Crystalline form iv of melanocortin receptor agonist compound and preparation method therefor

Abstract

To provide a crystalline form IV of a compound represented by a chemical formula 1, a method for preparing the same, and a pharmaceutical composition comprising the same.SOLUTION: A crystalline form IV of a compound represented by a chemical formula 1 of the present invention may be characterized by XRD patterns, DSC profiles, and / or TGA profiles.SELECTED DRAWING: None

Description

Technical Field

[0001] This application claims the benefit of priority based on Korean Patent Application No. 10-2020-0142400, filed on October 29, 2020, and all the contents disclosed in the document of the Korean Patent Application are incorporated herein by reference in their entirety.

[0002] The present invention relates to a crystalline form IV of a novel compound showing excellent enhancing activity against melanocortin receptors, a method for producing the same, and a pharmaceutical composition containing the same.

Background Art

[0003] Leptin protein is a hormone secreted by adipocytes, and its secretion amount increases as the body fat content increases. By regulating the functions of various neuropeptides produced in the hypothalamus, it regulates various biological functions including appetite, body fat content, and energy metabolism (Schwartz, et al., Nature 404, 661-671 (2000)). The signal transduction of appetite and body weight regulation by leptin protein is carried out by the regulation of various factors downstream, and the most typical ones are melanocortin, AgRP (agouti-related peptide), and neuropeptide Y (NPY) hormones.

[0004] As a result of excessive calories in the body, when the concentration of leptin in the blood increases, 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 alpha-MSH (melanocyte stimulating hormone) is produced from POMC neurons, and this hormone ultimately induces a decrease in appetite as an agonist of the melanocortin-4 receptor (MC4R) in secondary neurons. On the other hand, as a result of calorie deficiency, when the concentration of leptin decreases, the expression of AgRP, which is an MC4R antagonist, increases, and the expression of NPY also increases, ultimately promoting appetite. That is, in response to changes in leptin, the alpha-MSH hormone and the AgRP hormone play roles of enhancement and antagonism against MC4R, respectively, and are involved in appetite regulation.

[0005] Alpha-MSH hormone binds to three MCR subtypes in addition to MC4R, inducing various physiological responses. To date, five MCR subtypes have been identified. Among them, MC1R is mainly expressed in skin cells and is involved in skin pigmentation regulation. MC2R is mainly expressed in the adrenal gland and is known to be involved in the production of glucocorticoid hormone. 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 body fat storage efficiency. MC5R, which is expressed in various tissues, is known to regulate exocrine function (Wikberg, et al., Pharm Res 42(5)393-420(2000)). In particular, activation of the MC4R receptor has been proven to be a major target for the development of anti-obesity agents, as it shows the effect of efficiently reducing body weight by inducing a decrease in appetite and an increase in energy metabolism (Review: Wikberg, Eur.J.Pharmacol 375,295-310(1999)); Wikberg, et al., Pharm Res 42(5)393-420(2000); Douglas et al., Eur J Pharm 450,93-109(2002); O’Rahilly et al., Nature Med 10,351-352(2004)).

[0006] In appetite and body weight regulation, the role of MC4R was primarily demonstrated by experiments using animal models with abnormal expression of the agouti protein (agouti mouse). In the case of agouti mice, genetic mutations cause the agouti protein to be expressed at high concentrations in the central nervous system, where it acts as an antagonist of MC4R in the hypothalamus, inducing obesity (Yen, TT et al., FASEB J. 8, 479-488 (1994); Lu D., et al. Nature 371, 799-802 (1994)). Subsequent research findings have observed that in hypothalamic neurons, AgRP (agouti-related peptide), which is similar to the agouti protein, is actually expressed, and these are also known to be involved in appetite regulation as antagonists of MC4R (Shutter, et al., Genes Dev., 11, 593-602 (1997); Ollman, et al. Science 278, 135-138 (1997)).

[0007] When alpha-MSH, an in vivo MC4R agonist, is administered intracerebrally to animals, it shows an effect of reducing appetite. In contrast, when animals are treated with SHU9119 (peptide) or HS014 (peptide), which are MC4R antagonists, a phenomenon of increasing appetite has been observed (Kask et al., Biochem. Biophys. Res. Comm. 245, 90-93 (1998)). Moreover, in animal tests using Melanotan II (MTII, Ac-Nle-c[Asp-His-DPhe-Arg-Trp-Lys]-NH2) and its similar agonist HP228, after intracerebral, intraperitoneal or subcutaneous administration, effects such as appetite suppression, weight loss, and increased energy metabolism have been confirmed (Thiele T.E., et al. Am J Physiol 274(1 Pt 2), R248-54(1998); Lee M.D., et al. FASEB J 12, A552(1998); Murphy B., et al. J Appl Physiol 89, 273-82(2000)). Conversely, administration of the representative SHU9119 to animals shows significant and continuous feed intake and weight gain, providing pharmacological evidence that MCR agonists can be used as anti-obesity agents. The significant appetite-reducing effect that appears during MTII administration does not appear in MC4R KO (knock-out) mice, and this experimental result also proves that the appetite-reducing effect is mainly carried out by the activation of MC4R (Marsh, et al., Nat Genet 21, 119-122(1999)).

