NOVEL COMPOUND FOR INHIBITING HISTONE ACETYLTRANSFERASE P300 AND ANTIFIBROTIC COMPOSITION COMPRISING SAME

Abstract

The present invention relates to a novel compound that enables formation of an additional hydrogen bond with a specific amino acid position in histone acetyltransferase (HAT) p300 through the structural analysis of the HAT p300. The novel compound of the present invention has a significantly excellent inhibitory effect on the activity of HAT p300 and can be used very effectively in the prevention, amelioration or treatment of diseases associated with the activation of HAT p300, such as fibrosis.

Claims

1. A compound selected from a compound represented by the following Formula 1, and a pharmaceutically acceptable salt, an optical isomer, a hydrate, and a solvate thereof: ##STR00010## wherein: m and n are each independently an integer ranging from 1 to 4; R.sub.1 is a substituent selected from the group consisting of a substituted or unsubstituted C.sub.3-C.sub.40 cycloalkyl group, a heterocycloalkyl group having 5 to 7 nuclear atoms, a C.sub.6-C.sub.14 aryl group, and a heteroaryl group having 5 to 14 nuclear atoms; p is an integer ranging from 1 to 3, and q is an integer ranging from 0 to 3, provided that p+q is not greater than 4; r is an integer ranging from 0 to 5; R.sub.2 is a halogen, wherein when R.sub.2 is present in a plural number, they are the same or different from each other; and R.sub.3 is a substituent selected from the group consisting of a halogen, a cyano group (—CN), a C.sub.1-C.sub.5 alkylcarbonyl group, and a carbamoyl group (—C(═O)—(NH.sub.2)), wherein when R.sub.3 is present in a plural number, they are the same or different from each other.

2. The compound of claim 1, wherein, in Formula 1, m and n are each independently an integer of 1 or 2; R.sub.1 is a substituted or unsubstituted C.sub.3-C.sub.40 cycloalkyl group, a heterocycloalkyl group having 5 to 7 nuclear atoms, a C.sub.6-C.sub.14 aryl group, and a heteroaryl group having 5 to 14 nuclear atoms; p is an integer of 1 or 2; q is an integer of 0 or 1; and r is an integer of 1 or 2.

3. The compound of claim 2, wherein R.sub.1 is a C.sub.6-C.sub.14 aryl group; the aryl group of R.sub.1 is unsubstituted or substituted with at least one —(COO(R.sub.4)); and R.sub.4 is hydrogen or a C.sub.1-C.sub.4 alkyl group.

4. The compound of claim 1, wherein m and n are each independently an integer of 1 or 2; r is an integer of 1; R.sub.1 is a substituent selected from a heterocycloalkyl group having 5 to 7 nuclear atoms or a C.sub.6-C.sub.8 aryl group; R.sub.2 is a halogen; R.sub.3 is a halogen or a cyano group; the C.sub.6-C.sub.8 aryl group of R.sub.1 is substituted with —(COO(R.sub.4)); and R.sub.4 is hydrogen or a C.sub.1-C.sub.4 alkyl group.

5. The compound of claim 1, wherein the compound comprises any one selected from the group consisting of the following compounds: ##STR00011## ##STR00012## ##STR00013##

6. A method for inhibiting histone acetyltransferase p300, comprising: administering a compound selected from a compound represented by the following Formula 1, and a pharmaceutically acceptable salt, an optical isomer, a hydrate, and a solvate thereof: ##STR00014## wherein: m and n are each independently an integer ranging from 1 to 4; R.sub.1 is a substituent selected from the group consisting of a substituted or unsubstituted C.sub.3-C.sub.40 cycloalkyl group, a heterocycloalkyl group having 5 to 7 nuclear atoms, a C.sub.6-C.sub.14 aryl group, and a heteroaryl group having 5 to 14 nuclear atoms; p is an integer ranging from 1 to 3, and q is an integer ranging from 0 to 3, provided that p+q is not greater than 4; r is an integer ranging from 0 to 5; R.sub.2 is a halogen, wherein when R.sub.2 is present in a plural number, they are the same or different from each other; and R.sub.3 is a substituent selected from the group consisting of a halogen, a cyano group (—CN), a C.sub.1-C.sub.5 alkylcarbonyl group, and a carbamoyl group (—C(═O)—(NH.sub.2)), wherein when R.sub.3 is present in a plural number, they are the same or different from each other.

7. A pharmaceutical composition for preventing or treating a histone acetyltransferase p300-associated disease, comprising, as an active ingredient, a compound selected from a compound represented by the following Formula 1, and a pharmaceutically acceptable salt, an optical isomer, a hydrate, and a solvate thereof: ##STR00015## wherein: m and n are each independently an integer ranging from 1 to 4; R.sub.1 is a substituent selected from the group consisting of a substituted or unsubstituted C.sub.3-C.sub.40 cycloalkyl group, a heterocycloalkyl group having 5 to 7 nuclear atoms, a C.sub.6-C.sub.14 aryl group, and a heteroaryl group having 5 to 14 nuclear atoms; p is an integer ranging from 1 to 3, and q is an integer ranging from 0 to 3, provided that p+q is not greater than 4; r is an integer ranging from 0 to 5; R.sub.2 is a halogen, wherein when R.sub.2 is present in a plural number, they are the same or different from each other; and R.sub.3 is a substituent selected from the group consisting of a halogen, a cyano group (—CN), a C.sub.1-C.sub.5 alkylcarbonyl group, and a carbamoyl group (—C(═O)—(NH.sub.2)), wherein when R.sub.3 is present in a plural number, they are the same or different from each other.

8. The pharmaceutical composition of claim 7, wherein the histone acetyltransferase p300-associated disease is fibrosis.

9. The pharmaceutical composition of claim 8, wherein the fibrosis comprises one or more selected from the group consisting of pulmonary fibrosis, uterine myoma, myelofibrosis, liver fibrosis, heart fibrosis, multiple sclerosis, kidney fibrosis, cystic fibrosis, neutropenia, skeletal muscle fibrosis, scleroderma, dermatomyositis, mediastinal fibrosis, and splenic fibrosis caused by sickle-cell anemia.

10. The pharmaceutical composition of claim 9, wherein the pulmonary fibrosis one or more selected from the group consisting of idiopathic pulmonary fibrosis, nonspecific interstitial pneumonia, acute interstitial pneumonia, cryptogenic organizing pneumonia, a respiratory bronchiolitis-associated interstitial lung disease, desquamative interstitial pneumonia, lymphoid interstitial pneumonia, interstitial pulmonary fibrosis, and diffuse pulmonary fibrosis.

11. A method for preventing or ameliorating a histone acetyltransferase p300-associated disease, comprising administering or taking a food composition comprising a compound according to claim 1 to a subject in need thereof ##STR00016##

12. A method for preventing or ameliorating a histone acetyltransferase p300-associated disease, comprising administering or applying a cosmetic composition comprising a compound according to claim 1 to a subject in need thereof ##STR00017##

13. A method for preventing or treating a histone acetyltransferase p300-associated disease, comprising: administering a compound according to claim 1 to a target subject ##STR00018##

14. The method of claim 13, wherein the histone acetyltransferase p300-associated disease is fibrosis.

Description

DESCRIPTION OF DRAWINGS

[0079] FIGS. 1 to 3 show the results of confirming, through immunohistochemical staining, expression levels of histone acetyltransferase (HAT) proteins, i.e., p300 (histone acetyltransferase p300; FIG. 1), GCN5 (histone acetyltransferase GCN5; FIG. 2), and PCAF (P300/CBP-associated factor; FIG. 3), in the tissue from a patient with idiopathic pulmonary fibrosis according to one embodiment of the present invention.

[0080] FIG. 4 shows the results of confirming the degree of inhibition of HAT activity of Candidates 1 to 67, HAT-24, HAT-26, and HAT-28 that are PCAF inhibitors according to one embodiment of the present invention.

[0081] FIG. 5 shows the results of confirming the degree of inhibition of HAT activity of Candidates 1 to 14 according to one embodiment of the present invention.

[0082] FIG. 6 shows a Lys-CoA molecule model of a HAT p300 domain and a substrate inhibitor using a molecular docking simulation according to one embodiment of the present invention.

[0083] FIG. 7 shows the results of analyzing main residues that participate in an intermolecular bond between the HAT p300 domain and the substrate inhibitor using the molecular docking simulation according to one embodiment of the present invention.

[0084] FIG. 8 shows the results of analyzing main residues that participate in a bond between Candidate 12 and the HAT p300 domain using the molecular docking simulation according to one embodiment of the present invention.

