Anti-inflammatory compositions comprising benzofuranyl n-acylhydrazone derivatives

Abstract

The present application relates to a pharmaceutical composition for preventing or treating an inflammatory disease, comprising a benzofuranyl N-acylhydrazone compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. The benzofuranyl N-acylhydrazone compound of the present application can prevent and treat various pathological diseases caused by an inflammatory reaction by inhibiting the initial pathway and process of the inflammatory reaction system of an organism by inhibiting the activity of nuclear factor kappa B (NF-κB), which is a main signaling substance in the inflammatory reaction, and inhibiting inflammatory immune cells and inflammatory cytokines.

Description

Technical Field

[0001] This invention relates to anti-inflammatory compositions comprising benzofuranyl N-acylhydrazone derivatives, and more particularly, to compounds having the effect of inhibiting inflammatory response modulation pathways that may be associated with nuclear factor κB (NF-κB). Background Technology

[0002] Inflammation is a local immune response that occurs when cells or tissues are damaged or destroyed by various factors, such as harmful substances or organisms entering from the outside. It aims to minimize the damage and restore the damaged area to its original state. It is a useful defense mechanism that protects the organism and clears the products of tissue damage. Through this defense mechanism, symptoms such as pain, swelling, redness, or fever can occur, leading to functional impairment. The various factors that induce inflammation include physical factors such as trauma, burns, frostbite, and radiation; chemical factors such as acids; and immunological factors such as antibody reactions. In addition, it can also occur due to imbalances in blood vessels or hormones.

[0003] Under normal circumstances, the aforementioned inflammation plays a role in neutralizing or eliminating pathogenic factors within the body through inflammatory responses and restoring normal tissue and function by regenerating damaged tissue. However, if the degree of inflammation exceeds a certain level or becomes chronic, it can lead to disease states such as chronic inflammation. In clinical practice, inflammatory responses can be observed in almost all diseases.

[0004] Nuclear factor κB (NF-κB) is a transcriptional protein that regulates multiple signal transductions related to inflammatory responses, immune function, aging, and tumors. It has been identified in a wide range of organisms, from fruit flies to mammals. To date, more than 60 genes are known to target NF-κB, and studies have shown that it plays a central role in multiple responses. Therefore, by modulating NF-κB activity, the initial pathways and processes of multiple responses can be selectively controlled, potentially serving as an indicator for disease treatment.

[0005] Proper coordination between the activation and inactivation of nuclear factor κB helps maintain the body's normal state. However, if an inflammatory response is initiated due to excessive activation of nuclear factor κB, the production of inflammatory prostaglandins, eicosanoic acid-like substances, and nitric oxide will abnormally increase. This leads to an excessive and prolonged immune response, inducing aging and the onset, proliferation, and metastasis of tumors, and is associated with various pathological states, including arteriosclerosis and tissue transplant rejection. In particular, nuclear factor κB is highly associated with chronic inflammatory diseases.

[0006] Previous studies have reported that nuclear factor κB, as an inflammatory transcription factor, regulates the expression of various inflammatory genes. Nuclear factor λB (NF-λB) activity is highly associated with cytokine storms or cytokine release syndromes and is known as a therapeutic target factor for these diseases.

[0007] "Cytokine storm" or "cytokine release syndrome" is associated with a variety of conditions, including infectious diseases, non-infectious diseases, autoimmune reactions, and drug side effects. Inflammation is part of a complex biological response of the body's tissues to harmful stimuli such as viruses, damaged cells, or irritants. It is a protective response associated with the immune system, blood vessels, and many proteins. The purpose of inflammation is to remove the initial cause of cell damage and clear dead or dying cells to begin tissue repair. Under normal circumstances, inflammation has a short lifespan, and the recovery of damaged tissue begins on the second or third day after the onset of symptoms. If high levels of inflammatory-inducing proteins (cytokines) are released, the excessive inflammatory response damages blood vessels in the lungs and other tissues, resulting in a cytokine storm or cytokine release syndrome. This inflammatory response induces a decline in the function of various organs in the body.

[0008] Numerous reports indicate that nuclear factor κB (NF-κB) functions as a major regulator of cytokine storms or cytokine release syndromes, and it is particularly considered to play a crucial role in infectious diseases. Infections caused by viruses and other pathogens induce apoptosis and subsequent macrophage activation, followed by the production of reactive oxygen species (ROS), leading to epithelial cell inflammation and cell destruction accompanied by cytokine production. Pro-inflammatory cytokines can also be generated through NF-κB activation that induces transcription of various inflammatory genes, as well as through ROS-dependent inflammatory activation that mediates cytokine production and secretion, resulting in secondary infections. Simultaneously, the disruption of the epithelial cell barrier ultimately leads to increased susceptibility to infection.

[0009] Therefore, recent research on infectious diseases caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has revealed that cytokine storm is a major cause of death in patients who die from viral infection. The nuclear factor κB signaling process has been found to be an important inflammatory signaling process that causes the aforementioned cytokine storm (Trends Endocrinol Metab. 2020 Nov; 31(11):802-803, Pabl o Guisado-Vasco et al.; Front Immunol. 2020 Dec 10; 11:598444., Ralf Kircheis et al.).

[0010] Therefore, there is a need to develop novel compounds that can prevent and treat various pathological diseases caused by inflammatory responses by inhibiting the initial pathways and processes of the organism's inflammatory response system through the regulation of nuclear factor κB, which is the main signaling substance of the inflammatory response.

[0011] Against this backdrop, the present invention confirms that the benzofuranyl N-acylhydrazone series of novel compounds, which have high stability and solubility in vivo, can prevent and treat various pathological diseases caused by inflammatory responses by regulating nuclear factor κB, a major signaling substance in inflammatory responses, and inhibiting the initial pathways and processes of the organism's inflammatory response system, thereby completing the present invention. Summary of the Invention

[0012] The technical problem that the invention aims to solve

[0013] The technical problem to be solved by the present invention is to provide a pharmaceutical composition for the prevention or treatment of inflammatory diseases comprising a benzofuranyl N-acylhydrazone compound, its stereoisomer, or a pharmaceutically acceptable salt thereof.

[0014] Another technical problem to be solved by the present invention is to provide food compositions, feed compositions or health functional foods comprising benzofuranyl N-acylhydrazone compounds, their stereoisomers or pharmaceutically acceptable salts thereof for the prevention or improvement of inflammatory diseases.

[0015] Another technical problem to be solved by the present invention is to provide a cosmetic composition comprising a benzofuranyl N-acylhydrazone compound, its stereoisomer or a pharmaceutically acceptable thereof for relieving skin irritation and / or skin inflammation.

[0016] Technical means to solve the problem

[0017] To address the aforementioned problems, the present invention provides a pharmaceutical composition comprising a compound represented by the following chemical formula 1, its stereoisomer, or a pharmaceutically acceptable salt thereof for the prevention or treatment of inflammatory diseases.

[0018] Chemical Formula 1:

[0019]

[0020] In the above formula, R1 represents H and C. 1-6 Alkyl, C 1-6 alkoxycarbonyl C 1-3 Alkyl or C 1-6 Alkoxy C 1-3 Alkyl group; R2 is a halogen, C 1-6 Alkyl or halogenated C 1-6 Alkyl group; R3 is H, halogen, or C. 1-6 Alkyl, C 1-6 alkoxy or halogenated C1-6 Alkyl group; R4 and R5 are each independently H, halogen, C. 1-6 Alkyl, C 1-6 Alkoxy or C 1-6 Alkyl carbonyl amino.

[0021] The above C 1-6 Alkyl groups may include C 1-3 Alkyl, C 3-6 Alkyl, C 2-4 Alkyl, C 2-6 Alkyl groups, etc.

[0022] Furthermore, the aforementioned C 1-6 Alkoxy groups may include C 1-3 Alkoxy, C 3-6 Alkoxy, C 2-4 Alkoxy, C 2-6 Alkyl groups, etc.

[0023] Furthermore, the aforementioned halogenated C 1-6 Alkyl and Halogenated C 1-6 Alkoxy groups can each have 1 to 10 or 1 to 3 identical or different halogens.

[0024] According to one example, in the above chemical formula 1, R1 is H, -CH3, -CH2CO2CH2CH3, -CH2OCH2CH3, -CH2CH2OCH2CH3, or -CH2CH2OCH3.

[0025] In yet another example, in the above chemical formula 1, R2 is Cl, Br, -CH3, or -CF3.

[0026] According to another example, in the above chemical formula 1, R3 is H, F, Cl, -CH3, -OCH3, or -OCF3.

[0027] According to another example, in the above chemical formula 1, R4 and R5 are each independently H, Cl, -CH3, -OCH3 or -NHCOCH3.

[0028] According to another example, in the above chemical formula 1, R1 is H, -CH3, -CH2CO2CH2CH3, -CH2OCH2CH3, -CH2CH2OCH2CH3 or -CH2CH2OCH3; R2 is Cl, Br, -CH3 or -CF3; R3 is H, F, Cl, -CH3, -OCH3 or -OCF3; and R4 and R5 are each independently H, Cl, -CH3, -OCH3 or -NHCOCH3.

[0029] The compounds of Formula 1 of this invention, their stereoisomers, or pharmaceutically acceptable salts thereof can inhibit the initial pathways and processes of the organism's inflammatory response system and regulate inflammatory immune cells and inflammatory cytokines through nuclear factor κB activity. In particular, they exhibit a dual inhibition effect in regulating both canonical and non-canonical pathways in the modulation of nuclear factor κB. For example, this can be used to demonstrate therapeutic effects on diseases by inhibiting acute inflammatory responses such as severe cytokine release syndrome (CRS) and / or cytokine storms. Especially, they can exhibit excellent therapeutic effects in inflammatory diseases by selectively binding to IKKα, a common major factor in both canonical and non-canonical nuclear factor κB inflammatory signaling processes.

[0030] Therefore, the compounds of Formula 1 of the present invention, their stereoisomers or pharmaceutically acceptable salts thereof can be usefully used in the prevention or treatment of inflammatory diseases.

[0031] In this invention, the aforementioned inflammatory diseases can be selected from dermatitis, cytokine release syndrome, cytokine storm, allergy, nasal polyps, rhinitis, chronic sinusitis, nasal congestion, nasal pruritus, asthma, chronic obstructive pulmonary disease, rheumatoid arthritis, conjunctivitis, keratoconjunctivitis, ophthalmitis, dry eye syndrome, heart failure, arrhythmia, atherosclerosis, multiple sclerosis, inflammatory bowel disease, inflammatory pain, neuropathic pain, osteoarthritis pain, lupus erythematosus, sepsis, Crohn's disease, gout, Sjögren's syndrome, Alzheimer's disease, Parkinson's disease, thyroid autoimmune disease, multiple sclerosis, Guillain-Barré syndrome, autism, hemolytic hemostasis disorder, and thyroid... One or more of the following: goiter, Hashimoto's disease, toxic diffuse goiter, ankylosing spondylitis, polymyalgia rheumatica, celiac disease, ulcerative colitis, type 1 diabetes mellitus, peripheral neuropathy, diabetic peripheral neuropathy, Wegener's granulomatosis, muscle atrophy, fibromyalgia, systemic lupus erythematosus, Behçet's disease, uveitis, glomerulonephritis, pulmonary hemorrhage-nephritis syndrome, autoimmune polyendocrine syndrome, Chag-Strauss syndrome, Henno-Schwarzman syndrome, periarteritis nodosa, arteritis of the high-pressure artery, temporal arteritis, relapsing polychondritis, primary alopecia, severe acute respiratory syndrome, coronavirus type 2 infection, and somnolence.

[0032] According to the present invention, the above-mentioned inflammatory disease can be dermatitis.

[0033] Dermatitis can be one or more of the following: atopic dermatitis, contact dermatitis, allergic dermatitis, acne, eczema, rosacea, seborrheic dermatitis, psoriasis, eczema, pruritus, pruritus, urticaria, idiopathic chronic urticaria, scleroderma, vitiligo, scleroderma, Behcet's disease, and contact infectious impetigo.

[0034] In this invention, cytokine release syndrome and / or cytokine storm are a type of systemic inflammatory response syndrome that can be induced by a variety of factors such as infection or specific drugs.

[0035] More specifically, the cytokine release syndrome and / or cytokine storm of the present invention can be cytokine release syndrome and / or cytokine storm caused by viral infection.

[0036] For example, the aforementioned viruses may be selected from one or more of the following groups: influenza virus, influenza A virus subtype H1N1, avian influenza virus, rhinovirus, adenovirus, coronavirus, Middle East respiratory syndrome coronavirus, severe acute respiratory syndrome coronavirus 2, parainfluenza virus, respiratory syncytial virus, herpesvirus (HSV), human immunodeficiency virus (HIV), and hepatitis virus.

[0037] The compounds of Formula 1 of this invention, their stereoisomers, or pharmaceutically acceptable salts thereof can inhibit rapidly developing cytokine storms by modulating both canonical and atypical pathways in the regulation of nuclear factor κB, thereby demonstrating excellent therapeutic effects. In particular, they show excellent therapeutic and ameliorative effects on acute inflammatory responses, especially viral infections where cytokine storms are a problem, such as Severe Acute Respiratory Syndrome coronavirus 2.

[0038] According to the present invention, the above-mentioned inflammatory disease can be sepsis.

[0039] According to the present invention, the above-mentioned inflammatory disease can be severe acute respiratory syndrome coronavirus type 2 infection.

[0040] In this invention, the benzofuran-N-acylhydrazone compound, its stereoisomer, or a pharmaceutically acceptable salt thereof can inhibit cytokine release syndrome and cytokine storm or exhibit anti-inflammatory activity by inhibiting the activity of nuclear factor κB. In particular, the benzofuran-N-acylhydrazone compound, its stereoisomer, or a pharmaceutically acceptable salt thereof can exhibit dual inhibitory effects in the regulation of nuclear factor κB, regulating both typical and atypical pathways. Furthermore, the signal transduction process of nuclear factor κB can be inhibited by selectively binding to IKKα, an important factor in the inflammatory signal transduction process of nuclear factor κB, and inhibiting the formation of the active structure.

[0041] In this invention, the benzofuranyl N-acylhydrazone compound of this invention, its stereoisomers or pharmaceutically acceptable salts thereof can exhibit anti-inflammatory activity by inhibiting phosphorylation of p65.

[0042] In this invention, the benzofuran-N-acylhydrazone compound, its stereoisomer, or a pharmaceutically acceptable salt thereof exhibits anti-inflammatory activity by reducing the number of inflammatory immune cells.

[0043] In this invention, the benzofuranyl N-acylhydrazone compound, its stereoisomer, or a pharmaceutically acceptable salt thereof can exhibit anti-inflammatory activity by inhibiting the expression of inflammatory cytokines.

[0044] In this invention, the benzofuran-N-acylhydrazone compound, its stereoisomer, or its pharmaceutically acceptable salt exhibits particularly excellent anti-inflammatory effects in dermatitis by inhibiting epidermal hyperplasia caused by inflammatory responses.

[0045] In this invention, the benzofuranyl N-acylhydrazone compound, its stereoisomer, or its pharmaceutically acceptable salt can strongly inhibit inflammatory responses in the lungs while suppressing cytokine release syndrome (CRS) and cytokine storm, thus demonstrating excellent efficacy as a therapeutic agent for sepsis.

[0046] Furthermore, the benzofuran-based N-acylhydrazone compound of the present invention, its stereoisomers, or pharmaceutically acceptable salts thereof can exhibit excellent therapeutic effects in severe acute respiratory syndrome coronavirus 2 infection disease, which is characterized by cytokine storm and severe inflammatory response in the lungs, by inhibiting cytokine release syndrome and cytokine storm.

[0047] Furthermore, the present invention provides food compositions, feed compositions, or health functional foods comprising benzofuranyl N-acylhydrazone compounds represented by the above chemical formula 1, their stereoisomers, or pharmaceutically acceptable salts thereof for the prevention or improvement of inflammatory diseases.

[0048] Furthermore, the present invention provides a cosmetic composition for the prevention or improvement of dermatitis comprising a benzofuranyl N-acylhydrazone compound represented by the above-described chemical formula 1, its stereoisomers, or pharmaceutically acceptable salts thereof.

[0049] Furthermore, the present invention provides a cosmetic composition for relieving skin irritation and / or skin inflammation comprising a benzofuranyl N-acylhydrazone compound represented by the above chemical formula 1, its stereoisomers, or a pharmaceutically acceptable salt thereof.

[0050] Invention Effects

[0051] The benzofuranyl N-acylhydrazone compound of the present invention inhibits the initial pathways and processes of the inflammatory response system of the organism and inhibits inflammatory immune cells and inflammatory cytokines by inhibiting the activity of nuclear factor κB, which is the main signal transducer in the inflammatory response. Therefore, it can be usefully used as a pharmaceutical composition, food composition, feed composition, health functional food or cosmetic composition that can prevent and treat a variety of pathological diseases caused by inflammatory responses. Attached Figure Description

[0052] Figure 1 The results were obtained to confirm the inflammatory response-inhibiting efficacy of the compounds in the embodiments of the present invention through nuclear factor κB reporter gene analysis (assay).

[0053] Figure 2 To confirm the inhibitory effect of the compounds of the embodiments of the present invention on the nuclear factor κB signaling pathway.

[0054] Figure 3 To confirm the results of the compound of the present invention in inhibiting the expression of related factors in the nuclear factor κB pathway.

[0055] Figure 4 The results of the anti-inflammatory effect, confirmed by the naked eye, are shown after treating an embodiment of the present invention with a dermatitis-induced model.

[0056] Figure 5 The results of treating an embodiment of the present invention with a dermatitis-induced model of epidermal hyperplasia were shown by hematoxylin and eosin (H&E) staining, confirming the inhibition of epidermal proliferation.

[0057] Figure 6 The results show a reduction in the number of inflammatory immune cells after treating a dermatitis-inducing model with the compounds of the embodiments of the present invention.

[0058] Figure 7 The results show that the expression of inflammatory cytokines was reduced after treating a dermatitis-induced model with the compounds of the embodiments of the present invention.

[0059] Figure 8 The results show that the compounds in the embodiments of the present invention regulate both typical and atypical pathways.