[0008] To date, the main types of obesity treatment agents developed are appetite suppressants that act on the central nervous system. Among them, most are drugs that regulate the action of neurotransmitters. Examples include phentermine and mazindol, which are noradrenalin agents, and fluoxetine and sibutramine, which are serotonergic agents. However, in the case of the neurotransmitter regulators, due to numerous subtype receptors, in addition to inhibiting appetite, they also have a wide range of effects on various physiological functions. Therefore, in the case of the regulators, there is a lack of selectivity for each subtype, and when administered for a long time, there are significant drawbacks accompanied by various side effects.

[0009] On the other hand, melanocortin agonists are neuropeptides rather than neurotransmitters. From the point that all other functions except energy metabolism are normal in MC4R gene KO mice, they have the advantage as an action site in that they can induce only weight loss due to appetite inhibition without affecting other physiological functions. In particular, since the receptor belongs to the G-protein coupled receptor (GPCR), which is the most successful category among the new drug action sites developed to date, ensuring selectivity for subtype receptors is relatively easy, which is a significant difference from the existing action sites.

[0010] As an example of utilizing such a melanocortin receptor as an action site, International Publication Nos. WO2008 / 007930 and WO2010 / 056022 disclose compounds as agonists of the melanocortin receptor.

[0011] In addition, the inventors of the present invention have intensively conducted research and invented a novel compound represented by the following Chemical Formula 1, which has excellent selective enhancing activity against melanocortin receptors, particularly melanocortin-4 receptor (MC4R), and a method for producing the same (Korean Application No. 10-2019-0141649 (filed on November 7, 2019)).

[0012]

Chemical Formula

[0013] On the other hand, the crystal structure of a pharmaceutically active ingredient sometimes affects the chemical stability of the drug. Different crystallization conditions and storage conditions may change the crystal structure of the compound and sometimes cause production accompanied by different forms of crystal forms. Generally, amorphous drug products do not have a regular crystal structure and sometimes have other defects such as poor product stability, smaller particle size, difficult filtration, easy aggregation, and poor fluidity. Therefore, it is necessary to improve various physical properties of the product. Thus, it is necessary to study a crystal structure having high purity and good chemical stability for one compound.

Prior Art Documents

Patent Documents

[0014]

Patent Document 1

Patent Document 2

Summary of the Invention

Problems to be Solved by the Invention

[0015] An object of the present invention is to provide an amorphous novel compound or a salt thereof having excellent selective enhancing activity against melanocortin receptors, particularly melanocortin-4 receptor (MC4R).

[0016] Another object of the present invention is to provide a pharmaceutical composition comprising a stable crystalline form of the novel compound.

Means for Solving the Problems

[0017] To achieve the above object, In one aspect, the present invention provides a crystalline form IV of a compound of the following Chemical Formula 1, a pharmaceutically acceptable salt thereof, or a solvate thereof, which has, in an X-ray powder diffraction pattern, three or more, five or more, seven or more, nine or more, or ten or more characteristic peaks selected from the following diffraction angles (2θ values): 7.76 ± 0.2°, 9.69 ± 0.2°, 10.12 ± 0.2°, 10.63 ± 0.2°, 11.93 ± 0.2°, 12.72 ± 0.2°, 14.57 ± 0.2°, 16.07 ± 0.2°, 17.65 ± 0.2°, 18.24 ± 0.2°, 19.10 ± 0.2°, 19.62 ± 0.2°, 20.09 ± 0.2°, 21.19 ± 0.2°, 21.62 ± 0.2°, 22.02 ± 0.2°, 22.25 ± 0.2°, 22.61 ± 0.2°, 23.59 ± 0.2°, 24.26 ± 0.2°, 24.78 ± 0.2°, 25.29 ± 0.2°, 26.15 ± 0.2°, 28.52 ± 0.2°, 29.38 ± 0.2°, and 29.98 ± 0.2°.

[0018] [Chemical formula]

[0019] In Chemical Formula 1, R1 is C2-C5 alkyl.

[0020] The compound of Chemical Formula 1 can have an asymmetric carbon center and an asymmetric axis or plane, and can exist as cis or trans isomers, R or S isomers, racemates, mixtures of diastereoisomers, and individual diastereoisomers, and all these isomers and mixtures are included within the scope of the compound of Chemical Formula 1.

[0021] In this specification, for convenience, unless otherwise specified, the compound of Chemical Formula 1 is used in the sense of including all of the compound of Chemical Formula 1, its pharmaceutically acceptable salts, its isomers, and its solvates.

[0022] In one specific example according to the present invention, R1 of the Chemical Formula 1 is C2-C5 alkyl. In another specific example according to the present invention, R1 of the Chemical Formula 1 is linear or branched C2-C5 alkyl, for example, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, or tert-butyl.

[0023] In another specific example according to the present invention, R1 of the Chemical Formula 1 is C2 or C3 alkyl. In another specific example according to the present invention, R1 of the Chemical Formula 1 is linear or branched C2 or C3 alkyl, for example, ethyl, n-propyl, or iso-propyl.

