Cyclic amine derivatives as adbilin function enhancers, novel cyclic amine derivatives, and their medicinal uses

Abstract

The present invention aims to provide an adbilin function promoter composed of a low molecular weight compound that is useful for treating nerve axonal damage. The present invention provides an adbilin function promoter containing a cyclic amine derivative represented by the following chemical formula or a pharmacologically acceptable salt thereof as an active ingredient.

Description

Technology Field

[0001] The present invention relates to cyclic amine derivatives as adbilin function promoters, novel cyclic amine derivatives, and their medicinal uses. Background Technology

[0002] Advilin is primarily expressed in peripheral nerves and plays an important role in nerve axon extension (Non-patent Literature 1 and Non-patent Literature 2). In mice deficient in Advilin, a failure in nerve axon extension during neural circuit regeneration is observed, and the length of the nerve axon is also short (Non-patent Literature 2). Furthermore, when nerve axon damage is induced in Advilin-deficient mice by nerve ligation or anticancer treatment, worsening of symptoms related to nerve axon damage is observed compared to normal mice (Non-patent Literature 2 and Non-patent Literature 3). On the other hand, in nerve cells that overexpress Advilin, the number and length of nerve processes containing nerve axons increase (Non-patent Literature 3). That is, it is known that Advilin has a function of promoting nerve axon extension during nerve regeneration.

[0003] In order for a nerve axon to extend, the polymerization of monomeric actin and the depolymerization of actin filaments (hereinafter, actin filament metabolic turnover) in the growth cone at the tip of the nerve axon need to be regulated normally (Non-patent Literature 4). It is known that the above-mentioned Advilin is present in the growth cone at the tip of the nerve axon (Non-patent Literature 3). Furthermore, Advilin is known to be an actin-binding protein that has the function of regulating actin filament metabolic turnover (Non-patent Literature 5).

[0004] Axonal damage to peripheral nerves is also referred to as peripheral nerve injury, but it is caused by surgical procedures, toxins, circulatory disorders, trauma resulting from traffic accidents, or radiation therapy, leading to motor paralysis, sensory paralysis, or autonomic nerve dysfunction. Although it is known that axonal regeneration occurs after peripheral nerve axonal damage, it often results in a failure to recover function, including abnormalities. It is said that one of the causes is that actual axonal regeneration requires time, and during this period, the tissues surrounding the axon or the target tissues junctioning with the axon terminals degenerate, preventing accurate rejunction between the axon terminals and the target tissues or target cells, thereby preventing effective and normal nerve innervation. Inaccuracies in rejunction occur not only between nerves and target tissues such as muscle tissue, but also between sensory and motor nerves; abnormal sensations have been observed between sensory nerves, and involuntary abnormal movements between motor nerves (Non-patent Literature 6). In addition, in particular, natural healing cannot be expected in cases of nerve axonal and nerve sheathing, and there are no effective treatments available, so surgical procedures such as nerve repair or functional reconstruction surgery are performed, but the problem is that the burden on the patient is heavy (Non-patent Literature 7). Therefore, there is a desire for an effective treatment that promotes nerve axonal extension to treat and prevent functional failure, including abnormalities following peripheral nerve axonal damage.

[0005] Patent documents 1 and 2 disclose that cyclic amine derivatives have analgesic effects and can treat or prevent peripheral nerve damage, but they do not disclose effects that promote adbilin function or treat nerve axonal damage. Furthermore, no compounds that promote adbilin function are known. Prior art literature

[0006] International Publication No. 2016 / 136944 International Publication No. 2018 / 181860

[0007] Ravenall et al., European Journal of Neuroscience, 2002, Vol. 15, pp. 281-290 Hasegawa et al., The Journal of Neuroscience, 2007, Vol. 27, pp. 14404-14414 Chuang et al., Proceedings of the National Academy of Sciences of the United States of America, 2018, Vol. 115, pp. E8557-E8566 Blanquie et al., Current Opinion in Neurobiology, 2018, Vol. 51, pp. 60-69 Rao et al., The Journal of Clinical Investigation, 2017, Vol. 127, pp. 4257-4269 Nishiwaki et al., Rehabilitation Medicine, 2002, Vol. 39, pp. 257-266 Kanaya, The Japanese Journal of Rehabilitation Medicine, 2014, Vol. 51, pp. 52-60 The problem to be solved

[0008] Therefore, the present invention aims to provide an adbilin function promoter composed of a low-molecular-weight compound that is useful for the treatment of nerve axonal damage. means of solving the problem

[0009] As a result of repeated research to solve the above problem, the inventors came to discover that a cyclic amine derivative or a pharmacologically acceptable salt thereof has an effect of promoting adbilin function.

[0010] That is, the present invention provides an adbilin function promoter containing a cyclic amine derivative represented by the following general formula (I) or a pharmacologically acceptable salt thereof as an active ingredient.

[0011]

[0012] [In the formula, carbons marked with * are non-predicated carbons, and A represents a group represented by the following general formula (IIa) or (IIb), and

[0013]

[0014] R 1 represents a methyl group, n-propyl group, isopropyl group, 2-methoxyethyl group, 2-ethoxyethyl group, difluoromethyl group, or 3,3,3-trifluoropropyl group, and R 2 represents a hydrogen atom, fluorine atom, or chlorine atom, and R 3 Each independently represents a methyl or ethyl group, and X is -O- or -N(R 3 )- represents]

[0015] In the above cyclic amine derivative, it is preferable that A is a group represented by the general formula (IIa), and then R 1 It is more preferable that the silver be a methyl group, an n-propyl group, an isopropyl group, a 2-methoxyethyl group, or a 3,3,3-trifluoropropyl group.

[0016] In the above-described cyclic amine derivative, it is preferable that A is a group represented by the general formula (IIb), and then X is -N(R 3 It is more desirable that )-is, and R 1 It is more preferable that the silver be a methyl group, an n-propyl group, an isopropyl group, a 2-methoxyethyl group, or a 3,3,3-trifluoropropyl group.

[0017] In addition, in the above-mentioned cyclic amine derivative, R 2 It is more preferable that it be a hydrogen atom or a chlorine atom.

[0018] In addition, in the above-mentioned cyclic amine derivative, it is more preferable that the stereochemistry of the sub-carbon with * attached is the S configuration.

[0019] By limiting it to these, the function-promoting effect of Advilin can be further enhanced.

[0020] The present invention provides an adbilin function promoter comprising, as an active ingredient, a cyclic amine derivative represented by the general formula (I) or a pharmacologically acceptable salt thereof, for promoting the function of adbilin by binding to adbilin and / or an adbilin complex.

[0021] In addition, the present invention provides an adbilin function promoter containing a cyclic amine derivative represented by the general formula (I) or a pharmacologically acceptable salt thereof as an active ingredient for improving abnormal regulation of actin filament metabolic turnover.

[0022] In addition, the present invention provides an adbilin function promoter containing a cyclic amine derivative represented by the general formula (I) or a pharmacologically acceptable salt thereof as an active ingredient for promoting nerve axon extension.

[0023] In addition, the present invention provides an adbilin function promoter comprising, as an active ingredient, a cyclic amine derivative represented by the general formula (I) or a pharmacologically acceptable salt thereof, for binding to adbilin and / or an adbilin complex to improve abnormal regulation of actin filament metabolic turnover and to promote nerve axon extension.

[0024] In addition, the present invention provides an adbilin function promoter containing, as an active ingredient, a cyclic amine derivative represented by the general formula (I) or a pharmacologically acceptable salt thereof, which is a treatment for nerve axonal damage.

[0025] In addition, the present invention relates to 3-hydroxy-3-(1-methyl-1H-imidazole-2-yl)-1-(4-morpholinopiperidin-1-yl)propan-1-one, 1-(4-(dimethylamino)piperidin-1-yl)-3-(1-propyl-1H-imidazole-2-yl)-3-hydroxypropan-1-one, 1-(4-(dimethylamino)piperidin-1-yl)-3-(1-isopropyl-1H-imidazole-2-yl)-3-hydroxypropan-1-one, 3-(5-chloro-1-methyl-1H-imidazole-2-yl)-1-(4-(dimethylamino)piperidin-1-yl)-3-hydroxypropan-1-one, Provides a cyclic amine derivative or a pharmacologically acceptable salt thereof, which is a compound selected from the group consisting of 1-(4-(dimethylamino)piperidin-1-yl)-3-(1-(2-methoxyethyl)-1H-imidazole-2-yl)-3-hydroxypropan-1-one and 1-(4-(dimethylamino)piperidin-1-yl)-3-(1-(3,3,3-trifluoropropyl)-1H-imidazole-2-yl)-3-hydroxypropan-1-one.

[0026] In addition, the present invention relates to 3-hydroxy-3-(1-methyl-1H-imidazole-2-yl)-1-(4-morpholinopiperidin-1-yl)propan-1-one, 1-(4-(dimethylamino)piperidin-1-yl)-3-(1-propyl-1H-imidazole-2-yl)-3-hydroxypropan-1-one, 1-(4-(dimethylamino)piperidin-1-yl)-3-(1-isopropyl-1H-imidazole-2-yl)-3-hydroxypropan-1-one, 3-(5-chloro-1-methyl-1H-imidazole-2-yl)-1-(4-(dimethylamino)piperidin-1-yl)-3-hydroxypropan-1-one, The present invention provides a medicine containing, as an active ingredient, a cyclic amine derivative or a pharmacologically acceptable salt thereof which is a compound selected from the group consisting of 1-(4-(dimethylamino)piperidin-1-yl)-3-(1-(2-methoxyethyl)-1H-imidazole-2-yl)-3-hydroxypropan-1-one and 1-(4-(dimethylamino)piperidin-1-yl)-3-(1-(3,3,3-trifluoropropyl)-1H-imidazole-2-yl)-3-hydroxypropan-1-one.

[0027] In addition, the present invention provides a pharmaceutical composition for treating nerve axonal damage, comprising a cyclic amine derivative represented by the above general formula (I), a pharmacologically acceptable salt thereof, and pharmacologically acceptable excipients, etc.

[0028] In addition, the present invention provides a cyclic amine derivative represented by the above general formula (I) or a pharmacologically acceptable salt thereof for use in the treatment of nerve axonal damage.

[0029] In addition, the present invention provides the use of a cyclic amine derivative represented by the above general formula (I) or a pharmacologically acceptable salt thereof for treating nerve axonal damage.

[0030] In addition, the present invention provides the use of a cyclic amine derivative represented by the above general formula (I) or a pharmacologically acceptable salt thereof in the manufacture of a medicine for treating nerve axonal damage.

[0031] In addition, the present invention provides a method for treating nerve axonal damage comprising administering a therapeutically effective amount of a cyclic amine derivative represented by the above general formula (I) or a pharmacologically acceptable salt thereof to a patient requiring treatment.

[0032] In addition, the present invention provides a method for treating nerve axonal damage comprising contacting a nerve cell with an effective amount of a cyclic amine derivative represented by the above general formula (I) or a pharmacologically acceptable salt thereof.

[0033] In addition, the present invention provides a method for treating nerve axonal damage comprising administering an effective amount of a cyclic amine derivative represented by the above general formula (I) or a pharmacologically acceptable salt thereof to a subject in need of it.

[0034] In addition, the causes of nerve axonal damage mentioned above are not limited to these, but include surgical procedures, toxins, circulatory disorders, trauma resulting from traffic accidents, or radiation therapy.

[0035] In addition, the present invention provides a pharmaceutical composition for treating a disease related to nerve axonal damage, comprising a cyclic amine derivative represented by the above general formula (I), a pharmacologically acceptable salt thereof, and a pharmacologically acceptable excipient.

[0036] In addition, the present invention provides a cyclic amine derivative represented by the above general formula (I) or a pharmacologically acceptable salt thereof for use in the treatment of diseases related to nerve axonal damage.

[0037] In addition, the present invention provides the use of a cyclic amine derivative represented by the above general formula (I) or a pharmacologically acceptable salt thereof for treating diseases related to nerve axonal damage.

[0038] In addition, the present invention provides the use of a cyclic amine derivative represented by the above general formula (I) or a pharmacologically acceptable salt thereof in the manufacture of a medicine for treating a disease related to nerve axonal damage.

[0039] In addition, the present invention provides a method for treating a disease related to nerve axonal damage, comprising administering a therapeutically effective amount of a cyclic amine derivative represented by the above general formula (I) or a pharmacologically acceptable salt thereof to a patient requiring treatment.

[0040] In addition, the present invention provides a method for treating a disease related to nerve axon damage, comprising contacting a nerve cell with an effective amount of a cyclic amine derivative represented by the above general formula (I) or a pharmacologically acceptable salt thereof.

[0041] In addition, the present invention provides a method for treating a disease related to nerve axonal damage, comprising administering an effective amount of a cyclic amine derivative represented by the above general formula (I) or a pharmacologically acceptable salt thereof to a subject who needs it.

[0042] In addition, the present invention provides a pharmaceutical composition for promoting adbilin function, for improving abnormal regulation of actin filament metabolic turnover, or for promoting nerve axon extension, containing a cyclic amine derivative represented by the above general formula (I), a pharmacologically acceptable salt thereof, and pharmacologically acceptable excipients, etc.

[0043] In addition, the present invention provides a cyclic amine derivative represented by the above general formula (I) or a pharmacologically acceptable salt thereof for use in promoting adbilin function, improving the regulation of actin filament metabolic turnover, or promoting nerve axon extension.

[0044] In addition, the present invention provides for the use of a cyclic amine derivative represented by the general formula (I) above or a pharmacologically acceptable salt thereof to promote adbilin function, to improve abnormal regulation of actin filament metabolic turnover, or to promote nerve axon extension.

[0045] In addition, the present invention provides the use of a cyclic amine derivative represented by the above general formula (I) or a pharmacologically acceptable salt thereof in the manufacture of a medicine for promoting adbilin function, for improving abnormalities in actin filament metabolic rotation regulation, or for promoting nerve axon extension.

[0046] In addition, the present invention provides a method for promoting adbilin function, improving abnormalities in actin filament metabolic rotation regulation, or promoting nerve axon extension, comprising administering a therapeutically effective amount of a cyclic amine derivative represented by the above general formula (I) or a pharmacologically acceptable salt thereof to a patient who requires this.

