Novel compound

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 C1-C20 alkyl group, a substitutable C3-C20 cycloalkyl group, or a substitutable C6-C20 aryl group, and the substitution is carried out with oxygen, nitrogen, sulfur, hydroxy, a linear or branched C1-C6 alkyl group, a C3-C20 cycloalkyl group, an C6-C20 aryl group, a halogen, or a combination thereof.)

Description

New compound

[0001] The present invention relates to a novel compound.

[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 to develop 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 pharmaceutical composition for preventing or treating HBV infection is provided, comprising the above compound as an active ingredient.

[0014] 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.

[0015] 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:

[0016] [Chemical Formula 1]

[0017]

[0018] (In the above chemical formula 1,

[0019] 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,

[0020] 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).

[0021] 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:

[0022] ; ; ; ; ; ; ; ; ; ; ; .

[0023] Additionally, a second aspect of the present invention provides a method for preparing a compound according to a 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 chemical formula 6 to prepare a compound represented by the chemical formula 1 of claim 1.

[0024] [Chemical Formula 2]

[0025]

[0026] (In the above chemical formula 2, X 1 (is a halogen or B(OH)2)

[0027] [Chemical Formula 3]

[0028]

[0029] (In the above chemical formula 3, X 2 is a halogen or B(OH)2)

[0030] [Chemical Formula 4]

[0031]

[0032] [Chemical Formula 5]

[0033]

[0034] [Chemical Formula 6]

[0035] .

[0036] 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.

[0037] In addition, the third aspect of the present invention provides a pharmaceutical composition for preventing or treating HBV infection, comprising a compound according to the first aspect of the present invention as an active ingredient.

[0038] According to one embodiment of the present invention, the pharmaceutical composition may additionally comprise a pharmaceutically acceptable carrier, but is not limited thereto.

[0039] 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.

[0040] According to one embodiment of the present invention, the pharmaceutical composition 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.

[0041] According to one embodiment of the present invention, the pharmaceutical composition 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.

[0042] 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.

[0043] 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.

[0044] In addition, the inhibitory effect on HBV proliferation was confirmed compared to AGK2, and it was confirmed to effectively inhibit HBV proliferation in Huh7 cells. Specifically, it showed effects of inhibiting HBc(C) protein expression, inhibiting core particle (nucleocapsid or capsid) formation, and inhibiting HBV DNA synthesis.

[0045] In addition, viral replication can be effectively blocked by simultaneously inhibiting various stages of the HBV life cycle (protein expression, particle formation, DNA synthesis).

[0046] Furthermore, the compound according to the present invention was confirmed to inhibit the secretion of HBsAg and HBeAg in a concentration-dependent manner. This suggests that the compound of the present invention can effectively block not only viral replication but also the antigen secretion stage.

[0047] However, the effects obtainable from this invention are not limited to those described above, and other effects may exist.

[0048] Figure 1 is the NMR spectrum of a compound according to Example 1 of the present invention.

[0049] Figure 2 is the NMR spectrum of a compound according to Example 2 of the present invention.

[0050] Figure 3 is the NMR spectrum of a compound according to Example 3 of the present invention.

[0051] Figure 4 is the NMR spectrum of a compound according to Example 4 of the present invention.

[0052] Figure 5 is the NMR spectrum of a compound according to Example 5 of the present invention.

[0053] Figure 6 is the NMR spectrum of a compound according to Example 6 of the present invention.

[0054] Figure 7 is the NMR spectrum of a compound according to Example 7 of the present invention.

[0055] Figure 8 is the NMR spectrum of a compound according to Example 8 of the present invention.

[0056] Figure 9 is the NMR spectrum of a compound according to Example 9 of the present invention.

[0057] Figure 10 is the NMR spectrum of a compound according to Example 10 of the present invention.

[0058] Figure 11 is the NMR spectrum of a compound according to Example 11 of the present invention.

[0059] Figure 12 is the NMR spectrum of a compound according to Example 12 of the present invention.

[0060] Figure 13 is the result of a cytotoxicity test according to an experimental example of the present invention.

