JP7237311B2 - Compounds, pharmaceutical compositions, KDM5C inhibitors and antidepressants - Google Patents

Description

The present invention relates to compounds, pharmaceutical compositions, KDM5C inhibitors and antidepressants.

KDM5 is known to be associated with cancer. For example, Patent Document 1 discloses that a KDM5 antagonist is effective in cancer treatment and prevention of drug resistance in cancer. In addition, Patent Document 2,
It discloses that compounds capable of modulating the activity of histone demethylase are effective in treating cancer.

Japanese Patent Publication No. 2016-520528 Japanese translation of PCT publication No. 2017-512804

An object of the present invention is to provide compounds, pharmaceutical compositions, and antidepressants having potent KDM5C inhibitory activity.

The present invention provides the following compounds, pharmaceutical compositions, KDM5C inhibitors and antidepressants.
Section 1. Formula (I) below

(wherein any one of R ¹ to R ⁴ represents COR ⁸ , and the other three of R ¹ to R ⁴ are the same or different, hydrogen atom, halogen atom, OH, NO ₂ , CN, alkyl, cycloalkyl, alkoxy, alkylcarbonylamino, alkylcarbonyloxy, arylcarbonyloxy, carbamoyl, arylcarbonylamino, aryl, aralkyl, alkylcarbonyl, SH or alkylthio R ¹ and R ² , R ² and R ³ , R ³ and R ⁴ represent methylenedioxy, or together with the carbon atoms to which they are attached, a cyclopentene ring, cyclopentadiene ring, cyclohexene ring, cyclohexadiene ring, benzene ring, or a 5-membered or A 6-membered heterocycle may be formed.
^R5 represents a hydrogen atom, a halogen atom, alkyl, alkoxy, aryl, aralkyl, hydroxyalkyl or cycloalkyl. R ⁴ and R ⁵ together with the carbon atom to which they are bonded may form an optionally substituted benzene ring or an optionally substituted pyridine ring.
R ⁶ and R ⁷ are the same or different and represent a hydrogen atom, alkyl, alkoxy, aryl, aralkyl, hydroxyalkyl or cycloalkyl. However, ^R6 and ^R7 are not hydrogen atoms at the same time.
R ⁸ represents OH, alkoxy, hydroxyalkyloxy, cycloalkyloxy, aryloxy or aralkyloxy.
Z represents N or ^CR9 . ^R9 represents a hydrogen atom, alkyl, aryl or aralkyl.
n represents an integer of 0-5. )
A compound represented by or a pharmaceutically acceptable salt or solvate thereof.
Section 2. A pharmaceutical composition comprising the compound of Item 1 or a pharmaceutically acceptable salt or solvate thereof and a pharmaceutically acceptable excipient.
Item 3. A KDM5C inhibitor comprising the compound according to item 1 or a pharmaceutically acceptable salt or solvate thereof as an active ingredient.
Section 4. Item 2. An antidepressant comprising the compound of item 1 or a pharmaceutically acceptable salt or solvate thereof as an active ingredient.

ADVANTAGE OF THE INVENTION According to this invention, the antidepressant which has a strong KDM5C inhibitory effect can be provided.

KDM inhibitory activity of NPC-3422, CPI-455 and NCDM-81a KDM5C selectivity in cells GxE depression model: smSDS model Anti-stress effects of KDM5C inhibitors Effect of KDM5C inhibitor in depression model (sucrose preference test). Search for KDM5C target genes involved in depression-like behavior. RNA-seq was used to analyze changes in brain gene expression in three groups: no stress, stress + vehicle, and stress + KDM5C inhibitor (n=4 for each group). (Left) Principal component analysis. A method for integrating multivariate data and creating a new comprehensive index. Data summarized from multi-dimensional data into 1-3 dimensions in an easy-to-understand manner. The degree of similarity between groups is indicated by distance, and if the distance is large, genes with significant differences between groups can be extracted. (Right) Clustering analysis. It was shown that the degree of similarity (gene expression pattern) between "non-stress" and "stress + KDM5C inhibitor" is high. That is, it indicates that the abnormal gene expression pattern caused by stress load is normalized by the KDM5C inhibitor.

The compounds of the present invention have the following formula (I)

(Wherein, R ¹ to R ⁷ , Z, and n are as defined above.)
is a compound represented by or a pharmaceutically acceptable salt or solvate thereof.

In one preferred embodiment of the present invention, R ³ represents COR ⁸ , R ¹ , R ² , R ⁴ are the same or different, hydrogen atom, halogen atom, OH, NO ₂ , CN, alkyl, cycloalkyl, Alkoxy, alkylcarbonyloxy, arylcarbonyloxy, carbamoyl, alkylcarbonylamino, aryl, aralkyl, alkylcarbonyl, SH or alkylthio. R ¹ and R ² represent methylenedioxy, or together with the carbon atoms to which they are attached, a cyclopentene ring, cyclopentadiene ring, cyclohexene ring, cyclohexadiene ring, benzene ring, or a 5-membered or A 6-membered heterocycle may be formed.

In another preferred embodiment of the present invention, R ² represents COR ⁸ and R ¹ , R ³ , R ⁴ are the same or different and represent a hydrogen atom, a halogen atom, OH, NO ₂ , CN, alkyl, cyclo represents alkyl, alkoxy, alkylcarbonyloxy, arylcarbonyloxy, carbamoyl, alkylcarbonylamino, aryl, aralkyl, alkylcarbonyl, SH or alkylthio. R ³ and R ⁴ represent methylenedioxy, or together with the carbon atoms to which they are attached, a cyclopentene ring, cyclopentadiene ring, cyclohexene ring, cyclohexadiene ring, benzene ring, or a 5-membered or A 6-membered heterocycle may be formed.

In another preferred embodiment of the present invention, R ¹ represents COR ⁸ , R ² to R ⁴ are the same or different, hydrogen atom, halogen atom, OH, NO ₂ , CN, alkyl, cycloalkyl, alkoxy , alkylcarbonyloxy, arylcarbonyloxy, carbamoyl, alkylcarbonylamino, aryl, aralkyl, alkylcarbonyl, SH or alkylthio. R ² and R ³ , R ³ and R ⁴ each represent methylenedioxy, or together with the carbon atoms to which they are attached, cyclopentene ring, cyclopentadiene ring, cyclohexene ring, cyclohexadiene ring, benzene A ring or a 5- or 6-membered heterocyclic ring may be formed.

In another preferred embodiment of the invention ^R5 is a hydrogen atom, a fluorine atom or alkyl.

In another preferred embodiment of the invention, R ⁶ , R ⁷ are the same or different and represent a hydrogen atom, alkyl, aralkyl, hydroxyalkyl or cycloalkyl, more preferably alkyl.

In another preferred embodiment of the invention ^R8 is OH, alkoxy, aryloxy or aralkyloxy.

In another preferred embodiment of the invention ^R9 is a hydrogen atom or alkyl.

In another preferred embodiment of the present invention, preferred compounds of the present invention represented by general formula (I) are those in which R ³ represents COR ⁸ and R ¹ , R ² , R ⁴ , R ⁵ are all It is a hydrogen atom.

n is an integer of 0 to 5, preferably an integer of 0 to 4, more preferably an integer of 1 to 3, more preferably 1 or 2, particularly preferably 1;

Z is N (nitrogen atom) or CR ⁹ , more preferably N.

Solvents constituting solvates include water, alcohols (methanol, ethanol, n-propanol, isopropanol, butanol, etc.), acetone, methyl ethyl ketone, ethyl acetate, tetrahydrofuran, diethyl ether, diisopropyl ether, benzene, toluene, methylene chloride, chloroform, acetonitrile, dimethylformamide, dimethylsulfoxide and the like.

Pharmaceutically acceptable salts include alkali metal salts such as sodium salts, potassium salts, lithium salts, hydrofluorates, hydrochlorides, hydrobromides, hydroiodide, sulfates, nitrates, phosphates salts, inorganic acid salts such as perchlorates, methanesulfonates, ethanesulfonates, benzenesulfonates, toluenesulfonates, acetates, lactates, maleates, fumarates, succinates, Organic acid salts such as citrate, pyruvate and benzoate are included.