[0047] In addition, the present invention provides a method for promoting adbilin function, improving abnormalities in actin filament metabolic rotation regulation, or promoting nerve axon extension, comprising contacting a nerve cell with an effective amount of a cyclic amine derivative represented by the above general formula (I) or a pharmacologically acceptable salt thereof.

[0048] In addition, the present invention provides a method for promoting adbilin function, improving abnormalities in actin filament metabolic rotation regulation, or promoting nerve axon extension, comprising administering an effective amount of a cyclic amine derivative represented by the above general formula (I) or a pharmacologically acceptable salt thereof to a subject who requires it. Effects of the invention

[0049] According to the cyclic amine derivative of the present invention or its pharmacologically acceptable salt, the function of adbilin, one of the molecules regulating actin filament metabolic turnover, can be promoted and thus can be used as a medicine for diseases related to nerve axonal damage.

[0050] This specification includes the contents described in the specification and / or drawings of Japanese Patent Application No. 2018-243042, which forms the basis of the priority of this application. Brief explanation of the drawing

[0051] Figure 1 is a diagram showing the function of adbilin on actin filaments. Figure 2 is a diagram showing the specific binding of the compound of Example 7 to the membrane fraction prepared from each tissue of a rat. Figure 3 is a diagram showing the improved effect of the compound of Example 7 on the regulation of actin filament metabolic rotation in a rat spinal cord ligation model. Figure 4 is an image showing the neuronal axon stretching action of the compound of Example 7 in rat dorsal root ganglion primary culture cells that induced neuronal axonal damage as a change in cell morphology. Figure 5 is a diagram showing the neuronal axon extension-promoting effect of the compound of Example 7 in rat dorsal root ganglion primary culture cells that induced neuronal axonal damage. Specific details for implementing the invention

[0052] The following terms used in this specification are as defined below unless specifically noted otherwise.

[0053] A cyclic amine derivative according to one embodiment of the present invention is characterized by being represented by the following general formula (I).

[0054]

[0055] [In the formula, carbons marked with * are non-carbons, and A represents a group represented by the following general formula (IIa) or (IIb), and

[0056]

[0057] R 1 represents a methyl group, n-propyl group, isopropyl group, 2-methoxyethyl group, 2-ethoxyethyl group, difluoromethyl group, or 3,3,3-trifluoropropyl group, and R 2 represents a hydrogen atom, fluorine atom, or chlorine atom, and R 3 Each independently represents a methyl or ethyl group, and X is -O- or -N(R 3 )- represents]

[0058] In the above cyclic amine derivative, it is preferable that A is a group represented by the general formula (IIa), and then R 1It is preferable that the silver be a methyl group, n-propyl group, isopropyl group, 2-methoxyethyl group, or 3,3,3-trifluoropropyl group.

[0059] In the above-described cyclic amine derivative, it is preferable that A is a group represented by the general formula (IIb), and then X is -N(R 3 It is desirable that )- is, and R 1 It is preferable that the silver be a methyl group, an n-propyl group, an isopropyl group, a 2-methoxyethyl group, or a 3,3,3-trifluoropropyl group.

[0060] In the above-mentioned cyclic amine derivative, R 2 It is preferable that it be a hydrogen atom or a chlorine atom.

[0061] In the above cyclic amine derivative, it is preferable that the stereochemistry of the sub-carbon with * attached is the S configuration.

[0062] In one embodiment according to the above-described cyclic amine derivative of the present invention, A is a group represented by the general formula (IIa), and R 1 is a methyl group, n-propyl group, isopropyl group, 2-methoxyethyl group, or 3,3,3-trifluoropropyl group, and R 2 is a hydrogen atom or a chlorine atom, and R 3 It is a methyl group or an ethyl group. In this embodiment, it is preferable that the stereochemistry of the carbon with * attached is an S configuration.

[0063] In one embodiment according to the above-described cyclic amine derivative of the present invention, A is a group represented by the general formula (IIb), and X is -N(R 3 )-and, R 1 is a methyl group, n-propyl group, isopropyl group, 2-methoxyethyl group, or 3,3,3-trifluoropropyl group, and R 2 is a hydrogen atom or a chlorine atom, and R 3 It is a methyl group or an ethyl group. In this embodiment, it is preferable that the stereochemistry of the carbon with * attached is an S configuration.

[0064] In one embodiment according to the above-described cyclic amine derivative of the present invention, A is a group represented by the general formula (IIb), X is -O-, and R 1 is a methyl group, n-propyl group, isopropyl group, 2-methoxyethyl group, or 3,3,3-trifluoropropyl group, and R 2 is a hydrogen atom or a chlorine atom, and R 3 It is a methyl group or an ethyl group. In this embodiment, it is preferable that the stereochemistry of the carbon with * attached is an S configuration.

[0065] In one embodiment according to the above-mentioned cyclic amine derivative of the present invention, 3-hydroxy-3-(1-methyl-1H-imidazole-2-yl)-1-(4-morpholinopiperidin-1-yl)propan-1-one, 1-(4-(dimethylamino)piperidin-1-yl)-3-(1-propyl-1H-imidazole-2-yl)-3-hydroxypropan-1-one, 1-(4-(dimethylamino)piperidin-1-yl)-3-(1-isopropyl-1H-imidazole-2-yl)-3-hydroxypropan-1-one, 3-(5-chloro-1-methyl-1H-imidazole-2-yl)-1-(4-(dimethylamino)piperidin-1-yl)-3-hydroxypropan-1-one, It is one compound selected from the group consisting of 1-(4-(dimethylamino)piperidin-1-yl)-3-(1-(2-methoxyethyl)-1H-imidazole-2-yl)-3-hydroxypropan-1-one and 1-(4-(dimethylamino)piperidin-1-yl)-3-(1-(3,3,3-trifluoropropyl)-1H-imidazole-2-yl)-3-hydroxypropan-1-one.

[0066] Preferred specific examples of a cyclic amine derivative (hereinafter, cyclic amine derivative (I)) represented by the above general formula (I) are shown in Tables 1 and 2, but the present invention is not limited to these.

[0067]

[0068]

[0069] In addition, if isomers such as enantiomers or stereoisomers exist in the cyclic amine derivative (I), either isomer and a mixture thereof are included in the cyclic amine derivative (I). In addition, if isomers such as enantiomers or stereoisomers exist in the cyclic amine derivative (I), the cyclic amine derivative (I) may be a mixture containing either isomer or a mixture thereof. In addition, if isomers formed by conformation exist in the cyclic amine derivative (I), either isomer and a mixture thereof are included in the cyclic amine derivative (I). The desired isomer can be obtained by a known method or a method equivalent thereto. For example, if enantiomers exist in the cyclic amine derivative (I), one enantiomer separated from the cyclic amine derivative (I) is also included in the cyclic amine derivative (I).

[0070] The desired enantiomer can be obtained by known means (e.g., using an optically active synthetic intermediate, or using a known method or a method equivalent thereto (e.g., optical splitting)) with respect to a racemic mixture of the final product.

[0071] Additionally, the cyclic amine derivative (I) includes a prodrug of the cyclic amine derivative (I) or a pharmacologically acceptable salt thereof. A prodrug of the cyclic amine derivative (I) is a compound that is converted into the cyclic amine derivative (I) enzymatically or chemically in vivo. The active body of the prodrug of the cyclic amine derivative (I) is the cyclic amine derivative (I), but the prodrug of the cyclic amine derivative (I) itself may have activity.

[0072] Examples of prodrugs for cyclic amine derivatives (I) include compounds in which the hydroxyl group of the cyclic amine derivative (I) is alkylated, phosphorylated, or boricated. These compounds can be synthesized from the cyclic amine derivative (I) according to known methods.

[0073] In addition, the prodrug of the cyclic amine derivative (I) may be transformed into the cyclic amine derivative (I) under physiological conditions as described in the known literature ("Development of Medicines," Hirokawa Shoten, 1990, Vol. 7, pp. 163–198 and Progress in Medicine, Vol. 5, 1985, pp. 2157–2161).

[0074] The cyclic amine derivative (I) may be labeled with an isotope, and the labeled isotope may be, for example, 2 H, 3 H, 13 C, 14 C, 15 N, 15 O and / or 18 O is listed.

[0075] Pharmacologically acceptable salts of cyclic amine derivatives (I) are listed as, for example, inorganic salts such as hydrochloride, sulfate, phosphate, or hydrobromide, or organic salts such as oxalate, malonicate, citrate, fumarate, lactate, malate, succinate, tartrate, acetate, trifluoroacetate, maleate, gluconate, benzoate, salicylate, xinafoate, pamoate, ascorbate, adipose, methanesulfonate, p-toluenesulfonate, or cinnamate.

[0076] Cyclic amine derivatives (I) or pharmacologically acceptable salts thereof include their hydrates and solvates.

[0077] Where crystal polymorphs exist in the cyclic amine derivative (I) or its pharmacologically acceptable salt, all crystal polymorphs and mixtures thereof are included in the cyclic amine derivative (I) or its pharmacologically acceptable salt.

[0078] Cyclic amine derivatives (I) or pharmacologically acceptable salts thereof can be synthesized, for example, by following the method described in the known literature (International Publication No. 2016 / 136944).

[0079] "Advilin" includes isoforms, analogs, variants, fragments, or functional derivatives. Advilin is a transcriptional product of the advillin gene, sometimes referred to by the aliases p92 or AVIL, and is a member of the gelsolin family. In humans, two isoforms of advancelin exist, and it is a protein composed of 819 amino acids or 821 amino acids.

[0080] "Advilin" is a molecule that acts on actin filaments and has the function of cleaving double helix-structured actin filaments, as shown in Figure 1.

[0081] "Adbilin function promotion" refers to the maintenance, sustainment, enhancement, and / or normalization of the biological functions of adbilin. Furthermore, adbilin function promotion includes improving and / or normalizing abnormalities in the regulation of actin filament metabolic turnover. Therefore, in situations where actin filaments are increased due to peripheral nerve axonal damage, adbilin function promotion leads to a decrease in actin filaments and an increase in the relative amount of monomeric actin. Additionally, adbilin function promotion includes facilitating nerve axonal extension.

[0082] "Adbilin complex" refers to an aggregate formed by adbilin and molecules, such as proteins, that interact with and / or bind to adbilin.

[0083] The binding of a cyclic amine derivative (I) to Advilin and / or Advilin complexes can be evaluated, for example, by affinity purification using magnetic microparticles, such as FG beads (registered trademark), as described in the known literature (Aono et al., 2018, Biochemical and Biophysical Research Communications, Vol. 505, pp. 1203-1210). In addition to immunoprecipitation using antibodies that recognize Advilin and / or Advilin complexes or affinity purification, the interaction of molecules can be evaluated by analyzing surface plasmon resonance (SPR).

[0084] "Actin filament metabolic turnover" refers to a bidirectional metabolic turnover involving the polymerization of actin monomers and the depolymerization of actin filaments.

[0085] "Abnormal regulation of actin filament metabolic rotation" refers to a condition in which the balance between actin polymerization and actin filament depolymerization in the aforementioned "actin filament metabolic rotation" is disrupted, resulting in a unidirectional process becoming dominant.

[0086] "Improvement of abnormal actin filament metabolic turnover regulation" means normalizing or bringing near-normal the state in which the unidirectional process is dominant in the aforementioned "abnormal actin filament metabolic turnover regulation." In other words, it means normalizing or bringing near-normal the amount of actin filaments and the ratio of the amount of actin filaments to the amount of monomeric actin.

[0087] Whether a cyclic amine derivative (I) or a pharmacologically acceptable salt thereof has an effect of improving abnormalities in the regulation of actin filament metabolic turnover can be evaluated by measuring the amount of actin filaments within cells using cells (e.g., human primary culture dorsal root ganglion neurons, primary culture dorsal root ganglion neurons derived from mammals such as rats and mice, rat adrenochromocytic cell-derived PC12 cells or rat dorsal root ganglion-derived F11 cells, etc.). As a method for measuring the amount of actin filaments within cells, for example, it can be measured by fluorescent staining using fluorescently labeled phalloidins that bind to actin filaments (Carlson et al., NeuroToxicology, 2001, Vol. 22, pp. 819-827).

[0088] In addition, "intracellular actin filament reduction effect" means an effect of reducing the amount of intracellular actin filaments compared to a case where no measures to reduce the amount of intracellular actin filaments are taken, and preferably, an effect of reducing the amount of intracellular actin filaments by 5% or more, 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, or 100% compared to a case where no measures to reduce the amount of intracellular actin filaments are taken.

[0089] In addition, whether a cyclic amine derivative (I) or a pharmacologically acceptable salt thereof has an effect of improving abnormalities in the regulation of actin filament metabolic turnover can be evaluated by measuring the ratio of the amount of actin filaments to the amount of monomeric actin using samples of diseases related to abnormalities in the regulation of actin filament metabolic turnover, such as tissues from patients with neuronal axonal injury or animal models of neuronal axonal injury. As an animal model of neuronal axonal injury, the rat spinal cord ligation model is listed as an example (Kim et al., Pain, 1992, Vol. 50, pp. 355-363). The ratio of the amount of actin filaments to the amount of monomeric actin can be calculated by separating actin filaments and monomeric actin by ultracentrifugation of tissue or cell lysate, and then quantifying each actin by Rotwestern blot (Kim et al., The Journal of Physiology, 2015, Vol. 593, pp. 1873-1886).

[0090] A "nerve axon" refers to a long projection extending from a nerve cell body, also known as a nerve process. Nerve axons form nerve fibers. The terminals of nerve axons branch out and junction with the next nerve cell or target tissue to transmit nerve impulses.

[0091] "Nerve axonal injury" refers to a condition in which a nerve axon is partially or completely destroyed or severed, causing the axon on the tissue side to degenerate or detach from the site of injury.

[0092] "Neural axonal extension" refers to the extension of the axon of a nerve cell. By promoting neural axonal extension, it is possible to expect the resynapse of the axon with the target tissue and / or the reconstruction of neural circuits following neural axonal damage.

