KR20120069964A - Composition for anti-obesity - Google Patents

Abstract

The present invention provides an anti-obesity composition containing at least one selected from 1-methoxyindole-3-acetonitrile or 4-methoxyindole-3-acetonitrile as an active ingredient. The composition of the present invention has the advantage of having an excellent, safe anti-obesity effect.

Description

Composition for anti-obesity {Composition for anti-obesity}

The present invention relates to an anti-obesity composition, and more particularly, an anti-obesity composition containing at least one selected from 1-methoxyindole-3-acetonitrile or 4-methoxyindole-3-acetonitrile as an active ingredient. It is about.

Anti-obesity is meant to encompass the prevention, prevention or treatment of obesity, obesity means that the excess calories are consumed compared to the calories consumed by the excess calories are accumulated in the form of fat in the body.

Currently, appetite suppressants, heat generators, and diuretics are used as treatments for obesity, and the most commonly used appetite suppressants may exhibit side effects such as an increase in blood pressure. Therefore, there is an increasing need to use food ingredients, which have long-term safety and physiological function, for anti-obesity use.

On the other hand, the anti-obesity use of 1-methoxyindole-3-acetonitrile or 4-methoxyindole-3-acetonitrile is not known.

The problem to be solved by the present invention is to provide a composition for anti-obesity.

The present invention relates to 1-methoxyindole-3-acetonitrile (1-MIA) or 4-methoxyindole-3-acetonitrile (4-MIA). It provides one or more anti-obesity uses selected from.

The 1-methoxyindole-3-acetonitrile may be represented by Formula I, and 4-methoxyindole-3-acetonitrile may be represented by Formula II.

(I)

≪ RTI ID = 0.0 &

In addition, the present invention provides a composition for anti-obesity containing at least one selected from 1-methoxyindole-3-acetonitrile or 4-methoxyindole-3-acetonitrile as an active ingredient.

The active ingredient may be isolated from a turnip ( Brassica campestris spp rapa), preferably fortified turnip. The fortified turnip means a turnip grown or grown in Ganghwado.

Extracting the 1-methoxyindole-3-acetonitrile with methanol; Partitioning and extracting the methanol extract into ethyl acetate / H ₂ O, and then partitioning and extracting the H ₂ O layer into n-BuOH / H ₂ O; Concentrating the partitioned extracted layer under reduced pressure to obtain an ethyl acetate fraction, n-butanol fraction and water fraction; Silica gel column chromatography on the ethyl acetate fraction (n-hexane-ethyl acetate = 10: 1 → 5: 1 → 3: 1 → 1: 1 → CHCl ₃ -methanol = 10: 1 → 7: 1 → 5: 1 → 3: 1 → 2: 1 → 1: 1) to obtain 18 fractions (BE1 ~ BE18); And the seventh fraction (BE7 fraction) of the 18 fractions may be separated from the turnip by a separation method comprising the step of purification by separating with octadecyl silica gel.

In addition, the 4-methoxyindole-3-acetonitrile is the step of extracting turnip with methanol; Partitioning and extracting the methanol extract into ethyl acetate / H ₂ O, and then partitioning and extracting the H ₂ O layer into n-BuOH / H ₂ O; Concentrating the partitioned extracted layer under reduced pressure to obtain an ethyl acetate fraction, n-butanol fraction and water fraction; Silica gel column chromatography on the ethyl acetate fraction (n-hexane-ethyl acetate = 10: 1 → 5: 1 → 3: 1 → 1: 1 → CHCl ₃ -methanol = 10: 1 → 7: 1 → 5: 1 → 3: 1 → 2: 1 → 1: 1) to obtain 18 fractions (BE1 ~ BE18); And it may be separated from the turnip by a separation method comprising the step of purifying with octadecyl silica gel for the ninth fraction (BE9 fraction) of the 18 fractions.

The anti-obesity is meant to encompass obesity suppression, obesity treatment, improvement or prevention, but is not limited thereto, by at least one selected from anti-adipogenic effect, lipolysis, or exothermic reaction. It can have an anti-obesity effect. The anti-localization effect means an effect of inhibiting fat precursor cells (preadipocyte) is differentiated into mature adipocytes (adipocyte) to accumulate fat.

