KR20150101164A - Composition comprising monascus pigment derivative for preventing, alleviating or treating obesity - Google Patents

Abstract

The present invention relates to a composition comprising a monascus pigment derivative for preventing, improving or treating obesity. According to the present invention, the composition can effectively prevent, improve or treat obesity by inhibiting an effect of weight gain.

Description

[0001] The present invention relates to a composition for preventing, ameliorating or treating obesity including a monascus pigment derivative,

The present invention relates to a composition for preventing, improving or treating obesity comprising a Monascus pigment derivative.

Recently, the improvement of living standards due to economic development improves the hygiene environment and frequent instant food intake and eating habits change the eating habits of excessive calories. However, the changes in dietary habits of the modern people are due to lack of exercise and the like. Obesity has been reported to cause breast cancer, uterine cancer and colon cancer as well as adult diseases such as hypertension, diabetes, hyperlipidemia, coronary artery disease and the like due to persistent obesity as well as external problems. It is treated as one.

Current treatments for treating obesity include drugs that act on the central nervous system, affect appetite, and drugs that act on the gastrointestinal tract to inhibit absorption. However, these drugs may cause side effects such as primary pulmonary hypertension or heart valve lesion, and problems such as decreased blood pressure or lactic acidosis may occur, resulting in problems such as heart failure and renal disease.

On the other hand, the Monascus pigment is a safety-proven pigment used for edible purposes, and is produced by a microorganism of the genus Monascus sp. Monasukus pigment has been used for fermentation of traditional foods such as Hongju and Hongdubu in East Asian countries including Korea, China, Japan and Indonesia since ancient times.

The Monasukus pigment is known to have a high heat resistance, a broad pH range, and good tenacity and a mixture of more than 10 components.

Monacin, ankaflavin, orange monascus, monascorubrin, rubropunctatin, red bean monascorubramine, and monocarbromine, which are the major components of the monascus pigment, Rubropunctamine, etc. These dye components have different solubility characteristics in various solvents and are known to have different production ratios and yields depending on the strain used, nutritive substrate, and culture conditions. (Chen & Johns, Appl . Microbiol. Biotechnol . , 40: 132-138, 1993).

Discloses Monacus pigment derivatives in KR10-2004-0082143 and KR 10-2009-0033187. KR10-2004-0082143 discloses a lipase-inhibiting activity of an amino acid derivative of a Monascus pigment. However, KR10-2004-0082143 discloses only the inhibitory effect of L-tryptophan, D-tyrosine and L-leucine derivatives of the Monascus pigment on the lipolytic enzyme activity, and discloses a substantial effect of inhibiting the weight gain of monacercastinosin derivatives It is not. In addition, KR 10-2009-0033187 discloses an effect of inhibiting lipid progenitor cell differentiation by a Monascus pigmented amine derivative, but does not disclose a substantial effect of inhibiting the weight gain of a monascus muscle threonine derivative.

Korean Patent Application No. 2004-0082143 Korean Patent Application No. 2009-0033187

Chen & Johns, Appl. Microbiol. Biotechnol., 40: 132-138, 1993

The object of the present application is to prevent or treat obesity of monacercastinosin derivatives.

The present invention provides a pharmaceutical composition for preventing or treating obesity comprising a compound of the following formula

[Chemical Formula 1]

Wherein, R is C _{1 -12} alkyl, C _2-12 alkenyl, C _2-12 alkynyl is.

The present application also provides a food composition for preventing or improving obesity comprising the compound of formula (1).

According to the present application, the effect of weight gain can be suppressed, effectively preventing, improving or treating obesity.

Figure 1 shows the change of color due to the generation of monacercastrin derivative.
Fig. 2 shows HPLC (Fig. 2A) and LC-MS (Fig. 2B) performance results of monacercastedonin derivatives.
Figure 3 shows the effect of inhibiting the weight gain of monacasterthreonine derivatives in mice. (●: General Diet (ND), ○: High Fat Diet (HFD), ▼: HFD + Water (DW), Δ: HFD + Threonine Derivative G / mouse / day, HFD + threonine derivative 0.05 mg / g-mouse / day, HFD + threonine derivative 0.1 mg /
Fig. 4 shows the results of comparing the effects of monacasterthrethanolamine derivatives (TEA derivatives) and tryptophan derivatives (Thr derivatives) against weight gain in accordance with the present application.

