Glutaminase 1 inhibitor

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]

[0001] The present invention relates to a glutaminase 1 inhibitor and a pharmaceutical composition, a food or drink composition, or a cosmetic composition containing the same as an active ingredient. The present invention also relates to a novel compound useful as an active ingredient of a glutaminase 1 inhibitor. [Background technology]

[0002] Glutaminase is an enzyme that produces glutamic acid from glutamine. There are two types of glutaminase: 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 will be abbreviated as "GLS1," glutaminase 2 as "GLS2," and both will be collectively referred to as "GLS."

[0003] GLS1 is widely present in the kidney, small intestine, and brain, while 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 controlling many processes, such as metabolism, protein synthesis and decomposition, and insulin resistance (Non-Patent Document 1). GLS has also been found to be overexpressed in various cancer cells (e.g., triple-negative breast cancer (TNBC) cells and hepatocellular carcinoma (HCC) cells), and GLS inhibitors such as CB-839 that target GLS have been developed as medicines (Non-Patent Document 2). GLS inhibitors such as CB-839 have an anti-inflammatory effect and are known to be useful as anti-inflammatory agents and for preventing, improving, or treating diseases caused by inflammation (Patent Document 1). Furthermore, the GLS inhibitors have an anti-obesity effect and are known to be useful as anti-obesity agents for preventing, improving, or treating metabolic syndrome (Patent Document 2). Furthermore, recent studies have suggested that inhibition of GLS1 has the effect of preventing aging (Non-Patent Document 3). Senescent cells increase GLS1 and regulate intracellular pH homeostasis by producing excess ammonia when intracellular pH drops. Therefore, it is expected that GLS1 inhibitors will induce apoptosis in senescent cells, improving various symptoms such as glomerular sclerosis in the kidney, pulmonary fibrosis, and inflammatory cell infiltration in the liver. Furthermore, it has been shown that GLS1 inhibitors are also effective in increasing muscle mass and suppressing fat atrophy associated with aging (Non-Patent Document 3). Thus, GLS1 inhibitors are expected to be used as anti-aging drugs.

[0004] Thus, the importance of GLS as a drug target is increasing today, and new GLS inhibitors are needed.

[0005] In addition to the above-mentioned CB-839, 6-diazo-5-oxo-L-norleucine (hereinafter, also simply referred to as "DON") and bis-2-(5-phenylacetamido-1,2,4-thiadiazol-2-yl)ethyl sulfide (hereinafter, also simply referred to as "BPTES") are known as GLS inhibitors. However, DON has a problem in that it not only acts on cancer cells by acting on the substrate binding site of glutaminase, but also acts on other enzymes that use glutamine as a substrate (Non-Patent Documents 4 and 5). BPTES exhibits stronger GLS1 inhibitory activity than DON by specifically binding to the allosteric site of glutaminase, but is unsuitable as a pharmaceutical product due to its low solubility (Non-Patent Document 6). [Prior art documents] [Patent documents]

[0006] [Patent Document 1] JP 2020-29412 A [Patent Document 2] JP 2020-28239 A [Non-patent literature]

[0007] [Non-Patent Document 1] Molecular mechanisms of glutamine action, J. Cell. Physiol., 2005, 204(2): 392-401. [Non-Patent Document 2] Antitumor activity of the Glutaminase inhibitor CB-839 in triple-negative breast cancer. Mol Cancer Ther. 2014 Apr; 13 (4):890-901. [Non-Patent Document 3] Senolysis by Glutaminolysis inhibition ameliorates various age-associated disorders. Science. 2021 Jan 15; 371 (6526):265-270.Senolysis by glutaminolysis inhibition ameliorates various age-associated disorders. Science 2021, 371, 265-270. [Non-Patent Document 4] Metabolism and action of amino acid analog anti-cancer agents. Pharmacology & therapeutics 1990, 46, 243-271. [Non-Patent Document 5] Azaserine, DON, and azotomycin: three diazo analogs of L-glutamine with clinical antitumor activity. Cancer treatment reports 1979, 63, 1033-1038 [Non-Patent Document 6] Novel mechanism of inhibition of rat kidney-type glutaminase by bis-2-(5-phenylacetamido-1,2,4-thiadiazol-2-yl)ethyl sulfide (BPTES). The Biochemical journal 2007, 406, 407-414. Summary of the Invention [Problem to be solved by the invention]

[0008] An objective of the present invention is to provide a new GLS1 inhibitor. Another objective of the present invention is to provide a pharmaceutical composition, food or drink composition, or cosmetic composition that is useful for preventing or ameliorating diseases or pathological conditions that are effectively treated by inhibiting GLS1. A further object of the present invention is to provide novel compounds that are useful as GLS1 inhibitors or as active ingredients of the above-mentioned pharmaceutical compositions, food and beverage compositions, or cosmetic compositions. [Means for solving the problem]

[0009] In order to solve the above problems, the present inventors searched for candidate GLS1 inhibitor compounds from a commercially available compound database by multistage in silico screening (a primary screening using a 3D pharmacophore obtained from the complex crystal structure of glutaminase and a known GLS1 inhibitor, filtering by Lipinski's "Rule of Five", a secondary screening using molecular docking calculations, filtering by binding free energy calculations, filtering based on docking poses, and in silico screening consisting of molecular similarity analysis based on chemical structures), and found that the thiadiazole compound represented by the following formula has a stronger GLS1 inhibitory effect than DON. [ka]

[0010] Based on the above findings, the present invention was completed as a result of further investigations to find a compound that has GLS1 inhibitory activity similar to the above compound, and has the following embodiments.

[0011] (I) GLS1 inhibitor (I-1) A GLS1 inhibitor comprising a compound represented by general formula (I), a pharma- ceutically acceptable salt thereof, or a solvate thereof as an active ingredient: [ka] [In formula (I), X is an optionally substituted 5-membered heterocyclic group having at least two nitrogen atoms; Y is -NH-, -C(=O)-, -(CH2) mA divalent group formed by linking two or three groups selected from the group consisting of -, -, and -O-; R 1 and R 2 are the same or different and are an alkoxy group or a hydrogen atom; or R 1 and R 2 are linked together to form a dioxyalkylene group and are bonded to adjacent carbon atoms on the phenyl ring to form a 5- or 6-membered fused ring; R 3 represents an optionally substituted phenyl group, an optionally substituted cycloalkyl group, or an alkyl group; n is 0 or 1, and m is 1, 2 or 3.

[0012] (I-2) The GLS1 inhibitor according to (I-1), wherein, in the compound represented by formula (I), X is a thiadiazole residue or a triazole residue, preferably a thiadiazole residue.

[0013] (I-3) A GLS1 inhibitor according to (I-1) or (I-2), wherein in the compound represented by formula (I), Y is a group selected from the group consisting of -NH-C(=O)-, -NH-C(=O)-CH2-, -C(=O)-NH-CH2-, and -CH2-O-, preferably -NH-C(=O)-. (I-4) In the compound represented by the formula (I), R 1 and R 2 A GLS1 inhibitor according to any one of (I-1) to (I-3), wherein one of said groups is an alkoxy group and the other is a hydrogen atom. (I-5) In the compound represented by the formula (I), R 3 is a phenyl group which may have a substituent, preferably a phenyl group which has a substituent. (I-6) The GLS1 inhibitor according to any one of (I-1) to (I-5), wherein in the compound represented by formula (I), n is 1.

[0014] (II) Pharmaceutical compositions, food and beverage compositions, or cosmetic compositions (II-1) A pharmaceutical composition, a food or drink composition, or a cosmetic composition, which contains as an active ingredient a GLS1 inhibitor according to any one of (I-1) to (I-6). (II-2) A pharmaceutical composition, food or beverage composition, or cosmetic composition according to (II-1), which is used for the prevention, amelioration, or treatment of inflammation, diseases resulting from inflammation, obesity, metabolic syndrome, aging, or cancer.

[0015] (III) Compounds having GLS1 inhibitory activity (III-1) A compound having a GLS1 inhibitory effect represented by general formula (II), a pharma- ceutically acceptable salt thereof, or a solvate thereof: [ka] [In formula (II), R 1 and R 2 are the same or different and each is an alkoxy group or a hydrogen atom; R 3 represents a phenyl group, a cycloalkyl group, or an alkyl group, each of which may have a substituent.

[0016] (III-2) R 1 and R 2 is a hydrogen atom, or a pharma- ceutically acceptable salt or solvate thereof, according to (III-1).

[0017] (III-3) The compound having GLS1 inhibitory activity is the compound described below in (III-1), its pharma- ceutically acceptable salt, or a solvate thereof; (a) Compound 1: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)cyclobutanecarboxamide. (b) Compound 3: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)cyclohexanecarboxamide. (c) Compound 5: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)isobutyramide. (d) Compound 7: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-fluorobenzamide. (e) Compound 8: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-chlorobenzamide. (f) Compound 9: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-methylbenzamide. (g) Compound 11: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-nitrobenzamide. (h) Compound 12: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-methoxybenzamide. (i) Compound 15: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-2-(trifluoromethyl)benzamide. (j) Compound 16: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-3-(trifluoromethyl)benzamide. (k) Compound 17: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-2-fluorobenzamide. (l) Compound 18: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-3-fluorobenzamide. (m) Compound 20: N-(5-((2-((4-Methoxybenzyl)amino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)isobutyramide Effect of the Invention

[0018] According to the present invention, a novel GLS1 inhibitor can be provided. The GLS1 inhibitor of the present invention is useful as an active ingredient of a pharmaceutical composition, a food or beverage composition, or a cosmetic composition for preventing or ameliorating diseases or pathological conditions that are effectively treated by inhibiting GLS1 (e.g., inflammation, diseases or pathological conditions caused by inflammation, obesity, metabolic syndrome, aging, cancer, etc.). Therefore, the pharmaceutical composition, food and beverage composition, or cosmetic composition of the present invention, which contains a GLS1 inhibitor as an active ingredient, can be effectively used to prevent or ameliorate, for example, inflammation, diseases and pathological conditions resulting from inflammation, obesity, metabolic syndrome, aging, or cancer, by inhibiting GLS1. Furthermore, the compounds provided by the present invention are expected to inhibit GLS1 and thereby exhibit effects of preventing or ameliorating, for example, inflammation, diseases and pathologies resulting from inflammation, obesity, metabolic syndrome, aging, or cancer, and are useful as GLS1 inhibitors, and as active ingredients of pharmaceutical compositions, food and beverage compositions, or cosmetic compositions. [Brief description of the drawings]

[0019] [Figure 1-1] 1 shows the structural formulas of candidate GLS1 inhibitor compounds A to E whose GLS1 inhibitory activity was evaluated in Experimental Example 1. [Figure 1-2] FIG. 1 shows the results of evaluating the GLS1 inhibitory activity of a known GLS1 inhibitor (DON) and compounds A to E, as performed in Experimental Example 1. The vertical axis shows the measurement results (glutamic acid concentration) of each compound as a relative percentage (%) to the measurement result (100%) of the control (solvent DMSO). [Diagram 2]FIG. 1 shows the results of evaluating the GLS1 inhibitory activity of DON, compound E, and compounds 1 to 24, performed in Experimental Example 2. The vertical axis shows the measurement results (glutamic acid concentration) of each compound as a relative percentage (%) to the measurement result (100%) of the control (solvent DMSO). [Diagram 3] In Experimental Example 2, the experimental results of determining the IC50 of the GLS1 inhibitory activity for DON, compound E, and compound 23 are shown. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0020] (I) GLS1 inhibitor The GLS1 inhibitor of the present invention is characterized in that it contains as an active ingredient a compound represented by the following formula (I) (hereinafter also referred to as "compound (I)"), a pharma- ceutically acceptable salt thereof, or a solvate thereof: [ka]

[0021] In formula (I), X is an optionally substituted 5-membered heterocyclic group having at least two nitrogen atoms, which is hereinafter also referred to simply as an "N-containing 5-membered heterocyclic group." The N-containing 5-membered heterocyclic group includes a 5-membered heterocyclic group having three nitrogen atoms in the ring and the remainder being carbon atoms; and a 5-membered heterocyclic group having two nitrogen atoms and one sulfur atom in the ring and the remainder being carbon atoms.

