KR101341200B1 - Pharmaceutical composition for treating or preventing metabolic syndrome comprising ＴＲＡＰ８０ regulator - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for treating or preventing metabolic syndrome, which contains a TRAP80 modulator as an active ingredient, wherein the antibody specifically binding to siRNA or TRAP80 that inhibits TRAP80 expression is selected from SREBP-1c. By selectively inhibiting transcription, metabolic syndromes such as arteriosclerosis and fatty liver can be effectively treated or prevented.
In addition, pharmaceutical compositions containing LXR agonists in combination with TRAP80 modulators can inhibit fatty acid metabolic activities such as serum triglycerides and hepatic triglycerides, which are unwanted effects of LXR agonists, increasing serum HDL and reducing arteriosclerosis. Since the desired effect of the LXR agonist can be enhanced, the effect as a therapeutic agent for metabolic syndrome such as atherosclerotic disease or fatty liver by the LXR agonist can be further improved.

Description

Pharmaceutical composition for treating or preventing metabolic syndrome comprising TrAp500 regulator}

The present invention relates to a pharmaceutical composition for treating or preventing metabolic syndrome by selectively inhibiting transcription of SREBP-1c by containing a TRAP80 modulator as an active ingredient to effectively treat or prevent metabolic syndrome such as atherosclerosis and fatty liver.

Metabolic syndrome refers to a syndrome that is accompanied by risk factors such as obesity, diabetes, hypertriglyceridemia, hypertension, liver disease, cardiovascular disease, and blood clotting abnormalities. The symptom itself is not fatal, but because it has a predisposition to develop serious diseases such as diabetes and ischemic cardiovascular disease, it is a serious threat to modern people.

To prevent atherosclerosis during metabolic syndrome, it is important to lower blood cholesterol levels. A strong cholesterol-lowering effect of statins has been reported in patients with hypercholesterolemia. Statins inhibit hydroxymethylglutaryl-CoA reductase, a rate-determining enzyme for cholesterol biosynthesis, thereby inhibiting cholesterol biosynthesis in vivo and thus lowering cholesterol content in the liver and thus increasing LDL receptors. Lowers cholesterol levels in plasma. However, despite a long time since the development of such drugs, it has been reported that there is no change in the incidence of coronary artery disease. Therefore, inhibition of arteriosclerosis is insufficient only by the reduction of cholesterol.

In addition, this treatment is also important because there is no safe and effective treatment for fatty liver disease, one of the metabolic syndrome.

On the other hand, Liver X receptor (LXR), a kind of nuclear receptor superfamily, is a nuclear cholesterol sensor that regulates the expression of various important proteins related to lipid metabolism and inflammation. LXR is activated by oxysterols such as 22 (R) -hydroxycholesterol, 24 (S) -hydroxycholesterol and 24,25 (S) -epoxycholesterol.

Two LXR proteins α and β were found in mammals, while LXRβ was found in almost all tissues tested, while the expression of LXRα was significantly limited and found at the highest levels in the liver, kidney, small intestine, spleen, and It was found at lower levels in the adrenal glands.

LXRα regulates the expression of genes involved in cholesterol synthesis, its intracellular outflow (Fielding & Fielding, Biochim. Biophys. Acta, 2001, 1533, 175-89) and cholesterol metabolism. Thus, LXRα agonists can be usefully used to regulate blood cholesterol levels by themselves or in combination with known cholesterol-biosynthetic inhibitor drugs, statins. Recent studies on LXR agonists such as T0901317 and hypocollamide have further convinced that cholesterol levels can be regulated by LXR agonists ( Genes & Development , 14 (22): 2831-8, 2000; Steroids , 66: 673-81, 2001).

Increased transcription of ABCA1 by LXR agonists such as T0901317 or GW3965 can contribute to increased serum HDL and reduced atherosclerosis, and this desirable effect of LXR agonists can be utilized as therapeutics in patients with atherosclerotic disease. . However, LXR activation is associated with increased serum triglycerides and hepatic triglyceride contents in association with increased mRNA of SREBP-1c, an important regulator of de novo fatty acid synthesis.