In the present specification, the halogen atom includes a fluorine atom, a chlorine atom, a bromine atom and an iodine atom, preferably a fluorine atom, a chlorine atom and a bromine atom, more preferably a fluorine atom and a chlorine atom.

Alkyl includes linear or branched chain such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, hexyl, heptyl, octyl, 2-ethylhexyl, nonyl and decyl. C _1-10 alkyl, preferably C _1-8 alkyl, more preferably C _1-6 alkyl having

Alkylthio includes linear or branched C _1-6 alkylthio such as methylthio, ethylthio, n-propylthio, isopropylthio, n-butylthio, isobutylthio, tert-butylthio, n-pentylthio, isopentylthio and hexylthio. mentioned.

Hydroxyalkyl includes C _1-6 hydroxyalkyl such as hydroxymethyl, 2-hydroxyethyl, 3-hydroxypropyl, 4-hydroxybutyl, 5-hydroxypentyl, 6-hydroxyhexyl and the like.

Hydroxyalkyloxy includes C _1-6 hydroxyalkyloxy such as hydroxymethyloxy, 2-hydroxyethyloxy, 3-hydroxypropyloxy, 4-hydroxybutyloxy, 5-hydroxypentyloxy and 6-hydroxyhexyloxy. be done.

Cycloalkyl includes C _3-7 cycloalkyl such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl.

Cycloalkyloxy includes C _3-7 cycloalkyloxy such as cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy and cycloheptyloxy.

Alkylcarbonylamino includes methylcarbonylamino, ethylcarbonylamino, n-propylcarbonylamino, isopropylcarbonylamino, n-butylcarbonylamino, isobutylcarbonylamino, tert-butylcarbonylamino, n-pentylcarbonylamino and isopentylcarbonylamino. , straight or branched C _1-6 alkylcarbonylamino such as hexylcarbonylamino.

Alkylcarbonyl includes linear or branched chain such as methylcarbonyl, ethylcarbonyl, n-propylcarbonyl, isopropylcarbonyl, n-butylcarbonyl, isobutylcarbonyl, tert-butylcarbonyl, n-pentylcarbonyl, isopentylcarbonyl, hexylcarbonyl, etc. C _1-6 alkylcarbonyl having.

Aryl means a monocyclic or polycyclic group consisting of a 5- or 6-membered aromatic hydrocarbon ring, specifically phenyl, naphthyl, fluorenyl, anthryl, biphenylyl, tetrahydronaphthyl, chromanyl, 2, 3-dihydro-1,4-dioxanaphthalenyl, indanyl and phenanthryl.

5- or 6-membered heterocycles include furan, thiophene, pyrrole, imidazole, pyrazole, oxazole, thiazole, isoxazole, isothiazole, pyridine, pyrazine, pyrimidine, pyridazine.

Specific examples of alkylcarbonyloxy include methylcarbonyloxy, ethylcarbonyloxy, n-propylcarbonyloxy, isopropylcarbonyloxy, n-butylcarbonyloxy, isobutylcarbonyloxy, tert-butylcarbonyloxy, n-pentylcarbonyloxy, iso Examples include C _1-6 alkylcarbonyloxy such as pentylcarbonyloxy, hexylcarbonyloxy and the like.

Specific examples of arylcarbonyloxy include phenylcarbonyloxy, naphthylcarbonyloxy, fluorenylcarbonyloxy, anthrylcarbonyloxy, biphenylylcarbonyloxy, tetrahydronaphthylcarbonyloxy, chromanylcarbonyloxy, 2,3-dihydro-1 ,4-dioxanaphthalenylcarbonyloxy, indanylcarbonyloxy and phenanthrylcarbonyloxy.

Specific examples of arylcarbonylamino include phenylcarbonylamino, naphthylcarbonylamino, fluorenylcarbonylamino, anthrylcarbonylamino, biphenylylcarbonylamino, tetrahydronaphthylcarbonylamino, chromanylcarbonylamino, 2,3-dihydro-1 ,4-dioxanaphthalenylcarbonylamino, indanylcarbonylamino and phenanthrylcarbonylamino.

Specific examples of alkoxy include linear or branched C _{1- 6} alkoxy can be mentioned.

Specific examples of aryloxy include phenyloxy, naphthyloxy, fluorenyloxy, anthryloxy, biphenylyloxy, tetrahydronaphthyloxy, chromanyloxy, 2,3-dihydro-1,4-dioxanaphthalenyl. oxy, indanyloxy and phenanthryloxy.

Specific examples of aralkyl include benzyl, naphthylmethyl, fluorenylmethyl, anthrylmethyl, biphenylylmethyl, tetrahydronaphthylmethyl, chromanylmethyl, 2,3-dihydro-1,4-dioxanaphthalenylmethyl, indanylmethyl and phenanthrylmethyl, phenethyl, naphthylethyl, fluorenylethyl, anthrylethyl, biphenylylethyl, tetrahydronaphthylethyl, chromanylethyl, 2,3-dihydro-1,4-dioxanaphthalenyl Ethyl, indanylethyl and phenanthrylethyl are mentioned.

Specific examples of aralkyloxy include benzyloxy, naphthylmethyloxy, fluorenylmethyloxy, anthrylmethyloxy, biphenylylmethyloxy, tetrahydronaphthylmethyloxy, chromanylmethyloxy, 2,3-dihydro-1,4 - dioxanaphthalenylmethyloxy, indanylmethyloxy and phenanthrylmethyloxy, phenethyloxy, naphthylethyloxy, fluorenylethyloxy, anthrylethyloxy, biphenylylethyloxy, tetrahydronaphthylethyloxy, chromanyl Ethyloxy, 2,3-dihydro-1,4-dioxanaphthalenylethyloxy, indanylethyloxy and phenanthrylethyloxy.

Substituents of the optionally substituted benzene ring or optionally substituted pyridine ring include a halogen atom, OH, NO ₂ , CN, alkyl, cycloalkyl, alkoxy, alkylcarbonyloxy, Arylcarbonyloxy, amino, carbamoyl, alkylcarbonylamino, aryl, aralkyl, alkylcarbonyl, SH or alkylthio.

The compounds of the invention can be prepared according to Schemes 1-3 below.
Scheme 1

(Wherein, R ¹ to R ⁴ , R ⁶ , R ⁷ and n are as defined above.)
Scheme 2

(Wherein, R ¹ to R ⁷ , Z and n are as defined above. X represents a leaving group.)
Scheme 3

(Wherein, R ¹ to R ⁷ , Z and n are as defined above. X represents a leaving group.)
In Scheme 1, 1 mol to excess of compound (2), 1 mol to excess of copper sulfate, and 1 mol to excess of ascorbic acid are used relative to 1 mol of compound (1), and click chemistry is performed in the presence of a solvent. The compound (IA) of the present invention wherein R ⁵ =H and Z=N can be obtained by reacting at a temperature of about 10°C to the boiling point of the solvent for 1 to 24 hours.

In scheme 2, compound (4) is used in an excess amount of 1 mol to 1 mol of compound (3), and if necessary, in the presence of a base and a solvent at a temperature of about 10 ° C. to the boiling point of the solvent. The compound of general formula (I) of the present invention can be obtained by reacting for time.

In scheme 3, compound (6) is used in an excess amount of 1 mol to 1 mol of compound (5), and if necessary, in the presence of a base and a solvent at a temperature of about 10 ° C. to the boiling point of the solvent. The compound of general formula (I) of the present invention can be obtained by reacting for time.

In the reactions of Schemes 1 to 3, the solvents include chlorinated hydrocarbons such as methylene chloride and 1,2-dichloroethane, aromatic solvents such as toluene, ester solvents such as ethyl acetate, and ketone solvents such as acetone and methyl ethyl ketone. , diethyl ether, and tetrahydrofuran.

In the reactions of Schemes 2-3, bases include DBU, pyridine, triethylamine, diisopropylethylamine, and the like.

Leaving groups include chlorine atom, bromine atom, p-toluenesulfonyloxy (Ts-O), benzenesulfonyloxy, methanesulfonyloxy (Ms-O) and the like.

