JP2023117263A - glutaminase 1 inhibitor - Google Patents

Abstract

To provide: a glutaminase 1 inhibitor; a pharmaceutical composition, a food and beverage composition or a cosmetic composition containing the inhibitor as an active ingredient; and a new compound useful as an active ingredient of a glutaminase 1 inhibitor.SOLUTION: The glutaminase 1 inhibitor contains as an active ingredient a compound represented by the general formula (I) in the figure, a pharmaceutically acceptable salt thereof, or a solvate of them.SELECTED DRAWING: None

Description

TECHNICAL FIELD The present invention relates to a glutaminase 1 inhibitor, and a pharmaceutical composition, food and drink composition, or cosmetic composition containing the same as an active ingredient. The present invention also relates to novel compounds useful as active ingredients of glutaminase-1 inhibitors.

Glutaminase is an enzyme that produces glutamic acid from glutamine. Glutaminases include kidney-type glutaminase expressed in the kidney (referred to as "glutaminase 1") and liver-type glutaminase expressed in the liver (referred to as "glutaminase 2"). Hereinafter, glutaminase 1 may be abbreviated as "GLS1", glutaminase 2 as "GLS2", and both may be collectively referred to as "GLS".

GLS1 is widely present in the kidney, small intestine, brain, etc., and GLS2 is present only in the liver. Glutamine is the most abundant free amino acid in the body and is known to play a role in regulating many processes such as metabolism, protein synthesis and degradation, and insulin resistance (Non-Patent Document 1). In addition, it has been found that GLS is overexpressed in various cancer cells, and GLS inhibitors such as CB-839 targeting this have been developed as pharmaceuticals (Non-Patent Document 2). In addition, GLS inhibitors such as CB-839 have anti-inflammatory effects and are known to be useful as anti-inflammatory agents and for the prevention, amelioration, or treatment of diseases caused by inflammation (Patent Document 1). . Furthermore, the GLS inhibitor has an anti-obesity effect and is known to be useful as an anti-obesity agent for preventing, improving or treating metabolic syndrome (Patent Document 2). Furthermore, recent research suggests that inhibition of GLS1 has an anti-aging effect (Non-Patent Document 3).

Thus, the importance of GLS as a drug target is increasing today, and new GLS inhibitors are required.

JP 2020-29412 A Japanese Patent Application Laid-Open No. 2020-28239

Molecular mechanisms of glutamine action, J. Cell. Physiol., 2005, 204(2): 392-401. Antitumor activity of the Glutaminase inhibitor CB-839 in triple-negative breast cancer. Mol Cancer Ther. 2014 Apr; 13 (4):890-901. Senolysis by Glutaminolysis inhibition ameliorates various age-associated disorders. Science. 2021 Jan 15; 371 (6526):265-270.

An object of the present invention is to provide a GLS1 inhibitor.
Another object of the present invention is to provide pharmaceutical compositions, food and drink compositions, or cosmetic compositions that are useful for preventing or improving diseases and pathological conditions by inhibiting GLS1.
A further object of the present invention is to provide a novel compound useful as a GLS1 inhibitor or an active ingredient of the pharmaceutical compositions, food and drink compositions, or cosmetic compositions described above.

In order to solve the above problems, the present inventors conducted multistep in silico screening (primary screening using a three-dimensional pharmacophore obtained from the complex crystal structure of glutaminase and a known GLS1 inhibitor, Lipinski's "Rule of Five filtering, secondary screening using molecular docking calculations, filtering by binding free energy calculations, filtering based on docking poses, and molecular similarity analysis based on chemical structures. As a result of searching for inhibitor candidate compounds, it was found that the compound represented by the following formula has GLS1 inhibitory action.

Based on the above findings, the present invention has been completed as a result of repeated studies to discover compounds having GLS1 inhibitory activity similar to the above compounds, and has the following embodiments.

〔式（Ｉ）中、
Ｘは、ＣＨ_２またはＣ＝Ｏ；
ｎは０～２の整数を意味する。但し、ＸがＣ＝Ｏであるとき、ｎは１である。
Ｙは、少なくとも２つの窒素原子を有する二価の５員複素環基；
Ｒ^１は、水素原子、ハロゲン原子、ハロアルキル基、アルコキシ基、アルキル基、または水酸基；
Ｒ^２は、下式（ａ）～（ｃ）のいずれかで示される基：

（式（ａ）中、
Ａは、水酸基または酸素原子を意味し、
点線と実線からなる二重線は、Ａが水酸基である場合は単結合を、酸素原子である場合は二重結合を意味し、
アスタリスクは、式（Ｉ）中のカルボニル基の炭素原子との結合部を意味する。）；

（式（ｂ）中、
Ｂ及びＤは、同一又は異なって、水素原子、アルキル基、又はアルコキシ基を意味し、
アスタリスクは、式（Ｉ）中のカルボニル基の炭素原子との結合部を意味する。）；

（式（ｃ）中、
Ｅは、酸素原子、又は水素原子の一つが水酸基で置換されていてもよいアルキレン基を意味し、
アスタリスクは、式（Ｉ）中のカルボニル基の炭素原子との結合部を意味する。）；
ｎが０である場合、Ｒ_１はアルコキシ基ではなく、また
Ｒ_１が水酸基である場合、Ｒ^２は式（ｂ）または（ｃ）で示される基である）。 (I) GLS1 inhibitor (I-1) A GLS1 inhibitor comprising a compound represented by general formula (I), a pharmaceutically acceptable salt thereof, or a solvate thereof as an active ingredient:

[In formula (I),
X is _CH2 or C=O;
n means an integer of 0-2. However, n is 1 when X is C=O.
Y is a divalent 5-membered heterocyclic group having at least two nitrogen atoms;
R ¹ is a hydrogen atom, a halogen atom, a haloalkyl group, an alkoxy group, an alkyl group, or a hydroxyl group;
R ² is a group represented by any one of the following formulas (a) to (c):

(In formula (a),
A means a hydroxyl group or an oxygen atom,
A double line consisting of a dotted line and a solid line means a single bond when A is a hydroxyl group and a double bond when it is an oxygen atom,
The asterisk denotes the point of attachment to the carbon atom of the carbonyl group in formula (I). );

(In formula (b),
B and D are the same or different and represent a hydrogen atom, an alkyl group, or an alkoxy group;
The asterisk denotes the point of attachment to the carbon atom of the carbonyl group in formula (I). );

(In formula (c),
E is an oxygen atom or an alkylene group in which one of the hydrogen atoms may be substituted with a hydroxyl group,
The asterisk denotes the point of attachment to the carbon atom of the carbonyl group in formula (I). );
When n is 0, R ₁ is not an alkoxy group, and when R ₁ is a hydroxyl group, R ² is a group represented by formula (b) or (c)).

(I-2) The GLS1 inhibitor according to (I-1), wherein in the compound represented by formula (I) above, Y is a pyrazole group or an imidazole group, preferably a pyrazole group.

(I-3) the compound represented by the formula (I), wherein R ² is a group represented by the formula (a),
In formula (a), A is a hydroxyl group, a double line consisting of a dotted line and a solid line is a single bond,
The GLS1 inhibitor according to (I-1) or (I-2), wherein X is CH ₂ and n is an integer of 0-2.
(I-4) the compound represented by the formula (I), wherein R ² is a group represented by the formula (b),
The GLS1 inhibitor according to (I-1) or (I-2), wherein X is CH ₂ and n is an integer of 0-2.
(I-5) the compound represented by the formula (I), wherein R ² is a group represented by the formula (c),
The GLS1 inhibitor according to (I-1) or (I-2), wherein X is CH ₂ and n is 0-2.

(II) Pharmaceutical composition, food and drink composition, or cosmetic composition (II-1) A pharmaceutical composition containing the GLS1 inhibitor according to any one of (I-1) to (I-5) as an active ingredient , food and beverage compositions, or cosmetic compositions.
(II-2) The pharmaceutical composition, food and drink composition according to (II-1), which is used for the prevention, improvement or treatment of inflammation, diseases derived from inflammation, obesity, metabolic syndrome, aging, or cancer. product or cosmetic composition.

〔式（Ｉ）中、
Ｘは、ＣＨ_２またはＣ＝Ｏ；
ｎは０～２の整数を意味する。但し、ＸがＣ＝Ｏであるとき、ｎは１である。
Ｙは、少なくとも２つの窒素原子を有する二価の５員複素環基；
Ｒ^１は、水素原子、ハロゲン原子、ハロアルキル基、アルコキシ基、アルキル基、または水酸基；
Ｒ^２は、下式（ａ）～（ｃ）のいずれかで示される基：

（式（ａ）中、
Ａは、水酸基または酸素原子を意味し、
点線と実線からなる二重線は、Ａが水酸基である場合は単結合を、酸素原子である場合は二重結合を意味し、
アスタリスクは、式（Ｉ）中のカルボニル基の炭素原子との結合部を意味する。）；

（式（ｂ）中、
Ｂ及びＤは、同一又は異なって、水素原子、アルキル基、又はアルコキシ基を意味し、
アスタリスクは、式（Ｉ）中のカルボニル基の炭素原子との結合部を意味する。）；

（式（ｃ）中、
Ｅは、酸素原子、又は水素原子の一つが水酸基で置換されていてもよいアルキレン基を意味し、
アスタリスクは、式（Ｉ）中のカルボニル基の炭素原子との結合部を意味する。）；
ｎが０である場合、Ｒ_１はアルコキシ基ではなく、また
Ｒ_１が水酸基である場合、Ｒ^２は式（ｂ）または（ｃ）で示される基である。
但し、Ｎ－（４－ヒドロキシシクロヘキシル）－３－フェニル－１Ｈ－ピラゾール－５－カルボキサミドを除く。〕。 (III) Compounds having GLS1 inhibitory activity (III-1) Compounds having GLS1 inhibitory activity represented by general formula (I), pharmaceutically acceptable salts thereof, or solvates thereof:

[In formula (I),
X is _CH2 or C=O;
n means an integer of 0-2. However, n is 1 when X is C=O.
Y is a divalent 5-membered heterocyclic group having at least two nitrogen atoms;
R ¹ is a hydrogen atom, a halogen atom, a haloalkyl group, an alkoxy group, an alkyl group, or a hydroxyl group;
R ² is a group represented by any one of the following formulas (a) to (c):

(In formula (a),
A means a hydroxyl group or an oxygen atom,
A double line consisting of a dotted line and a solid line means a single bond when A is a hydroxyl group and a double bond when it is an oxygen atom,
The asterisk denotes the point of attachment to the carbon atom of the carbonyl group in formula (I). );

(In formula (b),
B and D are the same or different and represent a hydrogen atom, an alkyl group, or an alkoxy group;
The asterisk denotes the point of attachment to the carbon atom of the carbonyl group in formula (I). );

(In formula (c),
E is an oxygen atom or an alkylene group in which one of the hydrogen atoms may be substituted with a hydroxyl group,
The asterisk denotes the point of attachment to the carbon atom of the carbonyl group in formula (I). );
When n is 0, R ₁ is not an alkoxy group, and when R ₁ is a hydroxyl group, R ² is a group represented by formula (b) or (c).
However, N-(4-hydroxycyclohexyl)-3-phenyl-1H-pyrazole-5-carboxamide is excluded. ].

(III-2) The compound according to (III-1), wherein Y is a pyrazole group or an imidazole group, preferably a pyrazole group.

(III-3) R ² is a group represented by formula (a),
In formula (a), A is a hydroxyl group, a double line consisting of a dotted line and a solid line is a single bond,
The compound according to (III-1) or (III-2), wherein X is CH ₂ and n is an integer of 0-2.
(III-4) the compound represented by the formula (I), wherein R ² is a group represented by the formula (b),
The compound according to (III-1) or (III-2), wherein X is CH ₂ and n is an integer of 0-2.
(III-5) The compound represented by the formula (I), wherein R ² is a group represented by the formula (c),
The compound according to (III-1) or (III-2), wherein X is CH ₂ and n is 0-2.

According to the present invention, a novel GLS1 inhibitor can be provided. The GLS1 inhibitor of the present invention is a drug for preventing or improving diseases and pathological conditions (e.g., inflammation, diseases and pathological conditions derived from inflammation, obesity, metabolic syndrome, aging, cancer, etc.) that are successful by inhibiting GLS1. It is useful as an active ingredient of compositions, food and drink compositions, or cosmetic compositions.
Therefore, the pharmaceutical composition, food and drink composition, or cosmetic composition of the present invention, which contains a GLS1 inhibitor as an active ingredient, inhibits GLS1, thereby inhibiting, for example, inflammation, inflammation-derived diseases and conditions, obesity, and metabolic syndrome. It is useful as having an effect of preventing or improving syndrome, aging, cancer, or the like.
Furthermore, by inhibiting GLS1, the compounds provided by the present invention can exert an effect of preventing or improving inflammation, diseases or conditions derived from inflammation, obesity, metabolic syndrome, aging, cancer, or the like. It is expected and useful as the GLS1 inhibitor and an active ingredient of pharmaceutical compositions, food and drink compositions, or cosmetic compositions.

