KR101625333B1 - Pharmaceutical composition comprising inhibitor of Carbohydrate responsive element-binding protein(ChREBP) expression or activity for prevention or treatment of type 1 diabetes - Google Patents

Abstract

The present invention relates to a pharmaceutical composition comprising an inhibitor of expression or activity of ChREBP as an active component for preventing or treating type 1 diabetes. The pharmaceutical composition according to the present invention can be used in prevention or treatment of type 1 diabetes by protecting beta cells by inhibiting expression of TXNIP involved in death of the pancreatic beta cells by inhibiting expression or activity of ChREBP which is a transfer factor of TXNIP.

Description

The present invention relates to a pharmaceutical composition for preventing or treating type 1 diabetes comprising an expression or activity inhibitor of ChREBP as an active ingredient. }

The present invention relates to a pharmaceutical composition for the prevention or treatment of type 1 diabetes, comprising as an active ingredient an expression or activity inhibitor of ChREBP.

Type 1 diabetes is a disease caused by the destruction of the pancreatic β-cells of the pancreas that secrete insulin. It is also called insulin-dependent diabetes because it requires continuous administration of insulin after onset, Lt; / RTI > The cause is unknown, but it is known to be caused by genetic, environmental, viral, and chemical agents. Symptoms usually appear suddenly and severe symptoms can include increased thirst, frequent urination, weight loss, severe hunger, vomiting, abdominal pain, or fatigue.

Type 1 diabetes accounts for 5-10% of the overall incidence of diabetes and is reported worldwide to occur in 80,000 children every year, but the exact number of patients is unknown. It is estimated that there are between 1 million and 3 million patients in the United States alone. In Korea, the incidence rate is gradually increasing due to one patient per 100,000 patients.

The destruction of pancreatic beta cells in type 1 diabetes results in insulin deficiency and eventually destroys glucose homeostasis in the body. Because insulin lowers blood sugar by transporting glucose in the blood into cells and by converting glucose into glycogen in the liver, the insulin deficiency resulting from the destruction of beta cells leads to glucose homeostasis in the blood, leading to hyperglycemia and metabolic disorders Lt; / RTI > Recently, oxidative stress in pancreatic beta cells has been reported to affect beta cell death and function loss in the progression of type 1 diabetes.

The mTOR (mTOR complex 1) and mTORC2 (mTOR complex 2), which interact with various proteins, play an important role in the first step of nutrient sensing and glucose homeostasis maintenance. ). MTORC1 functions to regulate mRNA transcription, protein synthesis, lipid synthesis, and autophagy, while mTORC2 functions to regulate cell survival, cell cycle progression, and insulin-induced assimilation It is known to be involved in the regulation of insulin-induced anabolism.

Recently, although many studies have been carried out on the proteins of each mTOR complex or sub-signaling in glucose metabolism, the key factors related to the survival and function of beta cells and the direct role of mTOR are not known.

Therefore, the present inventors sought to investigate the role of mTOR in controlling the survival of pancreatic beta cells and thus the insulin secretion, and the novel target (ChREBP) and its mechanism of action, which are key to the role of mTOR.

The inventors of the present invention have studied the direct role of mTOR having the function of regulating insulin secretion and the survival of pancreatic beta cells. As a result, when mTOR is deleted or its expression is inhibited, expression of ChREBP is increased and binding of Mlx and ChREBP And the expression of TXNIP, which is involved in beta cell apoptosis, is increased. That is, the present inventors have completed the present invention by discovering that mTOR competitively inhibits the transcription activity of ChREBP by binding to ChlBP with Mlx, thereby inhibiting the expression of TXNIP involved in beta cell death.

Accordingly, it is an object of the present invention to provide a pharmaceutical composition for the prevention or treatment of type 1 diabetes, which comprises as an active ingredient an expression or activity inhibitor of ChREBP.

However, the technical problem to be solved by the present invention is not limited to the above-mentioned problems, and other matters not mentioned can be clearly understood by those skilled in the art from the following description.

In order to accomplish the above object, the present invention provides a pharmaceutical composition for preventing or treating type 1 diabetes comprising, as an active ingredient, an expression or activity inhibitor of Carbohydrate-responsive element-binding protein (ChREBP) do.

In one embodiment of the present invention, the expression or activity inhibitor of ChREBP is mTOR (mammalian Target Of Rapamycin) gene or mTOR protein.

In another embodiment of the present invention, the mTOR gene is characterized by being composed of the nucleotide sequence of SEQ ID NO: 1.

In another embodiment of the present invention, the mTOR protein is characterized by being composed of the amino acid sequence of SEQ ID NO: 2.

In another embodiment of the present invention, the composition inhibits expression of TXNIP (Thioredoxin-interacting protein) by inhibiting the expression or activity of ChREBP.

The present invention also provides a method of preventing or treating type 1 diabetes, comprising the step of administering the composition to a subject.

The present invention also provides the use of the composition for the prevention or treatment of type 1 diabetes.

The pharmaceutical composition according to the present invention inhibits the expression or activity of ChREBP, which is a transcription factor of TXNIP, to inhibit the expression of TXNIP, which is involved in pancreatic beta cell death, and thus has a beta cell protection effect. Thus, . ≪ / RTI >

FIG. 1A shows the expression of TXNIP mRNA in the inhibition of mTOR expression under streptozotocin (STZ) treatment conditions.
FIG. 1B shows the expression of TXNIP protein in the presence of inhibition of mTOR expression under streptozotocin (STZ) treatment conditions.
FIG. 2A shows the expression of TXNIP mRNA in pancreatic beta cells of mice lacking mTOR under glucose stimulation conditions.
FIG. 2B shows the expression of TXNIP mRNA in the presence of inhibition of mTOR expression under glucose stimulation conditions.
Figure 2c shows the expression of TXNIP protein in the presence of mTOR inhibition under glucose stimulation conditions.
FIG. 3A shows the expression of ChREBP mRNA expression in the inhibition of mTOR expression under glucose stimulation conditions.
FIG. 3B shows the expression of ChREBP protein in the inhibition of mTOR expression under glucose stimulation conditions.
FIG. 3C shows the increase in the binding affinity of the ChREBP and TXNIP promoter ChoRE sites in the inhibition of mTOR expression under glucose stimulation conditions.
FIG. 3D shows the increase in binding affinity between the ChREBP and TXNIP promoter ChoRE sites under streptozotocin (STZ) treatment conditions.
Figure 4a shows the increase in binding of Mlx and ChREBP, the transcriptional regulators of TXNIP under streptozotocin (STZ) treatment conditions.
Figure 4b shows the increased binding of Mlx and ChREBP, the transcriptional regulators of TXNIP under glucose stimulation conditions.
Fig. 4C shows the remarkable increase in the binding of Mlx and ChREBP upon inhibition of mTOR expression.
Figure 5 depicts the modulation of TXNIP expression by mTOR.
FIG. 6A shows fluorescence images and quantitative results showing the increase of beta cell death upon mTOR deletion under streptozotocin (STZ) treatment conditions by the TUNEL assay.
FIG. 6B shows the expression of apoptosis-inducing-related protein upon inhibition of mTOR expression under streptozotocin (STZ) treatment conditions.

The present invention provides a pharmaceutical composition for the prevention or treatment of type 1 diabetes comprising as an active ingredient an expression or activity inhibitor of ChREBP.

The type 1 diabetes of the present invention refers to a form of diabetes caused by the destruction of the Langerhans beta cells of the pancreas producing insulin and the failure of normal glucose storage by insulin.

As used herein, the term " prophylactic " means any action that inhibits type 1 diabetes or delays its onset by administration of a pharmaceutical composition according to the present invention.

As used herein, the term " treatment " means any action that improves or alters the symptoms of type 1 diabetes by the administration of the pharmaceutical composition of the present invention.

The term " activity inhibitor " used in the present invention means that the function of the target protein is degraded, and thus the function of the target protein is undetectable or present at a meaningless level.

The expression or activity inhibitor of the ChREBP of the present invention may be an mTOR gene or an mTOR protein.

The mTOR gene may be composed of the nucleotide sequence of SEQ ID NO: 1 or 3 derived from human or mouse, and the mTOR protein may be composed of the amino acid sequence of SEQ ID NO: 2 or 4 derived from human or mouse. And more preferably the human-derived base sequence of SEQ ID NO: 1 or the amino acid sequence of SEQ ID NO: 2, but is not limited thereto.

The ChREBP, also referred to as MLX-interacting protein-like (MLXIPL), interacts with Mlx and is a protein that functions as a transcription factor for Myc, Max, and Mad family proteins. The ChREBP protein functions to activate transcription by binding to a carbohydrate response element (ChoRE) present in the promoter of triglyceride synthesis genes in a glucose-dependent manner.

Mlx (Max like protein) is one of the basic helix-loop-helix leucine zipper (bHLH-Zip) transcription factor family proteins and is involved in cell proliferation, crystallization and differentiation by forming a heterodimer with Mad protein. The Mlx protein is known to interact with MNT, MXD1, and ChREBP (MLXIPL).

Hereinafter, the role of mTOR as an inhibitor of ChREBP expression or activity was identified through the examples of the present invention.

In one embodiment of the present invention, in order to examine the role of mTOR in the survival of beta cells and the regulation of insulin secretion, the expression level of TXNIP was examined under mTOR deletion or expression inhibition under streptozotocin treatment or glucose stimulation, And increased protein expression (see Example 2).

The TXNIP (Thioredoxin-interacting protein) is known to be a major regulator of the redox system that induces the production of active oxygen species and apoptosis in beta cells. Overexpression of TXNIP in pancreatic beta cells promotes apoptosis and induces resistance to streptozotocin-mediated type 1 diabetes through the induction of Akt / Bcl-xL signaling when deficient.

In another embodiment of the present invention, the expression level of ChREBP, which is known as a transcription factor of TXNIP, is increased when mTOR deletion or expression is inhibited under streptozotocin treatment or glucose stimulation conditions. As a result, expression of mRNA and protein of ChREBP is increased Respectively. In addition, it was confirmed that the decrease in mTOR expression further increased the binding affinity between ChREBP and the TXRE promoter ChoRE site (see Example 3).

In another embodiment of the present invention, mTOR binds to ChREBP competitively with Mlx by confirming that the binding of Mlx to ChREBP, which acts as a transcriptional regulator of TXNIP, is increased in the beta cells in which mTOR expression is inhibited It was found that the transcription of TXNIP was suppressed (see Example 4).

In another embodiment of the present invention, the beta-cell protection effect of mTOR was directly examined based on the results of the above examples. As a result, when mTOR was deleted or its expression was inhibited under streptozotocin treatment conditions, (See Example 5). ≪ tb > < TABLE >

The pharmaceutical composition according to the present invention contains as an active ingredient an expression or activity inhibitor of ChREBP and may include a pharmaceutically acceptable carrier. Such pharmaceutically acceptable carriers are those conventionally used in the field of application and include, but are not limited to, saline, sterile water, Ringer's solution, buffered saline, cyclodextrin, dextrose solution, maltodextrin solution, glycerol, ethanol, And may further contain other conventional additives such as antioxidants and buffers as needed. In addition, it can be formulated into injectable formulations, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like by additionally adding diluents, dispersants, surfactants, binders, lubricants and the like. Suitable pharmaceutically acceptable carriers and formulations can be suitably formulated according to the respective ingredients using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA. The pharmaceutical composition of the present invention is not particularly limited to a formulation, but may be formulated into injections, inhalants, external skin preparations, and the like.

The pharmaceutical composition of the present invention may be administered orally or parenterally (for example, intravenous, subcutaneous, skin, nasal, or airway) according to the desired method, The type of administration, the route of administration, and the time, but may be suitably selected by those skilled in the art.

The composition according to the invention is administered in a pharmaceutically effective amount. In the present invention, " pharmaceutically effective amount " means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and an effective dosage level is determined depending on the type of disease, severity, , Sensitivity to the drug, time of administration, route of administration and rate of release, duration of treatment, factors including co-administered drugs, and other factors well known in the medical arts. The composition according to the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered singly or in multiple doses. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without side effects, which can be easily determined by those skilled in the art.

Specifically, the effective amount of the composition according to the present invention may vary depending on the age, sex, and body weight of the patient. In general, 0.001 to 150 mg, preferably 0.01 to 100 mg, It may be administered one to three times a day. However, the dosage may be varied depending on the route of administration, the severity of obesity, sex, weight, age, etc. Therefore, the dosage is not limited to the scope of the present invention by any means.

