JP2008001690A - Agonist of g protein coupled receptor, and medicine - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an agonist of G protein coupled receptor, and to provide several medicines containing the antagonist as an active ingredient. <P>SOLUTION: The agonist of G protein coupled receptor contains a thiazolidine-based compound represented by general formula (I) [wherein, R is a (substituted) lower alkyl] as an active ingredient. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to an agonist of a G protein-coupled receptor (GPCR) and various medicines using the same.

  GPR120 and GPR40 and related molecules are known as G protein-coupled receptors (GPCR) having fatty acid as a ligand.

  GPR41, GPR42 and the like are known as similar molecules of GPR40.

  GPR120 is known to exist in the intestinal tract, lungs, brain, and the like. When a cell having GPR120 present in the intestinal tract acts with a fatty acid as a ligand that activates GPR120, gutagon-like peptide 1 (GLP-1), cholecystokinin (CCK) and other intestinal hormone peptides may be released. Are known.

  These peptides control physiological functions related to feeding behavior, such as insulin secretion from pancreatic β cells, pancreatic juice secretion from the pancreas, bile secretion from the gallbladder, and appetite suppression to the center. ing. However, by administering a substance that activates the function of GPR120 to a living body, it is possible to prevent and treat diabetes by enhancing insulin secretion, treat digestive insufficiency by promoting digestive secretion, and prevent and treat obesity by suppressing appetite.

  By the way, thiazolidine compounds such as pioglitazone (pioglitazone) and troglitazone (troglitazone) bind to peroxisome proliferator-activated receptor gamma (PPARγ), which is a nuclear receptor type transcription factor of adipocytes, It is known to promote transcription of various molecules. As a result, it has been used to promote the differentiation of adipocytes, and as a result, improve insulin resistance and thus treat diabetes.

The main medicinal effects of thiazolidine compounds so far are (1) suppressing activation of adipocytes as a target (Non-patent Document 1), suppressing production of substances that induce insulin resistance, and improving insulin resistance ( Non-patent literature 2) Diabetes treatment by (2) Pancreatic β cells responsible for insulin secretion Similarly, β cells are removed from external factors that inhibit insulin secretion of β cells or induce cell death. Treatment of diabetes by maintaining β-cell function by protecting (Non-patent Document 3), (3) Enhancing liver lipid capture ability (Non-patent Document 4), hyperlipidemia treatment and accompanying treatment , Cardiovascular disease treatment, diabetes treatment, (4) suppression of liver stellate cell activation to suppress liver fibrosis (Non-Patent Document 5), liver fibrosis treatment and non-alcoholic steatohepatitis (non-alcoholic) steatohepatitis: NAS H) treatment, (5) activation of cells that cause inflammation such as macrophages and monocytes (Non-Patent Document 6) and suppression of cytokine production (Non-Patent Document 7) to treat inflammatory diseases and rheumatoid arthritis, (6) Cancer treatment by suppressing certain types of cancer growth (Non-patent Document 8) and the like.
J. Biol Chem. 272, 5637, 1907 Hauner H., Diabetes Metab. Res. Rev., 2002 Mar-Apr; 18 Suppl 2: S10-5) (Tan MH, Int. J. Clin. Pract. Suppl., 2000 Oct; (113): 54-62 Ishida H et al., Metabolism, 2004 Apr; 53 (4): 488-94. Sato O. et al., J. Biol. Chem., 2002 May 3; 277 (18): 15703-11 GuoYT et al., World J. Gastroenterol, 2005 Aug 14; 11 (30): 4735-9. Nature, 391, 79-82,1998 Nature, 391, 82-86,1998 Valentiner U et al., Toxicology, 2005 Sep 15; 213 (1-2): 157-68.

  An object of the present invention is to provide an agonist of a G protein-coupled receptor (GPCR) and a medicament containing the agonist as an active ingredient, such as a pancreatic beta differentiation regeneration promoter, a digestive tract disease therapeutic agent, a neuropathy therapeutic agent, The object is to provide a therapeutic agent for lung diseases, a therapeutic agent for metabolic syndrome, and the like.