[0085] FIG. 9 shows the results of confirming the degree of inhibition of histone acetyltransferase activity of Synthesis Examples 1 to 19 (A 1 to A 19) according to one embodiment of the present invention.

BEST MODE

[0086] According to one embodiment of the present invention, there is provided a compound selected from a compound represented by the following Formula 1, and a pharmaceutically acceptable salt, an optical isomer, a hydrate, and a solvate thereof:

##STR00007##

[0087] wherein m and n are each independently an integer ranging from 1 to 4; R.sub.1 is a substituent selected from the group consisting of a substituted or unsubstituted C.sub.3-C.sub.40 cycloalkyl group, a heterocycloalkyl group having 5 to 7 nuclear atoms, a C.sub.6-C.sub.14 aryl group, and a heteroaryl group having 5 to 14 nuclear atoms; p is an integer ranging from 1 to 3, q is an integer ranging from 0 to 3, provided that p+q is not greater than 4; r is an integer ranging from 0 to 5; R.sub.2 is a halogen, wherein when R.sub.2 is present in a plural number, they are the same or different from each other; and R.sub.3 is a substituent selected from the group consisting of a halogen, a cyano group (—CN), a C.sub.1-C.sub.5 alkylcarbonyl group, and a carbamoyl group (—C(═O)—(NH.sub.2)), wherein when R.sub.3 is present in a plural number, they are the same or different from each other.

[0088] According to another embodiment of the present invention, there is provided a composition for preventing, ameliorating or treating a HAT p300-associated disease, which includes, as an active ingredient, a compound selected from a compound represented by Formula 1, and a pharmaceutically acceptable salt, an optical isomer, a hydrate, and a solvate thereof.

[0089] According to still another embodiment of the present invention, there is provided a method of preventing, ameliorating or treating a HAT p300-associated disease, which includes, as an active ingredient, a compound selected from a compound represented by Formula 1, and a pharmaceutically acceptable salt, an optical isomer, a hydrate, and a solvate thereof.

MODE FOR INVENTION

[0090] Hereinafter, the present invention will be described in detail with reference to the following examples. It will be apparent to those skilled in the art that the following examples are merely provided to exemplify the present invention, and are not intended to limit the scope of the present invention without departing from the scope of the present invention.

EXAMPLES

[Example 1] Confirmation of Protein Expression Level in Tissue of Fibrosis Patient

[0091] Tissues obtained from a patient with idiopathic pulmonary fibrosis or a normal person were fixed in 10% formalin, and embedded in paraffin, and a 7 m-thick section was attached to a slide. Thereafter, the section was deparaffinized using xylene, and treated with a high concentration to a low concentration of ethanol. Then, immunostaining was performed using antibodies specific to HAT p300 (histone acetyltransferase p300), GCN5 (histone acetyltransferase GCN5), and PCAF (P300/CBP-associated factor), and an expression level of each protein was measured using an optical microscope. The results are shown in FIGS. 1 to 3.

[0092] As shown in FIGS. 1 to 3, it was confirmed that, among the histone acetyltransferases (hereinafter referred to as “HAT”) p300, GCN5, and PCAF, the expression of p300 increased in the tissue from the patient with idiopathic pulmonary fibrosis, compared to the tissue obtained from the normal person.

[0093] Based on the results, it can be seen that the fibrosis is able to be effectively treated when the expression or function of p300 is specifically inhibited because especially p300 among the histone acetyltransferases is present at a high level in the fibrosis such as idiopathic pulmonary fibrosis, compared to the normal tissue.

[Example 2] Screening of p300 Activity Inhibitors

[0094] [2-1] Primary Screening

[0095] Candidates 1 to 67, which are HAT inhibitors manufactured based on the PCAF structure, and HAT-24, HAT-26, and HAT-28 were diluted to 100 μM, and the degree of inhibition of HAT activity of p300 (inhibitory activity) was then determined using a kit for measuring HAT activity (Biovision, Cat No. K332, U.S.A) according to the method provided by the manufacturer. The results are shown in FIG. 4. Here, the tissue was treated with C646 (HAT inhibitor) as a positive control.

[0096] As shown in FIG. 4, 80% of the HAT activity was inhibited by Candidates 1 to 14, but Candidate 15 to 67, HAT-25, HAT-26, and HAT-28 had a HAT activity inhibitory effect of only 20% to 50%.

[0097] [2-2] Secondary Screening

[0098] Candidates 1 to 14 having a good HAT activity inhibitory effect in Section [2-1] were diluted to concentrations of 0.5 μM, 1 μM, 10 μM, and 100 μM. Thereafter, the degree of inhibition of HAT activity was determined in the same manner as in Section [2-1]. The results are shown in FIG. 5 and Table 1.

TABLE-US-00001 TABLE 1 Candidate 1 2 3 4 5 6 7 8 9 10 11 12 13 14 IC.sub.50 (μM) 50.84 44.09 28.9 12.12 32.4 10.14 27.65 35.52 9.01 13.12 41.67 0.953 32.84 35.82

[0099] As shown in FIG. 5 and Table 1, it was confirmed that among Candidates 1 to 14, Candidate 12 (HAT-12) had an IC.sub.50 of 0.953. Here, Candidate 12 is as described in the following Formula 3.

##STR00008##

[Example 3] Structural Analysis of Candidate 12

[0100] To obtain information on the structure-activity correlation of a candidate through a molecular docking simulation, a molecule model was established using a co-crystal structure (pdb: 3biy) of a HAT p300 domain and its substrate inhibitor (Lys-CoA). The Lys-CoA was finally re-docked into the molecule model thus established, and the co-crystal structures were compared. As a result, it was confirmed that the candidate had a similar binding pattern. From the results, the accuracy of the docking results was evaluated (FIG. 6), the main residues participating in the binding between the HAT p300 domain and the Lys-CoA were analyzed (FIG. 7), and a molecular docking simulation was performed with Candidate 12 (HAT-12) (FIG. 8).

[0101] As shown in FIG. 6, it was confirmed that there was a high similarity between a predicted binding pattern of the HAT p300 domain-ligand (Lys-CoA) and the co-crystal structure.

[0102] As shown in FIG. 7, it was confirmed that, when the main residues participating in the binding between the Lys-CoA and the HAT p300 domain were analyzed, hydrogen bonds were observed at R1410, T1411, W1466, Y1467, L1398, S1400, 11457, W1436, and Y1397. Among these, the interactions with R1410, T1411, W1466, and Y1467 were important for drug inhibitory activity. Especially, it can be seen that W1466 played a key role in drug inhibitory activity (see, for example, Erin M. Bowers et al., Virtual Ligand Screening of the p300/CBP Histone Acetyltransferase: Identification of a Selective Small Molecule Inhibitor, Chem Biol. 2010 May 28; 17(5): 471-82).

[0103] As shown in FIG. 8, it was confirmed that, when the main residues participating in the binding between Candidate 12 and the HAT p300 domain were analyzed, the interactions with R1410, T1411, W1466, and Y1467 were not observed, but hydrogen bonds were formed with L1398 and S1400.

[0104] From the results, it can be expected that L1398 and S1400 also played an important role in drug inhibitory activity. Also, it can be seen that when the additional hydrogen bonding with R1410, T1411, W1466, and Y1467 was hindered, HAT p300 inhibitory activity was further enhanced.

[0105] Based on these results, Synthesis Examples 1 to 19 having the above-described characteristics were prepared in order to further enhance HAT p300 inhibitory activity, as follows.

[Synthesis Examples 1 to 19] Preparation of Novel Compounds for Inhibiting Histone Acetyltransferase p300

[0106] ##STR00009##

[Synthesis Method 1] Compounds 1 to 17

[0107] Substituted 4-hydroxybenzaldehyde (1 equivalent) and its corresponding substituted benzyl chloride (1 equivalent), and K.sub.2CO.sub.3 (1 equivalent) were cultured in a DMF solvent at 80° C. for an hour. The reaction mixture was cooled to room temperature, and then extracted using ethyl acetate. The extract was washed with water, NaHCO.sub.3, and brine, and then dried over MgSO.sub.4. Thereafter, the solvent was completely removed under reduced pressure, and silica gel chromatography was then performed to obtain Compounds 1 to 17 as O-benzylated compounds (see Table 2).

