Novel compound having Anti-obesity action

Abstract

To provide a new compound useful for promoting the decomposition of neutral fat in fat cells, and to provide a new therapeutic agent for obesity and a new therapeutic agent for obesity-related diseases, each containing the compound as an active ingredient.SOLUTION: There are provided a compound represented by formula (I) or a pharmaceutically acceptable salt or solvate thereof, and a therapeutic agent for obesity and a therapeutic agent for obesity-related diseases comprising the same as an active ingredient. (wherein, X and Z represent N or C, R1 represents alkyl or the like, R2 and R3 represent hydrogen, alkoxycarbonyl, alkylcarbamoyl or the like, R4 and R7 represent hydrogen, R5 and R6 represent hydrogen, alkoxycarbonyl, alkylcarbamoyl or the like) SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a novel compound having anti-obesity activity and a pharmaceutical composition comprising the same. [Background technology]

[0002] Obesity is a condition characterized by the excessive accumulation of fat in the body. It is characterized by hyperglycemia, hyperlipidemia, and hypertension. The Japan Society for the Study of Obesity defines "obesity" as a body mass index (BMI) of 25 or higher. Obesity in adults is caused by overeating and decreased activity, resulting in hypertrophy of fat cells at the cellular level. Compared to non-obese individuals, obese individuals are generally more susceptible to a variety of diseases, often with dysglycemia and dyslipidemia, which increases the risk of conditions such as dysglycemia, cardiovascular disorders, hypertension, sleep apnea syndrome, and osteoarthritis. Obesity treatments include diet and exercise therapy, surgical procedures such as jejunoileostomy, and pharmacotherapy with anti-obesity drugs.

[0003] To date, lipase inhibitors, appetite suppressants, and β-adrenergic antagonists have been developed as anti-obesity drugs, which have the effect of suppressing inappropriate weight gain and visceral fat gain (Patent Documents 1 and 2). Furthermore, cAMP and its derivatives are known as compounds that promote lipolysis (Non-Patent Documents 1 to 5), but no other promising compounds are known. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] International Publication No. 2006 / 129785 [Patent Document 2] Japanese Patent Application Laid-Open No. 2000-80047 [Non-patent literature]

[0005] [Non-Patent Document 1] Journal of Molecular Endocrinology, 2014, 52: R199-R222 [Non-patent document 2] Japanese Journal of Pharmacology, 2015, 146: 93-97 [Non-patent document 3] Nature Metabolism, 2021, 3: 1445-1465 [Non-patent document 4] Gene, 2011, 477: 1-11 [Non-Patent Document 5] Showa University Journal of Pharmaceutical Sciences, 2010, 1: 39-51 Summary of the Invention [Problem to be solved by the invention]

[0006] An object of the present invention is to provide a novel compound useful for promoting the decomposition of neutral fat in adipocytes, and a novel therapeutic agent for treating obesity and obesity-related diseases containing the compound as an active ingredient. [Means for solving the problem]

[0007] The present inventors have now found that the compound of formula (I) sustainably promotes the breakdown of neutral fat in adipocytes. The present inventors have also found that, when used in combination with a cAMP analog, the compound of formula (I) synergistically enhances the neutral fat breakdown-promoting effect of the cAMP analog. The present inventors have also found that the compound of formula (I) activates protein kinase A and hormone-sensitive lipase. The present inventors have also found that administration of the compound of formula (I) to mice sustainably promotes the breakdown of neutral fat and reduces body weight. The present invention is based on these findings.

[0008] According to the present invention, the following inventions are provided. [1] A compound of the following formula (I) or a pharmaceutically acceptable salt or solvate thereof (hereinafter, sometimes referred to as "the compound of the present invention"). [ka] (In the above formula, X and Z represent carbon atoms or nitrogen atoms, and when X represents a nitrogen atom, R 5 does not exist, and when Z represents a nitrogen atom, R 6 does not exist, and both X and Z do not represent nitrogen atoms. --- represents a single or double bond, R 1 is a hydrogen atom, an alkyl group having 1 to 4 carbon atoms, an alkylcarbonyl group having 1 to 4 carbon atoms, or -R a -R b (In the formula, R a represents -(CH2)m- (m is an integer of 1 to 3), one -(CH2)- in -(CH2)m- may represent -(C=O)-, R b represents an aromatic ring group or an aliphatic ring group, and the aromatic ring group and the aliphatic ring group may be substituted with an alkoxy group having 1 to 4 carbon atoms, an alkyl group having 1 to 4 carbon atoms, or a halogen atom; R 2 and R 3 may be the same or different and are a hydrogen atom or -(C=O)-R c (In the formula, R c represents a hydroxyl group, an alkoxy group having 1 to 4 carbon atoms, -N(-R 11 )(-R 12 )(wherein, R 11 and R 12 each represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms) or -O-(CH2)n-(C=O)-R 13 (wherein n is an integer of 1 to 3, and R 13 represents an alkyl group having 1 to 4 carbon atoms), where R 2 and R 3 does not simultaneously represent a hydrogen atom, R 4 and R 7 represents a hydrogen atom, R 5 is a hydrogen atom or -(C=O)-R d (In the formula, R d represents a hydroxyl group, an alkoxy group having 1 to 4 carbon atoms, -N(-R21 )(-R 22 )(wherein, R 21 and R 22 each represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms) or -O-(CH2)p-(C=O)-R 23 (wherein p is an integer of 1 to 3, and R 23 represents an alkyl group having 1 to 4 carbon atoms), R 6 is a hydrogen atom or -(C=O)-R e (In the formula, R e represents a hydroxyl group, an alkoxy group having 1 to 4 carbon atoms, -N(-R 31 )(-R 32 )(wherein, R 31 and R 32 each represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms) or -O-(CH2)r-(C=O)-R 33 (wherein r is an integer of 1 to 3, and R 33 represents an alkyl group having 1 to 4 carbon atoms. [2]R 1 is an alkyl group having 1 to 4 carbon atoms or -R a -R b (In the formula, R a represents -CH2- or -C(=O)-, and R b represents an optionally substituted aromatic ring group or an optionally substituted aliphatic carbocyclic group), or a pharmaceutically acceptable salt or solvate thereof. [3] --- represents a double bond, or a pharmaceutically acceptable salt or solvate thereof. [4]R 2 and R 3 may be the same or different and represent an alkoxycarbonyl group having 1 to 4 carbon atoms, or a pharmaceutically acceptable salt or solvate thereof according to any one of the above [1] to [3]. [5] X represents a nitrogen atom, Z represents a carbon atom, and R 5 does not exist and R 6represents an alkoxycarbonyl group having 1 to 4 carbon atoms, or a pharmaceutically acceptable salt or solvate thereof. [6] An agent for promoting neutral fat degradation in adipocytes, comprising, as an active ingredient, the compound of formula (I) described in any one of the above [1] to [5] or a pharmaceutically acceptable salt or solvate thereof. [7] A pharmaceutical composition comprising, as an active ingredient, the compound of formula (I) described in any one of [1] to [5] above, or a pharmaceutically acceptable salt or solvate thereof. [8] The pharmaceutical composition according to [7] above, for use in treating, preventing, or ameliorating a disease or symptom that can be treated, prevented, or ameliorated by promoting neutral fat breakdown in adipocytes. [9] The pharmaceutical composition according to [8] above, wherein the disease or symptom is obesity or a disease or symptom associated therewith.

[10] The pharmaceutical composition according to [9] above, wherein the obesity-related disease and symptom are one or more selected from the group consisting of impaired glucose tolerance, dyslipidemia, hypertension, hyperuricemia, gout, coronary artery disease, cerebral infarction, fatty liver, menstrual disorders, pregnancy complications, sleep apnea syndrome, obesity-hypoventilation syndrome, orthopedic diseases, and obesity-related kidney disease.

[11] The pharmaceutical composition according to any one of [7] to

[10] above, which is used in combination with an agent for promoting cAMP production in adipocytes.