The composition of the present invention may be a food composition or a pharmaceutical composition, in the food composition, the treatment or prevention is meant to include improvement or prevention. The food composition may be a health functional food, the formulation may be prepared according to a conventional method, and then dried with a carrier and encapsulated, or may be formulated in the form of other tablets, granules, powders, beverages, porridge, etc. Besides, it can be manufactured in any food form.

In addition, the present invention is 1-methoxyindole-3-acetonitrile (1-methoxyindole-3-acetonitrile; 1-MIA) or 4-methoxyindole-3-acetonitrile (4-methoxyindole-3-acetonitrile; 4- MIA) provides the use for the production of one or more anti-obesity agents selected from.

In addition, the present invention provides a pharmaceutically effective amount of 1-methoxyindole-3-acetonitrile (1-MIA) or 4-methoxyindole-3-acetonitrile (4-methoxyindole- Provided is an anti-obesity method comprising the step of administering to a mammal (including human) in need of at least one selected from 3-acetonitrile (4-MIA).

Unless otherwise stated, the content of the composition of the present invention is equally applicable to the use of the present invention, the anti-obesity agent preparation, and the anti-obesity method.

The composition may be administered orally or parenterally to mammals including humans, and the active ingredient may be formulated in combination with a pharmaceutically acceptable carrier. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents and surfactants which are commonly used may be used. Solid form preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid form preparations include at least one excipient such as starch, calcium carbonate, sucrose, lactose, and It is prepared by adding gelatin and the like. Lubricants such as magnesium and talc are also used. Liquid preparations for oral administration include suspensions, solvents, emulsions and syrups, and may include various excipients such as wetting agents, sweeteners, fragrances and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. have. Formulations for parenteral administration include injectable solutions, suspensions, emulsions, lyophilizers, nasal washes and suppositories. Injectable solutions, suspensions and emulsions can be prepared by mixing water, non-aqueous solvents or suspending solvents with the active ingredient. As non-aqueous solvents and suspending solvents, vegetable oils such as propylene glycol, polyethylene glycol and olive oil, and ethyl oleate Injectable esters such as and the like. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerol, gelatin and the like can be used. The composition of the present invention may be administered by subcutaneous injection, intravenous injection or intramuscular injection during parenteral administration.

In the compositions, uses and methods of the present invention, it can be used once or divided into several doses of 0.1 to 100 mg / kg of adult per day on an active ingredient basis.

The active ingredient in the composition of the present invention may be contained in 0.01 to 5% by weight.

The active ingredient may be formulated by adding a pharmaceutically or food-acceptable carrier, excipient or diluent, etc., and the formulation may be found in Remington's Pharmaceutical Science (Recent Edition), Mack Publishing Company, Easton PA et al. Reference may be made.

The composition of the present invention has a good anti-obesity effect.

1 is a cell staining result photograph for confirming the anti-adipogenic effect of 1-MIA or 4-MIA.
Figure 2 is a graph showing the absorbance measurement results for confirming the anti-adipogenic effect of 1-MIA or 4-MIA.
Figure 3 is a graph showing the results of RT-PCR confirming the effect of 1-MIA or 4-MIA on β3-AR expression.
Figure 4 is a graph showing the results of RT-PCR confirming the effect of 1-MIA or 4-MIA on HSL expression.
5 is a graph showing the results of RT-PCR confirming the effect of 1-MIA or 4-MIA on ATGL expression.
Figure 6 is a graph showing the results of RT-PCR confirming the effect of 1-MIA on AdipoQ expression.
Figure 7 shows the Western blot results confirming the effect of 1-MIA or 4-MIA on pHSL, HSL, pPKA, PKA expression.

