KR20220052435A - Aminopyridine derivatives pharmaceutical composition having Bruton's Tyrosine kinase inhibitory activity - Google Patents

Abstract

The present invention relates to an aminopyridine-based compound and a pharmaceutical composition for preventing or treating BTK-related diseases containing the same. The aminopyridine-based compound of the present invention not only exhibits excellent binding ability to a BTK active site, but also has high specificity and selectivity for BTK, so that there is a useful effect of providing the pharmaceutical composition which can lower side effects with the excellent binding ability and high specificity and selectivity and achieve improved medical efficacy in the prevention or treatment of BTK-associated diseases.

Description

Aminopyridine derivatives pharmaceutical composition having Bruton's Tyrosine kinase inhibitory activity

It relates to a pharmaceutical composition having an aminopyridine-based Bruton's tyrosine kinase inhibitory activity.

One of tyrosine protein kinase Tec (Tyrosine protein kinase Tec, TEC), Bruton's tyrosine kinase (BTK), is expressed in B cells, myeloid cells, mast cells, and macrophages. BTK plays an important role in the regulation of B cell growth and function, and is activated downstream of the B cell receptor (BCR). Abnormal function of BTK plays an important role in multiple stages of B cell proliferation, growth, differentiation and apoptosis, leading to hypersensitivity inflammation and hematologic malignancy (Buggy, J. J. et al., International reviews of immunology 2012). , 31(2), 119-32).

Ibrutinib is the first FDA-approved BTK inhibitor, an irreversible covalent inhibitor that targets Cys481, a residue adjacent to the ATP binding site. Ibrutinib is indicated for the treatment of chronic lymphocytic leukemia, Waldenstrom macroglobulinemia, mantle cell lymphoma and chronic graft versus host disease. Approved. Another irreversible BTK inhibitor, acalabrutinib, has received FDA approval for the treatment of mantle cell lymphoma, and several other covalent inhibitors are in clinical trials (Evobrutinib, RN-486, CHMFL). -BTK-01). Most side effects of ibrutinib are expected to be related to off-target effects due to TEC, JAK3 and EGFR inhibition. They can be classified into Group 3F according to the location of the Cys residue near the ATP binding site. In order to overcome these side effects, it is necessary to discover highly selective BTK inhibitors.

Ibrutinib contains a phenoxyphenyl group that forms a hydrophobic interaction with the selectivity pocket formed by Met449, Ile472, Leu542 and Phe540. The selectivity pocket lies behind the Thr474 gatekeeper residue and is driven by the αC-helix out conformation. Acalabrutinib has a pyridylamide group instead of a phenoxy group as this lipophilic substituent. This specific conformational change results in a greatly improved kinase selectivity profile, indicating that changing the lipophilic group is a good approach for BTK selectivity. Through this, the present inventors predicted that by extending the lipophilic linker, not only the αC-helix out conformation but also the DFG-out conformation, which can increase the affinity and selectivity of the compound, could be induced. Since there are currently no effective type II BTK inhibitors, the design of type II BTK inhibitors will be a good starting point for the development of novel selective BTK inhibitors.

Under the above background, the present inventors have completed the present invention by confirming that aminopyridine derivatives exhibit high selectivity to the BTK active site while trying to develop a covalent BTK inhibitor with high selectivity.

One object of the present invention is to provide a BTK inhibitor capable of excellent binding to the BTK active site and at the same time having high specificity and selectivity for BTK, thereby reducing the risk of side effects and achieving improved drug efficacy. To provide a pharmaceutical composition for preventing or treating BTK-related diseases.

In order to achieve the above object,

Provided are a compound represented by the following formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[Formula 1]

In Formula 1,

또는

이고;wherein X ₁ is

or

ego;

wherein R ₂ is hydrogen or —OR ₃ ;

Wherein R ₁ and R ₃ are each independently hydrogen, a substituted or unsubstituted straight _- chain or branched C _{1-10 alkyl} , or a substituted or unsubstituted C _1-10 alkoxycarbonyl,

wherein said substituted straight _- chain or branched C _{1-10 alkyl} , and C _1-10 alkoxy are each halogen, phenyl unsubstituted or substituted with one or more C _1-5 alkoxy or halogen, _and C _{3-8 cycloalkyl} substituted with one or more substituents selected from the group consisting of; and

이다.wherein X ₂ is

am.

Another aspect of the present invention provides a pharmaceutical composition for preventing or treating a BTK (Bruton's tyrosine kinase)-related disease comprising the compound represented by Formula 1, a stereoisomer, or a pharmaceutically acceptable salt thereof as an active ingredient do.

Another aspect of the present invention is a health functional food composition for preventing or improving BTK (Bruton's tyrosine kinase)-related disease containing the compound represented by Formula 1, a stereoisomer, or a pharmaceutically acceptable salt thereof as an active ingredient provides

Another aspect of the present invention is a BTK (Bruton's tyrosine kinase)-related disease comprising administering to a subject in need thereof a compound represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof treatment methods are provided.

In another aspect of the present invention, in the manufacture of a drug for use in the prevention or treatment of BTK (Bruton's tyrosine kinase)-related diseases, the use of the compound represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof provides

The aminopyridine-based compound of the present invention not only exhibits excellent binding to the BTK active site, but also has high specificity and selectivity for BTK, thereby lowering concerns about side effects and achieving improved efficacy in preventing or treating BTK-related diseases. There is a useful effect provided that a pharmaceutical composition capable of doing so.

1 is a diagram showing the prediction of the binding mode of the compound of Example 16 to the kinase domain of BTK as a result of docking analysis of the compound of Example 16 (green line indicates hydrogen bonding interaction).
2 is a diagram showing a hydrophobicity map of the active site as a result of docking analysis of the compound of Example 16 (purple line indicates alkyl-alkyl interaction).
3 shows the compounds of Example 9 and Example 16. It is a diagram showing the change in body weight as a result of the analysis of the in vivo xenograft model.
4 shows the compounds of Example 9 and Example 16. As a result of the analysis of the in vivo xenograft model, it is a diagram showing the change in the volume of the tumor as a graph.

Hereinafter, the present invention will be described in detail.

One aspect of the present invention provides a compound represented by the following formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[Formula 1]

In Formula 1,

또는

이고;wherein X ₁ is

or

ego;

wherein R ₂ is hydrogen or —OR ₃ ;

Wherein R ₁ and R ₃ are each independently hydrogen, a substituted or unsubstituted straight _- chain or branched C _{1-10 alkyl} , or a substituted or unsubstituted C _1-10 alkoxycarbonyl,

wherein said substituted straight _- chain or branched C _{1-10 alkyl} , and C _1-10 alkoxy are each halogen, phenyl unsubstituted or substituted with one or more C _1-5 alkoxy or halogen, _and C _{3-8 cycloalkyl} substituted with one or more substituents selected from the group consisting of; and

이다.wherein X ₂ is

am.

In one embodiment, R ₁ and R ₃ are each independently hydrogen, a substituted or unsubstituted straight _- chain or branched C _{1-6 alkyl} , or a substituted or unsubstituted C _1-6 alkoxycarbonyl,

wherein said substituted straight _{- chain} or branched C _{1-6 alkyl} , and C _1-6 alkoxy are each halogen, phenyl unsubstituted or substituted with one or more C _1-3 alkoxy or halogen, and C _{3-6 cycloalkyl} It is substituted with one or more substituents selected from the group consisting of.

In another embodiment, R ₁ and R ₃ are each independently hydrogen, a substituted or unsubstituted straight-chain or branched C _1-3 alkyl, or a substituted or unsubstituted C _1-3 alkoxycarbonyl,

wherein said substituted straight _- chain or branched C _{1-3 alkyl} , and _{C 1-3 alkoxy} are each halogen, phenyl unsubstituted or substituted _{with one or more C 1-2 alkoxy} or halogen, and C _3-5 cycloalkyl It is substituted with one or more substituents selected from the group consisting of.

자리에 치환된다.In one embodiment, R ₂ is

replaced in place

In one embodiment, the X ₂ is substituted at an ortho or meta site.

,

,

,

,

,

,

,

, 또는

이다.In one embodiment, the R ₁ and R ₃ are each independently hydrogen,

,

,

,

,

,

,

,

, or

am.

In another embodiment, the compound represented by Formula 1 is a compound, characterized in that any one selected from the following compound group.

