KR100885692B1 - 2-3-chloro-4-methoxyanilino-4-arylthiazole derivatives or pharmaceutically acceptable salts thereof, preparation method thereof, and phrmaceutical composition for the prevention and treatment of cancers containing the same as an active ingredient - Google Patents

Abstract

The present invention provides a 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative or a pharmaceutically acceptable salt thereof, a preparation method thereof, and a pharmaceutical for preventing and treating cancer containing the same as an active ingredient. In particular, the 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative represented by the following general formula (1) of the present invention is a vascular endothelial growth factor (VEGF) or epidermal growth factor By exhibiting the inhibitory effect of (EGF) receptor on tyrosine kinase, and the cytotoxic effect on cell growth factor overexpressing cancer cell lines, the pharmaceutical composition of the present invention inhibits neovascularization and cancer cell growth signaling, resulting in tumor cells It can be usefully used as an anticancer agent that inhibits the growth and metastasis of.

[Formula 1]

(Wherein R ¹ and R ² are as defined in the specification).

2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative, anticancer agent

Description

2- (3-Chloro-4-methoxy) anilino-4-arylthiazole derivative or pharmaceutically acceptable salt thereof, preparation method thereof and pharmaceutical composition for preventing and treating cancer containing same as an active ingredient {2 -(3-chloro-4-methoxy) anilino-4-arylthiazole derivatives or 黄 acceptable salts according to the method of preparation, especially, and phrmaceutical composition for the prevention and treatment of cancers containing the same as an active ingredient}

The present invention provides a 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative or a pharmaceutically acceptable salt thereof, a preparation method thereof, and a pharmaceutical for preventing and treating cancer containing the same as an active ingredient. It relates to a composition.

Receptor Tyrosine Kinases (RTKs) are cell membrane proteins that are a type of cell growth receptor and have the activity of tyrosine kinases regulated by endogenous ligands (Gschwind, A. et al., Nature review cancer, 4: 361). -370 (2004)). Outside the cell is the site where the ligand binds, and inside the cell is the tyrosine kinase site. RTKs regulate a variety of physiological functions, mainly through the signaling involved in somatic cell division of eukaryotic cells (Ullrich A. et al., Cell, 61: 623-634 (1990)). When certain cell growth factors, which are endogenous ligands, bind outside the RTKs cells, RTKs form cell duplexes by dimerization and autophosphorylation. The activity of RTKs is very tightly regulated in normal cells, but abnormal activation excessively activates cell signaling, leading to uncontrolled cell division and proliferation. Abnormal activation by gene mutation, amplification, and overexpression of RTKs is known to be involved in the development and progression of various human tumors and play a decisive role in the growth and metastasis of cancer cells.

Tumors are caused by mutations in the gene (DNA) and progress to other areas due to uncontrolled cell growth and cancer cell migration. Conventional chemotherapeutic agents act to kill cancer cells by inhibiting cell division through inhibition of DNA function and synthesis, but not by tumor cells but also by non-malignant normal cells by inhibiting cell division by preventing gastrointestinal disorders, hair loss, immunosuppression and Causes the same serious side effects. Toxicity and side effects due to the low selectivity between cancer cells and normal cells have been recognized as a problem to be continuously solved in the development of anticancer drugs. Recently, it has been shown to increase efficacy by selectively killing tumor cells or inhibiting growth without affecting normal cells. Tumor cell target anticancer drugs are expected to reduce side effects. In particular, studies have been actively conducted on substances displaying anticancer effects through inhibition of receptor tyrosine kinase (RTK), which regulates the signaling of cancer cell growth factor receptors (Bridges AJ, Chemical Reviews, 101: 2541-2572 (2001). )).

The growth of cancer cells is caused by uncontrolled division caused by genetic mutation, that is, cells do not respond to external signals that inhibit growth or overreact to growth promoting signals. Since RTKs are mainly involved in the signaling pathways for external and internal responses to cell growth, inhibition of RTKs can inhibit tumor cell growth and even cause tumor cell death. In addition, the cells can only survive within 100 ~ 200 ㎛ of the blood vessels, without the supply of the tumor can not grow more than a certain size. In order for the tumor to progress, tumor vascularization must occur by an angiogenic switch in which the balance between angiogenesis inducers and inhibitors changes, and tumors grow rapidly. Angiogenesis is also essential for tumor metastasis, and it is known that there is a direct correlation between tumor microvascular concentration and the survival rate of various solid cancers.

Changes in various angiogenesis-related factors are involved in tumors, but overexpression of VEGF, a type of RTK, is most clearly found. Induction of VEGF overexpression by tumors results in structural and functional non-vascular angiogenesis, leading to intratumoral hypoxia, which accelerates neovascularization (Ferrara, N., Nature review cancer, 2: 795-803 (2002)). Therefore, inhibition of RTKs can inhibit tumor growth and metastasis by inhibiting cancer cell or neovascularization. RTKs are mainly classified according to the type of endogenous ligand, EGFR (epidermal growth factor receptor), HER-2, vascular endothelial growth factor receptor type 2 (VEGFR-2), platelet-derived growth factor receptor (PDGFR), and IGFR (insulin). Research into the development of low molecular weight organic compounds targeting monoculon antibodies and tyrosine kinases has been actively conducted.

Hernthin of Genentech (1998) HER-2 antibody (Fendlly, BM et. Al., Cancer Research, 50; 1550-1558 (1990)), Novartis, 2001 Bcr-Abl and PDGFR tyrosine kinase inhibitors The low molecular organic compound Gleevec-Druka, BJ et. Al., Nature Medicine, 2: 561-566 (1996) has been FDA approved, demonstrating that RTKs are effective as targets for new tumor therapies. Subsequently, monoclonal antibodies include Avastin of Genentech, a VEGFR-2 antibody (2004-Presta, LG et. Al., Cancer Research, 57: 4593-4599 (1997)), and Imclone, an EGFR antibody. Erbitux (2004) was released. Low molecular weight compounds include Astra-Geneca's Iressa (Iressa, EGFR, 2002- Barker, AJ et. Al., Bioorganic & Medicinal Chemistry Letters, 11: 1911-1914 (2001)), Genentech / Roche Tarceva (EGFR, 2004) and Paiza's Sutant (KDR, 2006) have been approved for release of RTK inhibitors, and many other compounds are under clinical development.

RTKs inhibitors have been shown to be effective in treating tumors with poor treatment rates and prognosis with conventional chemotherapeutic anticancer agents such as metastatic breast cancer, gastrointestinal stromal tumor (GIST) and non-small cell lung cancer (NSCLC) in human cancer cell transplantation animal models and clinical studies. It is becoming.

The inventors of the present invention, while studying the inhibitors of VEGFR-2 and EGFR tyrosine kinase, synthesized 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivatives, and these compounds are receptor tyrosine kinase. By showing an inhibitory effect on, it was confirmed that there is a growth inhibitory effect of tumor cells, and completed the present invention.

It is an object of the present invention to provide a 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative or a pharmaceutically acceptable salt thereof.

 Another object of the present invention is to provide a method for preparing 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative or a pharmaceutically acceptable salt thereof.

Still another object of the present invention is a pharmaceutical composition for preventing and treating cancer, containing 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative or a pharmaceutically acceptable salt thereof as an active ingredient. To provide.

In order to achieve the above object, the present invention is a 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative or a pharmaceutically acceptable salt thereof, a method for preparing the same and containing the same as an active ingredient It provides a pharmaceutical composition for preventing and treating cancer.

Hereinafter, the present invention will be described in detail.

The present invention provides a 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

Where

R ¹ and R ² are independently or optionally hydrogen, hydroxy, halogen, nitro, cyano, C ₁ to C ₅ straight or branched chain alkyl, unsubstituted or halogen substituted, unsubstituted or substituted with halogen. 5-membered or 6-membered aryl, 5- or 6-membered heteroaryl containing N, 5- or 6-membered heterocyclic compound containing N, carbonyl substituted with amine, -NR ³ R ⁴ , -NHCOR ⁵ or is -OR ^6, wherein, R ³ and R ⁴ are independently or optionally, hydrogen, C ₁ ~ is a straight or branched chain alkyl or methanesulfonyl of C _5, R ⁵ is an unsubstituted or substituted by halogen C ₁ ~ C ₅ is a linear or branched alkyl, or an acrylic, R ⁶ is unsubstituted or N, and O at least one of C ₁ ~ C ₅ substituted with a heterocyclic compound of the 5-to 6-membered straight or branched chain containing from alkyl, Selected from the group consisting of trifluoromethyl and methanesulfonyl 1 which is a species.

Preferably,

,

,

, -CONH₂, -NR³R⁴, -NHCOR⁵ 또는 -OR⁶이고, 이때, R³ 및 R⁴는 선택적으로 또는 독립적으로, 수소, 메틸, 부틸 또는 메탄설포닐이고, R⁵는 -CH₂CH₂Br 또는 아크릴이며, R⁶는 메틸, 트리플루오로메틸,

,

,

또는

이다.R ¹ and R ² are optionally or independently hydrogen, hydroxy, F, Cl, Br, I, nitro, cyano, methyl, trifluoromethyl, chlorophenyl, pyridinyl,

,

,

, -CONH ₂ , -NR ³ R ⁴ , -NHCOR ⁵ or -OR ⁶ , wherein R ³ and R ⁴ are optionally or independently, hydrogen, methyl, butyl or methanesulfonyl, and R ⁵ is -CH ₂ CH ₂ Br or acryl, R ⁶ is methyl, trifluoromethyl,

,

,

or

to be.

Preferred examples of the 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative of the novel structure represented by Chemical Formula 1 according to the present invention are as follows.

1) 2- (3-chloro-4-methoxyanilino) -4-phenylthiazole;

2) 2- (3-chloro-4-methoxyanilino) -4- (3-bromophenyl) thiazole;

3) 2- (3-chloro-4-methoxyanilino) -4- (3-carbamoylphenyl) thiazole;

4) 2- (3-chloro-4-methoxyanilino) -4- (3-nitrophenyl) thiazole;

5) 2- (3-chloro-4-methoxyanilino) -4- (3-aminophenyl) thiazole;

6) 2- (3-chloro-4-methoxyanilino) -4- (3- (pyrrolidin-2-yl) phenyl) thiazole;

7) 2- (3-chloro-4-methoxyanilino) -4- (3- (pyrrole-1-yl) phenyl) thiazole;

8) 2- (3-chloro-4-methoxyanilino) -5- (3- (piperidin-yl) phenyl) thiazole;

9) 2- (3-chloro-4-methoxyanilino) -4- (3- (pyridin-2-yl) phenyl) thiazole;

10) 2- (3-chloro-4-methoxyanilino) -4- (3- (3-bromopropionamido) phenyl) thiazole;

11) 2- (3-chloro-4-methoxyanilino) -4- (3-acrylamidophenyl) thiazole;

12) 2- (3-chloro-4-methoxyanilino) -4- (3-methanesulfonamidophenyl) thiazole;

13) 2- (3-chloro-4-methoxyanilino) -4- (3-dimethanesulfonamidophenyl) thiazole;

14) 2- (3-chloro-4-methoxyanilino) -4- (3-hydroxyphenyl) thiazole;

15) 2- (3-chloro-4-methoxyanilino) -4- (3- (2- (pyrrolidin-1-yl) ethoxy) phenyl) thiazole;

16) 2- (3-chloro-4-methoxyanilino) -4- (3- (2- (piperidin-1-yl) ethoxy) phenyl) thiazole;

17) 2- (3-chloro-4-methoxyanilino) -4- (3- (2- (morpholin-1-yl) ethoxy) phenyl) thiazole;

18) 2- (3-chloro-4-methoxyanilino) -4- (4-chlorophenyl) thiazole;

19) 2- (3-chloro-4-methoxyanilino) -4- (4-fluorophenyl) thiazole;

20) 2- (3-chloro-4-methoxyanilino) -4- (4-cyanophenyl) thiazole;

21) 2- (3-chloro-4-methoxyanilino) -4- (4-bromophenyl) thiazole;

22) 2- (3-chloro-4-methoxyanilino) -4- (4-methylphenyl) thiazole;

23) 2- (3-chloro-4-methoxyanilino) -4- (4-trifluoromethylphenyl) thiazole;

24) 2- (3-chloro-4-methoxyanilino) -4- (4-iodophenyl) thiazole;

25) 2- (3-chloro-4-methoxyanilino) -4- (4-carbamoylphenyl) thiazole;

26) 2- (3-chloro-4-methoxyanilino) -4- (4-nitrophenyl) thiazole;

27) 2- (3-chloro-4-methoxyanilino) -4- (4-aminophenyl) thiazole;

28) 2- (3-chloro-4-methoxyanilino) -4- (4-N-methylaminophenyl) thiazole;

29) 2- (3-chloro-4-methoxyanilino) -4- (4-N; N-dimethylaminophenyl) thiazole;

30) 2- (3-chloro-4-methoxyanilino) -4- (4-N-1-butylaminophenyl) thiazole;

31) 2- (3-chloro-4-methoxyanilino) -4- (4- (pyrrolidin-1-yl) phenyl) thiazole;

32) 2- (3-chloro-4-methoxyanilino) -4- (4- (piperidin-1-yl) phenyl) thiazole;

33) 2- (3-chloro-4-methoxyanilino) -4- (4- (pyrrole-1-yl) phenyl) thiazole;

34) 2- (3-chloro-4-methoxyanilino) -4- (4- (3-bromopropionamido) phenyl) thiazole;

35) 2- (3-chloro-4-methoxyanilino) -4- (4-acrylamidophenyl) thiazole;

