KR101627546B1 - Protocatechuic acid derivatives with lipase inhibition activity and use therof - Google Patents

Abstract

The present invention relates to a protocatechuic acid derivative with lipase inhibitory activity; and to a use thereof. According to the present invention, a composition with lipase inhibitory activity can be used for common beverages, health supplementary food, or medicines, and thus can have a great effect on the prevention, alleviation, or treatment of obesity. A health food composition for the prevention or alleviation of obesity contains a protocatechuic acid derivative or salt thereof as an active ingredient.

Description

Protocatechuic acid derivatives having lipase inhibitory activity and uses thereof < RTI ID = 0.0 >

The present invention relates to a protocatechuic acid derivative having lipase inhibitory activity and use thereof.

Obesity is usually a lot of weight, but obesity, even if you have a lot of muscle weight can go out a lot of fat tissue in the body is defined as a state of excess of obesity. Obesity, which is caused by unbalance of energy consumption and intake, is applied with exercise therapy, low calorie diet and other medication depending on the degree. Of these, pharmacotherapy reduces food intake, changes nutrition absorption and metabolism, In the direction of increase.

Lipid taken as food is decomposed into free fatty acid and monoacylglycerol by lipase which is secreted from the stomach or pancreas and bile secreted from gallbladder to form bile and micelle structure, And is absorbed into the small intestine mucosal cells and used as an energy source and a constituent of the human body, and the surplus energy is accumulated in the fat cells in the body. One of the most important methods for inhibiting or treating obesity is to inhibit lipase, an enzyme that decomposes lipids, to inhibit lipid digestion and absorption, thereby reducing energy intake through the stomach.

However, in recent years, the development of appetite suppressants, heat stimulants, and fat absorption inhibitors has been progressing, and the usefulness of these substances as a therapeutic agent for obesity has attracted attention, Among them, the lipase, a pancreatic lipase inhibitor, is of interest.

Pancreatic lipase acts as a major enzyme that breaks down triglycerides into 2-monoacylglycerol and fatty acids. A representative known pancreatic lipase inhibitor is Streptomyces tricycline It is known that tetrahydrolipstatin (orlistat), a derivative of lipstatin derived from toxitricini, is currently marketed as an anti-obesity drug and has the best efficacy to inhibit about 30% of the fat ingested . However, despite this efficacy, orlistat is known to have side effects such as gastrointestinal disorders, hypersensitivity, cholestatic disturbances and lipid-soluble vitamin absorption inhibition.

Korean Patent Publication No. 2014-0112747 discloses a pharmaceutical preparation in which the dissolution rate of a lipase inhibitor is improved and a side effect is reduced and a preparation method thereof. Korean Patent No. 1350824 discloses a lipase-specific lipid-peptide substrate library And a lipase inhibitor using the same as an active ingredient. However, as in the present invention, it is known that a protocatechuic acid derivative having lipase inhibitory activity and a use thereof are useful for the treatment and prevention of obesity and lipid metabolism abnormality- It is not known.

The present invention has been made in view of the above-mentioned needs, and it is an object of the present invention to isolate and purify protocatechuic acid as an anti-obesity substance from juice of pine needle and to screen 18 kinds of derivatives having excellent anti- Thereby completing the invention.

In order to accomplish the above object, the present invention provides a process for preparing 3-amino-4-hydroxybenzoic acid; 2,3,4-trihydroxybenzoic acid; 4-nitrobenzoic acid; 4-methoxybenzoic acid; 4-fluorobenzoic acid (4-fluorobenzoic acid); 3,4-dimethoxybenzoic acid; 2,3-dihydroxybenzoic acid; 2,4-dihydroxybenzoic acid; 2,5-dihydroxybenzoic acid (2,5-dihydroxybenzoic acid); 4-hydroxybenzenesulfonic acid; Terephthalic acid; 3,4-dihydroxyphenylacetic acid; 3,4-dihydroxyphenylacetic acid; D - (-) - tartaric acid (D - (-) - tartaric acid); Dihydroxyfumaric acid hydrate; 2-pyrazinesulfonic acid; 4,4'-sulfonyldibenzoic acid; 5-hydroxynicotinic acid; And 4-hydroxy-5-methyl-4-cyclopentene-1,3-dione monohydrate. The present invention provides a health food composition for preventing or ameliorating obesity containing a cathechic acid derivative or a salt thereof as an active ingredient.