Preferred compounds of the present invention are KDM5C (Lysine-specific demethylase 5C) inhibitors, and more preferably selectively inhibit KDM5C. The _IC50 for KDM5C is preferably 100 nM or less, more preferably 50 nM or less, even more preferably 10 nM or less. Also, the _IC50 for KDM5C is preferably at least 10 times lower than the _IC50 for KDM5A, more preferably at least 50 times lower, even more preferably at least 100 times lower.

The compounds of the present invention have potent KDM5C inhibitory activity and are preferred as antidepressants.

The administration method, dosage form, and dosage of a drug containing the compound of the present invention as an active ingredient can be appropriately determined according to the purpose of use. For example, the dosage form of a drug containing the compound of the present invention as an active ingredient may be oral administration or parenteral administration. Dosage forms include oral administration agents such as tablets, powders, capsules, granules, extracts and syrups, and parenteral administration agents such as injections, drip infusions and suppositories. These formulations can be prepared as pharmaceutical compositions containing a compound of the invention and a pharmaceutically acceptable excipient. The effective dose of the compound of the present invention as an antidepressant is usually 0.1-1000 mg per day for adults when administered orally and 0.01-200 mg when administered parenterally. Administer once or multiple times. Since the dose varies depending on various conditions, a dose less than the above dose range may be sufficient.

EXAMPLES The present invention will be described in more detail below with reference to examples, but the present invention is not limited to the following specific examples.

·analysis
Melting points were measured using a Yanagimoto melting point apparatus.
¹ H NMR (300 MHz) and ¹³ C NMR (75 MHz) were measured using a BRUKER AVANCE300 AV spectrometer. Chemical shifts (δ) are shown in parts per million (ppm) based on tetramethylsilane as an internal standard.
Electrospray ionization (ESI) Mass spectra were obtained using a BRUKER HCT plus mass spectrometer.
HPLC was performed by attaching a COSMOSIL Packed Column (5C ₁₈ -AR-II, 4.6ID×150 mm, manufactured by Nacalai) to an HPLC apparatus manufactured by Shimadzu under the following conditions. Solvents: (A) 0.1% TFA aqueous solution, (B) 0.1% TFA acetonitrile solution. Flow rate: 1.0 mL/min. Measurement wavelength: 254 nm. Solvent composition: A/B 0-20 min (90/10 to 10/90), 20-30 min (10/90), 30-40 (10-90 to 90/10).

・Reagents Commercially available reagents and solvents from Aldrich, Tokyo Chemical Industry, Kishida Chemical, Kanto Chemical, Wako Pure Chemical, and Nacalai Tesque were used as they were. For column chromatography, TOYOTA KAKO SILICA GEL (#AP300D) silica gel (particle size: 200-440 mesh) was used.

CPI-455, reported as a KDM5 inhibitor, was synthesized according to the literature (Nat Chem Biol. 2016 Jul;12(7):531-8. doi: 10.1038/nchembio.2085. Epub 2016 May 23.).

NCDM-81a, reported as a KDM5 inhibitor, was synthesized according to the literature (ACS Med. Chem. Lett. 2015, 6, 6, 665-670).

Production example 1
2-Ethynylisoniconic acid (Alk1)
Step 1. methyl 2-bromoisonicotinate (9).
^EDCI.HCl (1.36 g, 7.20 mmol) was added to a dichloromethane/methanol (10.0 mL/15.0 mL) solution of 2-bromonicotinic acid (1.46 g, 7.23 mmol) and stirred at room temperature for 20 hours. After completion of the reaction, the solvent was distilled off and the residue was purified by silica gel column chromatography (ethyl acetate/n-hexane=1/10). 1.35 g of product 9 was obtained as a colorless powdery solid. Yield 87%. ¹ H NMR (CDCl ₃ , 300 MHz, δ; ppm) 8.52 (dd, 1H, J = 4.2, 0.6 Hz), 8.02 (t, 1H, J = 6.6 Hz), 7.80 (dd, 1H, J = 3.9, 0.6 Hz), 3.97 (s, 3H); ¹³ C NMR (CDCl ₃ , 75 MHz, δ; ppm) 151.04, 144.26, 141.19, 129.15, 123.30, 54.40; MS (ESI) m/z 215.7, 217.7 (MH ⁺ ).

Step 2: 2 -trimethylsilylethynylpyridine- 4 -carboxylic acid methyl ester (10)
_{Trimethylsilylacetylene ( _} 165 μL, 1.17 mmol) was added under an argon atmosphere. The reaction solution was stirred at room temperature under an argon atmosphere for 3 hours. After completion of the reaction, the solvent was distilled off, diethyl ether was added to the residue, and the solid was removed by filtration. After concentrating the filtrate, the residue was purified by silica gel column chromatography (ethyl acetate/n-hexane=1/10) to obtain 102 mg of a yellow oily product 10. Yield 70%. ¹ H NMR (CDCl ₃ , 300 MHz, δ; ppm) 8.68 (dd, 1H, J = 4.2, 0.6 Hz), 7.97 (s, 1H), 7.73 (dd, 1H, J = 4.2, 0.6 Hz), 3.93 (s, 3H), 0.26 (s, 9H); ¹³ C NMR(CDCl ₃ , 75 MHz, δ; ppm) 151.04, 144.26, 138.20, 127.06, 122.58, 103.50, 96.86,53.35, 0.18; MS (ESI) m /z 233.8 (MH ⁺ ).

Step 3: 2-Ethynyisonicotinic acid (Alk1)
To a methanol (2.00 mL) solution of compound 10 (50.0 mg, 215 µmol) was added 2 N aqueous sodium hydroxide solution (236 µL, 472 µmol) at 0°C. The reaction solution was stirred at room temperature for 12 hours. After completion of the reaction, a saturated aqueous citric acid solution was added to the reaction solution to adjust the pH to 4.0. The precipitated solid was collected by filtration, washed with water, and dried. The crude product was recrystallized with methanol/ethyl acetate=2/1 to obtain 29.4 mg of white solid Alk1. Yield 93%. mp 203-205 °C; ¹ H NMR (DMSO-d ₆ , 300 MHz, δ; ppm) 8.75 (dd, 1H, J = 5.1, 0.9 Hz), 7.87-7.88 (m, 1H), 7.81 (dd, 1H, J = 4.8, 1.5 Hz), 4.45 (s, 1H); ¹³ C NMR (DMSO-d ₆ , 75 MHz, δ; ppm) 164.95, 150.97, 143.43, 137.88, 126.11, 122.75, 82.64, 81.05; Anal H, _3.43 ; N, 9.52. Found: C _, 64.91 _; H, 3.60; N, 9.28.

N -(methylsulfonyl)-2-ethynyl -4- pyridinecarboxamide (Alk2)Alk1 (147 mg, 1.00 mmol), EDCI.HCl (388 mg, 2.00 mmol), and DMAP (244 mg, 2.00 mmol) in dichloromethane (15.0 mL) ), methyl sulfonamide (189 mg, 2.00 mmol) was added to the solution, and the mixture was stirred at room temperature for 24 hours. The reaction solution was washed with water, and the aqueous layer was acidified with 1N hydrochloric acid and then extracted with dichloromethane. After the organic layer was washed with water and dried with Na ₂ SO ₄ , the solvent was removed by an evaporator. The residue was purified by silica gel column chromatography (dichloromethane/methanol = 10/1) to obtain 56 mg of brown solid Alk2. Yield 26%. mp 141-142°C; ¹ H NMRDMSO-d ₆ , 300 MHz, δ; ppm) 8.76 (1H, dd, J = 4.5, 0.9 Hz), 8.00 (1H, q, J = 0.9 Hz), 7.82 (1H dd, J = 3.6, 1.5 Hz) _, 4.50 (1H, s ⁾ , 3.38 (3H, s); 121.88, 82.41, 81.39, 41.16; _MS ( _ESI ) m/z 225.1 (MH ⁺ ); _HPLC t _R =7.58 min purity 96.5%. Anal. H, 3.60; N, 9.28. HRMS (EI) Calcd. for C8H5O2N 147.0320, Found 147.0310.

