KR101877443B1 - Composition for preventing, improving or treating obesity comprising Loganic acid or its derivatives - Google Patents

Abstract

The present invention relates to a composition for preventing, improving or treating obesity, which contains a loganic acid and derivatives thereof or pharmaceutically allowable salts thereof as effective ingredients. The loganic acid in the present invention has excellent efficacy in inhibiting fat cell differentiation, inhibiting fat cell differentiation, and activating marker expression, and has efficacy in effectively inhibiting weight gain in a menopausal obese animal model, fat accumulation of the liver and fat cell size increase of abdominal adipose tissues. Therefore, the loganic acid and the composition containing the same as the effective ingredient can be usefully utilized as a pharmaceutical agent for preventing, improving and treating the obesity.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a composition for preventing, improving or treating obesity, which comprises rosiglucanic acid or a derivative thereof as an active ingredient.

The present invention relates to a composition for preventing, ameliorating or treating obesity, which comprises Loganic acid, a derivative thereof or a pharmaceutically acceptable salt thereof as an active ingredient derived from a gangrene extract.

In 2014, at least one in three adults over the age of 18 has exceeded their standard weight, and one out of every 10 is obese. In addition, abdominal obesity and visceral fat accumulation are the greatest risk factors of adult diseases, and the socioeconomic loss due to obesity is enormous.

As obesity became a global concern, the World Health Organization (WHO) declared the war on obesity on May 22, 2004. At the 57th annual meeting of the World Health Organization, which was closed in Geneva, the ministers of each country unanimously passed the 'global strategy for diet, exercise and health'. WHO says over one billion people worldwide are overweight and at least 300 million are obese. Also, by 2020, deaths from non-communicable diseases are expected to account for 73% of the total and 60% of the costs of treatment. Non-communicable diseases include cardiovascular disease, diabetes and cancer, and are caused mainly by hypertension, cholesterol, eating less of fruits and vegetables, overweight and obesity, lack of exercise and tobacco, WHO said. The World Health Organization (WHO) recommended that governments share the severity of obesity and establish an efficient and integrated agriculture and health policy to educate the public. WHO's World Strategy for Diet, Exercise, and Health recommended a balanced diet, proper exercise, reduced consumption of fat, salt and sugar, and plenty of fruits and vegetables. But there was also controversy among the member countries. The sugar producer, Brazil, agreed with the WHO global strategy, citing the sugar-related provision should not be regarded as a way to distort trade in any way.

PCT International Publication No. WO2015-198346 (published Dec. 30, 2015)

It is an object of the present invention to provide a composition for preventing, improving or treating obesity, which comprises Loganic acid, a derivative thereof or a pharmaceutically acceptable salt thereof as an active ingredient, There is good efficacy in the inhibition of differentiation in pre-adipocyte (3T3-L1), and weight loss and abdominal visceral fat decrease in ovariectomized obesity model. And to provide a composition for the treatment of cancer.

The present invention provides a pharmaceutical composition for preventing or treating obesity, which comprises Loganic acid, a derivative thereof or a pharmaceutically acceptable salt thereof as an active ingredient, and a health food composition for preventing or ameliorating obesity.

In addition, the present invention provides a reagent composition for inhibiting the expression of an adipocyte activation marker gene, which comprises Loganic acid, a derivative thereof, or a pharmaceutically acceptable salt thereof as an active ingredient, in vitro.

The present invention also relates to a method for inhibiting the in vitro expression of an adipocyte activating marker gene, which comprises treating adipocytes with a loganic acid, a derivative thereof or a pharmaceutically acceptable salt thereof in vitro, ≪ / RTI >

The present invention relates to a composition for preventing, improving or treating obesity, which comprises Loganic acid, a derivative thereof or a pharmaceutically acceptable salt thereof as an active ingredient. The rognic acid of the present invention inhibits adipocyte differentiation Adiponectin (Adipoq), peroxisome proliferator-activated receptor gamma (Ppar gamma), lipoprotein lipase (Lipase) and lipoprotein lipase (Lip), perilipin1 (Plin1), fatty acid binding protein 4 (Fabp4), glucose transporter type 4 (Slc2a4), CCAAT / enhancer-binding protein alpha CCAAT / enhancer-binding protein alpha (Cebpa) gene. In addition, the ovariectomized obesity model results in weight loss and reduction of visceral fat mass, ultimately preventing or treating obesity. Therefore, it is expected that the rhodonic acid of the present invention and the composition containing it as an active ingredient can be usefully used as a pharmaceutical preparation for prevention, improvement and treatment of obesity.

