JP2024149429A - Metal transporter ZIP14 inhibitor - Google Patents

Abstract

The present invention provides a compound that has an inhibitory effect on the metal transporter ZIP14 and can be used as a preventive or therapeutic agent for diseases associated with the metal transporter ZIP14. The present invention relates to a compound represented by formula (I): JPEG2024149429000025.jpg5752. [Selected Figure] Figure 1

Description

The present invention relates to a metal transporter ZIP14 inhibitor, and more particularly to a metal transporter ZIP14 inhibitor that does not inhibit the metal transporter ZIP8.

Zinc, iron, and manganese are essential metals (essential trace metals) for the maintenance of life. While essential trace metals are required for normal cell function, their excessive accumulation in cells causes cell damage. The homeostasis of essential trace metals inside and outside the cells is strictly controlled by metal transporters, which contribute to the maintenance of life by regulating the amount and distribution of metals in the body (Non-Patent Document 1).
Metal transporter ZIP14 (gene name: Slc39al4) (hereinafter, simply referred to as ZIP14) transports essential trace metals such as zinc, iron, and manganese, as well as the toxic metal cadmium (Non-Patent Document 2). In recent years, it has been shown that excessive intracellular accumulation of metals by ZIP14 is involved in various diseases. Specifically, ZIP14 expressed in skeletal muscle due to the action of inflammatory cytokines impairs the differentiation and maintenance of skeletal muscle by introducing excessive zinc ions into skeletal muscle cells, and is involved in the development of cancer cachexia accompanied by atrophy and loss of skeletal muscle (Non-Patent Document 3). Furthermore, ZIP14 expressed in the liver promotes liver damage associated with iron overload (Non-Patent Document 4). In addition, ZIP14 imbalance leads to cytotoxicity due to the accumulation of manganese and cadmium (Non-Patent Documents 5 and 6).
These findings indicate that dysfunction of ZIP14 disrupts metal homeostasis in the body and damages cells that compose tissues such as skeletal muscle and liver, suggesting that ZIP14 is a disease-related molecule and a drug discovery target molecule for the target disease (Non-Patent Document 7).
In addition, because ZIP14 expression is increased by the action of inflammatory cytokines (Non-Patent Document 3), disorders caused by excessive metal transport by ZIP14 may have a ripple effect on abnormalities in various organs associated with acute and chronic inflammation.
Furthermore, Non-Patent Document 10 suggests that ZIP14 is involved in the control of intracellular zinc homeostasis in cancer cells derived from liver cancer and may play a major role in controlling the intracellular zinc deficiency observed in these cancer cells.

From the above, it is suggested that ZIP14 inhibitors may serve as preventive or therapeutic agents for diseases associated with ZIP14 (e.g., cancer cachexia accompanied by skeletal muscle atrophy or loss, iron overload (including liver damage associated with iron overload), liver cancer, manganese or cadmium poisoning, etc., particularly cancer cachexia accompanied by skeletal muscle atrophy or loss).
ZIP8 is known as the metal transporter that is most closely related to ZIP14 in terms of molecular evolution (Non-Patent Document 2).

On the other hand, Patent Documents 1 to 8 disclose that compounds having a 1,3,8-triazaspiro[4.5]decan-4-one skeleton are useful for treating diseases related to opioid receptors/ORL-1 receptors. Non-Patent Document 8 discloses that compounds having a 1,3,8-triazaspiro[4.5]decan-4-one skeleton have affinity and selectivity for the mu receptor. Non-Patent Document 9 discloses that compounds having a 1,3,8-triazaspiro[4.5]decan-4-one skeleton have inhibitory activity against glycine transporter 1 (GlyT1). However, none of the documents teach any relationship with ZIP14.

WO 02/083673 A1 WO 2001/039723 A3 WO 01/07050 A1 WO 01/36418 A1 WO 00/06545 A1 WO 99/59997 A1 EP 0997464 A1 EP 0856514 A1

Metallomics 3:535-750, 2011 Biometals 25:643-55, 2012 Nature Medicine 24:770-781, 2018 Cell Metabolism 22:138-150, 2015 Proc. Natl. Acad. Sci. USAJ 15: E1769-E1778, 2018 Toxicological Sciences 143: 26-35, 2015 J. Pharmacol. Sci. 148:221-228, 2022 Mol Pharmacol. 41:185-96, 1992 Bioorganic & Medicinal Chemistry Letters 16: 349-353, 2006 PeerJ 9:e12314, 2021 Ministry of Health, Labor and Welfare, 13th Pharmaceuticals and Medical Devices Administration Evaluation and Monitoring Committee Document 4 "Overseas Surveys by the Pharmaceuticals and Medical Devices Administration Evaluation and Monitoring Committee"

As a symptom of cancer cachexia, a sustained decrease in skeletal muscle mass is observed, palliative treatment such as nutritional support is currently being used. In recent years, a ghrelin receptor agonist (anamorelin), which is expected to increase appetite, has been approved in Japan for the treatment of cancer cachexia with poor prognosis. However, as of September 20, 2023, the European Medicines Agency and the US Food and Drug Administration have not approved anamorelin for the treatment of cancer cachexia, stating that the efficacy of anamorelin has not been sufficiently proven (Non-Patent Document 11). In other words, there is currently no cure for cancer cachexia that acts directly on skeletal muscle, and effective therapeutic supplements are needed.

Therefore, there is a strong need for the development of a drug for treating cancer cachexia that can improve the loss of skeletal muscle mass. There is also a need for the development of a drug for treating iron overload (including liver damage associated with iron overload), liver cancer, or manganese or cadmium poisoning.

The object of the present invention is to provide a compound that has an inhibitory effect on the metal transporter ZIP14 and can be used as a preventive or therapeutic agent for diseases associated with the metal transporter ZIP14 (e.g., cancer cachexia accompanied by skeletal muscle atrophy or loss, iron overload (including liver damage associated with iron overload), liver cancer, manganese or cadmium poisoning, etc., particularly cancer cachexia accompanied by skeletal muscle atrophy or loss).

As a result of intensive research aimed at solving the above problems, the present inventors have found that the compound represented by the following formula (I) has an inhibitory effect on the metal transporter ZIP14 and is therefore useful as a preventive or therapeutic agent for diseases related to the metal transporter ZIP14 (e.g., cancer cachexia accompanied by skeletal muscle atrophy or loss, iron overload (including liver damage associated with iron overload), liver cancer, manganese or cadmium poisoning, etc., particularly cancer cachexia accompanied by skeletal muscle atrophy or loss), and have completed the present invention.

That is, the present invention is as follows.
[1] Formula (I):

[Wherein,
R ¹ represents a C _6-10 aryl group, a C _7-14 aralkyl group, a 5- or 6-membered aromatic heterocyclic group, a 3- to 8-membered non-aromatic heterocyclic group, a C _3-8 cycloalkyl group, a C _3-8 cycloalkenyl group, or a C _1-6 alkyl group, in which the C _6-10 aryl group, C _7-14 aralkyl group, a 5- or 6-membered aromatic heterocyclic group, a 3- to 8-membered non-aromatic heterocyclic group, a C _3-8 cycloalkyl group, or a C _3-8 cycloalkenyl group is optionally substituted with a substituent selected from the substituent group Z;
R ² represents a hydrogen atom or a C _1-6 alkyl group;
R ³ represents a hydrogen atom, a C _1-6 alkyl group, a C _6-10 aryl group, a C _7-14 aralkyl group, a 5- or 6-membered aromatic heterocyclic group, a 3- to 8-membered non-aromatic heterocyclic group, a C _3-8 cycloalkyl group, or a C _3-8 cycloalkenyl group, in which the C _6-10 aryl group, C _7-14 aralkyl group, a 5- or 6-membered aromatic heterocyclic group, a 3- to 8-membered non-aromatic heterocyclic group, a C _3-8 cycloalkyl group, or a C _3-8 cycloalkenyl group is optionally substituted by a substituent selected from the substituent group Z;
R ⁴ is selected from a halogen atom, a cyano group, a nitro group, a C _1-6 alkyl group, a C _1-6 haloalkyl group, a hydroxy group, a C _1-6 alkoxy group, a C _1-6 haloalkoxy group, an amino group, a mono-C _1-6 alkylamino group, a di-C _1-6 alkylamino group, a carboxy group, a C _1-6 alkyl-carbonyl group, and a C _1-6 alkoxy-carbonyl group;
n represents an integer of 0 to 2;
^R5 is a hydrogen atom, a _C1-6 alkyl group, or a group represented by the formula: -L-Cy
(Wherein,
L is a bond, a C _1-6 alkylene group, or a group of the formula: *-(CH ₂ ) _m1 -X-(CH ₂ ) _m2 -**, where:
X represents O, S, NR ⁶ , CO, NR ⁶ CO, CONR ⁶ , COO, or OCO, R ⁶ represents a hydrogen atom or a C _1-6 alkyl group;
m1 represents an integer of 0 to 3;
m2 represents an integer of 0 to 3;
* indicates the bonding site with the nitrogen atom of the spiro ring.
** indicates the binding site with Cy.)
represents a group represented by
Cy represents a C _6-10 aryl group, a 5- or 6-membered aromatic heterocyclic group, a 3- to 8-membered non-aromatic heterocyclic group, a C _3-8 cycloalkyl group, or a C _3-8 cycloalkenyl group, in which the C _6-10 aryl group, the 5- or 6-membered aromatic heterocyclic group, the 3- to 8-membered non-aromatic heterocyclic group, the C _3-8 cycloalkyl group, or the C _3-8 cycloalkenyl group may be substituted with a substituent selected from the substituent group Z.
represents a group represented by
Substituent group Z consists of a halogen atom, a cyano group, a nitro group, a C _1-6 alkyl group, a C _1-6 haloalkyl group, a hydroxy group, a C _1-6 alkoxy group, a C _1-6 haloalkoxy group, an amino group, a mono-C _1-6 alkylamino group, a di-C _1-6 alkylamino group, a carboxy group, a C _1-6 alkyl-carbonyl group, a C _1-6 alkoxy-carbonyl group, an azido group, a carbamoyl group, a mono-C _1-6 alkyl-carbamoyl group, a di-C _1-6 alkyl-carbamoyl group, a sulfamoyl group, a mono-C _{1-6 alkyl-sulfamoyl group, a di-C 1-6 alkyl} -sulfamoyl group, a sulfo group, a methylenedioxy group (-OCH ₂ O-), and a C _6-10 aryl group.]
or a salt thereof (hereinafter also referred to as compound (I)).

[2] The inhibitor according to the above-mentioned [1], wherein R ¹ is a C _6-10 aryl group, a C _7-14 aralkyl group, a 5- or 6-membered aromatic heterocyclic group, a 3-membered to 8-membered non-aromatic heterocyclic group, a C _3-8 cycloalkyl group, or a C _3-8 cycloalkenyl group, wherein the C _6-10 aryl group, a C _7-14 aralkyl group, a 5- or 6-membered aromatic heterocyclic group, a 3-membered to 8-membered non-aromatic heterocyclic group, a C _3-8 cycloalkyl group, or a C _3-8 cycloalkenyl group is optionally substituted by a substituent selected from substituent group Z.
[3] The inhibitor according to the above-mentioned [1], wherein R ² is a hydrogen atom.
[4] The inhibitor according to the above-mentioned [1], wherein R ³ is a hydrogen atom or a C _1-2 alkyl group.
[5] The inhibitor according to the above-mentioned [1], wherein n is 0.
[6] R ⁵ is a group represented by the formula: -L-Cy;
L is a C _1-6 alkylene group; and
Cy has the same meaning as [1] above.
The inhibitor described in [1] above.
[7] A compound represented by formula (I) or a salt thereof,
1-phenyl-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 3) or a salt thereof;
The inhibitor according to [1], which is 8-(2-(2-azidophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 37) or a salt thereof; or 1-phenyl-8-(2-(4-hydroxy-2-methylphenyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 39) or a salt thereof.
[8] The inhibitor according to the above-mentioned [1], wherein the compound represented by the formula (I) or a salt thereof is 1-phenyl-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one or a hydrochloride thereof.

[9] The inhibitor according to the above-mentioned [1], which inhibits transport of zinc, iron, manganese or cadmium via the metal transporter ZIP14.
[10] The inhibitor according to the above-mentioned [1], which suppresses cell damage caused by transport of zinc, iron, manganese or cadmium via the metal transporter ZIP14.
[11] The inhibitor according to the above-mentioned [1], which does not inhibit the metal transporter ZIP8.
[12] The inhibitor according to the above-mentioned [11], which does not inhibit transport of zinc, iron, manganese or cadmium via the metal transporter ZIP8 and does not suppress cell damage caused by said transport.
[13] A preventive or therapeutic agent for a disease associated with the metal transporter ZIP14, comprising a compound defined in any one of the above [1] to [12] or a salt thereof.
[14] The preventive or therapeutic agent according to the above-mentioned [13], wherein the disease associated with the metal transporter ZIP14 is cancer cachexia accompanied by skeletal muscle atrophy or loss, iron overload disease, liver cancer, or manganese or cadmium toxicity.

[15] A pharmaceutical composition for inhibiting the metal transporter ZIP14, comprising a compound defined in any one of the above [1] to [12] or a salt thereof, and a pharma- ceutically acceptable carrier.
[16] A pharmaceutical composition for preventing or treating a disease associated with the metal transporter ZIP14, comprising a compound defined in any one of the above [1] to [12] or a salt thereof, and a pharma- ceutically acceptable carrier.

[17] 8-(2-(3-methylphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 7);
3-Methyl-1-phenyl-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 9);
1-Phenyl-8-(2-phenylethyl)-3-propyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 10);
3-benzyl-1-phenyl-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 11);
1-(4-methoxyphenyl)-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 14);
8-(2-(4-azidophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 24);
8-(2-(4-biphenylyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 25);
8-(2-(2-naphthyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 26);
8-(2-(2,4-dimethylphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 31);
1-Phenyl-8-(2-(4-sulfamoylphenyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 34);
8-(2-(3-azidophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 36);
8-(2-(2-azidophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 37);
1-Phenyl-8-(2-(5-pyrimidinyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 38); and 1-Phenyl-8-(2-(4-hydroxy-2-methylphenyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 39).
or a salt thereof.

According to the present invention, compound (I) has an inhibitory effect on the metal transporter ZIP14, and is therefore expected to be useful in the prevention or treatment of diseases associated with ZIP14 (e.g., cancer cachexia accompanied by skeletal muscle atrophy or loss, iron overload (including liver damage associated with iron overload), liver cancer, manganese or cadmium poisoning, etc., particularly cancer cachexia accompanied by skeletal muscle atrophy or loss).
Furthermore, compound (I) does not affect metal transport by the metal transporter ZIP8, which is most closely related to the metal transporter ZIP14 in terms of molecular evolution, and therefore serves as a selective inhibitor of the metal transporter ZIP14.

FIG. 1 shows that compound 3 (PPTD) inhibits ZIP14-mediated transport of zinc (A) or iron (B). FIG. 2 shows that compound 3 (PPTD) inhibits ZIP14-mediated transport of manganese (A) or cadmium (B). FIG. 3 shows that compound 3 (PPTD) suppresses cell damage caused by zinc transport via ZIP14. FIG. 4 shows that compound 3 (PPTD) suppresses cell damage caused by zinc transport via ZIP14. FIG. 5 shows that compound 3 (PPTD) suppresses cell injury due to iron transport via ZIP14. FIG. 6 shows that compound 3 (PPTD) suppresses cell damage caused by manganese (A) or cadmium (B) transport via ZIP14. FIG. 7 shows that compound 3 (PPTD) does not inhibit ZIP8-mediated transport of zinc (A) or iron (B). FIG. 8 shows that compound 3 (PPTD) does not inhibit ZIP8-mediated transport of manganese (A) or cadmium (B). FIG. 9 shows that compound 3 (PPTD) does not suppress cell damage caused by zinc transport via ZIP8. FIG. 10 shows that compound 3 (PPTD) does not suppress cell damage caused by iron transport via ZIP8. FIG. 11 shows that compound 3 (PPTD) does not suppress cell damage caused by manganese (A) or cadmium (B) transport via ZIP8. FIG. 12 shows that compound 3 (PPTD) inhibits the proliferation of HepG2 cells, a cell line derived from human hepatic cancer. FIG. 13 shows that compound 3 hydrochloride (PPTD-HCl) inhibits iron transport via ZIP14 equivalently to compound 3 (PPTD). FIG. 14 is a diagram showing that compound 3 (PPTD) irreversibly inhibits iron transport via ZIP14 ( ⁵⁵ Fe radioactivity). FIG. 15 is a graph (relative values) showing that compound 3 (PPTD) irreversibly inhibits iron transport via ZIP14. FIG. 16 shows that compound 3 (PPTD) inhibits ZIP14-mediated production of reactive oxygen species (ROS). FIG. 17 shows that compound 3 (PPTD) inhibits the production of lipid peroxide (LPO) mediated by ZIP14.

