Novel compound and composition comprising same as active ingredient for inhibiting hepatitis b virus (HBV) proliferation

Abstract

The present application relates to a compound represented by chemical formula 1: [Chemical formula 1] (In chemical formula 1, R is a substitutable linear or branched alkyl group of C1-C20, a substitutable cycloalkyl group of C3-C20, or a substitutable aryl group of C6-C20, wherein the substitution is performed with oxygen, nitrogen, sulfur, hydroxy, a linear or branched alkyl group of C1-C6, a cycloalkyl group of C3-C20, an aryl group of C6-C20, a halogen, or a combination thereof).

Description

Novel compound and composition for inhibiting hepatitis B virus (HBV) replication containing the same as an active ingredient

[0001] The present invention relates to a novel compound and a composition for inhibiting the proliferation of hepatitis B virus (HBV) containing the same as an active ingredient.

[0002] Hepatitis B virus (HBV) is one of the major pathogens causing serious health problems worldwide. According to the World Health Organization (WHO), approximately 250 million people are infected with chronic hepatitis B, and about 900,000 people die each year from hepatitis B-related complications.

[0003] Currently, nucleoside / nucleotide analogs and interferon-alpha are primarily used to treat hepatitis B. However, these treatments have drawbacks, such as difficulty in achieving complete viral elimination, the development of resistance with long-term administration, and side effects. Therefore, there is an urgent need for the development of anti-HBV drugs with novel mechanisms of action.

[0004] Recent studies have revealed that the sirtuin protein family plays a crucial role in the HBV life cycle. In particular, Sirtuin 2 (SIRT2) is known to act as an important regulator in the process of HBV replication. Consequently, interest in developing inhibitors targeting SIRT2 is growing. AGK2 is one of the well-known SIRT2 inhibitors that demonstrated an inhibitory effect on HBV proliferation in in vitro experiments; however, its clinical application has been limited due to low selectivity and side effects.

[0005] Therefore, there is a need for research to improve the efficacy of inhibiting HBV proliferation and reduce side effects through the development of new variants based on the structure of AGK2.

[0006] Korean Patent Publication No. 10-2020-0123392, which is the background technology of the present invention, relates to a composition for inhibiting the proliferation of hepatitis B virus and a method thereof.

[0007] The present invention aims to solve the problems of the aforementioned prior art by providing a compound represented by the following chemical formula 1, which is a new variant based on the structure of AGK2, and a method for preparing the same:

[0008] [Chemical Formula 1]

[0009]

[0010] (In the above chemical formula 1,

[0011] R is a linear or branched C1-C that can be substituted. 20 alkyl groups of, substitutable C3-C 20 cycloalkyl groups or C6-C that can be substituted 20 Arilgi,

[0012] The above substitutions are oxygen, nitrogen, sulfur, hydroxyl, linear or branched C1-C6 alkyl groups, C3-C 20 cycloalkyl group of, C6-C 20 (Substituted by an aryl group, a halogen, or a combination thereof).

[0013] In addition, a composition for inhibiting HBV proliferation is provided, comprising the above compound as an active ingredient.

[0014] In addition, a pharmaceutical composition for preventing or treating HBV infection is provided, comprising the above-mentioned composition for inhibiting HBV proliferation.

[0015] However, the technical problems that the embodiments of the present invention aim to solve are not limited to the technical problems described above, and other technical problems may exist.

[0016] As a technical means for achieving the above-mentioned technical problem, the first aspect of the present invention provides a compound represented by the following chemical formula 1:

[0017] [Chemical Formula 1]

[0018]

[0019] (In the above chemical formula 1,

[0020] R is a linear or branched C1-C that can be substituted. 20 alkyl groups of, substitutable C3-C 20 cycloalkyl groups or C6-C that can be substituted 20 Arilgi,

[0021] The above substitutions are oxygen, nitrogen, sulfur, hydroxyl, linear or branched C1-C6 alkyl groups, C3-C 20 cycloalkyl group of, C6-C 20 (Substituted by an aryl group, a halogen, or a combination thereof).

[0022] According to one embodiment of the present invention, the compound represented by Formula 1 may include any one of the following compounds, but is not limited thereto:

[0023] ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; .

[0024] Additionally, the second aspect of the present invention provides a method for preparing a compound according to the first aspect of the present invention, comprising the steps of: reacting a compound represented by the following chemical formula 2 and a compound represented by the following chemical formula 3 to prepare a compound represented by the following chemical formula 4; reacting the compound represented by the following chemical formula 4 with a compound represented by the following chemical formula 5 to prepare a compound represented by the following chemical formula 6; and substituting a hydroxyl group of the compound represented by the following chemical formula 6 to prepare a compound represented by the chemical formula 1 of the first aspect of the present invention.

[0025] [Chemical Formula 2]

[0026]

[0027] (In the above chemical formula 2, X 1 (is a halogen or B(OH)2)

[0028] [Chemical Formula 3]

[0029]

[0030] (In the above chemical formula 3, X 2 is a halogen or B(OH)2)

[0031] [Chemical Formula 4]

[0032]

[0033] [Chemical Formula 5]

[0034]

[0035] [Chemical Formula 6]

[0036] .

[0037] According to one embodiment of the present invention, the above X 1 and X 2 One of them may be a halogen and the other may be B(OH)2, but is not limited thereto.

[0038] In addition, the third aspect of the present invention provides a composition for inhibiting HBV proliferation, comprising a compound according to the first aspect of the present invention as an active ingredient.

[0039] According to one embodiment of the present invention, HBc (Hepatitis B core) protein expression in HepG2 cells may be inhibited by a compound represented by the above chemical formula 1, but is not limited thereto.

[0040] According to one embodiment of the present invention, the formation of nucleocapsid in HepG2 cells may be inhibited by the compound, but is not limited thereto.

[0041] According to one embodiment of the present invention, HBV DNA synthesis in HepG2 cells may be inhibited by the compound, but is not limited thereto.

[0042] According to one embodiment of the present invention, HBV DNA synthesis in Huh7 cells may be inhibited by the compound, but is not limited thereto.

[0043] In addition, the fourth aspect of the present invention provides a pharmaceutical composition for preventing or treating HBV infection, comprising a composition for inhibiting HBV proliferation according to the third aspect of the present invention.

[0044] According to one embodiment of the present invention, the pharmaceutical composition for preventing or treating HBV infection may additionally comprise a pharmaceutically acceptable carrier, but is not limited thereto.

[0045] According to one embodiment of the present invention, the carrier may comprise, but is not limited to, a substance selected from the group consisting of lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, menthol, mineral oil, and combinations thereof.

[0046] According to one embodiment of the present invention, the pharmaceutical composition for preventing or treating HBV infection may additionally include a component selected from the group consisting of preservatives, solubilizers, stabilizers, humectants, sweeteners, coloring agents, flavoring agents, salts, buffers, antioxidants, lubricants, emulsifiers, suspending agents, preservatives, and combinations thereof, but is not limited thereto.

[0047] According to one embodiment of the present invention, the pharmaceutical composition for preventing or treating HBV infection may be administered by a method selected from the group consisting of intraperitoneal administration, oral administration, inhalation administration, intravenous administration, intramuscular administration, subcutaneous administration, dermal administration, intrauterine administration, tumor administration, rectal administration, and combinations thereof, but is not limited thereto.

[0048] The means for solving the problem described above are merely exemplary and should not be interpreted as intended to limit the present invention. In addition to the exemplary embodiments described above, additional embodiments may exist in the drawings and the detailed description of the invention.

[0049] The novel compound according to the present invention is a new variant based on the AGK2 structure, and as a result of the MTS assay targeting HepG2 liver cancer cell lines, it did not exhibit cytotoxicity in various concentration ranges and showed high safety by exhibiting toxicity so low that the CC50 value (50% cytotoxicity concentration) could not be calculated.

[0050] In addition, it was confirmed that it has superior efficacy in inhibiting HBV proliferation compared to AGK2, and effectively inhibits the proliferation of HBV in HepG2 cells. Specifically, it showed effects of inhibiting HBc(C) protein expression, inhibiting core particle (nucleocapsid or capsid) formation, and inhibiting HBV DNA synthesis.

[0051] In addition, it was confirmed to inhibit HBV DNA synthesis in Huh7 cells as well as HepG2 cells, indicating that it can be effective in various hepatocyte environments.