Figure 1 shows the chemical structure of Loganic acid.
FIG. 2 shows the results of treatment of 3T3-L1 cells, pre-adipocytes, with a concentration of 2 μg / mL, 10 μg / mL and 50 μg / mL, This is a schematic diagram showing inhibition of differentiation into adipocytes.
FIG. 3 shows cytotoxicity of 3T3-L1 cells, which are pre-adipocytes, after treatment with rogavinic acid at a concentration of 2 μg / mL, 10 μg / mL and 50 μg / mL, It is a schematic diagram showing that the acid is not cytotoxic.
Figure 4 shows that 3T3-L1 cells, a pre-adipocyte, were treated with 2 μg / ml, 10 μg / ml and 50 μg / ml of rogavinic acid, Adiponectin (adiponectin), peroxisome proliferator-activated receptor gamma (Ppar gamma), lipoprotein lipase (Lpl), perilipin 1 (Plin 1), fatty acid binding MRNA expression level of protein 4 (fatty acid binding protein 4 (Fabp4), glucose transporter type 4 (Slc2a4), CCAAT / enhancer-binding protein alpha (Cebpa) Which is a schematic diagram showing the effect of suppressing the expression of transcription factors involved in the adipocyte formation of roganic acid. Induction: Treatment with adipocyte differentiation inducer (IBMX, Dexa, Insulin: MDI), * p <0.05 vs. Induction only.
FIG. 5 shows the body weight (FIG. 5a) of the obese mouse model (ovariectomized mouse) after 12 weeks of administration of rognacic acid at 2, 10 and 50 mg / kg / day, (Fig. 5c), and H & E staining showed that it was significantly effective in inhibiting lipid accumulation in liver liver and increase of fat cell size in abdominal adipose tissue. Sham: normal group (laparotomy, suture); OVX: ovariectomized group (open, ovariectomy, suture); E2: estradiol treated group (positive control group); LA: Loganic acid treated group (2, 10, 50 mg / kg / day); * p < OVX.

Thus, the present inventors confirmed the in vivo efficacy of rogannic acid, a single compound effective for weight gain and improvement of fat accumulation and fat cell size increase in liver and abdominal adipose tissue, and completed the present invention.

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

Specifically, the rosigonic acid may be derived from a safflower extract, but is not limited thereto.

In particular, the pharmaceutical composition may inhibit adipocyte differentiation and may be adiponectin (Adipoq), peroxisome proliferator-activated receptor gamma (Ppar gamma), lipoprotein lipase Lip), perilipin1 (Plin1), fatty acid binding protein 4 (Fabp4), glucose transporter type 4 (Slc2a4) and CCAAT / enhancer-binding protein alpha / enhancer-binding protein alpha (Cebpa), but is not limited thereto.

The loganic acid of the present invention is represented by the formula (1), and the chemical formula thereof is C ₁₆ H ₂₄ O ₁₀ .

&Lt; Formula 1 >

In the present invention, the pharmaceutically acceptable salts include salts with organic acids selected from the group consisting of oxalic acid, maleic acid, fumaric acid, malic acid, tartaric acid, citric acid and benzoic acid, or inorganic acids selected from the group consisting of hydrochloric acid, sulfuric acid, phosphoric acid and hydrobromic acid May be a salt.

When the composition of the present invention is a pharmaceutical composition, the pharmaceutical composition may be formulated into a cream, a gel, a patch, a spray, an ointment, a warning agent, a lotion, a liniment, a pasta agent and a cataplasma agent. Meanwhile, the pharmaceutical composition may contain a pharmaceutically acceptable carrier in addition to the above-mentioned Loganic acid. Such pharmaceutically acceptable carriers are those conventionally used in pharmaceutical preparations, and include lactose, dextrose , Sucrose, mannitol, starch, acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, Propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like, but are not limited thereto. In addition, the pharmaceutical composition may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. as an additive.

The pharmaceutical composition may be administered according to the severity of obesity. Usually, local administration is preferred. The dosage of the active ingredient in the pharmaceutical composition may vary depending on the route of administration, the severity of the disease, the age, sex, and weight of the patient, and may be administered once to several times per day.

The present invention also provides a health food composition for preventing or ameliorating obesity comprising Loganic acid, a derivative thereof or a pharmaceutically acceptable salt thereof as an active ingredient.

The health food composition may be provided in the form of powder, granule, tablet, capsule, syrup, beverage or ring, and the health food composition may contain food or food additives other than Loganic acid according to the present invention, And can be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to its use purpose, for example, prevention, health or therapeutic treatment.