(1) acrylic acid 3-(6-amino-5-piperazinylpyridin-3-yloxy)phenyl ester;

(2) acrylic acid 3-[6-amino-5-(4-benzylpiperazinyl)pyridin-3-yloxy]phenyl ester;

(3) acrylic acid 3-{6-amino-5-[4-(4-fluorobenzyl)piperazinyl]pyridin-3-yloxy}phenyl ester;

(4) acrylic acid 3-{6-amino-5-[4-(4-chlorobenzyl)piperazinyl]pyridin-3-yloxy}phenyl ester;

(5) acrylic acid 3-[6-amino-5-(4-phenethylpiperazinyl)pyridin-3-yloxy]phenyl ester;

(6) acrylic acid 3-(6-amino-5-{4-[2-(4-chlorophenyl)ethyl]piperazinyl}pyridin-3-yloxy)phenyl ester;

(7) 4-[5-(3-acryloyloxyphenoxy)-2-aminopyridin-3-yl]piperazine carboxylic acid benzylester;

(8) acrylic acid 2-[6-amino-5-(3,4-dihydro-1H-isoquinolin-2-yl)pyridin-3-yloxy]phenyl ester;

(9) acrylic acid 2-[6-amino-5-(6-benzyloxy-3,4-dihydro-1H-isoquinolin-2-yl)pyridin-3-yloxy]phenyl ester;

(10) acrylic acid 3-[6-amino-5-(3,4-dihydro-1H-isoquinolin-2-yl)pyridin-3-yloxy]phenylester;

(11) acrylic acid 3-[6-amino-5-(6-propooxy-3,4-dihydro-1H-isoquinolin-2-yl)pyridin-3-yloxyphenyl ester;

(12) acrylic acid 3-[6-amino-5-(6-cyclopropylmethoxy-3,4-dihydro-1H-isoquinolin-2-yl)pyridin-3-yloxy]phenyl ester;

(13) acrylic acid 3-[6-amino-5-(6-benzyloxy-3,4-dihydro-1H-isoquinolin-2-yl)pyridin-3-yloxy]phenylester;

(14) acrylic acid 3-{6-amino-5-[6-(4-fluorobenzyloxy)-3,4-dihydro-1H-isoquinolin-2-yl]pyridin-3-yloxy}phenyl ester ;

(15) acrylic acid 3-{6-amino-5-[6-(4-chlorobenzyloxy)-3,4-dihydro-1H-isoquinolin-2-yl]pyridin-3-yloxy}phenyl ester; and

(16) acrylic acid 3-{6-amino-5-[6-(4-methoxybenzyloxy)-3,4-dihydro-1H-isoquinolin-2-yl]pyridin-3-yloxy}phenyl ester .

The compound represented by Formula 1 of the present invention may be used in the form of a pharmaceutically acceptable salt, and as the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid, etc., aliphatic mono and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkanes. Non-toxic organic acids such as dioates, aromatic acids, aliphatic and aromatic sulfonic acids, etc., organic acids such as trifluoroacetic acid, acetate, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, fumaric acid, etc. get it from Examples of such pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, and Odide, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, sube Late, sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, Methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, βhydroxybutyrate, glycol late, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate, and the like.

The acid addition salt according to the present invention can be prepared by a conventional method, for example, a precipitate formed by dissolving the derivative of Formula 1 in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile, etc. and adding an organic or inorganic acid It can be prepared by filtration and drying, or by distilling the solvent and excess acid under reduced pressure, followed by drying and crystallization in an organic solvent.

In addition, a pharmaceutically acceptable metal salt can be prepared using a base. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and evaporating and drying the filtrate. In this case, it is pharmaceutically suitable to prepare a sodium, potassium or calcium salt as the metal salt. The corresponding salt is also obtained by reacting an alkali metal or alkaline earth metal salt with a suitable negative salt (eg silver nitrate).

Furthermore, the present invention includes not only the compound represented by Formula 1 and pharmaceutically acceptable salts thereof, but also solvates, optical isomers, hydrates, and the like, which may be prepared therefrom.

Another aspect of the present invention provides a pharmaceutical composition for preventing or treating a BTK (Bruton's tyrosine kinase)-related disease comprising the compound represented by Formula 1, a stereoisomer, or a pharmaceutically acceptable salt thereof as an active ingredient do.

The BTK-related disease may be understood as a disease caused or caused by aberrant activity or expression of BTK, for example, overexpression of BTK, or may be understood to include diseases reported to be related to BTK to date, e.g. For example, the BTK-related disease may be an autoimmune disease or cancer.

In one embodiment, the BTK-related disease is hematological malignancy, hematologic cancer, B-cell chronic lymphocytic leukemia, acute lymphoblastic leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma, acute myeloid leukemia, diffuse giant B -Can be cell lymphoma, multiple myeloma, jacket cell lymphoma, small lymphocytic lymphoma, mantle cell lymphoma (MCL) or lymphocytic leukemia (CLL).

In another embodiment, the BTK-related disease is rheumatoid arthritis, osteoarthritis, juvenile arthritis, chronic obstructive pulmonary disease, multiple sclerosis, asthma, systemic lupus erythematosus, psoriasis, psoriatic arthritis, Crohn's disease, ulcerative colitis or irritable bowel syndrome can be

The compound of formula 1 of the present invention is a small molecule compound that binds to the BTK active site, and can form an irreversible covalent bond with the BTK active site. Accordingly, although the present invention is not limited to a particular theory, for example, the compound of Formula 1 of the present invention is an irreversible inhibitor of BTK, which inhibits BTK activity to prevent or treat BTK-related diseases. In addition, the compound of formula 1 of the present invention is a compound with high specificity and selectivity for BTK, and there is little concern about side effects, so that the drug can be used at a higher dose and sufficient drug efficacy can be achieved for disease treatment.

The compound of formula 1 of the present invention binds to the BTK active site through hydrogen bonding and van der Waals bonding, thereby increasing specificity and selectivity for BTK (see FIGS. 1 and 2 ). In one embodiment of the present invention, it was confirmed that the aminopyridine-based compound of Formula 1 of the present invention had high specificity and selectivity for BTK, and significantly reduced tumor size without weight loss in an in vivo xenograft model (Fig. 3 and 4).

The pharmaceutical composition of the present invention may be used as a single agent, and may be prepared and used as a combination formulation by additionally including a recognized BTK inhibitor or a pharmaceutical composition known to have a preventive or therapeutic effect on BTK-related diseases. A pharmaceutical unit dosage form may be formulated by adding a pharmaceutically acceptable carrier, excipient, or diluent.

In the present invention, "treatment" refers to any action in which the symptoms of BTK-related diseases are improved or beneficially changed by administration of the composition according to the present invention.

In the present invention, "prevention" refers to any action that inhibits or delays the onset of BTK-related diseases by administration of the composition according to the present invention.

In the present invention, "pharmaceutically acceptable" means that it does not significantly stimulate the organism and does not impair the biological activity and properties of the administered active substance.

In the present invention, the pharmaceutical composition comprising a pharmaceutically acceptable carrier is a tablet, pill, powder, granule, capsule, suspension, internal solution, emulsion, syrup, sterile aqueous solution, non-aqueous solvent, suspension, emulsion, It may have any one formulation selected from the group consisting of freeze-dried formulations and suppositories. The pharmaceutical composition may be in various oral or parenteral formulations. In the case of formulation, it can be prepared using a diluent or excipient such as a filler, extender, binder, wetting agent, disintegrant, surfactant, etc. commonly used.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations include at least one excipient in one or more compounds, for example, starch, calcium carbonate, sucrose or lactose ( lactose), gelatin, etc. can be mixed to prepare it. In addition to simple excipients, lubricants such as magnesium stearate, talc and the like may also be used. Liquid preparations for oral administration include suspensions, internal solutions, emulsions, syrups, etc. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. there is.

Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized formulations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, etc. may be used, but is not limited thereto.

In the composition of the present invention, the compound of Formula 1 may be included in a pharmaceutically effective amount. "Pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment and not to cause side effects, and the effective dose level is determined by the patient's health condition, type of disease, The severity, activity of the drug, sensitivity to the drug, administration method, administration time, administration route and excretion rate, duration of treatment, factors including drugs used in combination or concomitantly, and other factors well known in the medical field can be determined according to factors. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiple times. In consideration of all of the above factors, it is important to administer an amount that can obtain the maximum effect with a minimum amount without side effects, which can be easily determined by those skilled in the art.

Another aspect of the present invention is a health functional food composition for preventing or improving BTK (Bruton's tyrosine kinase)-related disease containing the compound represented by Formula 1, a stereoisomer, or a pharmaceutically acceptable salt thereof as an active ingredient provides

In the present invention, "improvement" refers to any action that reduces a parameter related to a condition to be treated by ingestion of the composition, for example, the severity of symptoms.