The composition of the present invention can be used variously for medicines, foods and beverages for prevention and improvement of type 1 diabetes mellitus, and can be used in powder, granule, tablet, capsule or beverage form.

In another aspect of the present invention, the present invention provides a method of preventing or treating type 1 diabetes comprising administering the composition to a subject.

As used herein, the term " individual " refers to a subject in need of treatment for a disease, and more specifically refers to a human or non-human primate, mouse, rat, dog, cat, horse, Of mammals.

In another aspect of the present invention, the present invention provides the use of the composition for the prevention or treatment of type 1 diabetes.

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the following examples.

[Example]

Example 1. Experimental preparation and method

1-1. mTOR knock-out mouse preparation

In order to construct a mTOR knock-out mouse specifically for pancreatic beta cells, a ^transgenic mouse having mTOR ^{flox / flox} mouse and a rat (rat insulin promoter) cre recombinase was crossed Beta cell specific mTOR knock-out mice were prepared and the mice were purchased from Jackson Laboratory (Bar Harbor, Maine, USA). As a control for beta cell-specific mTOR knock-out mice, mTOR ^{flox / flox} mice with normal mTOR gene were used.

1-2. Cell culture and siRNA transfection

INS-1, a rat-derived insulinoma beta cell, RPMI 1640 medium containing 10% FBS (fetal bovine serum), 1 mM sodium pyruvate, 2 mM L-glutamine, 10 mM HEPES, and 0.05 mM 2-mercaptoethanol supplied from Christopher Newgard (Duke University, Durham, (Welgene, Daegu, South Korea) under 5% CO ₂ and 37 ° C.

In order to knock down the expression of mTOR at the cellular level, the INS-1 cells were plated on a 6-well plate and after 24 hours, mTOR siRNA (Qiagen, Hilden, Germany) or non-silencing siRNA (Qiagen) 10 nM for 72 hours using DharmaFECT1 reagent (Dharmacon, Lafayette, CO, USA). Streptozotocin (STZ) was treated for 24 hours after transfection and the cells were recovered and tested. The target sequences of the mTOR siRNA used are shown in Table 1 below.

order SEQ ID NO: mTOR siRNA 5'-CAGGCCTATGGTCGAGATTTA-3 ' 5

1-3. Isolation of mouse islets

The pancreas was isolated from 8 week old mTOR ^{flox / flox} mice and beta cell specific mTOR knock-out mice, and the pancreas was separated from HBSS (0.8 ^g / ml collagenase P (Roche Diagnostics, Indianapolis, IN, USA) Welgene) solution and incubated at 37 ° C for 15 minutes with regular shaking. The solution in which the pancreatic tissue was degraded was sieved through a mesh and density gradient centrifugation was performed using Ficoll gradients (Biochrom AG, Berlin, Germany) to isolate the intermediate layer and isolate the islets under a microscope. The isolated islets were then cultured in RPMI 1640 medium containing 10% FBS and penicillin / streptomycin overnight at 37 ° C in 5% CO ₂ .

1-4. Glucose stimulation

The INS-1 cells were transfected with mTOR siRNA or non-silencing siRNA by the method of Example 1-2 for 72 hours, and the Krebs-Ringer buffer (KRBH buffer: 135 mM NaCl, 3.6 mM KCl , 0.5 mM NaH ₂ PO ₄ , 0.5 mM MgSO ₄ , 1.5 mM CaCl ₂ , 5 mM NaHCO ₃ , 10 mM HEPES, and 0.1% BSA, pH 7.4) at 37 ° C for 1 hour. Cells were then incubated with KRBH buffer containing 3 mM (low concentration) or 25 mM (high concentration) of glucose for 1 hour at 37 ° C and then the cells were recovered.

Ten islets of similar sizes separated from the mice were pre-incubated with Krebs-Ringer buffer containing 3 mM glucose in the same manner as the above INS-1 cells. After incubation with Krebs-Ringer buffer containing 3 mM (low concentration) or 16.7 mM (high concentration) of glucose, the experiment was carried out.

1-5. Real-time polymerase chain reaction

Total RNA was isolated from Tissol (Invitrogen, Carlsbad, CA, USA) from INS-1 cells transfected with isolated beta-cells or mTOR siRNA and non-silencing siRNA from mouse. CDNA was synthesized by performing Reverse Transcription Polymerase Chain Reaction (RT-PCR) using SuperScript II reverse transcriptase (Invitrogen) and random hexamer primers. Real-time quantitative PCR (qRT-PCR) was performed using SYBR Green PCR Master Mix (Applied Biosystems) with a primer specific for the gene to be expressed in the DNA precipitated from the cDNA or ChIP analysis. Respectively. The expression level of mRNA per sample was corrected with GAPDH mRNA and the gene specific primer sequences used are shown in Table 2 below.

gene direction order SEQ ID NO: rat ChREBP-α forward 5'-CGA CAC TCA CCC GCC TCT TC-3 ' 6 reverse 5'-TTG TTC AGC CGA ATC TTG TC-3 ' 7 rat ChREBP-β forward 5'-TCT GCA GAT CGC GCG GAG-3 ' 8 reverse 5'-CTT GTC CCG GCA TAG CAA C-3 ' 9 rat TXNIP forward 5'CAA GTT CGG CTT TGA GCT TC-3 ' 10 reverse 5'ACG ATC GAG AAA AGC CTT CA-3 ' 11 mouse TXNIP forward 5 'TAT GTA CGC CCC TGA GTT CC-3' 12 reverse 5'GCT CAC TGC ACG TTG TTG TT-3 ' 13

1-6. Immunoblotting and immunoprecipitation

To the INS-1 cells transfected with beta cells or mTOR siRNA and non-silencing siRNA isolated by the method of Example 1-3, RIPA buffer (150 mM NaCl, 50 mM Tris-Cl, pH 7.4 , 1 mM EDTA (ethylenediaminetetraacetic acid), 1% NP-40 (NonidetP-40), 0.25% Na-deoxycholate, 0.1 M NaF and 2 mM Na ₃ VO ₄ ) and protease inhibitors (Roche Diagnostics) . SDS-polyacrylamide gel electrophoresis, transfer, and blocking were performed according to methods known in the art. The primary antibodies used to confirm the expression levels of the respective proteins are as follows.

Antibodies to pT6K and pS6 (S235 / 236), p-S6 (S240 / 244), p-Akt (S473) and pS6K , Mlx, and β-actin antibodies were purchased from Santa-Cruz Biotechnology (USA).

Immunoprecipitation (IP) analysis was performed according to the following method. The protein lysate was reacted with 2 μg of the primary antibody overnight at 4 ° C and the protein / antibody complex was reacted with protein A or G-Sepharose (Amersham, Piscataway, NJ, USA) for 2 hours in the cold room. After the reaction, the cells were washed three times with IP buffer, and the immunoprecipitates were obtained by centrifugation, followed by denaturation by boiling for 5 minutes. Subsequently, the precipitated protein was identified by SDS-polyacrylamide gel electrophoresis and immunoblotting.

1-7. ChIP analysis

Cells transfected with mTOR siRNA or non-silencing siRNA by the method of Example 1-2 and cells treated with glucose by the method of Example 1-4 were treated with 1% formaldehyde and fixed for 10 minutes And glycine was added to a final concentration of 0.125 M to complete the immobilization reaction. The immobilized cells were washed with phosphate buffered saline (PBS), lysed with SDS lysis buffer, sonicated for 30 seconds with a Fisher Scientific Sonic Dismembrator 500, and the intracellular DNA was cut into 200-300 base pairs. Then, the DNA / protein complex was immobilized by immersing the protein / DNA complex in 60 μl of 50% (vol./vol.) Protein A or G agarose Amersham) / salmon sperm DNA slurry for 1 hour at 4 ° C and washed. The cross-linked DNA and protein were annealed in Tris-HCl pH 6.5, 5 M NaCl, and 0.5 M EDTA for 4 hours at 65 ° C, and the DNA was extracted with phenol / CHCl ₃ and precipitated with ethanol. Real-time polymerase chain reaction was performed using the primers shown in Table 3 below and quantified.

gene direction order SEQ ID NO: rat TXNIP promoter forward 5'-CGC ACC CGA ACA ACA ACC AT-3 ' 14 reverse 5'-GGC AAA GGC CGA GGG CGAA-3 ' 15

Example 2. Confirmation of increase of TXNIP expression by mTOR deletion in beta cells

2-1. Confirmation of increased TXNIP expression by mTOR deletion in type 1 diabetic condition

In order to examine the molecular mechanism by which mTOR regulates the survival of insulin secretion and the survival of beta cells, INS-1 cells, which were inhibited by mTOR expression by the method of Example 1-2, were treated with streptozotocin, And TXNIP mRNA and protein expression levels, which are major regulators of the redox system, induce apoptosis and cell death.

As a result, as shown in Figs. 1A and 1B, mRNA and protein expression of TXNIP was increased in INS-1 cells (si-mTOR) in which the expression of mTOR was inhibited in the group treated with STZ compared to the control group (NS) Respectively. These results suggest that mTOR inhibits the expression of TXNIP under STZ-induced type 1 diabetes mellitus.

2-2. Increased expression of TXNIP by mTOR deletion in glucose stimulation conditions

Based on the results of Example 2-1 above, mRNA and protein expression levels of TXNIP were verified upon mTOR deletion or expression inhibition of beta cells under glucose stimulation conditions.

As a result, as shown in FIG. 2A, the islet beta cells isolated from the beta cell-specific mTOR knock-out mouse as compared to the mTOR ^{flox / flox} mouse (mTOR + / +) in both cases of low glucose and high concentration treatment (mTOR - / -) increased the expression of TXNIP protein. In addition, as shown in FIGS. 2B and 2C, the expression of TXNIP mRNA and protein was also observed in INS-1 cells in which mTOR expression was inhibited, irrespective of glucose concentration. These results suggest that mTOR inhibits the expression of TXNIP under glucose stimulation conditions.

Overall, mTOR regulates oxidative stress by regulating TXNIP expression in beta cells.

Example 3. Confirmation of increase of TXNIP transcription by deletion of mTOR

Glucose acts as a major inducer of TXNIP expression in islets of INS-1 cells and human pancreas, and ChREBP is a transcription factor that regulates the expression of TXNIP in a variety of cells. Based on these results, we confirmed that mTOR regulates the expression of TXNIP by controlling the transcription of ChREBP-mediated TXNIP in beta cells. For this, mRNA and protein expression levels of ChREBP in INS-1 cells inhibited the expression of mTOR under glucose stimulation conditions were observed.

As a result, as shown in FIGS. 3A and 3B, in the case of cells (si-mTOR) in which mTOR expression was inhibited compared to control cells (NS) in which mTOR was normally expressed regardless of glucose stimulation concentration, mRNA and protein expression of ChREBP And the expression of TXNIP protein was also increased. In addition, the expression of TXNIP was not increased when the expression of mTOR was inhibited together with the expression of ChREBP, and the expression of TXNIP was decreased even when the expression of ChREBP was inhibited without inhibiting the expression of mTOR. From the above results, it can be seen that mTOR inhibits the expression of TXNIP through ChREBP.

 Next, ChIP (Chromatin immuno-precipitation) assay was performed by the method of Example 1-7 to examine whether mTOR affects the direct TXNIP transcription control of ChREBP, and the binding affinity of ChREBP and TXNIP promoter ChoRE site was observed Respectively.

As a result, as shown in FIG. 3C, the binding affinity of the ChREBP and TXNIP promoter ChoRE sites was significantly increased in cells in which mTOR expression was inhibited (si-mTOR) compared to the control (NS) under glucose stimulation conditions . These results were the same under STZ-treated type 1 diabetic condition as shown in Figure 3d.

Overall, mTOR inhibits the binding of ChREBP to the promoter of TXNIP and thus protects beta cells by inhibiting the formation of inflammasome complexes.

Example 4. Confirmation of inhibition of binding of Mlx and ChREBP by mTOR

Mlx (Max like protein) is known to act as a transcriptional regulator of TXNIP by complexing with ChREBP during glucose stimulation in INS-1 cells. Immunoprecipitation (IP) was performed by the method of Example 1-6 to verify whether mTOR competitively inhibits the binding of Mlx and ChREBP in the control of TXNIP expression.

As a result, as shown in Figs. 4A and 4B, it was confirmed that the binding of ChREBP and Mlx was increased under STZ treatment and glucose stimulation conditions. In addition, through FIG. 4C, it was confirmed that the binding affinity between ChREBP and Mlx was significantly increased in the cell (si mTOR) in which the expression of mTOR was inhibited compared to the control (si NS). Thus, mTOR competitively inhibits the transcriptional activity of ChREBP by binding to ChlBP under STZ treatment and glucose stimulation conditions, and inhibits the expression of TXNIP. The results are shown in FIG.