[0061] Figure 14 is the result of an HBV proliferation inhibition experiment according to an experimental example of the present invention.

[0062] Figure 15 is the ELISA result of a compound according to Example 1 (ACTC-004) of the present invention.

[0063] Figure 16 is the ELISA result of a compound according to Example 3 (ACTC-024) of the present invention.

[0064] Figure 17 is the ELISA result of a compound according to Example 4 (ACTC-025) of the present invention.

[0065] Figure 18 is the ELISA result of a compound according to Example 9 (ACTC-047) of the present invention.

[0066] Figure 19 is the ELISA result of a compound according to Example 11 (ACTC-054) of the present invention.

[0067] Embodiments of the present invention are described below with reference to the attached drawings so that those skilled in the art can easily implement them. 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.

[0068] 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.

[0069] 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.

[0070] 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.

[0071] 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 present invention. Furthermore, throughout this specification, “a step of” or “a step of” does not mean “a step for”.

[0072] 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.

[0073] Throughout the entire specification, the description "A and / or B" means "A, B, or A and B".

[0074] Hereinafter, the novel compounds of the present invention 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.

[0075]

[0076] 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:

[0077] [Chemical Formula 1]

[0078]

[0079] (In the above chemical formula 1,

[0080] 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,

[0081] 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).

[0082] 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.

[0083] In addition, the inhibitory effect on HBV proliferation was confirmed compared to AGK2, and it was confirmed to effectively inhibit HBV proliferation in Huh7 cells. Specifically, it showed effects of inhibiting HBc(C) protein expression, inhibiting core particle (nucleocapsid or capsid) formation, and inhibiting HBV DNA synthesis.

[0084] In addition, viral replication can be effectively blocked by simultaneously inhibiting various stages of the HBV life cycle (protein expression, particle formation, DNA synthesis).

[0085] 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:

[0086] ; ; ; ; ; ; ; ; ; ; ; .

[0087]

[0088] Additionally, a second aspect of the present invention provides a method for preparing a compound according to a 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 chemical formula 6 to prepare a compound represented by the chemical formula 1 of claim 1.

[0089] [Chemical Formula 2]

[0090]

[0091] (In the above chemical formula 2, X 1 (is a halogen or B(OH)2)

[0092] [Chemical Formula 3]

[0093]

[0094] (In the above chemical formula 3, X 2 is a halogen or B(OH)2)

[0095] [Chemical Formula 4]

[0096]

[0097] [Chemical Formula 5]

[0098]

[0099] [Chemical Formula 6]

[0100] .

[0101] 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.

[0102] 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.

[0103] 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.

[0104]

[0105] In addition, the third aspect of the present invention provides a pharmaceutical composition for preventing or treating HBV infection, comprising a compound according to the first aspect of the present invention as an active ingredient.

[0106] Regarding the pharmaceutical composition 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 are 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.

[0107] According to one embodiment of the present invention, the pharmaceutical composition may additionally comprise a pharmaceutically acceptable carrier, but is not limited thereto.

[0108] 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.

[0109] According to one embodiment of the present invention, the pharmaceutical composition 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.

[0110] According to one embodiment of the present invention, the pharmaceutical composition 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.

[0111] 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.

[0112]

[0113] [Example 1] ACT-C-004

[0114] (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)-N-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)acrylamide (JYPf70)

[0115] 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 1,4-benzodioxane-6-amine (35.9 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 obtain the target compound as a yellow powder (10.1 mg, 9.4%).

[0116]

[0117] Figure 1 is the NMR spectrum of a compound according to Example 1 of the present invention.

[0118] 1H NMR (600 MHz, CDCl3) δ 8.15-8.17 (m, 2H), 7.88 (s, 1H), 7.47 (d,J = 3.4 Hz, 1H), 7.42 (d,J = 8.3 Hz, 1H), 7.29 (td,J = 8.8, 2.3 Hz, 3H), 6.99 (dd,J = 8.6, 2.4 Hz, 1H), 6.87 (d,J = 8.3 Hz, 1H), 4.26-4.29 (m, 4H)

[0119] [Example 2] ACT-C-005

[0120] (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)-N-(4-(dimethylamino)phenyl)acrylamide (JYPf80)

[0121] 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 N,N-dimethyl-1,4-phenylenediamine (39.8 mg, 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 the target compound as a yellow powder (10.1 mg, 9.7%).

