Compounds targeting egr-1 and therapeutic uses for atopic dermatitis

Abstract

This invention relates to compounds targeting EGR-1 and their use in treating atopic dermatitis. The compounds targeting EGR-1, their isomers, or pharmaceutically acceptable salts thereof, and compositions comprising them according to the invention reduce EGR-1-mediated inflammatory cytokines by inhibiting the DNA-binding ability of the transcription factor EGR-1, and therefore can be used to prevent, improve, or treat atopic dermatitis.

Description

Technical Field

[0001] This invention relates to compounds targeting EGR-1, isomers thereof, or pharmaceutically acceptable salts thereof, and compositions comprising said compounds for the prevention, improvement, or treatment of atopic dermatitis. Background Technology

[0002] Atopic dermatitis is a chronic, relapsing inflammatory skin disease caused by an allergic immune response, resulting in impaired skin barrier function, dry skin, and severe itching. It has been reported that excessive expression of inflammatory cytokines at the lesion site exacerbates inflammation and itching. Cytokines are proteins secreted by microbial infections or immune-related cells that influence interactions between cells related to the immune system. When cytokine production and secretion are excessive or improperly regulated, they may mediate or exacerbate various diseases related to immune regulatory dysfunction and physiological states such as inflammation.

[0003] In the lesions of acute atopic dermatitis, the levels of thymic stromal lymphopoietin (TSLP) secreted by keratinocytes and interleukin-4 (IL-4), interleukin-13 (IL-13), and interleukin-31 (IL-31) secreted by CD4++ helper type 2 T lymphocytes (Th2) are significantly increased. In contrast, in the lesions of chronic atopic dermatitis, the levels of interferon-gamma (IFNγ) and tumor necrosis factor-alpha (TNFα) secreted by Th1 lymphocytes are increased.

[0004] In the past, the treatment of atopic dermatitis mainly involved topical corticosteroids. Corticosteroids are synthetic adrenocortical hormones that suppress allergic immune responses, reduce cytokine production, and relieve skin itching. However, long-term use of corticosteroids in infants and young children can lead to side effects such as weakened immunity, skin atrophy, or growth retardation. Therefore, non-corticosteroid immunosuppressants, such as the calcineurin inhibitor cyclosporine, have been developed and commercialized. However, long-term use may lead to side effects such as cancer due to weakened biological immunity (J Invest Dermatol 2007; 127:808-816). Therefore, various institutions are conducting research to find substances that can alleviate atopic dermatitis symptoms in ways different from traditional corticosteroids.

[0005] EGR-1 (Early Growth Response Gene-1) is a protein encoded by the EGR-1 gene and is an important transcription factor in cell growth, differentiation, survival, apoptosis, and immune responses. EGR-1 possesses three Cys2-His2 type zinc finger DNA-binding domains. Transcription and gene expression are initiated only after EGR-1 binds to DNA. In other words, EGR-1 regulates the production of proteins, including cytokines.

[0006] EGR-1 regulates inflammatory responses in various tissues, with increased levels in the skin, dermal wound sites, and psoriatic tissues. Furthermore, EGR-1 mediates IL-33-stimulated thymic stromal lymphopoietin (TSLP) production in human keratinocytes and increases the production of the IL-17-induced psoriasin (S100A7), suggesting it may be an important factor in skin inflammation (Exp Dermatol 2015; 24:857-63; Exp Dermatol 2014; 23:890-5).

[0007] Through prior research, the inventors discovered that when skin is exposed to an inflammatory environment, EGR-1, expressed in keratinocytes, is a key transcription factor. It increases the expression of cytokines that induce allergic reactions (such as TSLP, IL-1β, IL-6, IL-17, and IL-23) and chemokines (CXCL1 and CCL5) (attracting T lymphocytes and mast cells to the lesion site), and also increases the expression of the POMC gene. POMC is a precursor to β-endorphin, which stimulates sensory nerves in the skin and transmits itch signals to the brain. Therefore, substances that inhibit the DNA-binding ability of EGR-1 could be an important method for treating atopic dermatitis. Summary of the Invention

[0008] The problem that the invention aims to solve The purpose of this invention is to provide compounds, isomers thereof, or pharmaceutically acceptable salts thereof that target the transcription factor EGR-1.

[0009] Another object of the present invention is to provide a composition for the prevention, improvement or treatment of atopic dermatitis, comprising a compound targeting EGR-1, its isomers or pharmaceutically acceptable salts thereof.

[0010] means for solving problems To achieve the above objectives, the present invention provides a pharmaceutical composition for the prevention, improvement, or treatment of atopic dermatitis, comprising a compound of formula 1, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient: [Formula 1]

[0011] In the above formula, X represents hydrogen, C1-C6 alkyl, or -CH2-C. 6-10 aryl or The C1-C6 alkyl group and -CH2-C 6-10 Each aryl group is independently unsubstituted or substituted by one to three halogens; R 1 It is hydrogen or halogen; R 2 It is hydrogen, halogen, hydroxyl, C1-C6 alkyl or C1-C6 alkoxy, wherein the C1-C6 alkyl and C1-C6 alkoxy are each independently unsubstituted or substituted by 1 to 3 halogens; R 3 R 4 and R 5 Each is independently hydrogen, halogen, hydroxyl or C1-C6 alkoxy, wherein the C1-C6 alkoxy is unsubstituted or substituted by 1 to 3 halogens; R 6 It is hydrogen, halogen, hydroxyl, -NO2, C1-C6 alkyl or C1-C6 alkoxy, wherein the C1-C6 alkyl and C1-C6 alkoxy are each independently unsubstituted or substituted by 1 to 3 halogens; R 7 It is hydrogen, halogen, hydroxyl, -NO2, -CN, C1-C6 alkyl or C1-C6 alkoxy, wherein the C1-C6 alkyl and C1-C6 alkoxy are each independently unsubstituted or substituted by 1 to 3 halogens.

[0012] The present invention provides an over-the-counter pharmaceutical composition comprising a compound of chemical formula 1, an isomer thereof, or a pharmaceutically acceptable salt thereof for the prevention or improvement of atopic dermatitis.

[0013] The present invention also provides a compound, which is a composition selected from novel compounds targeting EGR-1, or isomers thereof or pharmaceutically acceptable salts thereof; (Z)-N-(4-cyanophenyl)-2-(7-fluoro-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(7-fluoro-2-oxoindoline-3-ylidene)-N-(4-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazino-1-thiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(2-methoxyphenyl)hydrazino-1-thiocarbamic acid; (Z)-N-(4-cyanophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(4-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid; (Z)-N-ethyl-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazylthiocarbamate; (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(7-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamate; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(3-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(4-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-chloro-2-oxoindoline-3-ylidene)-N-(2-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(7-fluoro-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(3-(trifluoromethyl)phenyl)hydrazine thiocarbamic acid; (Z)-N-(4-fluorophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid; (Z)-N-(4-bromophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid; (Z)-N-benzyl-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid; (Z)-N-(4-cyanophenyl)-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; and (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-(m-tolyl)hydrazine thiocarbamate.

[0014] The present invention also provides compositions comprising any one of the following compounds, its isomers, or pharmaceutically acceptable salts thereof: (Z)-N-(4-cyanophenyl)-2-(7-fluoro-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(7-fluoro-2-oxoindoline-3-ylidene)-N-(4-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazino-1-thiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(2-methoxyphenyl)hydrazino-1-thiocarbamic acid; (Z)-N-(4-cyanophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(4-(trifluoromethyl)phenyl)thiohydrazine thiocarbamic acid; (Z)-N-ethyl-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazylthiocarbamate; (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazine thiocarbamate; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(7-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(3-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(4-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-chloro-2-oxoindoline-3-ylidene)-N-(2-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(7-fluoro-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(3-(trifluoromethyl)phenyl)hydrazine thiocarbamic acid; (Z)-N-(4-fluorophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid; (Z)-N-(4-bromophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid; (Z)-N-benzyl-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid; (Z)-N-(4-cyanophenyl)-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; and (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-(m-tolyl)hydrazinothiocarbamate.

[0015] Invention Effects The compounds of the present invention targeting EGR-1, their isomers, or pharmaceutically acceptable compositions thereof, comprising such compounds, can be effectively used to prevent, improve, or treat atopic dermatitis by reducing the formation of cytokines that induce skin inflammation through binding to the DNA binding site of the transcription factor EGR-1. Attached Figure Description

[0016] Figure 1 Briefly describe the experimental method of EDBA (EGR-1-DNA binding activity assay).

[0017] Figure 2 This is a graph showing the results of a determination of the DNA binding inhibition rate of a compound against EGR-1 according to an embodiment of the present invention.

[0018] Figure 3 This is a graph showing the results of an in vitro cytotoxicity test of a compound according to an embodiment of the present invention.

[0019] Figure 4 This shows the interaction between compound 21 and the amino acid residues of EGR-1.

[0020] Figure 5 This is a PyMOL-predicted image of the three-dimensional structure of the complex of compound 21 and EGR-1.

[0021] Figure 6 This shows the interaction between compound 23 and the amino acid residues of EGR-1.

[0022] Figure 7 The 3D structure of the complex of compound 23 and EGR-1 is shown using the PyMOL program.

[0023] Figure 8 This shows the interaction between compound 25 and the amino acid residues of EGR-1.

[0024] Figure 9 This is the 3D structure of the complex of compound 25 and EGR-1, obtained using the PyMOL program.

[0025] Figure 10 The results of the molecular docking experiment for compound 21 are shown.

[0026] Figure 11 The results of the molecular docking experiment for compound 23 are shown.

[0027] Figure 12 The results of the molecular docking experiment for compound 25 are shown.

[0028] Figure 13 The results of electrophoretic mobility analysis (EMSA) after applying the compounds of the present invention to HaCaT keratinocytes are shown.

[0029] Figure 14 The results of reverse transcription polymerase chain reaction (RT-PCR) performed after applying a compound of one embodiment of the present invention to HaCaT keratinocytes are shown.

[0030] Figures 15 to 17 The therapeutic effects of compounds 21, 23, and 25 were demonstrated in an animal model of atopic dermatitis. Detailed Implementation

[0031] The present invention will now be described in more detail.

[0032] In this invention, the term "halogen" refers to F, Cl, Br, or I unless otherwise indicated.

[0033] The term "hydroxyl group" refers to the -OH group.

[0034] The term "cyano" refers to the -C≡N group.

[0035] The term "alkyl" refers to a straight-chain or branched saturated hydrocarbon group. For example, "C1-C6 alkyl" has 1 to 6 carbon atoms. Specifically, C1-C6 alkyl includes, but is not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, 1-methylbutyl, n-hexyl, 1-methylpentyl, 2-methylpentyl, 4-methyl-2-pentyl, 3,3-dimethylbutyl, 2-ethylbutyl, etc. In one embodiment, the alkyl group may be independently substituted by one or more substituents, such as 1 to 3 halogens, hydroxyl groups, C1-C6 alkyl groups, and other hydrocarbon groups.

[0036] The term "alkoxy" refers to a group of the formula -O-alkyl, wherein the alkoxy group may be independently substituted by one or more substituents. For example, C1-C6 alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, n-pentoxy, isopentoxy, tert-pentoxy, sec-pentoxy, neopentoxy, hexoxy, etc. In one embodiment, the alkoxy group may be independently substituted by one or more substituents, such as 1 to 3 halogens, hydroxyl groups, C1-C6 alkyl groups, and other hydrocarbon groups.

[0037] The term "aryl" refers to an aromatic ring of a hydrocarbon, whether monocyclic or bicyclic. That is, unless otherwise defined, aryl as used herein may include phenyl, naphthyl, and biaryl groups. In one embodiment of the invention, C6-C... 10 Aryl refers to an aromatic ring having 6 to 10 carbon atoms. In one embodiment, 0, 1, 2, 3, 4, 5, or 6 atoms of each ring of the aryl group may be substituted with substituents.

[0038] The term "substitution" refers to replacing a hydrogen atom in a molecule with a substituent, making the resulting compound chemically stable as long as the valence of the specified atom does not exceed that of the substituent. For example, "group A is substituted by B" can mean that the hydrogen atom bonded to carbon atoms or other atoms that make up the skeleton of group A is replaced by the substituent B, thereby forming a covalent bond between group A and B.

[0039] The term "substituent" refers to another group that is attached to the parent group, and there can be more than one substituent. In the case of multiple substituents, they can be the same or different. When both the parent group and the substituent are hydrocarbon groups, the number of carbon atoms in the parent group does not include the number of carbon atoms in the substituent. For example, butyl (-C4H9) with a methoxy (-O-CH3) substituent is classified as C1 alkoxy, C4 alkyl.

[0040] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. In this document, the singular forms “a,” “an,” and “the” include plural references unless the context otherwise requires. Similar or equivalent methods and materials to those described herein may be used in carrying out or testing the invention, but suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated herein for reference. References cited herein should not be considered precedents to the claimed invention. In the event of any contradiction, this specification, including its definitions, shall prevail. Furthermore, materials, methods, and examples are for illustrative purposes only and not for limitation.

[0041] The present invention provides a pharmaceutical composition for the prevention, improvement or treatment of atopic dermatitis, comprising a compound of formula 1, an isomer or a pharmaceutically acceptable salt as an active ingredient.

[0042] [Formula 1]

[0043] In the formula, X represents hydrogen, C1-C6 alkyl, or -CH2-C. 6-10 aryl or The C1-C6 alkyl and -CH2-C 6-10 The aryl group may be unsubstituted or substituted by 1 to 3 halogens; R 1 It is hydrogen or halogen; R 2 It is hydrogen, halogen, hydroxyl, C1-C6 alkyl or C1-C6 alkoxy, wherein the C1-C6 alkyl and C1-C6 alkoxy are each independently unsubstituted or substituted by 1 to 3 halogens; R 3 R 4 and R 5 Each of the following is independently hydrogen, halogen, hydroxyl, or C1-C6 alkoxy group, wherein the C1-C6 alkoxy group is unsubstituted or substituted by one to three halogens. R 6It is hydrogen, halogen, hydroxyl, -NO2, C1-C6 alkyl or C1-C6 alkoxy, wherein the C1-C6 alkyl and C1-C6 alkoxy are each independently unsubstituted or substituted by 1 to 3 halogens. R 7 It is hydrogen, halogen, hydroxyl, -NO2, -CN, C1-C6 alkyl or C1-C6 alkoxy, wherein the C1-C6 alkyl and C1-C6 alkoxy are each independently unsubstituted or can be substituted by 1 to 3 halogens.

[0044] The present invention also provides an over-the-counter pharmaceutical composition comprising a compound of formula 1, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient for the prevention or improvement of atopic dermatitis.

[0045] [Formula 1]

[0046] In the formula, X represents hydrogen, C1-C6 alkyl, or -CH2-C. 6-10 aryl or The C1-C6 alkyl groups and -CH2-C 6-10 The aryl group may be unsubstituted or substituted by 1 to 3 halogens; R 1 It is hydrogen or halogen; R 2 It is hydrogen, halogen, hydroxyl, C1-C6 alkyl or C1-C6 alkoxy, wherein the C1-C6 alkyl and C1-C6 alkoxy are each independently unsubstituted or substituted by 1 to 3 halogens; R 3 R 4 and R 5 Each is independently hydrogen, halogen, hydroxyl or C1-C6 alkoxy, wherein the C1-C6 alkoxy is unsubstituted or substituted by 1 to 3 halogens; R 6 It is hydrogen, halogen, hydroxyl, -NO2, C1-C6 alkyl or C1-C6 alkoxy, wherein the C1-C6 alkyl and C1-C6 alkoxy are each independently unsubstituted or substituted by 1 to 3 halogens; R 7 It is hydrogen, halogen, hydroxyl, -NO2, -CN, C1-C6 alkyl or C1-C6 alkoxy, wherein the C1-C6 alkyl and C1-C6 alkoxy are each independently unsubstituted or substituted by 1 to 3 halogens.

[0047] In one embodiment, in the compound of Formula 1, X is hydrogen, methyl, ethyl, benzyl, or... The methyl and benzyl groups therein may be independently unsubstituted or substituted by one to three halogens.

[0048] Furthermore, in one embodiment, in the compound of formula 1, R 1 It can be hydrogen or halogen. Specifically, R 1 It can be H, Br, or Cl, but is not limited to these.