TABLE-US-00002 TABLE 2 Compound R R.sub.1 R.sub.2 1 3-chlorophenyl OCH.sub.3 Cl 2 3-chlorophenyl OCH.sub.3 Br 3 3-chlorophenyl OCH.sub.3 I 4 3-ethoxycarbonylphenyl OCH.sub.3 Cl 5 3-ethoxycarbonylphenyl OCH.sub.3 Br 6 3-ethoxycarbonylphenyl OCH.sub.3 I 7 3-cyanophenyl OCH.sub.3 Cl 8 3-cyanophenyl OCH.sub.3 Br 9 3-cyanophenyl OCH.sub.3 I 10 3-fluorophenyl OCH.sub.3 Cl 11 3-chlorophenyl OCH.sub.3 H 12 4-carbamoylphenyl OCH.sub.3 Cl 13 4-carbamoylphenyl OCH.sub.3 Br 14 4-carbamoylphenyl OCH.sub.3 I 15 3-carbamoylphenyl OCH.sub.3 Cl 16 3-carbamoylphenyl OCH.sub.3 Br 17 3-carbamoylphenyl OCH.sub.3 I

[Compound 1] 3-chloro-4-(3-chlorobenzyloxy)-5-methoxybenzaldehyde

[0108] 5-chlorovanillin (1.00 g, 5.36 mmol) and 3-chlorobenzyl chloride (0.70 g, 5.36 mmol) were used to obtain Compound 1 as an ivory solid (1.50 g, 96.5%).

[0109] R.sub.f 0.54 (ethyl acetate:n-hexane=1:3); .sup.1H-NMR (CDCl.sub.3, 400 MHz) δ 3.95 (s, 3H), 5.13 (s, 2H), 7.30-7.32 (m, 2H), 7.36 (d, J=2.0 Hz, 1H), 7.37-7.39 (m, 1H), 7.51 (d, J=2.0 Hz, 1H), 7.53-7.54 (m, 1H), 9.86 (s, 1H); .sup.13C-NMR (CDCl.sub.3, 100 MHz) 56.5, 74.3, 109.6, 125.9, 126.4, 128.5, 128.6, 129.4, 129.9, 132.9, 134.5, 138.7, 149.4, 154.5, 190.1 ppm.

[Compound 2] 3-bromo-4-(3-chlorobenzyloxy)-5-methoxybenzaldehyde

[0110] 5-bromovanillin (1.00 g, 4.33 mmol) and 3-chlorobenzyl chloride (0.58 g, 5.36 mmol) were used to obtain Compound 2 as an ivory solid (1.50 g, 96.5%).

[0111] R.sub.f 0.65 (ethyl acetate:n-hexane=1:3); .sup.1H-NMR (CDCl.sub.3, 400 MHz) δ 3.94 (s, 3H), 5.12 (s, 2H), 7.30-7.32 (m, 2H), 7.38 (dd, J=8.4, 1.2 Hz, 1H), 7.40 (d, J=2.0 Hz, 1H), 7.54-7.56 (m, 1H), 7.66 (d, J=2.0 Hz, 1H), 9.85 (s, 1H); .sup.13C-NMR (CDCl.sub.3, 100 MHz) 56.5, 74.2, 110.3, 118.5, 126.5, 128.6, 129.0, 129.9, 133.5, 134.5, 138.7, 150.4, 154.4, 190.0 ppm.

[Compound 3] 4-((3-chlorobenzyl)oxy)-3-iodo-5-methoxybenzaldehyde

[0112] 5-iodovanillin (1.00 g, 3.60 mmol) and 3-chlorobenzyl chloride (0.58 g, 5.36 mmol) were used to obtain Compound 3 as an ivory solid (1.423 g, 98.1%).

[0113] R.sub.f 0.67 (ethyl acetate:n-hexane=1:3); .sup.1H-NMR (CDCl.sub.3, 400 MHz) δ 3.94 (s, 3H), 5.10 (s, 2H), 7.31-7.33 (m, 2H), 7.41 (dd, J=8.4, 1.2 Hz, 1H), 7.43 (d, J=1.6 Hz, 1H), 7.57-7.58 (m, 1H), 7.87 (d, J=1.6 Hz, 1H), 9.84 (s, 1H); .sup.13C-NMR (CDCl.sub.3, 100 MHz) 56.4, 74.0, 92.8, 111.2, 126.6, 128.7, 128.8, 129.9, 134.4, 134.5, 135.1, 138.7, 152.9, 153.2, 189.9 ppm.

[Compound 4] ethyl 3-((2-chloro-4-formyl-6-methoxyphenoxy)methyl)benzoate

[0114] 5-chlorovanillin (1.00 g, 5.36 mmol) and ethyl 3-(chloromethyl)benzoate (1.06 g, 5.36 mmol) were used to obtain Compound 4 an a yellow solid (1.18 g, 63.1%).

[0115] R.sub.f 0.39 (ethyl acetate:n-hexane=1:3); .sup.1H-NMR (CDCl.sub.3, 400 MHz) δ 1.41 (t, J=7.2 Hz, 3H), 3.95 (s, 3H), 4.39 (q, J=7.2 Hz, 2H), 5.21 (s, 2H), 7.30 (ddd, J=7.6, 1.2, 1.2 Hz, 1H), 7.36 (d, J=1.6 Hz, 1H), 7.46 (dd, J=7.6, 7.6 Hz, 1H), 7.50 (d, J=2.0 Hz, 1H), 8.02 (ddd, J=7.6, 1.2, 1.2 Hz, 1H), 8.18 (dd, J=1.2, 1.2 Hz, 1H), 9.85 (s, 1H); .sup.13C-NMR (CDCl.sub.3, 100 MHz) 14.6, 56.5, 61.3, 74.7, 109.6, 126.0, 128.7, 129.4, 129.6, 129.7, 130.9, 132.8, 132.9, 137.1, 149.5, 154.6, 166.6, 190.2 ppm.

[Compound 5] ethyl 3-((2-bromo-4-formyl-6-methoxyphenoxy)methyl)benzoate

[0116] 5-bromovanillin (1.00 g, 4.33 mmol) and ethyl 3-(chloromethyl)benzoate (0.86 g, 4.33 mmol) were used to obtain Compound 5 as a pale yellow solid (1.53 g, 90.1%).

[0117] R.sub.f 0.35 (ethyl acetate:n-hexane=1:3); .sup.1H-NMR (CDCl.sub.3, 400 MHz) δ 1.41 (t, J=7.2 Hz, 3H), 3.95 (s, 3H), 4.39 (q, J=7.2 Hz, 2H), 5.21 (s, 2H), 7.40 (d, J=2.0 Hz, 1H), 7.46 (dd, J=7.6, 7.6 Hz, 1H), 7.66 (d, J=2.0 Hz, 1H), 7.75 (ddd, J=7.6, 1.2, 1.2 Hz, 1H), 8.02 (ddd, J=7.6, 1.2, 1.2 Hz, 1H), 8.19 (dd, J=1.2, 1.2 Hz, 1H), 9.85 (s, 1H); .sup.13C-NMR (CDCl.sub.3, 100 MHz) 14.6, 56.5, 61.3, 74.6, 110.3, 118.6, 128.7, 129.0, 129.7, 130.9, 133.0, 133.4, 137.1, 150.6, 154.4, 166.6, 190.0 ppm.

[Compound 6] ethyl 3-((4-formyl-2-iodo-6-methoxyphenoxy)methyl)benzoate

[0118] 5-iodovanillin (1.00 g, 3.60 mmol) and ethyl 3-(chloromethyl)benzoate (0.71 g, 3.60 mmol) were used to obtain Compound 6 as a pale yellow solid (1.41 g, 88.8%).

[0119] R.sub.f 0.39 (ethyl acetate:n-hexane=1:3); .sup.1H-NMR (CDCl.sub.3, 400 MHz) δ 1.41 (t, J=7.2 Hz, 3H), 3.95 (s, 3H), 4.39 (q, J=7.2 Hz, 2H), 5.21 (s, 2H), 7.43 (d, J=1.6 Hz, 1H), 7.47 (dd, J=7.6, 7.6 Hz, 1H), 7.78 (ddd, J=7.6, 1.2, 1.2 Hz, 1H), 7.86 (d, J=1.6 Hz, 1H), 8.02 (ddd, J=7.6, 1.2, 1.2 Hz, 1H), 8.22 (dd, J=1.6, 1.6 Hz, 1H), 9.83 (s, 1H); .sup.13C-NMR (CDCl.sub.3, 100 MHz) 14.6, 56.3, 61.3, 74.4, 92.8, 111.2, 128.7, 129.7, 129.8, 130.9, 133.1, 134.3, 135.1, 137.1, 152.9, 153.2, 166.6, 189.9 ppm.