The present invention also provides a pharmaceutical composition for preventing or treating obesity, which comprises the protocatechuic acid derivative or a salt thereof as an active ingredient.

The composition containing the protocatechuic acid derivative according to the present invention as an active ingredient can be applied to ordinary beverages, health supplements or medicines, and thus can have a great effect on preventing, improving or treating obesity.

Figure 1 shows the results of analysis of lipase inhibitory activity of protocatechuic acid and its derivatives (compounds 1 to 10).
2 shows the results of analysis of the lipase inhibitory activity of the protocatechuic acid derivatives (Compound 11 to Compound 20).
3 shows the results of analysis of the lipase inhibitory activity of the protocatechuic acid derivatives (compounds 21 to 24).

The present invention relates to 3-amino-4-hydroxybenzoic acid; 2,3,4-trihydroxybenzoic acid; 4-nitrobenzoic acid; 4-methoxybenzoic acid; 4-fluorobenzoic acid (4-fluorobenzoic acid); 3,4-dimethoxybenzoic acid; 2,3-dihydroxybenzoic acid; 2,4-dihydroxybenzoic acid; 2,5-dihydroxybenzoic acid (2,5-dihydroxybenzoic acid); 4-hydroxybenzenesulfonic acid; Terephthalic acid; 3,4-dihydroxyphenylacetic acid; 3,4-dihydroxyphenylacetic acid; D - (-) - tartaric acid (D - (-) - tartaric acid); Dihydroxyfumaric acid hydrate; 2-pyrazinesulfonic acid; 4,4'-sulfonyldibenzoic acid; 5-hydroxynicotinic acid; And 4-hydroxy-5-methyl-4-cyclopentene-1,3-dione monohydrate. The present invention relates to a health food composition for preventing or ameliorating obesity, which comprises a catheter derivative or a salt thereof as an active ingredient.

The composition of the present invention may be added to a health food for the purpose of preventing or improving obesity. In addition, health food means that there is a certain health effect when it is taken, but unlike general medicine, there is an advantage that there is no side effect that can occur when a medicine is taken for a long time by using food as a raw material.

The health food composition is to prevent or ameliorate obesity by inhibiting lipase activity, and the protocatechuic acid derivative preferably includes a carboxyl group or a sulfonic acid group, but is not limited thereto.

In addition, in addition to the above-mentioned active ingredients, there can be further added flavors, natural carbohydrates, nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickening agents, , Glycerin, alcohols, and carbonates used in carbonated beverages.

When the protocatechuic acid derivative of the present invention is used as a food additive, the protocatechuic acid derivative may be added as it is or may be used together with other food or food ingredients, and may be appropriately used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (preventive, healthful or ameliorative treatment). Generally, in the production of foods or beverages, the protocatechuic acid derivatives of the present invention are added in an amount of not more than 15% by weight, preferably not more than 10% by weight based on the raw material. However, in the case of long-term ingestion intended for health and hygiene purposes or for health control purposes, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount in the above- .

There is no particular limitation on the kind of the food. Examples of the food to which the above substances can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include healthy foods in a conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The above-mentioned natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, A carbonating agent used in a carbonated beverage, and the like. In addition, the composition of the present invention may contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination.

The proportion of such additives is not critical, but is generally 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 present invention also relates to a process for the preparation of 3-amino-4-hydroxybenzoic acid; 2,3,4-trihydroxybenzoic acid; 4-nitrobenzoic acid; 4-methoxybenzoic acid; 4-fluorobenzoic acid (4-fluorobenzoic acid); 3,4-dimethoxybenzoic acid; 2,3-dihydroxybenzoic acid; 2,4-dihydroxybenzoic acid; 2,5-dihydroxybenzoic acid (2,5-dihydroxybenzoic acid); 4-hydroxybenzenesulfonic acid; Terephthalic acid; 3,4-dihydroxyphenylacetic acid; 3,4-dihydroxyphenylacetic acid; D - (-) - tartaric acid (D - (-) - tartaric acid); Dihydroxyfumaric acid hydrate; 2-pyrazinesulfonic acid; 4,4'-sulfonyldibenzoic acid; 5-hydroxynicotinic acid; And 4-hydroxy-5-methyl-4-cyclopentene-1,3-dione monohydrate. And to a pharmaceutical composition for preventing or treating obesity, which comprises a catheter derivative or a salt thereof as an active ingredient.