  As a result of intensive studies to solve the above problems, the present inventor has obtained the following knowledge.

  Conventionally, thiazolidine compounds such as pioglitazone (pioglitazone) and troglitazone (troglitazone) bind to peroxisome proliferator-activated receptor gamma (PPARγ), which is a nuclear receptor type transcription factor of adipocytes, It is known to promote transcription of various molecules. As a result, it has been used to promote the differentiation of adipocytes, and as a result, improve insulin resistance and thus treat diabetes.

  The present inventors have found that such thiazolidine compounds can act (enhance) G protein-coupled receptors (GPCR) such as GPR120 (Sequence Tables 1 to 3) and GPR40 (Sequence Tables 4 and 5).

  As described above, GPR120 is present in the intestinal tract, lungs, brain, and the like, and cells possessing GPR120 present in the intestinal tract activate GPR120 with a thiazolidine-based compound so that glucagon-like peptide 1 (GLP-1), It was found that intestinal hormone peptides such as cystokinin (CCK) are released.

  Based on this knowledge, the present inventors have further studied and came to complete the present invention. That is, the present invention relates to an agonist of a G protein-coupled receptor (GPCR) containing a specific thiazolidine compound as an active ingredient, an appetite regulator containing the agonist, a pancreatic beta cell differentiation / regeneration / proliferation promoter, a digestive organ Provided are therapeutic agents for diseases, therapeutic agents for neurological disorders, and therapeutic agents for lung diseases. Specifically, the following aspects of the invention are provided.

（式中、Ｒは置換されていても良い低級アルキル基を示す。）
で表される化合物を有効成分として含有するものである。 The agonist of the G protein-coupled receptor of the present invention has the following general formula (I)

(In the formula, R represents an optionally substituted lower alkyl group.)
Is contained as an active ingredient.

The compound represented by the general formula (I) is preferably at least one selected from the group consisting of pioglitazone, troglitazone, ciglitazone, and rosiglitazone.

The G protein coupled receptor (GPCR) is preferably GPR120 or GPR40.

Drugs containing the above G protein-coupled receptor agonists as active ingredients include appetite regulators, pancreatic beta cell differentiation / regeneration / promoters, digestive diseases, neuropathies, psychiatric disorders, and lung diseases There are drugs and drugs for metabolic syndrome.

Hereinafter, the present invention will be described in detail.

  The G protein-coupled receptor (GPCR) agonist of the present invention contains a compound represented by the following general formula (I) (hereinafter referred to as a thiazolidine compound) as an active ingredient.

（式中、Ｒは置換されていても良い低級アルキル基を示す。）

(In the formula, R represents an optionally substituted lower alkyl group.)

  Examples of the lower alkyl group of the optionally substituted lower alkyl group represented by R include a linear or branched C1-6 alkyl group, preferably a methyl group, an ethyl group, an n-propyl group, and the like. is there.

  Examples of the substituent on the lower alkyl group in R include an optionally substituted pyridyl group, an optionally substituted amino group, an optionally substituted cycloalkyl group, and an optionally substituted chromanyl group. Can be mentioned.

  Examples of the substituent on the pyridyl group of the substituted pyridyl group include a C1-3 alkyl group, a C1-3 alkoxy group, and a hydroxyl group.

  Examples of the substituent on the amino group of the substituted amino group include a C1 to C3 alkyl group and a pyridyl group.

  Examples of the substituent on the cycloalkyl group of the substituted cycloalkyl group include C1-3 alkyl groups.

  Examples of the substituent on the substituted chromanyl group include a C1 to C3 alkyl group and a hydroxyl group.

  Preferable R is a group shown below.

  That is, preferable thiazolidine compounds include pioglitazone, troglitazone, ciglitazone, rosiglitazone, and the like.

  The G protein-coupled receptor (GPCR) agonist of the present invention contains the thiazolidine compound described above as an active ingredient. Conventionally, thiazolidine compounds bind to intracellular receptor PPARγ and promote transcription of various molecules, thereby promoting adipocyte differentiation, resulting in improvement of insulin resistance and thus treatment of diabetes. Has been used.