Overall, deletion of mTOR induced dimerization of ChREBP and Mlx, and the dimer migrated from the cytoplasm to the nucleus and bound to the ChoRE site of the TXNIP promoter, promoting transcription of TXNIP.

Example 5. Confirmation of increase of beta cell death by deletion of mTOR

The results of Examples 2 to 4 above confirm that mTOR inhibits the transcriptional activity of ChREBP under conditions of type 1 diabetes and glucose stimulation, thereby inhibiting the expression of TXNIP involved in the death of beta cells. Therefore, We directly examined whether mTOR is involved in the inhibition of beta cell death in type 1 diabetes mellitus.

For this purpose, pancreas was isolated from each mouse and immunochemical staining was performed. The tissue sections were stained with TUNEL reagent which can confirm the cell death, and as a result, as shown in FIG. 6A, when STZ was administered The number of cells stained with TUNEL reagent was significantly increased in the beta cell-specific mTOR knock-out mouse (- / - STZ) compared to mTOR ^{flox / flox} mouse (+ / + STZ) .

In addition, INS-1 cells transfected with non-silencing siRNA (NS) or mTOR siRNA (si-mTOR) were treated with STZ at 0.5 or 1 mM for 6 hours and then subjected to Western blotting, Expression of apoptosis related proteins was confirmed.

As a result, as shown in FIG. 6C, when the expression of mTOR was inhibited, the expression of cleaved PARP and BAX protein increased in proportion to the concentration of STZ, and the expression of Bcl-2 protein involved in apoptosis inhibition Respectively.

Through these results, we directly confirmed that mTOR is involved in the suppression of beta cell death.

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 invention. There will be. It is therefore to be understood that the above-described embodiments are illustrative in all aspects and not restrictive.