[0024] In one specific example 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, and 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, and maleic acid, and sulfonic acids such as methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, or naphthalenesulfonic acid.

[0025] In one specific example according to the present invention, the solvate can include hydrates; solvates with organic solvents such as methanol, ethanol, 2-propanol, 1,2-propanediol, 1,3-propanediol, n-butanol, 1,4-butanediol, tert-butanol, acetic acid, acetone, butyl acetate, methyl acetate, ethyl acetate, propyl acetate, t-butyl acetate, isobutyl acetate, methyl ethyl ketone, 2-pentanone, tetrahydrofuran, acetonitrile, chloroform, toluene, and mixtures thereof.

[0026] In one specific example according to the present invention, the crystalline form IV can be a crystalline form of a pharmaceutically acceptable salt of the compound of Chemical Formula 1.

[0027] The pharmaceutically acceptable salt of the compound of Chemical Formula 1 can be the hydrochloride compound of the following Chemical Formula 2.

[0028]

Chem.

[0029] In Chemical Formula 2, R2 is C2-C5 alkyl.

[0030] In still another specific example according to the present invention, the pharmaceutically acceptable salt of the compound of Chemical Formula 1 can be 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)isobutylamide hydrochloride of the following Chemical Formula 3.

[0031]

Chem.

[0032] In another specific example according to the present invention, the crystalline form IV can be a solvate, specifically a hydrate crystalline form, of a pharmaceutically acceptable salt of the compound of Chemical Formula 1.

[0033] More specifically, the crystalline form IV can be a crystalline form of a hydrate of the hydrochloride salt of the compound of Chemical Formula 1.

[0034] In one specific example according to the present invention, the crystalline form IV can be a crystalline form of the compound of the following Chemical Formula 4.

[0035]

Chem.

[0036] Crystal form IV according to the present invention shows three or more, five or more, seven or more, nine or more, or ten or more characteristic peaks selected from 7.76 ± 0.2°, 9.69 ± 0.2°, 10.12 ± 0.2°, 10.63 ± 0.2°, 11.93 ± 0.2°, 12.72 ± 0.2°, 14.57 ± 0.2°, 16.07 ± 0.2°, 17.65 ± 0.2°, 18.24 ± 0.2°, 19.10 ± 0.2°, 19.62 ± 0.2°, 20.09 ± 0.2°, 21.19 ± 0.2°, 21.62 ± 0.2°, 22.02 ± 0.2°, 22.25 ± 0.2°, 22.61 ± 0.2°, 23.59 ± 0.2°, 24.26 ± 0.2°, 24.78 ± 0.2°, 25.29 ± 0.2°, 26.15 ± 0.2°, 28.52 ± 0.2°, 29.38 ± 0.2°, and 29.98 ± 0.2° during X-ray powder diffraction (XRD) analysis.

[0037] In one specific example according to the present invention, the crystal form IV can have the XRD pattern shown in FIG. 4.

[0038] Crystal form IV according to the present invention shows three endothermic peaks at 30 to 200°C and an endothermic peak due to decomposition at 220°C or higher in a differential scanning calorimetry (DSC) profile.

[0039] In one specific example according to the present invention, the crystal form IV can have the DSC profile shown in FIG. 5.

[0040] Crystal form IV according to the present invention can have a weight loss of 10% or less, for example, 1% to 10%, 3% to 8%, or 5% when heated at a temperature of 140°C or lower in a thermogravimetric analysis (TGA) profile.

[0041] In one specific example according to the present invention, the crystal form IV can have the TGA profile shown in FIG. 6.

[0042] In this specification, The results of X-ray diffraction (XRD) analysis were obtained using a PANalytical X’ Pert Pro MPD system, Malvern Panalytical Ltd.

[0043] The results of differential scanning calorimetry (DSC) analysis were obtained using a DSC1, Mettler-Toledo AG.

[0044] The results of thermogravimetric analysis (TGA) were obtained using a TGA / DSC 1, Mettler-Toledo AG.

[0045] The crystalline form IV can have a higher purity compared to the crude compound of formula 1, the amorphous compound of formula 1, or other crystalline forms of the compound of formula 1, and can be physically and chemically more stable.

[0046] Moreover, the crystalline form IV of the compound of formula 1 can be further superior in the enhancing ability for the melanocortin-4 receptor and the preventive or therapeutic effect on diseases such as obesity, diabetes, inflammation, erectile dysfunction, etc., compared to known melanocortin-4 receptor agonists, but the effects of the present invention are not limited thereto.

[0047] In another aspect, the present invention provides a method for producing crystalline form IV, comprising the steps of dissolving the compound of formula 1 in a crystallization solvent to produce a mixed solution, and obtaining crystals from the mixed solution.

[0048] First, the compound represented by formula 1 is dissolved in a crystallization solvent.

[0049] For the production of the crystalline form IV, the compound of formula 1 can be the compound of formula 1, its salt, its isomer, or a solvate thereof.

[0050] The compound of Chemical Formula 1 can be obtained by the production method described in the specification of Korean Application No. 10-2019-0141649 (filed on November 7, 2019).