Hereinafter, the present invention will be described in more detail with reference to Examples and Experimental Examples, but the following Examples and Experimental Examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

Silica gel (SiO ₂ ) for column chromatography used in the following examples was Kiesel gel 60 (Merck, Darmstadt, Germany), and octadecyl silica (ODC) gel was LiChroprep RP-18 (Merck, Darmstadt, Germany). Thin layer chromatography (TLC) was performed using Kieselgel 60 F254 and RP-18 F _254s , and all reagents were used as a special reagent. In addition, NMR spectra were measured with a Varian Inova AS 400 (Varian, California, USA), and IR spectra were measured with a Perkin model 599B (Perkin-Elmer, Massachusetts, USA). Melting points were measured by Fisher-John's Melting Point Apparatus (Fisher Scientific, Miami, USA) and EI / MS by JMS-700 (JEOL, Tokyo, Japan). UV lamps were Spectroline (Model ENF-240 C / F, Spectronics Corporation, New York, USA).

In the Examples and Experimental Examples, a T-test was performed on the experimental results to determine that there was statistical significance when the significance difference was less than 5% (p <0.05) or less than 1% (p <0.01).

Example 1 Confirmation of Anti-Obesity Effect by Anti-adipogenic Effect

1-1. Preparation of 1-methoxyindole-3-acetonitrile (abbreviated 1-MIA) or 4-methoxyindole-3-acetonitrile (abbreviated 4-MIA)

100 kg of turnip roots (Ganghwa-gun Agricultural Technology Center) were separated from the shell and the genus, and the genus (48 kg) was chopped and extracted twice with 80% aqueous methanol solution and filtered through a filter paper.

The filtrate was concentrated under reduced pressure at 45 degrees Celsius to obtain a methanol extract. The obtained methanol extract was partitioned and extracted with ethanol acetate (3LX2) / H ₂ O (3L), and the H ₂ O layer was partitioned and extracted with n-BuOH (3LX2) / H ₂ O (3.2L). Each layer was concentrated under reduced pressure to obtain an ethyl acetate fraction (15 g), n-butanol fraction (48 g) and a water fraction.

Silica gel column chromatography (cc) for the ethyl acetate fraction (φ 5 × 22 cm, n-hexane-ethyl acetate = 10: 1 → 5: 1 → 3: 1 → 1: 1 → CHCl ₃ -methanol = 10: 1 → 7: 1 → 5: 1 → 3: 1 → 2: 1 → 1: 1) to obtain 18 fractions (BE1 to BE18). The seventh fraction (BE7 fraction, Ve / Vt 0.36-0.41 (elution volume / total volume)} was purified by ODS cc (φ 5 × 22 cm, methanol-H ₂ O = 1: 1) to obtain BE7-1 {1- MIA, 38 mg, Ve / Vt 0.01-0.12 (elution volume / total volume); TLC (RP-18 F _254s ) Rf-0.75, methanol-water = 5: 1} was separated. In addition, the 9th fraction (BE9 fraction, Ve / Vt 0.47-0.52) was purified by ODS cc (φ 3.5 × 12 cm, methanol-H ₂ O = 1: 1) to obtain BE9-3 (4-MIA, 21 mg, Ve / Vt 0.5-0.63, TLC (RP-18 F _{254 s} ) Rf-0.45, methanol-water = 3: 1) was isolated.

1-MIA; Colorless powder (CHCl ₃ ); mp 149-152 ° C .; EI / MS m / z: 186 [M] ⁺ , 171, 155, 128, 101; IR (CHCl ₃ , cm ⁻¹ ) 2937, 2249, 1454; ¹ H-NMR (400 MHz, CDCl ₃ , δ _H ): 7.47 (1H, d, J = 7.8 Hz, H-7), 7.37 (1H, dd, J = 7.8, 7.8 Hz, H-6), 7.21 ( 1H, dd, J = 7.8, 7.8Hz, H-5), 7.08 (1H, d, J = 7.8Hz, H-4), 7.18 (1H s, H-2), 3.98 (3H, s, H- 12), 3.73 (2H, s, H-10); ¹³ C-NMR (100 MHz, CDCl ₃ , δ _c ): 132.30 (C-8), 123.10 (C-7), 122.32 (C-9), 121.67 (C-6), 120.34 (C-5), 118.24 (C-4), 117.85 (C-11), 108.61 (C-2), 100.27 (C-3), 66.03 (C-12), 14.25 (C-10)