The present invention will be described in detail below.
The definition of each substituent used in the present specification is described in detail below. Unless otherwise specified, each substituent has the following definition.

In this specification, "halogen atom" means a fluorine atom, chlorine atom, bromine atom, or iodine atom.

In the present specification, a " _C1-6 alkyl group" refers to a linear or branched saturated hydrocarbon group having 1 to 6 carbon atoms. Examples of the " _C1-6 alkyl group" include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, 1-ethylpropyl, hexyl, isohexyl, 1,1-dimethylbutyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, 2-ethylbutyl and the like, with a _C1-4 alkyl group being preferred.

In the present specification, a "C _1-4 alkyl group" means a linear or branched saturated hydrocarbon group having 1 to 4 carbon atoms. Examples of the "C _1-4 alkyl group" include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, and tert-butyl.

In the present specification, "C _1-6 alkoxy group" refers to a group represented by the formula -OR ¹¹ (wherein R ¹¹ represents a C _1-6 alkyl group). Examples of the "C _1-6 alkoxy group" include methoxy, ethoxy, propoxy, isopropoxy, butyloxy, isobutyloxy, sec-butyloxy, tert-butyloxy, pentyloxy, hexyloxy and the like, with a C _1-4 alkoxy group being preferred.

In the present specification, a " _C1-6 haloalkyl group" refers to a group in which one or more hydrogen atoms in a " _C1-6 alkyl group" have been substituted with a halogen atom. Examples of the " _C1-6 haloalkyl group" include fluoromethyl, 2-fluoroethyl, 3-fluoropropyl, 4-fluorobutyl, trifluoromethyl, difluoromethyl, perfluoroethyl, perfluoropropyl, chloromethyl, 2-chloroethyl, bromomethyl, 2-bromoethyl, iodomethyl, 2-iodoethyl and the like, with trifluoromethyl being preferred.

As used herein, the term "C _1-6 haloalkoxy group" refers to a group in which one or more hydrogen atoms in a "C _1-6 alkoxy group" have been substituted with a halogen atom. Examples of the "C _1-6 haloalkoxy group" include bromomethoxy, 2-bromoethoxy, 3-bromopropoxy, 4-bromobutoxy, iodomethoxy, 2-iodoethoxy, 3-iodopropoxy, 4-iodobutoxy, fluoromethoxy, 2-fluoroethoxy, 3-fluoropropoxy, 4-fluorobutoxy, tribromomethoxy, trichloromethoxy, trifluoromethoxy, difluoromethoxy, perfluoroethoxy, perfluoropropoxy, perfluoroisopropoxy, 1,1,2,2-tetrafluoroethoxy, 2-chloro-1,1,2-trifluoroethoxy and the like.

As used herein, a "mono-C _1-6 alkylamino group" refers to a group represented by the formula -NHR ¹¹ (wherein R ¹¹ represents a C _1-6 alkyl group). Examples of the "mono-C _1-6 alkylamino group" include methylamino, ethylamino, propylamino, isopropylamino, butylamino, isobutylamino, sec-butylamino, tert-butylamino, pentylamino, hexylamino and the like, with a mono-C _1-4 alkylamino group being preferred.

In the present specification, a "di-C _1-6 alkylamino group" refers to a group represented by the formula -NR ¹¹ ₂ (wherein each of the two R ¹¹ independently represents a C _1-6 alkyl group). Examples of the "di-C _1-6 alkylamino group" include dimethylamino, diethylamino, N-ethyl-N-methylamino, dipropylamino, diisopropylamino, dibutylamino, diisobutylamino, disec-butylamino, ditert-butylamino, dipentylamino, dihexylamino and the like, with a di-C _1-4 alkylamino group being preferred.

In the present specification, the term "C _1-6 alkyl-carbonyl group" refers to a group represented by the formula -C(=O)R ¹¹ (wherein R ¹¹ represents a C _1-6 alkyl group). Examples of the "C _{1-6 alkyl-carbonyl group" include acetyl, propanoyl, butanoyl, 2-methylpropanoyl, pentanoyl, 3-methylbutanoyl, 2-methylbutanoyl, 2,2-dimethylpropanoyl, hexanoyl, heptanoyl and the like, with a C 1-4 alkyl} -carbonyl group being preferred.

In the present specification, the term "C _1-6 alkoxy-carbonyl group" refers to a group represented by the formula -C(=O)OR ¹¹ (wherein R ¹¹ represents a C _1-6 alkyl group). Examples of the "C 1-6 alkoxy-carbonyl group" include methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, isobutoxycarbonyl, sec-butoxycarbonyl, tert-butoxycarbonyl, pentyloxycarbonyl, hexyloxycarbonyl and the like, with _{a C 1-4 alkoxy} -carbonyl group being preferred.

In the present specification, a "mono-C _1-6 alkyl-carbamoyl group" refers to a group represented by the formula -C(=O)NHR ¹¹ (wherein R ¹¹ represents a C _1-6 alkyl group). Examples of the "mono-C _1-6 alkyl-carbamoyl group" include methylcarbamoyl, ethylcarbamoyl, propylcarbamoyl, isopropylcarbamoyl, butylcarbamoyl, isobutylcarbamoyl, sec-butylcarbamoyl, tert-butylcarbamoyl, pentylcarbamoyl, hexylcarbamoyl and the like, with a mono-C _1-4 alkyl-carbamoyl group being preferred.

In the present specification, a "di-C _1-6 alkyl-carbamoyl group" refers to a group represented by the formula -C(=O)NR ¹¹ ₂ (wherein each of the two R ¹¹ independently represents a C _1-6 alkyl group). Examples of the "di-C _1-6 alkyl-carbamoyl group" include dimethylcarbamoyl, diethylcarbamoyl, dipropylcarbamoyl, diisopropylcarbamoyl, dibutylcarbamoyl, diisobutylcarbamoyl, disec-butylcarbamoyl, ditert-butylcarbamoyl, dipentylcarbamoyl, dihexylcarbamoyl and the like, with a di-C _1-4 alkyl-carbamoyl group being preferred.

As used herein, a "mono-C _1-6 alkyl-sulfamoyl group" refers to a group represented by the formula -S(=O) ₂ NHR ¹¹ (wherein R ¹¹ represents a C _1-6 alkyl group). Examples of the "mono-C _1-6 alkyl-sulfamoyl group" include methylsulfamoyl, ethylsulfamoyl, propylsulfamoyl, isopropylsulfamoyl, butylsulfamoyl, isobutylsulfamoyl, sec-butylsulfamoyl, tert-butylsulfamoyl, pentylsulfamoyl, hexylsulfamoyl and the like, with a mono-C _1-4 alkyl-sulfamoyl group being preferred.

In the present specification, a "di-C _1-6 alkyl-sulfamoyl group" refers to a group represented by the formula -S(=O) ₂ NR ¹¹ ₂ (wherein each of the two R ¹¹ independently represents a C _1-6 alkyl group). Examples of the "di-C _1-6 alkyl-sulfamoyl group" include dimethylsulfamoyl, diethylsulfamoyl, dipropylsulfamoyl, diisopropylsulfamoyl, dibutylsulfamoyl, diisobutylsulfamoyl, disec-butylsulfamoyl, ditert-butylsulfamoyl, dipentylsulfamoyl, dihexylsulfamoyl and the like, with a di-C _1-4 alkyl-sulfamoyl group being preferred.

In the present specification, a "C _6-10 aryl group" means a hydrocarbon group having aromaticity and having 6 to 10 carbon atoms. Examples of the "C _6-10 aryl group" include phenyl, 1-naphthyl and 2-naphthyl, with phenyl being preferred.

In the present specification, the term " _C7-14 aralkyl group" refers to a " _C1-4 alkyl group" substituted with a " _C6-10 aryl group". Examples of the " _C7-14 aralkyl group" include benzyl, 1-phenylethyl, 2-phenylethyl, 3-phenylpropyl, 4-phenylbutyl, (1-naphthyl)methyl, (2-naphthyl)methyl and the like, with benzyl being preferred.

In the present specification, the term "5- or 6-membered aromatic heterocyclic group" refers to a 5- or 6-membered aromatic group containing at least one heteroatom selected from a nitrogen atom, an oxygen atom, and a sulfur atom. Examples of the "5- or 6-membered aromatic heterocyclic group" include thienyl, furyl, pyrrolyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, triazolyl, tetrazolyl, triazinyl, etc.

In the present specification, the term "3- to 8-membered non-aromatic heterocyclic group" refers to a 3- to 8-membered non-aromatic group containing at least one heteroatom selected from a nitrogen atom, an oxygen atom, and a sulfur atom. Examples of "3- to 8-membered non-aromatic heterocyclic group" include aziridinyl, oxiranyl, thiiranyl, azetidinyl, oxetanyl, thietanyl, tetrahydrothienyl, tetrahydrofuranyl, pyrrolinyl, pyrrolidinyl, imidazolinyl, imidazolidinyl, oxazolinyl, oxazolidinyl, pyrazolinyl, pyrazolidinyl, thiazolinyl, thiazolidinyl, tetrahydroisothiazolyl, tetrahydrooxazolyl, tetrahydroisothiazolyl ... Examples include soxazolyl, piperidinyl, piperazinyl, tetrahydropyridinyl, dihydropyridinyl, dihydrothiopyranyl, tetrahydropyrimidinyl, tetrahydropyridazinyl, dihydropyranyl, tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, thiomorpholinyl, azepanyl, diazepanyl, azepinyl, oxepanyl, azocanyl, and diazocanyl, and 5- or 6-membered non-aromatic heterocyclic groups are preferred.

In the present specification, the term "C _3-8 cycloalkyl group" means a cyclic saturated hydrocarbon group having 3 to 8 carbon atoms. Examples of the "C _3-8 cycloalkyl group" include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl, and a C _5-6 cycloalkyl group is preferable.

In the present specification, a "C _3-8 cycloalkenyl group" means a cyclic unsaturated hydrocarbon group having 3 to 8 carbon atoms. Examples of the "C _3-8 cycloalkenyl group" include cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl and the like, with a C _5-6 cycloalkenyl group being preferred.

In this specification, the term "C _1-6 alkylene group" means a linear or branched divalent saturated hydrocarbon group having 1 to 6 carbon atoms. Examples of the "C _1-6 alkylene group" include, for example, -CH ₂ -, -(CH ₂ ) ₂ -, -(CH ₂ ) ₃ -, -(CH ₂ ) ₄ -, -(CH ₂ ) ₅ -, -(CH ₂ ) ₆ -, -CH(CH ₃ )-, -C(CH ₃ ) ₂ -, -CH(C ₂ H ₅ )-, -CH(C ₃ H ₇ )-, -CH(CH(CH ₃ ) ₂ )-, -(CH(CH ₃ )) ₂ -, -CH ₂ -CH(CH ₃ )-, -CH(CH ₃ )-CH ₂ -, -CH ₂ -CH ₂ -C(CH ₃ ) ₂ -, -C(CH ₃ ) ₂ -CH ₂ -CH ₂ -, -CH ₂ Examples include -CH2 _{- CH2} -C( _CH3 ) _2- and -C( _CH3 ) ₂ -CH2- _CH2 - _CH2 - _CH2- , with a _C1-3 alkylene group being preferred.

The metal transporter ZIP14 inhibitor of the present invention includes compound (I).
In formula (I), R ¹ represents a C _6-10 aryl group, a C _7-14 aralkyl group, a 5- or 6-membered aromatic heterocyclic group, a 3- to 8-membered non-aromatic heterocyclic group, a C _3-8 cycloalkyl group, a C _3-8 cycloalkenyl group, or a C _1-6 alkyl group, wherein the C _6-10 aryl group, C _7-14 aralkyl group, a 5- or 6-membered aromatic heterocyclic group, a 3- to 8-membered non-aromatic heterocyclic group, a C _3-8 cycloalkyl group, or a C _3-8 cycloalkenyl group is optionally substituted by a substituent selected from substituent group Z.

Substituent group Z consists of a halogen atom, a cyano group, a nitro group, a C _1-6 alkyl group, a C _1-6 haloalkyl group, a hydroxy group, a C _1-6 alkoxy group, a C _1-6 haloalkoxy group, an amino group, a mono-C _1-6 alkylamino group, a di-C _1-6 alkylamino group, a carboxy group, a C _1-6 alkyl-carbonyl group, a C _1-6 alkoxy-carbonyl group, an azido group, a carbamoyl group, a mono-C _1-6 alkyl-carbamoyl group, a di-C _1-6 alkyl-carbamoyl group, a sulfamoyl group, a mono-C _{1-6 alkyl-sulfamoyl group, a di-C 1-6 alkyl} -sulfamoyl group, a sulfo group, a methylenedioxy group (-OCH ₂ O-), and a C _6-10 aryl group.

In one embodiment, the substituent group Z preferably consists of a halogen atom, a cyano group, a nitro group, a C _1-2 alkyl group, a C _1-2 haloalkyl group, a hydroxy group, a C _1-2 alkoxy group, a C _1-2 haloalkoxy group, an amino group, a mono-C _1-2 alkylamino group, a di-C _1-2 alkylamino group, a carboxy group, a C _1-2 alkyl-carbonyl group, a C _1-2 alkoxy-carbonyl group, an azido group, a carbamoyl group, a mono-C _1-2 alkyl-carbamoyl group, a di-C _1-2 alkyl-carbamoyl group, a sulfamoyl group, a mono-C _1-2 alkyl-sulfamoyl group, a di-C _1-2 alkyl-sulfamoyl group, a sulfo group, a methylenedioxy group (-OCH ₂ O-), and a phenyl group.

In one embodiment, R ¹ is preferably a C _6-10 aryl group (preferably a phenyl group), a C _7-14 aralkyl group (preferably a benzyl group), a 5- or 6-membered aromatic heterocyclic group, a 3- to 8-membered non-aromatic heterocyclic group (preferably a 5- or 6-membered non-aromatic heterocyclic group), a C _3-8 cycloalkyl group (preferably a C _5-6 cycloalkyl group), or a C _3-8 cycloalkenyl group (preferably a C _5-6 cycloalkenyl group), which may be substituted by a substituent selected from substituent group Z.

In another embodiment, R ¹ is preferably a C _6-10 aryl group (preferably a phenyl group) optionally substituted by a substituent selected from the substituent group Z, or a C _1-6 alkyl group (preferably a methyl group).

In another embodiment, R ¹ is more preferably a phenyl group, a benzyl group, a 5- or 6-membered aromatic heterocyclic group, a 5- or 6-membered non-aromatic heterocyclic group, a C _5-6 cycloalkyl group, or a C _5-6 cycloalkenyl group, which may be substituted with a substituent selected from substituent group Z. ^{R 1} is further preferably a phenyl group, a benzyl group, or a 6-membered aromatic heterocyclic group, which may be substituted with a substituent selected from substituent group Z.

In another embodiment, R ¹ is more preferably a C _6-10 aryl group (preferably a phenyl group) optionally substituted with a substituent selected from the substituent group Z.

R ¹ is more preferably a phenyl group which may be substituted with a substituent selected from the substituent group Z.

The substituent group Z for R ¹ is preferably a halogen atom and a C _1-6 alkoxy group (preferably a C _1-2 alkoxy group), more preferably a halogen atom, particularly preferably a fluorine atom.