[0093] In addition, causes of nerve axon damage are not limited to the following, but include, for example, surgical procedures, toxins, circulatory disorders, trauma resulting from traffic accidents, or radiation therapy.

[0094] Exogenous and endogenous neurotoxins are listed as the toxins mentioned above. For example, exogenous toxins include tetrodotoxin, batrachotoxin, maurotoxin, azitoxin, carbidotoxin, magatoxin, slotoxin, schiratoxin, heftoxin, calciceptin, tygatoxin, or calcyclodin. Endogenous toxins include glutamic acid, N-methyl-D-aspartic acid (NMDA), or caisoacid. In addition, gases (e.g., carbon monoxide), metals (e.g., mercury), methanol, or ethanol are listed. Furthermore, pesticides such as herbicides or insecticides (e.g., rotenone or paraquat) are also listed as toxins. Moreover, in Japan, under the Toxic and Hazardous Substances Control Act, a list of toxins and hazardous substances harmful to the human body is disclosed, and the toxins listed therein are also listed.

[0095] Diseases involving nerve axonal damage (including causative diseases) are not limited to the following, but include, for example, carpal tunnel syndrome, pronator syndrome, anterior interosseous nerve palsy, ulnar nerve canal syndrome (Guillon's canal syndrome), elbow tunnel syndrome, posterior interosseous nerve palsy, radial nerve palsy, thoracic outlet syndrome, axillary nerve palsy, suprascapular nerve palsy, dorsiflexor syndrome, lumbar plexus palsy, plantar spinal canal syndrome, femoral nerve palsy, sciatic nerve palsy, tibial nerve palsy, common peroneal nerve palsy, deep peroneal nerve palsy (forefoot tunnel syndrome), saphenous nerve palsy (Hunter's canal syndrome), cervical spinal stenosis, lumbar spinal stenosis, facial nerve palsy, oculomotor nerve palsy, trochlear nerve palsy, abducens nerve palsy, giant cell vasculitis, Takayasu's arteritis, nodular polyarteritis, Kawasaki disease, polyangiitis Granuloma (Wegener's granulomatosis), microscopic polyvasculitis, eosinophilic polyvasculitic granulomatosis (allergic granulomatous vasculitis, Churk-Strauss syndrome), cloglobulinemia, IgA vasculitis (Henoch-Schonlein purpura), cutaneous leukocytic vasculitis, systemic erythematosus, Sjögren's syndrome, rheumatoid arthritis, mixed connective tissue disease, polymyositis, dermatomyositis, scleroderma, Behcet's disease, Bell's palsy, Ramsay Hunt syndrome, bacterial and viral infections (Lyme disease, HIV infection or Hansen's disease, etc.), sarcoidosis, malignant tumors, etc. are listed.

[0096] Whether a cyclic amine derivative (I) or a pharmacologically acceptable salt thereof has a neuronal axon stretching-promoting effect can be evaluated by measuring the length or density of the stretched neuronal processes using tissue with damaged axons of human neurons, an animal model of neuronal axonal damage, or neurons (e.g., neurons derived from the cerebral cortex or dorsal root ganglion-derived cells, etc.) (Yang et al., Free Radical Biology and Medicine, 2018, Vol. 120, pp. 13-24).

[0097] Cyclic amine derivatives (I) or pharmacologically acceptable salts thereof may be used as medicines for the treatment or prevention of neuronal axonal damage in mammals (e.g., mice, rats, hamsters, rabbits, cats, dogs, cattle, sheep, monkeys, or humans), especially in humans.

[0098] When a cyclic amine derivative (I) or a pharmacologically acceptable salt thereof is used as a medicine, the cyclic amine derivative (I) or a pharmacologically acceptable salt thereof may be administered orally or parenterally, either as is or in combination with a carrier that is acceptable as a medicine.

[0099] For oral administration of a drug containing a cyclic amine derivative (I) or a pharmacologically acceptable salt thereof as an active ingredient, formulations such as tablets (including coated tablets and film-coated tablets), pills, granules, powders, capsules (including soft capsules and microcapsules), syrups, emulsions, or suspensions are listed. In addition, for parenteral administration of a drug containing a cyclic amine derivative (I) or a pharmacologically acceptable salt thereof as an active ingredient, formulations such as injections, infusions, drops, suppositories, topical agents, or patches are listed. In addition, it is also effective to form a sustained-release formulation by combining a cyclic amine derivative (I) or a pharmacologically acceptable salt thereof with a suitable base (e.g., a polymer of butyric acid, a polymer of glycolic acid, a copolymer of butyric acid and glycolic acid, a mixture of a polymer of butyric acid and a polymer of glycolic acid, or a polyglycerol fatty acid ester).

[0100] The formulation of the above-mentioned formulation may be prepared according to known manufacturing methods generally used in the field of formulations. In this case, if necessary, it may be prepared by including excipients, binders, lubricants, disintegrants, sweeteners, surfactants, suspending agents, or emulsifiers generally used in the field of formulations.

[0101] Tablets may be prepared by including, for example, excipients, binders, disintegrants, or lubricants. Pills and granules may be prepared by including, for example, excipients, binders, or disintegrants. In addition, powders and capsules may be prepared by including, for example, excipients. Syrups may be prepared by including, for example, sweeteners. Emulsions or suspensions may be prepared by including, for example, surfactants, suspending agents, or emulsifiers.

[0102] Examples of excipients include lactose, glucose, starch, sucrose, microcrystalline cellulose, licorice powder, mannitol, sodium bicarbonate, calcium phosphate, or calcium sulfate.

[0103] Examples of binders include starch solution, gum arabic solution, gelatin solution, tragant solution, carboxymethylcellulose solution, sodium alginate solution, or glycerin.

[0104] Examples of disintegrants include starch or calcium carbonate.

[0105] Examples of lubricants include magnesium stearate, stearic acid, calcium stearate, or refined talc.

[0106] Examples of sweeteners include glucose, fructose, invert sugar, sorbitol, xylitol, glycerin, or sweet syrup.

[0107] Examples of surfactants include sodium lauryl sulfate, polysorbate 80, sorbitan mono fatty acid ester, or stearic acid polyoxyl 40.

[0108] Examples of suspending agents include gum arabic, sodium alginate, sodium carboxymethylcellulose, methylcellulose, or bentonite.

[0109] Examples of emulsifiers include gum arabic, tragant, gelatin, or polysorbate 80.

[0110] In addition, when preparing a medicine containing a cyclic amine derivative (I) or a pharmacologically acceptable salt thereof as an active ingredient in the above formulation, coloring agents, preservatives, fragrances, stimulants, stabilizers, or viscosity enhancers commonly used in the field of formulation may be added.

[0111] The daily dosage of a drug containing a cyclic amine derivative (I) or a pharmacologically acceptable salt thereof as an active ingredient varies depending on the patient's condition or body weight, the type of compound, or the route of administration, but for example, when administered orally to an adult (body weight about 60 kg), it is preferable to administer the cyclic amine derivative (I) or a pharmacologically acceptable salt thereof in 1 to 3 divided doses in the range of 1 to 1000 mg as an active ingredient, and when administered parenterally to an adult (body weight about 60 kg), if it is an injectable drug, it is preferable to administer the cyclic amine derivative (I) or a pharmacologically acceptable salt thereof by intravenous injection in the range of 0.01 to 100 mg as an active ingredient per 1 kg of body weight.

[0112] Cyclic amine derivatives (I) or pharmacologically acceptable salts thereof may be combined or used in combination with other drugs in appropriate amounts to supplement or enhance therapeutic or preventive effects, or to reduce the dosage. Cyclic amine derivatives (I) or pharmacologically acceptable salts thereof may be administered simultaneously with other drugs or administered sequentially in any order. Other drugs may be used in combination with drugs that treat neuronal axonal damage, although not limited to these.

[0113] Examples

[0114] The present invention will be described in detail below using examples, comparative examples, and reference examples, but the present invention is not limited thereto.

[0115] In the following description, the solvent name indicated in the NMR data represents the solvent used for measurement. Additionally, the 400 MHz NMR spectrum was measured using a JNM-AL Type 400 nuclear magnetic resonance system (manufactured by JEOL Ltd.). Chemical shifts are expressed as δ (unit: ppm) with respect to tetramethylsilane, and signals are represented as s (monuline), d (duuline), t (triuline), q (quaduline), quint (pentuline), sept (hyptuline), m (multiple), br (spread), dd (duuline double), dt (duuline triple), ddd (duuline double), dq (duuline quadruline), td (tripuline double), and tt (tripuline triple). ESI-MS spectra were measured using the Agilent Technologies 1200 Series, G6130A (manufactured by Agilent Technologies). All solvents used were commercially available. Flash column chromatography was performed using the YFLC W-prep2XY (manufactured by Yamahazen).

[0116] The raw materials and intermediates of the cyclic amine derivative (I) were synthesized by the method described in the reference examples below. In addition, for compounds used in the synthesis of the reference example compounds for which the synthesis method was not described, commercially available compounds were used.

[0117] (Reference Example 1) Synthesis of benzyl(S)-2-(3-(4-(dimethylamino)piperidine-1-yl)-1-(1-methyl-1H-imidazole-2-yl)-3-oxopropoxy)acetate:

[0118]

[0119] Sodium hydride (55%, 0.0202 mg, 0.464 mmol) was added to a tetrahydrofuran (0.800 mL) solution of (S)-1-(4-(dimethylamino)piperidin-1-yl)-3-hydroxy-3-(1-methyl-1H-imidazole-2-yl)propan-1-one (0.100 g, 0.357 mmol) at 0°C. After stirring for 10 minutes at the same temperature, benzyl bromoacetate (0.0620 mL, 0.390 mmol) was added to the reaction mixture, and the mixture was stirred for 15 more hours at room temperature. An aqueous ammonium chloride solution was added to the reaction mixture, and the mixture was extracted with chloroform. The organic layer was washed with a 10% aqueous sodium chloride solution, dried with anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was purified by flash column chromatography (NH silica gel, chloroform / methanol), and benzyl(S)-2-(3-(4-(dimethylamino)piperidin-1-yl)-1-(1-methyl-1H-imidazole-2-yl)-3-oxopropoxy)acetate (0.568 g, 0.133 mmol, 37%) was obtained as a colorless oily substance.

[0120] 1 H-NMR (400 MHz, CDCl3) δ: 1.30-1.57(2H, m), 1.73-1.90(2H, m), 2.24-2.37(7H, m), 2.48-2.58(1H, m), 2.95-3.13(2H, m), 3.28-3.36(1H, m), 3.70(3H, d, J=1.6 Hz), 3.93-4.17(3H, m), 4.48-4.54(1H, m), 5.12-5.14(2H, m), 5.30-5.36(1H, m), 6.80(1H, s), 6.96(1H, s), 7.31-7.39(5H, m).

[0121] ESI-MS: m / z=429(M+H) + .

[0122] (Reference Example 2) Synthesis of ethyl 1-methyl-1H-imidazole-2-carboxylate:

[0123]

[0124] Triethylamine (3.40 mL, 24.4 mmol) and ethyl chloroformate (2.34 mL, 24.4 mmol) were added to an acetonitrile (4.0 mL) solution of 1-methyl-1H-imidazole (1.00 g, 12.2 mmol) at 0°C, and the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was filtered using Cerite, and the filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography (silica gel, hexane / ethyl acetate), and ethyl 1-methyl-1H-imidazole-2-carboxylate (1.50 g, 9.73 mmol, 80%) was obtained as a white solid.

[0125] 1 H-NMR (400 MHz, CDCl3) δ: 1.42 (3H, t, J=7.2 Hz), 4.01 (3H, s), 4.40 (2H, q, J=7.2 Hz), 7.01-7.03 (1H, m), 7.13-7.15 (1H, m).

[0126] ESI-MS: m / z=155(M+H) + .

[0127] (Reference Example 3) Synthesis of ethyl 3-(1-methyl-1H-imidazole-2-yl)-3-oxopropanoate:

[0128]

[0129] Sodium hydroxide aqueous solution (1.0N, 14.6mL, 14.6 mmol) was added at room temperature to a methanol (15.0mL) solution of ethyl 1-methyl-1H-imidazole-2-carboxylate (1.50g, 9.73 mmol), and the reaction mixture was stirred at the same temperature for 3 hours. The reaction mixture was cooled to 0°C. Hydrochloric acid (1.0N) was added to the reaction mixture to neutralize it, and then concentrated under reduced pressure. It was azeotropic with toluene, and ethanol was added. The precipitate was filtered using Cerite, and the filtrate was concentrated under reduced pressure. Acetonitrile (7.0mL) and carbonyldiimidazole (1.54g, 9.52 mmol) were added to the obtained crude product at room temperature, and the reaction mixture was stirred at the same temperature for 2.5 hours (Reaction Mixture A). Separately, magnesium chloride (0.997 g, 10.5 mmol) was dissolved in acetonitrile (7.0 mL), ethyl potassium malonicate (1.70 g, 9.99 mmol) and triethylamine (2.98 mL, 21.4 mmol) were added at room temperature, and the reaction mixture was stirred at the same temperature for 2.5 hours (reaction mixture B). Reaction mixture A was added to reaction mixture B at room temperature, and the reaction mixture was stirred at 80°C for 2 hours. The reaction mixture was cooled to room temperature. After adding hydrochloric acid (1.0 N) to the reaction mixture, it was extracted with ethyl acetate. The organic layer was washed with a 10% aqueous sodium chloride solution, dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography (silica gel, hexane / ethyl acetate), and ethyl 3-(1-methyl-1H-imidazole-2-yl)-3-oxopropanoate (0.721 g, 3.67 mmol, 38%) was obtained as a white solid.

[0130] 1 H-NMR (400 MHz, CDCl3) δ: 1.27(3H, t, J=7.2 Hz), 4.01(3H, s), 4.13(2H, s), 4.21(2H, q, J=7.2 Hz), 7.05-7.07(1H, m), 7.15-7.17(1H, m).

[0131] ESI-MS: m / z=197(M+H) + .