[0060] Figure 9 The results show that the compounds of the embodiments of the present invention selectively bind to IKKα.

[0061] Figure 10 The results show that the compounds of the embodiments of the present invention inhibit the formation of hexamers of IKKα.

[0062] Figure 11 shows the results confirming the inhibitory effects of the compound of the present invention on mortality from sepsis and the expression of inflammatory cytokines and chemokines in a cytokine storm model (based on a sepsis model). Detailed Implementation

[0063] The present invention will now be described in more detail.

[0064] In this specification, unless otherwise stated, the term "halogen" means F, Cl, Br or I.

[0065] Unless otherwise stated, the term "alkyl" refers to a straight-chain or branched saturated hydrocarbon residue. For example, "C 1-6 "Alkyl" refers to an alkyl group composed of 1 to 6 carbon atoms. Specifically, C 1-6 Alkyl groups may include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, tert-pentyl, sec-pentyl, neopentyl, hexyl, etc.

[0066] Unless otherwise stated, the term "alkoxy" refers to a straight-chain or branched alkyl-oxy residue. For example, "C 1-6 "Alkoxy" refers to alkyl-oxygen groups composed of 1 to 6 carbon atoms. Specifically, C 1-6 Alkoxy groups may include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, n-pentoxy, isopentoxy, tert-pentoxy, secondary-pentoxy, neopentoxy, hexyloxy, etc.

[0067] The terms "haloalkyl" or "haloalkoxy" refer to an alkyl or alkoxy group that is substituted with one or more halogens. Specifically, a haloalkyl or haloalkoxy group can be an alkyl or alkoxy group that is substituted with one or more identical or different halogens.

[0068] The term "substitution" refers to the substitution of a hydrogen atom in a molecule by a substituent, which results in a chemically stable compound without exceeding the specified valence of the atom. For example, "substitution of group A by substituent B" means that the hydrogen atom bonded to the carbon or other atoms forming the skeleton of group A is replaced by substituent B, thus forming a covalent bond between group A and substituent B.

[0069] The present invention provides a pharmaceutical composition comprising a compound represented by the following chemical formula 1, its stereoisomer, or a pharmaceutically acceptable salt thereof for the prevention or treatment of inflammatory diseases.

[0070] Chemical Formula 1:

[0071]

[0072] In the above formula, R1 represents H and C. 1-6 Alkyl, C 1-6 alkoxycarbonyl C 1-3 Alkyl or C 1-6 Alkoxy C 1-3 Alkyl group; R2 is a halogen, C 1-6 Alkyl or halogenated C 1-6 Alkyl group; R3 is H, halogen, or C. 1-6 Alkyl, C 1-6 alkoxy or halogenated C 1-6 Alkyl group; R4 and R5 are each independently H, halogen, C. 1-6 Alkyl, C 1-6 Alkoxy or C 1-6 Alkyl carbonyl amino.

[0073] The above C 1-6 Alkyl groups may include C 1-3 Alkyl, C 3-6 Alkyl, C 2-4 Alkyl, C 2-6 Alkyl groups, etc.

[0074] Furthermore, the aforementioned C 1-6 Alkoxy groups may include C 1-3 Alkoxy, C 3-6 Alkoxy, C 2-4 Alkoxy, C 2-6 Alkyl groups, etc.

[0075] Furthermore, the aforementioned halogenated C 1-6 Alkyl and Halogenated C 1-6 The alkoxy groups may each have 1 to 10 or 1 to 3 identical or different halogens.

[0076] According to one example, in the above chemical formula 1, R1 is H, -CH3, -CH2CO2CH2CH3, -CH2O CH2CH3, -CH2CH2OCH2CH3, or -CH2CH2OCH3.

[0077] In yet another example, in the above chemical formula 1, R2 is Cl, Br, -CH3, or -CF3.

[0078] According to another example, in the above chemical formula 1, R3 is H, F, Cl, -CH3, -OCH3, or -OCF3.

[0079] According to another example, in the above chemical formula 1, R4 and R5 are each independently H, Cl, -CH3, -OCH3 or -NHCOCH3.

[0080] According to another embodiment, in the above chemical formula 1, R1 is H, -CH3, -CH2CO2CH2CH3, -CH2OCH2CH3, -CH2CH2OCH2CH3 or -CH2CH2OCH3; R2 is Cl, Br, -CH3 or -CF3; R3 is H, F, Cl, -CH3, -OCH3 or -OCF3; R4 and R5 are each independently H, Cl, -CH3, -OCH3 or -NHCOCH3.

[0081] Specific examples of compounds of the above chemical formula 1 are as follows.

[0082] 1(Compound 7). (E)-N'-[(2-chloro-1H-indol-3-yl)methylene]-5-methylbenzofuran-2-carbonylhydrazine;

[0083] 2(Compound 13). (E)-N'-[(2-chloro-1-methyl-1H-indol-3-yl)methylene]-5-methylbenzofuran-2-carbonylhydrazine;

[0084] 3(Compound 12). (E)-2-{2-chloro-3-[(2-(5-methylbenzofuran-2-carbonyl)hydrazine)methyl]-1H-indol-1-yl}ethyl acetate;

[0085] 4(Compound 1). (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbazide;

[0086] 5(Compound 14). (E)-N'-{[2-bromo-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbazide;

[0087] 6(Compound 15). (E)-N'-{[1-(2-ethoxyethyl)-2-(trifluoromethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbazide;

[0088] 7(Compound 19). (E)-N'-{[2-chloro-1-(2-methoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbazide;

[0089] 8(Compound 2).(E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbazide;

[0090] (Compound 6). (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-6-methoxy-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbazide;

[0091] 10 (Compound 5). (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-fluoro-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbazide;

[0092] 11(Compound 9).(E)-N'-{[2,5-dichloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbazide;

[0093] 12(Compound 3).(E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-(trifluoromethoxy)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbazide;

[0094] 13(Compound 8). (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-methyl-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbazide;

[0095] 14 (Compound 16). (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methoxybenzofuran-2-carbazide;

[0096] 15 (Compound 17). (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl]methylene}-5-methoxybenzofuran-2-carbazide;

[0097] 16(Compound 20). (E)-5-chloro-N'-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}benzofuran-2-carbazide;

[0098] 17(Compound 21). (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-4,7-dimethylbenzofuran-2-carbazide;

[0099] 18 (Compound 22). (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-4,6-dimethoxybenzofuran-2-carbazide;

[0100] 19 (Compound 18). (E)-N'-{2-[2-((2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl)methylene]hydrazine-1-carbonyl}benzofuran-5-yl)acetamide; and

[0101] 20 (Compound 23). (E)-Ethyl-2-(3-((2-(4,6-dimethoxybenzofuran-2-carbonyl)hydrazine)methyl)-2-methyl-1H-indol-1-yl)acetate.

[0102] 21(Compound 10).(E)-2-(2-methyl-3-((2-(5-methylbenzofuran-2-carbonyl)hydrazine)methyl)-1H-indol-1-yl)ethyl acetate.

[0103] 22(Compound 11).(E)-2-(3-((2-(5-chlorobenzofuran-2-carbonyl)hydrazine)methyl)-2-methyl-1H-indol-1-yl-ethyl acetate.

[0104] 23(Compound 4). (E)-2-(2-chloro-3-((2-(5-methylbenzofuran-2-carbonyl)hydrazine)methyl)-1H-indol-1-yl)methyl acetate.

[0105] The structural formulas of the above compounds (based on compound No.) are shown in the table below.

[0106]

[0107]

[0108] More specifically, the compound represented by chemical formula 1 of the present invention can be a compound represented by chemical formula 2 below.

[0109] Chemical formula 2:

[0110]

[0111] In the above formula, R1 represents H and C. 1-6 Alkyl, C 1-6 alkoxycarbonyl C 1-3 Alkyl or C 1-6 Alkoxy C 1-3 Alkyl group; R2 is a halogen, C 1-6 Alkyl or halogenated C 1-6 Alkyl; R 3a and R 3b Both are H, or when R 3a Or R 3b Any one of them is halogen, C 1-6 Alkyl, C 1-6 alkoxy or halogenated C 1-6 When alkoxy is present, the other is H; R4 is halogen, C. 1-6 Alkyl, C 1-6 Alkoxy or C 1-6 alkyl carbonyl amino; and R5 For H.

[0112] The definitions of R1, R2, R4, and R5 above are the same as those in Chemical Formula 1 above.

[0113] In a specific example, R 3a and R 3b Each can be independently H, F, Cl, -CH3, -OCH3, or -OCF3. More specifically, when R 3a Or R 3b Any one of them is halogen, C 1-6 Alkyl, C 1-6 alkoxy or halogenated C 1-6 When alkoxy is present, the above halogens, C 1-6 Alkyl, C 1-6 alkoxy or halogenated C 1-6 The alkoxy group is F, Cl, -CH3, -OCH3 or -OCF3.

[0114] The present invention comprises pharmaceutically acceptable salts of compounds of the above-described chemical formula 1.

[0115] The aforementioned pharmaceutically acceptable salts should have low toxicity to humans and should not have any adverse effects on the biological activity and physical properties of the parent compound.

[0116] For example, the pharmaceutically acceptable salts mentioned above can be acid addition salts formed from pharmaceutically acceptable free acids.

[0117] The above-mentioned free acids can be inorganic or organic acids. In this case, inorganic acids can be hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, perchloric acid, bromic acid, etc., and 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, pyric acid, aspartic acid, glutamic acid, etc.

[0118] The above-mentioned acid addition salts can be prepared by conventional methods, such as dissolving the compound of the above chemical formula 1 in an excess of acidic aqueous solution, or by precipitating the salts using a water-soluble organic solvent such as methanol, ethanol, acetone or acetonitrile.

[0119] Furthermore, the pharmaceutically acceptable salts mentioned above can be alkali metal salts (sodium salts, etc.) or alkaline earth metal salts (potassium salts, etc.).

[0120] The above-mentioned alkali metal salts or alkaline earth metal salts can be obtained by, for example, dissolving the compound of the above chemical formula 1 in an excess of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering out the undissolved compound salt, and then evaporating and drying the filtrate.

[0121] The pharmaceutically acceptable salts mentioned above can be appropriately modified to be food-grade or cosmetic-grade salts depending on their intended use in food, feed, cosmetics, etc.

[0122] Furthermore, the compounds of the present invention may have a chiral carbon center, thereby allowing R or S to exist as isomers, racemic compounds, single enantiomers or mixtures, single diastereomers or mixtures, and all such stereoisomers and mixtures thereof fall within the scope of the present invention. Stereoisomers can be synthesized with stereospecificity using known, industrially pure starting materials and / or reagents.

[0123] Furthermore, the compounds of the present invention may comprise hydrates and solvates of the compounds of Formula 1 described above. The hydrates and solvates can be prepared by known methods, and are preferably non-toxic and water-soluble. In particular, preferably, the hydrates and solvates may each be hydrates and solvates bound to one to five molecules of water and alcohol solvents (especially ethanol, etc.).

[0124] The compounds of the present invention, namely, the compounds of Formula 1, their stereoisomers, or pharmaceutically acceptable salts thereof, exhibit high in vivo stability and solubility, and possess excellent bioavailability. Such compounds of Formula 1, their stereoisomers, or pharmaceutically acceptable salts thereof can prevent and treat various pathological diseases caused by inflammatory responses by regulating nuclear factor κB, a key signaling molecule in inflammatory responses, and inhibiting the initial pathways and processes of the body's inflammatory response system.

[0125] The benzofuranyl N-acylhydrazone compound represented by the above chemical formula 1, its stereoisomers, or pharmaceutically acceptable salts thereof can be usefully used in the prevention or treatment of inflammatory diseases.

[0126] As used in this application, the term "inflammatory disease" refers to a condition characterized by one or more of the following symptoms: pain (pain caused by the production of harmful substances and nerve stimulation), fever (fever caused by vasodilation), redness (fluidization caused by vasodilation and increased blood flow), swelling (tumor caused by excessive inflow or restricted outflow of fluid), and loss of function (partial or complete, temporary or permanent loss of function).

[0127] Non-limiting examples of inflammatory diseases in which compounds of Formula 1 of the present invention can be used include dermatitis, cytokine release syndrome, cytokine storm, allergy, nasal polyps, rhinitis, chronic sinusitis, nasal congestion, nasal pruritus, asthma, chronic obstructive pulmonary disease, rheumatoid arthritis, conjunctivitis, keratoconjunctivitis, ophthalmitis, xerophthalmia, heart failure, arrhythmia, atherosclerosis, multiple sclerosis, inflammatory bowel disease, inflammatory pain, neuropathic pain, osteoarthritis pain, lupus erythematosus, sepsis, Crohn's disease, gout, Sjögren's syndrome, Alzheimer's disease, Parkinson's disease, thyroid autoimmune disease, multiple sclerosis, Guillain-Barré syndrome, autism, and hemolytic hemostasis. Abnormalities, thyroiditis, Hashimoto's disease, toxic diffuse goiter, ankylosing spondylitis, polymyalgia rheumatica, celiac disease, ulcerative colitis, type 1 diabetes, peripheral neuropathy, diabetic peripheral neuropathy, Wegener's granulomatosis, muscle atrophy, fibromyalgia, systemic lupus erythematosus, Behçet's disease, uveitis, glomerulonephritis, pulmonary hemorrhage-nephritis syndrome, autoimmune polyendocrine syndrome, Chag-Strauss syndrome, Henno-Shull syndrome, periarteritis nodosa, arteritis of the high-pressure artery, temporal arteritis, relapsing polychondritis, primary alopecia, somnolence, severe acute respiratory syndrome, coronavirus type 2 infection, etc., but not limited to these.

[0128] In one embodiment of the present invention, the benzofuranyl N-acylhydrazone compound represented by Formula 1, its stereoisomers, or pharmaceutically acceptable salts thereof can be usefully used in the treatment of cytokine release syndrome and / or cytokine storm.

[0129] Cytokine release syndrome and / or cytokine storm is a systemic inflammatory response syndrome that can be induced by a variety of factors, such as infection or specific drugs.

[0130] Elevated levels of inflammatory cytokines can also be a symptom of the aforementioned syndrome. For example, it is characterized by increases in TNF-α, IFN-γ, IL-1β, IL-2, IL-6, IL-8, IL-10, IL-13, GM-CSF, IL-5, fractalkine, or in combinations thereof or downstream combinations thereof.

[0131] More specifically, the cytokine release syndrome and / or cytokine storm of the present invention can be cytokine release syndrome and / or cytokine storm caused by viral infection.

[0132] For example, the aforementioned viruses may be selected from one or more of the following groups: influenza virus, influenza A virus H1N1 subtype, avian influenza virus, rhinovirus, adenovirus, coronavirus, Middle East respiratory syndrome coronavirus, severe acute respiratory syndrome coronavirus type 2, parainfluenza virus, respiratory syncytial virus, herpesvirus, human acquired immunodeficiency virus, and hepatitis virus.

[0133] The compounds of Formula 1 of this invention, their stereoisomers, or pharmaceutically acceptable salts thereof can inhibit rapidly developing cytokine storms by regulating both canonical and atypical pathways in the regulation of nuclear factor κB, thereby demonstrating excellent therapeutic effects. In particular, they show excellent therapeutic and ameliorative effects on acute inflammatory responses such as Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), especially on viral infections where cytokine storms are a problem.

[0134] In one embodiment of the present invention, the inflammatory disease can be dermatitis. In the present invention, the aforementioned dermatitis includes, but is not limited to, one or more of the following: atopic dermatitis, contact dermatitis, allergic dermatitis, acne, eczema, rosacea, seborrheic dermatitis, psoriasis, pruritus, pruritus dermatitis, urticaria, idiopathic chronic urticaria, scleroderma, vitiligo, scleroderma, Behçet's disease, and impetigo contagious.

[0135] In one embodiment of the invention, dermatitis may be atopic dermatitis. In this invention, the term "atopic dermatitis" refers to an inflammatory and recurrent, non-infectious, pruritic, chronic skin disorder.

[0136] Atopic dermatitis, a specific form of dermatitis named according to its location, appearance, or the stressor that triggers it, is also included in the term "atopic dermatitis" of this invention. Atopic dermatitis presents with symptoms such as dry, eczematous skin, papules, and severe itching. Epidermal hyperplasia, epidermal proliferation, and aggregation of lymphocytes and mast cells have been confirmed in lesion samples from patients with atopic dermatitis. Patients with atopic dermatitis typically suffer from severe itching, which can trigger inflammation of the skin lesions, further exacerbating the itching and thus worsening the clinical symptoms.

[0137] According to the present invention, the above-mentioned inflammatory disease can be sepsis.

[0138] For sepsis exhibiting symptoms of a cytokine storm and / or severe cytokine syndrome with acute inflammatory response, the benzofuranyl N-acylhydrazone compound of the present invention, its stereoisomers, or pharmaceutically acceptable salts thereof inhibit the expression of inflammatory cytokines or chemokines, thereby exhibiting anti-inflammatory effects and thus inhibiting the development of death or disease caused by sepsis.

[0139] According to the present invention, the aforementioned inflammatory disease can be severe acute respiratory syndrome coronavirus 2 infection. The disease caused by severe acute respiratory syndrome coronavirus 2 infection can be an inflammatory disease of the respiratory tract. The aforementioned severe acute respiratory syndrome coronavirus 2 infection can manifest symptoms after an incubation period of, for example, 2 to 14 days after viral infection. These symptoms include, for example, gastrointestinal symptoms such as high fever, cough, wheezing, pneumonia, and diarrhea; organ failure (renal failure, renal dysfunction, etc.); septic shock; and in severe cases, death, encompassing all symptoms arising from the aforementioned viral infection. In particular, most of the fatal symptoms of the aforementioned severe acute respiratory syndrome coronavirus 2 infection are highly associated with cytokine storm and / or symptoms of severe cytokine syndrome. The benzofuranyl N-acylhydrazone compound, stereoisomer, or pharmaceutically acceptable salt thereof represented by the above chemical formula 1 of the present invention can selectively bind to IKKα, an important factor in the inflammatory signal transduction process of nuclear factor κB, to inhibit the formation of active structures. It can inhibit and treat the symptoms of the above-mentioned cytokine storm and / or severe cytokine syndrome by regulating both typical and atypical pathways.