R ¹ is even more preferably a phenyl group which may be substituted with a halogen atom (preferably a fluorine atom), and particularly preferably an unsubstituted phenyl group.

In formula (I), R ² represents a hydrogen atom or a C _1-6 alkyl group.
^R2 is preferably a hydrogen atom.

In formula (I), R ³ represents a hydrogen atom, a C _1-6 alkyl group, a C _6-10 aryl group, a C _7-14 aralkyl group, a 5- or 6-membered aromatic heterocyclic group, a 3- to 8-membered non-aromatic heterocyclic group, a C _3-8 cycloalkyl group, or a C _3-8 cycloalkenyl group, wherein the C _6-10 aryl group, C _7-14 aralkyl group, a 5- or 6-membered aromatic heterocyclic group, a 3- to 8-membered non-aromatic heterocyclic group, a C _3-8 cycloalkyl group, or a C _3-8 cycloalkenyl group is optionally substituted by a substituent selected from substituent group Z.
R ³ is preferably a hydrogen atom, a C _1-6 alkyl group (preferably a C _1-2 alkyl group) or a C _7-14 aralkyl group (preferably a benzyl group) (the C _7-14 aralkyl group may be substituted with a substituent selected from substituent group Z), more preferably a hydrogen atom or a C _1-6 alkyl group (preferably a C _1-2 alkyl group), even more preferably a hydrogen atom or a C _1-2 alkyl group (preferably a methyl group), still more preferably a hydrogen atom or a methyl group, and particularly preferably a hydrogen atom.

In formula (I), R ⁴ is selected from a halogen atom, a cyano group, a nitro group, a C _1-6 alkyl group, a C _1-6 haloalkyl group, a hydroxy group, a C _1-6 alkoxy group, a C _1-6 haloalkoxy group, an amino group, a mono-C _1-6 alkylamino group, a di-C _1-6 alkylamino group, a carboxy group, a C _1-6 alkyl-carbonyl group, and a C _1-6 alkoxy-carbonyl group, and n represents an integer from 0 to 2.
n is preferably 0.

In formula (I), R ⁵ represents a hydrogen atom, a C _1-6 alkyl group, or a group represented by the formula: -L-Cy.
^R5 is preferably a hydrogen atom or a group represented by the formula: -L-Cy.
^R5 is preferably a group represented by the formula: -L-Cy.

L represents a bond, a C _1-6 alkylene group, or a group represented by the formula: *-(CH ₂ ) _m1 -X-(CH ₂ ) _m2 -**.
Here, X represents O, S, NR ⁶ , CO, NR ⁶ CO, CONR ⁶ , COO, or OCO, R ⁶ represents a hydrogen atom or a C _1-6 alkyl group, m1 represents an integer of 0 to 3, m2 represents an integer of 0 to 3, * represents a bonding site to the nitrogen atom of the spiro ring, and ** represents a bonding site to Cy. R ⁶ is preferably a hydrogen atom or a methyl group.
Examples of the group represented by *-(CH ₂ ) _m1 -X-(CH ₂ ) _m2 -** include
*-(CH ₂ ) _m1 -O-**, *-(CH ₂ ) _m1 -O-(CH ₂ ) _m2 -**;
*-(CH ₂ ) _m1 -S-**, *-(CH ₂ ) _m1 -S-(CH ₂ ) _m2 -**;
*-(CH ₂ ) _m1 -NH-**, *-(CH ₂ ) _m1 -NH-(CH ₂ ) _m2 -**, *-(CH ₂ ) _m1 -N(CH ₃ )-**, *-(CH ₂ ) _m1 -N(CH ₃ )-(CH ₂ ) _m2 -**;
*-(CH ₂ ) _m1 -CO-**, *-(CH ₂ ) _m1 -CO-(CH ₂ ) _m2 -**; *-(CH ₂ ) _m1 -NHCO-**, *-(CH ₂ ) _m1 -NHCO-(CH ₂ ) _m2 -**, *-(CH ₂ ) _m1 - N(CH ₃ )CO-**, *-(CH ₂ ) _m1 -N(CH ₃ )CO-(CH ₂ ) _m2 -**;
*-(CH ₂ ) _m1 -CONH-**, *-(CH ₂ ) _m1 -CONH-(CH ₂ ) _m2 -**, *-(CH ₂ ) _m1 -CON(CH ₃ )-**, *-(CH ₂ ) _m1 -CON(CH ₃ )-(CH ₂ ) _m2 -* *；
*-(CH ₂ ) _m1 -COO-**, *-(CH ₂ ) _m1 -COO-(CH ₂ ) _m2 -**;
*-(CH ₂ ) _m1 -OCO-**, *-(CH ₂ ) _m1 -OCO-(CH ₂ ) _m2 -**;
etc.
L is preferably a bond or a C _1-6 alkylene group, more preferably a C _1-6 alkylene group, even more preferably a C _1-3 alkylene group, still more preferably ethylene (-CH ₂ -CH ₂ -) or trimethylene (-CH ₂ -CH ₂ -CH ₂ -), and particularly preferably ethylene (-CH ₂ -CH ₂ -).

Cy represents a C _6-10 aryl group (preferably a phenyl group, a naphthyl group), a 5- or 6-membered aromatic heterocyclic group (preferably a 5- or 6-membered nitrogen-containing aromatic heterocyclic group), a 3- to 8-membered non-aromatic heterocyclic group (preferably a 5- or 6-membered non-aromatic heterocyclic group), a C _3-8 cycloalkyl group (preferably a C _5-6 cycloalkyl group), or a C _3-8 cycloalkenyl group (preferably a C _5-6 cycloalkenyl group), wherein the C _6-10 aryl group, the 5- or 6-membered aromatic heterocyclic group, the 3- to 8-membered non-aromatic heterocyclic group, the C _3-8 cycloalkyl group, or the C _3-8 cycloalkenyl group is optionally substituted by a substituent selected from substituent group Z.

In one embodiment, Cy is preferably a C _6-10 aryl group (preferably a phenyl group, a naphthyl group), a 5- or 6-membered aromatic heterocyclic group (preferably a 5- or 6-membered nitrogen-containing aromatic heterocyclic group), or a C _3-8 cycloalkyl group (preferably a C _5-6 cycloalkyl group), which may be substituted by a substituent selected from substituent group Z.

In another embodiment, Cy is preferably a phenyl group, a 5- or 6-membered aromatic heterocyclic group (preferably a 5- or 6-membered nitrogen-containing aromatic heterocyclic group), a 5- or 6-membered non-aromatic heterocyclic group, a C _5-6 cycloalkyl group, or a C _5-6 cycloalkenyl group, which may be substituted by a substituent selected from substituent group Z.

Cy is more preferably a phenyl group, a 5- or 6-membered nitrogen-containing aromatic heterocyclic group (preferably a pyridyl group, a pyrimidinyl group), or a C _5-6 cycloalkyl group, which may be substituted with a substituent selected from substituent group Z.
Cy is more preferably a phenyl group, a 6-membered nitrogen-containing aromatic heterocyclic group (preferably a pyridyl group or a pyrimidinyl group), or a cyclohexyl group, which may be substituted with a substituent selected from the substituent group Z.
Cy is even more preferably a phenyl group which may be substituted with a substituent selected from the substituent group Z.

The substituent group Z for Cy preferably consists of a halogen atom (preferably a fluorine atom, a chlorine atom), a nitro group, a C _1-6 alkyl group (preferably a C _1-2 alkyl group), a hydroxy group, a C _1-6 alkoxy group (preferably a C _1-2 alkoxy group), an amino group, a carboxy group, an azide group, a carbamoyl group, a sulfamoyl group, a sulfo group, a methylenedioxy group (-OCH ₂ O-), and a C _6-10 aryl group (preferably a phenyl group).
The substituent group Z for Cy more preferably consists of a halogen atom (preferably a fluorine atom), a C _1-6 alkyl group (preferably a C _1-2 alkyl group), a hydroxy group, a C _1-6 alkoxy group (preferably a C _1-2 alkoxy group), an amino group, an azide group, a sulfamoyl group, and a C _6-10 aryl group (preferably a phenyl group).
The substituent group Z for Cy more preferably consists of a halogen atom (preferably a fluorine atom), a C _1-2 alkyl group, a hydroxy group, a C _1-2 alkoxy group, an amino group, an azide group, and a sulfamoyl group.
The substituent group Z for Cy even more preferably consists of a fluorine atom, a methyl group, a hydroxy group and an azide group.
The substituent group Z for Cy still more preferably consists of a methyl group, a hydroxy group and an azide group.

Preferred examples of the compound represented by formula (I) include the following compounds:

[Wherein,
R ^3a represents a hydrogen atom or a C _1-2 alkyl group;
R ⁷ and R ⁸ each independently represent a group selected from the substituent group Z;
m3 represents 1 to 6;
p represents 0 to 5;
q represents 0 to 5.
A compound having a benzene ring represented by the formula:

[In the formula, r represents 1 or 2;
The other symbols are as defined above.]
or a compound in which the saturated hydrocarbon ring is unsaturated;

[Wherein,
X ¹ represents O, S or NH;
X ² , X ³ , X ⁴ and X ⁵ each independently represent N or CH;
The other symbols are as defined above.]
or a compound having a five-membered aromatic heterocycle represented by the following formula:

[Each symbol in the formula has the same meaning as defined above.]
or a compound having a 6-membered aromatic heterocycle represented by the following formula:

[Wherein,
^X6 represents CH or N;
X ⁷ , X ⁸ , X ⁹ , X ¹⁰ and X ¹¹ each independently represent O, S or NH;
The other symbols are as defined above.]
or a compound having a 6-membered saturated heterocyclic ring represented by the following formula:

[Wherein,
X ¹² , X ¹³ , X ¹⁴ and X ¹⁵ each independently represent O or S;
The other symbols are as defined above.]
or a compound having a 5-membered saturated heterocyclic ring represented by the following formula:
etc.

In the above formula, R ^3a is preferably a hydrogen atom or a methyl group, and particularly preferably a hydrogen atom.

R ⁷ is preferably each independently a halogen atom, and particularly preferably a fluorine atom.

R ⁸ is preferably, each independently, a halogen atom (preferably a fluorine atom, a chlorine atom), a nitro group, a C _1-6 alkyl group (preferably a C _1-2 alkyl group), a hydroxy group, a C _1-6 alkoxy group (preferably a C _1-2 alkoxy group), an amino group, a carboxy group, an azido group, a carbamoyl group, a sulfamoyl group, a sulfo group, a methylenedioxy group (-OCH ₂ O-), or a C _6-10 aryl group (preferably a phenyl group).
R ⁸ is more preferably each independently a halogen atom (preferably a fluorine atom), a C _1-6 alkyl group (preferably a C _1-2 alkyl group), a hydroxy group, a C _1-6 alkoxy group (preferably a C _1-2 alkoxy group), an amino group, an azido group, a sulfamoyl group, or a C _6-10 aryl group (preferably a phenyl group).
R ⁸ is more preferably each independently a halogen atom (preferably a fluorine atom), a C _1-2 alkyl group, a hydroxy group, a C _1-2 alkoxy group, an amino group, an azide group, or a sulfamoyl group, even more preferably each independently a fluorine atom, a methyl group, a hydroxy group, or an azide group, and even more preferably each independently a methyl group, a hydroxy group, or an azide group.

In the above formula, m3 is preferably 1 to 3, more preferably 2 or 3, and particularly preferably 2.
In the above formula, p is preferably 0 to 2, more preferably 0 or 1, and particularly preferably 0.
In the above formula, q is preferably 0 to 2.

Preferable examples of the compound represented by formula (I) include the following compounds I-A to I-C.

Compound I-A
R ¹ is a C _6-10 aryl group (preferably a phenyl group), a C _7-14 aralkyl group (preferably a benzyl group), a 5- or 6-membered aromatic heterocyclic group, a 3- to 8-membered non-aromatic heterocyclic group (preferably a 5- or 6-membered non-aromatic heterocyclic group), a C _3-8 cycloalkyl group (preferably a C _5-6 cycloalkyl group), or a C _3-8 cycloalkenyl group (preferably a C _5-6 cycloalkenyl group), which may be substituted by a substituent selected from substituent group Z;
^R2 is a hydrogen atom;
R ³ is a hydrogen atom or a C _1-2 alkyl group (preferably a methyl group) (preferably a hydrogen atom or a methyl group, particularly preferably a hydrogen atom);
n is 0;
^R5 is a group of the formula: -L-Cy;
L is a C _1-6 alkylene group (preferably a C _1-3 alkylene group), and
Cy is a C _6-10 aryl group (preferably a phenyl group, a naphthyl group), a 5- or 6-membered aromatic heterocyclic group (preferably a 5- or 6-membered nitrogen-containing aromatic heterocyclic group), a 3- to 8-membered non-aromatic heterocyclic group (preferably a 5- or 6-membered non-aromatic heterocyclic group), a C _3-8 cycloalkyl group (preferably a C _5-6 cycloalkyl group) or a C _3-8 cycloalkenyl group (preferably a C _5-6 cycloalkenyl group), which are optionally substituted by a substituent selected from substituent group Z;
Compound (I).

Compound I-B
R ¹ is a C _6-10 aryl group (preferably a phenyl group) optionally substituted by a substituent selected from the substituent group Z;
^R2 is a hydrogen atom;
R ³ is a hydrogen atom or a C _1-2 alkyl group (preferably a methyl group) (preferably a hydrogen atom or a methyl group, particularly preferably a hydrogen atom);
n is 0;
^R5 is a group of the formula: -L-Cy;
L is a C _1-6 alkylene group (preferably a C _1-3 alkylene group), and
Cy is a phenyl group, a 5- or 6-membered aromatic heterocyclic group (preferably a 5- or 6-membered nitrogen-containing aromatic heterocyclic group), a 5- or 6-membered non-aromatic heterocyclic group, a C _5-6 cycloalkyl group, or a C _5-6 cycloalkenyl group, which may be substituted by a substituent selected from substituent group Z;
Compound (I).

Compound I-C
R ¹ is a phenyl group optionally substituted with a substituent selected from the substituent group Z (preferably a halogen atom, more preferably a fluorine atom);
^R2 is a hydrogen atom;
R ³ is a hydrogen atom or a methyl group (preferably a hydrogen atom);
n is 0;
^R5 is a group of the formula: -L-Cy;
L is a C _1-3 alkylene group; and
Cy is a phenyl group, a 5- or 6-membered nitrogen-containing aromatic heterocyclic group (preferably a pyridyl group, a pyrimidinyl group), or a C _5-6 cycloalkyl group, which may be substituted by a substituent selected from the substituent group Z;
Compound (I).

Specific examples of compound (I) include the following compounds.
1-Phenyl-1,3,8-triazaspiro[4.5]decan-4-one (Compound 1);
8-Benzyl-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 2);
1-phenyl-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 3) (also known as PPTD);
1-Phenyl-8-(3-phenylpropyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 4);
8-(2-(4-methoxyphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 5);
8-(2-(4-fluorophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 6);
8-(2-(3-methylphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 7);
8-(2-cyclohexylethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 8);
3-Methyl-1-phenyl-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 9);
1-Phenyl-8-(2-phenylethyl)-3-propyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 10);
3-benzyl-1-phenyl-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 11);
8-benzyl-1-(4-methoxyphenyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 12);
1-(4-Methoxyphenyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 13);
1-(4-methoxyphenyl)-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 14);
8-benzyl-1-(4-fluorophenyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 15);
1-(4-fluorophenyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 16);
1-(4-fluorophenyl)-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 17);
8-Benzyl-1-methyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 18);
1-Methyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 19);
1-Methyl-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 20);
8-(2-(4-methylphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 21);
8-(2-(2-methylphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 22);
1-Phenyl-8-(2-phenylpropyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 23);
8-(2-(4-azidophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 24);
8-(2-(4-biphenylyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 25);
8-(2-(2-naphthyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 26);
1-Phenyl-8-(2-(4-pyridyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 27);
1-Phenyl-8-(2-(3-pyridyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 28);
1-Phenyl-8-(2-(2-pyridyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 29);
8-(3,3-diphenylpropyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 30);
8-(2-(2,4-dimethylphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 31);
8-(2-(3,5-dimethylphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 32);
8-(2-(4-hydroxyphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 33);
1-Phenyl-8-(2-(4-sulfamoylphenyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 34);
8-(2-(4-aminophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 35);
8-(2-(3-azidophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 36);
8-(2-(2-azidophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 37);
1-Phenyl-8-(2-(5-pyrimidinyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 38); and 1-Phenyl-8-(2-(4-hydroxy-2-methylphenyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 39).
and their salts.