[0049] In one embodiment, R in the compound of formula 1 2 It is hydrogen, halogen, hydroxyl, methyl, or methoxy, wherein the methyl and methoxy groups are each independently unsubstituted or substituted by one to three halogens. For example, R 2 It can be H, F, Br, CH3, -OCH3, or -OCF3, but is not limited to these.

[0050] In one embodiment, R in the compound of formula 1 3 It is hydrogen, halogen, hydroxyl, or methoxy, wherein the methoxy group may be independently unsubstituted or substituted by one to three halogens. For example, R 3 It can be H, Br, Cl or -OCH3, but is not limited to these.

[0051] In one embodiment, R in the compound of formula 1 4 It can be hydrogen, halogen, or hydroxyl. For example, R 4 It can be H, F, or Br, but is not limited to these.

[0052] In one embodiment, R in the compound of formula 1 5 It is hydrogen, hydroxyl, or methoxy, wherein the methoxy group may be independently unsubstituted or substituted by one to three halogens. For example, R 5 It can be H or -OCH3, but is not limited to these.

[0053] In one embodiment, R in the compound of formula 1 6 It is hydrogen, halogen, hydroxyl, -NO2, methyl, or methoxy, wherein the methyl and methoxy groups may each be unsubstituted or substituted with one to three halogens. For example, R 6 It can be H, -NO2, CH3, -OCH3, or CF3, but is not limited to these.

[0054] In one embodiment, R in the compound of formula 1 7 It is hydrogen, halogen, hydroxyl, -NO2, -CN, methyl, or methoxy, wherein the methyl and methoxy groups are either unsubstituted or substituted by one to three halogens. For example, R 7 It can be H, F, Br, -NO2, -CN, -OCH3, or CF3, but is not limited to these.

[0055] Furthermore, specific examples of compounds of Formula 1 are as follows, but not limited thereto.

[0056] N-(3-(trifluoromethyl)phenyl)hydrazinothiocarbamate; N-(4-fluorophenyl)hydrazinothiocarbamate; N-(4-bromophenyl)hydrazinothiocarbamate; N-(4-cyanophenyl)hydrazinothiocarbamate; N-(4-trifluoromethylphenyl)hydrazinothiocarbamate; N-Ethylhydrazinothiocarbamate; N-Isopropylhydrazine thiocarbamate; N-Isobutylhydrazine thiocarbamate; N-(hydrazylaminothioformyl)benzamide; N-Benzylhydrazine thiocarbamate; N-(m-tolyl)hydrazinothiocarbamate; (Z)-2-(5-fluoro-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(5-fluoro-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(5-fluoro-2-oxoindoline-3-ylidene)-N-(4-fluorophenyl)hydrazylthiocarbamic acid; (Z)-N-(4-bromophenyl)-2-(5-fluoro-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-N-(4-cyanophenyl)-2-(5-fluoro-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(5-fluoro-2-oxoindoline-3-ylidene)-N-(4-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid; (Z)-2-(7-fluoro-2-oxoindoline-3-ylidene)-N-(4-fluorophenyl)hydrazylthiocarbamic acid; (Z)-N-(4-bromophenyl)-2-(7-fluoro-2-oxoindoline-3-ylidene)hydrazylthiocarbamate; (Z)-N-(4-cyanophenyl)-2-(7-fluoro-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(7-fluoro-2-oxoindoline-3-ylidene)-N-(4-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazino-1-thiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(2-methoxyphenyl)hydrazino-1-thiocarbamic acid; (Z)-N-(4-cyanophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(4-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid; (Z)-N-ethyl-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazylthiocarbamate; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(m-tolyl)hydrazinothiocarbamic acid; (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-N-(2-methoxyphenyl)-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-(3-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid; (Z)-N-(4-fluorophenyl)-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(5-bromo-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(5-bromo-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-N-(2-methoxyphenyl)-2-(2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(7-bromo-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-hydrazylthiocarbamic acid; (Z)-2-(5-bromo-2-oxoindoline-3-ylidene)-N-hydrazylthiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-hydrazylthiocarbamic acid; (Z)-2-(7-bromo-2-oxoindoline-3-ylidene)-N-hydrazylthiocarbamic acid; (Z)-N-(3-nitrophenyl)-2-(2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(5-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(7-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-N-(3-methoxyphenyl)-2-(2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(3-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(5-bromo-2-oxoindoline-3-ylidene)-N-(3-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-N-(4-methoxyphenyl)-2-(2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(4-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(5-bromo-2-oxoindoline-3-ylidene)-N-(4-methoxyphenyl)hydrazylthiocarbamic acid; N-(3,4-dimethoxyphenyl)hydrazinothiocarbamate; (Z)-N-(4-fluorophenyl)-2-(2-oxoindoline-3-ylidene)hydrazylthiocarbamate; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-chloro-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(6-chloro-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(4-chloro-2-oxoindoline-3-ylidene)-N-(2-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(5-fluoro-2-oxoindoline-3-ylidene)-N-(3-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid; (Z)-N-benzyl-2-(5-fluoro-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(5-fluoro-2-oxoindoline-3-ylidene)-N-(m-tolyl)hydrazinothiocarbamic acid; (Z)-2-(7-fluoro-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(3-(trifluoromethyl)phenyl)hydrazine thiocarbamic acid; (Z)-N-(4-fluorophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid; (Z)-N-(4-bromophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid; (Z)-N-benzyl-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid; (Z)-N-(4-bromophenyl)-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-N-(4-cyanophenyl)-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-N-ethyl-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamate; (Z)-N-benzyl-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-(m-tolyl)hydrazinothiocarbamic acid; (Z)-2-(2-oxo-5-(trifluoromethoxy)indoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-N-(3-nitrophenyl)-2-(2-oxo-5-(trifluoromethoxy)indoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-N-(4-fluorophenyl)-2-(2-oxo-5-(trifluoromethoxy)indoline-3-ylidene)hydrazylthiocarbamate; and (Z)-N-(4-bromophenyl)-2-(2-oxo-5-(trifluoromethoxy)indoline-3-ylidene)hydrazylthiocarbamate.

[0057] This invention includes pharmaceutically acceptable salts of the above-mentioned compounds.

[0058] The pharmaceutically acceptable salt should be non-toxic to humans and should not negatively affect the biological activity and physicochemical properties of the parent compound. For example, the pharmaceutically acceptable salt can be an acid addition salt formed from a pharmaceutically acceptable free acid.

[0059] As a free acid, either inorganic or organic acids can be used. In this case, inorganic acids can be hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, perchloric acid, and bromic acid, while organic acids can be acetic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, fumaric acid, maleic acid, malonic acid, phthalic acid, succinic acid, lactic acid, citric acid, gluconic acid, tartaric acid, salicylic acid, malic acid, oxalic acid, benzoic acid, ethyl acetate, aspartic acid, and glutamic acid, etc.

[0060] Acid addition salts can be prepared by conventional methods, such as by dissolving the compound of formula 1 in an excess aqueous solution and then precipitating the salt with a miscible organic solvent such as methanol, ethanol, acetone or acetonitrile.

[0061] The pharmaceutically acceptable salt may be an alkali metal salt (e.g., sodium salt) or an alkaline earth metal salt (e.g., potassium salt).

[0062] Alkali metal salts or alkaline earth metal salts can be prepared, for example, by dissolving the compound of Formula 1 in an excess of alkali metal hydroxide solution or alkaline earth metal hydroxide solution, filtering out the undissolved compound, and then evaporating and drying the filtrate.

[0063] The compounds of the present invention may also have a chiral carbon atom center and may exist in the form of R or S isomers, racemates, individual enantiomers or mixtures thereof, individual diastereomers or mixtures thereof, and all such stereoisomers and mixtures thereof are within the scope of the present invention.

[0064] The compounds of the present invention may also include hydrates and solvates of the compounds of Formula 1 above. The hydrates and solvates can be prepared using known methods, and are preferably non-toxic and water-soluble. Specifically, the hydrates and solvates can be combined with 1 to 5 molecules of water or an alcohol solvent (especially ethanol, etc.), respectively.

[0065] The compounds, isomers thereof, or pharmaceutically acceptable salts thereof shown in the embodiments of the present invention have excellent effects on inhibiting EGR-1 activity, and therefore, compounds thereof or pharmaceutical compositions thereof can be used to treat atopic dermatitis caused by an increase in EGR-1-mediated inflammatory cytokines.

[0066] Specifically, the compounds of the present invention, or pharmaceutical compositions comprising them, are substances that target the transcription factor EGR-1, which is involved in the immune system, for example, by inhibiting the DNA-binding ability of EGR-1 to reduce the production and secretion of inflammatory cytokines. Therefore, skin inflammation and itching caused by excessive immune responses can be improved.

[0067] By replacing existing steroid preparations with the above-mentioned compounds, it is possible to prevent the decrease in immunity and other side effects caused by conventional steroid series of therapeutic agents, and they can be used as a treatment for atopic dermatitis.

[0068] As used in this article, the term "atopic dermatitis" can be used interchangeably with "atopic," "atopic disease," etc., and can refer to a chronic allergic inflammatory disease of the skin, accompanied by symptoms such as dry, scaly skin, skin inflammation, increased skin permeability, susceptibility to skin surface infections, flare-ups, and itching.

[0069] As used herein, the term “prevention” means any action that inhibits or delays the occurrence, spread, and recurrence of a disease by administering the compounds or pharmaceutical compositions of the present invention; the term “improvement” means any action that at least reduces parameters (e.g., the severity of symptoms) associated with the relief or cure of a disease state by administering the compounds or pharmaceutical compositions of the present invention; and the term “treatment” means any action that improves or beneficially alters the symptoms of the disease by administering the compounds or pharmaceutical compositions of the present invention, including inhibiting, reducing, or eliminating the occurrence of the disease.

[0070] This invention provides the use of EGR-1 targeting compounds, their isomers, or pharmaceutically acceptable salts thereof in the prevention, improvement, or treatment of atopic dermatitis.

[0071] This invention provides the use of compounds targeting EGR-1, their isomers, or pharmaceutically acceptable salts thereof to inhibit the activity of the transcription factor EGR-1.

[0072] This invention provides the use of compounds targeting EGR-1, their isomers, or pharmaceutically acceptable salts thereof in the preparation of medicaments for the prevention, improvement, or treatment of atopic dermatitis.

[0073] This invention provides a method for preparing an EGR-1-inhibiting drug comprising a compound targeting EGR-1, its isomers, or pharmaceutically acceptable salts thereof.

[0074] The present invention also provides methods for preventing, improving or treating atopic dermatitis, including administering a compound, an isomer thereof or a pharmaceutically acceptable salt thereof to a subject requiring the compound to target EGR-1.

[0075] The present invention also provides a method for inhibiting EGR-1 activity, comprising administering to a subject requiring the treatment a compound, an isomer thereof, or a pharmaceutically acceptable salt thereof that targets EGR-1.

[0076] The pharmaceutical compositions of the present invention can inhibit the activity of EGR-1 and the resulting production of cytokines. As used herein, the term "inhibition" means the inhibition of any step of gene transcription, mRNA processing, translation, translocation, and maturation, or the inhibition of protein-protein binding, protein activation, or signal transduction.

[0077] In addition to the active ingredient, the pharmaceutical composition of the present invention may also include a pharmaceutically acceptable carrier. In this case, the pharmaceutically acceptable carrier is a carrier commonly used in formulation and may include, but is not limited to, lactose, dextran, sucrose, sorbitol, glycerin, starch, gum arabic, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylparaben, propylparaben, talc, magnesium stearate, and mineral oil. Furthermore, in addition to the above-mentioned components, it may further include lubricants, humectants, sweeteners, flavorings, emulsifiers, suspending agents, preservatives, etc.

[0078] The pharmaceutical compositions of the present invention can be administered orally or via non-enteral methods according to desired procedures. Non-enteral administration includes intravenous, subcutaneous, intraperitoneal, intramuscular injection, and topical application (e.g., skin application). The dosage depends on the patient's condition and weight, disease severity, drug form, route of administration, and timing, and can be appropriately selected by those skilled in the art.

[0079] The pharmaceutical compositions of the present invention are administered in pharmaceutically effective amounts. In the present invention, a "pharmaceutically effective amount" means an amount sufficient to treat a disease at an acceptable risk / benefit ratio. The effective dose level can be determined based on factors including the patient's disease type, severity, drug activity, sensitivity to the drug, timing of administration, route of administration and clearance rate, duration of treatment, and other drugs used concurrently, as well as other factors commonly known in the medical field.

[0080] The pharmaceutical compositions of the present invention can be administered as a single therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered in single or multiple doses. Importantly, considering all factors, the goal is to administer the minimum amount to achieve the maximum effect without side effects, which is readily determined by those skilled in the art.

[0081] The present invention also provides a method for improving, regulating, or treating atopic dermatitis, comprising applying the above-described pharmaceutical composition to an individual. In this specification, "individual" refers to a person who requires treatment for a disease, and more specifically, to humans or non-human primates, such as mammals like rats, dogs, cats, horses, and cattle.

[0082] In one embodiment, the pharmaceutical composition of the present invention can be a topical skin formulation. The topical skin formulation can be a cream, gel, ointment, skin emulsifier, skin suspension, transdermal patch formulation, lotion, or a combination thereof. The topical skin formulation can be suitably used in combination with ingredients commonly used in cosmetics or pharmaceuticals and other topical skin formulations (e.g., aqueous ingredients, oily ingredients, powder ingredients, alcohols, moisturizers, thickeners, UV absorbers, whitening agents, preservatives, antioxidants, surfactants, fragrances, pigments, various skin nutrients, or combinations thereof). The topical skin preparation can also be used in combination with metal chelating agents such as sodium EDTA, trisodium EDTA, sodium citrate, sodium polyphosphate, sodium metaphosphate, and sodium gluconate, as well as caffeine, tannic acid, berapamil, licorice extract, glabridin, hot water extract of calin fruit, various crude drugs, tocopherol acetate, glycyrrhizic acid, tranilast and its derivatives or their salts, vitamin C, magnesium ascorbate phosphate, ascorbate glucoside, arbutin, kojic acid, glucose, fructose, trehalose, and other sugars.

[0083] Skin includes all areas of the body, including the face, hands, arms, legs, feet, chest, abdomen, back, buttocks, and scalp.

[0084] In this specification, the term "non-prescription drug" refers to an article that is less severe than a medicine and is intended for the diagnosis, treatment, improvement, relief, management, or prevention of diseases in humans or animals. For example, according to the Korean Pharmacist Act, non-prescription drugs are articles other than those used as medicines, including fiber and rubber products used to treat or prevent diseases in humans or animals, articles that have a slight or indirect effect on the human body and are not instruments or machines, as well as bactericides and insecticides used to prevent infectious diseases.

[0085] The types and dosage forms of the over-the-counter pharmaceutical compositions of the present invention are not particularly limited, and may include bandages, gauze, absorbent cotton, adhesive bandages, disinfectants, bath foam, mouthwash, wet wipes, detergents, hand sanitizers, humidifier fillers, masks, or filter fillers.

[0086] The over-the-counter compositions according to the present invention may have the effect of preventing or improving atopic dermatitis, but are not limited thereto.

[0087] In addition to the above-mentioned components, the pharmaceutical composition of the present invention may also contain, as needed, a pharmaceutically acceptable carrier, excipient, or diluent. There are no limitations on the pharmaceutically acceptable carrier, excipient, or diluent, provided that it does not impair the effects of the present invention; for example, it may include fillers, extenders, binders, wetting agents, disintegrants, surfactants, lubricants, sweeteners, flavorings, preservatives, etc.

[0088] When used as an over-the-counter medicine for the prevention or improvement of allergic dermatitis, the compositions of the present invention can be used alone or in combination with other over-the-counter medicine ingredients. Any conventional method can be used. The mixing amount of the active ingredients can be appropriately determined according to the intended use.

[0089] This invention provides a method for preparing the above-described compounds. Specifically, the compounds of Formula 1 can be prepared by the method shown in the following scheme, but are not limited thereto. In particular, those skilled in the art will fully understand that the compounds of Formula 1 can be prepared in various ways using techniques known in the art.

[0090] The following scheme illustrates a method for preparing representative compounds of the present invention, as an example of preparation steps. Many compounds of the present invention can be prepared by changing the reagents and solvents used in the following preparation steps or by changing the reaction order.