[0140] The benzofuranyl N-acylhydrazone compound represented by the above chemical formula 1 of the present invention, its stereoisomers or pharmaceutically acceptable salts thereof may inhibit the symptoms of cytokine storm and / or cytokine syndrome and exhibit anti-inflammatory activity by inhibiting the activity of nuclear factor κB, but is not limited thereto.

[0141] The benzofuranyl N-acylhydrazone compound of the present invention was confirmed to have an inhibitory effect on the nuclear factor κB inflammatory response pathway, which is the main signal transduction system for regulating inflammatory responses, through cell-based reporter gene analysis. It was confirmed that the compound exhibits anti-inflammatory activity by inhibiting the phosphorylation of p65, which is an important factor in the nuclear factor κB signal transduction process.

[0142] The benzofuranyl N-acylhydrazone compound of the present invention inhibits the formation of active structures by selectively binding to IKKα, which is an important factor in the inflammatory signal transduction process of nuclear factor κB.

[0143] The benzofuranyl N-acylhydrazone compound of the present invention can inhibit and treat the symptoms of cytokine storm and / or severe cytokine syndrome by regulating the dual inhibitory effects of the typical and atypical pathways of nuclear factor κB.

[0144] The dual inhibitory effect mentioned above shows remarkable efficacy in the treatment of related diseases, especially in cases of a rapid increase in inflammatory cytokines that may occur in the lungs or an increase in inflammatory response values ​​caused by sepsis.

[0145] The benzofuranyl N-acylhydrazone compound, stereoisomer, or pharmaceutically acceptable salt thereof represented by the above chemical formula 1 of the present invention can exhibit anti-inflammatory activity by reducing the number of inflammatory immune cells, and can also exhibit anti-inflammatory activity by inhibiting the expression of inflammatory cytokines.

[0146] In particular, it exhibits excellent therapeutic effects in the skin and other areas by inhibiting epidermal proliferation caused by inflammation.

[0147] The pharmaceutical compositions of the present invention may also contain one or more active ingredients that exhibit anti-inflammatory or antiviral activity.

[0148] The compositions of the present invention may further comprise pharmaceutically acceptable additives, in which case the pharmaceutically acceptable additives may include starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silica, dicalcium phosphate, lactose, mannitol, maltose, gum arabic, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, omega-4, sodium starch glycolate, palm wax, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, calcium stearate, white sugar, glucose, sorbitol, and talc, etc. Preferably, the compositions comprise 0.1 to 90 parts by weight of the pharmaceutically acceptable additives of the present invention, but are not limited thereto.

[0149] That is, the compositions of the present invention can be administered orally and parenterally in various dosage forms in actual clinical administration. They can be prepared using commonly used fillers, expanders, binders, wetting agents, disintegrants, surfactants, and other diluents or excipients. Solid dosage forms for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid dosage forms are prepared by mixing at least one excipient with the Tianming extract, such as starch, calcium carbonate, sucrose, lactose, or gelatin. Furthermore, in addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. Liquid dosage forms for oral administration include suspensions, internal solutions, emulsions, and syrups, etc. In addition to water and liquid paraffin as simple diluents, they can also contain various excipients, such as wetting agents, sweeteners, flavoring agents, preservatives, etc. Preparations for parenteral administration can include sterile aqueous solutions, non-aqueous solvents, suspensions, oils, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspension solvents can include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. The base of suppositories can include Witepsol, polyethylene glycol, Tween 61, cocoa butter, glyceryl laurate, and glycerin gelatin.

[0150] The compositions of the present invention can be administered orally or parenterally according to the desired method. Parenteral administration can be performed via transdermal, topical, intraperitoneal, intravenous, subcutaneous, intravenous, intramuscular, or intrathoracic injection. For example, transdermal administration is possible. The dosage range varies depending on the patient's weight, age, sex, health status, diet, administration time, administration method, metabolic rate, and disease severity.

[0151] The compositions of the present invention are administered in pharmaceutically effective amounts. In this invention, a "pharmaceutically effective amount" refers to a sufficient amount to treat a disease in a reasonable benefit / risk ratio suitable for medical treatment. The level of the effective dose can be determined based on factors including patient type, disease severity, drug activity, drug sensitivity, timing of administration, route of administration and metabolic rate, duration of treatment, concurrent medications, and other factors well-known in the medical field. The compositions of the present invention can be administered as a single therapeutic agent or in combination with other therapeutic agents, sequentially or simultaneously with existing therapeutic agents, and as a single or multiple dose. Importantly, after considering all the above factors, the dosage is determined to achieve the maximum effect with the minimum amount without side effects, which can be easily determined by a person skilled in the art.

[0152] Specifically, the marketable dosage of the compounds of the present invention can vary depending on the patient's age, sex, and weight. Typically, the dosage is 0.1 mg to 100 mg per kg of body weight. Preferably, it is 0.5 mg to 10 mg daily, administered every other day or once to five times a week, or divided into one to five daily doses. However, the dosage can be increased or decreased depending on the route of administration, the severity of the disease, sex, weight, and age. The above dosage is not in any way limiting the scope of the present invention.

[0153] The compounds of the present invention or pharmaceutical compositions comprising them can be used as anti-inflammatory agents.

[0154] Therefore, the present invention provides the use of a compound of the above-described chemical formula 1, its stereoisomers or pharmaceutically acceptable salts thereof for the prevention or treatment of inflammatory diseases.

[0155] Furthermore, the present invention provides the use of the compound of the above-described chemical formula 1, its stereoisomers or pharmaceutically acceptable salts thereof in the preparation of a medicament for the prevention or treatment of inflammatory diseases.

[0156] Furthermore, the present invention provides a method for preventing or treating inflammatory diseases, comprising the step of administering a compound of the above-described chemical formula 1, its stereoisomers, or pharmaceutically acceptable salts thereof to a subject in need.

[0157] "Prevention" refers to all behaviors that inhibit or delay the occurrence, spread, and recurrence of the above-mentioned diseases by administering the above-mentioned compounds, while "treatment" refers to all behaviors that improve or beneficially change the above-mentioned diseases by administering the above-mentioned compounds.

[0158] The term "required subjects" refers to all animals, including humans (patients), such as monkeys, cattle, horses, sheep, pigs, chickens, turkeys, quails, cats, dogs, mice, rats, rabbits, guinea pigs, etc., specifically mammals. Furthermore, the term "required subjects" can refer to biological samples.

[0159] Furthermore, "administration" refers to providing a specified substance to a desired object by any appropriate method, and the administration pathway of the compounds of the present invention can be through any conventional pathway capable of reaching the target tissue.

[0160] The benzofuranyl N-acylhydrazone compound of the present invention exhibits excellent anti-inflammatory efficacy by inhibiting the phosphorylation of p65(RelA), an important factor in the nuclear factor κB signaling process, through the inhibition of the nuclear factor κB inflammatory response pathway. Furthermore, it demonstrates anti-inflammatory activity by reducing the number of inflammatory immune cells and inhibiting the expression of inflammatory cytokines.

[0161] The present invention also provides compounds of the above-described chemical formula 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof for the prevention or treatment of inflammatory diseases.

[0162] The present invention also provides the use of the compounds of the above-described chemical formula 1, their stereoisomers or pharmaceutically acceptable salts thereof in the preparation of medicaments for treating inflammatory diseases.

[0163] Therefore, compositions comprising compounds of Formula 1 above, their stereoisomers or pharmaceutically acceptable salts thereof can be usefully used as food compositions for the prevention or improvement of inflammatory diseases.

[0164] In this invention, the above-mentioned food composition includes health functional foods.

[0165] The food composition of the present invention may also include other food compositions, additives commonly found in health functional foods or beverages.

[0166] For example, the food composition of the present invention may contain sweeteners such as white sugar, crystalline fructose, glucose, D-sorbitol, mannitol, isomaltooligosaccharide, steviol glycosides, aspartame, acesulfame potassium, and sucrose; acidulants such as anhydrous citric acid, DL-malic acid, succinic acid and its salts; preservatives such as benzoic acid and its derivatives; various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavoring agents, coloring agents and thickeners (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective pectin thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, and carbonating agents used in carbonated beverages. Furthermore, the food composition of the present invention may contain natural fruit juice and fruit pulp used in the preparation of vegetable beverages. The proportions of the above additives may be used in the range of approximately 20 parts by weight or less per 100 parts by weight of the food composition.

[0167] When the food composition of the present invention is a beverage, it may also contain flavoring agents or natural carbohydrates commonly found in beverages. The aforementioned natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, or sugar alcohols such as xylitol, sorbitol, and erythritol. Furthermore, the aforementioned flavoring agents may be natural flavoring agents such as sematriol or stevia extract (lebdoside A, glycyrrhizic acid, etc.), or synthetic flavoring agents such as saccharin and aspartame. When the aforementioned food composition is a beverage, it typically contains about 1g to 20g of natural carbohydrates per 100mL of composition, preferably about 5g to 12g.

[0168] The food composition of the present invention can be prepared in the form of powder, granules, tablets, capsules or beverages, and can be used as food, beverage, chewing gum, tea, vitamin complex, or health supplement.

[0169] The benzofuranyl N-acylhydrazone compound of the present invention exhibits excellent anti-inflammatory efficacy by inhibiting the phosphorylation of p65 (Re1A), an important factor in the nuclear factor κB signaling process, through the inhibition of the nuclear factor κB inflammatory response pathway. Furthermore, it improves or prevents the pathogenesis of cytokine release syndrome, cytokine storm, etc., by reducing the high expression of inflammatory cytokines.

[0170] Therefore, compositions comprising the benzofuranyl N-acylhydrazone compound represented by Formula 1 of the present invention can be usefully used as feed compositions for the prevention or improvement of inflammatory diseases.

[0171] In this invention, the aforementioned feed can be insect, fish, poultry, or mammal feed. Preferably, it can be feed for livestock or aquatic products that have been domesticated to wild habits and are expected to increase agricultural yields, as defined in Article 2, Paragraph 1 of the Korean Livestock Industry Act and Article 2 of its Enforcement Regulations. The aforementioned livestock can be cattle, horses, mules, donkeys, goats, sheep, deer, pigs, and rabbits; poultry can be chickens, turkeys, ducks, ostriches, geese, quails, etc., preferably chickens. There are no restrictions on any species suitable for raising livestock to produce livestock products. The aforementioned "livestock products" refer to meat, milk, eggs, honey, and their processed products, raw hides (including raw fur), raw wool, and other livestock products produced by livestock as defined in Article 2, Paragraph 3 of the Korean Livestock Industry Act, as stipulated by the Ministry of Agriculture and Forestry regulations. Furthermore, it may also include pets such as dogs and cats, but is not limited to these.

[0172] In this invention, the term "feed" refers to a natural or artificial, prescribed, single-use, or otherwise formulated ingredient intended for consumption, ingestion, digestion, or suitability for animals. As one embodiment, the feed composition of this invention containing mealworm larvae extract may include concentrates, roughage, and / or specialty feeds.

[0173] Concentrated feed can include seeds of cereals such as wheat, oats, and corn; bran, including rice bran, wheat bran, and barley bran, which are byproducts obtained through the refining of grains; oil residue, which is a byproduct obtained after pressing soybeans, rapeseed, sesame, flaxseed, and coconut; residual starch, which is the main component of starch residue, left after starch is extracted from sweet potatoes and potatoes; fish meal; fish residue; fish juice, which is a concentrated form of fresh liquid obtained from fish; meat meal; blood meal; feather meal; skim milk powder; dried whey, which is the residual liquid from the preparation of cheese from milk and casein from skim milk; and other animal feeds such as yeast, chlorella, and seaweed.

[0174] Roughage includes wild grasses, pasture grasses, and uncut grasses; feed-grade turnips, feed-grade beets, and root vegetables such as turnip cabbage (used as a radish); silage, which is stored as feed by lactic acid fermentation of grasses, uncut crops, and grains in silos; hay made by cutting and drying wild grasses and pasture grasses; straw used for breeding livestock; and leaves of legumes. Specialty feeds include mineral feeds such as oyster shells and rock salt; urea feeds such as urea or its derivatives like diurea isobutane; supplements that are easily lacking when formulated solely from natural feed ingredients; and feed additives added in trace amounts to compound feeds to improve their storability.

[0175] The benzofuranyl N-acylhydrazone compound of the present invention exhibits excellent anti-inflammatory efficacy by inhibiting the phosphorylation of p65 (Re1A), an important factor in the nuclear factor κB signaling process, through the inhibition of the nuclear factor κB inflammatory response pathway. Furthermore, it improves or prevents the pathogenesis of cytokine release syndrome, cytokine storm, etc., by reducing the high expression of inflammatory cytokines.

[0176] Therefore, the present invention can provide cosmetic compositions for the prevention or improvement of dermatitis comprising the benzofuranyl N-acylhydrazone compound represented by Chemical Formula 1, its stereoisomers, or pharmaceutically acceptable salts thereof. Furthermore, the present invention can provide cosmetic compositions for relieving skin irritation and / or skin inflammation comprising the benzofuranyl N-acylhydrazone compound represented by Chemical Formula 1, its stereoisomers, or pharmaceutically acceptable salts thereof.

[0177] In this invention's specification, "skin injury" refers to any skin injury that can be improved, or prevented or promoted for recovery by inhibiting, reducing, or lowering the expression levels of cytokines. In this invention's specification, "skin irritation" refers to any irritation that may occur in the skin, such as itching, fever, erythema, or skin edema.

[0178] The cosmetic compositions of the present invention can be prepared into any dosage form commonly prepared in the relevant art, for example, as solutions, suspensions, emulsions, balms, gels, creams, lotions, powders, soaps, surfactant-containing detergents, oils, foundation creams, emulsion foundation creams, wax foundation creams, and sprays, but are not limited thereto. More specifically, they can also be prepared into dosage forms such as softening toners, nourishing toners, nourishing creams, massage creams, serums, eye creams, cleansing creams, cleansing foams, cleansing waters, masks, sprays, or powders.

[0179] When the dosage form of the present invention is an ointment, cream, or gel, animal oil, vegetable oil, wax, paraffin, starch, astragalus gum, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide can be used as carrier components.

[0180] When the dosage form of the present invention is a powder or a spray, the carrier component may be lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder. In particular, in the case of a spray, it may also contain a propellant such as chlorofluorocarbons, propane / butane or dimethyl ether.

[0181] When the dosage form of the present invention is a solution or emulsion, the carrier component may be a solvent, solubilizer or emulsifier, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butanediol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan.

[0182] In the case where the dosage form of the present invention is a suspension, the carrier component can be a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isooctadecyl alcohol, polyoxyethylene sorbitan ester and polyoxyethylene sorbitan anhydride ester, microcrystalline cellulose, aluminum hydroxide, benzoate, agar or astragalus gum, etc.

[0183] In the case where the dosage form of the present invention is a detergent containing a surfactant, the carrier component may be fatty alcohol sulfate, fatty alcohol ether sulfate, sulfosuccinate monoester, hydroxyethanesulfonate, imidazole derivative, methyl taurine, sarcosinate, fatty acid amide ether sulfate, cocamidopropyl betaine, fatty alcohol, glycerol fatty acid ester, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ether, etc.

[0184] The following preferred embodiments are presented to aid in understanding the present invention. However, the embodiments described below are merely illustrative of the present invention. Those skilled in the art should understand that various changes and modifications can be made within the scope and technical concept of the present invention, and these changes and modifications should naturally be included within the scope of the appended claims.

[0185] The abbreviations used in the following embodiments are defined as follows.

[0186] EA: Ethyl acetate; EtOH: Ethanol

[0187] DMF: Dimethylformamide; n-Hex: n-hexane

[0188] PrOH: Propanol DMSO: Dimethyl sulfoxide

[0189] Et: Ethyl

[0190] Preparation of aromatic substituted benzofuran-2-carbonylhydrazine derivatives

[0191]

[0192] Preparation Example 1: Ethyl 5-methylbenzofuran-2-carboxylate

[0193]

[0194] 5-Methylsalicylaldehyde (1.40 g, 10.3 mmol) was dissolved in DMF (30 mL), and Ce₂SO₄ (10.07 g, 30.9 mmol) and ethyl bromoacetate (1.38 mL, 12.4 mmol) were added. The mixture was reacted at 70 °C for 15 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO₄, filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA = 10:1) to obtain 1.28 g of the title compound (61% yield, colorless liquid).

[0195] 1 HNMR (400MHz, CDCl3): δ7.49-7.46 (m, 3H), 7.26 (m, 1H), 4.45 (q, J=7.2Hz, 2H), 2.46 (s, 3H), 1.44 (t, J=7.2Hz, 3H); 13 CNMR (100MHz, CDCl3): δ159.85, 154.37, 145.92, 133.51, 129.27, 127.22, 122.45, 113.74, 112.02, 61.61, 21.45, 14.51; mp 38-39℃

[0196] Preparation Example 2: 5-Methylbenzofuran-2-carbonylhydrazine

[0197]

[0198] Ethyl 5-methylbenzofuran-2-carboxylate (2.04 g, 10.0 mmol) obtained in Preparation Example 1 was dissolved in EtOH (30 mL), and hydrazine monohydrate (1.50 g, 30.0 mmol) was added and the mixture was refluxed for 24 hours. The solid obtained by vacuum distillation of the reaction solution was washed with water and dried to obtain 1.72 g of the title compound (90% yield, white solid).

[0199] 1 HNMR (400MHz, DMSO-d6): δ9.97 (s, 1H), 7.52-7.49 (m, 2H), 7.43 (s, 1H), 7.25 (dd, J=8.4, 1.6Hz, 1H), 4.55 (brs, 2H), 2.40 (s, 3H); 13CNMR (100MHz, DMSO-d6): δ157.92, 152.64, 148.48, 132.68, 127.84, 127.11, 122.04, 111.27, 108.47, 20.81; MS (MALDI-TOF): m / z 213[M+Na] + ;mp 159℃

[0200] Preparation Example 3: Ethyl 5-methoxybenzofuran-2-carboxylate

[0201]

[0202] 2-Hydroxy-5-methoxybenzaldehyde (1.25 mL, 10.0 mmol) was dissolved in DMF (30 mL), followed by the addition of K₂CO₃ (6.91 g, 50.0 mmol) and ethyl bromoacetate (1.33 mL, 12.0 mmol), and the reaction was carried out at 70 °C for 15 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO₄, filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA = 3:1) to give 1.1 g of the title compound (50% yield, white solid).

