hours. The glassy reaction mixture obtained by cooling was triturated with dry chloroform and diluted with ether. The resulting precipitate (58 g, 96%) was recrystallized from a given chloroform to give a white powdery phase (52.8 g, 87%) (6-carboxhesyl) triphenylphosphenyl bromide showing the following physical properties.

¹ H-NMR (200 MHz, CDCl ₃ ): 1.66 (m, 6H), 2.37 (m, 2H), 3.62 (m, 2H), 5.97 (bs, 1H), 7.71-7.84 (m, 15H).

1-1-1-2. (Z) -8- methyl -6- nonenoic acid (8- methyl -6- nonenoic acid )

A mixture of (6-carboxhesyl) triphenylphosphorium bromide (25.0 g, 54.6 mmol) and isobutylaldehyde (10.2 ml, 109.3 mmol) in dry DMF (130 ml) was kept at 0 ° C. N ₂ atm for 15 minutes. To a dry DMF (140 ml) KotBu (tetrabutoxide potassiumsulf) 13.2 g, 112.1 mmol) was added to a suspension. Stir vigorously at room temperature for 15 hours. The resulting slurry was poured into ice water (150 ml) and cooled. The precipitated triphenylphosphine oxide was removed by suction filtration. The filtration process was performed by washing with benzene (40ml × 2) and 2M hydrochloric acid. The product was extracted with washed ether (30 ml × 4) with saturated brine (20 ml × 4) and dried over anhydrous Na ₂ SO ₄ . The dried product was distilled off to obtain an oily form (52.8 g, 87%) (Z) -8-methyl-6-nonenoic acid having the following physical properties.

 b.p .: 109-110 ℃

IR (neat): 3000-2500 (COOH), 740 (C = C) cm- ¹ ;

¹ H NMR (200 MHz, CDCl ₃ ): 0.94 (d, J = 8.0 Hz, 6H), 1.27-1.46 (m, 2H), 1.57-1.72 (m, 2H), 2.00-2.11 (m, 2H), 2.35 (t, J = 6.0 Hz, 2H), 2.53-2.62 (m, 1H), 5.11-5.27 (m, 2H);

EI-MS (m / z; relative intensit): 170 (M ⁺ , 14), 152 (M ⁺ -18, 13), 137 (19), 109 (13), 95 (20) 81 (22), 69 100, 55 (77), 41 (85).

1-1-2. 8-methylnonanoic acid

(Z) -8-methylnonanoic acid (3.28 g, 19.3 mmol) dried over 100 ml of ethanol was heated to 20 ° C N ₂ for 20 minutes. It was added to a supernatant of 10% Pd-carbon (1.6 g) and ammonium formate (6.08 g, 96 mmol) dried with 100 ml of ethanol under atmospheric pressure. After stirring vigorously at room temperature for 18 hours, the resulting slurry was removed with a suction filter. The filtrate was distilled under reduced pressure to obtain (Z) -8-methyl-6-nonanoic acid in the form of an oil having the following physical properties (3.01 g, 89%).

¹ H NMR (200 MHz, CDCl ₃ ): 0.85 (d, J = 6.4 Hz, 6H), 1.16-1.65 (m, 11H), 2.22-2.29 (m, 2H), 10.35 (bs, 1H).

1-2. Preparation of 8-methylnonanoic acid 4-hydroxy-3-methoxybenzyl ester

(Z) -methylnonanoic acid (3.4 g, 19.7 mmol) and thionyl chloride (3.54 g, 30 mmol) obtained in the above step were added to dry MC (10 mL of dichloromethane) and dried with CaCl ₂ at room temperature, and then dried at room temperature. Stir overnight. The obtained reaction was dehydrated under reduced pressure to give a brown oil. To this oil was added dropwise 40 mL of 3-hydroxy-4-methoxybenzyl alcohol (8.16 g, 53 mmol) pyridine solution. The mixture was stirred at 0 ° C for 2 h. To the reaction was added water and acidified 2N HCl, then fractionated three times with 100 mL EtOAc each. The combined supernatant EtOAc layer was washed with water, dried over anhydrous sodium sulfate and filtered. In order to remove the residue of the obtained filtrate, dehydration was performed under reduced pressure, and the residue was purified by silica gel column chromatography. 8-methylnonanoic acid 4-hydroxy-3-methoxybenzyl which is extracted with n-hexane / ethyl acetate (70:10 v / v) and shows the following physical properties of colorless oil (1.2 g, 20.1% yield). An ester was obtained.

