Use of melanocortin-4 receptor agonists in the prevention or treatment of rare genetic obesity

Abstract

The present invention relates to the use of a compound of formula 1 or a pharma- ceutically acceptable salt thereof for preventing or treating a rare genetic obesity disease associated with the melanocortin-4 receptor (MC4R) pathway, in particular a rare genetic obesity disease associated with proopiomelanocortin (POMC) deficiency.

Description

[Technical field]

[0001] The present invention relates to the use of a compound of formula 1 or a pharma- ceutically acceptable salt thereof for preventing or treating a rare genetic obesity disorder associated with the melanocortin-4 receptor (MC4R) pathway, in particular a rare genetic obesity disorder associated with proopiomelanocortin (POMC) deficiency: [ka] (In formula 1, R1 is a C2 to C5 alkyl). [Background technology]

[0002] Melanocortin receptors (MCRs) are a type of G protein-coupled receptor (GPCR), and the main role of G proteins is to activate second messengers and regulate cellular responses to many physiological stimuli via signal transduction. Five melanocortin receptors have been identified to date. MC1R is expressed in melanocytes and macrophages, where it determines skin and hair color by regulating melanin pigment. MC2R is expressed in the adrenal gland and adipose tissue, where its mediating function in regulating corticotropin-mediated secretion of adrenal corticosteroids in the adrenal gland is well known. MC3R, MC4R, and MC5R are expressed in the brain and nerve terminals and are thought to mediate the central nervous actions of melanocortin peptides, which are manifested in effects on behavior, learning, memory, appetite, and neuronal development and regeneration. To date, MC3R is known to be involved in erectile dysfunction and inflammatory responses, and MC4R is known to be involved in obesity and diabetes, and research into the specificity of each receptor is being actively conducted (MacNeil DJ et al., Eur J Pharmacol. 2002, 450, 93). As a result, genetic studies in humans with active obesity have shown that MC4R is deeply involved, and genetically modified mice (knockout mice) in which MC4R has been deleted have been shown to develop obesity through overeating, indicating that this receptor plays an important role in appetite regulation ((Lu D, Willard D et al., Nature 1994, 371 (6500), 799; Huszar D et al., Cell 1997, 88 (1), 131; Hinney A et al., J Clin Endocrinol Metab 1990, 84 (4), 1483).

[0003] In contrast, existing obesity treatment drugs are mainly appetite suppressants that act on the central nervous system, and most of them are drugs that regulate the action of neurotransmitters (e.g., phentermine, mazindol, lorcaserin, fluoxetine, and sibutramine). However, these neurotransmitter modulators have a wide range of effects on various physiological functions other than appetite suppression through many subtype receptors. Therefore, the above drugs have the disadvantage of lacking selectivity for each subtype, and therefore cause various side effects when administered for a long period of time. In contrast, melanocortin antagonists, which are neuropeptides rather than neurotransmitters, have the advantage of being able to induce weight loss by appetite suppression without affecting other physiological functions in MC4R gene knockout (KO) mice, in which all other functions other than energy metabolism are normal. Summary of the Invention

[0004] Problem to be solved by the invention The present invention provides a compound of formula 1 or a pharma- ceutically acceptable salt thereof for use in the prophylaxis or treatment of a rare genetic obesity disorder associated with the melanocortin-4 receptor (MC4R) pathway, particularly a rare genetic obesity disorder associated with proopiomelanocortin (POMC) deficiency: [ka] (In formula 1, R1 is a C2 to C5 alkyl).

[0005] Means for solving the problem The present invention provides a medicament for preventing or treating a rare genetic obesity disease associated with proopiomelanocortin (POMC) deficiency, comprising a therapeutically effective amount of a compound of formula 1 or a pharma- ceutical acceptable salt thereof: [ka] (In formula 1, R1 is a C2 to C5 alkyl).

