Melanocortin-4 receptor agonist

Abstract

The present invention relates to a compound exhibiting excellent agonistic activity against melanocortin receptors, more specifically, to a compound of formula (1), a pharmaceutical composition containing the compound as an active ingredient, and use thereof. In particular, the compound of the present invention exhibits excellent agonistic activity against the melanocortin-4 receptor, and therefore can be advantageously used in the prevention or treatment of obesity, diabetes, inflammation, and erectile dysfunction.

Description

Technical Field

[0001] The present invention relates to a compound showing excellent agonist activity against melanocortin receptors. More specifically, the present invention relates to a compound represented by the following formula (1):

Chemical Formula

Background Art

[0002] Leptin protein is a hormone secreted by body fat cells (adipocytes), and its secretion amount increases as the body fat rate increases. Leptin protein regulates various in-vivo functions such as appetite, body fat mass and energy metabolism by regulating the functions of various neuropeptides produced in the hypothalamus (Non-Patent Document 1). The signal transduction of appetite and body weight control by leptin protein is carried out through many downstream factors after birth, and the most typical factors are melanocortin, AgRP (agouti-related protein) and neuropeptide Y (NPY) hormones.

[0003] When the concentration of leptin in the blood increases due to excessive calorie intake in the body, the secretion of the proopiomelanocortin (POMC) protein hormone in the pituitary gland increases, and the production of AgRP and NPY decreases. The small peptide hormone α-MSH (melanocyte-stimulating hormone) is produced from POMC neurons, and this hormone is an agonist of the melanocortin-4 receptor (MC4R) of secondary neurons, ultimately inducing anorexia. On the other hand, when the concentration of leptin decreases due to calorie deficiency, the expression of AgRP, which is an antagonist of MC4R, increases, and the expression of NPY also increases, ultimately promoting appetite. That is, depending on the change in leptin, the α-MSH hormone and the AgRP hormone are involved in appetite regulation as an agonist and an antagonist of MC4R.

[0004] In addition to MC4R, the α-MSH hormone induces various physiological reactions by binding to three MCR subtypes. To date, five MCR subtypes have been identified. Among the subtypes, MC1R is mainly expressed in skin cells and is involved in melanin pigmentation, MC2R is mainly expressed in the adrenal glands and is involved in the production of glucocorticoid hormones, and it is known that only ACTH (adrenocorticotropic hormone) derived from POMC is its ligand. MC3R and MC4R, which are mainly expressed in the central nervous system, are involved in the regulation of appetite, energy metabolism, and the efficiency of body fat accumulation, and MC5R, which is expressed in various tissues, is known to regulate exocrine functions (Non-Patent Document 2). Specifically, the activation of the MC4R receptor has been proven to effectively reduce body weight by inducing a decrease in appetite and an increase in energy metabolism, and it is the main target in the development of anti-obesity drugs (Non-Patent Documents 2, 3, 4, 5).

[0005] The role of MC4R in appetite and weight control has been mainly demonstrated through experiments in animal models with abnormal expression of agouti protein (agouti mice). In the case of agouti mice, it has been revealed that due to gene mutation, agouti protein is highly expressed in the central nervous system, acts as an agonist of MC4R in the hypothalamus, and induces obesity (Non-Patent Documents 6 and 7). As a result of subsequent studies, it has been observed that AgRP (agouti-related protein), which is similar to actual agouti protein, is expressed in hypothalamic neurons, and it is also known that AgRP is involved in appetite regulation as an antagonist to MC4R (Non-Patent Documents 8 and 9).

[0006] When α-MSH, an MC4R agonist in vivo, is administered into the brain of animals, it has the effect of reducing appetite. When the MC4R antagonist SHU9119 (peptide) or HS014 (peptide) is administered, the effect of increasing appetite again is observed (Non-Patent Document 10). Also, in animal tests using melanotan II (MTII, Ac-Nle-c[Asp-His-DPhe-Arg-Trp-Lys]-NH2) and an agonist similar to it, HP228, effects such as appetite suppression, weight loss, and increased energy metabolism were confirmed by intracerebral administration, intraperitoneal administration, or subcutaneous administration (Non-Patent Documents 11, 12, and 13). On the other hand, when the typical SHU9119 is administered to animals, significant and sustained feed intake and weight gain are shown, and pharmacological evidence that MCR agonists can be used for the treatment of obesity is obtained. The appetite-reducing effect that is clearly observed during MTII administration is not shown in MC4RKO (knockout) mice, and this experimental result proves again that the appetite-reducing effect is mainly achieved by the activation of MC4R (Non-Patent Document 14).

[0007] As obesity therapeutics developed to date, appetite suppressants that act on the central nervous system are the mainstream, and many of them are drugs that regulate the action of neurotransmitters. For example, noradrenergic agents (phentermine and mazindol), and serotonin-acting agents such as fluoxetine and sibutramine can be mentioned. However, in addition to suppressing appetite by acting on multiple subtypes of receptors, the said neurotransmitter modulators have a wide range of effects on various physiological functions. Therefore, the said modulators have a major drawback in that they lack selectivity for each obese individual and are accompanied by various side effects when administered for a long period of time.

[0008] On the other hand, melanocortin is a neuropeptide rather than a neurotransmitter, and considering that MC4R gene KO mice have normal functions other than energy metabolism, melanocortin agonists have the advantage as a point of action that they can induce only weight loss by suppressing appetite without affecting other physiological functions. In particular, its receptor is a G-protein coupled receptor (GPCR), and it is significantly different from the points of action of the prior art in that selectivity for subtype receptors can be relatively easily ensured.

[0009] As an example of utilizing such a melanocortin receptor as a point of action, Patent Document 1 and Patent Document 2 disclose compounds as agonists of the melanocortin receptor.

Prior Art Documents

Patent Documents

[0010]

Patent Document 1

Patent Document 2

Non-Patent Documents

[0011]

Non-Patent Document 1

Non-Patent Document 14

Summary of the Invention

Problems to be Solved by the Invention

[0012] An object of the present invention is to provide a novel compound represented by formula (1) having excellent selective agonist activity against melanocortin receptors, particularly melanocortin-4 receptor (MC4R), or a pharmaceutically acceptable salt or isomer thereof.

[0013] Another object of the present invention is to provide a method for producing the compound represented by formula (1).

[0014] Yet another object of the present invention is to provide a melanocortin receptor agonist pharmaceutical composition containing, as an active ingredient, the compound represented by formula (1), or a pharmaceutically acceptable salt or isomer thereof.

[0015] Still another object of the present invention is to provide the use of the compound represented by formula (1), or a pharmaceutically acceptable salt or isomer thereof, in the prevention or treatment of obesity, diabetes, inflammation and erectile dysfunction.

Means for Solving the Problems

[0016] To achieve the above object, in the present invention, the following formula (1)

Chemical Formula

[0017] The compound of formula (1) according to the present invention can form pharmaceutically acceptable salts.

[0018] In addition, since the compounds according to the present invention may have asymmetric carbon centers and asymmetric axes or planes, they may exist as cis or trans isomers, R or S isomers, racemates, mixtures of diastereomers, and individual diastereomers, and all of these isomers and mixtures are included within the scope of the present invention.

[0019] In this specification, unless otherwise indicated, the compound of formula (1) is used in the sense of including all of the compound of formula (1), its pharmaceutically acceptable salts and isomers.

[0020] As used herein, the term "halo" or "halogen" means a radical of fluorine (F), chlorine (Cl), bromine (Br) or iodine (I).

[0021] As used herein, the term "alkyl" means a straight-chain or branched hydrocarbon group.

[0022] In one embodiment according to the present invention, R1 of the formula (1) is C2-C4 alkyl. In another embodiment according to the present invention, R1 of the formula (1) is straight-chain or branched C2-C4 alkyl, for example, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl.

[0023] In another embodiment according to the present invention, R1 of the formula (1) is C3 or C4 alkyl. In another embodiment according to the present invention, R1 of the formula (1) is branched C3 or C4 alkyl, for example, isopropyl or tert-butyl.