  However, this study revealed that some thiazolidine-based compounds act (enhance) GPR120 or GPR40 functions. Specifically, in the GPCR agonist of the present invention, a thiazolidine-based compound acts on intestinal hormone-secreting cells to release intestinal hormones, and these hormones (specifically, CCK and GLP-1) are received by target cells. It is based on a new mechanism that acts on the body (CCK receptor or GLP-1 receptor) and exhibits various medicinal effects. That is, the thiazolidine-based compound exerts a previously unknown medicinal effect based on a novel mechanism that is different from a conventional mechanism such as diabetes treatment. Moreover, since the mechanism is different and the target organ of the thiazolidine compound is also different, different medicinal effects are exhibited.

  The target disease and efficacy of the present invention are based on an increase in CCK concentration in the blood or target organ and an increase in GLP-1 concentration in the blood or target organ. In the present invention, GPCR is activated by a thiazolidine-based compound, and a wide range of treatments with an increase in the CCK or GLP-1 concentration in blood or in a target organ are possible.

  As a target disease associated with an increase in CCK concentration, for example, (i) promoting digestive activity. These include promotion of pancreatic juice secretion, promotion of gastric juice secretion, promotion of bile secretion, retention of food retention in the stomach, promotion of intestinal motility, and prevention of reflux due to contraction of the lower esophageal sphincter. Increases CCK and treats disorders of digestive activity. (ii) Gives a feeling of fullness and suppresses appetite. Increasing CCK suppresses appetite-related diseases such as obesity and treats bulimia nervosa. (iii) Promote differentiation and proliferation of cells in the gastric mucosa. Treatment of gastric wall disorders. (iv) Neural repair and treatment with nerve repair and maintenance. The description is not limited to this description, and the entire treatment with increased CCK concentration is targeted.

  On the other hand, as a target disease associated with an increase in the concentration of GLP-1, for example, (1) anti-diabetic drug for preventing transition from hyperglycemia, insulin resistance, obesity, etc. to diabetes by promoting differentiation and proliferation of pancreatic β cells . Or improved engraftment rate of transplanted cells at the time of β-cell transplantation; (2) Treatment with excessive gastric acid by inhibiting gastric acid secretion; (3) Treatment for diarrhea by inhibiting intestinal motility (4) Disease caused by neuropathy while maintaining nerve plasticity and survival And (5) obesity prevention treatment by suppressing appetite. The present invention is not limited to this description, and the entire treatment associated with an increase in the concentration of GLP-1 is targeted.

  Thiazolidine compounds release CCK and GLP-1 at the same time, so both of them often exhibit the same medicinal effect, but they appear to antagonize each other in digestive activity. In fact, both cooperate smoothly to perform digestive activity, and the simultaneous release of CCK and GLP-1 is a more physiologically rational treatment.

  The effect of thiazolidine compounds directly on the target organ is naturally different from the case where CCK and GLP-1 are released via GPR120 (Sequence Listings 1 to 3) and these act on the target organ. In particular, if the medicinal effects different from the conventional direct action are described, the application expansion is as follows. (I) cooperative promotion of digestive activity, treatment of digestive activity disorder, (ii) obesity prevention treatment by appetite suppression, bulimia treatment, (iii) diabetes prevention treatment by promoting pancreatic β cell differentiation and proliferation. In particular, β-cell or progenitor cell transplantation treatment effect accelerator, (iv) nerve cell plasticity, nerve transplantation by survival maintenance action, nerve junction treatment accelerator or Alzheimer's disease treatment caused by neuronal cell disorders , (V) treatment of intestinal motility abnormalities during enteritis due to normalization of intestinal motility, (vi) inhibition of intestinal cell damage or inhibition of nerve cells present in the intestinal tract, (vii) improvement of lung function in the lung, eg surfactant secretion Treatment of pulmonary diseases such as COPD (chronic obstructive pulmonary disease) by promotion, (viii) GPR120 agonists promote both CCK and GLP-1 actions simultaneously, so that obesity, insulin resistance, hyperglycemia, etc. are comprehensive Metabolic syndrome treatment, etc. by treatment.