The health food refers to food manufactured and processed in the form of tablets, capsules, powders, granules, liquids and pills using raw materials or ingredients useful for the human body. The health functional food of the present invention can be prepared by a method commonly used in the art, and during the manufacture, it can be prepared by adding raw materials and components commonly added in the art. In addition, the dosage form of the health functional food may be manufactured without limitation as long as it is a dosage form recognized as a health functional food.

Another aspect of the present invention is a BTK (Bruton's tyrosine kinase)-related disease comprising administering to a subject in need thereof a compound represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof treatment methods are provided.

In the present invention, the term "subject" includes both mammals and non-mammals that have or are likely to develop the BTK-related disease, for example, humans.

In the present invention, "administration" means providing a predetermined substance to a patient by any suitable method, and the administration route of the composition of the present invention may be administered through any general route as long as it can reach the target tissue. there is.

In another aspect of the present invention, in the manufacture of a drug for use in the prevention or treatment of BTK (Bruton's tyrosine kinase)-related diseases, the use of the compound represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof provides

Hereinafter, Examples and Experimental Examples of the present invention will be specifically illustrated and described below. However, the Examples and Experimental Examples to be described below are only illustrative of a part of the present invention, and are not limited thereto.

< Example 1> Acrylic acid 3-(6-amino-5- piperazinylpyridine -3- Iloxy ) Preparation of phenyl esters

[Scheme 1]

As in Reaction Scheme 1, the amine group of 2-amino-3,5-dichloropyridine was converted to a nitro group by reacting under the conditions of K ₂ S ₂ 0 ₈ , H ₂ SO ₄ and 90 ° C. for 24 hours. , 1-Boc-piperazine, K ₂ CO ₃ , toluene, and at 50 ° C. for 15 hours to replace the C3 position of pyridine with Boc-piperazine, and then deprotect the Boc group with TFA. Thus, 3-piperazinylpyridine was prepared. Then, Cs ₂ CO ₃ in a DMF solvent was reacted at 70 ° C. for 24 hours to introduce a resorcinol group through nucleophilic substitution, followed by acylation with a Michael acceptor, and SnCl ₂ ㆍ2H ₂ Acrylic acid 3-(6-amino-5-piperazinylpyridin-3-yloxy)phenyl ester (Acrylic acid 3-( 6-amino-5-piperazinylpyridin-3-yloxy)phenyl ester) was prepared.

brown solid (yield 4%); IR (neat, cm ^-1 ) 3340.09, 2923.62, 2852.31, 1735.98, 1640.23, 1595.58, 1466.46, 1367.40, 1335.50, 1272.67, 1244.25, 1176.36, 1137.91, 1043.36, 993.22, 967.52; ¹ H NMR (500 MHz, CDCl ₃ ) δ 7.63 (1H, br s), 7.14 (1H, t, J = 8.0 Hz), 6.96 (1H, d, J = 2.5 Hz), 6.58 (1H, dd, J = 10.5, 17.0 Hz), 6.55 (1H, dd, J = 2.0, 8.0 Hz), 6.49 (1H, dd, J = 2.0, 8.0 Hz), 6.38 (1H, t, J = 2.0 Hz), 6.33 (1H) , dd, J = 1.5, 17.0 Hz), 5.75 (1H, dd, J = 1.5, 10.5 Hz), 4.90 (2H, br s), 3.81 (2H, br s), 3.71 (2H, br s), 2.90 (4H, br s); ¹³ C NMR (500 MHz, CDCl ₃ ) δ 165.8, 159.7, 157.8, 151.0, 150.2, 134.5, 133.7, 130.7, 128.9, 127.2, 121.1, 110.3, 109.2, 104.4, 51.0, 50.5, 46.3, 42.5; LCMS (ESI) m/z calcd for C ₁₈ H ₂₀ N ₄ O ₃ , 340.15; Found 341.5 [M + H] ⁺ .

< Example 2> Acrylic acid 3-[6-amino-5-(4- benzylpiperazinyl )pyridine-3- Iloxy ]Production of phenyl esters

It was carried out in the same manner as in Example 1, except that after preparing 3-piperazinylpyridine, (halomethyl)benzene, K ₂ at room temperature for N-alkylation of the piperazine group for 5 hours CO ₃ , further comprising the step of reacting with DMF, acrylic acid 3-[6-amino-5-(4-benzylpiperazinyl)pyridin-3-yloxy]phenyl ester (Acrylic acid 3-[6-amino-5 -(4-benzylpiperazinyl)pyridin-3-yloxy]phenyl ester) was prepared.

brown solid (yield 15%); IR (neat, cm ^-1 ) 3058.17, 2942.12, 2820.72, 1664.56, 1594.22, 1462.81, 1369.10, 1336.85, 1265.54, 1214.68, 1178.91, 1134.30, 996.06, 737.15, 700.62; ¹ H NMR (500 MHz, CDCl ₃ ) δ 7.69 (1H, d, J = 2.5 Hz), 7.34-7.26 (6H, m), 6.98 (1H, d, J = 2.5 Hz), 6.83-6.80 (2H, m), 6.70 (1H, t, J = 2.0 Hz), 6.58 (1H, dd, J = 1.0, 17.5 Hz), 6.28 (1H, dd, J = 10.5, 17.5 Hz), 6.00 (1H, dd, J ) = 1.0, 10.5 Hz), 4.63 (2H, br s), 3.58 (2H, s), 2.93 (4H, br s), 2.60 (4H, br s); ¹³ C NMR (500 MHz, CDCl ₃ ) δ 164.3, 159.7, 151.6, 151.4, 145.1, 138.0, 135.2, 134.3, 132.8, 130.1, 129.3, 128.4, 127.9, 127.3, 120.1, 115.6, 114.2, 110.4, 63.1, 53.5 , 50.6; LCMS (ESI) m/z calcd for C ₂₅ H ₂₆ N ₄ O ₃ , 430.20; Found 431.7 [M + H] ⁺ .

< Example 3> Acrylic acid 3-{6-amino-5-[4-(4-) Fluorobenzyl ) Preparation of piperazinyl]pyridin-3-yloxy}phenyl ester

Acrylic acid 3-{6-amino-5-[4-(4-) was carried out in the same manner as in Example 2, but using 1-(halomethyl)-4-fluorobenzene instead of (halomethyl)benzene Preparation of fluorobenzyl)piperazinyl]pyridin-3-yloxy}phenyl ester (Acrylic acid 3-{6-amino-5-[4-(4-fluorobenzyl)piperazinyl]pyridin-3-yloxy}phenyl ester) did

brown solid (yield 12%); IR (neat, cm ^-1 ) 2939.63, 2821.27, 1740.60, 1602.75, 1507.57, 1479.89, 1402.43, 1366.12, 1312.43, 1243.62, 1220.03, 1157.24, 1025.69, 1002.28, 841.48, 796.97, 736.57, 700.72; ¹ H NMR (500 MHz, CDCl ₃ ) δ 7.68 (1H, d, J = 2.5 Hz), 7.31-7.27 (3H, m), 7.01 (2H, t, J = 8.5 Hz), 6.98 (1H, d, J = 2.5 Hz), 6.81 (2H, t, J = 8.5 Hz), 6.70 (1H, t, J = 2.5 Hz), 6.58 (1H, dd, J = 1.0, 17.5 Hz), 6.28 (1H, dd, J = 10.5, 17.5 Hz), 6.00 (1H, dd, J = 1.0, 10.5 Hz), 4.71 (2H, br s), 3.53 (2H, s), 2.93 (4H, br s), 2.58 (4H, br) s); ¹³ C NMR (500 MHz, CDCl ₃ ) δ 164.3, 162.2 (d, J _{C -F} = 243.9 Hz), 159.7, 151.7, 151.5, 145.1, 135.3, 134.2, 133.7 (d, J _{C -F} = 3.1 Hz), 132.9, 130.7 (d, J _{C -F} = 7.9 Hz), 130.2, 127.9, 120.3, 115.6, 115.2 (d, J _{C -F} = 21.1 Hz), 114.2, 110.4, 62.3, 53.5, 50.6; LCMS (ESI) m/z calcd for C ₂₅ H ₂₅ FN ₄ O ₃ , 448.19; Found 449.6 [M + H] ⁺ .