[Compound 7] 3-((2-chloro-4-formyl-6-methoxyphenoxy)methyl)benzonitrile

[0120] 5-chlorovanillin (1.00 g, 5.36 mmol) and ethyl 3-(bromomethyl)benzonitrile (1.05 g, 5.36 mmol) were used to obtain Compound 7 as a white solid (1.30 g, 80.5%).

[0121] R.sub.f 0.25 ethyl acetate:n-hexane=1:3); .sup.1H-NMR (CDCl.sub.3, 400 MHz) δ 3.96 (s, 3H), 5.17 (s, 2H), 7.38 (d, J=2.0 Hz, 1H), 7.50 (dd, J=7.6, 7.6 Hz, 1H), 7.52 (d, J=2.0 Hz, 1H), 7.64 (ddd, J=7.6, 1.6, 1.6 Hz, 1H), 7.75 (ddd, J=7.6, 1.6, 1.6 Hz, 1H), 7.84 (dd, J=1.6, 1.2 Hz, 1H), 9.87 (s, 1H); .sup.13C-NMR (CDCl.sub.3, 100 MHz) 56.6, 73.8, 109.6, 112.8, 118.9, 126.0, 129.4, 129.5, 131.8, 132.1, 132.5, 133.1, 138.4, 149.1, 154.5, 190.1 ppm.

[Compound 8] 3-((2-bromo-4-formyl-6-methoxyphenoxy)methyl)benzonitrile

[0122] 5-bromovanillin (1.00 g, 4.33 mmol) and ethyl 3-(bromomethyl)benzonitrile (0.85 g, 4.33 mmol) were used to obtain Compound 8 as a white solid (1.53 g, 83.8%).

[0123] R.sub.f 0.24 (ethyl acetate:n-hexane=1:3); .sup.1H-NMR (CDCl.sub.3, 400 MHz) δ 3.95 (s, 3H), 5.16 (s, 2H), 7.42 (d, J=2.0 Hz, 1H), 7.50 (dd, J=8.0, 7.6 Hz, 1H), 7.64 (ddd, J=7.6, 2.0 1.6 Hz, 1H), 7.67 (d, J=1.6 Hz, 1H), 7.70 (ddd, J=8.0, 1.6, 1.2 Hz, 1H), 7.86 (dd, J=1.6, 1.6 Hz, 1H), 9.86 (s, 1H); .sup.13C-NMR (CDCl.sub.3, 100 MHz) 56.5, 73.6, 110.3, 112.8, 118.5, 118.9, 129.0, 129.5, 131.9, 132.1, 132.6, 133.7, 138.4, 150.1, 154.3, 189.9 ppm.

[Compound 9] 3-((4-formyl-2-iodo-6-methoxyphenoxy)methyl)benzonitrile

[0124] 5-iodovanillin (1.00 g, 3.60 mmol) and ethyl 3-(bromomethyl)benzonitrile (0.71 g, 3.60 mmol) were used to obtain Compound 9 as a white solid (1.20 g, 84.4%).

[0125] R.sub.f 0.27 (ethyl acetate:n-hexane=1:3); .sup.1H-NMR (CDCl.sub.3, 400 MHz) δ 3.95 (s, 3H), 5.15 (s, 2H), 7.44 (d, J=2.0 Hz, 1H), 7.51 (dd, J=8.0, 7.6 Hz, 1H), 7.65 (ddd, J=7.6, 1.6, 1.2 Hz, 1H), 7.78 (ddd, J=7.6, 1.6, 1.2 Hz, 1H), 7.88 (d, J=2.0 Hz, 1H), 7.89 (dd, J=1.6, 1.2 Hz, 1H), 9.85 (s, 1H); .sup.13C-NMR (CDCl.sub.3, 100 MHz) 56.4, 73.4, 92.7, 111.3, 112.8, 118.9, 129.5, 132.0, 132.1, 132.7, 134.6, 135.0, 138.3, 152.5, 153.2, 189.8 ppm.

[Compound 10] 3-chloro-4-((3-fluorobenzyl)oxy)-5-methoxybenzaldehyde

[0126] 5-chlorovanillin (1.00 g, 5.36 mmol) and ethyl 3-fluorobenzyl chloride (0.78 g, 5.36 mmol) were used to obtain Compound 10 as a white solid (1.15 g, 72.8%).

[0127] R.sub.f 0.58 (ethyl acetate:n-hexane=1:3); .sup.1H-NMR (CDCl.sub.3, 400 MHz) δ 3.95 (s, 3H), 5.15 (s, 2H), 7.00-7.05 (m, 1H), 7.24-7.28 (m, 2H), 7.34 (dd, J=8.0, 2.0 Hz, 1H), 7.37 (d, J=2.0 Hz, 1H), 7.51 (d, J=2.0 Hz, 1H), 9.85 (s, 1H); .sup.13C-NMR (CDCl.sub.3, 100 MHz) 56.5, 74.4, 109.7, 115.3, 115.5, 123.8, 126.0, 130.1, 132.9, 139.3, 149.5, 154.6, 161.8, 164.3, 190.2 ppm.

[Compound 11] 4-((3-chlorobenzyl)oxy)-3-methoxybenzaldehyde

[0128] Vanillin (1.00 g, 6.57 mmol) and 3-methoxybenzyl chloride (1.03 g, 6.57 mmol) were used to obtain Compound 11 as a yellow semisolid (1.48 g, 99.6%).

[0129] R.sub.f 0.45 (ethyl acetate:n-hexane=1:3); .sup.1H-NMR (CDCl.sub.3, 400 MHz) δ 3.95 (s, 3H), 5.19 (s, 2H), 6.95 (d, J=8.4 Hz, 1H), 7.29-7.32 (m, 3H), 7.39 (dd, J=8.0, 1.6 Hz, 1H), 7.43-7.44 (m, 2H), 9.84 (s, 1H); .sup.13C-NMR (CDCl.sub.3, 100 MHz) 56.2, 70.2, 109.7, 112.6, 125.4, 126.6, 127.4, 128.6, 130.2, 130.7, 134.8, 138.3, 150.3, 153.4, 191.0 ppm.

[Compound 12] 4-((2-chloro-4-formyl-6-methoxyphenoxy)methyl)benzamide

[0130] 5-chlorovanillin (0.18 g, 0.96 mmol) and 4-(chloromethyl)benzamide (0.16 g, 0.96 mmol) were used to obtain Compound 12 as a white solid (0.22 g, 73.2%).

[0131] R.sub.f 0.25 (ethyl acetate:n-hexane=1:1); .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ 3.88 (s, 3H), 5.12 (s, 2H), 7.29 (d, J=1.6 Hz, 1H), 7.44 (d, J=2.0 Hz, 1H), 7.48 (d, J=8.0 Hz, 2H), 7.83 (d, J=8.0 Hz, 2H), 9.78 (s, 1H); .sup.13C-NMR (400 MHz, DMSO-d.sub.6) 55.2, 73.7, 109.5, 124.5, 127.2, 127.4, 128.3, 132.0, 133.2, 139.4, 148.4, 153.7, 168.4, 189.4 ppm.

[Compound 13] 4-((2-bromo-4-formyl-6-methoxyphenoxy)methyl)benzamide

[0132] 5-bromovanillin (0.18 g, 0.79 mmol) and 4-(chloromethyl)benzamide (0.13 g, 0.79 mmol) were used to obtain Compound 13 as a white solid (0.20 g, 73.2%).

[0133] R.sub.f 0.16 (ethyl acetate:n-hexane=1:1); .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ 3.88 (s, 3H), 5.11 (s, 2H), 7.34 (d, J=2.0 Hz, 1H), 7.50 (d, J=8.4 Hz, 2H), 7.60 (d, J=1.6 Hz, 1H), 7.83 (d, J=8.4 Hz, 2H), 9.78 (s, 1H); .sup.13C-NMR (400 MHz, DMSO-d.sub.6) 55.7, 73.6, 100.1, 117.6, 127.2, 127.4, 127.5, 132.6, 133.2, 139.4, 149.5, 153.5, 168.4, 189.3 ppm.

[Compound 14] 4-((4-formyl-2-iodo-6-methoxyphenoxy)methyl)benzamide

[0134] 5-iodovanillin (0.26 g, 0.94 mmol) and 4-(chloromethyl)benzamide (0.16 g, 0.94 mmol) were used to obtain Compound 14 as a white solid (0.28 g, 72.4%).