[0091] General Procedure In this invention, the compounds of Formula 1 can be prepared by the method shown in Scheme 1. Hydrazine thiocarbamates (A and B) are obtained by reacting various isothiocyanates having chain or ring substituents with hydrazine monohydrate (step 1), and then hydrazine thioamides (A and B) are reacted with isoindole (C) to obtain compounds of Formula D (step 2).

[0092] Option (1)

[0093] The desired compounds prepared in Scheme 1 can be separated and purified using conventional methods, such as column chromatography, recrystallization, etc.

[0094] On the other hand, the present invention provides a novel compound for inhibiting EGR-1 activity, the compound being selected from any one of the following groups, its isomers, or pharmaceutically acceptable salts thereof: (Z)-N-(4-cyanophenyl)-2-(7-fluoro-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(7-fluoro-2-oxoindoline-3-ylidene)-N-(4-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazino-1-thiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(2-methoxyphenyl)hydrazino-1-thiocarbamic acid; (Z)-N-(4-cyanophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(4-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid; (Z)-N-ethyl-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazylthiocarbamate; (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(7-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(3-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(4-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-chloro-2-oxoindoline-3-ylidene)-N-(2-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(7-fluoro-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(3-(trifluoromethyl)phenyl)hydrazine thiocarbamic acid; (Z)-N-(4-fluorophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid; (Z)-N-(4-bromophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid; (Z)-N-benzyl-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazylthio-1-carbamic acid; (Z)-N-(4-cyanophenyl)-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; and (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-(m-tolyl)hydrazinothiocarbamate.

[0095] The novel compounds, isomers thereof, or pharmaceutically acceptable salts thereof that inhibit EGR-1 activity as described in this invention are as described above.

[0096] The present invention also provides compositions comprising any one of the following compounds, its isomers, or pharmaceutically acceptable salts thereof: (Z)-N-(4-cyanophenyl)-2-(7-fluoro-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(7-fluoro-2-oxoindoline-3-ylidene)-N-(4-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazino-1-thiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(2-methoxyphenyl)hydrazino-1-thiocarbamic acid; (Z)-N-(4-cyanophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(4-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid; (Z)-N-ethyl-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazylthiocarbamate; (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(7-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(3-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(4-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-chloro-2-oxoindoline-3-ylidene)-N-(2-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(7-fluoro-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(3-(trifluoromethyl)phenyl)hydrazine thiocarbamic acid; (Z)-N-(4-fluorophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid; (Z)-N-(4-bromophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid; (Z)-N-benzyl-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid; (Z)-N-(4-cyanophenyl)-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; and (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-(m-tolyl)hydrazinothiocarbamate.

[0097] The present invention will now be described in more detail through examples and experimental cases. However, the examples and experimental cases described below are merely illustrative of the present invention and do not limit the scope of the present invention.

[0098] Therefore, the method for preparing compound 1 is illustrated using the synthetic process of N-benzylhydrazine thiourea (compound 10), one of the class VI compounds, and (Z)-N-(4-cyanophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid (compound 25), one of the class VII compounds.

[0099] <Example> Example 1. Synthesis of N-benzylhydrazine thiocarbamic acid (compound 10)

[0100] Benzyl isothiocyanate (1.49 g, 10 mmol) was added to a round-bottom flask and dissolved in 40 mL of ethanol. Hydrazine monohydrate (0.6 g, 12 mmol) was added to the solution at room temperature, and the reaction mixture was heated and stirred at 90 °C for 8 hours. The reaction was confirmed to be complete by thin-layer chromatography. The reaction mixture was cooled, and the solvent was removed by distillation under reduced pressure to give a solid product. Recrystallization of this compound gave the title compound (1.37 g, 76%).

[0101] 1H NMR (700 MHz, dmso) δ 8.74 (s, 1H), 8.30 (s, 1H), 7.35 - 7.26 (m, 4H), 7.25 - 7.17 (m, 1H), 4.71 (d, J = 6.0 Hz, 2H), 4.51 (s, 2H). 13C NMR (175 MHz, dmso) δ 181.57, 139.78, 128.05, 127.30, 126.61, 46.16. ESI-MS. Calculated value for C8H11N3S (M+ +H): m / z 182.0674. Measured value: m / z 182.0767.

[0102] Example 2. Synthesis of (Z)-N-(4-cyanophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid (compound 25)

[0103] The method of Example 1 was used to react 4-cyanophenyl isothiocyanate with hydrazine hydrate to give the synthetic intermediate N-(4-cyanophenyl)hydrazinothiocarbamate. N-(4-cyanophenyl)hydrazinothiocarbamate (135 mg, 0.7 mmol) was placed in a round-bottom flask and dissolved in 15 mL of ethanol. 6-methoxyisoquinoline ketone (89 mg, 0.5 mmol) was added to the above solution and dissolved. Then, 2 mL of 3N hydrochloric acid solution was added, and the mixture was stirred at room temperature for 5 hours. As the reaction proceeded, an orange solid formed. After the reaction was completed, the mixture was filtered under reduced pressure to give the title compound (138 mg, 79%).

[0104] 1H NMR (700 MHz, dmso) δ 12.83 (s, 1H), 11.26 (s, 1H), 10.90 (s, 1H), 7.97 (t, J = 9.6 Hz, 2H), 7.93 - 7.82 (m, 2H), 7.68 (d, J = 8.4 Hz, 1H), 6.79- 6.63 (m, 1H), 6.47 (dd, J = 32.7, 2.2 Hz, 1H), 3.82 (d, J = 10.5 Hz, 3H).13C NMR (175 MHz, dmso) δ 175.80, 163.50, 162.86, 144.72, 142.99, 133.14, 132.64, 125.09, 123.29, 118.89, 112.15, 108.42, 107.62, 97.70, 55.76. ESI-MS for C17H13N5O2S (M+ +H): m / z 352.0790. The measured m / z is 352.0877.

[0105] Example 3. Synthesis of N-(3-(trifluoromethyl)phenyl)hydrazinothiocarbamic acid (compound 21) The title compound was synthesized using 3-(trifluoromethyl)phenyl isothiocyanate and hydrazine as starting materials, according to the synthesis method of Example 1.

[0106] 1H NMR (700 MHz, dmso) δ 9.36 (s, 1H), 8.25 (s, 1H), 7.87 (m, 2H), 7.51 (m, 1H), 7.42 (m, 1H), 4.90 (s, 2H). Example 4. Synthesis of N-(4-fluorophenyl)hydrazylthiocarbamic acid (compound 23) The title compound was synthesized using 4-fluorophenyl isothiocyanate and hydrazine as starting materials, according to the synthesis method of Example 1.

[0107] 1H NMR (700 MHz, dmso) δ 9.69 (s, 1H), 9.14 (s, 1H), 7.60 (s, 2H), 7.12 (m, 2H), 4.76 (s, 2H). Example 5. Synthesis of N-(4-bromophenyl)hydrazinothiocarbamic acid (compound 25) The title compound was synthesized using 4-bromophenyl isothiocyanate and hydrazine as starting materials, according to the synthesis method in Example 1.

[0108] 1H NMR (700 MHz, dmso) δ 9.70 (s, 1H), 9.24 (s, 1H), 7.65 (d, J = 7.5Hz, 2H), 7.46 (d, J = 7.5 Hz, 2H), 4.80 (s, 2H). Example 6. Synthesis of N-(4-cyanophenyl)hydrazinothiocarbamic acid (compound 4) The title compound was synthesized using 4-cyanophenyl isothiocyanate and hydrazine as starting materials, according to the synthesis method in Example 1.

[0109] 1H NMR (700 MHz, dmso) δ 9.50 (s, 1H), 8.05 (d, J = 7.9 Hz, 2H), 7.78 (d, J = 7.9 Hz, 2H), 7.64 (s, 1H), 5.09 (s, 2H). Example 7. Synthesis of N-(4-(trifluoromethyl)phenyl)hydrazinothiocarbamic acid (compound 5) The title compound was synthesized using 4-(trifluoromethyl)phenyl isothiocyanate and hydrazine as starting materials, according to the synthesis method of Example 1.

[0110] 1H NMR (700 MHz, dmso) δ 9.40 (s, 1H), 7.98 (s, 2H), 7.70 - 7.58 (m, 3H), 4.93 (s, 2H). Example 8. Synthesis of N-ethylhydrazylthiocarbamic acid (compound 6) The title compound was synthesized using ethyl isothiocyanate and hydrazine as starting materials according to the synthesis method in Example 1.

[0111] 1H NMR (700MHz, dmso) δ 8.49 (s, 1H), 7.81 (s, 1H), 4.42 (s, 2H), 3.45 (q, J = 7.1 Hz, 2H), 1.06 (t, J = 7.1 Hz, 3H). Example 9. Synthesis of N-isopropylhydrazylthiocarbamic acid (compound 7) The title compound was synthesized using isopropyl isothiocyanate and hydrazine as starting materials according to the synthesis method in Example 1.

[0112] 1H NMR (700MHz, dmso) δ 8.52 (s, 1H), 7.48 (s, 1H), 4.42 (s, 2H), 4.35 (m, J = 6.6 Hz, 1H), 1.12 (d, J = 6.6 Hz, 6H). Example 10. Synthesis of N-isobutylhydrazine thiocarbamic acid (compound 8) The title compound was synthesized using isobutyl isothiocyanate and hydrazine as starting materials according to the synthesis method in Example 1.

[0113] 1H NMR (700MHz, dmso) δ 8.56 (s, 1H), 7.80 (s, 1H), 4.46 (s, 2H), 3.27 (m, 2H), 1.92 - 1.80 (m, 1H), 0.85 (d, J = 6.7 Hz, 6H). Example 11. Synthesis of N-(hydrazylaminothioformyl)benzamide (compound 9) The title compound was synthesized using benzoyl isothiocyanate and hydrazine as starting materials according to the synthesis method of Example 1.

[0114] 1H NMR (700MHz, dmso) δ 12.75 (s, 1H), 12.10 - 9.22 (m, 3H), 8.12 (d, J = 7.4 Hz, 2H), 7.67 (t, J = 7.4 Hz, 1H), 7.57 (t, J = 7.4 Hz, 2H). Example 12. Synthesis of N-(m-tolyl)hydrazinothiocarbamic acid (compound 11) The title compound was synthesized using m-toluene isothiocyanate and hydrazine as starting materials, according to the synthesis method of Example 1.

[0115] 1H NMR (700 MHz, dmso) δ 9.62 (s, 1H), 9.10 (s, 1H), 7.45 (s, 2H), 7.21 (m, 1H), 6.92 (m, 1H), 4.77 (s, 2H), 2.28 (s, 3H). Example 13. Synthesis of (Z)-2-(5-fluoro-2-oxoindoline-3-ylidene)-N-phenylhydrazylthiocarbamic acid (compound 12) The title compound was synthesized using phenylhydrazine thiocarbamate and 5-fluoroisosorbide as starting materials, following the synthesis method of Example 2.

[0116] 1H NMR (700 MHz, dmso) δ 12.67 (s, 1H), 11.26 (s, 1H), 10.86 (s, 1H), 7.64 (dd, J = 8.0, 2.6 Hz, 1H), 7.60 (d, J = 7.5 Hz, 2H), 7.44 (t, J = 8.0Hz, 2H), 7.29 (t, J = 7.5 Hz, 1H), 7.21 (m, 1H), 6.94 (dd, J = 8.6, 4.1 Hz, 1H). Example 14. Synthesis of (Z)-2-(5-fluoro-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid (compound 13) The title compound was synthesized from N-(3-nitrophenyl)hydrazinoaminothioformamide and 5-fluoroisosorbone as starting materials, according to the synthesis method in Example 2.

[0117] 1H NMR (700 MHz, dmso) δ 12.83 (s, 1H), 11.31 (s, 1H), 11.10 (s, 1H), 8.65 (t, J = 2.2 Hz, 1H), 8.20 - 8.15 (m, 1H), 8.13 (ddd, J = 8.3, 2.2, 0.9Hz, 1H), 7.73 (t, J = 8.3 Hz, 1H), 7.61 (dd, J = 8.0, 2.5 Hz, 1H), 7.23 (td, J = 9.0, 2.7 Hz, 1H), 6.95 (dd, J = 8.8, 4.1 Hz, 1H). Example 15. Synthesis of (Z)-2-(5-fluoro-2-oxoindoline-3-ylidene)-N-(4-fluorophenyl)hydrazylthiocarbamic acid (compound 14) The title compound was synthesized using N-(4-fluorophenyl)hydrazinothiocarbamic acid and 5-methylisoxazole as starting materials, according to the synthesis method in Example 2.

[0118] 1H NMR (700 MHz, dmso) δ 12.68 (s, 1H), 11.27 (s, 1H), 10.86 (s, 1H), 7.74 - 7.50 (m, 3H), 7.28 (t, J = 9.0 Hz, 2H), 7.21 (td, J = 9.0, 2.7 Hz, 1H), 6.94 (dd, J = 8.6, 4.1 Hz, 1H) Example 16. Synthesis of (Z)-N-(4-bromophenyl)-2-(5-fluoro-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid (compound 21) The title compound was synthesized using N-(4-bromophenyl)hydrazinothiocarbamic acid and 5-methylisoxazole as starting materials, according to the synthesis method in Example 2.

[0119] 1H NMR (700 MHz, dmso) δ 12.72 (s, 1H), 11.28 (s, 1H), 10.87 (s, 1H), 7.70 - 7.54 (m, 5H), 7.22 (td, J = 9.2, 2.7 Hz, 1H), 6.94 (dd, J = 8.6, 4.1Hz, 1H). Example 17. Synthesis of (Z)-N-(4-cyanophenyl)-2-(5-fluoro-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid (compound 23) The title compound was synthesized using N-(4-cyanophenyl)hydrazinothiocarbamic acid and 5-methylisoxazole as starting materials, according to the synthesis method in Example 2.

[0120] 1H NMR (700 MHz, dmso) δ 12.83 (s, 1H), 11.30 (s, 1H), 11.01 (s, 1H), 7.97 (d, J = 8.7 Hz, 2H), 7.91 (d, J = 8.7 Hz, 2H), 7.62 (dd, J = 8.0, 2.6Hz, 1H), 7.23 (td, J = 9.4, 2.7 Hz, 1H), 6.95 (dd, J = 8.6, 4.1 Hz, 1H). Example 18. Synthesis of (Z)-2-(5-fluoro-2-oxoindoline-3-ylidene)-N-(4-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid (compound 25) The title compound was synthesized from N-(4-(trifluoromethyl)phenyl)hydrazinothiocarbamic acid and 5-methylisoxazole as starting materials, according to the synthesis method in Example 2.

[0121] 1H NMR (700 MHz, dmso) δ 12.80 (s, 1H), 11.30 (s, 1H), 11.02 (s, 1H), 7.93 (d, J = 8.4 Hz, 2H), 7.81 (d, J = 8.5 Hz, 2H), 7.63 (dd, J = 8.6, 2.6Hz, 1H), 7.23 (m, 1H), 6.95 (dd, J = 8.6, 4.1 Hz, 1H). Example 19. Synthesis of (Z)-2-(7-fluoro-2-oxoindoline-3-ylidene)-N-(4-fluorophenyl)hydrazylthiocarbamic acid (compound 18) The title compound was synthesized from N-(4-fluorophenyl)hydrazinothiocarbamic acid and 7-fluoroisosorbic acid using the synthesis method of Example 2.

[0122] 1H NMR (700 MHz, dmso) δ 12.77 (s, 1H), 11.80 (s, 1H), 10.90 (s, 1H), 7.64 - 7.56 (m, 3H), 7.33 - 7.24 (m, 3H), 7.13 (m, 1H). Example 20. Synthesis of (Z)-N-(4-bromophenyl)-2-(7-fluoro-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid (compound 19) The title compound was synthesized using N-(4-bromophenyl)hydrazinothiocarbamic acid and 7-fluoroisosorbic acid as starting materials, according to the synthesis method in Example 2.

[0123] 1H NMR (700 MHz, dmso) δ 12.80 (s, 1H), 11.81 (s, 1H), 10.92 (s, 1H), 7.61 (m, 5H), 7.30 (m, 1H), 7.13 (m, 1H). Example 21. Synthesis of (Z)-N-(4-cyanophenyl)-2-(7-fluoro-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid (compound 20) The title compound was synthesized from N-(4-cyanophenyl)hydrazinothiocarbamic acid and 7-fluoroisosorbic acid using the synthesis method of Example 2.

