KR20050091462A - Furopyrimidine compound and ddr2 tyrosine kinase activity inhibitor comprising the same - Google Patents

Abstract

The present invention relates to novel furopyrimidine compounds, their pharmaceutically acceptable salts, and inhibitors of DDR2 (Discoidin Domain Receptor 2) tyrosine kinase activity comprising them. The puropyrimidine compound may be a compound defined by Chemical Formula 1, Chemical Formula 2 or Chemical Formula 3, precursors thereof, or exist in free form or in acid addition salt form. Since the puropyrimidine compound of the present invention has an effect of inhibiting the activity of DDR2 tyrosine kinase, it can be used to treat diseases involving DDR2 tyrosine kinase activity, such as cirrhosis, rheumatoid arthritis, cancer and the like.

[Formula 1]

[Formula 2]

[Formula 3]

Description

FUROPYRIMIDINE COMPOUND AND DDR2 TYROSINE KINASE ACTIVITY INHIBITOR COMPRISING THE SAME}

The present invention relates to novel furopyrimidine compounds, pharmaceutically acceptable salts thereof, and DDR2 (Discoidin Domain Receptor 2) tyrosine kinase activity inhibitors comprising them, the compounds of the present invention inhibit the activity of DDR2 tyrosine kinase Because of its effect, it can be used to treat diseases involving DDR2 tyrosine kinase activity, such as cirrhosis, rheumatoid arthritis, cancer and the like.

In general, cognition through tyrosine kinases, which are receptors on cell membranes, is one of the ways in which cells recognize external stimuli. Receptor tyrosine kinases consist of an extracellular part exposed outside the cell, an intracellular part exposed to the intracellular cytosol, and a membrane passing part passing through the plasma membrane located in between. The extracellular portion of the receptor is the portion to which a specific ligand binds, and the intracellular portion performs the function of transmitting the activity signal of the receptor activated by the ligand into the cell.

The tyrosine kinase receptor has a domain having tyrosine kinase activity at the C-terminal site exposed in the cell. When a specific ligand is attached to the extracellular part, the kinase enzyme of the C-terminal tyrosine kinase domain is exposed to the cytoplasmic part of the receptor protein. Is activated to phosphorylate tyrosine at the C-terminus of each other on the duplex. This tyrosine phosphorylation is the most important process for transmitting signals for extracellular stimulation into cells. Many receptors have tyrosine kinase activity that transfer extracellular stimuli into cells with this mechanism. Representative examples include EGFR, PDGFR, IR, IGFR, c-fms, VEGFR and the like.

DDRid domain receptor (DDR) receptor proteins are also one of the receptors having such tyrosine kinase activity. DDR is so named because it has a similarity to discidine, a protein whose extracellular site is attached to lectins found in microorganisms. In the case of animals such as humans, it is known that DDR1 and DDR2 types exist as proteins having similarities in amino acid sequences, respectively, encoded by different genes. In humans, the presence of the DDR protein gene was known in the early 1990s, but studies of the kinase function of the protein began to attract attention when it first became known in 1997 that the ligand attached to it was collagen fiber. have.

Generally, overexpression of specific tyrosine kinase receptors due to genetic alteration or environmental influences, or structural changes may result in sustained activity regardless of the presence or absence of the ligand, or due to overproduction of the ligand, impairing its regulation of activity. When the activity is greatly increased or vice versa, the activity decreases, causing various disease phenomena.

For example, it has been found that excess activity such as EGFR or PDGFR is one of the important factors of cancer, and increased activity of VEGFR is known to be deeply involved in cancer metastasis and malignancy. Recently, several pharmaceutical companies have developed EGFR specific inhibitor compounds or VEGFR specific inhibitor compounds and developed them as anticancer drugs, or are in clinical trials. These EGFR and VEGFR specific inhibitors are both compounds that inhibit the kinase activity at the C-terminus of these receptor proteins, and in particular competitively attach ATP to the ATP attachment sites of their kinase enzymes, thereby binding the enzyme to ATP. Inhibits enzyme activity.