[0052] In addition, viral replication can be effectively blocked by simultaneously inhibiting various stages of the HBV life cycle (protein expression, particle formation, DNA synthesis).

[0053] However, the effects obtainable from this invention are not limited to those described above, and other effects may exist.

[0054] Figure 1 is the NMR spectrum of a compound according to Example 1 of the present invention.

[0055] Figure 2 is the NMR spectrum of a compound according to Example 2 of the present invention.

[0056] Figure 3 is the NMR spectrum of a compound according to Example 3 of the present invention.

[0057] Figure 4 is the NMR spectrum of a compound according to Example 4 of the present invention.

[0058] Figure 5 is the NMR spectrum of a compound according to Example 5 of the present invention.

[0059] Figure 6 is the NMR spectrum of a compound according to Example 6 of the present invention.

[0060] Figure 7 is the NMR spectrum of a compound according to Example 7 of the present invention.

[0061] Figure 8 is the NMR spectrum of a compound according to Example 8 of the present invention.

[0062] Figure 9 is the NMR spectrum of a compound according to Example 9 of the present invention.

[0063] Figure 10 is the NMR spectrum of a compound according to Example 10 of the present invention.

[0064] Figure 11 is the NMR spectrum of a compound according to Example 11 of the present invention.

[0065] Figure 12 is the NMR spectrum of a compound according to Example 12 of the present invention.

[0066] Figure 13 is the NMR spectrum of a compound according to Example 13 of the present invention.

[0067] Figure 14 is the NMR spectrum of a compound according to Example 14 of the present invention.

[0068] Figure 15 is the NMR spectrum of a compound according to Example 15 of the present invention.

[0069] Figure 16 is the NMR spectrum of a compound according to Example 16 of the present invention.

[0070] Figure 17 is the NMR spectrum of a compound according to Example 17 of the present invention.

[0071] Figure 18 is the NMR spectrum of a compound according to Example 18 of the present invention.

[0072] Figure 19 is the NMR spectrum of a compound according to Example 19 of the present invention.

[0073] FIG. 20 is the NMR spectrum of a compound according to Example 20 of the present invention.

[0074] Figure 21 is the NMR spectrum of a compound according to Example 21 of the present invention.

[0075] Figure 22 is the NMR spectrum of a compound according to Example 22 of the present invention.

[0076] FIG. 23 is the NMR spectrum of a compound according to Example 23 of the present invention.

[0077] Figure 24 is the NMR spectrum of a compound according to Example 24 of the present invention.

[0078] Figures 25a and 25b are the results of a cytotoxicity test according to one experimental example of the present invention.

[0079] Figure 26 is the result of an experiment on the inhibition of HBV proliferation in liver cancer cell lines by a compound according to Example 1 of the present invention.

[0080] Embodiments of the present invention are described below with reference to the attached drawings to enable those skilled in the art to easily implement the invention. However, the present invention may be embodied in various different forms and is not limited to the embodiments described herein. Furthermore, in order to clearly explain the present invention in the drawings, parts unrelated to the explanation have been omitted, and similar parts throughout the specification are denoted by similar reference numerals.

[0081] Throughout this specification, when a part is described as being "connected" to another part, this includes not only cases where they are "directly connected," but also cases where they are "electrically connected" with other elements interposed between them.

[0082] Throughout the entire specification, when a component is described as being located "on," "on top," "on top," "under," "on bottom," or "on bottom" of another component, this includes not only cases where the component is in contact with the other component but also cases where another component exists between the two components.

[0083] Throughout this specification, when a part is described as "comprising" a certain component, this means that, unless specifically stated otherwise, it does not exclude other components but may include additional components.

[0084] As used herein, terms of degree such as “about,” “substantially,” etc., are used to mean at or near the stated value when inherent manufacturing and material tolerances are presented in the stated meaning, and are used to prevent unscrupulous infringers from unfairly exploiting the disclosure in which precise or absolute values ​​are mentioned to aid in understanding the invention. Furthermore, throughout this specification, “a step of” or “a step of” does not mean “a step for”.

[0085] Throughout this specification, the term “combination thereof” included in the Markush-type expression means one or more mixtures or combinations selected from the group consisting of the components described in the Markush-type expression, and means including one or more selected from the group consisting of said components.

[0086] Throughout the entire specification, the description "A and / or B" means "A, B, or A and B".

[0087] Hereinafter, the novel compounds of the present invention and compositions for inhibiting HBV proliferation containing the same as active ingredients will be described in detail with reference to embodiments, examples, and drawings. However, the present invention is not limited to these embodiments, examples, and drawings.

[0088]

[0089] As a technical means for achieving the above-mentioned technical problem, the first aspect of the present invention provides a compound represented by the following chemical formula 1:

[0090] [Chemical Formula 1]

[0091]

[0092] (In the above chemical formula 1,

[0093] R is a linear or branched C1-C that can be substituted. 20 alkyl groups of, substitutable C3-C 20cycloalkyl groups or C6-C that can be substituted 20 Arilgi,

[0094] The above substitutions are oxygen, nitrogen, sulfur, hydroxyl, linear or branched C1-C6 alkyl groups, C3-C 20 cycloalkyl group of, C6-C 20 (Substituted by an aryl group, a halogen, or a combination thereof).

[0095] The novel compound according to the present invention is a new variant based on the AGK2 structure, and as a result of the MTS assay targeting HepG2 liver cancer cell lines, it did not exhibit cytotoxicity in various concentration ranges and showed high safety by exhibiting toxicity so low that the CC50 value (50% cytotoxicity concentration) could not be calculated.

[0096] In addition, it was confirmed that it has superior efficacy in inhibiting HBV proliferation compared to AGK2, and effectively inhibits the proliferation of HBV in HepG2 cells. Specifically, it showed effects of inhibiting HBc(C) protein expression, inhibiting core particle (nucleocapsid or capsid) formation, and inhibiting HBV DNA synthesis.

[0097] In addition, it was confirmed to inhibit HBV DNA synthesis in Huh7 cells as well as HepG2 cells, indicating that it can be effective in various hepatocyte environments.

[0098] In addition, viral replication can be effectively blocked by simultaneously inhibiting various stages of the HBV life cycle (protein expression, particle formation, DNA synthesis).

[0099] According to one embodiment of the present invention, the compound represented by Formula 1 may include any one of the following compounds, but is not limited thereto:

[0100] ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; .

[0101] Additionally, the second aspect of the present invention provides a method for preparing a compound according to the first aspect of the present invention, comprising the steps of: reacting a compound represented by the following chemical formula 2 and a compound represented by the following chemical formula 3 to prepare a compound represented by the following chemical formula 4; reacting the compound represented by the following chemical formula 4 with a compound represented by the following chemical formula 5 to prepare a compound represented by the following chemical formula 6; and substituting a hydroxyl group of the compound represented by the following chemical formula 6 to prepare a compound represented by the chemical formula 1 of the first aspect of the present invention.

[0102] [Chemical Formula 2]

[0103]

[0104] (In the above chemical formula 2, X 1 (is a halogen or B(OH)2)

[0105] [Chemical Formula 3]

[0106]

[0107] (In the above chemical formula 3, X 2 is a halogen or B(OH)2)

[0108] [Chemical Formula 4]

[0109]

[0110] [Chemical Formula 5]

[0111]

[0112] [Chemical Formula 6]

[0113] .

[0114] Regarding the method for manufacturing a compound according to the second aspect of the present invention, detailed descriptions of parts that overlap with the first aspect of the present invention have been omitted, but even if such descriptions are omitted, the contents described in the first aspect of the present invention can be applied equally to the second aspect of the present invention.

[0115] According to one embodiment of the present invention, the above X 1 and X 2 One of them may be a halogen and the other may be B(OH)2, but is not limited thereto.

[0116] X of the above chemical formula 2 1 and X of the above chemical formula 3 2 A compound represented by Chemical Formula 4 can be prepared by reacting to form carbon-carbon bonds. At this time, X 1 and X 2 One of them may be a halogen and the other may be B(OH)2. For example, the above X 1 When using this halogen compound, the above X 2 The reaction can be carried out using a compound in which is B(OH)2, and conversely, the above X 2 In the case of a compound in which α is a halogen, the above X 1 The reaction can be carried out using this B(OH)2 compound.

[0117]

[0118] In addition, the third aspect of the present invention provides a composition for inhibiting HBV proliferation, comprising a compound according to the first aspect of the present invention as an active ingredient.