[0024] In another embodiment according to the present invention, the compound of the formula (1) is the following formula (2)

Chemical formula

[0025] In another embodiment of the present invention, the compound of formula (1) is the following formula (3)

Chemical formula

[0026] In another embodiment of the present invention, the compound of formula (1) is the following formula (4)

Chemical formula

[0027] In another embodiment of the present invention, the compound of formula (1) is the following formula (5)

Chemical formula

[0028] In another embodiment of the present invention, the compound of formula (1) is the following formula (6) [Chemical formula] N-((3S,5S)-1-((3S,4R)-1-(tert-Butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-(4,4-dimethylcyclohexyl)isobutyramide represented by

[0029] In another embodiment of the present invention, the compound of formula (1) is the following formula (7) [Chemical formula] N-((3S,5S)-1-((3S,4R)-1-(tert-Butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-(4,4-dimethylcyclohexyl)pivalamide represented by

[0030] In another embodiment according to the present invention, the pharmaceutically acceptable salts include, but are not limited to, acid addition salts formed by inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrobromic acid, hydroiodic acid, organic carboxylic acids such as tartaric acid, formic acid, citric acid, acetic acid, trichloroacetic acid, trifluoroacetic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, maleic acid, and sulfonic acids such as methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or naphthalenesulfonic acid.

[0031] In another embodiment according to the present invention, the pharmaceutically acceptable salt is a hydrochloride.

[0032] In another embodiment of the present invention, the compound of formula (1) is the following formula (8) [Chemical formula] It is N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(2,4-difluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)pivalamide hydrochloride.

[0033] In another embodiment according to the present invention, the compound of formula (1) is the following formula (9)

Chemical formula

[0034] In another embodiment according to the present invention, the compound of formula (1) is the following formula (10)

Chemical formula

[0035] In another embodiment according to the present invention, the compound of formula (1) is the following formula (11)

Chemical formula

[0036] In another embodiment according to the present invention, the compound of formula (1) is the following formula (12)

Chemical formula

[0037] In another embodiment according to the present invention, the compound of formula (1) is the following formula (13)

Chemical formula

[0038] In another embodiment according to the present invention, the N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(2,4-difluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)pivalamide hydrochloride of formula (8),

[0039] the N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(2,4-difluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)isobutylamide hydrochloride of formula (9),

[0040] N-((3S,5S)-1-((3S,4R)-1-(tert-Butyl)-4-(4-chloro-2-fluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)isobutylamide hydrochloride of the formula (1) 0 and

[0041] The N-((3S,5S)-1-((3S,4R)-1-(tert-Butyl)-4-(4-chloro-2-fluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)pivalamide hydrochloride of the formula (11) can be produced according to the following reaction scheme 1. <Reaction Scheme 1> [Chemical formula] (In the formula, R1, R2 and R3 have the same meanings as defined above.) In another embodiment according to the present invention, the N-((3S,5S)-1-((3S,4R)-1-(tert-Butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-(4,4-dimethylcyclohexyl)isobutylamide hydrochloride of the formula (12) and

[0042] The N-((3S,5S)-1-((3S,4R)-1-(tert-Butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-(4,4-dimethylcyclohexyl)pivalamide hydrochloride of the formula (13) can be produced according to the following reaction scheme 2. <Reaction Scheme 2> [Chemical formula] (R1 has the same meaning as defined above.)

[0043] The compound of formula (1) according to the present invention exhibits excellent agonist activity against melanocortin receptors, particularly melanocortin-4 receptor (MC4R). Therefore, the present invention also provides an agonist pharmaceutical composition for melanocortin receptors, which comprises, as an active ingredient, the compound of formula (1), or a pharmaceutically acceptable salt or isomer thereof, together with a pharmaceutically acceptable carrier. In particular, the composition according to the present invention exhibits excellent effects in the prevention or treatment of obesity, diabetes, inflammation and erectile dysfunction, but is not limited thereto.

[0044] As used herein, the term "carrier" means a compound that facilitates the injection of a compound into cells or tissues.

[0045] When the compound of the present invention is administered for clinical purposes, the total daily dose administered to the host in a single or divided dose is preferably in the range of 0.01 to 10 mg per kg of body weight. However, the specific dose for an individual patient varies depending on the specific compound used, the patient's body weight, gender, health status, diet, administration time of the drug, administration method, excretion rate, drug combination and severity of the disease, etc.

[0046] The compound of the present invention can be administered by any route according to the purpose. For example, the compound of the present invention can be administered by injection or oral administration.

[0047] Injectable preparations can be manufactured according to known techniques by using appropriate dispersants, wetting agents or suspending agents.

[0048] Examples of solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. The solid dosage forms can be manufactured by mixing the active compound of formula (1) according to the present invention with one or more carriers such as an inert diluent, lubricant, disintegrant, binder, etc.

Advantages of the Invention

[0049] The compound of formula (1) according to the present invention exhibits excellent agonist activity against melanocortin receptors, particularly melanocortin-4 receptor (MC4R), and thus can be conveniently and usefully used for the prevention or treatment of obesity, diabetes, inflammation and erectile dysfunction.

[0050] The compound of formula (1) according to the present invention shows an on-target effect on the melanocortin-4 receptor, exhibits weight loss and diet effects, does not affect anxiety and depression, and can be administered without safety problems such as side effects of hERG inhibition and mutagenesis. In addition, the compound of formula (1) according to the present invention can be safely administered because it has no cytotoxicity and hepatotoxicity.

BEST MODE FOR CARRYING OUT THE INVENTION

[0051] Hereinafter, the present invention will be described in more detail with reference to the following examples. However, it should be understood that the protection scope of the present invention is not limited to these examples.

[0052] Production Example 1: Production of N-((1s,4R)-4-methylcyclohexyl)-N-((3S,5S)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)pivalamide hydrochloride

Chemical formula

[0053] The title compound was obtained through the following steps A, B and C. Step A: Production of (2S,4S)-1-(tert-butoxycarbonyl)-4-(N-((1s,4R)-4-methylcyclohexyl)pivalamide)pyrrolidine-2-carboxylic acid

Chemical formula

[0054] The title compound was obtained by the method disclosed in International Publication No. WO 2008 / 007930. MS [M+Na] = 433.4 (M+23) 1 H NMR (400 MHz, CD3OD) δ 4.25 (m, 1H), 3.86 (m, 2H), 3.42 (m, 2H), 2.80 (m, 1H), 2.27 (m, 1H), 2.00 - 1.80 (m, 3H), 1.66 (m, 4H), 1.43 (m, 11H), 1.26 (m, 9H), 1.05 (d, 3H)

[0055] Process B: Preparation of tert - butyl (2S,4S) - 4 - (N - ((1s,4R) - 4 - methylcyclohexyl)pivalamido) - 2 - (morpholine - 4 - carbonyl)pyrrolidine - 1 - carboxylate

Chemical formula

[0056] (2S,4S) - 1 - (tert - butoxycarbonyl) - 4 - (N - ((1s,4R) - 4 - methylcyclohexyl)pivalamido)pyrrolidine - 2 - carboxylic acid (0.81 g, 1.97 mmol) obtained in the above Process A was dissolved in 10 mL of dimethylformamide, and morpholine (0.19 mL, 2.17 mmol), 1H - benzotriazol - 1 - ol monohydrate (0.36 g, 2.36 mmol), hydrochloride of 3 - (((ethylimino)methylene)amino) - N,N - dimethylpropan - 1 - amine (0.45 g, 2.36 mmol) and N,N - diisopropylethylamine (1.0 mL, 5.92 mmol) were added, and the mixture was stirred at room temperature for 16 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure, the filtrate was washed with an aqueous sodium hydrogen carbonate solution, and extracted with ethyl acetate. The organic layer was collected, washed with water, dried over anhydrous magnesium sulfate, and the solid was filtered off. The filtrate was concentrated under reduced pressure and purified by column chromatography to obtain the title compound (0.62 g, 66%). MS [M+H] = 480.5 (M+1) 11H NMR (400 MHz, CD3OD) δ 4.65 (m, 1H), 4.00 - 3.40 (m, 12H), 2.82 (m, 1H), 2.22 (m, 1H), 2.00 - 1.80 (m, 3H), 1.70 - 1.60 (m, 4H), 1.50 - 1.41 (m, 11H), 1.24 (s, 9H), 1.04 (d, 3H)

[0057] Process C: Preparation of N - ((1S,4R) - 4 - methylcyclohexyl) - N - ((3S,5S) - 5 - (morpholine - 4 - carbonyl)pyrrolidin - 3 - yl)pivalamide hydrochloride

Chemical Structure

[0058] tert - Butyl (2S,4S) - 4 - (N - ((1S,4R) - 4 - methylcyclohexyl)pivalamide) - 2 - (morpholine - 4 - carbonyl)pyrrolidine - 1 - carboxylate (0.62 g, 1.3 mmol) obtained in the above Process B was dissolved in 4 mL of dichloromethane, and a 1,4 - dioxane solution of 4M hydrochloric acid (1.3 mL, 5.1 mmol) was added. The mixture was stirred at room temperature for 16 hours. The reaction solution was concentrated under reduced pressure, and the resulting solid was washed with diethyl ether and dried to obtain the title compound (0.55 g, 99%). MS [M + H] = 380.6 (M + 1) 1 1H NMR (400 MHz, DMSO - d6) δ 10.00 - 8.00 (brs, 2H), 4.46 (m, 1H), 4.14 (m, 1H), 3.76 (m, 1H), 3.65 - 3.25 (m, 10H), 2.47 (m, 1H), 1.99 (m, 1H), 1.91 (m, 1H), 1.80 - 1.50 (m, 6H), 1.39 (m, 2H), 1.19 (s, 9H), 1.00 (d, 3H)

[0059] Production Example 2: Production of N-((1S,4R)-4-methylcyclohexyl)-N-((3S,5S)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)isobutylamide hydrochloride

Chem.