[0132] (Reference Example 4) Synthesis of 4-(morpholin-4-yl)piperidine:

[0133]

[0134] Morpholine (0.792 g, 9.09 mmol), sodium triacetoxyborohydride (1.93 g, 9.09 mmol), and acetic acid (0.0460 g, 0.758 mmol) were added to a dichloromethane (25.0 mL) solution of 1-tert-butoxycarbonyl-4-piperidinone (1.51 g, 7.58 mmol) at 0°C, and the mixture was stirred at room temperature for 16 hours. The reaction mixture was cooled to 0°C. A saturated sodium bicarbonate aqueous solution was added to the reaction mixture, and it was extracted with dichloromethane. The organic layer was dried with anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was dissolved in hydrochloric acid (1.0 N) and extracted with ethyl acetate. A 48% sodium hydroxide aqueous solution was added to the aqueous layer to make it basic, and then extracted with dichloromethane. The organic layer was dried with anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was dissolved in methanol (25.0 mL), concentrated hydrochloric acid (5.0 mL) was added, and the mixture was stirred at 40°C for 12 hours. The reaction mixture was concentrated and dried, then dissolved in distilled water. A 48% aqueous sodium hydroxide solution was added to make it basic, and then extracted with dichloromethane. The organic layer was dried with anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. 4-(morpholine-4-yl)piperidine (1.52 g, 5.63 mmol, 74%) was obtained as a yellow solid.

[0135] 1 H-NMR (400 MHz, CDCl3) δ: 1.34 (2H, dd, J=12.0, 4.0 Hz), 1.40 (2H, dd, J=12.0, 4.0 Hz), 1.85 (2H, d, J=12.4 Hz), 2.28 (1H, tt, J=11.2, 4.0 Hz), 3.53-3.63(6H, m), 3.15(2H, d, J=12.4 Hz), 3.73(4H, t, J=4.4 Hz).

[0136] ESI-MS: m / z=171(M+H) +

[0137] (Reference Example 5) Synthesis of 1-(1-methyl-1H-imidazole-2-yl)-3-(4-morpholinopiperidine-1-yl)propane-1,3-dione:

[0138]

[0139] 4-(morpholine-4-yl)piperidine (0.158 g, 0.928 mmol) was added at room temperature to a toluene (0.460 mL) solution of ethyl 3-(1-methyl-1H-imidazole-2-yl)-3-oxopropanoate (0.200 g, 1.02 mmol), and the reaction mixture was stirred at 110°C for 16 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by flash chromatography (silica gel, chloroform / methanol), and 1-(1-methyl-1H-imidazole-2-yl)-3-(4-morpholineopiperidine-1-yl)propane-1,3-dione (0.285 g, 0.890 mmol, 96%) was obtained as a colorless oily product.

[0140] 1 H-NMR (400 MHz, CDCl3) δ: 1.40-1.64(2H, m), 1.82-1.92(2H, m), 2.37-2.47(1H, m), 2.52-2.58(4H, m), 3.05-3.15(1H, m), 3.69-3.76(5H, m), 3.82-3.90(1H, m), 4.01(3H, s), 4.16-4.31(2H, m), 4.58-4.65(1H, m), 7.04-7.06(1H, m), 7.13-7.15(1H, m).

[0141] ESI-MS: m / z=321(M+H) + .

[0142] (Reference Example 6) Synthesis of 1-propyl-1H-imidazole-2-carbaldehydride:

[0143]

[0144] 1-iodopropane (1.22 mL, 12.5 mmol) and potassium carbonate (2.16 g, 15.6 mmol) were added to a 10.0 mL N, N-dimethylformamide solution of 1H-imidazole-2-carbaldehydride (1.00 g, 10.4 mmol) and stirred at 60°C for 3 hours. Water was added to the reaction mixture and extracted with ethyl acetate. The organic layer was washed with a 10% aqueous sodium chloride solution, dried with anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel, hexane / ethyl acetate), and 1-propyl-1H-imidazole-2-carbaldehydride (0.786 g, 5.69 mmol, 55%) was obtained as a yellow oily substance.

[0145] 1 H-NMR (400 MHz, CDCl3) δ: 0.93 (3H, t, J=7.4 Hz), 1.77-1.85 (2H, m), 4.37 (2H, t, J=7.2 Hz), 7.16 (1H, s), 7.28 (1H, s), 9.82 (1H, s).

[0146] (Reference Example 7) Synthesis of 1-Isopropyl-1H-Imidazole-2-Carvaldehyde:

[0147]

[0148] 2-iodopropane (1.26 mL, 12.5 mmol) and potassium carbonate (2.16 g, 15.6 mmol) were added to a 10 mL N, N-dimethylformamide solution of 1H-imidazole-2-carbaldehydride (1.00 g, 10.4 mmol) and stirred at 60°C for 3 hours. Water was added to the reaction mixture and extracted with ethyl acetate. The organic layer was washed with a 10% aqueous sodium chloride solution, dried with anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel, hexane / ethyl acetate), and 1-isopropyl-1H-imidazole-2-carbaldehydride (0.703 g, 5.09 mmol, 49%) was obtained as a yellow oily substance.

[0149] 1H-NMR (400 MHz, CDCl3) δ: 1.47 (6H, t, J=6.6 Hz), 5.48 (1H, q, J=6.6 Hz), 7.30 (1H, s), 7.33 (1H, s), 9.83 (1H, s).

[0150] (Reference Example 8) Synthesis of 5-chloro-1-methyl-1H-imidazole-2-carbaldehydride:

[0151]

[0152] Desmartin reagent (1.04 g, 2.46 mmol) was added to a solution of (5-chloro-1-methyl-1H-imidazole-2-yl)methanol (0.300 g, 2.05 mmol) in dichloromethane (20.0 mL) at 0°C and stirred at the same temperature for 3 hours. A 10% aqueous sodium thiosulfate solution and a saturated aqueous sodium bicarbonate solution were added to the reaction mixture, and the mixture was extracted with chloroform. The organic layer was washed with a 10% aqueous sodium chloride solution, dried with anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel, hexane / ethyl acetate), and 5-chloro-1-methyl-1H-imidazole-2-carbaldehydride (0.235 g, 1.62 mmol, 79%) was obtained as a white solid.

[0153] 1 H-NMR (400 MHz, CDCl3) δ: 3.98(3H, s), 7.24(1H, s), 9.70(1H, s).

[0154] (Reference Example 9) Synthesis of 1-(2-methoxyethyl)-1H-imidazole-2-carbaldehydride:

[0155]

[0156] 2-bromoethylmethyl ether (1.20 mL, 12.5 mmol), potassium carbonate (2.16 g, 15.6 mmol), and sodium iodide (0.468 g, 3.12 mmol) were added to a 1H-imidazole-2-carbaldehydride (1.00 g, 10.4 mmol) solution of N,N-dimethylformamide (10.0 mL), and stirred at 60°C for 3 hours. Water was added to the reaction mixture, and it was extracted with ethyl acetate. The organic layer was washed with a 10% aqueous sodium chloride solution, dried with anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel, hexane / ethyl acetate), and 1-(2-methoxyethyl)-1H-imidazole-2-carbaldehydride (0.535 g, 3.47 mmol, 33%) was obtained as a white solid.

[0157] 1 H-NMR (400 MHz, CDCl3) δ: 3.32 (3H, s), 3.67 (2H, t, J=5.0 Hz), 4.59 (2H, t, J=5.0 Hz), 7.23-7.30 (2H, m), 9.81 (1H, s).

[0158] (Reference Example 10) Synthesis of 1-(3,3,3-trifluoropropyl)-1H-imidazole-2-carbaldehydride:

[0159]

[0160] 1,1,1-trifluoro-3-iodopropane (0.710 mL, 6.24 mmol) and potassium carbonate (1.08 g, 7.81 mmol) were added to a solution of 1H-imidazole-2-carbaldehydride (0.500 g, 5.20 mmol) in N,N-dimethylformamide (5.20 mL) and stirred at 60°C for 5 hours. Water was added to the reaction mixture and extracted with ethyl acetate. The organic layer was washed with a 10% aqueous sodium chloride solution, dried with anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel, hexane / ethyl acetate), and 1-(3,3,3-trifluoropropyl)-1H-imidazole-2-carbaldehydride (0.0863 g, 0.449 mmol, 8.6%) was obtained as a colorless oily substance.

[0161] 1 H-NMR (400 MHz, CDCl3) δ: 2.60-2.72(2H, m), 4.61(2H, t, J=6.8 Hz), 7.18(1H, s), 7.32(1H, s), 9.83(1H, s).

[0162] (Reference Example 11) Synthesis of 1-(4-(dimethylamino)piperidine-1-yl)ethanolone:

[0163]

[0164] Pyridine (0.922 mL, 9.75 mmol) and acetic anhydride (0.946 mL, 11.7 mmol) were added to a dichloromethane (7.8 mL) solution of 4-dimethylaminopiperidine (1.00 g, 7.79 mmol) at 0°C, and the reaction mixture was stirred at room temperature for 16 hours. An aqueous solution of saturated sodium bicarbonate was added to the reaction mixture, and it was extracted with chloroform. The organic layer was washed with a 10% aqueous sodium chloride solution, dried with anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was purified by flash column chromatography (NH4 silica gel, chloroform / methanol), and 1-(4-(dimethylamino)piperidine-1-yl)ethanolone (0.869 g, 6.78 mmol, 87%) was obtained as a colorless oily substance.

[0165] 1 H-NMR (400 MHz, CDCl3) δ: 1.30-1.47(2H, m), 1.79-1.92(2H, m), 2.10(3H, s), 2.25-2.40(7H, m), 2.53-2.63(1H, m), 3.01-3.11(1H, m), 3.81-3.90(1H, m), 4.58-4.66(1H, m) .

[0166] ESI-MS: m / z=171(M+H) + .

[0167] (Reference Example 12) Synthesis of 4-(1-methylpiperazine-4-yl)piperidine:

[0168]

[0169] To a dichloromethane (25.0 mL) solution of 1-tert-butoxycarbonyl-4-piperidinone (1.50 g, 7.53 mmol) in which 1-methylpiperazine (0.905 g, 9.03 mmol), acetic acid (0.497 g, 8.28 mmol), and sodium triacetoxyborohydride (1.92 g, 9.03 mmol) were added at 0°C, and the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was cooled to 0°C. A saturated aqueous sodium bicarbonate solution was added to the reaction mixture and extracted with dichloromethane. The organic layer was dried with anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was dissolved in hydrochloric acid (1.0 N) and extracted with ethyl acetate. A 48% aqueous sodium hydroxide solution was added to the aqueous layer to make it basic, and then extracted with dichloromethane. The organic layer was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was dissolved in methanol (25.0 mL), concentrated hydrochloric acid (5.0 mL) was added, and the mixture was stirred at 40°C for 12 hours. The reaction solution was concentrated under reduced pressure and then dissolved in distilled water. After adding a 48% aqueous sodium hydroxide solution to make it basic, it was extracted with dichloromethane. The organic layer was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure to obtain 4-(1-methylpiperazine-4-yl)piperidine (0.826 g, 4.51 mmol, 60%) as a white solid.

[0170] 1 H-NMR (400 MHz, CDCl3) δ: 1.35 (2H, dd, J=12.0, 3.6 Hz), 1.41 (2H, dd, J=12.0, 3.6 Hz), 1.85 (2H, d, J=12.8 Hz), 1.96-2.06 (2H, br), 2.28(3H, s), 2.32(1H, tt, J=11.6, 3.6 Hz), 3.37-3.70(8H, m), 3.14(2H, d, J=12.8 Hz).

[0171] ESI-MS: m / z=169(M+H) + .

[0172] (Reference Example 13) Synthesis of crude 4-diethylaminopiperidine:

[0173]

[0174] Diethylamine (0.276 mL, 2.68 mmol), acetic acid (0.0120 mL, 0.214 mmol), and sodium triacetoxyborohydride (0.681 g, 3.22 mmol) were added to a solution of benzyl 4-oxopiperidine-1-carboxylate (0.500 g, 2.14 mmol) in dichloromethane (12.0 mL) at 0°C, and the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was cooled to 0°C. An aqueous solution of saturated sodium bicarbonate was added to the reaction mixture, and it was extracted with chloroform. The organic layer was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (NH4 silica gel, chloroform / methanol). The obtained crude purified product was dissolved in methanol (8.0 mL), palladium / carbon (10% wet, 0.180 g, 0.169 mmol) was added at room temperature, and the mixture was stirred for 16 hours under a hydrogen atmosphere. The reaction mixture was filtered using cerite, the filtrate was concentrated under reduced pressure, and the crude product of 4-diethylaminopiperidine was obtained.

[0175] (Reference Example 14) Synthesis of 1-(4-(dimethylamino)piperidine-1-yl)-3-(1-methyl-1H-imidazole-2-yl)propane-1,3-dione:

[0176]

[0177] A tetrahydrofuran solution of lithium diisopropylamide (2.0 M, 7.05 mL, 14.1 mmol) was added dropwise to a tetrahydrofuran solution (20 mL) of 1-(4-(dimethylamino)piperidin-1-yl)ethanol (1.00 g, 5.87 mmol) at -78°C and stirred for 1 hour at the same temperature. A tetrahydrofuran solution (9.0 mL) of ethyl 1-methyl-1H-imidazole-2-carboxylate (1.09 g, 7.05 mmol) was added to the reaction mixture at the same temperature and stirred for 1 hour, then stirred for another 1 hour at 0°C. Saturated aqueous ammonium chloride solution and an aqueous potassium carbonate solution were added sequentially to the reaction mixture and extracted with chloroform. After washing the organic layer with a 10% aqueous sodium chloride solution, it was dried with anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was purified by flash column chromatography (NH4 silica gel, hexane / ethyl acetate), and 1-(4-(dimethylamino)piperidine-1-yl)-3-(1-methyl-1H-imidazole-2-yl)propane-1,3-dione (0.990 g, 3.56 mmol, 61%) was obtained as a colorless oily substance.

[0178] 1 H-NMR (400 MHz, CDCl3) δ: 1.32-1.5(2H, m), 1.80-1.94(2H, m), 2.22-41(7H, m), 2.60-2.70(1H, m), 3.03-3.13(1H, m), 3.80-3.89(1H, m), 4.01(3H, s), 4.23(2H, dd, J=15.6, 36.8 Hz), 4.55-4.67(1H, m), 7.05(1H, s), 7.14(1H, s).