[0135] R.sub.f 0.10 (ethyl acetate:n-hexane=1:1); .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ 3.87 (s, 3H), 5.10 (s, 2H), 7.36 (d, J=2.0 Hz, 1H), 7.53 (d, J=8.0 Hz, 2H), 7.79 (d, J=1.6 Hz, 1H), 7.83 (d, J=8.4 Hz, 2H), 9.76 (s, 1H); .sup.13C-NMR (400 MHz, DMSO-d.sub.6) 55.6, 73.4, 92.1, 111.0, 127.2, 127.6, 133.1, 133.5, 133.6, 139.4, 151.9, 152.3, 168.4, 189.1 ppm.

[Compound 15] 3-((2-chloro-4-formyl-6-methoxyphenoxy)methyl)benzamide

[0136] 5-chlorovanillin (0.40 g, 2.14 mmol) and 3-(chloromethyl)benzamide (0.36 g, 2.14 mmol) were used to obtain Compound 15 as a white solid (0.32 g, 45.0%).

[0137] R.sub.f 0.10 (ethyl acetate:n-hexane=1:1); .sup.1H-NMR (CDCl.sub.3, 400 MHz) δ 3.88 (s, 3H), 5.11 (s, 2H), 7.30 (d, J=2.0 Hz, 1H), 7.37 (dd, J=7.6, 7.6 Hz, 1H), 7.44 (d, J=2.0 Hz, 1H), 7.59 (d, J=8.0 Hz, 1H), 7.77 (ddd, J=7.6, 1.6, 1.2 Hz, 1H), 794 (dd, J=1.6, 1.6 Hz, 1H), 9.78 (s, 1H); .sup.13C-NMR (400 MHz, DMSO-d.sub.6) 56.4, 74.1, 110.9, 124.1, 127.2, 127.6, 127.9, 128.2, 131.1, 132.6, 134.4, 136.6, 148.2, 154.0, 167.6, 191.1 ppm.

[Compound 16] 3-((2-bromo-4-formyl-6-methoxyphenoxy)methyl)benzamide

[0138] 5-bromovanillin (0.18 g, 0.79 mmol) and 3-(chloromethyl)benzamide (0.13 g, 0.79 mmol) were used to obtain Compound 16 as a white solid (0.20 g, 73.2%).

[0139] R.sub.f 0.16 (ethyl acetate:n-hexane=1:1); .sup.1H-NMR (DMSO-d.sub.6, 400 MHz) δ 3.88 (s, 3H), 5.10 (s, 2H), 7.34 (d, J=2.0 Hz, 1H), 7.38 (dd, J=8.0, 8.0 Hz, 1H), 7.60 (d, J=2.0 Hz, 1H), 7.62 (ddd, J=7.6, 1.2, 1.2 Hz 1H), 7.78 (ddd, J=7.6, 1.2, 1.2 Hz 1H), 7.96 (dd, J=1.2, 1.2 Hz, 1H), 9.78 (s, 1H); .sup.13C-NMR (400 MHz, DMSO-d.sub.6) 55.7, 73.8, 110.0, 117.7, 126.9, 127.2, 127.6, 127.9, 131.0, 132.6, 133.5, 136.3, 149.5, 153.6, 168.5, 189.3 ppm.

[Compound 17] 3-((4-formyl-2-iodo-6-methoxyphenoxy)methyl)benzamide

[0140] 5-iodovanillin (0.26 g, 0.94 mmol) and 3-(chloromethyl)benzamide (0.16 g, 0.94 mmol) were used to obtain Compound 17 as a white solid (0.28 g, 72.40%).

[0141] R.sub.f 0.10 (ethyl acetate:n-hexane=1:1); .sup.1H-NMR (DMSO-d.sub.6, 400 MHz) δ 3.88 (s, 3H), 5.09 (s, 2H), 7.36 (d, J=2.0 Hz, 1H), 7.38 (dd, J=8.0, 8.0 Hz, 1H), 7.65 (ddd, J=7.6, 1.6, 1.6 Hz 1H), 7.78 (ddd, J=7.6, 1.2, 1.2 Hz, 1H), 7.79 (d, J=2.0 Hz, 1H), 7.98 (dd, J=1.6, 1.6 Hz, 1H), 9.76 (s, 1H); .sup.13C-NMR (400 MHz, DMSO-d.sub.6) 55.6, 73.6, 92.2, 111.0, 126.9, 127.3, 127.8, 131.1, 133.5, 113.6, 133.7, 136.2, 151.9, 152.4, 168.5, 189.1 ppm.

[Synthesis Method 2] Synthesis Examples 1 to 19 (A1 to A19)

[0142] The O-benzylated compound (1 equivalent) synthesized in Synthesis Method 1; and 1 equivalent of 1-(2-aminoethyl)piperidine; 2-(2-aminoethyl)pyridine; aminoethyl)thiophene; or (4-carboxyphenyl)methylamine were refluxed for 2 hours using ethanol as a solvent, and then cooled to room temperature. Thereafter, the solvent was completely removed under reduced pressure, and Synthesis Examples 1 to 19 corresponding to the imine compounds were obtained (see Table 3)

TABLE-US-00003 TABLE 3 Synthesis Example R R.sub.1 R.sub.2 R.sub.3 1 3-chlorophenyl OCH.sub.3 Br N-piperidinyl 2 3-chlorophenyl OCH.sub.3 I N-piperidinyl 3 3-ethoxycarbonylphenyl OCH.sub.3 Cl N-piperidinyl 4 3-ethoxycarbonylphenyl OCH.sub.3 Br N-piperidinyl 5 3-ethoxycarbonylphenyl OCH.sub.3 I N-piperidinyl 6 3-cyanophenyl OCH.sub.3 Cl N-piperidinyl 7 3-cyanophenyl OCH.sub.3 Br N-piperidinyl 8 3-cyanophenyl OCH.sub.3 I N-piperidinyl 9 3-fluorophenyl OCH.sub.3 Cl N-piperidinyl 10 3-chlorophenyl OCH.sub.3 Cl 2-thiophenyl 11 3-chlorophenyl OCH.sub.3 Cl 2-pyridyl 12 3-chlorophenyl OCH.sub.3 H 2-thiophenyl 13 3-chlorophenyl OCH.sub.3 H 2-pyridyl 14 4-carbamoylphenyl OCH.sub.3 Cl N-piperidinyl 15 4-carbamoylphenyl OCH.sub.3 Br N-piperidinyl 16 4-carbamoylphenyl OCH.sub.3 I N-piperidinyl 17 3-carbamoylphenyl OCH.sub.3 Cl N-piperidinyl 18 3-carbamoylphenyl OCH.sub.3 Cl 2-thiophenyl 19 3-chlorophenyl OCH.sub.3 Cl 4-carboxyphenyl

[Synthesis Example 1] (E)-1-(3-bromo-4-((3-chlorobenzyl)oxy)-5-methoxyphenyl)-N-(2-(pyridine-1-1.48yl)ethyl)methanimine

[0143] Compound 2 (208 mg, 0.59 mmol) and 1-(2-aminoethyl)piperidine (75 mg, 0.59 mmol) were used to obtain Synthesis Example 1 as a brown syrup (260 mg, 95.4%).

[0144] .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 1.48-1.51 (m, 2H), 1.67-1.72 (m, 4H), 2.61-2.63 (m, 4H), 2.77 (t, J=7.2 Hz, 2H), 3.85 (t, J=6.8 Hz, 2H), 3.92 (s, 3H), 5.03 (s, 2H), 7.29-7.32 (m, 2H), 7.33 (d, J=1.6 Hz, 1H), 7.39 (ddd, J=8.8, 1.6, 1.6 Hz, 1H), 7.42 (d, J=1.6 Hz, 1H), 7.54-7.56 (m, 1H), 8.19 (dd, J=1.6, 1.2 Hz, 1H); .sup.13C-NMR (100 MHz, CDCl.sub.3) 23.9, 25.3, 54.4, 54.9, 56.4, 59.3, 74.1, 110.1, 118.2, 125.9, 126.5, 128.5, 128.7, 129.8, 133.5, 134.4, 139.1, 147.3, 154.1, 160.8 ppm.

[Synthesis Example 2] (E)-1-(4-((3-chlorobenzyl)oxy)-3-iodo-5-methoxyphenyl)-N-(2-(piperidine-1-yl)ethyl)methanimine

[0145] Compound 3 (205 mg, 0.51 mmol) and 1-(2-aminoethyl)piperidine (55 mg, 0.51 mmol) were used to obtain Synthesis Example 2 as a brown syrup (260 mg, 99.6%).