[0009] According to the present invention, there are provided novel compounds useful for promoting the decomposition of neutral fats in adipocytes, and novel anti-obesity drugs and therapeutic drugs for obesity-related diseases that contain the compounds as active ingredients. [Brief explanation of the drawings]

[0010] [Figure 1] FIG. 1 is a graph showing the effect of the compound of the present invention on the production of free fatty acids from adipocytes (Test Example 2). [Figure 2] FIG. 2 is a graph showing the effect of the compound of the present invention on hormone-sensitive lipase activation in adipocytes (Test Example 3). [Figure 3]3 is a graph showing the effect of the compound of the present invention on cAMP production in adipocytes (Test Example 4). The amount of cAMP produced (%) is expressed as a relative percentage, with the maximum amount of cAMP produced by stimulation with a high concentration (10,000 nM) of isoproterenol (ISO) taken as 100%. [Figure 4] 4 is a graph showing the effect of the compound of the present invention on protein kinase A (PKA) activation in adipocytes (Test Example 5). When PKA in a sample is activated, the substrate contained in the reacted reagent is phosphorylated by the activated PKA and migrates to the cathode (lower side) of electrophoresis. On the other hand, the substrate that is not phosphorylated by PKA migrates to the anode (upper side) of electrophoresis. The stronger the band that migrates to the cathode, the stronger the activation of PKA in the sample. [Figure 5] FIG. 5 is a graph showing the effect of the compound of the present invention on the production of free fatty acids from adipocytes (Test Example 6). [Figure 6] FIG. 6 is a graph showing the synergistic effect of the compound of the present invention and a cAMP analogue (dcAMP) on the production of free fatty acids from adipocytes (Test Example 7). [Figure 7] Figure 7 shows the in vivo effect of the compound of the present invention (Test Example 8). Figure 7A is a graph showing the effect of the compound of the present invention on body weight (Dunnett's test, *p<0.01, vs. control group). Figure 7B is a graph showing the effect of the compound of the present invention on free fatty acid production. Figure 7C is a graph showing the effect of the compound of the present invention on hormone-sensitive lipase activation in adipocytes. Specific Description of the Invention

[0011] <<Definition>> In the present invention, the term "alkyl group" as a whole or part of a group refers to a linear, branched, or cyclic aliphatic hydrocarbon chain. The number of carbon atoms in the alkyl group can be, for example, 1 to 4, 2 to 4, 1 to 3, 2 to 3, 1 to 2, or 1. Examples of the alkyl group include a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a sec-butyl group, a tert-butyl group, a cyclopropyl group, and a cyclobutyl group.

[0012] In the present invention, the term "aliphatic cyclic group" refers to a cyclic aliphatic hydrocarbon group. The aliphatic cyclic group may be carbocyclic or heterocyclic. The carbocyclic aliphatic cyclic group is a 3- to 8-membered (preferably 4- to 7-membered, 5- to 6-membered, or 6-membered) saturated or unsaturated monocyclic carbon ring (aliphatic carbon ring), such as a cyclopropane group, a cyclobutane group, a cyclopentane group, a cyclohexane group, a cycloheptane group, and a cyclooctane group. The heterocyclic aliphatic cyclic group is a 3- to 8-membered (preferably 4- to 7-membered, 5- to 6-membered, or 6-membered) saturated or unsaturated monocyclic hetero ring (aliphatic hetero ring) containing one or more (e.g., 1 to 3, 1 to 2, or 1) heteroatoms selected from oxygen, nitrogen, and sulfur atoms as ring atoms, such as a pyrrolidine group, a piperidine group, a piperazine group, a morpholine group, and a tetrahydrofuran group.

[0013] In the present invention, the term "aromatic ring group" refers to a ring-based compound or a portion thereof having aromatic properties, such as a stable structure containing a cyclic conjugated system with 4n+2 π electrons. The term "aromatic ring group" may be an aromatic hydrocarbon group or an aromatic heterocyclic group. The term "aromatic hydrocarbon group" is a 6- to 10-membered unsaturated carbocyclic ring, and may be a monocyclic aromatic ring group or a bicyclic fused aromatic ring group, such as a phenyl group or a naphthyl group. The term "aromatic heterocyclic group" is a 6- to 10-membered heterocyclic ring containing one or more (e.g., 1 to 3, 1 to 2, or 1) heteroatoms selected from oxygen, nitrogen, and sulfur atoms as ring members, and may be a monocyclic aromatic heterocyclic group or a bicyclic fused aromatic heterocyclic group, such as a furan group, a thiophene group, a pyrrole group, an imidazole group, a pyridine group, a pyrimidine group, a quinoline group, an isoquinoline group, or an indole group.

[0014] In the present invention, the term "alkoxy group" refers to a group in which an alkyl group is bonded to an oxygen atom (-O-), and for example, an alkoxy group having 1 to 4 carbon atoms means that the number of carbon atoms in the alkyl group portion is 1 to 4. Examples of the alkoxy group include a methoxy group and an ethoxy group.

[0015] In the present invention, the term "alkoxycarbonyl group" refers to a group in which an alkoxy group is bonded to a carbonyl group (-C(=O)-) via an oxygen atom (-O-), and for example, an alkoxycarbonyl group having 1 to 4 carbon atoms means that the alkyl group moiety has 1 to 4 carbon atoms. Examples of the alkoxycarbonyl group include a methoxycarbonyl group, an ethoxycarbonyl group, a propoxycarbonyl group, and a butoxycarbonyl group.

[0016] In the present invention, a hydroxyl group represents -OH, and a carboxy group represents -C(=O)-OH.

[0017] In the present invention, the term "halogen atom" means a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom.

[0018] In the present invention, the term "optionally substituted" means that one or more hydrogen atoms on the group or atom to be substituted may be substituted with another group.

[0019] <<Compound>> In formula (I), X and Z each independently represent a carbon atom or a nitrogen atom, but X and Z do not simultaneously represent a nitrogen atom. Preferably, one of X and Z represents a nitrogen atom and the other represents a carbon atom, more preferably, X represents a nitrogen atom and Z represents a carbon atom. When X represents a nitrogen atom, R 5 does not exist, and when Z represents a nitrogen atom, R 6 does not exist.

[0020] In formula (I), --- represents a single bond or a double bond, preferably a double bond. --- represents a single bond, the compound of formula (I) is 2 and R 3 The two carbon atoms to which each of these is bonded may be asymmetric atoms, resulting in stereoisomers (diastereomers). That is, the compounds of formula (I) include cis and trans isomers.

[0021] In formula (I), R 1 The group -R a -R b In R a One of the -(CH)- groups represented by -(CH) may represent -(C=O)-. In this case, -R a and R b is preferably bonded via this -(C=O)-.

[0022] In formula (I), R 1 The group -R a -R b is preferably R a represents -CH2- or -C(=O)-, and R bmay represent an optionally substituted aromatic ring group or an optionally substituted aliphatic carbocyclic group, and a preferred example of the aromatic ring group is phenyl, and a preferred example of the aliphatic carbocyclic group is cyclohexyl.

[0023] In formula (I), R 1 is preferably an alkyl group having 1 to 4 carbon atoms or -R a -R b (In the formula, R a represents -CH2- or -C(=O)-, and R b represents an optionally substituted aromatic ring group or an optionally substituted aliphatic carbocyclic group).

[0024] In formula (I), R 2 and R 3 preferably simultaneously represent the same group, provided that R 2 and R 3 never simultaneously represent a hydrogen atom.

[0025] In formula (I), R 2 and R 3 are R c is an alkoxy group having 1 to 4 carbon atoms, -(C=O)-R c can be expressed as R c preferably represents methoxy or ethoxy.

[0026] In formula (I), R 2 and R 3 are R c -N(-R 11 )(-R 12 ) is a group -(C=O)-R c can be expressed as R 11 and R 12 Preferably, both of represent a hydrogen atom, or one of them represents an alkyl group having 1 to 4 carbon atoms (more preferably methoxy or ethoxy) and the other represents a hydrogen atom.

[0027] In formula (I), R 2 and R 3 are R c-O-(CH2)n-(C=O)-R 13 a group -(C=O)-R c where n is preferably 1 or 2, and R 13 preferably represents methoxy or ethoxy, more preferably n is 1 or 2, and R 13 represents methoxy.