 The composition of the present invention may further comprise at least one pharmaceutically acceptable carrier in addition to the above-described effective ingredients for administration. Pharmaceutically acceptable carriers include but are not limited to saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, lactose, dextrose, sucrose, sorbitol, mannitol, starch, Alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, One or more components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Furthermore, it can be suitably formulated according to the ingredients, using methods disclosed in the art, or as disclosed in Remington's Pharmaceutical Sciences (recent edition), Mack Publishing Company, Easton PA.

The composition of the present invention can be administered orally or parenterally (for example, intravenously, subcutaneously, intraperitoneally or topically) according to the desired method.

The pharmaceutical compositions of the present invention may be formulated for oral administration, for example as tablets, troches, lozenges, aqueous or oily suspensions, pharmaceutical powders or granules, emulsions, hard or soft capsules, syrups or elixirs elixirs. Binders such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose or gelatin for formulation in tablets, capsules and the like; Excipients such as dicalcium phosphate; Disintegrating agents such as corn starch or sweet potato starch; Lubricating oil such as magnesium stearate, calcium stearate, sodium stearyl fumarate or polyethylene glycol wax. In the case of a capsule formulation, in addition to the above-mentioned substances, a liquid carrier such as fatty oil is contained.

In addition, the pharmaceutical composition of the present invention may be administered parenterally, and parenteral administration may be by subcutaneous injection, intravenous injection, intramuscular injection, or intramuscular injection. The dosage varies depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and disease severity.

The composition of the present invention can be used alone or in combination with methods for the prevention or treatment of anti-obesity or using surgery, hormone therapy, drug therapy and biological response modifiers.

The present invention also relates to a method of preventing or treating obesity of any one selected from the group consisting of a light path or a soft capsule containing the pharmaceutical composition, a tablet, a sugar solution, a film coated tablet, a tablet, a tablet, a powder, a suspension, a syrup, ≪ / RTI >

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited thereto.

Example  One. Protocatechuic acid  Lipase inhibitory activity of derivatives lipase inhibition  acitivity analysis

In the present invention, an anti-obesity substance, 3,4-dihydroxybenzoic acid (PCA), was isolated and purified from the juice of pine needle, and a substance having excellent anti-obesity effect was selected through chemical and structural changes and substitution. Or for the treatment, prevention, amelioration or treatment of a disease or condition.

Therefore, in this Example, the inhibitory activity of lipase, which is a lipolytic enzyme that can inhibit digestion and absorption of lipid and reduce energy intake through the stomach, is analyzed.

[Experimental Method]

30 μl of a lipase buffer (10 mM MOPS, 1 mM EDTA, pH 6.8) was added to 850 μl of Tris buffer (100 mM Tris-HCl, 5 mM CaCl 2, pH 7.0), and 0.3 mg of porcine pancreatic lipase (Porcine pancreatic lipase). 100 μl of the sample was added to the enzyme solution, followed by reaction at 37 ° C for 15 minutes. After the reaction mixture was added to 10 mM p -NPB (Sigma Chemical Co. ) 20㎕ was further reacted at 37 ℃ for 15 minutes. p - nitrophenyl butyrate (Nitrophenyl butyrate; p -NPB) a p - nitrophenol (p -nitrophenol) in the absorbance was measured for the degree of hydrolysis decomposition at 400nm. Lipase (Lipase) inhibitory activity (%) was calculated according to the following equation (1).

(1) Lipase inhibitory activity (%) = [1- (BC) / A] 100

(A is the absorbance without the addition of the sample, B is the absorbance with addition of the sample, and C is the absorbance without addition of the enzyme)

(One) Protocatechuic acid  Derivative Of a hydroxy group  Analysis of lipase inhibitory activity by position and number

Protocatecuic acid (PCA) is a structure with two carboxyl groups (-C (= O) OH) and two hydroxyl groups (-OH) instead of hydrogen in the benzene ring. However, lipase inhibition activity of the protocatechuic acid derivatives was confirmed by the functional groups.