[0022] Preferably, it is an N-containing 5-membered unsaturated heterocyclic group. Such groups include a triazole residue, which is a 5-membered unsaturated heterocyclic group having three nitrogen atoms and two carbon atoms in the ring, and a thiadiazole residue, which is a 5-membered unsaturated heterocyclic group having two nitrogen atoms, one sulfur atom, and two carbon atoms in the ring. Preferably, it is a thiadiazole residue.

[0023] The triazole residue includes a 1,2,4-triazole residue in which the 1st, 2nd and 4th positions of the ring are nitrogen atoms, and a 1,2,3-triazole residue in which the 1st, 2nd and 3rd positions of the ring are nitrogen atoms. In the 1,2,4-triazole residue, the atoms (two bonds) bonding to adjacent groups are two carbon atoms, and in the 1,2,3-triazole residue, the bonds are a nitrogen atom and a carbon atom, and these are preferably located as far apart as possible.

[0024] The thiadiazole residue includes a 1,3,4-thiadiazole residue in which the 1st position of the ring is a sulfur atom, the 3rd and 4th positions are nitrogen atoms, and the other two carbon atoms are atoms (bonds) that bond to adjacent groups; a 1,2,4-thiadiazole residue in which the 1st position of the ring is a sulfur atom, the 2nd and 4th positions are nitrogen atoms, and the other two carbon atoms are bonds to adjacent groups; a 1,2,3-thiadiazole residue in which the 1st position of the ring is a sulfur atom, the 2nd and 3rd positions are nitrogen atoms, and one of the nitrogen atoms and one of the other carbon atoms are bonds to adjacent groups; and a 1,2,5-thiadiazole residue in which the 1st position of the ring is a sulfur atom, the 2nd and 5th positions are nitrogen atoms, and one of the nitrogen atoms and one of the other carbon atoms are bonds to adjacent groups. As the thiadiazole residue, a 1,3,4-thiadiazole residue in which the atom that bonds to the adjacent group is a carbon atom, and a 1,2,4-thiadiazole residue are preferred, and a 1,3,4-thiadiazole residue is more preferred.

[0025] These 5-membered heterocyclic groups may have a substituent. There is no particular limitation as to whether or not they have a substituent, but when the 5-membered heterocyclic group is a triazole residue, it is preferable that it has a substituent, and when the 5-membered heterocyclic group is a thiadiazole residue, it is preferable that it is unsubstituted. Substituents include, but are not limited to, alkyl groups and amino groups, as long as compound (I) has GLS1 inhibitory activity. Examples of the alkyl groups include linear or branched alkyl groups having 1 to 6 carbon atoms (C1 to 6). More preferred are C1 to 3 alkyl groups. These alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, and n-hexyl groups. Although not limited to, preferred are methyl, ethyl, and tert-butyl groups, and more preferred is methyl.

[0026] Y is -NH-, -C(=O)-, -(CH2) m A divalent group formed by combining two or three selected from the group consisting of -, -, and -O-. Examples of combinations thereof include, but are not limited to, -NH-C(=O)-, -NH-C(=O)-(CH2) m -, -C(=O)-NH-, -(CH2) m -C(=O)-NH- and -(CH2) m An example is -O-. When X is a thiadiazole residue, Y is preferably -NH-C(=O)- and -NH-C(=O)-(CH2) m -; more preferably -NH-C(=O)-. When X is a triazole residue, Y is preferably -(CH2) m -C(=O)-NH- and -(CH2) m -O-. Here, m is an integer of 1 to 3, preferably 1 or 2, and more preferably 1.

[0027] R 1 and R 2 are the same or different and are an alkoxy group or a hydrogen atom; or R 1 and R 2 are linked together to form a dioxyalkylene group, which are bonded to adjacent carbon atoms on the phenyl ring to form a 5- or 6-membered fused ring.

[0028] R 1 and R 2 The alkoxy group represented by the formula (I) is a group having a structure in which an alkyl group is bonded to an oxygen atom. The types of alkyl groups contained in this structure can be the same as those of the substituents of the 5-membered heterocyclic group explained above. It is preferably a C1-3 alkyl group. The preferred alkoxy groups are a methoxy group and an ethoxy group, and more preferably a methoxy group.

[0029] R 1 and R 2 When these are linked to each other to form a dioxyalkylene group, examples of the dioxyalkylene group include a dioxymethylene group and a dioxyethylene group. A dioxymethylene group is preferred.

[0030] In formula (I), R on the phenyl ring 1 and R 2 The positions of R may be any of ortho, meta, or para positions, as long as compound (I) has GLS1 inhibitory activity, but preferably, one is meta and the other is para. Although not limited thereto, when the position of the bond to the methylene group of the phenyl ring in compound (I) is the 1st position, R may be at the 3rd position (counterclockwise). 1 Ki, R in 4th place 2 It is preferred that the group is coordinated.

[0031] R 3 is a phenyl group or a cycloalkyl group, which may have one or more substituents, or an alkyl group.

[0032] The cycloalkyl group includes a cycloalkyl group having 3 to 6 carbon atoms. Specific examples include a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, and a cyclohexyl group. A cycloalkyl group having 4 to 6 carbon atoms is preferable.

[0033] Examples of the alkyl group include linear or branched alkyl groups having 1 to 6 carbon atoms (C1 to 6). A linear or branched alkyl group having 3 to 6 carbon atoms is preferred. Although not limited thereto, preferred are an isopropyl group, an n-butyl group, a sec-butyl group, an isobutyl group, a tert-butyl group, and a pentyl group, and more preferred is an isopropyl group.

[0034] R 3 When is a phenyl group or a cycloalkyl group, these functional groups may have one or more, preferably one or two, substituents, as long as compound (I) has GLS1 inhibitory activity. The substituents include a haloalkyl group, a haloalkoxy group, a halogen atom, an alkoxy group, an alkyl group, a nitro group, and a hydroxyl group. When X is a thiadiazole residue, the substituent on the phenyl group is preferably a haloalkyl group, a haloalkoxy group, or a halogen atom.When X is a triazole residue, the substituent on the phenyl group is preferably an alkyl group.

[0035] The alkyl group is preferably, but not limited to, a linear or branched alkyl group having 1 to 6 carbon atoms (C1 to 6). More preferably, it is a C1 to 3 alkyl group. These alkyl groups include a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a tert-butyl group, an n-pentyl group, and an n-hexyl group. Although not limited to, it is preferably a methyl group, an ethyl group, and a tert-butyl group, and more preferably, it is a methyl group.

[0036] An alkoxy group 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, and is preferably a C1-3 alkyl group. Specific examples of the alkoxy group include a methoxy group and an ethoxy group. A methoxy group is preferred.

[0037] The halogen atom includes, for example, a fluorine atom, a chlorine atom, a bromine atom and an iodine atom, and is preferably a fluorine atom or a chlorine atom.

[0038] The haloalkyl group includes a straight-chain or branched alkyl group having 1 to 6 carbon atoms (C1 to 6) and 1 to 3 halogen atoms. A trihaloC1 to 6 alkyl group having 3 halogen atoms is preferred, and a trihaloC1 to 3 alkyl group is more preferred. Examples of the trihaloC1 to 6 alkyl group include a trifluoromethyl group, a trichloromethyl group, a 2,2,2-trifluoroethyl group, a 2,2,2-trichloroethyl group, and a 3,3,3-trifluoropropyl group. A trifluoromethyl group is preferred.

[0039] The haloalkoxy group includes a straight-chain or branched alkoxy group having 1 to 3 halogen atoms and 1 to 6 carbon atoms (C1 to 6). A trihaloC1 to 6 alkoxy group having 3 halogen atoms is preferred, and a trihaloC1 to 3 alkoxy group is more preferred. Examples of the trihaloC1 to 6 alkoxy group include a trifluoromethytoxy group, a trichloromethoxy group, a 2,2,2-trifluoroethoxy group, a 2,2,2-trichloroethoxy group, and a 3,3,3-trifluoropropoxy group. A trifluoromethoxy group is preferred.

[0040] R 3 When is a phenyl group, the coordination position of the substituent may be any of the ortho-position, meta-position, and / or para-position, as long as compound (I) has GLS1 inhibitory activity. Although not limited, the coordination position is preferably the para-position. R 3 When R is a cycloalkyl group, the coordination position of the substituent can be selected arbitrarily, as long as compound (I) has GLS1 inhibitory activity. 3 It is preferable that the carbon atom in the alkyl group is coordinated to a carbon atom (non-adjacent carbon atom) that is at least one atom, preferably two or three atoms, away from the bond (carbon atom) to the carbonyl group in the alkyl group.

[0041] In formula (I), n is 0 or 1. Preferably, it is 1. In particular, when X is a thiadiazole residue, n is preferably 1.

[0042] Compound (I) includes compounds represented by general formulas (III) to (V) shown in (A) to (C) below:

[0043] (A) Compounds in which X is a thiadiazole residue in the general formula (I) (collectively referred to as "thiadiazole compounds"): [ka] [In formula (III), Y, R 1 , R 2 , R 3、 and n are as defined above.]