4-(2H-tetrazol-5-yl)-pyridine (Alk3)
Step 1:4-cyano-2-[2-(trimethylsilyl)ethynyl]pyridine (12)
2-chloro-4-cyanopyridine 11 (1.40 g, 10.0 mmol), PdCl ₂ (PPh ₃ ) ₂ (138 mg, 330 μmol) and CuI (38.0 mg, 330 μmol) in triethylamine (20.0 mL)/acetonitrile (20.0 mL) ) solution was added with trimethylsilylacetylene (2.20 mL, 15.0 mmol) under an argon atmosphere and the reaction solution was heated at 60 °C for 3 h under an argon atmosphere. After completion of the reaction, the solvent was distilled off with an evaporator, diethyl ether was added, and the organic layer was washed with 1N hydrochloric acid, saturated NaHCO ₃ aqueous solution and brine, and dried with Na ₂ SO ₄ . After distilling off the solvent, the residue was purified by column chromatography (ethyl acetate/n-hexane = 1/10) to obtain 1.96 g of a yellowish white solid 12. Yield 98%. mp 46.0-48.0 °C ¹ H NMR (CDCl ₃ , 300 MHz, δ; ppm) 8.75 (1H, dd, J = 4.2, 0.9 Hz), 7.67 (1H, q, J = 0.3 Hz), 7.44 (1H, ^dd , J = 3.6, 1.5 Hz) _, 0.30 (9H, s); (ESI) m/z 201.1 (MH ⁺ ).

Step 2: 4-cyano-2-ethynylpyridine (13)
To a solution of compound 12 (1.96 g, 9.79 mmol) in dichloromethane (25.0 mL) was added 1 M TBAF/THF (14.0 mL, 14.0 mmol) at 0°C, and the reaction solution was stirred at 0°C for 2 hours. After completion of _the reaction, ethyl acetate was added and the organic layer was washed with water and dried with _Na2SO4 . After distilling off the solvent, the residue was purified by silica gel column chromatography (ethyl acetate/n-hexane=1/3) to obtain 823 mg of a yellowish white solid 13. Yield 66%. mp 104-106 °C; ¹ H NMR (CDCl ₃ , 300 MHz, δ; ppm) 8.76 (1H, dd, J = 4.2, 0.9 Hz), 7.68 (1H, q, J = 0.3 Hz), 7.49 (1H ¹³ C NMR (CDCl ₃ , 75 MHz, δ; ppm) 151.11, 143.79, 128.84, 124.70, 121.08, 115.70, 81.10, 80.05; MS ( ESI) m/z 129.2 (MH ⁺ ).

Step 3: 4-(2H-tetrazol-5-yl) -2-ethynyl- pyridine (Alk3)
Sodium azide (195 mg, 3.00 mmol) and ammonium chloride (162 mg, 3.00 mmol) were added to a solution of compound 13 (396 mg, 3.00 mmol) in DMF (20.0 mL) under nitrogen atmosphere. The reaction solution was heated and stirred at 80° C. for 12 hours under a nitrogen atmosphere. After completion of the reaction, ethyl acetate was added and washed with 4N hydrochloric acid. _The organic layer was washed with water, brine and dried over _Na2SO4 . After distilling off the solvent, the residue was purified by silica gel column chromatography (ethyl acetate/n-hexane=2/1) to obtain 62.0 mg of brown solid Alk3. Yield 12%. mp 210-212°C; ¹ H NMR (DMSO-d ₆ , 300 MHz, δ; ppm) 8.79(1H, dd, J = 4.2, 0.9 Hz), 8.10 (1H, q, J = 0.6 Hz), 8.00 ( 1 H, dd, J = _3.3 ^, 1.8 Hz), 4.51 (1 H, s); ; MS (ESI) m/z 172.1 (MH ⁺ ); HPLC t _R =7.45 minpurity 96.0%. HRMS (EI) Calcd. for C8H5N5 171.0537, Found 171.0545.

Methyl- 2-ethynylisonicotinate (Alk4)
To a solution of compound 10 (380 mg, 1.60 mmol) in THF (2.00 mL) was added 1 M TBAF/THF (2.20 mL, 2.20 mmol) at 0°C, and the reaction solution was stirred at 0°C for 2 hours. After completion of _the reaction, ethyl acetate was added, washed with water, and the organic layer was dried with _Na2SO4 . After distilling off the solvent, the residue was purified by silica gel column chromatography (ethyl acetate/n-hexane=1/2). Furthermore, 248 mg of colorless solid Alk4 was obtained by recrystallizing the crude product with n-hexane. Yield 96%. mp 64.5-65.5°C; ¹ H NMR (CDCl ₃ , 300 MHz, δ; ppm) 8.75 (d, 1H, J = 5.1 Hz), 8.03 (q, 1H, J = 0.6 Hz), 7.81 (dd, 1H, J = 4.2, 0.9 Hz), 3.97 (s ^, 3H) _, 3.23 (s, 1H); 78.37, 53.00 ; MS (ESI) m/z 162.2 (MH ⁺ ); HPLC t _R = 11.7 min purity 99.4%

Production example 2
Synthesis of Azide Compounds Az1-7, YMA04-110-A, YMA04-110-D
Step 1: 2-Azidoethanol (15)
Sodium azide (647 mg, 8.30 mmol) was added to an aqueous solution (5.00 mL) of 2-bromoethanol (581 μL, 10.0 mmol), and the reaction solution was heated to reflux at 100° C. for 12 hours. After completion of the reaction, sodium chloride was added and the aqueous layer was extracted with dichloromethane. The organic layer was dried over Na ₂ SO ₄ and the solvent was distilled off. The resulting colorless liquid crude product was used directly in the next reaction.

Step 2: 1-Mesyl-2-azidoethanol (16)
Triethylamine (1.74 mL, 12.5 mmol) and methanesulfonyl chloride (967 μL, 12.5 mmol) were added to a solution of compound 15 in dichloromethane (10.0 mL) at 0°C, and the reaction solution was stirred at room temperature for 12 hours. After completion of the reaction, the reaction solution was filtered, and the filtrate was concentrated by an evaporator. The residue was purified by silica gel column chromatography (ethyl acetate/n-hexane=2/3) to obtain 560 mg of 16 as a colorless oil. Yield 34%. ¹ H NMR (CDCl ₃ , 300 MHz, δ; ppm) 4.26 (2H, t, J = 5.7 Hz), 3.52 (2H, t, J = 4.5 Hz), 3.01 (3H, s); ¹³ C NMR (CDCl ₃ , 75 MHz, δ; ppm) 67.88, 49.55, 37.17.

Step 3: 2-Azido-1-ethylhexylmethylamine (Az1)
A solution of compound 16 (189 mg, 1.15 mmol) and potassium carbonate (482 mg, 3.50 mmol) in acetonitrile (5 mL) was added with N-hexylmethylamine (151 μL, 1.00 mmol) and heated at 90 °C for 12 h. refluxed. After completion of the reaction, the reaction solution was filtered, and the filtrate was concentrated by an evaporator. The residue was purified by silica gel column chromatography (ethyl acetate/n-hexane=1/3) to obtain 50.0 mg of Az1 as a colorless oil. Yield 27%. ¹ H NMR (CDCl ₃ , 300 MHz, δ; ppm) 3.32 (2H, t, J = 6.0 Hz), 2.58 (2H, t, J = 6.0 Hz), 2.37 (2H, t, J = 7.8 Hz), 2.26 (3H, s), 1.42-1.49 (2H, m), 1.26-1.32 (6H, m), 0.89 (3H, t J= 6.6 Hz); ¹³ C NMR (CDCl ₃ , 75 MHz, δ; ppm) 58.14, 56.52, 49.16, 42.34, 31.91, 27.36, 27.16, 22.74, 14.14; MS (ESI) m/z 185.1 (MH ⁺ ); HPLC t _R =10.2 min purity 97.6%. , Found 185.1761.