[0006] The present invention also provides a pharmaceutical composition for preventing or treating a rare genetic obesity disease associated with proopiomelanocortin (POMC) deficiency, comprising a therapeutically effective amount of a compound of formula 1 or a pharma- ceutical acceptable salt thereof, together with a pharma- ceutical acceptable carrier.

[0007] The present invention further provides a method for preventing or treating a rare genetic obesity disease associated with proopiomelanocortin (POMC) deficiency, comprising administering to a subject in need of preventing or treating a rare genetic obesity disease a therapeutically effective amount of a compound of formula 1 or a pharma- ceutically acceptable salt thereof.

[0008] The present invention also provides the use of a compound of formula 1, or a pharma- ceutically acceptable salt thereof, for preventing or treating a rare genetic obesity disorder associated with proopiomelanocortin (POMC) deficiency.

[0009] The present invention will now be described in more detail.

[0010] According to one aspect of the present invention, there is provided a medicament for preventing or treating rare genetic obesity associated with proopiomelanocortin (POMC) deficiency, comprising a therapeutically effective amount of a compound of formula 1 or a pharma- ceutical acceptable salt thereof.

[0011] According to another aspect of the present invention, there is also provided a pharmaceutical composition for preventing or treating rare genetic obesity associated with proopiomelanocortin (POMC) deficiency, comprising a therapeutically effective amount of a compound of formula 1 or a pharma- ceutical acceptable salt thereof, together with a pharma- ceutical acceptable carrier.

[0012] In one embodiment according to the invention, 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-((1s,4R)-4-methylcyclohexyl)isobutyramide of formula 2 below: [ka]

[0013] In another embodiment according to the invention, the pharma-ceutically acceptable salt includes, but is not limited to, an acid addition salt formed with an inorganic acid such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrobromic acid, hydroiodic acid, an organic carboxylic acid 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 a sulfonic acid such as methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, or naphthalenesulfonic acid. In another embodiment according to the invention, the pharma-ceutically acceptable salt can be selected from the group consisting of hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrobromic acid, and hydroiodic acid. In some embodiments, the pharma-ceutically acceptable salt is a hydrochloride salt.

[0014] In another embodiment according to the present invention, the hydrochloride salt of the compound of formula 1 can be prepared according to the following scheme 1. However, a person skilled in the art to which the present invention pertains can prepare the compound of formula 1 by various methods based on the structure of formula 1. [Scheme 1] [ka]

[0015] In the above Scheme 1, R2 is a C1-C5 alkyl group; R3 is a C3-C8 cycloalkyl unsubstituted or substituted with 1 or 2 C1-C5 alkyl groups; R4 and R5 are each independently hydrogen or halogen.

[0016] According to the present invention, the compound of formula 1 can effectively prevent or treat rare genetic obesity disease associated with proopiomelanocortin (POMC) deficiency. Proopiomelanocortin (POMC) is a precursor polypeptide with 241 amino acid residues, which is synthesized in the anterior and intermediate pituitary glands and acts as a precursor of several peptide hormones such as melanocyte-stimulating hormone (MSH), endorphins, and corticotrophins. Alpha-MSH acts upstream of melanocortin-4 receptor (MC4R) as a ligand of MC4R, and when the Pomc gene encoding it is defective and not normally expressed, it can cause severe early-onset obesity.

[0017] In another embodiment according to the present invention, a "therapeutically effective amount" in an individual subject means an amount sufficient to obtain the above-mentioned pharmacological effect, i.e., a therapeutic effect, and the amount of the compound depends on the condition of the subject and its severity, the method of administration, and the age of the subject to be treated, but can be determined by a person skilled in the relevant technical field based on his or her knowledge.

[0018] In another embodiment according to the invention, a therapeutically effective dosage of a compound of formula 1, e.g., the dosage required to treat an adult, generally ranges from about 0.1 to 500 mg / day, depending on the frequency and intensity of administration. For intramuscular or intravenous administration to adults, a total dosage of about 0.1 to 300 mg / day divided into single doses is usually sufficient, although higher daily dosages may be desirable in some patients.