[0124] 1H NMR (700 MHz, dmso) δ 12.91 (s, 1H), 11.83 (s, 1H), 11.07 (s, 1H), 7.98 - 7.94 (m, 2H), 7.92 - 7.88 (m, 2H), 7.62 (m, 1H), 7.32 (m, 1H), 7.14 (m, 1H). Example 22. Synthesis of (Z)-2-(7-fluoro-2-oxoindoline-3-ylidene)-N-(4-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid (compound 21) The title compound was synthesized from N-(4-(trifluoromethyl)phenyl)hydrazinothiocarbamic acid and 7-fluoroisophthalic acid as starting materials, according to the synthesis method in Example 2.

[0125] 1H NMR (700 MHz, dmso) δ 12.88 (s, 1H), 11.83 (s, 1H), 11.07 (s, 1H), 7.92 (d, J = 8.4 Hz, 2H), 7.80 (d, J = 8.5 Hz, 2H), 7.63 (d, J = 7.5 Hz, 1H), 7.31 (m, 1H), 7.14 (m, 1H). 13C NMR (175 MHz, dmso) δ 176.48, 162.64, 147.71, 146.32, 142.21, 132.24 (d, J = 4.9 Hz), 129.51 (d, J = 13.3 Hz), 126.12 (d, J = 32.1 Hz), 125.72, 125.63 (q, J = 3.7 Hz), 125.05, 123.51, 123.42, 122.89 (d, J = 4.7 Hz), 118.33 (d, J = 17.5 Hz), 117.64.

[0126] ESI-MS. For C17H13N5O2S (M+ +H): m / z 383.0511. Measured value: m / z 383.0605.

[0127] Example 23. Synthesis of (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid (compound 22) The title compound was synthesized using phenylhydrazine thiocarbamic acid and 6-methoxyisophthalic acid as starting materials, following the synthesis method of Example 2.

[0128] 1H NMR (700 MHz, dmso) δ 12.67 (s, 1H), 11.22 (s, 1H), 10.70 (s, 1H), 7.68 (d, J = 8.4 Hz, 1H), 7.60 (d, J = 7.8 Hz, 2H), 7.42 (t, J = 7.9 Hz, 2H), 7.26 (m, 1H), 6.69 (dd, J = 8.4, 2.2 Hz, 1H), 6.51 - 6.47 (m, 1H), 3.81 (s, 3H). Example 24. Synthesis of (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazino-1-thiocarbamic acid (compound 23) The title compound was synthesized using N-(3-nitrophenyl)hydrazinothiocarbamic acid and 6-methoxyisophthalic acid as starting materials, according to the synthesis method in Example 2.

[0129] 1H NMR (700 MHz, dmso) δ 12.83 (s, 1H), 11.25 (s, 1H), 10.95 (s, 1H), 8.67 (s, 1H), 8.18 (dd, J = 8.0, 2.0 Hz, 1H), 8.13 - 8.06 (m, 1H), 7.69 (m, 2H), 6.71 (dd, J = 8.4, 2.0 Hz, 1H), 6.47 (d, J = 2.0 Hz, 1H), 3.82 (s, 3H).13C NMR (175 MHz, dmso) δ 176.13, 163.39, 162.74, 147.41, 144.60, 139.75, 133.01, 131.41, 129.55, 123.08, 120.25, 119.43, 112.08, 108.30, 97.60, 55.65.

[0130] ESI-MS for C16H13N5O4S (M+ +H): m / z 372.0688. Measured value: m / z 372.0773.

[0131] Example 25. Synthesis of (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(2-methoxyphenyl)hydrazino-1-thiocarbamic acid (compound 24) The title compound was synthesized using N-(2-methoxyphenyl)hydrazylthiocarbamic acid and 6-methoxyisoquinoline as starting materials, according to the synthesis method in Example 2.

[0132] 1H NMR (700 MHz, dmso) δ 12.67 (s, 1H), 11.23 (s, 1H), 10.33 (s, 1H), 7.87 (d, J = 7.7 Hz, 1H), 7.59 (d, J = 8.3 Hz, 1H), 7.27 (td, J = 8.3, 1.5Hz, 1H), 7.15 - 7.12 (m, 1H), 6.99 (td, J = 7.7, 1.0 Hz, 1H), 6.68 (dd, J =8.4, 2.2 Hz, 1H), 6.49 (d, J = 2.2 Hz, 1H), 3.86 (s, 3H), 3.81 (s, 3H). Example 26. Synthesis of (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(4-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid (compound 26) The title compound was synthesized from N-(4-(trifluoromethyl)phenyl)hydrazinothiocarbamic acid and 6-methoxyisoquinoline as starting materials, according to the synthesis method in Example 2.

[0133] 1H NMR (700 MHz, dmso) δ 12.80 (s, 1H), 11.25 (s, 1H), 10.89 (s, 1H), 7.93 (d, J = 8.4 Hz, 2H), 7.78 (d, J = 8.5 Hz, 2H), 7.68 (d, J = 8.4 Hz, 1H), 6.70 (dd, J = 8.5, 2.2 Hz, 1H), 6.49 (d, J = 2.2 Hz, 1H), 3.82 (s, 3H). Example 27. Synthesis of (Z)-N-ethyl-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid (compound 27) The title compound was synthesized using N-ethylhydrazylthiocarbamic acid and 6-methoxyisoquinoline as starting materials, according to the synthesis method in Example 2.

[0134] 1H NMR (700 MHz, dmso) δ 12.42 (s, 1H), 11.16 (s, 1H), 9.15 (m, 1H), 7.56 (d, J = 8.4 Hz, 1H), 6.67 (dd, J = 8.4, 2.2 Hz, 1H), 6.47 (d, J = 2.2Hz, 1H), 3.80 (s, 3H), 3.65 - 3.59 (m, 2H), 1.18 (t, J = 7.2 Hz, 3H). Example 28. Synthesis of (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(m-tolyl)hydrazinothiocarbamic acid (compound 28) The title compound was synthesized using N-(3-methylphenyl)hydrazylthiocarbamic acid and 6-methoxyisoquinoline as starting materials, following the synthesis method of Example 2.

[0135] 1H NMR (700 MHz, dmso) δ 12.64 (s, 1H), 11.21 (s, 1H), 10.64 (s, 1H), 7.69 (d, J = 8.4 Hz, 1H), 7.44 - 7.40 (m, 2H), 7.29 (t, J = 7.7 Hz, 1H), 7.08 (d, J = 7.5 Hz, 1H), 6.69 (dd, J = 8.4, 2.1 Hz, 1H), 6.49 (d, J = 2.2 Hz, 1H), 3.81 (s, 3H), 2.33 (s, 3H). Example 29. Synthesis of (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid (compound 29) The title compound was synthesized using phenylhydrazine thiocarbamic acid and 5-methylisoquinoline as starting materials, following the synthesis method of Example 2.

[0136] 1H NMR (700 MHz, dmso) δ 12.77 (s, 1H), 11.15 (s, 1H), 10.81 (s, 1H), 7.64 - 7.58 (m, 3H), 7.43 (t, J = 7.9 Hz, 2H), 7.28 (m, 1H), 7.18 (dd, J =8.0, 0.9 Hz, 1H), 6.83 (d, J = 7.9 Hz, 1H), 2.30 (s, 3H). Example 30. Synthesis of (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid (compound 30) The title compound was synthesized using N-(3-nitrophenyl)hydrazinothiocarbamic acid and 5-methylisoquinoline as starting materials, following the synthesis method of Example 2.

[0137] 1H NMR (700 MHz, dmso) δ 12.94 (s, 1H), 11.21 (s, 1H), 11.08 (s, 1H), 8.65 (t, J = 2.1 Hz, 1H), 8.17 (m, 1H), 8.12 (m, 1H), 7.72 (t, J = 8.1 Hz, 1H), 7.60 (s, 1H), 7.20 (d, J = 7.9 Hz, 1H), 6.84 (d, J = 7.9 Hz, 1H), 2.31 (s, 3H). Example 31. Synthesis of (Z)-N-(2-methoxyphenyl)-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid (compound 31) The title compound was synthesized using N-(2-methoxyphenyl)hydrazinothiocarbamic acid and 5-methylisoxazole as starting materials, according to the synthesis method in Example 2.

[0138] 1H NMR (700 MHz, dmso) δ 12.76 (s, 1H), 11.16 (s, 1H), 10.44 (s, 1H), 7.74 (d, J = 7.8 Hz, 1H), 7.52 (s, 1H), 7.29 (m, 1H), 7.18 (d, J = 7.8 Hz, 1H), 7.14 (d, J = 8.3 Hz, 1H), 7.00 (td, J = 8.0, 1.1 Hz, 1H), 6.84 (d, J =8.0 Hz, 1H), 3.85 (s, 3H), 2.31 (s, 3H). Example 32. Synthesis of (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-(3-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid (compound 32) The title compound was synthesized from N-(3-(trifluoromethyl)phenyl)hydrazinothiocarbamic acid and 5-methylisoxazole as starting materials, according to the synthesis method in Example 2.

[0139] 1H NMR (700 MHz, dmso) δ 12.89 (s, 1H), 11.20 (s, 1H), 10.99 (s, 1H), 7.20-7.17 (m, 2H), 6.95 (m, 1H), 6.88-6.77 (m, 4H), 2.26 (s, 3H). Example 33. Synthesis of (Z)-N-(4-fluorophenyl)-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid (compound 33) The title compound was synthesized using N-(4-fluorophenyl)hydrazinothiocarbamic acid and 5-methylisoxazole as starting materials, according to the synthesis method in Example 2.

[0140] 1H NMR (700 MHz, dmso) δ 12.78 (s, 1H), 11.16 (s, 1H), 10.82 (s, 1H), 7.60-7.57 (m, 3H), 7.26 (m, 2H), 7.18 (dd, J = 7.9, 0.9 Hz, 1H), 6.83 (d, J =7.9 Hz, 1H), 2.30 (s, 3H). Example 34. Synthesis of (Z)-2-(5-bromo-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid (compound 34) The title compound was synthesized using thiocysteine ​​and 5-bromoisoxazole as starting materials, according to the synthesis method in Example 2.

[0141] 1H NMR (500 MHz, dmso) δ 12.28 (s, 1H), 11.28 (s, 1H), 9.10 (s, 1H), 8.81 (s, 1H), 7.88 (d, J = 2.1 Hz, 1H), 7.51 - 7.48 (m, 1H), 6.90 - 6.87 (m, 1H). Example 35. Synthesis of (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid (compound 35) The title compound was synthesized from N-(3-nitrophenyl)hydrazinothiocarbamic acid and 4-bromoisoflavone using the synthesis method of Example 2.

[0142] 1H NMR (500 MHz, dmso) δ 13.23 (s, 1H), 11.54 (s, 1H), 10.33 (s, 1H), 8.86 (s, 1H), 8.19 - 7.98 (m, 2H), 7.71 (t, J = 8.2 Hz, 1H), 7.36 - 7.23 (m, 2H), 6.97 (m, 1H). Example 36. Synthesis of (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid (compound 36) The title compound was synthesized from N-(3-nitrophenyl)hydrazinothiocarbamic acid and 6-bromoisoflavone as starting materials, according to the synthesis method in Example 2.

[0143] 1H NMR (500 MHz, dmso) δ 13.23 (s, 1H), 11.54 (s, 1H), 10.33 (s, 1H), 8.86 (t, J = 2.1 Hz, 1H), 8.19 - 7.98 (m, 2H), 7.71 (t, J = 8.2 Hz, 1H), 7.36- 7.23 (m, 2H), 6.97 (m, 1H). Example 37. Synthesis of (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid (compound 37) The title compound was synthesized from N-(4-nitrophenyl)hydrazinothiocarbamic acid and 4-bromoisoflavone using the synthesis method of Example 2.

[0144] 1H NMR (500 MHz, dmso) δ 13.22 (s, 1H), 11.55 (s, 1H), 10.47 (s, 1H), 8.31 - 8.26 (m, 2H), 8.17 - 8.12 (m, 2H), 7.34 - 7.27 (m, 2H), 6.98 (m, 1H). Example 38. Synthesis of (Z)-2-(5-bromo-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid (compound 38) The title compound was synthesized using N-(4-nitrophenyl)hydrazinothiocarbamic acid and 5-bromoisoflavin as starting materials, according to the synthesis method in Example 2.

[0145] 1H NMR (500 MHz, dmso) δ 12.81 (s, 1H), 11.39 (s, 1H), 11.11 (s, 1H), 8.34 - 8.28 (m, 2H), 8.10 - 8.06 (m, 2H), 7.99 (d, J = 2.1 Hz, 1H), 7.54 (m, 1H), 6.91 (m, 1H). Example 39. Synthesis of (Z)-N-(2-methoxyphenyl)-2-(2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid (compound 39) The title compound was synthesized using N-(2-methoxyphenyl)hydrazine thiocarbamic acid and isoquinolinone as starting materials, according to the synthesis method in Example 2.

[0146] 1H NMR (500 MHz, dmso) δ 12.79 (s, 1H), 11.25 (s, 1H), 10.43 (s, 1H), 7.86 (dd, J = 7.8, 1.2 Hz, 1H), 7.68 (d, J = 7.5 Hz, 1H), 7.38 (m, 1H), 7.29 (ddd, J = 8.2, 7.5, 1.7 Hz, 1H), 7.16 - 7.09 (m, 2H), 7.00 (td, J = 7.7, 1.2Hz, 1H), 6.95 (m, 1H), 3.87 (s, 3H). Example 40. Synthesis of (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid (compound 40) The title compound was synthesized using thiocysteine ​​and 4-bromoisoflavin as starting materials, according to the synthesis method in Example 2.

[0147] 1H NMR (500 MHz, dmso) δ 12.77 (s, 1H), 11.43 (s, 1H), 9.30 (s, 1H), 7.71 (s, 1H), 7.29 - 7.23 (m, 2H), 6.94 (m, 1H). Example 41. Synthesis of (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid (compound 41) The title compound was synthesized using thiocysteine ​​and 6-bromoisoflavin as starting materials, according to the synthesis method in Example 2.

[0148] 1H NMR (500 MHz, dmso) δ 12.37 (s, 1H), 11.31 (s, 1H), 9.08 (s, 1H), 8.73 (s, 1H), 7.58 (d, J = 8.1 Hz, 1H), 7.30 (m, 1H), 7.08 (d, J = 1.7 Hz, 1H). Example 42. Synthesis of (Z)-2-(7-bromo-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid (compound 42) The title compound was synthesized using thiocysteine ​​and 7-bromoisoflavin as starting materials, according to the synthesis method in Example 2.

[0149] 1H NMR (500 MHz, dmso) δ 12.40 (s, 1H), 11.50 (s, 1H), 9.11 (s, 1H), 8.76 (s, 1H), 7.66 (d, J = 7.3 Hz, 1H), 7.56 - 7.52 (m, 1H), 7.04 (dt, J =7.0, 6.0 Hz, 1H). Example 43. Synthesis of (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid (compound 43) The title compound was synthesized using phenylhydrazine thiocarbamate and 4-bromoisoflavin as starting materials, according to the synthesis method in Example 2.

[0150] 1H NMR (500 MHz, dmso) δ 13.04 (s, 1H), 11.50 (s, 1H), 9.98 (s, 1H), 7.75-7.71 (m, 2H), 7.45-7.40 (m, 2H), 7.32 - 7.24 (m, 3H), 6.97 (m, 1H). Example 44. Synthesis of (Z)-2-(5-bromo-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid (compound 44) The title compound was synthesized using phenylhydrazine thiocarbamate and 5-bromoisoflavin as starting materials, according to the synthesis method in Example 2.

[0151] 1H NMR (500 MHz, dmso) δ 12.61 (s, 1H), 11.34 (s, 1H), 10.89 (s, 1H), 8.01 (d, J = 2.1 Hz, 1H), 7.62 - 7.59 (m, 2H), 7.54 - 7.51 (m, 1H), 7.46 -7.42 (m, 2H), 7.31 - 7.27 (m, 1H), 6.90 (m, 1H). Example 45. Synthesis of (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid (compound 45) The title compound was synthesized using phenylhydrazine thiocarbamate and 6-bromoisoflavin as starting materials, following the synthesis method of Example 2.