<110> Research & Business Foundation SUNGKYUNKWAN UNIVERSITY <120> Pharmaceutical composition comprising inhibitor of Carbohydrate          responsive element-binding protein (ChREBP) expression or activity          for prevention or treatment of type 1 diabetes <130> MP15-083 <160> 15 <170> KoPatentin 3.0 <210> 1 <211> 7650 <212> DNA <213> Homo sapiens_mTOR <400> 1 atgcttggaa ccggacctgc cgccgccacc accgctgcca ccacatctag caatgtgagc 60 gtcctgcagc agtttgccag tggcctaaag agccggaatg aggaaaccag ggccaaagcc 120 gccaaggagc tccagcacta tgtcaccatg gaactccgag agatgagtca agaggagtct 180 actcgcttct atgaccaact gaaccatcac atttttgaat tggtttccag ctcagatgcc 240 aatgagagga aaggtggcat cttggccata gctagcctca taggagtgga aggtgggaat 300 gccacccgaa ttggcagatt tgccaactat cttcggaacc tcctcccctc caatgaccca 360 gttgtcatgg aaatggcatc caaggccatt ggccgtcttg ccatggcagg ggacactttt 420 accgctgagt acgtggaatt tgaggtgaag cgagccctgg aatggctggg tgctgaccgc 480 aatgagggcc ggagacatgc agctgtcctg gttctccgtg agctggccat cagcgtccct 540 accttcttct tccagcaagt gcaacccttc tttgacaaca tttttgtggc cgtgtgggac 600 cccaaacagg ccatccgtga gggagctgta gccgcccttc gtgcctgtct gattctcaca 660 acccagcgtg agccgaagga gatgcagaag cctcagtggt acaggcacac atttgaagaa 720 gcagagaagg gatttgatga gaccttggcc aaagagaagg gcatgaatcg ggatgatcgg 780 atccatggag ccttgttgat ccttaacgag ctggtccgaa tcagcagcat ggagggagag 840 cgtctgagag aagaaatgga agaaatcaca cagcagcagc tggtacacga caagtactgc 900 aaagatctca tgggcttcgg aacaaaacct cgtcacatta cccccttcac cagtttccag 960 gctgtacagc cccagcagtc aaatgccttg gtggggctgc tggggtacag ctctcaccaa 1020 ggcctcatgg gatttgggac ctcccccagt ccagctaagt ccaccctggt ggagagccgg 1080 tgttgcagag acttgatgga ggagaaattt gatcaggtgt gccagtgggt gctgaaatgc 1140 aggaatagca agaactcgct gatccaaatg acaatcctta atttgttgcc ccgcttggct 1200 gcattccgac cttctgcctt cacagatacc cagtatctcc aagataccat gaaccatgtc 1260 ctaagctgtg tcaagaagga gaaggaacgt acagcggcct tccaagccct ggggctactt 1320 tctgtggctg tgaggtctga gtttaaggtc tatttgcctc gcgtgctgga catcatccga 1380 gcggccctgc ccccaaagga cttcgcccat aagaggcaga aggcaatgca ggtggatgcc 1440 acagtcttca cttgcatcag catgctggct cgagcaatgg ggccaggcat ccagcaggat 1500 atcaaggagc tgctggagcc catgctggca gtgggactaa gccctgccct cactgcagtg 1560 ctctacgacc tgagccgtca gattccacag ctaaagaagg acattcaaga tgggctactg 1620 aaaatgctgt ccctggtcct tatgcacaaa ccccttcgcc acccaggcat gcccaagggc 1680 ctggcccatc agctggcctc tcctggcctc acgaccctcc ctgaggccag cgatgtgggc 1740 agcatcactc ttgccctccg aacgcttggc agctttgaat ttgaaggcca ctctctgacc 1800 caatttgttc gccactgtgc ggatcatttc ctgaacagtg agcacaagga gatccgcatg 1860 gaggctgccc gcacctgctc ccgcctgctc acaccctcca tccacctcat cagtggccat 1920 gctcatgtgg ttagccagac cgcagtgcaa gtggtggcag atgtgcttag caaactgctc 1980 gtagttggga taacagatcc tgaccctgac attcgctact gtgtcttggc gtccctggac 2040 gagcgctttg atgcacacct ggcccaggcg gagaacttgc aggccttgtt tgtggctctg 2100 aatgaccagg tgtttgagat ccgggagctg gccatctgca ctgtgggccg actcagtagc 2160 atgaaccctg cctttgtcat gcctttcctg cgcaagatgc tcatccagat tttgacagag 2220 ttggagcaca gtgggattgg aagaatcaaa gagcagagtg cccgcatgct ggggcacctg 2280 gtctccaatg ccccccgact catccgcccc tacatggagc ctattctgaa ggcattaatt 2340 ttgaaactga aagatccaga ccctgatcca aacccaggtg tgatcaataa tgtcctggca 2400 acaataggag aattggcaca ggttagtggc ctggaaatga ggaaatgggt tgatgaactt 2460 tttattatca tcatggacat gctccaggat tcctctttgt tggccaaaag gcaggtggct 2520 ctgtggaccc tgggacagtt ggtggccagc actggctatg tagtagagcc ctacaggaag 2580 taccctactt tgcttgaggt gctactgaat tttctgaaga ctgagcagaa ccagggtaca 2640 cgcagagagg ccatccgtgt gttagggctt ttaggggctt tggatcctta caagcacaaa 2700 gtgaacattg gcatgataga ccagtcccgg gatgcctctg ctgtcagcct gtcagaatcc 2760 aagtcaagtc aggattcctc tgactatagc actagtgaaa tgctggtcaa catgggaaac 2820 ttgcctctgg atgagttcta cccagctgtg tccatggtgg ccctgatgcg gatcttccga 2880 gaccagtcac tctctcatca tcacaccatg gttgtccagg ccatcacctt catcttcaag 2940 tccctgggac tcaaatgtgt gcagttcctg ccccaggtca tgcccacgtt ccttaacgtc 3000 attcgagtct gtgatggggc catccgggaa tttttgttcc agcagctggg aatgttggtg 3060 tcctttgtga agagccacat cagaccttat atggatgaaa tagtcaccct catgagagaa 3120 ttctgggtca tgaacacctc aattcagagc acgatcattc ttctcattga gcaaattgtg 3180 gtagctcttg ggggtgaatt taagctctac ctgccccagc tgatcccaca catgctgcgt 3240 gtcttcatgc atgacaacag cccaggccgc attgtctcta tcaagttact ggctgcaatc 3300 cagctgtttg gcgccaacct ggatgactac ctgcatttac tgctgcctcc tattgttaag 3360 ttgtttgatg cccctgaagc tccactgcca tctcgaaagg cagcgctaga gactgtggac 3420 cgcctgacgg agtccctgga tttcactgac tatgcctccc ggatcattca ccctattgtt 3480 cgaacactgg accagagccc agaactgcgc tccacagcca tggacacgct gtcttcactt 3540 gtttttcagc tggggaagaa gtaccaaatt ttcattccaa tggtgaataa agttctggtg 3600 cgacaccgaa tcaatcatca gcgctatgat gtgctcatct gcagaattgt caagggatac 3660 accttgctg atgaagagga ggatcctttg atttaccagc atcggatgct taggagtggc 3720 caaggggatg cattggctag tggaccagtg gaaacaggac ccatgaagaa actgcacgtc 3780 agcaccatca acctccaaaa ggcctggggc gctgccagga gggtctccaa agatgactgg 3840 ctggaatggc tgagacggct gagcctggag ctgctgaagg actcatcatc gccctccctg 3900 cgctcctgct gggccctggc acaggcctac aacccgatgg ccagggatct cttcaatgct 3960 gcatttgtgt cctgctggtc tgaactgaat gaagatcaac aggatgagct catcagaagc 4020 atcgagttgg ccctcacctc acaagacatc gctgaagtca cacagaccct cttaaacttg 4080 gctgaattca tggaacacag tgacaagggc cccctgccac tgagagatga caatggcatt 4140 gttctgctgg gtgagagagc tgccaagtgc cgagcatatg ccaaagcact acactacaaa 4200 gaactggagt tccagaaagg ccccacccct gccattctag aatctctcat cagcattaat 4260 aataagctac agcagccgga ggcagcggcc ggagtgttag aatatgccat gaaacacttt 4320 ggagagctgg agatccaggc tacctggtat gagaaactgc acgagtggga ggatgccctt 4380 gtggcctatg acaagaaaat ggacaccaac aaggacgacc cagagctgat gctgggccgc 4440 atgcgctgcc tcgaggcctt gggggaatgg ggtcaactcc accagcagtg ctgtgaaaag 4500 tggaccctgg ttaatgatga gacccaagcc aagatggccc ggatggctgc tgcagctgca 4560 tggggtttag gtcagtggga cagcatggaa gaatacacct gtatgatccc tcgggacacc 4620 catgatgggg cattttatag agctgtgctg gcactgcatc aggacctctt ctccttggca 4680 caacagtgca ttgacaaggc cagggacctg ctggatgctg aattaactgc gatggcagga 4740 gagagttaca gtcgggcata tggggccatg gtttcttgcc acatgctgtc cgagctggag 4800 gaggttatcc agtacaaact tgtccccgag cgacgagaga tcatccgcca gatctggtgg 4860 gagagactgc agggctgcca gcgtatcgta gaggactggc agaaaatcct tatggtgcgg 4920 tcccttgtgg tcagccctca tgaagacatg agaacctggc tcaagtatgc aagcctgtgc 4980 ggcaagagtg gcaggctggc tcttgctcat aaaactttag tgttgctcct gggagttgat 5040 ccgtctcggc aacttgacca tcctctgcca acagttcacc ctcaggtgac ctatgcctac 5100 atgaaaaaca tgtggaagag tgcccgcaag atcgatgcct tccagcacat gcagcatttt 5160 gtccagacca tgcagcaaca ggcccagcat gccatcgcta ctgaggacca gcagcataag 5220 caggaactgc acaagctcat ggcccgatgc ttcctgaaac ttggagagtg gcagctgaat 5280 ctacagggca tcaatgagag cacaatcccc aaagtgctgc agtactacag cgccgccaca 5340 gagcacgacc gcagctggta caaggcctgg catgcgtggg cagtgatgaa cttcgaagct 5400 gtgctacact acaaacatca gaaccaagcc cgcgatgaga agaagaaact gcgtcatgcc 5460 agcggggcca acatcaccaa cgccaccact gccgccacca cggccgccac tgccaccacc 5520 actgccagca ccgagggcag caacagtgag agcgaggccg agagcaccga gaacagcccc 5580 accccatcgc cgctgcagaa gaaggtcact gaggatctgt ccaaaaccct cctgatgtac 5640 acggtgcctg ccgtccaggg cttcttccgt tccatctcct tgtcacgagg caacaacctc 5700 caggatacac tcagagttct caccttatgg tttgattatg gtcactggcc agatgtcaat 5760 gaggccttag tggagggggt gaaagccatc cagattgata cctggctaca ggttatacct 5820 cagctcattg caagaattga tacgcccaga cccttggtgg gacgtctcat tcaccagctt 5880 ctcacagaca ttggtcggta ccacccccag gccctcatct acccactgac agtggcttct 5940 aagtctacca cgacagcccg gcacaatgca gccaacaaga ttctgaagaa catgtgtgag 6000 cacagcaaca ccctggtcca gcaggccatg atggtgagcg aggagctgat ccgagtggcc 6060 atcctctggc atgagatgtg gcatgaaggc ctggaagagg catctcgttt gtactttggg 6120 gaaaggaacg tgaaaggcat gtttgaggtg ctggagccct tgcatgctat gatggaacgg 6180 ggcccccaga ctctgaagga aacatccttt aatcaggcct atggtcgaga tttaatggag 6240 gcccaagagt ggtgcaggaa gtacatgaaa tcagggaatg tcaaggacct cacccaagcc 6300 tgggacctct attatcatgt gttccgacga atctcaaagc agctgcctca gctcacatcc 6360 ttagagctgc aatatgtttc cccaaaactt ctgatgtgcc gggaccttga attggctgtg 6420 ccaggaacat atgaccccaa ccagccaatc attcgcattc agtccatagc accgtctttg 6480 caagtcatca catccaagca gaggccccgg aaattgacac ttatgggcag caacggacat 6540 gagtttgttt tccttctaaa aggccatgaa gatctgcgcc aggatgagcg tgtgatgcag 6600 ctcttcggcc tggttaacac ccttctggcc aatgacccaa catctcttcg gaaaaacctc 6660 agcatccaga gatacgctgt catcccttta tcgaccaact cgggcctcat tggctgggtt 6720 ccccactgtg acacactgca cgccctcatc cgggactaca gggagaagaa gaagatcctt 6780 ctcaacatcg agcatcgcat catgttgcgg atggctccgg actatgacca cttgactctg 6840 atgcagaagg tggaggtgtt tgagcatgcc gtcaataata cagctgggga cgacctggcc 6900 aagctgctgt ggctgaaaag ccccagctcc gaggtgtggt ttgaccgaag aaccaattat 6960 acccgttctt tagcggtcat gtcaatggtt gggtatattt taggcctggg agatagacac 7020 ccatccaacc tgatgctgga ccgtctgagt gggaagatcc tgcacattga ctttggggac 7080 tgctttgagg ttgctatgac ccgagagaag tttccagaga agattccatt tagactaaca 7140 agaatgttga ccaatgctat ggaggttaca ggcctggatg gcaactacag aatcacatgc 7200 cacacagtga tggaggtgct gcgagagcac aaggacagtg tcatggccgt gctggaagcc 7260 tttgtctatg accccttgct gaactggagg ctgatggaca caaataccaa aggcaacaag 7320 cgatcccgaa cgaggacgga ttcctactct gctggccagt cagtcgaaat tttggacggt 7380 gtggaacttg gagagccagc ccataagaaa acggggacca cagtgccaga atctattcat 7440 tctttcattg gagacggttt ggtgaaacca gaggccctaa ataagaaagc tatccagatt 7500 attaacaggg ttcgagataa gctcactggt cgggacttct ctcatgatga cactttggat 7560 gttccaacgc aagttgagct gctcatcaaa caagcgacat cccatgaaaa cctctgccag 7620 tgctatattg gctggtgccc tttctggtaa 7650 <210> 2 <211> 2549 <212> PRT <213> Homo sapiens_mTOR <400> 2 Met Leu Gly Thr Gly Pro Ala Ala Ala Thr Thr Ala Ala Thr Thr Ser   1 5 10 15 Ser Asn Val Ser Val Leu Gln Gln Phe Ala Ser