The present invention provides a pharmaceutical composition for preventing or treating obesity comprising a compound represented by the following formula (1). The present application also provides a food composition for preventing or improving obesity comprising a compound of formula (1):

[Chemical Formula 1]

Wherein, R is C _{1 -12} alkyl, C _2-12 alkenyl, C _2-12 alkynyl is.

As used herein, the term " alkyl " generally refers to straight and branched saturated hydrocarbon groups having the indicated number of carbon atoms (e.g., 1 to 12 carbon atoms). Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl, t- butyl, pent- Methylbut-2-yl, 2,2,2-trimethylet-1-yl, n-hexyl, n-heptyl and n-octyl, and the like. The term " alkenyl " and " alkynyl " in the present application refers to an alkyl group containing at least one double bond and at least one triple bond. Examples of alkenyl groups include, but are not limited to ethenyl, 1-propen-1-yl, 1-propen-2-yl, 2-propen-1-yl, allyl, and the like. Examples of the alkynyl group include, without limitation, ethynyl, 2-propynyl, 2-butynyl, 1,3-butadinyl and the like. Alkyl, alkenyl, or alkynyl may be attached to a parent group or substrate at any ring atom provided that attachment does not violate the atomic requirement. Likewise, an alkyl, alkenyl or alkynyl group may contain one or more non-hydrogen substituents if the attachment does not violate the valence requirements.

In one embodiment, R is a C _{1 -8} alkyl, C _{2 -8} can be alkenyl or C _{2 -8} alkynyl imidazol.

In one embodiment, R can be C _{4 -8} alkyl, C _{4 -8} alkenyl or C _{4 -8} alkynyl.

In one embodiment, R can be pent-1-yl or n-heptyl.

The compound of Formula 1 of the present application may be prepared by reacting a monacercastrin derivative with a compound of the following Formula 2, which is a Monascus pigment orange, with threonine of Formula 3:

(2)

(3)

Wherein, R is C _{1 -12} alkyl, C _2-12 alkenyl, C _2-12 alkynyl is.

Specifically, the monacaster threonine derivative can be prepared by reacting the monacus orange dye of Formula 2 and the threonine of Formula 3 obtained from the microorganism of the genus Monascus. Although not limited thereto, the compound of Formula 1 may be prepared by reacting a Monascus pigment orange and threonine at room temperature for 30 minutes to 1 hour.

Here, the weight ratio of monascus pigment orange and threonine can be, for example, from 7: 1 to 1: 1, from 5: 1 to 1: 1, or from 3: 1 to 1:

The method for producing the Monascus color orange dye is not particularly limited. For example, a Monascus crustacean microorganism can be cultured in a conventional fermentation medium to obtain a Monascus currant orange pigment. The type of microorganism of the genus Monascus is not limited, and for example, KCCM 10093 (Monascus sp. KCCM 10093) can be used.

More specifically, after adding ethyl acetate to the microbial fermentation broth and adsorbing it on silica gel, a solvent is added to the silica gel-dye mixture, followed by washing and drying to obtain a powdered monacus orange pigment.

When a threonine derivative is produced by the above reaction, the color of the Monascus pigment changes from orange to red. From this color change, the threonine derivative compound can be visually confirmed.

The compound of formula (1) according to the present application has an effect of inhibiting weight gain, and thus can be usefully used for preventing, ameliorating or treating obesity.

For example, when the compound of Chemical Formula 1 is used as a medicine, the pharmaceutical composition containing the compound of Chemical Formula 1 may contain one or more pharmaceutically acceptable carriers in addition to the active ingredient. "Pharmaceutically acceptable carrier" means a known pharmaceutical excipient which is useful when formulating pharmaceutically active compounds for administration and which is substantially non-toxic and non-sensitive under the conditions of use. The exact ratio of such excipients is determined by standard pharmaceutical practice, as well as the solubility and chemical properties of the active compound, the route of administration chosen.

The pharmaceutical composition of the present application can be formulated into a form suitable for a desired administration method using suitable and physiologically acceptable excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, lubricants, have.

