THIENO [3,2-d] PYRIMIDINE DERIVATIVES HAVING

INHIBITORY ACTIVITY ON PROTEIN KINASES

FIELD OF THE INVENTION

The present invention relates to novel thieno[3,2-d]pyrimidine derivatives having inhibitory activity on protein kinases and a pharmaceutical composition for preventing or treating abnormal cell growth diseases. BACKGROUND OF THE INVENTION

Protein kinases, a catalyst enzyme for phosphorylation of hydroxyl groups in tyrosine, serine and threonine moieties of protein, play an important role in a growth factor signal transduction inducing growth, differentiation and proliferation on cells.

In order to maintain homeostasis in body, a signal transduction system in body should keep a balance between on and off. However, a mutation or an overexpression of particular protein kinases collapses the signal transduction system in a normal cell, e.g., by a continuous signal transduction in body, to induce various diseases such as cancer, inflammation, metabolic disease, and brain disease. Human protein kinases are estimated to exist in 518 species, about 1.7% of human whole genes (Manning et al., Science, 2002, 298, 1912), and largely divided into tyrosine protein kinases (at least 90 species) and serine/threonine protein kinases. Tyrosine protein kinases may be divided into receptor tyrosine kinases which are classified into 20 subtypes of 58 species and cytoplasm/non- receptors which are classified into 10 subtypes of 32 species. Receptor tyrosine kinases have domains on the cell surface for reception of growth factors, and active sites in the cytoplasm for phosphorylation of tyrosine moieties. When a growth factor binds to the growth factors receptor site on the cell surface of the receptor tyrosine kinase, the receptor tyrosine kinase forms a polymer and the tyrosine moieties of cytoplasm are autophosphorylated. Then, the signal transduction progresses inside nuclear by sequential phosphorylation of subfamily proteins, and finally, transcription factors inducing cancer are overexpressed.

A chromosome translocation between Bcr (breakpoint cluster region) genes located in chromosome 22 and Abl (V-abl Abelson murine leukemia viral oncogene homolog) genes located in chromosome 9 by chromosomal instability, generates oncogenes of Bcr- Abl genes. The Bcr-Abl chromosome translocation is called Philadelphia chromosome (Nowell and Hungerford, J. Natl. Cancer Inst., 1960;25:85). In the Bcr-Abl gene, the Bcr part has oligomerization domains and the Abl part has tyrosine kinase domains. The size of the Bcr-Abl gene is determined by the cut position of the Bcr gene, and 3 subtypes (190, 210, 230 kDa) of Bcr-Abl genes have been reported. The Bcr-Abl gene is a leukemia- inducing factor, particularly p210-Bcr-Abl is a direct tumor factor inducing chronic myeloid leukemia (CML). The correlation between p210-Bcr-Abl and the CML induction is very high (>98%). Novartis developed Gleevec (imatinib mesylate) which can selectively inhibit Bcr-Abl, i.e., by inhibiting tyrosine kinases of Abl, and released in 2002. Gleevec, the first targeted antitumor agent, is being widely used as an initial standard therapy for treating CML due to its characteristic property and excellent stability. However, inactivation of Gleevec by the acquired resistance became a problem. The most important factor among various factors inducing the acquired resistance is point- mutation generated in the Abl kinase domains. There were attempts to overcome the acquired Gleevec-resistance by inhibiting such point-mutant species. Nilotinib and dasatinib, which are recently available, effectively inhibit many point-mutant species generated by the acquired Gleevec-resistance in Abl kinase domains. Among many point-mutant species generated by the acquired Gleevec-resistance in Abl kinase domains, the most important is T315I-Bcr-Abl mutant species wherein threonine 315 as a gatekeeper of Abl kinase is substituted with isoleucine. However, nilotinib and dasatinib are unable to inhibit T315I-Bcr-Abl mutant species. Accordingly, there are many attempts to develop a medicine inhibiting T315I-Bcr-Abl mutant species.

A vascular endothelial growth factors receptor (VEGFR) of a receptor tyrosine kinase (RTK) is an important modulator for angiogenesis. It is involved in generations of vascular and lymphatic vessel and in homeostasis as well as has an important effect on nerve cells. Vascular endothelial growth factors (VEGF) are predominantly produced by vascular endothelial, hematopoietic and stromal cells in response to hypoxia and upon stimulation with growth factors such as TGFs, interleukins or PDGF. VEGFs bind to VEGF receptor (VEGFR)- 1, -2, and -3, and each VEGF isoform binds to a particular subset of these receptors giving rise to the formation of receptor homo- and heterodimers that activate discrete signaling pathways. Signal specificity of VEGF receptors is further modulated upon recruitment of coreceptors, such as neuropilins, heparan sulfate, integrins or cadherins.

The biological functions of VEGFs are mediated upon binding to type III RTKs, VEGFR- 1 (Flt-1), VEGFR-2 (KDR/Flk-1) and VEGFR-3 (Flt-4). VEGFRs are closely related to Fms, Kit and PDGFRs, VEGF bind to each specific receptor, VEGF-A binds to VEGFR-1 and -2 and to receptor heterodimers, while VEGF-C and -D bind VEGFR-2 and -3. P1GF and VEGF-B exclusively bind VEGFR-1 and VEGF-E interacts only with VEGFR-2. VEGF-F variants interact with either VEGFR-1 or -2. VEGF-A, -B and P1GF are predominantly required for blood vessel formation, while VEGF-C and -D are essential for the formation of lymphatic vessels. Angiogenesis provides tumors with nutrients, oxygen, and path for cancer cell spread so as to be essential for proliferation and spread. Angiogenesis in normal body is balanced by co-regulation of angiogenic stimulators and angiogenic suppressors, while in off-balanced cancer cells VEGFR is activated by growth factors (VEGF) which have a great effect on vascular endothelial cells. Various inhibitors of VEGF receptor tyrosine kinases using low molecular synthetic materials are being developed, most of which are able to be used for solid tumors and to inhibit angiogenesis activated only in cancer cells and have an excellent medicinal effect with relatively low side effects.

Tie2, a kind of receptor tyrosine kinase, is deeply concerned with angiogenesis and vasculature. The domain structure of Tie2 is conserved in all vertebrates very well (Lyons et al., Isolation of the zebrafish homologues for the tie-1 and tie-2 endothelium- specific receptor tyrosine kinases., Dev Dyn., 1998; 212:133-140). Tie2 ligands are angiopoietins (Ang). Ang2 does not induce Tie2 autophosphorylation and disturbs Tie2 activation induced by Angl. In endothelial cells, Tie2 activation by Ang2 induces PI3K- Akt activation (Jones et al., Identification of Tek/Tie2 binding partners. Binding to a multifunctional docking site mediates cell survival and migration., J Biol Chem., 1999; 274:3089630905). In mitogen-activated protein kinases (MAPK) signal transduction path as main signal transduction system of Tie2, adapter protein GRB2 and protein tyrosine phosphatase SHP2 play an important role in dimerization process by autophosphorylation of Tie2 receptor tyrosine kinases. Ang/Tie2 and vascular endothelial growth factors (VEGF) signal transduction path perform an important function in angiogenesis of cancer cells. Tie2 is manifested in vascular endothelial, particularly in the infiltration area of cancer cells. Tie2 overexpressions are found in breast cancer (Peters et al., Expression of Tie2/Tek in breast tumour vasculature provides a new marker for evaluation of tumour angiogenesis. Br J Cancer, 1998; 77:5156) and also in uterine cancer, liver cancer, and brain cancer.

RET (rearranged during transfection), a kind of receptor tyrosine kinases is expressed mostly in nerve cells and endocrine system. N-terminal intermolecular domains of RET consist of 4 N-cadherin-like repeats, calcium-binding sites, 9 N- glycosylation sites, and cysteine-rich regions (Aiaksinen et al., Nat. Rev. Neurosci., 2002;3:383). The cytoplasm area of RET has at least 12 tyrosine autophosphorylation sites (Liu, J., Biol. Chem., 1996;271 :5309). For example, RET9 variants have 16 autophosphorylation sites in the kinase domain. When a GFL/GFR-alpha complex binds to the intermolecular domain of RET, RET is autophosphorylated and activated (Aiaksinen et al., Nat. Rev. Neurosci., 2002;3:383). GFL, GNDF (glial-derived neurotropic factor- family ligands, consists of GNDF, artemin, neurturin, and persephin. GFR-alpha having 4 subtypes of GFR-alpha 1-4 is known as glycosylphosphatidylinositol-anchored co- receptor. RET plays an important role in parasympathetic, enteric nervous systems, and kidney generation of mouse (Pachnis et al., Development, 1993; 119:1005). RET defunctionalization by germline mutation induce Hirschsprung's disease which is identified as a congenitial aganglionosis of distal intestines (Manie et al., Trends Genet., 2001; 17:580). However, mutations promoting the function of RET induce MEN2A (multiple endocrine neoplasia type 2A), MEN2B, and familial medullary thyroid carcinoma (FMTC). In particular, RET is proved to be a promising molecular target for development of a medicine of thyroid cancer (Cote and Gagel, N. Engl. J. Med., 2003 ;349: 1566).

SUMMARY OF THE INVENTION

Accordingly, it is an object of the present invention to provide novel thieno[3,2- djpyrimidine derivatives having inhibitory activity on protein kinases.

It is another object of the present invention to provide a pharmaceutical composition for preventing or treating abnormal cell growth diseases.

In accordance with an aspect of the present invention, there is provided a thieno[3,2-d]pyrimidine derivative of formula (I), or a pharmaceutically acceptable salt, hydrate and solvate thereof:

Y is -CT^CR ¹ -, -CC-, or -C(=0)NR'-;

L is -C(=0)NR ² -, -NR ² C(=0)-, or -NR ² C(=0)NR ² -;

R ¹ and R ² are each independently H, C^alkyl, or C _3-8 cycloalkyl;

R is H, halogen, methyl, or methoxy;

E is Cs-naryl or C ₂ -i ₃ heteroaryl, which is unsubstituted or substituted by 1 to 3 substituents selected from the group consisting of halogen, -N0 ₂ , -CN, -NH ₂ , -OH, -CF ₃ , C _]-6 alkyl, hydroxy-C _1-6 alkyl, C _3-g cycloalkyl, -(CH ₂ ) _n -C _1-6 alkylamino, -(CH ₂ ) _n -diCi. ₆ alkylamino, -(CH ₂ )„Ci. ₆ alkoxy, -(CH ₂ ) _n -OS(=0) ₂ -C _1-6 alkyl, -(CH ₂ ) _n -C _3-14 aryl, -(CH ₂ )„-C _{2- 13} heteroaryl and -(CH ₂ ) _n -C _2-13 heterocycloalkyl, wherein n is an integer number of 0 to 3, and the aryl, heteroaryl and heterocycloalkyl are each independently unsubstituted or substituted by a substituent selected from the group consisting of hydroxy-Ci. ₆ alkyl, halogen, diCi-ealkylamino, and Ci _-6 alkoxy; and

Z is H, -C(=0)R ³ , Ci _-6 alkyl, hydroxyQ-ealkyl, C _3-8 cycloalkyl, C _2-7 heterocycloalkyl, C _3-14 aryl, or C _2- i ₃ heteroaryl, wherein the aryl, heteroaryl, and heterocycloalkyl are each independently unsubstituted or substituted by C _1-6 alkyl, Ci _-6 alkoxy, C _3-8 cycloalkyl, R ³ -C _{2- 7} heterocycloalkyl or C2 _-7 heterocycloalkyl, and R ³ is C^alkyl, C _3-6 cycloalkyl, or phenyl.

In accordance with another aspect of the present invention, there is provided a pharmaceutical composition for preventing or treating abnormal cell growth diseases resulted from overexpression of a protein kinase, comprising the compound of formula (I), or a pharmaceutically acceptable salt, hydrate or solvate thereof.

The inventive thieno[3,2-d]pyrimidine derivatives, or a pharmaceutically acceptable salt, hydrate or solvate thereof has an excellent inhibitory activity on protein kinases, and a pharmaceutical composition comprising the same is effective in preventing or treating abnormal cell growth diseases.

DETAILED DESCRIPTION OF THE INVENTION

Hereinafter, the present invention is described in detail. In the inventive compound of formula (I), E preferably is phenyl, pyridinyl, quinolinyl, isoquinolinyl, indolyl, isoxazlyl, or pyrazolyl, which is unsubstituted or substituted by 1 to 3 substituents selected from the group consisting of halogen, -N0 ₂ , -CN, -NH ₂ , -OH, -CF ₃ , C _1-6 alkyl, C _3-8 cycloalkyl, -(CH ₂ ) _n -C _1-6 alkylamino, -(CH ₂ ) _n -diC _{1- 6} alkylamino, -(CH ₂ )„C _1-6 alkoxy, -(CH ₂ ) _n -OS(=0) ₂ -C _1-6 alkyl, -(CH ₂ ) _n -C _3-I4 aryl, -(CH ₂ )„-C _{2- 13} heteroaryl and -(CH ₂ ) _n -C _2- i ₃ heterocycloalkyl, wherein n is an integer number of 0 to 3. More preferably, the aryl is phenyl; heteroaryl is pyrrolyl or imidazolyl; and heterocycloalkyl is pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, or diazepanyl, wherein said aryl, heteroaryl and heterocycloalkyl are each independently unsubstituted or substituted by Ci^alkyl, halogen, diCi^alkylamino or C _1-6 alkoxy.

In the inventive compound of formula (I), Z is preferably H, methyl, ethyl, 2- hydroxyethyl, 2-morpholinoethyl, isopropyl, cyclopropyl, cyclopentyl, cyclopropylcarbonyl, benzoyl, phenyl, 4-methoxyphenyl, 4-(4-methylpiperidine-l- yl)phenyl, 4-(4-ethylpiperazin-l-yl)phenyl, pyridine-4-yl, pyridine-2-yl, 5-methylpyridine- 2-yl, or 6-methylpyridine-3-yl.

Preferable compound according to the present invention is a thieno[3,2- d]pyrimidine derivative selected from the group consisting of:

1) (E)-3-(2-(4-aminothieno[3,2-d]pyrimidine-7-yl)vinyl)-4-methy l-N-(3-

(trifluoromethyl)phenyl)benzamide;

2) (E)-3 -(2-(4-aminothieno [3 ,2-d]pyrimidine-7-yl)vinyl)-N-(4-((4-ethylpiperazine- 1 - yl)methyl)-3-(trifluoromethyl)phenyl)-4-methylbenzamide;

3) (E)-3-(2-(4-aminolMeno[3,2-d]pyrimidine-7-yl)vinyl)-N-(3-(4- ethylpiperazine- 1 -yl)-

5-(trifluoromethyl)phenyl)-4-methylbenzamide;

4) (E)-N-(3-(2-(4-aminothieno[3,2-d]pyrimidine-7-yl)vinyl)-4-me thylphenyl)-3-(4- methyl- 1 H-imidazole- 1 -yl)-5-(trifluoromethyl)benzamide;

5) (E)-N-(3-(2-(4-aminothieno[3,2-d]pyrimidine-7-yl)vinyl)-4-me thylphenyl)-4-(l- methylpiperidine-4-yloxy)-3-(trifluoromethyl)benzamide;

6) (E)-N-(3-(2-(4-aminomieno[3,2-d]pyrimidine-7-yl)vinyl)-4-met hylphenyl)-3-

(trifluoromethyl)benzamide;

7) (E)-N-(3-(2-(4-aminotmeno[3,2-d]pyrimidine-7-yl)vinyl)-4-met hylphenyl)-4-((4- ethylpiperazin- 1 -yl)methyl)-3-(trifluoromethyl)benzamide;

8) 4-amino-N-(2-methyl-5-(3-(trifluoromethyl)phenylcarbamoyl)ph enyl)thieno[3,2- d]pyrimidine-7-carboxamide;

9) 4-amino-N-(5-(3-methoxyphenylcarbamoyl)-2-methylphenyl)thien o[3 ,2- d]pyrimidine-7-carboxamide;

10) 4-amino-N-(2-methyl-5-(3-(4-methyl- 1 H-imidazole- 1 -yl)-5-

(trifluoromemyl)phenylcarbamoyl)phenyl)thieno[3,2-d]pyrim idine-7-carboxamide;

11) 4-amino-N-(5-(4-((4-ethylpiperazin-l-yl)methyl)-3-

(trifluoromemyl)phenylcarbamoyl)-2-methylphenyl)thieno[3, 2-d]pyrimidine-7- carboxamide

12) 4-amino-N-(5-(3 ,5-dimethoxyphenylcarbamoyl)-2-methylphenyl)thieno [3 ,2- d]pyrimidine-7-carboxamide;

13) 4-(4-methoxyphenylamino)-N-(2-methyl-5-(3-

(trifluoromethyl)phenylcarbamoyl)phenyl)thieno[3,2-d]pyri midine-7-carboxamide;

14) N-(2-methyl-5-(3-(trifluoromethyl)phenylcarbamoyl)phenyl)-4- (3,4,5- trimethoxyphenylamino)tWeno[3,2-d]pyrimidine-7-carboxamide;

15) N-(2-methyl-5-(3-(trifluoromethyl)phenylcarbamoyl)phenyl)-4- (6-methylpyridine-

3-ylamino)thieno[3,2-d]pyrimidine-7-carboxamide;

16) 4-(4-(4-ethylpiperazin-l-yl)phenylamino)-N-(2-methyl-5-(3- (trifluoromethyl)phenylcarbamoyl)phenyl)thieno[3,2-d]pyrimid ine-7-carboxamide;

17) 4-(isopropylamino)-N-(2-methyl-5-(3-

(trifluoromethyl)phenylcarbamoyl)phenyl)tWeno[3,2-d]pyrim idine-7-carboxamide;

18) N-(2-methyl-5-(3-(trifluoromethyl)phenylcarbamoyl)phenyl)-4-

(methylamino)thieno[3,2-d]pyrimidine-7-carboxamide;

19) 4-(2-hydroxyethylamino)-N-(2-methyl-5-(3-

(trifluoromethyl)phenylcarbamoyl)phenyl)thieno[3,2-d]pyri midine-7-carboxamide;

20) N-(2-methyl-5-(3 -(trifluoromethyl)phenylcarbamoyl)phenyl)-4-(2-

21) 4-(cyclopropylamino)-N-(2-methyl-5-(3- ( fluoromethyl)phenylcarbamoyl)phenyl)tWeno[3,2-d]pyrimidine-7 -carboxamide;

22) 4-(cyclopropylamino)-N-(2-methyl-5-(3-(2-methyl-lH-imidazole -l-yl)-5-

(trifluoromethyl)phenylcarbamoyl)phenyl)thieno[3,2-d]pyri midine-7-carboxamide

23) 4-(3-(4-(cyclopropylamino)thieno[3,2-d]pyrimidine-7-carboxam ido)-4- methylbenzamido)-2-(trifluoromethyl)benzyl acetate;