[0051] The crystallization solvent can be used without particular limitation as long as it is an appropriate solvent for crystallization of the compound. In one specific example, the crystallization solvent includes a mixture of water and a polar aprotic organic solvent.

[0052] The polar aprotic organic solvent can include ethyl acetate, methyl isobutyl ketone, dimethylsulfoxide, tetrahydrofuran, acetone, dimethylformamide, acetonitrile, or a mixture thereof.

[0053] In one specific example according to the present invention, the polar aprotic organic solvent can include ethyl acetate.

[0054] In one specific example according to the present invention, the crystallization solvent can be a mixed solvent in which water and a polar aprotic organic solvent are mixed at a volume ratio of 35:1 to 1:35, specifically, 20:1 to 1:30, 10:1 to 1:25, 1:1 to 1:20, 1:10 to 1:30, 1:15 to 1:25, 1:17 to 1:23, 1:18 to 1:22, or 1:20.

[0055] With respect to 1 g of the compound of Chemical Formula 1, 0.5 to 5 mL, 0.7 to 3 mL, 0.8 to 2.5 mL, 1 to 2.4 mL, 1.5 to 2.3 mL, 2.0 to 2.2 mL, or 2.1 mL of the crystallization solvent can be used.

[0056] The dissolution of the compound of Chemical Formula 1 in a crystallization solvent can be carried out without stirring or with stirring at room temperature, for example, 20 to 30 °C, specifically, 23 to 28 °C or 25 °C.

[0057] In one specific example according to the present invention, for 1 g of the compound of Chemical Formula 1, 2 mL of EtOAc and 0.1 mL of distilled water can be used to obtain a mixed solution in which the compound of Chemical Formula 1 is dissolved at room temperature.

[0058] Next, it includes the step of obtaining crystals from the mixed solution in which the compound of Chemical Formula 1 is dissolved. The obtaining of the crystals can be carried out, for example, by using methods such as cooling the solution, dropping an acid into the solution to form a precipitate, evaporating the solvent, adding an antisolvent to supersaturate, or slurry conversion.

[0059] Also, the crystallization step can include stirring the mixed solution. The stirring can be carried out by known means, and the stirring time is not limited thereto, but for example, it can be carried out for 15 hours or more and 50 hours or less, specifically, 15 hours to 50 hours, 15 hours to 45 hours, 15 hours to 40 hours, 15 hours to 35 hours, 20 hours to 30 hours, 23 hours to 28 hours, 23 to 25 hours or 24 hours.

[0060] In another specific example according to the present invention, the precipitate formed by stirring the mixed solution at room temperature can be filtered and washed to obtain crystals.

[0061] In still other specific examples according to the present invention, before, after or simultaneously with the stirring of the mixed solution, the step of adding a non-polar organic solvent to the mixed solution can be further included. By adding the non-polar organic solvent to increase the generation rate of the crystallization particles, the yield or production stability of crystalline form IV can be made excellent, but the present invention is not limited thereto. The non-polar organic solvent can be used without particular limitation as long as it is an organic solvent having non-polar characteristics. For example, hexane, heptane, cyclohexane, carbon tetrachloride, benzene, chloroform, etc. can be used. In one specific example according to the present invention, the step of introducing heptane into the solution during crystallization from the mixed solution can be included.

[0062] The crystalline form IV obtained as described above can have a higher purity and can be physically and chemically more stable than the crude compound of Chemical Formula 1, the amorphous compound of Chemical Formula 1 or any crystalline form of Chemical Formula 1, but the effects of the present invention are not limited thereto.

[0063] In still other aspects, the present invention provides a pharmaceutical composition comprising (i) the crystalline form IV; and (ii) a pharmaceutically acceptable carrier.

[0064] Since the crystalline form IV according to the present invention exhibits an excellent enhancing effect on melanocortin receptors, particularly melanocortin-4 receptor (MC4R), the present invention can also provide a pharmaceutical composition for enhancing the function of melanocortin receptors containing the above-described crystalline form IV as an active ingredient. Specifically, the pharmaceutical composition can be a composition for enhancing the function of melanocortin-4 receptor.

[0065] In addition, the pharmaceutical composition can exhibit excellent effects on the prevention or treatment of obesity, diabetes, inflammation and erectile dysfunction, and can be a composition for the prevention or treatment of obesity, the prevention or treatment of diabetes, the prevention or treatment of inflammation, or the prevention or treatment of erectile dysfunction, but the uses of the present invention are not limited only to these diseases.

[0066] As used herein, "carrier" means a compound that facilitates the introduction of a compound into a cell or tissue.

[0067] When the crystalline form IV of the present invention is administered for clinical purposes, the total daily dose administered to the host in a single volume or divided volumes is preferably in the range of 0.01 to 10 mg per kg of body weight, but the specific dose level for an individual patient can vary depending on the specific compound being used, the patient's body weight, gender, health status, diet, time of administration of the drug, method of administration, excretion rate, drug combination, and severity of the disease, among other factors.