4-MIA; Colorless powder (CHCl ₃ ); mp 141-142 ° C .; EI / MS m / z: 186 [M] ⁺ , 171; IR (CHCl ₃ , cm ⁻¹ ) 3396, 2955, 2850, 1619, 1258; ¹ H-NMR (400 MHz, CDCl ₃ , δ _H ): 7.11 (1H, dd, J = 8.0, 8.0 Hz, H-6), 7.03 (1H, s, H-2), 6.95 (1H, d, J = 8.0 Hz, H-5), 6.49 (1H, d, J = 8.0 Hz, H-7), 4.21 (3H, s, H-12), 4.08 (2H, s, H-10); ¹³ C-NMR (100 MHz, CDCl ₃ , δ _c ): 154.09 (C-4), 137.63 (C-8), 125.12 (C-9), 123.49 (C-7), 121.26 (C-5), 119.18 (C-11), 104.85 (C-3), 104.61 (C-2), 99.67 (C-6), 55.14 (C-12), 16.02 (C-10)

1-2. Experiment confirming the anti-adipogenic effect

The anti-localization effect of 1-MIA and 4-MIA prepared in 1-1. Was confirmed by the following experiment using adipose progenitor cells. 1-MIA and 4-MIA were dissolved in DMSO (dimethyl sulfoxide), respectively.

Preadipocyte 3T3L1 cells (American Type Culture Collection, Rockville, MD, USA) were cultured in DMEM (Gibco) medium containing 10% calf serum (Calbiotech) and 1 mM sodium pyruvate (Gibco). Incubate the cells in a 24 well plate at 100% confluent and incubate for 2 days, and then incubated with Fetal calf serum in DMEM medium with 0.25 μM dexamethasone (DEX, Sigma), 10 μg / ml insulin (Gibco), 0.5 mM 1-methyl-3 isobutylxanthine (IBMX, Sigma) was added to promote differentiation into adipocytes for 3 days. In this 3T3-L1 cell differentiation step, the groups were divided and treated with 1-MIA and 4-MIA prepared in 1-1. At concentrations of 6.3, 12.5, 50.0, and 100.0 μM, respectively. In addition, TNF-alpha was treated at a concentration of 10 nM in a separate group. TNF-alpha was treated with a positive control that inhibited fat accumulation. Thereafter, except for the three stimulants, the culture medium was changed for two to two days at the same medium condition, and then cultured for about 6 to 10 days to accumulate fat. Six days after the start of differentiation, the amount of triglycerides produced in the cells was visually observed through oil red O staining. After the microscopic examination, the oil red O pigment was extracted with 200 μl of isopropanol. rad model 680) was used to measure the absorbance at 520 nm to quantify the degree of fat accumulation. Cells that did not promote differentiation into adipocytes were used as controls.

The results are shown in FIGS. 1 and 2. 1 is a micrograph of the stained cells, Figure 2 is a graph showing the results of absorbance measurements. In FIG. 2, ## is P <0.005 compared to allied cells, * is P <0.05 compared to adipocytes, and ** is P <0.005 compared to adipocytes.

As shown in Figure 1, in the case of the fat cells did not differentiate the fat of the red fat almost did not occur, in the case of fat cells can be seen that a lot of fat accumulated a lot of red. TNF-alpha treatment showed little fat accumulation, with only a blue nucleus. Treatment with 1-MIA and 4-MIA, respectively, tended to reduce fat accumulation in a concentration-dependent manner.

In addition, as shown in Figure 2, it can be seen that the fat accumulation is significantly increased in the case of fat cells, compared to the case of all-cell cells without fat differentiation. TNF-alpha treatment showed cell-level cell levels, and treatment with 1-MIA and 4-MIA showed a decrease in fat accumulation in a concentration-dependent manner.

As a result, it can be seen that 1-MIA and 4-MIA have an anti-obesity effect by reducing fat accumulation of adipocytes. Therefore, it can be seen that the composition of the present invention is effective in anti-obesity.

<Example 2> anti-obesity effect confirmed by exothermic reaction or lipolysis

Changes in the expression levels of beta3-AR associated with lipolysis, heat generation, changes in expression levels of HSL and ATGL, lipolytic enzymes, and AdipoQ, a weight loss-related gene, were measured. To determine the anti-obesity effect by exothermic reaction or lipolysis.