36) 2- (3-chloro-4-methoxyanilino) -4- (4-hydroxyphenyl) thiazole;

37) 2- (3-chloro-4-methoxyanilino) -4- (4- (2- (piperidin-1-yl) ethoxy) phenyl) thiazole;

38) 2- (3-chloro-4-methoxyanilino) -4- (4- (2- (pyrrolidin-1-yl) ethoxy) phenyl) thiazole;

39) 2- (3-chloro-4-methoxyanilino) -4- (4- (2- (morpholin-1-yl) ethoxy) phenyl) thiazole;

40) 2- (3-chloro-4-methoxyanilino) -4- (4- (3- (piperidin-1-yl) propoxy) phenyl) thiazole;

41) 2- (3-chloro-4-methoxyanilino) -4- (2; 4-dichlorophenyl) thiazole;

42) 2- (3-chloro-4-methoxyanilino) -4- (4-methoxy-3-nitrophenyl) thiazole;

43) 2- (3-chloro-4-methoxyanilino) -4- (3-amino-4-methoxyphenyl) thiazole;

44) 2- (3-chloro-4-methoxyanilino) -4- (4-bromo-3-nitrophenyl) thiazole;

45) 2- (3-chloro-4-methoxyanilino) -4- (3-amino-4-bromophenyl) thiazole;

46) 2- (3-chloro-4-methoxyanilino) -4- (4-bromo-3- (piperidin-1-yl) phenyl) thiazole;

47) 2- (3-chloro-4-methoxyanilino) -4- (4-bromo-3-pyrrolylphenyl) thiazole;

48) 2- (3-chloro-4-methoxyanilino) -4- (4-chlorobisphenyl-4-yl) thiazole;

49) 2- (3-chloro-4-methoxyanilino) -4- (4-chloro-3-nitrophenyl) thiazole;

50) 2- (3-chloro-4-methoxyanilino) -4- (3-amino-4-chlorophenyl) thiazole;

51) 2- (3-chloro-4-methoxyanilino) -4- (4-chloro-3- (piperidin-1-yl) phenyl) thiazole; or

52) 2- (3-chloro-4-methoxyanilino) -4-((4-chloro-3-pyrrole-1 yl) phenyl)) thiazole.

2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative represented by Chemical Formula 1 according to the present invention may be used in the form of a pharmaceutically acceptable salt. As the salts, acid addition salts formed by various organic or inorganic acids that are pharmaceutically or physiologically acceptable are useful. Suitable organic acids include, for example, carboxylic acid, phosphonic acid, sulfonic acid, acetic acid, propionic acid, octanoic acid, decanoic acid, glycolic acid, lactic acid, fumaric acid, succinic acid, adipic acid, malic acid, tartaric acid, citric acid, glutamic acid, aspartic acid, Maleic acid, benzoic acid, salicylic acid, phthalic acid, phenylacetic acid, benzenesulfonic acid, 2-naphthalenesulfonic acid, methyl sulfuric acid, ethyl sulfuric acid, dodecyl sulfuric acid, and the like can be used. Examples of suitable inorganic acids include hydrochloric acid, sulfuric acid, phosphoric acid, and the like. Can be used.

In addition, 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative represented by Chemical Formula 1 according to the present invention may be prepared by conventional methods as well as pharmaceutically acceptable salts. All possible salts, hydrates, and solvates may be included.

The present invention also provides a method for preparing 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof.

Specifically, the preparation method of the 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative according to the present invention, as shown in Scheme 1, a metal catalyst of the compound of formula 2 as a starting material And condensation with 3-chloro-p-acinidine of Formula 3 in the presence of an organic solvent to obtain a compound of Formula 1 (Step 1).

(In the above formula, R ¹ and R ² are the same as in the general formula (1).)

In the production method of the present invention, after reacting the α-bromoacetophenone compound of the formula (2) with a thiocyanate salt (MSCN) of an alkali metal such as sodium, ammonium, potassium, lithium, and the like, the equivalent of 3- Compound (1) can be prepared by one step reaction by condensation reaction with chloro-p-anisidine (3). At this time, the organic solvent may be an alcohol solvent such as methanol, ethanol and isopropanol, ether solvent such as dioxane and dimethylformamide (DMF). The reaction temperature is preferably a temperature from normal temperature to the boiling point of the solvent.

The present invention also provides another method for preparing 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof.

Specifically, another method for preparing 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative according to the present invention, as shown in Scheme 2 below, the starting material of the compound of formula (2) And condensation reaction with N- (3-chloro-4-methoxyphenyl) thiourea of formula 4 in the presence of an organic solvent to obtain a compound of formula 1 (step 1).

(In the above formula, R ¹ and R ² are the same as in the general formula (1).)

In another production method of the present invention, the 2-bromoacetophenone compound of the formula (2) is reacted with 2- by condensation reaction with N- (3-chloro-4-methoxyphenyl) thiourea (4) (3-Chloro-4-methoxy) anilino-4-arylthiazole derivative (1) can be prepared. In this case, alcohol solvents such as methanol, ethanol and isopropanol, ether solvents such as dioxane, and dimethylformamide (DMF) may be used. The reaction temperature is preferably a temperature from normal temperature to the boiling point of the solvent.

In addition, the production method of the present invention may further include a step of substituting with another substituent.

,

,

, -NR³R⁴인 아미노 유도체(1a)는 R¹이 -NO₂인 화합물을 사용하여 제조할 수 있다.1. As shown in Scheme 3 below, R ¹ of the 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative of the present invention is -NH ₂ ,

,

,

The amino derivative (1a) having -NR ³ R ⁴ can be prepared using a compound wherein R ¹ is -NO ₂ .

(Wherein R ¹ , R ² , R ³ and R ⁴ are the same as in Formula 1, Hal is a halogen element, and m is 1 or 2.)

1) Compound (1a ₂ ) wherein R ¹ is -NH ₂ can be obtained by reduction of compound (1a ₁ ) wherein R ¹ is -NO ₂ . In this case, the -NO ₂ group may be reduced to -NH ₂ by hydrogenation in the presence of an organic solvent and a metal catalyst such as platinum, palladium-attached charcoal (Pd / C; palladium on carbon) or Raney-nickel. The organic solvent may be an alcohol solvent such as methanol, ethanol or ethyl acetate. Alternatively, it may be reduced with a reducing agent such as sodium borohydride (NaBH ₄ ) in the presence of a catalyst such as CuSO ₄ , Cu (OAc) ₂ , CoCl ₂ or NiCl ₂ . At this time, it is preferable to use the mixed solvent of water and methanol as a reaction solvent, and it is preferable to make reaction temperature into normal temperature.

2) R ¹ is -NHR ³ of compound (1a _3), the form is when R ¹ is -NH ₂ in the compound of (1a ₂₎ a C ₁ ~ C ₅ aldehyde (R ³ of the reducing agent and an organic solvent under the presence methyl It can be obtained by reductive alkylation using aqueous aldehyde solution or paraformaldehyde (solid). In this case, as the reducing agent, sodium borohydride (NaBH ₄ ), sodium cyanoborohydride (NaBH ₃ CN), and the like may be used, and as an organic solvent, an alcohol solvent such as methanol, ethanol, tetrahydrofuran, or the like may be used. Ether solvents, ethyl acetate and the like can be used, the reaction temperature is preferably the boiling point temperature of the solvent at zero.

3) A dialkylamino compound (1a ₄ ) wherein R ¹ is —NR ³ R ⁴ is a dialkylation of a compound (1a ₂ ) wherein R ¹ is —NH ₂ with a halogenated alkyl in the presence of a base and an organic solvent, or R ¹ is- alkylating an amino group NR ³ H of the compound (1a ₃₎ to be obtained. In this case, the halogen may be bromine, chlorine or iodine. Examples of the base used for the dialkylation or alkylation reaction include inorganic bases such as sodium hydride, potassium t-butoxide and sodium methoxide, or N, N-diisopropylamine, 1,8-diazabicyclo [ 5,4,0] undec-7-ene (DBU) and the like can be used. The organic solvent may be used alone or mixed with an ether solvent such as tetrahydrofuran, dioxane, 1,2-dimethoxyethane, DMF, dimethyl sulfoxide (DMSO), or the like. Moreover, the temperature of reaction temperature from 0 to the boiling point of a solvent is preferable.

또는

인 고리알킬아미노 화합물(1a₅)은 R¹이 -NH₂인 화합물(1a₂)을 염기 및 유기 용매 존재 하에서 고리화 알킬화 반응시켜 얻을 수 있으며, 이때, 상기 염기, 유기 용매 및 반응 온도는 상기 디알킬화 또는 알킬화 반응 과 동일하게 수행될 수 있다.4) R ¹ is

or

Phosphorus cyclic alkylamino compound (1a ₅ ) can be obtained by cyclization alkylation of compound (1a ₂ ) wherein R ¹ is —NH ₂ in the presence of a base and an organic solvent, wherein the base, organic solvent and reaction temperature are It may be carried out in the same manner as the dealkylation or alkylation reaction.

인 피롤 화합물(1a₆)은 R¹이 -NH₂인 화합물(1a₂)을 고리화 반응시켜 얻을 수 있다. 2,5-디메톡시테트라히드로퓨란을 1 내지 1.5 당량 사용하여 아세트산 용매에서 가열 환류시키거나, 또는 4-클로로피리딘 히드로클로라이드 촉매를 사용하여 디옥산에서 가열 환류시켜 얻을 수 있다.5) R ¹ is

The inpyrrole compound (1a ₆ ) can be obtained by cyclizing a compound (1a ₂ ) in which R ¹ is -NH ₂ . It can be obtained by heating to reflux in an acetic acid solvent using 1 to 1.5 equivalents of 2,5-dimethoxytetrahydrofuran, or by heating to reflux in dioxane using a 4-chloropyridine hydrochloride catalyst.

2. As shown in Scheme 4 below, when R ¹ of the 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative of the present invention is -NHCOR ⁵ (1b), R ¹ is It can be obtained from -NH ₂ phosphorus compound (1a ₁ ) by using carboxylic acid chloride (R ⁵ COCl) in the presence of an organic solvent and a base.

(In the above formula, R ² and R ⁵ are the same as in Chemical Formula 1.)

In the production method of the present invention, the base may be a base such as triethylamine, N, N-diisopropylamine, pyridine, and the like, and the organic solvent is methylene chloride, chloroform, dimethylformamide, tetrahydrofuran Etc. can be used, and the reaction temperature is preferably 0 to room temperature.

(Wherein R ² is the same as that of Formula 1)

In this case, when R ¹ is acrylamide (1b ₂ ), as shown in Scheme 5, the compound (1a ₂ ) and 3-bromopropanoyl chloride in which R ¹ is -NH ₂ Substitution method of Scheme 4 Compound (1b ₁ ), which is 3-bromo propionamide obtained by reaction with, can be obtained by debromic acid reaction by heating to reflux in the presence of a base and an organic solvent. The base and the organic solvent are the same as the reaction for preparing the amide compound (1b).

3. As shown in Scheme 6 below, when R ¹ of the 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative of the present invention is -OR ⁶ (1c), R ¹ It can be obtained by reacting the compound (1a ₇ ) which is -OH in the presence of an organic solvent and a base.

(Wherein, R ² and R ⁶ are the same as in Formula 1, and Hal is halogen.)

When R ¹ is alkoxy, it can be obtained by alkylating the phenol compound (1c) with an alkyl halide in the presence of a base and an organic solvent. In this case, the halogen may be bromine, chlorine or iodine. The base may be an inorganic base such as sodium hydride, potassium t-butoxide, sodium methoxide, or an organic base such as N, N-diisopropylamine or DBU. As the organic solvent, ether solvents such as tetrahydrofuran, dioxane, 1,2-dimethoxyethane, DMF, DMSO, and the like may be used alone or in combination. In addition, the boiling temperature of the solvent is preferably 0 at the reaction temperature.

4. As shown in Scheme 7 below, when R ¹ of the 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative of the present invention is -CONH ₂ (1d), R ¹ The compound (1a ₈ ) which is -CN can be obtained by reacting in the presence of a reaction solvent and a base.

(Wherein R ² is the same as that of Formula 1)

When R ¹ is -CONH ₂ , the carbamoyl compound (1d) can be obtained by selective hydrolysis of the nitrile compound (1a ₈ ). At this time, it can be prepared by hydrolysis using inorganic bases such as potassium hydroxide or sodium hydroxide, and the use of hydrogen peroxide increases the selectivity of hydrolysis to carbamoyl. Alternatively, it can be prepared by hydrolyzing the nitrile compound with acid using acetic acid or trifluoroacetic acid and sulfuric acid. The reaction solvent may be water or an alcohol solvent such as methanol, ethanol, isopropanol, and the reaction temperature is preferably the boiling point temperature of the solvent at room temperature.

5. As shown in Scheme 8, when R ¹ of the 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative of the present invention is aryl (1e), R ¹ is halogen ( It can be obtained by reacting X) as a compound (1a ₉ ) in the presence of a metal catalyst, a base and a reaction solvent.

Wherein R ² is the same as Formula 1, Ar is aryl unsubstituted or substituted, X is halogen, Y is -B (OH) ₂ , -BCl ₂ , -BBr ₂ , -SnBu ₃ , -SnMe ₃ , or -ZnCl.)

When R ¹ is aryl (Ar), a halogen compound (1a ₉ ) and an arylboronic acid or stanylaryl derivative compound are obtained by a steel-type coupling or a Suzuki-type coupling reaction in the presence of a metal catalyst, especially a palladium catalyst. Can be.