[0068] The crystalline form IV of the present invention can be administered by any route depending on the purpose. For example, the amorphous compound of the present invention can be administered by injection or orally.

[0069] The pharmaceutical composition of the present invention can be in various oral dosage forms such as tablets, pills, powders, capsules, granules, syrups or emulsions, or in parenteral dosage forms such as injectable preparations for intramuscular, intravenous or subcutaneous administration.

[0070] Injectable preparations can be manufactured using suitable dispersing agents, wetting agents, suspending agents, or excipients according to known techniques.

[0071] Excipients that can be used in the pharmaceutical preparations of the present invention include, but are not limited to, sweeteners, binders, solubilizers, solubilizing aids, wetting agents, emulsifiers, isotonic agents, adsorbents, disintegrants, antioxidants, preservatives, lubricants, fillers, fragrances, etc. For example, as excipients, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, silica, magnesium aluminum silicate, starch, gelatin, tragacanth gum, alginic acid, sodium alginate, methyl cellulose, sodium carboxymethyl cellulose, water, ethanol, polyethylene glycol, polyvinyl pyrrolidone, sodium chloride, calcium chloride, orange essence, strawberry essence, vanilla fragrance, etc. can be used.

[0072] When the pharmaceutical composition of the present invention is in the form of an oral administration, examples of carriers used include, but are not limited to, cellulose, calcium silicate, corn starch, lactose, sucrose, dextrose, calcium phosphate, stearic acid, magnesium stearate, calcium stearate, gelatin, talc, etc.

[0073] When the pharmaceutical composition of the present invention is in the form of an injectable preparation, examples of the carrier include, but are not limited to, water, saline, glucose aqueous solution, pseudo-sugar aqueous solution, alcohol, glycol, ether, oil, fatty acid, fatty acid ester, glyceride, etc.

[0074] In yet another aspect, provided is the above-mentioned crystalline form IV for use in enhancing the function of melanocortin receptors, particularly melanocortin-4 receptor (MC4R).

[0075] In one embodiment, provided is the above-mentioned crystalline form IV for use in the treatment or prevention of obesity, diabetes, inflammation or erectile dysfunction.

[0076] In yet another aspect, there is provided a method for enhancing the function of a melanocortin receptor, particularly the melanocortin-4 receptor (MC4R), which comprises the step of administering the above-described crystalline form IV to a subject.

[0077] In yet another aspect, there is provided a method for treating obesity, diabetes, inflammation or erectile dysfunction, which comprises the step of administering the above-described crystalline form IV to a subject.

Advantages of the Invention

[0078] The crystalline form IV according to the present invention exhibits an excellent enhancing effect on melanocortin receptors, particularly the melanocortin-4 receptor (MC4R), and thus can be usefully used for the prevention or treatment of obesity, diabetes, inflammation and erectile dysfunction.

[0079] The crystalline form IV according to the present invention shows an on-target effect on the melanocortin-4 receptor, exhibits weight loss and diet reduction effects, does not affect anxiety and depression, and can be administered without side effects such as hERG (human ether-a-go-go related gene) inhibition and stability problems such as induction of mutations.

[0080] In addition, the crystalline form IV according to the present invention is superior in purity, yield, physical and chemical stability compared to the crude compound of Chemical Formula 1, the amorphous compound of Chemical Formula 1 or any other crystalline form of Chemical Formula 1.

[0081] Specifically, the crystalline form IV can be superior in solubility, storage stability and production stability compared to the crude compound of Chemical Formula 1, the amorphous compound of Chemical Formula 1 or any other crystalline form of Chemical Formula 1.

Brief Description of the Drawings

[0082]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Mode for Carrying Out the Invention

[0083] Hereinafter, the present invention will be described more specifically with reference to Production Examples and Examples. However, these Examples are illustrative of the present invention, and the scope of the present invention is not limited thereto.

[0084] Production Example 1: Production of Methyl (2S,4S)-4-(N-((1s,4R)-4-methylcyclohexyl)isobutylamide)pyrrolidine-2-carboxylate Hydrochloride

Chemical formula

[0085] The title compound was obtained through the processes of the following Steps A, B, C, D, and E.

[0086] Step A: Preparation of 1-(tert-butyl) 2-methyl (2S,4S)-4-azidopyrrolidine-1,2-dicarboxylate Under nitrogen, 1-(tert-butyl) 2-methyl (2S,4R)-4-((methylsulfonyl)oxy)pyrrolidine-1,2-dicarboxylate (48.5 g, 150 mmol) was dissolved in N,N'-dimethylformamide (250 ml), and sodium azide (19.5 g, 300 ml) was added. Stirring was carried out at 80 °C for 16 hours. After concentrating the reaction solvent under reduced pressure, water was added, and extraction was performed twice with ethyl acetate. The organic layer was washed with an aqueous sodium chloride solution and water, and then dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain crude 1-(tert-butyl) 2-methyl (2S,4S)-4-azidopyrrolidine-1,2-dicarboxylate (39.59 g, 98%), which was used in the next step without purification.