The pharmaceutical composition may be formulated in the form of tablets, capsules, pills, granules, powders, injections or solutions, though it is not limited thereto.

Formulations of the pharmaceutical compositions and pharmaceutically acceptable carriers may be appropriately selected according to techniques known in the art, for example, see: Urquhart et al., Lancet, 16: 367, 1980 ]; [Lieberman et al., PHARMACEUTICAL DOSAGE FORMS-DISPERSE SYSTEMS, 2nd ed., Vol. 3, 1998]; [Ansel et al., PHARMACEUTICAL DOSAGE FORMS & DRUG DELIVERY SYSTEMS, 7th ed., 2000]; [Martindale, THE EXTRA PHARMACOPEIA, 31st ed.]; [Remington ' s PHARMACEUTICAL SCIENCES, 16th-20th editions]; [THE PHARMACOLOGICAL BASIS OF THERAPEUTICS, Goodman and Gilman, eds., 9th ed., 1996]; [Wilson and Gisvolds' TEXTBOOK OF ORGANIC MEDICINAL AND PHARMACEUTICAL CHEMISTRY, Delgado and Remers, eds., 10th ed., 1998]. The principles of formulating pharmaceutical compositions are also described, for example, in Platt, Clin Lab Med, 7: 289-99, 1987; [Aulton, PHARMACEUTICS: THE SCIENCE OF DOSAGE FORM DESIGN, Churchill Livingstone, NY, 1988]; [EXTEMPORANEOUS ORAL LIQUID DOSAGE PREPARATIONS, CSHP, 1998], ["Drug Dosage," J Kans Med Soc, 70 (1): 30-32, 1969).

An effective amount of the compound of formula (I) of the pharmaceutical composition of the present application means an amount required for the treatment of the disease. Accordingly, the present invention is not limited to the particular type of the disease, the severity of the disease, the kind and amount of the active ingredient and other ingredients contained in the composition, the type of formulation and the patient's age, body weight, general health status, sex and diet, Rate of administration, duration of treatment, concurrent medication, and the like. For example, in the case of an adult, the compound of the formula (1) can be administered in a total dose of 50 to 3000 mg / kg once or several times a day, for example, in a total dose of 100 to 1000 mg / can do.

In one embodiment, when the compound of formula (1) is used as a food, the food composition comprising the compound of formula (1) may contain, in addition to the compound of formula (1), monosaccharides such as glucose and fructose; Disaccharides such as maltose, sucrose; Or polysaccharides such as dextrin, cyclodextrin may be added. Sugar alcohols such as xylitol, sorbitol and erythritol; Sweeteners such as taurine, stevia extract and the like may be further added, and further, other food additives may be added.

The food composition of the present application can be formulated in the same manner as the above-mentioned pharmaceutical composition and used as a functional food or added to various foods.

Foods to which the composition of the present application can be added include, for example, beverages, meat, chocolates, foods, confectionery, pizza, ramie noodles, other noodles, gums, candy, ice cream, alcoholic beverages, And dairy products.

As a result of the weight gain test of the compound of Chemical Formula 1 of the present application, it was confirmed that the mice that were fed with the high fat diet had a significant weight gain inhibitory effect. The weight gain suppression effect was dependent on the concentration of the compound of formula (1). (Example 1). It was also found that the monacasterthreonine derivative of the formula (1) had an excellent weight gain inhibitory effect as compared with the monacastus triethanolamine derivative and the monacastus tryptophan derivative, which are known in the art (Example 2). In particular, in the case of threonine derivatives, threonine derivatives (Threonine (Thr)) synthesized with a Monacus orange pigment and a threonine derivative (Thr) are considered harmless and harmless to human body, And can be used as a food additive of the present invention.

The advantages and features of the present application, and how to accomplish them, will become apparent with reference to the embodiments described in detail below. However, the present application is not limited to the embodiments disclosed below but may be embodied in various different forms, and these embodiments are provided so that the disclosure of the present application is complete.