[0044] The thiadiazole compounds preferably include the following compounds: (A1) In formula (III), Y is -NH-C(=O)-, R 1 and R 2 is a hydrogen atom, R 3 A compound in which is a substituted phenyl group and n is 1. (A2) In formula (III), Y is -NH-C(=O)-, R 1 and R 2 is a hydrogen atom, R 3 is a substituted or unsubstituted cycloalkyl group, and n is 1. (A3) In formula (III), Y is -NH-C(=O)-, R 1 and R 2 is a hydrogen atom, R 3 A compound in which is an alkyl group and n is 1. (A4) In formula (III), Y is -NH-C(=O)-, R 3 is a phenyl group having a substituent, R 1 and R 2 At least one of the above is an alkoxy group and n is 1. (A5) In formula (III), Y is -NH-C(=O)-, R 3 is an alkyl group, R 1 and R 2 At least one of the above is an alkoxy group and n is 1. (A6) In formula (III), Y is -NH-C(=O)-(CH2) m -, R 1 and R 2 are linked together to form a dioxyalkylene group, and are linked to adjacent carbon atoms on the phenyl ring to form a 5- or 6-membered fused ring; R 3 is an optionally substituted cycloalkyl group or phenyl group, n is 1, and m is 0 or 1. Among these, thiadiazole compounds included in groups (A1), (A3), (A4) and (A6) are preferred.

[0045] The groups (A1) to (A6) specifically include the following compounds. Regardless of stereoisomers, the compounds also include isomers. Thiadiazole compounds included in group (A1) : Compound 7: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-fluorobenzamide. Compound 8: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-chlorobenzamide. Compound 9: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-methylbenzamide. Compound 10: N-(5-((2-(benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-(trifluoromethyl)benzamide. Compound 11: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-nitrobenzamide. Compound 12: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-methoxybenzamide. Compound 13: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-hydroxybenzamide. Compound 14: N-(5-((2-(benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-(trifluoromethoxy)benzamide. Compound 15: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-2-(trifluoromethyl)benzamide. Compound 16: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-3-(trifluoromethyl)benzamide. Compound 17: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-2-fluorobenzamide. Compound 18: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-3-fluorobenzamide.

[0046] Thiadiazole compounds included in group (A2) : Compound 1: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)cyclobutanecarboxamide. Compound 2: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)cyclopentanecarboxamide. Compound 3: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)cyclohexanecarboxamide. Compound 4: trans-N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-hydroxycyclohexanecarboxamide.

[0047] Thiadiazole compounds included in group (A3) : Compound 5: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)isobutyramide. Compound 6: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)hexanamide.

[0048] Thiadiazole compounds included in group (A4) : Compound 22: N-(5-((2-((3-Methoxybenzyl)amino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-(trifluoromethyl)benzamide. Compound 23: N-(5-((2-((4-Methoxybenzyl)amino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-(trifluoromethyl)benzamide. Compound 24: N-(5-((2-((3,4-Dimethoxybenzyl)amino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-(trifluoromethyl)benzamide

[0049] Thiadiazole compounds included in group (A5) : Compound 19: N-(5-((2-((3-Methoxybenzyl)amino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)isobutyramide Compound 20: N-(5-((2-((4-Methoxybenzyl)amino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)isobutyramide Compound 21: N-(5-((2-((3,4-Dimethoxybenzyl)amino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)isobutyramide

[0050] Thiadiazole compounds included in group (A6) : Compound E: N-(5-((2-((benzo[d][1,3]dioxol-5-ylmethyl)amino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)cyclobutanecarboxamide Compound C: N-(benzo[d][1,3]dioxol-5-ylmethyl)-2-((5-(2-phenylacetamido)-1,3,4-thiadiazol-2-yl)thio)acetamide

[0051] (B) Compounds in which X is a 1,2,3-triazole residue in the general formula (I) (collectively referred to as "triazole compound 1"): [ka] [In formula (IV), Y, R 1 , R 2 , R 3 , and n are as described above.4 is a hydrogen atom, an amino group, or an alkyl group having 1 to 3 carbon atoms.]

[0052] The triazole compound 1 preferably includes the following compounds: Regardless of stereoisomers, the compounds also include all isomers. (B1) In formula (IV), Y is -(CH2)-C(=O)-NH-, R 3 is a phenyl group having one or two substituents, R 1 and R 2 are linked together to form a dioxyalkylene group, and are linked to adjacent carbon atoms on the phenyl ring to form a 5- or 6-membered fused ring; R 4 A compound in which is an amino group and n is 0. Compound A: 5-amino-N-(benzo[d][1,3]dioxol-5-ylmethyl)-1-(2-oxo-2-(o-tolylamino)ethyl)-1H-1,2,3-triazole-4-carboxamide Compound B: 5-amino-N-(benzo[d][1,3]dioxol-5-ylmethyl)-1-(2-((3,4-dimethylphenyl)amino)-2-oxoethyl)-1H-1,2,3-triazole-4-carboxamide

[0053] (C) Compounds in which X is a 1,2,4-triazole residue in the general formula (I) (collectively referred to as "triazole compounds 2"): [ka] [In formula (V), Y, R 1 , R 2 , R 3 , and n are as described above. 5is a hydrogen atom or an alkyl group having 1 to 3 carbon atoms.]

[0054] The triazole compound 2 preferably includes the following compounds: Regardless of stereoisomers, the compounds also include all isomers. (C1) In formula (V), Y is -CH2-O-, R 3 is a phenyl group having one or two substituents, R 1 and R 2 are linked together to form a dioxyalkylene group, and are linked to adjacent carbon atoms on the phenyl ring to form a 5- or 6-membered fused ring; R 5 is an alkyl group having 1 to 3 carbon atoms (preferably a methyl group); and n is 1. Compound D: N-(benzo[d][1,3]dioxol-5-ylmethyl)-2-((5-((2,3-dimethylphenoxy)methyl)-4-methyl-4H-1,2,4-triazol-3-yl)thio)acetamide

[0055] The aforementioned compound (I) can take the form of a pharma- ceutically acceptable salt, and the salt can be used in the same manner as the free form of compound (I). The pharma- ceutically acceptable salt includes salts with inorganic acids such as hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, or boric acid (inorganic acid salts); salts with organic acids such as formic acid, acetic acid, lactic acid, fumaric acid, maleic acid, tartaric acid, citric acid, succinic acid, malonic acid, or tosylic acid (organic acid salts). However, the salts are not limited thereto. These salts can be prepared by conventional means. The aforementioned compound (I) and its pharma-ceutically acceptable salts may be solvates. The solvates also include hydrates.

[0056] Compound (I), a pharma- ceutically acceptable salt thereof, or a solvate thereof (hereinafter, these are also collectively referred to as "compound (I) type") can be produced according to the description of the production examples described below.

[0057] The compound (I) obtained by these production methods is isolated and purified from the reaction mixture by applying known isolation and / or purification means, such as distillation, recrystallization, solvent extraction, column chromatography, ion exchange chromatography, gel chromatography, affinity chromatography, preparative thin layer chromatography, etc.

[0058] Compound (I) has GLS1 inhibitory activity, as shown in Experimental Examples 1 to 3 described below.

[0059] The GLS1 inhibitor according to the present invention may consist of only the above-mentioned compound (I), but may also contain other components such as carriers and additives that are commonly used in the relevant fields (biochemistry, medicine, cosmetics, and food). The specific types of carriers and additives and their blending ratios are not particularly limited, as long as the GLS1 inhibitor exerts GLS1 inhibitory activity based on the compound (I). For example, the types of carriers and additives and their blending ratios can be appropriately selected with reference to the types and blending ratios of the carriers and additives described in the pharmaceutical composition described below.

[0060] (II) Pharmaceutical Composition The pharmaceutical composition of the present invention (hereinafter referred to as "the pharmaceutical composition") is characterized by containing the aforementioned GLS1 inhibitor, specifically, the aforementioned compound (I), a pharma- ceutical acceptable salt thereof, or a solvate thereof, as an active ingredient.

[0061] Based on the GLS1 inhibitory activity of the above-mentioned compound (I), the pharmaceutical composition can be applied to diseases or symptoms (including conditions) for which inhibition of GLS1 is effective. Diseases or symptoms that can be treated with the pharmaceutical composition include inflammation, diseases caused by inflammation, obesity, metabolic syndrome, aging, and cancer.

[0062] The pharmaceutical composition can be suitably used for the prevention or treatment of such diseases or symptoms. In the present invention, treatment includes not only curing the disease or symptoms, but also alleviating or partially improving them. In the present invention, prevention includes inhibiting the progression of the disease or symptoms and inhibiting recurrence. In particular, the prevention or treatment of cancer includes reducing cancer tissue, preventing or inhibiting the progression of cancer, and suppressing the recurrence of cancer by inhibiting the proliferation of cancer cells.

[0063] In the present invention, the term "cancer" is interpreted in a broad sense and is used interchangeably with the term "malignant tumor." In addition, before a pathological diagnosis is confirmed, that is, before a tumor is confirmed as benign or malignant, it may collectively include benign tumors, benign / malignant borderline lesions, and malignant tumors. In general, cancers are named after the organ or tissue from which they developed, and the main types include tongue cancer, gum cancer, pharyngeal cancer, maxillary cancer, laryngeal 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, adrenal cancer, pituitary tumor, pineal tumor, uterine cancer, ovarian cancer, vaginal cancer, bladder cancer, kidney cancer, prostate cancer, urethral cancer, retinoblastoma, conjunctival cancer, neuroblastoma, glioma, glioblastoma, skin cancer, medulloblastoma, leukemia, malignant lymphoma, testicular tumor, osteosarcoma, rhabdomyosarcoma, leiomyosarcoma, angiosarcoma, liposarcoma, chondrosarcoma, Ewing's sarcoma, multiple myeloma, and acute myeloid leukemia. According to the characteristics of the site of the organ where the cancer originates, it is further classified into upper, middle and lower pharyngeal cancer, upper, middle and lower esophageal cancer, gastric cardia cancer, gastric pyloric cancer, cervical cancer, uterine cancer, etc., but these are not limiting and are included in the description of "cancer" in the present invention. The pharmaceutical composition of the present invention used in anticancer therapy (also called "anticancer pharmaceutical composition") is effective against "cancer" in general, but can be preferably used in particular against breast cancer and acute myeloid leukemia. In particular, among the compounds (I), compounds 1 and 4 and their pharma- ceutical acceptable salts and solvates have excellent anticancer effects, particularly in terms of the effect of inhibiting the proliferation of cancer cells in breast cancer and / or acute myeloid leukemia.