In this case, the arylboronic acid or stanylyl compound may be used by commercially available compounds, or may be prepared by using a known method from aryl halides. Palladium, nickel, platinum complexes and the like may be used as the metal catalyst, but a palladium catalyst is preferably used. As the palladium catalyst, Pd (PPh ₃ ) ₄ , Pd-C, PdCl ₂ (PPh ₃ ) ₂ , Pd ₂ (dba) ₃ , PdCl ₂ (dppf), [PdCl (allyl)] ₂ , Pd (OAc) ₂ , PdCl ₂ and the like can be used. In addition, in order to promote the reaction and increase the yield, phosphine compounds such as PPh ₃ , P- (o-tolyl) ₃ , and PBu ₃ are used as adducts, or salts such as lithium chloride, lithium bromide, and lithium iodide are added. Can be used as water.

When the Suzuki-type reaction is performed in Scheme 8, 1 to 3 equivalents of the base may be used. At this time, examples of the base that can be used include tertiary amine organic bases such as triethylamine and isopropylethylamine, inorganic bases such as sodium carbonate, potassium carbonate, potassium hydroxide, sodium hydroxide, cesium carbonate and barium hydroxide. When the inorganic base is not dissolved in an organic solvent, the inorganic base may be added by dissolving it in water. In this case, the inorganic base may be used so that the concentration of the inorganic base is about 0.5 to 4 M. Examples of the reaction solvent include ether solvents such as tetrahydrofuran, dioxane and 1,2-dimethoxyethane, aromatic hydrocarbon solvents such as benzene, toluene and xylene, alcohol solvents such as methanol and ethanol, DMF and aceto. Nitrile, ethyl acetate and the like can be used alone or in combination. The temperature of the reaction is preferably room temperature to the boiling point temperature of the solvent.

Further, the present invention is to prevent cancer containing 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative represented by the formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient and Provided are a therapeutic pharmaceutical composition and a pharmaceutical composition for inhibiting angiogenesis.

In this case, the cancer is liver cancer, stomach cancer, breast cancer, colon cancer, bone cancer, pancreatic cancer, head or neck cancer, uterine cancer, ovarian cancer, rectal cancer, esophageal cancer, small intestine cancer, anal muscle cancer, colon cancer, fallopian tube carcinoma, endometrial carcinoma, cervical carcinoma , Vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, prostate cancer, bladder cancer, kidney cancer, ureter cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system tumor.

In the experiment conducted to confirm the RTK enzyme inhibitory effect of the 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative of the present invention (see Experimental Example 1), VEGFR-2 tyrosine kinase Tyrphostin A51, a control compound, showed a strong inhibitory effect of 88% at 20 μM, and the compounds of Examples 6, 8, 21, 28, 29, 37, 39-44, and 46 of the present invention were VEGFR- at 20 μM. 2 showed a good inhibitory effect of more than 60% against tyrosine kinase. In particular, the compounds of Examples 6, 21, 28 and 46 showed at least 70% inhibitory effect. The inhibitory effect on the VEGFR-2 tyrosine kinase is to inhibit the growth and metastasis of tumor cells through the inhibition of neovascularization.

In addition, Tyrphostin A51, a control against EGFR tyrosine kinase, showed a strong inhibitory effect of 93% at 20 μM and the compounds of Examples 6, 8, 21, 28, 40, 44, and 46 of the present invention had a concentration of 20 μM. Showed an inhibitory effect of more than 60% against EGFR tyrosine kinase. In particular, the compounds of Examples 6, 40 and 46 showed at least 70% inhibitory effect. The inhibitory effect on EGFR tyrosine kinase is to inhibit the growth and metastasis of tumor cells through cancer cell growth inhibition.

In addition, in an experiment conducted to determine in vitro cytotoxicity in order to measure the effect of inhibiting the growth of cancer cell lines, the 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivatives of the present invention ( Experimental Example 2), the compounds of Examples 16 and 17 exhibited an moderate inhibitory effect with an IC ₅₀ value of about 5 μM against SK-OV-3, a HER-2 overexpressing cell line, and A431, an EGF overexpressing cell line. In particular, the compound of Example 46 exhibited an IC ₅₀ value of 0.82 and 1.10 μM for HER-2 overexpressing cell lines SK-OV-3 and A431, respectively, indicating an excellent inhibitory effect on growth factor overexpressing cancer cell lines.

Accordingly, the 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivatives of the present invention inhibit the vascular endothelial growth factor (VEGF) or epithelial growth factor (EGF) receptors against tyrosine kinases. By exhibiting effects and showing cytotoxic effects on cell growth factor overexpressing cancer cell lines, the pharmaceutical compositions of the present invention inhibit neovascularization and cancer cell growth signaling, and thus are useful as anticancer agents that inhibit the growth and metastasis of tumor cells. Can be used.

The 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivatives or pharmaceutically acceptable salts thereof of the present invention can be administered in various dosage forms, oral and parenteral, during clinical administration. When formulated, it may be prepared using conventional diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, and the like.

Solid form preparations for oral administration include tablets, patients, powders, granules, capsules, troches and the like, and such solid form preparations include one or more of the 2- (3-chloro-4-methoxy) anilino-4 of the present invention. -Arylthiazole derivatives or pharmaceutically acceptable salts thereof can be prepared by mixing at least one excipient such as starch, calcium carbonate, sucrose or lactose or gelatin. In addition to the simple excipients, lubricants such as magnesium stearate, talc and the like can also be used. Liquid preparations for oral administration include suspensions, solvents, emulsions or syrups, and include various excipients such as wetting agents, sweeteners, fragrances, and preservatives in addition to the commonly used simple diluents, water and liquid paraffin. Can be.

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerol, gelatin and the like can be used.

In addition, the dosage of the 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative of the present invention or a pharmaceutically acceptable salt thereof to the human body is determined by the age, weight, sex, dosage form of the patient. , Depending on the state of health and the degree of disease, based on adult patients weighing 70 Kg, generally 0.1 to 1000 mg / day, preferably 1 to 500 mg / day, and also doctors or pharmacists Depending on the judgment of the administration may be divided once to several times a day at regular intervals.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are not limited to the contents of the present invention by the following examples.

1. Preparation of starting material.

The α-bromoacetophenone compound (2) used as a starting material in Scheme 1 or Scheme 2 may be a commercially available compound or known α- from acetophenone compound (5) as shown in Scheme 9 below. It can be manufactured and used by bromination reaction.

(In the above formula, R ¹ and R ² are the same as in the general formula (1).)

In the bromination reaction, Br ₂ , CuBr ₂ , N-bromosuccinimide (NBS), etc. may be used as the brominating agent, and as the reaction solvent, halogenated hydrocarbon solvents such as methylene chloride, chloroform, carbon tetrachloride, Ether solvents such as tetrahydrofuran, 1,4-dioxane or ethyl acetate, acetonitrile and acetic acid, bromic acid, sulfuric acid and the like can be used. The reaction temperature is preferably 0 to the boiling point temperature of the solvent.

In the starting material preparation reaction, the compound of formula (5) is subjected to the dibromination reaction at the α-position, and then the α-bromoacetophenone compound (2) may be obtained by the debromination reaction. The debromination reaction can be accomplished using diethylphosphite with a base. As the base, triethylamine, isopropylethylamine, and the like are used. As the reaction solvent, an ether solvent such as tetrahydrofuran and diethyl ether is preferably used. The reaction temperature is preferably 0 to a boiling point temperature of the solvent. Do.

The acetophenone compound (5) used as a starting material in Scheme 9 may be prepared by using a commercially available compound or by the following method.

,

및

인 아미노 유도체는 경우, R¹이 -NO₂인 화합물을 사용하여 상기 반응식 3의 치환반응과 동일한 방법으로 얻을 수 있다. 1) R ¹ of acetophenone compound (5) is -NH ₂ , -NHR ³ , -NR ³ R ⁴ ,

,

And

Phosphorus amino derivatives can be obtained in the same manner as the substitution reaction of Scheme 3 using a compound wherein R ¹ is -NO ₂ .

2) A compound in which R ¹ of the acetophenone compound (5) is -NHCOR ⁵ can be obtained by the same method as in Scheme 4 using a compound of R ¹ of -NH ₂ .

3) A compound in which R ¹ of the acetophenone compound (5) is -OR ⁶ can be obtained by the same method as in Scheme 6, using a compound in which R ¹ is -OH.

4) The compound in which R ¹ of the acetophenone compound (5) is -CONH ₂ can be obtained by the same method as in Scheme 7 using the compound of R ¹ of -CN.

5) The compound in which R ¹ of the acetophenone compound (5) is aryl can be obtained by the same method as in Scheme 7 using a compound in which R ¹ is halogen.

In addition, the acetophenone compound (5) used as a starting material in Scheme 9 may be prepared from the benzonitrile compound by another method, as shown in Scheme 10 below.

(Wherein R ¹ and R ² are the same as in Formula 1, and M is a metal salt.)

In Scheme 10, the acetophenone compound (5) can be obtained by adding methyl metal salt to the nitrile group and hydrolyzing it. As the metal salt, lithium, Grignard reagent (MgX, wherein X is Br or Cl), ZnCl, SmI, or the like may be used. It is preferable to use an ether solvent such as tetrahydrofuran, diethyl ether or the like as the reaction solvent, and the reaction temperature is preferably 0 to the boiling point temperature of the solvent. The hydrolysis is reacted in an aqueous solution such as hydrochloric acid or sulfuric acid, and the reaction temperature is preferably a boiling point temperature of room temperature to water.

N- (3-chloro-4-methoxyphenyl) thiourea (4) used as a starting material in Scheme 2 may be prepared and used as shown in Scheme 11 below.

As shown in Scheme 11 above, N- (3-chloro-4-methoxyphenyl) thiourea (4) is first used for benzoyl isothiocyanate (8) and 3-chloro-p-anisidine (3). By reaction, N- (3-chloro-4-methoxyphenyl) -N'-benzoylthiourea (9) should be prepared. In this case, acetone or acetonitrile may be used as the reaction solvent, and the reaction temperature is preferably from room temperature to the boiling point temperature of the solvent. Subsequently, the produced N- (3-chloro-4-methoxyphenyl) -N'-benzoylthiourea (9) was hydrolyzed using a base, thereby N- (3-chloro-4-methoxyphenyl) The thiourea (4) can be obtained. Examples of the base include inorganic bases such as sodium carbonate, potassium carbonate, potassium hydroxide, sodium hydroxide, cesium carbonate, barium hydroxide or hydrazine. As the reaction solvent, an alcohol solvent such as methanol, ethanol or isopropanol, or an ether solvent such as dioxane or tetrahydrofuran can be used. In addition, the reaction temperature is preferably from room temperature to the boiling point temperature of the solvent.

Preparation Example 1 Preparation of 3-carbamoyl acetophenone

3-acetylbenzonitrile (2.5 g, 17.22 mmol) was dissolved in ethanol (25 ml) and hydrogen peroxide (H ₂ O ₂ 27.5% in H ₂ O; 7.5 ml, 60.5 mmol) was added slowly at room temperature, followed by water (8.5 ml) was added and stirred at 50 ° C. for 1 hour. The ethanol was evaporated under reduced pressure, extracted with methylene chloride (CH ₂ Cl ₂ ; 80 ml) and washed with water and brine. The extract was dried over anhydrous magnesium sulfate (MgSO ₄ ), filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (hexane: ethyl acetate = 1: 2, v / v) to obtain a white solid target compound (2.74 g, 97%).

¹ H-NMR (200 MHz, CDCl ₃ ) σ 2.67 (s, 3H), 6.03 (bs, 1H), 6.20 (bs, 1H), 7.58 (t, 1H), 8.06 (dt, 1H), 8.12 (dt , 1H), 8.40 (s, 1 H);

Ms 163 (M ⁺ ).

Preparation Example 2 Preparation of 4-carbamoyl acetophenone

The reaction was carried out in the same manner as in Preparation Example 1 using 4-acetyl benzonitrile to obtain the title compound (2.72 g, 97%) as a yellow solid.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 2.61 (s, 3 H), 7.55 (s, 1 H), 8.02 (m, 4 H), 8.13 (s, 1 H);

Ms 163 (M ⁺ ).

Preparation Example 3 Preparation of 3- (pyridin-2-yl) acetophenone

3-bromoacetophenone (3 g, 15.07 mmol) was dissolved in toluene (30 ml) and Pd (PPh ₃ ) ₄ (867 mg, 5 mol%) and N-tributylstananyl-2-pyridine (N- tributylstannanyl-2-pyridine; 9 g, 18.08 mmol) was added at room temperature and heated to reflux for 12 hours. The reaction mixture was cooled to room temperature, extracted with ethyl acetate (150 ml) and washed with water and brine. The extract was dried over anhydrous MgSO ₄ , filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (hexane: ethyl acetate = 4: 1, v / v) to give an oily target compound (2.27 g, 55%). Got it.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 2.69 (s, 3H), 7.32 (m, 1H), 7.58 (dd, 1H), 7.80 (m, 2H), 8.00 (d, 1H), 8.23 (d , 1H), 8.58 (s, 1 H), 8.73 (m, 1 H);

Ms 197 (M ⁺ ).

Preparation Example 4 Preparation of 4- (4-chlorophenyl-4-yl) acetophenone

4-bromoacetophenone (230 mg, 1.15 mmol) was dissolved in toluene (5 ml) and Pd (PPh ₃ ) ₄ (55 mg, 5 mol%) and 4-chlorophenyl boronic acid (150 mg, 0.96 mmol) It was added at room temperature and then heated to reflux for 12 hours. Cooled to room temperature, extracted with ethyl acetate (150 ml) and washed with water and brine. The extract was dried over anhydrous MgSO ₄ , filtered and concentrated under reduced pressure, and purified by silica gel column chromatography (hexane: ethyl acetate = 8: 1, v / v) to give the title compound (170 mg, 77%) as a white solid. Got it.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 2.64 (s, 3H), 7.45 (dd, 2H), 7.54 (dd, 2H), 7.66 (d, 2H), 8.02 (d, 2H);

Ms 230 (M ⁺ ).