Other examples include compounds A2 to A30 shown in Table 2 below.

Among others, it is particularly noteworthy for its excellent ZIP14 inhibitory activity.
1-phenyl-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 3) (also known as PPTD);
1-Phenyl-8-(3-phenylpropyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 4);
8-(2-(4-methoxyphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 5);
8-(2-(4-fluorophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 6);
8-(2-(3-methylphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 7);
8-(2-cyclohexylethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 8);
3-Methyl-1-phenyl-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 9);
1-(4-fluorophenyl)-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 17);
8-(2-(4-methylphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 21);
8-(2-(2-methylphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 22);
1-Phenyl-8-(2-phenylpropyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 23);
8-(2-(4-azidophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 24);
1-Phenyl-8-(2-(4-pyridyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 27);
1-Phenyl-8-(2-(3-pyridyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 28);
1-Phenyl-8-(2-(2-pyridyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 29);
8-(2-(2,4-dimethylphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 31);
8-(2-(3,5-dimethylphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 32);
8-(2-(4-hydroxyphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 33);
1-Phenyl-8-(2-(4-sulfamoylphenyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 34);
8-(2-(4-aminophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 35);
8-(2-(3-azidophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 36);
8-(2-(2-azidophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 37);
1-Phenyl-8-(2-(5-pyrimidinyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 38); and 1-Phenyl-8-(2-(4-hydroxy-2-methylphenyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 39).
and salts thereof are preferred.

Furthermore, it exhibits excellent ZIP14 inhibitory activity,
1-phenyl-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 3) (also known as PPTD);
8-(2-(4-fluorophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 6);
8-(2-(2-methylphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 22);
8-(2-(2,4-dimethylphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 31);
8-(2-(4-hydroxyphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 33);
8-(2-(3-azidophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 36); and 8-(2-(2-azidophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 37); and 1-phenyl-8-(2-(4-hydroxy-2-methylphenyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 39).
and salts thereof are more preferred.

In one embodiment,
1-phenyl-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 3) (also known as PPTD);
8-(2-(2-methylphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 22);
8-(2-(2,4-dimethylphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 31);
8-(2-(4-hydroxyphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 33);
8-(2-(3-azidophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 36); and 8-(2-(2-azidophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 37); and 1-phenyl-8-(2-(4-hydroxy-2-methylphenyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 39).
and salts thereof are even more preferred.

In one embodiment,
1-phenyl-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 3) (also known as PPTD);
8-(2-(2-azidophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 37); and 1-phenyl-8-(2-(4-hydroxy-2-methylphenyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 39).
and salts thereof are particularly preferred.

In another embodiment, 1-phenyl-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 3) (also known as PPTD) and its salts (particularly the hydrochloride salt) are preferred.

Also,
8-(2-(3-methylphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 7);
3-Methyl-1-phenyl-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 9);
1-Phenyl-8-(2-phenylethyl)-3-propyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 10);
3-benzyl-1-phenyl-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 11);
1-(4-methoxyphenyl)-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 14);
8-(2-(4-azidophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 24);
8-(2-(4-biphenylyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 25);
8-(2-(2-naphthyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 26);
8-(2-(2,4-dimethylphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 31);
1-Phenyl-8-(2-(4-sulfamoylphenyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 34);
8-(2-(3-azidophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 36);
8-(2-(2-azidophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 37);
1-Phenyl-8-(2-(5-pyrimidinyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 38); and 1-Phenyl-8-(2-(4-hydroxy-2-methylphenyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 39).
and the salts thereof are novel compounds.

When compound (I) is a salt, such salts include, for example, metal salts, ammonium salts, salts with organic bases, salts with inorganic acids, salts with organic acids, salts with basic or acidic amino acids, etc. Suitable examples of metal salts include, for example, alkali metal salts such as sodium salts and potassium salts; alkaline earth metal salts such as calcium salts, magnesium salts and barium salts; and aluminum salts. Suitable examples of salts with organic bases include, for example, salts with trimethylamine, triethylamine, pyridine, picoline, 2,6-lutidine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, N,N'-dibenzylethylenediamine, etc. Suitable examples of salts with inorganic acids include, for example, salts with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, etc. Suitable examples of salts with organic acids include salts with formic acid, acetic acid, trifluoroacetic acid, phthalic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, etc. Suitable examples of salts with basic amino acids include salts with arginine, lysine, ornithine, etc., and suitable examples of salts with acidic amino acids include salts with aspartic acid, glutamic acid, etc.
Among these, pharma- ceutically acceptable salts are preferred, including inorganic salts such as alkali metal salts (e.g., sodium salt, potassium salt, etc.) and alkaline earth metal salts (e.g., calcium salt, magnesium salt, barium salt, etc.), ammonium salts, salts with inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, and salts with organic acids such as acetic acid, phthalic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, methanesulfonic acid, p-toluenesulfonic acid, and the like.
From the viewpoint of solubility in water, it is preferably in the form of a salt, and more preferably in the form of a hydrochloride. Improved solubility is expected to lead to improved medicinal efficacy.

When compound (I) has isomers such as tautomers, optical isomers, stereoisomers, positional isomers, and rotational isomers, any one of the isomers and a mixture thereof are included in the compound of the present invention. Furthermore, when compound (I) has optical isomers, optical isomers resolved from the racemate are also included in compound (I).
In addition, compound (I) has a 1,3,8-triazaspiro[4.5]decane skeleton, and isomers based on the spiro atom in the skeleton are also included in compound (I).
Compound (I) may be in the form of a crystal, and both a single crystal form and a mixture of crystal forms are included in Compound (I).
Compound (I) may be a solvate (for example, a hydrate) or a non-solvate, and both are encompassed in compound (I).
Compounds labeled or substituted with isotopes (e.g., ^2H , ^3H , ^11C , ^14C , ^18F , ^35S , ^125I , etc.) are also encompassed by compound (I).

Compound (I) can be produced, for example, by the methods described in Patent Documents 1 to 8 or methods equivalent thereto.
For example, compound (Ic), which is compound (I) in which R ² is a hydrogen atom and n is 0, can be produced by the method shown in Scheme 1.

[In the formula, ^R3a represents ^R3 other than a hydrogen atom, ^R5a represents ^R5 other than a hydrogen atom, ^Hal1 and ^Hal2 each represent a bromine atom or an iodine atom, and the other symbols are as defined above.]
Compound (Ic) can be produced by reacting compound (Ia), which is compound (I) in which R ² and R ⁵ are hydrogen atoms and n is 0, with a compound represented by R ^5a -Hal ¹ in a solvent in the presence of a base.
Compound (Ia) can be produced by a method known per se or a method analogous thereto, or by the method shown in Scheme 2 described below.
The compound represented by R ^5a -Hal ¹ is a commercially available product, or can be produced by a method known per se or a method analogous thereto.
The solvent may be a nitrile solvent such as acetonitrile.
Examples of the base include calcium carbonate, sodium carbonate, and sodium hydrogen carbonate.
When Hal ¹ is a bromine atom, the reaction may be carried out in the presence of potassium iodide.

Alternatively, compound (Ic) can also be produced by reacting compound (Ib), which is compound (I) in which R ² and R ³ are hydrogen atoms and n is 0, with a compound represented by R ^3a -Hal ² in a solvent in the presence of a base.
Compound (Ib) can be produced by a method known per se or a method analogous thereto, or by the method shown in Scheme 2 described below.
The compound represented by R ^3a -Hal ² is a commercially available product, or can be produced by a method known per se or a method analogous thereto.
The solvent may be an ether solvent such as tetrahydrofuran.
The base includes sodium hydride and the like.

[In the formula, P represents an amino-protecting group, and the other symbols are as defined above.]
Examples of the amino-protecting group include a benzyl (Bn) group, a tert-butoxycarbonyl (Boc) group, a benzyloxycarbonyl (Cbz) group, a 9-fluorenylmethyloxycarbonyl (Fmoc) group, etc. When P is a benzyl group, it corresponds to ^R5 .

Compound (2) can be produced by reacting compound (1) with a compound represented by R ¹ -NH ₂ and a cyanating agent in a solvent.
Compound (1) may be a commercially available product or may be produced by a method known per se or a method analogous thereto.
The compound represented by R ¹ -NH ₂ may be a commercially available product or may be produced by a method known per se or a method analogous thereto.
The cyanating agent may, for example, be trimethylsilyl cyanide (Me ₃ SiCN).
The solvent may be acetic acid or the like.

Compound (3) can be produced by reacting compound (2) with an acid.
The acid includes sulfuric acid and the like.
The reaction is usually carried out without a solvent, in which case the above acid is used in an amount equivalent to that of the solvent.

Compound (4) can be produced by subjecting compound (3) to a cyclization reaction.
The cyclization reaction can be carried out by reacting compound (3) with N,N-dimethylformamide dimethyl acetal (DMF-DMA) in a solvent under a nitrogen atmosphere.
The solvent may be an alcohol solvent such as methanol.

Compound (5) can be produced by subjecting compound (4) to a reduction reaction.
The reduction reaction can be carried out by reacting compound (4) with a reducing agent in a solvent under a nitrogen atmosphere.
The reducing agent includes sodium borohydride, lithium aluminum hydride, sodium triacetoxyborohydride, sodium cyanoborohydride, and the like.
The solvent may be an alcohol solvent such as methanol.

Compound (6) can be produced by reacting compound (5) with a compound represented by R ^3a -Hal ² in a solvent in the presence of a base.
The reaction can be carried out in the same manner as described in the production of compound (Ic) from compound (Ib) in Scheme 1.

Compound (Ia) can be produced by subjecting compound (6) to a deprotection reaction.
For example, when the amino-protecting group P is a benzyl (Bn) group, the deprotection reaction can be carried out by subjecting compound (6) to catalytic hydrogenation using palladium on carbon (Pd/C) in a solvent.
The solvent may be an alcohol solvent such as methanol.
The catalytic hydrogenation reaction may be carried out in the presence of an acid such as acetic acid or hydrochloric acid.

Compound (Id), which is compound (I) in which R ² , R ³ and R ⁵ are hydrogen atoms and n is 0, can be produced by subjecting compound (5) to a deprotection reaction.
The reaction can be carried out in the same manner as described in the production of compound (Ia) from compound (6).

Compound (Ib) can be produced by reacting compound (Id) with a compound represented by R ^5a -Hal ¹ in a solvent in the presence of a base.
The reaction can be carried out in the same manner as described in the production of compound (Ic) from compound (Ia) in Scheme 1.

The raw materials and reagents used in each step of the manufacturing method described below, as well as the resulting compounds, may each be a salt. Examples of such salts include the same salts as those of compound (I).

When the compound obtained in each step is a free compound, it can be converted into the desired salt by a method known per se. Conversely, when the compound obtained in each step is a salt, it can be converted into a free form or another type of desired salt by a method known per se.

The compounds obtained in each step can be used in the next reaction either as the reaction solution or as a crude product, or can be isolated and/or purified from the reaction mixture by a separation method such as concentration, crystallization, recrystallization, distillation, solvent extraction, fractional distillation, or chromatography in accordance with conventional methods.

Compound (I) has excellent ZIP14 inhibitory activity. Moreover, the ZIP14 inhibitory activity is irreversible. Therefore, it is expected to prevent or treat diseases related to ZIP14 (e.g., cancer cachexia accompanied by skeletal muscle atrophy or loss, iron overload (including liver damage associated with iron overload), liver cancer, manganese or cadmium poisoning, etc., particularly cancer cachexia accompanied by skeletal muscle atrophy or loss).
Compound (I) also has excellent ZIP14 inhibitory activity, and is therefore expected to suppress cell damage caused by metals such as zinc, iron, manganese, and cadmium that are transported via ZIP14.
Compound (I) further inhibits the production of reactive oxygen species (ROS) and the production of lipid peroxide (LPO) via ZIP14, suggesting that the production of ROS and LPO is involved in cell damage caused by metals such as zinc, iron, manganese, and cadmium transported via ZIP14.
Compound (I) inhibits the metal transport activity of ZIP14, but does not affect the metal transport of ZIP8, which is the closest relative in terms of molecular evolution. At present, the mechanism of metal ion transport via metal transporters has not been elucidated, so compound (I) is expected to become a new research reagent for analyzing the transport mechanism and molecular structure of metal transporters. We believe that the results will greatly contribute to the understanding of ZIP14, a disease-related molecule.

Compound (I) can be safely administered orally or parenterally to mammals (e.g., humans, monkeys, cows, horses, pigs, mice, rats, hamsters, rabbits, cats, dogs, sheep, goats, etc.) as a medicine as it is, or as a pharmaceutical composition mixed with a pharma- ceutical acceptable carrier, etc. "Parenteral" includes administration intravenously, intramuscularly, subcutaneously, intraviscerally, intranasally, intradermally, by eye drop, intracerebral, intrarectally, intravaginally, intraperitoneally, inside a tumor, proximal to a tumor, etc., and administration directly to a lesion.

The dosage of compound (I) varies depending on the administration route, symptoms, etc., but for example, when orally administered to a patient with cancer cachexia (adult, body weight 40-80 kg, e.g. 60 kg), the dosage is, for example, 0.001-1000 mg/kg body weight per day, preferably 0.01-100 mg/kg body weight per day, and more preferably 0.1-10 mg/kg body weight per day. This amount can be administered in one to three divided doses per day.

The pharmaceutical preparation containing compound (I) can be prepared according to a method known per se (e.g., a method described in the Japanese Pharmacopoeia, etc.) as a method for producing pharmaceutical preparations, either alone or as a pharmaceutical composition in which compound (I) is mixed with a pharmaceutical carrier. Pharmaceutical preparations containing compound (I) can be prepared in the form of, for example, tablets (including sugar-coated tablets, film-coated tablets, sublingual tablets, orally disintegrating tablets, buccal tablets, etc.), pills, powders, granules, capsules (including soft capsules and microcapsules), troches, syrups, liquids, emulsions, suspensions, controlled-release preparations (e.g., immediate-release preparations, sustained-release preparations, sustained-release microcapsules), aerosols, films (e.g., orally disintegrating films ... tablets, orally disintegrating tablets, orally disintegrating tablets, orally disintegrating tablets, orally disintegrating tablets They can be safely administered orally or parenterally (e.g., intravenously, intramuscularly, subcutaneously, intraviscerally, intranasally, intradermally, by eye drop, mucosal patch film), injections (e.g., subcutaneous injections, intravenous injections, intramuscular injections, intraperitoneal injections), drops, transdermal preparations, ointments, lotions, patches, suppositories (e.g., rectal suppositories, vaginal suppositories), pellets, nasal preparations, pulmonary preparations (inhalants), eye drops, etc.

