KR101016787B1 - The diet composition containing the extract of Mori folium, fraction thereof or compounds isolated therefrom for prevention or improvement of depression or anxiety, or for inhibition of appetite as an active ingredient - Google Patents

Abstract

The present invention relates to a leaf extract, a dichloromethane fraction thereof or an active fraction isolated therefrom or perpurin-7 dimethyl ethyl ester, hydroxy-phephorbide A ethyl ester, phephorbide A ethyl ester, didehydro- rodochlorine ethyl The present invention relates to a food composition for suppressing appetite, preventing and improving depression, or preventing and improving anxiety, containing methyl ester, hydroxy pheophorbide A methyl ester, or pheophorbide A methyl ester or a mixture thereof as an active ingredient. The composition according to the present invention exhibits a strong antagonistic action with Melanin Concentration Hormone receptor subtype-1 (MCH-1), thus exhibiting appetite suppression effect, weight loss effect, etc. It can be usefully used for prevention and improvement, or prevention and improvement of anxiety.

Leaf Extract, MCH, Obesity, Perpurin-7 Dimethyl Ethyl Ester, Pheophorbide A Ethyl Ester, Hydroxy-Peophorbide A Ethyl Ester

Description

Food composition for inhibiting appetite, preventing and improving depression, or preventing and improving anxiety, containing an upper leaf extract, a fraction thereof or a compound separated therefrom as an active ingredient isolated therefrom for prevention or improvement of depression or anxiety, or for inhibition of appetite as an active ingredient}

The present invention relates to a food composition for suppressing appetite, preventing and improving depression, or preventing and improving anxiety based on MCH-1 receptor antagonistic activity containing an upper leaf extract, a fraction thereof or a compound separated therefrom as an active ingredient.

Obesity (obesity) refers to a state in which the excessive accumulation of fat tissue in the body. Each person has a standard weight that suits him or her, which is defined as obesity when there is an increase of more than 120% of the individual's standard weight. ), The mortality rate is minimal when the BMI is 22 kg / m ^{2 in} both adults and men, and the risk of death is increased when the BMI is above 27 kg / m ² . Specifically, when the weight of fat accounts for more than 25% of body fat in men and more than 30% in women [Van der Ploeg LH, Curr Opin Chem Biol. 4 (4), pp 452-460, 2000]. Obesity is also caused by environmental factors such as overingestion of energy and lack of exercise, but also by neuroendocrine causes, drug causes, genetic factors and biochemical reactions. Obesity is not just a person's greed or laziness, but it causes long-term management by causing various chronic degenerative diseases such as hypertension, arteriosclerosis, coronary artery disease, type 2 diabetes, fatty liver, hyperlipidemia, degenerative arthritis, and some cancer diseases. It is one of chronic diseases. In particular, in Asians, obesity management is more important than other Westerners because their body mass index is at least abdominal obesity, and the sensitivity of atherosclerosis-related diseases such as hypertension, diabetes, and hyperlipidemia is high.

As the severity of obesity is added, many researches and developments on materials and products for the prevention and treatment of obesity have been made, but currently developed anti-obesity drugs have side effects.

For example, there are two types of drugs that are approved for long-term use by the US Food and Drug Administration (FDA) as obesity drugs. However, recent reports show that sibutramine (reductil) acts on the hypothalamus to increase satiety, inhibit eating, and increase energy consumption, resulting in an average weight loss of 6.1 kg per year. It is known, but it is known to increase blood pressure by 2 mmHg and increase pulse rate 3-6 times per minute.

In addition, Xenical (orlistat), an over-the-counter anti-obesity drug, induces energy loss of 200 Kcal by inducing 30% of dietary fat without digesting or absorbing it. It is known that the average weight loss effect is 8.8 kg. However, side effects may include secretion of gas discharge, oily stool, etc., which impedes continuous medication administration and requires doctor's prescription [Fujioka K et al., Diabetes Obes. Metab., 2 (3), pp 175-187, 2000; Leung WYS et al., Clinical Therapeutics, 25 (1), pp 58-80, 2003; McMahon FG et al., Arch. Int. Med., 160 (14), pp 2185-2191, 2000]. Therefore, the development of new functional materials without side effects for the improvement and prevention of obesity is constantly required.

Meanwhile, as the role of food intake behavior of melanin-concentrating hormone (MCH) consisting of 19 amino acids has been revealed, a new treatment for treating obesity by antagonizing the action of MCH has recently attracted attention.

Antagonists that can modulate MCH-1 receptors, one of the G-protein coupled receptors (GPCRs) that mediate the effects of MCH, have been reported to be useful for treating depression, anxiety, anxiety, etc. [B] . Borowsky et al., Nature Medicine, 8 (8), 825-30, 2002]. MCH was initially known as a factor controlling color change in tibial fish, which consists of 17 amino acids in fish and 19 amino acids in mammals, and these sequences are identical in rodents, rabbits, and humans. Intracellular signal transduction through GPCR. Two types of receptors, MCH-1 (SLC-1, GPR24) and MCH-2 (SLT, GPRv17), have been reported. MCH-1, known in 1999, consists of 353 amino acids and is reported to have typical GPCR characteristics, with about 35% similarity to the somatostatin receptor, and the gene is known to be located at chromosome 22q13.3. [P. Pssios et al., Trends Endocrinol Metab. 14 (5), 243-8, 2003]. The study of MCH function in animal models showed that food intake increased in MCH-treated mice and weight loss due to decreased feeding and increased metabolic rate in mice lacking MCH [D. Qu., Et al., Nature, 380 (6571), 243-7, 1996]. The effect of MCH on eating behavior is known to be due to antagonism of MCH-1 receptors, and it has been reported that administration of MCH to MCH-1 knock-out animals does not stimulate feeding or cause obesity. [A. L. Handlon and H. Zhou, J. Med. Chem. 49, 4017-22, 2006]. Another study of the importance of the MCH-1 system in regulating energy balance against obesity is the experiment using SNAP-7941 as an MCH-1 receptor antagonist. Animals treated with these compounds exhibited significant weight loss and are known to provide additional anxiety and antidepressant effects in addition to the loss of appetite [B. Borowsky et al., Nature Medicine, 8 (8), 825-30, 2002].

According to the results of the series of studies, MCH-1 receptor antagonists can be used not only as an anti-obesity agent but also effective in removing depression and anxiety.

Mori folium is a dried leaf of mulberry and cousin myosin. Its taste is sensitive and its properties are limited. The leaves contain rutin, quercetin, moracetin, beta-sitosterol, campesterol, lupeol, myinositol and inos. It contains theinkosterone and the like. The upper lobe is known to have effects such as nourishing blood (reducing the heat of the blood and reducing moisture), big wind nominal (removing the wind and clearing the eyes), and instructing the abdomen (when taken in powder, eliminating cold sweat). . In addition, it has been recorded that the oriental medicines such as the herbaceous tree have an effect on small thirst along with baekbaekpi and silkworm cocoons. Dohae Herbal Medicine (Medicinal Herb) Dictionary, Younglimsa, pp 545-546, 1998].