2,3-dihydroxybenzoic acid (Compound 2), 2,4-dihydroxybenzoic acid (compound 3) containing two hydroxyl groups (-OH) ), 2,3,4-trihydroxybenzoic acid (2,3,4-trihydroxybenzoic acid) containing three 2,5-dihydroxybenzoic acid (Compound 4) and a hydroxyl group (-OH) -trihydroxybenzoic acid (Compound 5) was confirmed, and the lipase inhibiting activities of the compounds were almost similar, and it was confirmed that the compounds were active regardless of the number and positions of hydroxyl groups (-OH).

In addition, experiments were performed to investigate whether the side chains in place of the hydroxyl groups (-OH) located at positions 3 and 4 of the above protocatechuic acid affect the lipase inhibitory activity. 3-amino-4-hydroxybenzoic acid (Compound 6) having an amino group instead of a hydroxyl group (-OH) at position 3 and a 3-amino-4-hydroxybenzoic acid 4-Nitrobenzoic acid (Compound 4) containing a nitro group (-NO ₂ ), a methoxy group (-OCH ₃ ) or a fluoro group (-F) 7), 4-Methoxybenzoic acid (Compound 8) or 4-Fluorobenzoic acid (Compound 9) was confirmed to have lipase inhibitory activity. 3,4-Dimethoxybenzoic acid (Compound 10) in which all of the positions 3 and 4 were substituted with a methoxy group (-OCH ₃ ) showed lipase inhibitory activity. The results showed that the side chains were also inhibitory in various substituted derivatives. Therefore, it was confirmed that when a carboxyl group (-C (= O) OH) was present, lipase inhibition activity was not affected by the substituent of the side chain (Table 1 and FIG. 1).

Table 1. Structure and lipase inhibitory activity of protocatechuic acid derivatives (Compound 1 to Compound 10)

(2) Protocatechuic acid  Analysis of lipase inhibitory activity depending on the presence or absence of the carboxyl group (-C (= O) OH) of the derivative

Protocatechuic acid (Compound 1) and 3,4-dihydroxyphenylacetic acid (Compound 11) have a structure in which a carboxyl group (-C (= O) OH) is bonded to a benzene ring Indicating lipase inhibitory activity.

In the case of 4-hydroxybenzenesulfonic acid (Compound 12) having a structure free from a carboxyl group (-C (= O) OH), a sulfonic acid group group), confirming the lipase inhibitory activity.

In addition, terephthalic acid (13) and 4,4'-sulfonyldibenzoic acid (14) having two carboxyl groups (-C (= O) OH) also have lipase inhibitory activity 5-methyl-4-cyclopentene-1,3-dione monohydrate which does not contain both a carboxyl group (-C (= O) OH) and a sulfonic acid group, 4-cyclopentene-1,3-dione monohydrate (Compound 15) also showed lipase inhibitory activity.

However, when salicylamide (Compound 16), Protocatechuic acid ethyl ester (Compound 17), 4-hydroxybenzaldehyde (Compound 16) substituted with other functional groups instead of the carboxyl group (-C 4-Hydroxybenzaldehyde (compound 18), pyrazinecarboxamide (compound 19) and 3,4,5-trihydroxybenzamide (compound 20) showed no lipase inhibitory activity. Therefore, it was determined that the presence of the carboxyl group (-C (= O) OH) influences the lipase inhibitory activity (Table 2 and FIG. 2).

Table 2. Structure and lipase inhibitory activity of protocatechuic acid derivatives (Compound 11 to Compound 20)

(3) Protocatechuic acid ( protocatechuic acid Analysis of Lipase Inhibitory Activity by the Presence of the Benzene Ring Structure of the Derivative

Hydroxynicotinic acid (Compound 21) in order to confirm the lipase inhibitory activity according to the presence of the benzene ring structure, the lipase inhibitory activity of 5-hydroxynicotinic acid (Compound 21) was confirmed. The 5-hydroxynicotinic acid has no benzene ring structure and has a structure containing a carboxyl group (-C (= O) OH), and exhibits lipase inhibitory activity. Then, D - (-) - tartaric acid (Compound 22) and dihydroxyfumaric acid hydrate (Compound 23), which have a carboxylic acid group but not a benzene ring compound, have a lipase inhibitory activity . From the above results, it was judged that the benzene ring structure did not affect the lipase inhibitory activity.