The structural formulas of GLS1 inhibitory candidate compounds evaluated for GLS1 inhibitory activity in Experimental Examples are shown. The structural formulas of GLS1 inhibitory candidate compounds evaluated for GLS1 inhibitory activity in Experimental Examples are shown. The structural formulas of GLS1 inhibitory candidate compounds evaluated for GLS1 inhibitory activity in Experimental Examples are shown. (A) shows the structural formulas of GLS1 inhibitory candidate compounds 1 to 8 evaluated for GLS1 inhibitory activity in Experimental Example 1; (B) For GLS1 inhibitor candidate compounds 1 to 8, known GLS1 inhibitors (DON, CB-839), and control (DMSO), glutamic acid (Glu) production rate (%) was measured using mouse kidney extract. Show the results. 2 shows the results of measurement of the Glu production rate (%) using mouse kidney extract for GLS1 inhibitory candidate compounds, known GLS1 inhibitors (DON, CB-839), and control (DMSO) in Experimental Example 2. FIG. In Experimental Example 3, the GLS1 inhibitory candidate compounds (pyrazole derivatives 5i and 8a), the known GLS1 inhibitors (DON, CB-839), and the control (DMSO) were evaluated for the glutamic acid production rate (%) using mouse recombinant GLS1. Measured results are shown. P value was calculated by Tukey-Kramer HSD test. n=3 *P<0.05 (vs Ct). In Experimental Example 3, the GLS1 inhibitory candidate compounds (pyrazole derivatives 5i and 8a), known GLS1 inhibitors (DON, CB-839), and control (DMSO) were evaluated for the glutamic acid production rate (%) using human recombinant GLS1. Measured results are shown. P value was calculated by Tukey-Kramer HSD test. n=3 *P<0.05 (vs Ct). 4 shows the results of evaluating the growth inhibitory activity of pyrazole derivatives 5i and 8a on human mammary adenocarcinoma-derived cell lines (MCF7 cells) in Experimental Example 4. FIG. n=3 4 shows the results of evaluating the antiproliferative effects of pyrazole derivatives 5i and 8a on acute myelogenous leukemia-derived cell lines (MOLM13 cells) in Experimental Example 4. FIG. n=3

(I) GLS1 inhibitor The GLS1 inhibitor of the present invention is a compound represented by the following formula (I) (hereinafter also referred to as "compound (I)"), a pharmaceutically acceptable salt thereof, or Characterized by having a solvate as an active ingredient:

In formula (I), X represents CH ₂ or C=O. _CH2 is preferred.
When X is CH ₂ , n is an integer from 0-2. Preferably 0 or 1. n is 1 when X is C=O.

In formula (I), Y is a divalent 5-membered heterocyclic group having at least two nitrogen atoms (hereinafter also simply referred to as "N-containing 5-membered heterocyclic group"). The N-containing 5-membered heterocyclic group may be a divalent 5-membered heterocyclic group having 2 or 3 nitrogen atoms in the ring and an atom bonding to the adjacent group is a carbon atom, but preferably It is an N-containing 5-membered unsaturated heterocyclic group. The N-containing 5-membered heterocyclic group may have a carbon atom, or a heteroatom such as an oxygen atom or a sulfur atom, in addition to the nitrogen atom, as a group other than the carbon atom serving as a bond. For example, N-containing 5-membered unsaturated heterocyclic groups in which the atoms other than the two nitrogen atoms are carbon atoms include a pyrazole group and an imidazole group; Examples of saturated heterocyclic groups include furazane groups; and examples of N-containing 5-membered unsaturated heterocyclic groups having 3 nitrogen atoms include triazole groups. The N-containing 5-membered heterocyclic group is preferably a pyrazole group in which atoms other than two nitrogen atoms are carbon atoms or an imidazole group, more preferably a pyrazole group.

It is preferable that the two bonds of the N-containing 5-membered heterocyclic group are positioned as far apart as possible. For example, when the position of the bond of the N-containing 5-membered heterocyclic group bonded to the X group that is part of the main skeleton of the compound represented by formula (I) is the 1-position, the other bond is It is preferably located at the 3rd or 4th position.

In formula (I), R ¹ is a hydrogen atom, a halogen atom, a haloalkyl group, an alkoxy group, an alkyl group, or a hydroxyl group. Although not limited, hydrogen atoms, halogen atoms, haloalkyl groups and alkoxy groups are preferred, and hydrogen atoms and halogen atoms are more preferred.
In formula (I), when n is 0, R ₁ is preferably a group other than an alkoxy group. More preferred are a hydrogen atom, a halogen atom and a hydroxyl group.
Moreover, when R ₁ is a hydroxyl group, R ² is preferably a group represented by formula (b) or (c) described later. Groups represented by formula (b) are more preferred.

Halogen atoms represented by R ¹ include, for example, fluorine, chlorine, bromine and iodine atoms. A fluorine atom and a chlorine atom are preferred.

The haloalkyl group represented by R ¹ includes a linear or branched alkyl group having 1 to 6 carbon atoms (C1 to 6) and having 1 to 3 halogen atoms. A trihalo C1-6 alkyl group having 3 halogen atoms is preferred, and a trihalo C1-3 alkyl group is more preferred. Trihalo C1-6 alkyl groups include, for example, trifluoromethyl group, trichloromethyl group, 2,2,2-trifluoroethyl group, 2,2,2-trichloroethyl group and 3,3,3-trifluoropropyl group. And so on. A trifluoromethyl group is preferred.

The alkyl group represented by R ¹ is not limited, but preferably includes a linear or branched alkyl group having 1 to 6 carbon atoms (C1 to 6). More preferably, it is a C1-3 alkyl group. These alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, and n-hexyl groups. Although not limited, methyl group, ethyl group and tert-butyl group are preferred, and methyl group is more preferred.

The alkoxy group represented by R ¹ is a group having a structure in which an alkyl group is bonded to an oxygen atom. The alkyl group contained in this structure is also as described above, preferably a C1-3 alkyl group. Alkoxy groups specifically include methoxy and ethoxy groups. A methoxy group is preferred.

In formula (I), the position of R ¹ on the phenyl group may be ortho, meta or para to the X group attached to the benzene ring. Without limitation, when n is 0, it is preferably in the para position. Also, when n is 1, the para-position, the meta-position, and the ortho-position are preferred in that order, and the para-position is more preferred.

式（ａ）中、Ａは、水酸基または酸素原子を意味する。
Ａが水酸基である場合、式（ａ）で示す点線と実線からなる二重線は、単結合を意味する。
Ａが酸素原子である場合、式（ａ）で示す点線と実線からなる二重線は、二重結合を意味する。
式（ａ）中、アスタリスクは、前記式（Ｉ）中のカルボニル基の炭素原子との結合部を意味する。 In formula (I), R ² is a group represented by any one of formulas (a) to (c) shown below.
A group represented by formula (a) :

In formula (a), A means a hydroxyl group or an oxygen atom.
When A is a hydroxyl group, the double line consisting of a dotted line and a solid line in formula (a) means a single bond.
When A is an oxygen atom, the double line consisting of a dotted line and a solid line in formula (a) means a double bond.
In formula (a), an asterisk means a bond to the carbon atom of the carbonyl group in formula (I).

式（ｂ）中、Ｂ及びＤは、同一又は異なって、水素原子、アルキル基、又はアルコキシ基を意味する。
ここでアルキル基としては、前述するＲ^１で示されるアルキル基と同じく、好ましくはＣ１～６の直鎖状または分岐状のアルキル基を挙げることができる。より好ましくはＣ１～３の直鎖状アルキル基であり、具体的にはメチル基、及びエチル基を挙げることができる。
アルコキシ基としても、前述するＲ^１で示されるアルコキシ基と同じく、Ｃ１～３のアルキル基を有するアルコキシ基を好適に挙げることができる、好ましくはメトキシ基及びエトキシ基であり、より好ましくはメトキシ基である。
ＢとＤの組み合わせには、制限されないものの、水素原子とアルキル基の組み合わせ、アルキル基同士の組み合わせ、アルキル基とアルコキシ基との組み合わせ、水素原子とアルコキシ基の組み合わせが含まれる。
式（ｂ）中、アスタリスクは、式（Ｉ）中のカルボニル基の炭素原子との結合部を意味する。 Group represented by formula (b) :

In formula (b), B and D are the same or different and mean a hydrogen atom, an alkyl group, or an alkoxy group.
Here, the alkyl group is preferably a C1-6 linear or branched alkyl group, like the alkyl group represented by R ¹ described above. More preferably, it is a C1-3 linear alkyl group, and specific examples include a methyl group and an ethyl group.
As the alkoxy group, similarly to the alkoxy group represented by R ¹ described above, an alkoxy group having a C1-3 alkyl group can be preferably mentioned, preferably a methoxy group and an ethoxy group, more preferably a methoxy group. is.
Combinations of B and D include, but are not limited to, combinations of hydrogen atoms and alkyl groups, combinations of alkyl groups, combinations of alkyl groups and alkoxy groups, and combinations of hydrogen atoms and alkoxy groups.
In formula (b), the asterisk means the bond to the carbon atom of the carbonyl group in formula (I).

式（ｃ）中、Ｅは、酸素原子、又はアルキレン基を意味する。アルキレン基には、制限されないものの、炭素数１～３、好ましくは炭素数１～２のアルキレン基が含まれる。当該アルキレン基はその水素原子の一つが水酸基で置換されていてもよい。
式（ｃ）中、アスタリスクは、前記式（Ｉ）中のカルボニル基の炭素原子との結合部を意味する。 Group represented by formula (c) :

In formula (c), E means an oxygen atom or an alkylene group. Alkylene groups include, but are not limited to, those having 1 to 3 carbon atoms, preferably 1 to 2 carbon atoms. One of the hydrogen atoms in the alkylene group may be substituted with a hydroxyl group.
In formula (c), an asterisk means a bond to the carbon atom of the carbonyl group in formula (I).

Compound (I) includes the following compounds as preferred compounds:
(A) Compounds in which R2 ^is a group represented by formula (a) (collectively referred to as "compound a"):
Compound a includes the following compounds.
(a1) In formula (a), A is a hydroxyl group; a double line consisting of a dotted line and a solid line is a single bond; X is CH ₂ ; n is an integer of 0 to 2; a pyrazole group; a compound in which ^R1 is a hydrogen atom, a halogen atom, a haloalkyl group, an alkoxy group, or an alkyl group;
Among them, preferably, in formula (a), A is a hydroxyl group; a double line consisting of a dotted line and a solid line is a single bond; X is CH ₂ ; n is 0 or 1; Pyrazole group; a compound in which R ¹ is a hydrogen atom or a halogen atom.