However, in relation to the efficacy of the upper lobe, there is no report on the effect of the lobe on the antagonism of the MCH-1 receptor.

Therefore, the inventors of the present invention, while searching for an active substance having excellent MCH-1 receptor antagonism in natural products, the extract of the upper leaf and the compound isolated therefrom have an excellent antagonism against the MCH-1 receptor, leading to appetite suppression and weight loss. The present invention has been found to be useful in the prevention and improvement of obesity and various metabolic disorders and eating disorders resulting therefrom.

SUMMARY OF THE INVENTION An object of the present invention is a food composition for inhibiting appetite, preventing and improving depression, or preventing and improving anxiety, containing an upper leaf extract having excellent antagonistic action against MCH-1 receptor, a fraction thereof or a compound separated therefrom as an active ingredient. To provide.

In order to achieve the above object, the present invention provides an extract of an upper leaf, a fraction thereof, or perpurin-7 dimethyl ethyl ester, hydroxy-phephorbide A ethyl ester, phephorbide A ethyl ester, and didehydro-chlorodochlorine ethyl. Provided is a food composition for suppressing appetite, preventing and improving depression, or preventing and improving anxiety, containing a compound of methyl ester, hydroxy pheophorbide A methyl ester, or pheophorbide A methyl ester as an active ingredient.

Upper leaf extract according to the present invention, fractions thereof, or purified therefrom, purified purpurin-7 dimethyl ethyl ester, hydroxy-phephorbide A ethyl ester, phephorbide A ethyl ester, didehydro-lodochlorine ethylmethyl ester, A composition containing a compound of hydroxy pheophorbide A methyl ester or pheophorbide A methyl ester as an active ingredient exhibits a strong antagonistic action with the MCH-1 receptor, resulting in an appetite suppression effect, weight loss effect and the like. It can be usefully used for suppressing appetite based on 1 receptor antagonistic activity, preventing and improving depression, or preventing and improving anxiety.

Hereinafter, the present invention will be described in detail.

The present invention provides a food composition for suppressing appetite, preventing and improving depression, or preventing and improving anxiety, containing the extract of the leaf as an active ingredient.

The upper leaf extract according to the present invention can be obtained by extracting from the upper leaf or its dried product, and the upper leaf can be used without limitation, such as those grown or commercially available.

The upper leaf extract according to the present invention may use a conventional extraction method such as solvent extraction method, ultrasonic extraction method, filtration method and reflux extraction method, preferably can be prepared by solvent extraction. Specifically, 2 to 200 times, preferably 10 to 30 times, the extraction solvent of the upper leaf volume is added to the dry and fine upper leaves, and extracted at 10 to 30 ° C. for 1 to 20 days, preferably 5 to 10 days, and filtered. After that, the extract may be prepared by concentration under reduced pressure. In this case, water, an organic solvent or a mixture thereof may be used as the extraction solvent. At this time, the organic solvent may be C ₁ -C ₄ alcohol, dichloromethane, ethyl acetate and a mixed solvent thereof, preferably methanol or ethanol.

The present invention also provides a food composition for suppressing appetite, preventing and improving depression, or preventing and improving anxiety, containing the upper fraction as an active ingredient.

The upper leaf fraction according to the present invention may be obtained by suspending the upper leaf extract in distilled water and fractionating it with butanol, dichloromethane or ethyl acetate, and preferably fractionating with dichloromethane. According to a preferred embodiment of the present invention, the upper leaf or its dried product is cooled for 7 days using ethanol, filtered and the filtrate is concentrated under reduced pressure to obtain an ethanol extract, the ethanol extract is suspended in distilled water and then fractions with the same amount of dichloromethane. The upper fractions can be obtained by concentrating under reduced pressure.

Thereafter, the dichloromethane fraction may be further fractionated using hexane or methanol.

Furthermore, the present invention provides for inhibiting appetite, preventing and improving depression, or preventing and improving anxiety, containing the active fraction separated from the upper leaf fraction as an active ingredient.

The active fractions according to the invention can be separated and purified by chromatography. The chromatography is preferably performed 1 to 2 times silica gel column chromatography. Hexane / ethyl acetate (100: 1 to 1: 1) mixed solvent can be used as a mobile phase. At this time, the solvent used is eluted by a gradient gradient elution method of raising the concentration sequentially from non-polar to polar, and the desired active fraction can be obtained by measuring the blood pressure lowering effect of the collected isolate. At this time, the flow rate of the mobile phase is preferably 2 ~ 4 ml / min.

In addition, the present invention is a Purpurin-7 dimethyl ethyl ester (Purpurin-7 dimethyl ethylester) represented by the following formula (1), Pheophorbide A ethylester represented by the formula (2), hydroxy represented by the formula (3) -Hydroxy-pheophorbide A ethylester, Didehydro-rhodochlorin ethylmethylester represented by the formula (4), Hydroxy pheophorbide A methylester represented by the formula (5) -pheophorbide A methylester and Pheophorbide A methylester represented by the formula (6) or a mixture thereof as an active ingredient to suppress appetite, to prevent and improve depression, or to prevent and improve anxiety do.

The active material of Chemical Formulas 1 to 6 may be obtained by the following preparation method:

Adding the upper leaf with water, an organic solvent or a mixture thereof to obtain an upper leaf extract (step 1);

Suspending the upper leaf extract obtained in step 1 in water and fractionating with dichloromethane to obtain a dichloromethane fraction (step 2);

Dichloromethane fractions obtained in step 2 using a mixed solvent of n- hexane and ethyl acetate as a mobile phase to perform a concentration gradient column chromatography to obtain an active fraction (step 3);

Separating the active fraction from the active fraction obtained in step 3 by performing concentration gradient column chromatography again using a mixed solvent of n-hexane and ethyl acetate as a mobile phase (step 4); And

The fraction separated in step 4 is a mixed solvent of n-hexane and ethyl acetate as a mobile phase, but performing a silica gel column chromatography with different concentration gradients to separate the compound of formula 1-6 (step 5).

In the above production method, step 1 is a step of obtaining the upper leaf extract by adding the upper leaf to water, an organic solvent or a mixture thereof.

The organic solvent may be any one selected from the group consisting of C ₁ -C ₄ alcohol, dichloromethane and ethyl acetate, or a mixed solution thereof, preferably methanol or ethanol.

At the time of extraction, an organic solvent of 2 to 200 times the volume of the upper leaf, preferably 10 to 30 times is added, the extract is extracted at 10 to 30 ° C. for 1 to 20 days, preferably 5 to 10 days, filtered and concentrated under reduced pressure. Extracts can be prepared.

Next, step 2 is a step of suspending the upper leaf extract obtained in step 1 in water and fractionating with dichloromethane to obtain a dichloromethane fraction.

At this time, the dichloromethane fraction may be used a method commonly used in the art, for example, the upper leaf ethanol extract obtained in step 1 is suspended in distilled water, fractionated into the same amount of dichloromethane and concentrated under reduced pressure Dichloromethane fractions can be obtained.

Next, step 3 is a step of concentration gradient column chromatography using the dichloromethane fraction obtained in step 2 using a mixed solvent of n- hexane and ethyl acetate as a mobile phase to obtain an active fraction.