[0028] In formula (I), R 5 is R d is an alkoxy group having 1 to 4 carbon atoms, -(C=O)-R d can be expressed as R d preferably represents methoxy or ethoxy.

[0029] In formula (I), R 5 is R d -N(-R 21 )(-R 22 ) is a group -(C=O)-R d can be expressed as R 21 and R 22 Preferably, both of represent a hydrogen atom, or one of them represents an alkyl group having 1 to 4 carbon atoms (more preferably methoxy or ethoxy) and the other represents a hydrogen atom.

[0030] In formula (I), R 5 is R d -O-(CH2)p-(C=O)-R 23 a group -(C=O)-R d where p is preferably 1 or 2, and R 23 preferably represents methoxy or ethoxy, more preferably p is 1 or 2, and R 23 represents methoxy.

[0031] In formula (I), R 6 is R e is an alkoxy group having 1 to 4 carbon atoms, -(C=O)-R e can be expressed as R e preferably represents methoxy or ethoxy.

[0032] In formula (I), R 6 is R e -N(-R 31 )(-R 32 ) is a group -(C=O)-R e can be expressed as R 31 and R 32 Preferably, both of represent a hydrogen atom, or one of them represents an alkyl group having 1 to 4 carbon atoms (more preferably methoxy or ethoxy) and the other represents a hydrogen atom.

[0033] In formula (I), R 6 is R e -O-(CH2)r-(C=O)-R 33 a group -(C=O)-R e where r is preferably 1 or 2, and R 33 preferably represents methoxy or ethoxy, more preferably r is 1 or 2 and R 33 represents methoxy.

[0034] According to a preferred embodiment of the present invention, in formula (I), one of X and Z represents a nitrogen atom, and the other represents a carbon atom; --- represents a double bond, R 1 is an alkyl group having 1 to 4 carbon atoms or -R a -R b (In the formula, R a represents -CH2- or -C(=O)-, and R b represents an optionally substituted aromatic ring group or an optionally substituted aliphatic carbocyclic group), R 2 and R 3 may be the same or different and represent an alkoxycarbonyl group having 1 to 4 carbon atoms (preferably a methoxycarbonyl group or an ethoxycarbonyl group), R 4 and R 7 represents a hydrogen atom, R 5represents a hydrogen atom or an alkoxycarbonyl group having 1 to 4 carbon atoms, provided that when X represents a nitrogen atom, R 5 does not exist, R 6 represents a hydrogen atom or an alkoxycarbonyl group having 1 to 4 carbon atoms, provided that when Z represents a nitrogen atom, R 6 does not exist A compound or a pharmaceutically acceptable salt or solvate thereof is provided.

[0035] According to a more preferred embodiment of the present invention, in formula (I), X represents a nitrogen atom, Z represents a carbon atom, --- represents a double bond, R 1 is an alkyl group having 1 to 4 carbon atoms or -R a -R b (In the formula, R a represents -CH2- or -C(=O)-, and R b represents an optionally substituted aromatic ring group (preferably a phenyl group) or an optionally substituted aliphatic carbocyclic group (preferably a cyclohexyl group), R 2 and R 3 may be the same or different and represent a methoxycarbonyl group or an ethoxycarbonyl group, preferably R 2 and R 3 Both represent a methoxycarbonyl group or an ethoxycarbonyl group. R 4 and R 7 represents a hydrogen atom, R 5 does not exist, R 6 represents a methoxycarbonyl group or an ethoxycarbonyl group A compound or a pharmaceutically acceptable salt or solvate thereof is provided.

[0036] The compounds of the present invention also include pharmaceutically acceptable salts of the compound of formula (I). Here, "pharmacologically acceptable salts" can be selected from salts described in Berge et al., J. Pharm. Sci. 66:1-19 (1977), etc. Examples of pharmaceutically acceptable salts include, when an acidic group such as a carboxy group is present in the compound of formula (I), salts with alkali metals and alkaline earth metals such as lithium, sodium, potassium, magnesium, and calcium; salts with amines such as ammonia, methylamine, dimethylamine, trimethylamine, dicyclohexylamine, tris(hydroxymethyl)aminomethane, N,N-bis(hydroxyethyl)piperazine, 2-amino-2-methyl-1-propanol, ethanolamine, N-methylglucamine, and L-glucamine; and salts with basic amino acids such as lysine, δ-hydroxylysine, and arginine. When a basic group is present in the compound of formula (I), examples of the salt include salts with mineral acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid; salts with organic acids such as methanesulfonic acid, benzenesulfonic acid, paratoluenesulfonic acid, acetic acid, propionate, tartaric acid, fumaric acid, maleic acid, malic acid, oxalic acid, succinic acid, citric acid, benzoic acid, mandelic acid, cinnamic acid, lactic acid, glycolic acid, glucuronic acid, ascorbic acid, nicotinic acid, and salicylic acid; and salts with acidic amino acids such as aspartic acid and glutamic acid.

[0037] The compounds of the present invention also include solvates of the compounds of formula (I) or their pharmaceutically acceptable salts. Examples of solvates include hydrates and ethanolates.

[0038] <<Production of Compounds>> The compound of formula (I) of the present invention can be prepared according to the following scheme 1. That is, the compound of formula (I) can be prepared by reacting a compound of formula (II) (wherein R1, R4, R5, R6, and R7 have the same meanings as defined in formula (I)) with a compound of formula (III) (wherein R2 and R3 have the same meanings as defined in formula (I)) in an organic solvent (non-polar solvent (toluene, dichloromethane) to polar protic solvent (ethanol, isopropanol), etc.) at room temperature to 120°C for 0.5 to 24 hours.

[0039] [ka]

[0040] The compound of formula (II) can be prepared as described in Synthesis Example 1 in the Examples below.

[0041] The compound of formula (III) is available as a commercially available reagent.

[0042] <<Uses of the compound>> According to the examples described below, the compound of the present invention can promote the decomposition of neutral fat (triacylglycerol) in adipocytes. Therefore, the compound of the present invention can be used to promote the decomposition of neutral fat in adipocytes, and can also be used to treat, prevent, or improve diseases or symptoms that can be treated, prevented, or improved by promoting the decomposition of neutral fat in adipocytes.

[0043] Diseases or symptoms that can be treated, prevented, or improved by promoting triglyceride breakdown in adipocytes include obesity and related diseases or symptoms (Edward T et al., Nat. Metab., 2019, 1(2):189-200; Shannon M et al., Nature Reviews Endocrinology, 2017, 13:633-643). Obesity-related diseases or symptoms include impaired glucose tolerance, dyslipidemia, hypertension, hyperuricemia, gout, coronary artery disease, cerebral infarction, fatty liver, menstrual disorders, pregnancy complications, sleep apnea syndrome, obesity-hypoventilation syndrome, orthopedic diseases, and obesity-related kidney disease (Obesity Diagnostic Criteria 2011, Japan Society for the Study of Obesity).

[0044] According to the examples below, the compound of the present invention can synergistically enhance the neutral fat decomposition promoting effect of cAMP analogues when used in adipocytes in combination with cAMP analogues.Therefore, the compound of the present invention can preferably be used in combination with a cAMP production promoter in adipocytes.Examples of the cAMP production promoter in adipocytes include isoproterenol, dcAMP and forskolin.

[0045] As an anti-obesity drug that induces the production of cAMP in adipocytes and decomposes neutral fat, cAMP production promoters in adipocytes are known, but the neutral fat decomposition effect of compounds that induce cAMP production, such as isoproterenol, dcAMP and forskolin, disappears 24 hours after the introduction of these compounds into adipocytes, and is transient.In contrast, the compound of the present invention continues to promote neutral fat decomposition without attenuation even after 24 hours and 72 hours after the introduction of these compounds into adipocytes.In other words, the compound of the present invention is advantageous in that it can be expected to continuously decompose neutral fat in mast cells.