2-1. Preparation of 1-MIA and 4-MIA

It prepared in the same manner as in 1-1.

2-2. Measurement of changes in expression levels of β3-AR, HSL, ATGL, and AdipoQ

β3-AR (β3-adrenergic receptor) is a G-protein-related receptor and is known to be deeply involved in lipolysis and heat generation in adipocytes by adrenergic signals (Giacobino, 1995). Hormone sensitive lipase (AT) and Adipose triglyceride lipase (ATGL) play an important role in the regulation of lipolysis in adipocytes of rodents. ATGL is essential for maintaining basic lipolytic activity in humans, while HSL is most responsive to lipolysis stimulation. Characterized by enzymes (Arnerand Langin, 2007). AdipoQ (adiponectin), a type of adipocytokine, is known to be associated with weight loss and body fat content (Yokohama et al., 2004).

Therefore, in this experiment, we tried to confirm the anti-obesity effect by changing the expression level of β3-AR, HSL, ATGL, AdipoQ during 1-MIA and 4-MIA treatment. 1-MIA and 4-MIA prepared in 2-1. Were dissolved in DMSO (dimethyl sulfoxide), respectively, and the final concentration of DMSO was 0.1% or less during cell experiments. The cells used in the experiments were prepared in the same manner as in Example 1-2, and instead of treating 1-MIA and 4-MIA at concentrations of 6.3, 12.5, 50.0, and 100.0 μM, respectively, when processing samples for the cells. Sample treatment was carried out in the same manner as in Example 1-2, except that the treatment was carried out at the concentrations described in FIGS. 3 to 6.

Changes in gene expression following treatment with 1-MIA and 4-MIA were observed using reverse transcription PCR amplification. RNA was extracted from 3T3L1 cells using Trizol (AB) solution, and then cDNA was prepared using Quantitect Reverse transcriptase kit (Qiagen), which was then incorporated into dsDNA with oligos synthesized to amplify individual genes as templates. Using the Quantitect SYBR Green PCR kit (AB science) using the characteristics of entering SYBR Green, the amplification process of cDNA was confirmed in real time in 7500-Real-time PCR system (Applied Biosystems, Foster city, CA, USA). The oligos used at this time were found to form a single amplicon of 150-200 bp during PCR amplification, and their nucleotide sequences were as follows.

β3-AR (NM_013462)

forward 5′-AGGCAACCTGCTGGTAATCA-3 ′ (SEQ ID NO 1)

reverse 5′-TCCACAGTTCGCAACCAGTT-3 ′ (SEQ ID NO: 2)

HSL (NM_010719)

forward 5′-TTCGAGGGTGATGAAGGACT-3 ′ (SEQ ID NO: 3)

reverse 5′-ACTCTGGGTCTATGGCGAAT-3 ′ (SEQ ID NO: 4)

adipose TG lipase (ATGL) (NM_025802)

forward 5′-ACCAACACCAGCATCCAGTT-3 ′ (SEQ ID NO: 5)

reverse 5′-TTTGCACATCTCTCGGAGGA-3 ′ (SEQ ID NO: 6)

adiponectin (U49915)

forward 5′-CATGCCGAAGATGACGTTAC-3 ′ (SEQ ID NO 7)

reverse 5′-CGATACACATAAGCGGCTTC-3 ′ (SEQ ID NO: 8)

GAPDH (NM_001001303)

forward 5′-ACATCATCCCTGCATCCACT-3 ′ (SEQ ID NO: 9)

reverse 5′-AGATCCACGACGGACACATT-3 ′ (SEQ ID NO: 10)

The expression level of the gene is expressed as the number of PCR cycles at the point where the cDNA is amplified and the fluorescence reaches the saturation state.