DDR2 protein, on the other hand, is one of the receptor tyrosine kinases and has a tyrosine kinase domain at the site exposed to cytosol. It has been demonstrated that the ideality of DDR2 protein is related to liver sclerosis and rheumatism, which are refractory human diseases.

Liver cirrhosis is a lesion that causes disease as the collagen fiber is overproduced and accumulates in the liver. Recently, experimental results have been reported to increase the expression of DDR2 protein and its kinase activity during the activation of hepatic stellate cells in liver tissue as a major cause of cirrhosis. That is, the DDR2 protein has a tyrosine kinase domain at the site exposed to the cytosol, and it is known that tyrosine kinase activity is essential for the proliferation of hepatic stellate cells of the liver tissue due to the increased activity of the DDR2 protein. This means that increased expression of DDR2 protein plays an important role in the activation of hepatic stellate cells and is a major pathogenesis factor of liver hardening. In addition, the accumulation of collagen protein, which is a ligand of DDR2 protein due to liver hardening, and the sustained activation of DDR2 protein, thereby significantly explaining that the activity of DDR2 protein kinase is a facilitating factor for liver hardening.

Another disease associated with collagen is rheumatism. Rheumatism is a disease in which immune cells are constantly activated in the cartilage area and cytokines such as TNF-α are increased, and the activity of collagen degrading enzyme MMP-1 is increased so that cartilage tissue is greatly destroyed. to be. The cells in the cartilage tissue that mainly express the MMP-1 protein are synovial fibroblasts present in the synovial membrane surrounding the cartilage.

In general, these synovial fibroblasts are well controlled in proliferation and activity under normal circumstances. Recently, however, it has been reported that MMP-1 gene expression is increased by increasing activity of DDR2 protein. In addition, expression of DDR2 protein was observed in synovial fibroblasts of cartilage tissue of rheumatoid patients. This fact indicates that increased kinase activity of DDR2 protein is the main cause of rheumatic lesions. Based on the above facts, it is inferred that a substance that specifically inhibits the kinase activity of DDR2 protein can be usefully used as a therapeutic agent for liver hardening or rheumatism.

MMP-1 and MMP-2 are known as proteins whose expression is increased by activation of kinase of DDR2 protein. These MMP proteins are known to play an important role in metastasis of cancer cells. Recently, it has been reported that the expression of DDR2 protein is characteristically increased in metastatic cancer. In other words, excessive activation of intracellular DDR2 protein may contribute to cancer malignancy.

In conclusion, the main developmental causes of lesions such as cirrhosis and rheumatism are non-ideal growth of fibroblasts in the lesion site, and the growth and growth of these fibroblast-like cells requires the expression and activation of DDR2 protein, in which case the kinase of DDR2 protein From the fact that activity is central, it can be seen that the tyrosine kinase activity of DDR2 protein is a therapeutic target for diseases caused by increased DDR2 activity such as cirrhosis, rheumatism, malignancy and the like.

Accordingly, the present inventors have invented a novel low molecular compound that inhibits DDR2 tyrosine kinase activity, thereby confirming that they have an excellent therapeutic effect against diseases mediated by the activity of DDR2 such as cirrhosis, rheumatism or cancer. Completed.

The present invention, in order to treat diseases mediated by the tyrosine kinase activity of the DDR2 protein represented by cirrhosis, rheumatism, cancer and the like as described above, a novel small molecule compound having an inhibitory activity to DDR2 tyrosine kinase activity and DDR2 comprising the same It is an object to provide a therapeutic agent for a disease mediated by the tyrosine kinase activity of a protein.