[0087] 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)

[0088] Step B: Preparation of 1-(tert-butyl) 2-methyl (2S,4S)-4-aminopyrrolidine-1,2-dicarboxylate 1-(tert-Butyl) 2-methyl (2S,4S)-4-azidopyrrolidine-1,2-dicarboxylate (24.59 g, 91.0 mmol) obtained in the above step A was dissolved in tetrahydrofuran (180 ml), and then 1M trimethylphosphine tetrahydro solution (109.2 ml, 109.2 mmol) was gradually added at 0 °C. After stirring at the same temperature for 1 hour, the mixture was stirred at room temperature for 3 hours. After concentrating the reaction solvent under reduced pressure, dichloromethane (100 ml) and water (150 ml) were added and stirred for about 30 minutes. After liquid separation and extraction with dichloromethane again, the organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain crude 1-(tert-butyl) 2-methyl (2S,4S)-4-aminopyrrolidine-1,2-dicarboxylate (20.62 g, 93%), which was used in the next step without purification.

[0089] 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)

[0090] Step C: Preparation of 1-(tert-butyl) 2-methyl (2S,4S)-4-(((1s,4R)-4-methylcyclohexyl)amino)pyrrolidine-1,2-dicarboxylate The 1-(tert-butyl) 2-methyl (2S,4S)-4-aminopyrrolidine-1,2-dicarboxylate (20.62 g, 84.4 mmol) obtained in the previous step B was dissolved in dichloroethane (150 ml), and 4-methylcyclohexanone (9.5 ml, 101.3 mmol) was added. Sodium triacetoxyborohydride (26.8 g, 126.6 mmol) was added at 0 °C, and the mixture was stirred at room temperature for 16 hours. The reaction solvent was concentrated under reduced pressure, water was added, and the mixture was extracted twice with ethyl acetate. The organic layer was washed with an aqueous sodium chloride solution and then dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure and purified by column chromatography to obtain 1-(tert-butyl) 2-methyl (2S,4S)-4-(((1s,4R)-4-methylcyclohexyl)amino)pyrrolidine-1,2-dicarboxylate (22.9 g, 80%).

[0091] 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)

[0092] Step D: Preparation of 1-(tert-butyl) 2-methyl (2S,4S)-4-(N-((1s,4R)-4-methylcyclohexyl)isobutylamide)pyrrolidine-1,2-dicarboxylate 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 dichloromethane (500 ml), triethylamine (61.1 ml, 438.1 mmol) was added, and then isobutyryl chloride (11.7 ml, 219 mmol) was gradually added at 0 °C. After stirring at room temperature for 16 hours, the reaction solvent was concentrated under reduced pressure, an aqueous sodium hydrogen carbonate solution was added, and extraction was performed twice 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 and purified by column chromatography to obtain 1-(tert-butyl) 2-methyl (2S,4S)-4-(N-((1s,4R)-4-methylcyclohexyl)isobutyramido)pyrrolidine-1,2-dicarboxylate (38.79 g, 86%).

[0093] 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)

[0094] Step E: Preparation of methyl (2S,4S)-4-(N-((1s,4R)-4-methylcyclohexyl)isobutylamide)pyrrolidine-2-carboxylate hydrochloride 1-(tert-Butyl) 2-methyl (2S,4S)-4-(N-((1s,4R)-4-methylcyclohexyl)isobutylamide) pyrrolidine-1,2-dicarboxylate (34.0 g, 82.8 mmol) obtained in the above step D was dissolved in dichloromethane (200 ml), and then 4N hydrochloric acid 1,4-dioxane solution (82.8 ml, 331.3 mmol) was added at 0 °C. After stirring at room temperature for 6 hours, the reaction solvent was concentrated under reduced pressure to obtain crude (28.7 g, 99%), which was used in the next step without purification.

[0095] MS[M+H] = 311 (M+1)

[0096] Production Example 2: Preparation of (3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carboxylic acid

Chemical formula

[0097] The title compound was obtained by the method described in International Publication No. WO2004 / 092126.

[0098] 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 (dd, J = 11.2, 10.0 Hz, 1H), 3.29 (dd, J = 11.2, 11.2 Hz, 1H), 3.18-3.09 (m, 1H), 1.44 (s, 9H)

[0099] Production Example 3: 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)isobutylamide

Chemical formula

[0100] The title compound was obtained through the processes of the following steps A, B, and C.

[0101] 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)isobutylamide)pyrrolidine-2-carboxylate Methyl (2S,4S)-4-(N-((1s,4R)-4-methylcyclohexyl)isobutylamide)pyrrolidine-2-carboxylate hydrochloride (28.7 g, 82.73 mmol) obtained in Production Example 1, (3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carboxylic acid (24.5 g, 86.87 mmol) obtained in Production Example 2, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (22.2 g, 115.83 mmol), and 1-hydroxybenzotriazole hydrate (15.7 g, 115.83 mmol) were dissolved in N,N'-dimethylformamide (400 ml), and N,N'-diisopropylethylamine (72.0 ml, 413.66 mmol) was gradually added. The mixture was stirred at room temperature for 16 hours. After concentrating the reaction solvent under reduced pressure, 0.5 N aqueous sodium hydroxide solution was added, and the mixture was extracted twice with ethyl acetate. The organic layer was washed twice each with 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 methyl (2S,4S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-4-(N-((1s,4R)-4-methylcyclohexyl)isobutylamide)pyrrolidine-2-carboxylate (41.19 g, 87%).

