KR100907860B1 - Pharmaceutical composition for reducing side effects and increasing therapeutic effect of PPAR-γ agonists - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for reducing side effects of a TZD (thiazolidinedione) -based drug, which is a PPAR-γ agonist. More specifically, the present invention relates to a pharmaceutical composition, including taxa, red antler, ghost, baekrye and broiler, to reduce the side effects and increase the therapeutic effect of the PPAR-γ agonist TZD (thiazolidinedione) drug.

The pharmaceutical composition according to the present invention reduces the side effects of TZD (thiazolidinedione) -based drugs, such as weight gain, weight gain of fat to fat and metabolic dyslipidemia, and improves insulin resistance to reduce the side effects of PPAR-γ agonists. There is an effect that can increase the therapeutic effect.

Description

Pharmaceutical composition for reduction of side effect and improvement of therapeutic effect of PPAR-gamma agonist}

Figure 1 shows the size (A) and body weight change (B) of visceral fat of ICR mice during the 10-week sample administration period.

Figure 2 is a photograph of the morphology of the adipose tissue of ICR mice after 10 weeks of sample administration observed with an optical microscope.

Figure 3 is a graph showing the size of the adipose cells of ICR mice after the administration of the sample for 10 weeks in the number of cells for each diameter.

Figure 4 shows the results of measuring the PPAR-γ gene and SREBP1a gene expression in the adipose tissue of ICR mice after 10 weeks of sample administration by RT-PCR.

The present invention relates to a pharmaceutical composition for reducing side effects of a PPAR-γ agonist. More specifically, the present invention relates to pharmaceutical compositions, including taxa, red antler, ghost, baekrye and broiler, which alleviate the side effects of PPAR-γ agonists and increase the therapeutic effect.

The obesity population, a phenomenon in which fat accumulates in the body due to the lack of exercise and eating habits such as high-fat and high-carbohydrate diets, is rapidly increasing worldwide. Obesity, in particular, is impaired in energy coordination, resulting in metabolic syndrome associated with diseases such as insulin resistance, hypertension, hyperlipidemia, fatty liver and arteriosclerosis, through fat accumulation in the abdominal cavity, leading to type 2 diabetes, especially insulin resistance. Drugs currently available for the treatment of type 2 diabetes are limited to carbohydrate digestion, acarbose (α-glucosidase) inhibitor, and sulfonylureas that promote insulin secretion. sulfonylurea), metformin, which reduces glucose production in the liver, and thiazolidinedione (TZD) -based drugs that increase insulin action as PPAR-γ agonists.

Peroxisome proliferator-activated receptors (PPARs) act as transcription factors as nuclear receptors. There are three isomers (α, δ / β, γ), especially PPAR-γ, which is mainly present in adipocytes, and TZD-based drugs, which are used to treat diabetes, are used as agonists to regulate blood glucose and lipid homeostasis and to control insulin action. Increase.

TZD-based drugs include rosiglitazone, pioglitazone and troglitazone. However, while the TZD-based drugs have an effect of increasing insulin action, side effects of increasing fat weight by inducing adipocyte differentiation have been reported. Furthermore, the administration of PPAR-γ agonists in animals and humans has reported side effects of cardiac hypertrophy, edema, hematotoxicity and hepatotoxicity.

In order to overcome the disadvantages of the conventional PPAR-γ agonist as described above, first, GW007213, N- (9-fluorenyl) methoxycarbonyl (N- (9-fluorenyl) methoxycarbonyl) (FMOC) -L-leucine , PAT5A, and nTZDpa Attempts to develop partial agonists such as, etc., secondly PPAR-γ inhibitors such as berberine, thirdly PPAR-α and γ dual agonists with lipolytic activity. However, these agents have the disadvantage of reducing the efficacy of the PPAR-γ agonist.

The inventors of the present invention, while maintaining the activity of the PPAR-γ agonist, while researching to develop a drug that can improve its side effects, pharmaceutical compositions including taxa, red ox, ghost, baekrye and broiler PPAR- The present invention was completed by confirming that the therapeutic activity can be increased while effectively alleviating the side effects of the γ agonist.

It is therefore an object of the present invention to provide a pharmaceutical composition that alleviates the side effects of PPAR-γ agonists and increases the therapeutic effect.