The present invention relates to a novel furopyrimidine compound, a precursor thereof, a pharmaceutically acceptable salt thereof, and a tyrosine kinase activity of a DDR2 protein comprising the same, which inhibits tyrosine kinase activity of a DDR2 (Discoidin Domain Receptor 2) protein. It relates to the treatment of related diseases. Since the puropyridine compound of the present invention has the effect of inhibiting the activity of DDR2 tyrosine kinase, it can be used to treat diseases involving DDR2 tyrosine kinase activity, such as cirrhosis, rheumatoid arthritis, cancer and the like.

First, the present invention is a furopyrimidine compound defined by the following formula (1), a precursor thereof, and a pharmaceutically acceptable compound thereof, in which various derivatives are introduced into the furopyrimidine ring, which acts to inhibit the tyrosine kinase activity of the DDR2 protein. Provide salts:

[Formula 1]

In the above formula,

Z is O, S or NH,

n is an integer from 0 to 4,

R ₁ is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, amidine group, a haloalkyl group of C alkyl group of _{1 -C 4, C 1 -C 4} , C 1 - a C ₇ alkoxy group, C ₁ -C ₄ alkyl group, C ₁ alkylthio -C _4, C ₁ -C ₄ alkyl amide group, C ₁ -C ₄ acyl group, C ₁ -C ₄ Isiloxy group, C ₁ -C ₄ alkylsulfonamide group, C ₁ -C ₄ alkylsulfonate group, imidic acid C ₁ -C ₄ alkyl ester, thiimidic acid C ₁ -C ₄ alkyl ester, hydroxyna amino, Mo morpholine, acetate, acetamide, methanesulfonamide group is substituted with a methyl group, a halo-C ₁ -C ₄ alkyl group, an alkoxy group of C ₁ -C _4, or halogen-substituted one by groups substituted benzyloxy independently disubstituted Case,

The A ring represents benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, cyclohexyl, piperidine or morpholine,

R ₂ is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH ₂ , CSNH ₂ , alkyl group of C1-C5, haloalkyl group of C ₁ -C ₅ , alkyl ester, phenyl group, halogen substituted a phenyl group, a haloalkoxy group of C ₁ -C ₄ alkoxy group substituted with a phenyl group or a C ₁ -C ₄ group represents a substituted phenyl group,

R is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, amidine group, a haloalkyl group of C ₁ -C ₄ alkyl group, a C ₁ -C _4, C ₁ -C ₇ alkoxy, C ₁ -C ₄ alkylamino group import, C ₁ -C ₄ in the alkyl group, C ₁ -C ₄ alkyl amide group, an acylamino group of the C ₁ -C _4, a C ₁ -C ₄ acyl substituted by an alkyl sulfonamide of the oxy group or a C ₁ -C ₄ days or disubstituted independently represents a case.

In addition, the present invention is defined by the following formula (2) wherein the substituent R of the following formula (1), in which various derivatives are introduced into the furopyrimidine ring, which acts to inhibit the tyrosine kinase activity of the DDR2 protein Provided are furopyrimidine compounds, precursors thereof, and pharmaceutically acceptable salts thereof:

[Formula 4]

[Formula 2]

In the above formula,

Z is O, S or NH,

X is O, S or NH,

n is an integer from 0 to 4,

n 'is an integer from 0 to 4,

R ₁ and R ₃ are the same as or different from each other independently, and are an alkyl group of hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH ₂ , CSNH ₂ , amidine, C ₁ -C ₄ , alkyl amide of the C ₁ -C ₄ haloalkyl groups, C ₁ -C ₇ alkoxy group, C ₁ -C ₄ alkylamino group import, of C ₁ -C ₄ alkylthio, C ₁ -C ₄ group, C ₁ of the -C ₄ acylamino group, a C ₁ -C ₄ Isil oxy group, C ₁ -C ₄ alkyl sulfonamide group, a C ₁ -C ₄ alkyl sulfonate group, an already diksan C ₁ -C ₄ alkyl esters, thio already diksan C ₁ -C ₄ alkyl ester, hydroxy or amino, morpholine base, acetate, acetamide, methanesulfonamide group is substituted with a methyl group, an alkoxy group, a halogen of the haloalkyl group of C ₁ -C _4, C ₁ -C ₄ The case where it is mono-substituted or independently substituted by this substituted benzyloxy group is shown,