[Example]

[Preparation Example 1] Synthesis of amino acid derivative of Monascus pigment

Manufacturing example  1. 1 Monaskus  Production of orange pigment through culture of strain

Monascus sp. J101 (KCCM 10093) was plated on a slope culture medium and platelets were treated with 75 ml of Mizutani medium (5.0% glucose, 2.0% Bacto peptone, 0.8% KH ₂ PO ₄ , 0.2% CH ₃ COOH, 0.1% NaCl, 0.05% MgSO ₄ .7H ₂ O), and then cultured for 48 hours at 30 ° C in a rotary shaker at 150 rpm to prepare a first seed culture , And 5 ml of the resulting solution was cultured under the same conditions as the first seed culture for 24 hours to carry out the second seed culture. Then, 3 L fermentation medium (5% glucose, 0.3% NH ₄ NO ₃ , 0.1% NH ₂ PO ₄ , 0.05% MgSO ₄ .7H ₂ O, 0.05% KCl and 0.001% FeSO ₄ · 7H ₂ O), and the cells were cultured for 120 hours at 30 ° C., 500 rpm, and 1 vvm air supply rate. The orange pigment content was measured at 470 nm using a spectrophotometer, and the final yield of the fermentation broth was 300 OD units. The resulting fermentation broth was filtered, and the filtrate was discarded and only the residue was collected.

The residue thus obtained was added to a 4000 ml glass vessel, about 3000 ml of ethyl acetate was added thereto, and the mixture was extracted on a reciprocal shaker for 24 hours. Only the filtrate was collected through filtration. The extract which was filtered with an evaporator was concentrated to an amount of about 50 ml. 50 ml of the concentrated dye solution was taken out and adsorbed on 10 g of silica gel and dried in a rotary evaporator to remove ethyl acetate. Hexane was added to 50 ml of the silica gel-dye mixture, washed and dried, and then 30 ml of ethylacetate was added thereto to extract an orange dye and then dried to obtain an orange dye in the form of a final powdered powder.

Manufacturing example  1. 2 Monaskus  Synthesis of threonine derivatives of pigments

The orange pigment of Preparation 1.1 dissolved in ethanol and the threonine dissolved in distilled water were dissolved in 10% Mixed with phosphate buffer (pH 9.7), and the mixture incubated for one hour with stirring. Thereafter, the potassium phosphate buffer was removed to obtain a monascus pigmented threonine derivative.

The obtained threonine derivative can be confirmed by observing the color change visually. When an amino acid is introduced into the monascus color orange pigment, the orange color changes from orange to red as the oxygen molecule of the orange color is substituted with nitrogen of the amino acid (Fig. 1).

HPLC (HP-110, manufactured by Hitachi, Ltd.) under GOLD C ₁₈ column (Hypersil, 250 x 4.6 mm, 5 um) for 40 minutes at a rate of 0.8 ml / min under gradient conditions (100: 0 to 30:70, distilled water: methanol) USA) was performed to confirm the purity of the obtained threonine derivative (Fig. 2A).

In order to identify monacsus pigment threonine derivatives, the molecular weight was measured by LC-MS analysis. Using a UPLC LTQ Orbitrap XL (Thermofix Scientific, USA) with an ACQUITY UPLC BEH C18 column (2.1 x 50 mm, 1.7 um; Waters) at a rate of 0.3 ml / 0 to 30:70, distilled water / 60% MeOH + 40% acetonitrile) at 240 nm. Mass analysis was performed in ESI-positive mode. The spray voltage was 5 kV. The injection rates of nitrogen sheath gas and auxiliary gas were 50 and 5 (arbitrary units). The capillary voltage (V), tube lens voltage (V), and capillary temperature (占 폚) were maintained at 35 V, 100 V, and 370 ° C, respectively.

As a result, C ₅ H ₁₁ and C ₇ H ₁₅ of R of the L-threonine derivative were confirmed by m / z values of 456 and 484, respectively (Fig. 2B).

[Example 1] Inhibitory effect of Monacsuk thiourethrin (Threonine; Thr) derivatives on body weight gain

In vivo (in In order to confirm the anti-obesity effect of the monacsus pigment Thr derivative synthesized according to Preparation Example 1, a weight increase experiment was carried out using C57BL / 6 mouse.

As shown in the following Table 1, the mouse group to which the high fat diet was given was used as a control group, and the mouse group to which the monascus pigmented threonine derivative was orally administered together with the high fat diet group was classified into experimental groups.