[0019] The "pharmaceutical composition" of the present invention may contain other chemical components such as carriers, diluents, excipients, etc., in addition to the active compound according to the present invention. Thus, the pharmaceutical composition may contain a pharma- ceutically acceptable carrier, diluent, excipient, or a combination thereof, as necessary. The pharmaceutical composition facilitates administration of the compound to a living body. There are various methods for administering the compound, including, but not limited to, oral, injection, aerosol, parenteral, and topical administration.

[0020] As used herein, "carrier" refers to a compound that facilitates the introduction of a compound into cells or tissues.For example, dimethylsulfoxide (DMSO) is a common carrier that facilitates the introduction of many organic compounds into cells or tissues of living organisms.

[0021] As used herein, "diluent" is defined as a compound that is diluted in water to stabilize the biologically active form of the compound of interest and dissolve the compound. Salts dissolved in buffered solutions are used as diluents in the art. A commonly used buffer is phosphate buffered saline, which mimics the salt form of human body fluids. Buffered diluents rarely alter the biological activity of a compound, since buffer salts can adjust the pH of the solution at low concentrations.

[0022] As used herein, "pharmacologically acceptable" means a property that does not abrogate the biological activity and physical properties of a compound.

[0023] In the present invention, compound can be formulated into various pharmaceutical dosage forms according to purpose.When preparing pharmaceutical composition according to the present invention, active ingredient, specifically, the compound of formula 1 or its pharmaceutically acceptable salt, is mixed with various pharmaceutically acceptable carriers that can be selected according to the preparation to be prepared.For example, pharmaceutical composition according to the present invention can be formulated as injection preparation, oral preparation, etc. according to purpose.

[0024] The compounds of the present invention can be formulated by known methods using known pharmaceutical carriers and excipients, and can be introduced in unit dosage form or multi-dose containers. The formulations can be in the form of solutions, suspensions, or emulsions in oily or aqueous media, and can contain conventional dispersing, suspending, or stabilizing agents. The formulations can be in the form of dry powders, for example, dissolved in sterilized pyrogen-free water prior to use. The compounds of the present invention can also be formulated into suppository form using conventional suppository bases, such as cocoa butter or other glycerides. Solid dosage forms for oral administration can be capsules, tablets, pills, granules, and tablets, with capsules and tablets being particularly useful. Tablets and pills are preferably prepared with enteric coatings. Solid dosage forms can be prepared by mixing the compounds of the present invention with one or more inert diluents, such as sucrose, lactose, starch, and carriers, such as lubricants, disintegrants, binders, and the like, for example magnesium stearate. The composition may also be formulated as a transdermal dosage form, such as, for example, a lotion, ointment, gel, cream, patch, or spray. In another embodiment according to the invention, the drug or pharmaceutical composition may be in an oral formulation.

[0025] As used herein, "prevention" means reducing or eliminating the chance of contracting a disease.

[0026] As used herein, "treatment" means to halt, slow or alleviate the progression of the disease when used on a subject who exhibits symptoms of the disease.

[0027] Effect of the Invention The drug or pharmaceutical composition according to the present invention can effectively prevent or treat rare genetic obesity diseases associated with the melanocortin-4 receptor (MC4R) pathway, specifically, rare genetic obesity diseases associated with proopiomelanocortin (POMC) deficiency. [Brief description of the drawings]

[0028] [Figure 1]1 is a graph showing the results of the anti-obesity effect in a POMC heterozygous deficient mouse model on a high-fat diet (HFD). [Diagram 2] 1 is a graph showing the results of the anti-obesity effect in a POMC homozygous deficient mouse model on a high-fat diet (HFD). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0029] The present invention will be described in more detail below with reference to examples. However, these examples are merely intended to illustrate one or more embodiments, and the scope of the present invention is not limited thereby.