Preparation Example 5 Preparation of 4- (pyrrolidin-1-yl) acetophenone

4-aminoacetophenone (800 mg, 5.9 mmol) was dissolved in DMF (7 m) and NaH (448 mg, 17.7 mmol) was added at 0 ° C., followed by 1,4-dioodobutane (1.2 ml, 8.9 mmol). ) Was added. After stirring for 6 hours at room temperature, water (20 ml) was added, extracted with ethyl acetate (20 ml) and the organic layer was washed with water and brine. The extract was dried over anhydrous MgSO ₄ , filtered, concentrated under reduced pressure, and purified by silica gel chromatography (hexane: ethyl acetate = 4: 1, v / v) to give the title compound (330 mg, 30%) as a yellow solid. Got it.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 2.03 (m, 4H), 2.50 (s, 3H), 3.36 (m, 4H), 6.52 (d, 2H), 7.87 (d, 2H).

Preparation Example 6 Preparation of 4- (piperidin-1-yl) acetophenone

The reaction was carried out in the same manner as in Preparation Example 5 using 4-aminoacetophenone and 1,5-diazodobutane to obtain the title compound (323 mg, 27%) as a pale yellow solid.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 1.58 (m, 6H), 2.50 (s, 3H), 3.36 (m, 4H), 6.85 (d, 2H), 7.84 (d, 2H).

Preparation Example 7 Preparation of 3- (pyrrolidin-1-yl) acetophenone

Step 1. 3- ( Pyrrolidine -1 day) Benzonitrile  Produce

Using 3-aminobenzonitrile (1 g, 8.46 mmol) in the same manner as in Preparation Example 5 to obtain the target compound (1.02 g, 70%) in the form of an oil.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 2.04 (m, 4H), 3.27 (m, 4H), 6.74 (m, 2H), 6.80 (d, 1H), 7.25 (t, 1H).

Step 2. 3- ( Pyrrolidine -1-yl) Preparation of Acetophenone

3- (pyrrolidin-yl) benzonitrile (1.02 g, 5.90 mmol) obtained in step 1 was dissolved in anhydrous diethyl ether (8 ml) and methylmagnesium bromide (MeMgBr-3 M in diethyl ether; 3.93 ml , 11.8 mmol) was added and then heated to reflux for 12 hours. After cooling to room temperature, diethyl ether and ice water were added, followed by extraction with 1 N HCl. The aqueous layer was heated to reflux for 1 hour, followed by addition of aqueous sodium hydrogen carbonate (NaHCO ₃ ) solution, extraction with ethyl acetate (80 ml), and washing with water and brine. The extract was dried over anhydrous MgSO ₄ , filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (hexane: ethyl acetate = 9: 1, v / v) to give an oily target compound (991 mg, 89%). Got it.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 2.02 (m, 4H), 2.58 (s, 3H), 3.33 (m, 4H), 6.76 (dd, 1H), 7.13 (s, 1H), 7.23 (t , 1H), 7.32 (d, 1H);

Ms 443 (M ⁺ ).

<Manufacture example 8> Preparation of 3-carbamoyl-α-bromoacetophenone

The compound obtained in Preparation Example 1, 3-carbamoyl acetophenone (500 mg, 3.06 mmol) was dissolved in acetic acid (45 ml) and Br ₂ (0.15 ml, 3.06 mmol) was slowly added at 60 ° C. for 2.5 hours. Stirred. Acetic acid was evaporated under reduced pressure, washed with CH ₂ Cl ₂ and diethyl ether, and then recrystallized with acetonitrile (CH ₃ CN) to obtain the title compound (420 mg, 57%) as a white solid.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 4.92 (s, 2H), 5.99 (bs, 1H), 6.27 (bs, 1H), 7.64 (m, 1H), 8.10 (m, 2H), 8.43 (s , 1H);

Ms 243 (M ⁺ ).

<Manufacture example 9> Preparation of 4-carbamoyl-α-bromoacetophenone

The compound obtained in Preparation Example 2, 4-carbamoyl acetophenone was reacted in the same manner as in Preparation Example 8, to obtain the title compound (684 mg, 92%) as a pale yellow solid.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 4.96 (s, 2H), 7.58 (s, 1H), 8.02 (m, 4H), 8.16 (s, 1H);

Ms 243 (M ⁺ ).

Production Example 10 Preparation of 3- (pyridin-2-yl) -α-bromoacetophenone

The compound obtained in Preparation Example 3, 3- (pyridin-2-yl) acetophenone (2.15 g, 10.9 mmol) was dissolved in HBr (25 ml), and Br ₂ (0.56 ml, 10.9 mmol) was added at room temperature. And stirred at 70 ° C. for 12 h. Cooled to room temperature, extracted with ethyl acetate (150 ml) and washed with water and brine. The extract was dried over anhydrous MgSO ₄ , filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (hexane: ethyl acetate = 4: 1, v / v) to give the title compound (410 mg, 10%) as a white solid. Got it.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 4.55 (s, 2H), 7.30 (m, 1H), 7.61 (m, 1H), 7.81 (m, 1H), 8.03 (d, 1H), 8.27 (d , 1H), 8.61 (s, 1H), 8.73 (m, 1H);

Ms 275 (M ⁺ ).

Production Example 11 Preparation of 4- (chlorophenyl-4-yl) -α-bromoacetophenone

The compound obtained in Preparation Example 4, 4- (4-chlorophenyl-4-yl) acetophenone (70 mg, 0.3 mmol) was dissolved in diethyl ether (2 ml), and Br ₂ (15 μl, 0.3 mmol) was added thereto. It was added at room temperature. After stirring for 1.5 hours at room temperature, water (1 ml) was added to terminate the reaction, extracted with ethyl acetate (15 ml) and washed with water and brine. The extract was dried over anhydrous MgSO ₄ , filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (hexane: ethyl acetate = 15: 1, v / v) to obtain the target compound (89 mg, 96%) as a white solid. Got it.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 4.47 (s, 2H), 7.46 (dd, 2H), 7.55 (dd, 2H), 7.69 (d, 2H), 8.08 (d, 2H);

Ms 310 (M ⁺ ).

Production Example 12 Preparation of 4- (pyrrolidin-1-yl) -α-bromoacetophenone

Step 1. 4- ( Pyrrolidine -1-yl) -α, α- Of dibromoacetophenone  Produce

The compound obtained in Preparation Example 5, 4- (pyrrolidin-1-yl) acetophenone (325 mg, 1.71 mmol) was dissolved in H ₂ SO ₄ (2.5 ml) and Br ₂ (88 μl, 1.71 at 0 ° C.). mmol) was added slowly and stirred at room temperature for 3 hours. After the reaction was completed, ice water was poured into the reaction solution to form a solid, and the resultant was filtered. The filtered solid compound was washed several times with water and dried to obtain the target compound (326 mg, 55%) as a green solid.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 2.00 (m, 4H), 3.40 (m, 4H), 6.57 (d, 2H), 7.80 (s, 1H), 7.89 (d, 2H).

Step 2. 4- ( Pyrrolidine -1-yl) -α- Of bromoacetophenone  Produce

4- (pyrrolidin-1-yl) -α, α-dibromoacetophenone (320 mg, 0.92 mmol) obtained in step 1 was dissolved in THF (2 ml) and diethylphosphite (0.13 ml, 1.01 mmol) and triethylamine (0.14 ml, 1.01 mmol) were dissolved in THF (1.5 ml) and slowly added to the reaction solution at room temperature. After stirring for 4 hours, the mixture was extracted with ethyl acetate (20 ml) and washed with water and brine. The extract was dried over anhydrous MgSO ₄ , filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (hexane: ethyl acetate = 9: 1, v / v) to give the target compound (208 mg, 85%) as a brown solid. Got it.

¹ H-NMR (300 MHz, CDCl ₃ ) α 2.06 (m, 4H), 3.41 (m, 4H), 4.38 (s, 2H), 6.55 (d, 2H), 7.91 (d, 2H);

Ms 267 (M ⁺ ).

Production Example 13 Preparation of 4- (piperidin-1-yl) -α-bromoacetophenone

Step 1. 4- (piperidin-1-yl) -α, α- Of dibromoacetophenone  Produce

Using the compound obtained in Preparation Example 6, 4- (piperidin-1-yl) acetophenone was carried out in the same manner as in Step 1 of Preparation Example 12, to obtain the target compound (432 mg, 70%). .

¹ H-NMR (300 MHz, CDCl ₃ ) σ 1.69 (m, 6H), 3.44 (m, 4H), 6.67 (s, 1H), 6.85 (d, 2H), 7.98 (d, 2H);

Ms 362 (M + l) ⁺ .

Step 2. 4- (Piperidin-1-yl) -α- Of bromoacetophenone  Produce

The reaction was carried out in the same manner as in Step 2 of Preparation Example 12 using 4- (piperidin-1-yl) -α, α-dibromoacetophenone obtained in Step 1 to obtain the target compound (234 mg, 90%).

¹ H-NMR (300 MHz, CDCl ₃ ) σ 1.67 (m, 6H), 3.41 (m, 4H), 4.36 (s, 2H), 6.85 (d, 2H, J = 8.7 Hz), 7.88 (d, 2H , J = 7.5 Hz);

Ms 284 (M + 1) ⁺ .

Production Example 14 Preparation of 3- (pyrrolidin-1-yl) -α-bromoacetophenone

Step 1. 3- ( Pyrrolidine -1-yl) -α, α- Of dibromoacetophenone  Produce

Using the compound obtained in Preparation Example 7, 3- (pyrrolidin-1-yl) acetophenone, the reaction was carried out in the same manner as in Step 1 of Preparation Example 12, to obtain the target compound (534 mg, 90%). .

¹ H-NMR (300 MHz, CDCl ₃ ) σ 2.04 (m, 4H), 6.76 (s, 1H), 6.81 (d, 2H), 7.20 (s, 1H) 7.28 (m, 2H).

Step 2. 3- ( Pyrrolidine -1-yl) -α- Of bromoacetophenone  Produce

The reaction was carried out in the same manner as in Step 2 of Preparation Example 12 using 3- (pyrrolidin-1-yl) -α, α-dibromoacetophenone obtained in Step 1 to obtain the target compound (119). mg, 48%).

¹ H-NMR (300 MHz, CDCl ₃ ) σ 2.03 (m, 4H), 3.33 (m, 4H), 4.46 (s, 2H), 6.79 (dd, 1H), 7.14 (s, 1H), 7.21 (d , 1H), 7.31 (dd, 1H);

Ms 270 (M + 1) ⁺ .

Preparation Example 15 Preparation of N- (3-chloro-4-methoxyphenyl) thiourea

Benzoyl isocyanate (2 g, 12.2 mmol) was dissolved in acetone (30 ml) and 3-chloro-p-anisidine (1.92 g, 12.2 mmol) was added at room temperature. After stirring for 5 hours at room temperature, acetone was concentrated under reduced pressure to obtain a white solid compound, N- (3-chloro-4-methoxyphenyl) -N'-benzoylthiourea. The obtained N- (3-chloro-4-methoxyphenyl) -N'-benzoylthiouree was dissolved in a mixed solvent of THF (15 ml) and MeOH (15 ml), and 2N NaOH (18.3 ml, 36.6 mmol) was added at room temperature. After stirring for 2 hours at room temperature, 2N HCl was added to neutralize and the mixed solvent of THF and MeOH was evaporated under reduced pressure. After evaporation, the residue was extracted with ethyl acetate (80 ml) and washed with water and brine. The extract was dried over anhydrous MgSO ₄ , filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (hexane: ethyl acetate = 1: 2, v / v) to give the title compound (2.31 g, 87%) as a white solid. Got it.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 3.83 (s, 3H), 7.08 (d, 1H), 7.23 (dd, 1H), 7.50 (m, 3H), 9.55 (s, 1H);

Ms 216 (M ⁺ ).

2. Preparation of Compounds of the Invention

Example 1 Preparation of 2- (3-chloro-4-methoxyanilino) -4-phenylthiazole

α-bromoacetophenone (200 mg, 1.0 mmol) was dissolved in ethanol (4 ml) and NaSCN (97 mg, 1.2 mmol) was added at room temperature. The reaction was stirred at 60 ° C. for 3 hours, after which 3-chloro-p-anisidine (168 mg, 1.08 mmol) was added. After heating to reflux for 12 hours, the reaction was terminated by adding water (2 ml), extracted with ethyl acetate (20 ml) and washed with water and brine. The extract was dried over anhydrous MgSO ₄ , filtered and concentrated under reduced pressure, and purified by silica gel column chromatography (hexane: ethyl acetate = 6: 1, v / v) to give the title compound (146 mg, 46%) as a yellow solid. Got it.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 3.89 (s, 3H), 6.79 (s, 1H), 6.89 (d, 1H), 7.32 (m, 5H), 7.52 (s, 1H), 7.95 (m , 2H);

Ms 316 (M ⁺ ).

Example 2 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (3-bromophenyl) thiazole

The reaction was carried out in the same manner as in Example 1 using 2,3'-dibromoacetophenone to obtain the target compound (238 mg, 60%).

¹ H-NMR (300 MHz, CDCl ₃ ) σ 3.89 (s, 3H), 6.78 (s, 1H), 6.90 (d, 1H), 7.25 (m, 2H), 7.38 (m, 2H), 7.69 (dd , 1H), 7.73 (s, 1H), 7.95 (dd, 1H);

Ms 396 (M ⁺ ).