The effective dose of Loganic acid contained in the above health food composition can be used in accordance with the effective dose of the above pharmaceutical composition but can be used for health and hygiene purposes or for long- May be less than the above range, and since the active ingredient has no problem in terms of safety, it can be used in an amount exceeding the above range.

There is no particular limitation on the type of the health food, and examples thereof include meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, Drinks, alcoholic beverages and vitamin complexes.

The present invention also relates to a pharmaceutical composition comprising Loganic acid, a derivative thereof or a pharmaceutically acceptable salt thereof as an active ingredient and further comprising at least one of adiponectin (Adipoq), peroxisome proliferator-activated receptor gamma (Ppar?), lipoprotein lipase (Lpl), perilipin1 (Plin1), fatty acid binding protein 4 (Fabp4), glucose transporter type 4 The present invention provides a reagent composition for inhibiting the expression of at least one adipocyte activation marker gene selected from the group consisting of S1a1, S1c2a4, and CCAAT / enhancer-binding protein alpha (Cebpa) .

The present invention also relates to a pharmaceutical composition comprising adiponectin (Adipoq), peroxyacetic acid (Adipoq), and the like, which comprises the step of treating adipocytes with a loganic acid, a derivative thereof or a pharmaceutically acceptable salt thereof, (PPLγ), lipoprotein lipase (Lpl), perilipin 1 (Plin 1), fatty acid binding protein 4 (Fabp 4), and lipoprotein lipase In vitro expression of one or more adipocyte activation marker genes selected from the group consisting of glucose transporter type 4 (Slc2a4) and CCAAT / enhancer-binding protein alpha (Cebpa) Lt; / RTI &gt;

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are intended to illustrate the contents of the present invention, but the scope of the present invention is not limited to the following examples. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

< Example  1> Of Loganic acid  Chemical structure

The single test compound used in the animal experiments in the present invention is loganic acid. Roganic acid has a chemical structure of C ₁₆ H ₂₄ O ₁₀ and is a naturally occurring single compound that is abundant in Gentiana scabra (Fig. 1).

< Example  2> Rogainic acid  Inhibition of adipocyte differentiation

The efficacy of rogannic acid for the differentiation of adipocytes was assessed using 3T3-L1 cells. 3T3-L1 cells, a pre-adipocyte, are committed stem cells that differentiate into adipocytes when treated with differentiation-inducing agents. After treatment with 3 different compounds [MDI: 3-Isobutyl-1-methylxanthine (IBMX) 0.5 mM / mL, Dexamethasone (Dexa) 1 μM / mL, Insulin 10 μg / mL] The effects of loganic acid on proliferation and differentiation were examined.

3T3-L1 cells were treated with adipocyte inducer (MDI) and treated at the same time with rogaconic acid, and the degree of differentiation was analyzed after 8 days of cell culture. The rosiglassic acid was dissolved in an aqueous 1% dimethylsulfoxide (DMSO) solution to a concentration of 2 μg / mL (LA2), 10 μg / mL (LA10) and 50 μg / mL (LA50) Cells were cultured in the same volume of 1% DMSO solution in the absence of saccharinic acid treatment. The degree of adipocyte differentiation activity was measured by Oil Red O staining. Specifically, cells are washed with physiological saline and fixed with 4% paraformaldehyde to the bottom of the cell culture plate well. Fixed cells are stained with Oil Red O reagent for 2 hours and then confirmed by microscopy (Fig. 2).

Cytotoxicity of rogannic acid was assessed by cell proliferation analysis using the EZ-Cytox Enhanced Cell Viability Assay Kit product using a water-soluble tetrazolium salt. Specifically, 3T3-L1 cells (3 × 10 ³ cells / well) as lipid precursor cells were cultured in a medium supplemented with 10% fetal bovine serum, 1 mM sodium pyruvate, 100 unit / L penicillin and 100 mg / L streptomycin And cultured at 37 캜 in which 5% CO ₂ was supplied. The adipocyte differentiation inducer (MDI) and rognacic acid were co-treated with 3 concentrations (2 μg / mL, 10 μg / mL, 50 μg / mL) and cultured for 8 days. Tetrazolium salt (Tetrazolium salt) was cultured in each cultured cell and sample, incubated at 37 ° C for 2 hours, and absorbance was measured at 450 nm. Compared with the control group treated with physiological saline, in all the experimental groups treated with rogavinic acid at a concentration of 2 μg / mL (LA2), 10 μg / mL (LA10) and 50 μg / mL (LA50) L1 cell proliferation (Fig. 3).