< Example 4> Acrylic acid 3-{6-amino-5-[4-(4- chlorobenzyl ) Preparation of piperazinyl]pyridin-3-yloxy}phenyl ester

It was carried out in the same manner as in Example 2, except that 1-(halomethyl)-4-chlorobenzene was used instead of (halomethyl)benzene, and acrylic acid 3-{6-amino-5-[4-(4-chlorobenzene) was used. Benzyl)piperazinyl]pyridin-3-yloxy}phenyl ester (Acrylic acid 3-{6-amino-5-[4-(4-chlorobenzyl)piperazinyl]pyridin-3-yloxy}phenyl ester) was prepared.

brown solid (10% yield); IR (neat, cm ^-1 ) 3048.82, 2940.26, 2821.49, 1742.76, 1594.96, 1488.78, 1402.63, 1367.11, 1311.85, 1293.49, 1247.75, 1212.80, 1157.75, 1086.29, 1002.35, 795.38, 736.3; ¹ H NMR (500 MHz, CDCl ₃ ) δ 7.69 (1H, d, J = 2.5 Hz), 7.32-7.26 (5H, m), 6.98 (1H, d, J = 2.5 Hz), 6.83-6.80 (2H, m), 6.70 (1H, t, J = 2.5 Hz), 6.58 (1H, dd, J = 1.0, 17.5 Hz), 6.28 (1H, dd, J = 10.5, 17.5 Hz), 6.00 (1H, dd, J ) = 1.0, 10.5 Hz), 4.63 (2H, br s), 3.53 (2H, s), 2.93 (4H, br s), 2.58 (4H, br s); ¹³ C NMR (500 MHz, CDCl ₃ ) δ 164.4, 159.8, 151.7, 151.5, 145.1, 136.6, 135.2, 134.5, 133.0, 132.9, 130.5, 130.2, 128.6, 127.9, 120.2, 115.6, 114.2, 110.4, 62.3, 53.5 , 50.6; LCMS (ESI) m/z calcd for C ₂₅ H ₂₅ ClN ₄ O ₃ , 464.16; Found 465.6 [M + H] ⁺ .

< Example 5> Acrylic acid 3-[6-amino-5-(4- phenethylpiperazinyl )pyridine-3- Iloxy ]Production of phenyl esters

It was carried out in the same manner as in Example 2, except that (2-haloethyl)benzene was used instead of (halomethyl)benzene, and acrylic acid 3-[6-amino-5-(4-phenethylpiperazinyl)pyridine- 3-yloxy] phenyl ester (Acrylic acid 3- [6-amino-5- (4-phenethylpiperazinyl) pyridin-3-yloxy] phenyl ester) was prepared.

brown solid (yield 19%); IR (neat, cm ^-1 ) 3025.91, 2942.72, 2819.03, 1743.57, 1608.10, 1464.13, 1402.22, 1372.06, 1336.13, 1247.66, 1152.02, 1023.95, 1001.77, 775.70, 735.23, 700.10; ¹ H NMR (500 MHz, CDCl ₃ ) δ 7.70 (1H, d, J = 2.5 Hz), 7.31-7.28 (3H, m), 7.23-7.21 (3H, m), 7.00 (1H, d, J = 2.5) Hz), 6.83-6.81 (2H, m), 6.71 (1H, t, J = 2.5 Hz), 6.59 (1H, dd, J = 1.0, 17.5 Hz), 6.29 (1H, dd, J = 10.5, 17.5 Hz) ), 6.01 (1H, dd, J = 1.0, 10.5 Hz), 4.65 (2H, br s), 2.97 (4H, br s), 2.85-2.82 (2H, m), 2.69-2.66 (6H, m); ¹³ C NMR (500 MHz, CDCl ₃ ) δ 164.4, 159.7, 151.7, 151.4, 145.1, 140.3, 135.2, 134.4, 132.9, 130.2, 128.8, 128.6, 127.9, 126.3, 120.2, 115.6, 114.2, 110.4, 60.6, 53.7 , 50.6, 33.8; LCMS (ESI) m/z calcd for C ₂₆ H ₂₈ N ₄ O ₃ , 444.22; Found 445.6 [M + H] ⁺ .

< Example 6> Acrylic acid 3-(6-amino-5-{4-[2-(4- chlorophenyl Preparation of )ethyl]piperazinyl}pyridin-3-yloxy)phenyl ester

Acrylic acid 3-(6-amino-5-{4-[2] was carried out in the same manner as in Example 2, except that 1-chloro-4-(2-haloethyl)benzene was used instead of (halomethyl)benzene. -(4-chlorophenyl)ethyl]piperazinyl}pyridin-3-yloxy)phenyl ester (Acrylic acid 3-(6-amino-5-{4-[2-(4-chlorophenyl)ethyl]piperazinyl}pyridin) -3-yloxy)phenyl ester) was prepared.

brown solid (yield 4%); IR (neat, cm ^-1 ) 3355.04, 2938.20, 2821.01, 1742.91, 1608.93, 1464.96, 1402.98, 1336.85, 1247.88, 1154.36, 1002.08, 776.31; ¹ H NMR (500 MHz, CDCl ₃ ) δ 7.69 (1H, d, J = 2.5 Hz), 7.29 (1H, t, J = 8.5 Hz), 7.25 (2H, d, J = 8.5 Hz), 7.14 (2H) , d, J = 8.5 Hz), 6.99 (2H, d, J = 2.5 Hz), 6.83-6.80 (2H, m), 6.71 (1H, t, J = 2.5 Hz), 6.58 (1H, dd, J = 2.5 Hz) 1.0, 17.5 Hz), 6.29 (1H, dd, J = 10.5, 17.5 Hz), 6.01 (1H, dd, J = 1.0, 10.5 Hz), 4.68 (2H, br s), 2.96 (4H, br s), 2.81-2.78 (2H, m), 2.65-2.62 (6H, m); LCMS (ESI) m/z calcd for C ₂₆ H ₂₇ ClN ₄ , 478.18; Found 479.7 [M + H] ⁺ .

< Example 7> 4-[5-(3- acryloyloxyphenoxy )-2- aminopyridine -3-yl]piperazine carboxylic acid of benzyl ester Produce

It was carried out in the same manner as in Example 2, but using benzyl carbonohaloridate instead of (halomethyl)benzene, 4-[5-(3-acryloyloxyphenoxy)-2-aminopyridine-3 -yl]piperazine carboxylic acid benzyl ester (4-[5-(3-Acryloyloxyphenoxy)-2-aminopyridin-3-yl]piperazine carboxylic acid benzyl ester) was prepared.

brown solid (23% yield); IR (neat, cm ^-1 ) 3351.96, 3062.09, 2827.34, 1742.87, 1688.0, 1608.87, 1466.27, 1403.28, 1361.16, 1333.67, 1284.80, 1245.40, 1152.18, 1087.69. 1044.80, 992.50, 908.43, 735.68, 699.05; ¹ H NMR (500 MHz, CDCl ₃ ) δ 7.72 (1H, d, J = 2.5 Hz), 7.39-7.32 (5H, m), 7.29 (1H, t, J = 8.0 Hz), 6.95 (1H, d, J = 2.5 Hz), 6.82 (2H, td, J = 2.0, 8.0 Hz), 6.70 (1H, t, J = 2.5 Hz), 6.58 (1H, dd, J = 1.0, 17.5 Hz), 6.28 (1H, dd, J = 10.5, 17.5 Hz), 6.01 (1H, dd, J = 1.0, 10.5 Hz), 5.16 (2H, s), 4.65 (2H, br s), 3.65 (4H, br s), 2.88 (4H) , br s); ¹³ C NMR (500 MHz, CDCl ₃ ) δ 164.3, 159.6, 155.4, 151.7, 151.3, 145.2, 136.7, 135.1, 134.6, 133.0, 130.3, 128.7, 128.3, 128.1, 127.9, 120.4, 115.7, 114.3, 110.5, 67.5 , 50.5, 44.4; LCMS (ESI) m/z calcd for C ₂₆ H ₂₆ N ₄ O ₅ , 474.19; Found 475.7 [M + H] ⁺ .