Therefore, there is a need to separate the negative effects of LXR on fatty acid metabolism from the positive effects of LXR on cholesterol metabolism, which can be usefully used for developing metabolic syndrome treatments such as atherosclerosis or fatty liver.

Accordingly, the present inventors identified coactivators that interact with LXRa, and it was confirmed that TRAP80 stimulates SREBP-1c transcription in a ligand-dependent manner through the LXR response element (LXRE). In addition, transcription of SREBP-1c was significantly inhibited by siRNA of TRAP80, and transcription of ABCA1 was not inhibited. These findings have completed the present invention in light of the fact that TRAP80 modulators can be used to treat or prevent metabolic syndrome and can augment the useful effects of LXR agonists.

In order to solve the problems of the prior art, an object of the present invention is to provide a pharmaceutical composition for treating or preventing metabolic syndrome, such as arteriosclerosis or fatty liver containing a TRAP80 modulator as an active ingredient.

In addition, another object of the present invention is to treat or prevent metabolic syndrome, such as arteriosclerosis or fatty liver, including a TRAP80 modulator with an LXR agonist that can inhibit unwanted fatty acid metabolic activity while increasing the useful effect of the LXR agonist. It is to provide a composition.

Another object of the present invention is to provide a method for screening a drug for treating or preventing metabolic syndrome such as arteriosclerosis or fatty liver using a TRAP80 modulator.

Another object of the present invention is to provide a method for diagnosing metabolic syndrome such as atherosclerosis or fatty liver using TRAP80 expression profile.

It is still another object of the present invention to provide a method for reducing side effects of LXR agonists that inhibit the interaction between LXR and TRAP80 by administering a TRAP80 modulator.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating metabolic syndrome, characterized in that it contains a TRAP80 modulator as an active ingredient.

The TRAP80 modulator may be any one selected from the group consisting of siRNA that inhibits TRAP80 expression and an antibody that specifically binds to TRAP80, and may selectively inhibit transcription of SREBP-1c.

SiRNA molecules that inhibit TRAP80 expression used in the present invention may have a form in which a short nucleotide sequence (eg, about 5-15 nt) is inserted between a self-complementary sense and an antisense strand, in which case SiRNA molecules formed by the expression of nucleotide sequences will form a hairpin structure by intramolecular hybridization, and form a stem-and-loop structure as a whole. These stem-and-loop structures are processed in vitro or in vivo to produce active siRNA molecules that can mediate RNAi.

The antibody that specifically binds to TRAP80 used in the present invention is a polyclonal or monoclonal antibody, preferably a monoclonal antibody. Such antibodies may be prepared by methods commonly practiced in the art, such as fusion methods ( European Journal of Immunology , 6: 511-519, 1976), recombinant DNA methods (US Pat. No. 4,816,56) or phage antibody libraries. Method ( Nature , 352: 624-628, 1991; J. Mol. Biol. , 222: 58, 1-597, 1991).

According to the present invention, coactivator proteins interacting with LXRa ligand binding domain (LBD) were identified, and these proteins were identified by immunoprecipitation analysis using Flag-LXRa and in vivo interactions. GST pull-down analysis confirmed TRAP80 and TRAP220 binding directly to LXRa out of five coactivator proteins. TRAP80 interacts GST-LXRa LBD with ligand-independently, but LXRa of both synthetic and natural promoters Mediated transcription was stimulated ligand dependent. TRAP80 recruitment of the SREBP-1c promoter to LXRE was ligand dependent and TRAP80 siRNA strongly inhibited LXR agonist dependent SREBP-1c expression, but did not inhibit ABCA1 expression.

