KR20220059990A - Composition for preventing or treating obesity or metabolic disease comprising XMU-MP-1 - Google Patents

Abstract

The present invention relates to a method for treating metabolic diseases through changes in the expression of STK3 or STK4 in fat cells, and in particular, relates to a composition containing XMU-MP-1, which promotes lipid metabolism to have effects of preventing, alleviating, and treating obesity or metabolic diseases, as an effective component. The composition of the present invention can be used for various purposes such as pharmaceutical and health functional food compositions.

Description

Composition for preventing or treating obesity or metabolic disease comprising XMU-MP-1

The present invention relates to a composition for preventing, improving or treating metabolic diseases such as obesity, diabetes, or hyperlipidemia comprising XMU-MP-1 as an active ingredient.

The Hippo signaling pathway is an intracellular signaling pathway that inhibits cell division and promotes cell death, thereby inhibiting the growth of body organs, and is known to be a pathway specifically involved in the regulation of adipogenesis. Various proteins and enzymes are involved in the Hippo signaling pathway, and they play an important role in determining the size of tissues and organs in animals. Regarding energy metabolism, dysregulation of the hippo kinase STK3/STK4 is strongly associated with the pathology of metabolic organs including liver, pancreas and skeletal muscle. In particular, in the beta cells of the pancreas, STK4 is upregulated in the conditions of diabetes mellitus, and is responsible for beta cell apoptosis. Based on these effects, STK3/STK4 inhibitors can be used as a preventive or therapeutic agent for diabetes.

It is also known that the change between brown and white fat increases energy expenditure and can be a target for treating obesity-related metabolic diseases. Among the Hippo signaling pathways, YAP/TAZ-dependent actomyosin responses mediate thermogenesis and browning in white fat, but do not require the activation of STK3/STK4, so their role in adipocytes is unclear.

Adipose tissue is a major metabolic organ that can be subdivided into two types: white adipose tissue (WAT) and brown adipose tissue (BAT). WAT is specialized for lipid storage and transport, while BAT is responsible for tremor-free thermogenesis. Although brown and white adipocytes are known to originate from distinct mesenchymal stem cell lineages, white adipocytes can be converted into brown-like adipocytes (so-called beige adipocytes) under thermogenic stimuli such as low temperature and beta-adrenergic stimulation. there is. This process, known as WAT browning, is characterized by an increase in oxidative mitochondrial metabolism that is closely related to mitophagy activity, along with the induction of uncoupling protein 1 (UCP1) expression and mitochondrial production.

STK3/STK4 is known to engage in various molecular-dependent interactions related to autophagy regulation in various tissues. Through this, it can be assumed that STK3/STK4 is related to energy metabolism in adipose tissue, and by regulating the mitochondrial activity of adipose tissue by regulating the expression of STK3/STK4, a therapeutic effect on obesity and metabolic diseases can be expected. It is expected that there will be

Accordingly, the present inventors demonstrated through this study that the adipocyte-specific deletion of STK3/STK4 increased the browning of white fat and prevented obesity. Through this, the present invention was completed on the basis that obesity and metabolic diseases can be treated by using a substance that regulates the expression of STK3/STK4.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing or treating obesity or metabolic disease, comprising XMU-MP-1 (Formula 1), which is a substance capable of regulating the expression of STK3/STK4.

Another object of the present invention is to provide a health functional food composition for preventing or improving obesity or metabolic disease, including XMU-MP-1 (Formula 1).

Another object of the present invention is to provide a method for screening a substance having an effect of preventing, improving and treating obesity or metabolic disease through the expression level analysis of STK3/STK4.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating obesity or metabolic disease comprising XMU-MP-1.

In order to achieve another object of the present invention, the present invention provides a health functional food composition for preventing or improving obesity or metabolic disease comprising XMU-MP-1.

In order to achieve another object of the present invention, the present invention provides a method for screening a substance having a preventive, ameliorating and therapeutic effect on obesity or metabolic disease through the expression level analysis of STK3/STK4.