[0119] Regarding the composition for inhibiting HBV proliferation according to the third aspect of the present invention, detailed descriptions of parts that overlap with the first and / or second aspects of the present invention have been omitted, but even if such descriptions have been omitted, the contents described in the first and / or second aspects of the present invention may be applied equally to the third aspect of the present invention.

[0120] According to one embodiment of the present invention, HBc (Hepatitis B core) protein expression in HepG2 cells may be inhibited by a compound represented by the above chemical formula 1, but is not limited thereto.

[0121] The HBc protein is a core structural protein of the hepatitis B virus (HBV), essential for the formation of the viral capsid, and plays an important role in the replication and assembly processes of viral DNA.

[0122] HepG2 is a human liver cancer cell line and a widely used model system for HBV research. Because this cell line can express HBV proteins and produce viral particles, it is suitable for studying the effects of antiviral drugs.

[0123] The compound represented by Chemical Formula 1 according to the present invention can directly inhibit the expression of HBc protein or affect intracellular signaling pathways that regulate the expression of this protein, thereby inhibiting the formation of HBV capsid, which may make it difficult for the virus to replicate and assemble. This can consequently lead to a reduction in the production of new viral particles, which in turn leads to an effect of inhibiting the proliferation of HBV.

[0124] According to one embodiment of the present invention, the formation of nucleocapsid in HepG2 cells may be inhibited by the compound, but is not limited thereto.

[0125] The nucleocapsid is the core particle of HBV, having a structure in which the viral genome is packaged inside a capsid composed of HBc proteins. This is essential for the replication of viral DNA and the assembly of new viral particles.

[0126] Inhibition of nucleocapsid formation blocks important stages of the HBV life cycle, which can interfere with the packaging of the viral genome and the formation of mature viral particles.

[0127] Since this approach targets multiple stages of the viral life cycle simultaneously, effective antiviral activity can be expected, and it has the potential to reduce the likelihood of viral resistance developing.

[0128] According to one embodiment of the present invention, HBV DNA synthesis in HepG2 cells may be inhibited by the compound, but is not limited thereto.

[0129] According to one embodiment of the present invention, HBV DNA synthesis in Huh7 cells may be inhibited by the compound, but is not limited thereto.

[0130] HBV DNA synthesis is a critical step in the replication process of the hepatitis B virus (HBV), and inhibiting this step can lead to effects such as suppressing viral replication, preventing the spread of infection, reducing liver damage, and improving long-term prognosis.

[0131] Meanwhile, HepG2 cells are a human liver cancer cell line and a widely used model system for HBV research; the inhibition of HBV DNA synthesis in HepG2 cells directly demonstrates the antiviral effect of the compound. This cell line can mimic various stages of the HBV life cycle, making it useful for studying the mechanism of action of the compound.

[0132] Huh7 cells are also a human liver cancer cell line and another important model system used in HBV research; the inhibition of HBV DNA synthesis in Huh7 cells indicates that the compound's effect is not limited to a specific cell line. This suggests that the compound's antiviral effect may be more general and broad.

[0133] The compound represented by Chemical Formula 1 according to the present invention can directly inhibit the synthesis of HBV intermediate DNA or inhibit the activity of HBV DNA polymerase, and can also indirectly inhibit DNA synthesis by interfering with HBV capsid formation.

[0134] The consistent effect in two different cell lines suggests that the compound may be effective in various hepatocyte environments, allowing for the expectation of broad effects despite individual differences among patients.

[0135]

[0136] In addition, the fourth aspect of the present invention provides a pharmaceutical composition for preventing or treating HBV infection, comprising a composition for inhibiting HBV proliferation according to the third aspect of the present invention.

[0137] Regarding the pharmaceutical composition for preventing or treating HBV infection according to the fourth aspect of the present invention, detailed descriptions of parts that overlap with the first to third aspects of the present invention have been omitted, but even if such descriptions have been omitted, the contents described in the first to third aspects of the present invention may be applied equally to the fourth aspect of the present invention.

[0138] According to one embodiment of the present invention, the pharmaceutical composition for preventing or treating HBV infection may additionally comprise a pharmaceutically acceptable carrier, but is not limited thereto.

[0139] According to one embodiment of the present invention, the carrier may comprise, but is not limited to, a substance selected from the group consisting of lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, menthol, mineral oil, and combinations thereof.

[0140] According to one embodiment of the present invention, the pharmaceutical composition for preventing or treating HBV infection may additionally include a component selected from the group consisting of preservatives, solubilizers, stabilizers, humectants, sweeteners, coloring agents, flavoring agents, salts, buffers, antioxidants, lubricants, emulsifiers, suspending agents, preservatives, and combinations thereof, but is not limited thereto.

[0141] According to one embodiment of the present invention, the pharmaceutical composition for preventing or treating HBV infection may be administered by a method selected from the group consisting of intraperitoneal administration, oral administration, inhalation administration, intravenous administration, intramuscular administration, subcutaneous administration, dermal administration, intrauterine administration, tumor administration, rectal administration, and combinations thereof, but is not limited thereto.

[0142] The present invention is to be explained in more detail through the following examples, but the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

[0143]

[0144] [Example 1] ACT-C-023

[0145] (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)-N-(2-(dimethylamino)ethyl)acrylamide (JYPg37)

[0146] Oxalyl chloride (125 μL, 1.46 mmol) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (150 mg, 0.487 mmol) dissolved in dichloromethane. After heating at reflux temperature for 3 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of N,N-dimethylethylenediamine (63.8 μL, 0.584 mmol) and DIPEA (178 μL, 1.02 mmol) at 0°C. The reaction mixture was stirred at room temperature for 18 hours. After the reaction was complete, ethyl acetate and water were poured into the solution. The organic layer was washed with water, dried with Na2SO4, filtered, and concentrated. The residue was purified by silica gel column chromatography to prepare a compound represented by the following structural formula. (42 mg, 22.8%)

[0147]

[0148] Figure 1 is the NMR spectrum of a compound according to Example 1 of the present invention.

[0149] 1 H NMR (600 MHz, CDCl3) δ 8.16 (d,J = 2.1 Hz, 1H), 8.06 (s, 1H), 7.44 (d,J = 4.1 Hz, 1H), 7.41 (d,J = 9.0 Hz, 1H), 7.28 (d,J = 2.8 Hz, 1H), 7.21 (d,J = 3.4 Hz, 1H), 7.12-7.05 (m, 1H), 3.50 (q,J = 5.7 Hz, 2H), 2.53 (t,J = 5.9 Hz, 2H), 2.30 (s, 6H).

[0150] [Example 2] ACT-C-006

[0151] (E)-N-(3-chlorophenethyl)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylamide (JYPf76)

[0152] Oxalyl chloride (62.7 μL, 0.730 mmol) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (75 mg, 0.243 mmol) dissolved in dichloromethane. After heating to reflux temperature for 3 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of 2-(3-chlorophenyl)ethylamine (40.6 μL, 0.292 mmol) and DIPEA (89.0 μL, 0.511 mmol) at 0°C. The reaction mixture was stirred at room temperature for 18 hours. After the reaction was complete, ethyl acetate and water were poured into the solution. The organic layer was washed with a saturated NaHCO3 solution, a 1 N HCl solution, and brine, dried with Na2SO4, filtered, and concentrated. The residue was purified by silica gel column chromatography to prepare a yellow powder of the compound represented by the following structural formula. (17.6 mg, 16.3%)

[0153]

[0154] Figure 2 is the NMR spectrum of a compound according to Example 2 of the present invention. 1 H NMR (600 MHz, CDCl3) δ 8.12 (d,J = 2.1 Hz, 1H), 8.06 (s, 1H), 7.44 (d,J = 4.1 Hz, 1H), 7.41 (d,J = 9.0 Hz, 1H), 7.28 (dd,J = 8.3, 2.8 Hz, 2H), 7.23 (dd,J = 12.7, 3.8 Hz, 3H), 7.12 (d,J = 7.6 Hz, 1H), 6.41 (s, 1H), 3.67 (q,J = 6.7 Hz, 2H), 2.90 (t,J = 6.9 Hz, 2H).