The results are shown in Figures 3-6. 3 to 6 are graphs showing the results of RT-PCR confirming the effect of 1-MIA or 4-MIA on β3-AR, HSL, ATGL, AdipoQ gene expression. Relative fold in the graph is the ratio of the relative expression amount to the expression level of GAPDH. In the figure, # is P <0.05 compared to all cells, ## is P <0.005 compared to all cells, * is P <0.05 compared to adipocytes, ** is P <0.005 compared to adipocytes. As can be seen in Figures 3 to 6, 1-MIA increases the expression level of β3-AR, HSL, ATGL, AdipoQ concentration-dependently, 4-MIA also increases the expression level of β3-AR, HSL, ATGL in a concentration-dependent manner It can be seen that increase. Therefore, it can be seen that 1-MIA and 4-MIA exhibit anti-obesity effect by exothermic reaction and lipolysis through β3-AR, HSL and ATGL, and 1-MIA also shows anti-obesity effect by AdipoQ action. It can be seen.

2-3. HSL and PKA Phosphorylation Measurement

HSL (Hormone sensitive lipase) is present in specific regions of adipocytes, and is an enzyme responsible for the breakdown of fat. Increased activity is known to lead to a decrease in adipose tissue. It is also known that activation of HSL is caused by phosphorylated Protein kinase A (PKA) (see Biochem. Soc. Trans. 31: 1120-1124, 2003; Biochem. J. 364: 73-79, 2002).

Therefore, in this experiment, we tried to confirm the anti-obesity effect through the activation (phosphorylation) experiment of PKA and HSL. 1-MIA and 4-MIA prepared in 2-1. Were dissolved in DMSO, respectively.

Kinase activation was determined by the kinase phosphorylation level. Western blot experiments were performed using antibodies that specifically detect phosphorylated kinase. 3T3L1 cells (American Type Culture Collection) were treated with 1-MIA and 4-MIA at 12.5 μM and 100 μM concentrations for 2 hours, respectively. Proteins were isolated from the cells as follows, and the expression levels of total HSL, phosphorylated-HSL (p-HSL), total PKA, and phosphorylated-PKA (p-PKA) were measured by Western blotting. Cells were collected first, washed twice with cold PBS, and then treated with lysis buffer (10 mM HEPES, 10 mM KCl, 2 mM MgCl ₂ , 1 mM dithiothreitol, 0.1 mM EDTA, 0.1 mM PMSF). After reacting for 30 minutes to melt well, centrifuged at 12000 g (4 degrees Celsius) for 10 minutes to separate intracellular proteins. Proteins are electrophoresed using 10-12% SDS-PAGE and then transferred to PVDF (polyvinylmembrane). Afterwards, 5% skim milk was used to reduce nonspecific reactions, and antibodies (Santa-Cruz Biotechnology, Santacruz, CA) specific to each antigen were reacted at a ratio of 1: 500 to 1: 1000 as the primary antibody. After the treatment, HRP-attached secondary antibodies (anti-rabbit or anti-goat IgG, Santa-Cruz Biotechnology) were treated. Subsequently, the membrane was reacted with an Immobilon Western HRP detection reagent (Millipore, Billerica, MA, USA) and analyzed with a LAS-3000 imaging system (FujiPhotoFilm, Tokyo, Japan). Cells not treated with 1-MIA and 4-MIA were used as controls. The results are shown in Fig. Figure 7 shows Western blot results for pHSL, HSL, pPKA, PKA. As shown in the figure, the treatment of 1-MIA and 4-MIA increased the phosphorylation of HSL, and it can be seen that the phosphorylation of PKA causing the activation of HSL was also increased. Therefore, it can be seen that 1-MIA and 4-MIA can exhibit anti-obesity effect by activating PKA to activate HSL, an enzyme responsible for lipolysis.

Experimental Example 1 Toxicity Test

1-MIA and 4-MIA prepared in the same manner as in 1-1. Were treated to 3T3-L1 cells with varying concentrations, and 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide Cytotoxicity was measured by (MTT) assay (as described in Han et al., J. Agric. Food Cejm 56: 92-98, 2008).

As a result, neither 1-MIA nor 4-MIA showed toxicity at 100 μM. Therefore, it was confirmed that 1-MIA and 4-MIA are safe substances that do not exhibit toxicity to a concentration of 100 μM.