As the "pharmaceutical acceptable carrier", various organic or inorganic carriers commonly used as pharmaceutical materials are used. For example, excipients, lubricants, binders, disintegrants, etc. are used in solid preparations, while solvents, solubilizers, suspending agents, isotonicity agents, buffers, soothing agents, etc. are used in liquid preparations. Furthermore, preparation additives such as preservatives, antioxidants, colorants, sweeteners, etc. can also be used as necessary.
Examples of excipients include lactose, sucrose, D-mannitol, starch, corn starch, crystalline cellulose, light anhydrous silicic acid, and the like.
Lubricants include, for example, magnesium stearate, calcium stearate, talc, colloidal silica, and the like.
Examples of binders include crystalline cellulose, sucrose, D-mannitol, dextrin, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, polyvinylpyrrolidone, starch, sucrose, gelatin, methyl cellulose, and sodium carboxymethyl cellulose.
Examples of disintegrants include starch, carboxymethylcellulose, calcium carboxymethylcellulose, sodium carboxymethylstarch, and L-hydroxypropylcellulose.
Examples of the solvent include water for injection, alcohol, propylene glycol, macrogol, sesame oil, corn oil, olive oil, and the like.
Examples of the solubilizing agent include polyethylene glycol, propylene glycol, D-mannitol, benzyl benzoate, ethanol, trisaminomethane, cholesterol, triethanolamine, sodium carbonate, and sodium citrate.
Examples of suspending agents include surfactants such as stearyl triethanolamine, sodium lauryl sulfate, lauryl aminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride, and glycerin monostearate; and hydrophilic polymers such as polyvinyl alcohol, polyvinylpyrrolidone, sodium carboxymethylcellulose, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, and hydroxypropylcellulose.
Examples of the isotonic agent include glucose, D-sorbitol, sodium chloride, glycerin, D-mannitol, and the like.
Examples of the buffering agent include buffer solutions such as phosphate, acetate, carbonate, and citrate.
An example of the soothing agent is benzyl alcohol.
Examples of preservatives include paraoxybenzoic acid esters, chlorobutanol, benzyl alcohol, phenylethyl alcohol, dehydroacetic acid, and sorbic acid.
Examples of antioxidants include sulfites, ascorbic acid, and α-tocopherol.

Pharmaceutical compositions vary depending on the dosage form, administration method, carrier, etc., but can be prepared according to conventional methods by adding compound (I) in a ratio of usually 0.01 to 100% (w/w), preferably 0.1 to 95% (w/w), based on the total amount of the formulation.

The present invention will be explained in more detail below with reference to synthesis examples and test examples, but these do not limit the present invention and may be modified without departing from the scope of the present invention.
In the following examples, "room temperature" generally refers to about 10° C. to about 35° C. Ratios shown in mixed solvents are by volume unless otherwise specified. % refers to weight % unless otherwise specified.

The following compounds used as starting materials are known compounds.
1-Phenyl-1,3,8-triazaspiro[4.5]decan-4-one (CAS Registry Number: 1021-25-6) (Compound 1)
8-Benzyl-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (CAS Registry Number: 974-41-4) (Compound 2)
1-Phenyl-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (CAS Registry Number: 1048-17-5) (Compound 3) (also known as PPTD)

The abbreviations used below are as follows:
MeCN: Acetonitrile MeOH: Methanol THF: Tetrahydrofuran AcOH: Acetic acid n-BuOH: n-butanol DMSO: Dimethylsulfoxide Me ₃ SiCN: Trimethylsilyl cyanide DMF-DMA: N,N-dimethylformamide Dimethylacetal Phe: Phenyl MeO: Methoxy Bn: Benzyl Me: Methyl

Synthesis Example 1
Synthesis of 1-phenyl-8-(3-phenylpropyl)-1,3,8-triazaspiro[4.5]decan-4-one (Compound 4)

To 1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 1) (0.5094 g, 2.20 mmol, 1.0 equiv. in ultra-dehydrated MeCN (40 mL)) was added dry-ground potassium carbonate (0.898 g, 6.5 mmol, 3.0 equiv.), 3-phenylpropyl bromide (0.43 g, 2.16 mmol, 1.0 equiv.), and potassium iodide (0.107 g, 0.648 mmol, 0.3 equiv.). The mixture was refluxed for 2 h, filtered, and the filtrate was collected and concentrated on a rotary evaporator. Using flash column chromatography (Isolera, Sfar HC-Duo 25 g, CHCl ₃ &MeOH: 0%→5%→10%), fractions containing the target compound were collected from the crude product, concentrated, and dried in vacuum to obtain compound 4 (753 mg, 2.15 mmol, yield: 99.5%) as a white solid.

Compounds 5 to 8 shown in Table 1 below were obtained using the same method as in Synthesis Example 1.

Synthesis Example 2
Synthesis of 3-methyl-1-phenyl-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (Compound 9)

To 1-phenyl-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 3, PPTD) (129 mg, 0.384 mmol, 1.0 equiv.) dissolved in THF (9 mL) was added NaH (30.7 mg, 60%, dispersed in paraffin liquid, 0.768 mmol, 2.0 equiv.) in an ice bath (0° C.). The mixture was stirred at 0° C. for 1 hour, and methyl iodide (109 mg, 0.768 mmol, 2.0 equiv.) was added. The mixture was stirred at room temperature overnight (18 hours). The mixture was diluted with ethyl acetate (70 mL) and the reaction was quenched with deionized distilled water (25 mL). The solvent was removed using a rotary evaporator, and the residue was purified using Isolera (Sfar HC Duo 25 g, CHCl ₃ &MeOH: 0%→5%→10%). The fractions containing the target substance were collected and concentrated and dried under reduced pressure to obtain compound 9 (107.7 mg, 0.308 mmol, yield: 80.2%) as a pale yellow oil.

Compounds 10 and 11 shown in Table 1 below were obtained in the same manner as in Synthesis Example 2.

Synthesis Example 3
Synthesis of 8-benzyl-1-(4-methoxyphenyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 12),
Synthesis of 1-(4-methoxyphenyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 13), and
Synthesis of 1-(4-methoxyphenyl)-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (Compound 14)

4-Methoxyaniline (2.45 g, 19.9 mmol) and Me ₃ SiCN (3.0 mL, 24.2 mmol) were added to 1-benzylpiperidin-4-one (4.50 g, 23.8 mmol) stirred in AcOH (25 mL) at 0° C. The reaction mixture was stirred at room temperature for 3.5 hours and quenched with 2N NaOH (225 mL). The solution was extracted with CH ₂ Cl ₂ (50 mL×2), and the organic layer was washed with brine (50 mL×2), dried over MgSO ₄ , filtered, and distilled. Diethyl ether (50 mL) was added to the residue, and the resulting solid was filtered and dried under reduced pressure to give intermediate 1 (4.39 g, 68.7%) as a gray powder.

Intermediate 1 (4.332 g, 13.45 mmol) was added with H ₂ SO ₄ (23 mL) at room temperature and stirred for 22 hours. The reaction was terminated by adding 28% NH ₃ aqueous solution (75 mL) on ice. The solution was extracted with CHCl ₃ (75 mL x 2), the organic layer was washed with brine, dried over Na ₂ SO ₄ , filtered, and the water was evaporated. Diisopropyl ether (25 mL) was added to the residue, and the obtained solid was filtered and dried under reduced pressure to obtain white powder intermediate 2 (3.951 g, 86.5%).

To intermediate 2 (3.842 g, 11.32 mmol) stirred in MeOH (22 mL) was added DMF-DMA (5.0 mL) under nitrogen. The reaction mixture was stirred at 55° C. for 17 h. The solvent was removed under reduced pressure and diisopropyl ether (20 mL) was added to the residue. The resulting solid was filtered and washed with diisopropyl ether and diethyl ether to give intermediate 3 (4.083 g) as a grey powder.

To intermediate 3 (3.93 g, 11.25 mmol) stirred in dehydrated MeOH (160 mL) was added NaBH ₄ (553 mg, 14.6 mmol) under nitrogen. The reaction mixture was stirred at room temperature for 21 hours, and the reaction was quenched by adding water (5 mL). The solvent was removed under reduced pressure, and water (100 mL) was added to the residue. The resulting concentrate was filtered and dried under reduced pressure at 55° C. to obtain compound 12 (3.241 g, 79.7%) as a gray powder.

Compound 12 (3.00 g, 8.536 mmol) stirred in MeOH (265 mL) and AcOH (2.5 mL) was added with 10% Pd/C (600 mg). The reaction mixture was stirred at room temperature under hydrogen for 21 h. After filtration, the residue was evaporated and dissolved in NaHCO ₃ +NaClaq (35 mL). The solution was extracted with n-BuOH (40 mL x 3) and the organic layer was concentrated. The reaction was quenched by adding water (5 mL). The solvent was removed under reduced pressure and diethyl ether (50 mL) was added to the residue. The resulting concentrate was filtered and dried under reduced pressure at 50° C. to give compound 13 (2.21 g, 99.1%) as a white powder.

To a solution of compound 13 (0.2038 g, 0.765 mmol) in ultra-dehydrated MeCN (10 mL) was added dried and ground K ₂ CO ₃ (0.3180 g, 2.304 mmol), 2-phenylethyl bromide (0.1416 g, 0.765 mmol) and potassium iodide (0.0399 g, 0.240 mmol). The mixture was refluxed for 2 h. The mixture was filtered and concentrated on a rotary evaporator. The residue was purified with Isolera (Sfar HC Duo 25 g, CHCl ₃ &MeOH: 0% → 5% → 10%) and the fractions containing the target material were collected and concentrated to dryness under reduced pressure to give compound 14 (270.8 mg, 0.74 mmol, yield: 95%) as a white solid.

Synthesis Example 4
Synthesis of 1-phenyl-8-(2-(4-hydroxy-2-methylphenyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (Compound 39)

To a solution of 1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 1) (179 mg, 0.775 mmol) in dry acetonitrile (15 mL) was added 4-(2-bromoethyl)-3-methylphenol (200 mg, 0.930 mmol) and N,N-diisopropylethylamine (162 μL, 0.930 mmol). The mixture was heated at 70° C. for 24 h, then cooled to room temperature and concentrated under reduced pressure. The residue was purified by flash column chromatography (CHCl ₃ :MeOH=100:3→100:20) to give compound 39 (222 mg, 0.607 mmol, yield: 78%) as a white solid.

Compounds 15 to 20 shown in Table 1 below were obtained using a method similar to that of Synthesis Example 3.

Compounds 21 to 38 shown in Table 1 below were obtained using the same method as in Synthesis Example 1.

The groups in compounds 1 to 39 are shown below.

The NMR data for the compound obtained above is shown below.

Compound 4
¹ H-NMR (400 MHz, CDCl ₃ ): δ/ppm 7.32-7.24 (m, 4H) ^* , 7.24-7.15 (m, 3H), 6.93 (d, J = 8.0 Hz, 2H), 6.87 (t, J = 7.6 Hz), 6.19 (bs, 1H), 4.73 (s, (*: The peak overlapped with the solvent peak (CDCl ₃ ))

Compound 5
¹ H-NMR (400 MHz, METHANOL-D4): δ 7.27 (td, J = 7.0, 2.0 Hz, 2H), 7.14 (dd, J = 6.8, 2.0 Hz, 2H), 7.02 (dd, J = 8.7, 0.8 Hz, 2H), 6.89-6.84 (m, 3H) , 4.70 (s, 2H), 3.76 (s, 3H), 3.16-3.10 (m, 4H), 2.85-2.82 (m, 4H), 2.70-2.62 (m, 2H), 1.82 (d, J = 14.4 Hz, 2H).

Compound 6
¹ H-NMR (400 MHz, METHANOL-D4): δ 7.29-7.23 (m, 4H), 7.04-6.99 (m, 4H), 6.88 (t, J = 7.2 Hz, 1H), 4.70 (s, 2H), 3.16-3.13 (m, 4H), 2.89-2.83 (m, 4H), 2.69-2.61 (m, 2H), 1.82 (d, J = 14.8 Hz, 2H).

Compound 7
¹ H-NMR (400 MHz, METHANOL-D4): δ/ppm 7.29-7.24 (m, 2H), 7.16 (t, J = 7.6 Hz, 1H), 7.05-7.00 (m, 5H), 6.87 (t, J = 7.2 Hz, 1H), 4.70 (s, 2H), 3. 13-3.07 (m, 4H), 2.86-2.80 (m, 4H), 2.70-2.62 (m, 2H), 2.31 (s, 3H), 1.81 (d, J = 14.4 Hz, 2H).

Compound 8
¹ H-NMR (400 MHz, METHANOL-D4): δ 7.30 (t, J = 7.8 Hz, 2H), 7.02 (d, J = 8.4 Hz, 2H), 6.94 (t, J = 7.2 Hz, 1H), 4.72 (s, 2H), 3.62-3.57 (m, 2 H), 3 .41-3.38 (m, 2H), 3.05-3.01 (m, 2H), 2.69-2.61 (m, 2H), 1.99 (d, J = 14.8 Hz, 2H), 1.76-1.66 (m, 5H), 1.62-1.56 (m, 2H), 1.37-1.18 (m, 4H), 1.05-0.95 (m, 2H).

Compound 9
^1H -NMR (400 MHz, CHLOROFORM-D) δ 7.30 (m, 4H), 7.24-7.18 (m, 3H), 6.93 (d, J = 8.0 Hz, 2H), 6.87 (t, J = 7.2 Hz, 1H), 4.68 (s, 2H), 3.00-2.76 (m, 13H) ^# , 1.70 (d, J = 13.2 Hz, 2H). #: The peak at 3.00 (s, 3H) overlapped with these peaks.
ESI-MS: cald. [M+H] ⁺ :350.2227, detection.350.2233., cald. [M+Na] ⁺ :370.2046, detection. 370.2050.

Compound 10
¹ H-NMR (400 MHz, METHANOL-D4): δ 7.29-7.15 (m, 7H), 7.04 (d, J = 8.0 Hz, 2H), 6.86 (t, J = 7.4 Hz, 1H), 4.73 (s, 2H), 3.39 (t, J = 7.2 Hz, 2H), 3 .00-2.91 (m, 4H), 2.87-2.77 (m, 2H), 2.75-2.60 (m, 4H), 1.81-1.61 (m, 4H), 0.96 (t, J = 7.4 Hz, 3H).

Compound 11
¹ H-NMR (400 MHz, METHANOL-D4): δ 7.39-7.16 (m, 12H), 6.97 (d, J = 8.0 Hz, 2H), 6.85 (t, J = 7.4 Hz, 1H), 4.61-4.59 (m, 4H), 3.12-2.92 (m, 4H), 2.88-2.84 (m, 2H), 2.72-2.59 (m, 4H), 1.75 (d, J = 14.4 Hz, 2H).

Compound 14
^1H -NMR (400 MHz, _CDOD3 ): δ/ppm 7.30-7.21 (m, 5H) ^* , 7.05 (d, J = 9.2 Hz, 2H), 6.88 (d, J = 9.0 Hz, 2H), 6.00 (s, 1H), 4.68 (s, 2H), 3.79 (s, 3H), 3.06-2.92 (m, 5H), 1.87 (d, J = 13.7 Hz, 2H), 1.60 (s, 5H). (*: peak overlapped with solvent peak ( _CHCl3 )).
¹ H-NMR (400 MHz, METHANOL-D4): δ/ppm 7.28-7.24 (m, 2H), 7.13 (dd, J = 6.8, 2.2 Hz, 2H), 6.89 (dd, J = 6.8, 2.2 Hz, 2H), 4.63 (s, 2H), 3.76 (s, 3H) ), 3.06-2.97 (m, 4H), 2.84-2.72 (m, 4H), 2.08-2.00 (m, 2H), 1.90 (d, J = 14.5 Hz, 2H).
ESI-MS: cald. [M+H] ⁺ : 366.2176, detected. 366.2180., cald. [M+Na] ⁺ : 388.1995, detected. 388.1999.

Compound 17
¹ H-NMR (400 MHz, CDCl ₃ ): δ/ppm 7.31-7.18 (m, 5H) ^* , 7.03-6.92 (m, 5H), 6.19 (s, 1H), 4.68 (s, 2H), 2.88-2.79 (m, 6H), 2.75-2.66 (m, 2H) , 2.37-2.29 (m, 2H), 1.80 (d, J = 14.3 Hz, 2H). (*: The peak overlapped with the solvent peak (CHCl ₃ ))

Compound 20
¹ H-NMR (400 MHz, CDCl ₃ ): δ/ppm 7.31-7.28 (m, 2H), 7.23-7.19 (m, 3H), 5.92 (brs, 1H), 4.13 (s, 2H), 3.10-2.75 (m, 7H), 2.40 (s, 3H), -2.00 (m, 1H), 1.78-1.53 (m, 4H).