[0179] ESI-MS: m / z=279(M+H) + .

[0180] (Reference Example 15) Synthesis of 1-(1-methyl-1H-imidazole-2-yl)-3-(4-(4-methylpiperazine-1-yl)piperidine-1-yl)propane-1,3-dione:

[0181]

[0182] 4-(1-methylpiperazine-4-yl)piperidine (0.170 g, 0.927 mmol) was added to a toluene (0.46 mL) solution of ethyl 3-(1-methyl-1H-imidazole-2-yl)-3-oxopropanoate (0.200 g, 1.02 mmol) at room temperature, and the reaction mixture was stirred at 110°C for 16 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography (NH4 silica gel, chloroform / methanol), and 1-(1-methyl-1H-imidazole-2-yl)-3-(4-(4-methylpiperazine-1-yl)piperidine-1-yl)propane-1,3-dione (0.290 g, 0.870 mmol, 94%) was obtained as a colorless oily product.

[0183] 1 H-NMR (400 MHz, CDCl3) δ: 1.38-1.60(2H, m), 1.82-1.90(2H, m), 1.95-2.10(1H, m), 2.27(3H, s), 2.36-2.68(9H, m), 3.02-3.12(1H, m), 3.79-3.88(1H, m), 3.98(3H, s), 4.13-4.28(2H, m), 4.57-4.90(1H, m), 7.02-7.04(1H, m), 7.11-7.13(1H, m).

[0184] ESI-MS: m / z=334(M+H) + .

[0185] (Reference Example 16) Synthesis of 1-(4-(diethylamino)piperidine-1-yl)-3-(1-methyl-1H-imidazole-2-yl)propane-1,3-dione:

[0186]

[0187] To a toluene (0.38 mL) solution of ethyl 3-(1-methyl-1H-imidazole-2-yl)-3-oxopropanoate (0.150 g, 0.765 mmol), crude 4-diethylaminopiperidine (0.143 g, 0.917 mmol) was added at room temperature, and the reaction mixture was stirred at 110°C for 10 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography (NH4 silica gel, chloroform / methanol), and 1-(4-(diethylamino)piperidine-1-yl)-3-(1-methyl-1H-imidazole-2-yl)propane-1,3-dione (0.0750 g, 0.245 mmol, 32%) was obtained as a colorless oily product.

[0188] 1 H-NMR (400 MHz, CDCl3) δ: 1.02 (6H, t, J=6.8 Hz), 1.37-1.58 (2H, m), 1.73-1.98 (2H, m), 2.48-2.78 (6H, m), 3.01-3.11 (1H, m), 3.80-3.88(1H, m), 4.00(3H, s), 4.14-4.28(2H, m), 4.60-4.70(1H, m), 7.03-7.05(1H, m), 7.12-7.14(1H, m).

[0189] ESI-MS: m / z=307(M+H) + .

[0190] (Reference Example 17) Synthesis of 1-(4-(dimethylamino)piperidine-1-yl)-3-hydroxy-3-(1-methyl-1H-imidazole-2-yl)propan-1-one:

[0191]

[0192] A tetrahydrofuran solution of lithium diisopropylamide (2.0 M, 0.162 mL, 0.323 mmol) was added dropwise to a tetrahydrofuran solution (0.8 mL) of 1-(4-(dimethylamino)piperidin-1-yl)ethanol (0.0500 g, 0.294 mmol) at -78°C and stirred for 1 hour at the same temperature. A tetrahydrofuran solution (0.4 mL) of 1-methyl-1H-imidazole-2-carbaldehydride (0.0390 g, 0.352 mmol) was added to the reaction mixture at the same temperature and stirred for 1 hour, then stirred for another 1 hour at 0°C. Saturated aqueous ammonium chloride solution and aqueous potassium carbonate solution were added sequentially to the reaction mixture and extracted with chloroform. After washing the organic layer with a 10% aqueous sodium chloride solution, it was dried with anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel, chloroform / methanol), and 1-(4-(dimethylamino)piperidin-1-yl)-3-hydroxy-3-(1-methyl-1H-imidazole-2-yl)propan-1-one (0.0220 g, 0.0785 mmol, 27%) was obtained as a colorless oily substance.

[0193] 1 H-NMR (400 MHz, CDCl3) δ: 1.32-1.53(2H, m), 1.82-1.92(2H, m), 2.27-2.41(7H, m), 2.60-2.72(1H, m), 2.98-3.23(3H, m), 3.77(3H, s), 3.99-4.08(1H, m), 4.58-4.82(2H, m), 5.18-5.26(1H, m), 6.86(1H, s), 6.93(1H, s).

[0194] ESI-MS: m / z=281(M+H) + .

[0195] (Example 1) Synthesis of 3-hydroxy-3-(1-methyl-1H-imidazole-2-yl)-1-(4-morpholinopiperidin-1-yl)propan-1-one:

[0196]

[0197] Sodium borohydride (0.0370 g, 0.979 mmol) was added at room temperature to a methanol (4.50 mL) solution of 1-(1-methyl-1H-imidazole-2-yl)-3-(4-morpholinopiperidin-1-yl)propane-1,3-dione (0.285 g, 0.890 mmol) and the reaction mixture was stirred at the same temperature for 3 hours. A saturated aqueous sodium bicarbonate solution was added to the reaction mixture and concentrated under reduced pressure. Distilled water was added to the residue and extracted with chloroform. The organic layer was washed with a 10% aqueous sodium chloride solution, dried with anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was purified by flash chromatography (silica gel, chloroform / methanol) and 3-hydroxy-3-(1-methyl-1H-imidazole-2-yl)-1-(4-morpholinopiperidin-1-yl)propan-1-one (0.0711 g, 0.221 mmol, 25%) (hereinafter, the compound of Example 1) was obtained as a colorless oily substance.

[0198] 1 H-NMR (400 MHz, DMSO-d6) δ: 1.08-1.50(7H, m), 1.60-1.76(2H, m), 2.36-2.46(5H, m), 2.75-3.05(3H, m), 3.64(3H, s), 3.92-4.02(1H, m), 4.32-4.42(1H, m), 4.99-5.08(1H, m), 5.34-5.49(1H, m), 6.70-6.72(1H, m), 7.01-7.03(1H, m).

[0199] ESI-MS: m / z=323(M+H) + .

[0200] (Example 2) Synthesis of 1-(4-(dimethylamino)piperidine-1-yl)-3-(1-propyl-1H-imidazole-2-yl)-3-hydroxypropan-1-one:

[0201]

[0202] A tetrahydrofuran solution of lithium diisopropylamide (2.0 M, 0.969 mL, 1.94 mmol) was added dropwise to a tetrahydrofuran solution (6.00 mL) of 1-(4-dimethylaminopiperidin-1-yl)ethanol (0.300 g, 1.76 mmol) at -78°C and stirred for 1 hour at the same temperature. A tetrahydrofuran solution (2.8 mL) of 1-propyl-1H-imidazole-2-carbaldehydride (0.292 g, 2.12 mmol) was added to the reaction mixture at the same temperature and stirred for 1 hour, followed by stirring for another 1 hour at 0°C. Saturated aqueous ammonium chloride solution and an aqueous potassium carbonate solution were sequentially added to the reaction mixture and extracted with chloroform. After washing the organic layer with a 10% aqueous sodium chloride solution, it was dried with anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was purified by flash column chromatography (NH4 silica gel, chloroform / methanol), and 1-(4-(dimethylamino)piperidin-1-yl)-3-(1-propyl-1H-imidazole-2-yl)-3-hydroxypropan-1-one (0.296 g, 0.960 mmol, 55%) (hereinafter, the compound of Example 2) was obtained as a colorless oily substance.

[0203] 1 H-NMR (400 MHz, DMSO-d6) δ: 0.85 (3H, t, J=7.4Hz), 1.00-1.40 (2H, m), 1.61-1.80 (4H, m), 2.10-2.33 (7H, m), 2.45-2.59 (1H, m), 2.73-2.88(1H, m), 2.93-3.13(2H, m), 3.86-4.00(3H, m), 4.25-4.35(1H, m), 4.98-5.05(1H, m), 5.34-5.40(1H, m), 6.72(1H, s), 7.07(1H, s).

[0204] ESI-MS: m / z=309(M+H) + .

[0205] (Example 3) Synthesis of 1-(4-(dimethylamino)piperidine-1-yl)-3-(1-isopropyl-1H-imidazole-2-yl)-3-hydroxypropan-1-one:

[0206]

[0207] A tetrahydrofuran solution of lithium diisopropylamide (2.0 M, 0.969 mL, 1.94 mmol) was added dropwise to a tetrahydrofuran solution (6.00 mL) of 1-(4-dimethylaminopiperidin-1-yl)ethanol (0.300 g, 1.76 mmol) at -78°C and stirred for 1 hour at the same temperature. A tetrahydrofuran solution (2.8 mL) of 1-isopropyl-1H-imidazole-2-carbaldehydride (0.292 g, 2.12 mmol) was added to the reaction mixture at the same temperature and stirred for 1 hour, then stirred for another 1 hour at 0°C. A saturated aqueous ammonium chloride solution and an aqueous potassium carbonate solution were added sequentially to the reaction mixture and extracted with chloroform. After washing the organic layer with a 10% aqueous sodium chloride solution, it was dried with anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was purified by flash column chromatography (NH4 silica gel, chloroform / methanol), and 1-(4-(dimethylamino)piperidin-1-yl)-3-(1-isopropyl-1H-imidazole-2-yl)-3-hydroxypropan-1-one (0.302 g, 0.979 mmol, 56%) (hereinafter referred to as the compound of Example 3) was obtained as a colorless oily substance.

[0208] 1 H-NMR (400 MHz, DMSO-d6) δ: 1.04-1.41(8H, m), 1.62-1.80(2H, m), 2.16(6H, s), 2.25-2.34(1H, m), 2.48-2.59(2H, m), 2.76-2.88(1H, m), 2.95-3.16(2H, m), 3.90-4.00(1H, m), 4.27-4.38(1H, m), 5.05-5.12(1H, m), 5.36-5.42(1H, m), 6.77(1H, s), 7.20(1H, s).

[0209] ESI-MS: m / z=309(M+H)+.

[0210] (Example 4) Synthesis of 3-(5-chloro-1-methyl-1H-imidazole-2-yl)-1-(4-(dimethylamino)piperidin-1-yl)-3-hydroxypropan-1-one:

[0211]

[0212] A tetrahydrofuran solution of lithium diisopropylamide (2.0 M, 0.745 mL, 1.49 mmol) was added dropwise to a tetrahydrofuran solution (5.10 mL) of 1-(4-dimethylaminopiperidin-1-yl)ethanol (0.231 g, 1.36 mmol) at -78°C and stirred for 1 hour at the same temperature. A tetrahydrofuran solution (1.70 mL) of 5-chloro-1-methyl-1H-imidazole-2-carbaldehydride (0.235 g, 1.63 mmol) was added to the reaction mixture at the same temperature and stirred for 1 hour, then stirred for another 1 hour at 0°C. Saturated aqueous ammonium chloride solution and aqueous potassium carbonate solution were added sequentially to the reaction mixture and extracted with chloroform. After washing the organic layer with a 10% aqueous sodium chloride solution, it was dried with anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was purified by flash column chromatography (NH4 silica gel, chloroform / methanol), and 3-(5-chloro-1-methyl-1H-imidazole-2-yl)-1-(4-(dimethylamino)piperidin-1-yl)-3-hydroxypropan-1-one (0.159 g, 0.505 mmol, 37%) (hereinafter referred to as the compound of Example 4) was obtained as a colorless oily substance.

[0213] 1 H-NMR (400 MHz, DMSO-d6) δ: 1.04-1.21(1H, m), 1.28-1.40(1H, m), 1.64-1.80(2H, m), 2.15(6H, s), 2.24-2.35(1H, m), 2.44-2.60(1H, m), 2.78-2.88(1H, m), 2.95-3.11(2H, m), 3.59(3H, s), 3.90-3.98(1H, m), 4.27-4.35(1H, m), 5.00-5.10(1H, m), 5.50-5.58(1H, m), 6.85(1H, s).

[0214] ESI-MS: m / z=315(M+H)+.

[0215] (Example 5) Synthesis of 1-(4-(dimethylamino)piperidine-1-yl)-3-(1-(2-methoxyethyl)-1H-imidazole-2-yl)-3-hydroxypropan-1-one:

[0216]

[0217] A tetrahydrofuran solution of lithium diisopropylamide (2.0 M, 0.969 mL, 1.94 mmol) was added dropwise to a tetrahydrofuran solution (6.00 mL) of 1-(4-dimethylaminopiperidin-1-yl)ethanol (0.300 g, 1.76 mmol) at -78°C and stirred for 1 hour at the same temperature. A tetrahydrofuran solution (2.80 mL) of 1-(2-methoxyethyl)-1H-imidazole-2-carbaldehydride (0.292 g, 2.12 mmol) was added to the reaction mixture at the same temperature and stirred for 1 hour, followed by stirring for another 1 hour at 0°C. Saturated aqueous ammonium chloride solution and potassium carbonate solution were added sequentially to the reaction mixture and extracted with chloroform. After washing the organic layer with a 10% aqueous sodium chloride solution, it was dried with anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was purified by flash column chromatography (NH4 silica gel, chloroform / methanol), and 1-(4-(dimethylamino)piperidin-1-yl)-3-(1-(2-methoxyethyl)-1H-imidazole-2-yl)-3-hydroxypropan-1-one (0.193 g, 0.594 mmol, 34%) (hereinafter, the compound of Example 5) was obtained as a colorless oily substance.

[0218] 1H-NMR (400 MHz, DMSO-d6) δ: 1.04-1.40(2H, m), 1.62-1.80(2H, m), 2.10-2.35(7H, m), 2.46-2.59(1H, m), 2.80-2.90(1H, m), 2.95-3.10(2H, m), 3.24(3H, s), 3.61(2H, t, J=5.5 Hz), 3.90-4.00(1H, m), 4.10-4.38(3H, m), 5.05-5.11(1H, m), 5.38-5.42(1H, m), 6.73(1H, s), 7.07(1H, s).