[0046] Since the compounds of the present invention can promote the breakdown of neutral fat in adipocytes without cAMP production, it is believed that the compounds of the present invention can promote neutral fat breakdown through a pathway that does not involve β-adrenergic receptor stimulation (activation of protein kinase A and activation of hormone-sensitive lipase). Therefore, the compounds of the present invention can be used for the treatment, prevention, or amelioration of obesity, in which neutral fat accumulates in adipocytes due to decreased β-adrenergic receptor (β2 or β3 receptor) stimulation, as well as diseases and symptoms associated therewith.

[0047] Since the compounds of the present invention have the effect of promoting triglyceride breakdown in adipocytes and also have the effect of reducing body weight, they can be administered to subjects at risk of developing obesity or a disease or symptom associated therewith, thereby reducing the risk of developing obesity or a disease or symptom associated therewith. Here, "subjects at risk of developing obesity or a disease or symptom associated therewith" means subjects who do not have any subjective symptoms of obesity or the like, but who are at risk of developing obesity or the like in the future. Furthermore, "reducing the risk of developing obesity or a disease or symptom associated therewith" means reducing the probability of developing obesity or the like. That is, according to another aspect of the present invention, there are provided an agent for reducing the risk of developing obesity or a disease or symptom associated therewith, and a composition for reducing the risk of developing obesity or a disease or symptom associated therewith, which comprise the compound of the present invention as an active ingredient.

[0048] <<Medicines and Treatment Methods>> The present invention provides pharmaceutical compositions and therapeutic preparations comprising the compounds of the present invention as active ingredients. The route of administration of the compounds of the present invention is not particularly limited and may be oral or parenteral (e.g., intravenous or subcutaneous). Oral preparations include, for example, tablets, capsules, granules, powders, pills, lozenges, chewable tablets, syrups, liquids, emulsions, and suspensions. Parenteral preparations include, for example, injections, suppositories, inhalants, and transdermal absorbents. These preparations can be formulated using pharmaceutically acceptable carriers by methods commonly used in the art (e.g., known methods described in the General Provisions for Preparations of the Japanese Pharmacopoeia, 18th Edition). Examples of pharmaceutically acceptable carriers include excipients, binders, diluents, lubricants, additives, flavorings, buffers, thickeners, colorants, stabilizers, emulsifiers, dispersants, suspending agents, and preservatives.

[0049] The dosage of the compound of the present invention depends on the sex, age, and weight of the subject, symptoms, intake time, dosage form, administration route, and other combined drugs, and can be appropriately determined by a physician's judgment in some cases. When the compound of the present invention is administered for the purpose of promoting triglyceride breakdown in adipocytes or treating, preventing, or ameliorating diseases or symptoms that can be treated, prevented, or ameliorated by promoting triglyceride breakdown in adipocytes, the daily dosage for an adult can be, for example, in the range of 0.01 to 1000 mg when administered orally, and can be, for example, in the range of 0.001 to 100 mg when administered parenterally, such as by intravenous injection, but is not limited thereto. The compound of the present invention can be administered not only to humans in need thereof, but also to non-human mammals (e.g., mice, rats, rabbits, dogs, cats, cows, horses, pigs, sheep, goats, and monkeys).

[0050] According to another aspect of the present invention, there is provided a method for promoting neutral fat breakdown in adipocytes, comprising administering an effective amount of the compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or a composition comprising the same, to a subject in need thereof. According to another aspect of the present invention, there is also provided a method for treating, preventing, or ameliorating a disease or symptom that can be treated, prevented, or ameliorated by promoting neutral fat breakdown in adipocytes, comprising administering an effective amount of the compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or a composition comprising the same, to a subject in need thereof. According to another aspect of the present invention, there is also provided a method for reducing the risk of developing obesity or a disease or symptom associated therewith, comprising administering an effective amount of the compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or a composition comprising the same, to a subject in need thereof. The method of the present invention can be carried out according to the description of the compounds and medicaments of the present invention.

[0051] Another aspect of the present invention provides use of the compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or a composition comprising the same, for the manufacture of an agent for promoting neutral lipolysis in adipocytes, as an agent for promoting neutral lipolysis in adipocytes, or in a method for promoting neutral lipolysis in adipocytes. Another aspect of the present invention provides use of the compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or a composition comprising the same, for the manufacture of an agent for treating, preventing, and / or improving a disease or symptom that can be treated, prevented, or improved by promoting neutral lipolysis in adipocytes, as an agent for treating, preventing, and / or improving a disease or symptom that can be treated, prevented, or improved by promoting neutral lipolysis in adipocytes, or in a method for treating, preventing, or improving a disease or symptom that can be treated, prevented, or improved by promoting neutral lipolysis in adipocytes. According to another aspect of the present invention, there is further provided use of the compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or a composition containing the same, for the manufacture of an agent for reducing the risk of developing obesity or a disease or symptom associated therewith, as an agent for reducing the risk of developing obesity or a disease or symptom associated therewith, or in a method for reducing the risk of developing obesity or a disease or symptom associated therewith. The use of the present invention can be carried out according to the description related to the compound and medicament of the present invention.

[0052] According to yet another aspect of the present invention, there is provided a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or a composition comprising the same, for use in the method of the present invention. That is, according to another aspect of the present invention, there is provided a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or a composition comprising the same, for use in promoting triglyceride breakdown in adipocytes. According to another aspect of the present invention, there is also provided a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or a composition comprising the same, for use in the treatment, prevention, and / or amelioration of diseases or symptoms that can be treated, prevented, or ameliorated by promoting triglyceride breakdown in adipocytes. According to another aspect of the present invention, there is also provided a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or a composition comprising the same, for use in reducing the risk of developing obesity or a disease or symptom associated therewith. The above invention can be carried out according to the description of the compounds and medicaments of the present invention. [Example]

[0053] The present invention will be described in more detail based on the following examples, but the present invention is not limited to these examples.

[0054] Synthesis Example 1: Synthesis of ethyl 2-benzyl-2H-pyrrolo[3,4-c]pyridine-6-carboxylate (Lipid Release Inducing Stimulant 023, also simply referred to as "LIS023") [ka]

[0055] Compound LIS023 was prepared as follows. (1) Preparation of N-benzyl-3,4-dicyanopyrrole [ka]

[0056] Fumaronitrile (1199.6 mg, 15.3657 mmol) and paratoluenesulfonylmethyl isocyanate (TosMIC) (3000.9 mg, 15.3657 mmol) in 100 mL of THF were added with 60% sodium hydride (737.2 mg, 18.4388 mmol) dispersed in mineral oil at 0 °C. The mixture was then heated to 40 °C and stirred for 2 hours. Benzyl bromide (2.75 mL, 3932.5 mg, 22.9917 mmol) and 60% sodium hydride (740.2 mg, 18.5050 mmol) dispersed in mineral oil were added at 0 °C, and the mixture was then heated to 40 °C and reacted for 4 hours. The reaction mixture was concentrated, and 200 mL of 1 mol / L hydrochloric acid was added to the residue, followed by extraction with 200 mL of ethyl acetate three times. The organic layer was washed with 200 mL of 1 mol / L hydrochloric acid, then with 200 mL of water twice, and finally with 200 mL of saturated brine. The organic layer was dried over sodium sulfate and concentrated. The residue was purified by silica gel chromatography (hexane:acetone = 5:1 → 3:1 → 1:1) to give N-benzyl-3,4-dicyanopyrrole as a yellow solid (2060.1 mg, 9.5408 mmol, 68% yield).

[0057] 1 H NMR (CDCl3, 400 MHz): δ (ppm) = 7.430-7.400(m, 3H), 7.190-7.160(m, 2H),7.180 (s, 2H) 5.100 (s, 2H); 13 C NMR (CDCl3, 100 MHz): δ (ppm) = 133.70,129.49,129.36,129.27,127.81, 112.58, 91.53, 54.88; HRMS (ESI-TOF, [M+H] + ): Calculated value (C 13 H 10 N3), 208.0869. Measurement, 208.0868.; Elemental analysis Calculated value (C 13 H9N3): N, 20.28; C, 75.35; H, 4.38. Found: N, 20.18; C, 75.21; H, 4.59; mp 154.0-155.0°C (needles, recrystallized from CH2Cl2 / hexane).