Azides Az2-7, YMA04-110-A and YMA04-110-D were synthesized by the same method as shown in the synthetic method of Az1.
2-Azido-1-benzylmethylethylamine (Az2)
Yield 53%; colorless oil ¹ H NMR (CDCl ₃ , 300 MHz, δ; ppm): 7.28-7.40 (5H, m), 3.61 (2H, s), 3.36 (2H, t, J = 6.0 Hz), 2.69 (2H, t, J = 6.0 Hz), 2.31 (3H, s); ¹³ C NMR (CDCl ₃ , 75 MHz, δ; ppm): 138.68, 128.91, 128.34, 127.15, 62.57, 56.26, 48.90, 42.15; MS (ESI) m/z 191.2 (MH ⁺ ). HPLC t _R =7.87 min purity 96.2%.

2-Azido-1-diethylbutylamine (Az3)
Yield 24%; colorless oil ¹ H NMR (CDCl ₃ , 300 MHz, δ; ppm): 3.28 (2H, t, J = 6.3 Hz), 2.66 (2H, t, J = 6.3 Hz), 2.55 (2H, q, J = 6.9 Hz), 2.46 (2H, t, J = 7.8 Hz), 1.31-1.44 (4H, m), 1.04 (3H, t, J = 0.9 Hz), 0.93 (3H, t, J = 7.2 Hz); ¹³ C NMR (CDCl ₃ , 75 MHz, δ; ppm): 53.60, 52.85, 49.51, 47.81, 29.37, 20.55, 14.02, 11.80; MS (ESI) m/z 171.3 (MH ⁺ ) _. =22.9 min purity 95.8%.

2-Azido-1-ethylpentylmethylamine (Az4)
Yield 37%; colorless oil ¹ H NMR (CDCl ₃ , 300 MHz, δ; ppm): 3.32 (2H, t, J= 6.3 Hz), 2.57 (2H, t, J = 6.0 Hz), 2.36 (2H, t, J = 6.0 Hz), 2.26 (3H, s), 1.42-1.52 (2H, m), 1.28-1.32 (4H, m), 0.90 (3H, t, J = 7.2 Hz); ¹³ C NMR (CDCl ₃ , 75 MHz, δ; ppm): 57.95, 56.38, 49.00, 42.18, 29.52, 26.92, 22.57, 14.00; MS (ESI) m/z 171.3 (MH ⁺ ). HPLC t _R = 22.7 min purity 98.5%.

2-Azido-1-ethylpropylbutylamine (Az5)
Yield 36%; colorless oil ¹ H NMR (CDCl ₃ , 300 MHz, δ; ppm): 3.25 (2H, t, J = 6.0 Hz), 2.65 (2H, t, J = 6.3 Hz), 2.39-2.47 ( 4H, m), 1.30-1.50 (6H, m), 0.89 (6H, q, J = 7.2 Hz); ¹³ C NMR (CDCl ₃ , 75 MHz, δ; ppm): 56.49, 54.20, 53.56, 49.54, 29.38 , 20.52, 20.38, 14.01, 11.76; MS (ESI) m/z 185.3 (MH ⁺ ). HPLC t _R = 10.2 min purity 97.6%

2-Azido-1-ethyldibutylamine (Az6)
Yield 48%; colorless oil ¹ H NMR (CDCl ₃ , 300 MHz, δ; ppm): 3.25 (2H, t, J = 6.3 Hz), 2.63 (2H, t, J = 6.3 Hz), 2.44 (4H, t, J = 7.2 Hz), 1.30-1.43 (8H, m), 0.92 (6H, t, J = 7.2 Hz); ¹³ C NMR (CDCl ₃ , 75 MHz, δ; ppm): 54.19, 53.52, 49.53, 29.39, 20.51, 13.99; MS (ESI) m/z 199.3 (MH ⁺ ). HPLC t _R = 9.15 min purity 99.9%.

2-Azido-1-ethylbutylbenzylamine (Az7)
Yield 16%; colorless oil ¹ H NMR (CDCl ₃ , 300 MHz, δ; ppm): 7.25-7.39 (5H, m), 3.65 (2H, s), 3.26 (2H, t, J = 6.0 Hz), 2.71 (2H, t, J = 6.3 Hz), 2.51 (2H, t, J = 7.2 Hz), 1.47-1.57 (2H, m), 1.29-1.41 (2H, m), 0.92 (3H, t, J = 7.2 Hz); ¹³ C NMR (CDCl ₃ , 75 MHz, δ; ppm): 139.49, 128.73, 128.23, 126.95, 59.09, 54.13, 53.31, 49.43, 29.29, 20.44, 14.01; MS (ESI) m/z 233.3 ( MH ⁺ ). HPLC _tR = 11.1 min purity 96.3%

1-(2-Azidoethyl)piperidine (YMA04-110-A)
Yield 42%; yellow oil ¹ H NMR (CDCl ₃ , 300 MHz, δ; ppm) 3.31 (2H, t, J = 6.3 Hz), 2.52 (2H, t, J = 6.3 Hz), 2.41 (4H, t , J = 5.4 Hz), 1.53-1.59 (4H, m ⁾ , 1.37-1.45 ₍ 2H, m); ; MS (ESI) m/z 155.0 (MH ⁺ )

2-Azido-N-methyl-N-(phenyl methyl)ethanamine (YMA04-110-D)
Yield 43%; yellow oil: ¹ H NMR (CDCl ₃ , 300 MHz, δ; ppm) 7.28-7.40 (5H, m), 3.61 (2H, s), 3.36 (2H, t, J = 6.0 Hz), 2.69 (2H, t, J = 6.0 Hz), 2.31 (3H, s); ¹³ C NMR (CDCl ₃ , 75 MHz, δ; ppm) 138.7, 128.9, 128.3, 127.2, 62.57, 56.26, 48.90, 42.15; (ESI) m/z 190.1 (MH ⁺ )

Production example 3
2-Azido-1-ethylbutylhexylamine (Az8)
Step 1: N-butyl-1-hexanamine (17)
BH ₃ ^.SMe ₂ (1.00 mL, 10.0 mmol) was added dropwise to 1-hexene (5.00 mL, 40.0 mmol) at 0° C. under nitrogen atmosphere. After the dropwise addition was completed, the reaction solution was stirred at room temperature for 12 hours under a nitrogen atmosphere. It was added dropwise under a nitrogen atmosphere. This solution was added quickly to the trialkylborane solution at 0°C under a nitrogen atmosphere. The reaction solution was stirred at room temperature for 3 hours under a nitrogen atmosphere. After completion of the reaction, 10% HCl was added to adjust the reaction solution to pH 1.0 and washed with diethyl ether. A 6 N NaOH aqueous solution was added to the aqueous layer to adjust the pH to >13, and the desired product was extracted with ether. The organic layer was dried over Na ₂ SO ₄ and concentrated by an evaporator to obtain 650 mg of product 17 as a colorless oil. Yield 41%. ¹ H NMR (CDCl ₃ , 300 MHz, δ; ppm) 2.56 (2H, t, J = 7.2 Hz), 2.56 (1H, t, J = 7.5 Hz), 1.26-1.47 (12H, m), 0.83-0.99 (6H, m); ^13C NMR ( _CDCl3 , 75 MHz, δ; ppm) 50.16, 49.81, 32.32, 31.79, 30.15, 27.09, 22.60, 20.52, 14.00; MS (ESI) m/z 158.3 (MH ⁺ ).