[0219] ESI-MS: m / z=325(M+H)+.

[0220] (Example 6) Synthesis of 1-(4-(dimethylamino)piperidine-1-yl)-3-(1-(3,3,3-trifluoropropyl)-1H-imidazole-2-yl)-3-hydroxypropan-1-one:

[0221]

[0222] A tetrahydrofuran solution of lithium diisopropylamide (2.0 M, 0.246 mL, 0.492 mmol) was added dropwise to a tetrahydrofuran solution (1.80 mL) of 1-(4-dimethylaminopiperidin-1-yl)ethane (0.0760 g, 0.448 mmol) at -78°C and stirred for 1 hour at the same temperature. A tetrahydrofuran solution (0.70 mL) of 1-(3,3,3-trifluoropropyl)-1H-imidazole-2-carbaldehydride (0.0860 g, 0.448 mmol) was added to the reaction mixture at the same temperature and stirred for 1 hour, followed by stirring for another 1 hour at 0°C. Saturated aqueous ammonium chloride solution and potassium carbonate solution were sequentially added to the reaction mixture and extracted with chloroform. After washing the organic layer with a 10% aqueous sodium chloride solution, it was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (NH4 silica gel, chloroform / methanol), and 1-(4-(dimethylamino)piperidine-1-yl)-3-(1-(3,3,3-trifluoropropyl)-1H-imidazole-2-yl)-3-hydroxypropan-1-one (0.0845 g, 0.233 mmol, 52%) (hereinafter, the compound of Example 6) was obtained as a colorless oily substance.

[0223] 1 H-NMR (400 MHz, DMSO-d6) δ: 1.03-1.40 (2H, m), 1.63-1.79 (2H, m), 2.10-2.33 (7H, m), 2.47-2.59 (1H, m), 2.78-2.90 (3H, m), 2.95-3.13(2H, m), 3.90-3.98(1H, m), 4.21-4.36(3H, m), 5.03-5.10(1H, m), 5.49-5.54(1H, m), 6.77(1H, s), 7.17(1H, s).

[0224] ESI-MS: m / z=363(M+H)+.

[0225] (Example 7) Synthesis of (S)-1-(4-(dimethylamino)piperidine-1-yl)-3-hydroxy-3-(1-methyl-1H-imidazole-2-yl)propan-1-one:

[0226]

[0227] 1-(4-(dimethylamino)piperidin-1-yl)-3-hydroxy-3-(1-methyl-1H-imidazole-2-yl)propan-1-one (3.32 g) was optically separated by HPLC purification, and the eluent was concentrated under reduced pressure. The residue was purified by flash column chromatography (NH silica gel, chloroform / methanol), and (S)-1-(4-(dimethylamino)piperidin-1-yl)-3-hydroxy-3-(1-methyl-1H-imidazole-2-yl)propan-1-one (0.467 g, >99%ee) (hereinafter, compound of Example 7) was obtained as a white solid.

[0228] HPLC retention time: 8.4 min, Instrument: LC-10ADvp system manufactured by Shimadzu Corporation, Column: CHIRALCEL OZ-H, 4.6×250 mm (manufactured by Daicel Corporation), Solvent: Methanol containing 0.01% ethylenediamine (v / v), Flow rate: 0.5 mL / min, Detection method: UV 220 nm, Column temperature: 40℃.

[0229] 1 H-NMR (400 MHz, CDCl3) δ: 1.32-1.53(2H, m), 1.82-1.92(2H, m), 2.27-2.41(7H, m), 2.60-2.72(1H, m), 2.98-3.23(3H, m), 3.77(3H, s), 3.99-4.08(1H, m), 4.58-4.82(2H, m), 5.18-5.26(1H, m), 6.86(1H, s), 6.93(1H, s).

[0230] ESI-MS: m / z=281(M+H) + .

[0231] (Example 8) Synthesis of 3-hydroxy-3-(1-methyl-1H-imidazole-2-yl)-1-(4-(4-methylpiperazine-1-yl)piperidin-1-yl)propan-1-one:

[0232]

[0233] Sodium borohydride (0.0360 g, 0.957 mmol) was added to a methanol (4.4 mL) solution of 1-(1-methyl-1H-imidazole-2-yl)-3-(4-(4-methylpiperazine-1-yl)piperidin-1-yl)propane-1,3-dione (0.290 g, 0.870 mmol) at room temperature, and the reaction mixture was stirred at the same temperature for 3 hours. A saturated aqueous sodium bicarbonate solution was added to the reaction mixture and concentrated under reduced pressure. Distilled water was added to the residue and extracted with chloroform. The organic layer was washed with a 10% aqueous sodium chloride solution, dried with anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was purified by flash column chromatography (NH4 silica gel, chloroform / methanol) and 3-hydroxy-3-(1-methyl-1H-imidazole-2-yl)-1-(4-(4-methylpiperazine-1-yl)piperidin-1-yl)propan-1-one (0.140 g, 0.417 mmol, 48%) (hereinafter, compound of Example 8) was obtained as a colorless oily substance.

[0234] 1 H-NMR (400 MHz, DMSO-d6) δ: 1.45-1.66 (4H, m), 1.87-1.95 (2H, m), 2.26-2.30 (3H, s), 2.38-2.70 (8H, m), 2.98-3.23 (3H, m), 3.77 (3H, s), 4.00-4.10(1H, m), 4.60-4.70(2H, m), 5.17-5.25(1H, m), 6.85-6.88(1H, m), 6.92-6.95(1H, m).

[0235] ESI-MS: m / z=336(M+H) + .

[0236] (Example 9) Synthesis of 1-(4-(diethylamino)piperidin-1-yl)-3-hydroxy-3-(1-methyl-1H-imidazole-2-yl)propan-1-one:

[0237]

[0238] Sodium borohydride (0.0109 g, 0.287 mmol) was added to a methanol (1.3 mL) solution of 1-(4-(diethylamino)piperidin-1-yl)-3-(1-methyl-1H-imidazole-2-yl)propane-1,3-dione (0.0800 g, 0.261 mmol) at room temperature, and the reaction mixture was stirred at the same temperature for 3 hours. A saturated sodium bicarbonate aqueous solution was added to the reaction mixture and concentrated under reduced pressure. Distilled water was added to the residue and extracted with chloroform. The organic layer was washed with a 10% sodium chloride aqueous solution, dried with anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was purified by flash column chromatography (NH4 silica gel, chloroform / methanol) and 1-(4-(diethylamino)piperidin-1-yl)-3-hydroxy-3-(1-methyl-1H-imidazole-2-yl)propan-1-one (0.0561 g, 0.182 mmol, 70%) (hereinafter, the compound of Example 9) was obtained as a colorless oily substance.

[0239] 1 H-NMR (400 MHz, DMSO-d6) δ: 0.94 (6H, t, J=6.8 Hz), 1.05-1.75 (5H, m), 2.42-3.10 (8H, m), 3.64 (3H, s), 3.93-4.02 (1H, m), 4.32-4.43(1H, m), 5.00-5.08(1H, m), 5.34-5.42(1H, m), 6.69-6.71(1H, m), 7.01-7.03(1H, m).

[0240] ESI-MS: m / z=309(M+H) + .

[0241] (Example 10) Evaluation of binding of a cyclic amine derivative (I) to a membrane fraction prepared from each tissue of a rat:

[0242] Regarding the membrane fractions obtained by preparing each tissue of a rat 3 By evaluating the binding ability of the cyclic amine derivative (I) labeled with H, an organization containing the molecular target of the cyclic amine derivative (I) was identified.

[0243] The compound of Example 7 was used as the cyclic amine derivative (I).

[0244] Male SD rats (manufactured by Charles River Laboratories Japan, Inc.) were used at 7 weeks of age. The rats were anesthetized with isoflurane, and euthanasia was induced by hemorrhage from the abdominal aorta using a syringe with an inner wall treated with heparin sodium injection (manufactured by AY Pharmaceuticals Co., Ltd.). Within 60 minutes of euthanasia by hemorrhage, the cerebrum, cerebellum, brainstem, spinal cord, dorsal root ganglion, sciatic nerve, skin, heart, muscle, liver, kidney, and small intestine were excised. For the cerebrum, both cerebral hemispheres were harvested. For the cerebellum, the cerebellum was harvested from the forebrain. For the brainstem, the portion from which the cerebrum and cerebellum had been removed was harvested. The spinal cord was harvested by excising parts of the thoracic and lumbar spinal cords, dissecting the dura mater, and removing the remaining nerve roots. For the dorsal root ganglion, both lumbar ganglia (L3–L6) were harvested. For the sciatic nerve, both sciatic nerves were excised, and attachment tissues such as fat were removed. For the skin, a portion of the skin from the anterior and posterior phalangeal regions was harvested. For the heart, the entire excised portion was harvested. For the muscle, a portion of the quadriceps femoris muscle was harvested. For the liver, the lateral left lobe was harvested. For the kidney, the entire excised portion was harvested. For the small intestine, the contents were cleaned, and approximately 5 cm of the duodenum and perijejunum were harvested. The harvested tissues were immediately frozen with dry ice and preserved at -80°C. The dorsal root ganglion, sciatic nerve, muscle, and skin were mixed from 10 individuals and treated as one sample. For other tissues, tissues from 3 individuals were mixed and treated as one sample.

[0245] Homogenize buffer (sucrose (1M), Tris-HCl (0.5M), and 1× protease inhibitor cocktail (manufactured by Sigma-Aldrich Corporation), pH 7.6) was added to the collected tissues, and homogenate samples of each tissue were prepared using a homogenizer. Each homogenate sample was centrifuged at 1000×g at 4°C for 10 minutes, and undissolved tissue was removed. The supernatant was ultracentrifuged at 10000×g at 4°C for 60 minutes, the resulting precipitate was suspended in Krebs-Ringer bicarbonate buffer (manufactured by Sigma-Aldrich Corporation), and the protein content was quantified using a BCA protein assay kit (manufactured by Thermo Fisher Scientific KK).

[0246] Mechanical division 3 The reaction of the compound of Example 7 labeled with H was carried out entirely in microtubes using the Krebs-Ringer bicarbonate buffer. Sample group A was prepared by adding the compound solution of Example 7 (unlabeled, 400 μm, 100 μL) to the protein solution (0.2 mg / mL, 200 μL) of the membrane fraction of each tissue, and Sample group B was prepared by adding Krebs-Ringer bicarbonate buffer (100 μL).

[0247] For the above samples A and B, containing octyl-β-D-thioglucopyranoside (0.4% (w / v)). 3The H-labeled compound of Example 7 (80 nM, 100 μL) was added, and the mixture was promptly transferred to a 37°C incubator to initiate the reaction. After incubating for 1 hour, the mixture was cooled on ice for at least 5 minutes, and the reaction was stopped. The reaction solution was added to a glass filter (manufactured by Perkinelmer Inc.) that had been pre-immersed in a 0.05% polyethyleneimine solution, and the solution was subjected to suction filtration under rapid depressurization to adsorb the membrane fraction onto the glass filter. After washing the glass filter 6 times with Krebs-Ringer bicarbonate buffer, 10 mL of Hionic-Fluor (manufactured by Perkinelmer Inc.) was added, and the combined 3 For the radioactivity of the compound of Example 7 labeled with H, the breakdown rate (hereinafter referred to as dpm) was measured by the liquid scintillation method. Specific binding (dpm / μg) was calculated as the difference obtained by subtracting the average value of dpm for sample group A from the average value of dpm for sample group B, and dividing this difference by the protein amount (dpm / μg).

[0248] Membrane fractions obtained by preparing each tissue of a rat, and 3 The results of the binding evaluation of the compound of Example 7 labeled with H are shown in FIG. 2. The vertical axis of FIG. 2 represents specific binding (dpm / μg) (average value, 2 examples per group). The horizontal axis represents, from left to right, membrane fractions obtained by preparing each tissue from the cerebrum, cerebellum, brainstem, spinal cord, dorsal root ganglion, sciatic nerve, heart, liver, kidney, small intestine, muscle, and skin.

[0249] As a result, the specific binding of the compound of Example 7 to the membrane fractions prepared from each tissue was significant in the membrane fraction prepared from the dorsal root ganglion. Subsequently, the specific binding of the membrane fraction prepared from the sciatic nerve was 42% of the specific binding of the membrane fraction prepared from the dorsal root ganglion. The specific binding of the membrane fraction prepared from the spinal cord was 23% of the specific binding of the membrane fraction prepared from the dorsal root ganglion. The specific binding of the membrane fraction prepared from the brainstem was 4.2% of the specific binding of the membrane fraction prepared from the dorsal root ganglion, and the specific binding of the membrane fraction prepared from other tissues, including the brain, was less than 1% of the specific binding of the membrane fraction prepared from the dorsal root ganglion. From the fact that the compound of Example 7 specifically binds to a membrane fraction obtained by preparing from a peripheral nerve or spinal cord, such as the dorsal root ganglion or sciatic nerve, it was confirmed that the molecular target of the compound of Example 7 is present in the peripheral nerve or spinal cord, and that the cyclic amine derivative (I) acts by binding to the membrane present in the peripheral nerve or spinal cord.

[0250] (Example 11) Identification of a combined molecule of a cyclic amine derivative (I) or its pharmacologically acceptable salt:

[0251] In order to identify the binding molecule of the cyclic amine derivative (I) or its pharmacologically acceptable salt, affinity purification and comprehensive identification were performed using FG beads immobilized with the cyclic amine derivative (I).

[0252] In order to immobilize FG beads having a cyclic amine derivative (I) and an amino group linker by amide bonding, a cyclic amine derivative having a carboxyl group was synthesized.

[0253] As the above-mentioned cyclic amine derivative having a carboxyl group, (S)-2-(3-(4-(dimethylamino)piperidine-1-yl)-1-(1-methyl-1H-imidazole-2-yl)-3-oxopropoxy)acetic acid represented by the following chemical formula (hereinafter, compound for bead immobilization) was used and synthesized by the following method.