[0060] The title compound was obtained through the following steps A, B, C, D, E, F, and G. Step A: Production of 1-(tert-butyl) 2-methyl (2S,4S)-4-azidopyrrolidine-1,2-dicarboxylate

Chem.

[0061] 1-(tert-Butyl) 2-methyl (2S,4R)-4-((methylsulfonyl)oxy)pyrrolidine-1,2-dicarboxylate (48.5 g, 150 mmol) was dissolved in 250 mL of dimethylformamide, and sodium azide (19.5 g, 300 mmol) was added. After stirring the mixture at 80 °C for 16 hours, the solution was concentrated under reduced pressure. Water was added thereto, and the mixture was extracted with ethyl acetate. The organic layer was washed with an aqueous sodium chloride solution and water, dried over anhydrous magnesium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain the title compound (39.59 g, 98%), which was used in the next step without purification. MS [M+H] = 271 (M+1) 1 H NMR (400 MHz, CD3OD) δ 4.43-4.37 (m, 1H), 4.35-4.27 (br, 1H), 3.77 (s, 1.8H), 3.76 (s, 1.2H), 3.73-3.66 (m, 1H), 3.44-3.38 (m, 1H), 2.63-2.49 (m, 1H), 2.19-2.11 (m, 1H), 1.50 (s, 4.5H), 1.44 (s, 4.5H)

[0062] Process B: Production of 1-(tert-butyl) 2-methyl (2S,4S)-4-aminopyrrolidine-1,2-dicarboxylate

Chem.

[0063] 1-(tert-butyl) 2-methyl (2S,4S)-4-azidopyrrolidine-1,2-dicarboxylate (24.6 g, 91.0 mmol) obtained in the above Process A was dissolved in 180 mL of tetrahydrofuran, and 1 M trimethylphosphine tetrahydrofuran solution (109 mL, 109 mmol) was slowly added thereto at 0 °C. After stirring at the same temperature for 1 hour, the reaction mixture was stirred at room temperature for 3 hours. After concentrating the reaction solvent under reduced pressure, 100 mL of dichloromethane and 150 mL of water were added, and the reaction mixture was stirred for about 30 minutes. The organic layer was dried over anhydrous magnesium sulfate and the solid was filtered off. The filtrate was concentrated under reduced pressure to obtain the title compound (20.62 g, 93%). MS [M+H] = 245 (M+1) 1 H NMR (400 MHz, CD3OD) δ 4.27 (m, 1H), 3.77 (s, 1.8H), 3.76 (s, 1.2H), 3.75 - 3.67 (m, 1H), 3.50 - 3.42 (m, 1H), 3.22 - 3.17 (m, 1H), 2.58 - 2.47 (m, 1H), 1.82 - 1.71 (m, 1H), 1.48 (s, 4.5H), 1.42 (s, 4.5H)

[0064] Process C: Production of 1-(tert-butyl) 2-methyl (2S,4S)-4-(((1s,4R)-4-methylcyclohexyl)amino)pyrrolidine-1,2-dicarboxylate

Chem.

[0065] 1-(tert-Butyl) 2-methyl (2S,4S)-4-aminopyrrolidine-1,2-dicarboxylate (20.6 g, 84.4 mmol) obtained in the above step B was dissolved in 150 mL of 1,2-dichloroethane, and 4-methylcyclohexanone (9.50 mL, 101 mmol) was added. After cooling the mixture to 0 °C, sodium triacetoxyborohydride (26.8 g, 127 mmol) was added, and the mixture was stirred at room temperature for 16 hours. After concentrating the reaction solution under reduced pressure, water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with an aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and filtered. The filtrate was concentrated under reduced pressure and purified by column chromatography to obtain the title compound (22.9 g, 80%). MS [M+H] = 341 (M+1) 1 H NMR (400 MHz, CD3OD) δ 4.26 (m, 1H), 3.76 (s, 1.8H), 3.75 (s, 1.2H), 3.78-3.71 (m, 1H), 3.49-3.40 (m, 1H), 3.22-3.16 (m, 1H), 2.69-2.60 (br, 1H), 2.58-2.46 (m, 1H), 1.87-1.77 (m, 1H), 1.73-1.63 (m, 1H), 1.62-1.35 (m, 8H), 1.48 (s, 4.5H), 1.42 (s, 4.5H), 0.96 (d, 3H)

[0066] Step D: Preparation of 1-(tert-Butyl) 2-methyl (2S,4S)-4-(N-((1s,4R)-4-methylcyclohexyl)isobutylamide)pyrrolidine-1,2-dicarboxylate

Chemical Structure

[0067] 1-(tert-Butyl) 2-methyl (2S,4S)-4-(((1s,4R)-4-methylcyclohexyl)amino)pyrrolidine-1,2-dicarboxylate (37.29 g, 109.5 mmol) obtained in the above step C was dissolved in 500 mL of dichloromethane. After adding triethylamine (61.1 mL, 438 mmol), isobutyl chloride (11.7 mL, 219 mmol) was slowly added at 0 °C. After stirring at room temperature for 16 hours, the reaction solvent was concentrated under reduced pressure. An aqueous sodium bicarbonate solution and ethyl acetate were added to the concentrated solution, and then the organic layer was separated. The organic layer was washed with an aqueous sodium chloride solution and water, dried over anhydrous magnesium sulfate, and filtered. The filtrate was concentrated under reduced pressure and purified by column chromatography to obtain the title compound (38.79 g, 86%). MS [M+H] = 411 (M+1) 1 H NMR (400 MHz, CD3OD) δ 4.27 (m, 1H), 3.76 (s, 1.8H), 3.75 (s,1.2H), 3.78 - 3.72 (m, 1H), 3.50 - 3.41 (m, 1H), 3.33 - 3.14 (m, 1H), 2.69 - 2.60 (m, 2H), 2.57 - 2.43 (m, 1H), 1.87 - 1.79 (m, 1H), 1.70 - 1.61 (m, 1H), 1.60 - 1.32 (m, 8H), 1.47 (s, 4.5H), 1.41 (s, 4.5H), 1.10 (dd, 6H), 0.99 (d, 3H)

[0068] Step E: Preparation of (2S,4S)-1-(tert-butoxycarbonyl)-4-(N-((1s,4R)-4-methylcyclohexyl)isobutylamide)pyrrolidine-2-carboxylic acid

Chemical Structure

[0069] 1-(tert-Butyl) 2-methyl (2S,4S)-4-(N-((1s,4R)-4-methylcyclohexyl)isobutylamide)pyrrolidine-1,2-dicarboxylate (3.63 g, 8.85 mmol) obtained in the above step D was dissolved in 30 mL of ethanol, 1N aqueous sodium hydroxide solution (26.5 mL, 26.5 mmol) was added, and the mixture was stirred at room temperature for 2 hours. After diluting the reaction solution with water, it was adjusted to pH 4 with 1N aqueous hydrochloric acid solution. The reaction solution was extracted with ethyl acetate, the organic layer was separated, and dried over anhydrous sodium sulfate. The solid was filtered, and the filtrate was concentrated under reduced pressure to obtain the title compound (2.40 g, 69%). MS [M+Na] = 419.4 (M+23) 1 H NMR (400 MHz, CD3OD) δ 4.27 (m, 1H), 4.00 - 3.85 (m, 2H), 3.66 (m, 1H), 3.44 (m, 1H), 2.86 (m, 1H), 1.95 - 1.80 (m, 3H), 1.76 - 1.53 (m, 5H), 1.50 - 1.42 (m, 11H), 1.10 - 1.05 (m, 9H)

[0070] Step F: Preparation of tert-butyl (2S,4S)-4-(N-((1s,4R)-4-methylcyclohexyl)isobutylamide)-2-(morpholine-4-carbonyl)pyrrolidine-1-carboxylate