Preparation Example 1 Preparation of Pharmaceutical Composition

1-MIA 20 mg, corn starch 100 mg, lactose 100 mg, magnesium stearate 2 mg prepared in the same manner as in Example 1-1 was filled into gelatin capsules to prepare a capsule.

Preparation Example 2 Preparation of Food Composition

To 1-MIA (0.4 wt%), liquid fructose (0.5 wt%), oligosaccharide (2 wt%), sugar (2 wt%), and salt (0.5 wt%) prepared in the same manner as in Example 1-1. After adding water to adjust the remaining amount, homogeneously blended and sterilized to prepare a health drink.

Preparation Example 3 Preparation of Pharmaceutical Composition

A capsule was prepared by filling 20 mg of 4-MIA, 100 mg of corn starch, 100 mg of lactose, and 2 mg of magnesium stearate in a gelatin capsule prepared in the same manner as in Example 1-1.

Preparation Example 4 Preparation of Food Composition

To 4-MIA (0.4 wt.%), Liquid fructose (0.5 wt.%), Oligosaccharide (2 wt.%), Sugar (2 wt.%), And salt (0.5 wt.%) Prepared in the same manner as in Example 1-1. After adding water to adjust the remaining amount, homogeneously blended and sterilized to prepare a health drink.

<110> KANGHWA COUNTY <120> Composition for anti-obesity <130> P10-096-GHG <160> 10 <170> Kopatentin 1.71 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> sense oligo sequence <400> 1 aggcaacctg ctggtaatca 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> antisense oligo sequence <400> 2 tccacagttc gcaaccagtt 20 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 3 ttcgagggtg atgaaggact 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 4 actctgggtc tatggcgaat 20 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 5 accaacacca gcatccagtt 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 6 tttgcacatc tctcggagga 20 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 7 catgccgaag atgacgttac 20 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 8 cgatacacat aagcggcttc 20 <210> 9 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> forward primer <400> 9 acatcatccc tgcatccact 20 <210> 10 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> reverse primer <400> 10 agatccacga cggacacatt 20

Claims (6)

An anti-obesity composition containing at least one selected from 1-methoxyindole-3-acetonitrile or 4-methoxyindole-3-acetonitrile as an active ingredient. The composition of claim 1, wherein the active ingredient is separated from turnips. The composition of claim 2, wherein the turnip is a fortified turnip. The method of claim 2, wherein the 1-methoxyindole-3-acetonitrile is extracted with turnip with methanol; Partitioning and extracting the methanol extract into ethyl acetate / H ₂ O, and then partitioning and extracting the H ₂ O layer into n-BuOH / H ₂ O; Concentrating the partitioned extracted layer under reduced pressure to obtain an ethyl acetate fraction, n-butanol fraction and water fraction; Silica gel column chromatography on the ethyl acetate fraction (n-hexane-ethyl acetate = 10: 1 → 5: 1 → 3: 1 → 1: 1 → CHCl ₃ -methanol = 10: 1 → 7: 1 → 5: 1 → 3: 1 → 2: 1 → 1: 1) to obtain 18 fractions (BE1 ~ BE18); And the seventh fraction (BE7 fraction) of the 18 fractions is separated from the turnip by a separation method comprising the step of purifying and separating with octadecyl silica gel. The method of claim 2, wherein the 4-methoxyindole-3-acetonitrile is extracted with turnip with methanol; Partitioning and extracting the methanol extract into ethyl acetate / H ₂ O, and then partitioning and extracting the H ₂ O layer into n-BuOH / H ₂ O; Concentrating the partitioned extracted layer under reduced pressure to obtain an ethyl acetate fraction, n-butanol fraction and water fraction; Silica gel column chromatography on the ethyl acetate fraction (n-hexane-ethyl acetate = 10: 1 → 5: 1 → 3: 1 → 1: 1 → CHCl ₃ -methanol = 10: 1 → 7: 1 → 5: 1 → 3: 1 → 2: 1 → 1: 1) to obtain 18 fractions (BE1 ~ BE18); And the ninth fraction (BE9 fraction) of the 18 fractions is separated from the turnip by a separation method comprising the step of purifying and separating with octadecyl silica gel. The composition according to any one of claims 1 to 5, which is a food composition or a pharmaceutical composition.

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