Hereinafter, the present invention will be described in detail.

In the present invention, XMU-MP-1 is a substance represented by the structural formula of the following Chemical Formula 1, known as an ATP-competitive MST1/2 inhibitor, and is a substance known to have effects such as regeneration of liver tissue.

In the present invention, it was confirmed that XMU-MP-1 increased energy metabolism in adipose tissue and alleviated symptoms such as glucose intolerance by inhibiting the expression of STK3/STK4 specifically in adipocytes. It was confirmed that there is a preventive, ameliorating and therapeutic effect on obesity or metabolic disease. In one embodiment of the present invention, the expression level of STK3/STK4 was higher in white adipose tissue, which mainly stores fat, than in brown adipose tissue, which is responsible for energy metabolism such as heat generation, and when converted to brown fat by cold stimulation, STK3 It was confirmed that the expression of /STK4 was suppressed. In addition, it was confirmed that mitochondrial content was increased in adipose tissue due to the decrease in mitophagy expression in adipocyte-specific STK3/STK4 knockout (STK3/STK4 ^adipoq KO) mice. Consistent with the in vivo results, it was confirmed that genetic and pharmacological inhibition of STK3/STK4 in vitro reduced the turnover of mitochondria to mitophagy and increased mitochondrial content and respiration. In other words, the results show that STK3/STK4 ^adipoq KO increases energy expenditure and protects obesity and glucose intolerance caused by a high-fat diet.

In addition, in another embodiment of the present invention, it was confirmed that the up-regulation of STK3/STK4 in adipose tissue was observed in obese or type 2 diabetes patients. These results support the clinical relevance and pathological role of STK3/STK4 up-regulation with metabolic diseases such as obesity or diabetes.

In the present invention, the XMU-MP-1 inhibits adipocyte-specific expression of STK3/STK4, thereby having an effect of treating metabolic diseases such as obesity and diabetes. The metabolic disease is a generic term for diseases caused by imbalances such as carbohydrates, lipids, proteins, vitamins, minerals and water. , hypertension, hypercholesterolemia, and glucose intolerance. In particular, the composition comprising XMU-MP-1 of the present invention increases the conversion of fat to brown fat so as to increase thermogenesis and mitochondrial activity, and inhibits mitophagy activity. It has the effect of preventing, improving and treating various symptoms.

The pharmaceutical composition of the present specification includes all compositions capable of preventing or treating symptoms of metabolic diseases related to lipid metabolism.

As used herein, the term "prevention" refers to any action capable of suppressing or delaying the onset of symptoms related to metabolic diseases by administration of the composition according to the present invention.

As used herein, the term “treatment” refers to any action in which symptoms related to metabolic diseases are improved or beneficial by administration of the composition according to the present invention.

The pharmaceutical composition of the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, and sterile injection solutions according to conventional methods. and may further include a carrier or excipient necessary for the formulation. Pharmaceutically acceptable carriers, excipients and diluents that may be additionally included in the active ingredient include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate and mineral oil. In the case of formulation, it is prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

For example, solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include at least one or more excipients in the extract or compound at least cotton, starch, calcium carbonate , sucrose or lactose, gelatin, etc. are mixed and prepared. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, internal solutions, emulsions, syrups, etc. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. there is.

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like can be used.

The pharmaceutical composition of the present invention may be administered orally or parenterally (intravenous injection, subcutaneous, intraperitoneal or topical application) according to a desired method, and the dosage may vary depending on the patient's condition and weight, the degree of disease and the drug form; It depends on the route and time of administration, and may be selected in an appropriate form by those skilled in the art.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" is a reasonable amount applicable to medical treatment, means an amount sufficient to treat a disease, and the criteria are the patient's disease, severity, drug activity, sensitivity to drugs, administration time , administration route and excretion rate, treatment period, concomitant ingredients, and other factors. The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. Taking all of the above factors into consideration, the dosage can be determined at a level capable of minimizing side effects, which can be easily determined by those skilled in the art. Specifically, the dosage of the pharmaceutical composition may vary depending on the patient's age, weight, severity, sex, etc., and is generally 0.001 to 150 mg per 1 kg of body weight, more preferably 0.01 to 100 mg daily or every other day, 1 It may be administered 1 to 3 times a day. However, this is exemplary, and the dosage may be set differently if necessary.