[0203] 1 HNMR (400MHz, CDCl3): δ7.50-7.47 (m, 2H), 7.08-7.05 (m, 2H), 4.45 (q, 2H, J=7.2Hz), 3.86 (s, 3H), 1.44 (t, 3H, J=7.2Hz); mp 53℃

[0204] Preparation Example 4: 5-Methoxybenzofuran-2-carbonylhydrazine

[0205]

[0206] Ethyl 5-methoxybenzofuran-2-carboxylate (1.03 g, 4.68 mmol) obtained in Preparation Example 3 above was dissolved in EtOH (30 mL), hydrazine monohydrate (702.8 mg, 14.04 mmol) was added, and the solution was refluxed for 24 hours. After washing with CH2Cl2, the solid obtained by vacuum distillation of the reaction solution was dried to give 918 mg of the title compound (95% yield, white solid).

[0207] 1HNMR (400MHz, DMSO-d6): δ9.98 (s, 1H), 7.53 (d, 1H, J=9.0Hz), 7.44 (s, 1H), 7. 25 (d, 1H, J=2.7Hz), 7.02 (dd, 1H, J=9.0, 2.7Hz), 4.55 (brs, 2H), 3.79 (s, 3H); 13 CNMR (100MHz, DMSO-d6): δ157.85, 155.97, 149.10, 149.03, 127.65, 115.80, 112.34, 108.34, 104.09, 55.58; MS (MALDI-TOF): m / z 229.0[M+Na] + 245[M+K] + ;mp 163-164℃

[0208] Preparation Example 5: 5-Chlorobenzofuran-2-carbonylhydrazine

[0209]

[0210] At 0 °C, oxalyl chloride (257.2 μL, 3.0 mmol) and DMF (50 μL) were added to CH2Cl2 (5 mL) and stirred for 5 min. Then, a CH2Cl2 / DMF mixture (5 mL, 4:1) of 5-chlorobenzofuran-2-carboxylic acid (393.2 mg, 2.0 mmol) was added and stirred at room temperature for 2 h. The reaction solution was distilled under reduced pressure, and CH2Cl2 (5 mL) was added again. Then, a CH2Cl2 (5 mL) solution of hydrazine monohydrate (120.1 mg, 2.4 mmol) and N,N-diisopropylethylamine (DIEA, 1.74 mL, 10 mmol) were added at 0 °C and stirred at room temperature for 15 h. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over Na2SO4, filtered, and distilled under reduced pressure. The residue was washed with CH2Cl2 and dried to obtain 295 mg of the title compound (70% yield, pale yellow solid).

[0211] 1 HNMR (400MHz, DMSO-d6): δ10.10 (s, 1H), 7.86 (d, J=2.4Hz, 1H), 7.68 (d, J=9.2Hz, 1H), 7.48 (s, 1H), 7.46 (dd, J=9.2, 2.4Hz, 1H), 4.59 (brs, 2H); 13CNMR (100MHz, DMSO-d6): δ157.41, 152.66, 149.79, 128.65, 127.99, 126.56, 122.02, 113.44, 108.25; MS (MALDI-TOF): m / z 233[M+Na] + ;mp 174-175℃

[0212] Preparation Example 6: Ethyl 4,7-dimethylbenzofuran-2-carboxylate

[0213]

[0214] 3,6-Dimethylsalicylaldehyde (450.5 mg, 3.0 mmol) was dissolved in DMF (30 mL), and Ce₂SO₄ (2.93 g, 9.0 mmol) and ethyl bromoacetate (399.2 μL, 3.60 mmol) were added. The mixture was reacted at 70 °C for 15 h. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO₄, filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA = 20:1) to obtain 360 mg of the title compound (55% yield, colorless liquid).

[0215] 1 HNMR (400MHz, CDCl3): δ7.56 (s, 1H), 7.14 (d, J=7.2Hz, 1H), 6.99 (d, J=7.2Hz , 1H), 4.45 (q, J=7.2Hz, 2H), 2.55 (s, 3H), 2.52 (s, 3H), 1.44 (t, J=7.2Hz, 3H); 13 CNMR (100MHz, CDCl3): δ159.91, 154.90, 145.08, 130.19, 128.30, 126.63, 123.90, 119.85, 113.01, 61.43, 18.29, 15.00, 14.46

[0216] Preparation Example 7: 4,7-Dimethylbenzofuran-2-carbonylhydrazine

[0217]

[0218] Ethyl 4,7-dimethylbenzofuran-2-carboxylate (290.3 mg, 1.33 mmol) obtained in Preparation Example 6 was dissolved in EtOH (20 mL), and hydrazine monohydrate (199.7 mg, 3.99 mmol) was added and the mixture was refluxed for 24 hours. The solid obtained by vacuum distillation of the reaction solution was washed with water and dried to obtain 236 mg of the title compound (yield 87%, white solid).

[0219] 1 HNMR (400MHz, DMSO-d6): δ9.97 (s, 1H), 7.55 (s, 1H), 7.13 (d, J=7.4Hz, 1H), 7.00 (d, J=7.4Hz, 1H), 4.59 (brs, 2H), 2.46 (s, 6H, CH3x2); 13 CNMR (100MHz, DMSO-d6): δ158.05, 153.07, 147.65, 129.34, 127.07, 126.47, 123.64, 118.65, 107.94, 17.86, 14.38; MS (MALDI-TOF): m / z 227[M+Na] + ;mp 187℃

[0220] Preparation Example 8: Ethyl 4,6-dimethoxybenzofuran-2-carboxylate

[0221]

[0222] 4,6-Dimethoxysalicylaldehyde (499.2 mg, 2.74 mmol) was dissolved in DMF (10 mL), followed by the addition of K₂CO₃ (1.89 g, 13.7 mmol) and ethyl bromoethyl (364.8 μL, 3.29 mmol), and reacted at 70 °C for 15 h. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO₄, filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA = 20:1) to obtain 460 mg of the title compound (67% yield, white solid).

[0223] 1 HNMR (400MHz, CDCl3): δ7.54 (m, 1H), 6.67 (m, 1H), 6.35 (d, J=1.9Hz, 1H), 4.41 (q, J=7.2Hz, 2H), 3.90 (s, 3H), 3.857 (s, 3H), 1.40 (t, J=7.2Hz, 3H); (MALDI-TOF): m / z 251[M+H] + ;mp 98℃

[0224] Preparation Example 9: 4,6-Dimethoxybenzofuran-2-carbonylhydrazine

[0225]

[0226] Ethyl 4,6-dimethoxybenzofuran-2-carboxylate (450.5 mg, 1.80 mmol) obtained in Preparation Example 8 was dissolved in EtOH (20 mL), and hydrazine monohydrate (450.5 mg, 9.0 mmol) was added and the mixture was refluxed for 6 hours. The solid formed by filtering the reaction solution and pouring it into cold water was washed with diethyl ether and dried to obtain 360 mg of the title compound (yield 85%, white solid).

[0227] 1 HNMR (400MHz, DMSO-d6): δ9.78 (s, 1H), 7.40 (d, J=0.8Hz, 1H), 6.79 (m, 1H), 6.45 (d, J=2.0Hz, 1H), 4.49 (brs, 2H), 3.88 (s, 3H), 3.82 (s, 3H); 13 CNMR (100MHz, DMSO-d6): δ160.64, 158.10, 156.16, 153.94, 146.15, 110.83, 105.97, 94.98, 88.36, 55.79, 55.71; MS (MALDI-TOF): m / z 259[M+Na] + ;mp 194℃

[0228] Preparation Example 10: Ethyl 5-acetamidobenzofuran-2-carboxylate

[0229]

[0230] Ethyl 5-aminobenzofuran-2-carboxylate (205.2 mg, 1.0 mmol) was dissolved in DMF (3 mL), followed by the addition of (2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethylureon hexafluorophosphate (HBTU, 417.2 mg, 1.1 mmol), hydroxybenzotriazole (HOBt, 148.6 mg, 1.1 mmol), N,N-diisopropylethylamine (DIEA, 209.0 μL, 1.2 mmol), and acetic acid (63.0 μL, 1.1 mmol), and the reaction was carried out at room temperature for 15 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO4, filtered, and distilled under reduced pressure. The residue was separated by column chromatography (CH2Cl2:EA = 3:1) to obtain 162 mg of the title compound (66% yield, white solid).

[0231] 1 HNMR (400MHz, DMSO-d6): δ10.06 (s, 1H), 8.19 (d, J=2.0Hz, 1H), 7.75 (d, J=0.8Hz, 1H), 7.65 (d, J=8 .8Hz, 1H), 7.53 (dd, J=8.8, 2.0Hz, 1H), 4.35 (q, J=7.2Hz, 2H), 2.07 (s, 3H), 1.33 (t, J=7.2Hz, 3H); 13 CNMR (100MHz, DMSO-d6): δ168.76, 159.66, 152.77, 146.68, 134.10, 127.54, 121.15, 114.18, 114.07, 112.66, 61.79, 24.64, 14.49; mp 181-182℃

[0232] Preparation Example 11: N-(2-(hydrazine carbonyl)benzofuran-5-yl)acetamide

[0233]

[0234] Ethyl 5-acetamidobenzofuran-2-carboxylate (514.3 mg, 2.08 mmol) obtained in Preparation Example 10 was dissolved in 1-PrOH (20 mL), and hydrazine monohydrate (312.4 mg, 6.24 mmol) was added and the mixture was refluxed for 24 hours. The solid obtained by vacuum distillation of the reaction solution was washed with a mixed solvent of n-Hex and CH2Cl2 (1:1) and dried to obtain 392 mg of the title compound (yield 81%, white solid).

[0235] 1 HNMR (400MHz, DMSO-d6): δ10.02 (s, 1H), 9.97 (s, 1H), 8.10 (d, J = 1.6Hz, 1H), 7.55 (d, J = 9.2Hz, 1H), 7.48-7.45 (m, 2H), 4.55 (brs, 2H), 2.06 (s, 3H); 13 CNMR (100MHz, DMSO-d6): δ168.23, 157.84, 150.42, 148.93, 135.37, 127.10, 119.12, 111.98, 111.72, 109.06, 23.96; MS (MALDI-TOF) m / z 256[M+Na] + 272[M+K] + ;mp219-220℃

[0236] Preparation of aromatic / N-substituted indole-3-carboxaldehyde derivatives

[0237]

[0238] Preparation Example 12: 2-Chloro-1-methyl-1H-indole-3-carboxaldehyde

[0239]

[0240] At 0 °C, 20 mL of THF was added to 2-chloro-1H-indole-3-carboxaldehyde (359.2 mg, 2.0 mmol) and NaH (120.0 mg, 3.0 mmol, 60% in oil), and the mixture was stirred for 5 minutes. Iodomethane (149.4 μL, 2.4 mmol) was then added, and the mixture was stirred at room temperature for 5 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO4, filtered, and distilled under reduced pressure. The residue was separated by column chromatography (CH2Cl2:EA = 15:1) to obtain 330 mg of the title compound (85% yield, white solid).

[0241] 1 HNMR (400MHz, CDCl3): δ10.13 (s, 1H), 8.30 (m, 1H), 7.36-7.32 (m, 3H), 3.82 (s, 3H); 13 CNMR (100MHz, DMSO-d6): δ184.07, 136.99, 136.13, 124.44, 124.20, 123.69, 121.43, 113.02, 109.64, 30.28; MS (MALDI-TOF): m / z 194[M+H] + ;mp 97-98℃

[0242] Preparation Example 13: Ethyl 2-(2-chloro-3-formyl-1H-indol-1-yl)

[0243]

[0244] At 0 °C, 10 mL of THF was added to 2-chloro-1H-indole-3-carboxaldehyde (200.0 mg, 1.11 mmol) and NaH (66.8 mg, 1.67 mmol, 60% in oil), and the mixture was stirred for 5 minutes. Then, 147.5 μL of ethyl bromoacetate (1.33 mmol) was added, and the mixture was stirred at room temperature for 7 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO4, filtered, and distilled under reduced pressure. The residue was separated by column chromatography (CH2Cl2:EA = 20:1) to obtain 244 mg of the title compound (yield 83%, white solid).

[0245] 1 HNMR (400MHz, CDCl3): δ10.16 (s, 1H), 8.32 (m, 1H), 7.36-7.32 (m, 2H), 7.23 (m, 1H), 4.95 (s, 2H), 4.26 (q, J=7.2Hz, 2H), 1.28 (t, J=7.2Hz, 3H); 13 CNMR (100MHz, CDCl3): δ184.09, 166.47, 136.48, 135.70, 124.45, 124.34, 123.78, 121.55, 113.62, 109.03, 62.36, 44.81, 14.05; MS (MALDI-TOF): m / z 266[M+H] + ;mp110℃

[0246] Preparation Example 14: 2-Chloro-1-(2-ethoxyethyl)-1H-indole-3-carboxaldehyde

[0247]

[0248] 2-Chloro-1H-indole-3-carboxaldehyde (2.69 g, 15.0 mmol) was dissolved in DMF (50 mL), followed by the addition of 2-bromoethyl ether (2.01 mL, 18.0 mmol) and Cs₂CO₃ (14.7 g, 45.0 mmol) and heating at 70 °C for 15 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO₄, filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex∶CH₂Cl₂∶EA=4∶2∶1) to give 2.95 g of the title compound (78% yield, pale yellow solid).

[0249] 1HNMR (400MHz, CDCl3): δ10.14 (s, 1H), 8.30 (m, 1H), 7.41 (m, 1H), 7.34-7.30 (m, 2H), 4.4 2(t, J=5.6Hz, 2H), 3.76 (t, J=5.6Hz, 2H), 3.44 (q, J=7.2Hz, 2H), 1.12 (t, J=7.2Hz, 3H); 13 CNMR (100MHz, CDCl3): δ184.09, 136.58, 135.80, 124.41, 123.95, 123.43, 121.24, 113.08, 110.04, 68.13, 66.91, 43.97, 14.97; MS (MALDI-TOF): m / z 252[M+H] + ;mp52℃

[0250] Preparation Example 15: 2-Chloro-1-(2-methoxyethyl)-1H-indole-3-carboxaldehyde

[0251]

[0252] 2-Chloro-1H-indole-3-carboxaldehyde (538.8 mg, 3.0 mmol) was dissolved in DMF (10 mL), followed by the addition of 2-bromoethyl methyl ether (422.6 μL, 4.5 mmol) and Cs₂CO₃ (2.93 g, 9.0 mmol), and the mixture was heated at 70 °C for 15 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO₄, filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA = 2:1) to obtain 468 mg of the title compound (66% yield, pale yellow solid).

[0253] 1 HNMR (400MHz, CDCl3): δ10.14 (s, 1H), 8.30 (m, 1H), 7.42-7.31 (m, 3H), 4.43 (t, J=5.7Hz, 2H), 3.74 (t, J=5.7Hz, 2H), 3.32 (s, 3H); 13 CNMR (100MHz, CDCl3): δ184.27, 136.79, 135.97, 124.56, 124.21, 123.64, 121.44, 113.25, 110.17, 70.49, 59.35, 44.00; MS (MALDI-TOF): m / z 238[M+H] + 260[M+Na] + ;mp 66-67℃

[0254] Preparation Example 16: 5-Methyloxyindole

[0255]

[0256] Ethylene glycol (10 mL) was added to 5-methylindigo (1.50 g, 9.31 mmol), followed by the addition of KOH (522.4 mg, 9.31 mmol) and hydrazine monohydrate (1.40 g, 27.9 mmol), and the mixture was heated at 140 °C for 4 hours. The reaction solution was cooled and acidified with 1 N HCl, followed by extraction with EA. The organic layer was subjected to vacuum distillation, and the residue was separated by column chromatography (n-Hex:EA = 3:2) to give 1.0 g of the title compound (73% yield, light brown solid).

[0257] 1 HNMR (400MHz, DMSO-d6): δ10.23 (brs, 1H), 7.01 (s, 1H), 6.96 (d, J=7.9Hz, 1H), 6.69 (d, J=7.9Hz, 1H), 3.41 (s, 2H), 2.23 (s, 3H)

[0258] Preparation Example 17: 2-Chloro-5-methyl-1H-indole-3-carboxaldehyde

[0259]

[0260] At 0°C, POCl3 (2.65 mL, 28.9 mmol) was added to 10 mL of DMF and stirred for 10 minutes. Then, a 10 mL solution of 5-methyloxyindole (850.0 mg, 5.78 mmol) in DMF was added and the mixture was heated at 80°C for 3 hours. After alkalization with 1 N NaOH, the reaction solution was extracted with EA. The organic layer was washed with water, dried over Na2SO4, filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA = 3:2) to obtain 600 mg of the title compound (54% yield, pale yellow solid).

[0261] 1 HNMR (400MHz, DMSO-d6): δ12.96 (s, 1H), 9.96 (s, 1H), 7.87 (s, 1H), 7.31 (d, J=8.3Hz), 7.10 (dd, J=8.3, 1.5Hz, 1H), 2.39 (s, 3H); mp 215℃

[0262] Preparation Example 18: 2-Chloro-1-(2-ethoxyethyl)-5-methyl-1H-indole-3-carboxaldehyde

[0263]

[0264] A solution of 2-chloro-5-methyl-1H-indole-3-carboxaldehyde (440.0 mg, 2.27 mmol) in CH3CN (20 mL) was added with 303.8 μL of 2.72 mmol and Cs2CO3 (3.71 g, 11.4 mmol) and refluxed for 15 h. The reaction solution was distilled under reduced pressure, and the residue was extracted with EA after adding water. The residue was dried over MgSO4, filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA = 9:1) to give 570 mg of the title compound (94%, white solid).

