KR101349187B1 - Pharmaceutical composition for treating and preventing lipid metabolism-related diseases - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the treatment and prevention of fat metabolic disorders, the pharmaceutical composition of the present invention comprising a compound of formula 1 as an active ingredient isosilicate dehydrogen that plays an important role in in vivo energy metabolism in human hepatocytes Increases the activity of the enzyme 3α (IDH3α) promoter and the carnitine palmitoyl transferase 1A (CPT1A) promoter, leading to a decrease in blood sugar and triglycerides and consequently a weight loss, resulting in fat metabolic disorders such as obesity, hyperlipidemia and diabetes It can be usefully used for the prevention and treatment of diseases.

Description

Pharmaceutical composition for the treatment and prevention of fat metabolic disorders {PHARMACEUTICAL COMPOSITION FOR TREATING AND PREVENTING LIPID METABOLISM-RELATED DISEASES}

1 is a graph of luciferase activity observed and treated with a compound of the present invention for cells containing an IDH3α promoter-luciferase fusion construct in Example 1. FIG.

FIG. 2 is a graph of luciferase activity observed and treated with a compound of the present invention for cells containing the CPT1A promoter-luciferase fusion construct in Example 1. FIG.

3 is a graph showing the left / right triglyceride weight ratio of the compound of the present invention in Example 2 was applied to only the left side of the back of the diabetic mouse and observed.

Figure 4 is a graph of blood glucose change observed and administered to the compound of the present invention in obese mice in Example 3.

The present invention relates to a pharmaceutical composition useful for the treatment and prevention of fat metabolism abnormal diseases such as obesity, hyperlipidemia, diabetes.

The process of breaking down lipids and using them as energy sources can be divided into three stages. In the first step, they are decomposed into constituents such as fatty acids, and in the second step, the decomposed components become acetyl-CoA through β-oxidation. It is completely oxidized in the TCA cycle (tricarboxylic acid cycle). Various energy sources are metabolized to acetylcoa, which is oxidized to carbon dioxide (CO ₂ ) and water (H ₂ O) (TCA cycle), all of which are enzymes present in the matrix or inner membrane of mitochondria. Caused by the military In the TCA cycle, citrate is shifted to the position of the hydroxyl group (-OH) by an aconitase to become an isocitrate, which isocitrate dehydrogenase. Oxidized by 3α (IDH3α) to form oxalosuccinate, an intermediate in the state of binding to the enzyme, and reducing the coenzyme NAD ⁺ and decarboxylated immediately to produce alpha-ketoglutarate and carbon dioxide ( CO ₂ ). The generated NADH is immediately transferred to the electron transport system present in the mitochondrial substrate to generate ATP, which is an active energy in vivo. Therefore, increasing the activity of IDH3α increases the activity of the TCA circuit, which can finally oxidize and degrade lipids to produce ATP energy ( Journal of Biological Chemistry , 22199-22205 (1991)). In the event of problems with the TCA circuit, ATP synthesis, which is an active energy in vivo, is reduced, fatty acid synthesis is increased, and β-oxidation is also reduced, resulting in obesity ( Journal of Theoretical Biology , 33-). 44 (2003)]. Eventually, it is expected that substances that modulate the expression of IDH3α may help to treat obesity.

Meanwhile, the carnitine palmitoyl transferase 1A (CPT1A) enzyme is a protein present in the mitochondria in cells and serves to transfer long chain fatty acids into the mitochondria. This enzyme is known to be involved in the rate determining step in the oxidation of long chain fatty acids in the intracellular mitochondria. Increasing the expression of CPT1A in cell-level and animal-level experiments promotes oxidation of fatty acids resulting in increased energy consumption. Hyperlipidemia induced by high-fat diet was induced in Zucker rats that induced obesity through high-fat diet administration and treated with tetratradecylthioacetic acid, a drug that promotes fatty acid beta oxidation through increased expression of CPT1A. Improved and increased insulin sensitivity has been reported ( Progress in Lipid Research , 40 , 231-268 (2001)), and ragaglitazar, which has been shown to increase the activity of CPT1A in the liver, has been reported to have a high fat diet. Treatment with hamsters has been shown to reduce the amount of fat in the body ( British Journal of Pharmacology , 140 , 527-537 (2003)). In light of these results, it is expected that substances that increase the expression of CPT1A may contribute to the treatment and prevention of obesity and hyperlipidemia by promoting the oxidation of fatty acids in cells, increasing energy consumption and reducing the amount of fat in the body. .