Gly Leu Lys Ser Arg              20 25 30 Asn Glu Glu Thr Arg Ala Lys Ala Ala Lys Glu Leu Gln His Tyr Val          35 40 45 Thr Met Glu Leu Arg Glu Met Ser Gln Glu Glu Ser Thr Arg Phe Tyr      50 55 60 Asp Gln Leu Asn His His Ile Phe Glu Leu Val Ser Ser Ser Asp Ala  65 70 75 80 Asn Glu Arg Lys Gly Gly Ile Leu Ala Ile Ala Ser Leu Ile Gly Val                  85 90 95 Glu Gly Gly Asn Ala Thr Arg Ile Gly Arg Phe Ala Asn Tyr Leu Arg             100 105 110 Asn Leu Leu Pro Ser Asn Pro Val Val Met Glu Met Ala Ser Lys         115 120 125 Ala Ile Gly Arg Leu Ala Met Ala Gly Asp Thr Phe Thr Ala Glu Tyr     130 135 140 Val Glu Phe Glu Val Lys Arg Ala Leu Glu Trp Leu Gly Ala Asp Arg 145 150 155 160 Asn Glu Gly Arg Arg His Ala Ala Val Leu Val Leu Arg Glu Leu Ala                 165 170 175 Ile Ser Val Pro Thr Phe Phe Phe Gln Gln Val Gln Pro Phe Phe Asp             180 185 190 Asn Ile Phe Val Ala Val Trp Asp Pro Lys Gln Ala Ile Arg Glu Gly         195 200 205 Ala Val Ala Ala Leu Arg Ala Cys Leu Ile Leu Thr Thr Gln Arg Glu     210 215 220 Pro Lys Glu Met Gln Lys Pro Gln Trp Tyr Arg His Thr Phe Glu Glu 225 230 235 240 Ala Glu Lys Gly Phe Asp Glu Thr Leu Ala Lys Glu Lys Gly Met Asn                 245 250 255 Arg Asp Asp Arg Ile His Gly Ala Leu Leu Ile Leu Asn Glu Leu Val             260 265 270 Arg Ile Ser Ser Met Glu Gly Glu Arg Leu Arg Glu Glu Met Glu Glu         275 280 285 Ile Thr Gln Gln Gln Leu Val His Asp Lys Tyr Cys Lys Asp Leu Met     290 295 300 Gly Phe Gly Thr Lys Pro Arg His Ile Thr Pro Phe Thr Ser Phe Gln 305 310 315 320 Ala Val Gln Pro Gln Gln Ser Asn Ala Leu Val Gly Leu Leu Gly Tyr                 325 330 335 Ser Ser His Gln Gly Leu Met Gly Phe Gly Thr Ser Ser Ser Ser Ala             340 345 350 Lys Ser Thr Leu Val Glu Ser Arg Cys Cys Arg Asp Leu Met Glu Glu         355 360 365 Lys Phe Asp Gln Val Cys Gln Trp Val Leu Lys Cys Arg Asn Ser Lys     370 375 380 Asn Ser Leu Ile Gln Met Thr Ile Leu Asn Leu Leu Pro Arg Leu Ala 385 390 395 400 Ala Phe Arg Pro Ser Ala Phe Thr Asp Thr Gln Tyr Leu Gln Asp Thr                 405 410 415 Met Asn His Val Leu Ser Cys Val Lys Lys Glu Lys Glu Arg Thr Ala             420 425 430 Ala Phe Gln Ala Leu Gly Leu Leu Ser Val Ala Val Arg Ser Glu Phe         435 440 445 Lys Val Tyr Leu Pro Arg Val Leu Asp Ile Ile Arg Ala Ala Leu Pro     450 455 460 Pro Lys Asp Phe Ala His Lys Arg Gln Lys Ala Met Gln Val Asp Ala 465 470 475 480 Thr Val Phe Thr Cys Ile Ser Met Leu Ala Arg Ala Met Gly Pro Gly                 485 490 495 Ile Gln Gln Asp Ile Lys Glu Leu Leu Glu Pro Met Leu Ala Val Gly             500 505 510 Leu Ser Pro Ala Leu Thr Ala Val Leu Tyr Asp Leu Ser Arg Gln Ile         515 520 525 Pro Gln Leu Lys Lys Asp Ile Gln Asp Gly Leu Leu Lys Met Leu Ser     530 535 540 Leu Val Leu Met His Lys Pro Leu Arg His Pro Gly Met Pro Lys Gly 545 550 555 560 Leu Ala His Gln Leu Ala Ser Pro Gly Leu Thr Thr Leu Pro Glu Ala                 565 570 575 Ser Asp Val Gly Ser Ile Thr Leu Ala Leu Arg Thr Leu Gly Ser Phe             580 585 590 Glu Phe Glu Gly His Ser Leu Thr Gln Phe Val Arg His Cys Ala Asp         595 600 605 His Phe Leu Asn Ser Glu His Lys Glu Ile Arg Met Glu Ala Ala Arg     610 615 620 Thr Cys Ser Arg Leu Leu Thr Pro Ser Ile His Leu Ile Ser Gly His 625 630 635 640 Ala His Val Val Ser Gln Thr Ala Val Gln Val Val Ala Asp Val Leu                 645 650 655 Ser Lys Leu Leu Val Val Gly Ile Thr Asp Pro Asp Pro Asp Ile Arg             660 665 670 Tyr Cys Val Leu Ala Ser Leu Asp Glu Arg Phe Asp Ala His Leu Ala         675 680 685 Gln Ala Glu Asn Leu Gln Ala Leu Phe Val Ala Leu Asn Asp Gln Val     690 695 700 Phe Glu Ile Arg Glu Leu Ala Ile Cys Thr Val Gly Arg Leu Ser Ser 705 710 715 720 Met Asn Pro Ala Phe Val Met Pro Phe Leu Arg Lys Met Leu Ile Gln                 725 730 735 Ile Leu Thr Glu Leu Glu His Ser Gly Ile Gly Arg Ile Lys Glu Gln             740 745 750 Ser Ala Arg Met Leu Gly His Leu Val Ser Asn Ala Pro Arg Leu Ile         755 760 765 Arg Pro Tyr Met Glu Pro Ile Leu Lys Ala Leu Ile Leu Lys Leu Lys     770 775 780 Asp Pro Asp Pro Asp Pro Asn Pro Gly Val Ile Asn Asn Val Leu Ala 785 790 795 800 Thr Ile Gly Glu Leu Ala Gln Val Ser Gly Leu Glu Met Arg Lys Trp                 805 810 815 Val Asp Glu Leu Phe Ile Ile Ile Met Asp Met Leu Gln Asp Ser Ser             820 825 830 Leu Leu Ala Lys Arg Gln Val Ala Leu Trp Thr Leu Gly Gln Leu Val         835 840 845 Ala Ser Thr Gly Tyr Val Val Glu Pro Tyr Arg Lys Tyr Pro Thr Leu     850 855 860 Leu Glu Val Leu Leu Asn Phe Leu Lys Thr Glu Gln Asn Gln Gly Thr 865 870 875 880 Arg Arg Glu Ala Ile Arg Val Leu Gly Leu Leu Gly Ala Leu Asp Pro                 885 890 895 Tyr Lys His Lys Val Asn Ile Gly Met Ile Asp Gln Ser Arg Asp Ala             900 905 910 Ser Ala Val Ser Leu Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser As Ser         915 920 925 Tyr Ser Thr Ser Glu Met Leu Val Asn Met Gly Asn Leu Pro Leu Asp     930 935 940 Glu Phe Tyr Pro Ala Val Ser Met Val Ala Leu Met Arg Ile Phe Arg 945 950 955 960 Asp Gln Ser Leu Ser His His His Thr Met Val Val Gln Ala Ile Thr                 965 970 975 Phe Ile Phe Lys Ser Leu Gly Leu Lys Cys Val Gln Phe Leu Pro Gln             980 985 990 Val Met Pro Thr Phe Leu Asn Val Ile Arg Val Cys Asp Gly Ala Ile         995 1000 1005 Arg Glu Phe Leu Phe Gln Gln Leu Gly Met Leu Val Ser Phe Val Lys    1010 1015 1020 Ser His Ile Arg Pro Tyr Met Asp Glu Ile Val Thr Leu Met Arg Glu 1025 1030 1035 1040 Phe Trp Val Met Asn Thr Ser Ile Gln Ser Thr Ile Ile Leu Leu Ile                1045 1050 1055 Glu Gln Ile Val Val Ala Leu Gly Gly Glu Phe Lys Leu Tyr Leu Pro            1060 1065 1070 Gln Leu Ile Pro His Met Leu Arg Val Phe Met His Asp Asn Ser Pro        1075 1080 1085 Gly Arg Ile Val Ser Ile Lys Leu    1090 1095 1100 Ala Asn Leu Asp Asp Tyr Leu His Leu Leu Leu Pro Pro Ile Val Lys 1105 1110 1115 1120 Leu Phe Asp Ala Pro Glu Ala Pro Leu Pro Ser Arg Lys Ala Ala Leu                1125 1130 1135 Glu Thr Val Asp Arg Leu Thr Glu Ser Leu Asp Phe Thr Asp Tyr Ala            1140 1145 1150 Ser Arg Ile Ile His Pro Ile Val Arg Thr Leu Asp Gln Ser Pro Glu        1155 1160 1165 Leu Arg Ser Thr Ala Met Asp Thr Leu Ser Ser Leu Val Phe Gln Leu    1170 1175 1180 Gly Lys Lys Tyr Gln Ile Phe Ile Pro Met Val Asn Lys Val Leu Val 1185 1190 1195 1200 Arg His Arg Ile Asn His Gln Arg Tyr Asp Val Leu Ile Cys Arg Ile                1205 1210 1215 Val Lys Gly Tyr Thr Leu Ala Asp Glu Glu Asp Pro Leu Ile Tyr            1220 1225 1230 Gln His Arg Met Leu Arg Ser Gly Gln Gly Asp Ala Leu Ala Ser Gly        1235 1240 1245 Pro Val Glu Thr Gly Pro Met Lys Lys Leu His Val Ser Thr Ile Asn    1250 1255 1260 Leu Gln Lys Ala Trp Gly Ala Ala Arg Arg Val Ser Lys Asp Asp Trp 1265 1270 1275 1280 Leu Glu Trp Leu Arg Arg Leu Ser Leu Glu Leu Leu Lys Asp Ser Ser                1285 1290 1295 Ser Pro Ser Leu Arg Ser Cys Trp Ala Leu Ala Gln Ala Tyr Asn Pro            1300 1305 1310 Met Ala Arg Asp Leu Phe Asn Ala Ala Phe Val Ser Cys Trp Ser Glu        1315 1320 1325 Leu Asn Glu Asp Glu Gln Asp Glu Leu Ile Arg Ser Ile Glu Leu Ala    1330 1335 1340 Leu Thr Ser Gln Asp Ile Ala Glu Val Thr Gln Thr Leu Leu Asn Leu 1345 1350 1355 1360 Ala Glu Phe Met Glu His Ser Asp Lys Gly Pro Leu Pro Leu Arg Asp                1365 1370 1375 Asp Asn Gly Ile Val Leu Leu Gly Glu Arg Ala Ala Lys Cys Arg Ala            1380 1385 1390 Tyr Ala Lys Ala Leu His Tyr Lys Glu Leu Glu Phe Gln Lys Gly Pro        1395 1400 1405 Thr Pro Ala Ile Leu Glu Ser Leu Ile Ser Ile Asn Asn Lys Leu Gln    1410 1415 1420 Gln Pro Glu Ala Ala Gly Val Leu Glu Tyr Ala Met Lys His Phe 1425 1430 1435 1440 Gly Glu Leu Glu Ile Gln Ala Thr Trp Tyr Glu Lys Leu His Glu Trp                1445 1450 1455 Glu Asp Ala Leu Val Ala Tyr Asp Lys Lys Met Asp Thr Asn Lys Asp            1460 1465 1470 Asp Pro Glu Leu Met Leu Gly Arg Met Cys Leu Glu Ala Leu Gly        1475 1480 1485 Glu Trp Gly Gln Leu His Gln Gln Cys Cys Glu Lys Trp Thr Leu Val    1490 1495 1500 Asn Asp Glu Thr Gln Ala Lys Met Ala Arg Met Ala Ala Ala Ala Ala 1505 1510 1515 1520 Trp Gly Leu Gly Gln Trp Asp Ser Met Glu Glu Tyr Thr Cys Met Ile                1525 1530 1535 Pro Arg Asp Thr His Asp Gly Ala Phe Tyr Arg Ala Val Leu Ala Leu            1540 1545 1550 His Gln Asp Leu Phe Ser Leu Ala Gln Gln Cys Ile Asp Lys Ala Arg        1555 1560 1565 Asp Leu Leu Asp Ala Glu Leu Thr Ala Met Ala Gly Glu Ser Tyr Ser    1570 1575 1580 Arg Ala Tyr Gly Ala Met Val Ser Cys His Met Leu Ser Glu Leu Glu 1585 1590 1595 1600 Glu Val Ile Gln Tyr Lys Leu Val Pro Glu Arg Arg Glu Ile Ile Arg                1605 1610 1615 Gln Ile Trp Trp Glu Arg Leu Gln Gly Cys Gln Arg Ile Val Glu Asp            1620 1625 1630 Trp Gln Lys Ile Leu Met Val Arg Ser Leu Val Val Ser Pro His Glu        1635 1640 1645 Asp Met Arg Thr Trp Leu Lys Tyr Ala Ser Leu Cys Gly Lys Ser Gly    1650 1655 1660 Arg Leu Ala Leu Ala His Lys Thr Leu Val Leu Leu Leu Gly Val Asp 1665 1670 1675 1680 Pro Ser Arg Gln Leu Asp His Pro Leu Pro Thr Val His Pro Gln Val                1685 1690 1695 Thr Tyr Ala Tyr Met Lys Asn Met Trp Lys Ser Ala Arg Lys Ile Asp            1700 1705 1710 Ala Phe Gln His Met Gln His Phe Val Gln Thr Met Gln Gln Gln Ala        1715 1720 1725 Gln His Ala Ile Ala Thr Glu Asp Gln Gln His Lys Gln Glu Leu His    1730 1735 1740 Lys Leu Met Ala Arg Cys Phe Leu Lys Leu Gly Glu Trp Gln Leu Asn 1745 1750 1755 1760 Leu Gln Gly Ile Asn Glu Ser Thr Ile Pro Lys Val Leu Gln Tyr Tyr                1765 1770 1775 Ser Ala Ala Thr Glu His Asp Arg Ser Trp Tyr Lys Ala Trp His Ala            1780 1785 1790 Trp Ala Val Met Asn Phe Glu Ala Val Leu His Tyr Lys His Gln Asn        1795 1800 1805 Gln Ala Arg Asp Glu Lys Lys Lys Leu Arg His Ala Ser