In other words, TRAP80 selectively promotes transcription of the SREBP-1 gene via a ligand-dependent LXR-dependent pathway, and a novel approach that inhibits the interaction of TRAP80-LXR could effectively treat and prevent metabolic syndrome such as atherosclerosis or fatty liver. Can promote the development of new drugs or LXR agonists.

The metabolic syndrome includes arteriosclerosis, liver disease, hyperlipidemia, hypercholesterolemia, obesity, hypertension, hyperinsulinemia, hyperglycemia, type 1 and type 2 diabetes, dyslipidemia characterized by insulin resistance, erectile dysfunction, cardiovascular disease, and It may be any one selected from the group consisting of ischemic diseases, the liver disease may be any one selected from the group consisting of fatty liver, hepatitis and cirrhosis due to lipid metabolic abnormalities between alcoholic and non-alcoholic.

In addition, the pharmaceutical composition according to the present invention may further comprise an LXR agonist. Preferably, the LXR agonist is an LXRα agonist, and the LXRα agonist is 22 (R) -hydroxycholesterol, 24 (S), 25-epoxycholesterol, 24 (S) -hydroxycholesterol, 27-hydroxycholesterol , Cholesteric acid, T-314407, T-0901317, GW3965, SB742881, GSK 2186, WAY-252623 (LXR-623), WAY-254011, WYE-672, DMHCA (N, N-Dimethyl-3b-hydroxycholenamide) , 4- (3-biaryl) quinoline sulfone, F ₃ MethylAA, acetyl-podocarpic dimer (APD), podocarpic imide dimer, CRX000541, CRX000864, CRX000929, CRX000823, CRX000987, CRX001093, CRX001094, CRX156651, CRX000909, CRX000908, CRX000369, CRX001045, CRX001046, LN7181, LN7179, LN7172, LN6672, LN7031, LN7033, LN6500, LN6662, Ricardin C (Riccardin C), XL-652, DY136, DY142, DY142, DY142 6a-dihydroxy-5b-cholanoic acid-N-methyl-N-methoxy-24-amide), but is not limited thereto and used as an LXRα agonist. But it may be as long as they would any.

The pharmaceutical composition according to the present invention preferably contains 0.1 to 50.0 parts by weight of TRAP80 modifier based on 100 parts by weight of the pharmaceutical composition. If the TRAP80 regulator is included outside the content range, it is not possible to selectively inhibit the transcription of SREBP-1c, which is an important regulator of de novo fatty acid synthesis, and thus cannot obtain a desired effect in the present invention.

In addition, the pharmaceutical composition of the present invention may further comprise a suitable carrier, excipient or diluent commonly used in the manufacture of the pharmaceutical composition.

Carriers, excipients or diluents that may be included in the pharmaceutical composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium Silicates, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The pharmaceutical composition according to the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions according to a conventional method .

In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid form preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, which form at least one excipient such as starch, calcium carbonate, sucrose or Prepare by mixing lactose, gelatin and the like.

In addition to simple excipients, lubricants such as magnesium styrate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included .

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of suppository bases include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like.

The dosage of the TRAP80 modulator used in the present invention can be increased or decreased depending on the route of administration, the severity of the disease, sex, weight, age and the like.

The pharmaceutical composition may be administered to mammals such as rats, mice, livestock, humans, and the like in a variety of routes. All modes of administration can be expected, for example by oral, rectal or intravenous, nasal, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

In addition, the present invention comprises the steps of contacting the candidate drug with blood or tissues of patients with metabolic syndrome; And evaluating whether the interaction between the LXR and TRAP80 is inhibited by the candidate drug.

In addition, the present invention comprises the steps of obtaining a TRAP80 expression profile from a sample of a patient; And evaluating the TRAP80 expression profile by comparing with the TRAP80 expression profile obtained from a sample of a normal person.

In addition, the present invention provides a method for reducing the side effects of LXR agonists, characterized in that by administering a TRAP80 modulator inhibits the interaction between LXR and TRAP80.