[0122]

[0123] Figure 2 is the NMR spectrum of a compound according to Example 2 of the present invention.

[0124] 1H NMR (600 MHz, CDCl3) δ 8.17 (d,J = 2.1 Hz, 1H), 8.14 (s, 1H), 7.89 (s, 1H), 7.47 (t,J = 4.5 Hz, 3H), 7.42 (d,J = 8.3 Hz, 1H), 7.29 (dd,J) = 8.6, 2.4 Hz, 1H), 7.24 (d,J = 3.4 Hz, 1H), 6.73-6.74 (m, 2H), 2.96 (s, 6H)

[0125] [Example 3] ACT-C-024

[0126] (E)-tert-butyl 4-(2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylamido)piperidine-1-carboxylate (JYPg05)

[0127] 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-amino-1-tert-butoxycarbonylpiperidine (117 mg, 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 the target compound as a pale yellow powder. (92 mg, 38.5%)

[0128]

[0129] Figure 3 is the NMR spectrum of a compound according to Example 3 of the present invention.

[0130] 1H NMR (600 MHz, CDCl3) δ 8.14 (d,J = 2.8 Hz, 1H), 7.64 (s, 1H), 7.43 (d,J = 3.4 Hz, 1H), 7.40 (d,J = 9.0 Hz, 1H), 7.27 (d,J = 2.8 Hz, 1H), 7.18 (d,J = 4.1 Hz, 2H), 4.11 (m, 8H), 2.08 (m, 2H), 1.44 (s, 9H)

[0131] [Example 4] ACT-C-025

[0132] (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)-N-(4-hydroxycyclohexyl)acrylamide (JYPg43)

[0133] 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 aminocyclohexanol (67.3 mg, 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. (46.0 mg, 23.4%)

[0134]

[0135] Figure 4 is the NMR spectrum of a compound according to Example 4 of the present invention.

[0136] 1H NMR (600 MHz, CD3OD) δ 8.08 (d,J = 2.8 Hz, 1H), 7.98 (s, 1H), 7.42 (d,J = 3.4 Hz, 1H), 7.38 (d,J = 8.3 Hz, 1H), 7.25 (dd,J = 8.6, 2.4 Hz, 1H), 7.22 (d,J = 4.1 Hz, 1H), 3.78-3.82 (m, 1H), 3.51-3.57 (m, 1H), 1.98 (m, 4H), 1.27-1.42 (m, 4H)

[0137] [Example 5] ACT-C-038

[0138] (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)-N-(4-(trifluoromethyl)phenyl)acrylamide (JYPg99)

[0139] 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-trifluoromethylaniline (73.4 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 dried with Na2SO4, filtered, and concentrated. The residue was purified by silica gel column chromatography to obtain the target compound. (55.0 mg, 28.4%)

[0140]

[0141] Figure 5 is the NMR spectrum of a compound according to Example 5 of the present invention.

[0142] 1H NMR (600 MHz, CDCl3) δ 8.22-8.09 (m, 3H), 7.82-7.71 (d,J = 8.3 Hz, 2H), 7.71-7.60 (d,J = 8.3 Hz, 2H), 7.55-7.46 (d,J = 4.1 Hz, 1H), 7.46-7.38 (d,J = 8.3 Hz, 1H), 7.36-7.29 (d,J = 5.5 Hz, 2H).

[0143] [Example 6] ACT-C-040

[0144] (E)-N-((3s,5s,7s)-adamantan-1-yl)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylamide (JYPh03)

[0145] A solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (150 mg, 0.487 mmol) dissolved in thionyl chloride (20 mL) was stirred at reflux temperature for 3 hours. After evaporating the solution under reduced pressure, the residue was dissolved in dichloromethane and added to a dichloromethane solution of adamanthylamine (88.4 mg, 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 added to 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 (41.0 mg, 19.1%).