In the above method, the active fraction of step 3 may be obtained by performing a concentration gradient silica gel column chromatography on the dichloromethane fraction obtained in step 2 using a mixed solvent of n-hexane and ethyl acetate. At this time, the concentration gradient of the mixed solvent of n-hexane and ethyl acetate is preferably from 100: 1 to 1: 1, it can be obtained by eluting at a rate of 2 ~ 4 ml / min.

Next, step 4 is a step of separating the active fraction by performing concentration gradient column chromatography on the active fraction obtained in step 3 using a mixed solvent of n-hexane and ethyl acetate.

At this time, the third fraction in the first to fourth fractions of the active fraction obtained in step 3 is a mixed solvent of n-hexane and ethyl acetate as a mobile phase and the concentration gradient is 30: 1 to 1: 1 with a Florisil column It is preferable to separate by chromatography, and it is preferable that the flow rate of the said mobile phase is 2-4 ml / min.

Next, in step 5, the fraction separated in step 4 is a mixed solvent of n-hexane and ethyl acetate as a mobile phase, and silica gel column chromatography is performed by varying concentration gradients to separate the compound of Chemical Formula 1-6. Step.

In step 5, the concentration gradient of the mixed solvent of n-hexane and ethyl acetate is 15: 1 to 1: 3 using silica gel column chromatography for the third to sixth fractions of the fractions separated into nine fractions in step 4. It can be obtained by separating the compound of formula 1-6 by eluting with 1, 30: 1 to 1: 1 or 5: 1 to 1: 1. At this time, it is preferable that the flow velocity of the said mobile phase is 2-4 ml / min.

Compounds of Chemical Formulas 1 to 6 isolated through the above steps were obtained by NMR analysis, respectively, for Purpurin-7 dimethyl ethyl ester, ferrophorbide A ethyl ester, hydroxy-phefobide A ethyl ester, and didehydro-chlorodochlorine ethyl. It was identified as methyl ester, hydroxy pheophorbide A methyl ester and pheophorbide A methyl ester.

Upper leaf extract according to the present invention, fractions thereof, or perpurin-7 dimethyl ethyl ester, ferrophorbide A ethyl ester, hydroxy-pheophorbide A ethyl ester, didehydro-rhodochlorine ethylmethyl ester, hydroxy separated therefrom Roxy pheophorbide A methyl ester and pheophorbide A methyl ester showed excellent MCH-1 receptor binding inhibitory activity in MCH-1 receptor binding inhibition experiment (see Table 1 and Table 2). In addition, in the appetite suppression and weight loss effect experiment from the obese mouse model using the upper leaf extract or the compound isolated therefrom, the upper leaf extract or the compound isolated from the weight of the obese mouse compared to the control group Was decreased from 2 days after administration (see FIG. 1 ), and was observed to show a sustained loss of appetite throughout the experiment (see FIG. 2 ). Through this, the upper leaf extract of the present invention or a compound isolated therefrom inhibits appetite by antagonizing MCH-1 receptor action, and decreases body weight by suppressing appetite based on MCH-1 receptor antagonistic activity, preventing and improving depression, or anxiety It can be usefully used for the prevention and improvement of.

The composition of the present invention may contain one or more active ingredients exhibiting the same or similar functions in addition to the above-mentioned leaf extract, fractions thereof, or compounds separated therefrom.

In order to achieve the above effect, the upper leaf extract of the present invention, Perpurin-7 dimethyl ethyl ester, ferrophorbide A ethyl ester, hydroxy-pheophorbide A ethyl ester, didehydro-rhodochlorine ethyl methyl ester, Examples of foods to which hydroxy pheophorbide A methyl ester and pheophorbide A methyl ester or mixtures thereof can be added include, for example, various foods, beverages, snacks, confectionery, gums, ice creams, tea bags, instant foods. Tea, granules, flavors, vitamin complexes, and other health supplements, but are not limited thereto.

Leaf extract of the present invention, Perpurin-7 dimethyl ethyl ester, ferrophorbide A ethyl ester, hydroxy-phephorbide A ethyl ester, didehydro-rhodochlorine ethylmethyl ester, hydroxy perobide A isolated therefrom Methyl esters and pheophorbide A methyl esters or mixtures thereof may be added to the raw materials in the manufacture of foods or mixed appropriately with the cooked foods to prepare the above-mentioned health-promoting foods or beverages, in which case the final foods Or the content of the lettuce extract, Perpurin-7 dimethyl ethyl ester, hydroxy-phephorbide A ethyl ester, phephorbide A ethyl ester, or mixtures thereof, in the beverage may range from 0.01 to 30% by weight, respectively. have.

The health promoting food or beverage of the present invention may further include other medicinal herbs or extracts thereof that are pharmaceutically acceptable to enhance or supplement the desired effect, and representative examples of such herbal medicines are Baekbokryeong, Baekchul, Uiin, Red bean head, neck, cha-chan and the like. The herbal medicine may be used in an amount of 0.01 to 50% by weight based on the total weight of the composition.

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

However, the following Examples, Experimental Examples, and Formulation Examples specifically illustrate the present invention, and the content of the present invention is not limited to Examples, Experimental Examples, and Formulation Examples.

Example 1 Preparation of Upper Leaf Extract

<1-1> Ethanol Extract of Upper Leaf

After cutting the dried upper leaf leaves 1.8 kg into an extraction container and immersed in 100% ethanol 5 times at room temperature, cooled for one week and filtered through a filter paper. The extract was concentrated under reduced pressure at 50 ° C. until the solution was completely evaporated using a concentrator to obtain 370 g of ethanol extract.

<1-2> Preparation of Water Extract of Upper Leaf

300 g of the water extract of the upper leaf was obtained in the same manner as in addition to using water instead of ethanol in the method of <1-1>.

< Example  2> Upper lobe Dichloromethane Fraction  Produce

      The upper leaf ethanol extract (370 g) prepared in Example <1-1> was suspended in 2 L of distilled water, extracted with the same amount of dichloromethane, the dichloromethane layer was separated and concentrated under reduced pressure to give the upper dichloromethane. Fractions (59 g) were obtained and the remaining water layer was lyophilized.

< Example  3> Upper lobe Dichloromethane Fraction Hexane Fraction  Produce

The dichloromethane fraction (57 g) prepared in Example 2 was suspended in 2 L of distilled water, extracted with the same amount of hexane, the hexane layer was separated and concentrated under reduced pressure to obtain a hexane fraction (30 g). Was lyophilized.

< Example  4> Upper lobe Dichloromethane Fraction  Methanol Fraction  Produce

      The dichloromethane fraction (57 g) prepared in Example 2 was suspended in 2 L of distilled water, extracted with the same amount of methanol, the methanol layer was separated and concentrated under reduced pressure to obtain a methanol fraction (32 g), and the remaining water layer Was lyophilized.

< Example  5> Upper lobe Dichloromethane From fractions Active fraction  Produce

The fraction of Example 2 (57 g) was run at a flow rate of 3 ml / min using a mixed solvent (100: 1 to 1: 1) of n-hexane and ethyl acetate, and then subjected to concentration gradient silica gel column chromatography. The total was divided into four fractions (first fraction to fourth fraction). A third fraction of this was obtained as an active fraction.