24) 4-(cyclopropylamino)-N-(5-(4-(hydroxymethyl)-3-

(trifluoromethyl)phenylcarbamoyl)-2-methylphenyl)Meno[3,2 -d]pyrimidine-7- carboxamide;

25) 4-(3-(4-(cyclopropylamino)thieno[3,2-d]pyriniidine-7-carboxa mido)-4- methylbenzamido)-2-(trifluoromethyl)benzyl methanesulfonate;

26) 4-(cyclopropylamino)-N-(2-methyl-5-(4-((4-methyl- 1 H-imidazole- 1 -yl)methyl)-3- (trifluoromethyl)phenylcarbamoyl)phenyl)thieno[3,2-d]pyrimid ine-7-carboxamide;

27) 4-(cyclopropylamino)-N-(2-methyl-5-(3 -(4-methyl- 1 H-imidazole- 1 -yl)-5-

(tTifluoromemyl)phenylcarbamoyl)phenyl)tWeno[3,2-d]pyrimi dine-7-carboxamide;

28) N-(5-(3-bromo-5-(trifluoromethylcarbamoyl)phenyl)-2-methylph enyl)-4- (cyclopropylamino)tWeno[3,2-d]pyrimidine-7-carboxamide;

29) 4-(cyclopropylamino)-N-(2-methyl-5-(6-morpholinopyridine-3- ylcarbamoyl)phenyl)thieno[3,2-d]pyrimidine-7-carboxamide;

30) 4-(cyclopropylamino)-N-(5-(6-(4-ethylpiperazine-l-yl)pyridin e-3-ylcarbanioyl)-2- methylphenyl)thieno[3,2-d]pyrimidine-7-carboxamide;

31) 4-(cyclopropylamino)-N-(5-(3-(2,4-dimethyl-l H-imidazole- 1 -yl)-5- ( fluoromethyl)phenylcarbamoyl)-2-methylphenyl)tMeno[3,2-d]pyr imi carboxamide;

32) 4-(cyclopropylamino)-N-(2-methyl-5-(4-((4-methyl- 1 ,4-diazepan- 1 -yl)methyl)-3- (trifluoromethyl)phenylcarbamoyl)phenyl)thieno[3,2-d]pyrimid ine-7-carboxam^ 33) (S)-4-(cyclopropylamino)-N-(5-(4-((3-(dimethylamino)pyiTolid ine-l-yl)methyl)-3- ( fluoromethyl)phenylcarbamoyl)-2-methylphenyl)thieno[3,2-d]py rimidine-7- carboxamide;

34) 4-(cyclopropylamino)-N-(5-(3-(4-hydroxymethyl)-lH-imidazole- l-yl)-5-

( fluoromethyl)phenylcarbamoyl)-2-methylphenyl)thieno[3,2-d]py rimidine-7- carboxamide;

35) 4-(cyclopropylamino)-N-(2-methyl-5-(4-(l-methylpiperidine-4- yloxy)-3-

(trifluoromethyl)phenylcarbamoyl)phenyl)t eno[3,2-d]pyrimidine-7-carboxam

36) 4-(cyclopropylamino)-N-(2-methoxy-5-(3-(4-methyl- 1 H-imidazole- 1 -yl)-5-

(trifluoromemyl)phenylcarbamoyl)phenyl)thieno[3,2-d]pyrim idine-7-carboxam 37) 4-(cyclopropylamino)-N-(2-methyl-5-(4-((4-methylpiperazine-l -yl)methyl)-3-

( fluoromethyl)phenylcarbamoyl)phenyl)tbieno[3,2-d]pyrimidine- 7-carboxamide; 38) 4-(cyclopropylamino)-N-(5 -(4-((4-ethylpiperazine- 1 -yl)methyl)-3 -

( fluoromethyl)phenylcarbamoyl)-2-methylphenyl)tMeno[3,2-d]pyr imidine-7- carboxamide;

39) 4-amino-N-(2-methyl-5-(3 -(trifluoromethyl)benzamido)phenyl)thieno [3 ,2- d]pyrimidine-7-carboxamide;

40) 4-(cyclopropylanlino)-N-(2-methyl-5-(4-mo holino-3-

( fluoromemyl)benzamido)phenyl)thieno[3,2-d]pyrimidine-7-carbo xamide;

41 ) 4-(cyclopropylamino)-N-(5-(3-(3-(dimethylamino)propylamino)- 5- (trifluoromemyl)benzamido)-2-methylphenyl)lhieno[3,2-d]pyrim idm

carboxamide;

42) 4-amino-N-(5-beiizamido-2-memylphenyl)tWeno[3,2-d]pyrimidine -7-carboxamide;

43) 4-amino-N-(5-(3,5-dimethoxybenzamido)-2-methylphenyl)thieno[ 3,2-d]pyrimidine-

7-carboxamide;

44) N-(5-benzamido-2-methylphenyl)-4-(5-methylpyridin-2-ylamino) thieno[3,2- d]pyrimidine-7-carboxamide;

45) N-(5-(3-(lH-pyrrol-l-yl)-5-(trifluoromethyl)benzamido)-2-met hylphenyl)-4- aminothieno[3,2-d]pyrimidine-7-carboxamide;

46) 4-amino-N-(5-(3-(dimethylamino)-5-(trifluoromethyl)benzamido )-2- methylphenyl)thieno[3 ,2-d]pyrimidine-7-carboxamide; ) N-(5-(3-(l H-imidazol- 1 -yl)-5-(trifluoromethyl)benzamido)-2-methylphenyl)-4- aminothieno[3,2-d]pyrimidine-7-carboxamide;

) 4-amino-N-(5-(3-fluoro-5-( fluoromemyl)benzamido)-2-memylphenyl)thieno[3,2- d]pyrimidine-7-carboxamide;

) 4-amino-N-(5-(4-((4-ethylpiperazine- 1 -yl)methyl)-3 -(trifluoromethyl)benzamido)-2- methylphenyl)thieno[3,2-d]pyrimidine-7-carboxamide;

) l-(4-(3-(4-aminotmeno[3,2-d]pyrimidine-7-carboxyamido)-4- methylphenylcarbamoyl)-2-(trifluoromethyl)benzyl)piperidine- 4-yl acetate;

) N-(2-methyl-5-(3-(trifluoromethyl)benzamido)phenyl)-4-(pheny lamino)thieno[3,2- d]pyrimidine-7-carboxamide;

) N-(2-methyl-5-(3-(trifluoromethyl)benzamido)phenyl)-4-(pyrid ine-4- ylamino)thieno[3,2-d]pyrimidine-7-carboxamide;

) N-(2-methyl-5-(3-(trifluoromethyl)benzamido)phenyl)-4-(pyrid ine-2- ylamino)thieno[3,2-d]pyrimidine-7-carboxamide;

) 4-amino-N-(5-(isoquinoline- 1 -carboxyamido)-2-methylphenyl)thieno [3,2- d]pyrimidine-7-carboxamide;

) 4-amino-N-(5-(isoquinoline-3-carboxyamido)-2-methylphenyl)th ieno[3,2- d]pyrimidine-7-carboxamide;

) 4-amino-N-(5-(4-methoxyquinoline-2-carboxyamido)-2-methylphe nyl)thieno[3,2- d]pyrimidine-7-carboxamide;

) N-(5-( 1 H-indole-2-carboxyamido)-2-methylphenyl)-4-aminothieno [3 ,2- d]pyrimidine-7-carboxamide;

) 4-amino-N-(2-memyl-5-(picolinamido)phenyl)thieno[3,2-d]pyrim idine-7- carboxamide;

) 4-amino-N-(2-methyl-5-(nicotinamido)phenyl)1hieno[3,2-d]pyri midine-7- carboxamide;

) 4-amino-N-(5-(isomcotinamido)-2-memylphenyl)tmeno[3,2-d]pyri midine-7- carboxamide;

) 4-amino-N-(2-methyl-5-(3-(2-methyl- 1 H-imidazole- 1 -yl)-5-

(trifluoromethyl)benzamido)phenyl)mieno[3,2-d]pyrimidine- 7-carboxamide;

) 4-amino-N-(5-(3-fluorophenylamido)-2-methylphenyl)thieno[3,2 -d]pyrimidine-7- carboxamide;

) 4-amino-N-(2-memyl-5-(2-(trifluoromethyl)benzamido)phenyl)th ieno[3,2- d]pyrimidine-7-carboxamide;

) (R)-4-amino-N-(5-(3-(3-(dimethylamino)pyrrolidine-l-yl)-5- (trifluoromethyl)benzamido)-2-methylphenyl)thieno [3 ,2-d]pyrimidine-7- carboxamide;

65) 4-amino-N-(5-(3-methoxybenzamido)-2-methylphenyl)thieno[3,2- d]pyrimidine-7- carboxamide;

66) 4-amino-N-(2-methyl-5-(4-(trifluorometliyl)berizamido)phenyl )thieno[3,2- d]pyrimidine-7-carboxamide;

67) N-(3-(4-aminothieno[3,2-d]pyrimidine-7-carboxyamido)-4-methy lphenyl)-5- cyclopropylisoxazol-3-carboxamide;

68) 4-amino-N-(5-( 1 -(4-fluorobenzyl)-3 -methyl- 1 H-pyrazole-5-carboxyamido)-2- methylphenyl)thieno[3,2-d]pyrimidine-7-carboxamide;

69) N-(2-methyl-5-(3-(Mfluoromethyl)benzam

d]pyrimidine-7-carboxamide;

70) 4-(cycloamino)-N-(2-methyl-5-(3-(trifluoromethyl)benzamido)p henyl)thieno[3,2- d]pyrimidine-7-carboxamide;

71) 4-(cyclopentylamino)-N-(2-methyl-5-(3-

(trifluorome1hyl)benzamido)phenyl)tmeno[3,2-d]pyrimidine- 7-carb

72) N-(2-methyl-5-(3-( fluoromethyl)benzaniido)phenyl)-4-(6-methylpyridine-3- ylamino)thieno [3 ,2-d]pyrimidine-7-carboxamide;

73) 4-(4-(4-ethylpiperazine-l-yl)phenylamino)-N-(2-methyl-5-(3-

(1rifluoromethyl)benzamido)phenyl)t^

74) 4-(cyclopropylamino)-N-(2-methyl-5-(3 -(4-methyl- 1 H-imidazole- 1 -yl)-5-

( fluoromethyl)beiizaimdo)phenyl)tWeno[3,2-d]pyrimidine-7-carb oxamide;

75) 4-(cyclopropylamino)-N-(2-methyl-5-(4-(4-methylpiperazin- 1 -yl)-3-

(trifluoromethyl)benzamido)phenyl)thieno[3,2-d]pyrimidine -7-carboxam

76) (S)-4-(cyclopropylamino)-N-(5-(4-((3-(dimethylamino)pyiTolid in- 1 -yl)methyl)-3-

(1xifluoromethyl)benzoamido)-2-memylphenyl)thieno[3,2-d]p yrimidine^ carboxamide;

77) 4-(cyclopropylamino)-N-(2-methyl-5-(4-((4-methyl- 1 H-imidazole- 1 -yl)methyl)-3 -

( fluoromemyl)benzoamido)phenyl)thieno[3,2-d]pyrimidine-7-carb oxamide;

78) N-(2-methyl-5-(3-(4-methyl-lH-imidazole-l-yl)-5-

(trifluoromethyl)benzoamido)phenyl)-4-(methylamino)thieno [3,2-d]pyrimidine-7- carboxamide;

79) 4-(ethylamino)-N-(2-methyl-5-(3-(4-methyl-lH-imidazole-l-yl) -5-

(trifluoromethyl)benzoamido)phenyl)thieno[3,2-d]pyrimidin e-7-carboxamide;

80) 4-(cyclopentylamino)-N-(2-methyl-5-(3-(4-methyl- 1 H-imidazole- 1 -yl)-5- (trifluoromethyl)benzoamido)phenyl)tMeno[3,2-d]pyrimidine-7- carboxam 81 ) N-(2-methyl-5-(3-(4-methyl- 1 H-imidazole- 1 -yl)-5-

(trifluoromethyl)benzoamido)phenyl)-4-(4-(4-methylpiperaz in- 1 - yl)phenylamino)thieno[3,2-d]pyrimidine-7-carboxamide;

82) 4-(cyclopropylamino)-N-(2-methyl-5-(4-(4-methyl- 1 H-imidazole- 1 -yl)-3-

( fluoromemyl)benzoamido)phenyl)mieno[3,2-d]pyrimidine-7-carbo xamide;

83) 4-(cyclopropylamino)-N-(2-methyl-5-(3 -(2-methyl- 1 H-imidazole- 1 -yl)-5-

(tTifluoromethyl)benzoamido)phenyl)thieno [3 ,2-d]pyrimidine-7-carboxamide;

84) (R)-4-(cyclopropylamino)-N-(5-(3-(3-(dimethylamino)pyrrolidi ne- 1 -yl)-5- (trifluoromemyl)benzoamido)-2-memylphenyl)thieno[3,2-d]pyrim idine-7- carboxamide;

85) 4-(cyclopropylamino)-N-(2-methyl-5-(3 -(4-methylpiperazin- 1 -yl)-5-

(trifluoromemyl)benzoamido)phenyl)tWeno[3,2-d]pyrimidine- 7-carboxamide;

86) 4-(cyclopropylamino)-N-(2-methyl-5-(3 -(morpholino-5- (trifluoromemyl)benzoamido)phenyl)tmeno[3,2-d]pyrimidine-7-c arboxamide;

87) 4-(cyclopropylamino)-N-(2-methyl-5-(4-((4-methyl- 1 ,4-diazepan- 1 -yl)methyl)-3-

(trifluoromemyl)benzoamido)phenyl)mieno[3,2-d]pyrimidine- 7-carboxamide;

88) 4-(cyclopropylamino)-N-(5-(4-(2,4-dimethyl- 1 H-imidazole- 1 -yl)-3-

( fluoromemyl)benzoamido)-2-melhylphenyl)thieno[3,2-d]pyrimidi ne-7- carboxamide;

89) 4-(cyclopropylamino)-N-(2-fluoro-5-(3-(4-methyl-l,4-diazepan e-l-yl)-5-

(tTifluoromemyl)benzoamido)phenyl)tWeno[3,2-d]pyrimidine- 7-carboxamide;

90) 4-(cyclopropylamino)-N-(5-(3-(2,4-dimethyl- 1 H-imidazole- 1 -yl)-5-

(trifluoromemyl)benzoamido)-2-methylphenyl)mieno[3,2-d]py rimidine-7- carboxamide;

91) N-(5-(3-(lH-imidazole-l-yl)-5-(trifluoromethyl)benzoamido)-2 -methylphenyl)-4-

(cyclopropylamino)thieno[3,2-d]pyrimidine-7-carboxamide;

92) (S)-4-(cyclopropylamino)-N-(5-(3-(3-(dimethylamino)pyrrolidi n-l-yl)-5-

(1rifluoromemyl)benzoamido)-2-memylphenyl)mieno[3,2-d]pyr imidine-7- carboxamide;

93) 4-(cyclopropylamino)-N-(2-methyl-5-(3-(4-methyl-l,4-diazepan -l-yl)-5-

(trifluoromemyl)benzoamido)phenyl)tMeno[3,2-d]pyrimidine- 7-carboxamide;

94) (R)-4-(cyclopropylamino)-N-(5-(4-(3-(dimethylamino)pyrrolidi ne- 1 -yl)-3-

(trifluoromethyl)benzoamido)-2-memylphenyl)thieno[3,2-d]p yrimidine-7- carboxamide; 95) (S)-4-(cyclopropylamino)-N-(5-(4-(3-(dim

(trifluoromethyl)benzoamido)-2-methylphenyl)thieno[3,2-d]pyr imidin

carboxamide;

96) N-(2-methyl-5-(3-(4-methyl-lH-imidazole-l-yl)-5- (trifluoromethyl)benzoamido)phenyl)-4-( 1 -methylpiperidine-4-ylamino)thieno[3 ,2- d]pyrimidine-7-carboxamide;

97) 4-(cyclopropylamino)-N-(5-(4-(diethylamino)-3-(1iifluorometh yl)benzoamid

methylphenyl)thieno[3,2-d]pyrimidine-7-carboxamide;

98) 4-(cyclopropylamino)-N-(2-methyl-5-(4-(l-methylpiperidine-4- ylamino)-3- (trifluoromethyl)benzoamido)phenyl)thieno[3,2-d]pyrimidine-7 -carboxam

99) N-(2-chloro-5-(3-(4-methyl-lH-imidazole-l-yl)-5-

( fluoromethyl)benzoamido)phenyl)-4-(cyclopropylamino)thieno[3 ,2- d]pyrimidine-7-carboxamide;

100) (R)-4-(cyclopropylamino)-N-(5-(4-(3-(dimethylamino)pyiTolidi ne-l-yl)-3- (trifluorome1hyl)benzoamido)-2-fluorophenyl)tWeno[3,2-d]pyri midine-7- carboxamide;

101) N-(2-chloro-5-(3-(2,4-dimethyl-lH-imidazole-l-yl)-5- ( fluoromethyl)benzoamido)phenyl)-4-(cyclopropylamino)thieno [3,2- d]pyrimidine-7-carboxamide;

102) 4-(cyclopropylamino)-N-(2-methyl-5-(3-(morpholinoamino)-5-

(trifluoromethyl)benzoamido)phenyl)tWeno[3,2-d]pyrimidine -7-carboxami 103) N-(2-methyl-5-(3-(4-methyl- 1 H-imidazole- 1 -yl)-5-

(trifluoromethyl)benzoamido)phenyl)-4-(phenylamino)thieno [3,2-d]pyrimidm^ carboxamide;

104) 4-(cyclopropylamino)-N-(2-fluoro-5 -(4-(4-methylpiperazine- 1 -yl)-3 - (trifluoromethyl)berizoamido)phenyl)tmeno[3,2-d]pyrirnidm

105) 4-(cyclopropylammo)-N-(5-(3-(2-(dimethylamino)ethylamino)-5- (trifluoromethyl)benzoamido)-2-memylphenyl)thieno[3,2-d]pyri midi

carboxamide;

106) (S)-4-(cyclopropylamino)-N-(5-(4-(3-(dimethylamino)pyrrolidi ne-l -yl)-3- ( fluoromemyl)beiizoamido)-2-fluorophenyl)thieno[3,2-d]pyrimid ine-7- carboxamide;

107) (S)-4-(cyclopropylamino)-N-(5-(3-(3-(dimethylamino)pyrrolidi ne-l-yl)-5- (trifluoromemyl)benzoamido)-2-fluorophenyl)trdeno[3,2-d]pyri midine-7- carboxamide; 108) 4-(cyclopropylamino)-N-(5-(3-(2,4-dimethyl- 1 H-imidazole- 1 -yl)-5- ( fluoromemyl)benzoamido)-2-fluorophenyl)thieno[3,2-d]pyrimidi ne-7- carboxamide;