[0152] 1H NMR (500 MHz, dmso) δ 12.69 (s, 1H), 11.37 (s, 1H), 10.86 (s, 1H), 7.71 (d, J = 8.1 Hz, 1H), 7.62-7.60 (m, 2H), 7.45 - 7.40 (m, 2H), 7.33 (dd, J = 8.1, 2.1 Hz, 1H), 7.28 (ddt, J = 5.4, 4.3, 2.1 Hz, 1H), 7.10 (d, J = 1.6Hz, 1H). Example 46. Synthesis of (Z)-2-(7-bromo-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid (compound 46) The title compound was synthesized using phenylhydrazine thiocarbamate and 7-bromoisoflavin as starting materials, following the synthesis method of Example 2.

[0153] 1H NMR (500 MHz, dmso) δ 12.73 (s, 1H), 11.56 (s, 1H), 10.89 (s, 1H), 7.7 (m, 1H), 7.62-7.58 (m, 2H), 7.57 (m, 1H), 7.46 - 7.41 (m, 2H), 7.29 (m, 1H), 7.07 (m, 1H). Example 47. Synthesis of (Z)-N-(3-nitrophenyl)-2-(2-oxoindoline-3-ylidene)hydrazinothiocarbamic acid (compound 47) The title compound was synthesized using N-(3-nitrophenyl)hydrazinothiocarbamic acid and isoflavone as starting materials, following the synthesis method of Example 2.

[0154] 1H NMR (500 MHz, dmso) δ 12.95 (s, 1H), 11.29 (s, 1H), 11.09 (s, 1H), 8.66 (t, J = 2.2 Hz, 1H), 8.18 (m, 1H), 8.12 (m, 1H), 7.77 (m, 1H), 7.72 (dd, J = 10.8, 5.5 Hz, 1H), 7.39 (m, 1H), 7.13 (m, 1H), 6.96 (m, 1H). Example 48. Synthesis of (Z)-2-(5-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid (compound 48) The title compound was synthesized from N-(3-nitrophenyl)hydrazinothiocarbamic acid and 5-bromoisoflavone using the synthesis method of Example 2.

[0155] 1H NMR (500MHz, DMSO) δ12.76 (s, 1H), 11.39 (s, 1H), 11.10 (s, 1H), 8.65 (m, 1H), 8.17 ( m, 1H), 8.13 (m, 1H), 7.98 (d, J=2.1Hz, 1H), 7.74 (m, 1H), 7.54 (m, 1H), 6.92 (m, 1H). Example 49. Synthesis of (Z)-2-(7-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid (compound 49) The title compound was synthesized from N-(3-nitrophenyl)hydrazinoaminothioacetamide and 7-bromoisoflavone using the synthesis method of Example 2.

[0156] 1H NMR (500MHz, dmso) δ12.88 (s, 1H), 11.60 (s, 1H), 11.15 (s, 1H), 8.65 (t, J=2.2Hz, 1H), 8.18 (m, 1H) , 8.12 (m, 1H), 7.79 (dd, J=9.2, 4.9Hz, 1H), 7.73 (dd, J=10.9, 5.4Hz, 1H), 7.59 (m, 1H), 7.10 (m, 1H). Example 50. Synthesis of (Z)-N-(3-methoxyphenyl)-2-(2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid (compound 50) The title compound was synthesized from N-(3-methoxyphenyl)hydrazine thiocarbamic acid and isoflavones using the synthesis method of Example 2.

[0157] 1H NMR (500MHz, dmso) δ12.80 (s, 1H), 11.25 (s, 1H), 10.76 (s, 1H), 7.79 (m, 1H), 7.37 (td, J=7.7, 1.3Hz, 1H), 7.34 (s, 1H) , 7.30 (dd, J=4.4, 2.2Hz, 1H), 7.24 (m, 1H), 7.11 (m, 1H), 6.95 (m, 1H), 6.85 (ddd, J=8.2, 2.6, 0.9Hz, 1H), 3.78 (s, 3H). Example 51. Synthesis of (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(3-methoxyphenyl)hydrazylthiocarbamic acid (compound 51) The title compound was synthesized from N-(3-methoxyphenyl)hydrazine thiocarbamic acid and 4-bromoisoindoline using the synthesis method of Example 2.

[0158] 1H NMR (500 MHz, dmso) δ 13.02 (s, 1H), 11.50 (s, 1H), 9.96 (s, 1H), 7.54 (t, J = 2.2 Hz, 1H), 7.32 (m, 1H), 7.30 - 7.25 (m, 2H), 7.22 (m, 1H), 6.96 (m, 1H), 6.84 (ddd, J = 8.3, 2.5, 0.9 Hz, 1H), 3.78 (s, 3H). Example 52. Synthesis of (Z)-2-(5-bromo-2-oxoindoline-3-ylidene)-N-(3-methoxyphenyl)hydrazylthiocarbamic acid (compound 52) The title compound was synthesized from N-(3-methoxyphenyl)hydrazine thiocarbamic acid and 5-bromoisoindoline according to the synthesis method of Example 2.

[0159] 1H NMR (500 MHz, dmso) δ 12.62 (s, 1H), 11.35 (s, 1H), 10.84 (s, 1H), 8.02 (d, J = 2.1 Hz, 1H), 7.52 (m, 1H), 7.34 (m, 1H), 7.28 (t, J = 2.2 Hz, 1H), 7.22 (m, 1H), 6.90 (m, 1H), 6.87 (m, 1H), 3.79 (s, 3H). Example 53. Synthesis of (Z)-N-(4-methoxyphenyl)-2-(2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid (compound 53) The title compound was synthesized from N-(4-methoxyphenyl)hydrazine thiocarbamic acid and isoindoline according to the synthesis method of Example 2.

[0160] 1H NMR (500 MHz, dmso) δ 12.75 (s, 1H), 11.24 (s, 1H), 10.72 (s, 1H), 7.77 (d, J = 7.5 Hz, 1H), 7.48 - 7.45 (m, 2H), 7.37 (td, J = 7.7, 1.3 Hz, 1H), 7.11 (m, 1H), 7.00 - 6.96 (m, 2H), 6.94 (dt, J = 7.9, 0.8 Hz, 1H), 3.78 (s, 3H). Example 54. Synthesis of (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(4-methoxyphenyl)hydrazylthiocarbamic acid (compound 54) The title compound was synthesized from N-(4-methoxyphenyl)hydrazine thiocarbamic acid and 4-bromoisoindoline using the synthesis method of Example 2.

[0161] 1H NMR (500 MHz, dmso) δ 13.02 (s, 1H), 11.49 (s, 1H), 9.85 (s, 1H), 7.54 (m, 2H), 7.29 (m, 2H), 6.98 (m, 3H), 3.78 (s, 3H). Example 55. Synthesis of (Z)-2-(5-bromo-2-oxoindoline-3-ylidene)-N-(4-methoxyphenyl)hydrazylthiocarbamic acid (compound 55) The title compound was synthesized from N-(4-methoxyphenyl)hydrazine thiocarbamic acid and 5-bromoisosorbic acid according to the synthesis method in Example 2.

[0162] 1H NMR (500 MHz, dmso) δ 12.56 (s, 1H), 11.33 (s, 1H), 10.80 (s, 1H), 8.00 (d, J = 1.9 Hz, 1H), 7.51 (m, 1H), 7.48 - 7.43 (m, 2H), 7.01 - 6.97 (m, 2H), 6.90 (m, 1H), 3.79 (s, 3H). Example 56. Synthesis of N-(3,4-dimethoxyphenyl)hydrazinothiocarbamic acid (compound 56) The title compound was synthesized using 3,4-dimethoxyphenyl isothiocyanate and hydrazine as starting materials, according to the synthesis method in Example 2.

[0163] 1H NMR (700 MHz, dmso) δ 9.53 (s, 1H), 9.00 (s, 1H), 7.36 (s, 1H), 7.10 (s, 1H), 6.87 (d, J = 8.6 Hz, 1H), 4.73 (s, 2H), 3.72 (s, 3H), 3.71 (s, 3H). Example 57. Synthesis of (Z)-N-(4-fluorophenyl)-2-(2-oxoindoline-3-ylidene)hydrazinothiocarbamic acid (compound 57) The title compound was synthesized using N-(4-fluorophenyl)hydrazinothiocarbamic acid and isosorbide as starting materials, according to the synthesis method in Example 2.

[0164] 1H NMR (700 MHz, dmso) δ 12.80 (s, 1H), 11.27 (s, 1H), 10.84 (s, 1H), 7.75 (d, J = 7.5 Hz, 1H), 7.62 - 7.57 (m, 2H), 7.37 (td, J = 7.7, 1.2 Hz, 1H), 7.29 - 7.24 (m, 2H), 7.11 (td, J = 7.5, 0.7 Hz, 1H), 6.95 (d, J = 7.8Hz, 1H). Example 58. Synthesis of (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid (compound 58) The title compound was synthesized using N-(4-nitrophenyl)hydrazinothiocarbamic acid and 6-bromoisoflavin as starting materials, according to the synthesis method in Example 2.

[0165] 1H NMR (700MHz, DMSO) δ12.89 (s, 1H), 11.43 (s, 1H), 11.15 (s, 1H), 8.36-8.22 (m, 2H), 8.15-8.0 0 (m, 2H), 7.70 (d, J=8.1Hz, 1H), 7.32 (ddd, J=21.7, 11.6, 4.4Hz, 1H), 7.12 (d, J=1.6Hz, 1H). Example 60. Synthesis of (Z)-2-(6-chloro-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid (compound 60) The title compound was synthesized using phenylhydrazine thiocarbamate and 6-chloroisoflavin as starting materials according to the synthesis method in Example 2.

[0166] 1H NMR (700MHz, DMSO) δ12.68 (s, 1H), 11.40 (s, 1H), 10.87 (s, 1H), 7.77 (d, J=8.1Hz, 1H), 7.60 (d, J=7.7Hz , 2H), 7.43 (t, J=7.9Hz, 2H), 7.28 (t, J=7.4Hz, 1H), 7.19 (dd, J=8.1, 1.8Hz, 1H), 6.98 (d, J=1.8Hz, 1H). Example 61. Synthesis of (Z)-2-(4-chloro-2-oxoindoline-3-ylidene)-N-(2-methoxyphenyl)hydrazylthiocarbamic acid (compound 61) The title compound was synthesized using N-(2-methoxyphenyl)hydrazine thiocarbamate and 4-chloroisoflavin as starting materials, according to the synthesis method in Example 2.

[0167] 1H NMR (700MHz, DMSO) δ12.96 (s, 1H), 11.53 (s, 1H), 10.35 (s, 1H), 8.83 (d, J=7.7Hz, 1H), 7.37 (t, J=8.0Hz, 1H), 7.20 (m, 1H), 7.16-7.13 (m, 2H), 7.01 (m, 1H), 6.92 (d, J=7.8Hz, 1H), 3.88 (s, 3H). Example 62. Synthesis of (Z)-2-(5-fluoro-2-oxoindoline-3-ylidene)-N-(3-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid (compound 62) The title compound was synthesized from N-(3-(trifluoromethyl)phenyl)hydrazinothiocarbamic acid and 5-fluoroisosorbic acid as starting materials, according to the synthesis method in Example 2.

[0168] 1H NMR (700 MHz, dmso) δ 12.78 (s, 1H), 11.29 (s, 1H), 11.00 (s, 1H), 8.15 - 7.91 (m, 2H), 7.76 - 7.55 (m, 3H), 7.23 (m, 1H), 6.95 (dd, J = 8.6, 4.1 Hz, 1H). Example 63. Synthesis of (Z)-N-benzyl-2-(5-fluoro-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid (compound 63) The title compound was synthesized using N-benzylhydrazine thiocarbamic acid and 5-fluoroisosorbic acid as starting materials, according to the synthesis method in Example 2.

[0169] 1H NMR (700 MHz, dmso) δ 12.52 (s, 1H), 11.20 (s, 1H), 9.85 (t, J = 6.2 Hz, 1H), 7.45 (dd, J = 8.1, 2.6 Hz, 1H), 7.36 - 7.30 (m, 4H), 7.24 (dd, J = 9.5, 4.3 Hz, 1H), 7.16 (td, J = 9.4, 2.7 Hz, 1H), 6.90 (dd, J = 8.6, 4.1Hz, 1H), 4.86 (d, J = 6.2 Hz, 2H). Example 64. Synthesis of (Z)-2-(5-fluoro-2-oxoindoline-3-ylidene)-N-(m-tolyl)hydrazylthiocarbamic acid (compound 64).

[0170] The title compound was synthesized from N-(3-methylphenyl)hydrazinothiocarbamic acid and 5-fluoroisosorbic acid as starting materials, according to the synthesis method in Example 2.

[0171] 1H NMR (700 MHz, dmso) δ 12.65 (s, 1H), 11.26 (s, 1H), 10.79 (s, 1H), 7.65 (dd, J = 8.1, 2.5 Hz, 1H), 7.46 - 7.36 (m, 2H), 7.32 (t, J = 7.7 Hz, 1H), 7.21 (td, J = 9.3, 2.7 Hz, 1H), 7.10 (d, J = 7.5 Hz, 1H), 6.94 (dd, J = 8.6, 4.1 Hz, 1H), 2.34 (s, 3H). Example 65. Synthesis of (Z)-2-(7-fluoro-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid (compound 65) The title compound was synthesized from N-(4-nitrophenyl)hydrazinothiocarbamic acid and 7-fluoroisosorbic acid using the synthesis method of Example 2.

[0172] 1H NMR (700 MHz, dmso) δ 12.96 (s, 1H), 11.84 (s, 1H), 11.17 (s, 1H), 8.37 - 8.20 (d, J = 9.1 Hz, 2H), 8.07 (d, J = 9.1 Hz, 2H), 7.63 (d, J = 7.5Hz, 1H), 7.32 (dd, J = 9.9, 8.9 Hz, 1H), 7.14 (m, 1H). Example 66. Synthesis of (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(3-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid (compound 66) The title compound was synthesized from N-(3-(trifluoromethyl)phenyl)hydrazinothiocarbamic acid and 6-methoxyisoquinoline as starting materials, according to the synthesis method in Example 2.

[0173] 1H NMR (700 MHz, dmso) δ 12.77 (s, 1H), 11.25 (s, 1H), 10.87 (s, 1H), 8.06 (s, 1H), 8.00 (d, J = 8.0 Hz, 1H), 7.69 - 7.58 (m, 3H), 6.68 (m, 1H), 6.47 (d, 2.2 Hz, 1H), 3.82 (s, 3H). Example 67. Synthesis of (Z)-N-(4-fluorophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid (compound 67) The title compound was synthesized using N-(4-fluorophenyl)hydrazylthiocarbamate and 6-methoxyisoquinoline as starting materials, according to the synthesis method in Example 2.

[0174] 1H NMR (700 MHz, dmso) δ 12.68 (s, 1H), 11.23 (s, 1H), 10.71 (s, 1H), 7.66 (d, J = 8.4 Hz, 1H), 7.62 - 7.55 (m, 2H), 7.29 - 7.21 (m, 2H), 6.67 (ddd, J = 34.4, 8.6, 2.3 Hz, 1H), 6.46 (d, J = 2.3 Hz, 1H), 3.81 (s, 3H). Example 68. Synthesis of (Z)-N-(4-bromophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid (compound 68) The title compound was synthesized using N-(4-bromophenyl)hydrazinothiocarbamic acid and 6-methoxyisoquinoline as starting materials, according to the synthesis method in Example 2.

[0175] 1H NMR (700 MHz, dmso) δ 12.72 (s, 1H), 11.23 (s, 1H), 10.72 (s, 1H), 7.67 (d, J = 8.4 Hz, 1H), 7.60 (m, 4H), 6.68 (m, 1H), 6.46 (d, J = 2.3 Hz, 1H), 3.82 (s, 3H). Example 69. Synthesis of (Z)-N-benzyl-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid (compound 69) The title compound was synthesized from N-benzylhydrazine thiocarbamic acid and 6-methoxyisoquinoline according to the synthesis method of Example 2.

[0176] 1H NMR (700 MHz, dmso) δ 12.53 (s, 1H), 11.18 (s, 1H), 9.69 (t, J = 6.2 Hz, 1H), 7.55 (d, J = 8.4 Hz, 1H), 7.37-7.32 (m, 4H), 7.26 (t, J = 7.1Hz, 1H), 6.63 (m, 1H), 6.45 (d, J = 2.3 Hz, 1H), 4.86 (d, J = 6.5 Hz, 2H), 3.81 (s, 3H). Example 70. Synthesis of (Z)-N-(4-bromophenyl)-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid (compound 70) The title compound was synthesized from N-(4-bromophenyl)hydrazinothiocarbamic acid and 5-methylisoxazole according to the synthesis method in Example 2.