In addition, the present invention relates to a health functional food composition for the prevention or improvement of obesity or metabolic disease, including XMU-MP-1, and the term "health functional food" in the present specification is defined in accordance with Health Functional Food Act No. 6727. It refers to food manufactured and processed using raw materials or ingredients with useful functionality to the human body. It means to consume for a purpose.

The food or health functional food of the present invention is a pharmaceutical dosage form such as powders, granules, tablets, capsules, pills, suspensions, emulsions, syrups, etc. or tea bags, leaches, beverages, for the purpose of preventing and improving metabolic diseases; It can be manufactured and processed into health functional foods such as candy, jelly, and gum.

The food or health functional food composition of the present invention may be used as a food additive, and may be commercialized alone or in combination with other ingredients. Also used in nutrients, vitamins, electrolytes, flavoring agents, colorants and enhancers, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonated beverages Carbonating agents and the like may be included. The above components may be used alone or in combination, and may be used in combination in an appropriate amount.

In addition, the present invention relates to a method for screening a substance having a preventive, ameliorating and therapeutic effect on obesity or metabolic disease through the expression level analysis of STK3/STK4, which comprises the steps of: i) treating adipose tissue with a candidate substance; ii) measuring the expression or activity of STK3 or STK4 in the adipose tissue treated with the candidate substance; and iii) selecting a substance that reduces the expression or activity of STK3 or STK4 as compared to the untreated group as having a preventive, ameliorating and therapeutic effect on obesity or metabolic disease.

In the present invention, the expression level analysis of STK3/STK4 can be performed through a variety of known protein analysis methods, for example, immunoprecipitation, immunostaining, chromatin immunostaining, enzyme immunoassay (ELISA), Western blotting, etc. It can be confirmed by a known method, but is not limited thereto.

The present invention relates to a composition comprising XMU-MP-1 as an active ingredient, which has an effect of preventing, improving and treating obesity or metabolic disease by promoting lipid metabolism through reduction in expression of STK3/STK4, the composition of the present invention can be used for various purposes such as pharmaceutical and health functional food composition.

1 shows the heat map analysis results of RNA-Seq (fetal brown fat (fBAT) and adult subcutaneous and mesenteric white fat (GEO accession number: GSE97205)) published from human adipose tissue.
2 is a comparison of the expression levels of STK4, STK3, HIF1A and PPARA in each adipose tissue (brown fat (fBAT), subcutaneous white fat (sub), mesenteric white fat (Omen)) among factors involved in the Hippo signal transduction pathway. one result is shown.
3 is a result showing the correlation between the expression levels of HIF1A and PPARA and STK3 and STK4.
4 shows the heat map analysis results of white fat (iWAT, GEO accession number: GSE13432) of the mouse groin induced by low temperature exposure.
5 shows the changes in the expression levels of STK4, STK3, HIF1A and PPARA in iWAT due to low temperature exposure.
6 shows the changes in the expression levels of STK4 and STK3 for each adipose tissue due to low temperature exposure.
7 is a qPCR analysis result of gWAT obtained from mice fed a high-fat diet (HFD) and a normal diet (NCD). (n = 10, mean ±SEM per group, *p<0.05, **p<0.01, ***p<0.001; t-test)
8 shows the results of indirect calorimetry analysis of wild-type and STK3/4 ^adipoq KO mice fed a high-fat diet.
9 shows the monitoring results of body fat mass and body weight of wild-type and STK3/4 ^adipoq KO mice fed a high-fat diet.
10 shows the results of intraperitoneal glucose tolerance testing of wild-type and STK3/4 ^adipoq KO mice fed a high-fat diet. (n=6 per condition, mean ± SEM, *p<0.05, **p<0.01, ***p<0.001)
11 is a result of measuring and comparing the amount of body fat between a mouse and a control group in which the expression of STK3/STK4 was suppressed by administering XMU-MP-1.
12 is a result of measuring and comparing the amount of change in body weight between a mouse and a control group in which the expression of STK3/STK4 was suppressed by administering XMU-MP-1.
13 shows the results of an intraperitoneal glucose tolerance test in mice and controls in which the expression of STK3/STK4 was suppressed by administering XMU-MP-1. (n=6 per condition, mean ± SEM, *p<0.05, **p<0.01, ***p<0.001)