Gly Ala Asn    1810 1815 1820 Ile Thr Asn Ala Thr Thr Ala Thr Thr 1825 1830 1835 1840 Thr Ala Ser Thr Glu Gly Ser Ser                1845 1850 1855 Glu Asn Ser Pro Thr Pro Ser Pro Leu Gln Lys Lys Val Thr Glu Asp            1860 1865 1870 Leu Ser Lys Thr Leu Leu Met Tyr Thr Val Ala Val Gln Gly Phe        1875 1880 1885 Phe Arg Ser Ser Leu Ser Arg Gly Asn Asn Leu Gln Asp Thr Leu    1890 1895 1900 Arg Val Leu Thr Leu Trp Phe Asp Tyr Gly His Trp Pro Asp Val Asn 1905 1910 1915 1920 Glu Ala Leu Val Glu Gly Val Lys Ala Ile Gln Ile Asp Thr Trp Leu                1925 1930 1935 Gln Val Ile Pro Gln Leu Ile Ala Arg Ile Asp Thr Pro Arg Pro Leu            1940 1945 1950 Val Gly Arg Leu Ile His Gln Leu Leu Thr Asp Ile Gly Arg Tyr His        1955 1960 1965 Pro Gln Ala Leu Ile Tyr Pro Leu Thr Val Ala Ser Lys Ser Thr Thr    1970 1975 1980 Thr Ala Arg His Asn Ala Ala Asn Lys Ile Leu Lys Asn Met Cys Glu 1985 1990 1995 2000 His Ser Asn Thr Leu Val Gln Gln Ala Met Met Val Ser Glu Glu Leu                2005 2010 2015 Ile Arg Val Ala Ile Leu Trp His Glu Met Trp His Glu Gly Leu Glu            2020 2025 2030 Glu Ala Ser Arg Leu Tyr Phe Gly Glu Arg Asn Val Lys Gly Met Phe        2035 2040 2045 Glu Val Leu Glu Pro Leu His Ala Met Met Glu Arg Gly Pro Gln Thr    2050 2055 2060 Leu Lys Glu Thr Ser Phe Asn Gln Ala Tyr Gly Arg Asp Leu Met Glu 2065 2070 2075 2080 Ala Gln Glu Trp Cys Arg Lys Tyr Met Lys Ser Gly Asn Val Lys Asp                2085 2090 2095 Leu Thr Gln Ala Trp Asp Leu Tyr Tyr His Val Phe Arg Arg Ile Ser            2100 2105 2110 Lys Gln Leu Pro Gln Leu Thr Ser Leu Glu Leu Gln Tyr Val Ser Pro        2115 2120 2125 Lys Leu Leu Met Cys Arg Asp Leu Glu Leu Ala Val Pro Gly Thr Tyr    2130 2135 2140 Asp Pro Asn Gln Pro Ile Ile Arg Ile Gln Ser Ile Ala Pro Ser Leu 2145 2150 2155 2160 Gln Val Ile Thr Ser Lys Gln Arg Pro Arg Lys Leu Thr Leu Met Gly                2165 2170 2175 Ser Asn Gly His Glu Phe Val Phe Leu Leu Lys Gly His Glu Asp Leu            2180 2185 2190 Arg Gln Asp Glu Arg Val Met Gln Leu Phe Gly Leu Val Asn Thr Leu        2195 2200 2205 Leu Ala Asn Asp Pro Thr Ser Leu Arg Lys Asn Leu Ser Ile Gln Arg    2210 2215 2220 Tyr Ala Val Ile Pro Leu Ser Thr Asn Ser Gly Leu Ile Gly Trp Val 2225 2230 2235 2240 Pro His Cys Asp Thr Leu His Ala Leu Ile Arg Asp Tyr Arg Glu Lys                2245 2250 2255 Lys Lys Ile Leu Leu Asn Ile Glu His Arg Ile Met Leu Arg Met Ala            2260 2265 2270 Pro Asp Tyr Asp His Leu Thr Leu Met Gln Lys Val Glu Val Phe Glu        2275 2280 2285 His Ala Val Asn Asn Thr Ala Gly Asp Asp Leu Ala Lys Leu Leu Trp    2290 2295 2300 Leu Lys Ser Pro Ser Ser Glu Val Trp Phe Asp Arg Arg Thr Asn Tyr 2305 2310 2315 2320 Thr Arg Ser Leu Ala Val Met Ser Met Val Gly Tyr Ile Leu Gly Leu                2325 2330 2335 Gly Asp Arg His Pro Ser Asn Leu Met Leu Asp Arg Leu Ser Gly Lys            2340 2345 2350 Ile Leu His Ile Asp Phe Gly Asp Cys Phe Glu Val Ala Met Thr Arg        2355 2360 2365 Glu Lys Phe Pro Glu Lys Ile Pro Phe Arg Leu Thr Arg Met Leu Thr    2370 2375 2380 Asn A Met Glu Val Thr Gly Leu Asp Gly Asn Tyr Arg Ile Thr Cys 2385 2390 2395 2400 His Thr Val Met Glu Val Leu Arg Glu His Lys Asp Ser Val Met Ala                2405 2410 2415 Val Leu Glu Ala Phe Val Tyr Asp Pro Leu Leu Asn Trp Arg Leu Met            2420 2425 2430 Asp Thr Asn Thr Lys Gly Asn Lys Arg Ser Ser Thr Arg Thr Asp Ser        2435 2440 2445 Tyr Ser Ala Gly Gln Ser Val Glu Ile Leu Asp Gly Val Glu Leu Gly    2450 2455 2460 Glu Pro Ala His Lys Lys Thr Gly Thr Thr Val Pro Glu Ser Ile His 2465 2470 2475 2480 Ser Phe Ile Gly Asp Gly Leu Val Lys Pro Glu Ala Leu Asn Lys Lys                2485 2490 2495 Ala Ile Gln Ile Ile Asn Arg Val Arg Asp Lys Leu Thr Gly Arg Asp            2500 2505 2510 Phe Ser His Asp Asp Thr Leu Asp Val Pro Thr Gln Val Glu Leu Leu        2515 2520 2525 Ile Lys Gln Ala Thr Ser His Glu Asn Leu Cys Gln Cys Tyr Ile Gly    2530 2535 2540 Trp Cys Pro Phe Trp 2545 <210> 3 <211> 7650 <212> DNA <213> mouse_mTOR <400> 3 atgcttggga cgggtcctgc cgtggccacc gccagtgccg ccacatctag caacgtgagc 60 gtcctgcagc agttcgccag tggactgaag agccggaatg aggagaccag ggccaaagca 120 gccaaggagc tccagcacta cgtcaccatg gagcttcgag agatgagtca ggaggagtct 180 actcgcttct atgaccagct gaaccatcac atttttgaac tggtttccag ctcagatgcc 240 aatgagagga agggtggcat cttggccatc gccagcctca taggagtgga aggtgggaat 300 tccaccagaa ttggcagatt tgccaactac cttcgaaacc tcctcccctc aagcgatcca 360 gttgtcatgg aaatggcgtc caaggccatt ggccgcctgg cgatggcagg ggacactttc 420 actgctgaat atgtggagtt tgaagtgaag cgagccttgg agtggctggg tgctgaccga 480 aatgagggcc ggagacatgc cgctgtcctc gttctccgtg agctggccat cagtgtcccc 540 accttcttct tccagcaagt tcagcccttc tttgacaaca tttttgtggc tgtgtgggac 600 cccaagcagg ccatccggga aggcgctgta gcggcccttc gtgcctgtct gattctcacc 660 acgcagcggg aaccaaagga aatgcagaag cctcagtggt accggcacac atttgaagaa 720 gcagagaaag gttttgatga gaccctggcc aaagagaagg gtatgaatcg agatgatcga 780 atccacggag ccttgctgat cctcaacgag ctagttcgta tcagcagcat ggagggagag 840 cgtctgagag aagagatgga ggagatcacc cagcagcagc tggtgcatga caagtactgc 900 aaagacctca tgggcttcgg gaccaagcct cggcacatca cgcccttcac cagtttccag 960 gctgtgcagc cccagcagcc gaacgccttg gtgggactgc tggggtacag ctcccctcaa 1020 ggcctgatgg gatttgggac gtcccccagc cctgccaagt ccactctggt ggaaagccgc 1080 tgttgcagag acttgatgga agagaaattt gatcaggtgt gccagtgggt gctgaagtgc 1140 aggagcagca agaactcgct gatccagatg acaatcctta acctgctgcc ccgcctggct 1200 gcattccgac cgtccgcctt cacagatacc cagtacctcc aggacaccat gaaccatgtc 1260 ctgagctgtg tcaagaagga gaaggaacgg actgcggcgt tccaggccct ggggctgctt 1320 tctgtggccg tgaggtcgga gtttaaggtc tacttgcccc gtgtacttga catcatccga 1380 gcagcgcttc ctccaaagga ctttgcccac aagaggcaga aaaccgtgca ggtggatgcc 1440 accgtattca cgtgcatcag catgttggca cgagcaatgg ggccgggcat ccagcaggac 1500 atcaaggagc tgctggagcc catgttggca gtgggcctga gccccgcgct cactgctgtg 1560 ctctatgacc tgagccggca gattccacag ctgaagaaag atattcagga cggccttctg 1620 aagatgctgt ccctggtcct tatgcacaaa cccctccggc acccaggcat gcccaaaggc 1680 ctggctcacc agctggcttc ccctggtctc accaccctcc ctgaggccag cgacgtggcc 1740 agcatcactc ttgcccttcg aacccttggc agctttgaat ttgaaggcca ctctctgacc 1800 cagttcgtcc gacactgcgc agatcacttc ctgaacagcg agcacaagga gatccgcatg 1860 gaagctgctc gcacctgctc ccgcctgctc acaccctcca tccacctcat cagcggccat 1920 gcccacgtgg ttagccagac tgcagtgcag gtggtggcag atgtgctcag caagctgctt 1980 gtggttggca taacagatcc tgaccctgat atccgctact gtgtcttggc atccctggac 2040 gagcgctttg atgcccacct ggcccaggca gaaaacttac aagctctgtt tgtggctctg 2100 aatgaccagg tctttgagat ccgcgagctg gccatctgca ctgtgggccg actaagcagc 2160 atgaacccag ccttcgtcat gcctttcctg cgcaagatgc tcatccagat cctgacagag 2220 ctggagcaca gcggcattgg gagaatcaag gagcagagcg cccgcatgct ggggcacctg 2280 gtgtccaacg ccccccggct catccgcccc tacatggagc ctatcctgaa ggctttaatt 2340 ttgaaactga aagatccaga ccctgaccca aacccgggcg tgatcaataa cgtgttggcc 2400 actataggag aactggctca ggtgagcggc ctggaaatgc ggaagtgggt ggacgagctc 2460 tttatcatca tcatggacat gctgcaggac tcctccctgc tggccaaaag gcaggtggct 2520 ttgtggaccc tgggacagtt ggtggccagc actggctatg tggtggagcc ctacaggaag 2580 taccccactt tgcttgaagt gctgctgaat ttcctgaaga cggagcagaa ccagggcact 2640 cggagagagg ctatccgagt gttggggctc cttggggctt tggatcccta caagcacaaa 2700 gtgaacatcg gcatgatcga ccagtcccgg gacgcttccg ctgtcagcct gtcagagtcc 2760 aagtcaagtc aggattcctc tgactacagc accagtgaaa tgctggtcaa catgggaaac 2820 ctgcccctgg acgagttcta ccccgctgtg tccatggtgg ccttgatgcg gatcttccga 2880 gatcaatccc tctctcacca ccacaccatg gtggtgcagg ccatcacctt catcttcaag 2940 tccctggggc tcaagtgtgt gcagttcctg ccccaggtca tgcccacatt ccttaatgtc 3000 atccgagtct gtgatggggc catccgggaa tttctgttcc agcagctggg gatgctggtg 3060 tcctttgtga agagccacat ccgtccctac atggatgaaa tagtcactct catgagagag 3120 ttttgggtca tgaacacgtc aatccagagc acaatcattc ttctcattga gcagattgtg 3180 gtggctctcg gaggggaatt taagctttat ctgccccagt tgatcccaca catgctgcgg 3240 gtcttcatgc atgacaacag ccaaggccga atcgtctcca tcaagctgtt agccgcgatc 3300 cagctgtttg gcgccaacct ggatgactat ctgcacttgt tgttgcctcc gattgtgaaa 3360 ttgtttgatg cccctgaagt cccgctgcca tcaagaaagg cagcgctgga gacggtggac 3420 cgcctgacag agtccctaga cttcactgac tacgcctccc gcatcattca cccaatagtt 3480 cgtacgctag accagagccc agagctgcgc tccacagcca tggacactct gtcttcgctt 3540 gtctttcaac tggggaagaa gtaccagatc ttcattccaa tggtgaataa agtcctcgtg 3600 cgacaccgga tcaaccacca gcgctatgat gtgcttatct gcagaatcgt caaggggtac 3660 acacttgctg atgaggaaga agaccctttg atttaccagc atcgaatgct aaggagcagc 3720 cagggagatg ccctggccag tggaccagtt gagacaggac ccatgaagaa actgcatgtc 3780 agcaccatca acctccaaaa ggcctgggga gctgccagaa gggtctccaa ggacgactgg 3840 ctggagtggc tgaggcgctt gagtctggag cttctgaagg actcctcatc gccctccctg 3900 cgctcatgct gggccctggc gcaggcctac aaccccatgg ccagggatct cttcaatgct 3960 gcctttgtgt cctgctggtc tgagctgaat gaagaccagc aagatgagct catcaggagt 4020 attgagttgg ctctcacttc tcaagacatt gctgaagtca cacaaaccct cctgaacttg 4080 gctgagttca tggaacacag tgacaagggc cccctgccgc tgagagatga caatggcatc 4140 gtgctcctgg gtgagagagc tgccaagtgc cgggcatatg ccaaagcact gcactacaaa 4200 gaactggagt tccagaaagg gcccacgcct gccatacttg agtccctcat cagcattaac 4260 aacaagctcc agcagcctga ggcagcttct ggggtgttgg aatacgccat gaaacacttc 4320 ggagagctgg agatccaggc cacctggtat gagaagctgc atgagtggga ggatgctctc 4380 gtggcctacg acaagaagat ggacacaaac aaggaagacc cggagctgat gctgggccga 4440 atgcgctgcc tcgaggcctt gggggaatgg ggccagcttc atcagcagtg ctgtgaaaag 4500 tggactctgg ttaatgatga gacccaggct aagatggccc ggatggctgc tgctgcagcg 4560 tggggtttag gtcagtggga cagcatggag gagtacacct gcatgatccc acgggacacc 4620 cacgatggag ccttttacag ggcagtgttg gctctacatc aggatctctt ctccttggcc 4680 cagcagtgca ttgacaaggc cagggacctg ctggatgcag agctgactgc catggcagga 4740 gagagctaca gccgagccta tggggccatg