[0177] 1H NMR (700 MHz, dmso) δ 12.82 (s, 1H), 11.18 (s, 1H), 10.84 (s, 1H), 7.71 - 7.51 (m, 5H), 7.19 (dd, J = 8.0 Hz, 1H), 6.82 (d, J = 8.0 Hz, 1H), 2.28 (s, 3H). Example 71. Synthesis of (Z)-N-(4-cyanophenyl)-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid (compound 71) The title compound was synthesized using N-(4-cyanophenyl)hydrazinothiocarbamic acid and 5-methylisoxazole as starting materials, according to the synthesis method in Example 2.

[0178] 1H NMR (700 MHz, dmso) δ 12.94 (s, 1H), 11.19 (s, 1H), 10.96 (s, 1H), 7.97 (d, J = 8.6 Hz, 2H), 7.89 (m, J = 8.6 Hz, 2H), 7.60 (s, 1H), 7.20 (d, J = 7.9 Hz, 1H), 6.82 (d, J = 7.9 Hz, 1H), 2.28 (s, 3H). Example 72. Synthesis of (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid (compound 72) The title compound was synthesized from phenylhydrazine thiocarbamic acid and 5-methylisoxazole according to the synthesis method of Example 2.

[0179] 1H NMR (700 MHz, dmso) δ 12.91 (s, 1H), 11.19 (s, 1H), 10.99 (s, 1H), 7.93 (d, J = 8.4 Hz, 2H), 7.79 (d, J = 8.4 Hz, 2H), 7.61 (s, 1H), 7.17 (m, 1H), 6.84 (d, J = 7.9 Hz, 1H), 2.31 (s, 3H). Example 73. Synthesis of (Z)-N-ethyl-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid (compound 73) The title compound was synthesized from N-ethylhydrazine thiocarbamic acid and 5-methylisoxazole according to the synthesis method of Example 2.

[0180] 1H NMR (700 MHz, dmso) δ 12.52 (s, 1H), 11.10 (s, 1H), 9.27 (t, J =5.8 Hz, 1H), 7.49 (s, 1H), 7.19 (m, 1H), 6.81 (m, 1H), 3.73 - 3.53 (m, 2H), 2.31 (s, 3H), 1.19 (t, J = 7.2 Hz, 3H). Example 74. Synthesis of (Z)-N-benzyl-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid (compound 74) The title compound was synthesized from N-benzylhydrazine thiocarbamic acid and 5-methylisoxazole according to the synthesis method of Example 2.

[0181] 1H NMR (700 MHz, dmso) δ 12.64 (s, 1H), 11.11 (s, 1H), 9.81 (t, J =6.3 Hz, 1H), 7.48 (s, 1H), 7.35 (m, 4H), 7.25 (dd, J = 12.6, 12.4 Hz, 1H), 7.16 (dd, J = 7.9, 0.9 Hz, 1H), 6.82 (d, J = 7.9 Hz, 1H), 4.87 (d, J = 8.7Hz, 2H), 2.28 (s, 3H). Example 75. Synthesis of (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-(m-tolyl)hydrazinothiocarbamic acid (compound 75) The title compound was synthesized using N-(3-methylphenyl)hydrazinothiocarbamic acid and 5-methylisoxazole as starting materials, following the synthetic method of Example 2.

[0182] 1H NMR (700 MHz, dmso) δ 12.75 (s, 1H), 11.15 (s, 1H), 10.75 (s, 1H), 7.62 (s, 1H), 7.48 - 7.38 (m, 2H), 7.30 (t, J = 7.7 Hz, 1H), 7.18 (dd, J = 7.9, 0.8 Hz, 1H), 7.09 (d, J = 7.5 Hz, 1H), 6.83 (d, J = 7.9 Hz, 1H), 2.34 (s, 3H), 2.30 (s, 3H). Example 76. Synthesis of (Z)-2-(2-oxo-5-(trifluoromethoxy)indoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid (compound 76) The title compound was synthesized using the synthetic method of Example 2, starting with phenylhydrazine thiocarbamic acid and 5-(trifluoromethoxy)isoxazole.

[0183] 1H NMR (700 MHz, dmso) δ 12.65 (s, 1H), 11.42 (s, 1H), 10.90 (s, 1H), 7.80 (s, 1H), 7.59 (d, J = 7.7 Hz, 2H), 7.44 (t, J = 7.7 Hz, 2H), 7.37 (dd, J = 8.5, 1.9 Hz, 1H), 7.30 (t, J = 7.4 Hz, 1H), 7.03 (d, J = 8.5 Hz, 1H). Example 77. Synthesis of (Z)-N-(3-nitrophenyl)-2-(2-oxo-5-(trifluoromethoxy)indoline-3-ylidene)hydrazylthiocarbamic acid (compound 77) The title compound was synthesized using N-(3-nitrophenyl)hydrazinothiocarbamic acid and 5-(trifluoromethoxy)isoxazole as starting materials, following the synthetic method of Example 2.

[0184] 1H NMR (700 MHz, dmso) δ 12.81 (s, 1H), 11.47 (s, 1H), 11.14 (s, 1H), 8.63 (t, J = 2.1 Hz, 1H), 8.15 (m, 2H), 7.77 (s, 1H), 7.73 (t, J = 8.2 Hz, 1H), 7.40 (dd, J = 8.5, 1.9 Hz, 1H), 7.04 (d, J = 8.5 Hz, 1H). Example 78. Synthesis of (Z)-N-(4-fluorophenyl)-2-(2-oxo-5-(trifluoromethoxy)indoline-3-ylidene)hydrazylthiocarbamic acid (compound 78) The title compound was synthesized using N-(4-fluorophenyl)hydrazinothiocarbamic acid and 5-(trifluoromethoxy)isoxazole as starting materials, following the synthetic method of Example 2.

[0185] 1H NMR (700 MHz, dmso) δ 12.66 (s, 1H), 11.43 (s, 1H), 10.90 (s, 1H), 7.78 (s, 1H), 7.60 - 7.56 (m, 2H), 7.38 (d, J = 10.1 Hz, 1H), 7.30 - 7.26 (m, 2H), 7.04 (d, J = 8.5 Hz, 1H). Example 79. Synthesis of (Z)-N-(4-bromophenyl)-2-(2-oxo-5-(trifluoromethoxy)indoline-3-ylidene)hydrazylthiocarbamic acid (compound 79) The title compound was synthesized from N-(4-bromophenyl)hydrazinothiocarbamic acid and 5-(trifluoromethoxy)isoindoline as starting materials, according to the synthesis method in Example 2.

[0186] 1H NMR (700 MHz, dmso) δ 12.70 (s, 1H), 11.43 (s, 1H), 10.91 (s, 1H), 7.78 (s, 1H), 7.63 (d, J = 8.8 Hz, 2H), 7.59 (d, J = 8.8 Hz, 2H), 7.38 (dd, J= 8.5, 1.8 Hz, 1H), 7.03 (d, J= 8.5 Hz, 1H). Table 1 below lists the compounds of Formula 1 above.

[0187] (Table 1)

[0188] Example Experimental Example 1. Evaluation of the DNA binding inhibition effect of EGR-1 using EDBA In this experiment, the activity of the compounds of the present invention in inhibiting EGR-1 binding was evaluated using EDBA (EGR-1-DNA binding activity assay). Figure 1 The EDBA experimental method is briefly shown.

[0189] (1) Preparation of biotin-labeled EGR1 binding sequence (Biotin-EBS) oligonucleotides.

[0190] Single-stranded sense and antisense oligonucleotides with the EGR1-binding sequence (EBS) shown below were prepared by Bioneer in Daejeon, South Korea. Biotin was labeled at the 5' end of the sense oligonucleotide. The sense and antisense oligonucleotides were heated at 95°C for 5 minutes and then left to stand at room temperature overnight to allow them to bind. Hereinafter, the bound sense and antisense oligonucleotides will be referred to as Biotin-EBS.

[0191] sense sequence: biotin-5'-TCGCCCCCGCTCGCCCCCGCTGGATCC-3' Antonym sequence: 5'-GGATCCAGCGGGGGCGAGCGGGGGCGA-3' (2) Preparation of biotin-EBS oligonucleotide coated plates Biotin-EBS oligonucleotides were added at a concentration of 0.5 pmol / 50 μL / well to a 96-well plate coated with avidin (Thermo Fisher Scientific) and incubated at 37°C for 1 hour and 30 minutes to allow the biotin-EBS oligonucleotides to adhere to the plate. The plate was then washed three times with phosphate-buffered saline (PBS / Tween20) containing 0.05% Tween 20 to remove any unattached biotin-EBS oligonucleotides. Finally, 200 μL of PBS / Tween20 solution containing 2% bovine serum albumin (BSA) was added to each well, and the plate was incubated at 37°C for 1 hour to block any oligonucleotides not attached to the plate surface.

[0192] (3) Analysis of EGR-1 binding inhibition ability Add 50 μL of the solution containing the sample compound reacting with EGR-1 protein to the biotin-EBS oligonucleotide plating plate prepared in (2) above. At this time, the compound is screened based on the following principle: if the sample compound does not bind to EGR-1 protein, the EGR-1 antibody reaction will cause a color reaction; but if the sample compound binds to EGR-1 protein and inhibits the DNA binding ability of EGR-1, no color reaction will occur.

[0193] One μL of the compound sample was reacted with 49 μL of EGR-1 overexpressing cell lysate containing 10 mM Tris, pH 7.5, 50 mM KCl, 2% BSA, and 8% glycerol at room temperature for 30 minutes. The mixture was then added to wells coated with biotin-EBS and reacted for another hour at room temperature. After washing three times with PBS / Tween 20 solution, 50 μL of EGR-1 antibody was added to each well, and the mixture was reacted for 1 hour at room temperature. After washing three times with PBS / Tween 20 solution, a secondary antibody conjugated with horseradish peroxidase (goat anti-rabbit IgG antibody, Cell Signaling Technology, Danfoss, MD, USA) was diluted 1:1000, and the mixture was reacted for 1 hour at room temperature. After washing five times with PBS / Tween 20 solution, 50 μL of the substrate tetramethylbenzidine was added to each well. After reacting for 5 minutes, 100 μL of 2N sulfuric acid solution was added to terminate the reaction. The DNA binding inhibition rate was determined using the following formula 1.

[0194] (Mathematical Formula 1) EGR-1 DNA binding inhibition rate (%) = 100 - {(As / Ap) × 100} As: Absorbance of the sample after the reaction of EGR-1 overexpression cell lysate with the compound. Ap: Absorbance of samples treated only with EGR-1 overexpression cell lysates Table 2 shows the ability of the compounds prepared in the above examples to inhibit EGR-1 DNA binding using EDBA.

[0195] (Table 2)

[0196] Figure 2 The results of EGR-1 DNA binding inhibition rate determination for compounds of Formula 1 (79 in total) are shown. Among the 79 compounds of Formula 1, compounds 21, 23, and 25 exhibited the best EGR-1 DNA binding inhibition activity.

[0197] Experimental Example 2: In vitro cytotoxicity analysis of the compounds of the present invention This experiment analyzed the in vitro cytotoxicity of compounds of Formula 1. Specifically, for the 79 compounds represented by Formula 1, cell viability was determined in HaCaT keratinocytes using the CCK-8 assay kit (Sigma-Aldrich), and compounds with cell viability lower than 80% of the solvent-treated control group were considered toxic. 1×10 4 HaCaT keratinocytes were seeded in 96-well cell culture plates, treated with each compound at a concentration of 100 μM, and incubated for 24 hours, followed by the addition of 10 μL of CCK-8 solution. Two hours later, the absorbance of the cell culture was measured at a wavelength of 450 nm, and cell viability was calculated using Equation 2 below.

[0198] [Mathematical Expression 2] Cell viability (%) = (As / Av) × 100 As: Absorbance of the sample treated with the compound Av: Absorbance of the sample treated with solvent only In the experiments, the survival rate of the control cell population treated with 0.1% dimethyl sulfoxide (DMSO) was considered 100%. When this value was considered 100%, the survival rates of all 79 compounds of Formula 1 of the present invention were measured to be above 90%. These results indicate that the compounds of Formula 1 of the present invention are non-toxic to human skin HaCaT keratinocytes at a concentration of 100 μM.

[0199] Experimental Example 3: Prediction of EGR-1 binding sites of the compounds of this invention In this experiment, molecular docking was used to predict the EGR-1 binding site of the compound of the present invention.

[0200] To determine the binding between EGR-1 and the compound, molecular docking experiments were performed on a computer using the Autodock vina program [J. Comp. Chem.31: 455 (2010)] and the Sybyl program (Tripos, St. Louis, MO).

[0201] The tertiary structure of EGR-1 has been registered in multiple protein data banks (PDBs). One of them, 5n14.pdb, is a structure determined by nuclear magnetic resonance spectroscopy and contains only 25 amino acid residues. 4x9j.pdb and 4r2a.pdb are structures determined by X-ray crystallography. Because 4r2a.pdb contains more amino acid residues than 4x9j.pdb, this experiment uses 4r2a.pdb [Genes Dev. 28:2304 (2014)]. The 4r2a.pdb structure contains amino acid residues between E335 and D423, covering ZnF1 (338-362), ZnF2 (368-390), and ZnF3 (396-418) between E335 and D423. Since 4r2a.pdb does not contain ligands, the MOLCAD module included in the Sybyl program was used to determine the binding sites (F377, S378, H382, T385, H386, T389, R407). Using the GRIDbox module of the AutoDockTools program [J. Comp. Chem. 31: 455 (2010)], the docking box size was determined to be 24 x, y, and z, with the center coordinates of the docking box being x=-8.194, y=-4.889, and z=-6.333. Subsequently, docking experiments were performed on a computer using the Autodock vina program.

[0202] The docking results were analyzed using the LigPlot program provided by the European Bioinformatics Institute [Protein. 1999; 37:228], and three-dimensional structural images were generated using the PyMOL program (The PyMOL Molecular Graphics System, Version 1.0r1, Schrödinger, LLC). To obtain the apo protein of EGR-1, oligonucleotides were removed from 4r2a.pdb using the Sybyl program.

[0203] The three compounds (compounds 21, 23, and 25) with the highest EGR-1 DNA binding inhibition rate in Experiment 1 were selected, and their binding states with EGR-1 were analyzed. The three-dimensional structures of compounds 21, 23, and 25 were constructed using the template 1-[(7-fluoro-2-hydroxy-1H-indol-3-yl)imino]-3-(4-fluorophenyl)thiourea (PubChem CID=136269038) registered in PubChem (https: / / pubchem.ncbi.nlm.nih.gov / ) and the PyMOL program. Subsequently, the energy was minimized using the UCSFChimera program to obtain the three-dimensional structure with the lowest energy [JComput Chem. 2004 Oct; 25(13):1605].

[0204] Nine docking experiments were repeated using the AutoDock vina program, yielding nine complexes of compound 21 bound to EGR-1. The binding energies of these nine complexes ranged from -6.9 kcal / mol to -5.7 kcal / mol, thus indicating stable binding. Among these nine complexes, the one with the lowest binding energy exhibited a stable conformation of compound 21 bound to EGR-1, and was therefore identified as the result of the binding of compound 21 to EGR-1. The interaction between the amino acid residues of compound 21 and EGR-1 was analyzed using the LigPlot program.

[0205] Figure 4 This demonstrates the interaction between compound 21 and the amino acid residues of EGR-1. (See reference) Figure 4 Compound 21 exhibits hydrophobic interactions with I361, F377, and H382, and forms hydrogen bonds with R357, S378, and R379. The distances to these hydrogen bonds with R357, S378, and R379 are 3.25 Å, 2.83 Å, and 3.11 Å, respectively. These residues are contained within the binding sites determined by the MOLCAD module in the Sybyl program, therefore compound 21 is considered to bind well to EGR-1.

[0206] also, Figure 5 This is a three-dimensional structural image of the compound 21 complex with EGR-1, predicted using the PyMOL program. (Refer to...) Figure 5 Compound 21 was confirmed to be white and well bound within EGR-1, which consists of green amino acids. The hydrogen bond distances with S378 and R379 were 2.83 Å and 3.11 Å, respectively.