In order to achieve the object of the present invention as described above, the present invention provides a pharmaceutical composition to reduce the side effects of the PPAR-γ agonist and increase the therapeutic effect.

The pharmaceutical composition according to the present invention includes taxa, red ox, spirit, white and broiler meat, and alleviates the side effects of PPAR-γ agonists and enhances the therapeutic effect.

It is preferable that the said composition is mix | blended in the weight ratio of 5: 3: 3: 3: 1.

The increase in the therapeutic effect may be characterized as being obtained by improving insulin resistance.

The pharmaceutical composition according to the present invention may be composed of a formulation selected from the group consisting of powders, granules, tablets, pills, dragees, capsules, solutions, gels, slurries and suspensions.

Hereinafter, the present invention will be described in detail.

The pharmaceutical composition of the present invention is characterized in that it comprises tack, red ox, ghost, baekrye and broiler. Preferably, the pharmaceutical composition of the present invention is characterized in that the talc, red ox, spirit, white and broiler is formulated in a weight ratio of 5: 3: 3: 3: 1.

The taxa is a tuber of a perennial talus, Alisma orientale, or other cognate root plant belonging to the Alismataceae. Proteins, starches, volatile essential oils, amphiphiles, alkaloids and L-aspargin, phytosterol, β-sitosterol, fatty acids, choline, lecithin, alisol A, B (Alisol A, B) and its acetate ester, Epi-alsiol A. The pharmacological activity of the taxi is known to have edema suppression, diuretic action, blood cholesterol and blood glucose reduction action due to acute and chronic nephritis and hepatic disease, and inhibitory action against Staphylococcus aureus, pneumococcal pneumoniae and tuberculosis.

, Hoelen rubra)이라 한다. 상기 적복령의 약리효과로는 심, 비, 방광경에 들어가 습열을 제거하고 소변의 혼탁(淋濁), 사리(瀉痢) 등을 치료하는 것으로 알려져 있다.The worm is a kind of fungi belonging to the genus Polygonaceae. The peeled outer shell of the ghost is called the ghost of the ghost, and the red part below the surface

Hoelen rubra). The pharmacological effect of the red bokyongyeong is known to enter the heart, rain, bladder, to remove the moist heat and to urinate turbidity, sari (사) and the like.

, Grifola umbellata)은 단풍나무 작목, 떡갈나무, 기타 수목 등의 뿌리에 기생하는 용류로서, 진균부담자낭균문, 진담자낭균아문, 모균아다공과에 속한다. 그리고 상기 저령은 단백질, 가용성당분, 무결정 다당류 에르고스테 린, 조섬유, 무기물을 함유한다. 상기 저령은 해열, 소염, 지갈의 효과가 있는 이뇨제로 주로 종창(腫脹), 복만(腹滿), 열갈(熱渴), 오뇌(懊惱), 열병(熱病), 소변불리(小便不利), 적색열통(赤色熱痛)을 치료한다. 특히 복부내의 열로 인하여 수분이 정체하고, 흡수 배설이 원활하지 못할 때 사용한다. 저령의 분석학적 약리작용에서는 항종양 효과가 규명된 바 있다.Above age (

, Grifola umbellata) is a parasitic parasitic root of maple tree, oak tree, and other trees. And the low contains protein, soluble sugars, amorphous polysaccharides ergosterine, crude fiber, minerals. The age group is a diuretic that has the effect of fever, anti-inflammatory, and jigal, mainly swelling, abdominal pain, fever, blood brain, fever, urinary deficiency, and redness. Treat fever (통 色 熱 痛). In particular, it is used when water is stagnant due to heat in the abdomen, and absorption excretion is not smooth. Antitumor effects have been identified in aging analytical pharmacology.

The baekchul (白 朮, Atractylodis Rhizoma alba) refers to the peeled root of shovel, a perennial herb belonging to the Asteraceae. Its ingredients mainly contain atactic roots of atractone (atractylon), atacticol (atractylol) and vitamins A and D. The pharmacological action of the baekchul is known to hold fat, regulate the humidity (燥濕) and neutralize the action.