Step 2: 2-Azido-1-ethylbutylhexylamine (Az8)
Compound 17 (157 mg, 1.00 mmol) was added to a solution of compound 16 (189 mg, 1.15 mmol) and potassium carbonate (482 mg, 3.50 mmol) in acetonitrile (5.00 mL), and the mixture was heated under reflux at 90 °C for 12 hours. After completion of the reaction, the reaction solution was filtered, and the filtrate was concentrated by an evaporator. The residue was purified by silica gel column chromatography (ethyl acetate/n-hexane=1/3) to obtain 104 mg of Az8 as a colorless oil. Yield 46%. ¹ H NMR (CDCl ₃ , 300 MHz, δ; ppm): 3.26 (2H, t, J = 6.0 Hz), 2.64 (2H, t, J = 6.3 Hz), 2.44 (2H, t, J = 7.2 Hz) , 2.44 (2H, t, J = 7.2 Hz), 1.40-1.45 (4H, m), 1.27-1.32 (8H, m), 0.87-0.94 (6H, m); ¹³ C NMR (CDCl ₃ , 75 MHz, δ; ppm): 54.67, 54.35, 53.64, 49.68, 31.96, 29.50, 27.29, 27.24, 22.80, 20.70, 14.19; MS (ESI) m/z 227.3 (MH ⁺ ). HPLC t _R = 13.3 min purity 98.3%

4-(2-Azidoethyl)morpholine (YMA04-112)
Bromoethanol (349 μL, 5.00 mmol) was added dropwise to a toluene solution (20 mL) of morpholine (948 μL, 10.0 mmol), and the reaction solution was heated under reflux for 3 hours. After completion of the reaction, the reaction solution was filtered, and the filtrate was concentrated by an evaporator. The residue was dissolved in dichloromethane (8 mL) and trimethylamine (1.67 mL, 12.0 mmol) was added. After methanesulfonyl chloride (902 μL, 12.0 mmol) was added dropwise at 0 ^° C., the reaction solution was stirred at room temperature for 3 hours. After completion of the reaction, saturated aqueous sodium bicarbonate was added and the mixture was extracted with dichloromethane. The organic layer was washed with brine, dried over Na ₂ SO ₄ and the solvent was removed by an evaporator. DMSO (30 mL) and sodium azide (780 mg, 12.0 mmol) were added to the residue, and the reaction solution was heated with stirring at 80 ^° C. for 2 hours. After completion of the reaction, water was added and the mixture was extracted with ethyl acetate. The organic layer was washed with water, dried over Na ₂ SO ₄ , and the solvent was removed by an evaporator. The residue was purified by silica gel column chromatography (ethyl acetate) to obtain yellow oily 4-(2-Azidoethyl)morpholine (YMA04-112, yield 18%). ¹ H NMR (CDCl ₃ , 300 MHz, δ; ppm) 3.69-3.78 (4H, m), 3.34 (2H, t, J = 6.0 Hz), 2.57 (2H, t, J = 1.5 Hz), 2.49-2.50 (4H, m); ¹³ C NMR (CDCl ₃ , 75 MHz, δ; ppm) 66.95, 57.67, 53.68, 48.01; MS (ESI) m/z 157.1 (MH ⁺ )

Synthesis Example 1 of Triazole Compounds NPC-3422, YMA04-115, YMA04-118, YMA04-119
2 -(1-( 2-( hexyl ( methyl ) amino o)ethyl) -1H-[1,2,3]triazol- 4 -yl)-pyridine- 4 -carboxylic acid (NPC-3422)
Step 1: Methyl-2 -(1-( 2-( hexyl ( methyl ) amino o)ethyl) -1H-[1,2,3]triazol- 4 -yl)-pyridine- 4 -carboxylate (NPC-3543)
CuSO4 5H2 O (25.0 mg, 100 μmol) and sodium ascorbate (20.0 mg, 100 μmol) was added and the reaction solution was stirred at room temperature for 12 hours. After completion of the reaction, the solvent was distilled off using an evaporator, water was added, and the mixture was extracted with ethyl acetate. _The organic layer was washed with brine and dried over _Na2SO4 . After distilling off the solvent, the residue was purified by silica gel column chromatography (ethyl acetate/n-hexane=1/3 to dichloromethane/methanol=5/1) to obtain 16.2 mg of orange oil NPC-3543. Yield 47%. ¹ H NMR (CDCl ₃ , 300 MHz, δ; ppm) 8.71-8.74 (2H, m), 8.30 (1H, s), 7.78 (1H, dd, J = 3.6, 0.6 Hz), 4.52 (2H, t, J = 6.3 Hz), 3.99 (3H,s), 2.88 (2H, t, J = 6.0 Hz), 2.40 (2H, t, J = 7.5 Hz), 2.31 (3H, s), 1.22-1.42(8H, m), 0.83 (3H, t, J = 6.6 Hz); ¹³ C NMR (CDCl ₃ , 75 MHz, δ; ppm) 165.60, 151.70, 150.21, 147.54, 138.35, 123.25, 121.74, 119.50, 57.98, 52.03, 57.03 , 48.57,42.09, 31.75, 27.21, 27.02, 22.63, 14.02; MS (ESI) m/z 346.4 (MH ⁺ ); HPLC t _R = 12.13 min purity 95.6%

Step 2: 2- (1-( 2-( hexyl ( methyl ) amino )ethyl) -1H-[1,2,3]triazol- 4 -yl)-pyridine- 4 -carboxylic acid hydrochloride (NPC-3422 HCl salt )
NPC-3543 (16.2 mg, 47.0 μmol) was dissolved in methanol (3.00 mL), and 2.00 mL of 2 N aqueous sodium hydroxide solution was added. The reaction solution was stirred at room temperature for 12 hours. After completion of the reaction, the solvent was distilled off with an evaporator, and 4N HCl/dioxane was added to the residue to adjust the pH to 1.0. The solution was filtered and the filtrate was concentrated to give 10.0 mg of colorless solid NPC-3422 (HCl salt). Yield 25% ¹ H NMR (MeOD, 300 MHz, δ; ppm) 8.79 (1H, d, J = 4.5 Hz), 8.64 (1H, s), 8.61 (1H, s), 7.89 (1H,dd, J = 3.6, 1.5Hz), 4.99 (2H, t, J = 6.0Hz), 3.92 (2H, s), 3.30 (2H, s), 3.02(3H, s), 1.73 (2H, quin., J = 7.8 Hz), 1.38 (6H, m), 0.93 ( ^3H , t, J = 7.2 Hz); , 56.67, 54.24, 44.53, 39.71, 30.92, 25.73, 23.56, 22.03, 12.79; MS(ESI) m/z 332.1(MH ⁺ ); HPLC t _R = 10.10 min purity 99.6% HRMS (EI) Calcd. , Found 331.2041.

Triazole compounds YMA04-115, YMA04-118 and YMA04-119 were synthesized using the corresponding azide compounds as starting materials in the same manner as for the synthesis of NPC-3422.

2 -(1-( 2-( methyl ( benzyl ) amino o)ethyl) -1H-[1,2,3]triazol- 4 -yl)-pyridine- 4 -carboxylic acid hydrochloride (YMA04-115)
Yield 66%; yellowish white solid ¹ H NMR (DMSO-d ₆ , 300 MHz, δ; ppm): 8.82 (2H, s), 8.45 (1H,s), 7.78 (1H, d, J = 3.9 Hz) , 7.59-7.56 (2H, m), 7.48-7.46 (3H, m), 5.01 (2H, t, J = 6.6 Hz), 4.40 (2H, s), 3.72 (2H, t, J = 3.0 Hz), 2.73 (3H, s); ^13C NMR (DMSO- _d6 , 75 MHz, δ; ppm): 166.5, 159.1, 151.4, 147.4, 139.8, 131.9, 130.3, 130.2,129.4, 125.1, 122.5, 118.8, 59.5 53.9, 45.0; HRMS (EI) Calcd. for C18H19O2N5 (MH ⁺ ) 338.1612, Found 338.1609(MH ⁺ ). HPLC t _R =11.87 min purity 96.2%.

2 -( 2 -( 1- piperid yl)ethyl) -1H-[1,2,3]triazol- 4 -yl)-pyridine- 4 -carboxylic acid hydrochloride (YMA04-118 HCl salt )
Yield 38%; colorless solid ¹ H NMR (DMSO-d ₆ , 300 MHz, δ; ppm): 8.88 (1H, s), 8.80 (1H, d, J = 4.8 Hz), 8.44 (1H, s), 7.77 (1H, d, J = 3.9 Hz), 5.02 (2H, t, J = 6.6 Hz), 3.67 (2H, d, J = 4.2 Hz), 3.41 (2H, d, J = 3.9 Hz), 2.99- 2.93 (2H, m), 1.84-1.35 (6H, m); ¹³ C NMR (DMSO-d ₆ , 75 MHz, δ; ppm): 166.3, 151.1, 151.0, 147.1, 139.9, 124.9, 122.4, 118.8, 54.5 , 52.6, 44.6, 22.6, 21.6; MS (ESI) (MH ⁺ ). HRMS (EI) Calcd. for C15H19O2N5 (MH ⁺ ) 302.1612, _Found 302.1610(MH ⁺ ).