¹ H NMR (200 MHz, CDCl ₃ ): 0.84 (d, J = 6.4 Hz, 6H), 1.14-1.25 (m, 8H), 1.43-1.62 (m, 3H), 2.32 (t, J = 7.6 Hz, 2H ), 3.90 (s, 3H), 5.02 (s, 2H), 5.64 (s, 1H), 6.88 (m, 3H).

Example  2. 8- Methylnonanoic acid  3- Hydroxy -4- Of methoxybenzyl ester (MHMB)  Produce

2-1. (6- Carboxhesil ) Triphenyl phosphphenyl  Bromide

     A mixture of 6-bromohexanoic acid (25.8 g, 0.13 mol) and triphenylphosphine (34.5 g, 0.13 mol) was heated at 145 ° C. for 4 hours. The cooled glassy mixture was triturated with dry chloroform and diluted with ether. The precipitate obtained (58 g, 96%) was recrystallized from a given chloroform to prepare (6-carboxhesyl) triphenylphosphphenyl bromide in the form of white powder (52.8 g, 87%) having the following physical properties.

¹ H NMR (200 MHz, CDCl ₃ ); 1.66 (m, 6H), 2.37 (m, 2H), 3.62 (m, 2H), 5.97 (bs, 1H), 7.71-7.84 (m, 15H).

2-2. (Z) -8- methyl -6- Of nonenoic acid (a)  synthesis

A mixture of (6-carboxhesyl) triphenylphosphorium bromide (25.0 g, 54.6 mmol) and isobutylaldehyde (10.2 ml, 109.3 mmol) in dry DMF (130 ml) was dried for 15 minutes at 0 ° C. N ₂ atm. 140 ml) was added to a KotBu (13.2 g, 112.1 mmol) suspension. Stir vigorously at room temperature for 15 hours. The resulting slurry was poured into ice water (150 ml) and cooled. Precipitated triphenylphosphine oxide was removed by suction filtration. The filtration process was washed with benzene (40ml × 2) and 2M hydrochloric acid. The resulting product was extracted with ether (30 ml × 4) washed with saturated brine (20 ml × 4) and then dried over anhydrous Na ₂ SO ₄ . The dried product was distilled off to obtain (Z) -8-methyl-6-nonenoic acid of (52.8 g, 87%) in the form of an oil having the following physical properties.

b.p. 109-110 ° C .;

IR (neat): 3000-2500 (COOH), 740 (C = C) cm- ¹ ;

¹ H NMR (200 MHz, CDCl ₃ ): 0.94 (d, J = 8.0 Hz, 6H), 1.27-1.46 (m, 2H), 1.57-1.72 (m, 2H), 2.00-2.11 (m, 2H), 2.35 (t, J = 6.0 Hz, 2H), 2.53-2.62 (m, 1H), 5.11-5.27 (m, 2H);

EI-MS (m / z; relative intensity): 170 (M ', 14), 152 (M'-18, 13), 137 (19), 109 (13), 95 (20), 81 (22), 69 (100), 55 (77), 41 (85).

2-3. (Z) -8- Methylnonanoic acid (b)

(Z) -8-methylnonanoic acid (3.28 g, 19.3 mmol) dried over 100 ml of ethanol was heated to 20 ° C N ₂ for 20 minutes. Under atmospheric pressure, a supernatant of 10% Pd-carbon (1.6 g) and ammonium formate (6.08 g, 96 mmol) dried with 100 ml of ethanol was added. After stirring vigorously at room temperature for 18 hours, the resulting slurry was removed with a suction filter. The filtration process was carried out by distillation under reduced pressure to leave a residue of filtration (3.01 g, 89%) to obtain (52.8 g, 87%) (Z) -8-methyl-6-nonanoic acid having the following physical properties.

¹ H NMR (200 MHz, CDCl ₃ ): 0.85 (d, J = 6.4 Hz, 6H), 1.16-1.65 (m, 11H), 2.22-2.29 (m, 2H), 10.35 (bs, 1H).