109) 4-(cyclopropylamino)-N-(2-methyl-5-(3-(piperidine-l -yl)-5- (trifluoromemyl)benzoamido)phenyl)mieno[3,2-d]pyrimidine-7-c arboxamide;

110) 4-(cyclopropylamino)-N-(5-(3-(4-ethylpiperazin-l-yl)-5- ( fluoromemyl)benzoamido)-2-methylphenyl)mieno[3,2-d]pyrimidin e-7- carboxamide;

111) 4-(cyclopropylamino)-N-(2-methyl-5-(4-(l-methylpiperidine-4- yloxy)-3- (trifluoromemyl)benzoamido)phenyl)thieno[3,2-d]pyrimidine-7- carboxam

112) 4-(cyclopropylamino)-N-(2-methyl-5-(3 -(pyrrolidine- l-yl)-5- (trifluoromemyl)benzoamido)phenyl)thieno[3,2-d]pyrimidine-7- carboxam

113) (R)-4-(cyclopropylamino)-N-(2-memyl-5-(3-(2-methylpyrrolidin e-l-yl)-5- (trifluoromethyl)benzoamido)phenyl)mieno[3,2-d]pyrimidine-7- carboxam

114) 4-(cyclopropylan mo)-N-(5-(3-(diemylamino)-5-(trifluoromemyl)benzoamido)-2- memylphenyl)trueno[3,2-d]pyrimidine-7-carboxamide;

115) 4-(cyclopropylarmno)-N-(5-(3-(ethylamino)

memylphenyl)tMeno[3,2-d]pyrimidine-7-carboxamide;

116) 4-(cyclopropylamino)-N-(2-methyl-5-(3-(l-methylpiperidine-4- ylarnino)-5- (trifluoromemyl)beiizoamido)phenyl)tiiieno[3,2-d]pyrimidine- 7-c

117) 4-(cyclopropylamino)-N-(3 -(3 -(4-methyl-l H-imidazole- l-yl)-5- (trifluoromemyl)benzoamido)phenyl)tmeno[3,2-d]pyrimidine-7-c arboxamide;

118) 4-(cyclopropylamino)-N-(2-methoxy-5-(3-(4-methyl- 1 H-imidazole- 1 -yl)-5- (trifluoromemyl)berizoamido)phenyl)mieno[3,2-d]pyrimidine-7- carboxamide;

119) 4-(cyclopropylcarboxyamido)-N-(2-methyl-5-(3-(4-methyl- 1 H-imidazole- 1 -yl)-5- (trifluoromemyl)benzoamido)phenyl)tWeno[3,2-d]pyrimidine-7-c arboxamide;

120) 4-benzoamido-N-(2-methyl-5-(3-(4-methyl- 1 H-imidazole- 1 -yl)-5- (lrifluoromemyl)berizoamido)phenyl)thieno[3,2-d]pyrimidine-7 -carboxamide;

121) 4-(cyclopropylamino)-N-(2-fluoro-5-(3-(4-methyl- 1 H-imidazole- 1 -yl)-5- (trifluoromemyl)benzoamido)phenyl)tWeno[3,2-d]pyrimidine-7-c arboxamide;

122) 4-amino-N-(5-(3-(4-chloro-3-(trifluoromethyl)phenyl)ureido)- 2- memylphenyl)tbieno[3,2-d]pyrimidine-7-carboxamide;

123) 4-(cyclopropylamino)-N-(2-methyl-5-(3-(3-(4-methyl- 1 H-imidazole- 1 -yl)-5- (trifluoromemyl)phenyl)ureido)phenyl)mieno[3,2-d]pyrimidine- 7-carboxamide;

124) 3-((4-(cyclopropylamino)thieno[3,2-d]pyrimidine-7-yl)ethynyl )-4-methyl-N-(3-(4- methyl- 1 H-imidazole- 1 -yl)-5-(trifluoromethyl)phenyl)benzamide;

125) N-(3-((4-(cyclopropylamino)thieno[3,2-d]pyrimidine-7-yl)ethy nyl)-4- methylphenyl)-3-(4-methyl-lH-imidazole-l-yl)-5-(trifluoromet hyl)benzamide;

126) 3-((4-(cyclopropylamino)thieno[3,2-d]pyrimidine-7-yl)ethynyl )-4-methyl-N-(4-((4- methylpiperazine- 1 -yl)methyl)-3-(trifluoromethyl)phenyl)benzamide;

127) 3-((4-(cyclopropylamino)thieno[3,2-d]pyrimidine-7-yl)ethynyl )-N-(4-((4- ethylpiperazine- 1 -yl)methyI)-3 -(trifluoromethyl)phenyI)-4-methyIbenzamide;

128) 3-((4-(cyclopropylamino)tlueno[3,2-d]pyrimidine-7-yl)ethynyl )-4-methyl-N-(4-(l - methylpiperazine- 1 -yloxy)-3-(trifluoromethyl)phenyl)benzamide; and

a pharmaceutically acceptable salt, hydrate and solvate thereof.

In preparation of the inventive compound of formula (I), the starting material or intermediate may be the compounds of formulas (II) to (V):

wherein,

A is halogen, -OR ⁴ , -SR ⁴ , -S(=0)R ⁴ , -S(=0) ₂ R ⁴ , -NR ⁴ R ⁵ , or -NR ⁴ C(=0)R ⁵ ;

R ⁴ and R ⁵ are each independently H, Ci _-6 alkyl, C _3-8 cycloalkyl, -C(=0)R ⁶ , C _2- vheterocycloalkyl, C _3- i ₄ aryl, or C _2- i ₃ heteroaryl, wherein the aryl, heteroaryl and heterocycloalkyl are each independently unsubstituted or substituted by the substituent selected from the group consisting of Ci _-6 alkyl, C2-7cycloalkyl, C _1-6 alkoxy and C _{2- 7} heterocycloalkyl;

R ⁶ is H, C _1-6 alkyl or C _2-7 cycloalkyl.

X is -NH ₂ or -C(0)OH;

Y is -CHCR ⁷ -, -CC-, or -C(0)NR ⁷ -, wherein R ⁷ is H, C _1-6 alkyl or C _3-8 cycloalkyl; and

R is H, halogen, methyl or methoxy.

The inventive compound of formula (I), or a pharmaceutically acceptable salt, hydrate or solvate thereof exhibits an excellent inhibitory activity on protein kinases.

Accordingly, the present invention provides a pharmaceutical composition for preventing or treating abnormal cell growth diseases resulted from overexpression of a protein kinase, comprising the compound of formula (I), or a pharmaceutically acceptable salt, hydrate or solvate thereof as an active ingredient.

The protein kinase may comprise Bcr-Abl, FGFR, Fit, KDR, PDGFR, Fms, Kit,

Raf, Tie2, Src, and Ret. The pharmaceutical composition of the present invention has an excellent inhibitory activity on these kinases.

The abnormal cell growth disease may comprise stomach cancer, lung cancer, liver cancer, colorectal cancer, small intestinal cancer, pancreatic cancer, brain cancer, bone cancer, melanoma, breast cancer, sclerosing adenosis, uterine cancer, uterine cervical cancer, head and neck cancer, esophageal cancer, thyroid cancer, parathyroid cancer, renal cancer, sarcoma, prostate cancer, urethra cancer, bladder cancer, blood cancer, lymphoma, psoriasis and fibroadenoma.

For example, the blood cancer may be leukemia, multiple myeloma or myelodysplastic syndrome; and the lymphoma may be Hodgkin's disease or non-Hodgkin's lymphoma.

Accordingly, the present invention provides a method for preventing or treating abnormal cell growth diseases resulted from overexpression of a protein kinase, comprising administering the compound of formula (I), or a pharmaceutically acceptable salt, hydrate or solvate thereof to a subject requiring prevention or treatment of abnormal cell growth diseases resulted from overexpression of a protein kinase.

A proposed daily dose of the inventive compound for administration to a human (of approximately 70 kg body weight) may be in the range of 1 mg/day to 2,000 mg/day. The inventive compound may be administered in a single dose or in divided doses per day. It is understood that the daily dose should be determined in light of various relevant factors including the condition, age, body weight and sex of the subject to be treated, administration route, and disease severity; and, therefore, the dosage suggested above should not be construed to limit the scope of the present invention in anyway.

The present invention provides thieno[3,2-d]pyrimidine derivatives represented by formula (I) or a pharmaceutically acceptable salt thereof. The pharmaceutically acceptable salt should have a low toxicity in human and should not adversely affect the biological activity and physicochemical properties of the parent compound. The pharmaceutically acceptable salts comprise an acid addition salt of a pharmaceutically acceptable free acid and a base compound of formula (I); an alkali metal salt (e.g., sodium salt) and an alkaline earth metal salt (e.g., calcium salt); an organic base addition salt of an organic base and a carboxylic acid compound of formula (I); and an amino acid addition salt.

Preferable salt forms of inventive compound comprise a salt formed with an inorganic acid or an organic acid. Examples of the inorganic acid are hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, perchloric acid, and bromic acid. Examples of the organic acid are acetic acid, methanesulfonic acid, ethanesulfonic acid, p- toluenesulfonic acid, fumaric acid, malic acid, malonic acid, phthalic acid, succinic acid, lactic acid, citric acid, gluconic acid, tartaric acid, salicylic acid, maleic acid, oxalic acid, benzoic acid, aspartic acid, and glutamic acid. The organic base addition salt may be prepared using an organic base such as tris(hydroxymethyl)methylamine and dicyclohexylamine. The amino acid addition salt may be prepared using a natural amino acid such as alanine and glycine.

These salts may be prepared in a conventional manner. For example, the salts may be prepared by dissolving the compound of formula (I) in a water-miscible solvent such as methanol, ethanol, acetone and 1,4-dioxane; adding a free acid or free base thereto; and then crystallizing.

The pharmaceutical composition of the present invention comprising the bicyclic heteroaryl derivatives represented by formula (I), a pharmaceutically acceptable salt, solvate or hydrate thereof as an active ingredient, may further contain a conventional nontoxic pharmaceutically acceptable carrier, a reinforcing agent, or a diluent, to be formulated for parenteral administration or oral administration, e.g., in the form of a tablet, capsule, troches, solution or suspension.

The pharmaceutical composition of the present invention may comprise conventional additives such as a diluent, a sweetener, a binder, a solubilizing agent, a solubilizing co-agent, a wetting agent, an emulsifier, an isoosmotic agent, an absorbent, a disintegrator, an antioxidant, a preservative, a lubricant, a filler, and an aromatics. Examples of the additives are lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, silica, talc, stearic acid, sterin, magnesium stearate, magnesium aluminum silicate, starch, gelatin, tragacanth gum, alginic acid, sodium alginate, methyl cellulose, sodium carboxymethyl cellulose, agar, water, ethanol, polyethylene glycol, polyvinyl pyrrolidone, sodium chloride, calcium chloride, orange essence, strawberry essence, vanilla essence, etc.

Examples of the carrier employed in the injectable composition of the present invention are distilled water, a saline solution, a glucose solution, a glucose-like solution, alcohol, glycol ether (e.g., polyethylene glycol 400), oil, fatty acid, fatty acid ester, glyceride, a surfactant, a suspension agent and an emulsifier.

The following Examples are intended to further illustrate the present invention without limiting its scope.

The compound of Preparation Example 1 was prepared according to Reaction Scheme 1 below: Reaction Scheme 1

Preparation Example 1: 7-Bromo-4-(methylthio)thieno[3,2-d]pyrimidine Step 1 : Thieno[3,2-d]pyrimidine-4(3H)-one Methyl-3-aminothiophene-2-carboxylate (15g, 98.57mmol) (Matrix, Cat # 018289, CAS [22288-78-4]) was dissolved in formamide (50mL), and the mixture was stirred at 180°C for 5 hours. The reaction mixture was further stirred at room temperature for 2 hours and then filtered to obtain the title compound.

Ή NMR (400MHz, DMSO-d ₆ ) δ 12.48 (br, 1H), 8.18 (d, J = 5.1 Hz, 1H), 8.14 (s,

1H), 7.40 (d, J = 5.1 Hz, 1H).

Step 2: 7-Bromothieno[3,2-d]pyrimidine-4(3H)-one

Thieno[3,2-d]pyrimidine-4(3H)-one (4.9g) was dissolved in acetic acid (20mL) and then brome (5mL) was added thereto. In a sealed reactor, the reaction mixture was stirred at 120°C for 10 hours. The reaction mixture was cooled to room temperature and distilled under reduced pressure to remove acetic acid. The resulting mixture was added to ice water and the resulting solid was filtered and then dried to obtain the title compound without further purification.

Ή NMR (400MHz, DMSO-d ₆ ) δ 12.75 (brs, 1H), 8.36 (s, 1H), 8.24 (s, 1H).

Step 3: -Bromo-4-cUoromieno[3,2-d]pyrimidine

7-Bromothieno[3,2-d]pyrimidine-4(3H)-one (5.9g) was dissolved in POCl ₃ (20mL) and then stirred at 150°C for 3 hours. After cooling down to room temperature, the remaining POCI ₃ was concentrated and added to ice water to obtain a solid. The solid was washed with sat. NaHC0 ₃ solution and dried with N ₂ gas to obtain the title compound.

Ή NMR (400MHz, DMSO-d^) δ 9.16 (s, 1H), 8.79 (s, 1H).

Step 4: 7-Bromo-4-(methylthio)thieno[3,2-d]pyrimidine

7-Bromo-4-chlorothieno[3,2-d]pyrimidine (2g, 8.07mmol) was dissolved in THF (27mL) and then NaSMe (650mg, 9.28mmol) was added at 0°C thereto. After stirring for 15 hours, ice water was added thereto. The resulting solid was filtered and dried with N ₂ gas to obtain the title compound without further purification.

Ή NMR (400MHz, DMSO-d ₆ ) δ 9.09(s, 1H), 8.57(5, 1H), 2.76(5, 3H).

The compound of Example 1 was prepared according to Reaction Scheme 2 below:

Reaction Scheme 2

Example 1: (E)-3-(2-(4-ammothieno[3,2-d]pyrimidme-7-yl)vinyl)-4-methyl- N-(3- (trifluoromethyl)phenyl)benzamide

Step 1 : -Bromothieno[3,2-d]pyrimidine-4-amine

7-Bromo-4-chlorothieno[3,2-d]pyrimidine (5g, 20.17mmol) obtained in Step 3 of

Preparation Example 1 and 2.0M NH ₃ dissolved in isopropanol solution (50mL) were added to a sealed reaction vessel and stirred at 100°C for 12 hours. The reaction mixture was cooled to room temperature and filtered to obtain a solid. The solid was washed with water and dried with N ₂ gas to obtain the title compound (3.8g, 83%) without further purification. MS m z [M+l] 229.97, 231.98.

Step 2: 7-Vinylthieno[3,2-d]pyrimidine-4-arnine 7-Bromothieno[3,2-d]pyrimidine-4-amine (2.4g, 10.43mmol) was dissolved in dioxane (30mL) and then treated with N ₂ gas for 20 minutes. Pd(PPh ₃ )4 (723mg, 0.63mmol) and tributyl(vinyl)tin (3.35mL, 11.5mmol) were added thereto and stirred at 120°C for 7 hours. The reaction mixture was cooled to room temperature, an aqueous KF solution (10%, 30mL) was added thereto and stirred for 2 hours. The reaction mixture was filtered with Celite and washed with ethyl acetate. The aqueous layer was separated from the organic layer and extracted with ethyl acetate. The combined organic layer was dried with MgS0 ₄ and concentrated. The resulting mixture was purified by silica gel chromatography (DCM/MeOH = 95/5) to obtain the title compound having a yellow color.

1H NMR (400MHz, DMSO-de) δ 8.41 (s, 1H), 8.14 (s, 1H), 6.95 (dd, 1H), 6.36 (d, 1H), 5.38 (d, 1H).

Step 3: E)-emyl-3-(2-(4-aminothieno[3,2-d]pyrimidine-7-yl)vinyl)-4-m ethyl benzoate

7-Vinylthieno[3,2-d]pyrimidine-4-amine (2.5g, 14.12mmol), ethyl-3-iodo-4-methyl benzoate (4.1 g, 14.12mmol), Pd(0 Ac) ₂ ( 190mg, 0.85mmol), tri-(2-furyl)phosphine (295mg, 1.27mmol), and DIEA (4.7mL, 28.24mmol) were dissolved in DMF (47mL) and then stirred at 120°C for 18hr. The reaction mixture was filtered and extracted with ethyl acetate. The organic layer was washed with brine, dried with MgS0 ₄ , filtered and concentrated. The resulting mixture was purified by silica gel chromatography to obtain the title compound (3.3g, yield 69%).

Ή NMR (400MHz, DMSO-d ₆ ) δ 8.48 (s, 1H), 8.34 (s, 1H), 8.19 (s, 1H), 8.14 (d, 1H), 7.75 (d, 1H), 7.50 (s, 2H), 7.39 (d, 1H), 7.36 (d, 1H), 4.32 (q, 2H), 2.49 (s, 3H), 1.32 (t, 3H); MS m/z [M+l] 340.28. Step 4: (E)-3-(2-(4-aminothieno[3,2-d]pyrimidine-7-yl)vinyl)-4-methy lbenzoic acid

(E)-emyl-3-(2-(4-aminothieno[3,2-d]pyrimidine-7-yl)vinyl)-4- methyl benzoate (3.0g, 8.84mmol) was dissolved in a mixture of methanol (15mL) and THF (15mL) and LiOH (1.8g, 44.19mmol) dissolved in water (15mL) was added thereto. The reaction mixture was stirred at room temperature for 18 hours. The organic layer was concentrated and IN HC1 solution was added dropwise thereto to obtain a solid. The solid was filtered and dried with N ₂ gas to obtain the title compound (2.45g, 89% yield) without further purification.

MS m/z [M+l] 312.17.

Step 5 : (E)-3 -(2-(4-aminothieno[3 ,2-d]pyrimidine-7-yl)vinyl)-4-methyl-N-(3 -

(trifluoromethyl)phenyl)benzamide

(E)-3-(2-(4-aminothieno[3,2-d]pyrimidine-7-yl)vinyl)-4-methy lbenzoic acid (30mg, 0.096mmol) was dissolved in DMF (lmL), 3-(trifluoromethyl)benzeneamine (16mg, 0.096mmol), HATU (l lOmg, 0.289mmol) and TEA (67μΙ,, 0.48mmol) was added thereto and stirred at room temperature for 18 hours. The reaction mixture was diluted with ethyl acetate and washed with brine. The organic layer was dried with MgS0 ₄ , filtered and concentrated. The resulting mixture was purified by silica gel chromatography to obtain the title compound (36mg, 82% yield).

MS m/z [M+l] 455.20.