Chemical Structure

[0071] (2S,4S)-1-(tert-Butoxycarbonyl)-4-(N-((1s,4R)-4-methylcyclohexyl)isobutylamide)pyrrolidine-2-carboxylic acid (2.40 g, 6.05 mmol), 1H-benzo[d][1,2,3]triazol-1-ol monohydrate (1.11 g, 7.26 mmol), and hydrochloride of 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine (1.39 g, 7.26 mmol) were dissolved in 30 mL of dimethylformamide. Morpholine (0.55 mL, 6.66 mmol) and N,N-diisopropylethylamine (3.10 mL, 18.2 mmol) were added to the above mixture, and the mixture was stirred at room temperature for 16 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure, the filtrate was washed with an aqueous sodium hydrogen carbonate solution, and extracted with ethyl acetate. The organic layer was collected, washed with water, dried over anhydrous magnesium sulfate, and the solid was filtered. The filtrate was concentrated under reduced pressure and purified by column chromatography to obtain the title compound (1.87 g, 66%). MS [M+H] = 466.5 (M+1) 1 H NMR (400 MHz, CD3OD) δ 4.66 (m, 1H), 3.94 (m, 1H), 3.75 - 3.55 (m, 10H), 3.45 (m, 1H), 2.86 (m, 1H), 2.24 (m, 1H), 1.95 - 1.80 (m, 3H), 1.77 - 1.60 (m, 5H), 1.59 - 1.45 (m, 2H), 1.46 - 1.41 (m, 9H), 1.05 (m, 9H)

[0072] Step G: Preparation of N-((1s,4R)-4-methylcyclohexyl)-N-((3S,5S)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)isobutylamide hydrochloride

Chemical Structure

[0073] The tert-butyl (2S,4S)-4-(N-((1s,4R)-4-methylcyclohexyl)isobutylamide)-2-(morpholine-4-carbonyl)pyrrolidine-1-carboxylate (1.87 g, 4.00 mmol) obtained in the above step F was dissolved in 30 mL of dichloromethane, cooled to 0 °C, and 4M hydrochloric acid 1,4-dioxane (2.15 mL, 8.59 mmol) was added. After the mixture was stirred at room temperature for 16 hours, the reaction solution was concentrated under reduced pressure. The resulting solid was washed with ethyl ether and dried to obtain the title compound (1.22 g, 76%). MS [M+H] = 366.4 (M+1) 1 H NMR (400 MHz, DMSO-d6) δ 9.88 (brs, 1H), 8.12 (m, 1H), 4.51 (m, 1H), 4.20 (m, 1H), 3.63-3.35 (m, 10H), 3.30 (m, 1H), 2.85 (m, 1H), 2.51 (m, 1H), 2.01 (m, 1H), 1.91 (m, 1H), 1.76-1.60 (m, 4H), 1.53 (m, 2H), 1.40 (m, 2H), 0.99 (m, 9H)

[0074] Production Example 3: Preparation of methyl (2S,4S)-4-(N-(4,4-dimethylcyclohexyl)pivalamide)pyrrolidine-2-carboxylate hydrochloride

Chemical formula

[0075] Step A: Preparation of 1-(tert-butyl) 2-methyl (2S,4S)-4-(N-(4,4-dimethylcyclohexyl)pivalamide)pyrrolidine-1,2-dicarboxylate

Chemical formula

[0076] The title compound was obtained by the method disclosed in International Publication No. WO 2008 / 007930. MS [M+Na] = 461.4 (M+23) 1 H NMR (400 MHz, CD3OD) δ 4.34 (t, 1H), 3.90 - 3.70 (m, 2H), 3.73 (m, 3H), 3.45 (m, 2H), 2.74 - 2.61 (m, 1H), 2.30 (m, 1H), 1.83 (m, 2H), 1.53 (m, 4H), 1.45 - 1.40 (m, 9H), 1.40 - 1.30 (m, 2H), 1.24 (s, 9H), 0.99 (s, 3H), 0.94 (s, 3H)

[0077] Step B: Preparation of methyl (2S,4S)-4-(N-(4,4-dimethylcyclohexyl)pivalamide)pyrrolidine-2-carboxylate hydrochloride

Chemical Structure

[0078] 1-(tert-Butyl) 2-methyl (2S,4S)-4-(N-(4,4-dimethylcyclohexyl)pivalamide)pyrrolidine-1,2-dicarboxylate (1.67 g, 3.81 mmol) obtained in Step A above was dissolved in 4 mL of ethyl acetate, and 4M hydrochloric acid ethyl acetate solution (3.81 mL, 15.2 mmol) was added. The mixture was stirred at room temperature for 16 hours. The reaction solution was concentrated under reduced pressure to obtain the title compound (1.43 g, 99%). MS [M+H] = 339.4 (M+1) 1 H NMR (400 MHz, CD3OD) δ 4.41 (t, 1H), 4.24 (m, 1H), 3.86 (m, 1H), 3.83 (s, 3H), 3.42 (m, 2H), 2.66 (m, 1H), 2.22 (m, 1H), 1.77 (m, 2H), 1.51 (m, 4H), 1.34 (m, 2H), 1.23 (s, 9H), 0.99 (s, 3H), 0.93 (s, 3H)

[0079] Production Example 4: Production of Methyl (2S,4S)-4-(N-(4,4-dimethylcyclohexyl)isobutylamide)pyrrolidine-2-carboxylate Hydrochloride

Chemical formula

[0080] Step A: Production of 1-(tert-butyl) 2-methyl (2S,4S)-4-(N-(4,4-dimethylcyclohexyl)isobutylamide)pyrrolidine-1,2-dicarboxylate

Chemical formula

[0081] The title compound was obtained by the method disclosed in International Publication No. WO 2008 / 007930. MS [M+Na] = 447.4 (M+23) 1 H NMR (400 MHz, CD3OD) δ 4.31 (t, 1H), 3.93 (m, 1H), 3.83 (m, 1H), 3.75 - 3.68 (m, 3H), 3.62 (m, 1H), 3.45 (m, 1H), 2.81 (m, 1H), 2.26 (m, 1H), 1.80 (m, 2H), 1.50 - 1.35 (m, 14H), 1.21 (m, 2H), 1.04 (m, 6H), 0.96 (s, 3H), 0.92 (s, 3H)

[0082] Step B: Production of Methyl (2S,4S)-4-(N-(4,4-dimethylcyclohexyl)isobutylamide)pyrrolidine-2-carboxylate Hydrochloride

Chemical formula

[0083] 1-(tert-Butyl) 2-methyl (2S,4S)-4-(N-(4,4-dimethylcyclohexyl)isobutylamide) pyrrolidine-1,2-dicarboxylate (1.1 g, 2.6 mmol) obtained in the above step A was dissolved in 2.5 mL of ethyl acetate, and 4M hydrochloric acid ethyl acetate solution (2.5 mL, 10 mmol) was added. After stirring the reaction solution at room temperature for 16 hours, it was concentrated under reduced pressure to obtain the title compound (0.93 g, 99%). MS [M+H] = 325.4 (M+1) 1 H NMR (400 MHz, CD3OD) δ 4.41 (t, 1H), 4.30 (m, 1H), 3.84 (s, 3H), 3.66 (m, 1H), 3.47 (m, 3H), 2.85 (m, 1H), 2.72 (m, 1H), 2.24 (m, 1H), 1.75 (m, 2H), 1.51 (m, 4H), 1.38 (m, 2H), 1.06 (m, 6H), 0.98 (s, 3H), 0.93 (s, 3H)

[0084] Production Example 5: Preparation of (3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carboxylic acid

Chemical formula

[0085] The title compound was obtained by the method disclosed in International Publication No. WO 2004 / 092126 MS [M+H] = 282 (M+1) 1 H NMR (400 MHz, CD3OD) δ 7.43-7.33 (m, 4H), 3.90-3.69 (m, 3H), 3.59 (m, 1H), 3.29 (m, 1H), 3.18-3.09 (m, 1H), 1.44 (s, 9H)

[0086] Production Example 6: Preparation of (3S,4R)-1-(tert-butyl)-4-(2,4-difluorophenyl)pyrrolidine-3-carboxylic acid

Chem.