  On the other hand, the thiazolidine-based compounds that actuate GPR40 (Sequence Listing 4, 5) acted on GPR40 as any of the measuring agents, but the strength of the effect varied depending on the substance (FIG. 1). GPR40 is present in pancreatic β cells, and insulin secretion is promoted by the action of GPR40. The GPR40 agonist thiazolidine compound acts directly on β cells simultaneously with the conventional action of actuating PPARγ to suppress adipocyte activation and promotes differentiation and proliferation of pancreatic β cells. From this viewpoint, as a new drug effect expansion of thiazolidine compounds, diabetes preventive drugs that prevent the transition from hyperglycemia, insulin resistance, obesity, etc. to diabetes by promoting differentiation and proliferation of pancreatic β cells, or transplanted cells at the time of β cell transplantation It is possible to improve the survival rate.

  The agonist of GPCR containing the thiazolidine compound of the present invention as an active ingredient has the above-mentioned medicinal effects, and can be formulated, for example, as follows.

  The GPCR agonists of the invention are formulated to be compatible with therapeutically relevant routes of administration, including intravenous and oral administration. Solutions or suspensions used for intravenous administration include, but are not limited to, sterile diluents such as water for injection, saline solution, non-volatile oil, polyethylene glycol, glycerin, propylene glycol, or Other synthetic solvents, preservatives such as benzyl alcohol or other methylparaben, antioxidants such as ascorbic acid or sodium bisulfite, soothing agents such as benzalkonium chloride and procaine hydrochloride, chelates such as ethylenediaminetetraacetic acid (EDTA) Agents, buffers such as acetate, citrate, or phosphate, and agents for osmotic pressure adjustment such as sodium chloride or dextrose may be included.

  The pH can be adjusted with acids or bases such as hydrochloric acid or sodium hydroxide. Non-caliber preparations are contained in ampoules, glass or plastic disposable syringes or multidose vials.

A formulation suitable for injection is a sterile injectable solution or dispersion medium, a sterile aqueous solution (water-soluble) or dispersion medium and a sterile powder (lyophilized protein) that are prepared at the time of use. , Nucleic acids and the like). For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, CREMOPHOR EL ^™ (BASF, Parsippany, NJ), or phosphate buffered saline (PBS). When used as an injection, the GPCR agonist must be sterilized and retain sufficient fluidity to be administered using a syringe. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures. For example, using a coating agent such as lectin, maintaining a required particle size in the dispersion medium, and maintaining a proper fluidity by using a surfactant. Various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, ascorbic acid, and thimerosal can be used to prevent microbial contamination. In addition, polyalcohols such as sugar, mannitol, sorbitol, and agents that maintain isotonicity such as sodium chloride may be included in the composition. Compositions that can delay adsorption include agents such as aluminum monostearate and gelatin.

  Sterile injectable solutions are prepared by adding the required ingredients alone or in combination with other ingredients, then adding the required amount of active compound in a suitable solvent and sterilizing. Generally, a dispersion medium is prepared by incorporating the active compound into a sterile medium that contains a basic dispersion medium and the other necessary ingredients described above. Sterile powder preparation methods for the preparation of sterile injectable solutions include vacuum drying and lyophilization to prepare a powder containing the active ingredient and any desired ingredients derived from the sterile solution. It is.

  In the case of an oral preparation, an inert diluent or a carrier that does not cause harm even when taken into the body is included. Oral formulations are, for example, contained in gelatin capsules or compressed into tablets. For oral treatment, the active compound is incorporated with excipients and used in the form of tablets, troches, or capsules. Oral preparations can also be prepared using a flowable carrier. In addition, pharmaceutically compatible binding agents, and / or adjuvant materials may be included.

  Tablets, pills, capsules, lozenges and the like can contain any of the following ingredients or compounds with similar properties. Excipients such as microcrystalline cellulose, binders such as gum arabic, tragacanth or gelatin; swelling agents such as starch or lactose, alginic acid, PRIMOGEL or corn starch; lubricants such as magnesium stearate or STRROTES; colloidal silicon Lubricants such as dioxides; sweeteners such as sucrose or saccharin; or flavoring agents such as peppermint, methyl salicylic acid or orange flavor.