Example 3 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (3-carbamoylphenyl) thiazole

3-carbamoyl-α-bromoacetophenone (100 mg, 0.41 mmol) obtained in Preparation Example 8 was dissolved in ethanol (3 ml), and the compound obtained in Preparation Example 15, N- (3-chloro-4-meth Toxyphenyl) thiourea (64 mg, 0.41 mmol) was added at room temperature and then heated to reflux for 4 hours. Cooled to room temperature, extracted with ethyl acetate and washed with water and brine. The extract was dried over anhydrous MgSO ₄ , filtered and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (hexane: ethyl acetate = 4: 1, v / v) to give the title compound as a white solid (91 mg, 62). %) Was obtained.

¹ H-NMR (300 MHz, DMSO- d6 ) σ 3.90 (s, 3H), 7.15 (d, 1H), 7.39 (dd, 2H), 7.51 (dd, 1H), 7.62 (dd, 1H), 7.78 ( d, 1H), 8.02 (d, 2H), 8.37 (s, 1H), 10.27 (s, 1H);

Ms 359 (M ⁺ ).

Example 4 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (3-nitrophenyl) thiazole

The reaction was carried out in the same manner as in Example 3 using 3′-nitro-α-bromoacetophenone to give the title compound (145 mg, 98%) as a yellow solid.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 4.30 (s, 3H), 7.15 (m, 1H), 7.48 (m, 1H), 7.73 (m, 2H), 8.24 (m, 3H), 8.75 (s, 1 H), 10.47 (s, 1 H);

Ms 361 (M ⁺ ).

Example 5 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (3-aminophenyl) thiazole

2- (3-chloro-4-methoxyanilino) -4- (3-nitrophenyl) thiazole (1.1 g, 3.04 mmol) obtained in Example 4 was diluted with methanol (12 ml) and THF (12 ml). After dissolved in a mixed solvent of 50% Raney Ni (550 mg) was added at room temperature, and reacted for 2 hours in a hydrogen gas environment of 50 psi pressure. The reaction mixture was filtered through celite and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane: ethyl acetate = 2: 1, v / v) to give the title compound (900 mg, 90%) as a brown solid.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 3.83 (s, 3H), 5.10 (s, 2H), 6.53 (m, 1H), 7.09 (m, 5H), 7.63 (dd, 1H), 7.82 (s, 1 H), 10.15 (s, 1 H);

Ms 331 (M ⁺ ).

Example 6 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (3- (pyrrolidin-2-yl) phenyl) thiazole

The reaction was carried out in the same manner as in Example 3 using 3- (pyrrolidin-1-yl) -α-bromoacetophenone obtained in Preparation Example 14, to obtain the target compound as a brown solid (77 mg, 49% )

¹ H-NMR (300 MHz, CDCl ₃ ) σ 2.00 (m, 4H), 3.35 (m, 4H), 3.90 (s, 3H), 6.52 (d, 1H), 6.77 (s, 1H), 6.93 (d , 1H), 7.09 (m, 2H), 7.22 (m, 3H), 7.60 (d, 1H);

Ms 385 (M ⁺ ).

Example 7 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (3- (pyrrole-1-yl) phenyl) thiazole

2- (3-chloro-4-methoxyanilino) -4- (3-aminophenyl) thiazole (70 mg, 0.21 mmol) obtained in Example 5 was dissolved in acetic acid (1.5 ml), and 2, 5-dimethoxy tetrahydrofuran (35 μl, 0.27 mmol) was added and then stirred at 60 ° C. for 4 hours. After the reaction was terminated, extracted with ethyl acetate (20 ml) and washed with water and brine. The extract was dried over anhydrous MgSO ₄ , filtered and concentrated under reduced pressure to be purified by silica gel column chromatography (hexane: ethyl acetate = 4: 1, v / v) to give the title compound as a red solid (34 mg, 45%). Got.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 3.89 (s, 3H), 6.36 (m, 2H), 6.80 (s, 1H), 6.93 (d, 1H), 7.25 (d, 2H), 7.34 (m , 5H), 7.65 (d, 1 H), 7.85 (s, 1 H);

Ms 381 (M ⁺ ).

Example 8 Preparation of 2- (3-chloro-4-methoxyanilino) -5- (3- (piperidin-yl) phenyl) thiazole

2- (3-chloro-4-methoxyanilino) -4- (3-aminophenyl) thiazole (50 mg, 0.15 mmol) obtained in Example 5 was dissolved in water (1.5 ml) and carbonated at room temperature. Potassium (K ₂ CO ₃ ; 41 mg, 0.3 mmol) and 1,5-dibromopentane (25 μl, 0.18 mmol) were added and then stirred using microwave at 100 ° C. for 30 minutes. After cooling to room temperature, the mixture was extracted with ethyl acetate (20 ml) and washed with water and brine. The extract was dried over anhydrous MgSO ₄ , filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (hexane: ethyl acetate = 4: 1, v / v) to give the target compound (25 mg, 42%) in oil form. Got it.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 1.25 (m, 2H), 1.75 (m, 4H), 3.21 (m, 4H), 3.88 (s, 3H), 6.75 (s, 1H), 6.94 (m , 2H), 7.21 (m, 3H), 7.51 (d, 2H);

Ms 399 (M ⁺ ).

Example 9 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (3- (pyridin-2-yl) phenyl) thiazole

The reaction was carried out in the same manner as in Example 3 using 3- (pyridin-2-yl) -α-bromoacetophenone obtained in Preparation Example 10 to obtain the target compound (48 mg, 30%) in oil form. .

¹ H-NMR (300 MHz, CDCl ₃ ) σ 3.89 (s, 3H), 6.92 (m, 2H), 7.29 (m, 3H), 7.53 (m, 2H), 7.77 (m, 2H), 7.89 (m , 2H), 8.45 (s, 1 H), 8.68 (d, 1 H);

Ms 393 (M ⁺ ).

Example 10 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (3- (3-bromopropionamido) phenyl) thiazole

2- (3-chloro-4-methoxyanilino) -4- (3-aminophenyl) thiazole (155 mg, 0.47 mmol) obtained in Example 5 was dissolved in THF (2 ml) and triethyl Amine (65 μl, 0.47 mmol) was added at room temperature followed by the slow addition of 3-bromopropionyl chloride (96 μl, 0.95 mmol). After stirring for 4 hours, water (1 ml) was added to the reaction mixture, which was extracted with ethyl acetate (30 ml) and washed with water and brine. The extract was dried over anhydrous MgSO ₄ , filtered, and concentrated under reduced pressure, and purified by silica gel column chromatography (hexane: ethyl acetate = 2: 1, v / v) to give the title compound (110 mg, 50%) as a white solid. Got it.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 2.98 (t, 2H), 3.76 (t, 2H), 3.83 (s, 1H), 7.17 (d, 1H), 7.21 (s, 1H), 7.36 (m, 1H), 7.57 (m, 2H), 7.65 (dd, 1H), 7.81 (d, 1H), 8.13 (s, 1H), 10.14 (s, 1H), 10.22 (s, 1H);

Ms 467 (M ⁺ ).

Example 11 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (3-acrylamidophenyl) thiazole

2- (3-chloro-4-methoxyanilino) -4- (3- (3-bromopropionamido) phenyl) thiazole (100 mg, 0.21 mmol) obtained in Example 10 was diluted with THF (2 ml) and triethylamine (100 μl, 0.59 mmol) were added at room temperature and then heated to reflux for 12 hours. After the reaction was terminated and extracted with ethyl acetate (15 ml) and washed with water and brine. The extract was dried over anhydrous MgSO ₄ , filtered and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (hexane: ethyl acetate = 1: 1, v / v) to give the target compound as an oil (65 mg, 80%).

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 3.83 (s, 3H), 5.76 (dd, 1H), 6.31 (dd, 1H), 6.43 (dd, 1H), 7.17 (d, 1H), 7.22 (s, 1H), 7.38 (m, 1H), 7.61 (m, 3H), 7.80 (d, 1H), 8.17 (s, 1H), 10.21 (s, 1H), 10.22 (s, 1H);

Ms 385 (M ⁺ ).

Example 12 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (3-methanesulfonamidophenyl) thiazole

2- (3-chloro-4-methoxyanilino) -4- (3-aminophenyl) thiazole (100 mg, 0.3 mmol) obtained in Example 5 was dissolved in CH ₂ Cl ₂ (2 ml) Triethylamine (50 μl, 0.36 mmol) was added at 0 ° C., then methanesulfonyl chloride (28 μl, 0.36 mmol) was added slowly. After stirring for 2 hours at room temperature, the mixture was extracted with CH ₂ Cl ₂ (20 ml) and washed with water and brine. The extract was dried over anhydrous MgSO ₄ , filtered, and concentrated under reduced pressure and purified by silica gel column chromatography (hexane: ethyl acetate = 2: 1, v / v) to give the title compound as a pale yellow solid (44 mg, 36%). Got.

¹ H-NMR (300 MHz, DMSO- d ₆ ) δ 3.03 (s, 3H), 3.83 (s, 3H), 7.12 (m, 2H), 7.27 (s, 1H), 7.38 (dd, 1H), 7.63 (m, 2H), 7.78 (s, 1H), 7.83 (d, 1H), 9.83 (s, 1H), 10.24 (s, 1H);

Ms 409 (M ⁺ ).

Example 13 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (3-dimethylsulfonamidophenyl) thiazole

The reaction was carried out in the same manner as in Example 12 using methanesulfonyl chloride (56 μl, 0.72 mmol) to obtain the target compound (14 mg, 10%) as a light red solid.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 3.58 (s, 6H), 3.83 (s, 3H), 7.14 (d, 1H), 7.54 (m, 4H), 7.95 (s, 1H), 7.98 (d, 1H), 8.02 (d, 1H), 10.31 (s, 1H);

Ms 487 (M ⁺ ).

Example 14 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (3-hydroxyphenyl) thiazole

The reaction was carried out in the same manner as in Example 3 using 3'-hydroxy-α-bromoacetophenone to give the title compound (111 mg, 82%) as a white solid.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 3.82 (s, 3H), 6.58 (d, 1H), 7.05 (m, 5H), 7.14 (d, 1H), 7.69 (d, 1H), 10.02 (s, 1 H);

Ms 332 (M ⁺ ).

Example 15 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (3- (2- (pyrrolidin-1-yl) ethoxy) phenyl) thiazole

2- (3-chloro-4-methoxyanilino) -4- (3-hydroxyphenyl) thiazole (100 mg, 0.3 mmol) obtained in Example 14 was dissolved in DMF (1.5 ml) and K ₂ CO ₃ (124 mg, 0.9 mmol) was added followed by 1- (2-chloroethyl) pyrrolidine hydrochloride (61 mg, 0.36 mmol). After stirring for 12 hours at 120 ℃, cooled to room temperature, extracted with ethyl acetate (20 ml) and washed with water and brine. The extract was dried over anhydrous MgSO ₄ , filtered, and concentrated under reduced pressure and purified by silica gel column chromatography (5% MeOH in CH ₂ Cl ₂ ) to obtain the title compound (44 mg, 34%) as a white solid.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 1.83 (m, 4H), 2.72 (m, 4H), 2.98 (t, 2H), 3.88 (s, 3H), 4.20 (t, 2H), 6.61 (s , 1H), 6.84 (m, 2H), 7.27 (m, 2H), 7.49 (m, 2H), 7.56 (d, 1H);

Ms 429 (M ⁺ ).

Example 16 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (3- (2- (piperidin-1-yl) ethoxy) phenyl) thiazole

The above procedure was carried out using 2- (3-chloro-4-methoxyanilino) -4- (3-hydroxyphenyl) thiazole and 1- (2-chloroethyl) piperidine hydrochloride obtained in Example 14. The reaction was carried out in the same manner as in Example 15, to obtain the target compound (36 mg, 27%) as a yellow solid.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 1.46 (m, 2H), 1.61 (m, 4H), 2.53 (m, 4H), 2.80 (t, 2H), 3.89 (s, 3H), 4.16 (t , 2H), 6.76 (s, 1H), 6.85 (dd, 1H), 6.91 (d, 1H), 7.27 (m, 2H), 7.40 (m, 2H), 7.51 (d, 1H);

Ms 443 (M ⁺ ).

Example 17 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (3- (2- (morpholin-1-yl) ethoxy) phenyl) thiazole

Example 2 using 2- (3-chloro-4-methoxyanilino) -4- (3-hydroxyphenyl) thiazole and 1- (2-chloroethyl) morpholine hydrochloride obtained in Example 14 above The reaction was performed in the same manner as 15 to obtain the target compound (44 mg, 33%) as a brown solid.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 2.59 (m, 4H), 2.82 (t, 2H), 3.72 (m, 4H), 3.89 (s, 3H), 4.17 (t, 2H), 6.77 (s , 1H), 6.84 (dd, 1H), 6.89 (d, 1H), 7.27 (m, 2H), 7.41 (m, 1H), 7.53 (d, 1H);

Ms 445 (M ⁺ ).

Example 18 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4-chlorophenyl) thiazole

The reaction was carried out in the same manner as in Example 1 using 4'-chloro-α-bromoacetophenone to give the title compound (221 mg, 63%) as a yellow solid.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 3.90 (s, 3H), 6.77 (s, 1H), 6.91 (d, 1H), 7.33 (m, 4H), 7.37 (d, 1H), 7.76 (dd , 2H);

Ms 351 (M ⁺ ).