< Example  6> Upper lobe From active fraction  Preparation of Compound of Formula 1

The active fraction obtained in Example 5 was separated by Florisil column chromatography ( n -hexane: ethyl acetate = 30: 1 to 1: 1 as mobile phase, flow rate 3 ml / min), divided into nine fractions, Silica gel column chromatography ( n -hexane: ethyl acetate = 15: 1 ~ 1: 1) was performed on the 3-4 fractions to separate 44 mg of Perpurin-7 dimethyl ethyl ester of Chemical Formula 1.

¹ H-NMR (300 MHz, Chloroform-d) δ 9.51 (1H, s, H-10), 9.10 (1H, s, H-5), 8.45 (1H, s, H-20), 7.71 (3H, dd, J = 17.9, 11.4 Hz, H-3 ¹ ), 6.16 (1H, d, J = 17.9 Hz, H-3 ² _a ) 6.02 (1H, d, J = 11.4 Hz, H-3 ² _b ), 4.82 (1H, dd, J = 9.0, 2.0 Hz, H-17), 4.43 (1H, q, J = 7.2 Hz, H-18), 4.26 (3H, s, H-15 ³ ), 4.10 (2H, q, J = 7.1 Hz, H-17 ⁴ ), 4.00 (3H, s, H-13 ² ), 3.64 (3H, s, H-12 ¹ ), 3.49 (2H, q, J = 7.6 Hz, H- 8 ¹ ), 3.28 (3H, s, H-2 ¹ ), 3.01 (3H, s, H-7 ¹ ), 2.44 (1H, m, H-17 ¹ _a ), 2.23 (1H, m, H-7 ¹ _b ), 2.10 (2H, m, H-17 ² ), 1.91 (3H, d, J = 7.2 Hz, H-18 ¹ ), 1.20 (3H, t, J = 7.1 Hz, H-17 ⁵ ), 0.00 (2H, br.s, NH)

¹³ C-NMR (125 MHz, Chloroform-d) δ 186.6 (C-15 ¹ ), 174.5 (C-1), 173.2 (C-17 ³ ), 168.0 (C-13 ¹ ), 166.7 (C-15 ² ), 164.3 (C-19), 156.2 (C-9), 155.8 (C-6), 149.1 (C-11), 145.7 (C-8), 138.7 (C-14), 136.7 (C-4) , 136.6 (C-16), 136.3 (C-3), 135.8 (C-7), 131.4 (C-2), 129.7 (C-12), 128.7 (C-3 ¹ ), 122.6 (C-3 ² ), 119.5 (C-13), 106.4 (C-10), 106.3 (C-15), 97.7 (C-5), 93.8 (C-20), 60.5 (C-17 ⁵ ), 53.4 (C-13 ² ), 52.8 (C-17), 52.1 (C-15 ³ ), 49.8 (C-18), 32.0 (C-17 ¹ ), 31.4 (C-17 ² ), 23.3 (C-18 ¹ ), 19.4 (C-8 ¹ ), 17.6 (C-8 ² ), 14.3 (C-17 ⁴ ), 13.1 (C-12 ¹ ), 12.0 (C-2 ¹ ), 11.0 (C-7 ¹ )

< Example  7> Upper lobe From active fraction  Preparation of Compound of Formula (2)

The active fraction obtained in Example 5 was separated by Florisil column chromatography ( n -hexane: ethyl acetate = 30: 1 to 1: 1 as mobile phase, flow rate 3 ml / min), divided into nine fractions, Silica gel column chromatography ( n -hexane: ethyl acetate = 30: 1 to 1: 1) was performed on the 3-5 fraction to separate 22 mg of Pheophorbide A ethyl ester of Chemical Formula 2.

¹ H-NMR (300 MHz, Chloroform-d) δ 9.42 (1H, s, H-10), 9.26 (1H, s, H-5), 8.54 (1H, s, H-20), 7.91 (3H, dd, J = 17.8, 11.6Hz, H-3 ¹ ), 6.26 (1H, s, H-13 ² ), 6.15 (1H, d, J = 17.8Hz, H-3 ² _a ), 6.11 (1H, d , J = 11.6 Hz, H-3 ² _b ), 4.46 (1H, q, J = 7.2 Hz, H-18), 4.13 (1H, br.d, J = 8.1 Hz, H-17), 4.02 (2H , q, J = 6.9 Hz, H-17 ⁴ ), 3.89 (3H, s, H-13 ⁴ ), 3.66 (3H, s, H-12 ¹ ), 3.54 (2H, q, J = 7.4Hz, H -8 ¹ ), 3.36 (3H, s, H-2 ¹ ), 3.13 (3H, s, H-7 ¹ ), 2.61 (1H, m, H-17 ¹ _a ), 2.43 (1H, m, H- 17 ² _a ), 2.33 (1H, m, H-7 ¹ _b ), 2.24 (1H, m, 17 ² _b ), 1.82 (3H, d, J = 7.2 Hz, H-18 ¹ ), 1.64 (3H, t, J = 6.9 Hz, H-17 ⁵ ), 1.11 (3H, t, J = 7.4 Hz, H-8 ² ), -1.68 (2H, br.s, NH)

¹³ C-NMR (125 MHz, Chloroform-d) δ 189.9 (C-13 ¹ ), 173.2 (C-17 ³ ), 172.4 (C-19), 169.8 (C-13 ³ ), 161.4 (C-16) , 155.8 (C-6), 151.1 (C-14), 149.9 (C-9), 145.3 (C-8), 142.2 (C-1), 138.1 (C-11), 136.7 (C-4), 136.4 (C-7), 136.3 (C-3), 132.0 (C-2), 129.2 (C-3 ¹ ), 129.2 (C-12), 129.1 (C-13), 122.9 (C-3 ² ) , 105.4 (C-15), 104.6 (C-10), 97.7 (C-5), 93.3 (C-20), 64.9 (C-13 ² ), 60.7 (C-17 ⁴ ), 53.1 (C-13 ⁴ ), 51.3 (C-17), 50.3 (C-18), 31.4 (C-17 ¹ ), 30.0 (C-17 ² ), 23.3 (C-18 ¹ ), 19.6 (C-8 ¹ ), 17.6 (C-8 ² ), 14.3 (C-17 ⁵ ), 12.3 (C-12 ¹ ), 12.3 (C-2 ¹ ), 11.4 (C-7 ¹ )

< Example  8> Upper lobe From active fraction  Preparation of Compound of Formula 3

The active fraction obtained in Example 5 was separated by Florisil column chromatography ( n -hexane: ethyl acetate = 30: 1 to 1: 1 as mobile phase, flow rate 3 ml / min), divided into nine fractions, Silica gel column chromatography ( n -hexane: ethyl acetate = 5: 1 to 1: 1) was performed on the 3-6 fractions to separate 130 mg of hydroxy-phenophorbide A ethyl ester of Chemical Formula 3.