These compounds specifically include the following compounds. All such compounds also include isomers, regardless of whether they are stereoisomers.
Compound 1: N-(4-hydroxycyclohexyl)-3-phenyl-1H-pyrazole-5-carboxamide (N-(4-hydroxycyclohexyl)-3-phenyl-1H-pyrazole-5-carboxamide)
Compound 2: 3-(4-fluorophenyl)-N-(4-hydroxycyclohexyl)-1H-pyrazole-5-carboxamide (3-(4-fluorophenyl)-N-(4-hydroxycyclohexyl)-1H-pyrazole-5 -carboxamide)
Compound 3: 3-benzyl-N-(4-hydroxycyclohexyl)-1H-pyrazole-5-carboxamide
Compound 4: 3-(4-fluorobenzyl)-N-(4-hydroxycyclohexyl)-1H-pyrazole-5-carboxamide (3-(4-fluorobenzyl)-N-(4-hydroxycyclohexyl)-1H-pyrazole-5 -carboxamide)
Compound 5: 3-(4-chlorobenzyl)-N-(4-hydroxycyclohexyl)-1H-pyrazole-5-carboxamide (3-(4-chlorobenzyl)-N-(4-hydroxycyclohexyl)-1H-pyrazole-5 -carboxamide)
Compound 6: 3-(2-chlorobenzyl)-N-(4-hydroxycyclohexyl)-1H-pyrazole-5-carboxamide (3-(2-chlorobenzyl)-N-(4-hydroxycyclohexyl)-1H-pyrazole-5 -carboxamide)
Compound 7: 3-(3-chlorobenzyl)-N-(4-hydroxycyclohexyl)-1H-pyrazole-5-carboxamide (3-(3-chlorobenzyl)-N-(4-hydroxycyclohexyl)-1H-pyrazole-5 -carboxamide)
Compound 8: N-(4-hydroxycyclohexyl)-3-(4-(trifluoromethyl)benzyl)-1H-pyrazole-5-carboxamide (N-(4-hydroxycyclohexyl)-3-(4-(trifluoromethyl)benzyl )-1H-pyrazole-5-carboxamide)
Compound 9: N-(4-hydroxycyclohexyl)-3-(3-methoxybenzyl)-1H-pyrazole-5-carboxamide (N-(4-hydroxycyclohexyl)-3-(3-methoxybenzyl)-1H-pyrazole-5 -carboxamide)
Compound 10: N-(4-hydroxycyclohexyl)-3-(4-methoxybenzyl)-1H-pyrazole-5-carboxamide (N-(4-hydroxycyclohexyl)-3-(4-methoxybenzyl)-1H-pyrazole-5 -carboxamide)
Compound 11: N-(4-hydroxycyclohexyl)-3-(2-methoxybenzyl)-1H-pyrazole-5-carboxamide (N-(4-hydroxycyclohexyl)-3-(2-methoxybenzyl)-1H-pyrazole-5 -carboxamide)
Compound 12: N-(4-hydroxycyclohexyl)-3-(4-methylbenzyl)-1H-pyrazole-5-carboxamide (N-(4-hydroxycyclohexyl)-3-(4-methylbenzyl)-1H-pyrazole-5 -carboxamide)
Compound 13: N-(4-hydroxycyclohexyl)-3-phenethyl-1H-pyrazole-5-carboxamide

(a2) In formula (a), A is an oxygen atom; a double line consisting of a dotted line and a solid line is a double bond; X is CH ₂ ; n is 0 to 2, preferably 1; a pyrazole group having a bond; ^R1 is a hydrogen atom;
The compound specifically includes the following compounds. All such compounds also include isomers, regardless of whether they are stereoisomers.
Compound 14: 3-benzyl-N-(4-oxocyclohexyl)-1H-pyrazole-5-carboxamide

(a3) In formula (a), A is a hydroxyl group; the double line consisting of a dotted line and a solid line is a single bond; X is C=O; n is 1; A compound wherein R ¹ is a hydrogen atom.
The compound specifically includes the following compounds. All such compounds also include isomers, regardless of whether they are stereoisomers.
Compound 15: 3-benzoyl-N-(4-hydroxycyclohexyl)-1H-pyrazole-5-carboxamide

(a4) In formula (a), A is a hydroxyl group; a double line consisting of a dotted line and a solid line is a single bond; X is CH ₂ ; n is 0 to 2, preferably 1; an imidazole group having a; R ¹ is a hydrogen atom.
The compound specifically includes the following compounds. All such compounds also include isomers, regardless of whether they are stereoisomers.
Compound 16: N-(4-hydroxycyclohexyl)-5-phenyl-1H-imidazole-2-carboxamide

(B) Compounds in which R2 ^is a group represented by formula (b) (collectively referred to as "compound b"):
Compound b includes the following compounds.
(b) in formula (b), B and D are the same or different and are a hydrogen atom, an alkyl group, or an alkoxy group; X is CH ₂ ; n is an integer of 0 to 2; a pyrazole group having a chain; ^R1 is a hydrogen atom or a hydroxyl group.
The compound specifically includes the following compounds. All such compounds also include isomers, regardless of whether they are stereoisomers.
Compound 17: 3-(4-hydroxyphenyl)-N-methyl-1H-pyrazole-5-carboxamide
Compound 18: 3-benzyl-N-methyl-1H-pyrazole-5-carboxamide
Compound 19: 3-benzyl-N,N-dimethyl-1H-pyrazole-5-carboxamide
Compound 20: 3-benzyl-N-methoxy-N-methyl-1H-pyrazole-5-carboxamide

(C) Compounds in which R2 ^is a group represented by formula (c) (collectively referred to as "compound c"):
Compound c includes the following compounds.
(c) In formula (c), E is an oxygen atom or an alkylene group in which one hydrogen atom may be substituted with a hydroxyl group; X is CH ₂ ; n is 0 to 2; Y is 3- and 5-positions A compound wherein a pyrazole group having a bond to ^R1 is a hydrogen atom.
The compound specifically includes the following compounds. All such compounds also include isomers, regardless of whether they are stereoisomers.
Compound 21: (3-benzyl-1H-pyrazol-5-yl)(piperidin-1-yl)methanone
Compound 22: Azepan-1-yl(3-benzyl-1H-pyrazol-5-yl)methanone
Compound 23: (3-benzyl-1H-pyrazol-5-yl)(morpholino)methanone
Compound 24: (3-benzyl-1H-pyrazol-5-yl)(4-hydroxypiperidin-1-yl)methanone )

Compound (I) described above can take the form of a pharmaceutically acceptable salt, and the salt can be used in the same manner as compound (I) in free form. Pharmaceutically acceptable salts include salts with inorganic acids such as hydrochloric, phosphoric, sulfuric, nitric, or boric acids (mineral acid salts); formic, acetic, lactic, fumaric, maleic, tartaric, citric, Acids, salts with organic acids such as succinic acid, malonic acid, or tosylic acid (organic acid salts) are included. However, it is not limited to these. Preparation of these salts can be carried out by conventional means. In addition, the aforementioned compound (I) and its pharmaceutically acceptable salts may be solvates. Solvates also include hydrates.

Compound (I), a pharmaceutically acceptable salt thereof, or a solvate thereof (hereinafter collectively referred to as "compound (I), etc.") is produced based on the description of Production Examples described later. can do.

Compound (I) and the like obtained by these production methods are isolated and purified from the reaction mixture by applying known isolation and/or purification means. Examples of such isolation and purification means include distillation, recrystallization, solvent extraction, column chromatography, ion exchange chromatography, gel chromatography, affinity chromatography, preparative thin layer chromatography, and the like. can be done.

Compound (I) and the like have GLS1 inhibitory activity as shown in Experimental Examples 2 and 3 below.

The GLS1 inhibitor according to the present invention may consist of only the above-described compound (I), etc., but is commonly used in the relevant fields (biochemical field, pharmaceutical field, cosmetic field, food field). Other ingredients such as carriers and additives may be included. The specific types of carriers and additives and their mixing ratios are not particularly limited as long as the GLS1 inhibitor exhibits GLS1 inhibitory activity based on compound (I) and the like. For example, it can be appropriately selected with reference to the types and blending ratios of carriers and additives described in the pharmaceutical composition described later.

(II) Pharmaceutical composition The pharmaceutical composition according to the present invention (hereinafter referred to as "the pharmaceutical composition") comprises the GLS1 inhibitor described above, specifically compound (I), a pharmaceutically acceptable or a solvate thereof as an active ingredient.

Based on the GLS1 inhibitory activity of compound (I) and the like described above, the pharmaceutical composition can be applied to diseases or symptoms (including conditions) that are effective by inhibiting GLS1. Diseases or conditions to be treated with the present pharmaceutical composition include inflammation, diseases derived from inflammation, obesity, metabolic syndrome, aging, and cancer.

The pharmaceutical composition can be suitably used for prevention or treatment of such diseases or symptoms. In the present invention, treatment includes not only the meaning of curing the above diseases and symptoms, but also alleviation and partial amelioration. In the present invention, prevention also includes suppression of disease or symptom progression and suppression of recurrence. In particular, the prevention or treatment of cancer includes the reduction of cancer tissue, the prevention and suppression of cancer progression, and the suppression of cancer recurrence by suppressing the growth of cancer cells.

In the present invention, the term "cancer" is interpreted broadly and used interchangeably with the term "malignant tumor". In addition, in the stage before the diagnosis is confirmed pathologically, that is, before the tumor is either benign or malignant, it may include benign tumors, benign-malignant borderline lesions, and malignant tumors collectively. could be. In general, cancers are called by the name of the organ from which they originated, or by the name of the originating tissue. cancer, salivary gland cancer, esophageal cancer, stomach cancer, small intestine cancer, colon cancer, rectal cancer, liver cancer, biliary tract cancer, gallbladder cancer, pancreatic cancer, lung cancer, breast cancer, thyroid cancer cancer, adrenal cancer, pituitary tumor, pineal tumor, uterine cancer, ovarian cancer, vaginal cancer, bladder cancer, kidney cancer, prostate cancer, urethral cancer, retinoblastoma, conjunctiva Cancer, neuroblastoma, glioma, glioblastoma, skin cancer, medulloblastoma, leukemia, malignant lymphoma, testicular tumor, osteosarcoma, rhabdomyosarcoma, leiomyosarcoma, angiosarcoma, liposarcoma , chondrosarcoma, Ewing's sarcoma, multiple myeloma, acute myelogenous leukemia, and the like. Furthermore, depending on the characteristics of the site of the organ in which it develops, it can be classified as upper/middle/hypopharynx cancer, upper/middle/lower esophageal cancer, gastric cardia cancer, gastric pyloric cancer, cervical cancer, endometrial cancer, etc. Subclassifications include, but are not limited to, the description of "cancer" in the present invention. The pharmaceutical composition of the present invention used for anticancer therapy (also referred to as "anticancer pharmaceutical composition") is effective against "cancer" in general, but is particularly effective against breast cancer and acute cancer. It can be preferably used for myeloid leukemia. In particular, among compound (I) and the like, compounds 1 and 4, and pharmaceutically acceptable salts and solvates thereof, have anticancer effects, particularly in breast cancer and/or acute myeloid leukemia, cancer It has an excellent cell growth inhibitory effect.

The term "anti-cancer pharmaceutical composition" refers to a pharmaceutical composition that exhibits a therapeutic or preventive effect on cancer, which is the target disease or condition, but preferably a pharmaceutical composition that exhibits a therapeutic effect. refers to things. Therapeutic effects include alleviation of symptoms characteristic of cancer or accompanying symptoms (mitigation), prevention or delay of exacerbation of symptoms, and the like. The latter can be regarded as one of preventive effects in terms of preventing aggravation. Thus, the therapeutic effect and the prophylactic effect are partially overlapping concepts, and it is difficult to clearly distinguish between them, and there is little practical benefit from doing so. A typical preventive effect is to prevent or delay the recurrence (development) of symptoms characteristic of cancer. In addition, as long as it shows some therapeutic effect or preventive effect, or both, against cancer, it corresponds to the anticancer pharmaceutical composition. In addition, the therapeutic or preventive effect on cancer brought about by compound (I) and the like may include improvement of cancer complications (eg, cachexia). That is, the terms "anticancer" and "cancer treatment" as used herein refer to effects such as growth suppression and shrinkage on cancer tissue itself, as well as complications (preferably, cachexia). ).
The "anticancer pharmaceutical composition" of the present invention can be used in combination with known anticancer agents or anticancer agents that will be developed in the future.

The present pharmaceutical composition may consist solely of compound (I) and the like described above, or may be combined with other pharmaceutically acceptable ingredients and administered according to a desired administration route, administration method, etc., by a known method. It may be prepared in a form suitable for the application. Formulation of the present pharmaceutical composition can be carried out according to a conventional method, except that compound (I) and the like, which are active ingredients, are blended.
When formulating, other pharmaceutically acceptable components (e.g., carriers, excipients, disintegrants, buffers, emulsifiers, suspending agents, soothing agents, stabilizers, preservatives, preservatives, interfaces Active agents, lubricants, diluents, coating agents, sugar coating agents, flavoring agents, emulsifying/solubilizing/dispersing agents, pH adjusters, isotonic agents, solubilizing agents, fragrances, coloring agents, solubilizing agents, physiology saline solution, etc.).
The dosage form for formulation is also not particularly limited. Dosage forms include tablets, powders, fine granules, granules, capsules, syrups, solutions, suspensions, emulsions, jellies, injections, external preparations, inhalants, nasal drops, eye drops, and suppositories. drug is included.

The pharmaceutical composition contains, as an active ingredient, compound (I) and the like in an amount necessary to obtain the expected prophylactic or therapeutic effect (that is, a therapeutically effective amount). The amount of compound (I) and the like in the present pharmaceutical composition varies depending on the dosage form, but is, for example, within the range of about 0.01% by mass to about 99.9% by mass so that the desired dosage can be achieved. can be set.

Depending on the dosage form, the pharmaceutical composition can be administered orally or parenterally (intravenous, intraarterial, subcutaneous, intradermal, intramuscular, or intraperitoneal injection, transdermal, nasal, transmucosal, etc.) Applied to the subject. These routes of administration are not mutually exclusive, and two or more arbitrarily selected routes can be used in combination (for example, intravenous injection or the like is performed at the same time as oral administration or after a predetermined period of time has elapsed). Local administration may be used instead of systemic administration. A drug delivery system (DDS) may be used to deliver the active ingredient in a target tissue-specific manner. The "subject" here is not particularly limited, and includes humans and non-human mammals (including pet animals, domestic animals, and experimental animals) that require treatment or prevention. Specifically, for example, mice, rats, guinea pigs, hamsters, monkeys , cows, pigs, goats, sheep, dogs, cats, chickens, quail, etc.). Humans are preferred.