gtttcttgcc acatgctgtc cgagctggaa 4800 gaggttatcc agtacaaact tgtccctgag cgtcgggaga tcatccggca gatctggtgg 4860 gagagactgc agggctgcca gcgtattgtt gaggactggc agaaaatcct catggtccgg 4920 tcccttgtgg tcagccctca tgaggacatg agaacctggc tcaagtacgc aagcctgtgt 4980 ggcaagagtg gcagactggc tcttgctcat aaaaccttag tgttgctctt gggagttgat 5040 ccatctcggc aacttgacca tcctctgcca accgctcacc ctcaagtgac ctatgcctac 5100 atgaagaaca tgtggaaaag tgctcggaag attgacgcct tccagcacat gcaacacttt 5160 gtgcagacca tgcagcagca ggcccagcat gccatcgcca cagaggacca gcagcacaag 5220 caggagctgc ataagctcat ggccaggtgt tttctgaaac ttggggagtg gcagctgaac 5280 ctccagggca tcaacgagag caccatcccc aaggtgctac agtactacaga tgccgccaca 5340 gagcatgacc gcagctggta caaggcttgg catgcatggg cagtgatgaa cttcgaagca 5400 gtgctacact acaaacatca gaaccaagcc cgtgatgaga agaagaagct gcgtcatgcc 5460 agcggggcca acatcaccaa tgccaccact gcagccacca ctgcagcctc tgctgctgct 5520 gccaccagca cagagggcag caacagtgag agtgaagctg agagcaatga gaacagcccc 5580 accccgtccc ctctgcagaa gaaggtcact gaggatttat ccaaaactct cttgttgtac 5640 actgtccctg ctgttcaagg cttcttccgt tctatctcct tgtcaagagg caacaacctc 5700 caggatacac taagagtcct caccctgtgg tttgattatg gtcactggcc agatgtcaat 5760 gaagccttgg tggaaggggt gaaggccata cagattgaca cttggttaca ggttatacct 5820 cagctcattg caagaattga cacacccaga cccttggtgg gccggctcat tcaccagctt 5880 ctcacagata ttggtcggta ccacccacag gccctcatct accccctgac ggtggcttct 5940 aagtctacca ccacagcccg tcacaatgca gccaacaaga tcttgaagaa catgtgtgaa 6000 cacagcaaca cgctggtcca gcaggccatg atggtgagtg aagagctgat tcgggtagcc 6060 atcctctggc atgagatgtg gcatgaaggc ctggaagagg catctcgctt gtactttggg 6120 gagaggaacg tgaaaggcat gtttgaggtg ctggagcccc tgcatgctat gatggaacgg 6180 ggtccccaga ctctgaagga aacatccttt aatcaggcat atggccgaga tttaatggag 6240 gcacaagaat ggtgtcgaaa gtacatgaag tcggggaacg tcaaggacct cacgcaagcc 6300 tgggacctct actatcacgt gttcagacgg atctcaaagc agctacccca gctcacatcc 6360 ctggagctgc agtatgtgtc ccccaaactt ctgatgtgcc gagaccttga gttggctgtg 6420 ccaggaacat acgaccccaa ccagccaatc attcgcattc aatccatagc cccgtctttg 6480 caagtcatca catccaagca gaggcctcgg aagctgactc tgatgggcag caatgggcat 6540 gagtttgttt tcctcctgaa aggccatgaa gatctgcggc aggatgaacg agtgatgcag 6600 ctctttggcc tggtgaacac actcctagcc aatgacccca cttctcttcg caagaacctc 6660 agcatccaga gatacgctgt catccctctg tccaccaact cgggcctcat tggctgggtg 6720 ccccactgtg acacactgca tgccctcatc cgggactaca gagagaagaa gaagatcctg 6780 ttgaacatcg agcatcgcat catgctgcgg atggctcctg actatgacca cctgacgttg 6840 atgcagaagg tagaggtgtt tgagcatgct gtcaacaaca cagctgggga cgacctggcc 6900 aagctactgt ggctaaaaag ccccagctcg gaggtgtggt ttgaccgaag aaccaactat 6960 acccgctccc tggccgttat gtcgatggtc ggatacattt taggccttgg agacaggcac 7020 ccatccaatc tgatgctgga ccggctgagt gggaagatcc tgcacattga ctttggggac 7080 tgctttgagg tcgctatgac cagagagaaa tttccagaaa agattccatt tagactaaca 7140 agaatgttga ccaatgctat ggaggttacg ggtctggatg gcaactacag aaccacatgc 7200 cacaccgtga tggaagtgct ccgggaacac aaggacagtg tcatggctgt gctggaagcc 7260 tttgtctatg acccactgct caactggagg ctgatggaca caaataccaa aggcaataag 7320 cggtcccgga caaggacaga ctcctactct gccggccagt cagtagaaat tttggacggt 7380 gtagaacttg gagaaccagc ccataagaaa gcagggacca ctgtgccaga atccatccat 7440 tcattcattg gagacggttt ggtgaaacca gaagccttaa acaagaaagc tattcagatt 7500 attaacaggg ttcgagataa gctcactggt cgggatttct ctcatgatga cactttggat 7560 gttccaccc aagtggagct gcttatcaag caggcaacat ctcacgagaa cctctgccag 7620 tgctacattg gctggtgtcc cttctggtaa 7650 <210> 4 <211> 2549 <212> PRT <213> mouse_mTOR <400> 4 Met Leu Gly Thr Gly Pro Ala Val Ala Thr Ala Ser Ala Ala Thr Ser   1 5 10 15 Ser Asn Val Ser Val Leu Gln Gln Phe Ala Ser Gly Leu Lys Ser Arg              20 25 30 Asn Glu Glu Thr Arg Ala Lys Ala Ala Lys Glu Leu Gln His Tyr Val          35 40 45 Thr Met Glu Leu Arg Glu Met Ser Gln Glu Glu Ser Thr Arg Phe Tyr      50 55 60 Asp Gln Leu Asn His His Ile Phe Glu Leu Val Ser Ser Ser Asp Ala  65 70 75 80 Asn Glu Arg Lys Gly Gly Ile Leu Ala Ile Ala Ser Leu Ile Gly Val                  85 90 95 Glu Gly Asn Ser Thr Arg Ile Gly Arg Phe Ala Asn Tyr Leu Arg             100 105 110 Asn Leu Leu Pro Ser Ser Asp Pro Val Val Met Glu Met Ala Ser Lys         115 120 125 Ala Ile Gly Arg Leu Ala Met Ala Gly Asp Thr Phe Thr Ala Glu Tyr     130 135 140 Val Glu Phe Glu Val Lys Arg Ala Leu Glu Trp Leu Gly Ala Asp Arg 145 150 155 160 Asn Glu Gly Arg Arg His Ala Ala Val Leu Val Leu Arg Glu Leu Ala                 165 170 175 Ile Ser Val Pro Thr Phe Phe Phe Gln Gln Val Gln Pro Phe Phe Asp             180 185 190 Asn Ile Phe Val Ala Val Trp Asp Pro Lys Gln Ala Ile Arg Glu Gly         195 200 205 Ala Val Ala Ala Leu Arg Ala Cys Leu Ile Leu Thr Thr Gln Arg Glu     210 215 220 Pro Lys Glu Met Gln Lys Pro Gln Trp Tyr Arg His Thr Phe Glu Glu 225 230 235 240 Ala Glu Lys Gly Phe Asp Glu Thr Leu Ala Lys Glu Lys Gly Met Asn                 245 250 255 Arg Asp Asp Arg Ile His Gly Ala Leu Leu Ile Leu Asn Glu Leu Val             260 265 270 Arg Ile Ser Ser Met Glu Gly Glu Arg Leu Arg Glu Glu Met Glu Glu         275 280 285 Ile Thr Gln Gln Gln Leu Val His Asp Lys Tyr Cys Lys Asp Leu Met     290 295 300 Gly Phe Gly Thr Lys Pro Arg His Ile Thr Pro Phe Thr Ser Phe Gln 305 310 315 320 Ala Val Gln Pro Gln Gln Pro Asn Ala Leu Val Gly Leu Leu Gly Tyr                 325 330 335 Ser Ser Pro Gln Gly Leu Met Gly Phe Gly Thr Ser Ser Ser Ser Ala             340 345 350 Lys Ser Thr Leu Val Glu Ser Arg Cys Cys Arg Asp Leu Met Glu Glu         355 360 365 Lys Phe Asp Gln Val Cys Gln Trp Val Leu Lys Cys Arg Ser Ser Lys     370 375 380 Asn Ser Leu Ile Gln Met Thr Ile Leu Asn Leu Leu Pro Arg Leu Ala 385 390 395 400 Ala Phe Arg Pro Ser Ala Phe Thr Asp Thr Gln Tyr Leu Gln Asp Thr                 405 410 415 Met Asn His Val Leu Ser Cys Val Lys Lys Glu Lys Glu Arg Thr Ala             420 425 430 Ala Phe Gln Ala Leu Gly Leu Leu Ser Val Ala Val Arg Ser Glu Phe         435 440 445 Lys Val Tyr Leu Pro Arg Val Leu Asp Ile Ile Arg Ala Ala Leu Pro     450 455 460 Pro Lys Asp Phe Ala His Lys Arg Gln Lys Thr Val Gln Val Asp Ala 465 470 475 480 Thr Val Phe Thr Cys Ile Ser Met Leu Ala Arg Ala Met Gly Pro Gly                 485 490 495 Ile Gln Gln Asp Ile Lys Glu Leu Leu Glu Pro Met Leu Ala Val Gly             500 505 510 Leu Ser Pro Ala Leu Thr Ala Val Leu Tyr Asp Leu Ser Arg Gln Ile         515 520 525 Pro Gln Leu Lys Lys Asp Ile Gln Asp Gly Leu Leu Lys Met Leu Ser     530 535 540 Leu Val Leu Met His Lys Pro Leu Arg His Pro Gly Met Pro Lys Gly 545 550 555 560 Leu Ala His Gln Leu Ala Ser Pro Gly Leu Thr Thr Leu Pro Glu Ala                 565 570 575 Ser Asp Val Ala Ser Ile Thr Leu Ala Leu Arg Thr Leu Gly Ser Phe             580 585 590 Glu Phe Glu Gly His Ser Leu Thr Gln Phe Val Arg His Cys Ala Asp         595 600 605 His Phe Leu Asn Ser Glu His Lys Glu Ile Arg Met Glu Ala Ala Arg     610 615 620 Thr Cys Ser Arg Leu Leu Thr Pro Ser Ile His Leu Ile Ser Gly His 625 630 635 640 Ala His Val Val Ser Gln Thr Ala Val Gln Val Val Ala Asp Val Leu                 645 650 655 Ser Lys Leu Leu Val Val Gly Ile Thr Asp Pro Asp Pro Asp Ile Arg             660 665 670 Tyr Cys Val Leu Ala Ser Leu Asp Glu Arg Phe Asp Ala His Leu Ala         675 680 685 Gln Ala Glu Asn Leu Gln Ala Leu Phe Val Ala Leu Asn Asp Gln Val     690 695 700 Phe Glu Ile Arg Glu Leu Ala Ile Cys Thr Val Gly Arg Leu Ser Ser 705 710 715 720 Met Asn Pro Ala Phe Val Met Pro Phe Leu Arg Lys Met Leu Ile Gln                 725 730 735 Ile Leu Thr Glu Leu Glu His Ser Gly Ile Gly Arg Ile Lys Glu Gln             740 745 750 Ser Ala Arg Met Leu Gly His Leu Val Ser Asn Ala Pro Arg Leu Ile         755 760 765 Arg Pro Tyr Met Glu Pro Ile Leu Lys Ala Leu Ile Leu Lys Leu Lys     770 775 780 Asp Pro Asp Pro Asp Pro Asn Pro Gly Val Ile Asn Asn Val Leu Ala 785 790 795 800 Thr Ile Gly Glu Leu Ala Gln Val Ser Gly Leu Glu Met Arg Lys Trp                 805 810 815 Val Asp Glu Leu Phe Ile Ile Ile Met Asp Met Leu Gln Asp Ser Ser             820 825 830 Leu Leu Ala Lys Arg Gln Val Ala Leu Trp Thr Leu Gly Gln Leu Val         835 840 845 Ala Ser Thr Gly Tyr Val Val Glu Pro Tyr Arg Lys Tyr Pro Thr Leu     850 855 860 Leu Glu Val Leu Leu Asn Phe Leu Lys Thr Glu Gln Asn Gln Gly Thr 865 870 875 880 Arg Arg Glu Ala Ile Arg Val Leu Gly Leu Leu Gly Ala Leu Asp Pro                 885 890 895 Tyr Lys His Lys Val Asn Ile Gly Met Ile Asp Gln Ser Arg Asp Ala             900 905 910 Ser Ala Val Ser Leu Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser As Ser         915 920 925 Tyr Ser Thr Ser Glu Met Leu Val Asn Met Gly Asn Leu Pro Leu Asp     930 935 940 Glu Phe Tyr Pro Ala Val Ser Met Val Ala Leu Met Arg Ile Phe Arg 945 950 955 960 Asp Gln Ser Leu Ser His His His Thr Met Val Val Gln Ala Ile Thr                 965 970 975 Phe Ile Phe Lys Ser Leu Gly Leu Lys Cys Val Gln Phe Leu Pro Gln             980 985 990 Val Met Pro Thr Phe Leu Asn Val Ile Arg Val Cys Asp Gly Ala Ile         995 1000 1005 Arg Glu Phe Leu Phe Gln Gln Leu Gly Met Leu Val Ser Phe Val Lys    1010 1015 1020 Ser His Ile Arg Pro Tyr Met Asp Glu Ile Val Thr Leu Met Arg Glu 1025 1030 1035 1040 Phe Trp Val Met Asn Thr Ser Ile Gln Ser Thr Ile Ile Leu Leu Ile                1045 1050 1055 Glu Gln Ile Val Val Ala Leu Gly Gly Glu Phe Lys Leu Tyr Leu Pro            1060 1065 1070 Gln Leu Ile Pro His Met Leu Arg Val Phe Met His Asp Asn Ser Gln        1075 1080 1085 Gly Arg Ile Val Ser Ile Lys Leu    1090 1095 1100 Ala Asn Leu Asp Asp Tyr Leu His Leu Leu Leu Pro Pro Ile Val Lys 1105 1110 1115 1120 Leu Phe Asp Ala Pro Glu Val Pro Leu Pro Ser Arg Lys Ala Ala Leu                1125 1130 1135 Glu Thr Val Asp Arg Leu Thr Glu Ser Leu Asp Phe Thr Asp Tyr Ala            1140 1145 1150 Ser Arg Ile Ile His Pro Ile