[0155] [Example 3] ACT-C-010

[0156] (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)-N-ethylacrylamide (JYPg39)

[0157] Oxalyl chloride (125 μL, 1.46 mmol) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (150 mg, 0.487 mmol) dissolved in dichloromethane. After heating to reflux temperature for 3 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of ethylamine hydrochloride (47.6 mg, 0.584 mmol) and DIPEA (263 μL, 1.51 mmol) at 0°C. The reaction mixture was stirred at room temperature for 18 hours. After the reaction was complete, ethyl acetate and water were poured into the solution. The organic layer was washed with water, dried with Na2SO4, filtered, and concentrated. The residue was purified by silica gel column chromatography. (5 mg, 3.1%)

[0158]

[0159] Figure 3 is the NMR spectrum of a compound according to Example 3 of the present invention.

[0160] 1 H NMR (600 MHz, CDCl3+ CD3OD) δ 8.13 (d,J = 2.8 Hz, 1H), 8.04 (s, 1H), 7.47 (d,J = 3.4 Hz, 1H), 7.43 (d,J = 8.3 Hz, 1H), 7.39 (s, 1H), 7.30 (dd,J = 8.6, 2.4 Hz, 1H), 7.26 (d,J = 3.4 Hz, 1H), 3.46 (q,J = 7.3 Hz, 2H), 1.26 (t,J = 7.2 Hz, 3H).

[0161] [Example 4] ACT-C-012

[0162] (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)-N-hexylacrylamide (JYPf90)

[0163] Oxalyl chloride (62.7 μL, 0.730 mmol) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (75 mg, 0.243 mmol) dissolved in dichloromethane. After heating to reflux temperature for 3 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of hexylamine (39.4 μL, 0.292 mmol) and DIPEA (89.0 μL, 0.511 mmol) at 0°C. The reaction mixture was stirred at room temperature for 18 hours. After the reaction was complete, ethyl acetate and water were poured into the solution. The organic layer was washed with water, dried with Na2SO4, filtered, and concentrated. The residue was purified by silica gel column chromatography to obtain a pale yellow powder. (17.6 mg, 18.5%)

[0164]

[0165] Figure 4 is the NMR spectrum of a compound according to Example 4 of the present invention.

[0166] 1 H NMR (600 MHz, CDCl3) δ 8.14 (d,J = 2.1 Hz, 1H), 8.07 (s, 1H), 7.40-7.44 (m, 2H), 7.28 (dd,J = 8.6, 2.4 Hz, 1H), 7.21 (d,J = 4.1 Hz, 1H), 6.35 (s, 1H), 3.43 (q,J = 6.7 Hz, 2H), 1.60 (q,J = 7.3 Hz, 2H), 1.31-1.39 (m, 6H), 0.90 (t,J = 6.9 Hz, 3H)

[0167] [Example 5] ACT-C-020

[0168] (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)-N-(2-hydroxyethyl)acrylamide (JYPg17)

[0169] Oxalyl chloride (125 μL, 1.46 mmol) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (150 mg, 0.487 mmol) dissolved in dichloromethane. After heating to reflux temperature for 3 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of 2-ethanolamine (35.0 μL, 0.584 mmol) and DIPEA (178 μL, 1.02 mmol) at 0°C. The reaction mixture was stirred at room temperature for 18 hours. After the reaction was complete, ethyl acetate and water were poured into the solution. The organic layer was washed with water, dried with Na2SO4, filtered, and concentrated. The residue was purified by silica gel column chromatography to obtain a pale yellow powder. (23 mg, 13.5%)

[0170]

[0171] Figure 5 is the NMR spectrum of a compound according to Example 5 of the present invention.

[0172] 1 H NMR (600 MHz, DMSO-d6) δ 8.31 (t,J = 5.5 Hz, 1H), 8.11 (d,J = 2.8 Hz, 1H), 8.07 (s, 1H), 7.67 (d,J = 8.3 Hz, 1H), 7.57 (d,J = 3.4 Hz, 1H), 7.53 (dd,J = 8.6, 2.4 Hz, 1H), 7.49 (d,J = 3.4 Hz, 1H), 4.78 (t,J = 5.5 Hz, 1H), 3.49 (q,J = 6.0 Hz, 2H), 3.29 (q,J = 5.3 Hz, 2H), 3.17 (s, 1H), 2.97 (s, 1H)

[0173] [Example 6] ACT-C-021

[0174] (E)-tert-butyl (2-(2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylamido)ethyl)carbamate (JYPg23)

[0175] Oxalyl chloride (125 μL, 1.46 mmol) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (150 mg, 0.487 mmol) dissolved in dichloromethane. After heating to reflux temperature for 3 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of N-(tert-butoxycarbonyl)-1,2-diaminoethane (92.7 μL, 0.584 mmol) and DIPEA (178 μL, 1.02 mmol) at 0°C. The reaction mixture was stirred at room temperature for 18 hours. After the reaction was complete, the formed solid was filtered to obtain the target compound (67 mg, 30.6%).

[0176]

[0177] Figure 6 is the NMR spectrum of a compound according to Example 6 of the present invention.

[0178] 1 H NMR (600 MHz, DMSO-d6) δ 8.12 (d,J = 2.8 Hz, 1H), 7.76 (s, 1H), 7.64 (d,J = 9.0 Hz, 1H), 7.47-7.49 (m, 2H), 7.25 (d,J = 3.4 Hz, 1H), 7.02 (s, 1H), 3.17 (q,J = 6.0 Hz, 2H), 2.82 (t,J = 6.5 Hz, 2H), 1.39 (s, 9H).

[0179] [Example 7] ACT-C-026

[0180] (E)-2-cyano-N-(cyclohexylmethyl)-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylamide (JYPf96)

[0181] Oxalyl chloride (125 μL, 1.46 mmol) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (150 mg, 0.487 mmol) dissolved in dichloromethane. After heating to reflux temperature for 3 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of cyclohexanedmethylamine (80.6 μL, 0.584 mmol) and DIPEA (178 μL, 1.02 mmol) at 0°C. The reaction mixture was stirred at room temperature for 18 hours. After the reaction was complete, ethyl acetate and water were poured into the solution. The organic layer was washed with water, dried with Na2SO4, filtered, and concentrated. The residue was purified by silica gel column chromatography to obtain a pale yellow powder. (68.7 mg, 35.0%)

[0182]

[0183] Figure 7 is the NMR spectrum of a compound according to Example 7 of the present invention.

[0184] 1 H NMR (600 MHz, CDCl3) δ 8.14 (d,J = 2.8 Hz, 1H), 8.06 (s, 1H), 7.44 (d,J = 4.1 Hz, 1H), 7.41 (d,J = 8.3 Hz, 1H), 7.28 (dd, J = 8.6, 2.4 Hz, 1H), 7.20 (d,J = 4.1 Hz, 1H), 6.42 (s, 1H), 3.28 (t,J = 6.5 Hz, 2H), 1.74-1.78 (m, 4H), 1.67-1.70 (m, 1H), 1.16-1.29 (m, 4H), 0.96-1.02 (m, 2H)

[0185] [Example 8] ACT-C-028

[0186] (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)-N-(4-fluorobenzyl)acrylamide (Kma1047)

[0187] Oxalyl chloride (63.0 μL, 0.738 mmol) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (76.0 mg, 0.246 mmol) dissolved in dichloromethane. After heating to reflux temperature for 3 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of 4-fluorobenzylamine (34 μL, 0.295 mmol) and DIPEA (86.0 μL, 0.492 mmol) at 0°C. The reaction mixture was stirred at room temperature for 18 hours. After the reaction was complete, ethyl acetate and water were poured into the solution. The organic layer was washed with water, dried with Na2SO4, filtered, and concentrated. The residue was purified by silica gel column chromatography to obtain a pale yellow powder. (60 mg, 60.0%)

[0188]

[0189] Figure 8 is the NMR spectrum of a compound according to Example 8 of the present invention.