Compound 21
¹ H-NMR (400 MHz, DMSO-D6): δ/ppm 8.63 (s, 1H), 7.23 (t, J = 8.0 Hz, 2H), 7.13 (d, J = 8.4 Hz, 2H), 7.09 (d, J = 8.0 Hz, 2H), 6.82 (d, J = 8.0 Hz, 2H), 6.75 (t, J = 7.2 Hz, 1H), 4.57 (s, 2H), 2.86-2.78 (m, 2H), 2.76-2.67 (m, 4H), 2.59-2.53 (m, 4H) ^* , 2.27 (s, 3H), 1.57 (d, J = 13.2 Hz, 2H). (*: The peak overlapped with the solvent peak (DMSO-D6))

Compound 22
¹ H-NMR (400 MHz, DMSO-D6): δ/ppm 8.64 (s, 1H), 7.23 (t, J = 8.0 Hz, 2H), 7.19-7.06 (m, 4H), 6.84 (d, J = 8.0 Hz, 2H), 6.75 (t, J = 7.2 Hz, 1H), 4.58 (s, 2H), 2.88-2.72 (m, 6H), 2.62-2.52 (m, 4H) ^* , 2.30 (s, 3H), 1.58 (d, J = 13.2 Hz, 2H). (*: The peak overlapped with the solvent peak (DMSO-D6))

Compound 23
¹ H-NMR (400 MHz, CDCl ₃ ): δ/ppm 7.34-7.18 (m, 7H) ^* , 6.87-6.82 (m, 3H), 6.19 (bs, 1H), 4.71 (s, 2H), 3.02-2.92 (m, 1H), 2.84-2.49 (m, 8H) , 1.66 (bd, J = 16.0 Hz, 2H), 1.32 (d, J = 6.8 Hz, 3H). (*: The peak overlapped with the solvent peak (CDCl ₃ ))

Compound 24
¹ H-NMR (400 MHz, CDCl ₃ ): δ/ppm 7.30 (t, J = 8.0 Hz, 2H) ^* , 7.21 (d, J = 8.4 Hz, 2H), 6.96 (dd, J = 6.6Hz, J = 2.0 Hz, 2H), 6.96-6.86 (m, 3H), (bs, 1H), 4.74 (s, 2H), 2.92-2.86 (m, 4H), 2.84-2.78 (m, 2H), 2.72-2.61 (m, 4H), 1.75 (d, J = 14.0 Hz, 2H). (*: The peak overlapped with the solvent peak (CDCl ₃ ))

Compound 25
¹ H-NMR (400 MHz, CDCl ₃ ): δ/ppm 7.59 (dd, J = 8.4 Hz, J = 1.2 Hz), 7.53 (dd, J = 6.4 Hz, J = 2.0 Hz), 7.43 (t, J = 7.6 Hz, 2H), 7.36-7.26 (m, 5H) ^* , 6. 94 (d, J = 8.0 Hz, 2H), 6.88 (t, J = 7.2 Hz, 1H), 6.04 (bs, 1H), 4.74 (s, 2H), 2.96-2.85 (m, 6H), 2.78-2.63 (m, 4H), 1.77 (bd, J = 14.0 Hz, 2 H). (*: The peak overlapped with the solvent peak (CDCl ₃ ))

Compound 26
¹ H-NMR (400 MHz, DMSO-D6): δ/ppm 8.63 (s, 1H), 7.89 (d, J = 7.2 Hz, 1H), 7.85 (dd, J = 8.0 Hz, J = 3.6 Hz, 2H), 7.76 (s, 1H), 7.52-7.42 (m, 3H), 7.18 (t, J = 8.2 Hz, 2H), 6.81 (d, J = 8.0 Hz, 2H), 6.73 (t, J = 7.2 Hz, 1H), 2.98-2.84 (m, 4H), 2.82-2.72 (m, 2H), 2.70-2.63 (m, 2H), 2.6 1-2.54 (m, 3H) ^* , 1.58 (bd, J = 13.2Hz, 2H). (*: The peak overlapped with the solvent peak (DMSO-D6))

Compound 27
¹ H-NMR (400 MHz, CDCl ₃ ): δ/ppm 8.51 (dd, J = 7.6 Hz, J = 1.6 Hz, 2H), 7.29 (t, J = 7.6 Hz, 2H) ^* , 7.16 (d, J = 6.0 Hz, 2H), 6.92-6.85 (m, 3H), 8 (bs, 1H), 4.74 (s, 2H), 2.95-2.79 (m, 6H), 2.75-2.61 (m 4H), 1.75 (bd, J = 14.0 Hz, 2H). (*: The peak overlapped with the solvent peak (CDCl ₃ ))

Compound 28
¹ H-NMR (400 MHz, CDCl ₃ ): δ/ppm 8.50 (d, J = 2.0 Hz, 1H), 8.47 (dd, J = 5.2 Hz, J = 1.6 Hz, 1H), 7.55 (dt, J = 8.0 Hz, J = 2.0 Hz, 1H), 7.29 (t, J = 8.0 Hz, 2H) ^* , 7.23 (dd, J = 7.2 Hz, 4.8 Hz, 1H), 6.94-6.85 (m, 3H), 6.19 (bs, 1H), 4.74 (s, 2H), 2.96-2.86 (m, 4H), 2.86-2.80 (m, 2H), -2.62(m 4H), 1.75 (bd, J = 14.0 Hz, 2H). (*: The peak overlapped with the solvent peak (CDCl ₃ ))

Compound 29
¹ H-NMR (400 MHz, CDCl ₃ ): δ/ppm 8.54 (dq, J = 4.8 Hz, J = 1.6 Hz, J = 0.8 Hz, 1H), 7.61 (dt, J = 7.6 Hz, J = 1.6 Hz, 1H), 7.31-7.27 (m, 2H) ^* , 7.21 7.13 (dt, J = 6.4 Hz, J = 1.2 Hz, 1H), 6.92-6.83 (m 3H), 4.74 (s, 2H), 3.06-3.00 (m 2H), 2.96-2.83 (m 6H), 2.72-2.60 (m , 2H), 1.74 (bd, J = 14.4 Hz, 2H). (*: The peak overlapped with the solvent peak (CDCl ₃ ))

Compound 30
¹ H-NMR (400 MHz, CDCl ₃ ): δ/ppm 7.35-7.24 (m, 10H) ^* , 7.21-7.14 (m, 2H), 6.93 (d, J = 8.0 Hz, 2H), 6.88 (t, J = 7.2 Hz, 1H), 6.05 (bs, 1H), 4. 72 (s, 2H), 4.03 (t, J = 7.6 Hz, 1H), 2.84-2.71 (m, 4H), 2.69-2.57 (m, 2H), 2.41-2.35 (m, 2H), 2.32-2.23 (m, 2H), 1.71 (bd, J = 13.6 Hz, 2H). (*: The peak overlapped with the solvent peak (CDCl ₃ ))

Compound 31
¹ H-NMR (400 MHz, CDCl ₃ ): δ/ppm 7.28 (t, J = 8.0 Hz, 2H) ^* , 7.05 (d, J = 8.0 Hz, 1H), 6.9-6.2 (m, 4H), 6.89 (t, J = 7.2 Hz, 1H), 5.95 (bs, 1H), 4 .72 (s, 2H), 2.95-2.88 (m, 4H), 2.83-2.76 (m, 2H), 2.72-2.58 (m, 4H), 2.29 (s, 3H), 2.28(s, 3H), 1.75 (d, J = 14.4 Hz, 2H). (*: The peak overlapped with the solvent peak (CDCl ₃ ))

Compound 32
¹ H-NMR (400 MHz, CDCl ₃ ): δ/ppm 7.28 (t, J = 8.0 Hz, 2H) ^* , 6.92 (d, J = 7.6 Hz, 2H), 6.87 (t, J = 7.6 Hz, 1H), 6.84 (s, 3H), 6.24 (bs, 1H), (s, 2H), 2.92-2,82 (m, 4H), 2.78-2.62 (m, 6H), 2.28 (s, 6H), 1.75 (bd, J = 14.0 Hz, 2H). (*: The peak overlapped with the solvent peak (CDCl ₃ ))

Compound 33
¹ H-NMR (400 MHz, DMSO-D6): δ/ppm 9.15 (bs, 1H), 8.64 (bs, 1H), 7.23 (t, J = 7.8 Hz, 2H), 7.03 (d, J = 8.0 Hz, 2H), 6.83 (d, J = 8.0 Hz, 2H), (t, J = 7.2 Hz, 1H), 6.67 (d, J = 8.4 Hz, 2H), 4.58 (s, 2H), 2.90-2.40 (m, 10H) ^* , 1.57 (bd, J = 12.0 Hz, 2H). (*: The peak overlapped with the solvent peak (DMSO-D6))

Compound 34
¹ H-NMR (400 MHz, DMSO-D6): δ/ppm 8.63 (s, 1H), 7.74 (d, J = 8.4 Hz, 2H), 7.44 (d, J = 8.4 Hz, 2H), 7.30 (bs, 2H), 7.23 (t, J = 8.0 Hz, 2H), (d, J = 8.0 Hz, 2H), 6.74 (t, J = 7.2 Hz), 4.58 (s, 2H), 2.89-2.79 (m, 4H), 2.78-2.66 (m, 2H), 2.62-2.53 (m,4H) ^* , 1.57 (bd, J = 13.2 Hz, 2 H). (*: The peak overlapped with the Solvent peak (DMSO-D6)

Compound 35
¹ H-NMR (400 MHz, CDCl ₃ ): δ/ppm 7.29 (t, J = 7.6 Hz, 2H) ^* , 7.01 (d, J = 8.4 Hz, 2H), 6.93 (d, J = 8.0 Hz, 2H), 6.88 (t, J = 7.6 Hz, 1H), 6.63 (dd, J = 8.4 Hz, 2H), 6.08 (bs, 1H), 4.74 (s, 2H), 3.49 (bs, 2H), 2.94-2.82 (m, 4H), 2.76-2.60 (m, 6H), 1.75 (bd, J = 14.4 Hz, 2H). (*: The peak overlapped with the solvent peak ( CDCl ₃ ))

Compound 36
¹ H-NMR (400 MHz, CDCl ₃ ): δ/ppm 7.33-7.24 (m, 3H) ^* , 7.01 (d, J = 7.2 Hz, 1H), 6.94-6.86 (m, 5H), 6.13 (bs, 1H), 4.74 (s, 2H), 2.96-2.86 (m, 4H), 2.85-2.78 (m, 2H), 2.73-2.62 (m 4H), 1.75 (bd, J = 14.0 Hz, 2H). (*: The peak overlapped with the solvent peak (CDCl ₃ ))

Compound 37
¹ H-NMR (400 MHz, CDCl ₃ ): δ/ppm 7.33-7.24 (m, 3H) ^* , 7.21 (dd, J = 7.6 Hz, 1.4 Hz, 1H), 7.14 (dd, J = 8.0 Hz, J = 1.2 Hz, 1H), 7.08 (dt, J = 7.2 Hz, 1.2 Hz, 1H), 6.92 (d, J = 8.0 Hz, 2H), 6.88 (t, J = 7.2 Hz, 1H), 6.27 (bs, 1H), 4.74 (s, 2H), 2.94-2.85 (m, 4H), 2.84-2.77 (m, 2H), .60 (m, 4H), 1.75 (bd, J = 14.4 Hz, 2H). (*: The peak overlapped with the solvent peak (CDCl ₃ ))

Compound 38
1H-NMR (400 MHz, CDCl ₃ ): δ/ppm 9.11 (s, 1H), 8.64 (s, 2H), 7.31 (t, J = 8.0 Hz, 2H), 6.91-6.85 (m, 3H), 6.34 (bs, 1H), 4.74 (s, 2H), 2.96-2 .78 (m, 6H), 2.74-2.62 (m 4H), 1.75 (d, J = 14.4 Hz, 2H).

Compound 39
¹ H-NMR (400 MHz, DMSO-d6): δ/ppm 9.03 (s, 1H), 8.63 (s, 1H), 7.24 (t, J = 8.0 Hz, 2H), 6.94 (d, J = 8.4 Hz, 1H), 6.84 (d, ,J = 8.4 Hz, 2H), 6. 75 (t, J = 7.4 Hz, 1H), 6.54 (s, 1H), 6.51 (dd, J = 8.0 Hz, 2.4 Hz, 1H), 4.58 (s, 2H), 3.17 (d, J = 5.2 Hz, 2H), 2.90-2.40 (m, 6H) *, 2.20 (s, 3H), 1.57 (bd, J = 13.6 Hz, 2H) (*: The peak overlapped with the solvent peak (DMSO-D6)
ESI-MS: cald. For [M+H] ⁺ as 366.2176, obsd.366.2167.

Compounds A2 to A30 shown in Table 2 are also specifically exemplified, and these are expected to have the same level of ZIP14 inhibitory activity as the compounds shown in the synthesis examples.

Test Example 1: Evaluation of inhibition of ZIP14-mediated iron transport (using Xenopus oocytes)
cRNA preparation
A DNA fragment of human ZIP14 with an HA tag at the C-terminus was amplified by PCR and cloned using the In-Fusion (registered trademark) HD Cloning Kit (Takara Bio). The PCR primers were designed to contain an HA tag sequence and to have 15 bases in common between both ends of the PCR fragment and the insertion site of the vector. The amplified PCR fragment was inserted into the EcoRV site of the mammalian expression vector pcDNA 3.1(-). Using the above-mentioned plasmid linearized with HindIII as a template, capped cRNA was synthesized using the mMESSAGE mMACHINE T7 Transcription Kit, and poly(A) was added to the cRNA using the poly(A) tailing kit. The poly(A)-tagged cRNA was purified with the MEGAclear Transcription Clean-Up Kit (all Thermo Fisher Scientific) and used for expression and metal uptake experiments using Xenopus oocytes.

Oocyte preparation and transporter expression
Xenopus laevis oocytes were pretreated with 1.5mg/ml collagenase A diluted in OR2 buffer (82mM NaCl, _2.5mM KCl, 1mM MgCl2, 5mM HEPES; pH7.6) for 1.5-2 hours to remove follicles. The oocytes were washed with OR2 buffer and then soaked in ND96 buffer (96mM NaCl, 2mM KCl, 1.8mM _CaCl2 , 1mM _MgCl2 , 5mM HEPES; pH7.5). Oocytes were divided into six stages according to their developmental stage, and the most advanced stages 4 to 6 were used for gene expression experiments. Oocytes at these stages were microinjected with 23 ng/23 nl of cRNA or the same amount of RNA-free water using Nanoject II (Drummond Scientific), and were then incubated at 18°C for 2 days before metal uptake assay.

Metal uptake assay
After injecting cRNA, the oocytes were left to stand for a certain period of time, washed twice with ND96 buffer, and pre-incubated for 30 minutes with ND96 buffer containing the test compound or DMSO. Then, the oocytes were treated with Iron uptake solution (ND 96 buffer containing 10μM FeCl ₂ , 5μCi/ml ⁵⁵ FeCl ₃ , and 1 mM ascorbic acid) containing the test compound (50μM) at room temperature for 30 minutes to allow the oocytes to take up metal ions.
In the iron ion uptake assay, the oocytes were washed five times with ND96 buffer containing 1 mM ascorbic acid, then dissolved in 0.0625 N NaOH, and a scintillation cocktail was added, and the amount of iron ions taken up by the oocytes was measured using a scintillation counter.
In these studies, 3-5 experiments were performed on different days, and the data were normalized to the data from transporter-expressing oocytes treated with DMSO. The normalized data were classified according to the P value (Table 3) and are shown in Table 4.

Table 4 shows that compound (I) inhibits iron transport via ZIP14.