The TRAP80 modulator may be any one selected from the group consisting of siRNA that inhibits TRAP80 expression and an antibody that specifically binds to TRAP80, and the TRAP80 modulator may selectively inhibit transcription of SREBP-1c.

The LXR agonist is an LXRα agonist, for example 22 (R) -hydroxycholesterol, 24 (S), 25-epoxycholesterol, 24 (S) -hydroxycholesterol, 27-hydroxycholesterol, cholesterol Iksan, T-314407, T-0901317, GW3965, SB742881, GSK 2186, WAY-252623 (LXR-623), WAY-254011, WYE-672, DMHCA (N, N-Dimethyl-3b-hydroxycholenamide), 4- ( 3-biaryl) quinoline sulfone, F ₃ MethylAA, acetyl-podocarpic dimer (APD), podocarpic imide dimer, CRX000541, CRX000864, CRX000929, CRX000823, CRX000987, CRX001093, CRX001094, CRX156651, CRX000909, CRX000908, CRX000369, CRX001045, CRX001046, LN7181, LN7179, LN7172, LN6672, LN7031, LN7033, LN6500, LN6662, Ricardin C (Riccardin C), XL-652, DY136, DY142, and hypocollamide (3a, dia, 6-hydroxy) 5b-cholanoic acid-N-methyl-N-methoxy-24-amide).

Side effects of the LXR agonist mean unwanted effects on fatty acid metabolism, including an increase in serum triglycerides and hepatic triglycerides.

The pharmaceutical composition for treating or preventing metabolic syndrome comprising the TRAP80 modulator as an active ingredient effectively inhibits the transcription of SREBP-1c by effectively containing the TRAP80 modulator as an active ingredient to effectively treat metabolic syndrome such as arteriosclerosis, fatty liver, etc. Can be prevented. It can also inhibit fatty acid metabolism activities such as serum triglycerides and hepatic triglycerides, which are unwanted effects of LXR agonists, thus enhancing the desirable effects of LXR agonists such as increasing serum HDL and reducing arteriosclerosis. Therefore, the effect as a therapeutic agent of metabolic syndrome, such as arteriosclerosis or fatty liver, by an LXR agonist can be improved further.

1 is a protein that can interact with LXRa LBD from HepG2 cells, A shows a schematic diagram of LXRa LBD fused to GST, B shows overexpression and purification of GST-LXRa LBD, C is Ligand-dependent interactions between several nuclear proteins obtained from LXRa LBD and HepG2 cells are shown, and D shows Western blot analysis of GST-LXRa LBD interactor.
Figure 2 analyzes the interaction between LXRa and the nuclear protein binding to it, A shows the expression of Flag-LXRa in HEK293T cells transfected with Flag-LXRa, B is anti- from cells transfected with Flag-LXRa The result of immunoprecipitation and Western blot analysis between LXRa and interacting protein using the flag antibody, C is the result of GST pull-down analysis between GST-LXRa LBD and ³⁵ [S] -methionine labeled interacting agent.
3 relates to the regulation of LXRE-containing promoter activity by TRAP220 and TRAP80.
Figure 4 shows the chromatin immunoprecipitation analysis of the SREBP-1c and ABCA1 promoter.
5 analyzes the different effects of TRAP80 siRNA on the levels of SREBP-1c and ABCA1 transcripts by universal RT-PCR and quantitative real-time PCR.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the present invention is not limited by these examples.