The dose of the present pharmaceutical composition may generally vary depending on the patient's symptoms, age, sex, body weight, etc., but a person skilled in the art can appropriately set the appropriate dose. Although not limited, for oral administration, for example, about 0.01 mg to 1000 mg per day can be administered to an adult once or in several divided doses. In parenteral administration, for example, about 0.01 mg to 1000 mg can be administered by subcutaneous injection, intramuscular injection or intravenous injection. As an administration schedule, for example, once to several times a day, once every two days, or once every three days can be adopted. In setting the administration schedule, the patient's symptoms, duration of effect of the active ingredient, etc. can be taken into consideration.

(III) Food and drink composition The food and drink composition according to the present invention (hereinafter referred to as "the present food and drink composition") comprises the GLS1 inhibitor described above, specifically the compound (I), and its pharmaceutical composition. or a solvate thereof.
The present food and drink composition is not limited, but can be prepared by blending the compound (I) and the like in a conventionally known food and drink manufacturing process. Moreover, it can also be prepared by adding and blending the compound (I) and the like to conventionally known food and drink.
The present food and drink composition can be effectively used for, but not limited to, anti-obesity, prevention or improvement of metabolic syndrome.

(IV) Cosmetic Composition The cosmetic composition according to the present invention (hereinafter referred to as "the present cosmetic composition") comprises the GLS1 inhibitor described above, specifically the compound (I), a pharmaceutically acceptable or a solvate thereof. This cosmetic composition also includes medicated cosmetics that correspond to quasi-drugs.
The present cosmetic composition is not limited, but can be prepared by blending the compound (I) and the like in the manufacturing process of conventionally known cosmetics (including cosmeceuticals). It can also be prepared by adding and blending the compound (I) and the like to conventionally known cosmetic ingredients.
The cosmetic composition is useful for, but not limited to, anti-aging, specifically anti-wrinkle and anti-aging.

As used herein, the terms "include" and "contain" include the meanings of "consisting of" and "consisting essentially of". Moreover, the present invention is not limited to the above-described embodiments, and can be freely modified within the scope of the present invention.

Hereinafter, the present invention will be described using experimental examples in order to facilitate understanding of the configuration and effects of the present invention. However, the present invention is not limited by these experimental examples. Unless otherwise specified, the following experiments were performed at room temperature (25±5° C.) and atmospheric pressure conditions. Unless otherwise specified, "%" described below means "% by mass", and "part" means "parts by mass".

Production Example 1: Synthesis of GLS1 Inhibitor Candidate Compounds GLS1 inhibitor candidate compounds (compounds 1 to 14, 17 to 24, and 3a) were produced according to the procedure shown in the formula below.

(1) Synthesis of compounds 2a-2o
Sodium bis(trimethylsilyl)amide (0.79 mL, 1.9 M in THF, 1.50 mmol) was added to a solution of methyl ketone (1.00 mmol) (compounds 1a to 1o) in tetrahydrofuran (5 mL) at -78°C under Ar atmosphere. After the addition, the mixture was stirred at -78°C for 30 minutes. Furthermore, after adding diethyl oxalate (0.27 mL, 2.00 mmol) at -78°C, the temperature was raised to room temperature, and the mixture was stirred at room temperature for 2 hours. After completion of the reaction, 10% HCl aqueous solution (3 mL) was added. After separating the organic layer, the aqueous layer was extracted with dichloromethane (1.5 mL × 3) and combined with the previous organic layer. After drying the organic layer with anhydrous sodium sulfate, the solvent was distilled off using a rotary evaporator, and the resulting ketoester was used in the next reaction without purification. Under an Ar atmosphere, hydrazine monohydrate (0.07 mL, 1.50 mmol) was added to an ethanol (5 mL) solution of the obtained ketoester at room temperature, and the mixture was stirred under reflux with heating for 4 hours. After completion of the reaction, the reaction solution was diluted with ethyl acetate (5 mL), and water (3 mL) was added. After separating the organic layer, the aqueous layer was extracted with ethyl acetate (1.5 mL × 3) and combined with the previous organic layer. After drying the organic layer with anhydrous sodium sulfate, the solvent was distilled off using a rotary evaporator, and the resulting residue was purified by silica gel column chromatography (n-hexane:ethyl acetate = 7:1 to 1:1). By doing so, the desired pyrazole esters (compounds 2a to 2o) were obtained.

(2) Synthesis of Compounds 1-14 and 18-24 and Compound 3a
Under an Ar atmosphere, 10% NaOH aqueous solution (1.5 mL) was added to a solution of pyrazole ester (compound 2a-2o) (0.50 mmol) in ethanol (3 mL) at room temperature, and the mixture was stirred under reflux for 2 hours. . After completion of the reaction, ethanol was distilled off, 10% HCl aqueous solution (3 mL) was added, and the aqueous layer was extracted with ethyl acetate (1.5 mL x 3). After the organic layer was dried using anhydrous sodium sulfate, the solvent was distilled off using a rotary evaporator, and the resulting carboxylic acid was used in the next reaction without purification. Under Ar atmosphere, the corresponding amine (0.75 mmol), triethylamine (0.11 mL, 0.75 mmol), 1-hydroxybenzotriazole (101 mg, 0.75 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (143 mg, 0.75 mmol) were sequentially added, and the mixture was stirred at room temperature for 20 hours. After completion of the reaction, the solvent was removed using a rotary evaporator, and the resulting residue was purified by silica gel column chromatography (n-hexane:ethyl acetate = 7:1 to 1:5) to give the target compound ( Compounds 1-14 and 18-24 and compound 3a) were obtained.

(3) Synthesis of compound 17
Under an Ar atmosphere, to a methanol (3 mL) solution of amide (compound 3a) (0.50 mmol), 10% HCl aqueous solution (1.5 mL) was added at room temperature, and the mixture was stirred under reflux with heating for 30 minutes. After completion of the reaction, the solvent was removed using a rotary evaporator, and the resulting residue was purified by silica gel column chromatography (n-hexane:ethyl acetate = 1:1 to 1:5) to give the target compound ( Compound 17) was obtained.

Properties of each compound synthesized by the above method are shown below.

Compound 1: N-(trans-4-hydroxycyclohexyl)-3-phenyl-1H- pyrazole-5-carboxamide
¹ H-NMR (400 MHz, CD ₃ OD) δ: 1.37-1.51 (4H, m), 2.00-2.06 (4H, m), 3.58-3.61 (1H, m), 3.83-3.90 (1H, m), 7.02 (1H, s), 7.38-7.47 (3H, m), 7.69-7.79 (2H, m). ^[1]

Compound 2: 3-(4-fluorophenyl)-N-(trans-4-hydroxycyclohexyl)-1H-pyrazole-5-carboxamide
mp: 237-239 °C; ^1H -NMR (400 MHz, _CD3OD ) δ: 1.37-1.51 (4H, m), 1.99-2.02 (4H, m), 3.56-3.58 (1H, m), 3.85- 3.87 (1H, m), 7.00 (1H, s), 7.19 (2H, dr), 7.74 (2H, dr); ^13C -NMR (100 MHz, DMSO- _d6 ) δ: 30.29, 34.21, 47.23, 54.95 , 68.18, 68.30, 102.42, 115.77, 115.87 (d, J = 21.9 Hz), 127.28, 140.35, 160.60, 161.92 (d, J = 243.2 Hz); IR (KBr): 3300, 3113, 2930, 1638, 1547, 1508, ₁₄₅₄ , 1234, ₈₃₉ cm ^-1 ; MS ( _EI ): m/z 303 (M ⁺ ); HRMS: Calcd for C16H18FN3O2 _303.1383 , Found 303.1388.

Compound 3: 3-benzyl-N-(trans-4-hydroxycyclohexyl)-1H-pyrazole-5-carboxamide
mp: 94-96 °C; ^1H -NMR (400 MHz, _CD3OD ) δ: 1.29-1.46 (4H, m), 1.96-1.98 (4H, m), 3.49-3.61 (1H, m), 3.74- 3.87 (1H, m), 4.02 (2H, s), 6.47 (1H, s), 7.20-7.32 (5H, m); ^13C -NMR (100 MHz, _CD3OD ) δ: 31.44, 32.57, 34.84, 70.35, 105.47, 127.64, 129.56, 129.66, 139.67, 145.39, 148.11, 164.23; m ^/ z 299 (M ⁺ ); _HRMS : Calcd for _{C17H21N3O2 299.1634} , Found _299.1629 .

Compound 4: 3-(4-fluorobenzyl)-N-(trans-4-hydroxycyclohexyl)-1H-pyrazole-5-carboxamide
mp: 95-96 °C; ^1H -NMR (400 MHz, _CD3OD ) δ: 1.34-1.46 (4H, m), 1.96-1.98 (4H, m), 3.47-3.64 (1H, m), 3.73- 3.87 (1H, m), 4.01 (2H, s), 6.47 (1H, s), 7.00-7.04 (2H, m), 7.22-7.25 (2H, m); ^13C -NMR (100 MHz, _CD3OD ) δ: 31.48, 31.64, 34.90, 70.42, 105.40, 116.29 (d, J = 21.9 Hz), 131.34 (d, J = 8.9 Hz), 135.60, 145.49, 148.15, 163.15 (d, J = 242.1 Hz), 164 ,36; IR (KBr): 3300, 3215 _, 2932, 1636, 1541, 1508, 1456, ₁₂₂₃ cm ^-1 ; MS (EI): m/z 317 (M ⁺ ); _{3O2 317.1540} , Found 317.1541.

Compound 5: 3-(4-cyclobenzyl)-N-(trans-4-hydroxycyclohexyl)-1H-pyrazole-5-carboxamide
mp: 88-89 °C; ^1H -NMR (400 MHz, _CD3OD ) δ: 1.34-1.45 (4H, m), 1.96-1.98 (4H, m), 3.49-3.61 (1H, m), 3.73- 3.87 (1H, m), 4.01 (2H, s), 6.48 (1H, s), 7.21 ⁽ 2H, d, J = 8.4 Hz), 7.30 (2H, d, J = 8.4 Hz); 100 MHz, _CD3OD ) delta: 29.56, 31.49, 31.74, 32.14, 34.92, 70.43, 105.50, 129.76, 131.23, 133.57, 138.47, 145.04, 148.26, 164.24; (KBr): 3317, 3300, 3263, 2930, 2864, 1638, 1545, 1491, 1456, ₁₀₈₈ cm ^-1 _; MS (EI): m/z 333 (M ⁺ ₎ ; HRMS: Calcd for _C17H20ClN3O2 333.1244, Found 333.1235.

Compound 6: 3-(2-cyclobenzyl)-N-(trans-4-hydroxycyclohexyl)-1H-pyrazole-5-carboxamide
mp: 96-97°C; ^1H -NMR (400 MHz, _CD3OD ) δ: 1.32-1.43 (4H, m), 1.96-1.98 (4H, m), 3.50-3.61 (1H, m), 3.74- 3.84 (1H, m), 4.15 (2H, s), 6.42 (1H, s), 7.21-7.31 (3H, m), 7.39-7.42 (1H, m); ^13C -NMR (100 MHz, DMSO-d ₆ ) δ: 28.94, 29.65, 30.33, 34.27, 47.04, 68.32, 104.32, 127.57, 128.71, 129.41, 130.86, 132.96, 136.14, 141.78, 147.22, 161.00; IR (KBr): 3375, 3315, 3138, 2934, 2860 , 1638, 1541, 1454 cm ^-1 ; MS (EI): m/z 333 (M ⁺ ) _; HRMS: Calcd for _{C17H20ClN3O2 333.1244} , Found _333.1241 .

Compound 7: 3-(3-cyclobenzyl)-N-(trans-4-hydroxycyclohexyl)-1H-pyrazole-5-carboxamide
mp: 83-85 °C; ^1H -NMR (400 MHz, _CD3OD ) δ: 1.29-1.46 (4H, m), 1.96-1.98 (4H, m), 3.55-3.56 (1H, m), 3.71- 3.88 (1H, m), 4.02 (2H, s), 6.51 (1H, s), 7.15-7.30 (4H, m); ^13C -NMR (100 MHz, _CD3OD ) δ: 29.56, 31.47, 32.58, IR (KBr): 3377, 3254, 3078, 2932 , 2855, 1638, 1545, 1456, 1431, 1078 cm ^{- 1} ; MS (EI) _: m/z 333 (M ⁺ ); _HRMS : Calcd for _{C17H20ClN3O2 333.1244} , Found 333.1249.

