KR101598380B1 - Composition for prevention or treatment of diabetes or diabetes complication - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the prevention or treatment of diabetes or diabetic complications and to a health functional food for improvement, and more particularly to a pharmaceutical composition for prevention or treatment of diabetes or diabetic complications having excellent antidiabetic effect including N-phenylbenzamide derivatives or salts thereof And to a health functional food.

Description

Technical Field [0001] The present invention relates to a composition for preventing or treating diabetes or diabetic complications,

The present invention relates to a composition for preventing or treating diabetes or diabetic complications.

Diabetes mellitus is a type of metabolic disorder caused by lack of insulin secreted by the pancreas or by its inability to function, which causes the concentration of glucose present in the blood to increase, which exceeds the resorptive capacity of the tubule, .

As the diabetes continues to be chronic, it causes not only the metabolism of carbohydrates but also the metabolism of lipids and proteins, which causes complications such as retinas, kidneys, nerves and cardiovascular diseases.

In modern times, dietary changes and lack of exercise have caused a great change in the energy metabolism of the human body. Diabetes globally is one of the major causes of adult death. According to a report by the World Health Organization (WHO), there are about 1.9 billion diabetics worldwide in 2009, and by 2030 it is expected to increase to about 370 million.

AMP-activated protein kinase (AMPK), which is activated by external stimuli such as exercise, blocks the pathway of fatty acid synthesis that consumes energy and produces energy Energizes the pathway of blood glucose absorption. Akt, which regulates insulin sensitivity, enhances cell survival and promotes the intracellular delivery of glucose. Acetyl CoA carboxylase (ACC) is a protein that produces energy, Surplus glucose is involved in the conversion to fat. In addition, PPAR-gamma (Peroxisome proliferator-activated receptor-gamma.) Is a receptor that increases the expression of genes involved in glucose metabolism in the nucleus. Activation of PPAR-gamma causes hypoglycemia.

On the other hand, Korean Patent Publication No. 2005-0120773 discloses a drug for treating diabetes comprising a sulfonylurea compound as a drug for treating diabetes. Sulfonylurea drugs promote the secretion of insulin made in pancreatic β cells. However, the sulfonylurea system has a problem that side effects such as hypoglycemia, digestive disorders, tachycardia, headache, and hepatotoxicity can be induced.

United States Patent No. 8,247,407, United States Patent No. 6,291,495 and Korean Patent No. 867,485 disclose an acetylenic compound. Biguanide (metformin) drugs regulate blood sugar by inhibiting sugar production in the liver. Since the bguyuanide system does not cause hypoglycemia unlike the sulfonylurea system, it is used alone in patients with mild diabetes or when the sulfonylurea system fails to be prescribed, it is prescribed in combination with the sulfonylurea system. However, rarely, lactic acidosis can occur, and anorexia, nausea, abdominal discomfort, and diarrhea can occur.

European Patent Publication No. 2007-0708206 and International Patent Publication No. 2013-115740 disclose drugs for the treatment of diabetes comprising meglitinide derivatives. The meglitinide drug is a non-sulfonylurea system and binds to the sulfone urea receptor of pancreatic ß cells to induce insulin secretion, which is quicker and less effective in reducing blood glucose levels than sulfonylurea. However, there is a problem that slight hypoglycemia can be induced.

In addition, glucagon-like peptides (GLP-1 analog) have been reported to have adverse effects such as hypoglycemia, gastrointestinal disorders, headache, abdominal pain and the like. The α-glucosidase inhibitor has been shown to exert a reduced digestive function and amylin analog Hypoglycemia, and nausea have been reported.

There is no blood glucose control agent that can be safely used with excellent efficacy in comparison with the seriousness of diabetes. It is urgent to develop a therapeutic agent having an excellent antidiabetic effect while being safely ingested in the long term.

It is an object of the present invention to provide a pharmaceutical composition and a health functional food for preventing or treating diabetes mellitus or diabetic complication with excellent antidiabetic effect.

The present invention provides a pharmaceutical composition for preventing or treating diabetes mellitus or diabetic complications comprising the compound of the formula (1) or a salt thereof.

[Chemical Formula 1]

Wherein the substituents R ₁ , R ₂ and R ₃ are independently selected from an alkoxy group having 1 to 3 carbon atoms.

Preferably, the compound of formula (1) may be a compound of formula (2).

(2)

(Wherein the substituents R ₄ , R ₅ and R ₆ are independently selected from an alkoxy group having 1 to 3 carbon atoms)

More preferably, the compound of formula (1) may be a compound of formula (3).