[0058] (2) Preparation of N-benzyl-3,4-dialdehydepyrrole [ka]

[0059] To the N-benzyl-3,4-dicyanopyrrole (1543.8 mg, 7.4497 mmol) obtained in (1) above in 100 mL of methylene chloride, 15 mL (15 mmol) of a 1 mol / L toluene solution of diisobutylaluminum hydride was added over 5 minutes at 0°C under an argon atmosphere, and the mixture was stirred for 1 hour. Approximately 30 g of sodium sulfate octahydrate was then added to quench the reaction, followed by filtration through Celite. The filtrate was concentrated and purified by silica gel chromatography (hexane:acetone = 1:1) to obtain N-benzyl-3,4-dialdehydepyrrole as a colorless solid. This was further recrystallized from ethyl acetate / hexane (1246.4 mg, 5.8476 mmol, 78% yield).

[0060] 1 H NMR (CDCl3, 400 MHz): δ (ppm) = 10.148 (s, 2H), 7.408-7.375(m, 3H), 7.361(s, 2H), 7.220-7.190 (m, 2H), 5.119 (s, 2H); 13 C NMR (CDCl3, 100 MHz): δ (ppm) = 186.33, 134.55, 129.74, 129.34, 129.01, 127.09, 125.40, 54.58; HRMS (ESI-TOF, [M+Na] + ): Calculated value [C 13 H 11 O2NNa] + , 236.0682. Measurement:236.0682; Elemental analysis Calculated value (C 13 H 11 O2N): N, 6.57; C, 73.23; H, 5.20. Measured values: N, 6.71; C, 73.08; H, 5.31. Melting point: 68.0-69.0°C (needles, recrystallized from ethyl acetate / hexane)

[0061] (3) Preparation of 2-benzyl-2H-pyrrolo[3,4-c]pyridine-6-ethylcarboxylate [ka]

[0062] To the N-benzyl-3,4-dialdehydepyrrole (1010.8 mg, 4.7404 mmol) obtained in (2) above in 90 mL of ethanol, diethylamine (1.47 mL, 1040.1 mg, 14.2212 mmol) and glycine ethyl ester hydrochloride (1323.3 mg, 9.4808 mmol) were added at room temperature, and the reaction mixture was refluxed for 1 hour. The mixture was then cooled to room temperature and concentrated. The residue was dissolved in 90 mL of water and extracted three times with 90 mL of ethyl acetate. The organic layer was washed twice with 90 mL of water and with 90 mL of saturated brine, and then dried over sodium sulfate. The solvent was removed, and the residue was purified by silica gel chromatography (acetone / hexane = 1:1) to give 2-benzyl-2H-pyrrolo[3,4-c]pyridine-6-ethylcarboxylate as a yellow solid (1200.3 mg, 4.2818 mmol, 90% yield).

[0063] 1 H-NMR (400 MHz, CDCl3): δ (ppm) = 9.06 (1H, s), 8.36 (1H, s), 7.39 - 7.32 (5H, m), 7.19 - 7.17 (2H, m), 5.42 (2H, s), 4.46 (2H, q, J = 7.2 Hz), 1.44 (3H, t, J = 7.2 Hz); 13 C-NMR (100 MHz, CDCl3): δ (ppm) = 166.82, 146.24, 136.23, 135.67, 129.12, 128.65, 127.58, 124.43, 121.98, 118.32, 114.25, 114.09, 61.22, 55.45, 14.51; HRMS (ESI-TOF, [M+Na] + ): Calculated value [C 17 H 16 N2NaO2 + ]: 303.1104. Measurement: 303.1068.; Elemental analysis Calculated value [C 17 H 16 N2O2]; C, 72.84; H, 5.75; N, 9.99. Measured values: C, 72.64; H, 5.82; N, 9.83. Melting point: 109-110 o C (recrystallized from ethyl acetate / hexane).

[0064] Synthesis Example 2: Synthesis of 3-ethyl 6,7-dimethyl(5R,8S)-9-benzyl-5,8-dihydro-5,8-epiminoisoquinoline-3,6,7-tricarboxylate (fat release-inducing stimulating compound 001, also simply referred to as "LIS001") [ka]

[0065] To a stirred solution of LIS023 (200.0 mg, 0.7135 mmol) in 3 mL of toluene, dimethyl acetylenedicarboxylate (132 μL, 152.6 mg, 1.0703 mmol) was added. After 4 h, the whole was directly subjected to column chromatography (hexane / AcOEt = 6:1, 4:1, 2:1, 1:1, 1:2) to give LIS001 (257.0 mg, 0.6084 mmol, 85%) as a yellow amorphous solid.

[0066] 1 H NMR (400 MHz, CDCl3):δ(ppm) = 8.687 (s, 1H), 8.150 (s, 1H), 7.350-7.280 (m 3H), 7.270-6.900 (m 2H), 5.054 (s, 1H), 5.014 (s, 1H), 4.514-4.435 (m, 2H), 3.802 (s, 3H), 3.800 (s, 3H), 3.580 (s, 2H), 1.434 (t, J = 7.2 Hz, 3H). 13C NMR (100 MHz, CDCl3):δ(ppm) = 164.29, 162.78, 157.64, 151.54, 148.90,148.07, 146.29, 145.26, 141.55, 136.08, 128.47, 128.15, 127.23, 118.93,74.08, 71.96, 70.55, 61.46, 52.92, 52.56, 52.01, 13.81. HRMS (ESI-TOF, [M+Na] + ):Analysis result:C 23 H 22 N2NaO6 + Calculated value: 445.1370. Measured value: 445.1373. Analysis result: C 23 H 22 N2O6, calculated: + 0.2 acetone + 0.2 H2O + 0.1 hexane; C, 64.21; H, 5.57; N, 6.19. Found: C, 64.35; H, 5.97; N, 6.13.

[0067] Synthesis Example 3: Synthesis of trimethyl(5R,8S)-9-benzyl-5,8-dihydro-5,8-epiminoisoquinoline-3,6,7-tricarboxylate (fat release-inducing stimulating compound 016, also simply referred to as "LIS016") [ka]

[0068] To a stirred solution of the LIS023 derivative (methyl ester) (607.6 mg, 2.2816 mmol) in 5 mL of toluene, dimethyl acetylenedicarboxylate (421 μL, 486.4 mg, 3.4224 mmol) was added. After 12 h, the whole was directly subjected to column chromatography (hexane / acetone = 6:1, 4:1, 2:1, 1:1) to give LIS016 (619.8 mg, 1.5102 mmol, 66%) as a yellow amorphous solid.

[0069] 1H NMR (400 MHz, CDCl3): δ(ppm) = 8.699 (s, 1H), 8.180 (s, 1H), 7.354-7.327 (m 3H), 7.280-7.233 (m 2H), 5.085 (s, 1H), 5.042 (s, 1H), 4.024 (s, 3H), 3.827 (s, 3H), 3.825 (s, 3H), 3.612 (brs, 2H), 3.800 (s, 3H), 3.580 (s, 2H), 1.434 (t, J = 7.2 Hz, 3H). 13 C NMR (100 MHz, CDCl3): δ(ppm) = 165.11, 163.04, 157.97, 149.12, 148.37, 146.25,145.68,141.80,136.27,128.75,128.47, 128.46, 127.55, 119.16, 72.29, 70.85, 52.88, 52.72, 52.34, 52.32. HRMS (ESI-TOF, [M+Na] + Analysis results: C 22 H 20 N2NaO6 + , Calculated value: 431.12136. Measured value: 431.12174. Analysis result: C 22 H 20 N2O6, calculated value: + 0.7H2O; C, 62.76; H, 5.12; N, 6.65. Measured value: C, 62.57; H, 4.83; N, 6.55.