[0064] The term "anticancer pharmaceutical composition" refers to a pharmaceutical composition that exhibits a therapeutic or preventive effect against cancer, which is a target disease or pathology, and preferably refers to a pharmaceutical composition that exhibits a therapeutic effect. The therapeutic effect includes alleviating (alleviating) symptoms characteristic of cancer or associated symptoms, preventing or delaying the worsening of symptoms, and the like. The latter can be considered as a preventive effect in that it prevents the symptoms from becoming severe. Thus, the therapeutic effect and the preventive effect are partially overlapping concepts, and it is difficult to clearly distinguish them, and there is little practical benefit in doing so. A typical preventive effect is preventing or delaying the recurrence (onset) of symptoms characteristic of cancer. As long as the composition exhibits some therapeutic effect or preventive effect, or both, against cancer, it corresponds to an anticancer pharmaceutical composition. The therapeutic or preventive effect against cancer brought about by compound (I) etc. may also include improvement of complications of cancer (e.g., cachexia). In other words, in this specification, "anticancer" and "cancer treatment" may include not only the effect of inhibiting proliferation or shrinking the cancer tissue itself, but also the improvement of complications (preferably cachexia). The "anti-cancer pharmaceutical composition" of the present invention can also be used in combination with known anti-cancer agents or anti-cancer agents that will be developed in the future.

[0065] The pharmaceutical composition may consist of only the above-mentioned compound (I), or may be prepared by combining it with other pharmacopoeially acceptable ingredients and preparing it in a form suitable for the desired use such as the administration route and administration method by a known method. The pharmaceutical composition can be prepared according to a conventional method except for blending the active ingredient, compound (I), etc. When formulated into a pharmaceutical formulation, other pharmacopoeially acceptable ingredients (e.g., carriers, excipients, disintegrants, buffers, emulsifiers, suspending agents, soothing agents, stabilizers, preservatives, antiseptics, surfactants, lubricants, diluents, coating agents, sugar-coating agents, flavorings, emulsifying / solubilizing / dispersing agents, pH adjusters, isotonic agents, solubilizing agents, fragrances, coloring agents, dissolution aids, physiological saline, etc.) may be contained. The dosage form of the formulation is not particularly limited, and includes tablets, powders, fine granules, granules, capsules, syrups, liquids, suspensions, emulsions, jellies, injections, external preparations, inhalants, nasal drops, eye drops, and suppositories.

[0066] The pharmaceutical composition contains, as an active ingredient, the compound (I) in an amount necessary for obtaining the expected preventive or therapeutic effect (i.e., a therapeutically effective amount). The amount of the compound (I) in the pharmaceutical composition varies depending on the dosage form, but can be set within the range of, for example, about 0.01% by mass to about 99.9% by mass so as to achieve the desired dosage.

[0067] The pharmaceutical composition is administered to a subject by oral or parenteral administration (intravenous, intraarterial, subcutaneous, intradermal, intramuscular, or intraperitoneal injection, transdermal, nasal, transmucosal, etc.) depending on the dosage form. These administration routes are not mutually exclusive, and any two or more selected may be used in combination (for example, intravenous injection at the same time as oral administration or after a certain time has elapsed). Local administration may be used instead of systemic administration. The active ingredient may be administered so as to be delivered to a target tissue specifically using a drug delivery system (DDS). The "subject" here is not particularly limited, and includes humans and non-human mammals (including pet animals, livestock, and experimental animals, specifically, for example, mice, rats, guinea pigs, hamsters, monkeys, cows, pigs, goats, sheep, dogs, cats, chickens, quails, etc.) that require treatment or prevention. Preferably, the subject is a human.

[0068] The dosage of the pharmaceutical composition may generally vary depending on the symptoms, age, sex, and weight of the patient, but a person skilled in the art can set an appropriate dosage. For example, for oral administration, for example, about 0.01 mg to 1000 mg can be administered once or several times a day to an adult, but this is not limited thereto. In addition, for parenteral administration, for example, about 0.01 mg to 1000 mg can be administered once 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, etc. can be adopted. In setting the administration schedule, the symptoms of the patient and the duration of effect of the active ingredient can be taken into consideration.

[0069] (III) Food and drink composition The food and beverage composition of the present invention (hereinafter referred to as "the food and beverage composition") is characterized by containing the aforementioned GLS1 inhibitor, specifically, compound (I), a pharma- ceutically acceptable salt thereof, or a solvate thereof. The present food and drink composition can be prepared by, but is not limited to, adding the compound (I) in the manufacturing process of a conventionally known food and drink, or by adding the compound (I) to a conventionally known food and drink. The present food and drink composition can be effectively used, without being limited thereto, for example, in anti-obesity and prevention or amelioration of metabolic syndrome.

[0070] (IV) Cosmetic compositions The cosmetic composition according to the present invention (hereinafter referred to as "the present cosmetic composition") is characterized by containing the above-mentioned GLS1 inhibitor, specifically, the above-mentioned compound (I), its pharma- ceutically acceptable salt, or a solvate thereof. The present cosmetic composition also includes medicated cosmetics that fall under the category of quasi-drugs. The cosmetic composition of the present invention can be prepared by, but is not limited to, blending the compound (I) and the like in the manufacturing process of conventionally known cosmetics (including medicated cosmetics).The cosmetic composition can also be prepared by adding the compound (I) to conventionally known cosmetic ingredients. The present cosmetic composition is useful, for example but not limited to, for anti-aging, specifically for improving wrinkles and preventing aging.

[0071] As described above, in this specification, the terms "comprise" and "contain" include the meanings of "consist of" and "substantially consist of." Furthermore, the present invention is not limited to the above-described embodiments, and can be freely modified within the scope of the present invention. EXAMPLES

[0072] In the following, the present invention will be described using experimental examples to aid in understanding the configuration and effects of the present invention. However, the present invention is not limited by these experimental examples. The following experiments were carried out at room temperature (25±5°C) and atmospheric pressure conditions unless otherwise specified. In the following, "%" means "% by mass" and "parts" means "parts by mass" unless otherwise specified.

[0073] Except for the compounds prepared by the methods described below, chemical substances were purchased from Sigma-Aldrich, Merck (Darmstadt, Germany), Fujifilm Wako Chemicals (Osaka, Japan), Nacalai Tesque, Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan), and Kanto Chemical Co., Ltd. (Tokyo, Japan) and used as received. Silica gel 60N (spherical, neutral, particle size 63-210 nm, Kanto Chemical) was used for column chromatography, and Merck silica gel 60F was used for thin-layer chromatography. 254 Melting points were measured using a micro melting point apparatus (Yanaco) and are uncorrected. Nuclear magnetic resonance (NMR) spectra were measured using a JEOL JNM-A400 ( 1 H and 13 C, 400MHz and 100MHz, respectively) or JEOL JNM-ECX500 ( 1 H and 13The IR spectra were obtained at 500 MHz and 125 MHz for C, respectively. Chemical shifts (δ) are reported in ppm downfield from TMS, and coupling constants (J) are expressed in Hertz. IR spectra were measured using a JASCO FT / IR-460 Plus spectrophotometer (JASCO Corporation, Tokyo, Japan). High-resolution mass spectra were obtained on a Shimadzu LCMS-9090 equipped with a Dual Ion Source probe performing both electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI) sequentially.

[0074] Production Example 1: Synthesis of GLS1 inhibitor candidate compounds (compounds 1-18) Candidate GLS1 inhibitor compounds (compounds 1 to 18) were produced according to the procedure shown in the following scheme.

[0075] [ka]

[0076] (1) Step 1: Synthesis of amine compound 1) Under Ar atmosphere, bromoacetic acid (1.2 eq.) and potassium hydroxide (2 eq.) were added in order to an ethanol solution of 2-amino-5-mercapto-1,3,4-thiadiazole (1 eq.) under ice cooling, and the mixture was stirred at room temperature for 4 hours. After the reaction was completed, the solvent was removed using a rotary evaporator.

[0077] 2) The pH of the resulting residue was adjusted to 7 by adding 10% HCl (aq.). The precipitated solid was washed with dichloromethane and water to obtain a crude carboxylic acid. The resulting crude carboxylic acid was used in the next reaction without purification. Under an Ar atmosphere, triethylamine (1.5 eq.), 1-hydroxybenzotriazole monohydrate (1.5 eq.), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (1.5 eq.) and benzylamine (1.5 eq.) were added successively at room temperature to a dichloromethane solution of the resulting crude carboxylic acid (1 eq.), and the mixture was stirred at room temperature for 19 hours. After the reaction was completed, the solvent was removed using a rotary evaporator, and the resulting residue was purified by silica gel column chromatography (dichloromethane:methanol = 20:1) to obtain the desired amine compound 1.

[0078] (2) Step 2-A: Preparation of Compounds 1 to 4, 6 to 11, and 13 Under Ar atmosphere, triethylamine (1.5 eq.), 1-hydroxybenzotriazole monohydrate (1.5 eq.), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (1.5 eq.) and the corresponding carboxylic acid (1.5 eq.) were added to a dichloromethane solution of the amine compound (1 eq.) in sequence at room temperature, and the mixture was stirred at room temperature for 15 hours. After the reaction was completed, the solvent was removed using a rotary evaporator, and the resulting residue was purified by silica gel column chromatography (n-hexane: ethyl acetate = 30: 1 to 10: 1) to obtain the desired compounds 1 to 4, 6 to 11, and compound a.

[0079] To a stirred solution of compound a (36 mg, 0.08 mmol, 1.0 eq) in methanol (1.5 mL) was added 10% aqueous hydrochloric acid (2 drops) at room temperature, and the resulting mixture was heated at 50° C. for 18 h. The solvent was evaporated to give a residue, which was purified by silica gel column chromatography (dichloromethane:methanol=10:1) to give compound 13 (32 mg, 0.08 mmol, 100%) as a white solid.

[0080] (3) Step 2-B: Preparation of Compound 12 Under Ar atmosphere, triethylamine (1.5 eq.), 4-dimethylaminopyridine (1.5 eq.), N,N-dicyclohexylcarbodiimide (1.5 eq.) and the corresponding carboxylic acid (1.5 eq.) were added to a dichloromethane solution of the amine compound (1 eq.) in sequence at room temperature, and the mixture was stirred at room temperature for 15 hours. After the reaction was completed, the solvent was removed using a rotary evaporator, and the resulting residue was purified by silica gel column chromatography (n-hexane: ethyl acetate = 10:1 to 1:1) to obtain the desired compound 12.

[0081] (4) Step 2-C: Preparation of Compounds 5 and 14-18 Under an Ar atmosphere, triethylamine (1.5 eq.) was added to an acetonitrile solution of an amine compound (1 eq.) at room temperature, and then the corresponding acyl chloride (1.5 eq.) was added under ice cooling. The reaction solution was then stirred at 80°C for 3 hours. After the reaction was completed, the solvent was removed using a rotary evaporator, and the resulting residue was purified by silica gel column chromatography (dichloromethane:methanol = 80:1 to 10:1) to obtain the desired compounds 5 and 14-18.