[0030] Preparation Example: Synthesis of N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)isobutyramide hydrochloride [ka]

[0031] The title compound was obtained by steps A, B, C, and D below.

[0032] Step A: Preparation of methyl (2S,4S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-4-(N-((1s,4R)-4-methylcyclohexyl)isobutyramido)pyrrolidine-2-carboxylate

[0033] Methyl (2S,4S)-4-(N-((1s,4R)-4-methylcyclohexyl)isobutylamido)pyrrolidine-2-carboxylate hydrochloride (28.7 g, 82.73 mmol) obtained from Preparation Example 1, (3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carboxylic acid (24.5 g, 86.87 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (22.2 g, 115.83 mmol), and 1-hydroxybenzotriazole hydrate (15.7 g, 115.83 mmol) obtained from Preparation Example 2 were dissolved in N,N'-dimethylformamide (400 ml), and N,N'-diisopropylethylamine (72.0 ml, 413.66 mmol) was slowly added. After stirring at room temperature for 16 hours, the reaction solvent was concentrated under reduced pressure, 0.5N aqueous sodium hydroxide solution was added, and the mixture was extracted twice with ethyl acetate. The organic layer was washed twice with aqueous sodium chloride solution and water, and then dry filtered over anhydrous magnesium sulfate. The filtrate was concentrated under reduced pressure and purified by column chromatography to obtain the title compound (41.19 g, 87%). MS [M+H] = 575 (M+1)

[0034] Step B: Preparation of (2S,4S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-4-(N-((1s,4R)-4-methylcyclohexyl)isobutyramido)pyrrolidine-2-carboxylic acid

[0035] Methyl (2S,4S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-4-(N-((1s,4R)-4-methylcyclohexyl)isobutyramido)pyrrolidine-2-carboxylate (39.4 g, 68.62 mmol) obtained from step A above was dissolved in methanol (450 ml), and then 6N sodium hydroxide (57.2 ml, 343.09 mmol) was added. After stirring at room temperature for 16 hours and adjusting the pH to about 5 with 6N hydrochloric acid, the reaction solution was concentrated under reduced pressure. The concentrate was dissolved in dichloromethane, and the insoluble solid was filtered through filter paper. The filtrate was concentrated under reduced pressure to give the crude product (38.4 g, 99%), which was used in the next step without purification. MS [M+H] = 561 (M+1)

[0036] 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-((1s,4R)-4-methylcyclohexyl)isobutyramide

[0037] (2S,4S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-4-(N-((1s,4R)-4-methylcyclohexyl)isobutyramide)pyrrolidine-2-carboxylic acid (38.4 g, 68.60 mmol) obtained from step B above, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (18.4 g, 96.04 mmol), and 1-hydroxybenzotriazole hydrate (13.0 g, 96.04 mmol) were dissolved in N,N'-dimethylformamide (200 ml), and then morpholine (5.9 ml, 68.80 mmol) and N,N'-diisopropylethylamine (59.7 ml, 343.02 mmol) were added slowly in sequence. After stirring at room temperature for 16 hours, the reaction solution was concentrated under reduced pressure, then 0.5N aqueous sodium hydroxide solution was added, and extracted twice with ethyl acetate. The organic layer was washed twice with aqueous sodium chloride solution and water, and then dried over anhydrous magnesium sulfate. The filtrate was concentrated under reduced pressure and purified by column chromatography to obtain the title compound (37.05 g, 86%). MS [M+H] = 630 (M+1)

[0038] 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-((1s,4R)-4-methylcyclohexyl)isobutyramide hydrochloride

[0039] N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)isobutyramide hydrochloride (5.0 g, 7.95 mmol) obtained from step C above was dissolved in ethyl acetate (50 ml), and then 2N hydrochloric acid-ethyl acetate solution (3.97 ml, 15.89 mmol) was slowly added. After stirring at room temperature for 30 minutes, the reaction solvent was concentrated under reduced pressure. The resulting crude solid was purified by trituration with hexane and diethyl ether to give the title compound (5.23 g, 99%). MS [M+H] = 630 (M+1)

