KR20110123002A - Activator for ampk - Google Patents

Abstract

PURPOSE: An AMPK activator containing cinnamic aldehyde is provided to ensure excellent AMPKI activation and to prevent or treat obesity, type 2 diabetes, metabolic syndrome, Alzheimer's disease, and cancer. CONSTITUTION: An AMPK(5' adenosine monophosphate - activator protein kinase) activator contains 1-10 wt% of cinnamic aldehyde as an active ingredient. The AMPK activator is used for preventing or treating obesity, type 2 diabetes, metabolic syndrome, Alzheimer's disease, or cancer. The AMPK activator is a pharmaceutical or food composition. The food composition is manufactured in the form of tablets, granules, powder, and drink by drying the composition with a carrier and capsulating.

Description

AMPM Activator {Activator for AMPK}

The present invention relates to AMPK activators, and more particularly, to AMPK activators effective for the treatment or prevention of obesity, type 2 diabetes, metabolic syndrome, Alzheimer's dementia and cancer.

AMPK activators are substances that increase the activity of AMPK (5'AMP-activated protein kinase; 5 'adenosine monophosphate-activated protein kinase), and AMPK is an enzyme that plays an important role in cellular energy homeostasis. By activating AMPK, it is known that it can have a prophylactic or therapeutic effect such as type 2 diabetes, metabolic syndrome, obesity, cancer, and degenerative disease (dementia), and research on AMPK activators continues. {AMPK and the biochemistry of exercise: Implications for human health and disease. Biochem J 418 (2), 261-275 (2009), Mechanisms linking obesity, chronic kidney disease, and fatty liver disease: the roles of fetuin-A, adiponectin, and AMPK. J Am Soc Nephrol 2010 Mar .; (21) 3: 406-12}.

On the other hand, AMPK activation effect is not known at all with respect to cinnamic aldehyde.

The technical problem to be solved by the present invention is to provide an AMPK activator.

The present invention provides an AMPK (5 'adenosine monophosphate-activated protein kinase; 5' adenosine monophosphate-activated protein kinase) activator containing cinnamic aldehyde as an active ingredient.

The cinnamic aldehyde may be purchased or extracted from the cage.

The cinnamic aldehyde may be included in an amount of 1 to 10% by weight based on the total weight of the activator, there is a fear that the effect is not sufficient at less than 1% by weight, there is a possibility that the effect is no longer greater than 10% by weight.

The AMPK activator is not limited thereto, but may be preferably for treatment or prevention of at least one selected from obesity, type 2 diabetes mellitus, metabolic syndrome, Alzheimer's dementia and cancer.

The AMPK activator may be a pharmaceutical composition or a food 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.

The cinnamic aldehyde exhibits an AMPK activating action.

Therefore, the present invention provides the use of AMPK activation of cinnamic aldehyde.

Also provided are uses for the preparation of AMPK activators of cinnamic aldehydes.

The present invention also provides a method of AMPK activation comprising administering to a mammal (including human) in need of a pharmaceutically effective amount of cinnamic aldehyde.

Unless otherwise stated, the content of the AMPK activator of the present invention is equally applicable to the AMPK activating use, the AMPK activator preparation, and the AMPK activating method of the present invention.

The cinnamic aldehyde and the AMPK activator including the same can be administered orally or parenterally to mammals including humans, and can be administered by formulating an active ingredient 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, gelatin, and the like. It is prepared by adding. 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 cinnamic aldehyde or AMPK activator comprising the same may be administered by subcutaneous injection, intravenous injection or intramuscular injection during parenteral administration.

In the AMPK activator, uses and methods of the present invention, it can be used at a dose of 10-50 mg / kg per adult on the basis of cinnamic aldehyde.

The cinnamic aldehyde can be formulated by adding a pharmaceutically or food-acceptable carrier, excipient or diluent, etc. For formulation, see Remington's Pharmaceutical Science (Recent Edition), Mack Publishing Company, Easton PA et al. Reference may be made.

The AMPK activator of the present invention is excellent in AMPK activating effect, and thus is effective in the prevention or treatment of obesity, type 2 diabetes, metabolic syndrome, Alzheimer's dementia, cancer and the like.

1 is a result of measuring the expression of AMPK protein in adipose tissue after administration of cinnamic aldehyde to the animal by Western blot method.