The A rings are each independently benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, cyclohexyl, piperidine or morpholine Ring represents pyrrole, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, piperidine or morpholine,

R ₂ is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, an alkyl group of _{C 1 -C 5, C 1 -C} 5 haloalkyl group of the alkyl ester, a phenyl group, a halogen This substituted phenyl group, the phenyl group substituted with the C ₁ -C ₄ alkoxy group, or the phenyl group substituted with the C ₁ -C ₄ haloalkoxy group is shown.

In addition, the present invention is a furropyrimidine compound defined by the following formula (3), a precursor thereof and a pharmaceutically acceptable, which introduce various derivatives to the furopyrimidine ring, which acts to inhibit the tyrosine kinase activity of DDR2 protein Provide salts:

[Formula 3]

In the above formula,

Z is O, S or NH,

n is an integer from 0 to 4,

R ₁ is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, amidine group, a haloalkyl group of C alkyl group of _{1 -C 4, C 1 -C 4} , C 1 - a C ₇ alkoxy group, C ₁ -C ₄ alkyl group, C ₁ alkylthio -C _4, C ₁ -C ₄ alkyl amide group, C ₁ -C ₄ acyl group, C ₁ -C ₄ Isiloxy group, C ₁ -C ₄ alkylsulfonamide group, C ₁ -C ₄ alkylsulfonate group, imidic acid C ₁ -C ₄ alkyl ester, thiimidic acid C ₁ -C ₄ alkyl ester, hydroxyna amino, Mo morpholine, acetate, acetamide, methanesulfonamide group is substituted with a methyl group, a halo-C ₁ -C ₄ alkyl group, an alkoxy group of C ₁ -C _4, or halogen-substituted one by groups substituted benzyloxy independently disubstituted Case,

The A ring represents benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, cyclohexyl, piperidine or morpholine,

R ₂ is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, an alkyl group of _{C 1 -C 5, C 1 -C} 5 haloalkyl group of the alkyl ester, a phenyl group, a halogen Represents a substituted phenyl group, a phenyl group substituted with a C ₁ -C ₄ alkoxy group or a phenyl group substituted with a C ₁ -C ₄ haloalkoxy group,

R is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, amidine group, a haloalkyl group of C ₁ -C ₄ alkyl group, a C ₁ -C _4, C ₁ -C ₄ alkoxy group, C ₁ -C ₄ alkyl group, C ₁ alkylthio -C _4, C ₁ -C ₄ alkyl amide group, an acylamino group of the C ₁ -C _4, C ₁ -C ₄ acyl substituted by an alkyl sulfonamide of the oxy group or a C ₁ -C ₄ days or disubstituted independently represents a case.

Compounds defined by Formula 1, Formula 2 or Formula 3 of the present invention may be in free form or in acid addition salt forms such as hydrochloric acid, sulfuric acid, citric acid or fumaric acid.

All compounds of the present invention can be synthesized from compounds represented by the following formula

[Formula IV]

In the case of the compound of formula (2), the substituent R 'of formula (IV) should be capable of producing a functional group which is substitutable by the substituent defined by formula (4), so that hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H , CONH ₂ , CSNH ₂ , amidine, C ₁ -C ₄ alkyl group, C ₁ -C ₄ haloalkyl group, C ₁ -C ₄ alkoxy group, C ₁ -C ₄ alkylamino group, C ₁ -C ₄ that the alkylthio group, an acylamino group, an acyloxy group or an alkyl sulfonamide group of the C ₁ -C ₄ a C ₁ -C ₄ a C ₁ -C ₄ alkyl amide group, C ₁ -C ₄ are preferred.