[0082] Production Example 2: Synthesis of GLS1 inhibitor candidate compounds (compounds 19 to 24) Candidate GLS1 inhibitor compounds (compounds 19 to 24) were produced according to the procedure shown in the formula below.

[0083] [ka]

[0084] 1) Under an Ar atmosphere, bromoacetic acid (1.2 eq.) and potassium hydroxide (2 eq.) were added successively to an ethanol solution of 2-amino-5-mercapto-1,3,4-thiadiazole (1 eq.) under ice cooling, and the mixture was stirred at room temperature for 4 hours. After the reaction was completed, the solvent was removed using a rotary evaporator, and the pH of the resulting residue was adjusted to 7 by adding 10% HCl (aq.). The precipitated solid was washed with dichloromethane and water to obtain a crude carboxylic acid. The crude carboxylic acid obtained was used in the next reaction without purification.

[0085] 2) Under an Ar atmosphere, triethylamine (1.5 eq.), 1-hydroxybenzotriazole monohydrate (1.5 eq.), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (1.5 eq.), and the corresponding benzylamine (1.5 eq.) were added successively to a dichloromethane solution of the obtained crude carboxylic acid (1 eq.) at room temperature, and the mixture was stirred at room temperature for 19 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 (dichloromethane:methanol=20:1) to obtain the desired amine compounds 2-4.

[0086] 3) Under an Ar atmosphere, triethylamine (1.5 eq.) was added to an acetonitrile solution of the amine compounds 2-4 (1 eq.) at room temperature, and then the corresponding acyl chloride (1.5 eq.) was added under ice cooling. The reaction solution was then stirred at 80°C for 3 hours. After the reaction was completed, the solvent was removed using a rotary evaporator, and the resulting residue was purified by silica gel column chromatography (dichloromethane:methanol = 80:1 to 10:1) to obtain the desired compounds 19-24.

[0087] The characteristics of each compound synthesized by the above mentioned method are shown below. Amine compound 1: 2-((5-amino-1,3,4-thiadiazol-2-yl)thio)-N-benzylacetamide Yield: 38% in 2 steps; 1 H-NMR (DMSO-d6, 400 MHz) δ: 8.65 (1H, t, J = 5.2 Hz), 7.32-7.28 (5H, m), 7.24-7.20 (2H, m), 4.27 (2H, d, J= 6.0 Hz), 3.83 (2H, s).

[0088] Amine compound 2: 2-((5-amino-1,3,4-thiadiazol-2-yl)thio)-N-(3-methoxybenzyl)acetamide Yield: 61% in 2 steps; 1 H-NMR (DMSO-d6, 400 MHz) δ: 8.65 (1H, t, J = 6.0 Hz), 7.30 (2H, s), 7.21 (1H, t, J = 6.4 Hz), 6.85-6.75 (3H, m), 4.26 (2H, d, J = 6.0 Hz), 3.84 (2H, s), 3.73 (3H, s); 13 C-NMR (DMSO-d6, 100 MHz) δ: 169.94, 167.03, 159.34, 149.64, 140.58, 129.41, 119.36, 112.66, 112.46, 55.02, 42.48, 37.70; IR (KBr): 3381, 3188, 3082, 3055, 1637, 1518 cm -1 ; HRMS (ESI): Calcd for C 12 H 14 N4O2S2311.0631 ([M+H] + ), Found 311.0620.

[0089] Amine compound 3: 2-((5-amino-1,3,4-thiadiazol-2-yl)thio)-N-(4-methoxybenzyl)acetamide Yield: 41% in 2 steps; 11H-NMR (DMSO-d6, 400 MHz) δ: 8.57 (1H, t, J = 6.0 Hz), 7.29 (2H, s), 7.12 (2H, d, J = 8.0 Hz), 6.85 (2H, d, J = 8.0 Hz), 4.18 (2H, d, J= 6.0 Hz), 3.70 (3H, s); 13 13C-NMR (DMSO-d6, 125 MHz) δ: 169.65, 166.48, 157.95, 149.25, 130.53, 128.27, 113.39, 54.76, 41.99, 37.78; IR (KBr): 3331, 3314, 3288, 3103, 3069, 3013, 2959, 1645, 1520, 1246 cm -1 ; HRMS (ESI): Calcd for C 12 H 14 N4O2S2 311.0631 ([M+H] + ), Found 311.0621.

[0090] Amine compound 4: 2-((5-amino-1,3,4-thiadiazol-2-yl)thio)-N-(3,4-dimethoxybenzyl)acetamide Yield: 54% in 2 steps; 1 1H-NMR (DMSO-d6, 400 MHz) δ: 8.57 (1H, t, J = 6.0 Hz), 7.30 (2H, s), 6.87 (1H, d, J = 8.0 Hz), 6.84 (1H, d, J = 2.0 Hz), 6.74 (1H, dd, J = 8.0, 2.0 Hz), 4.20 (2H, d, J = 6.0 Hz), 3.82 (2H, s), 3.71 (3H, s), 3.70 (3H, s); 13 13C-NMR (DMSO-d6, 100 MHz) δ: 169.95, 166.88, 149.66, 148.67, 147.81, 131.32, 119.35, 111.68, 111.18, 55.59, 55.43, 54.97, 42.32, 37.74; IR (KBr): 3383, 3358, 3219, 3182, 1628, 1522, 1418, 1261, 1030 cm -1 ; HRMS (ESI): Calcd for C13H16N4O3S2 341.0737 ([M+H]+), Found 341.0726.

[0091] Compound 1: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)cyclobutanecarboxamide Yield: 97% 1 1H-NMR (DMSO-d6, 400 MHz) δ: 12.49 (1H, s), 8.73 (1H, t, J = 5.6 Hz), 7.30 - 7.20 (5H, m), 4.29 (2H, d, J = 5.6 Hz), 4.01 (2H, s), 3.37 (1H, quin, J = 8.0 Hz), 2.26 - 2.10 (4H, m), 2.01 - 1.89 (1H, m), 1.86 - 1.76 (1H, m); 13 13C-NMR (DMSO-d6, 125 MHz) δ: 173.65, 167.22, 159.69, 158.54, 139.43, 128.78, 127.72, 127.36, 43.06, 38.66, 37.58, 25.01, 18.28; mp: 209 - 211 ℃; IR (KBr): 2986, 2889, 2781, 1688, 1641, 1564, 1541, 1296, 696 cm -1 ; HRMS (ESI): Calcd for C 16 H 18 N4O2S2 363.0944 ([M+H] + ), Found 363.0936。

[0092] Compound 2: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)cyclopentanecarboxamide Yield: 87% 11H-NMR (CDCl3, 400 MHz) δ: 11.83 (1H, s), 7.25 - 7.17 (5H, m), 4.45 (2H, d, J = 6.0 Hz), 3.96 (2H, s), 2.98 (1H, quin, J = 7.2 Hz), 2.02 - 1.94 (2H, m), 1.90 - 1.83 (2H, m), 1.81 - 1.72 (2H, m), 1.70 - 1.61 (2H, m); 13 13C-NMR (CDCl3, 100 MHz) δ: 175.17, 167.27, 161.28, 159.01, 137.73, 128.77, 127.66, 127.54, 45.26, 44.00, 36.27, 30.37, 26.22; mp: 201 - 202 ℃; IR (KBr): 3140, 2995, 1653, 1543, 1028 cm -1 ; HRMS (ESI): Calcd for C 17 H 20 N4O2S2 377.1100 ([M + H] + ), Found 377.1096。

[0093] Compound 3: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)cyclohexanecarboxamide Yield: 78% 1 1H-NMR (CDCl3, 400 MHz) δ: 12.11 (1H, s), 7.23 - 7.16 (5H, m), 4.45 (2H, d, J = 6.0 Hz), 3.99 (2H, s), 2.55 (1H, tt, J = 12.0, 3.2 Hz), 1.89 (2H, d, J = 13.2 Hz), 1.81 (2H, dt, J = 13.2, 3.2 Hz), 1.72 (1H, d, J = 12.0 Hz), 1.52 (2H, qd, J = 10.0, 2.4 Hz), 1.40 - 1.19 (3H, m); 1313C-NMR (CDCl3, 100 MHz) δ: 174.89, 167.22, 161.26, 159.16, 137.69, 128.77, 127.67, 127.64, 44.65, 44.10, 36.09, 29.20, 25.65, 25.59; mp: 213 - 214 °C; IR (KBr): 3306, 3292, 3245, 1682, 1639, 1508, 1398, 1169, 750 cm -1 ; HRMS (ESI): Calcd for C 18 H 22 N4O2S2 391.1257 ([M + H] + ), Found 391.1252。

[0094] Compound 4: trans-N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-hydroxycyclohexanecarboxamide Yield: 60% 1 1H-NMR (Methanol-d4, 400 MHz) δ: 7.29 - 7.21 (5H, m), 4.38 (2H, s), 3.97 (2H, s), 3.55 (1H, tt, J = 13.2, 3.6 Hz), 2.45 (1H, tt, J = 13.2, 3.6 Hz), 2.04 (2H, dd, J = 13.2, 3.6 Hz), 1.96 (2H, d, J = 13.2 Hz), 1.60 (2H, qd, J = 13.2, 3.6 Hz), 1.33 (2H, qd, J = 13.2, 3.6 Hz); 13 13C-NMR (DMSO-d6, 100 MHz) δ: 174.27, 166.69, 159.03, 158.03, 138.91, 128.27, 127.20, 126.84, 68.05, 42.63, 42.54, 42.42, 37.09, 37.05, 34.46, 27.74; mp: 214 - 216 °C; IR (KBr): 3292, 2926, 2905, 1676, 1639, 1556, 1063, 741 cm -1 ; HRMS (ESI): Calcd 18 H 22 N4O3S2407.1206 ([M+H] + ), Found 407.1201.

[0095] Compound 5: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)isobutyramide Share: 73% 1 H-NMR (DMSO-d6, 400 MHz) δ: 8.72 (1H, t, J = 5.6 Hz), 7.30–7.20 (5H, m), 4.28 (2H, d, J = 5.6 Hz), 4.01 (2H, s), 2.76 (1H, Sept, J = 6.8 Hz), 1.11 (6H, d, J = 6.8 Hz); 13 C-NMR (DMSO-d6, 100 MHz) δ: 175.49, 166.71, 159.18, 158.09, 138.97, 128.30, 127.23, 126.87, 19.04; mp: 185-187 °C; IR (KBr): 3298, 3034, 2941, 2874, 1684, 1643, 1553, 1296, 1202, 698 cm -1 ; HRMS (ESI): Calcd 15 H 18 N4O2S2351.0944 ([M+H] + ), Found 351.0938.