[0087] The title compound was obtained by the method disclosed in International Publication No. WO 2004 / 092126. MS [M+H] = 284.2 (M+1) 1 H NMR (400 MHz, CD3OD) δ 7.50 (m, 1H), 6.97 (m, 2H), 3.93 - 3.75 (m, 3H), 3.60 (m, 1H), 3.26 (m, 2H), 1.43 (s, 9H)

[0088] Production Example 7: Production of (3S,4R)-1-(tert-butyl)-4-(4-chloro-2-fluorophenyl)pyrrolidine-3-carboxylic acid

Chem.

[0089] The title compound was obtained by the method disclosed in International Publication No. WO 2004 / 092126. MS [M+H] = 300.3 (M+1) 1 H NMR (400 MHz, CD3OD) δ 7.47 (t, 1H), 7.22 (m, 2H), 3.93 - 3.75 (m, 3H), 3.60 (m, 1H), 3.26 (m, 2H), 1.43 (s, 9H)

[0090] Example 1: Production of N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(2,4-difluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)pivalamide hydrochloride

Chem.

[0091] The title compound was obtained through the following processes of step A and step B. Step A: Preparation of N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(2,4-difluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)pivalamide

Chemical formula

[0092] To N-((1s,4R)-4-methylcyclohexyl)-N-((3S,5S)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)pivalamide hydrochloride (0.50 g, 1.2 mmol) obtained in Production Example 1 and (3S,4R)-1-(tert-butyl)-4-(2,4-difluorophenyl)pyrrolidine-3-carboxylic acid (0.34 g, 1.2 mmol) obtained in Production Example 6 were added dimethylformamide (12 mL), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.28 g, 1.5 mmol), 1-hydroxybenzotriazole hydrate (0.22 g, 1.5 mmol) and N,N-diisopropylethylamine (0.64 mL, 3.6 mmol), and the mixture was stirred at room temperature for 16 hours. After concentrating the reaction solution under reduced pressure, ethyl acetate was added. The reaction mixture was washed with an aqueous ammonium chloride solution and a 1M aqueous sodium hydroxide solution, and dried over anhydrous sodium sulfate. The solid was filtered, the filtrate was concentrated under reduced pressure, and purified by chromatography to obtain the title compound (0.32 g, 40%). MS [M+H] = 645.6 (M+1) 11H NMR (400 MHz, CD3OD) δ 7.58 (m, 1H), 6.95 (m, 2H), 4.76 (m, 1H), 4.18 (m, 1H), 3.84 - 3.40 (m, 15H), 3.11 (m, 1H), 2.89 (m, 1H), 2.12 (m, 1H), 1.95 (m, 1H), 1.79 (m, 1H), 1.65 (m, 4H), 1.44 (m, 2H), 1.28 (m, 2H), 1.19 (s, 9H), 1.17 (s, 9H), 1.03 (m, 3H)

[0093] Process B: Preparation of N - ((3S,5S) - 1 - ((3S,4R) - 1 - (tert - butyl) - 4 - (2,4 - difluorophenyl)pyrrolidine - 3 - carbonyl) - 5 - (morpholine - 4 - carbonyl)pyrrolidin - 3 - yl) - N - ((1s,4R) - 4 - methylcyclohexyl)pivalamide hydrochloride

Chemical Structure

[0094] To N - ((3S,5S) - 1 - ((3S,4R) - 1 - (tert - butyl) - 4 - (2,4 - difluorophenyl)pyrrolidine - 3 - carbonyl) - 5 - (morpholine - 4 - carbonyl)pyrrolidin - 3 - yl) - N - ((1s,4R) - 4 - methylcyclohexyl)pivalamide (0.32 g, 0.50 mmol) obtained in the above Process A, 5 mL of dichloromethane and 4M hydrochloric acid ethyl acetate solution (0.25 mL, 1.0 mmol) were added, and the mixture was stirred at room temperature for 20 minutes. The reaction solution was concentrated under reduced pressure, and diethyl ether was added. The obtained solid was filtered and dried to obtain the title compound (0.23 g, 68%). MS [M + H] = 645.6 (M + 1) 11H NMR (400 MHz, CD3OD) δ 7.63 (m, 1H), 7.07 (m, 2H), 4.82 (m, 1H), 4.19 (m, 1H), 4.00 - 3.50 (m, 15H), 3.03 (m, 1H), 2.69 (m, 1H), 2.17 (m, 1H), 1.95 (m, 1H), 1.79 (m, 1H), 1.64 (m, 4H), 1.47 (s, 9H), 1.45 - 1.25 (m, 4H), 1.19 (s, 9H), 1.00 (m, 3H)

[0095] Example 2: Preparation of N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(2,4-difluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)isobutylamide hydrochloride

Chemical Structure

[0096] The title compound was obtained through the following steps A and B. Step A: Preparation of N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(2,4-difluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)isobutylamide

Chemical Structure

[0097] (3S,4R)-1-(tert-Butyl)-4-(2,4-difluorophenyl)pyrrolidine-3-carboxylic acid (0.50 g, 1.76 mmol) obtained in Production Example 6 and N-((1s,4R)-4-methylcyclohexyl)-N-((3S,5S)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)isobutylamide hydrochloride (0.71 g, 1.76 mmol) obtained in Production Example 2 were used, and the title compound (0.10 g, 9%) was obtained in the same manner as in Step A of Example 1. MS [M+H] = 631.5 (M+1) 1 H NMR (400 MHz, CD3OD) δ 7.56 (m, 1H), 7.00 (m, 2H), 4.79 (m, 1H), 4.56 (m, 1H), 4.17 (m, 1H), 3.84 (m, 1H), 3.70 - 3.50 (m, 13H), 3.09 (m, 1H), 2.80 (m, 1H), 2.70 (m, 1H), 2.12 (m, 1H), 1.93 (m, 1H), 1.75 - 1.50 (m, 5H), 1.40 (m, 2H), 1.29 (s, 9H), 1.24 (m, 2H), 1.00 (m, 9H)

[0098] Step B: Production of hydrochloride of N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(2,4-difluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)isobutylamide

Chemical formula

[0099] Using N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(2,4-difluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)isobutylamide (0.10 g, 0.16 mmol) obtained in the above step A, the title compound (0.083 g, 78%) was obtained in the same manner as in step B of Example 1. MS [M+H] = 631.5 (M+1) 1 H NMR (400 MHz, CD3OD) δ 7.63 (m, 1H), 7.07 (m, 2H), 4.80 (t, 1H), 4.21 (m, 1H), 3.94 (m, 2H), 3.85 - 3.50 (m, 13H), 3.03 (m, 1H), 2.82 (m, 1H), 2.70 (m, 1H), 2.15 (m, 1H), 1.95 (m, 1H), 1.80 - 1.59 (m, 5H), 1.47 (s, 9H), 1.40 - 1.20 (m, 4H), 1.01 (m, 9H)

[0100] Example 3: Preparation of N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chloro-2-fluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)isobutylamide hydrochloride

Chemical formula

[0101] The title compound was obtained through the following steps A and B. Step A: Preparation of N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chloro-2-fluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)isobutylamide

Chemical formula

[0102] (3S,4R)-1-(tert-Butyl)-4-(4-chloro-2-fluorophenyl)pyrrolidine-3-carboxylic acid (0.50 g, 1.67 mmol) obtained in Production Example 7 and N-((1s,4R)-4-methylcyclohexyl)-N-((3S,5S)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)isobutylamide hydrochloride (0.67 g, 1.67 mmol) obtained in Production Example 2 were used, and the title compound (0.13 g, 12%) was obtained in the same manner as in Step A of Example 1. MS [M+H] = 647.5 (M+1) 1 H NMR (400 MHz, CD3OD) δ 7.54 (t, 1H), 7.20 (m, 2H), 4.79 (m, 1H), 4.18 (m, 1H), 3.85 (m, 1H), 3.50 - 3.50 (m, 13H), 3.40 (m, 1H), 3.02 (m, 1H), 2.78 (m, 1H), 2.68 (m, 1H), 2.14 (m, 1H), 1.92 (m, 1H), 1.80 - 1.39 (m, 7H), 1.29 - 1.17 (m, 11H), 1.01 (m, 9H)

[0103] Step B: Production of N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chloro-2-fluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)isobutylamide hydrochloride

Chemical formula

[0104] Example 4: Preparation of N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chloro-2-fluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)pivalamide hydrochloride

Chemical formula

[0105] The title compound was obtained through the following Steps A and B. Step A: Preparation of N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chloro-2-fluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)pivalamide