[0146]

[0147] Figure 6 is the NMR spectrum of a compound according to Example 6 of the present invention.

[0148] 1H NMR (600 MHz, CDCl3) δ 8.19-8.10 (d,J = 2.1 Hz, 1H), 7.99 (s, 1H), 7.48-7.35 (m, 2H), 7.32-7.27 (d,J = 2.1 Hz, 1H), 7.22-7.12 (d,J = 3.4 Hz, 1H), 6.05 (s, 1H), 2.19-2.03 (m, 9H), 1.79-1.65 (m, 6H).

[0149] [Example 7] ACT-C-041

[0150] (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)-N-(pyridine-2-yl)acrylamide (JYPh05)

[0151] A solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (150 mg, 0.487 mmol) dissolved in thionyl chloride (20 mL) was stirred at reflux temperature for 3 hours. After evaporating the solution under reduced pressure, the residue was dissolved in dichloromethane and added to a dichloromethane solution of 2-aminopyridine (55.0 mg, 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 added to 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 (9.3 mg, 5.0%).

[0152]

[0153] Figure 7 is the NMR spectrum of a compound according to Example 7 of the present invention.

[0154] 1H NMR (600 MHz, CDCl3) δ 8.66 (s, 1H), 8.41-8.32 (d,J = 4.1 Hz, 1H), 8.32-8.22 (d,J = 8.3 Hz, 1H), 8.19 (s, 1H), 8.17-8.12 (d,J = 2.8 Hz, 1H), 7.82-7.71 (m, 1H), 7.53-7.46 (d,J = 4.1 Hz, 1H), 7.46-7.38 (d,J = 8.3 Hz, 1H), 7.38-7.28 (m, 2H), 7.18-7.07 (dd, J = 6.9, 4.8 Hz, 1H).

[0155] [Example 8] ACT-C-042

[0156] (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)-N-(quinoline-2-yl)acrylamide (JYPh27)

[0157] A solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (150 mg, 0.487 mmol) dissolved in thionyl chloride (20 mL) was stirred at reflux temperature for 3 hours. After evaporating the solution under reduced pressure, the residue was dissolved in dichloromethane and added to a dichloromethane solution of 2-aminoquinoline (84.2 mg, 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 added to 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 (18.2 mg, 8.6%).

[0158]

[0159] Figure 8 is the NMR spectrum of a compound according to Example 8 of the present invention.

[0160] 1H NMR (600 MHz, CDCl3) δ 8.90 (s, 1H), 8.45 (d,J = 6.9 Hz, 1H), 8.24 (d,J = 4.1 Hz, 2H), 8.19 (d,J = 2.8 Hz, 1H), 7.91 (d,J = 7.6 Hz, 1H), 7.82 (d,J = 7.6 Hz, 1H), 7.73-7.70 (m, 1H), 7.51-7.49 (m, 2H), 7.44-7.43 (m, 1H), 7.36 (s, 1H), 7.31 (dd,J = 8.6, 2.4 Hz, 1H).

[0161] [Example 9] ACT-C-047

[0162] (E)-2-cyano-N-cycloheptyl-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylamide (YJb46)

[0163] 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 cycloheptylamine (74 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 added to 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 11:89) to obtain the target compound (79 mg, 40%).

[0164]

[0165] Figure 9 is the NMR spectrum of a compound according to Example 9 of the present invention.

[0166] 1H NMR (600 MHz, CDCl3) δ 8.13 (d,J = 2.5 Hz, 1H), 8.03 (s, 1H), 7.42 (d,J = 3.8 Hz, 1H), 7.39 (d,J = 8.5 Hz, 1H), 7.27-7.25 (m, 1H), 7.18 (d,J = 3.8 Hz, 1H), 6.27 (d,J = 8.1 Hz, 1H), 4.08 (dt,J = 9.0, 4.5 Hz, 1H), 2.03-1.95 (m, 2H), 1.69-1.60 (m, 4H), 1.56-1.50 (m, 6H).