Example 2 : (E)-3-(2-(4-aminothieno [3,2-dJ pyrimidine-7-yl) vinyl)-N-(4-((4- ethylpiperazme-l-yl)methyl)-3-(trifluoromethyl)phenyl)^-meth ylbenzamide

The procedure of Step 5 of Example 1 was repeated except for using (E)-3-(2-(4- aminothieno[3,2-d]pyrimidine-7-yl)vinyl)-4-methylbenzoic acid and 4-((4-ethylpiperazine- l-yl)methyl)-3-(trifluoromethyl)aniline to obtain the title compound (see Table 1). Example 3: (E)-3-(2-(4-aminothieno [3,2-d] pyrimidine-7-yl)vinyl)-N-(3-(4- ethylpiperazine-l-yl)-5-(trifluoromethyl)phenyl)-4-methylben zamide

The procedure of Step 5 of Example 1 was repeated except for using (E)-3-(2-(4- aminothieno[3,2-d]pyrirnidine-7-yl)vinyl)-4-methylbenzoic acid and 3-(4-ethylpiperazine- l-yl)-5-(trifluoromethyl)aniline to obtain the title compound (see Table 1).

The compound of Example 4 was prepared according to Reaction Scheme 3 below:

Reaction Scheme 3

Example 4 : (E)-N-(3-(2-(4-aminothieno [3,2-d] pyrimidine-7-yl) vinyl)-4-methylphenyl)- 3-(4-methyl-lH-imidazole-l-yl)-5-(trifluoromethyl)benzamide

Step 1 : (E)-7-(2-methyl-5-mtrostyryl)thieno[3,2-d]pyrimidine-4-amine

The procedure of Step 3 of Example 1 was repeated except for using 2-iodo-l- methyl-4-nitrobenzene (TCI Laboratory Chemicals, Cat. # 10706, CAS [7745-92-8]) instead of ethyl-3-iodo-4-methyl benzoate to obtain the title compound.

1H NMR (400 MHz, DMSO-d ₆ )5 8.50 (s, 1H), 8.44 (d, 1H), 8.37 (s, 1H), 8.25 (d, 1H), 8.04 (d, 1H), 7.56(d, 1H), 7.52(d, 1H), 7.45(s, 2H), 2.58(s, 3H); MS m/z [M+l] 313.25.

Step 2: (E)-7-(5-amino-2-memylstyryl)thieno[3,2-d]pyrimidine-4-amine

(E)-7-(2-memyl-5-nitros1yryl)thieno[3,2-d]pyrimidine-4-amine (0.3g, 0.96mmol) was dissolved in ethyl acetate (5mL) and then SnCl ₂ -2H ₂ 0 (lg, 4.81mmol) and cone. HC1 (0.5mL) were added thereto, and the reaction mixture was stirred at room temperature for 18 hours. After the reaction was completed NH ₄ OH solution was added thereto to pH 5. Then, anhydrous Na ₂ C0 ₃ was added thereto to pH 7. The reaction mixture was filtered with Celite and washed with ethyl acetate several times. The filtrate was concentrated under reduced pressure to obtain the title compound (260mg, 84% yield) without further purification.

1H NMR (400MHz, DMSO-d^ 8.54(s, 1H), 8.05(s, 1H), 8.02(s, 1H), 7.59(d, 1H), 7.28(d, 2H), 7.14(d, 1H), 6.57(d, 2H), 2.98(m, 1H), 0.80(m, 2H), 0.65(m, 2H); MS m/z 309.32 [M+l].

Step 3 : (E)-N-(3-(2-(4-aminothieno[3,2-d]pyrimidine-7-yl)vinyl)-4-me thylphenyl)-3-(4- methyl- 1 H-imidazole- 1 -yl)-5-(trifluoromethyl)benzamide

(E)-7-(5-amino-2-methylstyryl)thieno[3,2-d]pyrimidine-4-amin e (1 OOmg,

0.354mmol) and 3-(4-methyl-lH-imidazole-l-yl)-5-(trifluoromethyl)benzoic acid (96mg, 0.354mmol) were dissolved in DMF (1.5mL), HATU (404mg, 1.063mmol) and TEA (0.248mL, 1.7728mmol) were added thereto and stirred at room temperature for 18 hours. The reaction mixture was diluted with ethyl acetate and washed with brine. The organic layer was dried with MgS0 ₄ , filtered and concentrated. The resulting mixture was purified by silica gel chromatography to obtain the title compound (142mg, 75% yield).

MS m/z [M+l] 535.15 . Example 5: (E)-N-(3-(2-(4-aminothieno [3,2-d] pyrimidine-7-yl)vinyl)-4-methylphenyl)- 4-(l-methyIpiperidine-4-yloxy)-3-(trifluoromethyl)benzamide

The procedure of Step 3 of Example 4 was repeated except for using 4-(l- methylpiperidine-4-yloxy)-3-(trifluoromethyl)benzoic acid instead of 3-(4-methyl-lH- imidazole- l-yl)-5-(trifluoromethyl)benzoic acid to obtain the title compound (see Table 1).

Example 6: (E)-N-(3-(2-(4-ainmothieno[3,2-d]pyriinidme-7-yl)vmyl)-4-met liylphenyl)-

3- (trifluoromethyl)benzamide (E)-7-(5-amino-2-memylstyryl)tmeno[3,2-d]pyrimidine-4-amine (lOOmg,

0.354mmol) was dissolved in anhydrous THF, TEA (O.lmL, 0.709mmol) and 3- (trifluoromethyl)benzoyl chloride (81mg, 0.390mmol) were added thereto at room temperature and stirred for 4 hours. The reaction mixture was diluted with ethyl acetate and washed with brine. The organic layer was dried with MgS0 ₄ , filtered and concentrated. The resulting mixture was purified by silica gel chromatography to obtain the title compound (138mg, 86% yield) (see Table 1).

Example 7 : (E)-N-(3-(2-(4-aminothieno [3,2-d] pyrimidine-7-yl) vinyl)-4-methylphenyl)-

4- ((4-ethylpiperazin-l-yl)methyl)-3-(trifluoromethyl)benzamide

The procedure of Step 3 of Example 4 was repeated except for using 4-((4- ethylpiperazine-l-yl)methyl)-3-(trifluoromethyl)benzoic acid instead of 3-(4-methyl-lH- imidazole-l-yl)-5-(trifluoromethyl)benzoic acid to obtain the title compound (see Table 1). The compound of Preparation Example 2 was prepared according to Reaction

Scheme 4 below: Reaction Scheme 4

Preparation Example 2: 4-(Methylthio)thieno[3,2-d|pyrimidine-7-carboxylic acid

Step 1: -(Methylthio)-7-vinylthieno[3,2-d]pyrimidine

The procedure of Step 2 of Example 1 was repeated except for using 7-bromo-4- (methylthio)thieno[3,2-d]pyrimidine (500mg, 1.92mmol) obtained in Step 4 of Preparation Example 1 to obtain the title compound.

Ή NMR (400MHz, DMSO-d ₆ ) δ 9.05 (s, 1H), 8.40 (s, 1H), 7.02 (dd, 1H), 6.45 (d, 1H), 5.49 (d, 1H), 2.74 (s, 3H).

Step 2: 4-(Methylthio)thieno[3,2-d]pyrimidine-7-carbaldehyde

4-(Methylthio)-7-vinylthieno[3,2-d]pyrimidine (350mg, 1.68mmol) was dissolved in CHCl ₃ /MeOH (2mL/2mL) and treated with ozone at 78°C for 30 minutes. When the starting material was disappeared, the reaction mixture was treated with N ₂ gas at the same temperature for 5 minutes, heated to room temperature, and dimethylsulfide (0.37mL, 5.04mmol) was added thereto. The reaction mixture was concentrated to obtain a yellow solid. The solid was filtered, washed with diethyl ether and dried with N ₂ gas to obtain the title compound without further purification.

1H NMR (400MHz, DMSO-d ₆ ) δ 10.30 (s, 1H), 9.21 (s, 1H), 9.13 (s, 1H), 2.77 (s Step 3: 4-(Methylmio)thieno[3,2-d]pyrimidine-7-carboxylic acid

NaH ₂ P0 ₄ -2H ₂ 0 (513mg, 3.29mmol) was dissolved in water (2mL) and then 4- (methylthio)thieno[3,2-d]pyrimidine-7-carbaldehyde (300mg, 1.68mmol) dissolved in acetone (3mL) and DMSO (3mL) was added thereto at 0°C. NaC10 ₂ (194mg, 2.15mmol) dissolved in water (2mL) was added at the same temperature to the reaction mixture, which was stirred for 2 hours. After the reaction was completed water (lOmL) was further added to the reaction mixture, which was stirred for 2 hours and filtered to obtain a solid. The solid was washed with water several times and dried with N ₂ gas to obtain the title compound as a white solid.

1H NMR (400MHz, DMSO-d ₆ ) δ 8.98 (s, 1H), 8.49 (s, 1H), 2.72 (s, 1H).

The compound of Example 8 was prepared according to Reaction Scheme 5 below:

Reaction Scheme 5

Example 8: 4-Amino-N-(2-methyl-5- (trifluoromethyl)phenylcarbamoyl)phenyl)thieno[3,2-d]pyrimid ine-7-carboxamide

Step 1: Ethyl 4-methyl-3-(4-(methylthio)thieno[3,2-d]pyrimidine-7-carboxam ido)benzoate

4-(Methylthio)thieno[3,2-d]pyrimidine-7-carboxylic acid (2g, 10.2mmol) obtained in Step 3 of Preparation Example 2 was dissolved in DMF (40mL), methyl 3-amino-4- methyl benzoate (1.2g, 7.3mmol), HATU (8.3g, 22mmol) and DIPEA (6.4mL, 36.6mmol) were added thereto and stirred at 45°C for 18 hours. The reaction mixture was diluted with ethyl acetate and washed with an aqueous sodium bicarbonate solution and brine, sequentially. The organic layer was dried with MgS0 ₄ , filtered and concentrated. The resulting mixture was purified by silica gel chromatography to obtain the title compound (2.5g).

MS m/z [M+l] 388.09. Step 2: Ethyl 4-methyl-3-(4-(methylsulfinyl)thieno[3,2-d]pyrimidine-7- carboxamido)benzoate

Ethyl 4-methyl-3 -(4-(methylthio)thieno[3 ,2-d]pyrimidine-7-carboxamido)benzoate (500mg, 1.29mmol) was dissolved in DCM (6mL) and m-CPBA (290mg, 1.67mmol) was added thereto at 0°C. The reaction mixture was stirred at room temperature for 2 hours, diluted with DCM (30mL) and washed with sat. NaHC0 ₃ solution. The organic layer was dried with anhydrous MgS0 ₄ , filtered with Celite and concentrated to obtain the title compound without further purification.

MS m/z [M+l] 404.07.

Step 3: Ethyl 3-(4-aminothieno[3,2-d]pyrimidine-7-carboxamido)-4-methyl benzoate

The procedure of Step 1 of Example 1 was repeated except for using the compound obtained in Step 2 of Example 8 to obtain the title compound.

1H NMR (400 MHz, DMSO-d ₆ ) δ 11.89 (s, IH), 8.98 (s, IH), 8.96 (s, IH), 8.59 (s, IH), 7.95 (s, 2H), 7.66 (d, IH), 7.44 (d, IH), 4.32 (q, 2H), 2.54 (s, 3H), 1.33 (t, 3H); MS m/z [M+l] 356.9.

Step 4: 3-(4-Aminothieno[3,2-d]pyrimidine-7-carboxamido)-4-methylben zoic acid

The procedure of Step 4 of Example 1 was repeated except for using the compound obtained in Step 3 of Example 8 to obtain the title compound.

MS m/z [M+l] 329.1.

Step 5: 4-Ammo-N-(2-memyl-5-(3-(trifluoromemyl)phenylcarbamoyl)pheny l)thieno[3,2- d]pyrimidine-7-carboxamide

The procedure of Step 5 of Example 1 was repeated except for using the compound obtained in Step 4 of Example 8 to obtain the title compound.

1H NMR (300 MHz, DMSO-d ₆ )511.80 (s, IH), 10.47 (s, IH), 8.89 (s, IH), 8.80 (d, IH, J = 1.6 Hz), 8.52 (s, IH), 8.20-8.17 (m, IH), 8.00-7.97 (m, 2H), 7.95-8.95 (bs, 2H), 7.63-7.60 (m, IH), 7.52 (t, IH, J = 8.0 Hz), 7.40-7.36 (m, 2H), 2.51 (s, 3H).

Example 9: 4-Amino-N-(5-(3-methoxyphenylcarbamoyl)-2-methylphenyl)thien o[3,2- d] pyrimidine-7-car boxamide

The procedure of Step 5 of Example 1 was repeated except for using 3-(4- aminothieno[3,2-d]pyrimidine-7-carboxamido)-4-methylbenzoic acid obtained in Step 4 of Example 8 and 3-methoxyaniline to obtain the title compound (see Table 1).

Example 10: 4-Aitimo-N-(2-methyl-5-(3-(4-methyl-lH-imidazoIe-l-yl)-5- (trifluoromethyl)phenylcarbamoyl)phenyl)thieno [3,2-d] pyrimidine-7-carboxamide

The procedure of Step 5 of Example 1 was repeated except for using 3-(4- aminothieno[3,2-d]pyrimidine-7-carboxamido)-4-methylbenzoic acid obtained in Step 4 of Example 8 and 3-(4-methyl-lH-imidazole-l-yl)-5-(trifluoromethyl)aniline to obtain the title compound (see Table 1).

Example 11: 4-Amino-N-(5-(4-((4-ethylpiperazin-l-yl)methyl)-3-

(trifluoromethyl)phenylcarbamoyl)-2-methylphenyl)thieno[3 ,2-d]pyrimidine-7- carboxamide

The procedure of Step 5 of Example 1 was repeated except for using 3-(4- aminothieno[3,2-d]pyrimidine-7-carboxamido)-4-methylbenzoic acid obtained in Step 4 of Example 8 and 4-((4-emylpiperazin-l-yl)methyl)-3-(trifluoromethyl)benzenam ine to obtain the title compound (see Table 1).

Example 12: 4-Amino-N-(5-(3,5-dimethoxyphenylcarbamoyl)-2- methylphenyl)thieno[3,2-d]pyrimidine-7-carboxamide

The procedure of Step 5 of Example 1 was repeated except for using 3 -(4- aminothieno[3,2-d]pyrimidine-7-carboxamido)-4-methylbenzoic acid obtained in Step 4 of Example 8 and 3,5-dimethoxyaniline to obtain the title compound (see Table 1).

The compound of Example 13 was prepared according to Reaction Scheme 6 below: Reaction Scheme 6

Example 13: 4-(4-Methoxyphenylamino)-N-(2-methyl-5-(3- (Mfluoromethyl)phenylcarbamoyl)phenyl)thieno[3,2-d]pyrimidme -7-carboxamide

Step 1 : N-(2-methyl-5-(3-(trifluoromethyl)phenylcarbamoyl)phenyl)-4-

(memylthio)thieno[3,2-d]pyrimidine-7-carboxamide

The procedure of Step 1 of Example 8 was repeated except for using the compound obtained in Preparation Example 2 and 3-amino-4-methyl-N-(3- (trifluoromethyl)phenyl)benzamide to obtain the title compound.

MS m/z [M+l] 503.09. Step 2: N-(2-methyl-5-(3-(trifluoromethyl)phenylcarbamoyl)phenyl)-4-

(memyl xamide

The procedure of Step 2 of Example 8 was repeated except for using the compound obtained in Step 1 of Example 13 to obtain the title compound.

MS m/z [M+l] 519.08. Step 3: 4-(4-Methoxyphenylamino)-N-(2-methyl-5-(3-

(trifluorornethyl)phenylcarbamoyl)phenyl)thieno [ 3 ,2-d]pyrimidine-7-carboxamide

N-(2-methyl-5-(3-(trifluoromethyl)phenylcarbamoyl)phenyl)-4-

(memylsulfinyl)mieno[3,2-d]pyrimidine-7-carboxamide (30mg, 0.060mmol) was dissolved in dioxane (ImL) and DIPEA (20μί, 0.119mmol) and 4-methoxybenzeneamine (37mg, 0.298mmol) were added thereto. The reaction mixture was stirred at 120°C for 18 hours, cooled to room temperature, dilluted with ethyl acetate and washed with a brine. The organic layer was dried with MgS0 ₄ , filtered and concentrated under reduced pressure. The resulting mixture was purified by silica gel chromatography to obtain the title compound (28mg, 81% yield).

Ή NMR (400 MHz, DMSO-de) δ 11.85 (s, 1H), 10.56 (s, 1H), 10.06 (s, 1H), 9.00 (s, 1H), 8.87 (s, 1H), 8.73 (s, 1H), 8.26 (s, 1H), 8.08 (d, 1H), 7.72 (d, 1H), 7.63-7.57 (m, 3H), 7.47 (d, 2H), 7.01 (d, 2H), 3.79 (s, 3H), 2.54 (s, 3H); MS m/z [M+l] 578.07.

Example 14: N-(2-methyI-5-(3-(trifluoromethyl)phenylcarbamoyl)phenyl)-4- (3,4,5- trimethoxyphenylamino)thieno [3,2-d] pyrimidine-7-carboxamide The procedure of Step 3 of Example 13 was repeated except for using the compound obtained in Step 2 of Example 13 and 3,4,5-trimethoxyaniline to obtain the title compound (see Table 1).

Example 15 : N-(2-methyl-5-(3-(trifluoromethyl)phenylcarbamoyl)phenyl)-4- (6- methylpyridine-3-y lamino)thieno [3,2-d] pyrimidine-7-carboxamide

The procedure of Step 3 of Example 13 was repeated except for using the compound obtained in Step 2 of Example 13 and 6-methylpyridine-3-amine to obtain the title compound (see Table 1). Example 16: 4-(4-(4-Ethylpiperazin-l-yl)phenyIamino)-N-(2-methyI-5-(3-

(trifluoromethyl)phenylcarbamoyl)phenyl)thieno[3,2-d]pyri midine-7-carboxamide

The procedure of Step 3 of Example 13 was repeated except for using the compound obtained in Step 2 of Example 13 and 4-(4-ethylpiperazine-l-yl)aniline to obtain the title compound (see Table 1).

Example 17: 4-(Isopropylamino)-N-(2-methyl-5-(3-

(trifluoromethyl)phenylcarbamoyl)phenyl)thienof3,2-d]pyri midine-7-carboxamide

The procedure of Step 3 of Example 13 was repeated except for using the compound obtained in Step 2 of Example 13 and isopropylamine to obtain the title compound (see Table 1). Example 18: N-(2-methyl-5-(3-(trifluoromethyI)phenylcarbamoyl)phenyl)-4- (methy lamino)thieno [3,2-d] pyrimidine-7-car boxamide

The procedure of Step 3 of Example 13 was repeated except for using the compound obtained in Step 2 of Example 13 and methylamine hydrochloride to obtain the title compound (see Table 1).