Test Example 2: Evaluation of inhibition of zinc or iron transport via ZIP14 (evaluation of PPTD) (using Xenopus oocytes)
cRNA preparation
A DNA fragment of human ZIP14 with an HA tag at the C-terminus was amplified by PCR and cloned using the In-Fusion (registered trademark) HD Cloning Kit (Takara Bio). The PCR primers were designed to contain an HA tag sequence and to have 15 bases in common between both ends of the PCR fragment and the insertion site of the vector. The amplified PCR fragment was inserted into the EcoRV site of the mammalian expression vector pcDNA 3.1(-). Using the above-mentioned plasmid linearized with HindIII as a template, capped cRNA was synthesized using the mMESSAGE mMACHINE T7 Transcription Kit, and poly(A) was added to the cRNA using the poly(A) tailing kit. The poly(A)-tagged cRNA was purified with the MEGAclear Transcription Clean-Up Kit (all Thermo Fisher Scientific) and used for expression and metal uptake experiments using Xenopus oocytes.

Oocyte preparation and transporter expression
Xenopus laevis oocytes were pretreated with 1.5mg/ml collagenase A diluted in OR2 buffer (82mM NaCl, _2.5mM KCl, 1mM MgCl2, 5mM HEPES; pH7.6) for 1.5-2 hours to remove follicles. The oocytes were washed with OR2 buffer and then soaked in ND96 buffer (96mM NaCl, 2mM KCl, 1.8mM _CaCl2 , 1mM _MgCl2 , 5mM HEPES; pH7.5). Oocytes were divided into six stages according to their developmental stage, and the most advanced stages 4 to 6 were used for gene expression experiments. Oocytes at these stages were microinjected with 23 ng/23 nl of cRNA or the same amount of RNA-free water using Nanoject II (Drummond Scientific), and were then incubated at 18°C for 2 days before metal uptake assay.

Metal uptake assay
After cRNA injection, the oocytes were washed twice with ND96 buffer and pre-incubated with ND96 buffer containing PPTD or DMSO for 30 minutes.Then, oocytes were treated with Iron uptake solution (ND 96 buffer containing 10μM _FeCl2 , 5μCi/ml ^55FeCl3 , and 1mM ascorbic acid) or Zinc uptake solution ₍ ND96 containing 10μM _ZnCl2 and 2μCi/ml ^65ZnCl2 ) containing _PPTD for 30 minutes at room temperature to allow the oocytes to take up metal ions.
In the iron ion uptake assay, the oocytes were washed five times with ND96 buffer containing 1 mM ascorbic acid, then dissolved in 0.0625 N NaOH, and a scintillation cocktail was added, and the amount of iron ions taken up by the oocytes was measured using a scintillation counter.
For the zinc ion uptake assay, zinc-loaded oocytes were washed five times with ND96, exposed to an imaging plate, and visualized with a phosphorimager (Typhoon FLA 9500; GE Healthcare). The images were analyzed using ImageJ to determine the amount of zinc uptake.
In these studies, 3-5 experiments were performed on different days, and the data were normalized to those from transporter-expressing oocytes treated with DMSO. Graphs are shown as mean ± standard deviation.
The results are shown in Figure 1.
FIG. 1 shows that PPTD inhibits the transport of zinc (A) or iron (B) via ZIP14.

Test Example 3: Evaluation of inhibition of manganese or cadmium transport via ZIP14 (evaluation of PPTD) (using TREx-hZIP14 cells)
TREx-hZIP14 cells were seeded in a 6-well plate at ^2x105 cells/well and incubated at 37℃ with 5% _CO2 for 24 hours, after which the medium was replaced with Tet (tetracycline)-containing medium. After incubation for 24 hours, the medium was replaced with serum-free medium, and PPTD (final concentration 0, 10, 50μM), Mn (final concentration 0.1μM) or Cd (final concentration 0.1μM), ^54Mn or ^109Cd were added as tracers and incubated for 1 hour. The cells were washed with serum-containing medium and PBS-EDTA, and then harvested. The intracellular metal concentrations were measured using a gamma counter (AccuFLEXγ7000, HITACHI).
The results are shown in Figure 2.
FIG. 2 shows that PPTD inhibits the transport of manganese (A) or cadmium (B) via ZIP14.

Test Example 4 Evaluation of inhibition of cell damage caused by zinc transport via ZIP14 (evaluation of PPTD) (using TREx-hZIP14 cells)
TREx-hZIP14 cells were seeded at ^1x104 cells/well in a 12-well plate and incubated at 37℃ with 5% _CO2 for 24 hours, after which Tet (tetracycline, final concentration 1μg/mL), _ZnSO4 (final concentration 100μM) and PPTD (final concentration 10μM) were added. For the Tet-free condition, the same amount of ethanol, the solvent in which Tet was dissolved, was added. For the PPTD-free condition, the same amount of DMSO, the solvent in which PPTD was dissolved, was added. After adding the reagents, the cells were left to stand at 37℃ with 5% _CO2 for 72 hours, after which the cell morphology was observed under a phase-contrast microscope and images were taken.
The results are shown in Figure 3.
As can be seen from FIG. 3, PPTD suppresses cell damage caused by zinc transport via ZIP14.

Test Example 5: Evaluation of inhibition of cell damage caused by zinc transport via ZIP14 (evaluation of PPTD) (using C2C12 cells)
Mouse myoblast cell line C2C12 was seeded at ^1x104 cells/well in a 12-well plate and incubated at 37°C and ₅ % CO2 for 24 hours, after which pcDNA5/FRT/TO (Mock, 1μg) or human ZIP14/pcDNA5/FRT/TO plasmid (1μg) was introduced into the cells using Lipofectamine 3000 (registered trademark) (Invitrogen). After standing for 24 hours, each reagent, _ZnSO4 (final concentration 100μM) and PPTD (final concentration 10μM), was added under the conditions shown in the results. For the PPTD-free condition, the same amount of DMSO, the solvent in which PPTD was dissolved, was added. After adding the reagent, the cells were left standing for 5 days under an environment of 37°C and 5% _CO2 , and the morphology of the cells was observed under a phase contrast microscope and images were taken.
The results are shown in Figure 4.
FIG. 4 shows that PPTD suppresses cell damage caused by zinc transport via ZIP14.

Test Example 6: Evaluation of inhibition of cell damage caused by iron transport via ZIP14 (evaluation of PPTD) (using TREx-hZIP14 cells)
TREx-hZIP14 cells were seeded at ^1x104 cells/well in a 12-well plate and incubated at 37℃ with 5% _CO2 for 24 hours. After that, Tet (tetracycline, final concentration 1μg/mL), _FeSO4 (final concentration 20μM) and PPTD (final concentration 10μM) were added under the conditions shown in the results. For the condition without Tet, the same amount of ethanol, the solvent in which Tet was dissolved, was added. For the condition without PPTD, the same amount of DMSO, the solvent in which PPTD was dissolved, was added. After adding the reagents, the cells were left to stand at 37℃ with 5% _CO2 for 72 hours, and then the cell morphology was observed under a phase contrast microscope and images were taken.
The results are shown in Figure 5.
FIG. 5 shows that PPTD suppresses cell damage caused by iron transport via ZIP14.

Test Example 7 Evaluation of inhibition of cell damage caused by manganese or cadmium transport via ZIP14 (evaluation of PPTD) (using TREx-hZIP14 cells)
TREx-hZIP14 cells were seeded in a 6-well plate at ^1x104 cells/well and left to stand for 24 hours at 37℃ with 5% _CO2, after which the medium was replaced with Tet (tetracycline)-containing medium. After 24 hours of standing, PPTD (final concentration 0, 10, 50μM), Mn (final concentration 100μM) or Cd (final concentration 1μM) were added and cultured for 24 hours. After adding alamarBlue (Invitrogen), live and dead cells were counted using a microplate reader to calculate the cell viability.
The results are shown in Figure 6.
FIG. 6 shows that PPTD suppresses cell damage caused by manganese (A) or cadmium (B) transport via ZIP14.

Test Example 8: Evaluation of inhibition of zinc or iron transport via ZIP8 (evaluation of PPTD) (using Xenopus oocytes)
cRNA preparation
A DNA fragment of human ZIP8 with an HA tag at the C-terminus was amplified by PCR and cloned using the In-Fusion (registered trademark) HD Cloning Kit (Takara Bio). The PCR primers were designed to contain an HA tag sequence and to have 15 bases in common between both ends of the PCR fragment and the insertion site of the vector. The amplified PCR fragment was inserted into the EcoRV site of the mammalian expression vector pcDNA 3.1(-). Using the above plasmid linearized with HindIII as a template, capped cRNA was synthesized using the mMESSAGE mMACHINE T7 Transcription Kit, and poly(A) was added to the cRNA using the poly(A) tailing kit. The poly(A)-tagged cRNA was purified with the MEGAclear Transcription Clean-Up Kit (all Thermo Fisher Scientific) and used for expression and metal uptake experiments using Xenopus oocytes.

Oocyte preparation and transporter expression
Xenopus laevis oocytes were pretreated with 1.5mg/ml collagenase A diluted in OR2 buffer (82mM NaCl, _2.5mM KCl, 1mM MgCl2, 5mM HEPES; pgH7.6) for 1.5-2 hours to remove follicles. The oocytes were washed with OR2 buffer and then soaked in ND96 buffer (96mM NaCl, 2mM KCl, 1.8mM _CaCl2 , 1mM _MgCl2 , 5mM HEPES; pH7.5). Oocytes were divided into six stages according to their developmental stage, and the most advanced stages 4 to 6 were used for gene expression experiments. Oocytes at these stages were microinjected with 23 ng/23 nl of cRNA or the same amount of RNA-free water using Nanoject II (Drummond Scientific), and were then incubated at 18°C for 2 days before metal uptake assay.

Metal uptake assay
After cRNA injection, the oocytes were washed twice with ND96 buffer and pre-incubated with ND96 buffer containing PPTD or DMSO for 30 minutes.Then, oocytes were treated with Iron uptake solution (ND 96 buffer containing 10μM _FeCl2 , 5μCi/ml ^55FeCl3 , and 1mM ascorbic acid) or Zinc uptake solution ₍ ND96 containing 10μM _ZnCl2 and 2μCi/ml ^65ZnCl2 ) containing _PPTD for 30 minutes at room temperature to allow the oocytes to take up metal ions.
In the iron ion uptake assay, the oocytes were washed five times with ND96 buffer containing 1 mM ascorbic acid, then dissolved in 0.0625 N NaOH, and a scintillation cocktail was added, and the amount of iron ions taken up by the oocytes was measured using a scintillation counter.
For the zinc ion uptake assay, zinc-loaded oocytes were washed five times with ND96, exposed to an imaging plate, and visualized with a phosphorimager (Typhoon FLA 9500; GE Healthcare). The images were analyzed using ImageJ to determine the amount of zinc uptake.
In these studies, 3-5 experiments were performed on different days, and the data were normalized to those from transporter-expressing oocytes treated with DMSO. Graphs are shown as mean ± standard deviation.
The results are shown in Figure 7.
FIG. 7 shows that PPTD does not inhibit the transport of zinc (A) or iron (B) via ZIP8.

Test Example 9: Evaluation of inhibition of manganese or cadmium transport via ZIP8 (evaluation of PPTD) (using TREx-hZIP8 cells)
TREx-hZIP8 cells were seeded in a 6-well plate at ^2x105 cells/well and incubated at 37℃ with 5% _CO2 for 24 hours, after which the medium was replaced with Tet (tetracycline)-containing medium. After incubation for 24 hours, the medium was replaced with serum-free medium, and PPTD (final concentration 0, 10, 50μM), Mn (final concentration 0.1μM) or Cd (final concentration 0.1μM), ^54Mn or ^109Cd were added as tracers and incubated for 1 hour. The cells were washed with serum-containing medium and PBS-EDTA, and then harvested. The intracellular metal concentrations were measured using a gamma counter (AccuFLEXγ7000, HITACHI).
The results are shown in Figure 8.
FIG. 8 shows that PPTD does not inhibit the transport of manganese (A) or cadmium (B) via ZIP8.

Test Example 10: Evaluation of inhibition of cell damage caused by zinc transport via ZIP8 (evaluation of PPTD) (using TREx-hZIP8 cells)
TREx-hZIP8 cells were seeded at ^1x104 cells/well in a 12-well plate and incubated at 37℃ with 5% _CO2 for 24 hours, after which Tet (tetracycline, final concentration 1μg/mL), _ZnSO4 (final concentration 100μM) and PPTD (final concentration 10μM) were added. For the Tet-free condition, the same amount of ethanol, the solvent in which Tet was dissolved, was added. For the PPTD-free condition, the same amount of DMSO, the solvent in which PPTD was dissolved, was added. After adding the reagents, the cells were left to stand at 37℃ with 5% _CO2 for 72 hours, after which the cell morphology was observed under a phase-contrast microscope and images were taken.
The results are shown in Figure 9.
FIG. 9 shows that PPTD does not suppress cell damage caused by zinc transport via ZIP8.

Test Example 11: Evaluation of inhibition of cell damage caused by iron transport via ZIP8 (evaluation of PPTD) (using TREx-hZIP8 cells)
TREx-hZIP8 cells were seeded at ^1x104 cells/well in a 12-well plate and incubated at 37℃ with 5% _CO2 for 24 hours. After that, Tet (tetracycline, final concentration 1μg/mL), _FeSO4 (final concentration 20μM) and PPTD (final concentration 10μM) were added under the conditions shown in the results. For the Tet-free condition, the same amount of ethanol, the solvent in which Tet was dissolved, was added. For the PPTD-free condition, the same amount of DMSO, the solvent in which PPTD was dissolved, was added. After adding the reagents, the cells were left to stand at 37℃ with 5% _CO2 for 72 hours, and then the cell morphology was observed under a phase-contrast microscope and images were taken.
The results are shown in Figure 10.
FIG. 10 shows that PPTD does not suppress cell damage caused by iron transport via ZIP8.

Test Example 12: Evaluation of inhibition of cell damage caused by manganese or cadmium transport via ZIP8 (evaluation of PPTD) (using TREx-hZIP8 cells)
TREx-hZIP8 cells were seeded in a 6-well plate at ^1x104 cells/well and left to stand for 24 hours at 37℃ with 5% _CO2, after which the medium was replaced with Tet (tetracycline)-containing medium. After 24 hours of standing, PPTD (final concentration 0, 10, 50μM), Mn (final concentration 100μM) or Cd (final concentration 1μM) were added and cultured for 24 hours. After adding alamarBlue (Invitrogen), live and dead cells were counted using a microplate reader to calculate the cell viability.
The results are shown in Figure 11.
FIG. 11 shows that PPTD does not suppress cell damage caused by manganese (A) or cadmium (B) transport via ZIP8.

Test Example 13: Evaluation of inhibition of proliferation of hepatic cancer cells (evaluation of PPTD) (use of HepG2 cell line)
Assay method: For the culture of the HepG2 cell line, a human hepatoma-derived cell line, D-MEM medium (Fujifilm, #043-30085) containing fetal bovine serum (final concentration 10%) and antibiotics (Nacalai, #09366-44) was used. The HepG2 cell line was seeded at ^2x104 cells per well in a 12-well plate and cultured at 37°C and 5% _CO2 . 24 hours after cell seeding, PPTD solution (final concentration 10mM) was added. As a negative control, DMSO solution (final concentration 0.1%), which is the solvent for the PPTD solution, was added. Three days after the addition of PPTD, the medium in the well was removed and washed with PBS(-), and then 300mL of trypsin-EDTA solution (Nacalai, #32777-15) was added and incubated for 5 minutes. After confirming that the cells had detached from the bottom of the well using a phase contrast microscope, 1 mL of the above-mentioned D-MEM medium was added and a cell suspension was prepared by pipetting. A portion of the cell suspension was taken and mixed with an equal amount of trypan blue to stain dead cells, and the number of live cells was counted using a hemocytometer.
The results are shown in Figure 12.
12, PPTD exhibited a significant cell proliferation inhibitory effect on HepG2, suggesting that PPTD inhibits the proliferation of hepatic cancer cells by inhibiting ZIP14.