  When a preparation for systemic administration is used, it can be transmucosally or transdermally. For transmucosal or transdermal administration, penetrants that can penetrate the target barrier are selected. Transmucosal penetrants include surfactants, bile salts, and fusidic acid derivatives. Nasal sprays or suppositories can be used for transmucosal administration. For transmucosal administration, the active compounds are formulated in ointments, ointments, gels or creams.

  Thiazolidine-based compounds can also be prepared in the form of suppositories (eg, with bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.

  In the case of a controlled release preparation, it can be prepared using a carrier that can prevent immediate removal from the body. For example, biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used. Such materials are available from ALZA Corporation (Mountain View, CA) and NOVA Pharmaceuticals, Inc. (Lake Elsinore, CA) and can also be readily prepared by one of ordinary skill in the art. Liposomal suspensions can also be used as pharmaceutically acceptable carriers. Useful liposomes are prepared as a lipid composition comprising, but not limited to, phosphatidylcholine, cholesterol and PEG-derivatized phosphatidylethanol (PEG-PE) through a filter of appropriate pore size so as to be of a size suitable for use. Purified by evaporation. For example, Fab 'fragments of antibodies and the like may be bound to liposomes via a disulfide exchange reaction (Martin and Papahadjopoulos, 1982). See, for example, the description in Epstein et al., 1985; Hwang et al., 1980 for detailed preparation methods.

  The dose of the GPCR agonist of the present invention depends on the condition of the patient (human) or animal to be administered, the administration method, etc. in the treatment or prevention of a specific disease, but can be easily optimized by those skilled in the art. It is possible.

  For example, in the case of injection administration, for example, it is preferable to administer about 0.1 μg / kg to 500 mg / kg of the patient's body weight per day, and it will generally be possible to administer in one or more divided doses. Preferably, the dosage level is about 0.1 μg / kg to about 250 mg / kg per day, more preferably about 0.5 to about 100 mg / kg per day.

  For oral administration, it is preferably provided in the form of a tablet containing 1.0 to 1000 mg of active ingredient. Preferably, the dosage of the active ingredient for the patient (human) or animal to be treated is 0.01-100 mg / kg. The compounds are administered on a regimen of 1 to 4 times daily, preferably once or twice daily.

  Further, as a means for screening a thiazolidine compound, by measuring the operability to GPR120 and GPR40, it is possible to screen a better drug for the target diseases whose application is expanded as described here.

  The agonist of the G protein-coupled receptor (GPCR) of the present invention is effective as an appetite regulator, a pancreatic beta-differentiated cell proliferation-promoting agent, a gastrointestinal disease therapeutic agent, a neuropathy therapeutic agent, and a lung disease therapeutic agent.

  Hereinafter, the present invention will be described in more detail based on examples, but the present invention is not limited to these examples.