[0207] The same docking experiment as described above was performed using compound 23. Nine compound 23 complexes bound to EGR-1 were obtained. Since the binding energies of these nine complexes ranged from -6.7 kcal / mol to -5.8 kcal / mol, they were considered to be stably bound. Among the nine complexes, the one with the lowest binding energy exhibited a stable conformation of compound 23 bound to EGR-1, and was therefore taken as the product of the binding of compound 23 to EGR-1.

[0208] The interaction between compound 23 and EGR-1 residues was analyzed using the LigPlot program.

[0209] Figure 6 This illustrates the interaction between compound 23 and the amino acid residues of EGR-1. (See reference) Figure 6 Compound 23 forms hydrophobic interactions with eight residues: H358, R379, F377, H382, R375, K366, I361, and R360, and forms a total of four hydrogen bonds with residues R357 and S378 at two sites, respectively. The hydrogen bond distances with R357 are 2.90 Å and 3.95 Å, respectively, while the hydrogen bond distances with S378 are 2.85 Å and 2.96 Å, respectively. These residues are included in the binding sites determined using the MOLCAD module included in the Sybyl program described above, therefore compound 23 is considered to bind well to EGR-1.

[0210] also, Figure 7 The three-dimensional structure of the complex of compound 23 and EGR-1 was predicted using the PyMOL program. See also Figure 7 Compound 23 was confirmed to be well bound to EGR-1, which is composed of green amino acids.

[0211] The same docking experiment as described above was performed using compound 25. Nine compound 25 complexes bound to EGR-1 were obtained. Since the binding energies of these nine complexes ranged from -6.4 kcal / mol to -5.3 kcal / mol, they were considered to be stably bound. Among the nine complexes, the one with the lowest binding energy exhibited a stable conformation of compound 25 bound to EGR-1, and was therefore taken as the product of the binding of compound 25 to EGR-1.

[0212] The interaction between compound 25 and EGR-1 residues was analyzed using the LigPlot program.

[0213] Figure 8 This illustrates the interaction between compound 25 and the amino acid residues of EGR-1. (See reference) Figure 8Compound 25 forms hydrophobic interactions with eight residues: F377, K366, R407, H382, I361, R379, R357, and H358, and forms a hydrogen bond with residue S378. The distance to the hydrogen bond with S378 is 3.06 Å. These residues are included in the binding sites determined using the MOLCAD module included in the Sybyl program described above, therefore compound 25 is considered to bind well in EGR-1.

[0214] also, Figure 9 This is a three-dimensional structural image of the complex of compound 25 and EGR-1 predicted using the PyMOL program. See also... Figure 9 The results confirmed that compound 25 was well bound to EGR-1, which is composed of green amino acids. The formation of S378 and hydrogen bonds is represented in the three-dimensional image as 3.06 Å.

[0215] Experimental Example 4. In vitro ADME analysis of the compounds of the present invention In this experiment, absorption, distribution, metabolism, and excretion (ADME) were predicted on a computer. The method provided by SwissADME (Sci. Rep. 7, 42717 (2017)) was used.

[0216] Compound 21 exhibits moderate solubility in water, good gastrointestinal absorption, and inhibitory activity against cytochrome P450 enzymes CYP1A2, CYP2C9, and CYP3A4. Therefore, compound 21 may interact with other drugs using these enzymes as substrates. However, compound 21 does not inhibit CYP2C19 and CYP2D6, so it is unlikely to interact with other drugs using these enzymes as substrates. Although the proportion of sp3 carbon atoms in compound 21 is at least 0.25 of the total carbon atoms, causing the saturation value to deviate from the baseline and reducing bioavailability, other factors such as molecular size, polarity, solubility, flexibility, and lipophilicity meet the baseline. Therefore, compound 21 is considered to have good bioavailability.

[0217] Compound 23 is well soluble in water, poorly absorbed from the gastrointestinal tract, and inhibits cytochrome P450 enzymes CYP1A2, CYP2C9, and CYP3A4. Therefore, compound 23 may interact with other drugs using these enzymes as substrates. However, compound 23 does not inhibit CYP2C19 or CYP2D6, so it is unlikely to interact with other drugs using these enzymes as substrates. Although the saturation and molecular polarity values ​​deviate from the standards, other parameters of compound 23 (e.g., molecular size, solubility, flexibility, lipophilicity) are within the standards. Therefore, the bioavailability of compound 23 is considered good.

[0218] Compound 25 is well soluble in water and well absorbed from the gastrointestinal tract. Because it inhibits cytochrome P450 enzymes CYP1A2, CYP2C9, and CYP3A4, it may interact with other drugs using these enzymes as substrates. However, since it does not inhibit CYP2C19 and CYP2D6, it is deemed unlikely to interact with other drugs using them as substrates. Compound 25 meets all standard values ​​except for saturation, therefore its bioavailability is considered good.

[0219] Experimental Example 5. Computer-based toxicity analysis of the compounds of this invention. This experiment aimed to predict toxicity on a computer. Toxicity was assessed using the methods described in Pro-Tox-II [Nucleic Acids Res 46 (W1), W257 (2018)].

[0220] Compound 21 has an LD50 of 2100 mg / kg. It exhibits toxicity in all aspects except for hepatotoxicity, carcinogenicity, aryl hydrocarbon receptor, and mitochondrial membrane potential pathway.

[0221] Compound 23 has an LD50 of 2100 mg / kg and exhibits toxicity in all aspects except for immunotoxicity, mutagenicity, aryl hydrocarbon receptor and mitochondrial membrane potential pathway.

[0222] The LD50 of compound 23 is 2100 mg / kg. Apart from hepatotoxicity, carcinogenicity, mutagenicity, and mitochondrial membrane potential pathway, it has no other toxicity.

[0223] Experimental Example 6. Off-target effect of the compounds of the present invention In this experiment, in addition to inhibiting the DNA-binding ability of EGR-1 in computer simulations, we also attempted to predict the target protein to avoid targeting other proteins. The target protein was measured using the methods provided in SwissTarget [Nucleic Acids Research 47 (W1), W357 (2019)].

[0224] Among the target proteins predicted by the non-specific target of compound 21, the highest probability was 0.109, therefore it is considered that there is no non-specific target effect.

[0225] Among the target proteins predicted by the non-specific target of compound 23, the highest probability was 0.109, therefore it is considered that there is no non-specific target effect.

[0226] Among the target proteins predicted by the non-specific target of compound 25, the highest probability was 0.097, therefore it is considered that there is no non-specific target effect.

[0227] Experimental Example 7. Evaluation of the inhibitory effect of the compounds of the present invention on EGR-1 binding using EMSA technology. When nucleic acids bind to proteins, the complexes move more slowly than nucleic acids alone. Therefore, in the Electrophoretic Mobility Shift Assay (EMSA), the nucleic acid band bound to the protein appears higher than the nucleic acid band alone. Using this experimental principle, the compounds (compounds 21, 23, and 25) that showed the highest EGR-1 DNA binding inhibition rates in Experiment 1 were selected. The inhibitory effect of the compounds of this invention on EGR-1 binding was evaluated by analyzing changes in electrophoretic mobility.

[0228] Specifically, protein-DNA binding was analyzed using a LightShift chemiluminescent EMSA kit (ThermoFisherScientific, Waltham, MA, USA). The EGR-1 probe sequence was a biotin-tagged deoxyribonucleotide containing the EGR-1 binding sequence (5'-biotin-AGA GTG TGT CTC CTT CGC ACA CAT C-3'), synthesized by Macrogen (Seoul, South Korea). The EGR-1 protein used in the experiments was obtained by overexpressing human EGR-1 protein in Sf21 insect cells using the method described in Yeo et al.'s paper [Journal of Investigative Dermatology 2021; 141:1851-1855]. Wild-type Sf21 cell extracts were used as negative controls. EGR-1 overexpressing Sf21 cell extract (3 μg), 1 μl of the compound of this invention at a concentration of 20 mM, 50 fmol of biotin-labeled EGR-1-binding deoxy oligonucleotide probe, and 1 μg of poly(dI-dC) (Amersham Pharmacia Biotech Inc) were mixed and reacted for 20 minutes, followed by electrophoresis on a 6% non-denaturing polyacrylamide gel. The electrophoretically separated sample was transferred to a nylon membrane (Immobilon-Ny+ Transfer membrane, Millipore) and immersed in a 5% skim milk solution for 30 minutes. Subsequently, streptavidin conjugated with horseradish peroxidase was added, and after reacting for 15 minutes, unbound proteins were washed away, and the EGR-1-DNA complex was detected on X-ray film using an enhanced chemiluminescence (ECL) detection system (ThermoFisherScientific). The intensity of the EGR-1-DNA complex band was analyzed using ImageJ (National Institute of Health, USA), with the intensity of the sample without added compounds set as 100%, and relative intensities were measured. The experimental results are shown below. Figure 13 .

[0229] Figure 13 The results of electrophoretic mobility shift analysis (EMSA) of the compounds (compounds 21, 23, and 25) according to the invention after application to keratinocytes are shown. Figure 13 As shown, the three selected compounds (compounds 21, 23 and 25) all inhibited EGR-1 DNA binding in a concentration-dependent manner.

[0230] Experimental Example 8. Inhibition Effect of EGR-1 Target TSLP Gene mRNA Expression To confirm whether the compounds of the present invention (compounds 21, 23 and 25) inhibit the expression of EGR-1 target genes TSLP, IL-1β and IL-6, which play a key role in the exacerbation of atopic dermatitis, the inhibitory effect of the compounds on the mRNA expression of the EGR-1 target gene TSLP was evaluated in HaCaT keratinocytes using reverse transcription-polymerase chain reaction (RT-PCR).

[0231] Specifically, HaCaT keratinocytes (purchased from Cell Lines Service (Eppelheim, Germany)) were used at a concentration of 1×10⁻⁶. 6 Inoculation density was achieved by inoculating the culture medium into Dulbecco modified Eagle medium (Invitrogen Life Technologies) containing 10% fetal bovine serum (Invitrogen Life Technologies) and an antimicrobial and antifungal mixture (Invitrogen Life Technologies), and incubating at 37°C in a 5% CO2 incubator, with subculturing every 2 days.

[0232] HaCaT cells were treated with compounds of the present invention (compounds 21, 23, and 25) at concentrations of 0, 5, 10, and 20 μM, followed by treatment with the inflammatory cytokine TNFα at a concentration of 10 ng / ml after 30 minutes, and the cells were harvested after 12 hours. To measure the amount of gene mRNA, total RNA was extracted using TRIzol RNA Isolation Reagents (TRIzol Life Technologies Korea). 0.5 μg of total RNA was reverse transcribed into complementary DNA (cDNA) using the iScript cDNA synthesis kit (Bio-Rad, Hercules, CA, USA) according to the manufacturer's recommended method. PCR was performed using 0.0125 μg of the double-stranded cDNA. Primer bases used for gene amplification were synthesized by Macrogen (Seoul, Korea), and the primer sequences are listed in Table 3 below.

[0233] (Table 3)

[0234] As a control, the housekeeping gene GAPDH (glycerol-3-phosphate dehydrogenase) was used. Double-stranded cDNA was denatured at 95°C for 5 minutes, followed by 30 cycles at 94°C, 65°C, and 70°C (1 minute at 94°C, 2 minutes at 65°C, and 1 minute at 70°C). PCR products were confirmed by 1% agarose gel electrophoresis and ethidium bromide (EtBr) staining. The results are shown below. Figure 14 middle.

[0235] Figure 14 The results of reverse transcription polymerase chain reaction (RT-PCR) following application of the three compounds of the present invention (compounds 21, 23, and 25) to keratinocytes are shown. Figure 14 As shown, the compounds of the present invention (compounds 21, 23, 25) reduce the mRNA expression of EGR-1 target genes TSLP, IL-1β and IL-6 stimulated by TNFα in a dose-dependent manner.

[0236] Experimental Example 9. Therapeutic Effect of DNCB-Induced Atopic Dermatitis in Animal Models (1) Creating an animal model of atopic dermatitis Six- to eight-week-old male BALB / c mice were acclimatized for one week in an animal laboratory at a temperature of 20±2℃, humidity of 50±10%, and a photoperiod of 12 hours. To increase the sensitivity of the ear skin in mice, 4% sodium dodecyl sulfate (SDS) was applied to the ears of mice in the atopic dermatitis control group. Subsequently, 1% 2,4-dinitrochlorobenzene (DNCB; Sigma Chemical Co., St. Louis, MO, USA) was prepared using a mixture of acetone and olive oil (3:1, v / v) and applied to the ears of mice once daily for three consecutive days. After a four-day rest period, 100 μL each of 4% SDS and 0.5% DNCB were applied to the same site five times a week for two consecutive weeks to induce atopic dermatitis. Approximately 2-3 hours later, the following were applied to the same site: (i) the drug solvent (positive control group for atopic dermatitis), (ii) 0.1% tofacitinib (a JAK kinase inhibitor; positive control group), (iii) 0.1% compound 21, (iv) 0.1% compound 23, and (v) 0.1% compound 25, and symptom relief was observed (see [link to relevant documentation]). Figure 15 A).

[0237] (2) The therapeutic effect of the skin application of the compound of the present invention on atopic dermatitis (2-1) Visual observation of the improvement effect on skin inflammation On the last day of the experiment (day 22), all experimental mice were sacrificed, and their skin clinical symptoms were visually observed. Mice treated with compounds 21, 23, and 25 showed significantly improved skin conditions compared to mice treated with only DNCB, with effects similar to the positive control drug tofacitinib (see [link to study]). Figure 15 B).

[0238] (2-2) Improvement effect of skin pathology and histology It has been reported that in atopic dermatitis, repeated inflammatory responses lead to increased proliferation of HaCaT keratinocytes, resulting in an imbalance in differentiation into the stratum corneum and thus thickening of the skin. Numerous major immune cells infiltrate the tissue to promote the inflammatory response. Therefore, the pathological and histological therapeutic effects of the compounds of the present invention (compounds 21, 23, and 25) were analyzed.

[0239] (a) H&E staining Tissue was excised from the ear skin of experimental mice and fixed in 100% acetone solution for 24 hours to prepare paraffin-embedded blocks. The paraffin blocks were sectioned at a thickness of 5 μm and stained with hematoxylin / eosin using the H&E Staining Kit (Abcam, Cambridge, UK). The results showed that, compared with the normal control group, the experimental group with DNCB-induced atopic dermatitis exhibited significantly increased thickness of both the epidermis and dermis (see [link to relevant documentation]). Figure 15 C).

[0240] When quantitatively measuring changes in skin thickness using stained skin tissue, the epidermal thickness of normal skin tissue was 24.47 ± 2.353 μm and the dermal thickness was 143.2 ± 19.61 μm. In tissues treated with DNCB alone, the epidermal thickness was 37.27 ± 4.393 μm and the dermal thickness was 197.5 ± 17.83 μm. In mice treated with the control drug tofacitinib, the epidermal thickness was 27.09 ± 1.719 μm and the dermal thickness was 154.5 ± 11.32 μm. Skin tissue treated with the compounds of the present invention (compounds 21, 23, and 25) exhibited epidermal thicknesses of 25.22 ± 2.993, 22.92 ± 3.617, and 23.46 ± 1.530 μm and dermal thicknesses of 163.4 ± 7.905, 147.5 ± 8.302, and 157.8 ± 17.05 μm (see [link to original text]). Figure 15 D). Based on the above experimental results, it can be confirmed that the compounds of the present invention (compounds 21, 23 and 25) similarly alleviated skin inflammation and inhibited the proliferation of skin keratinocytes as the positive control tofacitinib.

[0241] (b) Inhibition of inflammatory cell infiltration (mast cell infiltration) During an inflammatory response, various inflammatory mediators are secreted when inflammatory cells such as mast cells, neutrophils, and macrophages infiltrate the site of inflammation in the skin, thereby exacerbating the inflammation. Therefore, the efficacy of the compounds of the present invention (compounds 21, 23, and 25) in inhibiting inflammatory cell infiltration was evaluated.

[0242] To measure the number of mast cells infiltrating the inflammatory site, the paraffin-embedded tissue prepared in step (a) was stained with 0.5% toluidine blue (TB). The results showed a significant increase in mast cell infiltration in DNCB-induced atopic dermatitis tissue compared to the normal group, and a significant decrease in mast cell infiltration in the experimental groups treated with tofacitinib and the compounds of this invention (compounds 21, 23, and 25) compared to the positive control group (see [link to relevant documentation]). Figure 16 A).