Compound 8: N-(trans-4-hydroxycyclohexyl)-3-(4-(trifluoromethyl)benzyl)-1H-pyrazole-5-carboxamide
mp: 94-96 °C; ^1H -NMR (400 MHz, _CD3OD ) δ: 1.34-1.46 (4H, m), 1.96-1.98 (4H, m), 3.47-3.64 (1H, m), 3.73- 3.87 (1H, m), 4.12 (2H, s), 6.51 (1H, s), 7.42 (2H, d, J = 8.0 Hz), 7.61 (2H, d, J = 8.0 Hz); ¹³ C-NMR ( 100 MHz, CD ₃ OD) δ: 31.48, 32.18, 34.91, 70.42, 105.67, 125.76 (q, J = 270.0 Hz), 126.57, 130.28, 144.25, 144.51, 148.35, 164.16; KBr): 3308, 3263, 2937, 1638, 1541, ₁₄₅₆ , 1327, 1124 _, 1067 cm ^-1 ; MS _{( EI} ): m/z 367 (M ⁺ ); HRMS: _Calcd for C18H20F3N3O2 367.1508, Found 367.1507.

Compound 9: N-(trans-4-hydroxycyclohexyl)-3-(3-methoxybenzyl)-1H-pyrazole-5-carboxamide
mp: 65-67 °C; ^1H -NMR (400 MHz, _CD3OD ) δ: 1.29-1.46 (4H, m), 1.96-1.98 (4H, m), 3.55-3.58 (1H, m), 3.76- 3.80 (4H, m), 3.99 (2H, s), 6.48 (1H, s), 6.79-6.81 (3H, m), 7.20 (2H, t, J = 8.0 Hz); ^13C -NMR (100 MHz, _CD3OD ) δ: 31.47, 32.47, 34.89, 55.61, 70.40, 105.46, 113.07, 115.31, 121.86, 130.69, 141.69, 145.51, 148.16, 161.41, 164.2 8; IR (KBr): 3373, 3360, 3283, 2999, 2901 , 2858, 1612, 1508, 1491, 1456, 1508 cm ^-1 ; MS (EI): m/z 329 (M ⁺ ); HRMS: Calcd for C ₁₈ H ₂₃ N ₃ O ₃ 329.1739, Found 329.1743.

Compound 10: N-(trans-4-hydroxycyclohexyl)-3-(4-methoxybenzyl)-1H-pyrazole-5-carboxamide
mp: 75-77 °C; ^1H -NMR (400 MHz, _CD3OD ) δ: 1.33-1.41 (4H, m), 1.96-1.98 (4H, m), 3.50-3.61 (1H, m), 3.76- 3.82 (4H, m), 3.95 (2H, s), 6.45 (1H, s), 6.86 (2H, d, ^J = 8.6 Hz), 7.14 (2H, d, J = 8.6 Hz); 100 MHz, _CD3OD ) δ: 31.49, 31.63, 34.90, 55.70, 70.42, 105.30, 115.09, 130.59, 131.55, 146.10, 148.10, 159.98, 164.32; IR (KBr): 3 261, 2930, 1647, 1541, 1508, 1456, 1246 _cm ^-1 ; MS (EI): m/z 329 (M ⁺ ) _; HRMS: Calcd for _{C18H23N3O3 329.1739} , Found 329.1746.

Compound 11: N-(trans-4-hydroxycyclohexyl)-3-(2-methoxybenzyl)-1H-pyrazole-5-carboxamide
mp: 71-72 °C; ^1H -NMR (400 MHz, _CDCl3 ) δ: 1.24-1.49 (4H, m), 1.98-2.08 (4H, m), 3.59-3.66 (1H, m), 3.86-3.94 (4H, m), 3.97 (2H, s), 6.61 (1H, s), 6.90-6.93 (2H, m), 7.15 (1H, dd, J = 8.0, 2.4 Hz), 7.23-7.28 (1H, m ]; ^30.13 , 144.74 _, 146.62, 156.87, 161.58 IR (KBr): 3412, 3337, 3227, 1641, 1551, 1495, 1456, 1246 cm ^-1 _; MS (EI): m/z 329 (M ⁺ ); HRMS: _Calcd for _{C18H23N3O 3} 329.1739, Found 329.1738.

Compound 12: N-(trans-4-hydroxycyclohexyl)-3-(4-methylbenzyl)-1H-pyrazole-5-carboxamide
mp: 86-87 °C; ^1H -NMR (400 MHz, _CD3OD ) δ: 1.34-1.46 (4H, m), 1.96-1.98 (4H, m), 2.29 (3H, s), 3.47-3.63 ( 1H, m), 3.72-3.86 (1H, m), 3.96 (2H, s), 6.45 (1H, s), 7.08-7.14 (4H, m); ^13C -NMR (100 MHz, _CD3OD ) δ IR (KBr): 3327, 3 234, 3177, 2925, 2864, 1645, 1543, 1516, 1456, 1082 _cm ^-1 _; MS (EI): m/z 313 (M ⁺ ); HRMS: Calcd for _{C18H23N3O2 313.1790} , Found 313.1790.

Compound 13: N-(trans-4-hydroxycyclohexyl)-3-phenethyl-1H-pyrazole-5-carboxamide
mp: 91-93°C; ^1H -NMR (400 MHz, _CD3OD ) δ: 1.34-1.47 (4H, m), 1.97-1.99 (4H, m), 2.89-3.02 (4H, m), 3.55- 3.57 (1H, m), 3.75-3.88 (1H, m), 6.48 (1H, m), 7.14-7.18 (3H, m), 7.25 (2H, t, J = 3.6 Hz); ¹³ C-NMR (100 MHz, _CD3OD ) δ: 28.36, 31.51, 34.88, 36.49, 49.16, 70.39, 104.85, 127.23, 129.42, 129.45, 142.02, 146.06, 148.01, 164.45; IR (KBr) : 3368, 3179, 2930, 2864, 1649 , 1543, 1456, 1086, 700 cm ^-1 ; MS (EI): m/z 313 (M ⁺ ); HRMS: Calcd for C ₁₈ H ₂₃ N ₃ O ₂ 313.1790, Found 313.1781.

Compound 14: 3-benzyl-N-(4-oxocyclohexyl)-1H-pyrazole-5-carboxamide
mp: 131-132 °C; ^1H -NMR (400 MHz, _CDCl3 ) δ: 1.73-1.84 (2H, m), 2.27-2.31 (2H, m), 2.40-2.53 (4H, m), 4.04 (2H , s), 4.35-4.44 (1H, m), 6.62 (1H, s), 6.80 (1H, d, J = 6.8 Hz), 7.22 (2H, d, J = 6.8 Hz), 7.26 (1H, tt, J = 6.8, 1.6 Hz), 7.32 (2H, tt, J = 6.8, _1.6 ^Hz ); , 128.81, 137.44, 144.92, 146.26, 161.70, 210.11; IR (KBr): 3300, 3192, 1715, 1638, 1545, 1456 cm ^-1 ; MS (EI): m/z 297 (M ⁺ ); for _{C17H19N3O2 297.1477} , Found _{297.1486 .}

Compound 17: 3-(4-hydroxyphenyl)-N-methyl-1H-pyrazole-5-carboxamide
mp: 177-179 °C; ^1H -NMR (400 MHz, DMSO- _d6 ) δ: 2.74 (3H, d, J = 4.0 Hz), 6.82 (2H, d, J = 8.4 Hz), 6.89 (1H, s), 7.58 (2H, d, J = 8.4 Hz), 8.18 ( ^1H , d, J = _3.6 Hz); 126.87, 145.29, 145.73, 157.85, 161.54; IR (KBr): 3219, 3107, 3042, 1655, 1614, 1508 cm ^-1 ; MS (EI): m/z 217 (M ⁺ ); HRMS: Calcd for _{C11 H11N3O2 217.0851} , Found 217.0841 _.

Compound 18: 3-benzyl-N-methyl-1N-pyrazole-5-carboxamide
mp: 168-170 °C; ^1H -NMR (400 MHz, _CDCl3 ) δ: 2.95 (3H, d, J = 4.8 Hz), 4.03 (2H, s), 6.58 (1H, s), 6.80 (1H, s), 7.19-7.34 (5H, m); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ: 25.56, 31.00, 103.95, 126.48, 128.49, 128.56, 138.80, 143.42, 147.18, 162.46; IR (KBr ): 3084, 1638, 1570 cm ^-1 ; _MS (EI): m/z 215 (M ⁺ ); HRMS: Calcd for _{C12H13N3O 215.1059} , Found 215.1055.

Compound 19: 3-benzyl-N,N-dimethyl-1N,N-dimethyl-1H-pyrazole-5-carboxamide
mp: 149-151 °C; ^1H -NMR (400 MHz, _CDCl3 ) δ: 3.10 (3H, s), 3.27 (3H, s), 4.03 (2H, s), 6.35 (1H, s), 7.21- 7.24 (3H, m), 7.31 (2H, t, J = 7.4 Hz); ¹³ C-NMR (100 MHz, _CDCl3 ) δ: 33.43, 36.28, 38.79, 106.46, 126.51, 128.59, 128.71, 138.69, 141.21 , 148.27, _162.54 ; IR (KBr): 3128, 2968, 2860, 1599, 1495 _, 1398, 719 cm ^-1 ; MS (EI): m/z 229 (M ⁺ ); HRMS: Calcd for _C13H15N3 O 229.1215, Found 229.1219.

Compound 20: 3-benzyl-N-methoxy-N-methyl-1H-pyrazole-5-carboxamide
¹ H-NMR (400 MHz, CDCl ₃ ) δ: 3.36 (3H, s), 3.73 (3H, s), 4.07 (2H, s), 6,57 (1H, s), 7.20-7.32 (5H, m ); ¹³ C-NMR (100 MHz, CDCl ₃ ) δ: 32.93, 34.27, 61.46, 107.88, 126.30, 128.47, 128.70, 135.97, 139.33, 152.05, 159.72; IR (neat): 3213, 3155, 1626, 1558, 1495, 1456, 1435 cm ^-1 ; MS (EI ₎ : m/z 245 (M ⁺ ); HRMS: Calcd for _{C13H15N3O2 245.1164} , Found _245.1161 .

Compound 21: (3-benzyl-1H-pyrazol-5-yl)(piperidin-1-yl)methanone
mp: 151-153 °C; ^1H -NMR (400 MHz, _CDCl3 ) δ: 1.61-1.70 (6H, m), 3.59-3.83 (4H, m), 4.03 (2H, s), 6.31 (1H, s) ), 7.22-7.26 ( ^4H , m), 7.29-7.33 (2H _, m); , 128.67, 128.76, 138.47, 142.18, 147.64, 161.49; IR (KBr): 3146, 2924, 1593, 1506, 1421, 1250, 725 cm ^-1 ; MS (EI): m/z 269 (M ⁺ ); : Calcd _{for C16H19N3O 269.1528} , Found 269.1537.

Compound 22: Azepan-1-yl (3-benzyl-1H-pyrazol-5-yl)methanone
mp: 145-147 °C; ^1H -NMR (400 MHz, _CDCl3 ) δ: 1.58-1.59 (4H, m), 1.77-1.80 (4H, m), 3.65 (2H, t, J= 6.0 Hz), 3.72 (2H, t, J = 6.0 Hz), 4.05 (2H, s), 6.31 (1H, s), 7.22-7.26 (3H, m), 7.31 (2H, t, J = 7.2 Hz); ^13C- NMR (100 MHz, _CDCl3 ) delta: 26.88, 27.27, 27.52, 29.42, 33.57, 46.72, 48.68, 106.00, 126.48, 128.58, 128.71, 138.77, 141.09, 148.80, 162.13; IR (KBr): 3171, 3128, 3086 , 2978, 2924, 2853, 1576, 1510, 1454, 1421, 719 cm ^-1 ; MS (EI): m/z 283 (M ⁺ ); HRMS: Calculated for C ₁₇ H ₂₁ N ₃ O 283.1685, Found 283.1675.

Compound 23: (3-benzyl-1H-pyrazol-5-yl)(morpholino)methanone
mp: 122-124 °C; ^1H -NMR (400 MHz, _CDCl3 ) δ: 3.72-3.91 (8H, m), 4.05 (2H, s), 6.41 (1H, s), 7.22-7.35 (5H, m ); ¹³ C-NMR (100 MHz, CDCl ₃ ) δ: 32.61, 42.81, 47.47, 66.90, 106.70, 126.78, 128.766, 128.73, 137.89, 143.80, 145.76, 162.22; IR (KBr) : 3119, 2966, 2853, 1585, 1499, 1238, 1109 cm ^-1 ; MS ( _EI ): m/z 271 (M ⁺ ) _; HRMS: Calcd for _{C15H17N3O2 271.1321} , Found 271.1322.