Example 19: 4-(2-Hydroxyethylamino)-N-(2-methyl-5-(3-

(trifluoromethyl)phenylcarbamoyl)phenyl)thieno[3,2-d]pyri midine-7-carboxamide The procedure of Step 3 of Example 13 was repeated except for using the compound obtained in Step 2 of Example 13 and 2-aminoethanol to obtain the title compound (see Table 1).

Example 20: N-(2-Methyl-5-(3-(trifluoromethyl)phenylcarbamoyl)phenyl)-4- (2- morpholinoethylamino)thieno [3,2-d] pyrimidine-7-carboxamide

The procedure of Step 3 of Example 13 was repeated except for using the compound obtained in Step 2 of Example 13 and 2-morpholino ethaneamine to obtain the title compound (see Table 1). The compound of Preparation Example 3 was prepared according to Reaction Scheme 7 below:

Reaction Scheme 7

Preparation Example 3: 4-Chlorothieno[3,2-d]pyrimidine-7-carboxylic acid

Step 1 : 7-Methylthieno[3,2-d]pyrimidine-4(3H)-one

Methyl 3-amino-4-methylthiophene-2-carboxylate (3.0g) was added to formamide (5mL), and the mixture was stirred at 200°C for 3 hours. After cooling to room temperature water (50mL) was added thereto and then stirred for 15 hours at room temperature to obtain a solid. The solid was filtered, washed with water and dried with N ₂ gas to obtain the title compound as a white solid without further purification.

1H NMR (400MHz, DMSO-d ₆ ) δ 12.50 (br, 1H), 8.16 (s, 1H), 7.86 (s, 1H), 2.30 (s,

3H).

Step 2: 4-Chloro-7-methylthieno[3,2-d]pyrimidine

7-Methylthieno[3,2-d]pyrimidine-4(3H)-one (1.5g) was added to POCl ₃ (lOmL), and the mixture was stirred at 110°C for 2 hours. The reaction mixture was cooled to room temperature, concentrated and dilluted with DCM, and sat. NaHC0 ₃ solution was added thereto. The aqueous layer was extracted with DCM to combine organic layers. The organic layer was dried with MgS0 ₄ and filtered to obtain the title compound as a white solid.

1H NMR (400MHz, DMSO-d ₆ ) δ 9.06 (s, 1H), 8.24 (s, 1H), 2.43 (s, 3H). Step 3 : -(Bromomethyl)-4-chlorothieno[3 ,2-d]pyrimidine

4-Chloro-7-methylthieno[3,2-d]pyrimidine (6.62g, 33.0mmol) and NBS (5.87g, 33.0mmol) were dissolved in CCL^lOOmL) and benzoylperoxide (l.Og, 80% purity) was added thereto. The reaction mixture was stirred at 100°C for an hour, cooled to room temperature, filtered with Celite and concentrated to obtain the title compound having a yellow color without further purification.

Ή NMR (400MHz, DMSO-d ₆ ) δ 9.15 (s, 1H), 8.75 (s, 1H), 4.93 (s, 2H).

Step 4: -Chlorothieno[3,2-d]pyrimidine-7-yl)methyl acetate

7-(Bromomethyl)-4-chlorothieno[3,2-d]pyrimidine (9.17g, 33.0mmol) was dissolved in DMF (54mL) and then sodium acetate (27g, 330mmol) and potassium iodide (10.96g, 66.0mmol) were added thereto. The reaction mixture was stirred at 35°C for 4 hours, dilluted with ethyl acetate and washed with sat. sodium thiosulfate solution 5 times. The reaction mixture was dried with MgS0 ₄ , filtered, concentrated and purified by silica gel chromatography to obtain to obtain the title compound as a white solid.

1H NMR (400MHz, DMSO-d ₆ ) δ 9.06(s, 1H), 8.60(s, 1H), 5.38(s, 2H), 2.06(s, 3H).

Step 5: 4-Chlorothieno[3,2-d]pyrimidine-7-yl)methanol

(4-Chlorothieno[3,2-d]pyrimidine-7-yl)methyl acetate (3.12g, 12.06mmol) was dissolved in THF (240mL) and then IN NaOH solution (lOOmL) was added thereto at room temperature. The reaction mixture was stirred for 30 minutes, dilluted with ethyl acetate, and the aqueous layer was extracted with ethyl acetate. The organic layer was dried with MgS0 ₄ , filtered and concentrated to obtain the title compound as a white solid without further purification.

1H NMR (400MHz, DMSO-d ₆ ) δ 9.09(s, 1H), 8.33(s, 1H), 5.47(t, 1H), 4.84(d, 2H).

Step 6: -Chlorothieno[3,2-d]pyrimidine-7-carbaldehyde

(4-Chlorothieno[3,2-d]pyrimidine-7-yl)methanol (215mg, 0.99mmol) and Mn0 ₂ (863mg, 9.9mmol, lOeq) were dissolved in chloroform (lOmL) and stirred at 70°C for 2 hours. The reaction mixture was cooled to room temperature, filtered with Celite and concentrated to obtain the title compound as a white solid without further purification.

1H NMR (400MHz, DMSO-d ₆ ) δ 10.30(s, 1H), 9.40(s, 1H), 9.20(s, 1H).

Step 7: 4-CMorothieno[3,2-d]pyrimidine-7-carboxylic acid

NaH ₂ P0 ₄ .2H ₂ 0 (170mg, 1.07mmol) was dissolved in water (0.6mL) and 4- chlorothieno[3,2-d]pyrimidine-7-carbaldehyde (lOOmg, 0.47mmol) dissolved in DMSO (4.8mL) was added thereto at 10°C. The reaction mixture was cooled to 0°C and NaC10 ₂ (170mg, 1.88mmol) dissolved in water (0.6mL) was added thereto at 0°C. The reaction mixture was heated to room temperature, stirred for 2 hours, and water (lOmL) was added thereto. The reaction mixture was stirred for 2 hours and then filtered. The filtrate was concentrated to obtain the title compound as a white solid.

1H NMR (400MHz, DMSO-d ₆ ) δ 9.07(s, 1H), 8.87(s, 1H).

The compound of Example 21 was prepared according to Reaction Scheme 8 below: Reaction Scheme 8

Example 21: 4-(Cyclopropylamino)-N-(2-methyl-5-(3- (trifluoromethyl)phenylcarbamoyl)phenyl)thieno [3,2-d] pyrimidine-7-carboxamide

Step 1: Ethyl 3-(4-chlorothieno[3,2-d]pyrimidine-7-carboxamido)-4-methyl benzoate

The procedure of Step 1 of Example 8 was repeated except for using the compound obtained in Preparation Example 3 as a starting material to obtain the title compound.

MS m/z [M+l] 376.05.

Step 2: Ethyl 3-(4-(cyclopropylamino)thieno[3,2-d]pyrimidine-7-carboxamido )-4-methyl benzoat

The procedure of Step 3 of Example 13 was repeated except for using the compound obtained in Step 1 of Example 21 as a starting material to obtain the title compound.

Ή NMR (400 MHz, DMSOd ₆ ) δ 11.91 (s, 1H), 8.97 (s, 1H), 8.66 (s, 1H), 8.51 (s, 1H), 7.66 (d, 1H), 7.44 (d, 1H), 4.32 (q, 2H), 3.08-3.03 (m, 1H), 2.54 (s, 3H), 1.33 (t, 3H), 0.86 (br s, 2H), 0.70 (br s 2H) ; MS m/z [M+l] 397.93.

Step 3 : 3-(4-(Cyclopropylamino)thieno[3,2-d]pyrimidine-7-carboxamido )-4- methylbenzoic acid

The procedure of Step 4 of Example 1 was repeated except for using the compound obtained in Step 2 of Example 21 as a starting material to obtain the title compound.

1H NMR (400 MHz, DMSO-d ₆ ) δ 13.02-12.80 (m, 1H), 11.90 (s, 1H), 8.96 (s, 1H), 8.93 (s, 1H), 8.66 (s, 1H), 8.53 (s, 1H), 7.64 (dd, 1H), 7.40 (d, 1H), 3.07-3.03 (m, 1H), 2.54 (s, 3H), 0.86 (br s, 2H), 0.70 (br s, 2H). Step 4: 4-(Cyclopropylamino)-N-(2-methyl-5-(3-

(trifluoromemyl)phenylcarbamoyl)phenyl)mieno[3,2-d]pyrimi dine-7-carboxamide

The procedure of Step 5 of Example 1 was repeated except for using the compound obtained in Step 3 of Example 21 as a starting material to obtain the title compound.

MS m/z [M+l] 512.06

The compound of Preparation Example 4 was prepared according to Reaction Scheme 9 below: Reaction Scheme 9 n02

Preparation Example 4: 4-(Cyclopropylamino)thieno[3,2-d)pyrimidine-7-carboxylic acid

Step 1: (4-(Cyclopropylamino)thieno[3,2-d]pyridine-7-yl)methanol

Isopropanol (590mL) and cyclopropylamine (102mL, 1.5mol) were added to the compound (59g, 29.3mmol) obtained in Step 5 of Preparation Example 3, and stirred for 12 hours with reflux. The reaction mixture was cooled to room temperature, concentrated, dilluted with ethyl acetate (1L), washed with a sat. aqueous sodium hydrogen carbonate solution (800mL x 3) and then concentrated. The resulting mixture was crystallized using a methanol/diethyl ether/hexane mixture (1:10:10, v/v/v) to obtain the title compound (46.5g).

1H-NMR (300MHz, DMSO-d ₆ ) δ 8.42 (s, 1H), 7.93 (d, 1H), 7.84 (s, 1H), 5.23-5.19 (t, 1H), 4.70 (d, 2H), 2.96 (m, 1H), 0.78 (m, 2H), 0.62 (m, 2H).

Step 2: 4-(Cyclopropylamino)thieno[3,2-d]pyrimidine-7-carbaldehyde

The compound (46.5g, 210mmol) obtained in Step 1 of Preparation Example 4 was dissolved in chloroform (2.0L), manganese dioxide (183g, 2.1mol) was added thereto and stirred for 12 hours with reflux. The reaction mixture was cooled to room temperature, manganese dioxide was filtered with Celite, and the filtrate was concentrated and dried to obtain the title compound (40.9g) without further purification.

Ή-NMR (300MHz, DMSO-d ₆ ) δ 10.3 (s, 1H), 8.96 (s, TH), 8.55 (s, 1H), 8.30 (d, 1H), 2.99 (m, 1H), 0.80 (m, 2H), 0.65 (m, 2H).

Step 3: 4-(Cyclopropylamino)thieno[3,2-d]pyrimidine-7-carboxylic acid

NaH ₂ P0 ₄ .2H ₂ 0 (67g, 429mmol) was dissolved in water (220mL), the compound (41g, 187mmol) obtained in Step 2 of Preparation Example 4 dissolved in a acetone/dimethyl sulfoxide mixture (1 :1, v/v) (640mL) was added thereto slowly, cooled to 0°C and stirred for 30 minutes. NaC10 ₂ (25.3g, 280mmol) dissolved in water (220mL) was added to the reaction mixture slowly at 0°C and stirred at room temperature for 5 hours. Water (400mL) was added to the reaction mixture to remove gel therein, and the resulting solid was filtered and washed sequentially with water, ethyl acetate and dimethyl ether. Ethyl acetate (400mL) was added thereto and stirred to remove the remaining dimethyl sulfoxide solvent. The solid was filtered and washed with diethyl ether to obtain the title compound (32g).

1H-NMR (300MHz, DMSO-d ₆ ) δ 8.97 (s, 1H), 8.75 (br. s, 1H), 8.59 (s, 1H), 3.05 (m, 1H), 0.86 (m, 2H), 0.69 (m, 2H).

The compound of Preparation Example 5 was prepared according to Reaction Scheme 10 below: Reaction Scheme 10

Preparation Example 5: 4-(l-Methylpiperidine-4-yloxy)-3- (trifluoromethyl)benzeneamine

Step 1: Methyl 4-hydroxy-3-(trifluoromethyl)benzoate

4-Hydroxy-3-(trifluoromethyl) benzoic acid (2g, 9.7mmol) was dissolved in methanol (40mL) and 12N HC1 (10 drops) was added thereto. The reaction mixture was stirred at 90°C for 48 hours, cooled to room temperature and concentrated, and then ethyl acetate (lOOmL) was added thereto. The reaction mixture was washed with sat. NaHC0 ₃ solution, and the organic layer was dried with Na ₂ S0 ₄ , filtered and concentrated to obtain the title compound (2.07g, 91%) without further purification.

1H NMR (300MHz, DMSO-d ₆ ) δ 11.60 (s, 1H), 8.02 (m, 2H), 7.10 (d, 1H), 4.26 (q,

2H), 1.28(t, 3H).

Step 2: Methyl 4-(l-methylpiperidine-4-yloxy)-3-(trifluoromethyl)benzoate

In a round bottom flask wrapped with foil, methyl 4-hydroxy-3- (trifluoromethyl)benzoate (500mg, 2.13mmol) was dissolved in methylene chloride (lOmL). 4-Hydroxy-l-methyl-piperidine (0.25mL, 2.13mmol), PPh ₃ (670mg, 2.56mmol), and DIAD (0.50mL, 2.56mmol) were added thereto, and the mixture was stirred at room temperature for 23 hours. DTAD (4.26mmol) and PPh ₃ (1.12g, 4.26mmol) were added at 0°C to the reaction mixture, which was stirred at room temperature for 24 hours. The resulting mixture was concentrated and then purified by silica gel chromatography (Hex/EA=l/l→DCM/MeOH=10/l) to obtain the title compound (512mg, 76%).

1H NMR (300MHz, DMSC ¾ δ 8.14 (dd, 1H), 8.08 (d, 1H), 7.43 (d, 1H), 4.77 (m,

1H), 4.30(q, 2H), 2.35 (m, 2H), 2.24(m, 2H), 2.19 (s, 3H), 1.93 (m, 2H), 1.73 (m, 2H), 1.27 (t, 3H).

Step 3: 4-(l-Methylpiperidine-4-yloxy)-3-(trifluoromethyl)benzoic acid

Methyl 4-(l -methylpiperidine-4-yloxy)-3-(trifluoromethyl)benzoate (512mg, 1.61mmol) was dissolved in a mixture of THF (1.5mL), MeOH (1.5mL) and H ₂ 0 (1.5mL), and then NaOH (193mg, 4.84mmol) was added thereto. The reaction mixture was stirred at 90°C for 22 hours and cooled to room temperature, and IN HCl was added dropwise thereto to pH 4. The reaction mixture was concentrated, and a mixed solvent (DCM/MeOH=5/l) was added thereto. The remaining solid was filtered, and the filtrate was concentrated to obtain the title compound (391mg, 80%) without further purification.

1H NMR (300MHz, DMSO-<¾ 5 8.16 (dd, 1H), 8.11 (d, 1H), 7.46 (d, 1H), 4.93 (m, 1H), 3.03(m, 4H), 2.63 (s, 3H), 2.15(m, 2H), 1.95 (m, 2H). Step 4: t-Butyl 4-(l-methylpiperidine-4-yloxy)-3-(trifluoromethyl)phenylcarb amate

4-(l -Methylpiperidine-4-yloxy)-3-(trifluoromethyl)benzoic acid (200mg, 0.659mmol) was dissolved in t-BuOH (6mL) and then DPPA (0.2 ImL, 0.989mmol) and TEA (0.14mL, 0.989mmol) was added thereto. The reaction mixture was stirred at 90°C for 23 hours, cooled to room temperature, and sat. NaHC0 ₃ solution was added thereto. The aqueous layer was washed with ethyl acetate, and the combined organic layer was dried with Na ₂ S04, filtered and concentrated. The resulting mixture was purified by silica gel chromatography (DCM/MeOH=10/l) to obtain the title compound (82mg, 45%).

1H NMR (300MHz, DMSO-</ ₆ ) δ 9.44 (s, 1H), 7.79 (s, 1H), 7.56 (d, 1H), 7.20 (d, 1H), 4.55(m, 1H), 2.66 (m, 2H), 2.38 (m, 2H), 2.31 (s, 3H), 1.93 (m, 2H), 1.74 (m, 2H), 1.46 (s, 9H).

Step 5: -(l-Methylpiperidine-4-yloxy)-3-(trifluoromethyl)benzeneamin e

t-Butyl 4-(l -methylpiperidine-4-yloxy)-3-(trifluoromethyl)phenylcarbamat e (82mg, 0.299mmol) was dissolved in methylene chloride (ImL), and TFA (ImL) was added thereto. The reaction mixture was stirred at room temperature for 2 hours and then concentrated. The reaction mixture was dilluted with ethylene chloride and washed with sat. NaHC0 ₃ solution. The organic layer was dried with Na ₂ S0 ₄ , filtered and concentrated to obtain the title compound (63mg, 77%) without further purification.

1H NMR (300MHz, OMSO-d ₆ ) δ 6.97 (dd, 1H), 6.79 (d, 1H), 6.73 (d, 1H), 5.00 (s, 2H), 4.26(m, 1H), 2.50 (m, 2H), 2.13 (s, 3H), 2.10 (m, 2H), 1.86 (m, 2H), 1.61 (m, 2H).

The compound of Preparation Example 6 was prepared according to Reaction Scheme 11 below: Reaction Scheme 11

Preparation Example 6: 3-Amino-4-methyl-N-(3-(2-methyl-lH-imidazole-l-yl)-5- (trifluoromethyl)phenyl)benzamide

Step 1: -(2-Methyl-lH-imidazole-l-yl)-5-(trifluoromethyl)aniline

3-Bromo-5-(trifluoromethyl)aniline (1.2mL, 8.60mmol) and 2-methylimidazole

(847mg, 10.3mmol) were dissolved in dimethyl sulfoxide (8.6mL), and Cul (490mg, 2.58mmol), 8-quinolinol (370mg, 2.58mmol) and potassium carbonate (2.38g, 17.2mmol) were added thereto. The reaction mixture was treated with N ₂ gas for 10 minutes to remove gas and subjected to a reaction at 120°C for 12 hours. The reaction mixture was cooled to room temperature, and 14% aqueous ammonium hydroxide solution was added thereto. The reaction mixture was stirred for 1 hours at room temperature, dilluted with ethyl acetate (30mL) and washed with an aqueous sodium hydrogen carbonate solution (30mL). The organic layer was washed with water, dried with sodium sulfate, and the solvent was distilled under reduced pressure. The concentrated mixture was filtered with a silica column using ethyl acetate to remove copper impurities, and the filtrate was distilled under reduced pressure. The concentrated mixture was crystallized with ethyl acetate (3mL) and n-hexane (30mL) to obtain the title compound (1.31g) as a bright green solid..