[0096] Compound 6: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)hexanamide Rate: 95% 11H-NMR (CDCl3, 400 MHz) δ: 11.82 (1H, s), 7.25 - 7.18 (5H, m), 4.45 (2H, d, J = 6.0 Hz), 3.96 (2H, s), 2.50 (2H, t, J = 7.2 Hz), 1.70 (2H, quin, J = 7.2 Hz), 1.34 - 1.30 (4H, m), 0.89 (3H, t, J = 7.2 Hz); 13 13C-NMR (DMSO-d6, 100 MHz) δ: 171.70, 166.69, 159.08, 157.95, 138.94, 128.26, 127.20, 126.82, 42.54, 37.04, 34.73, 30.70, 24.29, 21.83, 13.82; mp: 228 - 230 °C; IR (KBr): 3306, 3267, 3248, 3200, 3163, 1684, 1645, 1539, 1508, 1456 cm -1 ; HRMS (ESI): Calcd for C 17 H 22 N4O2S2 379.1257 ([M + H] + ), Found 379.1255。

[0097] Compound 7: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-fluorobenzamide Yield: 28% 1 1H-NMR (DMSO-d6, 400 MHz) δ: 8.74 (1H, t, J = 6.0 Hz), 8.20 - 8.17 (2H, m), 7.40 (2H, t, J = 7.2 Hz), 7.31 - 7.19 (5H, m), 4.30 (2H, d, J = 6.0 Hz), 4.05 (2H, s); 1313C-NMR (DMSO-d6, 100 MHz) δ: 166.75, 166.04, 164.64, 162.27 (d, J = 254.5 Hz), 158.07, 138.95, 131.30 (d, J = 9.5 Hz), 128.66, 128.28, 127.23, 126.94, 115.67 (d, J = 22.0 Hz), 42.56, 37.03; mp: 213 - 214 °C; IR (KBr): 3260, 3092, 3063, 1647, 1543, 1506, 1315, 669 cm -1 ; HRMS (ESI): Calcd for C 18 H 15 FN4O2S2 403.0693 ([M + H] + ), Found 403.0678。

[0098] Compound 8: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-chlorobenzamide Yield: 36% 1 1H-NMR (DMSO-d6, 400 MHz) δ: 8.75 (1H, t, J = 6.0 Hz), 8.11 (2H, dd, J = 7.2, 2.4 Hz), 7.64 (2H, dd, J = 7.2, 2.4 Hz), 7.31 - 7.19 (5H, m), 4.30 (2H, d, J = 6.0 Hz), 4.06 (2H, s); 13 13C-NMR (DMSO-d6, 100 MHz) δ: 166.98, 164.67, 160.21, 159.00, 139.03, 138.22, 130.53, 130.38, 129.03, 128.48, 127.37, 127.06, 42.76, 37.20; mp: 281 - 282 °C; IR (KBr): 3152, 3047, 3015, 1636, 1533, 1456, 1313, 1097, 696 cm -1 ; HRMS (ESI): Calcd for C 18 H 15 ClN4O2S2 417.0252 ([M-H] - ), Found 417.0242。

[0099] Compound 9: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-methylbenzamide Yield: 17% 1 1H-NMR (DMSO-d6, 400 MHz) δ: 8.75 (1H, t, J = 5.6 Hz), 8.01 (2H, d, J = 8.4 Hz), 7.36 (2H, d, J = 8.4 Hz), 7.30 - 7.19 (5H, m), 4.30 (2H, d, J = 5.6 Hz), 4.05 (2H, s), 2.39 (3H, s); 13 13C-NMR (DMSO-d6, 125 MHz) δ: 166.72, 165.25, 160.50, 158.21, 143.32, 138.93, 129.22, 128.85, 128.45, 128.27, 127.22, 126.83, 42.55, 37.06, 21.14; mp: 230 - 232 °C; IR (KBr): 3292, 3167, 2943, 1649, 1541, 1292 cm -1 ; HRMS (ESI): Calcd for C 19 H 18 N4O2S2 399.0944 ([M+H] + ), Found 399.0933。

[0100] Compound 10: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-(trifluoromethyl)benzamide Yield: 18% 11H-NMR (DMSO-d6, 400 MHz) δ: 8.75 (1H, t, J = 6.0 Hz), 8.27 (2H, d, J= 8.0 Hz), 7.94 (2H, d, J = 8.0 Hz), 7.33 - 7.19 (5H, m), 4.30 (2H, d,J = 6.0 Hz), 4.06 (2H, s); 13 13C-NMR (DMSO-d6, 125 MHz) δ: 166.85, 165.19, 161.82, 157.92, 138.98, 136.72, 132.07 (d, J = 32.3 Hz), 129.34, 128.33, 127.26, 126.89, 125.52 (d, J = 14.5 Hz), 123.93 (q, J = 270.0 Hz), 42.61, 37.06; mp: 258 - 260 ℃; IR (KBr): 3323, 3290, 3223, 3194, 1663, 1638, 1543, 1333, 1123, 696 cm -1 ; HRMS (ESI): Calcd for C 19 H 15 F3N4O2S2453.0661 ([M + H] + ), Found 453.0651。

[0101] Compound 11: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-nitrobenzamide Yield: 17% 1 1H-NMR (DMSO-d6, 400 MHz) δ: 8.76 (1H, t, J = 6.0 Hz), 8.37 (2H, d, J= 8.8 Hz), 8.31 (2H, d, J = 8.8 Hz), 7.31 - 7.19 (5H, m), 4.30 (2H, d,J = 6.0 Hz), 4.06 (2H, s); 1313C-NMR (DMSO-d6, 125 MHz) δ: 166.76, 164.87, 157.85, 149.57, 138.95, 138.79, 129.89, 128.27, 127.22, 127.09, 126.82, 123.57, 42.54, 36.98; mp: 262 - 264 °C; IR (KBr): 3308, 3246, 3115, 2937, 1666, 1627, 1558, 1541, 1315, 694 cm -1 ; HRMS (ESI): Calcd for C 18 H 15 N5O4S2 428.0493 ([M - H] - ), Found 428.0483。

[0102] Compound 12: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-methoxybenzamide Yield: 15% 1 1H-NMR (DMSO-d6, 400 MHz) δ: 8.74 (1H, t, J = 6.0 Hz), 8.10 (2H, d, J = 9.2 Hz), 7.30 - 7.19 (5H, m), 7.05 (2H, d, J = 9.2 Hz), 4.30 (2H, d, J = 6.0 Hz), 4.02 (2H, s), 3.84 (3H, s); 13 13C-NMR (DMSO-d6, 100 MHz) δ: 166.77, 164.70, 163.01, 160.65, 158.09, 138.97, 130.57, 128.31, 127.25, 126.87, 123.68, 114.01, 55.61, 42.57, 37.06; mp: 248 - 249 °C; IR (KBr): 3323, 3182, 2997, 2845, 2673, 1641, 1607, 1558, 1296, 1178, 690 cm -1 ; HRMS (ESI): Calcd for C 19H 18 N4O3S2415.0893 ([M+H] + ), Found 415.0873。

[0103] Compound a: N-(4-(Methoxymethoxy)benzyl)-2-((5-(2-phenylacetamido)-1,3,4-thiadiazol-2-yl)thio)acetamide Yield: 17%; 1 1H-NMR (DMSO-d6, 400 MHz) δ: 8.76 (1H, t, J = 6.0 Hz), 8.11 (2H, d, J = 9.2 Hz), 7.33 - 7.20 (5H, m), 7.16 (2H, d, J = 9.2 Hz), 5.31 (2H, s), 4.31 (2H, d, J = 6.0 Hz), 4.06 (2H, s), 3.40 (3H, s); 13 13C-NMR (DMSO-d6, 100 MHz) δ: 166.71, 164.43, 160.52, 160.16, 158.43, 138.95, 130.51, 128.30, 127.24, 126.86, 124.31, 115.84, 93.67, 55.90, 42.56, 37.05; mp: 210 - 212 ℃; IR (KBr): 3373, 3302, 3352, 3271, 1059 cm -1 ; HRMS (ESI): Calcd for C 20 H 20 N4O4S2445.0999 ([M+H] + ), Found 445.0983。

[0104] Compound 13: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-hydroxybenzamide Yield: 17% 11H-NMR (DMSO-d6, 400 MHz) δ: 10.42 (1H, s), 8.75 (1H, t, J = 5.6 Hz), 8.01 (2H, d, J = 8.8 Hz), 7.30 - 7.19 (5H, m), 6.88 (2H, d, J = 8.8 Hz), 4.30 (2H, d, J = 5.6 Hz), 4.04 (2H, s); 13 13C-NMR (DMSO-d6, 100 MHz) δ: 166.75, 162.10, 158.32, 138.97, 130.78, 128.96, 128.65, 128.31, 127.25, 126.87, 121.59, 115.40, 42.58, 37.08; mp: 249 - 250 °C; IR (KBr): 3778, 3676, 3382, 3227, 3211, 1609, 1587, 1539, 1285 cm -1 ; HRMS (ESI): Calcd for C 18 H 16 N4O3S2 401.0737 ([M + H] + ), Found 401.0723。

[0105] Compound 14: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-(trifluoromethoxy)benzamide Yield: 100% 1 1H-NMR (DMSO-d6, 400 MHz) δ: 8.76 (1H, t, J = 5.6 Hz), 8.23 (2H, d, J = 8.4 Hz), 7.56 (2H, d, J = 8.4 Hz), 7.31 - 7.19 (5H, m), 4.30 (2H, d, J = 5.6 Hz), 4.06 (2H, s); 1313C-NMR (DMSO-d6, 100 MHz) δ: 166.71, 164.20, 159.95, 158.91, 151.50, 138.97, 131.02, 130.52, 128.32, 127.26, 126.89, 120.86, 119.97 (q, J = 255.5 Hz), 42.60, 37.05; mp: 190 - 192 °C; IR (KBr): 3356, 3173, 1692, 1261, 1022 cm -1 ; HRMS (ESI): Calcd for C 19 H 15 F3N4O3S2 469.0610 ([M + H] + ), Found 469.0595。

[0106] Compound 15: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-2-(trifluoromethyl)benzamide Yield: 44% 1 1H-NMR (DMSO-d6, 400 MHz) δ: 8.75 (1H, t, J = 5.6 Hz), 7.89 (1H, d, J = 7.2 Hz), 7.79 (3H, m), 7.31 - 7.20 (5H, m), 4.30 (2H, d, J = 5.6 Hz), 4.06 (2H, s); 13 13C-NMR (DMSO-d6, 100 MHz) δ: 166.99, 165.99, 159.42, 159.12, 139.00, 132.87, 131.45, 129.23, 128.47, 127.34, 127.04, 126.81, 126.77, 126.50 (d, J = 31.5 Hz), 123.71 (q, J = 272.6 Hz), 42.77, 37.21; mp: 198 - 199 °C; IR (KBr): 3503, 3018, 2986, 2939, 1651, 1437, 1308, 1022 cm -1 ; HRMS (ESI): Calcd for C19 H 15 F3N4O2S2453.0661 ([M+H] + ), Found 453.0645。