¹ H-NMR (300 MHz, Chloroform-d) δ 9.51 (1H, s, H-10), 9.34 (1H, s, H-5), 8.63 (1H, s, H-20), 7.92 (3H, dd, J = 17.8, 11.6 Hz, H-3 ¹ ), 6.23 (1H, d, J = 17.8 Hz, H-3 ² _a ) 6.13 (1H, d, J = 11.6 Hz, H-3 ² _b ), 5.54 (0.6H, s, H-13 ² ), 5.34 (0.4H, s, H-13 ² ), 4.69 (0.4H, m, H-17), 4.48 (1H, q, J = 7.0 Hz, H -18), 4.01 (2H, q, J = 7.0 Hz, H-17 ⁴ ), 3.71 (3H, s, H-13 ⁴ ), 3.68 (0.6H, m, H-17), 3.62 (3H, s , H-12 ¹ ), 3.57 (2H, q, J = 7.6 Hz, H-8 ¹ ), 3.39 (3H, s, H-2 ¹ ), 3.13 (3H, s, H-7 ¹ ), 2.89 ( 1H, m, H-17 ¹ _a ), 2.57 (1H, m, H-17 ² _a ), 2.42 (1H, m, H-7 ¹ _b ), 2.01 (1H, m, 17 ² _b ), 1.61 ( 3H, d, J = 7.0 Hz, H-18 ¹ ), 1.61 (3H, t, J = 7.6 Hz, H-8 ² ), 1.17 (3H, t, J = 7.0 Hz, H-17 ⁵ ),- 1.88 (br.s, NH)

¹³ C-NMR (125 MHz, Chloroform-d) δ 192.0 (C-13 ¹ ), 189.9 (C-17 ³ ), 172.8 (C-19), 172.4 (C-13 ³ ), 162.4 (C-16) , 155.5 (C-6), 151.0 (C-14), 150.2 (C-9), 145.1 (C-8), 142.1 (C-1), 137.8 (C-11), 136.4 (C-7), 136.3 (C-3), 136.2 (C-4), 131.9 (C-2), 129.6 (C-3 ¹ ), 129.3 (C-12), 129.3 (C-13), 122.8 (C-3 ² ) , 107.6 (C-15), 104.2 (C-10), 97.9 (C-5), 93.6 (C-20), 60.6 (C-17 ⁴ ), 60.4 (C-13 ² ), 53.8 (C-13 ⁴ ), 51.8 (C-17), 50.3 (C-18), 31.6 (C-17 ¹ ), 29.7 (C-17 ² ), 22.7 (C-18 ¹ ), 19.4 (C-8 ¹ ), 17.4 (C-8 ² ), 14.1 (C-17 ⁵ ), 12.3 (C-12 ¹ ), 12.1 (C-2 ¹ ), 11.2 (C-7 ¹ )

< Example  9> Upper lobe From active fraction  Preparation of Compound of Formula 4

The active fraction obtained in Example 5 was separated by Florisil column chromatography ( n -hexane: ethyl acetate = 30: 1 to 1: 1 as mobile phase, flow rate 3 ml / min), divided into nine fractions, Silica gel column chromatography ( n -hexane: ethyl acetate = 30: 1 ~ 1: 1) was performed on the 3-3 fractions to separate 4 mg of didehydro- rodochlorine ethylmethyl ester of the formula (4).

¹ H-NMR (300 MHz, Chloroform-d) δ 9.86 (1H, s, H-15), 9.72 (1H, s, H-10), 9.59 (1H, s, H-5), 8.76 (1H, s, H-20), 8.05 (3H, dd, J = 18.0, 12.0 Hz, H-3 ¹ ), 6.33 (1H, d, J = 18.0 Hz, H-3 ² _a ) 6.14 (1H, d, J = 12.0 Hz, H-3 ² _b ), 4.54 (1H, q, J = 7.2 Hz, H-18), 4.39 (3H, s, H-13 ² ), 4.14 (2H, q, J = 7.2 Hz, H-17 ⁴ ), 4.13 (1H, br.d, J = 8.1 Hz, H-17), 3.75 (2H, q, J = 7.8 Hz, H-8 ¹ ), 3.66 (3H, s, H-12 ¹ ), 3.36 (3H, s, H-2 ¹ ), 3.13 (3H, s, H-7 ¹ ), 2.79 (1H, m, H-17 ² _a ), 2.63 (1H, m, H-17 ¹ _a ), 2.52 (2H, m, H-17 ² _b ), 2.52 (2H, m, H-17 ¹ _b ), 1.93 (3H, d, J = 7.2 Hz, H-18 ¹ ), 1.73 (3H, t, J = 7.8 Hz, H-8 ² ), 1.22 (3H, t, J = 7.2 Hz, H-17 ⁵ ), 0.14 (1H, br.s, NH)

¹³ C-NMR (125 MHz, chloroform-d) δ 173.7 (C-17 ³ ), 171.5 (C-19), 167.3 (C-13 ¹ ), 166.9 (C-14), 154.4 (C-6), 149.7 (C-9), 145.0 (C-8), 140.2 (C-1), 140.1 (C-11), 137.2 (C-16), 136.3 (C-7), 135.1 (C-3), 134.7 (C-4), 130.5 (C-12), 130.3 (C-2), 129.6 (C-3 ¹ ), 122.0 (C-3 ² ), 119.1 (C-13), 102.2 (C-10), 99.0 (C-5), 96.4 (C-15), 93.1 (C-20), 60.6 (C-17 ⁴ ), 55.0 (C-17), 52.1 (C-13 ² ), 48.8 (C-18) , 32.7 (C-17 ² ), 31.5 (C-17 ¹ ), 23.7 (C-18 ¹ ), 19.8 (C-8 ¹ ), 17.9 (C-8 ² ), 14.4 (C-17 ⁵ ), 13.8 (C-12 ¹ ), 12.3 (C-2 ¹ ), 11.5 (C-7 ¹ )

< Example  10> Upper lobe From active fraction  Preparation of Compound of Formula 5

The active fraction obtained in Example 5 was separated by Florisil column chromatography ( n -hexane: ethyl acetate = 30: 1 to 1: 1 as a mobile phase, flow rate 3 ml / min), divided into nine fractions, Silica gel column chromatography ( n -hexane: ethyl acetate = 30: 1 to 1: 1) was performed on the 3-3 fraction to separate 140 mg of hydroxy perobide A methyl ester of Chemical Formula 5.