< Example 8> Acrylic acid 2- [6-amino-5- (3,4- dihydro -1H- isoquinoline Preparation of -2-yl) pyridin-3-yloxy] phenyl ester

[Scheme 2]

As in Scheme 2, 3,5-dichloro-2-nitropyridine was reacted with K ₂ CO ₃ , toluene, tetrahydroisoquinoline for 10 hours at 60 ° C. conditions, K ₂ CO ₃ , DMSO, 18-crown-6, reacted with catechol for 10 hours at 100 ° C., and reacted with acryloyl chloride for 1 hour under TEA and DCM conditions at room temperature. Then, SnCl ₂ ·2H ₂ O, EtOH was reacted at room temperature for 1 hour to convert the nitro group to an amine, and acrylic acid 2-[6-amino-5-(3,4-dihydro-1H-isoquinoline) -2-yl) pyridin-3-yloxy] phenyl ester (Acrylic acid 2- [6-amino-5- (3,4-dihydro- 1H -isoquinolin-2-yl) pyridin-3-yloxy] phenyl ester) was prepared.

yellow solid (yield 31%); IR (neat, cm ^-1 ) 2980.52, 2888.53, 1596.48, 1460.26, 1380.12, 1315.59, 1230.72, 1315.59, 1238.76, 1150.52, 1023.45, 971.17; ¹ H NMR (500 MHz, CDCl ₃ ) δ 7.69 (1H, d, J = 2.5 Hz), 7.21-7.14 (5H, m), 7.10-7.06 (2H, m), 7.03 (1H, d, J = 2.5) Hz), 6.89 (1H, dd, J = 1.5, 8.5 Hz), 6.59 (1H, dd, J = 1.0, 17.5 Hz), 6.32 (1H, dd, J = 10.5, 17.5 Hz), 6.00 (1H, dd) , J = 1.0, 10.5 Hz), 4.66 (2H, br s), 4.06 (2H, s), 3.22 (2H, t, J = 6.0 Hz), 3.01 (2H, t, J = 6.0 Hz); ¹³ C NMR (500 MHz, CDCl ₃ ) δ 164.1, 151.4, 150.2, 145.8, 140.8, 135.1, 134.5, 133.9, 132.9, 129.1, 127.6, 127.1, 126.6, 123.7, 123.3, 119.9, 118.0, 53.2, 48.9, 29.8 ; LCMS (ESI) m/z calcd for C ₂₃ H ₂₁ N ₃ O ₃ , 387.16; Found 388.16 [M + H] ⁺ .

< Example 9> Acrylic acid 2- [6-amino-5- (6- benzyloxy -3,4- dihydro -1H- isoquinoline Preparation of -2-yl) pyridin-3-yloxy] phenyl ester

Acrylic acid 2-[6-amino-5-] was carried out in the same manner as in Example 8, except that 6-(benzyloxy)-1,2,3,4-tetrahydroisoquinoline was used instead of tetrahydroisoquinoline. (6-Benzyloxy-3,4-dihydro-1H-isoquinolin-2-yl)pyridin-3-yloxy]phenyl ester (Acrylic acid 2-[6-amino-5-(6-benzyloxy-3, 4-dihydro- 1H -isoquinolin-2-yl)pyridin-3-yloxy]phenyl ester) was prepared.

ivory solid (23% yield); IR (neat, cm ^-1 ) 2980.65, 2888.60, 1735.13, 1610.91, 1461.97, 1380.75, 1239.82, 1150.48, 1015.44, 969.35; ¹ H NMR (500 MHz, CDCl ₃ ) δ 7.68 (1H, d, J = 2.5 Hz), 7.42 (2H, d, J = 7.5 Hz), 7.38 (2H, t, J = 7.5 Hz), 7.31 (1H) , t, J = 7.5 Hz), 7.15 (2H, t, J = 7.5 Hz), 7.07 (1H, t, J = 7.5 Hz), 7.01 (1H, d, J = 2.5 Hz), 6.97 (1H, d) , J = 8.5 Hz), 6.88 (1H, d, J = 8.0 Hz), 6.81-6.77 (2H, m), 6.57 (1H, dd, J = 1.0, 17.0 Hz), 6.31 (1H, dd, J = 10.5, 17.0 Hz), 5.98 (1H, dd, J = 1.0, 10.5 Hz), 5.04 (2H, s), 4.72 (2H, br s), 3.98 (2H, s), 3.17 (2H, t, J = 5.5 Hz), 2.95 (2H, t, J = 5.5 Hz); ¹³ C NMR (500 MHz, CDCl ₃ ) δ 164.1, 159.8, 157.6, 154.5, 151.4, 150.2, 145.7, 140.7, 137.2, 135.1, 135.1, 133.7, 132.9, 130.2, 128.7, 128.1, 127.5, 127.5, 127.1, 126.9 , 123.6, 123.2, 119.8, 117.9, 114.7, 113.2, 70.2, 52.7, 48.7, 30.0; LCMS (ESI) m/z calcd for C ₃₀ H ₂₇ N ₃ O ₄ , 493.20; Found 494.20 [M + H] ⁺ .

< Example 10> Acrylic acid 3- [6-amino-5- (3,4- dihydro -1H- isoquinoline Preparation of -2-yl) pyridin-3-yloxy] phenyl ester

It was carried out in the same manner as in Example 8, except that resorcinol was used instead of catechol and acrylic acid 3-[6-amino-5-(3,4-dihydro-1H-isoquinolin-2-yl)pyridine was used. -3-yloxy]phenyl ester (Acrylic acid 3-[6-amino-5-(3,4-dihydro- 1H -isoquinolin-2-yl)pyridin-3-yloxy]phenyl ester) was prepared.

yellow solid (yield 30%); IR (neat, cm ^-1 ) 2980.40, 2888.74, 1461.83, 1382.18, 1239.58, 1151.98, 966.99; ¹ H NMR (500 MHz, CDCl ₃ ) δ 7.73 (1H, d, J = 2.5 Hz), 7.30 (1H, t, J = 8.0 Hz), 7.21-7.15 (3H, m), 7.07 (1H, d, J = 7.0 Hz), 7.06 (1H, d, J = 2.5 Hz), 6.83 (1H, dd, J = 2.5, 8.5 Hz), 6.82 (1H, dd, J = 2.0, 8.5 Hz), 6.73 (1H, dd, J = 2.0, 2.5 Hz), 6.59 (1H, d, J = 1.5, 17.5 Hz), 6.29 (1H, dd, J =10.5, 17.5 Hz), 6.01 (1H, d, J = 1.5, 10.5 Hz) ), 4.73 (2H, br s), 4.09 (2H, s), 3.24 (2H, t, J = 6.0 Hz), 3.02 (2H, t, J = 6.0 Hz); ¹³ C NMR (500 MHz, CDCl ₃ ) δ 164.4, 159.7, 151.8, 151.6, 144.9, 135.4, 134.3, 133.8, 133.8, 132.9, 130.2, 129.1, 127.9, 126.7, 126.5, 126.1, 120.9, 115.7, 114.2, 110.4 , 53.3, 48.9, 29.6; LCMS (ESI) m/z calcd for C ₂₃ H ₂₁ N ₃ O ₃ , 387.16; Found 388.16 [M + H] ⁺ .

< Example 11> Acrylic acid 3-[6-amino-5-(6- propoxy -3,4- dihydro Preparation of -1H-isoquinolin-2-yl)pyridin-3-yloxyphenyl ester

Acrylic acid 3-[6-amino-5-(6) was carried out in the same manner as in Example 10, except that 6-propoxy-1,2,3,4-tetrahydroisoquinoline was used instead of tetrahydroisoquinoline. -Propoxy-3,4-dihydro-1H-isoquinolin-2-yl)pyridin-3-yloxyphenyl ester (Acrylic acid 3-[6-amino-5-(6-propoxy-3,4-) dihydro- 1H -isoquinolin-2-yl)pyridin-3-yloxy]phenyl ester) was prepared.

yellow solid (23% yield); IR (neat, cm ⁻¹ ) 2980.97, 2883.67, 1595.77, 1459.04, 1399.69, 1381.50, 1314.25, 1237.69, 1151.35, 1023.03, 981.04; ¹ H NMR (500 MHz, CDCl ₃ ) δ 7.72 (1H, d, J = 2.5 Hz), 7.30 (1H, t, J = 8.5 Hz), 7.05 (1H, d, J = 2.5 Hz), 6.98 (1H) , d, J = 8.5 Hz), 6.83-6.81 (2H, m), 6.75-6.70 (3H, m), 6.58 (1H, dd, J = 1.0, 17.5 Hz), 6.29 (1H, dd, J = 10.5) , 17.5 Hz), 6.00 (1H, dd, J = 1.0, 10.5 Hz), 4.73 (2H, br s), 4.02 (2H, s), 3.91 (2H, t, J = 7.0 Hz), 3.21 (2H, t, J = 6.0 Hz), 2.97 (2H, t, J = 6.0 Hz), 1.80 (2H, sext., J = 7.0 Hz), 1.03 (3H, t, J = 7.0 Hz); ¹³ C NMR (500 MHz, CDCl ₃ ) δ 164.3, 159.6, 157.8, 151.6, 151.5, 144.9, 135.3, 134.9, 133.8, 133.2, 132.8, 130.2, 127.8, 127.3, 126.2, 125.4, 120.9, 115.5, 114.3, 114.1 , 110.2, 52.7, 52.0, 51.5, 48.7, 29.2; LCMS (ESI) m/z calcd for C ₂₆ H ₂₇ N ₃ O ₄ , 445.20; Found 444.21 [M + H] ⁺ .