[0265] 1 HNMR (400MHz, CDCl3): δ10.09 (s, 1H), 8.10 (s, 1H), 7.28 (d, J=8.5Hz, 1H), 7.14 (d, J=8.5, 1.5Hz, 1H), 4 .39 (t, J=5.5Hz, 2H), 3.74 (t, J=5.5Hz, 2H), 3.43 (q, J=7.0Hz, 2H), 2.46 (s, 3H), 1.11 (t, J=7.0Hz, 3H); 13 CNMR (400MHz, DMSO-d6): δ184.35, 136.69, 134.28, 133.48, 125.59, 124.73, 12 1.19, 112.92, 109.89, 68.30, 67.09, 44.19, 21.61, 15.17; MS (MALDI-TOF): m / z 266[M+H] + ;mp54℃

[0266] Preparation Example 19: 2-Chloro-5-methoxy-1H-indole-3-carboxaldehyde

[0267]

[0268] At 0°C, POCl3 (2.24 mL g, 24.5 mmol) was added to 10 mL of DMF and stirred for 10 minutes. Then, a 10 mL solution of 5-methoxyoxyindole (1.60 g, 9.81 mmol) in DMF was added, and the mixture was heated at 80°C for 2 hours. After alkalization with 1 N NaOH, the resulting solid was washed with water and dried to obtain the title compound. The filtrate was extracted again with EA, dried with Na2SO4, filtered, and then distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA = 1:2) to obtain 1.27 g of the title compound (62% yield, light brown solid).

[0269] 1 HNMR (300MHz, DMSO-d6): δ12.96 (brs, 1H), 9.96 (s, 1H), 7.57 (d, J=2.7Hz, 1H), 7.33 (d, J=8.7Hz, 1H), 6.90 (dd, J=8.7, 2.7Hz, 1H), 3.79 (s, 3H); mp 225℃

[0270] Preparation Example 20: 2-Chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indole-3-carboxaldehyde

[0271]

[0272] 5-Methoxy-2-chloro-1H-indole-3-carboxaldehyde (1.97 g, 9.40 mmol) was dissolved in DMF (70 mL), followed by the addition of 2-bromoethyl ether (1.26 mL, 11.3 mmol) and Cs₂CO₃ (15.3 g, 47.0 mmol) and heating at 70 °C for 11 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO₄, filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA = 3:1) to give 2.18 g of the title compound (82% yield, pale yellow solid).

[0273] 1 HNMR (400MHz, CDCl3): δ10.09 (s, 1H), 7.79 (d, J=2.7Hz, 1H), 7.29 (d, J=9.0Hz, 1H), 6.94 (dd, J=9.0, 2.7Hz, 1H), 4.37 (t, J=5.7Hz, 2H), 3.89 (s, 3H), 3.74 (t, J=5.7Hz, 2H), 3.43 (q, J=7.2Hz, 2H), 1.11 (t, J=7.2Hz, 3H); 13 CNMR (100MHz, CDCl3): δ184.10, 156.90, 135.99, 130.59, 125.12, 114.17, 112 .95, 111.06, 102.64, 68.21, 66.90, 55.77, 44.16, 14.96; MS (MALDI-TOF): m / z 282[M+H] + ;mp 48℃

[0274] Preparation Example 21: 6-Methoxyoxyindole

[0275]

[0276] Ethylene glycol (10 mL) was added to 6-methoxyindigo (500 mg, 2.82 mmol), followed by the addition of KOH (158.2 mg, 2.82 mmol) and hydrazine monohydrate (282.3 mg, 5.64 mmol), and the mixture was heated at 140 °C for 4 hours. The reaction solution was cooled and acidified with 1 N HCl, then extracted with EA. The organic layer was subjected to vacuum distillation, and the residue was separated by column chromatography (n-Hex:EA = 1:1) to obtain 278 mg of the title compound (60% yield, pale yellow solid).

[0277] 1 HNMR (400MHz, DMSO-d6): δ10.30 (brs, 1H), 7.08 (d, J=8.0Hz, 1H), 6.48 (d, J=8.0, 2.4Hz, 1H), 6.38 (d, J=2.4Hz, 1H), 3.71 (s, 3H), 3.37 (s, 2H)

[0278] Preparation Example 22: 2-Chloro-6-methoxy-1H-indole-3-carboxaldehyde

[0279]

[0280] At 0°C, POCl3 (1.79 mL g, 19.5 mmol) was added to DMF (5 mL) and stirred for 10 minutes. Then, a DMF (15 mL) solution of 6-methoxyoxyindole (1.27 g, 7.78 mmol) was added and the mixture was heated at 80°C for 2 hours. After alkalization with 1 N NaOH, the resulting solid was filtered, washed with water, and dried to obtain the title compound. The filtrate was extracted again with EA, dried with Na2SO4, filtered, and then distilled under reduced pressure. The residue was separated by column chromatography (CH2Cl2:EA = 10:1) to obtain 945 mg of the title compound (yield, white solid).

[0281] 1 HNMR (400MHz, DMSO-d6): δ12.87 (brs, 1H), 9.94 (s, 1H), 7.91 (m, 1H), 6.89-6.87 (m, 2H), 3.79 (s, 3H); 13 CNMR (100MHz, DMSO-d6): δ183.09, 156.90, 135.57, 133.26, 120.73, 118.09, 112.23, 112.10, 95.09, 55.31; mp 230℃

[0282] Preparation Example 23: 2-Chloro-1-(2-ethoxyethyl)-6-methoxy-1H-indole-3-carboxaldehyde

[0283]

[0284] 6-Methoxy-2-chloro-1H-indole-3-carboxaldehyde (765.1 mg, 3.65 mmol) was dissolved in DMF (15 mL), followed by the addition of 2-bromoethyl ether (489.2 μL, 4.38 mmol) and Cs₂CO₃ (3.58 g, 11.0 mmol), and the mixture was heated at 70 °C for 5 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO₄, filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex∶CH₂Cl₂∶EA=3∶1∶0.5) to obtain 420 mg of the title compound (41% yield, white solid).

[0285] 1 HNMR (400MHz, CDCl3): δ10.08 (s, 1H), 8.15 (d, J=8.8Hz, 1H), 6.95 (dd, J=8.8, 2.4Hz, 1H), 6.89 (d, J=2.4Hz, 1H), 4.36 (t, J=5.6Hz, 2H), 3.87 (s, 3H), 3.75 (t, J=5.6Hz, 2H), 3.45 (q, J=7.2Hz, 2H), 1.13 (t, J=7.2Hz, 3H); 13 CNMR (100MHz, DMSO-d6): δ184.02, 157.51, 136.85, 135.25, 122.00, 118.29, 1 13.20, 112.31, 94.46, 68.25, 66.92, 55.68, 44.00, 15.04; MS (MALDI-TOF): m / z 281[M] + ;mp 82℃

[0286] Preparation Example 24: 2-Chloro-5-fluoro-1H-indole-3-carboxaldehyde

[0287]

[0288] At 0 °C, POCl3 (1.36 mL g, 14.9 mmol) was added to DMF (5 mL) and stirred for 10 minutes. Then, a DMF (15 mL) solution of 5-fluorooxyindole (900.0 mg, 5.95 mmol) was added and the mixture was heated at 80 °C for 4 hours. After alkalization with 1 N NaOH, the reaction solution was extracted with EA. The organic layer was dried over Na2SO4, filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA = 1:1) to obtain 235 mg of the title compound (20% yield, white solid).

[0289] 1 HNMR (300MHz, DMSO-d6): δ13.22 (brs, 1H), 9.96 (s, 1H), 7.73 (dd, J=9.0, 2.7Hz, 1H), 7.45 (dd, J=9.0, 4.5Hz, 1H), 7.13 (m, 1H); 13 CNMR (100MHz, DMSO-d6): δ183.27, 158.93 (d, J=235.2Hz), 135.60, 131.21, 124.89 (d, J=11.3H z), 113.27 (d, J=9.9Hz), 112.11 (d, J=4.4Hz), 111.92 (d, J=25.9Hz), 105.09 (d, J=25.1Hz); mp 208-210℃

[0290] Preparation Example 25: 2-Chloro-1-(2-ethoxyethyl)-5-fluoro-1H-indole-3-carboxaldehyde

[0291]

[0292] 2-Chloro-5-fluoro-1H-indole-3-carboxaldehyde (197.6 mg, 1.0 mmol) was dissolved in DMF (5 mL), followed by the addition of 2-bromoethyl ether (134.0 μL, 1.2 mmol) and Cs₂CO₃ (977.5 mg, 3.0 mmol) and heating at 70 °C for 15 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO₄, filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex∶CH₂Cl₂∶EA=3∶1∶0.5) to give 125 mg of the title compound (46% yield, white solid).

[0293] 1 HNMR (400MHz, CDCl3): δ10.09 (s, 1H), 7.98 (d, J=9.2, 2.4Hz, 1H), 7.36 (d, J=9.2, 4.4Hz, 1H), 7.06 (m, 1H), 4.41 (t, J = 5.6Hz, 2H), 3.76 (t, J = 5.6Hz, 2H), 3.43 (q, J = 7.2Hz, 2H), 1.11 (t, J = 7.2Hz, 3H); 13CNMR (400MHz, DMSO-d6): δ183.79, 159.96 (d, J=238.6Hz), 137.13, 132.38, 124.95 (d, J=11.3Hz), 113.07 (d, J=4.4Hz ), 112.19 (d, J = 26.2Hz), 111.30 (d, J = 9.4Hz), 106.80 (d, J = 25.1Hz), 68.23, 66.91, 44.37, 14.93; MS (MALDI-TOF): m / z 270[M+H] + ;mp 83-84℃

[0294] Preparation Example 26: 2,5-Dichloro-1H-indole-3-carboxaldehyde

[0295]

[0296] At 0°C, POCl3 (2.74 mL, 29.9 mmol) was added to DMF (5 mL) and stirred for 10 minutes. Then, a DMF (5 mL) solution of 5-chlorooxyindole (1.0 g, 5.97 mmol) was added and the mixture was heated at 80°C for 3 hours. The reaction solution was alkalized by adding 1 N NaOH and extracted with EA. The organic layer was washed with water, dried over Na2SO4, filtered, and then distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA = 2:1) to obtain 530 mg of the title compound (41% yield, light brown solid).

[0297] 1 HNMR (400MHz, DMSO-d6): δ13.31 (brs, 1H), 9.97 (s, 1H), 8.04 (d, J=2.0Hz, 1H), 7.47 (d, J=8.5Hz, 1H), 7.31 (dd, J=8.5, 2.0Hz, 1H); mp 245-248℃

[0298] Preparation Example 27: 2,5-Dichloro-1-(2-ethoxyethyl)-1H-indole-3-carboxaldehyde

[0299]

[0300] A solution of 2,5-dichloro-1H-indole-3-carboxaldehyde (350 mg, 1.64 mmol) in CH3CN (20 mL) obtained in Preparation Example 26 above was added with 2-bromoethyl ether (274.8 μL, 2.46 mmol) and Cs2CO3 (2.67 g, 8.20 mmol) and refluxed for 15 hours. The reaction solution was distilled under reduced pressure, and water was added to the residue, followed by extraction with EA. The residue was dried over MgSO4, filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA = 9:1) to obtain 360 mg of the title compound (77% yield, pale yellow solid).

[0301] 1 HNMR (400MHz, CDCl3): δ10.06 (s, 1H), 8.26 (d, J=2.0Hz, 1H), 7.32-7.23 (m, 2H), 4.38 (t, J=5.6Hz, 2H), 3.73 (t, J=5.6Hz, 2H), 3.41 (q, J=7.2Hz, 2H), 1.08 (t, J=7.2Hz, 3H); 13 CNMR (100MHz, DMSO-d6): δ184.01, 137.39, 134.50, 129.64, 125.40, 124.51, 120 .95, 112.84, 111.60, 68.41, 67.13, 44.55, 15.15; MS (MALDI-TOF): m / z286[M+H] + ;mp 104℃

[0302] Preparation Example 28: 5-(trifluoromethoxy)oxyindole

[0303]

[0304] Ethylene glycol (10 mL) was added to 5-(trifluoromethoxy)indigo (1.75 g, 7.57 mmol), followed by the addition of KOH (424.8 mg, 7.57 mmol) and hydrazine monohydrate (1.14 g, 22.7 mmol), and the mixture was heated at 140 °C for 4 hours. The reaction solution was cooled, acidified with 1 N HCl, and extracted with EA. The organic layer was subjected to vacuum distillation, and the residue was separated by column chromatography (n-Hex:EA = 1:2) to obtain 855 mg of the title compound (52% yield, light brown solid).

[0305] 1HNMR (400MHz, DMSO-d6): δ10.52 (brs, 1H), 7.24 (s, 1H), 7.16 (m, 1H), 6.86 (d, J=8.0Hz, 1H), 3.54 (s, 2H)

[0306] Preparation Example 29: 2-Chloro-5-(trifluoromethoxy)-1H-indole-3-carboxaldehyde

[0307]

[0308] At 0 °C, POCl3 (727.7 μL g, 7.95 mmol) was added to 1 mL of DMF and stirred for 10 minutes. Then, a 5 mL solution of 5-(trifluoromethoxy)oxyindole (575.0 mg, 2.65 mmol) in DMF was added, and the mixture was heated at 80 °C for 3 hours. The reaction solution was alkalized by adding 1 N NaOH and extracted with EA. The organic layer was dried over Na2SO4, filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA = 1:1) to obtain 120 mg of the title compound (17% yield, white solid).

[0309] 1 HNMR (400MHz, DMSO-d6): δ13.40 (brs, 1H), 9.99 (s, 1H), 7.95 (m, 1H), 7.54 (d, J=8.8Hz, 1H), 7.29 (m, 1H); mp191-192℃

[0310] Preparation Example 30: 2-Chloro-1-(2-ethoxyethyl)-5-(trifluoromethoxy)-1H-indole-3-carboxaldehyde

[0311]

[0312] 2-Chloro-5-(trifluoromethoxy)-1H-indole-3-carboxaldehyde (100 mg, 0.38 mmol) was dissolved in DMF (3 mL), followed by the addition of 2-bromoethyl ether (51.4 μL, 0.46 mmol) and Cs₂CO₃ (371.4 mg, 1.14 mmol) and heating at 70 °C for 8 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO₄, filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:CH₂Cl₂:EA = 4:4:1) to obtain 43 mg of the title compound (34% yield, pale yellow solid).

[0313] 1HNMR (400MHz, CDCl3): δ10.11 (s, 1H), 8.18 (s, 1H), 7.42 (d, J=8.8Hz, 1H), 7.20 (dd, J=8.8, 1.6Hz , 1H), 4.42 (t, J=5.6Hz, 2H), 3.77 (t, J=5.6Hz, 2H), 3.44 (q, J=5.2Hz, 2H), 1.11 (t, J=7.2Hz, 3H); 13 CNMR (100MHz, CDCl3): δ183.78, 145.66 (q, J=1.9Hz), 137.58, 134.20, 124.73, 120.62 (q, J=255.0H z), 117.81, 113.86 (q, J=0.8Hz), 113.23, 111.29, 68.25, 66.96, 44.46, 14.94; MS (MALDI-TOF): m / z 335[M] + ;mp 79℃

[0314] Preparation Example 31: 2-Bromo-1H-indole-3-carboxaldehyde

[0315]

[0316] At 0 °C, DMF (1.8 mL) was added to CH₂Cl₂ (6 mL), followed by the slow addition of a CH₂Cl₂ (10 mL) solution of POBr₃ (5.33 g, 18.6 mmol) and reflux for 15 min. Then, oxyindole (1.03 g, 7.74 mmol) was gradually added and refluxed again for 1 h. The reaction solution was then added to cold water and stirred for 20 min, thereby separating the aqueous layer. The aqueous layer was neutralized with solid K₂CO₃, and the resulting solid was filtered. The obtained solid was purified by column chromatography (n-Hex:EA = 2:1) to give 1.2 g of the title compound (70% yield, light brown solid).

[0317] 1 HNMR (400MHz, DMSO-d6): δ13.04 (brs, 1H), 9.90 (s, 1H), 8.08 (m, 1H), 7.43 (m, 1H), 7.29-7.21 (m, 2H)

[0318] Preparation Example 32: Ethyl 2-(2-bromo-3-formyl-1H-indol-1-yl)

[0319]

[0320] At 0°C, 2-bromo-1H-indole-3-carboxaldehyde (120.0 mg, 0.54 mmol) and NaH (32.4 mg, 0.81 mmol, 60% in oil) obtained in Preparation Example 31 above were mixed with THF (5 mL) and DMF (2 mL) and stirred for 5 minutes. Then, ethyl bromoacetate (72.1 μL, 0.65 mmol) was added and stirred at room temperature for 15 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO4, filtered, and distilled under reduced pressure. The residue was separated by column chromatography (CH2Cl2:EA = 20:1) to obtain 91 mg of the title compound (54% yield, white solid).

[0321] 1 HNMR (400MHz, CDCl3): δ10.07 (s, 1H), 8.33 (m, 1H), 7.34-7.31 (m, 2H), 7.25 (m, 1H), 4.99 (s, 2H), 4.26 (q, J=7.2Hz, 2H), 1.28 (t, J=7.2Hz, 3H); 13 CNMR (100MHz, CDCl3): δ185.45, 166.55, 137.11, 126.09, 125.18, 124.49, 123.62, 121.40, 116.07, 109.21, 62.35, 46.27, 14.06; MS (MALDI-TOF): m / z 309[M] + ;mp94℃

[0322] Preparation Example 33: 2-Bromo-1-(2-ethoxyethyl)-1H-indole-3-carboxaldehyde

[0323]

[0324] 2-Bromo-1H-indole-3-carboxaldehyde (400.0 mg, 1.79 mmol) obtained in Preparation Example 31 above was dissolved in DMF (10 mL), followed by the addition of 2-bromoethyl ether (240.1 μL, 2.15 mmol) and Cs₂CO₃ (1.75 g, 5.37 mmol), and heated at 70 °C for 6 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO₄, filtered, and distilled under reduced pressure. The residue was separated by column chromatography (CH₂Cl₂:EA = 20:1) to obtain 436 mg of the title compound (yield 82%, white solid).