Example 19 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4-fluorophenyl) thiazole

The reaction was carried out in the same manner as in Example 1 using 4'-fluoro-α-bromoacetophenone to give the title compound (97 mg, 29%) as a brown solid.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 3.96 (s, 3H), 6.71 (s, 1H), 6.94 (d, 1H), 7.10 (m, 3H), 7.31 (d, 1H), 7.48 (d, 1 H), 7.80 (m, 2 H);

Ms 334 (M ⁺ ).

Example 20 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4-cyanophenyl) thiazole

The reaction was carried out in the same manner as in Example 1 using 4'-cyano-α-bromoacetophenone to give the title compound (331 mg, 97%) as a yellow solid.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 7.00 (m, 2H), 7.49 (m, 3H), 7.75 (m, 1H), 7.95 (d, 2H), 8.33 (d, 1H), 11.74 (s, 1 H);

Ms 341 (M ⁺ ).

Example 21 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4-bromophenyl) thiazole

The reaction was carried out in the same manner as in Example 3 using 2,4'-dibromoacetophenone (600 mg) to obtain a target compound (143 mg, 88%) as a yellow solid.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 3.90 (s, 3H), 6.76 (s, 1H), 6.92 (d, 1H), 7.24 (m, 1H), 7.47 (m, 3H), 7.68 (d , 2H);

Ms 396 (M ⁺ ).

Example 22 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4-methylphenyl) thiazole

The reaction was carried out in the same manner as in Example 1 using 4'-methyl-α-bromoacetophenone to give the title compound (115 mg, 26%) as a white solid.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 3.82 (s, 3H), 7.14 (dd, 1H), 7.40 (s, 1H), 7.64 (dd, 1H), 7.81 (m, 5H), 10.38 (s, 1 H);

Ms 442 (M ⁺ ).

Example 23 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4-trifluoromethylphenyl) thiazole

The reaction was performed in the same manner as in Example 1 using 4'-trifluoromethyl-α-bromoacetophenone to give the title compound (361 mg, 94%) as a yellow solid.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 3.83 (s, 3H), 7.18 (d, 1H), 7.57 (s, 1H), 7.64 (dd, 1H), 7.78 (d, 2H), 7.86 (d, 1H), 8.12 (d, 2H), 10.30 (s, 1H);

Ms 384 (M ⁺ ).

Example 24 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4-iodophenyl) thiazole

The reaction was carried out in the same manner as in Example 3 using 4'-iodo-α-bromoacetophenone to give the title compound (161 mg, 89%) as a brown solid.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 3.82 (s, 3H), 7.14 (dd, 1H), 7.40 (s, 1H), 7.64 (dd, 1H), 7.81 (m, 5H), 10.38 (s, 1 H);

Ms 442 (M ⁺ ).

Example 25 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4-carbamoylphenyl) thiazole

The reaction was carried out in the same manner as in Example 3 using 4-carbamoyl-α-bromoacetophenone obtained in Preparation Example 9 to obtain the target compound (132 mg, 90%) as a white solid.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 3.99 (s, 3H), 7.32 (m, 1H), 7.66 (m, 2H), 7.80 (dd, 1H), 7.96 (d, 1H), 8.10 (m, 5 H), 10.45 (s, 1 H);

Ms 359 (M ⁺ ).

Example 26 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4-nitrophenyl) thiazole

The reaction was carried out in the same manner as in Example 3 using 4'-nitro-α-bromoacetophenone to give the title compound (145 mg, 98%) as a yellow solid.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 3.83 (s, 3H), 7.19 (d, 1H), 7.65 (dd, 1H), 7.72 (s, 1H), 7.83 (d, 1H), 8.13 (d, 2H), 8.32 (d, 2H). 10.36 (s, 1 H);

Ms 361 (M ⁺ ).

Example 27 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4-aminophenyl) thiazole

2- (3-chloro-4-methoxyanilino) -4- (4-nitrophenyl) thiazole (1 g, 2.7 mmol) obtained in Example 26 was dissolved in methanol (9 ml) and 0.36 at room temperature. M aqueous Cu (OAc) ₂ solution (7.6 ml, 2.7 mmol) was added followed by the slow addition of NaBH ₄ (1.2 g, 32.4 mmol) at 0 ° C. After stirring for 5 minutes at room temperature, methanol was evaporated under reduced pressure, extracted with ethyl acetate (40 ml) and washed with water and brine. The extract was dried over MgSO ₄ , filtered, and concentrated under reduced pressure. Got it.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 3.82 (s, 3H), 5.23 (s, 2H), 6.61 (d, 2H), 6.88 (s, 1H), 7.17 (d, 1H), 7.58 (m , 3H), 7.88 (d, 1 H), 10.10 (s, 1 H);

Ms 331 (M ⁺ ).

Example 28 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4-N-methylaminophenyl) thiazole

2- (3-chloro-4-methoxyanilino) -4- (4-aminophenyl) thiazole (100 mg, 0.3 mmol) obtained in Example 27 was dissolved in DMF (1.5 ml) and K was added at room temperature. ₂ CO ₃ (104 mg, 0.75 mmol) was added followed by slow addition of iodomethane (41 μl, 0.66 mmol). After stirring for 8 hours at room temperature, water (1 ml) was added to terminate the reaction, extracted with ethyl acetate (20 ml) and washed with water and brine. The extract was dried over anhydrous MgSO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane: ethyl acetate = 4: 1, v / v) to give the title compound (15 mg, 15%) as a purple solid.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 2.81 (s, 3H), 3.86 (s, 3H), 6.53 (s, 1H), 6.60 (d, 2H), 6.94 (d, 1H), 7.24 (m , 2H), 7.48 (d, 1H), 7.66 (d, 2H);

Ms 345 (M ⁺ ).

Example 29 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4-N, N-dimethylaminophenyl) thiazole

The reaction was carried out in the same manner as in Example 28 using iodo methane (82 μl, 1.32 mmol) to obtain the target compound (18 mg, 17%) as a purple solid.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 2.97 (s, 6H), 3.92 (s, 3H), 6.56 (s, 1H), 6.73 (d, 2H), 6.90 (d, 1H), 7.25 (m , 2H), 7.45 (d, 1H), 7.68 (m, 2H);

Ms 359 (M ⁺ ).

Example 30 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4-N-1-butylaminophenyl) thiazole

2- (3-chloro-4-methoxyanilino) -4- (4-aminophenyl) thiazole (100 mg, 0.3 mmol) obtained in Example 27 was dissolved in CH ₂ Cl ₂ (1.5 ml), and Titanium isopropoxide (0.11 ml, 0.38 mmol) and n-butylaldehyde (27 μl) were added at room temperature, followed by stirring at room temperature for 2 hours, cooling to 0 ° C. to give NaBH ₄ (34.2 mg, 0.9 mmol). Added slowly. After stirring for 1 hour at room temperature, extracted with CH ₂ Cl ₂ (10 ml) and washed with water and brine. The extract was dried over anhydrous MgSO ₄ , filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (hexane: ethyl acetate = 2: 1, v / v) to give a red solid target compound (32 mg, 28%). Got.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 0.97 (t, 3H), 1.40 (m, 2H), 1.64 (m, 2H), 3.14 (m, 2H), 3.87 (m, 1H), 3.89 (s , 3H), 6.50 (s, 1H), 6.59 (d, 2H), 6.92 (d, 2H), 7.28 (d, 1H), 7.47 (d, 1H), 7.65 (d, 2H);

Ms 387 (M ⁺ ).

Example 31 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4- (pyrrolidin-1-yl) phenyl) thiazole

The reaction was carried out in the same manner as in Example 3 using 3- (pyrrolidin-1-yl) -α-bromoacetophenone obtained in Preparation Example 14, to obtain the target compound as a red solid (128 mg, 81 %) Was obtained.

¹ H-NMR (300 MHz, DMSO - d ₆ ) σ 1.92 (m, 4H), 3.82 (s, 3H), 5.30 (m, 4H), 6.63 (d, 2H), 6.97 (s, 1H), 7.17 (d, 1H), 7.56 (m, 1H), 7.70 (d, 2H), 7.93 (d, 1H), 10.20 (s, 1H);

Ms 385 (M ⁺ ).

Example 32 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4- (piperidin-1-yl) phenyl) thiazole

The reaction was carried out in the same manner as in Example 3 using 4- (piperidin-1-yl) -α-bromoacetophenone obtained in Preparation Example 13, to obtain the target compound as a light brown solid (129 mg, 79%).

¹ H-NMR (300 MHz, DMSO - d ₆ ) σ 1.79 (m, 6H), 3.53 (m, 4H), 3.83 (s, 3H), 7.13 (d, 1H), 7.41 (s, 1H), 7.59 (m, 4H), 7.93 (m, 2H), 10.40 (s, 1H);

Ms 399 (M ⁺ ).

Example 33 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4- (pyrrole-1-yl) phenyl) thiazole

2- (3-chloro-4-methoxyanilino) -4- (4-aminophenyl) thiazole (150 mg, 0.45 mmol) obtained in Example 27 was dissolved in dioxane (2 ml) and stored at room temperature. 2,5-dimethoxy tetrahydrofuran (76 μl, 0.58 mmol) was added, followed by 4-chloropyridine hydrochloride (7 mg, 0.045 mmol) and heated to reflux for 6 hours. After cooling to room temperature, water (1 ml) was added to terminate the reaction, and extracted with ethyl acetate (20 ml) and washed with water and brine. The extract was dried over anhydrous MgSO ₄ , filtered, and concentrated under reduced pressure, and purified by silica gel column chromatography (hexane: ethyl aceteat = 4: 1, v / v) to give a brown solid target compound (32 mg, 19%). )

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 3.81 (s, 3H), 6.28 (m, 2H), 7.15 (d, 1H), 7.35 (s, 1H), 7.44 (d, 2H), 7.59 (m, 3 H), 7.95 (m, 3 H), 10.25 (s, 1 H);

Ms 381 (M ⁺ ).

Example 34 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4- (3-bromopropionamido) phenyl) thiazole

The reaction was carried out in the same manner as in Example 10, using 2- (3-chloro-4-methoxyanilino) -4- (4-aminophenyl) thiazole obtained in Example 27 to obtain a dark yellow solid. The desired compound (178 mg, 81%) was obtained.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 2.99 (t, 2H), 3.74 (t, 2H), 3.82 (s, 3H), 7.15 (d, 1H), 7.21 (s, 1H), 7.64 (m, 3H), 7.85 (m, 3H), 10.17 (s, 1H), 10.22 (s, 1H);

Ms 467 (M ⁺ ).

Example 35 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4-acrylamidophenyl) thiazole

In the same manner as in Example 11 using 2- (3-chloro-4-methoxyanilino) -4- (4- (3-bromopropionamido) phenyl) thiazole obtained in Example 34 The reaction was carried out to obtain the target compound (61 mg, 75%) in oil form.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 3.88 (s, 3H), 5.81 (dd, 1H), 6.36 (d, 1H), 6.50 (m, 1H), 7.24 (d, 1H), 7.28 (s, 1H), 7.72 (m, 3H), 7.93 (m, 3H), 10.26 (d, 2H);

Ms 385 (M ⁺ ).

Example 36 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4-hydroxyphenyl) thiazole

The reaction was carried out in the same manner as in Example 3 using 4'-hydroxy-α-bromoacetophenone to give the title compound (133 mg, 98%) as a yellow solid.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 3.82 (s, 3H), 6.82 (m, 2H), 7.16 (m, 2H), 7.51 (d, 1H), 7.73 (m, 3H), 10.29 (s, 1 H);

Ms 332 (M ⁺ ).

Example 37 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4- (2- (piperidin-1-yl) ethoxy) phenyl) thiazole

Example 2 using 2- (3-chloro-4-methoxyanilino) -4- (4-hydroxyphenyl) thiazole and 1- (2-chloroethyl) piperidine hydrochloride obtained in Example 36 The reaction was performed in the same manner as 15 to obtain the target compound (36 mg, 27%) as a brown solid.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 1.27 (m, 9H), 2.47 (m, 2H), 3.58 (s, 3H), 3.85 (m, 3H), 6.74 (d, 2H), 6.90 (d, 2H), 7.34 (m, 1H), 7.55 (d, 2H), 7.65 (d, 1H), 9.95 (s, 1H);

Ms 443 (M ⁺ ).

Example 38 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4- (2- (pyrrolidin-1-yl) ethoxy) phenyl) thiazole

Example 2 using 2- (3-chloro-4-methoxyanilino) -4- (4-hydroxyphenyl) thiazole and 1- (2-chloroethyl) pyrrolidine hydrochloride obtained in Example 36 The reaction was performed in the same manner as 15 to obtain the target compound (26 mg, 20%) as a brown solid.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 1.86 (m, 4H), 2.70 (m, 4H), 3.01 (t, 2H), 3.89 (s, 3H), 4.19 (t, 2H), 6.63 (s , 1H), 6.93 (d, 3H), 7.31 (dd, 1H), 7.49 (d, 1H), 7.74 (d, 2H);

Ms 429 (M ⁺ ).

Example 39 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4- (2- (morpholin-1-yl) ethoxy) phenyl) thiazole

Example 15 using 2- (3-chloro-4-methoxyanilino) -4- (4-hydroxyphenyl) thiazole and 1- (2-chloroethyl) morpholine hydrochloride obtained in Example 36. The reaction was carried out in the same manner as to obtain the target compound (47 mg, 35%) as a brown solid.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 2.60 (m, 4H), 2.82 (t, 2H), 3.76 (m, 4H), 3.89 (s, 3H), 4.16 (t, 2H), 6.64 (s, 1H), 6.92 (d, 3H), 7.31 (m, 1H), 7.47 (d, 1H), 7.74 (dd, 2H), 8.02 (s, 1H);

Ms 445 (M ⁺ ).