[0107] Compound 16: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-3-(trifluoromethyl)benzamide Yield: 77% 1 H-NMR (DMSO-d6, 400 MHz) δ: 8.77 (1H, t, J = 6.0 Hz), 8.48 (1H, s), 8.36 (1H, d, J = 7.6 Hz), 8.04 (1H, d, J = 7.6 Hz), 7.81 (1H, t, J= 7.6 Hz), 7.31 - 7.18 (5H, m), 4.30 (2H, d, J = 6.0 Hz), 4.07 (2H, s); 13 C-NMR (DMSO-d6, 100 MHz) δ: 166.72, 164.12, 159.99, 159.08, 138.96, 132.62, 130.09, 129.61, 129.53, 129.29, 128.33, 127.27, 126.89, 125.14, 123.85 (q, J = 270.7 Hz), 42.62, 37.04; mp: 239 - 240 ℃; IR (KBr): 3404, 3375, 3283, 1647, 1541, 1337, 1132 cm -1 ; HRMS (ESI): Calcd for C 19 H 15 F3N4O2S2453.0661 ([M+H] + ), Found 453.0645。

[0108] Compound 17: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-2-fluorobenzamide Yield: 49% 1H-NMR (DMSO-d6, 400 MHz) δ: 8.75 (1H, t, J = 6.0 Hz), 7.76 (1H, td, J = 7.6, 2.0 Hz), 7.65 (1H, qd, J = 7.6, 2.0 Hz), 7.41-7.34 (2H, m), 7.31-7.20 (5H, m), 4.30 (2H, d, J = 6.0 Hz), 4.06 (2H, s); 13 C-NMR (DMSO-d6, 125 MHz) δ: 166.67, 162.90, 159.49 (d, J = 251.6 Hz), 159.06, 158.93, 138.93, 134.16 (d, J = 8.4 Hz), 130.47, 128.28, 127.24, 126.85, 124.68, 121.55 (d, J = 13.1 Hz), 116.46 (d, J = 20.3 Hz), 42.58, 37.12; mp: 179-180 ℃; IR (KBr): 3474, 3456, 3221, 3207, 1684, 1520, 1456, 1306 cm -1 ; HRMS (ESI): Calcd for C 18 H 15 FN4O2S2403.0693 ([M+H] + ), Found 403.0676。

[0109] Compound 18: N-(5-((2-(Benzylamino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-3-fluorobenzamide Yield: 66% 1 H-NMR (DMSO-d6, 400 MHz) δ: 8.76 (1H, t, J = 6.4 Hz), 7.94 (2H, t, J= 7.6 Hz), 7.65-7.59 (1H, m), 7.53 (1H, td, J = 8.8, 2.4 Hz), 7.33-7.19 (5H, m), 4.30 (2H, d, J = 6.4 Hz), 4.07 (2H, s); 1313C-NMR (DMSO-d6, 100 MHz) δ: 166.68, 163.95, 161.94 (d, J = 243.1 Hz), 159.76, 158.96, 138.94, 133.60, 130.97 (d, J = 7.6 Hz), 128.30, 127.25, 126.87, 124.70, 120.06 (d, J = 20.9 Hz), 115.24 (d, J = 23.8 Hz), 42.59, 37.05; mp: 243 - 244 °C; IR (KBr): 3298, 3283, 3171, 2939, 1647, 1558, 1456, 1304 cm -1 ; HRMS (ESI): Calcd for C 18 H 15 FN4O2S2 403.0693 ([M + H] + ), Found 403.0676。

[0110] Compound 19: N-(5-((2-((3-Methoxybenzyl)amino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)isobutyramide Yield: 100% 1 1H-NMR (DMSO-d6, 400 MHz) δ: 8.70 (1H, t, J = 5.6 Hz), 7.19 (1H, t, J = 7.6 Hz), 6.79 - 6.78 (2H, m), 6.78 (1H, s), 4.26 (2H, d, J = 5.6 Hz), 4.01 (2H, s), 3.71 (3H, s), 2.75 (1H, sept, J = 6.8 Hz), 1.11 (6H, d, J = 6.8 Hz); 13 13C-NMR (DMSO-d6, 100 MHz) δ: 175.47, 166.74, 159.32, 159.15, 158.12, 140.55, 129.36, 119.36, 112.71, 112.42, 55.00, 42.52, 37.05, 33.83, 19.02; mp: 178 - 179 °C; IR (KBr): 3396, 3182, 3140, 2989, 2903, 1691, 1645, 1547, 1292, 1028 cm -1 ; HRMS (ESI): Calcd for C 16 H 20 N4O3S2 381.1050 ([M+H] + ), Found 381.1053。

[0111] Compound 20: N-(5-((2-((4-Methoxybenzyl)amino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)isobutyramide Yield: 90% 1 1H-NMR (DMSO-d6, 400 MHz) δ: 8.64 (1H, t, J = 6.0 Hz), 7.13 (2H, d, J = 8.4 Hz), 6.84 (2H, d, J = 8.4 Hz), 4.21 (2H, d, J = 6.0 Hz), 3.97 (2H, s), 3.71 (3H, s), 2.75 (1H, sept, J = 6.8 Hz), 1.11 (6H, d, J = 6.8 Hz); 13 13C-NMR (DMSO-d6, 100 MHz) δ: 175.49, 166.54, 159.19, 158.27, 158.11, 130.85, 128.60, 113.68, 55.08, 42.04, 37.15, 33.84, 19.02; mp: 186 - 188 ℃; IR (KBr): 3466, 3389, 3275, 2974, 1684, 1630, 1516, 1259 cm -1 ; HRMS (ESI): Calcd for C 16 H 20 N4O3S2 381.1050 ([M+H] + ), Found 381.1036。

[0112] Compound 21: N-(5-((2-((3,4-dimethoxybenzyl)amino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)isobutyramide [N-(5-((2-((3,4-Dimethoxybenzyl)amino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)isobutyramide] Rate: 76% 1 H-NMR (DMSO-d6, 400 MHz) δ: 8.62 (1H, t, J = 6.0 Hz), 6.84 (1H, d, J = 8.0 Hz), 6.82 (1H, d, J = 1.6 Hz), 6.73 (1H, dd, J = 8.0, 1.6 Hz), 4.20 (2H, d, J = 6.0 Hz), 3.98 (2H, s), 3.70 (3H, s), 3.69 (3H, s), 2.75 (1H, Sept, J = 6.8 Hz), 1.10 (6H, d, J = 6.8 Hz); 13 C-NMR (DMSO-d6, 100 MHz) δ: 175.51; 42.38, 37.10, 33.84, 19.02; mp: 194-195 °C; IR (KBr): 3481, 3406, 3327, 3186, 3092, 2966, 1641, 1522, 1296, 1273 cm -1 ; HRMS (ESI): Calcd 17 H 22 N4O4S2411.1155 ([M+H] + ), Found 411.1141.

[0113] Compound 22: N-(5-((2-((3-Methoxybenzyl)amino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-(trifluoromethyl)benzamide Rate: 91% 11H-NMR (DMSO-d6, 400 MHz) δ: 8.74 (1H, t, J = 6.0 Hz), 8.27 (2H, d, J= 8.4 Hz), 7.94 (2H, d, J = 8.4 Hz), 7.20 (1H, t, J = 8.0 Hz), 6.82-6.76 (3H, m), 4.27 (2H, d, J = 6.0 Hz), 4.07 (2H, s), 3.70 (3H, s); 13 13C-NMR (DMSO-d6, 100 MHz) δ: 166.90, 164.60, 160.10, 159.44, 159.16, 140.62, 135.47, 132.81, 132.48, 129.53, 125.80 (d, J = 2.8 Hz), 123.90 (q, J = 271.7 Hz), 119.50, 112.86, 112.49, 55.11, 42.67, 37.11; mp: 185-186 °C; IR (KBr): 3396, 3244, 3060, 2945, 1661, 1547, 1329, 1026 cm -1 ; HRMS (ESI): Calcd for C 20 H 17 F3N4O3S2483.0767 ([M+H] + ), Found 483.0750。

[0114] Compound 23: N-(5-((2-((4-Methoxybenzyl)amino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-(trifluoromethyl)benzamide Yield: 63% 1 1H-NMR (DMSO-d6, 400 MHz) δ: 8.68 (1H, t, J = 5.6 Hz), 8.27 (2H, d, J= 8.0 Hz), 7.94 (2H, d, J = 8.0 Hz), 7.15 (2H, d, J = 8.8 Hz), 6.84 (2H, d, J = 8.8 Hz), 4.22 (2H, d, J = 5.6 Hz), 4.04 (2H, s), 3.67 (3H, s); 1313C-NMR (DMSO-d6, 100 MHz) δ: 166.79, 164.74, 160.22, 159.18, 158.43, 135.51, 132.72 (q, J = 31.5 Hz), 130.98, 129.57, 128.79, 125.85 (d, J = 2.8 Hz), 123.94 (q, J = 270.8 Hz), 113.85, 55.20, 42.25, 37.25; mp: 210 - 212 °C; IR (KBr): 3373, 3086, 2951, 1636, 1516, 1329, 1117 cm -1 ; HRMS (ESI): Calcd for C 20 H 17 F3N4O3S2 483.0767 ([M+H] + ), Found 483.0771。

[0115] Compound 24: N-(5-((2-((3,4-dimethoxybenzyl)amino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-(trifluoromethyl)benzamide [ N-(5-((2-((3,4-Dimethoxybenzyl)amino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)-4-(trifluoromethyl)benzamide] Yield: 47% 1 1H-NMR (DMSO-d6, 400 MHz) δ: 8.67 (1H, t, J = 5.6 Hz), 8.28 (2H, d, J = 8.4 Hz), 7.95 (2H, d, J = 8.4 Hz), 6.85 (1H, d, J = 8.4 Hz), 6.84 (1H, d, J = 1.6 Hz), 6.76 (1H, dd, J = 8.4, 1.6 Hz), 4.23 (2H, d, J = 5.6 Hz), 4.05 (2H, s), 3.71 (3H, s), 3.68 (3H, s); 13C-NMR (DMSO-d6, 100 MHz) δ: 166.75, 162.10, 158.32, 138.97, 130.78, 128.96, 128.65, 128.31, 127.25, 126.87, 121.59, 115.40, 42.58, 37.08; MP: 249-250℃; IR (KBr): 3778, 3676, 3382, 3227, 3211, 1609, 1587, 1539, 1285 cm -1 ; HRMS (ESI): Calcd for C 18 H 16 N4O3S2401.0737 ([M+H] + ), Found 401.0723.