 (3)

In order to accomplish still another object of the present invention, the present invention provides a health functional food for preventing or ameliorating diabetes or diabetic complications comprising a compound of the following general formula (1) or a salt thereof.

[Chemical Formula 1]

Wherein the substituents R ₁ , R ₂ and R ₃ are independently selected from an alkoxy group having 1 to 3 carbon atoms.

Preferably, the compound of formula (1) may be a compound of formula (2).

(2)

(Wherein the substituents R ₄ , R ₅ and R ₆ are independently selected from an alkoxy group having 1 to 3 carbon atoms)

More preferably, the compound of formula (1) may be a compound of formula (3).

 (3)

The present invention relates to a pharmaceutical composition capable of promoting the transcription activity of PPAR-gamma which is a target molecule of diabetes prevention and exhibiting an excellent glucose absorption effect in muscle cells to prevent or treat diabetic or diabetic complications and to prevent or ameliorate diabetic or diabetic complications It can be used as a health functional food.

1 is a graph showing the effect of 2,5-dimethoxy-N- (4-methoxyphenyl) benzamide on PPAR-gamma transcription activity according to an embodiment of the present invention.
2 is a graph showing the effect of 2,5-dimethoxy-N- (4-methoxyphenyl) benzamide on glucose uptake of C2C12 cells according to an embodiment of the present invention.
3 is a graph showing the effect of 2,5-dimethoxy-N- (4-methoxyphenyl) benzamide on expression of p-ACC, p-AMPK and p-Akt in C2C12 cells according to an embodiment of the present invention Fig.
4 is a graph showing the effect of 2,5-dimethoxy-N- (4-methoxyphenyl) benzamide on the amount of p-AMPK expression in C2C12 cells according to an embodiment of the present invention.
5 is a graph showing the effect of 2,5-dimethoxy-N- (4-methoxyphenyl) benzamide on the amount of p-ACC expression in C2C12 cells according to an embodiment of the present invention.

Hereinafter, the present invention will be described in detail. However, it should be understood that the present invention may be embodied in many other specific forms without departing from the spirit or essential characteristics thereof.

In one embodiment, the invention relates to a pharmaceutical composition for the prophylaxis or treatment of diabetes or diabetic complications comprising a compound of formula (I) or a salt thereof.

[Chemical Formula 1]

Wherein the substituents R ₁ , R ₂ and R ₃ are independently selected from an alkoxy group having 1 to 3 carbon atoms and may be the same or different from each other. Specifically, the alkoxy having 1 to 3 carbon atoms means any one of methoxy, ethoxy, n-propoxy and i-propoxy, and R ₁ , R ₂ and R ₃ are preferably methoxy.

Preferably, formula (1) may be the following formula (2).

(2)

Wherein the substituents R ₄ , R ₅ and R ₆ are independently selected from an alkoxy group having 1 to 3 carbon atoms and may be the same or different from each other. Specifically, the alkoxy having 1 to 3 carbon atoms means any one of methoxy, ethoxy, n-propoxy and i-propoxy.

More preferably, formula (1) may be the following formula (3).

(3)

The pharmaceutical composition according to the present invention may be formulated into oral compositions such as powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, external preparations, suppository syringes, pre-filled syringes ) Solution form, or a lyophilized form, but the present invention is not limited thereto.

In the case of formulation, it may be prepared using diluents or excipients such as fillers, extenders, binders, humectants, disintegrants, surfactants and the like which are usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one or more excipients such as starch, calcium carbonate, sucrose, lactose, gelatin, . In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used.

Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups, and the like. Various excipients such as wetting agents, sweetening agents, fragrances, preservatives and the like are included in addition to water and liquid paraffin which are conventionally used simple diluents . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, suppositories, and the like.

The preferred dosage of the pharmaceutical composition of the present invention can be appropriately selected depending on the condition and body weight of the patient, the degree of obesity, the type of disease, the drug form, the administration route and the period. It is preferable for the composition of the present invention that the active ingredient is administered at 0.2 to 200 mg / kg per day for optimal efficacy. The administration may be carried out once a day or divided into several doses, but is not limited thereto.

The pharmaceutical composition of the present invention can be used for prevention or treatment of diabetes. In addition, the pharmaceutical composition of the present invention can be used for preventing or treating complications related to diabetes or diabetes.