The broiler (Cinnamomi Cortex Spissus) is the name of the Chinese medicine of dried cinnamon bark of evergreen trees belonging to the camphor family and cassia. The broiler is widely used in the symptoms of the cold feet and hands that basically appear. As a result of pharmacological experiments, the broiler was found to have a healthy, antibacterial and blood circulation action, and the components include 75% to 90% of cinnamic aldehyde (Cinnamic aldehyder), in addition to cinnamyl acetate, phenylpropyl acetate ( Phenylpropyl acetate).

In the present invention, the herbal medicines can be collected from nature or used commercially. In one embodiment of the present invention was used to purchase each medicine in the domestic Gyeongdong market (Dongdaemun-gu, Seoul).

In addition, the pharmaceutical composition of the present invention may include a mixture of the extracts after mixing the above-described herbal medicines, or after extracting the herbal medicines according to the physical and chemical properties of the active ingredients, respectively.

As the extraction method, it is preferable to use a solvent extraction method known in the art. For example, it may be extracted using any one selected from alcohols such as water, ethanol and methanol, organic solvents such as acetone, ethyl acetate, n-nucleic acid, methanol, diethyl ether, acetone, benzene, or a mixed solvent thereof. have. By the solvent extraction method In preparing the extract, hot water extraction, ultrasonic extraction and reflux extraction methods can be used. In an embodiment of the present invention, the herbal medicines were mixed at a constant ratio, followed by administration of purified water, followed by heat extraction for a predetermined time, followed by filtration to prepare hot water extracts, which were freeze-dried and used in an experiment (see Example 1).

On the other hand, the pharmaceutical composition of the present invention may be formulated in a conventional manner by including the above-described herbal medicine alone or further including one or more pharmaceutically acceptable carriers, excipients or diluents. As used herein, "pharmaceutically acceptable" refers to a composition that is physiologically acceptable and does not cause an allergic reaction, such as gastrointestinal disorders, dizziness, or the like, when administered to a human.

In addition, the pharmaceutical compositions of the present invention formulated as above may be administered via a suitable route of administration. Suitable routes of administration may include several routes including oral, eye drop, transdermal, subcutaneous, intravenous or intramuscular.

The pharmaceutical composition of the present invention may be formulated into a preparation for oral or parenteral administration depending on the route of administration chosen.

Preferably it may be formulated using methods known in the art in the form of preparations for oral administration such as powders, granules, tablets, pills, dragees, capsules, solutions, gels, slurries, suspensions and the like. For example, oral formulations can obtain tablets or dragees by combining the active ingredients with solid excipients and then grinding them, adding suitable auxiliaries and processing them into granule mixtures. Examples of suitable excipients include sugars including lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol and maltitol and starch, cellulose, including starch, corn starch, wheat starch, rice starch and potato starch, etc. Fillers such as cellulose, gelatin, polyvinylpyrrolidone, and the like, including methyl cellulose, sodium carboxymethylcellulose, hydroxypropylmethyl-cellulose, and the like. In addition, crosslinked polyvinylpyrrolidone, agar, alginic acid or sodium alginate and the like may optionally be added as a disintegrant. Furthermore, the pharmaceutical composition may further include an anticoagulant, a lubricant, a humectant, a perfume, an emulsifier, and a preservative.

The dosage of the pharmaceutical composition according to the present invention may be appropriately selected depending on various factors such as the route of administration, the age, sex, weight of the patient and the severity of the patient, and preferably about 5 mg / kg body weight / day-50 mg / Body weight kg / day.

On the other hand, the pharmaceutical composition of the present invention can be administered in combination with the PPAR-γ agonist can reduce the side effects of the PPAR-γ agonist.

An example of the PPAR-γ agonist is a TZD-based drug, and more specifically, refers to pioglitazone.

In addition, examples of the side effects include at least one of weight gain, weight gain of fat to weight and edema.

In one embodiment of the present invention, to determine whether the pharmaceutical composition of the present invention has an effect of reducing the side effects of pioglitazone, a kind of PPAR-γ agonist, ICR mice are fed a normal dietary intake group (RD), a high-fat diet Group (HFD), high-fat diet and 30 mg / kg pioglitazone administered group (HFD + PIO), high-fat diet and 300 mg / kg herbal medicine administration group of Example 1 (HFD + OR), high-fat diet, 30 The pioglitazone of mg / kg and the 300 mg / kg administration group of the herbal medicine of Example 1 (HFD + PIO + OR) were divided and the change in body weight and organ weight was measured for 10 weeks (see Example 3). As a result, it was found that the pharmaceutical composition according to the present invention has an effect of reducing visceral fat and body weight increased by administration of pioglitazone (see FIG. 1 and Table 2).