Val Arg Thr Leu Asp Gln Ser Pro Glu        1155 1160 1165 Leu Arg Ser Thr Ala Met Asp Thr Leu Ser Ser Leu Val Phe Gln Leu    1170 1175 1180 Gly Lys Lys Tyr Gln Ile Phe Ile Pro Met Val Asn Lys Val Leu Val 1185 1190 1195 1200 Arg His Arg Ile Asn His Gln Arg Tyr Asp Val Leu Ile Cys Arg Ile                1205 1210 1215 Val Lys Gly Tyr Thr Leu Ala Asp Glu Glu Asp Pro Leu Ile Tyr            1220 1225 1230 Gln His Arg Met Leu Arg Ser Ser Gln Gly Asp Ala Leu Ala Ser Gly        1235 1240 1245 Pro Val Glu Thr Gly Pro Met Lys Lys Leu His Val Ser Thr Ile Asn    1250 1255 1260 Leu Gln Lys Ala Trp Gly Ala Ala Arg Arg Val Ser Lys Asp Asp Trp 1265 1270 1275 1280 Leu Glu Trp Leu Arg Arg Leu Ser Leu Glu Leu Leu Lys Asp Ser Ser                1285 1290 1295 Ser Pro Ser Leu Arg Ser Cys Trp Ala Leu Ala Gln Ala Tyr Asn Pro            1300 1305 1310 Met Ala Arg Asp Leu Phe Asn Ala Ala Phe Val Ser Cys Trp Ser Glu        1315 1320 1325 Leu Asn Glu Asp Glu Gln Asp Glu Leu Ile Arg Ser Ile Glu Leu Ala    1330 1335 1340 Leu Thr Ser Gln Asp Ile Ala Glu Val Thr Gln Thr Leu Leu Asn Leu 1345 1350 1355 1360 Ala Glu Phe Met Glu His Ser Asp Lys Gly Pro Leu Pro Leu Arg Asp                1365 1370 1375 Asp Asn Gly Ile Val Leu Leu Gly Glu Arg Ala Ala Lys Cys Arg Ala            1380 1385 1390 Tyr Ala Lys Ala Leu His Tyr Lys Glu Leu Glu Phe Gln Lys Gly Pro        1395 1400 1405 Thr Pro Ala Ile Leu Glu Ser Leu Ile Ser Ile Asn Asn Lys Leu Gln    1410 1415 1420 Gln Pro Glu Ala Ala Ser Gly Val Leu Glu Tyr Ala Met Lys His Phe 1425 1430 1435 1440 Gly Glu Leu Glu Ile Gln Ala Thr Trp Tyr Glu Lys Leu His Glu Trp                1445 1450 1455 Glu Asp Ala Leu Val Ala Tyr Asp Lys Lys Met Asp Thr Asn Lys Glu            1460 1465 1470 Asp Pro Glu Leu Met Leu Gly Arg Met Cys Leu Glu Ala Leu Gly        1475 1480 1485 Glu Trp Gly Gln Leu His Gln Gln Cys Cys Glu Lys Trp Thr Leu Val    1490 1495 1500 Asn Asp Glu Thr Gln Ala Lys Met Ala Arg Met Ala Ala Ala Ala Ala 1505 1510 1515 1520 Trp Gly Leu Gly Gln Trp Asp Ser Met Glu Glu Tyr Thr Cys Met Ile                1525 1530 1535 Pro Arg Asp Thr His Asp Gly Ala Phe Tyr Arg Ala Val Leu Ala Leu            1540 1545 1550 His Gln Asp Leu Phe Ser Leu Ala Gln Gln Cys Ile Asp Lys Ala Arg        1555 1560 1565 Asp Leu Leu Asp Ala Glu Leu Thr Ala Met Ala Gly Glu Ser Tyr Ser    1570 1575 1580 Arg Ala Tyr Gly Ala Met Val Ser Cys His Met Leu Ser Glu Leu Glu 1585 1590 1595 1600 Glu Val Ile Gln Tyr Lys Leu Val Pro Glu Arg Arg Glu Ile Ile Arg                1605 1610 1615 Gln Ile Trp Trp Glu Arg Leu Gln Gly Cys Gln Arg Ile Val Glu Asp            1620 1625 1630 Trp Gln Lys Ile Leu Met Val Arg Ser Leu Val Val Ser Pro His Glu        1635 1640 1645 Asp Met Arg Thr Trp Leu Lys Tyr Ala Ser Leu Cys Gly Lys Ser Gly    1650 1655 1660 Arg Leu Ala Leu Ala His Lys Thr Leu Val Leu Leu Leu Gly Val Asp 1665 1670 1675 1680 Pro Ser Arg Gln Leu Asp His Pro Leu Pro Thr Ala His Pro Gln Val                1685 1690 1695 Thr Tyr Ala Tyr Met Lys Asn Met Trp Lys Ser Ala Arg Lys Ile Asp            1700 1705 1710 Ala Phe Gln His Met Gln His Phe Val Gln Thr Met Gln Gln Gln Ala        1715 1720 1725 Gln His Ala Ile Ala Thr Glu Asp Gln Gln His Lys Gln Glu Leu His    1730 1735 1740 Lys Leu Met Ala Arg Cys Phe Leu Lys Leu Gly Glu Trp Gln Leu Asn 1745 1750 1755 1760 Leu Gln Gly Ile Asn Glu Ser Thr Ile Pro Lys Val Leu Gln Tyr Tyr                1765 1770 1775 Ser Ala Ala Thr Glu His Asp Arg Ser Trp Tyr Lys Ala Trp His Ala            1780 1785 1790 Trp Ala Val Met Asn Phe Glu Ala Val Leu His Tyr Lys His Gln Asn        1795 1800 1805 Gln Ala Arg Asp Glu Lys Lys Lys Leu Arg His Ala Ser Gly Ala Asn    1810 1815 1820 Ile Thr Asn Ala Thr Thr Ala 1825 1830 1835 1840 Ala Thr Ser Thr Glu Gly Ser Asn Ser Glu Ser Glu Ala Glu Ser Asn                1845 1850 1855 Glu Asn Ser Pro Thr Pro Ser Pro Leu Gln Lys Lys Val Thr Glu Asp            1860 1865 1870 Leu Ser Lys Thr Leu Leu Leu Tyr Thr Val Ala Val Gln Gly Phe        1875 1880 1885 Phe Arg Ser Ser Leu Ser Arg Gly Asn Asn Leu Gln Asp Thr Leu    1890 1895 1900 Arg Val Leu Thr Leu Trp Phe Asp Tyr Gly His Trp Pro Asp Val Asn 1905 1910 1915 1920 Glu Ala Leu Val Glu Gly Val Lys Ala Ile Gln Ile Asp Thr Trp Leu                1925 1930 1935 Gln Val Ile Pro Gln Leu Ile Ala Arg Ile Asp Thr Pro Arg Pro Leu            1940 1945 1950 Val Gly Arg Leu Ile His Gln Leu Leu Thr Asp Ile Gly Arg Tyr His        1955 1960 1965 Pro Gln Ala Leu Ile Tyr Pro Leu Thr Val Ala Ser Lys Ser Thr Thr    1970 1975 1980 Thr Ala Arg His Asn Ala Ala Asn Lys Ile Leu Lys Asn Met Cys Glu 1985 1990 1995 2000 His Ser Asn Thr Leu Val Gln Gln Ala Met Met Val Ser Glu Glu Leu                2005 2010 2015 Ile Arg Val Ala Ile Leu Trp His Glu Met Trp His Glu Gly Leu Glu            2020 2025 2030 Glu Ala Ser Arg Leu Tyr Phe Gly Glu Arg Asn Val Lys Gly Met Phe        2035 2040 2045 Glu Val Leu Glu Pro Leu His Ala Met Met Glu Arg Gly Pro Gln Thr    2050 2055 2060 Leu Lys Glu Thr Ser Phe Asn Gln Ala Tyr Gly Arg Asp Leu Met Glu 2065 2070 2075 2080 Ala Gln Glu Trp Cys Arg Lys Tyr Met Lys Ser Gly Asn Val Lys Asp                2085 2090 2095 Leu Thr Gln Ala Trp Asp Leu Tyr Tyr His Val Phe Arg Arg Ile Ser            2100 2105 2110 Lys Gln Leu Pro Gln Leu Thr Ser Leu Glu Leu Gln Tyr Val Ser Pro        2115 2120 2125 Lys Leu Leu Met Cys Arg Asp Leu Glu Leu Ala Val Pro Gly Thr Tyr    2130 2135 2140 Asp Pro Asn Gln Pro Ile Ile Arg Ile Gln Ser Ile Ala Pro Ser Leu 2145 2150 2155 2160 Gln Val Ile Thr Ser Lys Gln Arg Pro Arg Lys Leu Thr Leu Met Gly                2165 2170 2175 Ser Asn Gly His Glu Phe Val Phe Leu Leu Lys Gly His Glu Asp Leu            2180 2185 2190 Arg Gln Asp Glu Arg Val Met Gln Leu Phe Gly Leu Val Asn Thr Leu        2195 2200 2205 Leu Ala Asn Asp Pro Thr Ser Leu Arg Lys Asn Leu Ser Ile Gln Arg    2210 2215 2220 Tyr Ala Val Ile Pro Leu Ser Thr Asn Ser Gly Leu Ile Gly Trp Val 2225 2230 2235 2240 Pro His Cys Asp Thr Leu His Ala Leu Ile Arg Asp Tyr Arg Glu Lys                2245 2250 2255 Lys Lys Ile Leu Leu Asn Ile Glu His Arg Ile Met Leu Arg Met Ala            2260 2265 2270 Pro Asp Tyr Asp His Leu Thr Leu Met Gln Lys Val Glu Val Phe Glu        2275 2280 2285 His Ala Val Asn Asn Thr Ala Gly Asp Asp Leu Ala Lys Leu Leu Trp    2290 2295 2300 Leu Lys Ser Pro Ser Ser Glu Val Trp Phe Asp Arg Arg Thr Asn Tyr 2305 2310 2315 2320 Thr Arg Ser Leu Ala Val Met Ser Met Val Gly Tyr Ile Leu Gly Leu                2325 2330 2335 Gly Asp Arg His Pro Ser Asn Leu Met Leu Asp Arg Leu Ser Gly Lys            2340 2345 2350 Ile Leu His Ile Asp Phe Gly Asp Cys Phe Glu Val Ala Met Thr Arg        2355 2360 2365 Glu Lys Phe Pro Glu Lys Ile Pro Phe Arg Leu Thr Arg Met Leu Thr    2370 2375 2380 Asn Ala Met Glu Val Thr Gly Leu Asp Gly Asn Tyr Arg Thr Thr Cys 2385 2390 2395 2400 His Thr Val Met Glu Val Leu Arg Glu His Lys Asp Ser Val Met Ala                2405 2410 2415 Val Leu Glu Ala Phe Val Tyr Asp Pro Leu Leu Asn Trp Arg Leu Met            2420 2425 2430 Asp Thr Asn Thr Lys Gly Asn Lys Arg Ser Ser Thr Arg Thr Asp Ser        2435 2440 2445 Tyr Ser Ala Gly Gln Ser Val Glu Ile Leu Asp Gly Val Glu Leu Gly    2450 2455 2460 Glu Pro Ala His Lys Lys Ala Gly Thr Thr Val Pro Glu Ser Ile His 2465 2470 2475 2480 Ser Phe Ile Gly Asp Gly Leu Val Lys Pro Glu Ala Leu Asn Lys Lys                2485 2490 2495 Ala Ile Gln Ile Ile Asn Arg Val Arg Asp Lys Leu Thr Gly Arg Asp            2500 2505 2510 Phe Ser His Asp Asp Thr Leu Asp Val Pro Thr Gln Val Glu Leu Leu        2515 2520 2525 Ile Lys Gln Ala Thr Ser His Glu Asn Leu Cys Gln Cys Tyr Ile Gly    2530 2535 2540 Trp Cys Pro Phe Trp 2545 <210> 5 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> mTOR siRNA target <400> 5 caggcctatg gtcgagattt a   <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> rat ChREBP-α_Forward <400> 6 cgacactcac ccgcctcttc 20 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> rat ChREBP-α_Reverse <400> 7 ttgttcagcc gaatcttgtc 20 <210> 8 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> rat ChREBP-β_Forward <400> 8 tctgcagatc gcgcggag 18 <210> 9 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> rat ChREBP-β_Reverse <400> 9 cttgtcccgg catagcaac 19 <210> 10 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> rat TXNIP_Forward <400> 10 caagttcggc tttgagcttc 20 <210> 11 <211> 20 <212> DNA <213> Artificial Sequence <220> Rat TXNIP_Reverse <400> 11 acgatcgaga aaagccttca 20 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> mouse TXNIP_Forward <400> 12 tatgtacgcc cctgagttcc 20 <210> 13 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> mouse TXNIP_Reverse <400> 13 gctcactgca cgttgttgtt 20 <210> 14 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> rat TXNIP promoter_Forward <400> 14 cgcacccgaa caacaaccat 20 <210> 15 <211> 19 <212> DNA <213> Artificial Sequence <220> Rat TXNIP promoter_Reverse <400> 15 ggcaaaggcc gagggcgaa 19

Claims (5)

A pharmaceutical composition for the prevention or treatment of type 1 diabetes, comprising as an active ingredient an expression or activity inhibitor of Carbohydrate responsive element-binding protein (ChREBP)
Wherein the expression or activity inhibitor of ChREBP is a mTOR (mammalian Target Of Rapamycin) gene or an mTOR protein.
delete The method according to claim 1,
Wherein the mTOR gene comprises the nucleotide sequence of SEQ ID NO: 1.
The method according to claim 1,
Wherein the mTOR protein is comprised of the amino acid sequence of SEQ ID NO: 2.
The method according to claim 1,
Wherein the composition inhibits expression of TXNIP (Thioredoxin-interacting protein) by inhibiting expression or activity of ChREBP.

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