The type of the diabetic complication is not particularly limited and it is classified into a group consisting of wound, stroke, myocardial infarction, skin infection, skin aging, neuropathy, lens opacity, cataract, retinopathy, nephropathy, cardiomyopathy, platelet aggregation, endothelial dysfunction, , And preferably one or more selected from the group consisting of wounds, skin aging, and skin infections.

In another embodiment, the invention relates to a health functional food for the prevention or amelioration of diabetes or diabetic complications comprising a compound of the formula (I) or a salt thereof.

[Chemical Formula 1]

Wherein the substituents R ₁ , R ₂ and R ₃ are located on the benzene ring and are independently selected from alkoxy groups having 1 to 3 carbon atoms and may be the same or different from each other. Specifically, the alkoxy having 1 to 3 carbon atoms means any one of methoxy, ethoxy, n-propoxy and i-propoxy, and is preferably methoxy.

Preferably, the compound of formula (1) is a health functional food for prevention or amelioration of the following formula (2).

(2)

Wherein the substituents R ₄ , R ₅ and R _6, which are located on the benzene ring and are independently selected from alkoxy groups having 1 to 3 carbon atoms, may be the same or different from each other. Specifically, the alkoxy having 1 to 3 carbon atoms means any one of methoxy, ethoxy, n-propoxy and i-propoxy.

More preferably, formula (1) may be the following formula (3).

 (3)

The health functional food of the present invention can be manufactured in various forms such as, but not limited to, various drinks, gum, tea, confectionery, vitamin complex, health supplement and the like.

The preferred intake amount of the health functional food of the present invention varies depending on the condition and body weight of the recipient, the degree of obesity, the type of food, the period of consumption, and can be appropriately selected. It is preferable for the composition of the present invention that the active ingredient is ingested 0.2 to 200 mg / kg per day for optimal effect.

The compounds of the formulas (1) to (3) according to the present invention can be prepared by a conventional chemical synthesis method, and there is no particular limitation on the synthesis method.

The compounds of the formulas (1) to (3) according to the present invention can be used in the form of a salt, and the kind of the salt is not particularly limited. Specifically, acid addition salts formed with free acids such as methanesulfonate, ethanesulfonate, fumarate, succinate, hydrochloride, citrate, maleate, tartrate and hydrobromide, formed with hydrochloric acid, phosphoric acid and sulfuric acid Salts may also be used and also salts such as sodium, alkaline earth metal salts, calcium, magnesium, triethylamine, morpholine, N-methylpiperidine, N-ethylpiperidine, procaine, dibenzylamine, Dibenzylethylamine, tris- (2-hydroxyethyl) amine, N-methyl d-glucamine or lysine. A preferred pharmaceutically acceptable salt is a sodium salt.

It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the present invention. Such variations and modifications are intended to be within the scope of the appended claims.

Manufacturing example  - 2,5- Dimethoxy -N- (4- Methoxyphenyl ) Benzamide

According to one embodiment of the present invention, 2,5-dimethoxy-N- (4-methoxyphenyl) benzamide is synthesized according to Biochemical and Biophysical Research Communications (2006) 349, 39-49, Synthesis, discovery and mechanism of 2, 6-dimethoxy-N- (4-methoxyphenyl) benzamide as potent depigmenting agent in the skin (Choi SY, Hwang JS, Kim S, Kim SY.).

The C2C12 muscle cells used in the examples were purchased from American Type Culture Collection (US), the Phospho-AMPK Thr172 antibody and Akt were purchased from Cell Signaling Technology (US), the β-actin antibody was purchased from Sigma-Aldrich ) And a secondary antibody conjugated with Horseradish peroxidase was purchased from Santa Cruz Biotechnology (USA), and 2- [N- (7-nitrobenz-2-oxa-1,3 (2-NBDG) was purchased from Invitrogen (US), and Superfect Transfection Reagent was purchased from Qiagen (US) , And the Luciferase essay system was purchased from Promega (US).

The following experimental results were processed using SPSS 9.0 (SPSS Inc., Chicago, IL) program. Mean ± SD. The statistical significance of each sample was tested by one-way ANOVA. The difference between the groups was verified by Duncan's multiple test at P <0.05 level.

Example  One : PPAR - gamma  Transcriptional activity evaluation

(4-methoxyphenyl) benzamide (DMPB) on the transcriptional activity of PPAR-αgamma) in HEK293 cells, which is a specific target of the glutazone-based drug, an antidiabetic agent To investigate the PPAR-a transcriptional activity measurement experiment was performed.