[0070] Synthesis Example 4: Synthesis of trimethyl(5S,6S,7R,8R)-9-benzyl-5,6,7,8-tetrahydro-5,8-epiminoisoquinoline-3,6,7-tricarboxylate (racemic form) (fat release-inducing stimulating compound 017, also simply referred to as "LIS017")

change

[0071] To a stirred solution of the LIS023 derivative (methyl ester) (500.3 mg, 1.8787 mmol) in 4 mL of dry toluene, maleic anhydride (552.7 mg, 5.6363 mmol) and hydroquinone (10.3 mg, 0.0939 mmol) were added. The whole was heated to 67 °C in a microwave reactor. After 3 h, the reaction mixture was allowed to cool to room temperature. The whole was directly subjected to column chromatography (hexane / AcOEt = 1:1, 1:2, 1:3) to obtain the Diels-Alder adduct (502.3 mg) as a brown amorphous solid. The intermediate was immediately dissolved in 5 mL of methanol, and 4 mol / L HCl in 0.5 mL of AcOEt was added. After 2 h, the whole was evaporated and dried under vacuum. The resulting residue was dissolved in 5 mL of dry methanol, and 0.6 mol / L TMSCHN2 in 2.5 mL of hexane was added. After stirring for 20 min, the whole was evaporated, and the resulting residue was purified by column chromatography (hexane / acetone = 6:1, 4:1, 2:1, 1:1) to give LIS017 (240.5 mg, 0.5860 mmol, 43%) as a pale yellow amorphous solid.

[0072] 1 H NMR (400 MHz, CDCl3):δ(ppm) = 8.671 (s, 1H), 8.108 (s, 1H), 7.310-7.232 (m 5H), 4.647 (s, 1H), 4.606 (s, 1H), 4.022 (s, 3H), 3.735 (s, 6H), 3.695 (s, 2H), 2.823 (d, J = 9.2 Hz,1H), 2.762 (d, J = 9.6 Hz,1H). 13 C NMR (100 MHz, CDCl3):δ(ppm) = 170.89, 170.83, 165.46, 155.05, 147.53, 143.40, 142.95, 137.90, 128.21, 128.14, 127.17, 119.2767.17, 67.01, 65.66, 52.14, 47.43. HRMS (ESI-TOF, [M+Na] + ): Analysis result:C 22 H22 N2NaO6 + Calculated value: 433.13701. Measured value: 433.13751. Analysis result: C 22 H 22 N2O6, calculated: + 0.3 acetone; C, 64.29; H, 5.61; N, 6.55. Found: C, 64.61; H, 5.93; N, 6.37.

[0073] Synthesis Example 5: Synthesis of 3-ethyl 6,7-dimethyl(5R,8S)-9-benzoyl-5,8-dihydro-5,8-epiminoisoquinoline-3,6,7-tricarboxylate (fat release-inducing stimulating compound 018, also simply referred to as "LIS018") [ka]

[0074] To a stirred solution of LIS001 (2.6367 g, 6.2416 mmol) in 40 mL of dioxane and 20 mL of HO, NBS (4.4435 g, 24.9666 mmol) and BPO (15.1 mg, 0.0624 mmol) were added. The reaction mixture was heated to 70 °C and allowed to reach that temperature. The reaction mixture was allowed to stand at room temperature. After 24 h, the whole was evaporated and dried under vacuum. The resulting residue, which was almost solidified, was dissolved in 40 mL of dry THF. This solution was added to triethylamine (8.9 mL, 62.4164 mmol) and benzoyl chloride (2.9 mL, 24.9666 mmol). The whole was stirred for 16 h. After completion of the reaction, the whole was poured into 100 mL of saturated aqueous NaHCO3 solution and extracted three times with 100 mL of EtOAc. The combined organic layers were washed with 100 mL of HO, 100 mL of saturated aqueous NH4Cl, and 100 mL of brine, and dried over Na2SO4. The solvent was evaporated, and the residue was purified by column chromatography (hexane / AcOEt = 6:1, 4:1, 2:1, 1:1, 1:2, 1:3) to give LIS018 (2.1164 g, 4.8495 mmol, 78%) as an orange amorphous solid.

[0075] 1H NMR (400 MHz, CDCl3): δ (ppm) = 8.682 (s, 1H), 8.161 (s, 1H), 7.448 (m 3H), 6.193 (brs 2H), 6.010 (brs, 1H), 4.388 (quartet, J = 6.8 Hz, 2H), 3.724 (brs, 6H), 1.339 (t, J = 6.8 Hz, 3H). 13 C NMR (100 MHz, CDCl3): δ (ppm) = 168.95, 164.15, 161.61, 157.02, 150,75, 148.90, 144.41, 140.90, 132.06, 131.96, 128.57, 127.88, 118.08, 69,54, 68.31, 66.31, 65.06, 61.87, 52.52, 13.99. HRMS (ESI-TOF, [M+Na] + Analysis results: C 23 H 20 N2NaO7 + Calculated value: 459.11627. Measured value: 433.13751459.11631. Analytical result: C 23 H 20 N2O7, Calculated value: + 0.3 AcOEt + 0.2 H2O; C, 62.80; H, 4.88; N, 6.05. Determined value: C, 62.46; H, 4.91; N, 6.16.

[0076] Test Example 1: Preparation of mature adipocytes Mouse fibroblast 3T3-L1 cells were seeded onto a dish and cultured until confluent. To induce differentiation of confluent 3T3-L1 cells into adipocytes, the medium was replaced with 10% fetal bovine serum, Dulbecco's modified Eagle's medium (high glucose 4.5 g / L) (hereinafter referred to as "DMEM"), 500 μM isobutylmethylxanthine (Sigma, hereinafter referred to as "IBMX"), 5 μg / mL insulin, and 250 nM dexamethasone (DEX) (Day 0). Two days later (Day 2), the medium was replaced with DMEM medium (10% fetal bovine serum, 5 μg / mL insulin). Six days after the medium replacement (Day 8), differentiation into mature adipocytes was completed. Mature adipocytes from Day 8 or later were used as adipocytes in the examples described below.

[0077] Test Example 2: Neutral fat decomposition promoting effect of compound LIS (1) (1) Experimental procedure Mature adipocytes prepared in Test Example 1 were used. The medium was replaced with DMEM medium (2% bovine serum albumin (Wako, hereinafter the same), various agonists added). The agonists used were compound LIS001 (100 μM), isoproterenol (β-adrenergic receptor agonist, sometimes referred to as "ISO" herein) (100 nM and 100 μM), dibutyryl cAMP (cAMP analog, sometimes referred to as "dcAMP" herein) (300 μM), or forskolin (adenylate cyclase activator) (1 μM). A control group (no agonist added) was also prepared. 3, 24, 48, and 72 hours after agonist administration, a portion of the cell supernatant was collected, and the free fatty acid concentration in the supernatant was measured using a non-esterified fatty acid kit (NEFA C-Test Wako, Wako Pure Chemical Industries, Ltd.). Specifically, the supernatant of the collected cells was reacted with the reagents in the kit, and the absorbance (wavelength 550 nm) was measured using a spectrophotometer. The concentration (μM) of free fatty acids in each sample was calculated based on the absorbance of the free fatty acid (palmitic acid) whose concentration was known and was included in the kit.

[0078] (2) Results The results are shown in FIG.

[0079] Addition of compound LIS001 to adipocytes was shown to induce the production of free fatty acids from the adipocytes. Furthermore, the free fatty acid production effect of compound LIS001 was shown to be sustained, unlike the transient free fatty acid production effects of agonists (isoproterenol), forskolin, and dcAMP, which induce cAMP production through β-adrenergic receptor stimulation. Specifically, free fatty acid production was observed 30 minutes to 1 hour after administration of compound LIS001, and was confirmed to persist unabated even at 24 and 72 hours after administration. These findings confirm that compound LIS001 has the ability to sustainably promote the breakdown of neutral fats in adipocytes.