[0040] 1 H NMR (500 MHz, CD3OD) δ 7.49-7.44 (m, 4H), 4.83 (m, 1H), 4.23-4.20 (m, 1H), 3.95-3.91 (m, 2H), 3.79-3.47 (m, 14H), 3.03-3.00 (m, 1H), 2.86-2.82 (m, 1H), 2.73-2.67 (m, 1H), 2.20-2.14 (m, 1H), 1.97 (m, 1H), 1.80-1.62 (m, 5H), 1.50 (s, 9H), 1.44-1.27 (m, 3H), 1.06-1.04 (m, 9H)

[0041] Example: Pomc knockout mouse model experiment

[0042] Proopiomelanocortin (POMC) is a precursor polypeptide of alpha-MSH, a ligand of the melanocortin-4 receptor (MC4R), and acts upstream of the melanocortin-4 receptor. POMC knockout mice, in which the proopiomelanocortin protein is not properly expressed, are a model reflecting POMC-deficient obesity, a genetic obesity disease, and the mice show abnormal metabolic disease-related phenotypes such as excessive appetite and severe obesity.

[0043] Pomc heterozygous knockout mice, which do not normally express the Pomc gene encoding proopiomelanocortin (POMC), were fed a 60 kcal% high-fat diet (diet containing 60 kcal% fat), and N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)isobutyramide hydrochloride (hereinafter referred to as the "test compound") obtained from the preparation example was administered at a dose of 10 mg / kg or 30 mg / kg for about 3 weeks to examine the anti-obesity effect, and the results are shown in Figure 1.

[0044] Pomc homozygous knockout mice, which do not express the Pomc gene encoding proopiomelanocortin (POMC), were fed a 45 kcal% high-fat diet (diet containing 45 kcal% fat) and administered the test compound at a dose of 10 mg / kg or 30 mg / kg for approximately 10 days to examine the anti-obesity effect. The results are shown in Figure 2.

[0045] As can be seen from Figures 1 and 2, it can be confirmed that the test compound of the present invention exhibits excellent anti-obesity effects.

Claims

1. 1. A composition for preventing or treating a rare genetic obesity disorder associated with proopiomelanocortin (POMC) deficiency, comprising a therapeutically effective amount of a compound of formula 1 or a pharmaceutically acceptable salt thereof: 【Chemistry 1】 (wherein R1 is a C2-C5 alkyl).

2. The composition according to claim 1, wherein 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-((1s,4R)-4-methylcyclohexyl)isobutyramide of formula 2 below. 【Chemistry 2】

3. 10. The composition of claim 1, 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.

4. The composition of claim 1 in an oral dosage form.

5. The composition of claim 1, wherein the compound is formulated as a pharmaceutical composition together with a pharmaceutically acceptable carrier.

6. 10. Use of a compound of formula 1 as defined in claim 1 or a pharmaceutically acceptable salt thereof for the preparation of a composition for preventing or treating a rare genetic obesity disorder associated with proopiomelanocortin (POMC) deficiency.

7. The composition of claim 1, wherein R1 is ethyl.

8. The composition of claim 1, wherein R1 is n-propyl.

9. The composition of claim 1, wherein R1 is isopropyl.

10. The composition of claim 1, wherein R1 is n-butyl.

11. The composition of claim 1, wherein R1 is isobutyl.

12. The composition of claim 1, wherein R1 is sec-butyl.

13. The composition of claim 1, wherein R1 is tert-butyl.

14. The compound of claim 1, N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)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-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)propionamide; and N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)pivalamide 10. The composition of claim 1, selected from the group consisting of:

15. The composition of claim 14, wherein the compound is N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)isobutyramide.

16. The composition of claim 14, wherein the compound is N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)propionamide.

17. The composition of claim 14, wherein the compound is N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)pivalamide.

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