2-4. 8- Methylnonanoic acid  3- Hydroxy -4- Methoxybenzyl ester

(Z) -Methylnonanoic acid (3.4 g, 19.7 mmol) and thionyl chloride (3.54 g, 30 mmol) were added to a dry MC (10 mL) and dried with CaCl ₂ at room temperature, and stirred overnight at room temperature. After dehydration under reduced pressure a brown oil was obtained. To the obtained oil was added dropwise 40 mL of 3-hydroxy-4-methoxybenzyl alcohol (8.16 g, 53 mmol) pyridine solution. The reaction mixture was stirred at 0 ° C. for 2 hours. To the reaction was added water and acidified 2N HCl, then fractionated three times with 100 mL EtOAc each. The combined supernatant EtOAc layer was washed with water, dried over anhydrous sodium sulfate and filtered. In order to remove the residue of the obtained filtrate, dehydration was performed under reduced pressure, and the residue was purified by silica gel column chromatography. 8-methylnonanoic acid 4-hydroxy-3-methoxybenzyl which is extracted with n-hexane / ethyl acetate (70:10 v / v) and shows the following physical properties of colorless oil (1.2 g, 20.1% yield). An ester was obtained.

¹ H-NMR (200 MHz, CDCl ₃ ): 0.84 (d, J = 6.4 Hz, 6H), 1.14-1.25 (m, 8H), 1.43-1.62 (m, 3H), 2.32 (t, J = 7.6 Hz, 2H), 3.90 (s, 3H), 5.02 (s, 2H), 5.64 (s, 1H), 6.88 (m, 3H).

   The spectral data of the compounds obtained by the analytical means are shown in Table 1, respectively.

Experimental Example  One. AMPK Activation efficacy

In order to test the AMPK enzyme activation of the compounds obtained in the above examples, the methods described in the literature were applied to determine the efficacy of the compounds as follows (Schlattner U, et al, J Biol). Chem . 279 , pp 43940-51, 2004).

   Each of the 30 μg / ml MHMB and Dihydrocapsiate was treated with L6 myotube cells (American Type Culture Collection, Manassas VA.) As shown in FIG. 2 to activate AMPK antibodies (Cell Signaling Technology, Beverly, MA). In order to confirm the increase in activation of the AMP-activated protein kinase (hereinafter referred to as "AMPK") enzyme with treatment time, the phosphorylation of threonine residue 172 in the alpha (a) -subunit of AMPK was increased. (Figure 2) Shown by western blot assay.

     As a result of the experiment, as shown in Figure 2, both materials showed an increase in time, but it was confirmed that the time-effectiveness in MHMB. As a result, MHMB and Dihydrocapsiate were found to be beneficial for AMPK activation.

Experimental Example 2 Measurement of glucose uptake of L6 myotube cells

To determine the glucose uptake in L6 myotube cells of the compounds obtained in the above examples, the methods described in the literature were applied to determine the efficacy of the compounds as follows (Somwar R et al, Clin Ther. 20 , 125 -40, 1998).

Incubated fully differentiated L6 myotube cells in serum-free medium (GIBCO, Auckand NZ) for 6 hours, then treated with 10 µg / ml MHMB and 30 µg / ml Dihydrocapsiate for 2 hours, respectively, and then 10 mM unlabeled 2 The reaction was carried out in HEPES-saline buffer containing -deoxyglucose and 10 mM 2-deoxy- [ ³ H] -glucose (1 mCi / ml) for 20 minutes. After washing the cells three times with ice-cold PBS, the cells were lysed using 1N NaOH and cpm was measured with a scintillation counter (Packard Co. 1600PR).

In this case, intracellular non-specific glucose uptake was measured by HEPES-saline buffer containing 10 mM cytochalasin B (Sigma Chemical Co. St Louis MO) and removed from the total value.

    As a result, as shown in Figure 3, it was also confirmed that the glucose uptake (glucose uptake) of MHMB was higher than Dihydrocapsiate at the same concentration.

Experimental Example 3 Blood Glucose Measurement in db / db Mice

In order to measure the blood glucose level in the db / db mice of the compounds obtained in the above examples, the efficacy of the compounds was measured as follows (Halseth AE et al, Biochem Biophys Res Commun. 294 , 798-850, 2002).