Hereinafter, examples will be given to describe the present specification in detail. However, the embodiments according to the present specification may be modified in various other forms, and the scope of the present specification is not to be construed as being limited to the embodiments described below. The embodiments of the present specification are provided to more completely explain the present specification to those of ordinary skill in the art.

<Experiment method>

One. animal testing

All protocols related to animal experiments were performed in accordance with the regulations of Yonsei University (IACUC-A-201707-607-01) and Seoul National University (SNU-181205-2-1, SNU-200427-3, SNU-200720-1). . Mice were fed a normal diet (NCD) and maintained on a 12-hour light/dark cycle with free access to food and water at 22°C.

C57BL/6 mice (5-6 weeks of age) were obtained from Central Lab. Animal Inc. was purchased from Adipoq-CreER (B6.129-Tg(Adipoq-cre/Esrl*)1Evdr/J:stock #024671) mice and STK3/STK4 floxed mice (stock #017635) were purchased from Jackson Laboratory. mtKeima mice were obtained from Professor Jinho Yoon's laboratory at Dong-A University. Adipoq-CreER mice were crossed with STK3/STK4-floxed mice to generate adipocyte-specific STK3/STK4 KO mice. For Cre recombination, triple transgenic mice (Adipoq-CreER/STK3 ^fl/fl /STK4 ^fl/fl :STK3/STK4 KO) and double transgenic mice (STK3 ^fl/fl /STK4 ^fl/ fl without CreER: WT control group) ), tamoxifen (Sigma, 75 mg / kg) dissolved in sunflower oil was administered by oral gavage every 5 consecutive days. In addition, specific induction of Cre recombinase activity was confirmed upon tamoxifen treatment in adipose tissue, including vehicle (sunflower oil)-treated controls.

For the high fat diet (HFD) experiment, a 60% fat diet (D12492, Central Lab. Animal Inc.) was introduced at 7 weeks of age and continued for 8 weeks. For the cold tolerance test, the mice were stored in a refrigerator at 4°C after their body temperature was measured with a rectal thermometer as described in the previous work.

In addition, mtKeima mice were treated with XMU-MP-1 (1 mg/kg, i.p., Tocris, cat #6482, Bristol, UK) every 2 days for up to 10 days. Energy consumption was measured using an indirect calorimetry system (PhenoMaster, TSE system, Bad Homburg, Germany). Body composition was measured with an EchoMRI 700 (Echo Medical System) using nuclear magnetic resonance (NMR) scanning.

2. Quantitative PCR (qPCR) and Western blot analysis

Quantitative PCR and Western blot analysis were performed according to known methods. As the STK3/STK4 primer, a known primer was used.

3. cell culture

C3H10T1/2 (American Type Culture Collection (ATCC), Manassas, VA) cells were used for in vitro adipocyte experiments, and adipogenic differentiation was performed according to a known method. For siRNA knockdown (KD), fully differentiated C3H10T1/2 cells were transfected with 20 nM STK3/STK4 siRNA (Sigma, cat #58231-1 or custom design) using INTERFERin® (polyplus, cat #409-10). did

In addition, primary cultures of stromal cells and differentiated adipocytes obtained from STK3/STK4 ^adipoq KO mice were analyzed according to a known method. Fully differentiated adipocytes were cultured in 10% FBS DMEM containing 1 μM 4-hydroxytamoxifen (Sigma, H7904) for 3 days and maintained in 10% FBS DMEM without 4-hydroxytamoxifen for 2 days.