¹ H-NMR (300 MHz, Chloroform-d) δ 9.47 (1H, s, H-10), 9.29 (1H, s, H-5), 8.61 (1H, s, H-20), 7.89 (1H, dd, J = 18.0 Hz, 12.0 Hz, H-3 ¹ ), 6.21 (1H, d, J = 18.0 Hz, H-3 ² ), 6.11 (1H, d, J = 12.0 Hz, H-3 ² ), 5.50 (1H, s, H-13 ^{2 /} OH), 4.48 (1H, q, J = 7.2 Hz, H-18), 4.15 (1H, dd, J = 9.0, 2.0 Hz, H-17), 3.70 ( 3H, s, H-12 ¹ ), 3.65 (3H, s, H-13 ⁴ ), 3.62 (3H, s, H-17 ⁴ ), 3.40 (2H, q, J = 7.7 Hz, H-8 ¹ ) , 3.37 (3H, s, H-2 ¹ ), 3.10 (3H, s, H-7 ¹ ), 2.96 (1H, m, H-17 ¹ a), 2.57 (1H, m, H-17 ² a) , 2.34 (1H, m, H-17 ¹ b), 2.27 (1H, m, H-17 ² b), 1.59 (3H, d, J = 7.2 Hz, H-18 ¹ ), 1.59 (2H, t, J = 7.7 Hz, H-8 ² ), -1.91 (2H, br.s, NH)

¹³ C-NMR (125 MHz, Chloroform-d) δ 192.2 (C-13 ¹ ), 174.2 (C-17 ³ ), 173.0 (C-1), 172.6 (C-13 ³ ), 162.6 (C-19) , 155.5 (C-9), 151.2 (C-11), 150.0 (C-16), 145.3 (C-8), 142.2 (C-4), 138.0 (C-14), 136.6 (C-3), 136.4 (C-6), 136.4 (C-7), 131.9 (C-2), 129.5 (C-12), 129.2 (C-3 ¹ ), 127.1 (C-13), 123.0 (C-3 ² ) , 107.9 (C-15), 104.4 (C-10), 98.1 (C-5), 93.8 (C-20), 89.2 (C-13 ² ), 53.6 (C-13 ⁴ ), 52.0 (C-17 ), 51.9 (C-17 ⁴ ), 50.5 (C-18), 31.7 (C-17 ¹ ), 31.4 (C-17 ² ), 22.9 (C-18 ¹ ), 19.6 (C-8 ¹ ), 17.6 (C-8 ² ), 12.5 (C-12 ¹ ), 12.3 (C-2 ¹ ), 11.3 (C-7 ¹ )

< Example  11> Upper lobe From active fraction  Preparation of Compound of Formula 6

The active fraction obtained in Example 5 was separated by Florisil column chromatography ( n -hexane: ethyl acetate = 30: 1 to 1: 1 as mobile phase, flow rate 3 ml / min), divided into nine fractions, Silica gel column chromatography ( n -hexane: ethyl acetate = 30: 1 to 1: 1) was performed on the 3-4 fractions to separate 31 mg of the pheophorbide A methyl ester of the formula (6).

¹ H-NMR (300 MHz, Chloroform-d) δ 9.51 (1H, s, H-10), 9.37 (1H, s, H-5), 8.56 (1H, s, H-20), 7.97 (3H, dd, J = 17.8, 11.6Hz, H-3 ¹ ), 6.28 (1H, d, J = 17.8Hz, H-3 ² _a ), 6.25 (1H, s, H-13 ² ), 6.13 (1H, d , J = 11.6 Hz, H-3 ² _b ), 4.45 (1H, q, J = 7.2 Hz, H-18), 4.14 (1H, br.d, J = 8.1 Hz, H-17), 3.87 (32H , s, H-17 ⁴ ), 3.71 (3H, s, H-13 ⁴ ), 3.59 (3H, s, H-12 ¹ ), 3.53 (2H, q, J = 7.4 Hz, H-8 ¹ ), 3.39 (3H, s, H-2 ¹ ), 3.20 (3H, s, H-7 ¹ ), 2.61 (1H, m, H-17 ¹ _a ), 2.55 (1H, m, H-17 ² _a ), 2.35 (1H, m, H-7 ¹ _b ), 2.21 (1H, m, 17 ² _b ), 1.80 (3H, d, J = 7.2 Hz, H-18 ¹ ), 1.26 (3H, t, J = 7.4 Hz, H-8 ² ), -1.68 (2H, br.s, NH)

¹³ C-NMR (125 MHz, Chloroform-d) δ 189.8 (C-13 ¹ ), 173.5 (C-17 ³ ), 172.4 (C-19), 169.8 (C-13 ³ ), 161.4 (C-16) , 155.8 (C-6), 151.1 (C-14), 149.8 (C-9), 145.4 (C-8), 142.2 (C-1), 138.1 (C-11), 136.7 (C-4), 136.4 (C-7), 136.3 (C-3), 132.0 (C-2), 129.2 (C-3 ¹ ), 129.2 (C-12), 129.1 (C-13), 123.0 (C-3 ² ) , 105.3 (C-15), 104.6 (C-10), 97.7 (C-5), 93.3 (C-20), 64.9 (C-13 ² ), 53.1 (C-13 ⁴ ), 51.9 (C-17 ⁴ ), 51.3 (C-17), 50.3 (C-18), 31.2 (C-17 ¹ ), 30.0 (C-17 ² ), 23.3 (C-18 ¹ ), 19.6 (C-8 ¹ ), 17.6 (C-8 ² ), 12.3 (C-12 ¹ ), 12.3 (C-2 ¹ ), 11.4 (C-7 ¹ )

< Experimental Example  1> MCH -1 receptor binding inhibitory effect

In order to determine the MCH-1 receptor binding inhibitory activity of the leaf extract, fractions or active substances thereof according to the present invention was carried out as follows.

The buffer solution is prepared by two types of washing solution (25 mM HEPES pH 7.4, 5 mM MgCl ₂ , 1 mM CaCl ₂ ) and experimental solution (adding 0.5% BSA to the washing solution), and MCH-1 receptor (Euroscreen, Gosselies). , Belgium) and 1 μM europium labeled melanin enriched hormone (Europium (Eu) -labeled MCH, PerkinElmer, Turku, Finland) and 1 mM melanin enriched hormone (MCH, # 070-47, Phoenix, Belmont CA, USA) Was kept at 4 ° C. 1 μM Eu-labeled MCH and 1 mM MCH were diluted to 8 nM (final reaction concentration 2 nM) and 2 μM (final reaction concentration 0.5 μM), respectively. The buffer used in all dilutions and preparations was the experimental solution, and the wash solution was used only when the plate was washed last.

MCH-1 receptor (200 assays / vial) was first homogenized by diluting in 1 ml of experimental solution. Thereafter, the leaf extract prepared in Example 1, 4-11 or a compound separated therefrom was prepared as a test substance, and then the filter plate attached microplate (Multiwell 96 well filter plates PN5020, Pall Co. Ann Arbor MI, USA). Using an 8-channel pipette (multi 8-channel, Eppendorf, Hamburg, Germany), the extract of the upper leaf prepared in Examples 1, 4-11 or separated therefrom so that the total volume is 100 μl per well Compounds were dispensed. In this case, non-specific binding is 25 μl of Eu-labeled MCH and 50 μl of receptor and 25 μl of MCH, and total binding is 25 μl of 10% DMSO experimental solution and 25 μl of Eu-labeled MCH and 50 μl of receptor was mixed. Finally, the experimental group mixed 25 μl of test substance with 25 μl of Eu-labeled MCH and 50 μl of receptor. After all of this was shaken gently for 15 seconds and reacted at room temperature for 90 minutes.