Example 1 Confirmation of AMPK Activation Effect of Cinnamic Aldehyde

1-1. Preparation of cinnamic aldehyde

Cinnamic aldehyde was purchased from Sigma-Aldrich (St Louis, MO, USA). The cinnamaldehyde was used by dissolving in 0.01% dimethyl sulfoxide (DMSO) in the case of cell experiments, and dissolved in 0.5% sodium carboxymethyl cellulose in the case of animal experiments.

1-2. Preparation and Processing of Cells

3T3-L1 adipocytes were purchased from the American Type Culture Collection (ATCC). Cell culture was performed using Dulbecco's modified Eagle's medium (DMEM) medium (Gibco) containing 10% FBS (fetal bovine serum), penicillin (100 U / ml), streptomycin (100 U / ml), and 37 degrees Celsius in a CO ₂ incubator. Incubated at 5% CO ₂ . 3T3-L1 preadipocytes were treated with differentiation-inducing media (5% fetal bovine serum, 0.5 mM isobutylmethylxanthine, 1 mM dexamethasone and DMEM with 10 μg / mL insulin) and cinnamic aldehyde (20 μM, 40 μM) After exchange and incubation for a total of 4 days, and further exchanged with DMEM containing 10 μg / mL insulin was cultured for 2 more days. Thereafter, two days of incubation with DMEM containing only FBS induced differentiation for a total of 8 days.

1-3. AMPK activation effect confirmed

Using the adipocytes prepared in the above 1-2. Was tested to confirm the AMPK activation effect by Western blot method.

Adipose cells were homogenized with lysis buffer for protein analysis. Protein quantification was measured using a Bio Rad assay reagent (Bio-Rad, USA), and 20 μg of the quantified protein was isolated by 8% SDS-PAGE. Afterwards, the gel was transferred to a membrane (Milipore, Cat.No: IPVH00010), blocked with 5% skim milk for 1 hour at room temperature, and diluted with a 1: 1000 ratio of primary antibody (P-AMPK, Cell Signaling, 2531; AMPK, Cell). Signaling, 2532; P-ACC, Cell Signaling, 3661; ACC, Cell Signaling, 3662) was reacted overnight at 4 ° C. After washing three times with tris-buffered saline tween-20 (TBST), the mixture was reacted with secondary antibody (Santa Cruz, sc-2313) diluted at a ratio of 2000 to 1 hour at room temperature. After washing 3 times with TBST and using the ECL solution (Amersham, Sweden) was developed on the X-ray film.

The results are shown in FIG. 1 is a diagram showing the results of confirming the expression of the activity of AMPK and ACC by Western blot method. 1, DM means differentiation induction medium, CA means cinnamic aldehyde,-means not treated with the corresponding component, + means treated, each number means a concentration (uM). In addition, pAMPK means the amount of phosphorylated AMPK protein, AMPK means the amount of AMPK protein of the base level. Similarly, pACC means the amount of phosphorylated ACC protein, ACC means the amount of ACC protein at base level, and Actin means control protein as loading control.

From the above results, it can be seen that cinnamic aldehyde increases the phosphorylation of AMPK in a concentration-dependent manner. As a result, it can be seen that cinnamaldehyde activates AMPK. In addition, Acetyl-CoA carboxylase (ACC) is one of the enzymes produced when fat metabolism is increased to inhibit further use of ATP by inhibiting key enzymes such as ACC in fatty acid synthesis by activating the AMPK by cinnamicaldehyde. It can be seen that it is effective in the treatment or prevention of obesity, metabolic syndrome, type 2 diabetes, Alzheimer's dementia, cancer and the like.

1-4. Preparation and Processing of Animals

Five-week-old male Imprinting Control Region (ICR) mice (Orient Co., Ltd.) were adapted for one week before being used for the experiment. The dietary methods of overfeeding and energy dense diets were provided and transformed into a diet-induced obese (DIO) model that induced obesity. The experimental group was the control group provided with the normal diet (10% Kcal% fat, Research DIETS), the high-fat diet group fed high-fat diet (45% Kcal% fat, Research DIETS), the high-fat diet and Example 1-1. Example 1-1 (20mg / kg) administered with cinnamic aldehyde (20mg / kg), high fat diet and Example 1-1 (40mg) administered with cinnamic aldehyde (40mg / kg) of Example 1-1. / kg) divided into administration groups. Cinnamic aldehyde of Example 1-1 was administered orally once a day for two weeks, body weight and blood glucose were measured weekly, and blood insulin, triglycerides, total cholesterol, HDL-cholesterol, and plasma nonesterified fatty acid (NEFA) It was measured at the end of the experiment. Cinnamic aldehyde was dissolved in 0.5% CMC (carboxymethylcellulose) when administered.