[0325] 1HNMR (400MHz, CDCl3): δ10.06 (s, 1H), 8.32 (m, 1H), 7.43 (m, 1H), 7.34-7.30 (m, 2H), 4.4 6 (t, J=5.6Hz, 2H), 3.77 (t, J=5.6Hz, 2H), 3.45 (q, J=7.2Hz, 2H), 1.23 (t, J=7.2Hz, 3H); 13 CNMR (100MHz, CDCl3): δ185.61, 137.43, 126.59, 125.43, 124.25, 123.57, 121.30, 115.60, 110.47, 68.40, 67.14, 45.59, 15.17; MS (MALDI-TOF): m / z 295[M] + ;mp 56-57℃

[0326] Preparation Example 34: 2-(trifluoromethyl)-1H-indole-3-carboxaldehyde

[0327]

[0328] At 0°C, POCl3 (1.83 mL g, 20.0 mmol) was added to DMF (10 mL) and stirred for 10 minutes. Then, a DMF (10 mL) solution of 2-trifluoromethylindole (740.6 mg, 4.0 mmol) was added, and the mixture was heated at 80°C for 5 hours. The reaction solution was alkalized by adding 1 N NaOH and extracted with EA. The organic layer was washed with brine, dried over Na2SO4, filtered, and then distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA = 7:1) to obtain 324 mg of the title compound (38% yield, white solid).

[0329] 1 HNMR (400MHz, DMSO-d6): δ13.42 (brs, 1H), 10.24 (s, 1H), 8.25 (d, J=8.0Hz, 1H), 7.61 (d, J=8.0Hz, 1H), 7.44 (m, 1H), 7.36 (m, 1H); mp 171-173℃

[0330] Preparation Example 35: 1-(2-ethoxyethyl)-2-(trifluoromethyl)-1H-indole-3-carboxaldehyde

[0331]

[0332] 2-(trifluoromethyl)-1H-indole-3-carboxaldehyde (350.0 mg, 1.64 mmol) was dissolved in DMF (10 mL), followed by the addition of 2-bromoethyl ether (220.0 μL, 1.97 mmol) and Cs₂CO₃ (1.60 g, 4.92 mmol), and the mixture was heated at 70 °C for 15 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO₄, filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA = 5:1) to give 184 mg of the title compound (39% yield, white solid).

[0333] 1 HNMR (400MHz, CDCl3): δ10.41 (s, 1H), 8.52 (d, J=8.0Hz, 1H), 7.58 (d, J=8.0Hz, 1H), 7.46 (m, 1H), 7.3 9 (m, 1H), 4.52 (t, J=6.0Hz, 2H), 3.80 (t, J=6.0Hz, 2H), 3.45 (q, J=7.2Hz, 2H), 1.13 (t, J=7.2Hz, 3H); 13 CNMR (100MHz, CDCl3): δ185.81, 137.63, 131.27 (q, J=37.9Hz), 126.02, 124.30, 124.21, 123.66, 121.29 (q , J=270.3Hz), 117.84 (q, J=1.5Hz), 111.18, 68.82, 66.93, 45.58 (q, J=2.5Hz), 14.94; MS (MALDI-TOF): m / z 286[M+H] + Preparation of benzofuranyl N-acylhydrazone derivatives at 46℃

[0334]

[0335] Example 1: Preparation of (E)-N'-[(2-chloro-1H-indol-3-yl)methylene]-5-methylbenzofuran-2-carbonylhydrazine (CAP-1052, compound 7)

[0336]

[0337] 1-PrOH (15 mL) and acetic acid (1-2 drops) were added to 5-methylbenzofuran-2-carbonylhydrazine (95.1 mg, 0.50 mmol) and 2-chloro-1H-indole-3-carboxaldehyde (89.8 mg, 0.50 mmol), and the mixture was refluxed for 8 hours. The reaction solution was distilled under reduced pressure, and the residue was washed with a 1:1 mixture of n-Hex / CH₂Cl₂ to give 153 mg of the title compound (87% yield, pale yellow solid).

[0338] 1 HNMR (300MHz, DMSO-d6) δ12.48 (s, 1H), 11.98 (s, 1H), 8.73 (s, 1H), 8.28 (d, J=7.2Hz, 1H), 7.6 3-7.58 (m, 3H), 7.39 (d, J=7.2Hz, 1H), 7.32 (d, J=8.8Hz, 1H), 7.27-7.19 (m, 2H), 2.44 (s, 3H); 13 CNMR (100MHz, DMSO-d6) δ154.20, 152.86, 148.41, 142.86, 135.02, 132.91, 128.33, 127.26, 127.1 6, 124.06, 123.20, 122.24, 121.33, 121.28, 111.37, 111.21, 110.03, 107.39, 20.83; HRMS (TOFMSES - ): m / zcalcdforC 19 H 13 ClN3O2(MH) - 350.0696, found350.0700;mp 242℃

[0339] Example 2: Preparation of (E)-N'-[(2-chloro-1-methyl-1H-indol-3-yl)methylene]-5-methylbenzofuran-2-carbonylhydrazine (CAP-1041, compound 13)

[0340]

[0341] 1-PrOH (15 mL) and acetic acid (1-2 drops) were added to 5-methylbenzofuran-2-carbonylhydrazine (95.1 mg, 0.50 mmol) and 2-chloro-1-methyl-1H-indole-3-carboxaldehyde (96.8 mg, 0.50 mmol), and the mixture was refluxed for 24 hours. After cooling the reaction solution and filtering the solid, 100 mg of the title compound (55% yield, light brown solid) was obtained by washing with EtOH.

[0342] 1HNMR (400MHz, DMSO-d6): δ12.00 (s, 1H), 8.75 (s, 1H), 8.32 (d, J=7.6Hz, 1H), 7.63-7.57 (m, 4H), 7.35-7.25 (m, 3H), 3.81 (s, 3H), 2.43 (s, 3H); 13 CNMR (100MHz, DMSO-d6): δ154.22, 152.87, 148.40, 142.93, 136.08, 132.92, 129.29, 128.34, 127.16, 1 23.24, 123.21, 122.25, 121.67, 121.42, 111.38, 110.34, 110.06, 107.28, 30.19, 20.84; HRMS (TOFMSES - ): m / zcalcdforC 20 H 15 ClN3O2(MH) - 364.0853, found 364.0847; mp 246-247℃

[0343] Example 3: Preparation of ethyl (E)-2-{2-chloro-3-[(2-(5-methylbenzofuran-2-carbonyl)hydrazine)methyl]-1H-indol-1-yl} (CAP-1038, compound 12)

[0344]

[0345] 1-PrOH (15 mL) and acetic acid (1-2 drops) were added to 5-methylbenzofuran-2-carbonylhydrazine (60.9 mg, 0.32 mmol) and ethyl 2-(2-chloro-3-formyl-1H-indol-1-yl)acetate (85.0 mg, 0.32 mmol), and the mixture was refluxed for 30 hours. The reaction solution was distilled under reduced pressure, and the residue was washed with Et₂O to give 85 mg of the title compound (61% yield, white solid).

[0346] 1 HNMR (400MHz, DMSO-d6): δ12.06 (s, 1H), 8.76 (s, 1H), 8.34 (d, J=6.8Hz, 1H), 7.64-7.58 (m, 4H ), 7.34-7.27 (m, 3H), 5.26 (s, 2H), 4.19 (q, J=7.2Hz, 2H), 2.43 (s, 3H), 1.22 (t, J=7.2Hz, 3H); 13CNMR (100MHz, DMSO-d6): δ167.84, 154.28, 152.88, 148.33, 142.65, 136.14, 132.94, 129.17, 128.39, 127.15, 123 .57, 123.29, 122.27, 121.97, 121.49, 111.39, 110.30, 110.19, 108.22, 61.45, 44.78, 20.84, 14.00; HRMS (TOFMSES - ): m / zcalcdforC 23 H 19 ClN3O4(MH) - 436.1064, found 436.1068; mp 198℃

[0347] Example 4: Preparation of (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbonylhydrazine (CAP-1042, compound 1)

[0348]

[0349] 1-PrOH (15 mL) and acetic acid (1-2 drops) were added to 5-methylbenzofuran-2-carbazide (98.9 mg, 0.52 mmol) and 2-chloro-1-(2-ethoxyethyl)-1H-indole-3-carboxaldehyde (130.9 mg, 0.52 mmol), and the mixture was refluxed for 24 hours. The reaction solution was distilled under reduced pressure, and the residue was separated by column chromatography (n-Hex:EA = 1:1) to give 179 mg of the title compound (yield 81%, pale yellow solid).

[0350] 1 HNMR (400MHz, DMSO-d6): δ12.01 (s, 1H), 8.75 (s, 1H), 8.32 (d, J=7.6Hz, 1H), 7.63-7.58 (m, 4H), 7.33-7.24 (m , 3H), 4.46 (t, J=5.6Hz, 2H), 3.71 (t, J=5.6Hz, 2H), 3.39 (q, J=6.8Hz, 2H), 2.43 (s, 3H), 1.00 (t, J=6.8Hz, 3H); 13CNMR (100MHz, DMSO-d6): δ154.23, 152.87, 148.40, 142.99, 135.76, 132.92, 129.26, 128.34, 127, 16, 123.35, 123 .21, 122.25, 121.67, 121.43, 111.37, 110.67, 110.08, 107.53, 67.85, 65.63, 43.52, 20.83, 14.90; HRMS (TOFMSES - ): m / zcalcdf orC 23 H 21 ClN3O3(MH) - 422.1271, found 422.1261;mp 172℃

[0351] Example 5: Preparation of (E)-N'-{[2-bromo-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbonylhydrazine (CAP-1044, compound 14)

[0352]

[0353] 1-PrOH (15 mL) and acetic acid (1-2 drops) were added to 5-methylbenzofuran-2-carbazide (108.4 mg, 0.57 mmol) and 2-bromo-1-(2-ethoxyethyl)-1H-indole-3-carboxaldehyde (168.8 mg, 0.57 mmol), and the mixture was refluxed for 24 hours. The reaction solution was distilled under reduced pressure and dissolved in a small amount of CH2Cl2, then added dropwise to an n-Hex solution. 209 mg of the title compound was obtained by filtration of the resulting solid (78% yield, pale yellow solid).

[0354] 1 HNMR (400MHz, DMSO-d6): δ12.03 (s, 1H), 8.72 (s, 1H), 8.35 (d, J=7.2Hz, 1H), 7.63-7.58 (m, 4H), 7.33-7.22 (m , 3H), 4.48 (t, J=5.6Hz, 2H), 3.71 (t, J=5.6Hz, 2H), 3.39 (q, J=7.2H, 2H), 2.44 (s, 3H), 1.01 (t, J=7.2Hz, 3H); 13CNMR (100MHz, DMSO-d6): δ154.21, 152.86, 148.41, 144.36, 137.00, 132.90, 128.32, 127.16, 124.14, 123.13, 122 .23, 121.46, 121.29, 119.80, 111.36, 110.77, 110.43, 110.03, 67.96, 65.69, 44.81, 20.83, 14.91; HRMS (TOFMSES - ): m / zcalcdforC 23 H 21 BrN3O3(MH) - 466.0766, found 466.0776;mp 190℃

[0355] Example 6: Preparation of (E)-N'-{[1-(2-ethoxyethyl)-2-(trifluoromethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbazide (CAP-1045, compound 15)

[0356]

[0357] 1-PrOH (15 mL) and acetic acid (1-2 drops) were added to 5-methylbenzofuran-2-carbazide (93.2 mg, 0.49 mmol) and 1-(2-ethoxyethyl)-2-trifluoromethyl-1H-indole-3-carboxaldehyde (139.8 mg, 0.49 mmol), and the mixture was refluxed for 24 hours. The reaction solution was distilled under reduced pressure, and the residue was separated by column chromatography (n-Hex:EA = 2:1) to give 176 mg of the title compound (79% yield, white solid).

[0358] 1 HNMR (400MHz, DMSO-d6): δ12.24 (s, 1H), 9.00 (s, 1H), 8.63 (d, J=8.0Hz, 1H), 7.75 (d, J=8.4Hz, 1H), 7.67 (s, 1H), 7.61-7.59 (m, 2H), 7. 47 (m, 1H), 7.37-7.32 (m, 2H), 4.54 (t, J=5.4Hz, 2H), 3.72 (t, J=5.4Hz, 2H), 3.37 (q, J=7.0Hz, 2H), 2.44 (s, 3H), 10.00 (t, J=7.0Hz, 3H); 13CNMR (100MHz, DMSO-d6): δ154.52, 152.93, 148.17, 142.47, 137.83, 133.01, 128.53, 127.12, 125.66, 125.10 (q, J=35.8Hz), 1 24.20, 123.14, 122.48, 122.32, 121.75 (q, J=269.0Hz), 113.45, 111.85, 111.42, 110.56, 68.36, 65.79, 45.01, 20.84, 14.85; 19 FNMR (376MHz, DMSO-d6): δ-52.9 (s, 3F); HRMS (TOFMSES - ): m / zcalcdforC 24 H 21 F3N3O3(MH) - 456.1535, found 456.1531; mp 183℃

[0359] Example 7: Preparation of (E)-N'-{[2-chloro-1-(2-methoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbonylhydrazine (CAP-1070, compound 19)

[0360]

[0361] 1-PrOH (20 mL) and acetic acid (1-2 drops) were added to 5-methylbenzofuran-2-carbazide (319.5 mg, 1.68 mmol) and 2-chloro-1-(2-methoxyethyl)-1H-indole-3-carboxaldehyde (399.3 mg, 1.68 mmol), and the mixture was refluxed for 15 hours. After cooling the reaction solution and filtering the solid, 630 mg of the title compound (91% yield, pale yellow solid) was obtained by washing with n-Hex.

[0362] 1 HNMR (400MHz, DMSO-d6): δ12.01 (s, 1H), 8.75 (s, 1H), 8.33 (d, J=7.5Hz, 1H), 7.63-7.58 (m, 4H ), 7.33-7.24 (m, 3H), 4.48 (t, J = 5.4Hz, 2H), 3.68 (t, J = 5.4Hz, 2H), 3.21 (s, 3H), 2.43 (s, 3H); 13CNMR (100MHz, DMSO-d6): δ154.23, 152.88, 148.40, 142.97, 135.76, 132.93, 129.20, 128.36, 127.16, 123.34, 123.25, 122.25, 121.68, 121.44, 111.38, 110.68, 110.10, 107.55, 70.11, 58.29, 43.34, 20.84; HRMS (TOFMSES - ): m / zcalcdforC 22 H 19 ClN3O3(MH) - 408.1115, found 408.1099; mp 243℃

[0363] Example 8: Preparation of (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbonylhydrazine (CAP-1046, compound 2)

[0364]

[0365] 1-PrOH (15 mL) and acetic acid (1-2 drops) were added to 5-methylbenzofuran-2-carbazide (127.4 mg, 0.67 mmol) and 2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indole-3-carboxaldehyde (188.8 mg, 0.67 mmol), and the mixture was refluxed for 24 hours. The reaction solution was subjected to vacuum distillation, and the residue was separated by column chromatography (n-Hex:EA = 1:2) to obtain 213 mg of the title compound (70% yield, pale yellow solid).

[0366] 1 HNMR (400MHz, DMSO-d6): δ12.01 (s, 1H), 8.73 (s, 1H), 7.91 (d, J=2.4Hz, 1H), 7.61-7.57 (m, 3H), 7.52 (d, J=9.2Hz, 1H), 7.32 (dd, J=8.8, 1.6Hz, 1H) , 6.95 (dd, J=8.8, 2.4Hz, 1H), 4.42 (t, J=5.2Hz, 2H), 3.82 (s, 3H), 3.68 (t , J=5.2Hz, 2H), 3.38 (q, J=7.2Hz, 2H), 2.43 (s, 3H), 1.00 (t, J=7.2Hz, 3H); 13CNMR (400MHz, DMSO-d6): δ155.20, 154.17.152.85, 148.48, 142.94, 132.91, 130.74, 128.95, 128.31, 127.17, 124.01 , 122.23, 112.24, 111.53, 111.35, 110.04, 107.24, 104.11, 67.91, 65.62, 55.33, 43.65, 20.83, 14.90; HRMS (TOFMSES - ): m / zcalcdforC 24 H 23 ClN3O4(MH)<->452.1377, foun d 452.1355; mp 208℃

[0367] Example 9: Preparation of (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-6-methoxy-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbonylhydrazine (CAP-1050, compound 6)

[0368]

[0369] 1-PrOH (15 mL) and acetic acid (1-2 drops) were added to 5-methylbenzofuran-2-carbazide (95.1 mg, 0.50 mmol) and 2-chloro-1-(2-ethoxyethyl)-6-methoxy-1H-indole-3-carboxaldehyde (140.9 mg, 0.50 mmol), and the mixture was refluxed for 16 hours. The reaction solution was distilled under reduced pressure and dissolved in a small amount of HCl, then added dropwise to an n-Hex solution. The solid obtained was filtered to give 205 mg of the title compound (90% yield, pale yellow solid).

[0370] 1 HNMR (400MHz, DMSO-d6): δ11.98 (s, 1H), 8.70 (s, 1H), 8.16 (d, J=8.8Hz, 1H), 7.62-7.58 (m, 3H), 7.32 (dd, J=8.4, 1.2Hz, 1H), 7.17 (d, J=2.0Hz, 1H) , 6.90 (dd, J=8.8, 2.4Hz, 1H), 4.42 (t, J=5.6Hz, 2H), 3.83 (s, 3H), 3.70 (t , J=5.6Hz, 2H), 3.40 (q, J=7.2Hz, 2H), 2.43 (s, 3H), 1.02 (t, J=7.2Hz, 3H); 13CNMR (100MHz, DMSO-d6): δ156.75, 154.17.152.85, 148.39, 142.91, 136.73, 132.91, 128.33, 127.54, 127.15, 122.2 4, 122.16, 117.27, 111.37, 111.24, 110.03, 107.62, 94.53, 67.88, 65.62, 55.44, 43.41, 20.83, 14.97; HRMS (TOFMSES - ): m / zcalcdforC 24 H 23 ClN3O4(MH) - 452.1377, found 452.1372;mp 210℃

[0371] Example 10: Preparation of (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-fluoro-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbonylhydrazine (CAP-1049, compound 5)

[0372]

[0373] 1-PrOH (10 mL) and acetic acid (1-2 drops) were added to 5-methylbenzofuran-2-carbazide (66.6 mg, 0.35 mmol) and 2-chloro-1-(2-ethoxyethyl)-5-fluoro-1H-indole-3-carboxaldehyde (94.4 mg, 0.35 mmol), and the mixture was refluxed for 8 hours. The reaction solution was distilled under reduced pressure and dissolved in a small amount of CH2Cl2, then added dropwise to an n-Hex solution. 137 mg of the title compound was obtained by filtration of the resulting solid (89% yield, white solid).