At the end of the reaction, the plate was partially cleaned by applying pressure to a self-made washer (microplate washer, EMBLA, Molecular Devices). The wash solution was filtered three times at 300 μl per well. This process removes the remaining Eu-labeled MCH without reacting. Then, the bottom of the water was wiped off and the dissociation solution (DELFIA Enhancement solution, PerkinElmer, Turku, Finland) was added to 150 μl per well. After shaking for 10 minutes or measured, or reacted for 2 to 4 hours as it is at room temperature to measure the time-resolved fluorescence (TRF) value. As a measurement method, the emission wavelength was 615 nm and the excitation wavelength was 340 nm, which was measured by a multi-label fluorometer (multilabel counter, Victor2, PerkinElmer, Turku, Finland). Differential fluorescence inhibition was calculated by the following equation (1).

% Differential Fluorescence Inhibition = [(Total Binding Average Value-Test Material's Differential Fluorescence Value) / (Total Binding Average Value-Nonspecific Binding Average Value)] × 100

After primarily measuring the differential fluorescence inhibition at 10 μM, the IC ₅₀ value was calculated only for test substances inhibited by 50% or more. For the test substances inhibited by more than 50%, the differential fluorescence values were obtained at four concentrations of 0.01, 0.1, 1.0, and 10.0 μM, respectively, and IC ₅₀ values were calculated through linear regression analysis. The results are shown in Tables 1 and 2. .

sample IC ₅₀ (μg / ml) Ethanol Extract of Upper Leaf 2.28 ± 1.04 Ethanol Extract of Upper Leaf → Dichloromethane Fraction 1.57 ± 0.77 Ethanol Extract of Upper Leaf → Dichloromethane Fraction → Hexane Fraction 17.12 ± 9.89 Ethanol Extract of Upper Leaf → Dichloromethane Fraction → Methanol Fraction 0.97 ± 0.46 Ethanol Extract of Upper Leaf → Ethyl Acetate Fraction 6.67 ± 1.79 Ethanol Extract of Upper Leaf → Butanol Fraction > 30 Ethanol Extract of Upper Leaf → Water Fraction > 30 Methanol Extract of Upper Leaf 3.98 ± 1.02 Methanol Extract of Upper Leaf → Dichloromethane Fraction 2.42 ± 0.89 Methanol Extract of Upper Leaf → Ethyl Acetate Fraction 6.34 ± 2.33 Methanol Extract of Upper Leaf → Butanol Fraction > 30 Methanol Extract of Upper Leaf → Water Fraction > 30

sample IC ₅₀ (μM) Perpurin-7 dimethyl ethyl ester 0.26 ± 0.11 Pheophorbide A ethyl ester 0.24 ± 0.08 Hydroxy-Peophorbide A Ethyl Ester 0.11 ± 0.02 Pheophorbide A Methyl Ester 4.03 ± 2.48 Hydroxy-Peophorbide A Methyl Ester 0.33 ± 0.29 Didehydro-Rodochlorine Ethyl Methyl Ester 14.94 ± 4.24

As shown in Table 1, the IC ₅₀ value for the MCH-1 receptor binding inhibitory activity of the upper leaf ethanol extract, and the dichloromethane fraction and ethyl acetate fraction thereof according to the present invention is excellent MCH by showing 0.97 ~ 6.67 ㎍ / ml It can be seen that -1 receptor binding inhibitory action. Butanol fraction and water fraction showed little MCH-1 receptor binding inhibitory activity (IC ₅₀ > 30 ㎍ / ml). The IC ₅₀ values of MCH-1 receptor binding inhibitory activity of the methanol extract of the upper leaf, and its dichloromethane fractions and ethyl acetate fractions ranged from 2.42 to 6.34 ㎍ / ml, but the butanol fraction and water fraction inhibited MCH-1 receptor binding. It showed little action (IC ₅₀ > 30 ㎍ / ml), showing a similar pattern to that of the ethanol extract of the lobe.

As shown in Table 2, IC ₅₀ on the MCH-1 receptor binding inhibitory activity of the compounds isolated from the extract of the upper leaf according to the present invention Values of 0.11 to 14.94 µg / ml indicate that the MCH-1 receptor binding inhibitory activity is excellent. In particular, compounds such as Perpurin-7 dimethyl ethyl ester, pheophorbide A ethyl ester, hydroxy-phenophorbide A ethyl ester, and hydroxy- pheophorbide A methyl ester are about 10 times stronger than the extract effect. appear.

Therefore, the upper leaf extract according to the present invention or a compound isolated therefrom exhibits excellent MCH-1 receptor binding inhibitory action, and thus can be usefully used for suppressing appetite, preventing and improving depression, or preventing and improving anxiety.

< Experimental Example  2> Measuring appetite suppression effect and weight loss effect

In order to determine the appetite suppression effect and weight loss effect of the upper leaf extract, fractions or active substances thereof according to the present invention was carried out as follows.

Dietary-induced obese mice were fed a high-fat diet (60% fat in total calories: D12492. Research diet) for 9 weeks from 5 to 14 weeks of age, and within a short period of time, the control group (6.5% fat in total calories: Purina). A model of obesity similar to severe obesity in humans was completed, with significant weight gain (more than 25%) compared to Rodent chow 5008 (Ralston-purina). The mice used were Jacson lab, USA. C57BL / 6J mice were purchased at 4 weeks of age and were used from 5 weeks of age after one week of adaptation. The mean weight of 5 weeks old before the experiment was 18.7 ± 0.88 g, and the mean weight of 14 weeks old was 37.0 ± 0.2 g (64), which was significantly different from the average body weight of 29.6 ± 0.5 g (9). Seemed. The number of dietary-induced obese mice in each group was 10-12. The diet was maintained as a high fat diet (60% fat-containing diet of total calories: D12492. Research diet), and the extract of the upper leaf prepared in Example 1 was dissolved in 0.5% tween 80 twice a day 100, 250 , 500 mg / kg orally, and feeding and weight changes were measured at 32-day 2-day intervals. The temperature of the feeding room was maintained at 25 ± 2 ℃ and the day / night cycle was crossed every 12 hours, water and high-fat diet was free to eat. The control group was orally administered with only 0.5% tween 80 solution, and the control group was treated with oral sibutramin 3 mg / kg. Weight change and feeding amount of mice in each group were measured daily for 32 days.

The measurement results are shown in FIGS . 1 and 2 .

Figure 1 is a graph showing the weight change of the mouse for administration of the upper leaf extract according to the present invention. As shown in Figure 1 , the upper leaf extract 100 mg / kg administration group according to the present invention did not show a significant difference compared to the control group, the weight of the obese mice administered 250, 500 mg / kg compared to the control group 2 From day onwards, it gradually decreased. In particular, the 250 mg / kg administration group showed a significant difference from 4 days of administration, 500 mg / kg administration group from 14 days. On the other hand, sibutramine of the comparison group showed a significant decrease after 2 days of administration, after which weight loss began to slow down, and from day 26, there was no significant difference compared to the control group.

Figure 2 is a graph showing the change in feeding amount of the mouse to the administration of the extract of the upper leaf to the present invention. As shown in Figure 2 , the feed intake was reduced in all the administration groups administered 100, 250, 500 mg / kg of the upper leaf extract according to the present invention, it was observed to show a sustained appetite decrease throughout the experimental period. On the other hand, the sibutramine-administered group showed a sharp decrease in feed intake, and then increased rapidly.