[0190] 1 H-NMR (600 MHz, CDCl3) δ 8.09 (d,J = 2.8 Hz, 1H), 8.08 (s, 1H), 7.43 (d,J = 4.1 Hz, 1H), 7.38 (d,J = 9.0 Hz, 1H), 7.31 (q,J = 4.6 Hz, 2H), 7.25 (dd,J = 8.6, 2.4 Hz, 1H), 7.20 (d,J = 3.4 Hz, 1H), 7.04 (t,J = 8.6 Hz, 2H), 6.75 (t,J = 5.2 Hz, 1H), 4.57 (d,J = 6.2 Hz, 2H)

[0191] [Example 9] ACT-C-029

[0192] (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)-N-(4-(trifluoromethyl)benzyl)acrylamide (KMa1051)

[0193] Oxalyl chloride (125 μL, 1.46 mmol) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (150 mg, 0.487 mmol) dissolved in dichloromethane. After heating to reflux temperature for 3 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of 4-(trifluoromethyl)benzylamine (83 μL, 0.584 mmol) and DIPEA (170 μL, 0.974 mmol) at 0°C. The reaction mixture was stirred at room temperature for 18 hours. After the reaction was complete, ethyl acetate and water were poured into the solution. The organic layer was washed with water, dried with Na2SO4, filtered, and concentrated. The residue was purified by silica gel column chromatography to obtain a pale yellow powder. (70.6 mg, 39.0%)

[0194]

[0195] Figure 9 is the NMR spectrum of a compound according to Example 9 of the present invention.

[0196] 1 H-NMR (600 MHz, CDCl3) δ 8.08 (s, 2H), 7.60 (d,J = 7.6 Hz, 2H), 7.43-7.45 (m, 3H), 7.39 (d,J = 9.0 Hz, 1H), 7.26 (dd,J = 8.3, 2.1 Hz, 1H), 7.20 (d,J = 3.4 Hz, 1H), 6.94 (t,J = 5.9 Hz, 1H), 4.66 (d,J = 5.5 Hz, 2H)

[0197] [Example 10] ACT-C-030

[0198] (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)-N-(3-(trifluoromethyl)benzyl)acrylamide (KMa1052)

[0199] Oxalyl chloride (125 μL, 1.46 mmol) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (150 mg, 0.487 mmol) dissolved in dichloromethane. After heating to reflux temperature for 3 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of 3-(trifluoromethyl)benzylamine (84.0 μL, 0.584 mmol) and DIPEA (170 μL, 0.974 mmol) at 0°C. The reaction mixture was stirred at room temperature for 18 hours. After the reaction was complete, ethyl acetate and water were poured into the solution. The organic layer was washed with water, dried with Na2SO4, filtered, and concentrated. The residue was purified by silica gel column chromatography to obtain a pale yellow powder. (51.2 mg, 22.8%)

[0200]

[0201] Figure 10 is the NMR spectrum of a compound according to Example 10 of the present invention.

[0202] 1 H-NMR (600 MHz, CDCl3) δ 8.12 (d,J = 2.8 Hz, 1H), 8.10 (d,J = 5.5 Hz, 1H), 7.58 (d,J = 10.3 Hz, 2H), 7.53 (d,J = 7.6 Hz, 1H), 7.49 (t, J = 7.6 Hz, 1H), 7.43-7.46 (m, 1H), 7.40-7.42 (m, 1H), 7.28 (dd,J = 8.6, 2.4 Hz, 1H), 7.24 (d, J = 4.1 Hz, 1H), 6.79 (t, J = 5.9 Hz, 1H), 4.67 (d, J = 6.2 Hz, 2H)

[0203] [Example 11] ACT-C-031

[0204] (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)-N-(4-(trifluoromethoxy)benzyl)acrylamide (KMa1053)

[0205] Oxalyl chloride (67.0 μL, 0.780 mmol) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (80.0 mg, 0.260 mmol) dissolved in dichloromethane. After heating to reflux temperature for 3 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of 4-(trifluoromethoxy)benzylamine (47.0 μL, 0.311 mmol) and DIPEA (90.0 μL, 0.520 mmol) at 0°C. The reaction mixture was stirred at room temperature for 18 hours. After the reaction was complete, ethyl acetate and water were poured into the solution. The organic layer was washed with water, dried with Na2SO4, filtered, and concentrated. The residue was purified by silica gel column chromatography to obtain a light yellow powder. (63.7 mg, 50.9%)

[0206]

[0207] Figure 11 is the NMR spectrum of a compound according to Example 11 of the present invention.

[0208] 1 H-NMR (600 MHz, CDCl3) δ 8.09-8.10 (m, 2H), 7.44 (d,J = 4.1 Hz, 1H), 7.39 (d, J = 9.0 Hz, 1H), 7.37 (d,J = 8.3 Hz, 2H), 7.25-7.27 (m, 1H), 7.20-7.22 (m, 3H), 6.79 (t, J = 5.9 Hz, 1H), 4.61 (d, J = 5.5 Hz, 2H)

[0209] [Example 12] ACT-C-032

[0210] (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)-N-(3,4-dimethoxybenzyl)acrylamide (KMb1001)

[0211] Oxalyl chloride (334 μL, 3.90 mmol) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (400 mg, 1.30 mmol) dissolved in dichloromethane. After heating at reflux temperature for 3 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of 3,4-dimethoxybenzylamine (235 μL, 1.60 mmol) and DIPEA (452 ​​μL, 2.60 mmol) at 0°C. The reaction mixture was stirred at room temperature for 18 hours. After the reaction was complete, the residue was purified by silica gel column chromatography to obtain a pale yellow powder (121 mg, 20.4%).

[0212]

[0213] Figure 12 is the NMR spectrum of a compound according to Example 12 of the present invention.

[0214] 1 H-NMR (600 MHz, CDCl3) δ 8.08 (d, J = 3.4 Hz, 2H), 7.42 (d,J = 4.1 Hz, 1H), 7.37 (d, J = 8.3 Hz, 1H), 7.24 (dd,J = 8.6, 2.4 Hz, 1H), 7.19 (d, J = 4.1 Hz, 1H), 6.89 (dd, J = 8.3, 2.1 Hz, 1H), 6.86 (d, J = 1.0 Hz, 1H), 6.83 (d,J = 7.6 Hz, 1H), 6.73 (t, J = 5.5 Hz, 1H), 4.54 (d,J = 5.5 Hz, 2H), 3.89 (d, J = 9.0 Hz, 3H), 3.87 (d, J = 8.3 Hz, 3H)

[0215] [Example 13] ACT-C-033

[0216] (E)-2-cyano-N-(3,4-dichlorobenzyl)-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylamide (KMb1003)

[0217] Oxalyl chloride (108 μL, 1.27 mmol) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (130 mg, 0.420 mmol) dissolved in dichloromethane. After heating at reflux temperature for 3 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of 3,4-dichlorobenzylamine (67.0 μL, 0.500 mmol) and DIPEA (146 μL, 0.840 mmol) at 0°C. The reaction mixture was stirred at room temperature for 18 hours. After the reaction was complete, the residue was purified by silica gel column chromatography to obtain a pale yellow powder (51.2 mg, 26.1%).

[0218]

[0219] Figure 13 is the NMR spectrum of a compound according to Example 13 of the present invention.

[0220] 1 H-NMR (600 MHz, CDCl3) δ 8.10 (d, J = 2.1 Hz, 1H), 8.08 (d, J = 4.8 Hz, 1H), 7.44 (t,J = 4.5 Hz, 1H), 7.42-7.43 (m, 1H), 7.40-7.42 (m, 2H), 7.27 (dd, J = 9.0, 2.8 Hz, 1H), 7.23 (d,J = 4.1 Hz, 1H), 7.17 (dd,J = 8.3, 2.1 Hz,1H), 6.83 (t,J = 5.9 Hz, 1H), 4.56 (d,J = 6.2 Hz, 2H)

[0221] [Example 14] ACT-C-045

[0222] Ethyl (E)-(2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acryloyl)glycinate (JYPh45)

[0223] Oxalyl chloride (125 μL, 1.46 mmol) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (150 mg, 0.487 mmol) dissolved in dichloromethane. After heating to reflux temperature for 3 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of glycine ethyl ester hydrochloride (81.5 mg, 0.584 mmol) and DIPEA (263 μL, 1.51 mmol) at 0°C. The reaction mixture was stirred at room temperature for 18 hours. After the reaction was complete, ethyl acetate and water were poured into the solution. The organic layer was dried with Na2SO4, filtered, and concentrated. The residue was purified by silica gel column chromatography to obtain the target compound. (75.0 mg, 39.2%)

[0224]

[0225] Figure 14 is the NMR spectrum of a compound according to Example 14 of the present invention.

[0226] 1 H NMR (600 MHz, CDCl3) δ 8.14 (d,J = 2.1 Hz, 1H), 8.07 (s, 1H), 7.46 (d,J = 4.1 Hz, 1H), 7.41 (d,J = 9.0 Hz, 1H), 7.28 (dd,J = 8.6, 2.4 Hz, 1H), 7.25 (d,J = 3.4 Hz, 1H), 6.88 (d,J = 4.8 Hz, 1H), 4.27 (q,J = 7.1 Hz, 2H), 4.20 (d,J = 4.8 Hz, 2H), 1.32 (t,J = 7.2 Hz, 3H).