Chemical formula

[0106] (3S,4R)-1-(tert-Butyl)-4-(4-chloro-2-fluorophenyl)pyrrolidine-3-carboxylic acid (0.36 g, 1.21 mmol) obtained in Production Example 7 and N-((1s,4R)-4-methylcyclohexyl)-N-((3S,5S)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)pivalamide hydrochloride (0.50 g, 1.21 mmol) obtained in Production Example 1 were used, and the title compound (0.45 g, 56%) was obtained in the same manner as in Step A of Example 1. MS [M+H] = 661.6 (M+1) 1 H NMR (400 MHz, CD3OD) δ 7.55 (m, 1H), 7.18 (m, 2H), 4.79 (m, 1H), 4.18 (m, 1H), 3.90 - 3.20 (m, 15H), 3.08 (m, 1H), 2.70 (m, 1H), 2.12 (m, 1H), 1.93 (m, 1H), 1.81 - 1.60 (m, 5H), 1.47 (m, 2H), 1.30 - 1.10 (m, 2H), 1.20 (s, 9H), 1.15 (s, 9H), 1.03 (m, 3H)

[0107] Step B: Production of N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chloro-2-fluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)pivalamide hydrochloride

Chemical formula

[0108] Using N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chloro-2-fluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)pivalamide (0.25 g, 0.38 mmol), the title compound (0.18 g, 68%) was obtained in the same manner as in Step B of Example 1. MS [M+H] = 661.6 (M+1) 1 H NMR (400 MHz, CD3OD) δ 7.60 (t, 1H), 7.30 (m, 2H), 4.80 (m, 1H), 4.20 (m, 1H), 4.04 - 3.90 (m, 2H), 3.86 - 3.40 (m, 13H), 3.12 (m, 1H), 2.67 (m, 1H), 2.16 (m, 1H), 1.94 (m, 1H), 1.82 - 1.60 (m, 5H), 1.47 (s, 9H), 1.45 - 1.20 (m, 4H), 1.20 (s, 9H), 1.03 (m, 3H)

[0109] Example 5: Preparation of N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-(4,4-dimethylcyclohexyl)isobutyramide hydrochloride

Chemical formula

[0110] The title compound was obtained through the following steps A, B, C, and D. Step A: Preparation of methyl (2S,4S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-4-(N-(4,4-dimethylcyclohexyl)isobutyramide)pyrrolidine-2-carboxylate

Chemical formula

[0111] (3S,4R)-1-(tert-Butyl)-4-(4-chlorophenyl)pyrrolidine-3-carboxylic acid (0.73 g, 2.58 mmol) obtained in Production Example 5 and methyl (2S,4S)-4-(N-(4,4-dimethylcyclohexyl)isobutylamide)pyrrolidine-2-carboxylate hydrochloride (0.93 g, 2.58 mmol) obtained in Production Example 4 were used, and the title compound (0.49 g, 32%) was obtained in the same manner as in Step A of Example 1. MS [M+H] = 588.5 (M+1) 1 H NMR (400 MHz, CD3OD) δ 7.30 (m, 4H), 4.42 (t, 1H), 4.01 (t, 1H), 3.69 (s, 3H), 3.65 - 3.50 (m, 3H), 3.34 - 3.20 (m, 2H), 3.13 - 3.04 (m, 2H), 2.87 (m, 1H), 2.79 (m, 1H), 2.67 (m, 1H), 2.15 (m, 1H), 1.69 (m, 1H), 1.56 (m, 1H), 1.50 - 1.26 (m, 7H), 1.16 (s, 9H), 1.00 (m, 6H), 0.93 (m, 6H)

[0112] Step B: Preparation of (2S,4S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-4-(N-(4,4-dimethylcyclohexyl)isobutylamide)pyrrolidine-2-carboxylic acid

Chemical formula

[0113] Methyl (2S,4S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-4-(N-(4,4-dimethylcyclohexyl)isobutylamido)pyrrolidine-2-carboxylate (0.49 g, 0.83 mmol) obtained in the above Step A was dissolved in methanol (2.8 mL), cooled to 0 °C, and 6M aqueous sodium hydroxide solution (0.7 mL, 4.2 mmol) was added. The mixture was stirred at room temperature for 16 hours and then concentrated under reduced pressure. Water was added thereto, the pH was adjusted to 4, and the mixture was concentrated under reduced pressure. The mixture was dissolved in dichloromethane, and the solid was removed by filtration. The filtrate was concentrated under reduced pressure to obtain the title compound (0.47 g, 98%). MS [M+H] = 574.4 (M+1) 1 H NMR (400 MHz, CD3OD) δ 7.40 (m, 4H), 4.41 (m, 1H), 4.13 - 3.65 (m, 5H), 3.60 - 3.35 (m, 3H), 2.96 (m, 1H), 2.82 - 2.69 (m, 2H), 2.18 (m, 1H), 1.73 - 1.55 (m, 2H), 1.45 (s, 9H), 1.42 - 1.20 (m, 7H), 1.00 (m, 6H), 0.94 (m, 6H)

[0114] Step C: Preparation of N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-(4,4-dimethylcyclohexyl)isobutylamide

Chemical formula

[0115] (2S,4S)-1-((3S,4R)-1-(tert-Butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-4-(N-(4,4-dimethylcyclohexyl)isobutylamido)pyrrolidine-2-carboxylic acid (0.47 g, 0.82 mmol) obtained in the above step B and morpholine (0.071 mL, 0.82 mmol) were used to obtain the title compound (0.41 g, 78%) in the same manner as in step A of Example 1. MS [M+H] = 643.4 (M+1) 1 H NMR (400 MHz, CD3OD) δ 7.34 (m, 2H), 7.30 (m, 2H), 4.77 (m, 1H), 4.12 (m, 1H), 3.80 - 3.30 (m, 15H), 3.05 (m, 1H), 2.77 (m, 1H), 2.64 (m, 1H), 2.09 (m, 1H), 1.80 - 1.58 (m, 2H), 1.50 - 1.24 (m, 7H), 1.14 (s, 9H), 0.99 (m, 6H), 0.95 (m, 6H)

[0116] Step D: Preparation of N-((3S,5S)-1-((3S,4R)-1-(tert-Butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-(4,4-dimethylcyclohexyl)isobutylamide hydrochloride

Chemical formula

[0117] N-((3S,5S)-1-((3S,4R)-1-(tert-Butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-(4,4-dimethylcyclohexyl)isobutylamide (0.42 g, 0.65 mmol) obtained in the above step C was used to obtain the title compound (0.37 g, 83%) in the same manner as in step B of Example 1. MS [M+H] = 643.4 (M+1) 1 1H NMR (400 MHz, CD3OD) δ 7.44 (m, 4H), 4.81 (m, 1H), 4.22 (m, 1H), 3.90 (m, 1H), 3.80 - 3.40 (m, 14H), 2.99 (m, 1H), 2.82 (m, 1H), 2.68 (m, 1H), 2.16 (m, 1H), 1.80 - 1.60 (m, 2H), 1.40 - 1.20 (m, 7H), 1.47 (s, 9H), 1.05 (m, 6H), 0.96 (m, 6H)

[0118] Example 6: Preparation of N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-(4,4-dimethylcyclohexyl)pivalamide hydrochloride

Chemical Structure

[0119] The title compound was obtained through the following steps A, B, C, and D. Step A: Preparation of methyl (2S,4S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-4-(N-(4,4-dimethylcyclohexyl)pivalamide)pyrrolidine-2-carboxylate

Chemical Structure

[0120] Using (3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carboxylic acid (0.60 g, 2.13 mmol) obtained in Production Example 5 and methyl (2S,4S)-4-(N-(4,4-dimethylcyclohexyl)pivalamide)pyrrolidine-2-carboxylate hydrochloride (0.80 g, 2.13 mmol) obtained in Production Example 3, the title compound (0.63 g, 49%) was obtained in the same manner as in Step A of Example 1. MS [M+H] = 602.5 (M+1) 1 1H NMR (400 MHz, CD3OD) δ 7.34 (m, 4H), 4.47 (t, 1H), 3.98 (m, 1H), 3.80 (m, 1H), 3.71 (s, 3H), 3.60 (m, 2H), 3.48 - 3.34 (m, 3H), 3.17 (m, 1H), 3.10 (m, 1H), 2.63 (m, 1H), 2.16 (m, 1H), 1.73 (m, 1H), 1.58 (m, 1H), 1.52 - 1.38 (m, 3H), 1.36 - 1.20 (m, 4H), 1.23 (s, 9H), 1.17 (s, 9H), 0.97 (s, 3H), 0.94 (s, 3H)

[0121] Process B: Preparation of (2S,4S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-4-(N-(4,4-dimethylcyclohexyl)pivalamide)pyrrolidine-2-carboxylic acid