Accordingly, it is an object of the present invention to provide a pharmaceutical composition useful for the treatment and prevention of fat metabolic disorders such as obesity, hyperlipidemia and diabetes.

In order to achieve the above object, the present invention provides a pharmaceutical composition for the treatment or prevention of fat metabolic disorders, comprising the compound of formula 1 as an active ingredient:

Where

R ₁ is H or C ₁ -C ₅ alkyl,

R ₂ is H or OH,

n is an integer of 1 to 3;

Hereinafter, the present invention will be described in more detail.

The compound of Formula 1 may be easily prepared according to the method disclosed in International Patent Application No. PCT / KR2006 / 001006, and a commercially available one may be purchased and used if necessary.

The compound of Formula 1 increases the activity of the promoters of isocitrate dehydrogenase 3α (IDH3α) and carnitine palmitoyl transferase 1A (CPT1A) among proteins involved in energy metabolism in vivo. It leads to a decrease in blood sugar and a decrease in triglycerides in subcutaneous fat tissue, and consequently to a weight loss, effectively preventing and treating fat metabolism abnormal diseases such as obesity, hyperlipidemia and diabetes.

The compound of Formula 1 may be mixed with a suitable pharmaceutically acceptable carrier or excipient or diluted with a diluent according to a conventional method to prepare a pharmaceutical composition having the above function. Examples of suitable carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. The pharmaceutical composition may further include fillers, anti-coagulants, lubricants, wetting agents, flavors, emulsifiers, preservatives, disintegrants, sweeteners, lubricants, flavoring agents and the like.

The pharmaceutical compositions of the invention may be formulated using methods well known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal. The formulations may be in the form of tablets, pills, powders, sachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols, soft or hard gelatin capsules, sterile injectable solutions, sterile powders and the like.

The pharmaceutical compositions of the invention can be administered via several routes including oral, transdermal, subcutaneous, intravenous or intramuscular. Typical daily dosages of the active compounds of the present invention range from 0.1 to 100 mg / kg body weight and may be administered once or in divided doses. However, it is to be understood that the actual dosage of the active ingredient should be determined in light of several relevant factors such as the route of administration, the age, sex and weight of the patient, and the severity of the disease, and therefore the dosage should in any way be within the scope of the invention. It is not intended to be limiting.

Hereinafter, the present invention will be described in more detail based on the following examples. However, the following examples are illustrative of the present invention but are not limited thereto.

Manufacturing example

Compounds 1 to 3 having the structure of Chemical Formula 1 were prepared according to the method described in International Patent Application PCT / KR2006 / 001006, and are shown in Table 1 below.

Example 1 Activity Measurement of IDH3α and CPT1A Promoters

(Step 1) Cell Line and Cell Culture

10% blood serum (Dulbecco's modifided Eagle's Medium, Invitrogen, 1210-0038) containing fetal borine serum and 10% penicillin / streptomycin antibiotic (Invitrogen, 15140-122) at 37 ° C., 5% CO Incubated under ₂ conditions.

(Step 2) Determination of activity of each of IDH3α promoter and CPT1A promoter

Each cell line cultured in step 1 was trypsinized to prepare a single cell suspension, and cultured in a 96 well plate containing DMEM medium for 24 hours. Thereafter, the compound of Preparation Example dissolved in DMSO (dimethyl sulfoxide) was added to the cells at a concentration of 10 ppm, and further incubated for 24 hours. Thereafter, the activity of the promoter was measured by measuring the activity of luciferase expressed using a Luciferase assay kit from Promega. A graph of luciferase activity by IDH3α promoter is shown in FIG. 1, and a graph of luciferase activity by CPT1A promoter is shown in FIG. 2, respectively.

1 and 2, it can be seen that the compounds of the present invention promote the activity of the IDH3α promoter and the CPT1A promoter.