In another embodiment of the present invention, the result of observing the change in the adipose tissue of the ICR mouse (see Example 4), it can be seen that the adipose tissue enlarged by pioglitazone is reduced due to the administration of the pharmaceutical composition according to the present invention (See FIG. 2), the diameter distribution of adipocytes was also reduced due to the administration of the pharmaceutical composition according to the present invention (see FIG. 3).

In another embodiment of the present invention, the change in blood lipid concentration of the ICR mouse was measured (see Example 6). As a result, it was found that the blood triglyceride, total cholesterol and LDL-cholesterol increased by pioglitazone were inhibited by administration of the pharmaceutical composition of the present invention (see Table 4).

Furthermore, in another embodiment of the present invention, the expression change of PPAR-γ gene involved in glucose uptake and SREBP1a gene involved in fat synthesis in adipose tissue of the ICR mouse was measured by RT-PCR. See example 7). As a result, it was shown that the expression of SREBP1a and the expression of PPAR-γ increased by pioglitazone are reduced by administration of the herbal medicine according to the present invention (see FIG. 4). From this, it could be judged that the pharmaceutical composition of the present invention inhibits differentiation into adipocytes by pioglitazone.

As described above, the pharmaceutical composition of the present invention was found to have an activity of reducing side effects of pioglitazone such as weight gain, weight gain of fat to fat, and lipid metabolism.

In addition, the pharmaceutical compositions of the present invention may enhance the therapeutic effect of PPAR-γ agonists. Preferably, the pharmaceutical composition of the present invention may enhance the therapeutic effect of the PPAR-γ agonist by improving the insulin resistance of the PPAR-γ agonist.

In one embodiment of the present invention, the improvement effect on the blood glucose, insulin and insulin resistance indicators of the ICR mouse was investigated (see Example 5). As a result, co-administration of pioglitazone and the pharmaceutical composition according to the present invention was confirmed to have a synergistic effect in improving insulin resistance (see Table 3).

Hereinafter, the specific method of the present invention will be described in detail with reference to Examples, but the scope of the present invention is not limited only to these Examples.

< Example  1>

Preparation of the pharmaceutical composition according to the invention

In the prescription composition of the pharmaceutical composition according to the present invention, each herbal medicine was purchased in Kyungdong market and mixed in the ratio shown in Table 1, more specifically, 10 g of Taxa, 6 g of Red Riding Age, 6 g of Age, 6 g of Baekchul, Broiler 7 g of mixed herbal medicine consisting of 2 g (210 g) and about 1300 ml of purified water were placed in a medicine bath, heated for 2 hours, filtered through a filter paper, concentrated under reduced pressure, and lyophilized.

Name and content of herbal medicine contained in the pharmaceutical composition of the present invention Chinese medicine name Herbal medicine content Taxi Alismatis rhizoma 10 g

)Red Riding Age

) Hoelen rubra 6 g Age

) Grifola umbellata 6 g Baekchul (白 朮) Atractylodis Rhizoma alba 6 g Broiler Cinnamomi Cortex Spissus 2 g Total content per pack 30 g

< Example  2>

Breeding and Experimental Design of Laboratory Animals

Four-week-old male ICR mice were purchased from Orient Co., Ltd. and used. Rats are kept in a plastic mouse cage and kept in an animal room under conditions of 23 ± 2 ° C, 50 ± 10% relative humidity, and light / dark cycles (12/12 hr), water and feed (LabDiet). Should be taken freely. Animals were acclimated to the environment for one week prior to the start of the experiment and divided into five groups as follows: Drugs were administered orally: normal diet (RD), high-fat diet (HFD), high-fat diet and 30 mg. pioglitazone-administered group (HFD + PIO), high-fat diet and 300 mg / kg pharmaceutical composition-administered group (HFD + OR), high-fat diet, 30 mg / kg pioglitazone and Example 1 The pharmaceutical composition of 300 mg / kg administration group (HFD + PIO + OR) was divided into experiments for 10 weeks. Body weight was measured weekly and dietary and negative amounts were measured every two days. After 10 weeks, the weight of liver and adipose tissue was measured in each group.