Further, by confirming the inhibitory activity of 2-pyrazinesulfonic acid (Compound 24), which is not a benzene ring structure but contains a sulfonic acid group, it is possible to obtain a sulfonic acid (Table 3 and Fig. 3). ≪ tb > < TABLE >

Table 3. Structure and lipase inhibitory activity of protocatechuic acid derivatives without benzene ring

Claims (7)

3-amino-4-hydroxybenzoic acid; 2,3,4-trihydroxybenzoic acid; 4-nitrobenzoic acid; 4-methoxybenzoic acid; 4-fluorobenzoic acid (4-fluorobenzoic acid); 3,4-dimethoxybenzoic acid; 2,3-dihydroxybenzoic acid; 2,4-dihydroxybenzoic acid; 2,5-dihydroxybenzoic acid (2,5-dihydroxybenzoic acid); 4-hydroxybenzenesulfonic acid; Terephthalic acid; 3,4-dihydroxyphenylacetic acid; 3,4-dihydroxyphenylacetic acid; D - (-) - tartaric acid (D - (-) - tartaric acid); Dihydroxyfumaric acid hydrate; 2-pyrazinesulfonic acid; And 4,4'-sulfonyldibenzoic acid; 5-hydroxynicotinic acid; And 4-hydroxy-5-methyl-4-cyclopentene-1,3-dione monohydrate. A health food composition for preventing or ameliorating obesity comprising a catheteric acid derivative or a salt thereof as an active ingredient. The health food composition for preventing or ameliorating obesity according to claim 1, wherein the protocatechuic acid derivative inhibits lipase activity. The health food composition according to claim 1, wherein the health food composition further comprises at least one selected from the group consisting of flavorings, natural carbohydrates, nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, , a pH adjusting agent, a stabilizer, a preservative, a glycerin, an alcohol, and a carbonating agent used in a carbonated beverage. 3-amino-4-hydroxybenzoic acid; 2,3,4-trihydroxybenzoic acid; 4-nitrobenzoic acid; 4-methoxybenzoic acid; 4-fluorobenzoic acid (4-fluorobenzoic acid); 3,4-dimethoxybenzoic acid; 2,3-dihydroxybenzoic acid; 2,4-dihydroxybenzoic acid; 2,5-dihydroxybenzoic acid (2,5-dihydroxybenzoic acid); 4-hydroxybenzenesulfonic acid; Terephthalic acid; 3,4-dihydroxyphenylacetic acid; 3,4-dihydroxyphenylacetic acid; D - (-) - tartaric acid (D - (-) - tartaric acid); Dihydroxyfumaric acid hydrate; 2-pyrazinesulfonic acid; 4,4'-sulfonyldibenzoic acid; 5-hydroxynicotinic acid; And 4-hydroxy-5-methyl-4-cyclopentene-1,3-dione monohydrate. A pharmaceutical composition for preventing or treating obesity, which comprises a catheteric acid derivative or a salt thereof as an active ingredient. The pharmaceutical composition according to claim 4, wherein the active ingredient is at least one selected from the group consisting of pharmaceutically acceptable salt water, sterilized water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, lactose, dextrose, sucrose, sorbitol, mannitol, But are not limited to, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, Wherein the pharmaceutical composition further comprises at least one carrier selected from the group consisting of magnesium, magnesium and mineral oil. A pharmaceutical preparation for preventing or treating obesity comprising the pharmaceutical composition of claim 4. The pharmaceutical preparation according to claim 6, wherein the pharmaceutical preparation has any one form selected from the group consisting of a light path or a soft capsule, a tablet, a sugar solution, a film coated tablet, a tablet, a tablet, a powder, a suspension, a syrup, A pharmaceutical preparation for preventing or treating obesity.

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