< Example 12> Acrylic acid 3-[6-amino-5-(6- cyclopropylmethoxy -3,4- dihydro Preparation of -1H-isoquinolin-2-yl)pyridin-3-yloxy]phenyl ester

Acrylic acid 3- [6-amino- 5- (6-cyclopropylmethoxy-3,4-dihydro-1H-isoquinolin-2-yl) pyridin-3-yloxy] phenyl ester (Acrylic acid 3- [6-amino-5- (6- cyclopropylmethoxy-3,4-dihydro- 1H -isoquinolin-2-yl)pyridin-3-yloxy]phenyl ester) was prepared.

yellow solid (yield 67%); IR (neat, cm ^-1 ) 2981.10, 2884.01, 1595.97, 1459.23, 1399.20, 1369.08, 1311.93, 1276.07, 1237.12, 1153.63, 1129.55, 1021.23; ¹ H NMR (500 MHz, CDCl ₃ ) δ 7.70 (1H, d, J = 2.5 Hz), 7.30 (1H, t, J = 8.0 Hz), 7.05 (1H, d, J = 2.5 Hz), 6.98 (1H) , d, J = 8.5 Hz), 6.82 (2H, dd, J = 2.5, 8.5 Hz), 6.76-6.70 (3H, m), 6.59 (1H, dd, J = 1.5, 17.5 Hz), 6.29 (1H, dd, J = 10.5, 17.5 Hz), 6.01 (1H, dd, J = 1.5, 10.5 Hz), 4.79 (2H, br s), 4.02 (2H, s), 3.79 (2H, d, J = 7.0 Hz) , 3.21 (2H, t, J = 6.0 Hz), 2.97 (2H, t, J = 6.0 Hz), 1.30-1.23 (1H, m), 0.64 (2H, dd, J = 5.0, 12.5 Hz), 0.34 ( 2H, dd, J = 5.0, 10.0 Hz); ¹³ C NMR (500 MHz, CDCl ₃ ) δ 164.3, 159.6, 157.7, 151.6, 151.5, 144.9, 135.3, 134.9, 133.9, 132.8, 130.1, 127.8, 127.7, 127.4, 126.4, 120.9, 120.6, 115.5, 114.3, 114.1 , 113.0, 110.3, 52.7, 50.7, 48.6, 40.6, 29.8; LCMS (ESI) m/z calcd for C ₂₇ H ₂₇ N ₃ O ₄ , 457.20; Found 458.22 [M + H] ⁺ .

< Example 13> Acrylic acid 3-[6-amino-5-(6- benzyloxy -3,4- dihydro -1H- isoquinoline Preparation of -2-yl) pyridin-3-yloxy] phenyl ester

Acrylic acid 3-[6-amino-5-] was carried out in the same manner as in Example 10, except that 6-(benzyloxy)-1,2,3,4-tetrahydroisoquinoline was used instead of tetrahydroisoquinoline. (6-benzyloxy-3,4-dihydro-1H-isoquinolin-2-yl) pyridin-3-yloxy] phenyl ester (Acrylic acid 3- [6-amino-5- (6-benzyloxy-3, 4-dihydro- 1H -isoquinolin-2-yl)pyridin-3-yloxy]phenyl ester) was prepared.

yellow solid (yield 63%); IR (neat, cm ^-1 ) 2980.57, 1739.16, 1607.15, 1462.79, 1401.07, 1378.91, 1313.43, 1237.60, 1151.77, 1023.59, 1001.65; ¹ H NMR (500 MHz, CDCl ₃ ) δ 7.72 (1H, d, J = 2.5 Hz), 7.43 (2H, d, J = 7.0 Hz), 7.38 (2H, t, J = 7.5 Hz), 7.33-7.28 (2H, m), 7.04 (1H, d, J = 2.5 Hz), 6.99 (1H, d, J = 8.0 Hz), 6.83-6.78 (4H, m), 6.72 (1H, t, J = 2.5 Hz) , 6.58 (1H, dd, J = 1.0, 17.5 Hz), 6.29 (1H, dd, J = 10.5, 17.5 Hz), 6.00 (1H, dd, J = 1.5, 10.5 Hz), 5.05 (2H, s), 4.72 (2H, br s), 4.02 (2H, s), 3.21 (2H, t, J = 6.0 Hz), 2.97 (2H, t, J = 6.0 Hz); ¹³ C NMR (500 MHz, CDCl ₃ ) δ 164.4, 159.8, 157.6, 154.5, 151.7, 151.6, 145.0, 137.2, 135.2, 135.1, 134.5, 132.9, 130.2, 128.7, 128.1, 127.9, 127.6, 127.5, 126.9, 120.5 , 115.6, 114.8, 114.2, 113.3, 110.4, 70.2, 52.8, 48.7, 29.9; LCMS (ESI) m/z calcd for C ₃₀ H ₂₇ N ₃ O ₄ , 493.20; Found 494.21 [M + H] ⁺ .

< Example 14> Acrylic acid 3-{6-amino-5-[6-(4- Fluorobenzyloxy )-3,4- dihydro Preparation of -1H-isoquinolin-2-yl]pyridin-3-yloxy}phenyl ester

Acrylic acid 3-{ 6-amino-5-[6-(4-fluorobenzyloxy)-3,4-dihydro-1H-isoquinolin-2-yl]pyridin-3-yloxy}phenyl ester (Acrylic acid 3-{6 -amino-5-[6-(4-fluorobenzyloxy)-3,4-dihydro- 1H -isoquinolin-2-yl]pyridin-3-yloxy}phenyl ester) was prepared.

yellow solid (yield 4%); IR (neat, cm ^-1 ) 3321.71, 2922.44, 2850.45, 1735.63, 1603.52, 1508.71, 1460.41, 1400.57, 1379.29, 1235.92, 1151.94, 1022.79; ¹ H NMR (500 MHz, CDCl ₃ ) δ 7.71 (1H, d, J = 2.0 Hz), 7.42 (2H, m), 7.32 (1H, t, J = 8.0 Hz), 7.09 (3H, m), 7.00 (1H, d, J = 8.5 Hz), 6.84 (2H, m), 6.81 (1H, dd, J = 2.0, 8.5 Hz), 6.77 (1H, d, J = 2.0 Hz), 6.73 (1H, d, J = 2.0 Hz), 6.57 (1H, d, J = 17.0 Hz), 6.32 (1H, dd, J = 10.5, 17.0 Hz), 6.02 (1H, d, J = 10.5 Hz), 5.01 (2H, s) , 4.82 (2H, br s), 4.03 (2H, s), 3.23 (2H, t, J = 6.0 Hz), 2.99 (2H, t, J = 6.0 Hz); ¹³ C NMR (500 MHz, CDCl ₃ ) δ 164.4, 159.7, 157.4, 151.8, 151.7, 145.0, 135.4, 132.9, 130.2, 129.4, 129.4, 127.9, 127.6, 127.0, 120.8, 115.9, 115.7, 115.7, 115.5, 115.3 , 114.8, 114.2, 113.3, 110.4, 69.5, 53.6, 52.8, 48.7, 29.9; LCMS (ESI) m/z calcd for C ₃₀ H ₂₅ FN ₃ O ₄ , 511.19; Found 512.19 [M + H] ⁺ .