Example 1 Identification of Proteins Interacting with LXRa LBD

For the identification of proteins interacting with LXRa LBD, GST pull-down analysis was performed according to previously known methods (Genes Dev. 1998; 12: 1787-1800). That is, before reacting with the HepG2 nuclear extract, glutathione-sepharose beads-fixed GST and GST-LXRα LBD were adjusted to equivalent molarity by quantification using SDS-PAGE. Bound with binding solution [20 mM HEPES (pH 7.6), 20% glycerol, 0.2 mM EDTA, 180 mM KCl, 1 mM DTT, 0.05% Nonidet P-40, 0.5 mM PMSF, 10 μM T0901317] and washed solution [20 mM HEPES (pH 7.6) ), 20% glycerol, 0.2 mM EDTA, 180 mM KCl, 1 mM DTT, 0.1% Nonidet P-40, 0.5 mM PMSF], and then elute the sample with 0.2% Sarkosyl dissolved in the wash buffer, and 4-20%. Separation by SDS-PAGE (BioRad) and staining using Silver Stain Plus (Biorad) or Western blot were used.

First, GST-fusion LXRa LBD (amino acid residues 159-445) was expressed and purified using glutathione-Sepharose beads as shown in FIGS. 1A and 1B. After reacting the HepG2 nuclear extract in the presence or absence of T0901317, a synthetic LXR ligand, the interacting protein was eluted from the beads and analyzed through SDS-PAGE and silver staining, specific to GST-LXRa LBD as shown in FIG. 1C. Five proteins bound to each other were identified.

Example 2 Analysis of LXRa LBD Binding Protein Using a Mass Spectrometer

LXRa LBD binding protein was applied to gel trypsin digestion in silver sliced gel slices. Extracted peptides are desalted with Zip-tip ^TM (Millipore), resuspended in matrix and matrix-assisted laser desorption using Voyager Biospectrometry (Applied Biosystems, Foster City, Calif.) Ionization-time of flight (MALDI-TOF) analysis was performed to obtain peptide mass fingerprints. The obtained data was processed using Mascot program (www.matrixscience.com).

The five proteins found in Example 1 were identified using MALDI-TOF analysis, and further analysis of the two proteins (TRAP220 and TRAP80) that interacted directly with LXR was shown in Table 1. Presented. In addition, the protein was re-validated by Western blot analysis results as shown in FIG. 1D.

protein Score ^* Prediction ^† Number of peaks identified ^‡ Matched mass ^§ Protein mass Accession No. TRAP220 / DRIP205 34 6.3 9 23.7% 168 Q15648 TRAP80 / CRSP77 81 1.2e ^-4 16 19.3% 73 Q9NVC6

^* Protein score indicates the degree of statistical significance of the match, scores above 65 are significant values, and ^† Predictive value is the likelihood that the analyte will only be identified as a corresponding protein above the corresponding protein score by chance, which is a Blast search. results and E- equivalent value in, the more unexpected the lower score than the protein and significantly, ^‡ and can be identified peaks of the mass values match, ^§ matched masses that match the mass of the number of irradiated mass values The percentage of the number of values.

Example 3 Immunoprecipitation and Western Blot Analysis

HEK293T cells transfected with Flag-LXRa were treated for 2 hours in 1 mM T0901317, washed in phosphate buffer, 10 in Lysis buffer consisting of 1% NP-40, 10% glycerol and 50 mM Tris-Cl, pH 7.4. Dissolve on ice for minutes. Immunoprecipitation assay by adding 6 μg anti-Flag antibody (F7425, Sigma) and reacting overnight at 4 ° C. in IP buffer (180 mM KCl, 1% NP-40, 10% glycerol and 50 mM Tris-Cl, pH 7.4) Was performed. Immune complexes were precipitated with protein G-agarose (sc-2002, Santa Cruz), washed with IP buffer and resuspended in reduced sample buffer. Proteins were separated by SDS-PAGE, transferred to PVDF membrane (QBIOGENE) and detected using antibodies against TRAP220 (sc-5334), TRAP80 (ARP34192_P050, Aviva Systems Biology) and Flag.

As a result, all five proteins were identified in vivo , and the results for TRAP220 and TRAP80 are shown in Figure 2B.

In addition, GST pull-down analysis between the GST-LXRa LBD and the [ ³⁵ S] -methionine labeled interactor revealed that only two proteins, TRAP220 and TRAP80, directly interacted with LXRa LBD as shown in FIG. 2C. In particular, [ ³⁵ S] -TRAP220 bound to GST-LXRa LBD interacted ligand dependently, while [ ³⁵ S] -TRAP80 interacted separately with ligand.