[0146] .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 1.48-1.51 (m, 2H), 1.65-1.70 (m, 4H), 2.57-2.61 (m, 4H), 2.75 (t, J=7.2 Hz, 2H), 3.83 (t, J=6.8 Hz, 2H), 3.91 (s, 3H), 5.01 (s, 2H), 7.30-7.32 (m, 2H), 7.37 (d, J=1.6 Hz, 1H), 7/41-7.43 (m, 1H), 7.57-7.59 (m, 1H), 7.62 (d, J=1.6 Hz, 1H), 8.17 (dd, J=1.6, 1.6 Hz, 1H); .sup.13C-NMR (100 MHz, CDCl.sub.3) 23.9, 25.3, 54.4, 54.9, 56.3, 59.3, 73.9, 92.8, 111.2, 111.3, 126.6, 128.5, 128.8, 129.8, 131.7, 134.4, 139.1, 149.8, 153.0, 160.5 ppm.

[Synthesis Example 3] Ethyl (E)-3-((2-chloro-6-methoxy-4-(((2-(piperidine-1-yl)ethyl)imido)methyl)phenoxy)methyl)benzoate

[0147] Compound 4 (200 mg, 0.57 mmol) and 1-(2-aminoethyl)piperidine (73.5 mg, 0.57 mmol) were used to obtain Synthesis Example 3 as a yellow syrup (275 mg, quantitative).

[0148] R.sub.f 0.18 (ethyl acetate:n-hexane=1:3); .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 1.40 (t, J=7.2 Hz, 3H), 1.44-1.48 (m, 2H), 1.59-1.63 (m, 4H), 2.50-2.53 (m, 4H), 2.67 (t, J=7.2 Hz, 2H), 3.78 (t, J=6.8 Hz, 2H), 3.92 (s, 3H), 4.38 (q, J=7.2 Hz, 2H), 5.12 (s, 2H), 7.24 (d, J=2.0 Hz, 1H), 7.29 (d, J=2.0 Hz, 1H), 7.40 (ddd, J=8.0, 2.0, 2.0 Hz, 1H), 7.44 (dd, J=8.0, 8.0 Hz, 1H), 8.00 (ddd, J=8.0, 2.0, 2.0 Hz, 1H), 8.16 (br s, 1H), 8.18 (dd, J=2.0, 2.0 Hz, 1H).

[Synthesis Example 4] Ethyl (E)-3-((2-bromo-6-methoxy-4-(((2-(piperidine-1-yl)ethyl)imino)methyl)phenoxy) methyl)benzoate

[0149] Compound 5 (200 mg, 0.51 mmol) and 1-(2-aminoethyl)piperidine (65.2 mg, 0.51 mmol) were used to obtain Synthesis Example 4 as a yellow syrup (252 mg, 98.2%).

[0150] R.sub.f 0.18 (ethyl acetate:n-hexane=1:3); .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 1.39 (t, J=7.2 Hz, 3H), 1.44-1.48 (m, 2H), 1.59-1.65 (m, 4H), 2.52-2.54 (m, 4H), 2.68 (t, J=7.2 Hz, 2H), 3.76 (t, J=6.8 Hz, 2H), 3.92 (s, 3H), 4.38 (q, J=7.2 Hz, 2H), 5.11 (s, 2H), 7.34 (d, J=1.6 Hz, 1H), 7.40 (d, J=1.6 Hz, 1H), 7.45 (dd, J=8.0, 8.0 Hz, 1H), 7.76 (ddd, J=8.0, 1.2, 1.2 Hz, 1H), 8.01 (ddd, J=8.0, 1.2, 1.2 Hz, 1H), 8.16 (br S, 1H), 8.20 (dd, J=1.2, 1.2 Hz, 1H).

[Synthesis Example 5] Ethyl (E)-3-((2-iodo-6-methoxy-4-(((2-(piperidine-1-yl)ethyl)imino)methyl)phenoxy) methyl)benzoate

[0151] Compound 6 (200 mg, 0.45 mmol) and 1-(2-aminoethyl)piperidine (58.3 mg, 0.45 mmol) were used to obtain Synthesis Example 5 as a yellow syrup (238 mg, 93.0%).

[0152] R.sub.f 0.21 (ethyl acetate:n-hexane=1:3); .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 1.40 (t, J=7.2 Hz, 3H), 1.43-1.46 (m, 2H), 1.59-1.64 (m, 4H), 2.47-2.51 (m, 4H), 2.67 (t, J=7.2 Hz, 2H), 3.77 (t, J=6.8 Hz, 2H), 3.91 (s, 3H), 4.38 (q, J=7.2 Hz, 2H), 5.10 (s, 2H), 7.37 (d, J=2.0 Hz, 1H), 7.45 (dd, J=8.0, 8.0 Hz, 1H), 7.61 (d, J=2.0 Hz, 1H), 7.7 (ddd, J=7.6, 1.6, 1.6 Hz, 1H), 8.01 (ddd, J=7.6, 1.6, 1.6 Hz, 1H), 8.15 (br s, 1H), 8.22 (dd, J=1.2, 1.2 Hz, 1H); .sup.13C-NMR (100 MHz, CDCl.sub.3) 14.6, 24.4, 26.0, 55.1, 56.3, 59.1, 59.8, 61.2, 74.2, 92.7, 110.2, 129.6, 129.5, 129.8, 130.8, 131.8, 133.1, 134.6, 137.4, 149.8, 153.0, 160.0, 166.7 ppm.

[Synthesis Example 6] (E)-3-((2-chloro-6-methoxy-4-(((2-(piperidine-1-yl)ethyl)imino)methyl)phenoxy)methyl) benzonitrile

[0153] Compound 7 (200 mg, 0.66 mmol) and 1-(2-aminoethyl)piperidine (85.0 mg, 0.66 mmol) were used to obtain Synthesis Example 6 as a yellow syrup (273 mg, quantitative).

[0154] R.sub.f 0.15 (ethyl acetate:n-hexane=1:3); .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 1.40-1.46 (m, 2H), 1.54-1.62 (m, 4H), 2.47-2.49 (m, 4H), 2.64 (t, J=7.6 Hz, 2H), 3.76 (t, J=7.6 Hz, 2H), 3.91 (s, 3H), 5.07 (s, 2H), 7.24 (d, J=1.6 Hz, 1H), 7.30 (d, J=1.6 Hz, 1H), 7.44 (d, J=7.6 Hz, 1H), 7.47 (dd, J=8.0, 7.6 Hz, 1H), 7.60 (ddd, J=7.6, 1.2, 1.2 Hz, 1H), 7.83 (br s, 1H), 8.18 (br s, 1H).

[Synthesis Example 7] (E)-3-((2-bromo-6-methoxy-4-(((2-(piperidine-1-yl)ethyl)imino)methyl)phenoxy)methyl)benzonitrile

[0155] Compound 8 (200 mg, 0.58 mmol) and 1-(2-aminoethyl)piperidine (74.1 mg, 0.58 mmol) were used to obtain Synthesis Example 7 as a yellow syrup (265 mg, quantitative).

[0156] R.sub.f 0.15 (ethyl acetate:n-hexane=1:3); .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 1.42-1.46 (m, 2H), 1.55-1.62 (m, 4H), 2.46-2.49 (m, 4H), 2.64 (t, J=7.6 Hz, 2H), 3.74 (t, J=7.6 Hz, 2H), 3.91 (s, 3H), 5.06 (s, 2H), 7.34 (d, J=1.6 Hz, 1H), 7.40 (d, J=1.6 Hz, 1H), 7.47 (dd, J=8.0, 7.6 Hz, 1H), 7.61 (ddd, J=7.6, 1.6, 1.2 Hz, 1H), 7.75 (ddd, J=7.6, 2.0, 1.2 Hz, 1H), 7.85 (dd, J=1.6, 1.2 Hz, 1H), 8.16 (br s, 1H).

[Synthesis Example 8] (E)-3-((2-iodo-6-methoxy-4-(((2-(piperidine-1-yl)ethyl)imino)methyl)phenoxy)methyl)benzonitrile

[0157] Compound 9 (200 mg, 0.51 mmol) and 1-(2-aminoethyl)piperidine (65.2 mg, 0.51 mmol) were used to obtain Synthesis Example 8 as a yellow syrup (245 mg, 95.7%).