[0227] [Example 15] ACT-C-049

[0228] (E)-2-cyano-N-(2-cyclohexylethyl)-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylamide (YJb29)

[0229] Oxalyl chloride (125 μL, 1.46 mmol, 3 equivalents) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (150 mg, 0.487 mmol, 1 equivalent) dissolved in dichloromethane. After heating to reflux temperature for 4 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of 2-cyclohexylethylamine (85 μL, 0.584 mmol, 1.2 equivalents) and DIPEA (178 μL, 1.02 mmol, 2 equivalents) at 0°C. The reaction mixture was stirred at room temperature for 23 hours. After the reaction was complete, EA and water were poured into the solution. The organic layer was washed with water, dried with Na2SO4, filtered, and concentrated. The product was purified by flash column chromatography (EA / Hex; 5:95 to 9:91) to obtain the target compound (67 mg, 33%).

[0230]

[0231] Figure 15 is the NMR spectrum of a compound according to Example 15 of the present invention.

[0232] 1 H NMR (600 MHz, CDCl3) δ 8.14 (d,J = 2.5 Hz, 1H), 8.06 (s, 1H), 7.44 (d,J = 3.8 Hz, 1H), 7.41 (d,J = 8.6 Hz, 1H), 7.28 (dd,J = 8.6, 2.5 Hz, 1H), 7.20 (d,J = 3.7 Hz, 1H), 6.31 (s, 1H), 3.47-3.43 (m, 2H), 1.78-1.70 (m, 4H), 1.67 (d,J = 12.1 Hz,1H), 1.53-1.48 (m, 2H), 1.35 (ddt,J = 14.5, 7.5, 3.5 Hz, 1H), 1.29-1.14 (m, 3H), 1.00-0.91 (m, 2H).

[0233] [Example 16] ACT-C-051

[0234] (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)-N-isobutylacrylamide (YJb57)

[0235] Oxalyl chloride (125 μL, 1.46 mmol, 3 equivalents) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (150 mg, 0.487 mmol, 1 equivalent) dissolved in dichloromethane. After heating to reflux temperature for 4 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of isobutylamine (59 μL, 0.584 mmol, 1.2 equivalents) and DIPEA (178 μL, 1.02 mmol, 2 equivalents) at 0°C. The reaction mixture was stirred at room temperature for 23 hours. After the reaction was complete, EA and water were poured into the solution. The organic layer was washed with water, dried with Na2SO4, filtered, and concentrated. The product was purified by flash column chromatography (EA / Hex; 10:90 to 12:88) to obtain the target compound (111 mg, 63%).

[0236]

[0237] Figure 16 is the NMR spectrum of a compound according to Example 16 of the present invention.

[0238] 1 H NMR (600 MHz, CDCl3) δ 8.14 (d,J = 2.6 Hz, 1H), 8.06 (s, 1H), 7.44 (d,J = 3.8 Hz, 1H), 7.40 (d,J = 8.6 Hz, 1H), 7.29-7.27 (m, 1H), 7.20 (d,J = 3.9 Hz, 1H), 6.45-6.41 (m, 1H), 3.27 (dd,J = 6.9, 6.0 Hz, 2H), 1.90 (hept,J = 6.8 Hz, 1H), 0.98 (d,J = 6.7 Hz, 6H).

[0239] [Example 17] ACT-C-053

[0240] (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)-N-pentylacrylamide (YJb59)

[0241] Oxalyl chloride (125 μL, 1.46 mmol, 3 equivalents) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (150 mg, 0.487 mmol, 1 equivalent) dissolved in dichloromethane. After heating to reflux temperature for 4 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of amylamine (68 μL, 0.584 mmol, 1.2 equivalents) and DIPEA (178 μL, 1.02 mmol, 2 equivalents) at 0°C. The reaction mixture was stirred at room temperature for 23 hours. After the reaction was complete, EA and water were poured into the solution. The organic layer was washed with water, dried with Na2SO4, filtered, and concentrated. The product was purified by flash column chromatography (EA / Hex; 4:96 to 20:80) to obtain the target compound (60 mg, 33%).

[0242]

[0243] Figure 17 is the NMR spectrum of a compound according to Example 17 of the present invention.

[0244] 1 H NMR (600 MHz, CDCl3) δ 8.13 (d,J = 2.5 Hz, 1H), 8.05 (s, 1H), 7.43 (d,J = 3.8 Hz, 1H), 7.40 (d,J = 8.6 Hz, 1H), 7.26 (dd,J = 8.6, 2.5 Hz, 1H), 7.19 (d,J = 3.8 Hz, 1H), 6.35 (s, 1H), 3.43-3.39 (m, 2H), 1.60 (p,J = 7.3 Hz, 2H), 1.37-1.33 (m, 4H), 0.94-0.88 (m, 3H).

[0245] [Example 18] ACT-C-055

[0246] (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)-N-(2-methoxybenzyl)acrylamide (YJb39)

[0247] Oxalyl chloride (125 μL, 1.46 mmol, 3 equivalents) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (150 mg, 0.487 mmol, 1 equivalent) dissolved in dichloromethane. After heating to reflux temperature for 4 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of 2-methoxybenzylamine (76 μL, 0.584 mmol, 1.2 equivalents) and DIPEA (178 μL, 1.02 mmol, 2 equivalents) at 0°C. The reaction mixture was stirred at room temperature for 23 hours. After the reaction was complete, EA and water were poured into the solution. The organic layer was washed with water, dried with Na2SO4, filtered, and concentrated. The product was purified by flash column chromatography (EA / Hex; 4:96 to 15:85) to obtain the target compound (13 mg, 6%).

[0248]

[0249] Figure 18 is the NMR spectrum of a compound according to Example 18 of the present invention.

[0250] 1 H NMR (600 MHz, CDCl3) δ 8.12 (d,J = 2.6 Hz, 1H), 8.05 (s, 1H), 7.42 (d,J = 3.8 Hz, 1H), 7.39 (d,J = 8.5 Hz, 1H), 7.30 (dd,J = 7.3, 2.3 Hz, 2H), 7.27 (dd,J = 11.2, 2.2 Hz, 1H), 7.17 (d,J = 3.8 Hz, 1H), 7.10 (t,J = 5.6 Hz, 1H), 6.95-6.90 (m, 2H), 4.59 (d,J = 5.9 Hz, 2H), 3.91 (s, 3H).

[0251] [Example 19] ACT-C-057

[0252] (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)-N-(3,4-dimethoxybenzyl)acrylamide (YJb07)

[0253] Oxalyl chloride (125 μL, 1.46 mmol, 3 equivalents) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (150 mg, 0.487 mmol, 1 equivalent) dissolved in dichloromethane. After heating to reflux temperature for 4 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of 3,4-dimethoxybenzylamine (88 μL, 0.584 mmol, 1.2 equivalents) and DIPEA (178 μL, 1.02 mmol, 2 equivalents) at 0°C. The reaction mixture was stirred at room temperature for 24 hours. After the reaction was complete, EA and water were poured into the solution. The organic layer was washed with water, dried with Na2SO4, filtered, and concentrated. The product was purified by flash column chromatography (EA / Hex; 5:95 to 19:81) to obtain the target compound (33 mg, 15%).

[0254]

[0255] Figure 19 is the NMR spectrum of a compound according to Example 19 of the present invention.

[0256] 1 H NMR (600 MHz, CDCl3) δ 8.13 (d,J = 2.5 Hz, 1H), 8.11 (s, 1H), 7.45 (d,J = 3.8 Hz, 1H), 7.41 (d,J = 8.6 Hz, 1H), 7.28 (d,J = 2.5 Hz, 1H), 7.23 (d,J = 3.8 Hz, 1H), 6.89 (dd,J = 8.3, 1.9 Hz, 1H), 6.85 (d,J = 8.2 Hz, 2H), 6.61 (s, 1H), 4.55 (d,J = 5.7 Hz, 2H), 3.89 (d,J = 5.9 Hz, 6H).