     Diabetic mice, 5 week old db / db mice (Charles River Laboratories, Inc Germany), divided into 8 mice each at 20 mg / kg (saline) 8-methylnonanoic acid 3-hydro-4-methoxybenzyl ester And 8-methylnonanoic acid 3-hydroxy-4-methoxybenzyl ester-administered group and saline-administered group of the same volume (control) were administered to the mice for 4 weeks (20 mg / kg of body weight once a day) Changes in body weight, appetite, and blood sugar were examined.

    As shown in FIG. 4, MHMB and Dihydrocapsiate-administered groups decreased 37.87% and 40.22% (198.00 ± 22.37, 190.50 ± 15.90, 318.67 ± 45.84) at 3 weeks, respectively, compared to the saline group. 41.47% and 38.19% (265.13 ± 28.82, 280.00 ± 36.15, 453.00 ± 41.26) were decreased.

Experimental Example 4 Measurement of Total Cholesterol and Triglyceride Levels in ob / ob Mice

In order to determine the total cholesterol and triglyceride concentration in the ob / ob mice of the compounds obtained in the above example, the efficacy of the compounds was measured as follows (Halseth AE et al, Biochem Biophys Res Commun. 294 , 798-850, 2002).

    Five obese ob / ob mice (The Jackson Laboratory, Bar Harbor, ME, Inc Germany) were divided into 8 mice each with 20 mg / kg (saline) of MHMB and Dihydrocapsiate and the same volume of saline (control). Was administered for 8 weeks to investigate the change in weight, appetite, total cholesterol, and triglyceride of the mice.

     As shown in FIG. 5, MHMB and Dihydrocapsiate-treated groups showed 10.06% and 7.38% (40.32 ± 1.02, 41.52 ± 0.77, 44.83 ± 1.17) at 5 weeks, and 10.19% and 8.95 at 6 weeks, respectively, in the change of body weight. % (41.97 ± 1.17, 42.55 ± 1.42, 46.73 ± 1.06), 9.70% and 7.96% at 7 weeks (44.13 ± 1.77, 44.98 ± 1.69, 48.87 ± 1.13), 9.25% and 8.26% at 45 weeks (45.92 ± 1.45, respectively) 46.42 ± 1.35, 50.60 ± 1.23).

    Total cholesterol levels decreased by 26.22% and 21.77% (157.25 ± 13.23, 166.73 ± 17.94, 213.13 ± 7.04) in MHMB and dihydrocapsiate groups, respectively, and triglyceride levels were 22.10% and 18.22%, respectively. % (52.34 ± 4.03, 54.95 ± 7.26, 67.19 ± 12.24) was decreased.

Experimental Example 5. Acute Toxicity Test

      In order to test the acute toxicity of the compound of the present invention, according to the regulations of the Food and Drug Administration (KFDA) was tested using male ICR mice (4 weeks old, the central laboratory animals).

      Ten mice in each group were orally administered with a compound of the present invention at a dose of 0.625-2 g / kg and observed for two weeks.

      After administration of the test substance, mortality, clinical symptoms, and changes in body weight were observed, blood biochemical tests were performed, and autopsy was performed to observe abnormalities of organs and thoracic organs.

     As a result, there were no clinical symptoms or dead animals in all animals treated with the test substance, and no toxicity change was observed in weight change, blood test, blood biochemical test and autopsy findings.

The compound of the present invention did not show a toxic change when administered orally to a dose of 2g / kg in mice, 1-1000 mg / kg orally administered to humans, 0.2-100 mg / cm ₂ when applied to the skin effect Was determined to be a substance.

Examples of the formulation of the composition are described below, but are not intended to limit the present invention but to explain in detail only.

Formulation example  One. Powder  Produce

MHMB 20 mg

Lactose 100 mg

Talc 10 mg

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

Formulation example  2. Preparation of Tablets

MHMB 10 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

     After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Formulation example  3. Manufacture of capsule

Dihydrocapsiate 10 mg

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

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

Formulation example  4. Preparation of Injectables

MHMB 10 mg

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

Na ₂ HPO ₄ 12H ₂ O 26 mg

     According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation example  5. Liquid  Produce

Dihydrocapsiate 20 mg

10 g of isomerized sugar

5 g of mannitol

Purified water

      According to the conventional method for preparing a liquid, each component is added and dissolved in purified water, lemon flavor is added, the above components are mixed, purified water is added, the whole is adjusted to 100 ml by adding purified water, and then filled in a brown bottle. The solution is prepared by sterilization.