-Example 1 (Ligand Screening of GPR40 and GPR120 by extracellular signal-regulated kinase (ERK) Assay)-
The materials and methods used in this example are as follows.
1. Protein recovery (1) In this experiment, cell lines TXGPR40 and TXGPR120 were used, and seeded in a 35 mm dish at a cell number of 5 × 10 ⁵ / dish. After confirming colonization with Dish, Doxycyclin treatment (10 μg Doxycyclin was added to the cell solution) was performed, and Starvation (replaced with FBS (-) Medium, with Dox) was performed 28 hours before, and then assayed 20 hours later.
In addition, TXGR40, TXGR120, respectively, a cell line that stably expresses GPR40 or GPR120 protein by introducing a gene into which a human GPR40 (hGPT40) or human GPR120 (hGPR120) gene is incorporated into an expression vector that is expressed by Doxycycline treatment. means.
(2) Four types of Ligands were used: pioglitazone, troglitazone, ciglitazone, and rosiglitazone. These were prepared by dissolving in DMSO and diluted 10-fold with Medium to make a total volume of 100 μl. Further, DMSO was used as a negative control, and PMA (Phorbol-12-myristate-13-acetate) was used as a positive control.
(3) Medium was removed from the dish of cells, and the total liquid volume was adjusted to 0.9 ml.
(4) Ligand solution was dripped from above the cell solution, and the dish was gently shaken to diffuse and allowed to react at room temperature.
(5) After the reaction for 5 minutes, the medium was discarded, and the medium remaining in the dish was removed on ice with an aspirator.
(6) 150 μl Lysis buffer (50 mM HEPES (pH 7.0), 150 mM NaCl, 10% glycerol, 1% NonidetP-40, 2 mM MgCl ₂ , 1 mM EDTA, 100 mM NaF, 10 mM Sodium phosphate, 1 mM Na ₃ VO ₄ , 20 mM β- glycerophosphate, Proteinase Inhibitor) was added, and the cells were recovered with Cell Scraper.
(7) The recovered solution is mixed with 150 μl (equal volume) of 1 × SDS Sample Buffer (0.1M Tris-HCl (pH 6.8), 4% SDS, 12% Mercaptoethanol, 20% glycerol, BPB) Used for Western Blotting or stored at -20 ° C.

2. SDS-PAGE
(1) The crude enzyme solution mixed with SDS Sample Buffer was sonicated using sonifier and then incubated at 90 ° C. for 5 min.
(2) Vortexed and centrifuged (15,000 rpm, 5 min, 4 ° C.).
(3) The electrophoresis tank was filled with 1 × SDS-PAGE Running Buffer (25 mM Tris, 0.2 M Glycine, 1% SDS), two 7.5% acrylamide gels were set, and the supernatant after sample centrifugation was applied. Electrophoresis was performed at a constant current of 40 mA. (About 80 ~ 90min)

3. Blotting (Semi-Dry method)
(1) PVDF membrane is cut according to the size of the gel, soaked in methanol for about 15 seconds, and then shaken in Transfer Buffer C solution (25 mM Tris, 0.02% SDS, 2% methanol, 40 mM 6-aminohexanoic acid) for 15 min ( Room temperature) to swell. The filter paper used for blotting was cut into a size of 10 cm × 10 cm.
(2) Two filter papers soaked in A solution (0.3M Tris, 0.02% SDS, 2% methanol) and two filter papers soaked in B solution (25mM Tris, 0.02% SDS, 2% methanol) I put a membrane on it. Further, the gel after the electrophoresis was placed thereon, and finally two filter papers soaked in C solution were stacked and covered.
(3) Transfer was performed at 15 V for 30 minutes.

4). Blocking
(1) All the following operations are at room temperature. After completion of the transfer, the membrane was transferred to a case containing 1 × TTBS (0.2 M Tris, 1.5 M NaCl, 0.2 & Tween 20) and washed for 15 minutes.
(2) Excluding TTBS, about 10 ml of Block Ace was added and shaken for 1 hour to block.

5. Primary antibody (1) Primary antibodies (p44 / 42 MAP Kinase Antibody and Phospho-p44 / 42 MAP Kinase Antibody) were each diluted 1000 times with TTBS.
(2) After the completion of Blocking, the Block Ace solution was removed, the primary antibody solution was added, and the reaction was performed by shaking for 2 hours.

6). Secondary antibody (1) The secondary antibody (Anti-rabbit Ig, Horseradish Peroxidase linked nF (ab ') ₂ fragment) was diluted 5000 times with TTBS.
(2) The primary antibody solution was removed and washed in TTBS by shaking for 5 to 10 minutes. Wash 3 times.
(3) TTBS was removed and the secondary antibody solution was added and shaken for 1 hour.

7). Detection (1) 2 ml each of Detection Reagent 1 and Detection Reagent 2 in ECL kit were mixed on ice.
(2) The secondary antibody solution was removed from the membrane after completion of the secondary antibody reaction, and the membrane was shaken and washed in TTBS for 5 minutes. Wash 3 times.
(3) The membrane was thoroughly drained into the pack, and 2 ml of Detection Reagent mixed solution was applied to each membrane.
(4) The membrane and film were brought into contact with each other for 1 to 5 minutes in a cassette and developed with an automatic processor.