[0374] 1 HNMR (400MHz, DMSO-d6): δ12.06 (s, 1H), 8.73 (s, 1H), 8.02 (dd, J=9.6, 2.8Hz, 1H), 7.67 (dd, J=9.2, 4.8Hz, 1H), 7.63-7.58 (m, 3H), 7.32 (d, J=9.6Hz, 1H), 7.18 (m, 1H), 4.47 (t, J=5.6Hz, 2H), 3.69 (t, J=5.6Hz, 2H), 3.38 (q, J=6.8Hz, 2H), 2.43 (s, 3H), 0.99 (t, J=6.8Hz, 3H); 13CNMR (100MHz, DMSO-d6): δ158.31 (d, J=233.8Hz), 154.27, 152.88, 148.31 , 142.49, 132.93, 132.47, 130.35, 128.37, 127.14, 123.65 (d, J=11.3Hz), 122.24, 112.30 (d, J=9.4Hz), 111.36, 111.21 (d, J=25.9Hz), 110.19, 107. 65 (d, J=4.4Hz), 106.37 (d, J=25.1Hz), 67.88, 65.64, 43.85, 20.82, 14.88; 19 FNMR (376MHz, DMSO-d6): δ-121.3 (s, 1F); HRMS (TOFMSES - ): m / zcalcdf orC 23 H 20 ClFN3O3(MH) - 440.1177, found440.1185;mp 187℃

[0375] Example 11: Preparation of (E)-N'-{[2,5-dichloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbonylhydrazine (CAP-1060, compound 9)

[0376]

[0377] 1-PrOH (30 mL) and acetic acid (1-2 drops) were added to 5-methylbenzofuran-2-carbazide (165.5 mg, 0.87 mmol) and 2,5-dichloro-1-(2-ethoxyethyl)-1H-indole-3-carboxaldehyde (249.0 mg, 0.87 mmol), and the mixture was refluxed for 3 hours. The reaction solution was distilled under reduced pressure and recrystallized with EtOH. The resulting solid was filtered and washed with cold EtOH to give 270 mg of the title compound (68% yield, white solid).

[0378] 1HNMR (400MHz, DMSO-d6): δ12.07 (s, 1H), 8.73 (s, 1H), 8.32 (s, 1H), 7.67 (d, J=8.8Hz, 1H), 7.63-7.58 (m, 3H), 7.35-7 .31 (m, 2H), 4.46 (t, J=5.0Hz, 2H), 3.69 (t, J=5.0Hz, 2H), 3.38 (q, J=7.0Hz, 2H), 2.43 (s, 3H), 0.98 (t, J=7.0Hz, 3H); 13 CNMR (100MHz, DMSO-d6): δ154.31, 152.92, 148.30, 142.52, 134.40, 132.98, 130.52, 128.44, 127.17, 126.42, 124.25, 1 23.16, 122.30, 120.44, 112.65, 111.42, 110.30, 107.33, 67.89, 65.68, 43.88, 20.88, 14.93; HRMS (ESI): m / zcalcdforC 23 H 22 Cl2N3O3(M+H) + 458.1038, found458.1037;mp 98℃

[0379] Example 12: Preparation of (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-(trifluoromethoxy)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbonylhydrazine (CAP-1047, compound 3)

[0380]

[0381] 1-PrOH (10 mL) and acetic acid (1-2 drops) were added to 5-methylbenzofuran-2-carbazide (22.8 mg, 0.12 mmol) and 2-chloro-1-(2-ethoxyethyl)-5-trifluoromethoxy-1H-indole-3-carboxaldehyde (40.3 mg, 0.12 mmol), and the mixture was refluxed for 18 hours. The reaction solution was subjected to vacuum distillation, and the residue was separated by column chromatography (n-Hex:EA = 1:2) to obtain 52 mg of the title compound (yield 85%, pale yellow solid).

[0382] 1HNMR (400MHz, DMSO-d6): δ12.10 (s, 1H), 8.75 (s, 1H), 8.26 (s, 1H), 7.45 (d, J=9.2Hz, 1H), 7.63-7.58 (m, 3H), 7.33-7 .29 (m, 2H), 4.49 (t, J = 5.2Hz, 2H), 3.71 (t, J = 5.2Hz, 2H), 3.39 (q, J = 7.2Hz, 2H), 2.43 (s, 3H), 0.99 (t, J = 7.2Hz, 3H); 13 CNMR (100MHz, DMSO-d6): δ154.27, 152.88, 148.29, 143.57, 142.30, 134.32, 132.95, 130.87, 128.41, 127.13, 123.45, 122.27, 120.34 (q, J=253.9Hz), 116.68, 113.47, 112.43, 111.37, 110.27, 107.92, 67.84, 65.63, 43.93, 20.83, 14.87; 19 FNMR (376MHz, DMSO-d6): δ-56.8 (s, 3F); HRMS (TOFMSES-): m / zcalcdforC 24 H 20 ClF3N3O4(MH)-506.1094, found506.1087; mp 84-85℃

[0383] Example 13: Preparation of (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-methyl-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbonylhydrazine (CAP-1059, Compound 8)

[0384]

[0385] 1-PrOH (30 mL) and acetic acid (1-2 drops) were added to 5-methylbenzofuran-2-carbonylhydrazine (374.7 mg, 1.97 mmol) and 2-chloro-1-(2-ethoxyethyl)-5-methyl-1H-indole-3-carboxaldehyde (523.5 mg, 1.97 mmol), and the mixture was heated under reflux for 3 hours. The reaction solution was distilled under reduced pressure and recrystallized with EtOH. The resulting solid was filtered and washed with cold EtOH to give 690 mg of the title compound (80% yield, pale yellow solid).

[0386] 1HNMR (400MHz, DMSO-d6): δ11.99 (s, 1H), 8.75 (s, 1H), 8.11 (s, 1H), 7.63-7.59 (m, 3H), 7.50 (d, J=8.4Hz, 1H), 7.33 (dd, J=8.4, 1.4Hz, 1H), 7.1 4 (dd, J=8.4, 1.4Hz, 1H), 4.43 (t, J=5.4Hz, 2H), 3.69 (t, J=5.4Hz, 2H), 3.39 (q, J=7.0Hz, 2H), 2.45 (s, 3H), 2.44 (s, 3H), 1.01 (t, J=7.0Hz, 3H); 13 CNMR (100MHz, DMSO-d6): δ154.21, 152.90, 148.46, 143.38, 134.16, 132.95, 130.58, 129.22, 128.37, 127.20, 124.64, 123.54 , 122.28, 121.08, 111.41, 110.42, 110.10, 107.09, 67.90, 65.66, 43.57, 21.34, 20.88, 14.95; HRMS (TOFMSES-): m / zcalcdforC 24 H 23 ClN3O3(MH) - 436.1428, found436.1442;mp 197℃

[0387] Example 14: Preparation of (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methoxybenzofuran-2-carbonylhydrazine (CAP-1051, compound 16)

[0388]

[0389] 1-PrOH (15 mL) and acetic acid (1-2 drops) were added to 5-methoxybenzofuran-2-carbonylhydrazine (103.1 mg, 0.50 mmol) and 2-chloro-1-(2-ethoxyethyl)-1H-indole-3-carboxaldehyde (125.9 mg, 0.50 mmol), and the mixture was refluxed for 7 hours. The reaction solution was distilled under reduced pressure, and the residue was washed with a 1:1 mixture of n-Hex / CH₂Cl₂ to give 204 mg of the title compound (93% yield, pale yellow solid).

[0390] 1HNMR (400MHz, DMSO-d6): δ12.01 (s, 1H), 8.75 (s, 1H), 8.32 (d, J=7.6Hz, 1H), 7.63-7.60 (m, 3H), 7.33-7.24 (m, 3H), 7.10 (dd , J=8.8, 2.4Hz, 1H), 4.46 (t, J=5.6Hz, 2H), 3.82 (s, 3H), 3.70 (t, J=5.6Hz, 2H), 3.39 (q, J=7.2Hz, 2H), 1.00 (t, J=7.2Hz, 3H); 13 CNMR (100MHz, DMSO-d6): δ156.11, 154.16, 149.34, 148.94, 143.01, 135.76, 129.27, 127.72, 123.35, 123.21, 121.68, 121 .43, 116.39, 112.47, 110.68, 110.43, 107.53, 104.20, 67.85, 65.63, 55.63, 43.53, 14.90; HRMS (TOFMSES-): m / zcalcdforC 23 H 21 ClN3O4(MH)-438.1221, found 438.1218; mp 185℃

[0391] Example 15: Preparation of (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl]methylene}-5-methoxybenzofuran-2-carbonylhydrazine (CAP-1053, compound 17)

[0392]

[0393] 1-PrOH (15 mL) and acetic acid (1-2 drops) were added to 5-methoxybenzofuran-2-carbonylhydrazine (103.1 mg, 0.50 mmol) and 2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indole-3-carboxaldehyde (140.9 mg, 0.50 mmol), and the mixture was refluxed for 8 hours. The reaction solution was subjected to vacuum distillation, and the residue was separated by column chromatography (n-Hex:EA = 2:3) to obtain 222 mg of the title compound (94% yield, pale yellow solid).

[0394] 1HNMR (400MHz, DMSO-d6): δ12.01 (s, 1H), 8.73 (s, 1H), 7.91 (d, J=2.4Hz, 1H), 7. 62-7.59 (m, 2H), 7.52 (d, J=8.8Hz, 1H), 7.32 (d, J=2.4Hz, 1H), 7.09 (dd, J=8.8, 2.4Hz, 1H), 6.95 (dd, J=8.8, 2.4Hz, 1H), 4.42 (t, J=5.2Hz, 2H), 3.82 (s, 3H), 3. 817 (s, 3H), 3.68 (t, J = 5.2Hz, 2H), 3.38 (q, J = 7.2Hz, 2H), 1.00 (t, J = 7.2Hz, 3H); 13 CNMR (100MHz, DMSO-d6): δ156.11, 155.21, 154.12, 149.33, 149.03, 142.97, 130.75, 128.96, 127.74, 124.02, 116.37, 112.45 , 112.27, 111.54, 110.42, 107.25, 104.20, 104.09, 67.92, 65.63, 55.63, 55.33, 43.67, 14.91; HRMS (TOFMSES-): m / zcalcdforC 24 H 23 ClN3O5(MH)-468.1326, found468.1322; mp 128-129℃

[0395] Example 16: Preparation of (E)-5-chloro-N'-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}benzofuran-2-carbonylhydrazine (CAP-1062, compound 20)

[0396]

[0397] 1-PrOH (15 mL) and acetic acid (1-2 drops) were added to 5-chlorobenzofuran-2-carbonylhydrazine (103.2 mg, 0.49 mmol) and 2-chloro-1-(2-ethoxyethyl)-1H-indole-3-carboxaldehyde (123.3 mg, 0.49 mmol), and the mixture was refluxed for 24 hours. The reaction solution was subjected to vacuum distillation, and the residue was separated by column chromatography (CH₂Cl₂:EA = 5:1) to obtain 188 mg of the title compound (yield 86%, pale yellow solid).

[0398] 1HNMR (400MHz, DMSO-d6): δ12.10 (s, 1H), 8.75 (s, 1H), 8.32 (d, J=7.6Hz, 1H), 7.94 (d, J=2.0Hz, 1H), 7.76 (d, J=9.2Hz, 1H), 7.69 (s, 1H), 7.61 (d, J=7. 6Hz, 1H), 7.53 (dd, J=9.2, 2.0Hz, 1H), 7.33-7.24 (m, 2H), 4.46 (t, J=5.2Hz , 2H), 3.70 (t, J=5.2Hz, 2H), 3.39 (q, J=7.2Hz, 2H), 1.00 (t, J=7.2Hz, 3H); 13 CNMR (100MHz, DMSO-d6): δ153.79, 152.86, 149.68, 143.29, 135.76, 129.43, 128.66, 128.15, 126.97, 123.33, 123.2 3, 122.21, 121.72, 121.39, 113.52, 110.71, 109.75, 107.44, 67.84, 65.63, 43.55, 14.90; HRMS (ESI): m / zcalcdforC 22 H 20 Cl2N3O3(M+H) + 444.0882, found 444.0884;mp 117℃

[0399] Example 17: Preparation of (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-4,7-dimethylbenzofuran-2-carbonylhydrazine (CAP-1063, compound 21)

[0400]

[0401] 1-PrOH (15 mL) and acetic acid (1-2 drops) were added to 4,7-dimethylbenzofuran-2-carbazide (81.7 mg, 0.40 mmol) and 2-chloro-1-(2-ethoxyethyl)-1H-indole-3-carboxaldehyde (100.7 mg, 0.40 mmol), and the mixture was refluxed for 24 hours. The reaction solution was subjected to vacuum distillation, and the residue was separated by column chromatography (CH₂Cl₂:EA = 5:1) to obtain 158 mg of the title compound (90% yield, white solid).

[0402] 1HNMR (400MHz, DMSO-d6): δ11.87 (s, 1H), 8.76 (s, 1H), 8.34 (d, J=7.2Hz, 1H), 7.76 (s, 1H), 7.61 (d, J=8.0Hz, 1H), 7.34-7.25 (m, 2H), 7.19 (d, J=8.0H z, 1H), 7.05 (d, J=7.2Hz, 1H), 4.46 (t, J=5.6Hz, 2H), 3.71 (t, J=5.6Hz, 2H ), 3.39 (q, J=7.2Hz, 2H), 2.54 (s, 3H), 2.51 (s, 3H), 1.00 (t, J=7.2Hz, 3H); 13 CNMR (100MHz, DMSO-d6): δ154.34, 153.35, 147.46, 142.86, 135.76, 129.60, 129.25, 127.53, 126.50, 123.81, 123.35, 123. 22, 121.67, 121.42, 118.71, 110.69, 109.48, 107.50, 67.85, 65.62, 43.53, 17.88, 14.90, 14.52; HRMS (ESI): m / zcalcdforC 24 H 25 ClN3O3(M+H) + 438.1584, found 438.1584; mp 202℃

[0403] Example 18: Preparation of (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-4,6-dimethoxybenzofuran-2-carbonylhydrazine (CAP-1064, compound 22)

[0404]

[0405] 1-PrOH (15 mL) and acetic acid (1 drop to 2 drops) were added to 4,6-dimethoxybenzofuran-2-carbonylhydrazine (118.1 mg, 0.50 mmol) and 2-chloro-1-(2-ethoxyethyl)-1H-indole-3-carboxaldehyde (125.9 mg, 0.50 mmol), and the mixture was refluxed for 4 hours. After cooling the reaction solution and filtering the solid, the solid was washed with EtOH to give 210 mg of the title compound (yield 89%, white solid).

[0406] 1HNMR (400MHz, DMSO-d6): δ11.82 (s, 1H), 8.69 (s, 1H), 8.32 (d, J=7.6Hz, 1H), 7.63 (s, 1H), 7.60 (d, J=7.6Hz, 1H), 7.33-7.23 (m, 2H), 6.87 (s, 1H) ), 6.50 (d, J = 2.0Hz, 1H), 4.45 (t, J = 5.6Hz, 2H), 3.92 (s, 3H), 3.85 (s, 3H), 3.70 (t, J = 5.6Hz, 2H), 3.38 (q, J = 7.2Hz, 2H), 1.00 (t, J = 7.2Hz, 3H); 13 CNMR (100MHz, DMSO-d6): δ161.07, 156.44, 154.11, 146.03, 142.31, 135.74, 129.04, 123.33, 123.18, 121.60, 121 .41, 110.98, 110.65, 107.58, 107.53, 95.13, 88.44, 67.85, 65.62, 55.84, 43.50, 14.90; HRMS (ESI): m / zcalcdforC 24 H 25 ClN3O5(M+H) + 470.1483, found 470.1482; mp 215℃

[0407] Example 19: Preparation of (E)-N'-{2-[2-((2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl)methylene]hydrazine-1-carbonyl}benzofuran-5-yl)acetamide (CAP-1056, compound 18)

[0408]

[0409] 1-PrOH (20 mL) and acetic acid (1 drop to 2 drops) were added to N-(2-(hydrazine carbonyl)benzofuran-5-yl)acetamide (116.6 mg, 0.50 mmol) and 2-chloro-1-(2-ethoxyethyl)-1H-indole-3-carboxaldehyde (125.9 mg, 0.50 mmol), and the mixture was refluxed for 12 hours. After cooling the reaction solution and filtering the solid, the solid was washed with EtOH to give 166 mg of the title compound (71% yield, pale yellow solid).

[0410] 1HNMR (400MHz, DMSO-d6): δ12.02(s, 1H), 10.07(s, 1H), 8.76(s, 1H), 8.32( d, J=7.2Hz, 1H), 8.17 (d, J=2.4Hz, 1H), 7.68 (s, 1H), 7.65-7.60 (m, 2H), 7. 53 (dd, J=9.2, 2.4Hz, 1H), 7.33-7.24 (m, 2H), 4.46 (t, J=5.6Hz, 2H), 3.70 ( t, J=5.6Hz, 2H), 3.39 (q, J=7.2Hz, 2H), 2.08 (s, 3H), 1.00 (t, J=7.2Hz, 3H); 13 CNMR (100MHz, DMSO-d6): δ168.24, 154.15, 150.62, 148.84, 143.08, 135.77, 135.54, 129.32, 127.18, 123.36, 123.24, 121.70, 121.44, 119.61, 112.10, 111.83, 110.72, 110.66, 107.53, 67.88, 65.65, 43.55, 23.97, 14.94; HRMS (TOFMSES-): m / zcalcdforC 24 H 22 ClN4O4(MH)-465.1330, found 465.1323; mp226℃

[0411] Example 20: Preparation of (E)-ethyl-2-(3-((2-(4,6-dimethoxybenzofuran-2-carbonyl)hydrazine)methyl)-2-methyl-1H-indol-1-yl)acetate (CAP-1061, compound 23)

[0412]

[0413] 1-PrOH (15 mL) and acetic acid (1 drop to 2 drops) were added to 4,6-dimethoxybenzofuran-2-carbonylhydrazine (118.1 mg, 0.50 mmol) and ethyl 2-(3-formyl-2-methyl-1H-indol-1-yl)acetate (122.6 mg, 0.50 mmol), and the mixture was refluxed for 4 hours. After cooling the reaction solution and filtering the solid, the solid was washed with EtOH to give 197 mg of the title compound (yield 85%, white solid).