Body weight was measured with a scale (OHAUS, Adeventurer Pro AVG812), blood glucose was measured with a SMARTLAB (Erba, USA) instrument. Blood insulin, triglyceride (TG), total cholesterol (TC), HDL-cholesterol (HDL-cholesterol), plasma nonesterified fatty acid (NEFA), glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), blood urea Nitrogen) was measured by ELISA reader (Labsystems, Finland) assay by collecting blood from the heart and centrifuging at 4 ℃ serum. In addition, homeostasis model assessment of insulin resistance (HOMA-IR), an insulin resistance index, was calculated. The calculation method is Insulin (μU / mL) x glucose (mM) /22.5.

Statistical analysis of data is performed p <0.05, p <0.01, p < one won are distributed analysis at 0.001 level (one-way analysis of variance) with a mean ± standard deviation calculated using the SPSS 14.0 statistics program and LSD (least Significant difference test) was used to test the significance between the mean of each group.

1-5. Anti-obesity effect confirmed

Weight measurement results for the mice treated in 1-4. Are summarized in the following table.

TABLE 1

As shown in the table, the high-fat diet group can be confirmed that the weight gain is significant compared to the control group induced obesity. When cinnamic aldehyde was administered under the same conditions as the high fat diet group, it can be seen that the weight gain decreases depending on the concentration of cinnamic aldehyde. Therefore, it can be seen that cinnamic aldehyde has an anti-obesity effect.

1-6. Confirm antidiabetic effect

The blood glucose and blood insulin measurement results for the mice treated in 1-4. Are summarized in the following table.

TABLE 2

As shown in the above table, the high-fat diet group showed a higher concentration of blood sugar and insulin than the control group compared to the control group, but it can be seen that the concentration of the cinnamaldehyde administration group is concentration-dependently reduced. In addition, it can be seen that the HOMA-IR value is also reduced compared to the Cinnamic aldehyde-treated group high-fat diet group.

Therefore, it can be seen that cinnamic aldehyde is effective for the treatment or prevention of type 2 diabetes, metabolic syndrome, Alzheimer's dementia, etc. by reducing blood sugar, decreasing blood insulin concentration, improving insulin sensitivity, and the like.

1-7. Blood Indicator Analysis

NEFA, blood glucose and blood insulin measurement results for the mice treated in 1-4. Are summarized in the following table.

[Table 3]

As shown in the above table, the cholesterol related to hyperlipidemia, arteriosclerosis, etc. is increased in the high fat diet group compared to the control group, it can be seen that the tendency of concentration-dependent decrease by the administration of cinnamic aldehyde.

In addition, in the case of triglyceride-associated free fatty acid NEFA in the high fat diet group compared to the control group, it can be seen that the tendency of concentration-dependent decrease by the administration of cinnamic aldehyde.

Therefore, cinnamic aldehydes show the activation of AMPK, a protein that corresponds to the metabolic sensor, and the resulting fatty acid oxidation-promoting blood glucose levels and lipid profile, resulting in associated obesity, type 2 diabetes, metabolic syndrome, Alzheimer's dementia and cancer. It can be seen that it is effective in the treatment or prevention of the back.

Preparation Example 1 Preparation of Pharmaceutical Composition

20 mg of cinnamic aldehyde of Example 1-1, 100 mg of corn starch, 100 mg of lactose, and 2 mg of magnesium stearate were filled into gelatin capsules to prepare a capsule.

Preparation Example 2 Preparation of Food Composition

Cinnamic aldehyde (4 wt%), liquid fructose (0.5 wt%), oligosaccharide (2 wt%), sugar (2 wt%), and salt (0.5 wt%) of Example 1-1 were added to the remaining amount. After adjusting, homogeneously blended and sterilized to prepare a health drink.

Claims (3)

AMPK (5 'adenosine monophosphate-activated protein kinase) activator containing cinnamic aldehyde as an active ingredient. The AMPK activator according to claim 1, which is for treatment or prevention of at least one selected from obesity, type 2 diabetes, metabolic syndrome, Alzheimer's dementia and cancer. The AMPK activator according to any one of claims 1 to 2, which is a pharmaceutical composition or a food composition.

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