For example, the compound of formula 2 of the present invention may be prepared by using the compound of formula IV, wherein 'R' is -OCH ₃ (IV '), as Reaction Scheme 1 below:

Scheme 1

In the above formula,

Z is O, S or NH,

X is O, S or NH

n is an integer from 0 to 4,

R ₁ and R ₃ are each independently the same as or different from each other and are an alkyl group of hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH ₂ , CSNH ₂ , amidine, C ₁ -C ₄ , alkyl amide of the C ₁ -C ₄ haloalkyl groups, C ₁ -C ₄ alkoxy group, C ₁ -C ₄ alkylamino group import, of C ₁ -C ₄ alkylthio, C ₁ -C ₄ group, C ₁ of the -C ₄ acylamino group, a C ₁ -C ₄ Isil oxy group, C ₁ -C ₄ alkyl sulfonamide group, a C ₁ -C ₄ alkyl sulfonate group, an already diksan C ₁ -C ₄ alkyl esters, thio Imidic acid C ₁ -C ₄ alkyl ester, hydroxy or amino, morpholine, acetate, acetamide, methyl group substituted with methanesulfonamide group, haloalkyl group of C1-C4, alkoxy group of C1-C4, halogen substituted benzyl The case of mono- or independently substituted by an oxy group,

The A ring represents benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, cyclohexyl, piperidine or morpholine,

R ₂ is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, an alkyl group of _{C 1 -C 5, C 1 -C} 5 haloalkyl group of the alkyl ester, a phenyl group, a halogen This substituted phenyl group, the phenyl group substituted with the C ₁ -C ₄ alkoxy group, or the phenyl group substituted with the C ₁ -C ₄ haloalkoxy group is shown.

Scheme 1 is only one example of preparing Compound 2, and is prepared using Compound IV different from Scheme 1, or by adding isoamylnitrite and CCl ₄ to Compound VI ′ in formula -NH ₂ The reaction may proceed by dimethylating -OCH ₃ of compound VI 'without the process of preparing compound VII' substituted with -Cl.

In addition, the compound of Formula 1 or 3 can be prepared using compound IV of Formula IV having the same R 'as R as described above, since R' of the starting compound VI is maintained as it is. have.

For example, the compounds of formula 2 of the present invention may be prepared by the following scheme 2.

Scheme 2

As described above, Scheme 2 is just one example of preparing Compound 1, and isoamnitrite and CCl ₄ are not added to replace -NH ₂ of Compound IV '' with -Cl, and Compound IV ''. Compound 1 may also be prepared by reacting with Compound XII.

In addition, the compound of formula 3 of the present invention may be prepared, for example, by the following scheme 3:

Scheme 3

In Schemes 2 and 3,

Z is O, S or NH,

R ₁ is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, amidine group, a haloalkyl group of C alkyl group of _{1 -C 4, C 1 -C 4} , C 1 - a C ₇ alkoxy group, C ₁ -C ₄ alkyl group, C ₁ alkylthio -C _4, C ₁ -C ₄ alkyl amide group, C ₁ -C ₄ acyl group, C ₁ -C ₄ Isiloxy group, C ₁ -C ₄ alkylsulfonamide group, C ₁ -C ₄ alkylsulfonate group, imidic acid C ₁ -C ₄ alkyl ester, thiimidic acid C ₁ -C ₄ alkyl ester, hydroxyna amino, Mo morpholine, acetate, acetamide, methanesulfonamide group is substituted with a methyl group, a halo-C ₁ -C ₄ alkyl group, an alkoxy group of C ₁ -C _4, or halogen-substituted one by groups substituted benzyloxy independently disubstituted Case,

The A ring represents benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, cyclohexyl, piperidine or morpholine,

R ₂ is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, an alkyl group of _{C 1 -C 5, C 1 -C} 5 haloalkyl group of the alkyl ester, a phenyl group, a halogen Represents a substituted phenyl group, a phenyl group substituted with a C ₁ -C ₄ alkoxy group or a phenyl group substituted with a C ₁ -C ₄ haloalkoxy group,