[0414] HRMS(ESI): m / zcalcdforC25H25N3O6Na(M+Na)486.1641, foun d 486.1642

[0415] Example 21: Preparation of (E)-2-(2-methyl-3-((2-(5-methylbenzofuran-2-carbonyl)hydrazine)methyl)-1H-indol-1-yl)ethyl acetate (CAP-1034, compound 10)

[0416]

[0417] 1-PrOH (15 mL) and acetic acid (1-2 drops) were added to 5-methylbenzofuran-2-carbonylhydrazine (82.5 mg, 0.43 mmol) and ethyl 2-(3-formyl-2-methyl-1H-indol-1-yl)acetate (105.5 mg, 0.43 mmol), and the mixture was refluxed for 36 hours. After distilling the reaction solution under reduced pressure, the residue was washed with Et2O to obtain 176 mg of the target compound (98%, white solid).

[0418] 1 HNMR (400MHz, DMSO-d6) δ11.77(s,1H),8.83(s,1H),8.30(m,1H),7.61-7.58(m,3H),7.48(m,1H),7.32(d,J=8.8 Hz,1H),7.22-7.19(m,2H),5.19(s,2H),4.19(q,J=7.2Hz,2H),2.49(s,3H),2.44(s,3H),1.23(t,J=7.2Hz,3H); 13 CNMR(100MHz,D MSO-d6)δ168.64,153.98,152.84,148.71,145.25,141.50,137.05,132.92,128.26,127.25,124.69,122.27,122.23 ,121.40,121.02,111.37,109.86,109.57,108.23,61.24,44.44,20.87,14.05,9.98; HRMS(TOFMSES-):m / zcalcdforC 24 H 22 N3O4(MH) - 416.1610,found 416.1628;mp186℃

[0419] Example 22: Preparation of (E)-2-(3-((2-(5-chlorobenzofuran-2-carbonyl)hydrazine)methyl)-2-methyl-1H-indole-1-yl-ethyl acetate (CAP-1035, compound 11)

[0420]

[0421] 1-PrOH (15 mL) and acetic acid (1 drop to 2 drops) were added to 5-benzofuran-2-carbonylhydrazine (105.3 mg, 0.50 mmol) and ethyl 2-(3-formyl-2-methyl-1H-indol-1-yl)acetate (122.6 mg, 0.50 mmol), and the mixture was refluxed for 24 hours. After cooling the reaction solution and filtering the solid, the solid was washed with EtOH to obtain 107 mg of the target compound (49%, pale yellow solid).

[0422] 1 HNMR (400MHz, DMSO-d6) δ11.84(s,1H),8.81(s,1H),8.28(m,1H),7.93(d,J=2.0Hz,1H),7.76(d,J=8.8Hz,1H),7.66(s,1H),7.5 2(dd,J=8.8,2.0Hz,1H),7.47(m,1H),7.22-7.17(m,2H),5.18(s,2H),4.18(q,J=6.8Hz,2H),2.49(s,3H),1.23(t,J=6.8Hz,3H); 13 CNMR(100MHz,DMSO-d6)δ168.63,153.56,152.84,150.01,145.56,141.69,137.05,128.76,128.16,126.89,124 .67,122.31,122.17,121.39,121.07,113.51,109.59,109.55,108.17,61.25,44.45,14.06,9.99; HRMS(TOFMSES - ):m / zcalcdforC 23 H 19 ClN3O4(MH) - 436.1064,found 436.1062;mp 214℃

[0423] Example 23: Preparation of (E)-2-(2-chloro-3-((2-(5-methylbenzofuran-2-carbonyl)hydrazine)methyl)-1H-indol-1-yl)acetate (CAP-1048, compound 4)

[0424]

[0425] 5-Methylbenzofuran-2-carbonylhydrazine (319.5 mg, 1.68 mmol) was reacted with methyl 2-(2-chloro-3-formyl-1H-indol-1-yl)acetate (422.8 mg, 1.68 mmol) by adding 20 mL of 1-PrOH and 1-2 drops of acetic acid and refluxing for 15 hours. After cooling the reaction solution and filtering the solid, the solid was washed with n-hexane to give 648 mg of the target compound (91%, but a yellow solid).

[0426] 1 HNMR(400MHz,DMSO-d6)δ12.01(s,1H),8.75(s,1H),8.33(d,J=7.5Hz,1H),7 .63-7.58(m,4H),7.33-7.24(m,3H),4.48(s,2H),3.68(s,3H),2.43(s,3H); 13 CNMR(100MHz,DMSO-d6)δ166.12,154.23,152.88,148.40,142.97,135.76,132.93,129.20,128.36,127.1 6,123.34,123.25,122.25,121.68,121.44,111.38,110.68,110.10,107.55,51.60,20.84; HRMS(TOFMSES - ):m / zcalcdforC 22 H 17 ClN3O4(MH) - 422.0908,found 422.0902;mp 212℃

[0427] The compounds prepared in the examples described above were subjected to the following experiments.

[0428] Experimental Example 1: Confirmation of the effect of inhibiting the nuclear factor κB inflammatory response pathway through cell-based reporter gene analysis

[0429] After transfecting cultured human-derived HEK293 cell lines with a nuclear factor κB luciferase (reporter) vector, treatment with 10 ng / ml TNF-α induced activation of the nuclear factor κB signaling pathway. Furthermore, during this process, treatment with 1 μM of each of nine compounds of Formula 1 was performed to determine their effects on inhibiting nuclear factor κB-mediated inflammatory response pathways. Specifically, the relative expression level of nuclear factor κB was confirmed.

[0430] like Figure 1As shown, the positive control group treated only with TNF-α showed a significant increase in reporter gene activity due to activation of the nuclear factor κB inflammatory response pathway. Conversely, the activity of the reporter gene was significantly increased in the group treated with compound of formula 1. Figure 1 When the compounds are represented as compounds 1 to 9 respectively, it was observed that the activity of nuclear factor κB was inhibited by more than 50%.

[0431] Experiment Example 2: Confirming the effect of inhibiting the phosphorylation of p65 (RelA), a key factor in the nuclear factor κB inflammatory response pathway.

[0432] Existing research on the regulation of κB inflammatory responses indicates that the phosphorylation of p65(RelA) is crucial for inducing inflammatory responses within the pathway. This is because phosphorylated p65 essentially migrates into the nucleus of immune cells to increase the production of cytokines that trigger inflammatory responses.

[0433] After culturing the same HEK293 cell line, each compound was treated to confirm whether it inhibited phosphorylation of p65(RelA), which is the most important process in nuclear factor κB signaling.

[0434] like Figure 2 As shown, the compound of chemical formula 1 of the present invention has been confirmed (in... Figure 2 All compounds (referred to as compounds 1 to 9) inhibit the phosphorylation of p65.

[0435] As described above, it has been confirmed that the compounds of the present invention effectively inhibit the nuclear factor κB inflammatory response pathway and inhibit the phosphorylation of p65 (RelA), an important factor in the nuclear factor κB signaling process, thereby confirming their excellent anti-inflammatory efficacy.

[0436] Experiment Example 3: Confirmation of the effect of inhibiting the nuclear factor κB inflammatory response pathway through cell-based reporter gene analysis.

[0437] HEK293 cells, a human-derived embryonic kidney cell line, were cultured, transfected with a luciferase vector for nuclear factor κB, and treated with 1 μM of the compound, followed by treatment with 10 ng / ml TNF-α to induce the nuclear factor κB pathway. The effect on nuclear factor κB gene expression was quantitatively measured using a luciferase reporter assay with a luminometer.

[0438] The results are as follows Figure 3 As shown.

[0439] pass Figure 3 It was confirmed that nuclear factor κB gene expression was high in the positive control group treated only with TNF-α. However, when the compounds of the embodiments of the present invention were treated (indicated by CAP-No. (+“No”)), inhibition of nuclear factor κB inflammatory signaling was observed.

[0440] In particular, in samples treated with the five compounds of Example 4 (CAP-1042), Example 12 (CAP-1047), Example 10 (CAP-1049), Example 13 (CAP-1059), and Example 11 (CAP-1060), the expression level of the nuclear factor κB gene was significantly reduced.

[0441] Experiment 4: Confirming the anti-inflammatory effect in the skin

[0442] To induce an immune response, 150 μl of 3% oxazolone was applied to the abdominal skin of 6-week-old mice. An inflammatory response was then induced by applying 20 μl of 0.5% oxazolone to the ears of the mice five times at daily intervals. Thirty minutes after oxazolone application, 20 μl of either Example 4 (CAP-1042) or Example 12 (CAP-1047) at a concentration of 500 nM was applied five times.

[0443] The number of mice in each experimental group was kept at more than 10, and the experiment was repeated twice.

[0444] 1. Confirm the anti-inflammatory effect with the naked eye.

[0445] As a result of the above experiments, the anti-inflammatory effect confirmed by the naked eye and the anti-inflammatory results confirmed by hematoxylin and eosin staining are as follows: Figure 4 and Figure 5 As shown.

[0446] Specifically, Figure 4 The results show the inflammatory changes confirmed by visual observation. Mice treated with oxazolone alone showed redness in their ears compared to those treated with ethanol as a solvent, indicating a well-induced inflammatory response. Conversely, in the animal groups treated with compound (1042) of Example 4 or (CAP-1047) of Example 12, skin conditions very similar to those observed with ethanol treatment were observed.

[0447] Furthermore, the results of hematoxylin-eosin staining are as follows: Figure 5As shown. When skin inflammation is induced, a pathological phenomenon known as epidermal hyperplasia, characterized by an increase in both the overall thickness of the skin and the thickness of the epithelium, is typically observed. Compared to skin treated with ethanol as a solvent alone, the overall thickness of skin treated with oxazolone alone increased by approximately two times, and the thickness of the epithelium also increased by approximately three times. Conversely, when treated with the compound of Example 4 (CAP-1042) or Example 12 (CAP-1047), the overall thickness of the skin and the thickness of the epithelium were reduced to a level very similar to the normal condition after ethanol treatment, confirming the excellent anti-inflammatory efficacy of the compound. In particular, a superior effect compared to the positive control group was observed in its effect of inhibiting epidermal hyperplasia.

[0448] 2. Confirmed reduction in the number of inflammatory immune cells

[0449] If skin inflammation is induced, a pathological phenomenon of increased number of inflammatory immune cells in the skin as a whole is usually observed.

[0450] Therefore, in this invention, the therapeutic effect of the compound of this invention is confirmed by measuring changes in the number of inflammatory immune cells.

[0451] Specifically, in hematoxylin and eosin staining ( Figure 5 Then, a microscope was used to take photographs evenly at more than 10 locations on each sample, and the average value was obtained by directly counting the number of inflammatory immune cells stained dark red in the photographs.

[0452] The results are as follows Figure 6 As shown.

[0453] pass Figure 6 It was confirmed that the number of inflammatory immune cells in skin treated with oxazolone alone increased by approximately three times compared to skin treated with ethanol as a solvent alone. Conversely, the number of immune cells treated with the compounds of Example 4 (1042) or Example 7 (1047) of this application decreased to a level very similar to that of normal skin treated with ethanol.

[0454] 3. Confirm the reduction of inflammatory cytokines

[0455] After the above-described application experiment demonstrating the nuclear factor κB signaling process, skin tissue was collected from each mouse, and ribonucleic acid (RNA) was extracted after pulverizing the tissue using a tissue homogenizer. The extracted RNA was then reverse transcribed using complementary deoxyribonucleic acid (cDNA), and the levels of cytokines IL-4 and IL-13 present in the skin tissue were measured using real-time polymerase chain reaction (Real-time PCR) followed by quantitative polymerase chain reaction (qPCR).

[0456] The results are as follows Figure 7 As shown.

[0457] pass Figure 7 It was confirmed that the levels of interleukin-4 (IL-4) and interleukin-13 (IL-13), which are inflammatory cytokines, increased in the ear tissues of mice treated with oxazolone alone to induce inflammation. However, in mice treated with oxazolone in conjunction with the compounds of Example 4 (1042) or Example 7 (1047) of the present invention, the expression levels of the above cytokines were confirmed to be significantly reduced to 20%-25%.

[0458] The above results confirm that the fabric of the present invention exhibits excellent anti-inflammatory effects.

[0459] Experiment Example 5: Confirmation of the inhibitory effect of nuclear factor κB on typical and atypical inflammatory signal transduction activities and the reduction of target gene expression.

[0460] Nuclear factor κB signaling generally involves both canonical and atypical pathways. In the canonical pathway, when an inflammatory response is induced by treatment of TNF-α, the amount of p65 protein phosphorylated to P-p65 protein increases, inducing an inflammatory response as it moves into the nucleus. In the atypical pathway, when an inflammatory response is induced by LIGHT compounds that activate the non-canonical NF-κB pathway, p100 protein is converted to p52 protein through a treatment process and moves into the nucleus to induce an inflammatory response.

[0461] For these different pathways, it was determined whether the compounds of the present invention exhibited pathway inhibitory effects. Specifically, to determine whether they modulated the activity of nuclear factor κB signaling (typical and atypical pathways), Western blotting was used to confirm the regulation of phosphorylated p65 and p100 proteins, which are major signaling regulatory molecules. In this experiment, the compound of Example 4 (1042) was used as the compound of the present invention.

[0462] The results are as follows Figure 8 As shown.

[0463] exist Figure 8 It can be confirmed that in the typical pathway, if TNFα treatment induces an inflammatory response, the inflammatory response is induced (+) as the amount of P-p65 protein, the phosphorylated product of p65 protein, increases and it migrates into the nucleus. Furthermore, if an inflammatory response is induced by the LIGHT compound, the inflammatory response is induced (+) as p100 protein is converted to p52 protein through treatment and migrates into the nucleus. TPCA1, known to act on p65 protein, functions in the typical pathway but not in the atypical pathway.

[0464] Conversely, the compounds of the present invention act as dual inhibitors in both typical and atypical pathways of nuclear factor κB inflammatory signal transduction, exhibiting excellent pathway inhibition effects in both pathways.

[0465] The above results confirm that the compounds of the present invention exhibit a dual inhibitory effect on both typical and atypical pathways of nuclear factor κB inflammatory signal transduction.

[0466] Experimental Example 6: Confirmation of the selective binding of the compound to IKKα and its inhibition of structure formation.

[0467] To confirm the regulatory mechanism of nuclear factor κB activity, biotin was incorporated into the drug (Example 4) to investigate whether it binds to the major regulatory molecules of nuclear factor κB. Specifically, it was determined whether it binds to IKKα, IKKβ, IKKγ, p65, and IκBα in the relevant canonical and atypical pathways.

[0468] The results are as follows Figure 9 As shown.

[0469] pass Figure 9 It can be confirmed that it specifically and selectively binds to IKKα in the major regulatory molecule of nuclear factor κB.

[0470] In the case of IKKα, the nuclear factor κB signaling process is mediated in the form of a hexamer (6mer), that is, a complex of 6 monomers.

[0471] Therefore, the active form in the form of a hexamer was confirmed by treating the compound of the present invention (Example 4), and the results were as follows: Figure 10 As shown.

[0472] pass Figure 10 It can be confirmed that the compound used in the embodiments of the present invention inhibits the formation of hexamer IKKα, thereby acting as a selective inhibitor of IKKα.

[0473] Experiment Example 7: Confirming the efficacy of inhibiting cytokine storm and treating sepsis using a sepsis model

[0474] Since there are no disease model animals that die from coronaviruses, especially from cytokine storms caused by severe acute respiratory syndrome coronavirus 2, this invention uses an acute sepsis model that induces cytokine storms due to an inflammatory response to confirm the possibility of inhibiting cytokine storms, treating severe acute respiratory syndrome coronavirus 2 infection, and treating sepsis.

[0475] The experiment utilized an animal model of sepsis (death) induced by lipopolysaccharide (LPS) treatment (a cytokine storm-like model). The experimental design was as follows: Figure 11a As shown. After induced sepsis by intraperitoneal administration of lipopolysaccharide and intravenous injection of the drug (Example 4), blood samples were taken at 3 hours, 6 hours, and 12 hours, respectively. Survival rate was confirmed within 48 hours to observe sepsis-induced mortality.

[0476] The results of confirming the survival rate after 48 hours are as follows: Figure 11b As shown, it can be confirmed that administration of the compound of Example 4 of the present invention significantly reduces the mortality rate caused by sepsis.

[0477] And, as Figure 11c As shown, it can be confirmed that it has excellent effects in inhibiting the production of cytokines and chemokines.

[0478] The above results confirm that the compounds in the embodiments of the present invention reduce the mortality rate caused by sepsis by inhibiting the expression of cytokines and chemokines in vivo, and confirm that they can inhibit acute inflammatory responses, especially cytokine storms.

Claims

1. Use of compounds selected from the following, their stereoisomers, or pharmaceutically acceptable salts thereof in the preparation of medicaments for the treatment of dermatitis. (E)-2-{2-chloro-3-[(2-(5-methylbenzofuran-2-carbonyl)hydrazine)methyl]-1H-indol-1-yl}ethyl acetate; (E)-N'-{[1-(2-ethoxyethyl)-2-(trifluoromethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbazide; (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbazide; (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-6-methoxy-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbazide; (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-fluoro-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbazide; (E)-N'-{[2,5-dichloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbazide; (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-(trifluoromethoxy)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbazide; (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-methyl-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbazide; and (E)-2-(2-chloro-3-((2-(5-methylbenzofuran-2-carbonyl)hydrazine)methyl)-1H-indol-1-yl)acetate, The dermatitis mentioned is selected from any one of atopic dermatitis, contact dermatitis, allergic dermatitis, and pruritus.

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