Example 2 Isolation and Measurement of Triglycerides from Subcutaneous Fatty Tissue in Diabetic Mice

(Step 1) Compound Treatment and Subcutaneous Subcutaneous Tissue Extraction in Diabetic Mice

After depilation of hair on the back of diabetic mice (db / db mice) (10 weeks old, a central animal experiment), the compound 2 of Preparation Example 2 was added to the delivery solution (polyethylene glycol 400: ethanol = 7: 3) at a concentration of 2%. 130 μl of the dissolved solution was applied to the left half of the back of the diabetic mouse and applied for 5 days, morning and afternoon, until it soaked into the skin. On the right, only gastric delivery solution was applied and used as a control. The diabetic mice treated as above were sacrificed by cervical spinal bone, and then, using surgical scissors, subcutaneous fat tissue was removed and placed on a flat rubber plate. Each of these subcutaneous fat tissues was extracted using a 4 mm diameter biopsy punch (Biopsy punch, Stiefel Co., D6307).

(Step 2) Separation of triglycerides from the extracted subcutaneous fat tissue and analysis by TLC

Subcutaneous fat tissue removed in step 1 was placed in a round flask containing 10 ml of chloroform / methanol (2: 1) solution for 18 hours to dissolve fats (including triglycerides) in the solution. Each of these sample solutions was placed in a 1.5 ml vial for thin layer chromatography (TLC). Meanwhile, TLC plates (20 × 10 cm; Camag Cat No. 034.5642) were prepared by placing them on a TLC plate heating stand (Cammg Cat No. 022.3306) and preheating at 100 ° C. for 10 minutes.

A small bottle containing the TLC plate and the sample solution was mounted in an automatic TLC sampler (Camag Cat No. 022.7400), and then 2 µl of each sample solution was prepared using the WinCATS program (Camag Cat No. 027.6340). Each plate was placed on the plate. Thereafter, the TLC plate loaded with the sample solution was mounted on an automated multiple development device (Automated multiple development, Cammg Cat No. 022.8660), and then each sample solution was developed using the solvents listed in Table 2 in order. At this time, TLC was performed using the AMD2 program (Camag Cat No. 027.6350).

The developed TLC plate was reacted with staining reagent (10% CuSO ₄ , 8% phosphoric acid) for 15 minutes, and then the TLC plate was placed on a heating table and heated at 165 ° C. for 10 minutes. After the reaction was completed, the TLC plate showing triglycerides was scanned on Powerlook 2100 × L (UMAX, Cat No. U9901-H9DO), and then the image master built in the computer was used. The amount of triglycerides from each sample was measured, and the results are shown in the left / right triglyceride weight ratio graph in FIG. 3.

From the graph of Figure 3, it can be seen that the compound of the present invention can reduce the triglyceride by the compound of the present invention, such as bringing a left / right triglyceride weight ratio as low as 5% (p value <0.5).

Example 3 Analysis of Effect on Blood Sugar

For 8 week-old male obese mice (ob / ob mice, Polars International, Korea) genetically induced obesity, freely ingested AIN93G feed (Han Life R & D, Korea) containing 0.02% of compound 2 of the above preparation for 2 weeks. As a control, only AIN93G feed was also freely consumed for 2 weeks. After 2 weeks, blood glucose was measured from blood collected and the results are shown in FIG. 4.

From the graph of Figure 4, it can be seen that the compound of the present invention has a blood sugar reducing effect.

As described above, the pharmaceutical composition of the present invention comprising the compound of formula 1 increases the activity of the IDH3α promoter and the CPT1A promoter in proteins involved in energy metabolism in vivo, thereby reducing blood sugar and triglycerides, obesity, hyperlipidemia It can be usefully used for the prevention and treatment of fat metabolism abnormal diseases such as diabetes.

Claims (7)

A pharmaceutical composition for treating or preventing adipose metabolism abnormal disease, comprising the compound of formula 1 as an active ingredient: Formula 1

Where R ₁ is H or C ₁ -C ₅ alkyl, R ₂ is H or OH, n is an integer of 1 to 3; The method of claim 1, Fatty metabolic disorders are obesity, hyperlipidemia or diabetes, characterized in that the pharmaceutical composition. The method of claim 1, A pharmaceutical composition characterized by promoting the activity of an isolate dehydrogenase 3α (IDH3α) promoter. The method of claim 1, A pharmaceutical composition characterized by promoting the activity of a carnitine palmitoyl transferase 1A (CPT1A) promoter. The method of claim 1, Pharmaceutical composition, characterized in that to reduce blood sugar. The method of claim 1, Pharmaceutical composition, characterized in that to reduce triglycerides. The method of claim 1, Pharmaceutical composition, characterized in that to reduce weight.

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