[0102] MS [M+H] = 575 (M+1)

[0103] Step B: Preparation of (2S,4S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-4-(N-((1s,4R)-4-methylcyclohexyl)isobutylamide)pyrrolidine-2-carboxylic acid Methyl (2S,4S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-4-(N-((1s,4R)-4-methylcyclohexyl)isobutylamide)pyrrolidine-2-carboxylate (39.4 g, 68.62 mmol) obtained in the previous step A was dissolved in methanol (450 ml), and then 6N aqueous sodium hydroxide solution (57.2 ml, 343.09 mmol) was added. The mixture was stirred at room temperature for 16 hours, adjusted to about pH 5 with 6N aqueous hydrochloric acid solution, and then the reaction solution was concentrated under reduced pressure. The concentrated solution was dissolved in dichloromethane, and the undissolved solid was filtered through a paper filter. The filtrate was concentrated under reduced pressure to obtain the crude title compound (38.4 g, 99%), which was used in the next step without purification.

[0104] MS [M+H] = 561 (M+1)

[0105] 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)isobutylamide (2S,4S)-1-((3S,4R)-1-(tert-Butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-4-(N-((1s,4R)-4-methylcyclohexyl)isobutylamide)pyrrolidine-2-carboxylic acid (38.4 g, 68.60 mmol), 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 gradually added in turn. The mixture was stirred at room temperature for 16 hours, and the reaction solution was concentrated under reduced pressure. Then, 0.5 N 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 dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure and purified by column chromatography to obtain N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidine-3-yl)-N-((1s,4R)-4-methylcyclohexyl)isobutylamide (37.05 g, 86%).

[0106] MS [M+H] = 630 (M+1)

[0107] Production Example 4: Preparation of amorphous compound of N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidine-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidine-3-yl)-N-((1s,4R)-4-methylcyclohexyl)isobutylamide hydrochloride

Chemical Structure

[0108] Based on 1 g of the compound (MC70) produced in Production Example 3, 19 mL of MBTE was used to dissolve the compound (MC70) at 25°C. After complete dissolution, 1 mL of heptane was added, and then the mixture was cooled to -5 to 0°C. After reaching the set temperature, 1 equivalent of 4M HCl / EtOAc was added dropwise, followed by stirring for about 90 minutes and filtration to obtain the title compound (MC71). (Yield: about 90%)

[0109] The XRD (Figure 1), DSC (Figure 2), and TGA (Figure 3) analysis results for the compound of Production Example 4 are attached to Figures 1 to 3 respectively, and it was confirmed from the analysis results that the compound is an amorphous compound. The analysis methods for XRD, DSC, and TGA are as described later in the experimental example for Example 1 below.

[0110] Example 1. Production of Crystal Form IV of Hydrate of N-((3S,5S)-1-((3S,4R)-1-(tert-butyl)-4-(4-chlorophenyl)pyrrolidin-3-carbonyl)-5-(morpholine-4-carbonyl)pyrrolidin-3-yl)-N-((1s,4R)-4-methylcyclohexyl)isobutylamide Hydrochloride

Chemical Structure

[0111] 1 g of the compound (MC71) of Production Example 4 produced above was dissolved at room temperature using 2 mL of EtOAc and 0.1 mL of distilled water. After complete dissolution, the mixture was stirred at room temperature for 24 hours using an electromagnetic stirrer, and then filtered under nitrogen pressure to obtain the title Crystal Form IV. (Yield: about 72%)

[0112] Comparative Example 1 1 g of the compound (MC71) of Production Example 4 produced above was dissolved at room temperature using 1 mL of EtOAc. After complete dissolution, the mixture was stirred with an electromagnetic stirrer for about 43 hours, but no crystals like those in Example 1 were obtained.

[0113] Experimental Example 1. XRD Evaluation The powder XRD diffraction pattern was obtained by the following method using a PANalytical X’Pert Pro MPD system equipped with a monochromatized radiation source and a Ni filter as a solid-state detector.

[0114] After about 20 - 30 mg of the sample was evenly placed on a glass sample holder so as to have a flat surface, the generator of the instrument was set to 45 kV (acceleration voltage) and 40 mA (filament emission), and then the measurement was carried out in reflection mode (not-spin). The Bragg angle (2θ) in the range of 4 - 40° was measured under the conditions of a step size of 0.026° and a time per step of 51 seconds.

[0115] The result of measuring the obtained crystalline form IV by XRD is shown in Figure 4.

[0116] As confirmed from the spectrum shown in Figure 4, the crystalline form IV according to the present invention shows characteristic peaks (2θ) at 7.76°, 9.69°, 10.12°, 10.63°, 11.93°, 12.72°, 14.57°, 16.07°, 17.65°, 18.24°, 19.10°, 19.62°, 20.09°, 21.19°, 21.62°, 22.02°, 22.25°, 22.61°, 23.59°, 24.26°, 24.78°, 25.29°, 26.15°, 28.52°, 29.38° and 29.98°. The specific values of XRD are shown in Table 1 below.