1H NMR (300 MHz, DMSO-d ₆ ) δ 7.27 (s, 1H), 6.90 (s, 2H), 6.78 (s, 2H), 5.93 (s, 2H), 2.26 (s, 3H).

Step 2: 4-Methyl -N-(3-(2-methyl-lH-imidazole-l-yl)-5-(trifluoromethyl)phenyl )-3- nitrobenzamide

3-Nitro-4-methyl benzoic acid (14.8g, 81.5mmol) was dissolved in DMF (150mL), and HATU (62g, 163mmol) and DIPEA (57mL, 326mmol) were added thereto at room temperature. The reaction mixture was stirred for 15 minutes, 3-(2-methyl-lH-imidazole- l-yl)-5-(trifluoromethyl)aniline (13. lg, 54.3mmol) was added thereto and stirred at 45°C for 16 hours. The reaction mixture was cooled to room temperature, diluted with ethyl acetate and washed with sat. NaHC0 ₃ solution. The organic layer was dried with anhydrous MgS0 ₄ , filtered with Celite and concentrated. The residue was crystallized with ethyl acetate and hexane to obtain the title compound (27g).

Ή NMR (300 MHz, DMSO-d ₆ ) δ 10.9 (s, 1H), 8.61 (s, 1H), 8.25-8.22 (m, 2H), 8.16 (s, 1H), 7.74-7.71 (d, 1H), 7.62 (s, 1H), 7.41 (s, 1H), 6.95 (s, 1H), 2.55 (s, 3H), 2.34 (s, 3H). Step 3: 3-Amino-4-methyl-N-(3-(2-methyl-lH-imidazole-l-yl)-5-

(trifluoromethyl)phenyl)benzamide

The compound (27g, 72.1mmol) obtained in Step 2 of Preparation Example 6 was dissolved in ethanol (300mL), SnCl ₂ -2H ₂ 0 (81g, 360mmol) was added thereto and stirred at 80°C for 3 hours. The reaction mixture was cooled to room temperature and diluted with ethyl acetate. The organic layer was washed with sat. NaHC0 ₃ solution several times. The organic layer was dried with anhydrous MgS0 ₄ , filtered and concentrated to obtain the title compound without further purification.

MS m/z [M+l] 375.25.

Example 22: 4-(Cyclopropylammo)-N-(2-methyI-5-(3-(2-methyl-lH-imida2oIe- l-yl)-5- (trifluoromethyl)phenylcarbamoyl)phenyl)thieno[3,2-d]pyrimid me-7-carboxamide

The compound (7.2g, 31mmol) obtained in Step 3 of Preparation Example 4 was dissolved in DMF (lOOmL) and then HATU (25g, 65.4mmol) and DIPEA (23mL, 131mmol) were added thereto at room temperature. The reaction mixture was stirred for 15 minutes, 3-amino-4-methyl-N-(3-(2-methyl-lH-imidazole-l-yl)-5-

(trifluoromethyl)phenyl)benzamide (8.2g, 22mmol) obtained in Step 3 of Preparation Example 6 was added thereto and stirred at 45°C for 16 hours. The reaction mixture was cooled to room temperature, diluted with ethyl acetate and washed with sat. NaHC0 ₃ solution. The organic layer was dried with anhydrous MgS0 ₄ , filtered with Celite and concentrated. The residue was crystallized with ethyl acetate and hexane to obtain the title compound (5g).

Ή NMR (DMSO-d ₆ ) δ 11.9 (s, IH), 10.8 (s, IH), 8.98 (s, IH), 8.90 (s, IH), 8.67 (s, IH), 8.52 (s, IH), 8.31 (s, IH), 8.18 (s, IH), 7.73 (d, IH), 7.58 (s, IH), 7.50 (d, IH), 7.42 (s, IH), 6.95 (s, IH), 3.04 (m, IH), 2.55 (s, 3H), 2.35 (s, 3H), 0.84 (m, 2H), 0.69 (m, 2H).

The compound of Preparation Example 7 was prepared according to Reaction Scheme 12 below:

Reaction Scheme 12

Preparation Example 7: 4-(3-Amino-4-methylbenzamido)-2-(trifluoromethyl)benzyl acetate Step 1 : -(Bromomethyl)-4-nitro-2-(trifluoromethyl)benzene

The procedure of Step 3 of Preparation Example 3 was repeated except for using 1 - methyl-4-nitro-2-(trifluoromethyl)benzene as a starting material to obtain the title compound.

MS m/z [M+l] 283.09, 285.97.

Step 2: 4-Nitro-2-(trifluoromethyl)benzyl acetate

The procedure of Step 4 of Preparation Example 3 was repeated except for using the compound obtained in Step 1 of Preparation Example 7 as a starting material to obtain the title compound.

MS m/z [M+l] 264.05. Step 3: 4-Amino-2-(trifluoromethyl)benzyl acetate

The procedure of Step 2 of Example 1 was repeated except for using the compound obtained in Step 2 of Preparation Example 7 as a starting material to obtain the title compound.

Ή NMR (400 MHz, DMSO-d ₆ ) δ 7.24 (d, 1H), 6.91 (d, 1H), 6.76 (dd, 1H), 5.71 (s, 2H), 4.99 (s, 2H), 2.00 (s, 3H)

Step 4: -(4-Methyl-3-nitrobenzamido)-2-(trifluoromethyl)benzyl acetate

The procedure of Step 2 of Preparation Example 6 was repeated except for using the compound obtained in Step 3 of Preparation Example 7 as a starting material to obtain the title compound.

MS m/z [M+l] 397.10.

Step 5: -(3-Amino-4-methylbenzamido)-2-(trifluoromethyl)benzyl acetate

The procedure of Step 3 of Preparation Example 6 was repeated except for using the compound obtained in Step 4 of Preparation Example 7 as a starting material to obtain the title compound.

Ή NMR (400 MHz, DMSO-d ₆ ) δ 10.36 (s, IH), 8.27 (s, IH), 8.08 (d, IH), 7.62 (d, IH), 7.18 (s, IH), 7.08 (q, 2H), 5.17 (s, 2H), 5.11 (s, 2H), 2.12 (s, 3H), 2.07 (s, 3H); MS m/z [M+l] 368.98.

The compounds of Examples 23 to 26 were prepared according to Reaction Scheme 13 below:

Reaction Scheme 13

Example 23 . 4-(3-(4-(Cyclopropylammo)thieno[3,2-d]pyrimidine-7-carboxami do)- 4-methylbenzamido)-2-(trifluoromethyl)benzyl acetate

The procedure of Example 22 was repeated except for using the compound obtained in Preparation Example 7 instead of 3-amino-4-methyl-N-(3-(2-methyl-lH- imidazole-l-yl)-5- (trifluoromethyl)phenyl)benzamide to obtain the title compound.

1H NMR (400 MHz, DMSO-d ₆ ) δ 11.91 (s, IH), 10.60 (s, IH), 8.99 (br s, IH), 8.89 (s, IH), 8.69 (br s, IH), 8.53 (br s, IH), 8.29 (s, IH), 8.12 (d, IH), 7.71 (d, IH), 7.65 (d, IH), 7.18 (d, IH), 5.19 (s, 2H), 3.06-3.04 (m, IH), 2.89 (s, 3H), 2.08 (s, 3H), 0.85-0.80 (m, 2H), 0.70-0.64 (m, 2H); MS m/z [M+l] 584.04.

Example 24: 4-(Cyclopropylainmo)-N-(5-(4-(hydroxymetliyl)-3-

(trifluoromethyl)phenylcarbamoyl)-2-methylphenyl)thieno[3 ,2-d]pyrimidine-7- carboxamide

The procedure of Step 5 of Preparation Example 3 was repeated except for using the compound obtained in Example 23 instead of (4-chlorothieno[3,2-d]pyrimidine-7- yl)methyl acetate to obtain the title compound.

1H NMR (400 MHz, DMSO-d ₆ ) δ 11.91 (s, IH), 10.50 (s, IH), 8.99 (br s, IH), 8.88

(s, IH), 8.68 (br s, IH), 8.52 (br s, IH), 8.20 (s, IH), 8.08 (d, IH), 7.72 (dd, 2H), 7.47 (d, IH), 5.44 (br s, IH), 4.64 (d, 2H), 3.09-3.05 (m, IH), 2.70 (s, 3H), 0.92-0.80 (m, 2H), 0.76-0.65 (m, 2H) ; MS m/z [M+l] 541.96. Example 25: 4-(3-(4-(Cyclopropylammo)thieno[3,2-d]pyriinidine-7-carboxam ido)-4- methylbenzamido)-2-(trifluoromethyl)benzyI methanesulfonate

4-(Cyclopropylamino)-N-(5-(4-(hydroxymethyl)-3- (trifluoromethyl)phenylcarbamoyl)-2-methylphenyl)thieno[3,2- d]pyrimidine-7- carboxamide (300mg, 0.554mmol) was dissolved in DCM (2.5mL), and TEA (0.15mL, 1.1 lmmol) was added thereto. Methanesulfonyl chloride (66μΙ_<, 0.83mmol) was added to the reaction mixture at 0°C. The reaction mixture was stirred for 6 hours and diluted with DCM (lOmL). The organic layer was washed with brine, dried with MgS0 , filtered and concentrated under reduced pressure to obtain the title compound without further purification.

1H NMR (400 MHz, DMSO-d ₆ ) δ 11.92 (s, 1H), 10.63 (s, 1H), 8.99 (br s, 1H), 8.89 (s, 1H), 8.68 (br s, 1H), 8.52 (br s, 1H), 8.28 (s, 1H), 8.14 (d, 1H), 7.72 (dd, 2H), 7.49 (d, 1H), 4.86 (s, 2H), 3.12-3.05 (m, 1H), 2.67 (s, 3H), 1.20 (s, 3H), 0.92-0.87 (m, 2H), 0.74- 0.60 (m, 2H); MS m/z [M+l] 619.98

Example 26 : 4-(Cyclopropylamino)-N-(2-methyl-5-(4-((4-metliyl- 1 H-imidazole- 1- yl)methyl)-3-(trifluoromethyl)phenylcarbamoyl)phenyl)thieno[ 3,2-d]pyrimidine-7- carboxamide

4-(3-(4-(Cyclopropylamino)thieno[3,2-d]pyrimidine-7-carboxam ido)-4- methylbenzamido)-2-(trifluoromethyl)benzyl methanesulfonate (30mg, 0.048mmol) was dissolved in DMF (lmL), K ₂ CO ₃ (20mg, 0.145mmol) and 4-methyl-l H-imidazole (20mg, 0.242mmol) were added thereto and stirred at room temperature for 18 hours. The reaction mixture was diluted with ethyl acetate (lOmL) and washed with brine. The organic layer was dried with MgS0 ₄ , filtered and concentrated under reduced pressure. The resulting mixture was purified by silica gel chromatography (MeOH/DCM=5/95) to obtain the title compound (23mg, 78% yield).

Ή NMR (400 MHz, DMSO-d ₆ ) δ 11.94 (s, 1H), 10.74 (s, 1H), 8.99 (br s, 1H), 8.93 (s, 1H), 8.68 (br s, 1H), 8.53 (br s, 1H), 8.41 (s, 1H), 8.21 (s, 1H), 7.82 (s, 1H), 7.74 (d, 2H), 7.51 (d, 1H), 4.47 (s, 2H), 3.09-3.02 (m, 1H), 2.24 (s, 3H), 1.17-1.00 (m, 2H), 0.85-0.78 (m, 2H).

Example 27: 4-(CyclopropyIaimno)-N-(2-methyl-5-(3-(4-methyl-lH-imidazoIe -l-yI)-5- (trifluoromethyl)phenylcarbamoyl)phenyl)thieno[3,2-d]pyrimid ine-7-carboxamide

The procedure of Step 5 of Example 1 was repeated except for using 3-(4- (cyclopropylamino)thieno[3,2-d]pyrimidine-7-carboxamido)-4-m ethylbenzoic acid obtained in Step 3 of Example 21 and 3-(4-methyl-lH-imidazole-l-yl)-5- (trifluoromethyl)aniline to obtain the title compound (see Table 1).

Example 28 : N-(5-(3-bromo-5-(trifluoromethylcarbamoyl)plienyl)-2-methylp lienyl)-4- (cyclopropylamino)thieno [3,2-d] pyrimidine-7-carboxamide The procedure of Step 5 of Example 1 was repeated except for using 3-(4-

(cyclopropylamino)tlueno[3,2-d]pyrimidme-7-carboxamido)-4 -methylbenzoic acid obtained in Step 3 of Example 21 and 3-bromo-5-(trifluoromethyl)aniline to obtain the title compound (see Table 1). Example 29: 4-(Cyclopropylamino)-N-(2-methyl-5-(6-morpholinopyridine-3- ylcarbamoyl)phenyl)thieno[3,2-d]pyrimidine-7-carboxamide

The procedure of Step 5 of Example 1 was repeated except for using 3-(4- (cyclopropylamino)thieno[3,2-d]pyrimidine-7-carboxamido)-4-m ethylbenzoic acid obtained in Step 3 of Example 21 and 6-morpholinopyridine-3 -amine to obtain the title compound (see Table 1).

Example 30 : 4-(Cyclopropylainino)-N-(5-(6-(4-ethylpiperazine- l-yl)pyridine-3- ylcarbamoyl)-2-methylphenyl)thieno [3,2-d] pyrimidine-7-carboxamide The procedure of Step 5 of Example 1 was repeated except for using 3-(4- (cyclopropylamino)thieno[3,2-d]pyrimidine-7-carboxamido)-4-m ethylbenzoic acid obtained in Step 3 of Example 21 and 6-(4-ethylpiperazine-l-yl)pyridine-3 -amine to obtain the title compound (see Table 1).

Examples 31, and 34 to 38

The procedure of Example 22 was repeated except for using an appropriate starting compound (see Table 1).

Examples 32 and 33

The procedure of Example 26 was repeated except for using an appropriate starting compound (see Table 1).

The compound of Example 39 was prepared according to Reaction Scheme 14 below:

Reaction Scheme 14

Example 39: 4-Amino-N-(2-methyl-

(trifluoromethyl)benzamido)phenyl)thieno[3,2-d]pyrimidine -7-carboxamide

Step 1: 4-Aminothieno[3,2-d]pyrimidine-7-carboxylic acid

The procedures of Steps 2 and 3 of Preparation Example 2 was repeated except for using the compound obtained in Step 2 of Example 1 as a starting material to obtain the title compound.

MS m/z [M+l] 196.03.

Step 2: 4-Amino-N-(2-memyl-5-nitrophenyl)thieno[3,2-d]pyrimidine-7-c arboxamide

The procedure of Example 8 was repeated except for using the compound obtained in Step 1 of Example 39 as a starting material to obtain the title compound.

Ή NMR (300 MHz, DMSO-d ₆ ) δ 12.12 (s, 1H), 9.26 (d, 1H), 8.95 (s, 1H), 8.61 (s, 1H), 7.85 (dd, 1H), 7.84 (or s, 2H), 7.55 (d, 1H), 2.60 (s, 3H). MS m/z [M+l] 330.06.

Step 3 : 4-Amino-N-(5-amino-2-methylphenyl)thieno[3,2-d]pyrimidine-7- carboxamide

The compound (lg, 3.0mmol) obtained in Step 2 of Example 39 was added to methanol (15mL) and Pd/C (200mg), and stirred under H ₂ atmosphere for 14 hours at room temperature. The reaction mixture was filtered using Celite and concentrated to obtain the title compound (152mg).

1H NMR (300 MHz, DMSO-d ₆ ) δ 11.48 (s, 1H), 8.86 (s, 1H), 8.54 (s, 1H), 7.88 (br s, 2H), 7.59 (d, 1H), 6.88 (d, 1H), 6.27 (dd, 1H), 4.92 (br s, 2H), 2.27 (s, 3H). MS m/z [M+l] 300.09.

Step 4: 4-Amino-N-(2-methyl-5-(3-(trifluoromethyl)benzamido)phenyl)t hieno[3,2- d]pyrimidine-7-carboxamide

4-Amino-N-(5-arrdno-2-methylphenyl)thieno[3,2-d]pyrimidine-7 -carboxam

(30mg, O.lmmol) was dissolved in THF (ImL) and ΤΕΑ(42μί, 0.30mmol) was added thereto. 3-(trifluoromethyl)benzoyl chloride (19μΕ, 0.13mmol) was added to reaction mixture at 0°C and stirred at room temperature for 6 hours. The reaction mixture was diluted with ethyl acetate (lOmL) and washed with brine. The organic layer was dried with MgS0 ₄ , filtered and concentrated under reduced pressure. The resulting mixture was purified by silica gel chromatography (3/97 MeOH/DCM) to obtain the title compound (35mg, 74% yield).

1H NMR (300 MHz, DMSO-d ₆ ) δ 11.74 (s, 1 H), 10.51 (s, 1 H), 8.92 (s, 1 H), 8.67

(s, 1 H), 8.58 (s, 1 H), 8.31 (s, 1 H), 8.28 (d, 1 H), 7.96 (d, 1 H), 7.94 (s, 2 H), 7.77 (t, 1 H), 7.58 (d, 1 H), 7.27 (d, 1 H), 2.44 (s, 3 H).

The compound of Preparation Example 8 was prepared according to Reaction Scheme 15 below:

Reaction Scheme 15

Preparation Example 8: N-(3-amino4-methylphenyl)-4-morpholino (trifluoromethyl)benzamide

Step 1 : 4-Morpholino-3-(trifluoromethyl)benzonitrile

4-Fluoro-3-(trifluoromethyl)benzonitrile (2.0g, 10.58mmol) was dissolved in dimethyl acetamide (35mL), morpholine (2.88mL, 31.73mmol) was added thereto, and the mixture was subjected to a reaction at 145°C for 19 hours. The reaction mixture was cooled to room temperature, diluted with ethyl acetate (50mL), and the organic layer was washed with an aqueous sodium hydrogen carbonate solution (50mL). The organic layer was washed with water, dried with sodium sulfate, and the solvent was distilled under reduced pressure. The concentrated mixture was crystallized with ethyl acetate (lOmL) and n-hexane (80mL) to obtain the title compound as a brown solid (2.1g).

1H NMR (DMSO-d ₆ ) δ 8.17 (s, 1H), 8.08 (d, 1H), 7.58 (d, 1H), 3.71 (m, 4H), 2.99 (m, 4H).

Step 2: 4-Μθ ^1ϊηο-3-(ΜΑυοΓθπΐ6ΐ1^1^6ηζοϊϋ acid

4-Morpholino-3-(trifiuoromethyl)benzonitrile (2.1g, 8.016mmol) was dissolved in a mixture of tetrahydrofurane/methanol/water (1:1:1, v/v/v, 20mL), sodium hydroxide (1.9g, 48.10mmol) was added thereto and stirred for 40 hours with reflux. The reaction mixture was cooled to room temperature and IN hydrochloride was added slowly thereto to pH 3-4. The resulting solid was filtered, washed with water and dimethyl ether and then dried to obtain the title compound (694mg).