Chemical formula

[0122] Using methyl (2S,4S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-4-(N-(4,4-dimethylcyclohexyl)pivalamide)pyrrolidine-2-carboxylate (0.63 g, 1.05 mmol) obtained in Process A, the title compound (0.51 g, 83%) was obtained in the same manner as in Process B of Example 5. MS [M+H] = 588.5 (M+1) 11H NMR (400 MHz, CD3OD) δ 7.42 (m, 4H), 4.41 (m, 1H), 4.06 (m, 1H), 3.90 (m, 2H), 3.80 - 3.60 (m, 3H), 3.47 (m, 2H), 2.94 (m, 1H), 2.72 (m, 1H), 2.18 (m, 1H), 1.71 (m, 1H), 1.61 (m, 1H), 1.50 - 1.20 (m, 7H), 1.47 (s, 9H), 1.19 (s, 9H), 0.98 (s, 3H), 0.94 (s, 3H)

[0123] Process C: Preparation of N - ((3S,5S) - 1 - ((3S,4R) - 1 - (tert - butyl) - 4 - (4 - chlorophenyl)pyrrolidine - 3 - carbonyl) - 5 - (morpholine - 4 - carbonyl)pyrrolidin - 3 - yl) - N - (4,4 - dimethylcyclohexyl)pivalamide

Chemical Structure

[0124] (2S,4S) - 1 - ((3S,4R) - 1 - (tert - butyl) - 4 - (4 - chlorophenyl)pyrrolidine - 3 - carbonyl) - 4 - (N - (4,4 - dimethylcyclohexyl)pivalamide)pyrrolidine - 2 - carboxylic acid (0.51 g, 0.87 mmol) obtained in Process B and morpholine (0.076 mL, 0.87 mmol) were used, and the title compound (0.44 g, 77%) was obtained in the same manner as in Process A of Example 1. MS [M + H] = 657.6 (M + 1) 1 1H NMR (400 MHz, CD3OD) δ 7.36 (m, 2H), 7.30 (m, 2H), 4.77 (m, 1H), 4.14 (m, 1H), 3.80 - 3.20 (m, 15H), 3.05 (m, 1H), 2.68 (m, 1H), 2.09 (m, 1H), 1.75 (m, 1H), 1.61 (m, 1H), 1.51 - 1.24 (m, 7H), 1.15 (m, 18H), 0.97 (m, 6H)

[0125] Process D: Preparation of N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-(4,4-dimethylcyclohexyl)pivalamide hydrochloride

Chem.

[0126] Using N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-(4,4-dimethylcyclohexyl)pivalamide (0.44 g, 0.67 mmol) obtained in the above Process C, the title compound (0.39 g, 84%) was obtained in the same manner as in Process B of Example 1. MS [M+H] = 657.6 (M+1) 1 H NMR (400 MHz, CD3OD) δ 7.45 (m, 4H), 4.81 (m, 1H), 4.16 (m, 1H), 4.06 - 3.85 (m, 2H), 3.84 - 3.40 (m, 13H), 2.98 (m, 1H), 2.67 (m, 1H), 2.13 (m, 1H), 1.75 (m, 1H), 1.63 (m, 1H), 1.47 (s, 9H), 1.50 - 1.20 (m, 7H), 1.18 (s, 9H), 0.96 (m, 6H)

[0127] Comparative Example 1: Preparation of N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(2,4-difluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-(4,4-dimethylcyclohexyl)acetamide hydrochloride (A95)

Chem.

[0128] The A95 compound of International Publication No. WO 2008 / 007930 was obtained in the same manner as disclosed therein.

[0129] Comparative Example 2: Production of N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidin-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-(4,4-dimethylcyclohexyl)acetamide hydrochloride (A96) [Chemical formula]

[0130] The A96 compound of International Publication No. WO 2008 / 007930 was obtained in the same manner as disclosed therein.

[0131] Experimental Example 1: Luciferase assay To measure the agonistic activity against MC4R (melanocortin-4 receptor), a cell line that constitutively expresses the luciferase gene (CRE-LUC) under the control of MC4R and CRE (cAMP response element) was established. After producing a mammalian cell expression vector (pCDNA3(Neo)) (manufactured by Invitrogen) containing the MC4R gene, human embryonic kidney (HEK) cell line was transformed using Lipofectamine 2000 (manufactured by Invitrogen) together with a vector (pCRE-Luc) (manufactured by Stratagen) that expresses the luciferase gene (CRE-LUC) under the control of CRE (cAMP response element). The transformed cell line (HEK MC4R-Luc) was cultured for 24 hours in Dulbecco's modified Eagle's medium (DMEM) containing 10% heat-inactivated fetal bovine serum (manufactured by GIBCO / BRL) in a constant temperature incubator at 37°C with 5% CO2. The above cell line was cultured for 4 days in the presence of 10 mL of selection medium (Dulbecco's modified Eagle's medium (DMEM) containing 10% heat-inactivated fetal bovine serum (manufactured by GIBCO / BRL), 100 units / mL of penicillin (manufactured by GIBCO / BRL), 100 units / mL of streptomycin (manufactured by GIBCO / BRL), 800 μg / mL of geneticin (G418) (manufactured by GIBCO / BRL)). The process of removing cells killed by the selection medium by replacing the medium with 10 mL of fresh selection medium was repeated 3 times, once every 4 days. Individual colonies formed by the finally selected and proliferated clones were transferred under a microscope to a 24-well cell culture plate containing 1 mL of selection medium per well and cultured for 4 days. Forskolin (manufactured by SIGMA) was treated to a final concentration of 10 μM and cultured for 5 hours in a constant temperature incubator at 37°C with 5% CO2. 50 μL of Bright-Glo luciferase reagent (manufactured by Promega) was treated to each well, left at room temperature for 15 minutes, and then the luminescence of each well was measured using a luminometer (manufactured by Victor). Clones showing luminescence more than 100 times the basal value by the treatment of forskolin were selected and used to measure the MC4R agonistic activity of each compound.

[0132] HEK MC4R-Luc cells were added to each well of a 96-well luminometer cell culture plate (Costar) at a density of 2.5×10 4 cells in 100 μL of medium and cultured in a 37°C incubator with 6% CO2 for 18 hours. MCR agonists diluted to each concentration step using the above medium were treated so that the final DMSO concentration did not exceed 1%, and cultured in a 37°C incubator with 6% CO2 for 5 hours. Each well was treated with 50 μL of Bright-Glo luciferase reagent (Promega), left at room temperature for 5 minutes, and then the luminescence of each well was measured using a luminometer (Victor). The luminescence induced by the agonist diluted at each concentration step was converted to a relative % value relative to the amount shown by the 0 μM NDP-α-MSH treatment. EC 0.5 MSH is the concentration that induces 50% of the maximum luminescence that can be induced by NDP-α-MSH, and EC 50 is expressed as the concentration that induces 50% of the maximum luminescence that can be induced by each agonist. The above measurements were made using statistical software (Prizm).

[0133] The MC4R agonistic ability of each compound obtained from the above experiment was measured and the results are shown in Table 1 in units of EC 50 (nM).

Table 1

[0134] As shown in Table 1 above, among the melanocortin receptors well-known in vivo, with respect to the melanocortin-4 receptor (MC4R) involved in energy metabolism and body weight control in vivo, it was confirmed that the compounds of the examples had superior MC4R agonistic ability compared to the compounds of the comparative examples (A95 and A96).

[0135] Experimental Example 2: cAMP assay The melanocortin receptor is a type of G protein-coupled receptor (GPCR). The main role of G protein is to activate secondary messengers that control cellular responses to many physiological stimuli through signal transduction. MC4R is a Gs-coupled receptor. When MC4R interacts with an agonist, adenylate cyclase (AC) is activated, and it is known that the concentration of cyclic AMP (cAMP), one of the intracellular secondary messengers, increases. Therefore, the activity of the melanocortin receptor can be evaluated by measuring the generation of cAMP signals.

[0136] After establishing cAMP hunter Gs-coupled receptor cell lines (CHO-K1 cell lines) overexpressing each of MC1R, MC3R, MC4R, and MC5R so that the increase in intracellular cAMP levels due to agonist response can be measured, the cells were inoculated into each well of a white cell culture plate and cultured in a 37 °C constant temperature incubator with 5% CO2 for 24 hours. After culturing, the medium was removed, and 15 μL of 2:1 HBBS / 10 mM HEPES:cAMP XS+Ab reagent was added. After adding 5 μL of the sample diluted 4-fold with the buffer, the vehicle concentration was set to 1%, and MC4R agonist compounds diluted at each step concentration were added and reacted at 37 °C for 30 minutes. The activity (%) of each agonist compound was expressed as 100%×(average RLU value of the sample - average RLU value of the vehicle control) / (average RLU value of the maximum control - average RLU value of the vehicle control), and the said value was analyzed with the CBIS data analysis suite (manufactured by ChemInnovation, CA).