<Example 3>

Changes in Body Weight and Organ Weight of Laboratory Animals

Body weights were measured during the 10-week sample administration period, and mice were opened at the end of the experiment to compare the size of visceral fat with the naked eye. In addition, the liver tissue of the mouse and the adipose tissue of the epididymis were extracted, and each weight was measured. The measured results were tested by Student's t-test and the significance of the mean difference between the groups was above p <0.05 and the results were shown along with the standard error.

As a result, the weight of the last day was in the order of high-fat diet group (HFD)> HFD + pioglitazone administration group (PIO)> HFD + PIO + pharmaceutical composition administration group (OR)> HFD + OR (Example 1B) . HFD + PIO increased visceral fat size compared to HFD group, while HFD + PIO + OR decreased visceral fat size compared to HFD + PIO. Was observed (FIG. 1A).

After 10 weeks of sample administration, body weight and organ weights were compared between the groups, indicating that the HFD + OR-administered group lost weight (14.7%) compared to the HFD group. In addition, it can be seen that the weight gain of the HFD + PIO + OR administration group compared with the HFD + PIO administration group decreased (6.6%). In the HFD + PIO group, the weight of fat was increased (34%) compared to the HFD control group, whereas the HFD + PIO + OR group was suppressed (29%). Can be. From this, it can be seen that the pharmaceutical composition according to the present invention has an effect of suppressing weight gain due to high-fat diet ingestion and administration of pioglitazone (Table 2).

Changes in Body Weight and Organ Weight Following Administration of the Pharmaceutical Compositions of the Invention group Body weight (g) Start of experiment Experiment end point Weight gained Liver / weight (%) Fat / weight (%)  RD 27.8 ± 1.3 33.7 ± 1.4 6.0 4.4 2.7  HFD 28.8 ± 1.0 42.4 ± 1.8 13.6 3.9 4.4  HFD + PIO 27.8 ± 0.6 41.5 ± 1.1 13.7 3.5 5.9  HFD + OR 28.1 ± 1.0 39.7 ± 2.3 11.6 3.7 4.1  HFD + PIO + OR 27.8 ± 0.5 40.6 ± 2.0 12.8 3.8 4.2

<Example 4>

Effect on adipose tissue

After 10 weeks of sample administration, the adipose tissue extracted from each mouse was fixed using 10% neutral buffered formalin. After the process of dehydration and embedding, a paraffin block was made and made into tubular sections having a thickness of 5 μm. Then, paraffin was removed with xylene, and 100%, 95%, 90%, 80%, 70% alcohol was used. Hydrated. The staining method was dehydrated by Hematoxylin-Eosin staining, sealed with Canadian balsam, and observed under an optical microscope (Olympus, Japan).

As a result, in the HFD + OR administration group, the size of the adipose tissue was significantly reduced compared to the HFD control group, it was confirmed that the adipose tissue of HFD + PIO + OR also significantly reduced compared to the HFD + PIO administration group (Fig. 2). It is believed that the adipose tissue enlarged by pioglitazone is reduced due to the administration of the pharmaceutical composition according to the present invention.

In addition, as a result of showing the adipocyte size as the number of cells for each diameter, the diameter distribution of the adipocytes showed that the HFD + OR and the HFD + PIO + OR-administered groups reduced the number of large-diameter cells compared to the HFD control group (FIG. 3). ).

<Example 5>

Improvement effects on blood glucose, insulin and insulin resistance indicators

After 10 weeks of sample administration, each mouse was fasted for 12 hours, and blood was collected. Whole blood obtained through the orbital vein was centrifuged at 5,000 rpm for 5 minutes and then plasma was used for analysis. Blood glucose concentration was measured by glucose oxidase method (Trinder method) and absorbance was measured by UV Spectrophotometer (UV-3, U-3210, HITACHI; Japan). Blood insulin concentration was measured with an ELISA reader (Labsystems, Finland) by purchasing an insulin insulin ELISA kit (insulin ELISA kit, Shibayagi, Japan). Insulin resistance index was calculated according to the following formula, and the measured results were statistically treated in the same manner as in Example 3.