[0158] R.sub.f 0.14 (ethyl acetate:n-hexane=1:3); .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 1.42-1.48 (m, 2H), 1.58-1.62 (m, 4H), 2.48-2.52 (m, 4H), 2.66 (t, J=7.2 Hz, 2H), 3.77 (t, J=7.2 Hz, 2H), 3.91 (s, 3H), 5.06 (s, 2H), 7.38 (d, J=1.6 Hz, 1H), 7.49 (dd, J=8.0, 7.6 Hz, 1H), 7.61 (d, J=1.6 Hz, 1H), 7.62 (d, J=8.0 Hz, 1H), 7.79 (d, J=8.0 Hz, 1H), 7.88 (br s, 1H), 8.16 (br s, 1H); .sup.13C-NMR (100 MHz, CDCl.sub.3) 24.4, 26.1, 55.1, 56.3, 59.1, 59.8, 73.3, 92.6, 110.9, 112.7, 119.0, 129.4, 131.8, 131.9, 132.0, 132.7, 134.9, 138.7, 149.3, 152.9, 159.9 ppm.

[Synthesis Example 9] (E)-1-(3-chloro-4-((3-fluorobenzyl)oxy-5-methoxyphenyl)-N-(2-(piperidine-1-yl)ethyl)methanimine

[0159] Compound 10 (200 mg, 0.68 mmol) and 1-(2-aminoethyl)piperidine (87.0 mg, 0.68 mmol) were used to obtain Synthesis Example 9 as a yellow syrup (267 mg, 97.2%).

[0160] .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 1.44-1.49 (m, 2H), 1.60-1.66 (m, 4H), 2.51-2.54 (m, 4H), 2.69 (t, J=7.2 Hz, 2H), 3.80 (t, J=7.2 Hz, 2H), 3.92 (s, 3H), 5.06 (s, 2H), 7.02 (dd, J=7.6, 7.2 Hz, 1H), 7.25-7.33 (m, 5H), 8.18 (br s, 1H); .sup.13C-NMR (100 MHz, CDCl.sub.3) 24.2, 25.8, 55.0, 56.4, 58.8, 59.6, 74.2, 109.2, 115.3, 115.5, 123.1, 128.9, 130.8, 133.1, 139.7, 146.2, 154.6, 160.6, 161.8, 164.3 ppm.

[Synthesis Example 10] (E)-1-(3-chloro-4-((3-chlorobenzyl)oxy)-5-methoxyphenyl)-N-(2-(thiophene-2-yl)ethyl)methanimine

[0161] Compound 1 (200 mg, 0.64 mmol) and 2-(2-aminoethyl)thiophene (87.6 mg, 0.64 mmol) were used to obtain Synthesis Example 10 as a brown syrup (270 mg, quantitative).

[0162] R.sub.f 0.68 (ethyl acetate:n-hexane=1:3); .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 3.24 (t, J=7.2 Hz, 2H), 3.88 (t, J=7.2 Hz, 2H), 3.93 (s, 3H), 5.05 (s, 2H), 6.83-6.84 (m, 1H), 6.92 (dd, J=6.0, 3.6 Hz, 1H), 7.14 (dd, J=3.6, 1.6 Hz, 1H), 7.23 (d, J=2.0 Hz, 1H), 7.29-7.32 (m, 2H), 7.34 (br s, 1H), 7.36-7.39 (m, 1H), 7.54-7.55 (m, 1H), 8.07 (s, 1H); .sup.13C-NMR (100 MHz, CDCl.sub.3) 31.6, 56.5, 62.9, 74.2, 109.6, 123.9, 126.0, 126.4, 126.5, 127.0, 128.5, 128.6, 128.7, 128.9, 129.8, 132.9, 134.4, 138.8, 154.3, 154.6, 160.3 ppm.

[Synthesis Example 11] (E)-1-(3-chloro-4-((3-chlorobenzyl)oxy)-5-methoxybenzyl)-N-(2-(pyridine-2-yl)ethyl)methanimine

[0163] Compound 1 (100 mg, 0.32 mmol) and 12-(2-aminoethyl)pyridine (40 mg, 0.32 mmol) were used to obtain Synthesis Example 11 as a brown syrup (133 mg, quantitative).

[0164] .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ 3.07 (t, J=7.6 Hz, 2H), 3.88 (s, 3H), 3.93 (t, J=7.6 Hz, 2H), 5.06 (s, 2H), 7.20 (ddd, J=7.2, 4.8, 1.2 Hz, 1H), 7.28 (d, J=7.6 Hz, 1H), 7.35 (d, J=2.0 Hz, 1H), 7.40 (d, J=2.0 Hz, 1H), 7.41-7.43 (m, 3H), 7.54 (dd, J=1.2, 1.2 Hz, 1H), 7.69 (ddd, J=7.6, 7.6, 1.2 Hz, 1H), 8.24 (s, 1H), 8.50 (ddd, J=4.8, 1.6, 0.8 Hz, 1H); .sup.13C-NMR (100 MHz, DMSO-d.sub.6) 56.2, 59.9, 73.3, 121.3, 121.4, 123.3, 126.7, 127.3, 127.8, 128.0, 130.2, 132.98, 132.9, 136.3, 139.3, 144.9, 149.0, 153.7, 159.3, 159.5, ppm.

[Synthesis Example 12] (E)-1-(4-((3-Chlorobenzyl)oxy)-3-methoxyphenyl)-N-(2-(thiophen-2-yl)ethyl) methanimine

[0165] Compound 11 (200 mg, 0.72 mmol) and 2-2(aminoethyl)thiophene (91.9 mg, 0.72 mmol) were used to obtain Synthesis Example 12 as a brown syrup (280 mg, quantitative).

[0166] .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 3.24 (t, J=7.2 Hz, 2H), 3.86 (t, J=7.2 Hz, 2H), 3.97 (s, 3H), 5.16 (s, 2H), 6.84 (dd, J=2.4, 1.2 Hz, 1H), 6.86 (d, J=8.0 Hz, 1H), 6.91 (dd, J=5.2, 3.2 Hz, 1H), 7.08 (dd, J=8.4, 2.0 Hz, 1H), 7.13 (dd, J=5.2, 1.2 Hz, 1H), 7.28-7.32 (m, 3H), 7.44 (dd, J=1.6, 1.6 Hz, 1H), 7.52 (br s, 1H), 8.08 (s, 1H); .sup.13C-NMR (100 MHz, CDCl.sub.3) 31.8, 56.3, 62.9, 70.3, 109.6, 113.2, 123.1, 123.8, 125.3, 125.4, 126.9, 127.5, 128.4, 130.1, 130.3, 134.8, 139.0, 142.6, 150.2, 150.4, 161.5 ppm.

[Synthesis Example 13] (E)-1-(4-((3-chlorobenzyl)oxy)-3-methoxyphenyl)-N-(2-(pyridine-2-yl)ethyl)methanimine

[0167] Compound 11 (200 mg, 0.72 mmol) and 2-(2-aminoethyl)thiophene (88.3 mg, 0.72 mmol) were used to obtain Synthesis Example 13 as a brown syrup (277 mg, quantitative).

[0168] .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 3.18 (t, J=7.2 Hz, 2H), 3.94 (s, 3H), 3.99 (t, J=7.2 Hz, 2H), 5.14 (s, 2H), 6.83 (dd, J=8.4 Hz, 1H), 7.03 (dd, J=8.4, 2.0 Hz, 1H), 7.10 (ddd, J=8.8, 4.8, 1.2 Hz, 1H), 7.18 (d, J=7.6 Hz, 1H), 7.27-7.321 (m, 3H), 7.41-7.44 (m, 2H), 7.57 (ddd, J=8.0, 7.6, 2.0 Hz, 1H), 8.11 (br s, 1H), 8.55 (ddd, J=4.8, 1.2, 1.2 Hz, 1H); .sup.13C-NMR (100 MHz, CDCl.sub.3) 40.0, 56.2, 61.2, 70.3, 109.5, 113.2, 121.5, 123.0, 123.9, 125.4, 127.5, 128.4, 130.1, 130.2, 134.8, 136.4, 139.0, 149.6, 150.1, 150.3, 160.1, 161.4 ppm.

[Synthesis Example 14] (E)-4-((2-chloro-6-methoxy-4-(((2-(piperidine-1-yl)ethyl)imido)methyl)phenoxy)methyl)benzamide

[0169] Compound 12 (100 mg, 0.31 mmol) and 1-(2-aminoethyl)piperidine (40.1 mg, 0.31 mmol) were used to obtain Synthesis Example 14 as a pink solid (132 mg, 98.6%).