2 -( 2 -( 4- morpholin yl)ethyl) -1H-[1,2,3]triazol- 4 -yl)-pyridine- 4 -carboxylic acid hydrochloride (YMA04-119 HCl salt )
Yield 35%; orange solid ¹ H NMR (DMSO-d ₆ , 300 MHz, δ; ppm): 8.82 (2H, s), 8.45(1H, s), 7.78 (1H, d, J = 3.9 Hz) , 7.59-7.56 (2H, m), 7.48-7.46 (3H, m), 5.01 (2H, t, J = 6.6 Hz), 4.40 (2H, s), 3.72 (2H, t, J = 3.0 Hz), 2.73 (3H, s); ¹³ C NMR (DMSO-d ₆ , 75 MHz, δ; ppm): 166.5, 159.1, 151.4, 147.4, 139.8, 131.9, 130.3, 130.2, 129.4, 125.1, 122.5, 118.5, 59 53.9, 45.0; HRMS (EI) Calcd. for C14H17O3N5 (MH ⁺ ) 304.1404, Found 304.1406 (MH ⁺ ). HPLC t _R =1.75 min purity 96.0%.

2- (1-( 2-( hexyl ( methyl ) amino )ethyl) -1H-[1,2,3]triazol-5-yl)-pyridine-4-carboxylic acid (syn-T1 ) and NPC-342 2
Step 1: Methyl-2 -(1-( 2-( hexyl ( methyl ) amino )ethyl) -1H-[1,2,3]triazol-5-yl)-pyridine-4-carboxylate (syn-T4 ) and NPC-354 3
Alk4 (123 mg, 0.760 mmol) and Az1 (210 mg, 1.14 mmol) were dissolved in 5.00 mL of toluene and heated under reflux at 100° C. for 30 hours. After completion of the reaction, the solvent was evaporated, and the residue was purified by silica gel column chromatography (ethyl acetate/n-hexane = 1/1 to 1/0) to give 76.0 mg of yellow oily syn-triazole (yield 29%). ) (syn-T4) and 68.0 mg of orange oily anti-triazole (26% yield) (NPC-3543), respectively.
syn-T4: ¹ H NMR (CDCl ₃ , 300 MHz, δ; ppm) 8.79 (1H, dd, J = 4.2, 0.9 Hz), 8.12 (1H, t, J = 0.9 Hz), 8.02 (1H, s) , 7.80 (1H, dd, J = 3.6, 1.5 Hz), 4.97 (2H, t, J = 7.2 Hz), 3.96 (3H, s), 2.83 (2H, t, J = 7.2 Hz), 2.32 (2H, t, J = 7.2 Hz), 2.22(3H, s), 1.15-1.29 (8H, m), 0.82 (3H, t, J = 7.2 Hz); ¹³ C NMR (CDCl ₃ , 75 MHz, δ; ppm) 164.94, 150.46, 148.46, 148.46, 138.72, 135.70, 133.77, 122.01, 122.01, 57.75, 53.02, 53.02, 47.80, 42.34, 31.76, 31.76, ^27.98 , 22.98 _tR = 12.65 min purity 95.6%
NPC-3543: ¹ H NMR (CDCl ₃ , 300 MHz, δ; ppm) 8.71-8.74 (2H, m), 8.30 (1H, s), 7.78(1H, dd, J= 3.6, 0.6 Hz), 4.52 ( 2H, t, J = 6.3Hz), 3.99 (3H, s), 2.88 (2H, t, J = 6.0 Hz), 2.40 (2H, t, J = 7.5 Hz), 2.31 (3H, s), 1.22- 1.42 (8H, m), 0.83 (3H, t, J= 6.6 Hz); ¹³ C NMR (CDCl ₃ , 75 MHz, δ; ppm) 165.60, 151.70, 150.21, 147.54,138.35, 123.25, 121.74, 119.50, 57.98 , 57.03, 52.70, 48.57, 42.09, 31.75, 27.21, 27.02, 22.63, 14.02; MS (ESI) m/z 346.4 (MH ⁺ ); HPLC t _R = 12.13 min purity 95.6%

Step 2: 2 -(1-( 2-( hexyl ( methyl ) amino )ethyl) -1H-[1,2,3]triazol-5-yl)-pyridine-4-carboxylic acid (syn-T1 ) and NPC-342 2
To a methanol solution (3.00 mL) of syn-isomer and anti-triazole compound (50.0 mg, 145 μmol), 2.00 mL of 2 N aqueous sodium hydroxide solution was added, and the reaction solution was stirred at room temperature for 12 hours. After completion of the reaction, the solvent was distilled off, and 4N hydrochloric acid/dioxane was added to the residue to adjust the pH to 1.0. The solution was filtered and the filtrate was concentrated to give 13.9 mg of syn-T1 and 10.0 mg of anti-T1. (29%/21% yield).
syn-T1: ¹ H NMR (MeOD, 300 MHz, δ; ppm) 8.92 (1H, dd, J = 4.5, 0.6 Hz), 8.43 (1H, s), 8.40 (1H, t, J = 0.3 Hz), 7.97 (1H, dd, J = 3.6, 1.5 Hz), 5.36 (2H, t, J = 6.0 Hz), 3.92 (2H, s), 3.29 (2H, t, J = 8.4 Hz), 3.05 (3H, s ), 1.75 (2H, quin., J = 8.1 Hz), 1.37 (6H, m), 0.93 (3H, t, J = 7.2 Hz); ¹³ C NMR (MeOD, 75 MHz, δ; ppm) 166.99, 151.80 , 148.47, 141.77, 137.53, 135.20 ^, 124.35, 123.72, 58.00, 56.08, 46.15, 41.34, 32.32, 27.14, 25.03, 23.41, _14.21 ; Purity 99.5%
NPC-3422: ¹ H NMR (MeOD, 300 MHz, δ; ppm) 8.79 (1H, d, J = 4.5 Hz), 8.64 (1H, s), 8.61 (1H, s), 7.89 (1H, dd, J = 3.6, 1.5 Hz), 4.99 (2H, t, J = 6.0 Hz), 3.92 (2H, s), 3.30 (2H, s), 3.02 (3H, s), 1.73 (2H, quin., J = 7.8 Hz), 1.38 (6H, m), 0.93 ( ^3H , t, J = 7.2 Hz); , 56.67, 54.24, 44.53, 39.71, 30.92, 25.73, 23.56, 22.03, 12.79; MS (ESI) m/z 332.1(MH ⁺ ); HPLC t _R = 10.10 min purity 99.6%

Test example 1
Enzyme activity test
Alpha screen assay system was used to evaluate the activity of KDM enzyme. The buffers and reagents used for evaluation are shown in Table S1. 2.5 μL of inhibitor solution (3.0% DMSO/assay buffer solution) or assay buffer (50 mM HEPES pH 7.5, 0.1% w/v BSA, 0.01% v/v Tween-20 containing 3% DMSO, for control and blank) was added to each well of the OptiPlate ^™ -384 white plate. 5.0 μL of enzyme solution was added there. (5.0 μL assay buffer was added to blank wells). After 5 minutes at room temperature, 2.5 μL of substrate protein/2-OG (50 μM)/Fe(II) (5 μM)/sodium ascorbate (100 μM) buffer solution was added to each well. The final concentrations of enzymes and substrate proteins are summarized in Table S2. These reaction solutions were stirred at room temperature for 1 or 2 hours with gentle shaking (250 rpm). 5.0 μL of an epigenetic buffer solution of acceptor beads (100 μg/mL) was added to each well and gently shaken (250 rpm) at room temperature for 1 hour. Finally, 10 μL of an epigenetic buffer solution (50 μg/mL) of donor beads was added and shaken at room temperature for 30 minutes in the dark (250 rpm). Alpha signals were then detected with Ensight ^(R) readers (PerkinElmer Ltd). (Excitation wavelength: 615 nm, emission wavelength: 655 nm). Enzyme activity (%) was calculated by dividing the Alpha signal of inhibitor wells by the signal of control wells. The results are shown in FIG.