1H NMR (300MHz, DMSO-d ₆ ) δ 8.15 (d, 2H), 7.57 (d, 1H), 3.70 (m, 4H), 2.95 (m, 4H). Step 3 : -(4-methyl-3-mtrophenyl)-4-mo holino-3-(trifluoromethyl)benzamide

4-Morpholino-3-(trifluoromethyl)benzoic acid (694mg, 2.523mmol) was dissolved in dimethylformamide (5.6mL), HATU(1.92g, 5.047mmol) and N,N- diisopropylethylamine (1.37mL, 7.851mmol) were added thereto and stirred at room temperature for 30 minutes. 4-Methyl-3-nitroaniline (256mg, 1.682mmol) was added to the reaction mixture and subjected to a reaction at 45°C for 19 hours. After cooling to room temperature the reaction mixture was diluted with ethyl acetate (lOmL) and washed with an aqueous sodium hydrogen carbonate solution (lOmL). The organic layer was washed with water, dried with sodium sulfate, and the solvent was distilled under reduced pressure. The concentrated mixture was crystallized using ethyl acetate (3mL) n-hexane (30mL) to obtain the title compound (673mg) as a brown solid.

1H NMR (300 MHz, DMSO-d ₆ ) δ = 10.7 (s, 1H), 8.50 (s, 1H), 8.25 (m, 2H), 7.98 (d, 1H), 7.65 (d, 1H), 7.49 (d, 1H), 3.72 (m, 4H), 2.96 (m, 4H).

Step 4 : N-(3 -amino-4-methylphenyl)-4-morpholino-3 -(trifluoromethyl)benzamide

N-(4-memyl-3-nitrophenyl)-4-morpholino-3-(trifluoromethyl)be nzoamide (68mg, 0.17mmol) was dissolved in ethanol (1.7mL), tin chloride (190mg, 0.83mmol) was added thereto, and the mixture was subjected to a reaction with reflux for 3 hours. The reaction mixture was cooled to room temperature, diluted with ethyl acetate and then washed with an aqueous sodium hydrogen carbonate solution. The organic layer was washed with distilled water (lOmL x 3), dried with sodium sulfate, and the solvent was distilled under reduced pressure. The concentrated solution was crystallized using ethyl acetate (2mL) and n-hexane (20mL) to obtain the title compound as a bright yellow solid (26mg).

1H NMR (300 MHz, DMSO-d ₆ ) δ 10.0 (s, 1H), 8.21 (m, 2H), 7.61 (d, 1H), 7.07 (s, 1H), 6.87 (d, 1H), 6.81 (d, 1H), 4.84 (s, 2H), 3.72 (m, 4H), 2.94 (m, 4H), 2.04 (s, 3H).

Example 40: 4-(Cyclopropylamino)-N-(2-methyl-5-(4-morpholino-3-

(trifluorometh l)benzamido)phenyl)thieno[3,2-d]pyrimidme-7-carboxamide

4-(Cyclopropylamino)thieno[3 ,2-d]pyrimidine-7-carboxylic acid (23mg, O.lOmmol) was dissolved in dimethylformamide l.OmL, HATU (76mg, 0.20mmol) and N,N-diisopropylethylamine (0.08 lmL, 0.47mmol) was added thereto, and the mixture was stirred at room temperature for 30 minutes. N-(3-amino-4-methylphenyl)-4-morpholino- 3-(trifluoromethyl)benzoamide (25mg, 0.066mmol) was added to the reaction mixture and subjected to a reaction at 45°C for 17 hours. After cooling to room temperature the reaction mixture was diluted with ethyl acetate (5mL) and washed with an aqueous sodium hydrogen carbonate solution (5mL). The organic layer was washed with brine (5mL x 3), dried with sodium sulfate, and the solvent was distilled under reduced pressure. The concentrated reactant was crystallized using ethyl acetate (lmL) and n-hexane (lOmL) to obtain the title compound as a bright brown solid (34mg).

Ή NMR (300 MHz, DMSO-d ₆ ) δ 11.8 (s, 1H), 10.4 (s, 1H), 8.95 (s, 1H), 8.66 (d, 2H), 8.50 (s, 1H), 8.26 (d, 2H), 7.65 (d, 1H), 7.50 (d, 1H), 7.27 (d, 1H), 3.74 (m, 4H), 3.03 (m, 1H), 2.98 (m, 4H), 2.48 (s, 3H), 0.85 (m, 2H), 0.68 (m, 2H).

The compound of Preparation Example 9 was prepared according to Reaction Scheme 16 below:

Reaction Scheme 16

Preparation Example 9: t-Butyl 3-amino-4-methylphenyIcarbamate

Step 1 : t-But l 4-methyl-3-nitrophenylcarbamate

4-Methyl-3-nitroaniline (5.0g, 32.86mmol) was dissolved in anhydrous THF (25mL). t-Butyl bicarbonate (7.5mL, 32.86mmol) dissolved in anhydrous THF (25mL) was added slowly to the reaction mixture, which was stirred at 65°C for 18 hours. After the reaction was completed the remaining organic solvent was distilled under reduced pressure, and water was added thereto. The resulting yellow solid was filtered and dried to obtain the title compound without further purification. Ή NMR (400 MHz, DMSO-d ₆ ) 69.76 (s, 1H), 8.23 (d, 1H), 7.57 (dd, 1H), 7.39 (d, 1H), 2.43 (s, 3H), 1.48 (s, 9H).

Step 2: t-Butyl 3-amino-4-methylphenylcarbamate

t-Butyl 4-methyl-3-nitrophenyl carbamate (7.6g, 30.13mmol) was dissolved in ethanol (167mL) and subjected to a reaction under 50 psi H ₂ gas. The reaction mixture was stirred for 2 hours and then concentrated by filtering with Celite to obtain the title compound without further purification.

Ή NMR (400 MHz, DMSO-d ₆ ) 68.92 (s, 1H), 6.82 (s, 1H), 6.64 (d, 1H), 6.48 (d, 1H), 4.73 (s, 2H), 1.88 (s, 3H), 1.44 (s, 9H).

The compound of Example 82 was prepared according to Reaction Scheme 17 below:

Example 82 : 4-(Cyclopropylamino)-N-(2-methyl-5-(4-(4-methyl- 1 H-imidazole- l-yl)-3 (trifluoromethyl)benzoamido)phenyl)thieno[3,2-d]pyrimidine-7 -carboxamide

Step 1: t-Butyl 3-(4-(cyclopropylamino)thieno[3,2-d]pyrimidine-7-carboxamido )-4 methylphenylcarbamate

4-(Cyclopropylamino)thieno[3,2-d]pyrimidine-7-carboxylic acid (5.3g, 22.49mmol) obtained in Step 3 of Preparation Example 4 was dissolved in DMF (60mL). t-Butyl 3-amino-4-methylphenylcarbamate (6g, 26.99mmol) obtained in Step 2 of Preparation Example 9, EDCI (6.5g, 33.74mmol), HOBT (6.1g, 44.98mmol) and TEA (4.7mL, 33.74mmol) were sequentially added to the reaction mixture, which was stirred at room temperature for 24 hours. After the reaction was completed water (120mL) was added thereto, and the resulting yellow solid was filtered and dried to obtain the title compound without further purification.

MS m/z [M+l] 439.84.

Step 2: N-(5-amino-2-methylphenyl)-4-(cyclopropylamino)thieno[3,2-d] pyrimidine-7- carboxamide

t-Butyl 3 -(4-(cyclopropylamino)thieno[3 ,2-d]pyrimidine-7-carboxamido)-4- methylphenylcarbamate (7g, 15.93mmol) was dissolved in DCM (80mL), TFA (12mL, 159.3mmol) was added thereto, and the mixture was stirred at room temperature for 8 hours. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to remove the organic solvent and the remaining TFA, and water (20mL) was added thereto. The reaction mixture was neutralized with 2.0 N NaOH solution, and the resulting yellow solid was filtered and dried to obtain the title compound without further purification.

MS m/z [M+l] 339.87

Step 3 : 4-(Cyclopropylamino)-N-(2-methyl-5-(4-(4-methyl- 1 H-imidazole- 1 -yl)-3- (trifluoromethyl)benzoamido)phenyl)thieno[3,2-d]pyrimidine-7 -carboxamide

The procedure of Step 5 of Example 1 was repeated except for using N-(5-amino-2- methylphenyl)-4-(cyclopropylamino)thieno[3,2-d]pyrimidine-7- carboxamide and 4-(4- memyl-lH-imidazole-l-yl)-3-(trifluoromethyl)benzoic acid to obtain the title compound.

Ή NMR (300 MHz, DMSO-d ₆ ) δ 11.8 (s, 1H), 10.6 (s, 1H), 8.93 (s, 1H), 8.67 (d, 2H), 8.47 (s, 2H), 8.39 (d, 1H), 7.94 (s, 1H), 7.75 (d, 1H), 7.60 (m, 1H), 7.29 (m, 1H), 7.11 (s, 1H), 3.05 (m, 1H), 2.18 (s, 3H), 0.84 (m, 2H), 0.66 (m, 2H)

Examples 41 to 81 and Examples 83 to 121

The procedure of Example 39, 40 or 82 was repeated except for using an appropriate starting compound (see Table 1).

Example 122: 4-Amino-N-(5-(3-(4-chloro-3-(trifluoromethyl)plienyl)ureido) -2- methylphenyl)thieno [3,2-d] pyrimidine-7-carboxamide

4-Amino-N-(5-amino-2-methylphenyl)thieno[3,2-d]pyrimidine -7-carboxamide (50mg, 0.167mmol) obtained in Step 3 of Example 39 was dissolved in anhydrous THF (ImL), TEA (47μί, 0.334mmol) and triphosgene (17mg, 0.059mmol) were added thereto at 0°C, and the mixture was stirred for 2 hours. 4-Chloro-3- (trifluoromethyl)benzeneamine (49mg, 0.251mmol) was added to the reaction mixture, which was stirred at room temperature for 10 hours. The reaction mixture was diluted with ethyl acetate (lOmL) and washed with brine. The organic layer was dried with MgSC«4, filtered and then concentrated under reduced pressure. The resulting mixture was purified by silica gel chromatography (MeOH/DCM=3/97) to obtain the title compound (54mg, 60%). 1H NMR (300 MHz, DMSO-d ₆ ) δ 11.70 (s, IH), 9.01 (s, IH), 8.91 (s, IH), 8.87 (s, IH), 8.56 (s, IH), 8.40 (s, IH), 8.11 (s, IH), 7.89 (br s, 2H), 7.60 (m, 2H), 7.28 (d, IH), 7.18 (d, IH), 2.40 (s, 3H). Example 123: 4-(Cyclopropylammo)-N-(2-methyl-5-(3-(3-(4-methyl-lH-imidazo le-l- yl)-5-(t fluoromethyl)phenyl)ureido)phenyl)thieno[3,2-d]pyrimidine-7- carboxamide

The procedure of Example 122 was repeated except for using the compound obtained in Step 2 of Example 82 instead of N-(5-amino-2-methylphenyl)-4- (cyclopropylamino)thieno[3,2-d]pyrimidine-7-carboxamide to obtain the title compound.

1H NMR (300 MHz, DMSO-d ₆ ) δ 11.76(br, 2H), 9.06(d, IH), 8.94(s, IH), 8.64(m, 2H), 8.51(s, IH), 8.40(d, IH), 8.21(s, IH), 7.94(d, IH), 7.53(d, IH), 7.32(m, 2H), 7.19(m, IH), 3.04(m, IH), 2.37(s, 3H), 2.18(s, 3H), 0.85(m, 2H), 0.69(m, 2H). compound of Example 124 was prepared according to Reaction Scheme 17 below:

eaction Scheme 17

Example 124 : 3-((4-(Cyclopropylamino)thieno [3,2-d] pyrimidine-7-yl)ethynyI)-4- methyI-N-(3-(4-methyI-lH-imidazole-l-yl)-5-(trifluoromethyl) phenyl)benzamide

Step 1 : -(Methylmio)-7-((trimethylsilyl)ethynyl)thieno[3,2-d]pyrimid ine

7-Bromo-4-(memylthio)thieno[3,2-d]pyrimidine (3g, 11.49mmol) was dissolved in acetonitrile (30mL) and then bis(triphenylphosphine)palladium(II) dichlonde (204mg, 0.29mmol), Cul (76mg, 0.40mmol), dicyclohexylamine (2.5mL, 12.64mmol) and ethynyltrimethylsilane (3.2mL, 22.98mmol) were added thereto. The reaction mixture was treated with N ₂ gas for 15 minutes and stirred at 80°C for 14 hours. The reaction mixture was cooled to room temperature, filtered using Celite and washed with ethyl acetate (50mL). The filtrate was washed with brine and concentrated by drying with MgS0 ₄ . The resulting mixture was purified by silica gel chromatography (ethyl acetate/hexane=l :9~2:8) to obtain the title compound (2.1g, 65% yield) as a yellowish solid.

1H NMR (400MHz, CDC1 ₃ ) δ 9.05 (s, 1H), 7.99 (s, 1H), 2.74 (s, 3H), 0.30 (s, 9H), MS m/z : 279.34 [M+l].

Step 2: 7-Em nyl-4-(methylthio)thieno[3,2-d]pyrimidine

4-(Methylthio)-7-((trimemylsilyl)ethynyl)thieno[3,2-d]pyrimi dine (500mg, 1.8mmol) was dissolved in THF (5mL) at 1.0M and then tetrabutylammonium fluoride solution (3.6mL, 3.6mmol) was added thereto. The reaction mixture was stirred at room temperature for 30 minutes and then concentrated. The resulting mixture was purified by silica gel chromatography (ethyl acetate/hexane=2:8~3:7) to obtain the title compound (340mg, 91% yield) as a yellowish solid.

1H NMR (400MHz, CDC1 ₃ ) δ 9.07 (s, 1H), 8.03 (s, 1H), 3.44(s, 1H), 2.88 (s, 3H), MS m/z : 207.23 [M+l].

Step 3: Ethyl 4-methyl-3-((4-(memylmio)thieno[3,2-d]pyrimidine-7-yl)ethyny l)benzoate

7-Ethynyl-4-(methylthio)thieno[3,2-d]pyrimidine (850mg, 4.12mmol) was dissolved in DMF (9mL) and then ethyl 3-iodo-4-methyl benzoate (902mg, 4.12mmol), Pd(PPh ₃ ) ₄ (242mg, 0.2 lmmol), Cul(59mg, 0.3 lmmol) and DIEA(1.OmL, 6.18mmol) were added thereto. The reaction mixture was stirred at room temperature for 16 hours and concentrated by filtering with Celite. The resulting mixture was purified by silica gel chromatography (ethyl acetate/hexane=l:9~2:8) to obtain the title compound(l.lg, 72% yield) as a yellowish solid.

1H NMR (400MHz, CDC1 ₃ ) δ 9.11 (s, 1H), 8.75 (s, 1H), 8.07 (s, 1H), 7.90 (d, 1H),

7.52 (d, 1H), 4.33 (q, 2H), 2.78 (s, 3H), 2.61 (s, 3H), 1.34 (t, 3H), MS m/z : 369.44 [M+l].

Step 4: Ethyl 3-((4-(cyclopropylamino)thieno[3,2-d]pyrimidine-7-yl)ethynyl )- 4-methyl benzoate

Etoyl-4-me&yl-3-((4-(methyltoio)tm ^' e

(440mg, 1.19mmol) was dissolved in methylene chloride (6mL) and then m-CPBA (616mg, 3.57mmol) was added thereto at 0°C. The reaction mixture was stirred at 0°C for 30 minutes and methylene chloride (20mL) was added thereto. The reaction mixture was washed with a sat. aqueous NaHC0 ₃ solution (30mL x 3). The organic layer was dried with MgS0 ₄ , filtered with Celite and concentrated to obtain the title compound without further purification.

1H NMR (400MHz, CDC1 ₃ ) δ 9.58 (s, IH), 9.13 (s, IH), 8.10 (s, IH), 7.94 (d, IH), 7.70 (d, IH), 4.33 (q, 2H), 3.54 (s, 3H), 2.63 (s, 3H)), 1.34 (t, 3H), MS m/z : 385.50 [M+1].

Step 5: Emyl-3-((4-(cyclopropylamino)thieno[3,2-d]pyrimidine-7-yl)et hynyl)- 4-methyl benzoate

Ethyl-3-((4-(cyclopropylamino)thieno[3,2-d]pyrimidine-7-yl)e thynyl)-4-methyl benzoate (50mg, 0.13mmol) was dissolved in dioxane (lmL) and then cyclopropylamine (37mg, 0.65mmol) and DIEA (O.lmL, 0.65mmol) were added thereto. The reaction mixture was stirred at 120°C for 8 hours and cooled to room temperature. Ethyl acetate (5mL) was added thereto and the reaction mixture was washed with brine. The organic layer was dried with MgS0 ₄ , filtered with Celite and concentrated. The resulting mixture was purified by silica gel chromatography (ethyl acetate/hexane=2:8~4:6) to obtain the title compound (32mg, 65% yield).

1H NMR (400MHz, DMSO-<¾ δ 8.52 (s, IH), 8.47 (s, IH), 8.09 (m, IH), 8.03 (s, IH), 7.89(dd, IH), 7.51 (d, IH), 4.32 (q, 2H), 3.58 (m, 4H), 2.58 (s, 3H), 1.32 (t, 3H), MS m/z : 382.59 [M+1].

Step 6:

3-((4-(Cyclopropylamino)thieno[3,2-d]pyrimidine-7-yl)ethynyl )-4-methylbenzoic acid

The procedure of Step 4 of Example 1 was repeated except for using the compound obtained in Step 5 of Exmaple 123 as a starting material to obtain the title compound.

MS m/z : 350.09 [M+l].

Step 7: 3-((4-(Cyclopropylamino)tmeno[3,2-d]pyrimidine-7-yl)ethynyl) -4-methyl-N-(3-(4- methyl- 1 H-imidazole- 1 -yl)-5-(trifluoromethyl)phenyl)benzamide

The procedure of Step 5 of Example 1 was repeated except for using the compound obtained in Step 6 in Example 124 as a starting material to obtain the title compound.

MS m/z : 573.17 [M+l].

Example 125 : N-(3-((4-(cyclopropylamino)thieno [3,2-d] pyrimidine-7-yl)ethynyl)-4- methylphenyl)-3-(4-methyI-lH-imidazole-l-yl)-5-(trifluoromet hyl)benzamide

The procedure of Step 3 of Example 124 was repeated except for using 2-iodo-l- methyl-4-nitrobenzene (TCI Laboratory Chemicals, Cat. # 10706, CAS [7745-92-8]) instead of 3-iodo-4-methyl benzoate to obtain the product and then the procedure of Steps 1 to 3 of Example 4 was repeated using the product to obtain the title compound.