[0170] R.sub.f 0.10 (ethyl acetate:n-hexane=1:1); .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 1.43-1.49 (m, 2H), 1.60-1.67 (m, 4H), 2.52-2.54 (m, 4H), 2.69 (t, J=7.2 Hz, 2H), 3.77 (t, J=6.8 Hz, 2H), 3.91 (s, 3H), 5.13 (s, 2H), 7.24 (d, J=2.4 Hz, 1H), 7.30 (d, J=2.4 Hz, 1H), 7.59 (d, J=8.0 Hz, 2H), 7.82 (d, J=8.0 Hz, 2H), 8.17 (br s, 1H); .sup.13C-NMR (100 MHz, CDCl.sub.3) 22.3, 22.7, 29.9, 54.0, 56.5, 74.5, 109.7, 126.0, 128.4, 129.4, 132.9, 133.3, 140.9, 149.4, 154.6, 168.9 ppm.

[Synthesis Example 15] (E)-4-((2-bromo-6-methoxy-4-(((2-(piperidine-1-yl)ethyl)imino)methyl)phenoxy)methyl)benzamide

[0171] Compound 13 (100 mg, 0.27 mmol) and 1-(2-aminoethyl)piperidine (35.2 mg, 0.27 mmol) were used to obtain Synthesis Example 15 as a pale yellow solid (123 mg, 97.3%).

[0172] R.sub.f 0.06 (ethyl acetate:n-hexane=1:1); .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 1.47-1.52 (m, 2H), 1.63-1.69 (m, 4H), 2.51-2.54 (m, 4H), 2.72 (t, J=7.2 Hz, 2H), 3.82 (t, J=6.8 Hz, 2H), 3.91 (s, 3H), 5.13 (s, 2H), 7.34 (d, J=1.6 Hz, 1H), 7.41 (d, J=1.6 Hz, 1H), 7.61 (d, J=8.4 Hz, 2H), 7.82 (d, J=8.4 Hz, 2H), 8.18 (br s, 1H).

[Synthesis Example 16] (E)-4-((2-iodo-6-methoxy-4-(((2-(piperidine-1-yl)ethyl)imino)methyl)phenoxy)methyl)benzamide

[0173] Compound 14 (100 mg, 0.24 mmol) and 1-(2-aminoethyl)piperidine (31.2 mg, 0.24 mmol) were used to obtain Synthesis Example 16 as a pale yellow solid (128 mg, quantitative).

[0174] R.sub.f 0.08 (ethyl acetate:n-hexane=1:1); .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 1.47-1.53 (m, 2H), 1.64-1.67 (m, 4H), 2.56-2.60 (m, 4H), 2.74 (t, J=6.8 Hz, 2H), 3.83 (t, J=7.2 Hz, 2H), 3.91 (s, 3H), 5.11 (s, 2H), 7.37 (d, J=1.6 Hz, 1H), 7.62 (d, J=1.6 Hz, 1H), 7.64 (d, J=8.0 Hz, 2H), 7.86 (d, J=8.4 Hz, 2H), 8.17 (br s, 1H).

[Synthesis Example 17] (E)-3-((2-chloro-6-methoxy-4-(((2-(piperidine-1-yl)ethyl)imino)methyl)phenoxy)methyl)benzamide

[0175] Compound 15 (100 mg, 0.31 mmol) and 1-(2-aminoethyl)piperidine (40 mg, 0.31 mmol) were used to obtain Synthesis Example 17 as a brown syrup (97 mg, 72.1%).

[0176] .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 1.41-1.44 (m, 2H), 1.58-1.63 (m, 4H), 2.58-2.62 (m, 4H), 2.81 (t, J=6.8 Hz, 2H), 3.86 (t, J=6.8 Hz, 2H), 3.92 (s, 3H), 5.13 (s, 2H), 7.26 (d, J=1.6 Hz, 1H), 7.29 (d, J=1.6 Hz, 1H), 7.46 (dd, J=7.6, 7.6 Hz, 1H), 7.69 (d, J=7.6 Hz, 1H), 7.79 (dd, J=7.6, 1.6 Hz, 1H), 7.96 (br s, 1H), 8.19 (br s, 1H); .sup.13C-NMR (100 MHz, CDCl.sub.3) 22.8, 23.4, 54.2, 54.8, 56.5, 58.9, 74.5, 109.7, 122.9, 126.0, 127.3, 127.4, 128.9, 129.0, 132.1, 132.9, 133.7, 149.4, 154.3, 169.2 ppm.

[Synthesis Example 18] (E)-3-((2-Chloro-6-methoxy-4-(((2-(thiophen-2-yl)ethyl)imino)methyl)phenoxy)methyl)benzamide

[0177] Compound 15 (100 mg, 0.31 mmol) and 2-(2-aminoethyl)thiophene (39.8 mg, 0.31 mmol) were used to obtain Synthesis Example 18 as a foamy semisolid (93 mg, 69.3%).

[0178] .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 3.24 (t, J=6.8 Hz, 2H), 3.88 (t, J=6.8 Hz, 2H), 3.94 (s, 3H), 5.14 (s, 2H), 6.83 (dd, J=3.2, 1.2 Hz, 1H), 6.93 (dd, J=5.2, 3.6 Hz, 1H), 7.14 (dd, J=4.8, 1.2 Hz, 1H), 7.23 (d, J=1.6 Hz, 1H), 7.26 (d, J=1.6 Hz, 1H), 7.46 (dd, J=8.0, 8.0 Hz, 1H), 7.69 (d, J=7.6 Hz, 1H), 7.79 (dd, J=8.0, 1.6 Hz, 1H), 7.97 (br s, 1H), 8.06 (s, 1H); .sup.13C-NMR (100 MHz, CDCl.sub.3) 30.8, 56.6, 60.6, 74.6, 109.7, 124.1, 125.6, 126.0, 127.3, 127.4, 127.5, 128.9, 129.4, 131.9, 132.0, 132.9, 133.7, 137.5, 149.4, 154.6, 169.1 ppm.

[Synthesis Example 19] (E)-4-(((3-chloro-4-((3-chlorobenzyl)oxy)-5-methoxybenzylidene)amino)methyl)benzoic acid

[0179] Compound 1 (200 mg, 0.64 mmol), 4-(aminomethyl)benzoic acid (39.8 mg, 0.31 mmol), and triethylamine (65 mg, 0.64 mmol) were used to obtain Synthesis Example 19 as a white solid (40 mg, 14.0%).

[0180] .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ 3.85 (s, 3H), 4.77 (s, 2H), 4.96 (s, 2H), 7.22-7.26 (m, 2H), 7.27-7.28 (d, J=8.0 Hz, 2H), 7.32 (m, 2H), 7.45-7.46 (m, 2H), 7.93 (d, J=8.0 Hz, 2H), 8.22 (br s, 1H).

[Example 4] Confirmation of Degree of Inhibition of HAT p300 Activity of Synthesis Examples 1 to 19

[0181] Synthesis Examples 1 to 19 were diluted to 100 μM, and the degree of inhibition of HAT p300 activity was then determined in the same manner as in Section [2-1]. The results are shown in FIG. 9. Here, the tissues were treated with C646 and Candidate 12 (HAT-12) as the control, instead of Synthesis Examples 1 to 19.

[0182] As shown in FIG. 9, it was confirmed that the HAT p300 activity inhibitory effects of Synthesis Examples 1 to 19 (A 1 to A 19) were 50% on average. In particular, Synthesis Example 6 to 8 had a HAT p300 activity inhibitory effect of approximately 80%, which was nearly similar to that of Candidate 12 (HAT 12). In addition, 90% or more of the HAT p300 activity was inhibited by Synthesis Example 19 (A 19).

[0183] Based on the results, it can be seen that the novel synthetic compound according to the present invention can more effectively inhibit HAT p300 activity because the novel synthetic compound is modified so that additional hydrogen bonds were formed with R1410, T1411, W1466, and Y1467 of HAT p300. From the results, it can be seen that the novel synthetic compound according to the present invention can be very effectively used to prevent, ameliorate or treat diseases associated with the HAT p300.

[0184] While the present invention has been described in detail with reference to exemplary embodiments of the present invention, it will be obvious to those of ordinary skill in the art that this specific description is just a preferred embodiment, and is not intended to limit the scope of the present invention. Therefore, it should be understood that the technical scope of the present invention should be defined by the appended claims and equivalents thereof.

INDUSTRIAL APPLICABILITY

[0185] The present invention relates to a novel compound which enables additional hydrogen bonding with a specific amino acid position of histone acetyltransferase (HA T) p300 through the structural analysis of the HAT p300. The novel compound of the present invention has an excellent inhibitory effect on HAT p300 activity, and thus can be very effectively used to prevent, ameliorate or treat a disease associated with the activation of HAT p300, for example, fibrosis.