Insulin Resistance Index (HOMA-IR) = Fasting Insulin Blood (μU / ml) x Fasting Blood Glucose Level (mM) /22.5

As a result, the HFD + PIO group was significantly lower in blood glucose than the HFD control group (36%). Interestingly, the blood insulin level was 58% decreased in the group administered with the pharmaceutical composition according to the present invention compared to the HFD control group, and 85% compared with the HFD control group in the group treated with pioglitazone and the pharmaceutical composition according to the present invention, HFD + PIO. It was 41% lower than the group. Although the pharmaceutical composition of the present invention had little effect on reducing blood glucose, the HDF + PIO + OR group showed a 43% reduction in the insulin resistance index as a result of a significant decrease in blood insulin levels. (Table 3). Therefore, the combination of pioglitazone and the pharmaceutical composition according to the present invention is expected to have a synergistic effect in improving insulin resistance.

Changes in Blood Glucose, Insulin, and Insulin Resistance Indicators group Glucose concentration (mM) Insulin (mU / ml) HOMA-IR Start of experiment Experiment end point  RD 4.9 ± 0.4 5.0 ± 0.4 29.56 ± 11.86 6.57  HFD 4.9 ± 0.2 7.8 ± 0.5 207.27 ^** ± 61.22 71.85  HFD + PIO 4.8 ± 0.3 5.8 ± 0.4 53.29 ^** ± 22.7 13.7  HFD + OR 4.9 ± 0.4 7.7 ± 0.8 86.91 ^* ± 24.76 29.74  HFD + PIO + OR 4.9 ± 0.2 5.6 ± 0.4 31.57 ^** ± 4.80 7.86

*: There is a significant difference at p <0.05.

**: There is a significant difference at p <0.01.

<Example 6>

Changes in blood lipid concentration

After 10 weeks of sample administration, each mouse was fasted for 12 hours, and blood was collected. Whole blood obtained through the orbital vein was centrifuged at 5,000 rpm for 5 minutes and then plasma was used for analysis. Blood triglycerides, total cholesterol, HDL cholesterol, and LDL cholesterol were measured by purchasing each kit from Yeongdong Pharm. The measured results were statistically treated in the same manner as in Example 3.

As a result, HFD + PIO group showed 25%, 18% and 88% increase in triglyceride (TG), total cholesterol (TC) and LDL-C levels, respectively, compared to HFD control group. However, the HFD + OR group increased 8%, 23%, and 35% in TG, TC, and HDL-C levels, respectively, and decreased 37% in LDL-C compared to the HFD control group. The HFD + PIO + OR group showed 1.7%, 8.9%, and 42.4% reductions in TG, TC, and LDL-C levels, respectively, compared to the HFD + PIO group (Table 4). These results indicate that blood triglycerides, total cholesterol and LDL-cholesterol increased by pioglitazone are inhibited by the pharmaceutical composition of the present invention.

Blood lipid concentration group Triglycerides (mg / dl) Total cholesterol (mg / dl) HDL-cholesterol (mg / dl) LDL-cholesterol (mg / dl)  RD 73.8 ± 11.2 125.8 ± 5.4 93.8 ± 9.1 17.2 ± 1.5  HFD 90.6 ± 5.6 140.9 ± 8.5 101.1 ± 10.7 34.6 ± 11.7  HFD + PIO 113.4 ± 6.8 166.5 ± 15.7 127.3 ± 5.6 ^* 65.5 ± 12.0  HFD + OR 98.3 ± 6.9 165.1 ± 12.9 136.3 ± 7.9 ^** 21.9 ± 3.0  HFD + PIO + OR 111.5 ± 11.7 ^* 151.6 ± 14.9 105.0 ± 2.4 37.7 ± 13.4

*: There is a significant difference at p <0.05.

**: There is a significant difference at p <0.01.

<Example 7>

Effect of SREBP1a and PPAR-γ Gene Expression

The effects on the expression of PPAR-γ gene involved in glucose uptake and SREBP1a gene involved in fat synthesis in adipose tissue of each mouse were analyzed using RT-PCR.