R is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, amidine group, a haloalkyl group of C ₁ -C ₄ alkyl group, a C ₁ -C _4, C ₁ -C ₇ alkoxy, C ₁ -C ₄ alkylamino group import, C ₁ -C ₄ in the alkyl group, C ₁ -C ₄ alkyl amide group, an acylamino group of the C ₁ -C _4, a C ₁ -C ₄ acyl substituted by an alkyl sulfonamide of the oxy group or a C ₁ -C ₄ days or disubstituted independently represents a case.

In Scheme 2, n means an integer between 0 and 4.

In embodiments of the present invention, the compounds 1, 2 and 3 may be a compound of Nos. 5 to 115 in Table 6 below.

In addition, the present invention is a novel furopyrimidine derivative having the effect of inhibiting the activity of the DDR2 tyrosine kinase represented by the formula (1), (2), (3), (VI) and (VII) as an active ingredient and its pharmacologically acceptable Provided are DDR2 tyrosine kinase activity inhibitors comprising a therapeutically effective amount of one or more of the salts. In addition, the present invention comprises a therapeutically effective amount of at least one of the novel furopyrimidine derivatives represented by the formula (1), (2), (3), (VI) and (VII) as a active ingredient and pharmacologically acceptable salts thereof. It provides a therapeutic agent for diseases related to the activity of DDR2 tyrosine kinase, such as cirrhosis, rheumatism and the like. The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier, excipient, and the like. As can be seen from Table 7 below, not only the compounds of formulas 1 to 3, which are the final products of the present invention, but also the compounds of formulas VI and VII, which are intermediates, were found to have excellent DDR2 tyrosine kinase inhibitory activity. Therefore, the pharmaceutical composition of the present invention may include a compound of Formula VI and / or VII as an active ingredient.

As mentioned above, the DDR2 protein is a receptor protein having tyrosine kinase activity, and includes a tyrosine kinase active domain at the c-terminus. By using DNA recombination technology and baculovirus expression technology, it is possible to obtain a protein that is overexpressed in insect cells and has tyrosine kinase activity at the c-terminal site (amino acids 441-825), which is a cytoplasmic exposure site. It is also possible to enhance the kinase activity by inducing and modifying tyrosine phosphorylation in various ways to the kinase active site (Korean Patent Application No. 2002-0067233, “Modified DDR2 Tyrosine Kinase Active Protein” and Korean Patent Application No. 2003-0076967). See “Methods for modifying DDR2 tyrosine kinase activity and DDR2 proteins with kinase activity modified in this way”.

The inhibitory activity of the compounds of the present invention on DDR2 proteins having such tyrosine kinase activity was measured. Tyrosine kinase activity of the tyrosine kinase activity of the DDR2 protein can be measured in several ways. For example, biotin-attached poly (D4Y) n provided by Promega Corporation, histone H2B protein, or the degree of autophosphorylation of the tyrosine kinase active site of DDR2 protein may be measured as a peptide substrate. The transfer of phosphatase from ATP, which is a substrate of phosphate group, to a peptide, which is another substrate, can be measured by using a gamma position phosphate group of ATP after labeling with ³² P isotope, thereby detecting the activity of ³² P radioactive peptide substrate. Can be.

DDR2 tyrosine kinase inhibitory activity of the compounds synthesized in the present invention, by defining the concentration of each compound that inhibits DDR2 tyrosine kinase activity 50% by the IC ₅₀ value of each compound, it is shown in Table 7 below. As can be seen in Table 7, the compounds represented by the formulas (1), (2), and (3) as well as the formulas (VI) and (VII) all exhibit IC ₅₀ values of 500 uM or less, with excellent DDR2 protein tyrosine kinase inhibitory ability It can be seen that. Therefore, by the tyrosine kinase inhibitory activity of the DDR2 protein, the compound of the present invention can be used as a therapeutic agent for various diseases related to the tyrosine kinase activity of the DDR2 protein, for example, cirrhosis, rheumatism and cancer.