< Example 15> Acrylic acid 3-{6-amino-5-[6-(4- chlorobenzyloxy )-3,4- dihydro Preparation of -1H-isoquinolin-2-yl]pyridin-3-yloxy}phenyl ester

Acrylic acid 3-{6 was carried out in the same manner as in Example 10, except that 6-((4-chlorobenzyl)oxy)-1,2,3,4-tetrahydroisoquinoline was used instead of tetrahydroisoquinoline. -Amino-5-[6-(4-chlorobenzyloxy)-3,4-dihydro-1H-isoquinolin-2-yl]pyridin-3-yloxy}phenyl ester (Acrylic acid 3-{6-amino -5-[6-(4-chlorobenzyloxy)-3,4-dihydro- 1H -isoquinolin-2-yl]pyridin-3-yloxy}phenyl ester) was prepared.

yellow solid (yield 55%); IR (neat, cm ^-1 ) 2980.88, 2952.96, 2888.90, 1458.99, 1443.76, 1234.19, 1148.64, 1015.34; ¹ H NMR (500 MHz, CDCl ₃ ) δ 7.72 (1H, d, J = 2.5 Hz), 7.36 (4H, s), 7.30 (1H, t, J = 8.5 Hz), 7.05 (1H, d, J = 2.5 Hz) 2.5 Hz), 7.00 (1H, d, J = 8.5 Hz), 6.83 (1H, d, J = 8.0 Hz), 6.82 (1H, d, J = 8.0 Hz), 6.79 (1H, dd, J = 2.5, 8.0 Hz), 6.76 (1H, d, J = 2.5 Hz), 6.72 (1H, t, J = 2.5 Hz), 6.59 (1H, dd, J = 1.0, 17.0 Hz), 6.29 (1H, dd, J = 10.0, 17.0 Hz), 6.01 (1H, dd, J = 1.0, 10.0 Hz), 5.02 (2H, s), 4.72 (2H, br s), 4.03 (2H, s), 3.22 (2H, t, J = 5.5 Hz), 2.98 (2H, t, J = 5.5 Hz); ¹³ C NMR (500 MHz, CDCl ₃ ) δ 164.4, 159.8, 157.3, 151.7, 151.6, 145.0, 135.7, 135.2, 134.4, 133.9, 132.9, 130.2, 128.9, 128.8, 128.7, 127.9, 127.6, 126.9, 120.6, 115.6 , 114.8, 114.2, 113.2, 110.4, 69.4, 52.8, 48.7, 29.9; LCMS (ESI) m/z calcd for C ₃₀ H ₂₅ C ₁ N ₃ O ₄ , 527.16; Found 528.16 [M + H] ⁺ .

< Example 16> Acrylic acid 3-{6-amino-5-[6-(4- methoxybenzyloxy )-3,4- dihydro Preparation of -1H-isoquinolin-2-yl]pyridin-3-yloxy}phenyl ester

Acrylic acid 3-{ 6-amino-5-[6-(4-methoxybenzyloxy)-3,4-dihydro-1H-isoquinolin-2-yl]pyridin-3-yloxy}phenyl ester (Acrylic acid 3-{6 -amino-5-[6-(4-methoxybenzyloxy)-3,4-dihydro- 1H -isoquinolin-2-yl]pyridin-3-yloxy}phenyl ester) was prepared.

yellow solid (yield 42%); IR (neat, cm ^-1 ) 3345.48, 2980.40, 2970.51, 2930.34, 1740.92, 1609.63, 1513.79, 1462.81, 1400.63, 1379.59, 1304.07, 1241.82, 1153.82, 1025.10, 1001.88; ¹ H NMR (500 MHz, CDCl ₃ ) δ 7.70 (1H, d, J = 2.5 Hz), 7.33 (2H, d, J = 8.5 Hz), 7.28 (1H, t, J = 8.0 Hz), 7.03 (1H) , d, J = 2.5 Hz), 6.96 (1H, d, J = 8.5 Hz), 6.90 (2H, d, J = 8.5 Hz), 6.82-6.77 (3H, m), 6.76 (1H, d, J = 8.5 Hz) 2.0 Hz), 6.73 (1H, t, J = 2.5 Hz), 6.57 (1H, dd, J = 1.0, 17.5 Hz), 6.27 (1H, dd, J = 10.5, 17.5 Hz), 5.98 (1H, dd, J = 1.0, 10.5 Hz), 4.95 (2H, s), 4.82 (2H, br s), 4.00 (2H, s), 3.78 (3H, s), 3.18 (2H, t, J = 6.0 Hz), 2.95 (2H, t, J = 6.0 Hz); ¹³ C NMR (500 MHz, CDCl ₃ ) δ 164.4, 159.8, 159.6, 157.7, 151.7, 151.7, 145.0, 135.3, 135.1, 134.4, 132.9, 130.2, 129.3, 129.2, 127.9, 127.5, 126.8, 120.6, 115.6, 114.8 , 114.2, 114.2, 113.3, 110.4, 70.0, 55.5, 52.8, 48.8, 29.9; LCMS (ESI) m/z calcd for C ₃₁ H ₂₉ N ₃ O ₅ , 523.21; Found 524.22 [M + H] ⁺ .

Chemical structures of the compounds prepared in Examples 1 to 16 are shown in Table 1 below.

No. chemical structure No. chemical structure One

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

< Experimental example 1> Molecular docking analysis

For molecular docking analysis of the compound according to Example 16, it was analyzed based on the human BTK structure (PDB ID: 3PJ3) with an allosteric pocket induced by DFG-out conformation. The results are shown in FIGS. 1 and 2 . As a result, it was confirmed that the tetrahydroisoquinoline linker of the compound of Example 16 generates a lipophilic moiety located in the allosteric pocket of the BTK kinase domain by the type II method. The aminopyridine hinge binder forms two hydrogen bonds with hinge Glu475 and gatekeeper Thr474, and the para-methoxybenzyl group is directed into an allosteric hydrophilic pocket surrounded by Met449, Leu452, Leu512 and Phe517. The lipophilic substituent is stabilized by a tetrahydroisoquinoline ring, which allows for van der Waals interactions with Lys430. In addition, a Michael acceptor linker phenyl ring exists in the gap between Leu408 and Gly480, through which a hydrogen bond is formed between the carbonyl of the acrylic group and the backbone of Cys481.

< Experimental example 2> BTK bow letter Inhibition Assessment

In order to evaluate the BTK inhibitory activity of Examples 1 to 16, BTK inhibitory activity and IC ₅₀ at a single concentration (20 μM) were measured by ADP-Glo assay (Promega, USA) and HotSpot Kinase assay (Reaction Biology Corporation, USA) was analyzed at an ATP concentration of 10 μM. The results are shown in Table 2 below.

No. BTK inhibitory activity (%) IC ₅₀ (μM) One 14 - 2 85 3.17 3 83 - 4 92 1.54 5 89 5.01 6 52 - 7 77 - 8 98 0.63 9 98 0.46 10 98 2.20 11 99 0.97 12 96 1.20 13 97 0.10 14 53 8.53 15 98 1.00 16 92 0.19

As can be seen in Table 2, the aminopyridine-based compound according to the present invention was found to have excellent activity in inhibiting BTK.

< Experimental example 3> BTK Evaluation of specificity and selectivity

In order to analyze the selectivity to BTK and the selectivity to other TEC and SRC family kinases, Example 9, Example 13, and Example 16 were tested in the same manner as in Experimental Example 2 using the compounds. The results are shown in Table 3 below.

kinase BTK inhibitory activity (%) Example 9 Example 13 Example 16 BMX/ETK 98.7 95.5 96.0 EGFR 87.0 60.5 33.5 ERBB2 43.4 82.3 78.8 ERBB4 95.0 96.2 95.0 ITK 75.1 52.8 46.0 JAK3 64.8 46.2 77.1 TEC 84.1 71.3 66.4 TXK 88.8 37.5 49.7

As shown in Table 3, the compound of the present invention was confirmed to have high specificity and selectivity to BTK. From the above results, the compound of the present invention has a scaffold different from that of the existing BTK inhibitor compound, and is appropriately located at the BTK active site to exhibit excellent binding force, and has high specificity and selectivity for BTK, thereby reducing side effects. It has been confirmed that a drug that lowers concerns and achieves more improved efficacy in the prevention or treatment of BTK-related diseases can be provided.

< Experimental example 4> Effect analysis on hematological malignancy

4-1. In vitro growth inhibition experiment

In order to analyze the effect of the compound of the present invention on hematological malignancy, first, the cell viability of Example 9 and Example 16 was analyzed for cytotoxicity and CellTiter-Glo luminescent cell viability. Raji (ATCC CCL-86) and Ramos (ATCC CRL-1596) cells were inoculated into opaque-walled 96-well plates (1 × 104 cells/well) and treated with various concentrations of Example 9 and Example 16 compounds. . After 48 hours, 100 μL of CellTiter-Glo Reagent (Promega, USA) was added to each well and the plate was incubated for 10 minutes at room temperature. The luminescence signal was measured using the GloMax-Multi Detection System (Promega, USA), and the GI ₅₀ value was calculated using GraphPad Prism (GraphPad, USA). The results are shown in Table 4 below.