8). Analysis (1) The film was scanned and loaded into a personal computer, and the band was digitized using the “Image J” application (Image J 1.345: free software published by National Institutes of Health).
(2) Normalize with DMSO and PMA and compare values between drugs.

  TXGPR40 and TXGPR120 were reacted with pioglitazone, troglitazone, siglitazone and rosiglitazone at concentrations of 10 μM and 100 μM. The signal was digitized and normalized by DMSO and PMA.

  The results of TXGPR40 and TXGPR120 are shown in FIGS. 1 and 2, respectively. In the data of each substance shown in FIG. 1 and FIG. 2, A (Doxt) indicates the reaction of cells expressing GPR protein by Doxycycline treatment, and B (Dox-) expresses GPR protein without Doxycycline treatment. C (TXCONT) indicates the response of a cell into which only a vector containing no GPR gene has been introduced. The following knowledge was obtained from the data of FIGS.

  TXCONT without GPR integration and cells not treated with Doxycycline (Dox (-)) were arranged as negative controls, and ERK activity against 4 types of Ligand was compared for those treated with Doxycycline (Dox (+)). . As for hGPR40, a reaction was observed with any Ligand. On the other hand, hGPR120 reacted only with troglitazone and ciglitazone, and other Ligands hardly activated ERK.

  Thereby, it can be understood that pioglitazone, troglitazone, ciglitazone and rosiglitazone are specifically reacting with cells in which GPR40 (sequence tables 4 and 5) is induced and expressed. Therefore, it is suggested that the above four compounds are suitably used as GPR40 agonists.

  Moreover, it can be understood that troglitazone and siglitazone are specifically reacting with cells in which GPR120 (sequence tables 1 to 3) is induced and expressed. Therefore, it is suggested that the above two compounds are suitably used as GPR120 agonists (agonists).

-Example 2-
C57BL / 6 CrSlc (8 weeks old, sputum, approximately 25 g) was administered with a Nembutal solution (the stock solution was diluted to 1/10 with physiological saline) and anesthetized. After laparotomy, the administered substance was suspended in 100 μl of 100 nmol / g body weight polyethylene glycol (PEG: average molecular weight 400, SIGMA) and administered to the intestine (colon). The mixture was allowed to stand for 15 minutes after administration, and blood was collected from the portal vein. The collected blood sample was centrifuged, and the plasma was stored frozen. Plasma GLP-1 concentration was measured using ELISA (rat GLP-1 ELISA kit Wako: Wako Pure Chemical Industries, Ltd.). Five mice were used for each group. α-Linolenic acid (SIGMA) and troglitazone (SIGMA) were used.

  FIG. 3 shows the average concentration (ng / ml) of GLP-1 when PEG alone, α-linolenic acid (α-LA) PEG suspension, and troglitazone PEG suspension are used as administration substances in Example 2, respectively. It is a figure shown as a table. From this result, it can be seen that troglitazone releases GLP-1 at the same level as α-LA, which is a GPR120 ligand, from the intestinal tract and is effective in activating GPR120 in cells.