[0243] Quantitatively measure the number of infiltrating mast cells. In normal skin tissue, per 2.5 cm... 2 There were 11 ± 3 mast cells; in DNCB-induced atopic dermatitis inflamed tissue, there were 38 ± 6 mast cells. In contrast, when tofacitinib was applied, there were 14 ± 5 ​​mast cells. In skin tissue treated with the compounds of the present invention (compounds 21, 23, and 25), the number of mast cells was 18 ± 2, 19 ± 5, and 15 ± 5, respectively (see [link to original text]). Figure 16 B).

[0244] Neutrophils and macrophages were analyzed by immunohistochemistry using myeloperoxidase (MPO) antibody and F4 / 80 antibody. Paraffin-embedded tissues prepared as described in (a) were dewaxed with xylene, hydrated with ethanol, and then incubated at 70°C in 1 mM EDTA (pH 8.0) for 20 min, followed by incubation at room temperature for 1 h in blocking solution containing 7% goat serum. Subsequently, F4 / 80 antibody (Cell Signaling Biotechnology, Beverly Hills, USA) as a macrophage marker and MPO antibody (DAKO, Gloucesterrup, Denmark) as a neutrophil marker were added to each tissue section, and incubation was performed overnight at 4°C. After washing the tissue sections with phosphate buffer, a secondary antibody conjugated to horseradish peroxidase (HRPO) was added, and incubation was performed at room temperature for 1 h. After washing the tissue sections again with phosphate buffer, react them with the 3,3-diaminobenzidine tetrahydrochloride substrate for 5 minutes, followed by H&E staining as a control.

[0245] Compared with the normal group, MPO, a neutrophil marker, was observed in DNCB-induced atopic dermatitis tissue (see [link to study]). Figure 16 C) and macrophage marker F4 / 80 (see C) Figure 16 The expression of D) was significantly increased. In the experimental groups where the control drug tofacitinib or the compounds of the present invention (compounds 21, 23, 25) were applied, the expression was reduced to a level similar to that of normal tissues.

[0246] Based on the above experimental results, it is confirmed that the compounds of the present invention (compounds 21, 23, and 25) have a similar effect to the control drug tofacitinib in reducing the infiltration of mast cells, neutrophils, and macrophages at the site of inflammation that cause the exacerbation of atopic dermatitis symptoms.

[0247] (c) TSLP inhibition and fimbriae restoration Thymic interstitial lymphopoietin (TSLP) is a cytokine belonging to the IL-7 family, secreted by various non-immune cells, such as epithelial cells, smooth muscle cells, fibroblasts, and epidermal keratinocytes. TSLP is produced in large quantities in keratinocytes during skin inflammation, promoting T-lymphocyte differentiation and activating various inflammatory cells. It also stimulates skin sensory neurons to induce itching, thereby exacerbating atopic dermatitis.

[0248] Filamentin is a protein in the stratum corneum of skin that binds to keratin to form a skin barrier crucial for skin hydration and produces natural moisturizing factors. When skin inflammation worsens, filamentin production decreases, leading to damage to the skin barrier, which intensifies dryness and itching, making it easier for various microorganisms, dust particles, and allergens to invade, thus exacerbating the symptoms of atopic dermatitis.

[0249] Therefore, the effects of the compounds of this invention (compounds 21, 23, and 25) on the inhibition of TSLP production and the restoration of fimbriae protein production were analyzed. Specifically, after removing paraffin from the tissue sections, they were placed in a blocking solution containing 1% bovine serum albumin and reacted for 1 hour. Subsequently, TSLP antibody (Novus Biologicals, Centennial, USA) and FLG antibody (Santa Cruz Biotechnology, Dallas, USA) were added to each tissue section, and the reaction was carried out overnight at 4°C. After washing the sections with phosphate buffer, they were reacted at 25°C with a secondary antibody conjugated with rhodamine red-X (Jackson Immuno Research Laboratories, West Grove, USA; 1:300 dilution) for 1 hour. For control staining, nuclear DNA was stained with Hoechst 33258 solution, and fluorescent mounting medium (ProLong Gold anti-fading reagent, Invitrogen, Carlsbad, CA, USA) was added to the coverslip. Fluorescence images were analyzed using an EVOS FL fluorescence microscope (Advanced Microscopy Group; Bothell, WA, USA).

[0250] The results showed that TSLP expression was significantly increased in DNCB-induced atopic dermatitis tissues compared to the normal group. However, in the experimental groups treated with the control drug tofacitinib and the compounds of this invention (compounds 21, 23, and 25), TSLP expression was significantly lower than in the positive control group (see [link to experimental group]). Figure 17 A). Furthermore, a significant reduction in filamentin was observed in atopic dermatitis tissues, while in the experimental groups treated with the control drug tofacitinib and the compounds of this invention (compounds 21, 23, and 25), filamentin expression recovered to levels similar to normal tissues (see [link]). Figure 17 B).

[0251] The experimental results above show that the compounds of the present invention (compounds 21, 23, and 25) are similar to the positive control drug tofacitinib in that they reduce the expression of TSLP, which induces symptoms of atopic dermatitis, while restoring the production of fimbriae, which is essential for skin barrier repair.

[0252] The above description of the present invention is merely illustrative, and those skilled in the art will understand that it can be easily converted into other specific forms without changing the technical concept or essential features of the present invention. Therefore, the above embodiments should be understood in all respects as exemplary and not restrictive.

Claims

1. A pharmaceutical composition for the prevention, improvement, or treatment of atopic dermatitis, wherein, The pharmaceutical composition comprises a compound of formula 1, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient: [Formula 1] In the formula, X represents hydrogen, C1-C6 alkyl, or -CH2-C. 6-10 aryl or And the C1-C6 alkyl group and -CH2-C 6-10 Each aryl group is independently unsubstituted or substituted by one to three halogens; R 1 It is hydrogen or halogen; R 2 It is hydrogen, halogen, hydroxyl, C1-C6 alkyl or C1-C6 alkoxy, and the C1-C6 alkyl and C1-C6 alkoxy are each independently unsubstituted or substituted by 1 to 3 halogens; R 3 R 4 and R 5 Each is independently hydrogen, halogen, hydroxyl or C1-C6 alkoxy, and the C1-C6 alkoxy group is either unsubstituted or substituted by 1 to 3 halogens; R 6 It is hydrogen, halogen, hydroxyl, -NO2, C1-C6 alkyl or C1-C6 alkoxy, and the C1-C6 alkyl and C1-C6 alkoxy are each independently unsubstituted or substituted by 1 to 3 halogens; R 7 It is hydrogen, halogen, hydroxyl, -NO2, -CN, C1-C6 alkyl or C1-C6 alkoxy, and the C1-C6 alkyl and C1-C6 alkoxy are each independently unsubstituted or substituted by 1 to 3 halogens.

2. The pharmaceutical composition according to claim 1, wherein, The compound includes one selected from the following compounds, their isomers, or pharmaceutically acceptable salts thereof: N-(3-(trifluoromethyl)phenyl)hydrazinothiocarbamate; N-(4-fluorophenyl)hydrazinothiocarbamate; N-(4-bromophenyl)hydrazinothiocarbamate; N-(4-cyanophenyl)hydrazinothiocarbamate; N-(4-(trifluoromethyl)phenyl)hydrazinothiocarbamate; N-Ethylhydrazinothiocarbamate; N-Isopropylhydrazine thiocarbamate; N-Isobutylhydrazine thiocarbamate; N-(hydrazylaminothioformyl)benzamide; N-Benzylhydrazine thiocarbamate; N-(m-tolyl)hydrazinothiocarbamate; (Z)-2-(5-fluoro-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(5-fluoro-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(5-fluoro-2-oxoindoline-3-ylidene)-N-(4-fluorophenyl)hydrazylthiocarbamic acid; (Z)-N-(4-bromophenyl)-2-(5-fluoro-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-N-(4-cyanophenyl)-2-(5-fluoro-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(5-fluoro-2-oxoindoline-3-ylidene)-N-(4-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid; (Z)-2-(7-fluoro-2-oxoindoline-3-ylidene)-N-(4-fluorophenyl)hydrazylthiocarbamic acid; (Z)-N-(4-bromophenyl)-2-(7-fluoro-2-oxoindoline-3-ylidene)hydrazylthiocarbamate; (Z)-N-(4-cyanophenyl)-2-(7-fluoro-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(7-fluoro-2-oxoindoline-3-ylidene)-N-(4-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazino-1-thiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(2-methoxyphenyl)hydrazino-1-thiocarbamic acid; (Z)-N-(4-cyanophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(4-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid; (Z)-N-ethyl-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazylthiocarbamate; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(m-tolyl)hydrazinothiocarbamic acid; (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-N-(2-methoxyphenyl)-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-(3-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid; (Z)-N-(4-fluorophenyl)-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(5-bromo-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(3-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-(3-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(4-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(5-bromo-2-oxoindoline-3-ylidene)-N-(4-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-N-(2-methoxyphenyl)-2-(2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(7-bromo-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(5-bromo-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(7-bromo-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-N-(3-nitrophenyl)-2-(2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(5-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(7-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-N-(3-methoxyphenyl)-2-(2-oxoindoline-ylidene)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(3-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(5-bromo-2-oxoindoline-3-ylidene)-N-(3-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-N-(4-methoxyphenyl)-2-(2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(4-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(5-bromo-2-oxoindoline-3-ylidene)-N-(4-methoxyphenyl)hydrazylthiocarbamic acid; N-(3,4-dimethoxyphenyl)hydrazinothiocarbamate; (Z)-N-(4-fluorophenyl)-2-(2-oxoindoline-3-ylidene)hydrazylthiocarbamate; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-chloro-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(6-chloro-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(4-chloro-2-oxoindoline-3-ylidene)-N-(2-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(5-fluoro-2-oxoindoline-3-ylidene)-N-(3-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid; (Z)-N-benzyl-2-(5-fluoro-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(5-fluoro-2-oxoindoline-3-ylidene)-N-(m-tolyl)hydrazinothiocarbamic acid; (Z)-2-(7-fluoro-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(3-(trifluoromethyl)phenyl)hydrazine thiocarbamic acid; (Z)-N-(4-fluorophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid; (Z)-N-(4-bromophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid; (Z)-N-benzyl-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid; (Z)-N-(4-bromophenyl)-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-N-(4-cyanophenyl)-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-N-ethyl-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamate; (Z)-N-benzyl-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-(m-tolyl)hydrazinothiocarbamic acid; (Z)-2-(2-oxo-5-(trifluoromethoxy)indoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-N-(3-nitrophenyl)-2-(2-oxo-5-(trifluoromethoxy)indoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-N-(4-fluorophenyl)-2-(2-oxo-5-(trifluoromethoxy)indoline-3-ylidene)hydrazylthiocarbamate; and (Z)-N-(4-bromophenyl)-2-(2-oxo-5-(trifluoromethoxy)indoline-3-ylidene)hydrazylthiocarbamate.

3. The pharmaceutical composition according to claim 1 or 2, wherein, The pharmaceutical composition can be administered orally, parenterally, or transdermally.

4. A nonprescription pharmaceutical composition for the prevention or improvement of atopic dermatitis, wherein, The over-the-counter pharmaceutical composition comprises, as an active ingredient, a compound of formula 1, an isomer thereof, or a pharmaceutically acceptable salt thereof: [Formula 1] In the formula, X represents hydrogen, C1-C6 alkyl, or -CH2-C. 6-10 aryl or And the C1-C6 alkyl group and -CH2-C 6-10 Each aryl group is independently unsubstituted or substituted by one to three halogens; R 1 It is hydrogen or halogen; R 2 It is hydrogen, halogen, hydroxyl, C1-C6 alkyl or C1-C6 alkoxy, and the C1-C6 alkyl and C1-C6 alkoxy are each independently unsubstituted or substituted by 1 to 3 halogens; R 3 R 4 and R 5 Each is independently hydrogen, halogen, hydroxyl or C1-C6 alkoxy, and the C1-C6 alkoxy group is either unsubstituted or substituted by 1 to 3 halogens; R 6 It is hydrogen, halogen, hydroxyl, -NO2, C1-C6 alkyl or C1-C6 alkoxy, and the C1-C6 alkyl and C1-C6 alkoxy are each independently unsubstituted or substituted by 1 to 3 halogens; R 7 It is hydrogen, halogen, hydroxyl, -NO2, -CN, C1-C6 alkyl or C1-C6 alkoxy, and the C1-C6 alkyl and C1-C6 alkoxy are each independently unsubstituted or substituted by 1 to 3 halogens.

5. A method for preventing or treating atopic dermatitis, comprising administering to a subject a compound of formula 1, an isomer thereof, or a pharmaceutically acceptable salt thereof: [Formula 1] In the formula, X represents hydrogen, C1-C6 alkyl, or -CH2-C. 6-10 aryl or And the C1-C6 alkyl group and -CH2-C 6-10 Each aryl group is independently unsubstituted or substituted by one to three halogens; R 1 It is hydrogen or halogen; R 2 It is hydrogen, halogen, hydroxyl, C1-C6 alkyl or C1-C6 alkoxy, and the C1-C6 alkyl and C1-C6 alkoxy are each independently unsubstituted or substituted by 1 to 3 halogens; R 3 R 4 and R 5 Each is independently hydrogen, halogen, hydroxyl or C1-C6 alkoxy, and the C1-C6 alkoxy group is either unsubstituted or substituted by 1 to 3 halogens; R 6 It is hydrogen, halogen, hydroxyl, -NO2, C1-C6 alkyl or C1-C6 alkoxy, and the C1-C6 alkyl and C1-C6 alkoxy are each independently unsubstituted or substituted by 1 to 3 halogens; R 7 It is hydrogen, halogen, hydroxyl, -NO2, -CN, C1-C6 alkyl or C1-C6 alkoxy, and the C1-C6 alkyl and C1-C6 alkoxy are each independently unsubstituted or substituted by 1 to 3 halogens.

6. A compound selected from any of the following groups, its isomers, or a pharmaceutically acceptable salt thereof: (Z)-N-(4-cyanophenyl)-2-(7-fluoro-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(7-fluoro-2-oxoindoline-3-ylidene)-N-(4-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazino-1-thiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(2-methoxyphenyl)hydrazino-1-thiocarbamic acid; (Z)-N-(4-cyanophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(4-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid; (Z)-N-ethyl-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazylthiocarbamate; (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(7-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(3-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(4-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-chloro-2-oxoindoline-3-ylidene)-N-(2-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(7-fluoro-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(3-(trifluoromethyl)phenyl)hydrazine thiocarbamic acid; (Z)-N-(4-fluorophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid; (Z)-N-(4-bromophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid; (Z)-N-benzyl-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid; (Z)-N-(4-cyanophenyl)-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; and (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-(m-tolyl)hydrazinothiocarbamate.

7. A composition comprising any compound selected from the group consisting of, isomers thereof, or pharmaceutically acceptable salts thereof: (Z)-N-(4-cyanophenyl)-2-(7-fluoro-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(7-fluoro-2-oxoindoline-3-ylidene)-N-(4-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazino-1-thiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(2-methoxyphenyl)hydrazino-1-thiocarbamic acid; (Z)-N-(4-cyanophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(4-(trifluoromethyl)phenyl)hydrazylthiocarbamic acid; (Z)-N-ethyl-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazylthiocarbamate; (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-phenylhydrazine thiocarbamic acid; (Z)-2-(7-bromo-2-oxoindoline-3-ylidene)-N-(3-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(3-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(4-bromo-2-oxoindoline-3-ylidene)-N-(4-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(6-bromo-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(4-chloro-2-oxoindoline-3-ylidene)-N-(2-methoxyphenyl)hydrazylthiocarbamic acid; (Z)-2-(7-fluoro-2-oxoindoline-3-ylidene)-N-(4-nitrophenyl)hydrazylthiocarbamic acid; (Z)-2-(6-methoxy-2-oxoindoline-3-ylidene)-N-(3-(trifluoromethyl)phenyl)hydrazine thiocarbamic acid; (Z)-N-(4-fluorophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid; (Z)-N-(4-bromophenyl)-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid; (Z)-N-benzyl-2-(6-methoxy-2-oxoindoline-3-ylidene)hydrazino-1-thiocarbamic acid; (Z)-N-(4-cyanophenyl)-2-(5-methyl-2-oxoindoline-3-ylidene)hydrazylthiocarbamic acid; and (Z)-2-(5-methyl-2-oxoindoline-3-ylidene)-N-(m-tolyl)hydrazinothiocarbamate.

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