[0117]

Table 1

[0118] Experimental Example 2. Differential Scanning Calorimetry (DSC) DSC was measured using a Mettler Toledo DSC1 system. A sample of about 2 - 5 mg was weighed and placed in a 40 μL Al crucible (flat-bottomed aluminum pan with one pin-hole lid) to form one pin hole. Next, the sample was heated from 25 °C to 350 °C at a rate of 10 °C / min to measure DSC. During the measurement, nitrogen gas was supplied into the apparatus at a rate of 70 mL / min to prevent the inflow of oxygen and other gases. Data collection and evaluation were performed using the software STARe.

[0119] The results of DSC measurement of the obtained crystalline form IV are shown in Figure 5.

[0120] As confirmed from Figure 5, a total of three endothermic peaks were observed for crystalline form IV at approximately 63.6 °C (Onset), 133.4 °C (Onset), and 178.2 °C (Onset). After approximately 220 °C, an endothermic peak due to decomposition was shown. The temperature values have an error of ±5 °C.

[0121] Experimental Example 3. Thermogravimetric Analysis (TGA) TGA was measured using a Mettler Toledo TGA / DSC 1 module. A sample of about 4 - 8 mg was weighed and placed in a 100 μL Al crucible (flat-bottomed aluminum crucibles). Next, the sample was heated from 30 °C to 350 °C at a rate of 10 °C / min to measure TGA. During the measurement, nitrogen gas was supplied into the apparatus at a rate of 80 mL / min to prevent the inflow of oxygen and other gases. Data collection and evaluation were performed using the software STARe.

[0122] The results of TGA measurement of the obtained crystalline form IV are shown in Figure 6.

[0123] As confirmed from Figure 6, for crystalline form IV, a weight loss of approximately 5.0% was observed at a temperature below 100°C. After approximately 220°C, a weight loss due to decomposition occurred. The temperature values have an error of ±5°C.

Claims

1. A crystalline form IV of a compound of the following formula 1, a pharma- ceutically acceptable salt thereof, or a solvate thereof: The X-ray powder diffraction pattern showed the following diffraction angles (2θ values): 7.76±0.2°, 9.69±0.2°, 10.12±0.2°, 10.63±0.2°, 11.93±0.2°, 12.72±0.2°, 14.57±0.2°, 16.07±0.2°, 17.65±0.2°, 18.24±0.2°, 19.10±0.2°, 19.62±0.2°, 20.09±0.2°, 21.1 9±0.2°, 21.62±0.2°, 22.02±0.2°, 22.25±0.2°, 22.61±0.2°, 23.59±0.2°, 24.26±0.2°, 24.78±0.2°, 25.29±0.2°, 26.15±0.2°, 28.52±0.2°, 29.38±0.2°, and 29.98±0.2°. 【Chemistry 1】 In the above Chemical Formula 1, R １ is C ２ -C ５ It is an alkyl.

2. 2. The crystalline form IV of claim 1, wherein the pharma- ceutically acceptable salt of the compound of formula 1 is selected from the group consisting of hydrochloride, sulfate, nitrate, phosphate, hydrobromide and hydroiodide salts of the compound.

3. 2. The crystalline form IV of claim 1, which is a crystalline form of a solvate of the hydrochloride salt of the compound of formula 1.

4. The crystalline form IV of claim 3, wherein the solvate is a hydrate.

5. The crystalline form IV of the compound of formula 4 according to claim 4: 【Chemistry 2】 。

6. A method for preparing crystalline form IV according to any one of claims 1 to 5, comprising the steps of: dissolving the compound of Formula 1 in a crystallization solvent to prepare a mixed solution; and obtaining crystals from the mixed solution.

7. 7. The method for preparing crystalline form IV according to claim 6, wherein the crystallization solvent comprises water, a polar aprotic organic solvent or a mixture thereof.

8. The method for preparing the crystalline form IV according to claim 7, wherein the polar aprotic organic solvent comprises ethyl acetate, methyl isobutyl ketone, dimethylsulfoxide, tetrahydrofuran, acetone, dimethylformamide, acetonitrile or a mixture thereof.

9. The method for preparing the crystalline form IV according to claim 7, wherein the crystallization solvent is a mixed solvent of water and a polar aprotic organic solvent in a volume ratio of 35:1 to 1:

35.

10. 7. The method for producing the crystalline form IV of claim 6, wherein the crystallization step comprises stirring the mixed solution.

11. A pharmaceutical composition comprising the crystalline form IV of any one of claims 1 to 5 and a pharma- ceutically acceptable carrier.

12. A pharmaceutical composition for enhancing the activity of melanocortin-4 receptor, comprising the crystalline form IV of any one of claims 1 to 5 and a pharma- ceutically acceptable carrier.

13. The pharmaceutical composition of claim 12, wherein the composition is for the prevention or treatment of obesity, diabetes, inflammation or erectile dysfunction.

Images