4. cell fraction

Freshly collected adipose tissue was homogenized and nuclear and cytoplasmic fractions were extracted using NE-PER nuclear and cytoplasmic extraction reagent (ThermoFisher Scientific, cat #78833).

5. Histology and Immunohistochemistry

Hematoxylin/eosin staining and immunostaining were performed using paraffin sections according to a known method.

6. statistical analysis

Statistical analysis was performed using GraphPad Prism 5 software (GraphPad Software, La Jolla, CA, USA). Data are expressed as mean±standard error of the mean (SEM), and statistical significance between the two groups was determined by unpaired t-test. Comparisons between multiple groups were performed using one-way or two-way analysis of variance (ANOVA) with Bonferroni post hoc tests to determine p-values. The heat map was generated by the well-known PermutMatrix program.

Experimental Example 1. Comparison of STK3 / STK4 expression levels in adipose tissue

1-1. Comparison of expression levels by adipose tissue

With the approval of the Chonbuk National University Hospital Institution Review Committee (Permission No.: CUH 2017-03-026), samples were collected from the patient's abdominal fat and analyzed.

Data of published RNA-Seq (fetal brown adipose (fBAT) and adult subcutaneous and mesenteric white fat (GEO accession number: GSE97205)) from human adipose tissue were compared, and genes related to the Hippo signaling pathway were analyzed through heat map analysis. was analyzed.

As a result, STK4 expression level was more abundant in adult mesenteric white adipocytes compared to fBAT, and STK3 expression level was more abundant in human mesenteric white fat than subcutaneous white fat. In addition, HIF1A and STK3/STK4, which have a low expression level in brown fat and high expression level in mesenteric white fat, generally have a positive correlation, and PPARA and STK3/STK4, which have a high expression level in brown fat, generally have a negative correlation. was confirmed (FIGS. 1 to 3).

In order to determine the association between browning of white fat and STK3/STK4, data from analysis of white fat (iWAT) in the groin of mice induced by exposure to low temperature (4°C) for 1 week were used (GEO Accession No.: GSE13432) . Upon low-temperature exposure, expression of STK3/STK4 was suppressed in white fat (iWAT) in the groin ( FIGS. 4 and 5 ). In addition, STK4 was more dominant in white fat than in brown fat of mice, and it was confirmed that the levels of STK3/STK4 were down-regulated due to low temperature exposure in all parts (FIG. 6).

That is, it was confirmed that the expression of STK3/STK4 was suppressed when white fat was induced into brown fat.

1-2. Analysis of expression level of STK3/STK4 according to the mouse diet

Mice were fed a regular diet and a high-fat diet, respectively, and mesenteric white adipose tissue (gWAT) was isolated from each, and the expression level of STK3/STK4 was analyzed through qPCR. As a result, as shown in FIG. 7 , it was confirmed that the expression level of STK3/STK4 was increased in mice fed a high-fat diet.

Experimental Example 2. Increase of energy consumption by adipocyte-specific STK3/STK4 knockout and protective effect of high-fat diet-induced obesity and glucose tolerance

2-1. increased energy consumption

Indirect calorimetry was performed using STK3/STK4 ^adipoq KO mice and wild-type mice to determine whether the increase in mitochondrial content in adipose tissue by STK3/STK4 knockout affects energy consumption. As a result of checking the consumption levels of VO ₂ , VCO ₂ and energy by feeding high-fat diet to wild-type mice and STK3/STK4 ^adipoq KO mice for 4 weeks, STK3/STK4 ^adipoq KO mice showed VO ₂ , VCO ₂ levels and energy compared to wild-type mice. It was confirmed that the consumption level increased, and the RER decreased. (Fig. 8)

2-2. Obesity suppression effect

For obesity induction in wild-type mice and STK3/STK4 ^adipoq KO mice, a high-fat diet and a regular diet were fed for 4 weeks, and weight gain and fat mass increase were confirmed.