Example 4 Conformal Infection and Luciferase Reporter Analysis

To examine the functional relevance of TRAP220 and TRAP80 in the LXRa assay, cotransfection and luciferase reporter assays were performed. Specifically, while transfecting the Luciferase reporter plasmid containing the synthetic LXRE promoter into HepG2 cells, expression plasmids such as LXRa, RXRa, TRAP220, TRAP80 and the like were cotransfected. After 24 hours, dimethyl sulfoxide (DMSO) or T0901317 was treated, and further incubated for 24 hours, cells were lysed and luciferase activity was measured using a luminometer (Berthold Technologies). As a result, TRAP220 and TRAP80 induced ligand-dependent transcriptional activity of the synthetic LXRE promoter as shown in FIG. 3A. This coactivator function was also identified on two major natural promoters, including the LXRE portion of ABCA1 or SREBP-1c, as shown in FIGS. 3B and 3C.

TRAP220 and TRAP80 did not induce transcription in the SREBP-1c LXRE mutant promoter, meaning that the function of the coactivator is specifically dependent on LXR recruitment to LXRE (FIG. 3D). Likewise, neither the ABCA1 LXRE variant promoter was activated by TRAP220 or TRAP80. From these results, it can be seen that the binding of LXR to LXRE of ABCA1 and SREBP-1c promoters is essential for increasing the transcriptional activity of both genes by TRAP220 or TRAP80.

Example 5 Chromatin Immunoprecipitation Assay

To investigate the mechanism of TRAP220 and TRAP80 recruitment to endogenous chromatin LXRE, chromatin immunoprecipitation and PCR analysis was performed on the SREBP-1c and ABCA1 promoter regions.

This example performed a chromatin immunoprecipitation assay with a slight modification of the previously known report ( Methods , 1997; 11: 205-214). That is, 1% formaldehyde cells were treated for 20 minutes at room temperature, followed by 20 minutes at 4 ° C. Cells were then washed with cold phosphate buffer (PBS) and resuspended in Lysis buffer containing 50 mM Tris-HCl (pH 8.0), 1% SDS and 10 mM EDTA. The chromatin dissolved from the cells was prepared by sonication and immunized with antibodies against TRAP220 (sc-5334, Santa Cruz), TRAP80 (ARP34192_P050, Aviva Systems Biology), and acetylated H4 (06-598, Upstate). Settled. The final DNA extract was amplified using primers specific for the SREBP-1c and ABCA1 promoters to obtain a 250 bp product.

As a result, both TRAP220 and TRAP80 were recruited to LXRE of the endogenous SREBP-1c promoter in response to T0901317 as shown in FIG. 4A. In addition, the histone acetylation effect on the recruitment of TRAP220 and TRAP80 to the SREBP-1c promoter was examined. Namely, treatment with histone acetyltransferase inhibitor garcinol inhibited not only TRAP220 and TRAP80 recruitment but also acetylated H4 levels as expected. In particular, TRAP80 was more sensitive to the acetylation status of the SREBP-1c promoter than TRAP220. The recruitment of TRAP80 to the ABCA1 promoter was less responsive to the acetylation states of the ligand and histone, while TRAP220 showed a similar recruitment pattern to both ABCA1 and SREBP-1c promoters (FIG. 4B). TRAP80 showed different effects on ABCA1 and SREBP-1c in terms of chromatin.