[0254] Synthesis of compounds for bead immobilization:

[0255]

[0256] Palladium carbon (10% wet, 14.0 mg) was added at room temperature to an ethanol (2.50 mL) solution of benzyl(S)-2-(3-(4-(dimethylamino)piperidin-1-yl)-1-(1-methyl-1H-imidazole-2-yl)-3-oxopropoxy)acetate (0.0550 g, 0.128 mmol) and stirred for 6 hours under a hydrogen atmosphere. The reaction mixture was filtered using Cerite, the filtrate was concentrated under reduced pressure, and (S)-2-(3-(4-(dimethylamino)piperidin-1-yl)-1-(1-methyl-1H-imidazole-2-yl)-3-oxopropoxy)acetic acid (0.0404 g, 0.119 mmol, 93%) was obtained as a colorless oily substance.

[0257] 1 H-NMR (400 MHz, CDCl3) δ: 1.30-1.69(2H, m), 1.88-2.10(2H, m), 2.40-2.63(7H, m), 2.88-3.20(3H, m), 3.21-3.41(1H, m), 3.74-3.98(5H, m), 4.09-4.27(1H, m), 4.63-4.78(1H, m), 5.20-5.26(1H, m), 6.84(1H, s), 6.95(1H, s).

[0258] ESI-MS: m / z=339(M+H) + .

[0259] A bead immobilization compound was immobilized on NH2 beads (5 mg, manufactured by TAMAGAWA SEIKI CO., LTD.). First, a bead immobilization compound solution (20 mM, 200 μL), a succinimide solution (200 mM, 20 μL), and a 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide solution (200 mM, 20 μL) were added to N,N-dimethylformamide (160 μL), and the mixture was reacted at room temperature for 2 hours using a microtube mixer to prepare an activated bead immobilization compound solution. Subsequently, N,N-dimethylformamide (900 μL) and the above-mentioned activated bead immobilization compound solution (100 μL) were added to NH2 beads, and the mixture was reacted overnight at room temperature using a microtube mixer. In addition, after washing three times with N,N-dimethylformamide (500 μL), acetic anhydride was added to make the concentration 20%, and the mixture was reacted at room temperature for 2 hours using a microtube mixer. Finally, the mixture was washed three times with 50% methanol (500 μL) to form cyclic amine derivative immobilized beads.

[0260] The protein solution reacted with the above-mentioned cyclic amine derivative immobilized beads was prepared from rat dorsal root ganglia. The rat dorsal root ganglia were isolated from 6-week-old male SD rats (manufactured by Charles River Laboratories Japan, Inc.). The rats were anesthetized with urethane, and euthanized by hemorrhage after incising the abdominal aorta. After incising the posterior dorsal region, the vertebral column was extracted and ice-chilled. The dorsal side of the vertebral column was cut, and after removing the spinal cord from the ventral side of the vertebral column, the dorsal root ganglia (T11 to 13 and L3 to 6, bilateral) were extracted along the nerve fiber bundles using precision forceps. The extracted dorsal root ganglia were immersed in ice-chilled Leibovitz's L-15 medium (manufactured by Thermo Fisher Scientific KK), and the nerve fiber bundles were removed and the dorsal root ganglia were isolated under a stereomicroscope. The isolated dorsal root ganglion was frozen with liquid nitrogen and stored at -80°C. The dorsal root ganglion was mixed with tissues taken from 21 individuals and treated as one sample.

[0261] The dorsal root ganglion was homogenized using a Downs-type homogenizer in assay buffer (NaCl (140 mM), KCl (5 mM), CaCl2·2H2O (1.2 mM), MgCl2·6H2O (2 mM), D(+)-glucose (14 mM), Hepes (10 mM), and protease inhibitor cocktail (manufactured by Sigma-Aldrich Corporation), pH 7.4) to prepare a dorsal root ganglion lysate. The surfactant n-Dodecyl-β-D-Maltopyranoside (manufactured by Sigma-Aldrich Corporation) was added to the dorsal root ganglion lysate to a concentration of 1%, and the mixture was incubated at 4°C for 1 hour. The mixture was centrifuged at 20,400×g for 30 minutes, and the supernatant was diluted 40-fold and concentrated with Amicon Ultra 3kDa (manufactured by Merck Millipore).

[0262] The cyclic amine derivative immobilized beads were mixed with the protein solution and reacted overnight at 4°C. The beads were washed three times with assay buffer, washed once with PBS(-), and then suspended in PBS(-). The proteins bound to the beads were comprehensively identified by shotgun analysis (LC-MS / MS).

[0263] Proteins that do not bind to beads that do not immobilize compounds but bind to beads that immobilize cyclic amine derivatives are shown in Table 3. Entry IDs (rats) and protein names were cited from the protein database Uniprot (http: / / www.uniprot.org / ). However, regarding the protein name of entry ID F1LTJ5 in Table 3, although it was Uncharacterized Protein in Uniprot, it was an ortholog of basement membrane type heparan sulfate proteoglycan core protein in the PANTHER Classification System, so the protein name was given as basement membrane type heparan sulfate proteoglycan core protein.

[0264]

[0265] Among the eight identified molecules, adbilin has been reported to be highly expressed in peripheral nerve tissue compared to other tissues (Ravena II et al., European Journal of Neuroscience, 2002, Vol. 27, pp. 14404-14414). In Example 10, it was suggested that the molecular target of the compound of Example 7 is adbilin, based on the fact that the molecular target of the cyclic amine derivative (I) is a molecule that is abundant in peripheral nerves.

[0266] In addition, it has been reported that adbilin forms a complex with myosin, etc., identified in this embodiment (Chuang et al., Proceedings of the National Academy of Sciences of the United States of America, 2018, Vol. 115, pp. E 8557-E8566). Thus, it has been suggested that a cyclic amine derivative (I) or a pharmacologically acceptable salt thereof may bind to an adbilin complex composed of adbilin and / or the protein, etc., identified in this embodiment, and modulate the function of adbilin.

[0267] (Example 12) Intracellular actin filament reduction effect in rat dorsal root ganglion neuron cell lines:

[0268] The effect of cyclic amine derivatives (I) or their pharmacologically acceptable salts on the amount of intracellular actin filaments in F11 cells (rat dorsal root ganglion neurite cells) was examined.

[0269] Compounds of Examples 1, 2, 3, 4, 5, 6, 7, 8, and 9 were used as test substances.

[0270] Compounds of the comparative examples include those shown in Table 4: 1-(4-(dimethylamino)piperidin-1-yl)-3-(1-ethyl-1H-imidazole-2-yl)-3-hydroxypropan-1-one (hereinafter, compound of Comparative Example 1), 1-(4-(dimethylamino)piperidin-1-yl)-3-hydroxy-3-(1-(2,2,2-trifluoroethyl)-1H-imidazole-2-yl)propan-1-one (hereinafter, compound of Comparative Example 2), 1-((R)-3-(dimethylamino)pyrrolidin-1-yl)-3-hydroxy-3-(1-methyl-1H-imidazole-2-yl)propan-1-one (hereinafter, compound of Comparative Example 3), and 1-((R)-3-(3-(dimethylamino)piperidine-1-yl)-3-hydroxy-3-(1-methyl-1H-imidazole-2-yl)propan-1-one (hereinafter, the compound of Comparative Example 4) was used. The compounds of Comparative Examples 1, 2, 3 and 4 were synthesized according to the method described in the known literature (International Publication No. 2016 / 136944).

[0271]

[0272] To detach F11 cells from the culture flask, 0.25% trypsin / EDTA (manufactured by Thermo Fisher Scientific KK) was added to the cells, and they were incubated in water at room temperature. Subsequently, DMEM medium containing 10% fetal bovine serum (manufactured by Thermo Fisher Scientific KK) was added to suspend the cells, and the mixture was centrifuged at 1,500 rpm for 1 minute. After removing the supernatant, DMEM medium containing 0.5% fetal bovine serum was added to suspend the cells, and a laminin coat (2 μg / cm²) was applied. 2The cells were seeded into a poly-D-lysine coated 96-well plate (manufactured by BD Bioscience) and cultured at 37°C under a 5% CO2 environment. Two hours after seeding, N,N-dibutyladenosine 3',5'-phosphate (final concentration 1 mM, manufactured by Sigma-Aldrich Corporation) and recombinant rat GDNF (final concentration 50 ng / mL, manufactured by PeproTech Inc.) were added, and differentiation was induced by incubating for 7 days at 37°C under a 5% CO2 environment.

[0273] After 7 days of culture initiation, neuronal axonal damage was induced by switching to DMEM medium containing 0.5% fetal bovine serum containing oxaliplatin (final concentration 15 μM) and culturing for 5 days. Each compound of the Examples or Comparative Examples was treated by including it in the medium along with oxaliplatin (final concentration 10 μM).

[0274] After 5 days of treatment with oxaliplatin and each compound, cells were washed three times with PBS(-), and fixed by adding 4% paraformaldehyde-phosphate buffer (manufactured by Wako Pure Chemical Co., Ltd.) and incubating at room temperature for 10 minutes. Actin filaments were stained by adding Acti-stain 670 phalloidin (manufactured by Cytoskeleton) and incubating at room temperature for 30 minutes. Cell images were acquired using an IN Cell Analyzer 2200 (manufactured by GE Healthcare), and image analysis was performed using the IN Cell Investigator Developer Toolbox. The amount of intracellular actin filaments was expressed as the fluorescence intensity (density) of the fluorescently labeled phalloidin bound to the actin filaments.

[0275] It was confirmed that the group treated with oxaliplatin alone (hereinafter, the oxaliplatin alone treatment group) showed a 5% increase in intracellular actin filament volume compared to the group not treated with oxaliplatin, and that it caused abnormal regulation of actin filament metabolic turnover (p<0.05, Student's t-test). The reduction rate of intracellular actin filaments by each compound was calculated for the intracellular actin filament volume of this oxaliplatin alone treatment group.

[0276] The effect of each compound on reducing intracellular actin filaments in F11 cells is shown in Table 5. The “intracellular actin filament reduction rate” in the table represents the ratio (%) of the amount of intracellular actin filaments reduced by treatment with each compound relative to the amount of intracellular actin filaments in the group treated with oxaliplatin alone (average value, the number of examples for each group is 5 as one example (1 well)). The “subject compound” in the table represents the compound treatment group for each compound.

[0277]

[0278] Compared to the solvent treatment group, the compounds of Examples 1, 2, 3, 4, 5, 6, 7, 8, and 9 reduced the amount of actin filaments in the cells by 5% or more. Meanwhile, the reduction rate of the amount of actin filaments in the cells for the compounds of Comparative Examples 1, 2, 3, and 4 was 5% or less.

[0279] (Example 13) Action to improve abnormal actin filament metabolic rotation regulation in a rat spinal cord ligation model:

[0280] The action of cyclic amine derivatives (I) or pharmacologically acceptable salts thereof on the ratio of actin filament amount to monomeric actin amount in the sciatic nerve of a rat spinal nerve ligation model was examined.

[0281] Male SD rats (manufactured by Charles River Laboratories Japan, Inc.) were used at 6 weeks of age. The rat spinal nerve ligation model was constructed by incising the right lumborum of the rat under isoflurane inhalation anesthesia, resecting the spinal laminae, completely ligating the L5 and L6 nerve roots of the spinal nerves, and suturing the muscle layer and skin. For the negative control (simulated surgery group), the right L5 and L6 nerves were exposed, and the muscle layer and skin were sutured without ligation. Rats one week after nerve ligation were used in the following experiments.

[0282] In a rat spinal cord ligation model, the compound of Example 7 was orally administered once daily for 2 days at three doses of 6, 20, or 60 mg / kg. Three hours after the final administration, the sciatic nerve was harvested under isoflurane anesthesia, and the rats were euthanized. The harvested sciatic nerves were cryopreserved at -80°C. As controls, a simulated surgery group administered distilled water as a solvent and a distilled water administration group of the spinal cord ligation model (distilled water administration group) were established. In each case, the sciatic nerves were harvested by the same treatment as above, and the rats were euthanized.

[0283] A G-actin / F-actin in vivo assay kit (manufactured by Cytoskeleton) was used for the quantification of actin filaments and monomeric actin. Frozen tissues were homogenized by adding lysis and F-actin stabilization buffer (manufactured by Cytoskeleton) containing a protease inhibitor and ATP (1 mM). The homogenate was incubated at 37°C for 10 minutes and centrifuged at 350×g for 5 minutes. The supernatant was centrifuged at 100,000×g at 37°C for 1 hour. The supernatant obtained after centrifugation was designated as the monomeric actin sample, and the precipitate as the actin filament sample. The actin filament sample was incubated on ice for 1 hour and pipetted once every 15 minutes. The amount of actin in the monomeric actin sample and the actin filament sample was measured by Western blot. Monomeric actin samples and actin filament samples were heat-denatured by adding SDS sample buffer, applied to an SDS-PAGE precast (manufactured by BIO-RAD), and subjected to electrophoresis to separate proteins. Proteins in the gel were electrically transferred and immobilized onto a PVDF membrane to produce a blot (blot membrane) (Trans Blot Turbo Blotting System, manufactured by BIO-RAD). After blocking the blot membrane (Block One, manufactured by NACALAI TESQUE, INC.), an antibody against actin (anti-actin rabbit polyclonal antibody) was added, and the sample was incubated at room temperature for 1 hour. After washing the blot membrane, an anti-rabbit HRP-labeled secondary antibody (manufactured by GE Healthcare) was added, and the sample was incubated at room temperature for 30 minutes. After washing the blot membrane, a detection reagent (ECL Prime, manufactured by GE Healthcare) was added, and chemiluminescence was detected using a CCD imager (manufactured by Bio-RAD). The luminescence intensity of the actin band was quantified using Image Lab software (manufactured by Bio-RAD).