In order to transduce the cells, luciferase reporter vector containing the total DNA expression plasmid, PPAR-response element (PPRE), and β-galactosidase for 1 PPAR-α or RXRα and 18 hours later, Were treated with rosiglitazone or DMPB for 24 hours, respectively. Rosiglitazone was used as a positive control. PPAR-α and RXRα activities were measured with the luciferase assay system and normalized using β-galactosidase activity.

As shown in FIG. 1, PPAR-a transcriptional activity by DMPB increased 3.1-fold at 25 ° C and 3.7-fold at 50 ° C, and it was confirmed that transcription activity was increased in a concentration-dependent manner.

Example  2 - DMPB Muscle cell  Differentiation and blood glucose Absorption capacity  evaluation

2-NBDG test was performed to analyze the effect of 2,5-dimethoxy-N- (4-methoxyphenyl) benzamide (DMPB) on direct intracellular glucose uptake.

C2C12 cells were allowed to grow to 100% confluent in normal medium, and then the medium was replaced with culture medium supplemented with 1% horse serum media and cultured for 6 days Lt; / RTI &gt;

Cells that were completely differentiated into muscle cells were incubated overnight in serum-free, low glucose DMEM medium and incubated with 2-NBDG for 24 hours. EX / EM = 485/535 And the degree of blood glucose absorption was measured. Rosiglitazone was used as a positive control.

As shown in FIG. 2, the intracellular glucose uptake of the differentiated C2C12 muscle cells increased 1.5-fold at 25 μM and 2.2-fold at 50 μM, and DMPB could lower blood glucose by promoting glucose uptake into muscle cells .

Example  3: Evaluation of Protein Expression Quantity

The following experiment was conducted to evaluate the effect of 2,5-dimethoxy-N- (4-methoxyphenyl) benzamide (DMPB) on the activation of AMPK, ACC and AKT in C2Cl2 cells.

CCl2 cells were washed twice with phosphate buffered saline (PBS) and resuspended in lysis buffer (50 mM Tris-HCl pH 7.4, 1% NP-40, 0.25% sodium deoxycholate, 150 mM sodium chloride, 1 mM EDTA , 1 mM phenylmethanesulfonyl fluoride (PMSF), 1 mM sodium orthovanadate, 1 mM sodium fluoride (NaF), 1 / / mL aprotinin, 1 / / mL leupeptin and 1 / / mL pepstatin) &Lt; / RTI &gt;

The supernatant was then taken by placing on ice for 5 minutes and then centrifuging at 14,000 rpm for 5 minutes. Sodium dodecyl sulfate (SDS) sample buffer was added to the obtained supernatant, followed by boiling at 100 ° C for 5 minutes, and proteins were separated using 10% SDS PAGE. The separated proteins were transferred to a nitrocellulose membrane and Western blotting analysis was performed using each antibody.

As a result, it was confirmed that DMPB increases phosphorylation of AMPK in C2Cl2 cells in a concentration-dependent manner, and ACC, which is a sub-mechanism of AMPK activation, is also activated by DMPB and phosphorylation of Akt, an insulin-dependent signaling molecule, is also increased 5).

In the above Examples 1 to 3, the N-phenylbenzamide derivatives according to the present invention exhibited a high PPAR-alpha transcriptional activity, a high glucose uptake effect in muscle cells (C2C12), and an AMPK, ACC And AKT, and the pharmaceutical composition and health functional food according to the present invention proved to have an excellent antidiabetic effect.

Claims (6)

delete A pharmaceutical composition for the prophylaxis or treatment of diabetes or diabetic complications comprising a compound of the formula (2)
(2)

Wherein the substituents R ₄ , R ₅ and R ₆ are independently selected from alkoxy groups having 1 to 3 carbon atoms.
A pharmaceutical composition for preventing or treating diabetes mellitus or diabetic complications comprising 2,5-dimethoxy-N- (4-methoxyphenyl) benzamide of the formula (3)
(3)

.
delete A health functional food for preventing or ameliorating diabetes or diabetic complications comprising a compound of the following formula (2) or a salt thereof:
(2)

Wherein the substituents R ₄ , R ₅ and R ₆ are independently selected from alkoxy groups having 1 to 3 carbon atoms.
A health functional food for preventing or ameliorating diabetes or diabetic complications comprising 2,5-dimethoxy-N- (4-methoxyphenyl) benzamide of the following formula (3)
(3)

.

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