Compound 24: (3-benzyl-1H-pyrazol-5-yl)(4-hydroxypiperidin-1-yl)methanone
¹ H-NMR (400 MHz, CD ₃ OD) δ: 1.44-1.59 (2H, m), 1.81-1.97 (2H, m), 3.30-3.48 (2H, m), 3.85-3.91 (1H, m), 4.03 (2H, s), 4.09-4.31 (2H, m), 6.32 (1H, s), 7.20-7.32 (5H, m); ^13C -NMR (100 MHz, _CD3 OD) δ: 32.40, 34.83, IR (neat): 3676, 3387, 3327, 3294, 3179, 2955, 1611, 1597, 1506, 1495 cm ^{- 1} ; MS (EI): m/z 285 (M ⁺ ); _HRMS : Calcd for _{C16H19N3O2 285.1477} , Found 285.1468 _.

(4) Synthesis of compound 15

Selenium dioxide (420 mg, 3.78 mmol) was added to a mixed solution of acetophenone 4 (300 mg, 2.50 mmol) in 1,4-dioxane and water (15 mL, 3:1) under Ar atmosphere at room temperature. After that, the mixture was stirred under heating under reflux for 18 hours. After allowing to cool, the reaction solution was filtered, and the solvent was distilled off using a rotary evaporator. Water (5 mL) was added to the obtained residue, and the mixture was further stirred under reflux with heating for 5 hours. After completion of the reaction, the solvent was distilled off using a rotary evaporator, and the resulting residue was purified by silica gel column chromatography (n-hexane:ethyl acetate = 1:1) to give gem-diol 5 (85 mg, 22 %, 0.55 mmol).
p-Toluenesulfonyl hydrazide (114 mg, 0.61 mmol) and ethyl acrylate (0.08 mL, 0.77 mmol) were added to a solution of gem-diol 5 (78 mg, 0.51 mmol) in dimethyl sulfoxide (2.5 mL) under Ar atmosphere. , and cesium carbonate (500 mg, 1.54 mmol) were sequentially added, followed by stirring at 100° C. for 5 hours. After completion of the reaction, water (3 mL) was added and further diluted with ethyl acetate (5 mL). After separating the organic layer, the aqueous layer was extracted with ethyl acetate (1.5 mL × 3) and combined with the previous organic layer. After drying the organic layer with anhydrous sodium sulfate, the solvent was distilled off using a rotary evaporator, and the resulting residue was purified by silica gel column chromatography (n-hexane:ethyl acetate = 5:1) to give pyrazole. Ester 6 (27 mg, 22%, 0.11 mmol) was obtained.

Under Ar atmosphere, to a solution of pyrazole ester 6 (27 mg, 0.12 mmol) in ethanol (3 mL), 10% NaOH aqueous solution (1.5 mL) was added at room temperature, and the mixture was stirred under reflux with heating for 2 hours. After completion of the reaction, ethanol was distilled off, 10% HCl aqueous solution (3 mL) was added, and the aqueous layer was extracted with ethyl acetate (1.5 mL x 3). After the organic layer was dried using anhydrous sodium sulfate, the solvent was distilled off using a rotary evaporator, and the resulting carboxylic acid was used in the next reaction without purification. Under Ar atmosphere, the corresponding trans-4-aminocyclohexanol (22 mg, 0.19 mmol), triethylamine (0.03 mL, 0.75 mmol), 1 -Hydroxybenzotriazole (25 mg, 0.75 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (36 mg, 0.75 mmol) were sequentially added, and the mixture was stirred at room temperature for 20 hours. After completion of the reaction, the solvent was distilled off using a rotary evaporator, and the resulting residue was purified by silica gel column chromatography (n-hexane: acetone = 1: 3) to give the desired compound 15 (17 mg, 44 % in 2 steps, 0.06 mmol).

Compound 15: 3-benzoyl-N-(trans-4-hydroxycyclohexyl)-1H-pyrazole-5-carboxamide
mp: 83-84 °C; ^1H -NMR (400 MHz, _CDCl3 ) δ: 1.31-1.53 (4H, m), 2.03-2.14 (4H, m), 3.64-3.71 (1H, m), 3.93-4.02 (1H, m), 6.76 (1H, d, J = 7.6 Hz), 7.36 (1H, s), 7.54 (2H, t, J = 7.6 Hz), 7.66 (1H, t, J = 7.6 Hz), 8.01 (2H, d, J = 7.6 Hz); ¹³ C-NMR (100 MHz, acetone-d ₆ ) δ: 31.34, 48.75, 49.75, 55.47, 69.78, 109.57, 128.98, 129.32, 130.57, 131.25, 133.96 , 137.96, 159.74, 186.18; IR ( _KBr ): 3433, 3344, 2856, 1647, 1545, ₁₃₁₉ cm ^-1 ; MS ( _EI ): m/z 313 (M ⁺ ); HRMS: Calcd for _C17H19N3O3 313.1426, Found 313.1423.

(5) Synthesis of compound 16

Under Ar atmosphere, hexamethylenetetramine (845 mg, 6.03 mmol) was added to a solution of 2-bromo-1-phenylethanone 7 (1 g, 5.02 mmol) in chloroform (15 mL) at room temperature. Stirred at 60° C. for 6 hours. After allowing to cool, the precipitated solid was collected by filtration, and the obtained solid was dissolved in a mixed solution of ethanol and concentrated hydrochloric acid (20 mL, 3:1), and the mixture was further stirred under reflux with heating for 16 hours. After completion of the reaction, the solvent was distilled off using a rotary evaporator, and the resulting hydrochloride was used in the next reaction without purification. Ethyl thiooxamate (669 mg, 5.02 mmol) and sodium acetate (989 mg, 12.06 mmol) were sequentially added to a solution of the obtained hydrochloride in acetic acid (15 mL) at room temperature under an Ar atmosphere, followed by heating. The mixture was stirred under reflux for 19 hours. After completion of the reaction, the reaction solution was diluted with ethyl acetate (20 mL), and saturated aqueous sodium bicarbonate solution (20 mL) was added. After separating the organic layer, the aqueous layer was extracted with ethyl acetate (5 mL × 3) and combined with the previous organic layer. After drying the organic layer with anhydrous sodium sulfate, the solvent was distilled off using a rotary evaporator. was obtained imidazole ester 8 (608 mg, 56% in 3 steps, 2.81 mmol).

Under an Ar atmosphere, to a solution of imidazole ester 8 (220 mg, 1.02 mmol) in ethanol (5 mL), 10% NaOH aqueous solution (2.5 mL) was added at room temperature, and the mixture was stirred under reflux for 2 hours. After completion of the reaction, ethanol was distilled off, 10% HCl aqueous solution (5 mL) was added, and the aqueous layer was extracted with ethyl acetate (1.5 mL x 3). After the organic layer was dried using anhydrous sodium sulfate, the solvent was distilled off using a rotary evaporator, and the resulting carboxylic acid was used in the next reaction without purification. Under Ar atmosphere, the corresponding trans-4-aminocyclohexanol (175 mg, 1.52 mmol), triethylamine (0.21 mL, 1.52 mmol), 1 -Hydroxybenzotriazole (206 mg, 1.52 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (292 mg, 1.52 mmol) were sequentially added, and the mixture was stirred at room temperature for 20 hours. After completion of the reaction, the solvent was distilled off using a rotary evaporator, and the resulting residue was purified by silica gel column chromatography (n-hexane: acetone = 1: 3) to give the desired compound 16 (171 mg, 59 % in 2 steps, 0.60 mmol).

Compound 17: N-(trans-4-hydroxycyclohexyl)-5-phenyl-1H-imidazole-2-carboxamide
mp: 178-180 °C; ^1H -NMR (400 MHz, _CD3OD ) δ: 1.37-1.53 (4H, m), 1.99-2.05 (4H, m), 3.57-3.63 (1H, m), 3.80- 3.86 (1H, m), 7.27 (1H, t, J = 7.2 Hz), 7.39 (2H, t, J = 7.2 Hz), 7.57 (1H, s), 7.76 (1H, d, J = 7.2 Hz); ¹³ C-NMR (100 MHz, CD ₃ OD) δ: 31.43, 34.75, 49.30, 70.28, 111.49, 118.65, 126.20, 127.16, 128.24, 128.50, 129.82, 133.98, 142.52, 159.58; IR (KBr): 3311, 3281 , 3204, 1653, 1634, 1549, 1456 cm ^-1 ; MS (EI): m/z 285 (M ⁺ ); HRMS: Calcd for C ₁₇ H ₂₀ N ₂ O ₂ 285.1477, Found 285.1484.

(6) Synthesis of Comparative Compounds 1 and 2

Synthesis of comparative compound 1
Di-tert-butyl dicarbonate (121 mg, 0.56 mmol) and N,N-dimethylaminopyridine (10 mg, 0.09 mmol) were sequentially added, and the mixture was stirred at room temperature for 23 hours. After completion of the reaction, saturated aqueous sodium bicarbonate (3 mL) was added. After separating the organic layer, the aqueous layer was extracted with dichloromethane (1.5 mL × 3) and combined with the previous organic layer. After drying the organic layer with anhydrous sodium sulfate, the solvent was distilled off using a rotary evaporator. A Boc protected form (111 mg, 82%, 0.35 mmol) was obtained.
Phenylboronic acid (60 mg , 0.50 mmol) and tetrakis(triphenylphosphine)palladium (12 mg, 0.01 mmol) were sequentially added, and the mixture was stirred at 110° C. for 40 hours. After completion of the reaction, the solvent was distilled off using a rotary evaporator, and the resulting residue was purified by silica gel column chromatography (n-hexane:ethyl acetate = 2:1) to give the desired pyrrole ester 10 (26 mg , 62%, 0.12 mmol).

Under an Ar atmosphere, a 10% NaOH aqueous solution (0.5 mL) was added to a solution of pyrrole ester 10 (34 mg, 0.16 mmol) in ethanol (1 mL) at room temperature, and the mixture was stirred under reflux for 2 hours. After completion of the reaction, ethanol was distilled off, 10% HCl aqueous solution (1 mL) was added, and the aqueous layer was extracted with ethyl acetate (1 mL x 5). After the organic layer was dried using anhydrous sodium sulfate, the solvent was distilled off using a rotary evaporator, and the resulting carboxylic acid was used in the next reaction without purification. Under Ar atmosphere, the corresponding trans-4-aminocyclohexanol (28 mg, 0.24 mmol), triethylamine (0.03 mL, 0.24 mmol), 1 -Hydroxybenzotriazole (32 mg, 0.24 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (46 mg, 0.24 mmol) were sequentially added, and the mixture was stirred at room temperature for 20 hours. After completion of the reaction, the solvent was distilled off using a rotary evaporator, and the resulting residue was purified by silica gel column chromatography (n-hexane: acetone = 1: 3) to give the desired comparative compound 1 (27 mg, 58% in 2 steps, 0.10 mmol).

Comparative compound 1: N-(trans-4-hydroxycyclohexyl)-4-phenyl-1H-pyrrole-2-carboxamide
mp: 241-243 °C; ^1H -NMR (400 MHz, _CD3OD ) δ: 1.36-1.50 (4H, m), 1.97-2.01 (4H, m), 3.54-3.59 (1H, m), 3.81- 3.82 (1H, m), 7.12-7.16 (2H, m), 7.25 (1H, d, J = 1.2 Hz), 7.30 (2H, t, J = 8.0 Hz), 7.53 (2H, dd, J = 8.0, 1.2 Hz); ¹³ C-NMR (100 MHz, CD ₃ OD) δ: 31.78, 35.11, 49.17, 70.60, 108.94, 119.59, 125.95, 126.71, 127.05, 127.95, 129.68, 136.79, 1 62.95; IR (KBr): 3439 , 3302, 2936, 1622, 1570, 1541, 756 cm ^-1 ; MS (EI): m/z 284 (M ⁺ ); HRMS: Calcd for C ₁₇ H ₂₀ N ₂ O ₂ 284.1525.