[0257] [Example 20] ACT-C-059

[0258] (E)-N-(3-chlorobenzyl)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylamide (YJb26)

[0259] Oxalyl chloride (125 μL, 1.46 mmol, 3 equivalents) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (150 mg, 0.487 mmol, 1 equivalent) dissolved in dichloromethane. After heating to reflux temperature for 4 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of 3-chlorobenzylamine (53 μL, 0.584 mmol, 1.2 equivalents) and DIPEA (178 μL, 1.02 mmol, 2 equivalents) at 0°C. The reaction mixture was stirred at room temperature for 23 hours. After the reaction was complete, EA and water were poured into the solution. The organic layer was washed with water, dried with Na2SO4, filtered, and concentrated. The product was purified by flash column chromatography (EA / Hex; 5:95 to 9:91) to obtain the target compound (58 mg, 28%).

[0260]

[0261] FIG. 20 is the NMR spectrum of a compound according to Example 20 of the present invention.

[0262] 1 H NMR (600 MHz, CDCl3) δ 8.13 (d,J = 2.5 Hz, 1H), 8.11 (s, 1H), 7.45 (d,J = 3.9 Hz, 1H), 7.41 (d,J = 8.6 Hz, 1H), 7.32 (s, 1H), 7.30-7.28 (m, 3H), 7.27 (d,J = 2.5 Hz, 1H), 7.24 (d,J = 3.8 Hz, 1H), 7.21 (dd,J = 7.2, 1.5 Hz, 1H), 6.72-6.69 (m, 1H), 4.59 (d,J = 5.9 Hz, 2H).

[0263] [Example 21] ACT-C-060

[0264] (E)-N-(4-chlorobenzyl)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylamide (YJb37)

[0265] Oxalyl chloride (125 μL, 1.46 mmol, 3 equivalents) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (150 mg, 0.487 mmol, 1 equivalent) dissolved in dichloromethane. After heating to reflux temperature for 4 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of 4-chlorobenzylamine (71 μL, 0.584 mmol, 1.2 equivalents) and DIPEA (178 μL, 1.02 mmol, 2 equivalents) at 0°C. The reaction mixture was stirred at room temperature for 23 hours. After the reaction was complete, EA and water were poured into the solution. The organic layer was washed with water, dried with Na2SO4, filtered, and concentrated. The product was purified by flash column chromatography (EA / Hex; 3:97 to 11:88) to obtain the target compound (76 mg, 36%).

[0266]

[0267] Figure 21 is the NMR spectrum of a compound according to Example 21 of the present invention.

[0268] 1 H NMR (600 MHz, CDCl3) δ 8.13 (d,J = 2.5 Hz, 1H), 8.10 (s, 1H), 7.45 (d,J = 3.8 Hz, 1H), 7.41 (d,J = 8.5 Hz, 1H), 7.35-7.32 (m, 2H), 7.29-7.26 (m, 4H), 7.23 (d,J = 3.7 Hz, 1H), 6.68 (t,J = 5.5 Hz, 1H), 4.58 (d,J = 5.9 Hz,2H).

[0269] [Example 22] ACT-C-061

[0270] (E)-N-(4-chlorophenethyl)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylamide (YJb14)

[0271] Oxalyl chloride (125 μL, 1.46 mmol, 3 equivalents) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (150 mg, 0.487 mmol, 1 equivalent) dissolved in dichloromethane. After heating to reflux temperature for 4 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of 2-(4-chlorophenyl)ethylamine (81 μL, 0.584 mmol, 1.2 equivalents) and DIPEA (178 μL, 1.02 mmol, 2 equivalents) at 0°C. The reaction mixture was stirred at room temperature for 23 hours. After the reaction was complete, EA and water were poured into the solution. The organic layer was washed with water, dried with Na2SO4, filtered, and concentrated. The product was purified by flash column chromatography (EA / Hex; 5:95 to 17:83) to obtain the target compound (63 mg, 29%).

[0272]

[0273] Figure 22 is the NMR spectrum of a compound according to Example 22 of the present invention.

[0274] 1 H NMR (600 MHz, CDCl3) δ 8.11 (d,J = 2.5 Hz, 1H), 8.04 (s, 1H), 7.44 (d,J = 3.8 Hz, 1H), 7.40 (d,J = 8.6 Hz, 1H), 7.31-7.29 (m, 2H), 7.27 (dd,J = 8.6, 2.5 Hz, 1H), 7.20 (d,J = 3.8 Hz, 1H), 7.16 (d,J = 8.4 Hz, 2H), 6.38 (s, 1H), 3.68-3.62 (m, 2H), 2.88 (t,J = 7.3 Hz, 2H).

[0275] [Example 23] ACT-C-064

[0276] (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)-N-(3-methoxybenzyl)acrylamide (YJb40)

[0277] Oxalyl chloride (125 μL, 1.46 mmol, 3 equivalents) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (150 mg, 0.487 mmol, 1 equivalent) dissolved in dichloromethane. After heating to reflux temperature for 4 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of 3-methoxybenzylamine (75 μL, 0.584 mmol, 1.2 equivalents) and DIPEA (178 μL, 1.02 mmol, 2 equivalents) at 0°C. The reaction mixture was stirred at room temperature for 23 hours. After the reaction was complete, EA and water were poured into the solution. The organic layer was washed with water, dried with Na2SO4, filtered, and concentrated. The product was purified by flash column chromatography (EA / Hex; 4:96 to 20:80) to obtain the target compound (73 mg, 35%).

[0278]

[0279] FIG. 23 is the NMR spectrum of a compound according to Example 23 of the present invention.

[0280] 1 H NMR (600 MHz, CDCl3) δ 8.12 (d,J = 2.5 Hz, 1H), 8.10 (s, 1H), 7.44 (d,J = 3.8 Hz, 1H), 7.40 (d,J = 8.5 Hz, 1H), 7.29-7.25 (m, 2H), 7.22 (d,J = 3.7 Hz, 1H), 6.90 (d,J = 7.9 Hz, 1H), 6.86-6.83 (m, 2H), 6.64 (s, 1H), 4.58 (d,J = 5.7 Hz, 2H), 3.81 (s, 3H).

[0281] [Example 24] ACT-C-065

[0282] (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)-N-propylpropanamide (YJb66)

[0283] Oxalyl chloride (125 μL, 1.46 mmol, 3 equivalents) was slowly added to a solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (150 mg, 0.487 mmol, 1 equivalent) dissolved in dichloromethane. After heating to reflux temperature for 4 hours, the solution was evaporated under reduced pressure. The residue was dissolved in dichloromethane and then added to a dichloromethane solution of propylamine (48 μL, 0.584 mmol, 1.2 equivalents) and DIPEA (178 μL, 1.02 mmol, 2 equivalents) at 0°C. The reaction mixture was stirred at room temperature for 18 hours. After the reaction was complete, EA and water were poured into the solution. The organic layer was washed with water, dried with Na2SO4, filtered, and concentrated. The product was purified by flash column chromatography (EA / Hex; 0:100 to 30:70) to obtain the target compound. (96.7 mg, 56.9%)

[0284]

[0285] Figure 24 is the NMR spectrum of a compound according to Example 24 of the present invention.

[0286] 1 H NMR (600 MHz, CDCl3) δ 8.14 (d,J = 2.5 Hz, 1H), 8.07 (s, 1H), 7.44 (d,J = 3.8 Hz, 1H), 7.41 (d,J = 8.6 Hz, 1H), 7.28 (dd,J = 8.6, 2.5 Hz, 1H), 7.21 (d,J = 3.8 Hz, 1H), 6.38 (s, 1H), 3.40 (q,J = 3.4 Hz, 2H), 1.68 - 1.61 (m, 2H), 0.99 (t,J = 7.4 Hz, 3H).

[0287]

[0288] [Experimental Example 1] MTS assay: Cytotoxicity study

[0289] 2 x 10 per well in a 96-well microtiter plate 4 / well HepG2 cells were seeded with 100 μL of cell culture medium and cultured under 5% CO2 at 37°C. The following day, compounds were treated at various concentrations (1, 5, 10, 50, and 100 μM), and the cells were cultured for a total of 48 hours. Subsequently, 10 μL of MTS reagent was added to each well, and the cells were cultured for 4 hours under standard cell culture conditions at 37°C. After briefly shaking the plates on a shaker, absorbance was measured at a plate reader OD of 490 nm. For Examples 14–24, the cells were cultured for a total of 72 hours.