Example 40 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4- (3- (piperidin-1-yl) propoxy) phenyl) thiazole

Example 2 using 2- (3-chloro-4-methoxyanilino) -4- (4-hydroxyphenyl) thiazole and 1- (3-chloropropyl) piperidine hydrochloride obtained in Example 36 The reaction was performed in the same manner as 15 to obtain the target compound (26 mg, 19%) as a brown solid.

¹ H NMR (300 MHz, DMSO- d6 ) σ 1.38 (m, 7H), 1.78 (m, 3H), 3.21 (m, 6H), 3.89 (m, 3H), 6.81 (d, 2H), 7.01 (d , 2H), 7.43 (dd, 1H), 7.64 (d, 2H), 7.79 (d, 1H), 10.20 (s, 1H);

Ms 457 (M ⁺ ).

Example 41 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (2,4-dichlorophenyl) thiazole

The reaction was carried out in the same manner as in Example 1 using 2,4'-difluoro-α-bromoacetophenone to give the title compound (201 mg, 52%) as a yellow solid.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 3.89 (s, 3H), 6.83 (d, 1H), 7.00 (s, 1H), 7.18 (m, 2H), 7.30 (s, 1H), 7.34 (s , 1H), 7.71 (d, 1H), 8.38 (s, 1H);

Ms 386 (M ⁺ ).

Example 42 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4-methoxy-3-nitrophenyl) thiazole

The reaction was carried out in the same manner as in Example 3 using 4'-methoxy-3'-nitro-α-bromoacetophenone to give the title compound (147 mg, 92%) as a white solid.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 3.81 (s, 3H), 3.95 (s, 3H), 7.18 (m, 1H), 7.47 (m, 2H), 7.78 (d, 1H), 8.18 (m, 1 H), 8.38 (d, 1 H), 10.29 (s, 1 H);

Ms 391 (M ⁺ ).

Example 43 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (3-amino-4-methoxyphenyl) thiazole

2- (3-chloro-4-methoxyanilino) -4- (4-methoxy-3-nitrophenyl) thiazole (990 mg, 2.53 mmol) obtained in Example 42 was added to HCl (5 ml). Dissolved and stirred. SnCl ₂ (2.4 g, 12.7 mmol) was dissolved in HCl (4 ml) and then slowly added to the stirred reactor at 50 ° C. The mixture was heated to 100 ° C., stirred for 20 minutes, and then cooled to room temperature and filtered. The filtered solid was dissolved in water, basified with 2 N NaOH, extracted with CHCl ₃ (60 ml), and washed with water and brine. The extract was dried over anhydrous MgSO ₄ , filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (hexane: ethyl acetate = 4: 1, v / v) to give a red solid target compound (516 mg, 56%). Got.

¹ H-NMR (300 MHz, DMSO- d6 ) σ 3.77 (s, 3H), 3.81 (s, 3H), 4.75 (s, 2H), 6.81 (d, 1H), 6.91 (s, 1H), 7.11 ( m, 3H), 7.61 (dd, 1H), 7.84 (d, 1H), 10.14 (s, 1H);

Ms 361 (M ⁺ ).

Example 44 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4-bromo-3-nitrophenyl) thiazole

The reaction was carried out in the same manner as in Example 3 using 4'-bromo-3'-nitro-α-bromoacetophenone to give the title compound (168 mg, 93%) as a white solid.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 3.81 (s, 3H), 7.13 (m, 1H), 7.47 (d, 1H), 7.74 (d, 1H), 7.81 (d, 1H), 8.04 (m, 2 H), 8.48 (s, 1 H), 10.46 (s, 1 H);

Ms 441 (M ⁺ ).

Example 45 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (3-amino-4-bromophenyl) thiazole

The reaction was carried out in the same manner as in Example 43, using 2- (3-chloro-4-methoxyanilino) -4- (4-bromo-3-nitrophenyl) thiazole obtained in Example 44. To give the target compound (790 mg, 76%) as a yellow solid.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 3.81 (s, 3H), 5.35 (s, 2H), 6.97 (dd, 1H), 7.11 (d, 1H), 7.14 (s, 1H), 7.33 (m, 2 H), 7.67 (d, 1 H), 7.72 (s, 1 H), 10.17 (s, 1 H);

Ms 411 (M ⁺ ).

Example 46 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4-bromo-3- (piperidin-1-yl) phenyl) thiazole

The reaction was carried out in the same manner as in Example 8, using 2- (3-chloro-4-methoxyanilino) -4- (3-amino-4-bromophenyl) thiazole obtained in Example 45. To give the title compound (12 mg, 17%) in oil form.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 1.74 (m, 6H), 3.42 (t, 1H), 3.52 (t, 1H), 3.90 (s, 3H), 4.13 (m, 2H), 6.75 (s , 1H), 6.89 (m, 1H), 7.24 (m, 2H), 7.56 (m, 3H), 8.06 (s, 1H);

Ms 479 (M ⁺ ).

Example 47 Preparation of 2- (3-chloro-4-methoxyanilino) -4-((4-bromo-3-pyrrole-1-yl) phenyl) thiazole

The reaction was carried out in the same manner as in Example 33, using 2- (3-chloro-4-methoxyanilino) -4- (3-amino-4-bromophenyl) thiazole obtained in Example 45. To give the title compound (79 mg, 38%) as a yellow solid.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 3.90 (s, 3H), 6.34 (d, 2H), 6.79 (s, 1H), 6.93 (m, 3H), 7.28 (m, 1H), 7.43 (d , 1H), 7.62 (m, 2H), 7.77 (d, 1H);

Ms 461 (M ⁺ ).

Example 48 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4'-chlorobisphenyl-4-yl) thiazole

The reaction was carried out in the same manner as in Example 3 using 4- (chlorophenyl-4-yl) -α-bromoacetophenone obtained in Preparation Example 11 to obtain the target compound (143 mg, 82%) as a white solid. Got it.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 3.82 (s, 3H), 7.16 (m, 1H), 7.51 (m, 3H), 7.77 (m, 5H), 7.98 (m, 3H), 10.27 (s , 1H);

Ms 426 (M ⁺ ).

Example 49 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4-chloro-3-nitrophenyl) thiazole

The reaction was carried out in the same manner as in Example 3 using 4'-chloro-3'-nitro-α-bromoacetophenone to give the title compound (128 mg, 79%) as a white solid.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 3.83 (s, 3H), 7.15 (d, 1H), 7.60 (dd, 1H), 7.66 (s, 1H), 7.84 (m, 2H), 8.18 (dd, 1 H), 8.52 (d, 1 H) 10.37 (s, 1 H);

Ms 395 (M ⁺ ).

Example 50 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (3-amino-4-chlorophenyl) thiazole

The reaction was carried out in the same manner as in Example 43, using 2- (3-chloro-4-methoxyanilino) -4- (4-chloro-3-nitrophenyl) thiazole obtained in Example 49. The desired compound (482 mg, 51%) was obtained as a white solid.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 3.83 (s, 3H), 5.40 (s, 2H), 7.05 (dd, 1H), 7.14 (s, 1H), 7.16 (d, 1H), 7.22 (d, 1 H), 7.33 (s, 1 H), 7.71 (m, 2 H), 10.18 (s, 1 H);

Ms 366 (M ⁺ ).

Example 51 Preparation of 2- (3-chloro-4-methoxyanilino) -4- (4-chloro-3- (piperidin-1-yl) phenyl) thiazole

The reaction was carried out in the same manner as in Example 8 using 2- (3-chloro-4-methoxyanilino) -4- (4-chloro-3-nitrophenyl) thiazole obtained in Example 49. The desired compound (32 mg, 49%) in the form of a light yellow foam was obtained.

¹ H-NMR (300 MHz, DMSO- d ₆ ) σ 1.42 (m, 2H), 1.54 (m, 4H), 2.85 (m, 4H), 3.67 (s, 3H), 6.98 (d, 1H), 7.21 (s, 1H), 7.23 (m, 2H), 7.36 (dd, 1H), 7.56 (d, 1H), 8.00 (d, 1H), 10.12 (s, 1H);

Ms 433 (M ⁺ ).

Example 52 Preparation of 2- (3-chloro-4-methoxyanilino) -4-((4-chloro-3-pyrrole-1 yl) phenyl)) thiazole

The reaction was carried out in the same manner as in Example 33, using 2- (3-chloro-4-methoxyanilino) -4- (4-chloro-3-nitrophenyl) thiazole obtained in Example 49. To give the title compound (39 mg, 21%) as a yellow solid.

¹ H-NMR (300 MHz, CDCl ₃ ) σ 3.82 (s, 3H), 6.28 (d, 2H), 7.08 (d, 2H), 7.13 (d, 1H), 7.55 (d, 1H), 7.57 (s , 1H), 7.71 (d, 1H), 7.91 (m, 3H), 10.29 (s, 1H);

Ms 416 (M ⁺ ).

Table 1 shows the structural formulas, R ¹ and R ² of the derivatives of the present invention obtained through the above examples.

Example constitutional formula R ¹ and R ² One

H H 2

H bromo 3

H carbamo 4

H nitro 5

H amino 6

H

7

H

8

H

9

H pyridinyl 10

H-NHCO (CH ₂ ) ₂ Br 11

H -NHCOCHCH ₂ 12

H -NHSO ₂ CH ₃ 13

H -N (SO ₂ CH ₃ ) ₂ 14

H hydroxy 15

H

16

H

17

H

18

Chloro H 19

Fluoro H 20

Cyano h 21

Bromo H 22

Methyl H 23

Trifluoromethyl H 24

Iodo H 25

Cabamo H 26

Nitro H 27

Amino H 28

Methylamino H 29

Dimethylamino H 30

n-butylamino H 31

H 32

H 33

H 34

-NHCO (CH ₂ ) ₂ Br H 35

-NHCOCHCH ₂ H 36

Hydroxy H 37

H 38

H 39

H 40

H 41

Chlorochloro 42

Methoxy nitro 43

Methoxy amino 44

Bromo Nitro 45

Bromo amino 47

Bromo

46

Bromo

48

4-chlorophenyl H 49

Chloro Nitro 50

Chloroamino 51

Chloro

52

Chloro

The pharmacological action of the compounds of formula 1 according to the present invention was carried out as follows.

Experimental Example 1 RTK Enzyme Inhibitory Effect

In order to measure the RTK enzyme inhibitory effect of the 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative of the present invention, Fluorescence resonance energy transfer (FRET) or time-resolved fluorescence (TRF) method was used. The following experiment was carried out using.

Experimental Example  1-1: FRET ( Fluorescence resonance energy transfer ) Way

Tyrosine kinase activity of epidermal growth factor receptor (EGFR) and vascular endothelial growth factor receptor 2 (VEGFR-2) was measured using an AlphaScreen ^™ phospho-tyrosine-100 assay method (PerkinElmer). AlphaScreen stands for Amplified Luminescent Proximity Homogeneous Assay, and is an immunoassay method using Luminescent Oxygen Radical Channel (Warner et al., Curr. Med. Chem., 11). : 721-30, 2004).

EGFR (Sigma, St. Louse, MO, USA) and VEGFR-2 (ProQuinase, Germany) were purchased and used, and white 384 well plates (Greiner Bio-One) and biotinylated poly [ Biotinylated-poly [Glu: Tyr] (4: 1); Packard BioScience) substrates were used and coated with streptavidin (Streptavidin) to detect the degree of phosphorylation of peptide substrates. A capture buffer including a donor bead and an acceptor bead coupled with an antibody (P-Tyr-100) was added and reacted in the following three steps. In addition, the inhibitory effect was compared using Tyrphostin A51, a known RTK inhibitor. First, RTK enzyme (EGFR 5 ng / VEGFR 2.5 ng / PDGFR 10 ng / FGFR 2.5 ng) was added to each well of a 384-well plate with Tyrphostin A51 (Upstate Biotech), a known standard inhibitor, or with the compound to be tested. The reaction was carried out at room temperature for 15 minutes in a volume of μl. Second, after 15 minutes of reaction, 5 μl of biotinylated poly [glutamine: tyrosine] (4: 1) and ATP were added to each well so that the final concentrations were 5 nM and 100 μM, respectively. The reaction was advanced. At this time, the composition of the buffer used for the enzyme reaction is 50 mM Tris-HCl (pH 7.5), 5 mM MgCl ₂ , 5 mM MnCl ₂ , 2 mM DTT and 0.01% Tween-20. Third, after the enzyme reaction, 10 μl of capture buffer containing donor beads and receptor beads (20 mg / ml each) was added thereto, and the mixture was left at room temperature for 1 hour. This process takes place in the dark and stops the enzymatic reaction in progress and recognizes the phosphorylated substrate of the donor-streptavidin and the receptor-antiphosphate antibody. The composition of the capture buffer (2.5-fold concentration) is 62.5 mM HEPES (pH 7.4), 250 mM NaCl, 100 mM EDTA and BSA 0.25%. After the reaction was completed, the activity of the enzyme was determined by measuring the alpha screen signal using a Fusion ^™ microplate analyzer.

For the positive control, 1 μl of solvent DMSO was added instead of the compound, and the negative control was adjusted by adding DMSO instead of the compound and buffer instead of the enzyme. The% degradation from the measured signal was calculated as follows.

The experiment to find the concentration-response curve was conducted in triples. In other words, by diluting the inhibitor, the percent inhibition value at 6 or more concentrations was obtained, and the result obtained was nonlinear regression fitting using a program Prism ^™ (GraphPad software, San Diego, Calif.) To obtain an IC ₅₀ value.