Synthesis of comparative compound 2
To a mixed solution of ethyl 5-bromo-1H-pyrrole-2-carboxylate 11 (95 mg, 0.44 mmol) in toluene and saturated aqueous sodium carbonate (2 mL, 3:1) under Ar atmosphere at room temperature After sequentially adding phenylboronic acid (80 mg, 0.65 mmol) and tetrakis(triphenylphosphine)palladium (25 mg, 0.02 mmol), the mixture was stirred at 100° C. for 20 hours. After completion of the reaction, the solvent was distilled off using a rotary evaporator, and the resulting residue was purified by silica gel column chromatography (n-hexane:ethyl acetate = 2:1) to give the desired pyrrole ester 12 (40 mg , 43%, 0.19 mmol).
Under an Ar atmosphere, to a solution of pyrrole ester 12 (37 mg, 0.17 mmol) in ethanol (1 mL), 10% NaOH aqueous solution (0.5 mL) was added at room temperature, and the mixture was stirred under reflux for 2 hours. After completion of the reaction, ethanol was distilled off, 10% HCl aqueous solution (1 mL) was added, and the aqueous layer was extracted with ethyl acetate (1 mL x 5). After the organic layer was dried using anhydrous sodium sulfate, the solvent was distilled off using a rotary evaporator, and the resulting carboxylic acid was used in the next reaction without purification. Under Ar atmosphere, the corresponding trans-4-aminocyclohexanol (31 mg, 0.27 mmol), triethylamine (0.04 mL, 0.27 mmol), 1 -Hydroxybenzotriazole (37 mg, 0.27 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (52 mg, 0.27 mmol) were sequentially added, and the mixture was stirred at room temperature for 20 hours. After completion of the reaction, the solvent was distilled off using a rotary evaporator, and the resulting residue was purified by silica gel column chromatography (n-hexane: acetone = 1: 3) to give the desired comparative compound 2 (32 mg, 62% in 2 steps, 0.11 mmol).

Comparative compound 2: N-(trans-4-hydroxycyclohexyl)-5-phenyl-1H-pyrrole-2-carboxamide
mp: 232-234 °C; ^1H -NMR (400 MHz, _CD3OD ) δ: 1.35-1.49 (4H, m), 1.99-2.01 (4H, m), 3.55-2.57 (1H, m), 3.75- 3.90 (1H, m), 6.52 (1H, d, J = 4.0 Hz), 6.87 (1H, d, J = 4.0 Hz), 7.24 (1H, t, J = 7.6 Hz), 7.38 (1H, t, J = 7.6. Hz), 7.66 (2H, ^dd , J = 7.6, _1.2 Hz); 129.90, 133.44, 137.06, 162.95; IR (KBr): 3414, 3319, 3227, 1624, 1541, 1458, 1339, 1273, 756 cm ^-1 ; MS (EI): m/z 284 (M ⁺ ); HRMS: Calcd _{for C17H20N2O2 284.1525} , Found _284.1521 .

Structural formulas of the compounds synthesized by the above method are shown in FIGS. 1-1, 1-2 and 1-3.

Experimental example
1. Experiment material (1) Reagent
6-diazo-5-oxo-L-norleucine (DON) (Wako Pure Chemical), CB-839 (Selleck), 3-Phenyl-1H-1,2,4-triazol-5-amine(1) (Vitas M Chemical, Hong Kong), Compound (2) (7127371, Otava Chemicals, Vilniaus, LITHUANIA), Compound (3) (P2001S-209195, Pharmeks, Moscow, Russia), N-(Pyridin-2-ylmethyl)-5-( thiophen-2-yl)-1H-pyrazole-3-carbboxamide(4) (F5791-2259, Life Chemicals, Ontario, Canada), 5-(Furan-2-yl)-N-[(pyridin-3-yl) methyl]-1H-pyrazole-3-carboxamide(5) (Z317489074, Enamine, Kiev, Ukraine), 5-(Furan-2-yl)-N-(pyridin-2-ylmethyl)-1H-pyrazole-3-carboxamide (6) (Z317488928, Enamine, Kiev, Ukraine), N-((3-(2-Oxo-1,2-dihydropyridin-3-yl)-1,2,4-oxadiazol-5-yl)methyl)- 1H-pyrazole-5-carboxamide (7) (Life Chemicals), 2-hydroxy-4-oxo-N-(pyridin-2-ylmethyl)-4H-pyrido[1,2-a]pyrimidine-3-carboxamide (8 ) (ChemDiv) were used after being dissolved in DMSO.

(2) Experimental animals Mice (C57BL/6JJcl, male, 8 weeks old, Sankyo Labo Service Co., Ltd.) were fed standard chow (CLEA Rodent Diet CE-2 for mice, rats and hamsters, CLEA Japan, Inc.) and tap water. were given free drinking water and reared in a laboratory animal room controlled at constant temperature and constant humidity (24°C, 30%) with a 12-hour light-dark cycle (8:00 to 20:00). This experiment was conducted in full consideration of animal ethics after being approved by the Animal Experimentation Committee based on the National University Corporation Toyama University Animal Experiment Handling Regulations.

2. Construction of mouse recombinant GLS1 Primers G1U1 (5'- cccatatgatgcggctgcgaggctcggcg-3' (SEQ ID NO: 1), NdeI site) and G1-204L1 (5'- ggggatccttatagcaacccgtcgagattcttg-3' (SEQ ID NO: 2), BamHI site), KOD-Neo DNA A 2.0 kb DNA fragment of mouse GLS1 was amplified from a mouse brain cDNA library using polymerase (TOYOBO). The amplified DNA fragment was subcloned into the EcoRV site of pLITMUS28 (NEB) to obtain plasmid pLITG1. After confirming the DNA base sequence with a Genetic Analyzer (ABI), the NdeI-BamHI DNA fragment of pLITG1 was cloned into the NdeI and BamHI sites of the pOPHLT expression vector to obtain pOPG1. pOPG1 was transformed into Origami B (DE3) competent cells (Novagen).
5 μl of glycerol stock (pO-Gls1-204 in Origami), 5 ml of LB and 5 μl of ampicillin were mixed and shaken overnight at 37°C.

500 ml of LB medium was prepared by adding 10 g of Bacto Peptone (Thermo), 5 g of DRIED YEAST EXTRACT (Wako Pure Chemical), and 5 g of sodium chloride to 1 L of deionized water and sterilizing. °C until OD600 nm = 0.6. 2.5 ml of 100 mM IPTG (isopropyl-β-D(-)-thiogalactopyranoside) (TaKaRa) was added and shaken at 25°C for 16 hours or more to induce the expression of the His-tagged recombinant protein. After centrifugation at 5000 rpm, 15 min, 4° C., the supernatant was discarded and suspended in 30 ml of 50 mM Tris-HCl (pH 8.0). After centrifuging at 10,000 rpm, 5 min, 25°C and discarding the supernatant, the pellet was stored at -80°C. The pellet was subjected to Ni column purification using a HiTagFF column (Pharemacia), subjected to 10% SDS-PAGE, and stained with Quick CBB Plus (Wako Pure Chemical Industries) to determine fractions containing the target protein. A fraction containing the target protein was placed in a dialysis tube (Spectra/Por, REPLIGEN) and dialyzed overnight at 4°C in a dialysis buffer (50 mM Tris-phosphate pH 8.0, 1 mM DTT, 10% glycerol). A solution containing GLS1 was concentrated to about 1 mg/ml using a centri-tube, and after confirming the enzymatic activity, it was used in the experiment.

3. Generation of human recombinant GLS1
DNA encoding the hGLS1 amino acid sequence (residues 73-669) was inserted into the NdeI and BamHI sites of the pOPTH plasmid (Obita et al, 2007), and the resulting plasmid was used to transform E. coli (SoruBL21 strain). did. When the OD600nm of the culture reached 0.5, IPTG was added to a final concentration of 0.5 mM to induce the expression of hGLS1. After overnight culture at 37°C, cells were harvested by centrifugation at 4°C and placed in buffer containing 20 mM Tris-HCl (pH 8.0 at 4°C), 500 mM NaCl, 10% glycerol, 5 mM β-mercaptoethanol. was resuspended in After lysing the cells by applying ultrasonic waves on ice, the cells were centrifuged at 10,200×g for 60 minutes at 4° C. to obtain a supernatant. This His-tagged hGLS1 protein was purified on a Ni-NTA column and then subjected to gel filtration chromatography using a Superdex HiLoad 16/60 column equilibrated with a buffer containing 20 mM Tris (pH 8.0, 4°C) and 100 mM NaCl. Graphically refined. The purified protein consists of hGLS1 (residues 73-669) and an N-terminal His-tag.

4. Evaluation of glutaminase 1 inhibitory activity (1) Evaluation of glutamate production inhibitory activity using mouse kidney lysate Approximately 50 mg of mouse kidney was biomashed with a homogenate buffer (0.25 M Sucrose, 5 mM Tris, 1 mM EDTA/2Na, pH 7.4). III (Nippi Co., Ltd.) and centrifuged at 4° C., 6000 g for 30 seconds to obtain a supernatant. Absorbance was then measured with an iMark microplate reader (BIO-RAD, 595 nm) by the Bradford method using Quick Start protein assay (5000201JA, BIO-RAD). Homogenate sample (5 mg/mL), GLS1 inhibitor candidate compound (test compound) dissolved in solvent (DMSO) or solvent alone (for control), reaction buffer (27.5 mM Tris, 0.11 mM EDTA 2Na, 110 mM KH _{2 PO4} , pH 8.6) and incubated at 37°C for 20 minutes. After adding an L-glutamine solution (40 mM) and incubating at 37° C. for 10 minutes, 10 μL of 3N HCl was added to terminate the reaction. Glu concentration was determined by measuring absorbance at 560 nm with a multifunction plate reader (GENios, Tecan) using L-glutamic acid measurement kit "Yamasa" NEO (Yamasa Soy Sauce Co., Ltd.). The Glu concentration was calculated from the following formula according to the instructions of the kit.

L-glutamic acid concentration (mg/L)
= (Sample measurement value - Blank)) / (STD measurement value - Blank) x 250

(2) Evaluation of Glu production inhibitory action using mouse and human recombinant GLS1 Reaction solution (Tris-acetate (50 mM, pH 8.6), KPO ₄ (150 mM, pH 8.0), EDTA (0.2 mM, pH 8.0)) 10.5 μL, GLS1 inhibitor candidate compound (test compound) dissolved in solvent (DMSO) or 2.5 μL of solvent alone (for control), and 32 μL of recombinant Gls1 stock were incubated at 37° C. for 20 minutes. L-glutamine solution (20 mM) was added to each reaction and incubated at 37°C for 1 hour. The Glu concentration was determined by measuring absorbance at 555 nm with an ultraviolet/visible spectrophotometer (GeneQuant 1300, Biochrom) using L-glutamic acid measurement kit "Yamasa" NEO.

5. Statistical analysis Measurement data were expressed as mean±standard error. Tukey-Kramer HSD test was performed using JMP Pro version 15.2.0 (SAS Institute Inc, NC, USA) for statistical analysis of the experiment. A p < 0.05 was considered statistically significant.

Experimental Example 1: GLS1 inhibitory activity of compounds selected by in silico screening Primary screening using a three-dimensional pharmacophore obtained from the complex crystal structure of glutaminase and a known GLS1 inhibitor, filtering by Lipinski's "Rule of Five", By performing secondary screening using molecular docking calculations, filtering by binding free energy calculations, filtering based on docking poses, and molecular similarity analysis based on chemical structures, 8 GLS1 inhibitor candidate compounds were identified from a commercial compound database. One compound (Fig. 2(A)) was selected.

For these candidate compounds, the production rate of glutamic acid (Glu) was determined using an evaluation method using mouse kidney lysate, and the GLS1 inhibitory activity was evaluated. As positive control compounds, DON (6-Diazo-5-oxo-L-norleucine), which is a known GLS1 inhibitor shown in the following formula, and CB-839 were used.

DON and CB-839 were dissolved in a solvent (DMSO) to a final concentration of 1 mM. Eight candidate compounds were dissolved in DMSO to a final concentration of 10 mM. As a control, the Glu production rate was also measured in a system in which only the solvent was added instead of the compound.

The results are shown in FIG. 2(B). As shown in the figure, the GLS1 inhibitory activity of candidate compound 3 was the strongest, and the amount of Glu produced was reduced by 53% compared to the control. From the correlation between the structures of candidate compounds 1 to 8 and their GLS1 inhibitory activity, it was speculated that the pyrazole ring and the carbonyl group at the amide site in the structure of candidate compound 3 are important for GLS1 inhibitory activity.

Experimental Example 2: Structure-Activity Relationship In Experimental Example 1, candidate compound 3 exhibited about 50% GLS1 inhibitory activity, and multiple compounds were synthesized using candidate compound 3 as a lead compound. Then, these compounds were evaluated for GLS1 inhibitory activity by determining the production rate of glutamic acid (Glu) using an evaluation method using mouse kidney lysate.

Using the GLS1 inhibitory activity as an index, the structure-activity relationship with the lead compound (candidate compound 3) was examined from the following four points.
1) Modification of the nitrogen substituent of the amide site
2) Conversion of substituents on the benzene ring
3) Increase or decrease the carbon number of the linker between the benzene ring and the pyrazole ring
4) conversion of the pyrazole ring to other heterocycles.

Table 1 shows the correspondence between the symbols of the compounds used in the experiment and the compound numbers described above.

(1) Modification of the nitrogen substituent of the amide moiety , according to the preparation examples described above, the condensation reaction of the known compound (1) with a carboxylic acid and an amine yields a secondary or tertiary amino moiety or a cyclic amine. were synthesized (Scheme 1) and evaluated for their GLS1 inhibitory activity (FIGS. 3(A) and (B)).