GI ₅₀ (μM) Raji cell Ramos cell Example 9 16.1 12.5 Example 16 25.4 16.2

As a result, both compounds showed similar GI ₅₀ values in Raji and Ramos cell lines.

4-2. In in vivo xenotransplantation ( xenograft ) model analysis

In order to evaluate the in vivo activity of the compounds of Examples 9 and 16 of the present invention, an experiment was performed as follows. Specifically, the experiment was conducted with IACUC (Institutional Animal Care and Use Committee) approval of the Korea Research Institute of Bioscience and Biotechnology, and 6-week-old female BALB/c nu/nu mice (Orient Bio, Korea) Raji cells (9x10 ⁶ cells/head) were subcutaneously inoculated into the right flank of After 1 week, mice were randomized and administered vehicle (5% DMSO and 5% Tween 80 in corn oil), 2 mg/kg doxorubicin hydrochloride, 50 mg/kg Example 9 and Example 16 compounds. Treated 5 times a week. The tumor volume was measured with vernier calipers, and the length (mm) × width (mm) × height (mm)/2 was estimated by the formula, and the body weight was monitored three times a week. On day 14, mice were sacrificed and tumor weights were measured. Statistical analysis was performed using GraphPad Prism (GraphPad, USA). Changes in body weight and tumor volume were analyzed using two-way ANOVA and Bonferroni multiple comparison test, and p values of <0.05 were considered statistically significant. The results are shown in FIGS. 3 and 4 .

As a result, there were no toxic symptoms based on clinical and autopsy results for both compounds. The control group showed a slight weight loss, whereas the Example 9 and Example 16 compound groups showed no weight loss. In addition, Example 16 was shown to significantly reduce tumor size with an inhibitory effect of 46.8% compared to vehicle. Therefore, it can be seen that the compound of the present invention has an excellent effect in the treatment and improvement of hematologic malignancies.

As mentioned above, although the present invention has been described in detail through preferred preparation examples, examples and experimental examples, the scope of the present invention is not limited to the characteristic examples, and should be interpreted by the appended claims. In addition, those skilled in the art will understand that many modifications and variations are possible without departing from the scope of the present invention.

Claims (13)

A compound represented by the following formula (1), a stereoisomer or a pharmaceutically acceptable salt thereof:
[Formula 1]

In Formula 1,
wherein X ₁ is

or

ego;
wherein R ₂ is hydrogen or —OR ₃ ;
Wherein R ₁ and R ₃ are each independently hydrogen, a substituted or unsubstituted straight-chain or branched C _{1-10 alkyl} , or a substituted or unsubstituted C _1-10 alkoxycarbonyl,
wherein said substituted straight _- chain or branched C _{1-10 alkyl} , and C _1-10 alkoxy are each halogen, phenyl unsubstituted or substituted with one or more C _1-5 alkoxy or halogen, _and C _{3-8 cycloalkyl} substituted with one or more substituents selected from the group consisting of; and
wherein X ₂ is

am.
According to claim 1,
Wherein R ₁ and R ₃ are each independently hydrogen, a substituted or unsubstituted straight-chain or branched C _1-6 alkyl, or a substituted or unsubstituted C _1-6 alkoxycarbonyl,
wherein said substituted straight _{- chain} or branched C _{1-6 alkyl} , and C _1-6 alkoxy are each halogen, phenyl unsubstituted or substituted with one or more C _1-3 alkoxy or halogen, and C _{3-6 cycloalkyl} A compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, characterized in that it is substituted with one or more substituents selected from the group consisting of.
According to claim 1,
Wherein X ₂ Is ortho (ortho) or meta (meta), characterized in that substituted at the site, a compound, a stereoisomer or a pharmaceutically acceptable salt thereof.
According to claim 1,
The R ₁ and R ₃ are each independently hydrogen,

,

,

,

,

,

,

,

, or

Characterized in that, a compound, a stereoisomer or a pharmaceutically acceptable salt thereof.
According to claim 1,
The compound represented by Formula 1 is,
A compound, characterized in that it is any one selected from the group of compounds, a stereoisomer or a pharmaceutically acceptable salt thereof:
(1) acrylic acid 3-(6-amino-5-piperazinylpyridin-3-yloxy)phenyl ester;
(2) acrylic acid 3-[6-amino-5-(4-benzylpiperazinyl)pyridin-3-yloxy]phenyl ester;
(3) acrylic acid 3-{6-amino-5-[4-(4-fluorobenzyl)piperazinyl]pyridin-3-yloxy}phenyl ester;
(4) acrylic acid 3-{6-amino-5-[4-(4-chlorobenzyl)piperazinyl]pyridin-3-yloxy}phenyl ester;
(5) acrylic acid 3-[6-amino-5-(4-phenethylpiperazinyl)pyridin-3-yloxy]phenyl ester;
(6) acrylic acid 3-(6-amino-5-{4-[2-(4-chlorophenyl)ethyl]piperazinyl}pyridin-3-yloxy)phenyl ester;
(7) 4-[5-(3-acryloyloxyphenoxy)-2-aminopyridin-3-yl]piperazine carboxylic acid benzylester;
(8) acrylic acid 2-[6-amino-5-(3,4-dihydro-1H-isoquinolin-2-yl)pyridin-3-yloxy]phenyl ester;
(9) acrylic acid 2-[6-amino-5-(6-benzyloxy-3,4-dihydro-1H-isoquinolin-2-yl)pyridin-3-yloxy]phenyl ester;
(10) acrylic acid 3-[6-amino-5-(3,4-dihydro-1H-isoquinolin-2-yl)pyridin-3-yloxy]phenylester;
(11) acrylic acid 3-[6-amino-5-(6-propooxy-3,4-dihydro-1H-isoquinolin-2-yl)pyridin-3-yloxyphenyl ester;
(12) acrylic acid 3-[6-amino-5-(6-cyclopropylmethoxy-3,4-dihydro-1H-isoquinolin-2-yl)pyridin-3-yloxy]phenyl ester;
(13) acrylic acid 3-[6-amino-5-(6-benzyloxy-3,4-dihydro-1H-isoquinolin-2-yl)pyridin-3-yloxy]phenylester;
(14) acrylic acid 3-{6-amino-5-[6-(4-fluorobenzyloxy)-3,4-dihydro-1H-isoquinolin-2-yl]pyridin-3-yloxy}phenyl ester ;
(15) acrylic acid 3-{6-amino-5-[6-(4-chlorobenzyloxy)-3,4-dihydro-1H-isoquinolin-2-yl]pyridin-3-yloxy}phenyl ester; and
(16) Acrylic acid 3-{6-amino-5-[6-(4-methoxybenzyloxy)-3,4-dihydro-1H-isoquinolin-2-yl]pyridin-3-yloxy}phenyl ester .
A pharmaceutical composition for preventing or treating a BTK (Bruton's tyrosine kinase)-related disease comprising a compound represented by the following formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient:
[Formula 1]

In Formula 1,
X ₁ and X ₂ are as defined in Formula 1 of claim 1.
7. The method of claim 6,
The BTK-related disease is an autoimmune disease or a pharmaceutical composition, characterized in that cancer.
7. The method of claim 6,
The BTK-related disease is hematological malignancy, hematologic cancer, B-cell chronic lymphocytic leukemia, acute lymphoblastic leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma, acute myeloid leukemia, diffuse giant B-cell lymphoma, multiple A pharmaceutical composition selected from the group consisting of myeloma, jacket cell lymphoma, small lymphocytic lymphoma, mantle cell lymphoma (MCL) and lymphocytic leukemia (CLL).
7. The method of claim 6,
The BTK-related disease is selected from the group consisting of rheumatoid arthritis, osteoarthritis, juvenile arthritis, chronic obstructive pulmonary disease, multiple sclerosis, asthma, systemic lupus erythematosus, psoriasis, psoriatic arthritis, Crohn's disease, ulcerative colitis and irritable bowel syndrome. Characterized in that, the pharmaceutical composition.
7. The method of claim 6,
The BTK-related disease is characterized in that the hematological malignancy (hematological malignancy), the pharmaceutical composition.
A health functional food composition for preventing or improving BTK (Bruton's tyrosine kinase)-related disease containing a compound represented by the following formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient:
[Formula 1]

In Formula 1,
X ₁ and X ₂ are as defined in Formula 1 of claim 1.
12. The method of claim 11,
The BTK-related disease is an autoimmune disease or a health functional food composition, characterized in that cancer.
12. The method of claim 11,
The BTK-related disease is characterized in that the hematological malignancy (hematological malignancy), health functional food composition.

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