Example 6 Different Regulatory Effects of LXR Target Genes by TRAP80

As previously discussed, in luciferase reporter assay, TRAP80 upregulated the transcriptional activity of LXRa on a ligand dependent basis on ABCA1 and SREBP-1c promoters (FIG. 3). TRAP80, on the other hand, increased recruitment to the SREBP-1c promoter in response to the ligand, but recruitment to the ABCA1 promoter was not associated with the presence of the ligand (FIG. 4). To demonstrate this difference, TRAP80 siRNA (Santa Cruz, sc-38575) was transfected into HepG2 cells, T0901317 was treated for 18 hours and total RNA was isolated using Trizol (Invitrogen). The isolated RNA was treated with DNaseI (10u / mg of total DNA), and cDNA was synthesized using Moloney murine leukemia virus reverse transcriptase (Promega). After using the resulting cDNA as a template, the SREBP-1c and ABCA1 transcript levels were determined by performing quantitative real-time PCR of the SREBP-1c and ABCA1 genes using the SYBR green PCR master mix and ABI PRISM 7000 equipment (FIG. 5). ). In this case, scrambled siRNA was used as a negative control group, and PCR primers were designed using Primer Express 2.0 software.

As a result, as shown in Figure 5 TRAP80 siRNA significantly inhibited the production of SREBP-1c transcripts with or without T0901317. ABCA1 transcript levels, on the other hand, did not show significant changes by TRAP80 siRNA. These results indicate that SREBP-1c and ABCA1, which are target genes of LXR, are regulated differently by TRAP80, and that inhibition of TRAP80 can selectively inhibit SREBP-1c of LXR target genes.

Claims (16)

A pharmaceutical composition for preventing or treating metabolic syndrome, comprising a TRAP80 modulator selected from the group consisting of siRNAs that inhibit TRAP80 expression and antibodies specifically binding to TRAP80. delete The pharmaceutical composition for preventing or treating metabolic syndrome according to claim 1, wherein the TRAP80 modulator selectively inhibits transcription of SREBP-1c. The method according to claim 1, wherein the metabolic syndrome is arteriosclerosis, liver disease, hyperlipidemia, hypercholesterolemia, obesity, hypertension, hyperinsulinemia, hyperglycemia, type 1 and type 2 diabetes, dyslipidemia characterized by insulin resistance, erectile dysfunction, Pharmaceutical composition for preventing or treating metabolic syndrome, characterized in that selected from the group consisting of cardiovascular diseases and ischemic diseases. The pharmaceutical composition for preventing or treating metabolic syndrome according to claim 4, wherein the liver disease is selected from the group consisting of fatty liver, hepatitis and cirrhosis due to abnormal lipid metabolism between alcoholic and non-alcoholic liver. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is 22 (R) -hydroxycholesterol, 24 (S), 25-epoxycholesterol, 24 (S) -hydroxycholesterol, 27-hydroxycholesterol, cholesteroic acid, T- 314407, T-0901317, GW3965, SB742881, GSK 2186, WAY-252623 (LXR-623), WAY-254011, WYE-672, DMHCA (N, N-Dimethyl-3b-hydroxycholenamide), 4- (3-biaryl Quinoline sulfone, F ₃ MethylAA, acetyl-podocarpic dimer (APD), podocarpic imide dimer, CRX000541, CRX000864, CRX000929, CRX000823, CRX000987, CRX001093, CRX001094, CRX156651, CRX000909, CRX000908, CRX000909 CRX001045, CRX001046, LN7181, LN7179, LN7172, LN6672, LN7031, LN7033, LN6500, LN6662, Ricardin C, XL-652, DY136, DY142 and hypocollamide (3a, 6a-dihydroxy-5b-cholanoic acid -N-methyl-N-methoxy-24-amide) pharmaceutical composition for the prevention or treatment of metabolic syndrome, further comprising an LXR agonist selected from the group consisting of water. delete Contacting the candidate drug with blood or tissue of a metabolic syndrome patient; And evaluating whether the interaction between LXR and TRAP80 is inhibited by the candidate drug. Obtaining a TRAP80 expression profile from a patient's sample; And evaluating the TRAP80 expression profile by comparing it with a TRAP80 expression profile obtained from a sample of a normal person. delete delete delete delete delete delete delete

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