[0167] [Example 10] ACT-C-052

[0168] (E)-N-(tert-butyl)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylamide (YJb60)

[0169] 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 tert-butylamine (62 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 (36 mg, 20%).

[0170]

[0171] Figure 10 is the NMR spectrum of a compound according to Example 10 of the present invention.

[0172] 1 H NMR (600 MHz, CDCl3) δ 8.15 (d,J = 2.5 Hz, 1H), 8.01 (s, 1H), 7.44 (d,J = 3.8 Hz, 1H), 7.41 (d,J = 8.5 Hz, 1H), 7.28-7.27 (m, 1H), 7.17 (d,J = 3.8 Hz, 1H), 6.19 (s, 1H), 1.45 (s, 9H).

[0173] [Example 11] ACT-C-054

[0174] tert-butyl (E)-(4-(2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylamido)phenyl)carbamate (YJb25)

[0175] Thionyl chloride (15 mL) 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 N-(tert-butoxycarbonyl)-1,4-phenylenediamine (122 mg, 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 14 hours. The product was purified by flash column chromatography (EA / Hex; 5:95 to 15:85) to obtain the target compound (9 mg, 4%).

[0176]

[0177] Figure 11 is the NMR spectrum of a compound according to Example 11 of the present invention.

[0178] 1H NMR (600 MHz, CDCl3) δ 8.15 (d,J = 2.5 Hz, 1H), 8.14 (s, 1H), 7.95 (s, 1H), 7.54 (d,9.0 Hz, 2H), 7.46 (d,J = 3.7 Hz,1H), 7.41 (d,J = 8.5 Hz, 1H), 7.38 (d,J = 8.4 Hz, 2H), 7.28 (dd,J = 8.5, 2.5 Hz, 1H), 6.47 (s, 1H), 1.51 (s, 9H).

[0179] [Example 12] ACT-C-063

[0180] (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)-N-(quinoline-6-yl)acrylamide (JYPg09)

[0181] A solution of (E)-2-cyano-3-(5-(2,5-dichlorophenyl)furan-2-yl)acrylic acid (150 mg, 0.487 mmol) dissolved in thionyl chloride (20 mL) was stirred at reflux temperature for 3 hours. After evaporating the solution under reduced pressure, the residue was dissolved in dichloromethane and added to a dichloromethane solution of 6-aminoquinoline (84.2 mg, 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 added to 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 (15.3 mg, 7.2%).

[0182]

[0183] Figure 12 is the NMR spectrum of a compound according to Example 12 of the present invention.

[0184] 1H NMR (600 MHz, CDCl3) δ 10.66 (s, 1H), 8.84-8.83 (m, 1H), 8.41 (d,J = 2.1 Hz, 1H), 8.35 (d,J = 8.3 Hz, 1H), 8.25 (s, 1H), 8.17 (d,J = 2.8 Hz, 1H), 8.02-8.05 (m, 1H), 7.97 (dd,J = 9.3, 2.4 Hz, 1H), 7.70 (d,J = 9.0 Hz, 1H), 7.63 (d,J = 4.1 Hz, 1H), 7.51-7.58 (m, 3H).

[0185]

[0186] [Experimental Example 1] MTS assay: Cytotoxicity test

[0187] Experiments were performed to measure the cytotoxicity of the compounds according to Examples 1 to 12 (excluding Example 9).

[0188] 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 at 37°C under 5% CO2. The following day, the cells were treated with compounds at various concentrations (1, 5, 10, 50, 100 μM) and cultured for 48 hours (as in Examples 1-4). For Examples 5-12, the cells were cultured for a total of 72 hours after compound treatment. 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, the absorbance was measured at a plate reader OD=490 nm.

[0189] Figure 13 is the result of a cytotoxicity test according to an experimental example of the present invention.

[0190] Referring to FIG. 13, none of the compounds according to Examples 1 to 12 showed cytotoxicity in HepG2 liver cancer cell lines at 1, 5, 10, 50, and 100 μM, and CC 50 I couldn't calculate the value.