It has also been found that increased tyrosine kinase activity of DDR2 receptor proteins in hepatic stellate cells is associated with cell proliferation. Given that the dominant negative mutation of DDR2 receptor kinase inhibits the proliferation of hepatic stellate cells, inhibition of kinase activity of DDR2 protein is expected to inhibit the proliferation of hepatic stellate cells. Based on these facts, the results of various experiments have been conducted to verify the effects of inhibiting the proliferation and activity of hepatic stellate cells when the present compounds are treated in hepatic stellate cell culture.

Figure 1 shows that the compounds of the present invention inhibits tyrosine phosphorylation of DDR2 protein induced by collagen type 1 in HSC T6 cells, which is an activated hepatic astrocytic model (compound number 100 in Table 6). Was treated for 24 hours at a concentration of 5 uM to 20 uM, and the degree of tyrosine phosphorylation of DDR2 protein in HSC T6 cells was measured by Western blotting using phosphorylated tyrosine specific antibody. As shown in Figure 1, it can be seen that the tyrosine phosphorylation of DDR2 protein induced by collagen type 1, the active ligand of the DDR2 receptor, decreases depending on the treatment concentration.

Figure 2 shows that the compounds of the present invention selectively inhibit the growth of HSC T6 cells, the representative compounds of Table 6 of the present invention (No. 100) when treated 48 hours, HSC T6 ( It can be seen that the cell viability of-)-is significantly reduced compared to rat2 fibroblasts (-▼-) or HT1080 cells (...) with no expression of DDR2. As shown in Figure 2, it can be seen that the compound of the present invention selectively acts on cells expressing DDR2 through specific inhibition of DDR2 kinase activity.

The activation of hepatic stellate cells is considered to be a fundamental and main cause of cirrhosis of the liver. It is known that the number of hepatic stellate cells increases in liver cirrhosis and is activated at the same time. For this reason, removal of activated hepatic stellate cells from cirrhosis of the liver is thought to be a major strategy for achieving the therapeutic effect of liver cirrhosis. The compound of the present invention induces apoptosis of hepatic stellate cells and has an excellent therapeutic effect on liver cirrhosis. In order to confirm whether the compound of the present invention can actually induce apoptosis of hepatic stellate cells by inhibiting DDR2 kinase activity, the compound of the present invention (number 100 in Table 6) is treated to HSC T6 cells that are hepatic stellate cells. Was observed through observation of the DNA fragmentation phenomenon, the results are shown in FIG. As can be seen in Figure 3, the treatment of the compound of the present invention increased the DNA fragmentation phenomenon in proportion to the treatment concentration, which shows that the compound of the present invention can effectively remove the liver stellate cells.

As described above, the compounds of the present invention are expected to have an excellent therapeutic effect on cirrhosis by inhibiting the growth and activity of hepatic stellate cells, which are considered as the main cause of cirrhosis, and inducing apoptosis.

In addition, the compounds of the present invention can treat cirrhosis by inhibiting the accumulation of collagen in the liver, which causes liver fibrosis, which is known as another cause of cirrhosis. In order to confirm the effect of the compounds of the present invention, such as hepatic collagen accumulation inhibitory effect and liver cell damage caused by cirrhosis lesions, hepatic cured animal model by bile duct closure using a 7-week-old male whistar rat, Animals were administered a compound of the invention (number 100 in Table 6) once daily for three weeks at a dose of 50 mg / kg via intraperitoneal injection. Three weeks after the bile duct closure procedure, the compound of the present invention was administered to the group not to be administered (control), and the amount of hydroxyproline in the liver tissue, the amount of bilirubin in the blood, and the AST and ALT levels were measured. , The results are shown in Table 8 below in comparison with the values measured from the group not subjected to biliary duct repair.