As a result, body weight was increased in both STK3/STK4 ^adipoq KO and wild-type mice fed a high-fat diet, but the weight gain was lower in ^{STK3/STK4 adipoq KO} mice. It was confirmed that the body fat mass was less increased compared to the (Fig. 9)

2-3. Sugar intolerance protective effect

To confirm the effect on glucose intolerance induced by a high-fat diet, a glucose tolerance test (GTT) was performed. After the mice were fasted for 12 hours, fasting blood glucose was measured, and glucose (2.0 g/kg body weight, 20% (w/v) glucose solution) was intraperitoneally injected. Thereafter, blood glucose levels were measured at 0, 15, 30, 45, 60, 90, 120 and 150 minutes, respectively. Glucose tolerance was expressed by calculating the total area under each GTT curve (AUG).

As a result, it was confirmed that the STK3/STK4 ^adipoq KO mouse had a lower increase in blood glucose than the wild-type, and reduced glucose tolerance. (Fig. 10)

Experimental Example 3. When XMU-MP-1 administration suppressed adipocyte-specific STK3/STK4 expression, high-fat diet-induced obesity and glucose tolerance protective effect

3-1. Obesity suppression effect

An experiment was conducted to compare the effects of obesity induction in wild-type mice and mice in which the expression of STK3/STK4 was suppressed by administration of XMU-MP-1.

First, in order to suppress the expression of STK3/STK4 in mice that had been on a high-fat diet for 8 weeks, XMU-MP-1 (1mg/kg, i.p.) was administered every other day for 2 weeks, and weight gain and fat mass increase were confirmed.

As a result, in the case of the STK3/STK4 expression-inhibited mice, it was confirmed that the amount of body fat was reduced compared to the wild type (FIG. 11). In the case of body weight, it was increased along with the wild type for a certain period of time, but it was confirmed that it decreased around 10 days after feeding ( FIG. 12 ).

3-2. Sugar intolerance protective effect

To confirm the effect on glucose intolerance induced by a high-fat diet in wild-type mice and mice in which the expression of STK3/STK4 was suppressed by administration of XMU-MP-1, a glucose tolerance test (GTT) was performed. . The experiment was performed in the same manner as in Experimental Example 2-3, and the results are shown in FIG. 13 .

As shown in FIG. 13 , it was confirmed that the blood glucose level in the XMU-MP-1 administered mice was lower than that of the wild-type, and that the glucose tolerance (GTT) was also reduced compared to the wild-type.

So far, the present invention has been looked at with respect to preferred embodiments thereof. Those of ordinary skill in the art to which the present invention pertains will understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments are to be considered in an illustrative rather than a restrictive sense. The scope of the present invention is indicated in the claims rather than the foregoing description, and all differences within the scope equivalent thereto should be construed as being included in the present invention.

Claims (6)

A pharmaceutical composition for preventing or treating a metabolic disease comprising XMU-MP-1 of Formula 1 below.
[Formula 1]

According to claim 1,
The metabolic diseases include obesity, diabetes, hyperlipidemia, hypertension, hypercholesterolemia or glucose intolerance, a composition for preventing or treating metabolic diseases. According to claim 1,
The composition is a composition for preventing or treating metabolic diseases, characterized in that it promotes lipid metabolism in adipose tissue. According to claim 1,
The composition for preventing or treating metabolic diseases, characterized in that the composition inhibits the expression of STK3 or STK4 in adipocytes. A whole-strength functional food composition for the prevention or improvement of metabolic diseases, comprising XMU-MP-1 of Formula 1 below.
[Formula 1]

i) treating adipocytes to be analyzed with a candidate substance;
ii) measuring the expression or activity of STK3 or STK4 in adipocytes treated with the candidate substance; and
Compared to the untreated group, a method for preventing, improving, or screening a therapeutic substance for a metabolic disease, comprising selecting a substance that reduces the expression or activity of STK3 or STK4.

Images