Therefore, the upper leaf extract according to the present invention can be usefully used for preventing and improving obesity because it induces weight loss and decreased appetite.

Examples of formulations for the composition of the present invention are illustrated below.

< Formulation example  1> Manufacture of food

<1-1> Wire

Brown rice, barley, glutinous rice, and yulmu were alphad by a known method, and then dried and roasted to make a powder having a particle size of 60 mesh. Black beans, black sesame seeds, and perilla were also steamed and dried in a known manner, and then ground to a powder having a particle size of 60 mesh.

The upper leaf extract of the present invention or the compound of Chemical Formulas 1 to 6 were concentrated under reduced pressure in a vacuum concentrator, and the dried product obtained by drying with a spray hot air dryer was pulverized to a particle size of 60 mesh with a grinder to obtain a dry leaf extract powder.

Granules were prepared by combining the grains, seeds and leaf extracts prepared above or the dry powder of the compound of Formulas 1 to 6 in the following ratios.

Cereals: Brown rice 30% by weight, barley 15% by weight, barley 20% by weight, glutinous rice 9% by weight,

Seeds: perilla 7% by weight, black beans 8% by weight, black sesame 7% by weight,

3% by weight of the dry leaf extract or the dry powder of the compound of formulas 1-6, Ganoderma lucidum 0.5%

<1-2> Chewing gum

Chewing gum was prepared in a conventional manner by combining 20% by weight of gum base, 76.9% by weight of sugar, 1% by weight of perfume, and 2% by weight of water, and 0.1% by weight of the leaf extract of the present invention.

<1-3> candy

Candy was prepared in a conventional manner by combining 60% by weight of sugar, 39.8% by weight of starch syrup, and 0.1% by weight of fragrance, and 0.1% by weight of the upper leaf extract of the present invention or the compound of Formulas 1-6.

<1-4> Biscuits

Force 25.59% by weight, 22.22% by weight of gravity, 4.80% by weight, white salt 0.73% by weight, 0.78% by weight, palm shortening 11.78%, ammonium 1.54% by weight, 0.17% by weight sodium bisulfite 0.16% by weight , Rice flour 1.45%, Vitamin B₁0.0001%, Vitamin B₂0.0001%, Milk flavor 0.04%, Water 20.6998%, Whole milk powder 1.16%, Substitute milk powder 0.29%, Monobasic calcium phosphate 0.03% , Biscuits were prepared by combining 0.29 wt% of spray salt and 7.27 wt% of spray oil with 1 wt% of the upper leaf extract of the present invention or the compound of Formulas 1-6.

<1-5> health drink

0.26% by weight of honey, 0.0002% by weight of thioctoamide, 0.0004% by weight of nicotinic acid, 0.0001% by weight of sodium riboflavinate, 0.0001% by weight of pyridoxine hydrochloride, 0.001% by weight of inositol, 0.002% by weight of orthoic acid and 98.7362% by weight of water 1% by weight of the extract of the upper leaves or the compound of Formulas 1 to 6 were prepared in a conventional manner for the health beverage.

<1-6> Health Supplement

55% by weight of spirulina, 10% by weight of guar gum enzyme digestion, 0.01% by weight of vitamin B ₁ hydrochloride, 0.01% by weight of vitamin B ₆ hydrochloride, 0.23% by weight of DL-methionine, 0.7% by weight of magnesium stearate, 22.2% by weight of lactose and 1.85% by weight of corn starch % And 10% by weight of the leaf extract of the present invention or the compound of Formulas 1 to 6 were prepared in a conventional dietary supplement.

1 is a graph showing the weight loss of dietary-induced obese mice according to the dosage of the upper leaf extract according to an embodiment of the present invention.

Figure 2 is a graph showing the change in feeding amount of dietary-induced obese rats according to the dosage of the extract of the upper lobe according to an embodiment of the present invention.

Claims (15)

delete delete delete The upper leaf is extracted with methanol or ethanol as a solvent, and then methanol or ethanol is blown off and the distilled water is suspended in the extract residue, which is then selected from the group consisting of ethyl acetate, dichloromethane and butanol. Fractions obtained by fractionation as an active ingredient, comprising a compound of any one of formulas 1 to 6, and antagonistic activity against melanin Concentration Hormone receptor subtype-1 (MCH-1) Food composition for suppressing appetite, preventing and improving depression, or preventing and improving anxiety, characterized in that: [Formula 1]

[Formula 2]

(3)

[Formula 4]

[Chemical Formula 5]

[Formula 6]

. The food composition of claim 4, wherein the organic solvent is dichloromethane. The dichloromethane fraction obtained by diluting the upper leaf extract in distilled water and distilling with dichloromethane is the concentration gradient of the mixed solvent of n-hexane and ethyl acetate. Food composition, characterized in that the active fraction obtained by performing silica gel column chromatography. A food composition for suppressing appetite, preventing and improving depression, or preventing and improving anxiety, containing any one of the following Formulas 1 to 6 or a mixture thereof as an active ingredient: [Formula 1]

[Formula 2]

(3)

[Formula 4]

[Chemical Formula 5]

[Formula 6]

. The method of claim 7, wherein the compound of Formula 1 to 6 Adding methanol or ethanol to the upper leaf to obtain an upper leaf extract (step 1); Suspending the upper leaf extract obtained in step 1 in water and fractionating with dichloromethane to obtain a dichloromethane fraction (step 2); Dichloromethane fractions obtained in step 2 using a mixed solvent of n- hexane and ethyl acetate as a mobile phase to perform a concentration gradient column chromatography to obtain an active fraction (step 3); Separating the active fraction from the active fraction obtained in step 3 by performing concentration gradient column chromatography again using a mixed solvent of n-hexane and ethyl acetate as a mobile phase (step 4); And The fraction separated in step 4 is a mixed solvent of n-hexane and ethyl acetate as a mobile phase, but performing a silica gel column chromatography with different concentration gradients to separate the compound of formula 1-6 (step 5) Food composition, which is prepared by a method comprising. delete The food composition of claim 8, wherein step 3 is performed by silica gel column chromatography using a concentration gradient of a mixed solvent of n-hexane and ethyl acetate as a mobile phase of 100: 1 to 1: 1. The method of claim 8, wherein step 4 is the concentration gradient of the mixed solvent of n-hexane and ethyl acetate in the third fraction of the first to fourth fractions of the active fraction obtained in step 3 to 30: 1 to 1: 1 Food composition, characterized in that separated by Florisil column chromatography. The food composition of claim 8, wherein the concentration gradient of step 5 is 15: 1 to 1: 1, 30: 1 to 1: 1, and 5: 1 to 1: 1. delete The food composition according to claim 4, wherein the upper leaf fraction is a hexane fraction obtained by suspending the upper leaf extract in distilled water and distilling the dichloromethane fraction obtained by distilling with dichloromethane. The food composition according to claim 4, wherein the upper leaf fraction is a methanol fraction obtained by suspending the upper leaf extract in distilled water and distilling the dichloromethane fraction obtained by distilling with dichloromethane.

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