Cell-cultured human prostate cancer cell PC3 (Japanese Collection of Research Bioresources; JCRB9110, Japan) was mixed with 10% fetal bovine serum albumin (FBS; SIGMA, #172012-500ML), 5% penicillin/streptomyc (Nacalai, #09366-44) was cultured in Ham's F-12K (Kaighn's) medium (Gibco, #21127022) containing 5% CO ₂ at 37°C. Human prostate cancer cells LNCaP (American type culture collection, ATCC) and human gastric cancer cells MKN45 (provided by RIKEN BRC cell bank; RCB1001, Japan) were incubated with RPMI-1640 (Sigma, R8758) medium, cultured at 37°C under 5% CO ₂ atmosphere.

western blotting
PC3, LnCAP and MKN45 cells (5 x ¹⁰⁵ cells/2 mL/dish) were treated with NPC-3543 and cultured for 48 hours. Cell pellets were then extracted with SDS buffer. The protein concentration of the extracted lysate was determined using the BCA protein assay. After adjusting the protein concentration of the lysate, electrophoresis was performed using a 5-20% SDS-polyacrylamide gel. Further transferred to PVDF membrane. After blocking the membrane with 5% skim milk TBS-T solution, primary antibodies polyclonal H3K4me3 antibody (Abcam, #ab8580) (1:5000 dilution), H3K4me2 antibody (CST, #9725) (1:5000 dilution), H3K4me1 antibody (Abcam, #ab8895) (1:5000 dilution), H3 antibody (Abcam, #ab1791) (1:200000 dilution), KDM5A antibody (CST, #3876) (1:1000 dilution), KDM5B antibody (CST, # 3273) (1:1000 dilution), KDM5C antibody (CST, #5361) (1:1000 dilution), mouse monoclonal α-tubulin antibody (sigma, #T8203) (1:1000 dilution) each in 5% skim milk TBS-T solution and reacted with shaking and stirring at room temperature for 1 hour. After washing the membrane three times with TBS-T, the secondary antibody ECL rabbit/mouse IgG, HRP-linked whole Anti-body (GE Healthcare Life Sciences, #NA934) (diluted 1:2500) was reacted at room temperature for 1 hour. Washed again with TBS-T three times. Bands were detected by chemiluminescence using Immobilon ^™ Western Chemiluminescent HRP Substrate (Millipore, #P90718) as detection reagent. The result is shown in figure 2. H3K4 was hypermethylated only in KDM5C-overexpressing PC3 cells. This reveals that NPC-3543 also selectively inhibits KDM5C in cell lines.

Test example 2
Behavioral evaluation test of NPC-3543-administered mice (Figure 3-6)
A cannula was placed in the medial prefrontal cortex of 8-week-old male DBA/2 mice (hereinafter referred to as DBA mice), NPC-3543 (100 μM) or vehicle was administered, and 2 hours later, social defeat stress (*1) was applied. loaded. After doing this for 5 consecutive days, a social test (*2) was conducted.

*1 Social defeat stress test mice are placed in a cage of highly aggressive CD-1 mice for 5 min. At this time, the CD-1 mice unilaterally attack the test mice (physical stress load). After 5 minutes have passed, the CD-1 mouse and the test mouse are separated by a transparent acrylic plate and kept for 24 hours (psychological stress load). This was done for 5 consecutive days (Fig. 3).

*2Sociability test
A CD-1 mouse is placed as a novel mouse in a 42 cm square box (target area). A test mouse (DBA mouse) was placed in this box, and the time spent in the target area for 3 minutes was measured by a video tracking system (Any-Maze software) (Fig. 4).

<Method of sucrose preference test>
A 1% sucrose solution and a bottle of normal water were given simultaneously. The amount of sucrose solution and normal water ingested in 4 hours was measured, and the sucrose preference was used as an index of anhedonia (Fig. 5). Animals usually prefer sweet sucrose, but animals in anhedonia, one of the conditions of depression, become less selective for sucrose.

A KDM5C inhibitor (NPC-3543) showed antidepressant effects in the Sucrose Preference trial. From this, it was shown that the KDM5C inhibitor (NPC-3543) is effective as an antidepressant.

<Gene expression analysis (Fig. 6)>
Stress-vulnerable DBA mice were given mild stress for 5 days. At this time, NPC-3543 (100 μM) or solvent (vehicle, water) was administered daily, gene expression analysis (RNA-seq) using a next-generation sequencer was performed, and principal component analysis (PCA) and clustering analysis were performed. . As a result, it was revealed that the abnormal gene expression pattern caused by stress load was normalized by KDM5C inhibitor (NPC-3543).

Claims (4)

Formula (I) below

(wherein any one of R ¹ to R ⁴ represents COR ⁸ , and the other three of R ¹ to R ⁴ are the same or different, hydrogen atom, halogen atom, OH, NO ₂ , CN, alkyl, cycloalkyl, alkoxy, alkylcarbonylamino, alkylcarbonyloxy, arylcarbonyloxy, carbamoyl, arylcarbonylamino, aryl, aralkyl, alkylcarbonyl, SH or alkylthio R ¹ and R ² , R ² and R ³ , R ³ and R ⁴ represent methylenedioxy, or together with the carbon atoms to which they are attached, a cyclopentene ring, cyclopentadiene ring, cyclohexene ring, cyclohexadiene ring, benzene ring, or a 5-membered or A 6-membered heterocycle may be formed.
^R5 represents a hydrogen atom, a halogen atom, alkyl, alkoxy, aryl, aralkyl, hydroxyalkyl or cycloalkyl. R ⁴ and R ⁵ together with the carbon atom to which they are bonded may form an optionally substituted benzene ring or an optionally substituted pyridine ring.
R ⁶ and R ⁷ are the same or different and represent a hydrogen atom, alkyl, alkoxy, aryl, aralkyl, hydroxyalkyl or cycloalkyl. However, ^R6 and ^R7 are not hydrogen atoms at the same time.
R ⁸ represents OH, alkoxy, hydroxyalkyloxy, cycloalkyloxy, aryloxy or aralkyloxy.
Z indicates N.
n represents an integer of 0-5. )
A compound represented by or a pharmaceutically acceptable salt or solvate thereof. A pharmaceutical composition comprising a compound of claim 1 or a pharmaceutically acceptable salt or solvate thereof and a pharmaceutically acceptable excipient. A KDM5C inhibitor comprising the compound according to claim 1 or a pharmaceutically acceptable salt or solvate thereof as an active ingredient. Formula (I) below

(wherein any one of R ¹ to R ⁴ represents COR ⁸ , and the other three of R ¹ to R ⁴ are the same or different, hydrogen atom, halogen atom, OH, NO ₂ , CN, alkyl, cycloalkyl, alkoxy, alkylcarbonylamino, alkylcarbonyloxy, arylcarbonyloxy, carbamoyl, arylcarbonylamino, aryl, aralkyl, alkylcarbonyl, SH or alkylthio R 1 and R ² , ^R 2 ^and R ³ , R ³ and R ⁴ represent methylenedioxy, or together with the carbon atoms to which they are attached, a cyclopentene ring, cyclopentadiene ring, cyclohexene ring, cyclohexadiene ring, benzene ring, or a 5-membered or A 6-membered heterocycle may be formed.
R5 represents a hydrogen atom, a halogen atom, alkyl, ^alkoxy , aryl, aralkyl, hydroxyalkyl or cycloalkyl. R ⁴ and R ⁵ together with the carbon atom to which they are bonded may form an optionally substituted benzene ring or an optionally substituted pyridine ring.
R ⁶ and R ⁷ are the same or different and represent a hydrogen atom, alkyl, alkoxy, aryl, aralkyl, hydroxyalkyl or cycloalkyl. However, R6 ^and R7 ^are not hydrogen atoms at the same time.
R ⁸ represents OH, alkoxy, hydroxyalkyloxy, cycloalkyloxy, aryloxy or aralkyloxy.
Z represents N or ^CR9 . R9 represents a hydrogen atom, alkyl, aryl or aralkyl ^.
n represents an integer of 0-5. )
An antidepressant comprising a compound represented by or a pharmaceutically acceptable salt or solvate thereof as an active ingredient.

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