[0137] The agonist ability of the melanocortin receptor of each compound obtained from the said experiment was measured in EC 50 (nM) units, and the results are shown in Table 2 below.

Table 2

[0138] As shown in Table 2 above, among the melanocortin receptors well-known in vivo, with respect to the melanocortin-4 receptor (MC4R) involved in energy metabolism and body weight control in vivo, it was confirmed that the compounds of the examples have a better receptor agonist ability than the compounds of Comparative Examples (A95 and A96).

[0139] Experimental Example 3: β-Arrestin Assay Melanocortin receptors are a type of G protein-coupled receptor (GPCR) and control various physiological reactions by transmitting signals from many neurotransmitters. When the GPCR is phosphorylated, β-arrestin binds to the phosphorylated part of the receptor and plays an important role in activating various intracellular signal transduction pathways through interactions with other proteins. When the melanocortin receptor interacts with an agonist, β-arrestin is mobilized and is known to be involved in the β-arrestin-mediated signal transduction pathway. Therefore, the activity of the melanocortin receptor can be evaluated by measuring β-arrestin.

[0140] A Pathhunter eXpress β-arrestin cell line (U2OS cell line) co-expressing Prolink (PK)-tagged MC1R, MC3R, MC4R, MC5R and enzyme acceptor (EA)-tagged β-arrestin was established. When the MCR-PK portion of this cell line is activated, β-arrestin-EA is mobilized, and the oxygen acceptor (EA), which is a β-galactosidase enzyme fragment, interacts with Prolink (PK). The activated enzyme hydrolyzes the substrate by β-galactosidase activity to generate a chemiluminescence signal, and its activity can be measured. After culturing the Pathhunter eXpress β-arrestin cell line (U2OS cell line), the cells were inoculated into each well of a cell culture plate and cultured in a constant temperature incubator at 37 °C in the presence of 5% CO2 for 48 hours. After culturing, 5 μL of a sample diluted 5-fold with buffer was added, the vehicle concentration was set to 1%, and an MC4R agonist compound diluted at each step concentration was added, followed by reacting at 37 °C for 90 minutes. The activity (%) of each agonist compound is represented by 100% × (average RLU value of the sample - average RLU value of the vehicle control) / (average maximum value of the control ligand - average RLU value of the vehicle control), and the said value was analyzed by the CBIS data analysis suite (manufactured by ChemInnovation, CA).

[0141] The agonist ability of the melanocortin receptor of each compound obtained from the said experiment was measured in EC 50 (nM) units, and the results are shown in Table 3 below.

Table 3

[0142] As shown in Table 3 above, among the melanocortin receptors well-known in vivo, with respect to the melanocortin-4 receptor (MC4R) involved in energy metabolism and body weight control in vivo, it was confirmed that the compounds of the examples had superior receptor activation ability compared to the compounds of the comparative examples (A95 and A96).

[0143] Experimental Example 4: Binding Affinity There are five subtypes of melanocortin receptors (MCR) in the body, and MC4R, which is subtype 4, is known to be involved in energy metabolism and body weight control. In the case of other MCR subtypes, since they are involved in the regulation of various functions in the body such as skin pigmentation, energy homeostasis, and exocrine function, ensuring the selectivity of MC4R agonist compounds for MC4R is very important for preventing possible side effects in the future. Therefore, the receptor binding ability of MC4R agonists to each MCR subtype was measured.

[0144] After establishing CHO-K1 cell lines expressing human recombinant MC1R, HEK-293 cell lines expressing MC3R, MC4R, and MC5R, membranes were recovered from each cell line. In a 96-well cell culture plate, 3 μg of MC1 membrane per well and 0.04 nM 125 I-NDP-α-MSH were added and reacted at 37 °C for 2 hours. 3 μg of MC3R, MC5 membranes and 0.035 nM 125 I-NDP-α-MSH were reacted at 37 °C for 1 hour, and 3.12 μg of MC4R membrane and 0.02 nM 125 I-NDP-α-MSH were reacted at 37 °C for 2 hours. At this time, 25 mM HEPES-KOH adsorption buffer (pH 7.0) containing MCR agonists diluted at each concentration step was added to each well and reacted. The reacted solution was transferred to a filter, washed with adsorption buffer, and then radioactivity was measured. The value obtained by subtracting the non-specific binding amount in the presence of 1 μM (MC1R), 3 μM (MC3R, MC4R, MC5R) NDP-α-MSH from each total binding amount was 125 regarded as the specific binding amount of 125 I-NDP-α-MSH. The degree to which the specific binding of 50 I-NDP-α-MSH was inhibited by agonists diluted at each concentration step was measured. IC 125 was expressed as the concentration of each agonist that inhibits 50% of the specific binding of

[0145] The results of measuring the binding of each compound obtained from the above experiment to melanocortin receptors in units of Ki (nM) are shown in Tables 4 and 5.

Table 4

Table 5

[0146] As shown in Tables 4 and 5 above, among the melanocortin receptors well known in vivo, with respect to the melanocortin-4 receptor (MC4R) involved in energy metabolism and body weight control in vivo, it was confirmed that the compounds of the examples have a receptor binding ability superior to that of the compounds of the comparative examples (A95 and A96).

Claims

1. The following formula (1): 【Chemical 1】 (In the formula, R1 is C 2 -C 5 alkyl, R2 is halo, R3 is hydrogen or halo, R4 is C 1 -C 3 alkyl, and n is an integer of 1 or 2, provided that when R2 is chlorine and R3 is hydrogen, n is 2.), or a pharmaceutically acceptable salt or isomer thereof, wherein the isomer includes cis or trans isomers, R or S isomers, racemates, or diastereoisomers, a compound, or a pharmaceutically acceptable salt or isomer thereof.

2. R1 is C 2 -C 4 The compound according to claim 1, wherein R1 is alkyl, or a pharmaceutically acceptable salt or isomer thereof.

3. The compound of formula (1) according to claim 2, or a pharmaceutically acceptable salt or isomer thereof, selected from the following group: N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(2,4-difluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)pivalamide; N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(2,4-difluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)isobutyramide; N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chloro-2-fluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)isobutyramide; N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chloro-2-fluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)pivalamide; N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-(4,4-dimethylcyclohexyl)isobutyramide; and N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-(4,4-dimethylcyclohexyl)pivalamide.

4. The compound according to claim 1, or a pharmaceutically acceptable salt or isomer thereof, wherein the pharmaceutically acceptable salt is selected from the group consisting of hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrobromic acid, and hydroiodic acid.

5. The compound according to claim 4, or a pharmaceutically acceptable salt or isomer thereof, wherein the pharmaceutically acceptable salt is a hydrochloride.

6. A melanocortin-4 receptor agonist pharmaceutical composition comprising the compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt or isomer thereof, and a pharmaceutically acceptable carrier.

7. The pharmaceutical composition according to claim 6, which is for the prevention or treatment of obesity.

8. The pharmaceutical composition according to claim 6, which is for the prevention or treatment of diabetes.

9. The pharmaceutical composition according to claim 6, which is for the prevention or treatment of inflammation.

10. The pharmaceutical composition according to claim 6, which is for the prevention or treatment of erectile dysfunction.

11. The compound of formula (1) according to claim 3, or a pharmaceutically acceptable salt or isomer thereof, wherein the compound is N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(2,4-difluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)pivalamide.

12. The compound of formula (1) according to claim 3, or a pharmaceutically acceptable salt or isomer thereof, wherein the compound is N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(2,4-difluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)isobutyramide.

13. The compound of formula (1) is N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chloro-2-fluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)isobutyramide, the compound according to claim 3, or a pharmaceutically acceptable salt or isomer thereof.

14. The compound of formula (1) is N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chloro-2-fluorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)pivalamide, the compound according to claim 3, or a pharmaceutically acceptable salt or isomer thereof.

15. The compound of formula (1) is N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-(4,4-dimethylcyclohexyl)isobutyramide, the compound according to claim 3, or a pharmaceutically acceptable salt or isomer thereof.

16. The compound of formula (1) is N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-(4,4-dimethylcyclohexyl)pivalamide, the compound according to claim 3, or a pharmaceutically acceptable salt or isomer thereof.

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