Test Example 14 Solubility test of PPTD and its hydrochloride
Preparation of Compound 3 Hydrochloride (PPTD-HCl)
PPTD (166.6 mg, 0.497 mmol) was dissolved in ethyl acetate (approximately 20 mL), and 1 mL (1 mmol) of a 1M diethyl ether solution of hydrogen chloride was added to confirm that the solution became cloudy. After stirring for approximately 1 hour, the solution was filtered, and the solid on the filter paper was washed several times with ethyl acetate and then dried to obtain PPTD-HCl as a white solid (133.6 mg, 0.359 mmol).
The structure of the product was confirmed to be the following structure by ¹ H NMR (1.0 mg/0.5 mL in D ₂ O: deuterium oxide).

Solubility Test
We tried to dissolve PPTD (1 mg) in 0.5 mL of D ₂ O (heavy water), but no dissolution was observed visually. We measured the 1H NMR spectrum in D ₂ O, but no signal from the compound was observed, so we believe that PPTD is hardly dissolved under these conditions.
On the other hand, PPTD-HCl was completely dissolved under the above conditions, and the 1H NMR spectrum was also observable, so it was concluded that the solubility of PPTD-HCl was significantly improved compared to PPTD.

Test Example 15: Evaluation of inhibition of iron transport via ZIP14 (using Xenopus oocytes)
Two days after injecting human ZIP14 cRNA into Xenopus oocytes in the same manner as in Test Example 1, the cells were incubated for 30 minutes in ND96 buffer containing each compound. The buffer was then replaced with ND96 buffer containing ⁵⁵ Fe and each compound, and the cells were incubated for 30 minutes. The oocytes were washed with ND96 buffer not containing ⁵⁵ Fe, and the radioactivity of ⁵⁵ Fe transported into the cells was measured.
The results are shown in Figure 13.
FIG. 13 shows that PPTD-HCl inhibits iron transport mediated by ZIP14 to the same extent as PPTD.

Test Example 16: Evaluation of irreversible inhibition of iron transport via ZIP14 (evaluation of PPTD) (using Xenopus oocytes)
Human ZIP14cRNA was injected into Xenopus oocytes in the same manner as in Test Example 1, and after culturing for 2 days, the cells were left to stand in ND96 buffer containing the compound or DMSO for 30 minutes. The oocytes were then washed in ND96 buffer, and after the specified time, the buffer was replaced with ND96 buffer containing ^55Fe . After 10 minutes, the buffer was replaced with ND96 buffer not containing ^55Fe , and the oocytes were washed. The oocytes were collected, and the radioactivity of ^55Fe transported into the cells was measured. The relative radioactivity value with DMSO treatment at each time is also shown.
The radioactivity of ⁵⁵ Fe is shown in FIG. 14, and the relative values are shown in FIG.
14 and 15, Fe uptake was observed even after washing the oocytes for 10 minutes, indicating that the inhibition of iron transport mediated by ZIP14 by PPTD is irreversible.

Test Example 17: Evaluation of inhibition of zinc transport via ZIP14 (IC ₅₀ ) (using Xenopus oocytes)
Human ZIP14 cRNA was injected into Xenopus oocytes in the same manner as in Test Example 1, and after 2 days, the cells were placed in ND96 buffer containing 0.03 to 50 μM of each compound for 30 minutes. The buffer was then replaced with ND96 buffer containing ⁶⁵ Zn and 0.03 to 50 μM of each compound, and the cells were placed in ND96 buffer for 30 minutes. The oocytes were then washed with ND96 buffer containing 5 mM EDTA, and the radioactivity of ⁶⁵ Zn transported into the cells was measured. _IC50 was calculated from a four-coefficient logistic regression curve.
The results are shown in Table 5.

Table 5 shows that compound 39 inhibits zinc transport via ZIP14 to the same extent as PPTD.

Test Example 18: Evaluation of inhibition of iron transport via ZIP14 (IC ₅₀ ) (using Xenopus oocytes)
Human ZIP14 cRNA was injected into Xenopus oocytes in the same manner as in Test Example 1, and after 2 days, the cells were placed in ND96 buffer containing 0.03 to 50 μM of each compound, and then the buffer was replaced with ND96 buffer containing ⁵⁵ Fe, 1 mM ascorbic acid, and 0.03 to 50 μM of each compound. After leaving the cells for 30 minutes, the buffer was replaced with ND96 buffer containing 1 mM ascorbic acid, and the radioactivity of ⁵⁵ Fe transported into the cells was measured. _IC50 was calculated from a four-coefficient logistic regression curve.
The results are shown in Table 6.

Table 6 shows that compounds 37 and 39 inhibit ZIP14-mediated iron transport to the same extent as PPTD.

Test Example 19: Evaluation of ZIP14-mediated inhibition of reactive oxygen species (ROS) production (evaluation of PPTD) (using TREx-hZIP14 cells)
TREx-hZIP14 cell line was seeded in a 96-well plate at 8x10 ³ cells/well, and cultured for 24 hours under conditions of 37°C and 5% CO ₂ concentration. Then, tetracycline (Tet: final concentration 1μg/mL) and PPTD (final concentration 10μM) were added according to the conditions shown in FIG. 16. In the Tet-free condition (Tet(-)), the same amount of DMSO used to dissolve Tet was added. Similarly, in the PPTD-free condition, the same amount of DMSO used to dissolve PPTD was added. After adding the above reagents, the cells were cultured for 72 hours under conditions of 37°C and 5% CO ₂ concentration, and then ROS detection reagent (Cell ROX ^TM Green Reagent, Invitrogen) was added and the cells were left to stand for 30 minutes under conditions of 37°C and 5% CO ₂ concentration. Then, the cell morphology and the fluorescence derived from the ROS detection reagent were observed under a fluorescent microscope, and image data were obtained. The fluorescence intensity derived from the ROS detection reagent in each sample was quantitatively evaluated by image analysis using ImageJ. The results are shown in Figure 16.
As shown in FIG. 16, the production of ROS dependent on the expression of ZIP14 by addition of Tet was significantly suppressed by the addition of PPTD.

Test Example 20: Evaluation of inhibition of lipid peroxide (LPO) production via ZIP14 (evaluation of PPTD) (using TREx-hZIP14 cells)
TREx-hZIP14 cell line was seeded in a 96-well plate at 8x10 ³ cells/well, and cultured for 24 hours under conditions of 37°C and 5% CO ₂ concentration, after which tetracycline (Tet: final concentration 1μg/mL) and PPTD (final concentration 10μM) were added according to the conditions shown in FIG. 17. In the Tet-free condition (Tet(-)), the same amount of DMSO used to dissolve Tet was added. Similarly, in the PPTD-free condition, the same amount of DMSO used to dissolve PPTD was added. After adding the above reagents, the cells were cultured for 72 hours under conditions of 37°C and 5% CO ₂ concentration, and then LPO detection reagent (Liperfluo ^TM , Dojindo) was added and left to stand for 30 minutes under conditions of 37°C and 5% CO ₂ concentration. Then, the cell morphology and the fluorescence derived from the LPO detection reagent were observed under a fluorescent microscope, and image data were obtained. The intensity of the fluorescence from the LPO detection reagent in each sample was quantitatively evaluated using FACS, and the results are shown in Figure 17.
As shown in FIG. 17, the production of LPO dependent on the expression of ZIP14 by the addition of Tet was significantly suppressed by the addition of PPTD.

The following formulations are examples of formulations of the present invention. However, the present invention is not limited to these formulation examples.

Formulation Example 1: Preparation of capsules 1) Compound 3 (PPTD) 30 mg
2) Microcrystalline cellulose 10 mg
3) Lactose 19mg
4) Magnesium stearate 1 mg
1), 2), 3) and 4) are mixed and filled into a gelatin capsule.

Formulation Example 2: Preparation of tablets 1) Compound 3 (PPTD) 10 g
2) Lactose 50g
3) 15g corn starch
4) Carmellose calcium 44g
5) Magnesium stearate 1g
The total amount of 1), 2), and 3) and 30 g of 4) are mixed with water, vacuum dried, and sized. 14 g of 4) and 1 g of 5) are mixed with the sized powder, and the mixture is compressed into tablets using a tablet press. In this way, 1000 tablets containing 10 mg of compound 3 (PPTD) per tablet are obtained.

According to the present invention, compound (I) has an inhibitory effect on the metal transporter ZIP14, and is therefore expected to be useful in the prevention or treatment of diseases associated with ZIP14 (e.g., cancer cachexia accompanied by skeletal muscle atrophy or loss, iron overload (including liver damage associated with iron overload), liver cancer, manganese or cadmium poisoning, etc., particularly cancer cachexia accompanied by skeletal muscle atrophy or loss).
Furthermore, compound (I) does not affect metal transport by the metal transporter ZIP8, which is most closely related to the metal transporter ZIP14 in terms of molecular evolution, and therefore serves as a selective inhibitor of the metal transporter ZIP14.

Claims (17)

Formula (I):
[Wherein,
R ¹ represents a C _6-10 aryl group, a C _7-14 aralkyl group, a 5- or 6-membered aromatic heterocyclic group, a 3- to 8-membered non-aromatic heterocyclic group, a C _3-8 cycloalkyl group, a C _3-8 cycloalkenyl group, or a C _1-6 alkyl group, in which the C _6-10 aryl group, C _7-14 aralkyl group, a 5- or 6-membered aromatic heterocyclic group, a 3- to 8-membered non-aromatic heterocyclic group, a C _3-8 cycloalkyl group, or a C _3-8 cycloalkenyl group is optionally substituted with a substituent selected from the substituent group Z;
R ² represents a hydrogen atom or a C _1-6 alkyl group;
R ³ represents a hydrogen atom, a C _1-6 alkyl group, a C _6-10 aryl group, a C _7-14 aralkyl group, a 5- or 6-membered aromatic heterocyclic group, a 3- to 8-membered non-aromatic heterocyclic group, a C _3-8 cycloalkyl group, or a C _3-8 cycloalkenyl group, in which the C _6-10 aryl group, C _7-14 aralkyl group, a 5- or 6-membered aromatic heterocyclic group, a 3- to 8-membered non-aromatic heterocyclic group, a C _3-8 cycloalkyl group, or a C _3-8 cycloalkenyl group is optionally substituted by a substituent selected from the substituent group Z;
R ⁴ is selected from a halogen atom, a cyano group, a nitro group, a C _1-6 alkyl group, a C _1-6 haloalkyl group, a hydroxy group, a C _1-6 alkoxy group, a C _1-6 haloalkoxy group, an amino group, a mono-C _1-6 alkylamino group, a di-C _1-6 alkylamino group, a carboxy group, a C _1-6 alkyl-carbonyl group, and a C _1-6 alkoxy-carbonyl group;
n represents an integer of 0 to 2;
^R5 is a hydrogen atom, a _C1-6 alkyl group, or a group represented by the formula: -L-Cy
(Wherein,
L is a bond, a C _1-6 alkylene group, or a group of the formula: *-(CH ₂ ) _m1 -X-(CH ₂ ) _m2 -**, where:
X represents O, S, NR ⁶ , CO, NR ⁶ CO, CONR ⁶ , COO, or OCO, R ⁶ represents a hydrogen atom or a C _1-6 alkyl group;
m1 represents an integer of 0 to 3;
m2 represents an integer of 0 to 3;
* indicates the bonding site with the nitrogen atom of the spiro ring.
** indicates the binding site with Cy.)
represents a group represented by
Cy represents a C _6-10 aryl group, a 5- or 6-membered aromatic heterocyclic group, a 3- to 8-membered non-aromatic heterocyclic group, a C _3-8 cycloalkyl group, or a C _3-8 cycloalkenyl group, in which the C _6-10 aryl group, the 5- or 6-membered aromatic heterocyclic group, the 3- to 8-membered non-aromatic heterocyclic group, the C _3-8 cycloalkyl group, or the C _3-8 cycloalkenyl group may be substituted with a substituent selected from the substituent group Z.
represents a group represented by
Substituent group Z consists of a halogen atom, a cyano group, a nitro group, a C _1-6 alkyl group, a C _1-6 haloalkyl group, a hydroxy group, a C _1-6 alkoxy group, a C _1-6 haloalkoxy group, an amino group, a mono-C _1-6 alkylamino group, a di-C _1-6 alkylamino group, a carboxy group, a C _1-6 alkyl-carbonyl group, a C _1-6 alkoxy-carbonyl group, an azido group, a carbamoyl group, a mono-C _1-6 alkyl-carbamoyl group, a di-C _1-6 alkyl-carbamoyl group, a sulfamoyl group, a mono-C _{1-6 alkyl-sulfamoyl group, a di-C 1-6 alkyl} -sulfamoyl group, a sulfo group, a methylenedioxy group (-OCH ₂ O-), and a C _6-10 aryl group.]
or a salt thereof. The inhibitor according to claim 1, wherein R ¹ is a C _6-10 aryl group, a C _7-14 aralkyl group, a 5- or 6-membered aromatic heterocyclic group, a 3- to 8-membered non-aromatic heterocyclic group, a C _3-8 cycloalkyl group, or a C _3-8 cycloalkenyl group, wherein the C _6-10 aryl group, a C _7-14 aralkyl group, a 5- or 6-membered aromatic heterocyclic group, a 3- to 8-membered non-aromatic heterocyclic group, a C _3-8 cycloalkyl group, or a C _3-8 cycloalkenyl group is optionally substituted with a substituent selected from substituent group Z. The inhibitor according to claim 1 , wherein R ² is a hydrogen atom. The inhibitor according to claim 1, wherein R ³ is a hydrogen atom or a C _1-2 alkyl group. The inhibitor of claim 1, wherein n is 0. ^R5 is a group of the formula: -L-Cy;
L is a C _1-6 alkylene group; and
Cy has the same meaning as in claim 1.
The inhibitor according to claim 1. The compound represented by formula (I) or a salt thereof
1-phenyl-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 3) or a salt thereof;
The inhibitor according to claim 1, which is 8-(2-(2-azidophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 37) or a salt thereof; or 1-phenyl-8-(2-(4-hydroxy-2-methylphenyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 39) or a salt thereof. The inhibitor according to claim 1, wherein the compound represented by formula (I) or a salt thereof is 1-phenyl-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one or its hydrochloride salt. The inhibitor according to claim 1, which inhibits transport of zinc, iron, manganese or cadmium via the metal transporter ZIP14. The inhibitor according to claim 1, which suppresses cell damage caused by the transport of zinc, iron, manganese or cadmium via the metal transporter ZIP14. The inhibitor according to claim 1, which does not inhibit the metal transporter ZIP8. The inhibitor according to claim 11, which does not inhibit the transport of zinc, iron, manganese or cadmium via the metal transporter ZIP8 and does not suppress cell damage caused by said transport. A preventive or therapeutic agent for a disease associated with the metal transporter ZIP14, comprising a compound defined in any one of claims 1 to 12 or a salt thereof. The preventive or therapeutic agent according to claim 13, wherein the disease associated with the metal transporter ZIP14 is cancer cachexia accompanied by atrophy or loss of skeletal muscle, iron overload, liver cancer, or manganese or cadmium toxicity. A pharmaceutical composition for inhibiting the metal transporter ZIP14, comprising a compound defined in any one of claims 1 to 12 or a salt thereof, and a pharma- ceutical acceptable carrier. A pharmaceutical composition for preventing or treating a disease associated with the metal transporter ZIP14, comprising a compound defined in any one of claims 1 to 12 or a salt thereof, and a pharma- ceutical acceptable carrier. 8-(2-(3-methylphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 7);
3-Methyl-1-phenyl-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 9);
1-Phenyl-8-(2-phenylethyl)-3-propyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 10);
3-benzyl-1-phenyl-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 11);
1-(4-methoxyphenyl)-8-(2-phenylethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 14);
8-(2-(4-azidophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 24);
8-(2-(4-biphenylyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 25);
8-(2-(2-naphthyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 26);
8-(2-(2,4-dimethylphenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 31);
1-Phenyl-8-(2-(4-sulfamoylphenyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 34);
8-(2-(3-azidophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 36);
8-(2-(2-azidophenyl)ethyl)-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (compound 37);
1-Phenyl-8-(2-(5-pyrimidinyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 38); and 1-Phenyl-8-(2-(4-hydroxy-2-methylphenyl)ethyl)-1,3,8-triazaspiro[4.5]decan-4-one (compound 39).
or a salt thereof.