To this end, total RNA was first isolated from the adipose tissue of mice using the guanidine thiocyanate-water saturated phenol / chroloform method (Chomczynski and Sacchi, 1987). Total RNA in the water layer was precipitated using isopropanol, and the isolated RNA was quantified by measuring absorbance at wavelengths of 260 nm and 280 nm.

RT-PCR was performed by reverse transcription of 1 μg of total RNA using MMLV reverse transcriptase (Moloney murine leukemia virus transcriptase) and primers of Table 5 to prepare cDNA, and then PCR amplification using the template as a template. Primer was prepared in 25 μl reaction solution containing 20 mM Tris-HCl (pH 8.4), 50 mM KCl, 1.5 mM MgCl ₂ , 0.5 mM dNTP, 5 μl cDNA and 2.5 units of Taq DNA polymerase. mM was added, and PCR reaction conditions were repeated 30 times by denaturing at 95 ° C. for 1 minute, attaching at 55 ° C. for 1 minute, and extending at 72 ° C. for 2 minutes. PCR amplification products were electrophoresed at 100V using a 1% agarose gel containing 0.5 μg / ml ethidium bromide, and β-actin was used as a control of the amplified genes.

Primer sequence used for RT-PCR primer order SEQ ID NO: PPAR-γ sense AGG CCG AGA AGC TGT TG One PPAR-γ antisense TGG CCA CCT CTT TGC TCT GCT C 2 SREBP1a sense GCG CYA CCG GTC TTC TAT CA 3 SREBP1a antisense TGC TGC CAA AAG ACA AGG G 4 β-actin sense GTC GTA CCA CTG GCA TTG TG 5 β-actin antisense GCC ATC TCC TGC TCA AAG TC 6

As a result, the expression of SREBP1a was significantly increased in the HFD + PIO group compared with the HFD control group, and the expression of PPAR-γ was also increased in the HFD + PIO group compared to the HFD control group. On the other hand, the HFD + PIO + OR-administered group showed decreased gene expression of PPAR-γ and SREBP1a compared to the HFD + PIO-administered group (FIG. 4).

From this, it is believed that the pharmaceutical composition of the present invention inhibits differentiation into adipocytes by pioglitazone. Therefore, the pharmaceutical composition of the present invention was found to have the activity of reducing the side effects of pioglitazone.

As described above, the pharmaceutical composition according to the present invention reduces the side effects of PPAR-γ agonists such as weight gain, weight gain of fat and lipid metabolism and improves insulin resistance to treat PPAR-γ agonists. There is an effect that can increase the effect. Therefore, the co-administration of the pharmaceutical composition of the present invention and the PPAR-γ agonist is very useful for the prevention and treatment of diabetes.

<110> Industry Academic Cooperation Foundation of KyungHee University <120> Pharmaceutical composition for reduction of side effect of          PPAR-gamma agonist <160> 6 <170> KopatentIn 1.71 <210> 1 <211> 17 <212> DNA <213> Artificial Sequence <220> <223> PPAR-gamma sense primer <400> 1 aggccgagaa gctgttg 17 <210> 2 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> PPAR-gamma antisense primer <400> 2 tggccacctc tttgctctgc tc 22 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> SREBP1a sense primer <400> 3 gcgcyaccgg tcttctatca 20 <210> 4 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> SREBP1a antisense primer <400> 4 tgctgccaaa agacaaggg 19 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> beta-actin sense primer <400> 5 gtcgtaccac tggcattgtg 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> beta-actin antisene primer <400> 6 gccatctcct gctcaaagtc 20  

Claims (7)

Weight loss, weight gain and fat to fat caused by side effects of TZD (thiazolidinedione) -based drug, including PPAR-γ agonist And pharmaceutical composition for alleviating at least one of lipometabolic disorders. delete The composition of claim 1, wherein the TZD-based drug is pioglitazone. It is a pharmaceutical agent that increases the diabetic treatment effect of TZD (thiazolidinedione) drug, which is a PPAR-γ agonist, by including a weight ratio of 5: 3: 3: 3: Composition. delete The composition of claim 4, wherein the TZD-based drug is pioglitazone. 7. A compound according to any one of claims 1, 3, 4 and 6, selected from the group consisting of powders, granules, tablets, pills, dragees, capsules, solutions, gels, slurries and suspensions. A composition consisting of formulations.

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