[0080] Test Example 3: Hormone-sensitive lipase activation effect of compound LIS (1) Experimental procedure Mature adipocytes prepared in Test Example 1 were used. The medium was replaced with DMEM medium (2% bovine serum albumin, various agonists added). The agonists used were compound LIS001 (300 μM), compound LIS016 (300 μM), compound LIS017 (300 μM), and isoproterenol (100 nM and 1 μM). The control group was a group to which no agonist was added (non-addition). One hour and 24 hours after agonist administration, adipocytes were harvested using SDS (composition: 0.5 M Tris-HCl (pH 6.8) (prepared with Sigma Tris and Wako 1 M HCl), 10% SDS (Wako), 50% glycerol (Wako), and β-mercaptoethanol (2-mercaptoethanol, min. 98%, Sigma). The hormone-sensitive lipase (HSL) activity of the harvested cells was assessed. HSL is an enzyme responsible for lipolysis, and its activity is positively regulated by phosphorylation of Ser563 and Ser660 (generally enhanced via cAMP-PKA stimulation) and negatively regulated by phosphorylation of Ser565 (Antonis D. Lampidonis et al., Gene, 2011, 477: 1-11). HSL activity was assessed by Western blotting according to standard methods. Specifically, the collected samples were boiled and subjected to polyacrylamide gel electrophoresis (SDS-PAGE) to detect intracellular HSL phosphorylated proteins. The primary antibodies used were anti-phosphorylated HSL Ser563 antibody (Phospho-HSL (S563) Rabbit Ab, Cell Signaling Technology, Inc.; the same applies hereinafter), anti-phosphorylated HSL Ser565 antibody (Phospho-HSL (S565) Rabbit Ab, Cell Signaling Technology, Inc.), and anti-HSL antibody (HSL Rabbit Ab, Cell Signaling Technology, Inc.). The intensity of phosphorylation of HSL Ser563 by compound LIS (band intensity) was assessed using the anti-phosphorylated HSL Ser563 antibody.To verify that differences in the intensity of phosphorylation of HSL Ser563 were not due to differences in the quantity of cells collected, the amount of HSL protein in each sample was assessed using an anti-HSL antibody. The intensity of dephosphorylation of HSL Ser565 by the compound LIS was assessed using an anti-phosphorylated HSL Ser565 antibody. High HSL activity resulted in a weaker band detected by the anti-phosphorylated HSL Ser565 antibody, due to the enhanced dephosphorylation of Ser565.

[0081] (2) Results The results are shown in FIG.

[0082] In adipocytes treated with compounds LIS001 or LIS016, increased levels of hormone-sensitive lipase phosphorylated at Ser563 and decreased levels of hormone-sensitive lipase phosphorylated at Ser565 were observed compared to control cells (no treatment). These results suggest that compounds LIS001 and LIS016 activate hormone-sensitive lipase by enhancing the phosphorylation of Ser563 and strongly inhibiting the phosphorylation of Ser565. Furthermore, the hormone-sensitive lipase activation effect of compounds LIS001 and LIS016 was sustained, unlike the transient activation of hormone-sensitive lipase induced by isoproterenol (ISO)-stimulating β-adrenergic receptors, which disappeared within 24 hours after administration. Furthermore, these results and those of Experiment 4 (described below) suggest that LIS016 activates hormone-sensitive lipase, in part, via PKA activation, independent of cAMP. In adipocytes treated with compound LIS017, no increase in the Ser563 phosphorylated protein or decrease in the Ser565 phosphorylated protein of hormone-sensitive lipase was observed. These results demonstrate that the addition of compounds LIS001 and LIS016 to adipocytes strongly and sustainably activates hormone-sensitive lipase in adipocytes.

[0083] Test Example 4: Effect of Compound LIS on cAMP Production (1) Experimental procedure Mature adipocytes prepared in Test Example 1 were used. The medium was replaced with cAMP assay buffer (1x HBSS, prepared by mixing the following: KCl: Wako, KH2PO4: Sigma, NsCl: Sigma, Na2HPO4: Sigma, D-Glucose: Nacalai Tesque), 1 M HEPES (Sigma), 7.5% bovine serum albumin, 500 mM IBMX, and various agonists). The agonists used were compound LIS016 (100,000 nM) and ISO (0, 0.1, 1, 10, 100, 1,000, and 10,000 nM). Thirty minutes after agonist administration, 10% Triton X was added to the cells and incubated for 30 minutes. A portion of the extracellular supernatant was then collected, and the cAMP levels in the supernatant were assessed using the LANCE Ultra cAMP Kit (PerkinElmer). Specifically, the supernatant was reacted with the reagents provided in the kit, and the amount of cAMP produced (TR-FRET signal intensity, wavelength 665 nm) was measured using a multiplate reader (EnSpire, PerkinElmer). Note that the higher the cAMP concentration in the supernatant, the lower the TR-FRET signal, so the amount of cAMP produced was expressed as the degree of decrease in the TR-FRET signal from the baseline.

[0084] (2) Results The results are shown in FIG.

[0085] It was confirmed that the amount of cAMP produced by adipocytes increased in a concentration-dependent manner when β-adrenergic receptors were stimulated with isoproterenol (ISO), whereas administration of the compound LIS016 did not produce cAMP. Therefore, it was confirmed that stimulation by the compound LIS016 is different from stimulation of β-adrenergic receptors (β2 and β3 receptors), which is accompanied by cAMP production, and that it does not enhance cAMP production induced by stimulation of β-adrenergic receptors (β2 and β3 receptors).

[0086] Test Example 5: Protein kinase A activation effect of compound LIS (1) Experimental procedure Mature adipocytes prepared in Test Example 1 were used. The medium was replaced with DMEM medium (containing a PKA inhibitor). 30 μM H-89 was used as the PKA inhibitor. One hour after H-89 administration, various agonists were added to the DMEM medium. The agonist used was compound LIS001 (100 μM), and as a positive control, 6-benzoyl-cAMP (6-Bnz-cAMP: a cAMP analog, Life Science Institute) (300 μM). 6-Bnz-cAMP activates protein kinase A (sometimes referred to as "PKA" herein) (Johannes L. Bos., Trends Biochem. Sci., 2006, 31:680-686). A control group (no agonist added) was also prepared. One and a half hours after agonist administration, adipocytes were harvested using PKA assay buffer (1 M tris HCl, EDTA, EGTA, β-mercaptoethanol, protease inhibitors). PKA activity in the harvested cells was assessed using the PepTag Assay for Non-Redioactive Detection of Protein Kinase C or cAMP-Dependent Protein Kinase (Promega). Specifically, cells harvested in PKA assay buffer were disrupted on ice using a Dounce homogenizer and centrifuged at 14,000 g to separate intracellular components into liquid and solid phases. The liquid phase was harvested, and a portion of the liquid phase was reacted with the reagents provided in the kit, then applied to an agarose gel for electrophoresis.

[0087] (2) Results The results are shown in FIG.

[0088] It was shown that PKA was activated in adipocytes treated with compound LIS001 compared to the control group (no addition). It was also confirmed that PKA was activated in adipocytes treated with the positive control 6-Bz cAMP. On the other hand, it was shown that PKA activation by compound LIS001 or 6-Bz cAMP was inhibited by pre-administration of the PKA inhibitor H-89. These results, along with those of Test Example 4, suggest that PKA activation by compound LIS001 is cAMP-independent.

[0089] Test Example 6: Neutral fat decomposition promoting effect of compound LIS (2) (1) Experimental procedure Mature adipocytes prepared in Test Example 1 were used. The medium was replaced with DMEM medium (2% bovine serum albumin, various agonists added). 100 μM of each of the LIS compounds LIS001, LIS016, LIS017, and LIS018 was used alone as the agonist. A control group (no agonist added) was also prepared. The free fatty acid concentration of each sample was calculated in the same manner as in Test Example 2(1), except that a portion of the cell supernatant was collected 24 hours after agonist administration.

[0090] (2) Results The results are shown in FIG.

[0091] It was confirmed that the ability to produce free fatty acids from adipocytes differed depending on the compound LIS, i.e., the degree of promotion of neutral fat breakdown in adipocytes differed depending on the compound LIS.

[0092] Test Example 7: Synergistic effect of compound LIS and cAMP analogues on promoting neutral fat decomposition From the results of Test Example 6, compounds LIS017 and LIS018 alone were not confirmed to have the effect of promoting neutral fat decomposition. These compounds LIS were mainly evaluated for their ability to produce free fatty acids due to the synergistic effect of compound LIS and dcAMP.