[0284] Figure 3 shows the results of evaluating the actin filament / monomer actin ratio in the sciatic nerve of a rat spinal cord ligation model using the compound of Example 7. Figure 3a shows images of actin proteins detected by Western blot in an actin filament sample (top) and a monomer actin sample (bottom) isolated from the sciatic nerve. The luminescence intensity of the bands of each actin protein was quantified, and the ratio of actin filaments to monomer actin (hereinafter, the actin filament / monomer actin ratio) was calculated and is shown in Figure 3b. The vertical axis of Figure 3b represents the actin filament / monomer actin ratio (mean value ± standard error). On the horizontal axis, "distilled water simulated surgery" represents the simulated surgery group (6 cases), "distilled water" represents the distilled water administration group (7 cases) of the spinal cord ligation model, "compound of Example 7" represents the administration group of the compound of Example 7 of the spinal cord ligation model, "6" represents the 6 mg / kg administration group (6 cases) of the compound of Example 7 of the spinal cord ligation model, "20" represents the 20 mg / kg administration group (6 cases) of the compound of Example 7 of the spinal cord ligation model, and "60" represents the 60 mg / kg administration group (7 cases) of the compound of Example 7 of the spinal cord ligation model. "#" in the diagram indicates a statistically significant difference (#: p < 0.05, Student's t-test) compared to the simulated surgery group, and "*" in the diagram indicates a statistically significant difference (*: p < 0.025, Williams' multiple comparison, unilateral) compared to the distilled water administration group of the spinal cord ligation model.

[0285] In the rat spinal cord ligation model group administered distilled water (distilled water administration group), the actin filament / monomer actin ratio was significantly increased compared to the simulated surgery group. In the group administered the compound of Example 7, the actin filament / monomer actin ratio decreased with increasing dosage of the compound of Example 7, and a significant decrease was confirmed in the 60 mg / kg administration group compared to the distilled water administration group. In other words, it was clearly established that the compound of Example 7 has the effect of improving the abnormal regulation of actin filament metabolic turnover of the sciatic nerve caused by spinal cord ligation.

[0286] It has been reported that one of the molecular targets of the cyclic amine derivative (I) or its pharmacologically acceptable salt shown in Example 11, adbilin, has the function of regulating actin filament metabolic turnover through binding to actin (Hasegawa et al., The Journal of Neuroscience, 2007, Vol. 27, pp. 14404-14414). Accordingly, from Examples 11, 12, and 13, it became evident that the cyclic amine derivative (I) or its pharmacologically acceptable salt promotes the function of adbilin and / or the adbilin complex, along with binding to adbilin and / or the adbilin complex, and exhibits an improved effect beyond the regulation of actin filament metabolic turnover.

[0287] (Example 14) Evaluation of nerve axon extension-promoting action in rat dorsal root ganglion primary culture cells:

[0288] The action of cyclic amine derivatives (I) or pharmacologically acceptable salts thereof on nerve process extension in rat dorsal root ganglion primary culture cells was examined.

[0289] Male SD rats (manufactured by Charles River Laboratories Japan, Inc.) aged 4 to 7 weeks were used. The rats were anesthetized by intraperitoneal administration of urethane. After confirming that anesthesia had been achieved, the abdominal aorta was promptly incised, and the rats were euthanized by hemorrhage. After incising the posterior dorsal region, the vertebral column was excised and ice-cold. The dorsal side of the vertebral column was cut, the spinal cord was removed from the ventral side, and the ventral dorsal root ganglion was exposed. The dorsal root ganglion (L3 to L6, bilateral) was excised with forceps, immersed in ice-cold Leibovitz's L-15 medium (manufactured by Thermo Fisher Scientific KK), and the nerve fiber bundles were removed under a stereomicroscope. The dorsal root ganglion from which the nerve fiber bundles were removed was finely incised with ophthalmic scissors to create a dorsal root segment.

[0290] The dorsal root segment was immersed in approximately 3 mL of collagenase A solution (manufactured by Roche Molecular Systems Inc.) and incubated at 37°C for 20 minutes. After incubation, the supernatant collagenase A solution was removed by centrifugation at 200×g at 25°C for 5 minutes, approximately 1 mL of trypsin solution was added, and the mixture was incubated at 37°C for 5 minutes. Subsequently, approximately 3 mL of DMEM containing 10% fetal bovine serum (manufactured by Thermo Fisher Scientific KK) was added, and the supernatant was removed after centrifugation at 200×g at room temperature for 5 minutes. After removing the supernatant, about 5 mL of Neurobasal-A Medium containing 2% B27 supplement (hereinafter referred to as culture medium, manufactured by Thermo Fisher Scientific KK) was added to the precipitate and dispersed using a micropipette. Then, the cells from which debris had been removed were recovered using a 70 µm cell strainer (manufactured by BD Falcon), and the cells were recovered by washing the debris on the cell strainer in 5 mL of culture medium. The recovered cells were centrifuged at 200×g at room temperature for 5 minutes, and the supernatant was removed.

[0291] After removing the supernatant, 5 mL of culture medium was added, and the cell pellet was suspended. This cell suspension was coated with a laminin coat (6 μg / cm²). 2 100 μL of poly-D-lysine was seeded into 96-well plates (manufactured by BD Bioscience) and cultured in a CO2 incubator at 37°C and 5% CO2.

[0292] After 2 hours of incubation, the culture medium was replaced with one containing oxaliplatin (final concentration 3 μM), and neuronal axonal damage was induced by incubating for 7 days. The compound of Example 7 was added to the culture medium simultaneously with the oxaliplatin treatment (final concentrations 10, 30, or 100 μM).

[0293] One week after the start of culture, the cells were washed three times with PBS(-) and treated with 4% paraformaldehyde-phosphate buffer for 10 minutes. After washing three times with PBS(-), the cells were treated with 20% Blocking One (manufactured by NACALAI TESQUE, INC.) and 0.1% Trypton X-100 / PBS(-) for 1 hour. After washing once with PBS(-), the anti-βIII tube-in mouse monoclonal antibody (manufactured by Promega Corporation) was diluted 500-fold with 0.1% Trypton X-100 / PBS(-) containing 5% Blocking One and treated overnight at 4°C. After overnight treatment, the sample was washed three times with PBS(-), and a secondary antibody (CF488A anti-mouse IgG donkey antibody, manufactured by Biotium) was diluted with 0.1% Trypton X-100 / PBS(-) containing 5% blocking agent and treated at room temperature for 1 hour. Subsequently, after washing three times with PBS(-), a DAPI (manufactured by Dojindo Molecular Technologies, Inc.) diluted 1000-fold with PBS(-) was treated for 5 minutes and washed three times with PBS(-).

[0294] Cell images were acquired using the IN Cell Analyzer 2200 (manufactured by GE Healthcare), and image analysis was performed using the IN Cell Investigator Developer Toolbox. The analysis results were expressed as the length of the neurite per cell by dividing the fiber length of the region recognized as a neurite by the number of nerve cells, and the effect of increasing the length of the neurite was evaluated as a nerve axon stretching effect.

[0295] Figure 4 shows images of average cell examples under each condition: an untreated group (treated with distilled water), a group treated with oxaliplatin alone, and a group treated with 100 μM of oxaliplatin and the compound of Example 7. As shown in Figure 4, a shortening of the neurite length was observed with treatment with oxaliplatin alone, and an increase in the neurite length was confirmed with treatment with the compound of Example 7 (100 μM) compared to the group treated with oxaliplatin alone.

[0296] The results of the analysis of the length of the nerve processes in each image are shown in Fig. 5. The vertical axis of Fig. 5 represents the length of the nerve processes (μm / nerve cell) (mean value ± standard error, 5 examples per group). On the horizontal axis, "distilled water" represents the untreated group, "oxaliplatin + distilled water" represents the oxaliplatin-only treatment group, "oxaliplatin + compound of Example 7" represents the group administered with oxaliplatin and the compound of Example 7, "10" represents the 10 μM treatment group of oxaliplatin and the compound of Example 7, "30" represents the 30 μM treatment group of oxaliplatin and the compound of Example 7, and "100" represents the 100 μM treatment group of oxaliplatin and the compound of Example 7. "#" in the diagram indicates a statistically significant difference compared to the no-treatment group (#: p < 0.05, Student's t-test), and "*" in the diagram indicates a statistically significant difference compared to the oxaliplatin-only treatment group (*: p < 0.025, Williams' multiple comparison, unilateral).

[0297] As shown in Figure 5, the length of the neurite was significantly shortened by treatment with oxaliplatin alone, but treatment with the compound of Example 7 (10, 30, and 100 μM) confirmed an increase in the length of the neurite with increasing dose of the compound of Example 7 compared to the group treated with oxaliplatin alone. In the group treated with 100 μM of the compound of Example 7, a significant increase was confirmed compared to the group treated with oxaliplatin alone. That is, it was confirmed that the compound of Example 7 has a neuroaxon extension-promoting effect in rat dorsal root ganglion primary culture cells in which neuroaxon damage was induced with oxaliplatin.

[0298] It has been reported that the function of adbilin, a molecular target of the cyclic amine derivative (I) shown in Example 11, is to stretch nerve axons (Hasegawa et al., The Journal of Neuroscience, 2007, Vol. 27, pp. 14404-14414). Accordingly, from Example 11 and Example 14, it has been shown that the cyclic amine derivative (I) or its pharmacologically acceptable salt has a nerve axon stretching promoting effect in addition to binding to adbilin and / or adbilin complexes.

[0299] From the above results, it has become clear that the cyclic amine derivative (I) of the claims of the present invention has an adbilin function-promoting effect.

[0300] Industrial applicability

[0301] The cyclic amine derivative of the present invention or its pharmacologically acceptable salt can be used as a medicine for diseases related to nerve axonal damage, due to the fact that it has an adbilin function-promoting effect.

[0302] All publications, patents, and patent applications cited in this specification are incorporated herein by reference.

Claims

Claim 1 A pharmaceutical composition for treating nerve axonal damage, comprising as an active ingredient a cyclic amine derivative represented by the following general formula (I) or a pharmacologically acceptable salt thereof, wherein the nerve axonal damage is nerve axonal damage caused by a disease selected from the group consisting of giant cell vasculitis, Takayasu's arteritis, nodular polyarteritis, Kawasaki disease, polyangiitis granuloma, microscopic polyangiitis, eosinophilic polyangiitis granulomatosis, clogioglobulinemia, IgA vasculitis, cutaneous leukocytic vasculitis, systemic erythematosis, Sjögren's syndrome, rheumatoid arthritis, mixed connective tissue disease, polymyositis, dermatomyositis, scleroderma, Behcet's disease, bacterial or viral infection, sarcoidosis, and malignant tumor. [In the formula, carbons marked with * are non-carbons, and A represents a group represented by the following general formula (IIa) or (IIb), and R 1 represents a methyl group, n-propyl group, isopropyl group, 2-methoxyethyl group, 2-ethoxyethyl group, difluoromethyl group, or 3,3,3-trifluoropropyl group, and R 2 represents a hydrogen atom, fluorine atom, or chlorine atom, and R 3 Each independently represents a methyl or ethyl group, and X is -O- or -N(R 3 )- represents] Claim 2 In claim 1, A is a medicinal composition represented by the general formula (IIa). Claim 3 In claim 1, A is a medicinal composition represented by the general formula (IIb). Claim 4 In the third clause, X is -N(R 3 )-Pharmaceutical composition. Claim 5 In any one of paragraphs 1 to 4, R 1 A pharmaceutical composition comprising a methyl group, an n-propyl group, an isopropyl group, a 2-methoxyethyl group, or a 3,3,3-trifluoropropyl group. Claim 6 In any one of paragraphs 1 to 4, R 2 A pharmaceutical composition that is a hydrogen atom or a chlorine atom. Claim 7 A pharmaceutical composition according to any one of claims 1 to 4, wherein the stereochemistry of the carbon with * attached to the subtitle is the S configuration. Claim 8 A pharmaceutical composition for promoting the function of Advilin by binding to Advilin and / or an Advilin complex, in any one of claims 1 to 4. Claim 9 A pharmaceutical composition for improving abnormal actin filament metabolic turnover regulation in any one of claims 1 to 4. Claim 10 A pharmaceutical composition for promoting nerve axon extension in any one of claims 1 to 4. Claim 11 A pharmaceutical composition according to any one of claims 1 to 4, for improving abnormal regulation of actin filament metabolic turnover and promoting neuronal axon extension by binding to Advilin and / or an Advilin complex. Claim 12 A pharmaceutical composition according to claim 1, wherein the bacterial or viral infection is Lyme disease, HIV infection, or Hansen's disease. Claim 13 3-hydroxy-3-(1-methyl-1H-imidazole-2-yl)-1-(4-morpholinopiperidin-1-yl)propan-1-one, 1-(4-(dimethylamino)piperidin-1-yl)-3-(1-propyl-1H-imidazole-2-yl)-3-hydroxypropan-1-one, 1-(4-(dimethylamino)piperidin-1-yl)-3-(1-isopropyl-1H-imidazole-2-yl)-3-hydroxypropan-1-one, 3-(5-chloro-1-methyl-1H-imidazole-2-yl)-1-(4-(dimethylamino)piperidin-1-yl)-3-hydroxypropan-1-one, A cyclic amine derivative or a pharmacologically acceptable salt thereof, which is a compound selected from the group consisting of 1-(4-(dimethylamino)piperidin-1-yl)-3-(1-(2-methoxyethyl)-1H-imidazole-2-yl)-3-hydroxypropan-1-one and 1-(4-(dimethylamino)piperidin-1-yl)-3-(1-(3,3,3-trifluoropropyl)-1H-imidazole-2-yl)-3-hydroxypropan-1-one. Claim 14 A pharmaceutical composition for the treatment of neuronal axonal damage, comprising as an active ingredient a cyclic amine derivative or a pharmacologically acceptable salt thereof as described in claim 13, wherein the neuronal axonal damage is neuronal axonal damage caused by a disease selected from the group consisting of giant cell vasculitis, Takayasu's arteritis, nodular polyarteritis, Kawasaki disease, polyangiitis granuloma, microscopic polyangiitis, eosinophilic polyangiitis granulomatosis, clogioglobulinemia, IgA vasculitis, cutaneous leukocytic vasculitis, systemic erythematosis, Sjögren's syndrome, rheumatoid arthritis, mixed connective tissue disease, polymyositis, dermatomyositis, scleroderma, Behcet's disease, bacterial or viral infection, sarcoidosis, and malignant tumor. Claim 15 In claim 14, the above bacterial or viral infection is Lyme disease, HIV infection, or Hansen's disease, in a pharmaceutical composition.

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