MS m/z : 573.19 [M+l]. Example 126 : 3-((4-(Cyclopropylamino)thieno [3,2-d] pyrimidine-7-yl)ethynyl)-4- methyl- -(4-((4-methylpiperazme-l-yl)methyl)-3-(trifluoromethyl)phen yl)benzainide

The procedure of Step 5 of Example 1 was repeated except for using 3-((4-

(cyclopropylamino)thieno[3 ,2-d]pyrimidine-7-yl)ethynyl)-4-methylbenzoic acid obtained in Step 6 of Example 124 and (4-((4-methylpiperazine-l-yl)methyl)-3- (trifluoromethyl)amline to obtain the title compound.

MS m/z : 605.15 [M+l].

Example 127: 3-((4-(Cyclopropylanm o)thieno[3,2-d]pyriinidine-7-yl)ethynyl)-N-(4- ((4-ethylpiperazme-l-yl)methyl)-3-(trifluoromethyl)phenyl)-4 -methylbenzamide

The procedure of Step 5 of Example 1 was repeated except for using 3-((4- (cyclopropylamino)tWeno[3,2-d]pyrimidine-7-yl)ethynyl)-4-met hylbenzoic acid obtained in Step 6 of Example 124 and (4-((4-ethylpiperazine-l-yl)methyl)-3- (trifluoromethyl)aniline to obtain the title compound.

1H NMR (DMSO, 300MHz) δ 10.54(s, IH), 8.54(s, IH), 8.48(s, IH), 8.19(m, 3H), 8.05(d, IH), 7.91(d, IH), 7.70(d, IH), 7.51(d, IH), 3.55(s, 2H), 2.98(m, IH), 2.59(s, 3H), 2.38(m, 10H), 0.97(t, 3H), 0.80(m, 2H), 0.64(m, 2H).

Example 128: 3-((4-(Cyclopropylammo)thieno[3,2-d]pyrimidine-7-yl)ethynyl) -4- methyl-N-(4-(l-methylpiperazme-l-yloxy)-3-(trifluoromethyl)p henyl)benzamide

The procedure of Step 5 of Example 1 was repeated except for using 3-((4- (cyclopropylamino)thieno[3,2-d]pyrimidine-7-yl)ethynyl)-4-me thylbenzoic acid obtained in Step 6 of Example 124 and 4-(l -methylpiperidine-4-yloxy)-3-(trifluoromethyl)aniline to obtain the title compound.

1H NMR (DMSO, 300MHz) δ 10.41(s, IH), 8.56(s, IH), 8.49(s, IH), 8.24(d, IH), 8.18(d, IH), 8.1 l(d, IH), 8.03(d, IH), 7.92(d, IH), 7.52(d, IH), 7.33(d, IH), 4.60(m, IH), 3.01(m, IH), 2.60(s, 3H), 2.38(m, 4H), 2.28(s, 3H), 1.93(m, 2H), 1.71(m, 2H), 0.83(m, 2H), 0.69(m, 2H).

The structures, NMR and MS m/z data of the compounds obtained in Examples 1 to 128 are summarized in Table 1 .

Table 1

1 H-imidazole- 1 -yl)-5 -(trifluoromethyl)benzamide

carboxamide Ή NMR (400MHz, DMSO- d6)811.86 (s, 1H), 10.20 (s, 1H), 8.97 (s, 1H), 8.85-8.80 (m, 1H), 8.60 (s, 1H), 7.95 (br, 2H), 7.67- 7.64 (m, 1H), 7.46-7.43 (m, 1H), 7.11-7.10 (m, 2H), 6.30-6.20 (m, 1H), 3.74 (s, 6H), 2.54 (s, 3H),

4-amino-N-(5 -(3 ,5 -dimethoxyphenylcarbamoyl)-2-methylphenyl)thieno[3 ,2- d]pyrimidine-7-carboxamide

Ή NMR (400 MHz, DMSO-de) δ 11.85 (s, 1H), 10.56 (s, 1H), 10.06 (s, 1H), 9.00 (s, 1H), 8.87 (s, 1H),

1 I S I 8.73 (s, 1H), 8.26 (s, 1H), 8.08 (d,

1H), 7.72 (d, 1H), 7.63-7.57 (m, 3H), 7.47 (d, 2H), 7.01 (d, 2H), 3.79 (s, 3H), 2.54 (s, 3H); MS m/z [M+l] 578.07.

4-(4-methoxyphenylamino)-N-(2-methyl-5 -(3 -

(trifluoromethyl)phenylcarbamoyl)phenyl)tlueno[3,2-d]pyri midine-7-carboxamide

Ή NMR (400 MHz, DMSO-d _f i) δ 11.90 (s, 1H), 10.57 (s, 1H), 10.12 (s, 1H), 9.04 (s, 1H), 8.88 (s, 1H), 8.81 (s, 1H), 8.26 (s, 1H), 8.08 (d, 1H), 7.72 (d, 1H), 7.61 (t, 1H), 7.48 (dd, 2H), 7.15 (s, 2H), 3.80 (s, 6H), 3.69 (s, 3H), 2.56 (s, 3H); MS m/z [M+l] 638.08

N-(2-methyl-5-(3-(trifluoromethyl)phenylcarbamoyl)phenyl) -4-(3,4,5- trimethoxyphenylamino)thieno[3,2-d]pyrimidine-7-carboxamide

Ή NMR (400 MHz, DMSO-de) δ 11.74 (s, 1H), 10.52 (s, 1H), 10.45 (s, 1H), 9.12 (s, 1H), 8.98 (br s, 1H), 8.87 (d, 2H), 8.30 -8.26 (m, 2H), 8.08 (d, 1H), 7.73 (d, 1H), 7.61 (t, 1H), 7.53-7.45 (m, 3H), 2.56 (br s, 6H); MS m/z [M+l] 563.06

N-(2-methyl-5-(3-(trifluoromethyl)phenylcarbamoyl)phenyl)-4- (6-methylpyridine-3- ylamino)thieno [3 ,2-d]pyrimidine-7-carboxamide

methylbenzamido)-2-(trifluoromethyl)benzyl acetate (s,

carboxamide

4-(cyclopropylamino)-N-(5-(3-(3-(dimethylamino)propylamino)- 5-

7-carboxamide Ή NMR (300 MHz, DMSO- d6)51 1.39 (s, 1 H), 10.56 (s, 1 H), 8.93 (s, 1 H), 8.77 (m, 2 H), 8.69 (d, 1 H), 8.58 (s, 1 H), 7.86 (m, 2 H), 7.57 (dd, 1 H), 7.25 (d, 1 H), 2.43 (s, 3 H)

4-amino-N-(5-(isonicotniamido)-2-methylphenyl)tlneno[3,2-d]p yrin^

carboxamide

•H NMR (300 MHz, DMSO- de)511.75 (s, 1 H), 10.55 (s, 1 H), 8.92 (s, 1 H), 8.68 (d, 1 H), 8.59 (s, 1 H), 8.35 (d, 2 H), 8.12 (s, 1 H), 7.92 (br s, 2 H), 7.59 (dd, 1 H), 7.50 (s, 1 H), 7.28 (d, 1 H), 6.98 (s, cv 1 H), 2.44 (s, 3 H), 2.36 (s, 3 H);

MS m/z [M+l] 551.88

4-amino-N-(2-methyl-5-(3 -(2-methyl- 1 H-imidazole- 1 -yl)-5- (trifluoromethyl)benzamido)phenyl)tmeno[3,2-d]pyrimidine-7-c arboxamide

Ή NMR (300 MHz, DMSO- d6)511.71 (s, 1 H), 10.33 (s, 1 H), 8.92 (s, 1 H), 8.66 (d, 1 H), 8.57 (s, 1 H), 7.93 (m, 2 H), 7.81 (m, 2 H), 7.57 (m, 2 H), 7.43 (br t, 1 H), 7.25 (d, 1 H), 2.43 (s, 3 H)

4-annno-N-(5-(3-fluorophenylamido)-2-memylphenyl)thieno[3,2- d]pyrimidine-7- carboxamide

¾ NMR (300 MHz, DMSO- d6)511.71 (s, 1 H), 10.53 (s, 1 H), 8.91 (s, 1 H), 8.59 (m, 2 H), 7.97 (m, 2 H), 7.83 (d, 1 H), 7.77 (d, 1 H), 7.69 (br t, 2 H), 7.47 (d, 1 H), 7.24 (d, 1 H), 2.45 (s, 3 H)

4-amino-N-(2-memyl-5-(2-(trifluoromethyl)benzamido)phenyl )tWeno[3,2-d]pyrimidine- 7-carboxamide

cyclopropylisoxazol-3-carboxamide

d]pyrimidine-7-carboxamide carboxamide

yl)phenylamino)thieno[3,2-d]pyrimidine-7-carboxamide

(cyclopropylamino)thieno[3,2-d]pyrimidine-7-carboxamide (S)-4-(cyclopropylamino)-N-(5-(4-(3-(dimethylamino)pyrrolidi ne-l-yl)-3- (trifluoromethyl)benzoamido)-2-methylphenyl)tWeno[3,2-d]pyri midine-7-carboxamide

1H NMR (DMSO, 300MHz); δ 1 1.74(s, 1H), 10.52(s, 1H), 8.89(s, 1H), 8.64(m, 2H), 8.45(s, 1H), 8.39(s, 1H), 8.19(m, 3H), 7.70(s, 1H), 7.58(d, 1H), 7.28(d, 1H), 4.10(m, 1H), 2.80(m, 2H), 2.43(s, 3H), 2.17(d, 6H), 1.96(m, 4H), 1.65(m, 2H).

N-(2-methyl-5-(3-(4-methyl-lH-imidazole-l-yl)-5-

(trifluoromethyl)benzoamido)phenyl)^4-( 1 -methylpiperidine-4-ylamino)tbieno [3,2- d]pyrimidine-7-carboxamide

1H NMR (DMSO, 300MHz); δ 11.75(s, 1H), 10.41(s, 1H), 8.93(s, 1H), 8.64(s, 2H), 8.49(s, 1H), 8.24(s, 1H), 8.21(d, 1H), 7.63(d, 1H), 7.54(dd, 1H), 7.24(d, 1H), 3.30(m, 1H), 3.02(q, 4H), 2.43(s, 3H), 0.94(t, 6H), 0.82(m, 2H), 0.68(m, 2H).

4-(cyclopropylamino)-N-(5-(4-(diethylamino)-3-(trifluorometh yl)benzoamido)-2- methylphenyl)tMeno[3,2-d]pyrimidine-7-carboxamide

1H NMR (DMSO, 300MHz); δ 11.72(s, 1H), 10.1 l(s, 1H), 8.92(s,

_U «N O O 1H), 8.64(s, 1H), 8.59(s, 1H),

8.49(s, 1H), 8.12(s, 1H), 8.06(d, 1H), 7.53(d, 1H), 7.22(d, 1H), 7.00(d, 1H), 5.14(d, 1H), 3.30(m,

Λ 1H), 3.03(m, 1H), 2.68(m, 2H),

XT 2.41(s, 3H), 2.24(s, 3H), 2.06(m,

2H), 1.90(m, 2H), 1.60(m, 2H), 0.83(m, 2H), 0.67(m, 2H).

4-(cyclopropylamino)-N-(2-methyl-5 -(4-( 1 -methylpiperidine-4-ylamino)-3 - (trifluoromethyl)benzoamido)phenyl)tlue^^

Ή NMR (300 MHz, DMSO- d6)512.21(s, 1H), 10.72(s, 1H), 9.00(d, 2H), 8.65(s, 1H), 8.50(s, 1H), 8.46(s, 1H), 8.40(s, 1H), 8.24(s, 1H), 8.17(s, 1H), 7.71(m, 2H), 7.57(d, 1H), 3.04(m, 1H), 2.18(s, 3H), 0.83(m, 2H), 0.68(m, 2H).

N-(2-chloro-5-(3-(4-methyl-lH-imidazole-l-yl)-5-

(trifluoromethyl)berizoanndo)phenyl)^-(phenylarmno)tWeno[3,2 -d]pyrimid

(trifluoromethyl)benzoamido)phenyl)tMeno[3,2-d]pyrimidine-7- carboxamide

methylphenyl)thieno[3,2-d]pyrimidine-7-carboxamide

(trifluoromethyl)benzoamido)phenyl)tbjeno[3,2-d]pyrmiidine-7 -carboxamide Ή NMR (300 MHz, DMSO- d6)511.62(br, 1H), 10.58(br, 1H), 9.20(d, 2H), 8.72(d, 1H), 8.48(s, 1H), 8.41(s, 1H), 8.24(s, 1H), 8.20(s, 1H), 8.13(d, 2H), 7.73(s,

120

1H), 7.61(m, 4H), 7.34(d, 1H), r* 2.49(s, 3H), 2.20(s, 3H)

4-benzoamido-N-(2-methyl-5-(3-(4-methyl-lH-imidazole-l-yl)-5 - (trifluoromethyl)benzoamido)phenyl)thieno[3,2^]pyrimidine-7- carboxamide

Ή NMR (300 MHz, DMSO- d6)512.21(br, 1H), 10.64(br, 1H), 8.98(s, 1H), 8.89(dd, 1H), 8.70(s, 1H), 8.5 l(s, 1H), 8.46(s, 1H), 8.40(s, 1H), 8.24(s, 1H), 8.17(s,

121 1H), 7.72(s, 1H), 7.70(m, 1H),

λ ^Ν 7.37(dd, 1H), 3.14(m, 1H), 2.18(s,

3H), 0.84(m, 2H), 0.68(m, 2H).

4-(cyclopropylamino)-N-(2-fluoro-5-(3-(4-methyl-lH-imidaz ole-l-yl)-5- (trifluoromethyl)benzoanndo)phenyl)tbieno[3,2-d]pyrimidine-7 -carboxamide

Ή NMR (300 MHz, DMSO- 4)811.70 (s, 1H), 9.01 (s, 1H), 8.91 (s, 1H), 8.87 (s, 1H), 8.56 (s, 1H), 8.40 (s, 1H), 8.11 (s, 1H), 7.89

122 Η (br s, 2H), 7.60 (m, 2H), 7.28 (d,

^Η CF ₃ 1H), 7.18 (d, 1H), 2.40 (s, 3H)

4-amino-N-(5-(3-(4-chloro-3-(trifluoromethyl)phenyl)ureid o)-2- methylphenyl)tbieno[3,2-d]pyrimidine-7-carboxamide

Ή NMR (300 MHz, DMSO-

CF ₃ d6)511.76(br, 2H), 9.06(d, 1H),

8.94(s, 1H), 8.64(m, 2H), 8.51(s, 1H), 8.40(d, 1H), 8.21(s, 1H), 7.94(d, 1H), 7.53(d, 1H), 7.32(m,

123 2H), 7.19(m, 1H), 3.04(m, 1H),

2.37(s, 3H), 2.18(s, 3H), 0.85(m, 2H), 0.69(m, 2H).

4-(cyclopropylamino)-N-(2-methyl-5-(3-(3-(4-methyl-lH-imidaz ole-l-yl)-5- (trifluoromethyl)phenyl)ureido)phenyl)thieno[3,2-d]pyrimidin e-7-carboxamide

ethylpiperazine- 1 -yl)methyl)-3 -(trifluoromethyl)phenyl)-4-methylbenzamide 1H NMR (DMSO, 300MHz); δ

10.41(s, 1H), 8.56(s, 1H), 8.49(s, 1H), 8.24(d, 1H), 8.18(d, 1H), 8.1 l(d, 1H), 8.03(d, 1H), 7.92(d, 1H), 7.52(d, 1H), 7.33(d, 1H), 4.60(m, 1H), 3.01(m, 1H), 2.60(s,

128

Y 3H), 2.38(m, 4H), 2.28(s, 3H),

1.93(m, 2H), 1.71(m, 2H), 0.83(m, 2H), 0.69(m, 2H)

3-((4-(cyclopropylamino)thieno[3,2-d]pyrimidine-7-yl)ethynyl )^-methyl-N-(4-(l- methylpiper^ine-l-yloxy)-3-(trifluoromethyl)phenyl)benzamide

Test 1: Kinase activation assay (IC ₅ o) A kinase domain (Millipore) comprising the entire cytoplasmic sequences of Bcr-

Abl* ¹ and Bcr-Abl™ ⁵¹ mutation kinases and a tyrosine kinase assay kit (Panvera, P2837) were used to measure an inhibitory activation on Bcr-abl. Other kinases such as FGFR, Fit, KDR, PDGFR, Fms, Kit, Raf, Tie2, Src, and Ret were also measured by the same method. In order to prepare a sample, the each compound obtained in Examples 1 to 128 was diluted 10 times with a kinase buffer to a concentration of 10-0.0001 μΜ.

The each kinase was dilluted with a kinase buffer to a concentration of 30~300ng/assay by using the each Kd value. The kinase buffer consisted of lOOmM HEPES (PH 7.4), 25mM MgCl ₂ , lOmM MnCl ₂ , and 250μΜ Na ₃ V0 ₄ , being kept refrigerated after filtered.

The test was conducted in 96 well polystyrene round bottom plates. First, ΙΟμΙ. of the diluted sample was combined with ΙΟμΙ, of the enzyme solution at room temperature for 10 minutes using a shaker. ΙΟμΙ, of Poly (Glu, Tyr) 4: 1 (Sigma) matrix solution in an appropriate concentration and ΙΟμί, of ATP solution (5~300μΜ) were added to the sample and subjected to a reaction at room temperature for 60 minutes in a shaker. After 60 minutes, an EDTA solution was added thereto to complete the kinase reaction and the reaction mixture was left in a shaker for 5 minutes at room temperature. A tracer, an antibody, and a fluorescence polarization (FP) diluted reaction solution were mixed at a ratio of 1 : 1 :3 to prepare a detecting solution, 50μί, of which was added to the each sample and subjected to a reaction at room temperature for 30 minutes in a shaker. As fluorescing, light was blocked out and the FP value was measured using a FP instrument (excitation wavelength: 485nm, and emission wavelength: 530nm).

And the inhibition on kinase reaction was calculated for the sample in the range of 0-100% as compared to the control group, and the concentration on x-axis at 50% inhibition was obtained as an inhibitory concentration (IC ₅₀ ) value.

50% inhibitory concentration (IC50)

Y = bottom + (top-bottom) / (1 + 10exp(X-logIC ₅₀ ))

The IC50 values of Bcr-Abl ^wt and Bcr-AbI ^T3151 kinases for the compounds obtained in the examples were summarized in Table 2.

Table 2

As shown in Table 2, the inventive compounds have excellent inhibitory activity on Bcr-Abl" ¹ and Bcr-Abl ^T3151 kinases.