[0093] (1) Experimental procedure Mature adipocytes prepared in Test Example 1 were used. The medium was replaced with DMEM medium (supplemented with 2% bovine serum albumin, various agonists, and dcAMP). The agonists used were LIS017 and LIS018 at 200 μM each, and dcAMP at 300 μM. Control groups included a group without agonist or dcAMP (non-added) and a group with dcAMP only (dcAMP only). The free fatty acid concentration of each sample was calculated in the same manner as in Example 2(1), except that a portion of the cell supernatant was collected 4 hours after agonist administration. The free fatty acid concentration of each sample reflected the free fatty acid production ability of compound LIS and dcAMP. The stronger the free fatty acid production ability of compound LIS and dcAMP, the stronger the synergistic effect of compound LIS and dcAMP was judged to be.

[0094] (2) Results The results are shown in FIG.

[0095] In Test Example 6, compounds LIS017 and LIS018, which were not confirmed to have a neutral fat degradation promoting effect when used alone, were confirmed to synergistically enhance the production of free fatty acids from adipocytes by cAMP analogs when administered in combination with a cAMP analog. This suggests that compounds LIS017 and LIS018 can synergistically enhance the neutral fat degradation promoting effect of cAMP analogs when administered in combination with a cAMP analog.

[0096] Test Example 8: In vivo neutral fat degradation promoting effect of compound LIS (1) Experimental procedure Sixteen male 16-week-old C57 / BL6 mice were weighed. Each weighed approximately 30 g. They were then anesthetized using a triple-dose anesthesia mixture (medetomidine hydrochloride 0.3 mg / kg, midazolam 4 mg / kg, butorphanol tartrate 5 mg / kg). LIS001 (30 mg / 0.03 mL DMSO) was diluted with 3 mL of olive oil and administered intraperitoneally at doses of 0, 10, 30, or 100 mg / kg body weight. After 24 and 72 hours of administration, the mice were weighed and sacrificed. Blood was collected from the heart and the caudal vena cava. The blood was allowed to stand for 30 minutes, then centrifuged at 4°C and 1000 x g for 20 minutes. The free fatty acid concentration (μM) in the supernatant (serum) was calculated as described in Experimental Example 2(1). In addition, adipose tissue was extracted from the epididymis, and the activity of hormone-sensitive lipase (HSL) in the adipocytes was evaluated by Western blotting using anti-phosphorylated HSL Ser563 antibody in the same manner as in Test Example 3(1).

[0097] (2) Results The results are shown in FIG.

[0098] Figure 7A shows that the group administered 30 mg / kg of compound LIS001 experienced a decrease in body weight 72 hours after administration compared to before administration. The group administered 100 mg / kg of compound LIS001 experienced a significant decrease in body weight before and after administration compared to the control group (no administration). Figure 7B shows that administration of compound LIS001 in vivo increased serum free fatty acid levels compared to the control group (no administration), and this effect persisted even 72 hours after administration. These results confirm that compound LIS001 has a sustained effect on triglyceride degradation in vivo. Figure 7C shows that the group administered compound LIS001 exhibited an increase in the amount of phosphorylated protein at Ser563 of hormone-sensitive lipase compared to the control group (no administration). These results indicate that compound LIS001 activates hormone-sensitive lipase in vivo by enhancing the phosphorylation of Ser563. Furthermore, it was shown that the hormone-sensitive lipase activation effect of compound LIS001 persisted for 72 hours after administration. These results indicate that in vivo administration of compound LIS001 potently and sustainably activates hormone-sensitive lipase in adipocytes. These results suggest that in vivo administration of compound LIS001 sustains hormone-sensitive lipase activation in adipocytes, promotes triglyceride breakdown, increases serum free fatty acid levels, and reduces body weight.

Claims

1. A compound of the following formula (I) or a pharmaceutically acceptable salt or solvate thereof: 【Chemistry 1】 (In the above formula, X and Z each represent a carbon atom or a nitrogen atom, and when X represents a nitrogen atom, R ５ does not exist, and when Z represents a nitrogen atom, R ６ does not exist, and both X and Z do not represent nitrogen atoms. --- represents a single or double bond, R １ represents a hydrogen atom, an alkyl group having 1 to 4 carbon atoms, an alkylcarbonyl group having 1 to 4 carbon atoms, or -R ａ -R ｂ (In the formula, R ａ Ha-(CH ２ )m- (wherein m is an integer of 1 to 3), and -(CH ２ ) m- one of -(CH ２ )- may represent -(C=O)-, and R ｂ represents an aromatic ring group or an aliphatic ring group, and the aromatic ring group and the aliphatic ring group may be substituted with an alkoxy group having 1 to 4 carbon atoms, an alkyl group having 1 to 4 carbon atoms, or a halogen atom; R ２ and R ３ may be the same or different and are a hydrogen atom or —(C═O)—R ｃ (In the formula, R ｃ represents a hydroxyl group, an alkoxy group having 1 to 4 carbon atoms, -N(-R １１ ) (-R １２ ) (wherein, R １１ and R １２ each represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms) or —O—(CH ２ )n-(C=O)-R １３ (wherein n is an integer of 1 to 3, and R １３ represents an alkyl group having 1 to 4 carbon atoms), where R ２ and R ３ does not simultaneously represent a hydrogen atom, R ４ and R ７ represents a hydrogen atom, R ５ represents a hydrogen atom or —(C═O)—R ｄ (In the formula, R ｄ represents a hydroxyl group, an alkoxy group having 1 to 4 carbon atoms, -N(-R ２１ ) (-R ２２ ) (wherein, R ２１ and R ２２ each represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms) or —O—(CH ２ )p-(C=O)-R ２３ (wherein p is an integer of 1 to 3, and R ２３ represents an alkyl group having 1 to 4 carbon atoms, R ６ represents a hydrogen atom or —(C═O)—R ｅ (In the formula, R ｅ represents a hydroxyl group, an alkoxy group having 1 to 4 carbon atoms, -N(-R ３１ ) (-R ３２ ) (wherein, R ３１ and R ３２ each represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms) or —O—(CH ２ )r-(C=O)-R ３３ (wherein r is an integer of 1 to 3, R ３３ represents an alkyl group having 1 to 4 carbon atoms.

2. R １ is an alkyl group having 1 to 4 carbon atoms or -R ａ -R ｂ (In the formula, R ａ Ha-CH ２ - or -C(=O)-, R ｂ 2. The compound according to claim 1, or a pharmaceutically acceptable salt or solvate thereof, wherein R represents an optionally substituted aromatic ring group or an optionally substituted aliphatic carbocyclic group.

3. A compound according to claim 1 or 2, or a pharmaceutically acceptable salt or solvate thereof, wherein --- represents a double bond.

4. R ２ and R ３ may be the same or different and represent an alkoxycarbonyl group having 1 to 4 carbon atoms, or a pharmaceutically acceptable salt or solvate thereof.

5. X represents a nitrogen atom, Z represents a carbon atom, R ５ does not exist and R ６ The compound according to claim 1 or 2, or a pharmaceutically acceptable salt or solvate thereof, wherein represents an alkoxycarbonyl group having 1 to 4 carbon atoms.

6. 3. An agent for promoting neutral fat breakdown in adipocytes, comprising the compound of formula (I) according to claim 1 or 2, or a pharmaceutically acceptable salt or solvate thereof, as an active ingredient.

7. A pharmaceutical composition comprising the compound of formula (I) according to claim 1 or 2, or a pharmaceutically acceptable salt or solvate thereof, as an active ingredient.

8. The pharmaceutical composition according to claim 7, for use in treating, preventing or ameliorating a disease or symptom that can be treated, prevented or ameliorated by promoting neutral fat breakdown in adipocytes.

9. The pharmaceutical composition according to claim 8, wherein the disease or condition is obesity or a disease or condition associated therewith.

10. 10. The pharmaceutical composition according to claim 9, wherein the disease and symptom associated with obesity is one or more selected from the group consisting of impaired glucose tolerance, dyslipidemia, hypertension, hyperuricemia, gout, coronary artery disease, cerebral infarction, fatty liver, menstrual disorders, pregnancy complications, sleep apnea syndrome, obesity-hypoventilation syndrome, orthopedic diseases, and obesity-related kidney disease.

11. The pharmaceutical composition according to claim 7 , which is used in combination with an agent for promoting cAMP production in adipocytes.

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