[0290] Figures 25a and 25b show the results of a cytotoxicity test according to one experimental example of the present invention. Referring to Figures 25a and 25b, the cytotoxicity of the compound according to Example 1 was measured at 1, 5, 10, 50, and 100 μM in HepG2 liver cancer cell lines, and as a result, it showed no cytotoxicity at 1, 5, and 10 μM, and CC 50 With values ​​ranging from 25.3 to 45.1 μM, there was no cytotoxicity at the concentrations used in the experiment. In addition, other compounds also showed little to no cytotoxicity, or CC 50 The value was a very high concentration.

[0291] [Experimental Example 2]

[0292] An experiment was performed to inhibit HBV proliferation in liver cancer cell lines using the compound according to Example 1.

[0293] After seeding HepG2 and Huh7 cells, 8 μg of 1.3mer HBV WT plasmid DNA was transfected into HepG2 cells and 3 μg into Huh7 cells the following day. pGEM4Z was used as a control. Simultaneously with transfection, HepG2 cells (2 x 10⁶ 6 / 6cm) Compound 1 at a concentration of 10μM was applied to Huh7 cells (5x10 5In the / 6cm cell group, Compound 1 was treated at a concentration of 5 μM, and on the following day (24 hours later), when the medium was changed, Compound 1 was treated at the same concentration as before and cultured for an additional 48 hours. As a control, AGK2 was treated at a concentration of 10 μM in HepG2 cells and 2.5 μM in Huh7 cells and cultured in the same manner. After culturing for a total of 72 hours following transfection, cell lysates were prepared with 0.2% NP40-NTE buffer, followed by SDS-10% PAGE and Western blotting. Anti-acetylated α-tubulin, anti-α-tubulin, anti-sirtuin 2, anti-HBc, and anti-GAPDH antibodies were used as primary antibodies for Western blotting, and enhanced chemiluminescence (ECL) was observed. After incubating for a total of 72 hours following transfection, cell lysate was prepared with 0.2% NP40-NTE buffer, subjected to SDS-10% PAGE, and Western blotting with each antibody. Here, Western blotting was performed using anti-acetylated α-tubulin, anti-α-tubulin, anti-sirtuin 2, anti-HBc, and anti-GAPDH antibodies, and observed via enhanced chemiluminescence (ECL). After 1% native agarose gel electrophoresis (NAGE) of the lysate, Western blotting (immunoblotting) was performed using an anti-HBc antibody, and core particle formation was observed via ECL. To observe the synthesis of HBV replicative intermediate DNA (RI DNA) via Southern blotting, HBV DNA was extracted from the isolated core particles, subjected to 1% NAGE, and then transferred to a nylon membrane. 32HBV DNA synthesis was observed by autoradiography after hybridization with a specific probe for the full length of P-labeled HBV. Among the RI DNAs, partially double-stranded relaxed circular DNA (RC DNA) and doubled-stranded linear DNA (DL DNA) were labeled RC and DL, respectively.

[0294] Figure 26 is the result of an experiment on the inhibition of HBV proliferation in liver cancer cell lines by a compound according to Example 1 of the present invention.

[0295] In HepG2 and Huh7 cells in Fig. 26, panels 1 through 5 show Western blot results, panel 6 shows core particles formed by NAGE-immunoblotting, and the last panel shows Southern blotting to observe HBV RI DNA replication. HBV proliferation was demonstrated through HBc protein expression (4th panel), core particle formation (6th panel), and HBV RI DNA Southern blotting (last panel). Sirtuin 2 increased when HBV proliferated (lane 2) (3rd panels, lanes 2). Comparisons were made using AGK2, confirming that AGK2 inhibited SIRT2 and HBV proliferation (lanes 4). When HepG2 cells were treated with the compound, an AGK2-derived variant and SIRT2 inhibitor, at the indicated concentrations, there was no SIRT2 inhibition (3rd panel, lane 3), but HBV proliferation was inhibited at a concentration of 10 μM (lane 3). When Huh7 cells were treated with the compound at the labeled concentration (2.5 μM), there was no SIRT2 inhibition but HBV proliferation was inhibited (lane 3).

[0296] Referring to Fig. 26, it was confirmed that the compound according to Example 1 of the present invention inhibits HBV proliferation, and that it has superior efficacy in inhibiting HBV proliferation compared to AGK2. In addition, it was confirmed that it inhibits HBc(C) protein expression and the formation of core particles in HepG2 cells, and inhibits HBV DNA synthesis, and it was also confirmed that it inhibits HBV DNA synthesis in Huh7 cells.

[0297]

[0298] The foregoing description of the present invention is for illustrative purposes only, and those skilled in the art will understand that other specific forms can be easily modified without altering the technical concept or essential features of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive. For example, each component described as a single unit may be implemented in a distributed manner, and components described as distributed may likewise be implemented in a combined form.

[0299] The scope of the present invention is defined by the claims set forth below rather than by the detailed description above, and all modifications or variations derived from the meaning and scope of the claims and the concept of equivalents thereof should be interpreted as being included within the scope of the present invention.

Claims

1. Compound represented by the following chemical formula 1: [Chemical Formula 1] (In the above chemical formula 1, R is a linear or branched C1-C that can be substituted. 20 alkyl groups of, substitutable C3-C 20 cycloalkyl groups or C6-C that can be substituted 20 Arilgi, The above substitutions are oxygen, nitrogen, sulfur, hydroxyl, linear or branched C1-C6 alkyl groups, C3-C 20 cycloalkyl group of, C6-C 20 (Substituted by an aryl group, a halogen, or a combination thereof).

2. In Paragraph 1, A compound represented by the above chemical formula 1, comprising any one of the following compounds: ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; .

3. A step of preparing a compound represented by the following chemical formula 4 by reacting a compound represented by the following chemical formula 2 and a compound represented by the following chemical formula 3; A step of preparing a compound represented by the following chemical formula 6 by reacting a compound represented by the above chemical formula 4 with a compound represented by the following chemical formula 5; and A step of preparing a compound represented by Chemical Formula 1 of Claim 1 by substituting a hydroxyl group of a compound represented by Chemical Formula 6 above; including, Method for preparing a compound according to claim 1: [Chemical Formula 2] (In the above chemical formula 2, X 1 (is a halogen or B(OH)2) [Chemical Formula 3] (In the above chemical formula 3, X 2 is a halogen or B(OH)2) [Chemical Formula 4] [Chemical Formula 5] [Chemical Formula 6] .

4. In Paragraph 3, The above X 1 and X 2 One of them is a halogen and the other is B(OH)2, Method for manufacturing a compound according to claim 1.

5. A composition for inhibiting HBV proliferation, comprising a compound according to claim 1 as an active ingredient.

6. In Paragraph 5, The inhibition of HBc (Hepatitis B core) protein expression in HepG2 cells by a compound represented by the above chemical formula 1, Composition for inhibiting HBV proliferation.

7. In Paragraph 5, The formation of nucleocapsid in HepG2 cells is inhibited by the above compound, Composition for inhibiting HBV proliferation.

8. In Paragraph 5, HBV DNA synthesis in HepG2 cells is inhibited by the above compound, Composition for inhibiting HBV proliferation.

9. In Paragraph 5, HBV DNA synthesis in Huh7 cells is inhibited by the above compound, Composition for inhibiting HBV proliferation.

10. A pharmaceutical composition for preventing or treating HBV infection, comprising a composition for inhibiting HBV proliferation according to claim 5.

11. In Paragraph 10, The above pharmaceutical composition for the prevention or treatment of HBV infection further comprises a pharmaceutically acceptable carrier.

12. In Paragraph 11, A pharmaceutical composition for the prevention or treatment of HBV infection, wherein the carrier comprises a substance selected from the group consisting of lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, menthol, mineral oil, and combinations thereof.

13. In Paragraph 10, The above pharmaceutical composition for preventing or treating HBV infection further comprises a component selected from the group consisting of preservatives, solubilizers, stabilizers, humectants, sweeteners, coloring agents, flavoring agents, salts, buffers, antioxidants, lubricants, emulsifiers, suspending agents, preservatives, and combinations thereof.

14. In Paragraph 10, The above pharmaceutical composition for the prevention or treatment of HBV infection is administered by a method selected from the group consisting of intraperitoneal administration, oral administration, inhalation administration, intravenous administration, intramuscular administration, subcutaneous administration, dermal administration, intrauterine administration, tumor administration, rectal administration, and combinations thereof.

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