GPR120 agonists or antagonists promote the release of GLP-1 or CCK from GPR120-bearing cells as shown in papers and patents (Nature Medicine, 11 (1), 90-94, 2005 and JP 2005-15358). As shown
(I) Cooperative promotion of digestive activity, therapeutic agent for digestive activity disorder. Nutrition 2001; 17 (3): 230-5, Best Pract Res Clin Endocrinol Metab 2004; 18 (4): 569-86, Dig Dis Sci. 2004; 49 (3): 361-9, Endocrinology 2004; 145 (6 ): 2653-9, Pharmacol Toxicol 2002; 91 (6): 375-81, Gastroenterology 2004; 127 (3): 957-69, Best Pract Res Clin Endocrinol Metab 2004; 18 (4): 569-86, Med Res Rev. 2003; 23 (5): 559-605, Med Res Rev.2003; 23 (5): 559-605, Horm Metab Res 2004; 36 (11-12): 842-5, Horm Metab Res 2004; 36 (11-12): 842-5, Am J Phyusiol Endcrinol Metab 2004; 287 (6): E1209-15, Dig Dis Sci 1998; 43 (4): 799-805, (II) Anti-obesity drug for suppressing appetite , Bulimia remedy. Physiol Behav. 2004; 83 (4): 617-21, Best Pract Res Cli Endocrinol Metab. 2004; 18 (4): 569-86, Trends Endcrinol Metab. 2004; 15 (6): 259-63, Cuur Drug Target CNS Neurol disord 2004; 3 (5): 379-88
(III) A prophylactic and therapeutic agent for diabetes by promoting differentiation and proliferation of pancreatic β cells. Therapeutic effect promoter especially at the time of β cell or its precursor cell transplantation treatment Diobetology, 2005; 48 (9): 1700-13, Diabetes 2004; 53suppl 3: S225-32, Hormone Metab Res. 2004 (11-12): 766 -70, Hormone Metab Res. 2004, 36 (11-12): 846-51
(IV) Nerve cell plasticity, nerve transplantation by viability maintenance action, treatment accelerator at the time of nerve junction, or therapeutic agent for diseases caused by nerve cell disorders such as Alzheimer's disease. Curr Drug Target CNS Neurol Disord 2002; 1 (5): 495-510, Curr Drug Targets. 2004; 5 (69: 565-71, Curr Alzheimer Res 2005, 2 (3): 377-85
(V) Drug for treatment of intestinal motility during enteritis due to normalization of intestinal motility Drug 2003; 63 (12): 1785-97, Br J Pharmacol. 2004; 141 (8): 1275-84,
(VI) Endocrinology. 1998; 139 (5): 2363-8. Am J Respir Crit Care Med. 2001; 163 (4): COPD (chronic obstructive pulmonary disease) by promoting the secretion of surfactant in the lung 840-6.
(VII) Metabolic syndrome drug. The above-mentioned documents of Nature Medicine, 11 (1), 90-94, 2005 and JP-A-2005-15358 and (II) are combined.

2 is a graph showing the results of GPR40 ligand screening by ERK assay in Example 1. FIG. 2 is a graph showing the results of ligand screening of GPR120 by ERK assay in Example 1. FIG. It is a figure which shows the GLP-1 average density | concentration with respect to various administration substances in Example 2 in a table | surface.

Claims (11)

Formula (I):

(In the formula, R represents an optionally substituted lower alkyl group.)
A G protein-coupled receptor (GPCR) agonist comprising a compound represented by the formula:   The compound represented by the general formula (I) is at least one selected from the group consisting of pioglitazone, troglitazone, ciglitazone, and rosiglitazone. G protein-coupled receptor agonist.   The G protein-coupled receptor agonist according to claim 1, wherein the G protein-coupled receptor (GPCR) is GPR120.   The G protein-coupled receptor agonist according to claim 1, wherein the G protein-coupled receptor (GPCR) is GPR40.   An appetite regulator comprising the G protein-coupled receptor agonist according to any one of claims 1 to 4 as an active ingredient.   A pancreatic beta cell differentiation / regeneration / proliferation promoter containing the G protein-coupled receptor agonist according to any one of claims 1 to 4 as an active ingredient.   A therapeutic agent for digestive system diseases comprising the G protein-coupled receptor agonist according to any one of claims 1 to 4 as an active ingredient.   A therapeutic agent for neuropathy comprising the G protein-coupled receptor agonist according to any one of claims 1 to 4 as an active ingredient.   A therapeutic agent for mental disorders containing the G protein-coupled receptor agonist according to any one of claims 1 to 4 as an active ingredient.   A therapeutic agent for lung diseases comprising the G protein-coupled receptor agonist according to any one of claims 1 to 4 as an active ingredient.   A therapeutic agent for metabolic syndrome comprising the G protein-coupled receptor agonist according to any one of claims 1 to 4 as an active ingredient.

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