High-fat diets were fed to mice with Harlan Teklad (60% of calories in fat) and the diet fed a rodent chow.

The mice in the experimental group were orally administered the pigment derivatives at a concentration of 0.01 mg / g-mouse, 0.05 mg / g-mouse and 0.1 mg / g-mouse daily.

Mouse Group and Dietary Serving Table group Diet Control group HFD Provides high fat diet ND A generic diet HFD (DW) Provides only high-fat diets and water without Thr derivatives Experimental group MTD0.01 High fat diet and 0.01 mg Thr derivatives MTD0.05 High fat diet and 0.05 mg Thr derivatives MTD0.1 High fat diet and 0.1 mg Thr derivatives

(HFD = high fat diet, ND = normal diet)

Six mice per group were raised in each cage for 12 weeks at the Laboratory Animal Research Center of Yonsei University. Dietary intake and body weight of the mice were measured twice weekly using an electronic balance. After 12 weeks, changes in body weight and dietary consumption of the control and experimental groups were analyzed.

As a result, the group fed the high fat diet increased 2.8 times in body weight compared to the group fed the normal diet. On the other hand, the group fed with threonine derivatives showed a decrease in weight gain compared to the group fed high fat diets. In particular, mice of the MTD0.1 group showed about 43% less weight gain than the high fat diet group (FIG. 3).

[Example 2] Comparison of effect of inhibiting weight gain of Monascus pigment derivative

In order to compare the effects of various monascus pigment derivatives and the threonine derivatives of the present invention on the inhibition of weight gain, it is known that Monacus pigmentary triethanolamine (TEA) derivatives, which are known to have lipolytic enzyme activity, and Monacus pigment Tryptophan (Trp) derivatives were selected and weight gain experiments were carried out under the same conditions as in Example 1, except that the mice of each experimental group were orally administered a derivative of 0.5 mg / g-mouse concentration.

The triethanolamine derivative can be prepared in the same manner as in Production Example 1.2, except that the triethanolamine is a liquid and the triethanolamine is dissolved in water. The tryptophan derivative can be prepared in the same manner as in Production Example 1.2, except that tryptophan is used instead of threonine.

As a result of the weight gain test, the weights of 10.63 g, 8.33 g, 8.86 g and 6.3 g of the control, Trp derivative, TEA derivative and Thr derivative increased, respectively. The body weight gain inhibitory activity of the Trp derivative was 21.6%, the TEA derivative had the weight gain inhibitory activity of 16.7%, and the Thr derivative was 40.7% of the control group. That is, the Thr derivative has a weight gain inhibitory activity (19.1%) higher than the Trp derivative and 24% higher than the TEA derivative (FIG. 4).

This means that the threonine derivative of the present application has a far superior weight gain inhibitory effect as compared with other amines and amino acid derivatives.

Claims (7)

A pharmaceutical composition for preventing or treating obesity comprising a compound of the following formula (1):
[Chemical Formula 1]

Wherein, R is C _{1 -12} alkyl, C _2-12 alkenyl, C _2-12 alkynyl is.
The method of claim 1, wherein, R is a C _{1 -8} alkyl, C ₂ alkenyl or C _{2 -8 -8} alkynyl obesity prevention or treatment a pharmaceutical composition for.
The pharmaceutical composition for preventing or treating obesity according to claim 1, wherein R is C _{4 -8} alkyl, C _{4 -8} alkenyl or C _{4 -8} alkynyl.
The pharmaceutical composition for preventing or treating obesity according to claim 1, further comprising a pharmaceutically acceptable carrier.
A food composition for preventing or improving obesity comprising a compound of formula
[Chemical Formula 1]

Wherein, R is C _{1 -12} alkyl, C _2-12 alkenyl, C _2-12 alkynyl is.
The method of claim 1, wherein, R is a C _{1 -8} alkyl, C ₂ alkenyl or C _{2 -8 -8} alkynyl a food composition for preventing or improving obesity.
The food composition for preventing or improving obesity according to claim 1, wherein R is C _{4 -8} alkyl, C _{4 -8} alkenyl or C _{4 -8} alkynyl.

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