[0116] Experimental Example 1. Experimental Materials (1) Reagents 6-Diazo-5-oxo-L-norleucine (DON) (Wako Pure Chemical Industries, Ltd.), Compound A: 5-amino-N-(benzo[d][1,3]dioxol-5-ylmethyl)-1-(2-oxo-2-(o-tolylamino)ethyl)-1H-1,2,3-triazole-4-carboxamide (F-1601-0438:Life Chemicals) Compound B: 5-amino-N-(benzo[d][1,3]dioxol-5-ylmethyl)-1-(2-((3,4-dimethylphenyl)amino)-2-oxoethyl)-1H-1,2,3-triazole-4-carboxamide (F-1602-0566:Life Chemicals) Compound C: N-(benzo[d][1,3]dioxol-5-ylmethyl)-2-((5-(2-phenylacetamido)-1,3,4-thiadiazol-2-yl)thio)acetamide (F-1838-2412:Life Chemicals) Compound D: N-(benzo[d][1,3]dioxol-5-ylmethyl)-2-((5-((2,3-dimethylphenoxy)methyl)-4-methyl-4H-1,2,4-triazol-3-yl)thio)acetamide (ASN 03799252: ASINEX) Compound E: N-(5-((2-((benzo[d][1,3]dioxol-5-ylmethyl)amino)-2-oxoethyl)thio)-1,3,4-thiadiazol-2-yl)cyclobutanecarboxamide (ASN 06400834:ASINEX)

[0117] 2. Generation of human recombinant GLS1 DNA encoding the amino acid sequence of hGLS1 (residues 73-669) was inserted into the NdeI and BamHI sites of pOPTH plasmid (Obita et al., 2007), and the resulting plasmid was used to transform E. coli (SoruBL21 strain). When the OD600nm of the culture reached 0.6, 0.5 mM IPTG was added to induce the expression of hGLS1. After overnight culture at 37°C, the cells were collected by centrifugation at 4°C and resuspended in a buffer containing 20 mM Tris-HCl (pH 8.0, 4°C), 500 mM NaCl, 10% glycerol, and 5 mM β-mercaptoethanol. The cells were lysed by sonication on ice, and then centrifuged at 10,200×g for 60 min at 4°C to obtain the supernatant. The resulting His-tagged hGLS1 protein was purified using a Ni-NTA column, and then by gel filtration chromatography using a Superdex HiLoad16 / 60 column equilibrated with a buffer containing 20 mM Tris (pH 8.0, 4°C) and 100 mM NaCl. The purified protein is composed of hGLS1 (residues 73-669) and an N-terminal His tag (AHHHHHM: SEQ ID NO: 3) (Non-Patent Documents 7 and 8).

[0118] 3. Evaluation of GLS1 inhibitory activity (1) Evaluation of the inhibitory effect of human recombinant GLS1 on Glu production 2.5 μL of a GLS1 inhibitor candidate compound (test compound) dissolved in a solvent (DMSO) or 2.5 μL of the solvent (DMSO) alone (for control) was added to 10.5 μL of a reaction solution (Tris-acetate (50 mM, pH 8.6), KPO4 (150 mM, pH 8.0), EDTA (0.2 mM, pH 8.0)), which was mixed with a recombinant GLS1 stock solution and incubated at 37°C for 20 minutes. L-glutamine solution (20 mM) was added to the reaction solution, which was then incubated at 37°C for 1 hour, and the reaction was stopped by adding 3N HCl. The Glu concentration in the reaction solution was calculated by measuring the absorbance at 555 nm with a UV-visible spectrophotometer (GeneQuant 1300, Biochrom) using the L-glutamic acid measurement kit "Yamasa" NEO (Yamasa Shoyu Co., Ltd.) according to the manual, and calculated from the following formula.

[0119] L-glutamic acid concentration (mg / L) ={(Sample measurement value - Blank) / (STD measurement value - Blank)} x 250

[0120] 4.Statistical analysis The measured data were expressed as the mean ± standard error. Statistical analysis of the experiment was performed using JMP Pro version 15.2.0 (SAS Institute Inc, NC, USA) with the Tukey-Kramer HSD test. p < 0.05 was considered statistically significant.

[0121] Experimental Example 1: Evaluation of GLS1 inhibitory activity of compounds A to E Five compounds (compounds A to E: see Figure 1-1) were selected as candidate GLS1 inhibitors from a commercially available compound database by performing a primary screening using a 3D pharmacophore obtained from the complex crystal structure of the compound glutaminase and a known GLS1 inhibitor, filtering by Lipinski's "Rule of Five", a 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.

[0122] The GLS1 inhibitory activity of these compounds was evaluated by measuring the glutamic acid (Glu) production rate using a method using human recombinant GLS1. As a positive control compound, DON (6-Diazo-5-oxo-L-norleucine), a known GLS1 inhibitor shown in the formula below, was used.

[0123] [ka]

[0124] DON was dissolved in a solvent (DMSO) to a final concentration of 0.1 mM. Compounds A to E were dissolved in DMSO to a final concentration of 0.1 mM. As a control, the L-glutamic acid concentration (%) in the reaction solution was measured in the same manner as for a system in which only the solvent (DMSO) was added instead of the compound, and the Glu production rate was measured.

[0125] The results are shown in Figure 1-2. As shown in the figure, the L-glutamic acid concentration (%) of all compounds was 25% or less, which was lower than the result of DON at the same concentration (45.3±13.4%) (A: 17.9±6.2%, B: 14.9±5.0%, C: 21.5±9.2%, D: 21.0±7.3%, E: 12.0±4.3%). In particular, compound E had the strongest GLS1 inhibitory activity, reducing Glu production by 88% compared to the control (DMSO).

[0126] Experimental Example 2: Evaluation of GLS1 inhibitory activity of compounds 1 to 24 As in Experimental Example 1, the GLS1 inhibitory activity of compounds 1 to 24 produced by the above-mentioned method was evaluated. The results for Compound E (L-glutamic acid concentration (%) in the reaction solution) and the measurement results for Compounds 1 to 24 are shown in Figure 2. The same amount of DMSO was used as a control group. The compounds were dissolved in DMSO to a final concentration of 0.1 mM. All results are shown as mean ± SD. 2, it was confirmed that all of compounds 1 to 24 have GLS1 inhibitory activity. Among them, compounds 10 and 23 (particularly compound 23) were confirmed to exhibit GLS1 inhibitory activity equal to or stronger than that of compound E.

[0127] Experimental Example 3: GLS1 inhibitory activity potency (IC 50 value) From the results of Experimental Examples 1 and 2, the strength of GLS1 inhibitory activity at 0.1 mM was in the following order: Compound 23>Compound E>DON. Therefore, the GLS1 inhibitory activity of Compound 23, Compound E, and DON was evaluated in the range of 0.001 to 1 mM in the same manner as in Experimental Example 1, and the IC 50 The results are shown in Figure 3. As shown in FIG. 3, compound 23, compound E, and DON all exhibited a concentration-dependent GLS1 inhibitory effect in the range of 0.001 to 1 mM. 50 The values ​​were DON: 182.0 μM, compound E: 86.6 μM, and compound 23: 46.7 μM, respectively.

[0128] Thus, it was confirmed that compound 23 and compound E exhibit stronger GLS1 inhibitory activity than DON, which is known as a GLS1 inhibitor. [Sequence List Free Text]

[0129] SEQ ID NO: 1 is the nucleotide sequence of primer G1U1 (NdeI site) used for the preparation of human recombinant GLS1, SEQ ID NO: 2 is the nucleotide sequence of primer G1-204L1 (BamHI site) used for the preparation of the same, and SEQ ID NO: 3 is the amino acid sequence of the His tag attached to the N-terminus of human recombinant GLS1 prepared in the experimental example.

Claims

1. A glutaminase 1 inhibitor comprising a compound represented by general formula (I), a pharma- ceutically acceptable salt thereof, or a solvate thereof as an active ingredient: 【Chemistry 1】 [In formula (I), X is an optionally substituted 5-membered heterocyclic group having at least two nitrogen atoms; Y is -NH-, -C(=O)-, -(CH ２ ) ｍ A divalent group formed by combining two or three groups selected from the group consisting of -, -O-, and -O-; R １ and R ２ are the same or different and are an alkoxy group or a hydrogen atom; or R １ and R ２ are linked together to form a dioxyalkylene group, and are bonded to adjacent carbon atoms on the phenyl ring to form a 5- or 6-membered fused ring; R ３ represents an optionally substituted phenyl group, an optionally substituted cycloalkyl group, or an alkyl group; n is 0 or 1, and m is 1, 2 or 3.

2. The glutaminase 1 inhibitor according to claim 1 , wherein in the compound of formula (I), X is a thiadiazole residue or a triazole residue.

3. In the compound represented by the formula (I), Y is -NH-C(=O)-, -NH-C(=O)-CH ２ -, -C(=O)-NH-CH ２ - and -CH ２ The glutaminase 1 inhibitor according to claim 1 or 2, wherein the aryl group is a group selected from the group consisting of -O-.

4. In the compound represented by formula (I), R １ and R ２ The glutaminase 1 inhibitor according to claim 1 or 2, wherein one of is an alkoxy group and the other is a hydrogen atom.

5. In the compound represented by formula (I), R ３ The glutaminase 1 inhibitor according to claim 1 or 2, wherein is a phenyl group which may have a substituent.

6. The glutaminase 1 inhibitor according to claim 1 or 2, wherein in the compound represented by formula (I), n is 1.

7. A pharmaceutical composition, a food or beverage composition, or a cosmetic composition, comprising as an active ingredient the glutaminase 1 inhibitor according to any one of claims 1 to 6.

8. The pharmaceutical composition, food or beverage composition, or cosmetic composition according to claim 7, which is used for the prevention, amelioration, or treatment of inflammation, diseases resulting from inflammation, obesity, metabolic syndrome, aging, or cancer.

9. A compound having glutaminase 1 inhibitory activity represented by general formula (II), a pharma- ceutically acceptable salt thereof, or a solvate thereof: 【Chemistry 2】 [In formula (II), R １ and R ２ are the same or different and each is an alkoxy group or a hydrogen atom; R ３ represents a phenyl group, a cycloalkyl group, or an alkyl group, each of which may have a substituent.

10. The R １ and R ２ The compound according to claim 9 , or a pharma- ceutically acceptable salt or solvate thereof, wherein is a hydrogen atom.

Images