[0191]

[0192] [Experimental Example 2] Results of HBV proliferation inhibition experiment

[0193] An experiment was performed to confirm the HBV proliferation inhibitory effect of the compound according to Example 1.

[0194] Figure 14 is the result of an HBV proliferation inhibition experiment according to an experimental example of the present invention.

[0195] Referring to Figure 14, it was confirmed that at 5 μM, SIRT2 decreased, and AGK2 inhibited SIRT2 to a similar degree as at 2.5 μM (3rd panel). In other words, it was confirmed that the compound did not lose its SIRT2 inhibitory function.

[0196] In addition, a decrease in HBc expression (4th panel), a decrease in core particle formation (6th panel), and a decrease in HBV DNA synthesis (last panel; lanes 3,4) were observed.

[0197]

[0198] [Experimental Example 3] Evaluation of the HBsAg and HBeAg secretion inhibitory effect of compounds using ELISA

[0199] Each compound was treated simultaneously with transfection of 1.3mer HBV WT into Huh7 cells. Example 1 (ATCT-004), Example 3 (ATCT-0024), Example 4 (ATCT-0025), Example 9 (ATCT-0047), and Example 11 (ATCT-0054) were treated at concentrations of 2.5 μM and 5 μM, respectively, and AGK2 2.5 μM was used as a control.

[0200]

[0201] After 72 hours of transfection, the culture supernatant was collected, and ELISA was performed to confirm the inhibitory effects of the compounds according to Example 1 (ACTC-004), Example 3 (ACTC-024), Example 4 (ACTC-025), Example 9 (ACTC-047), and Example 11 (ACTC-054) on HBsAg and HBeAg secretion. The results were analyzed using the WANTAI HBsAg ELISA kit (WB-2296) and the WANTAI HBeAg ELISA kit (WB-2496).

[0202] FIGS. 15 to 19 are ELISA results of compounds according to Example 1 (ACTC-004), Example 3 (ACTC-024), Example 4 (ACTC-025), Example 9 (ACTC-047) and Example 11 (ACTC-054) of the present invention, respectively.

[0203] Referring to Fig. 15, the HBsAg reduction effect by Example 1 (ATCT-004) was weaker than that of AGK2 and showed a lower reduction rate than the HBV proliferation reduction results shown in Fig. 14. In Fig. 16, Example 3 (ATCT-0024) reduced HBsAg secretion to a level similar to that of AGK2 at the same concentration of 2.5 μM, and showed a greater reduction effect than 2.5 μM AGK2 at a concentration of 5 μM. In Fig. 17, Example 4 (ATCT-0025) reduced HBsAg to a level similar to that of AGK2 at the same concentration of 2.5 μM, and showed a greater reduction in HBsAg secretion than 2.5 μM AGK2 at a concentration of 5 μM.

[0204] In Fig. 18, Example 9 (ATCT-0047) showed a greater HBsAg reduction effect than AGK2 at the same concentration at a concentration of 2.5 μM. In Fig. 19, Example 11 (ATCT-0054) showed a significantly superior HBsAg secretion inhibitory effect than the control AGK2 at the same concentration at a concentration of 2.5 μM, and at a concentration of 5 μM, although the degree of reduction was somewhat lower than at 2.5 μM, it still maintained a significantly superior inhibitory effect than AGK2.

[0205] Referring to the right-hand diagrams of Figures 15 to 19, no change was observed in the HBeAg secretion ELISA results of Huh7 cells transfected with AGK2 in all experimental groups.

[0206]

[0207] 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.

[0208] 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 pharmaceutical composition for preventing or treating HBV infection, comprising a compound according to claim 1 as an active ingredient.

6. In Paragraph 5, The above pharmaceutical composition comprises a pharmaceutically acceptable carrier.

7. In Paragraph 6, A pharmaceutical composition comprising the carrier 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.

8. In Paragraph 5, The above pharmaceutical composition 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.

9. In Paragraph 5, The above pharmaceutical composition 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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