Formulation example  6. Manufacture of healthy food

MHMB 1000 mg

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

0.2 μg of vitamin B12

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

Folate 50 ㎍

Calcium Pantothenate 0.5mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

     Although the composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. The granules may be prepared and used for preparing a health food composition according to a conventional method.

Formulation example  7. Manufacture of health drinks

Dihydrocapsiate 1000 mg

Citric acid 1000 mg

100 g oligosaccharides

Plum concentrate 2 g

1 g of taurine

Add 900 ml of purified water

     After mixing the above components according to a conventional healthy beverage manufacturing method, and then stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed and sterilized and then refrigerated and stored in the present invention For the preparation of healthy beverage compositions.

     Although the composition ratio is a composition suitable for a preferred beverage in a preferred embodiment, the composition ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, use purpose.

    1 shows 8-methylnonanoic acid 3-hydroxy-4-methoxybenzyl ester (MHMB) structure (A) and 8-methylnonanoic acid 4-hydroxy-3-methoxybenzyl ester (Dihydrocapsiate) A diagram showing a structure (B),

FIG. 2 shows residues of threonine 172 present in the alpha (a) -subunit of AMPK to confirm increased activation of AMPK enzymes with treatment time of L6 muscle cells derived from mice. Phosphorylation is confirmed by antibody staining method,

Figure 3 is a diagram showing the increase in glucose uptake (glucose uptake) in the cells when the sample was treated for 2 hours to L6 muscle cells,

Figure 4 is a diagram showing the changes in dietary intake, body weight and blood sugar when oral administration of the sample to the db / db mice of the diabetic mouse model,

Figure 5 is a diagram showing the changes in feed intake, body weight, total cholesterol and triglycerides when oral administration of the sample in ob / ob mouse model of obese mice.

Claims (9)

A novel benzyl ester compound of the novel structure represented by the following general formula (I) or a pharmaceutically acceptable salt thereof:

                                         (I) In the above formula, R ₁ , R ₂ , R ₃ and R ₄ are each independently one or more substituents selected from the group consisting of hydrogen, hydroxy groups, amine groups or methoxy groups. (

) May be a single bond or a double bond, n is an integer of 0-10. The compound of claim 1, wherein the compound of general formula (I) is R ₁ , R ₂ , R ₃ and R ₄ are hydrogen atoms, hydroxy groups, or amine groups, and n is an integer of 1 to 5, or a pharmaceutically acceptable compound thereof. Possible salts. The compound according to claim 2, wherein the compound of general formula (I) is a compound in which R ₁ , R ₂ , R ₃ and R ₄ are hydrogen atoms or hydroxy groups, and n is an integer of 3 or 4, or a pharmaceutically acceptable salt thereof. . The compound of claim 3, wherein the compound of general formula (I) is MHMB (8-methylnonanoic acid 3-hydroxy-4-methoxy benzyl ester) compound or a pharmaceutically acceptable salt thereof. An AMP-activated protein kinase (AMPK) enzyme activator comprising benzyl ester compounds represented by the general formula (I) of claim 1 as an active ingredient.  The prophylaxis and treatment of obesity, hyperlipidemia and diabetes comprising a novel benzyl ester compound derivative or a pharmaceutically acceptable salt thereof as an active ingredient comprising the benzyl ester compounds represented by the general formula (I) of claim 1 as an active ingredient. Pharmaceutical composition for.  Prevention and improvement of obesity, hyperlipidemia and diabetes containing a novel benzyl ester compound of formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient, which shows the prevention and improvement effect of obesity, hyperlipidemia and diabetes Dietary supplements.  Animals useful for the prevention and improvement of obesity, hyperlipidemia and diabetes, containing as an active ingredient a novel benzyl ester compound derivative of formula (I) or a pharmaceutically acceptable salt thereof, which has an effect of preventing and improving obesity, hyperlipidemia and diabetes Feed additives.  Feed comprising the animal feed additive of claim 8.

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