Next, the adipocyte differentiation inducer (MDI) and rosiglucan were co-treated with 3 concentrations (2 μg / mL, 10 μg / mL, 50 μg / mL) and cultured for 8 days. The control (Control) did not treat the differentiation inducer. Adiponectin (Adipoq), peroxisome proliferator-activated receptor gamma (Ppar gamma), lipoprotein lipase (Lpl), perilipin 1 , Plin1, fatty acid binding protein 4 (Fabp4), glucose transporter type 4 (Slc2a4), CCAAT / enhancer-binding protein alpha (Cebpa) ) MRNA expression level of the gene was analyzed. In the cells not treated with rosiglucan, the expression level of adipocyte differentiation and adipocyte activation marker gene was markedly increased by treatment with adipogenic differentiation inducer, but the expression level of these markers was significantly increased in rognacic acid treated cells (Fig. 4). Therefore, rogannic acid seems to be effective in inhibiting obesity by inhibiting the expression of genes involved in adipocyte differentiation and adipocyte activation.

< Example  3> Inhibitory effect of roganic acid on weight gain, inhibition of hepatic fat accumulation, and inhibition of abdominal adipocyte growth in mouse model of obesity

In general, when estrogen is reduced in female menopause, new estrogen is produced from adipocytes to offset the estrogen deficiency. In this process, more fat cells are produced, which can cause abdominal obesity. That is, fat accumulation in the liver and fat cell size increase in the abdominal adipose tissue are caused by the increase of the insulin and leptin, which are hormone proteins related to the lipid metabolism.

A 10-week-old ovariectomized-mouse (OVX mouse) was used as an animal model for menopausal obesity in order to evaluate the in vivo efficacy of rogainic acid (ovariectomized-mouse, OVX mouse) ). OVX mouse group (negative control group) administered only with physiological saline to the ovariectomized mice, estradiol (estradiol), which is an obesity-improving compound, in the ovariectomized mice E2) was dissolved in sesame oil and injected intramuscularly at a dose of 36 μg / kg / day. E2 A mouse group (positive control) was used as a control. In the experimental group, oral doses of rogavinic acid were given at doses of 2 mg / kg / day (LA2), 10 mg / kg / day (LA10) and 50 mg / kg / day (LA50). 10-week-old sham-operated and ovariectomized ddY female mice were purchased from the Central Laboratory Animals and transferred to a clean animal breeding area after a 1-week purification period in the laboratory animal quarantine room. The weight of each mouse was measured and group separation was performed so that there was no statistically significant weight difference between the experimental groups. The rosiglass used for the experiment was dissolved in 1% DMSO solution to make the test solution. To carry out the laboratory animal center, it was commissioned by Soya Green Tech Co., Ltd., which is a radiation irradiation company, and sterilized by gamma irradiation.

After 12 weeks of administration of rogainic acid in a male model of obesity, obesity and abdominal adipose tissue were removed and histological staining was performed. Compared with normal ovariectomized Shame mice, ovariectomized OVX mice showed a significant increase in body weight and fat accumulation in the liver (Fig. 5a, Fig. 5b), and the fat cells of the abdominal fat tissue were significantly (Fig. 5C). However, in the roganic acid treated group, the fat accumulation in the body and liver and the fat cell size increase in the abdominal adipose tissue were significantly suppressed than the OVX obese model. In addition, weight increase, liver fat accumulation and fat cell size in the estradiol administration-positive control group were markedly decreased as compared with OVX mice (FIGS. 5A, 5B and 5C).

Claims (8)

delete delete delete delete delete delete The present invention relates to a pharmaceutical composition containing Loganic acid or a pharmaceutically acceptable salt thereof as an active ingredient and is useful as an adiponectin (Adipoq), a peroxisome proliferator-activated receptor gamma (Ppar gamma) Lipoprotein lipase (Lpl), perilipin1 (Plin1), fatty acid binding protein 4 (Fabp4), glucose transporter type 4 (Slc2a4) and CCAAT / enhancer- Wherein the reagent composition inhibits expression of at least one adipocyte activation marker gene selected from the group consisting of CCAAT / enhancer-binding protein alpha (Cebpa). (Adiponectin), peroxisome (Adipoq), and peroxisome proliferator-activated receptor gamma, including the step of treating adipocytes with loganic acid or a pharmaceutically acceptable salt thereof in vitro. lipoprotein lipase (Lpl), perilipin 1 (Plin 1), fatty acid binding protein 4 (Fabp 4), glucose transporter 4 (glucose transporter 4) type 4; Slc2a4) and the CCAAT / enhancer-binding protein alpha (Cebpa).

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