Experimental Example  1-2: TRF ( Time - resolved fluorescence ) Way

The TRF method is a screening method that measures the difference between the excitation time and the emission time by using the property of the fluorescence emission of the lanthanide-based element, and the fluorescence of the compound itself is measured by the time difference. Interference can be eliminated (Waddleton et al., Anal. Biochem., 309: 150-7, 2002). In order to prepare a substrate having specificity for each phosphorylation enzyme to be searched, an optimal peptide substrate was found by scanning activity against a set of tyrosine substrates (145 species, 370 Tyrosine residues, JPT, Germany). Here, a peptide substrate with biotin attached to the N-terminus was synthesized (Peptron Corporation, Korea) and used for TRF search. The amino acid sequences used for the search were TLTTNEE Y LDLSQ (for EGFR search) and Ahx-EEEE Y FELVAKKKK (for VEGFR-2 search). Substrate (peptide, 1.5 μM), enzyme (30 ng), ATP (200 μM) was added to the reaction buffer (60 mM HEPES, pH 7.5, 5 mM MgCl ₂ , 5 mM MnCl ₂ , 3 μM Na ₃ VO _4). , DTT 1.25 mM) were mixed in an appropriate amount to prepare 100 μl of reaction solution. The reaction mixture was left to react for 30 minutes, and then an equal amount of reaction termination buffer (50 mM EDTA, pH 8.0) was dispensed to terminate the reaction, and the reaction mixture was transferred to a streptavidin-coated plate (DELFIA). The reaction was carried out again for 60 minutes. Phosphorylated product was bound to the plate bottom, and the rest of the reaction was removed by washing with PBST containing 0.05% Tween-20 (3 times). The prepared primary antibody (anti-phosphotyrosine monoclonal antibody) was added and reacted for 1 hour, and then the unbound primary antibody was washed with PBST buffer (repeated three times). The prepared secondary antibody (Europium-labeled anti-mouse IgG) was added and reacted for 30 minutes, and the unbound secondary antibody was washed again with PBST buffer (repeat 5 times). After the prepared IA Enhancement Solutione (PerkinElmer Co.) was added and reacted for 5 minutes, 1420 Victor 2 (VICTOR2, PerkinElmer Co.) was used to emit 615 nm Europium (Eu) emission energy in TRF mode. (delay time 400 msec). As a standard compound, PDGFR inhibitor III (PDGFR inhibitor III, Calbiochem Co., Ltd.) was used, and the experiment to see the concentration-response curve was performed in triple. Inhibitors were diluted to find% inhibition values at 6 or more concentrations, and the results obtained were determined by nonlinear regression fitting using program Prism ^™ (GraphPad software, San Diego, Calif.) To obtain IC ₅₀ values.

The results of the VEGFR-2 tyrosine kinase inhibitory effect measured by Experimental Examples 1-1 and 1-2 are shown in Table 2 .

Example % Inhibition (20 μM) Example % Inhibition (20 μM) Example % Inhibition (20 μM) Tyrphostin a51 88 18 - 36 57 One - 19 - 37 68 2 - 20 - 38 55 3 13 21 80 39 61 4 49 22 - 40 69 5 - 23 30 41 69 6 81 24 43 42 60 7 50 25 29 43 67 8 69 26 - 44 61 9 52 27 49 45 56 10 25 28 72 46 75 11 26 29 60 47 56 12 27 30 54 48 32 13 32 31 40 49 - 14 58 32 48 50 - 15 52 33 41 51 - 16 39 34 55 52 - 17 41 35 30

As shown in Table 2 , the control Tyrphostin A51 showed a strong inhibitory effect of 88% at 20 μM against VEGFR-2 tyrosine kinase, Examples 6, 8, 21, 28, 29, 37, Compounds 39-44 and 46 showed a good inhibitory effect of at least 60% against VEGFR-2 tyrosine kinase at 20 μM concentration. In particular, the compounds of Examples 6, 21, 28 and 46 showed more than 70% inhibitory effect. Therefore, since the compounds of the present invention have an inhibitory effect on VEGFR-2 tyrosine kinase, it can be usefully used as an inhibitor of tumor cell growth and metastasis through inhibition of neovascularization.

The results of EGFR tyrosine kinase inhibitory effect measured by Experimental Examples 1-1 and 1-2 are shown in Table 3 .

Example % Inhibition (20 μM) Example % Inhibition (20 μM) Example % Inhibition (20 μM) Tyrphostin a51 93 18 42 36 - One 31 19 38 37 - 2 - 20 - 38 - 3 - 21 64 39 - 4 - 22 20 40 81 5 22 23 - 41 - 6 74 24 - 42 54 7 - 25 - 43 54 8 64 26 - 44 65 9 42 27 - 45 42 10 19 28 67 46 71 11 19 29 - 47 52 12 35 30 - 48 - 13 20 31 - 49 - 14 - 32 - 50 - 15 - 33 - 51 - 16 37 34 44 52 - 17 - 35 27

As shown in Table 3 , the control Tyrphostin A51 showed a strong inhibitory effect of 93% at 20 μM against VEGFR-2 tyrosine kinase and Examples 6, 8, 21, 28, 40, 44, and Compounds of 46 showed a good inhibitory effect of at least 60% against VEGFR-2 tyrosine kinase at 20 μM concentration. In particular, the compounds of Examples 6, 40 and 46 showed more than 70% inhibitory effect. Therefore, the compounds of the present invention have an inhibitory effect on EGFR tyrosine kinase, and thus can be usefully used as an inhibitor of tumor cell growth and metastasis through cancer cell growth inhibition.

Experimental Example 2 Measurement of In vitro Growth Inhibitory Effect on Cancer Cell Lines

In vitro cytotoxicity experiments were carried out to determine the effect of 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivatives of the present invention on inhibiting the growth of cancer cell lines.

In vitro cytotoxicity experiments on various cancer cell lines were carried out in the following manner in accordance with the guidelines of the National Cancer Institute (NCI) (Skehan, P. et. Al., J. Natl. Cancer Inst., 82). : 1107, 1990). Cell culture medium RPMI 1640 medium, fetal bovine serum (FBS) and trypsin were purchased from Gibco (Gibco, Grand Island, NY), NaHCO ₃ , amphotericin B and gentamicin were used by Sigma Chemical. Reagents such as sulforhodamine B (SRB), trisma base, trichloro acetic acid (TCA) and doxorubicin, which are reagents used for cytotoxicity measurement experiments, were also purchased from Sigma Chemical. T-25, T-75 culture vessels used for cell culture, 96-well plates and disposable vitreous used for other cell culture were all used by Falcon, Lincoln Park, NJ. Microplate plate reader (ELISA reader) for measuring the cytotoxicity was used E-max model of Molecular Devices (Molecular Devices, Sunnyvale, CA). The compound used in the experiment was dissolved in a small amount of DMSO solution and diluted as the experimental medium to the desired concentration for the test so that the final DMSO concentration was 0.5% or less. As a control material, Iressa, an EGFR tyrosine kinase inhibitor from Astra Geneca, was used. The cancer cell lines used in the experiment were all purchased from the American Type Culture Collection (ATCC) and are human-derived cancer cell lines, ovarian cancer SK-OV-3 (HER-2 overexpressing cell line) and epithelial carcinoma A431 (EGFR). Overexpressing cell lines) cells (Rusnak, DW et. Al., Molecular Cancer Therapeutics., 1: 85-94, 2001; Konecny, Gottfried E. et. Al., Cancer Res., 66: 1630-1639, 2006 ). Culture medium was incubated in 37%, 5% CO ₂ incubation using RPMI 1640 medium added with 5% FBS, each passage was maintained every 3 to 4 days.

Cells were aliquoted into 96 well flat-bottom microplates and incubated for 24 hours to allow cells to adhere to the bottom side. After attaching the cells to the bottom surface, the culture solution was removed, and the culture solution containing each concentration of the test drug was added, and then cultured for 72 hours. After incubation with the compound, cytotoxicity was measured using SRB, a protein staining reagent. That is, after incubation with the drug, the culture solution was removed, treated with cold TCA solution in each well, and left at 4 ° C. for 1 hour to fix cells. Thereafter, the TCA was removed and dried at room temperature, and then stained with 1% acetic acid solution in which 0.4% SRB was dissolved and left at room temperature for 30 minutes to stain the cells. Extra SRB that did not bind to cells was removed by washing with 1% acetic acid solution, and SRB was eluted by adding 10 mM Trisma base-unbuffered solution of pH 10.3-10.5 to the stained cells. The absorbance of each well was measured at 520 nm using a microplate reader, and Tz was determined from the OD values of the wells (C) without drugs, the wells with each drug (T), and the wells (Tz) when the drug was first applied. Cytotoxicity of drugs with the formula [(T-Tz) / (C-Tz)] × 100 for ≤T and [(T-Tz) / (Tz)] × 100 for Tz> T Was calculated. The calculation results are shown in Table 4 .

 Example IC ₅₀ (μM) SK-OV-3 A431 5 14.2 15.3 7 9.04 6.21 16 6.12 5.32 17 5.30 4.04 46 0.82 1.10

The compounds of Examples 16 and 17 exhibited moderate inhibitory effects with IC ₅₀ values of about 5 μM against SK-OV-3, the HER-2 overexpressing cell line, and A431, the overexpressing cell line E431, and the compound of Example 46. Showed an IC ₅₀ value of 0.82 and 1.10 μM for HER-2 overexpressing cell lines SK-OV-3 and A431, respectively, and showed an excellent inhibitory effect on growth factor overexpressing cancer cell lines. Therefore, the compounds of the present invention may be useful as anticancer agents because they exhibit cytotoxicity against growth factor overexpressing tumor cells and inhibit cancer cell growth.

On the other hand, the compound according to the present invention can be formulated in various forms according to the purpose. The following are some examples of formulation methods containing the compound according to the present invention as an active ingredient, but the present invention is not limited thereto.

Preparation Example 1 Tablet (Direct Pressurization)

5.0 mg of the derivative compound of the present invention

Lactose 14.1 mg

Crospovidone USNF 0.8 mg

0.1 mg magnesium stearate

After sifting the derivative compound of the present invention, lactose, crospovidone USNF and magnesium stearate were mixed and pressed to prepare a tablet.

Preparation Example 2 Tablet (Wet Assembly)

5.0 mg of the derivative compound of the present invention

Lactose 16.0 mg

Starch 4.0mg

Polysorbate 80 0.3 mg

Colloidal silicon dioxide 2.7 mg

Magnesium Stearate 2.0mg

Distilled water

After sifting the derivative compound of the present invention, lactose and starch were mixed. After dissolving Polysorbate 80 in distilled water, an appropriate amount of this solution was added and then atomized. After drying, the fine particles were sieved and mixed with colloidal silicon dioxide and magnesium stearate. The granules were pressed to produce tablets.

Preparation Example 3 Powder and Capsule

5.0 mg of the derivative compound of the present invention

Lactose 14.8 mg

Polyvinyl pyrrolidone 10.0 mg

Magnesium Stearate 0.2mg

After sifting the derivative compound of the present invention, it was mixed with lactose, polyvinyl pyrrolidone and magnesium stearate. The mixture was compressed into tablets according to a conventional capsule preparation method and filled into gelatin capsules to prepare gelatin capsules.

<Example 4> Injection

100 mg of the derivative compound of the present invention

Mannitol 180 mg

_{Na 2 HPO 4 · 12H 2 O} 26 ㎎

Distilled water 2974 mg

The derivative compound of the present invention was dissolved in distilled water together with mannitol and Na ₂ HPO ₄ .12H ₂ O, sterilized by adjusting the pH to about 7.5, and then injections were prepared according to a conventional method.

As described above, the 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative of the present invention is tyrosine of vascular endothelial growth factor (VEGF) or epidermal growth factor (EGF) receptor By showing an inhibitory effect on kinases and a cytotoxic effect on cell growth factor overexpressing cancer cell lines, the pharmaceutical composition of the present invention inhibits neovascularization and cancer cell growth signaling, thereby inhibiting tumor cell growth and metastasis. Can be usefully used as an anticancer agent.

Claims (14)

In the 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative represented by Formula 1 or a pharmaceutically acceptable salt thereof, [Formula 1]

The derivative, 2- (3-chloro-4-methoxyanilino) -4- (3- (2- (piperidin-1-yl) ethoxy) phenyl) thiazole; 2- (3-chloro-4-methoxyanilino) -4- (3- (2- (morpholin-1-yl) ethoxy) phenyl) thiazole; And 2, which is a compound selected from the group consisting of 2- (3-chloro-4-methoxyanilino) -4- (4-bromo-3- (piperidin-1-yl) phenyl) thiazole -(3-chloro-4-methoxy) anilino-4-arylthiazole derivative or a pharmaceutically acceptable salt thereof. delete delete delete delete delete delete delete delete delete delete A pharmaceutical for preventing and treating cancer containing 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative represented by Formula 1 as an active ingredient or a pharmaceutically acceptable salt thereof Composition. The method of claim 12, wherein the cancer is liver cancer, stomach cancer, breast cancer, colon cancer, bone cancer, pancreatic cancer, head or neck cancer, uterine cancer, ovarian cancer, rectal cancer, esophageal cancer, small intestine cancer, anal muscle cancer, colon cancer, fallopian tube carcinoma, endometrial carcinoma Cervical carcinoma, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, prostate cancer, bladder cancer, kidney cancer, ureter cancer, renal cell carcinoma, renal pelvic carcinoma and central nervous system tumor Pharmaceutical composition for preventing and treating cancer. Pharmaceutical for inhibiting angiogenesis, comprising as an active ingredient a 2- (3-chloro-4-methoxy) anilino-4-arylthiazole derivative of claim 1 or a pharmaceutically acceptable salt thereof Composition.