その結果、上記式中、RがNHPh基であるアミド誘導体2c以外は、GLS1阻害活性が認められた。但し、Rがピロリジニル基であるアミド誘導体2fのGLS1阻害活性は低かった。これらのアミド誘導体のなかでも、R基がトランス－４－ヒドロキシシクロヘキシルアミノ基であるアミド誘導体2jのGLS1阻害活性が最も強く、その10ｍM濃度でのGlu産生率は、コントロールに比べて99％も低かった（図３（Ｂ））。

As a result, GLS1 inhibitory activity was confirmed except for the amide derivative 2c in which R is an NHPh group in the above formula. However, the GLS1 inhibitory activity of the amide derivative 2f in which R is a pyrrolidinyl group was low. Among these amide derivatives, the amide derivative 2j, in which the R group is a trans-4-hydroxycyclohexylamino group, had the strongest GLS1 inhibitory activity, and its Glu production rate at a concentration of 10 mM was 99% lower than that of the control. (Fig. 3(B)).

(2) Transformation of Benzene Ring Substituents Next, the structure of the amide moiety was fixed, the hydrogen atoms of the benzene ring were replaced with other atoms or functional groups (R), and the GLS1 inhibitory activity was evaluated (Fig. 3 (A ) and (B)). Specifically, commercially available phenylacetones 3a-3j were reacted in two steps to convert them to ethyl esters 4a-4j (Scheme 2), as described in the preparations above. The ester moieties of the ethyl esters 4a-4j were then hydrolyzed, and the resulting carboxylic acids were subjected to a condensation reaction to give derivatives 5a-5j. In addition, deprotection of the methoxymethyl group of derivative 5j gave derivative 5k having an OH group at the 4-position of the benzene ring. These derivatives 5a-5i and 5k were evaluated for GLS1 inhibitory activity (Fig. 3(B)).

As shown in FIG. 3(B), all derivatives were found to have GLS1 inhibitory activity. Among them, the derivative 5i in which the hydrogen atom at the 4-position of the benzene ring was substituted with a fluorine atom had the strongest GLS1 inhibitory activity, and its Glu production rate at a concentration of 10 mM was 90% lower than that of the control. However, the GLS1 inhibitory activity of the derivative 5i was lower than that of the amide derivative 2j, suggesting that introduction of a substituent to the benzene ring tended to lower the GLS1 inhibitory activity.

(3) Increase/Decrease in Carbon Number of Linker Between Benzene Ring and Pyrazole Ring Two derivatives 8a and 8b were synthesized in order to examine the effect of increasing/decreasing the carbon number in the linker between the benzene ring and the pyrazole ring. Specifically, as shown in Scheme 3, first, phenylmethylketone 6a and benzylacetone 6b were used, and in the same manner as in Scheme 2 above, an amide derivative 8a in which a benzene ring and a pyrazole ring, respectively, were directly bonded, and both Converted to amide 8b with two linker carbons (ethylene groups) between the rings.

These amide derivatives (8a and 8b) were evaluated for GLS1 inhibitory activity (FIGS. 3(A) and (B)). As shown in FIG. 3(B), both amide derivatives 8a and 8b had GLS1 inhibitory activity. However, the GLS1 inhibitory activity of the amide derivative 8b was lower than that of the amide derivative 8a, suggesting that the GLS1 inhibitory activity depends on the carbon number (length) of the linker. It was observed that the smaller the carbon number (n number) of the linker, preferably n=0, the higher the GLS1 inhibitory activity.

(4) Substitution of Pyrazole Ring to Other Heterocycle As shown in Scheme 4 below, imidazole derivative 10 was synthesized from known ester compound 8 prepared from 2-bromoacetophenone.

Further, as shown in Scheme 5 below, pyrrole derivatives 13 and 14 were similarly synthesized from known ester compounds 10 and 12, respectively.

These derivatives 13 and 14 were evaluated for GLS1 inhibitory activity (Fig. 3(B)).
As shown in FIG. 3(B), the imidazole derivative 10 had relatively high GLS1 inhibitory activity, although slightly inferior to the corresponding pyrazole derivative 8a. On the other hand, pyrrole derivatives 13 and 14 corresponding to these were both found to significantly reduce the GLS1 inhibitory activity. This suggests that a heterocyclic ring having two nitrogen atoms (pyrazole ring or imidazole ring) is important for the GLS1 inhibitory activity as the heterocyclic ring forming the basic skeleton.

From the above results, as shown in FIG. 3(B), particularly high GLS1 inhibitory activity was observed in pyrazole derivatives 2j, 5i and 8a among the compounds evaluated above.

Experimental Example 3: Evaluation of GLS1 inhibitory activity using recombinant GLS1 Pyrazole derivatives 5i and 8a, which were confirmed to have high GLS1 inhibitory activity in Experimental Example 2, were re-evaluated for their GLS1 inhibitory activity using mouse and human recombinant GLS1. (Figures 4 and 5).
In the evaluation using the mouse recombinant GLS1, the positive controls DON and CB-839 had an IC ₅₀ of approximately 1 mM for DON and between 10 and 100 nM for CB-839, consistent with previous reports. As shown in FIG. 4, both pyrazole derivatives 5i and 8a significantly inhibited Glu production at 10 mM, which had inhibitory activity in mouse kidney extract, and IC ₅₀ was confirmed to be between 1 and 10 mM. Ta. Moreover, as shown in FIG. 5, it was confirmed that all of these compounds significantly suppressed Glu production even in human recombinant GLS1 at 10 mM, exhibiting high GLS1 inhibitory activity.

Experimental Example 3: Evaluation of anticancer activity
It has been pointed out that GLS1 is associated with the progression of various cancers, and it is known that inhibiting glutamine metabolism by inhibiting GLS1 suppresses the growth and proliferation of cancer cells. Various compounds with GLS1 inhibitory activity are currently undergoing clinical trials in cancer patients with renal cell carcinoma, colon cancer, non-small cell lung cancer, melanoma, and progressive myelodysplastic syndrome. It is expected to have a therapeutic effect on various cancer types.

Therefore, as representative compounds having GLS1 inhibitory activity confirmed in Experimental Example 2, the pyrazole derivatives 5i and 8a having particularly high GLS1 inhibitory activity were used to evaluate the growth inhibitory effect on cancer cells. As cancer cells, a human mammary adenocarcinoma-derived cell line (MCF7) and an acute myeloid leukemia-derived cell line (MOLM13) were used.

In the experiment, pyrazole derivative 5i or 8a was added to each cell (3 × 10 ³ cells/ml) at various concentrations and cultured. The cell growth inhibitory effect was evaluated from the ratio of DMSO added instead of the derivative) to the number of cells (100%) (n=3). For MCF7, the medium was changed on the first day, the third day, and the sixth day of culture.

Results for a human breast adenocarcinoma-derived cell line (MCF7) and an acute myeloid leukemia-derived cell line (MOLM13) are shown in Figures 6 and 7, respectively.
As shown in FIG. 6, pyrazole derivatives 5i and 8a inhibited cell proliferation by 40% and 67%, respectively, at 100 μM against MCF7 cells (IC ₅₀ [μM]: 5i=77.5, 8a=93.1). Moreover, as shown in FIG. 7, pyrazole derivatives 5i and 8a at 100 μM inhibited cell growth of MOLM13 cells by 60% and 92.5%, respectively (IC ₅₀ [μM]: 5i=58.6, 8a=62.1).

As shown in these results, the compounds having GLS1 inhibitory activity confirmed in Experimental Example 2 act to suppress the proliferation of cancer cells based on their GLS1 inhibitory action, as typified by pyrazole derivatives 5i and 8a. It was confirmed that it is effective for anticancer therapy such as prevention and treatment of cancer, suppression of cancer progression, and prevention of cancer recurrence.

SEQ ID NO: 1 is the base sequence of primer G1U1 (NdeI site) used for the production of mouse recombinant GLS1, and SEQ ID NO: 2 is the base sequence of primer G1-204L1 (BamHI site) used for the same production.

Claims (12)

A glutaminase 1 inhibitor comprising a compound represented by general formula (I), a pharmaceutically acceptable salt thereof, or a solvate thereof as an active ingredient:

[In formula (I),
X is _CH2 or C=O;
n means an integer of 0-2. However, n is 1 when X is C=O.
Y is a divalent 5-membered heterocyclic group having at least two nitrogen atoms;
R ¹ is a hydrogen atom, a halogen atom, a haloalkyl group, an alkoxy group, an alkyl group, or a hydroxyl group;
R ² is a group represented by any one of the following formulas (a) to (c):

(In formula (a),
A means a hydroxyl group or an oxygen atom,
A double line consisting of a dotted line and a solid line means a single bond when A is a hydroxyl group and a double bond when it is an oxygen atom,
The asterisk denotes the point of attachment to the carbon atom of the carbonyl group in formula (I). );

(In formula (b),
B and D are the same or different and represent a hydrogen atom, an alkyl group, or an alkoxy group;
The asterisk denotes the point of attachment to the carbon atom of the carbonyl group in formula (I). );

(In formula (c),
E is an oxygen atom or an alkylene group in which one of the hydrogen atoms may be substituted with a hydroxyl group,
The asterisk denotes the point of attachment to the carbon atom of the carbonyl group in formula (I). );
When n is 0, R ₁ is not an alkoxy group, and when R ₁ is a hydroxyl group, R ² is a group represented by formula (b) or (c). ]. 2. The glutaminase-1 inhibitor according to claim 1, wherein in the compound represented by formula (I), Y is a pyrazole group or an imidazole group. In the compound represented by the formula (I), R ² is a group represented by the formula (a),
In formula (a), A is a hydroxyl group, a double line consisting of a dotted line and a solid line is a single bond,
X is CH ₂ and n is an integer from 0 to 2;
The glutaminase-1 inhibitor according to claim 1 or 2. In the compound represented by the formula (I), R ² is a group represented by the formula (b),
X is CH ₂ and n is an integer from 0 to 2;
The glutaminase-1 inhibitor according to claim 1 or 2. In the compound represented by the formula (I), R ² is a group represented by the formula (c),
X is CH ₂ and n is 0-2;
The glutaminase-1 inhibitor according to claim 1 or 2. A pharmaceutical composition, food composition or cosmetic composition comprising the glutaminase 1 inhibitor according to any one of claims 1 to 5 as an active ingredient. 7. The pharmaceutical composition, food composition or cosmetic composition according to claim 6, which is used for prevention, amelioration or treatment of inflammation, diseases derived from inflammation, obesity, metabolic syndrome, aging or cancer. A compound having a glutaminase 1 inhibitory action represented by the general formula (I), a pharmaceutically acceptable salt thereof, or a solvate thereof:

[In formula (I),
X is _CH2 or C=O;
n means an integer of 0-2. However, n is 1 when X is C=O.
Y is a divalent 5-membered heterocyclic group having at least two nitrogen atoms;
R ¹ is a hydrogen atom, a halogen atom, a haloalkyl group, an alkoxy group, an alkyl group, or a hydroxyl group;
R ² is a group represented by any one of the following formulas (a) to (c):

(In formula (a),
A means a hydroxyl group or an oxygen atom,
A double line consisting of a dotted line and a solid line means a single bond when A is a hydroxyl group and a double bond when it is an oxygen atom,
The asterisk denotes the point of attachment to the carbon atom of the carbonyl group in formula (I). );

(In formula (b),
B and D are the same or different and represent a hydrogen atom, an alkyl group, or an alkoxy group;
The asterisk denotes the point of attachment to the carbon atom of the carbonyl group in formula (I). );

(In formula (c),
E is an oxygen atom or an alkylene group in which one of the hydrogen atoms may be substituted with a hydroxyl group,
The asterisk denotes the point of attachment to the carbon atom of the carbonyl group in formula (I). );
When n is 0, R ₁ is not an alkoxy group, and when R ₁ is a hydroxyl group, R ² is a group represented by formula (b) or (c).
However, N-(4-hydroxycyclohexyl)-3-phenyl-1H-pyrazole-5-carboxamide is excluded. ]. 9. A compound according to claim 8, wherein Y is a pyrazole group or an imidazole group. wherein R ² is a group represented by formula (a),
In formula (a), A is a hydroxyl group, a double line consisting of a dotted line and a solid line is a single bond,
X is CH ₂ and n is an integer from 0 to 2;
10. A compound according to claim 8 or 9. In the compound represented by the formula (I), R ² is a group represented by the formula (b),
X is CH ₂ and n is an integer from 0 to 2;
10. A compound according to claim 8 or 9. In the compound represented by the formula (I), R ² is a group represented by the formula (c),
X is CH ₂ and n is 0-2;
10. A compound according to claim 8 or 9.

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