First, the bilirubin level in the blood in the group that received the bile duct suture was higher than the normal group who did not receive the bile duct suture treatment, it was confirmed that the bile duct suture was successfully performed in the present invention. Hydroproline is an amino acid constituting collagen, which is hardly present in other proteins in the body, and in the present invention, the amount of hydroxyproline in liver tissue is presented as an indicator indicating directly the amount of collagen in liver tissue. Since AST or ALT enzymes usually increase their blood activity when liver damage occurs, their blood activity is proportional to the extent of liver damage, and thus is used as an index indicating the extent of liver damage.

As can be seen from Table 8, the amount of hydroxyproline in the group not administered the compound of the present invention after the bile duct suture procedure is increased by about 2.5 times compared to the group not receiving the bile duct suture procedure, which is bile duct suture Due to the progress of a significant amount of collagen accumulation in liver tissue to show that the progression of cirrhosis of the liver. In addition, the levels of bilirubin, AST and ALT in the blood were also significantly higher than those in the normal group who did not undergo bile duct closure, indicating that liver cirrhosis caused liver damage due to necrosis of liver cells.

On the other hand, as shown in Table 8, compared with the group left untreated for three weeks after the bile duct closure procedure, the group to which the compound of the present invention was administered increased the amount of hydroxyprorin in liver tissue or bilirubin, AST and ALT levels in blood. The results were markedly reduced. These results demonstrate that the compounds of the present invention inhibit the proliferation and activity inhibiting activity of hepatic stellate cells and inhibit the accumulation of collagen in liver tissues, thereby inhibiting the damage of liver tissues caused by cirrhosis lesions. Thus, it can be seen that the compounds of the present invention have excellent liver cirrhosis therapeutic effect.

In addition, the compounds of the present invention have excellent therapeutic effects on arthritis lesions such as rheumatism as well as cirrhosis. One feature of arthritis lesions such as rheumatism is the activation and hyperproliferation of synovial cells of the synovial membrane. Activated synovial cells intensify rheumatoid lesions by secreting excess protein to destroy collagen proteins that make up cartilage tissue.

Recently, increased expression of DDR2 protein in synovial cells during rheumatoid lesions was observed. In the sense that synovial cells are a kind of fibrotic cells as in the above-described hepatic stellate cells, it can be inferred that DDR2 expression is a etiological factor in synovial cells as in hepatic stellate cells.

In order to show that the compound of the present invention has a therapeutic effect against rheumatism by inhibiting the growth of synovial cells, the compound of the present invention (compound number 100 in Table 6) was applied to synovial fibroblasts extracted from rheumatoid patients for 48 hours by concentration. The viable cells after treatment and the number of cells immediately before compound treatment were compared by SRB quantification to obtain% cell viability and are shown in FIG. 4. As shown in FIG. 4, it can be seen that the cell viability of synovial fibroblasts decreases in proportion to the concentration of the compound of the present invention, which is a compound of the present invention which is a specific inhibitor of DDR2 kinase activity. It is shown that it has a therapeutic effect against rheumatism by inhibiting the growth and activity of synovial fibroblasts.

In addition, the compounds of the present invention have activity to inhibit the expression of matrix metalloproteinase-1 (MMP-1) in synovial fibroblasts, thereby having therapeutic activity against arthritis. FIG. 5 shows the results of quantifying MMP-1 m-RNA by conventional Northern blotting on synovial fibroblasts treated with the representative compounds of the present invention (Compound No. 100 in Table 6). It can be confirmed that MMP-1 expression is inhibited in synovial fibroblasts. This fact demonstrates that the compounds of the present invention can be used as a therapeutic agent for rheumatism lesions by inhibiting kinase activity of DDR2 and can be used as a therapeutic agent for malignancy through inhibition of cancer metastasis. As a further supporting result, the author of 'Identification of genes cell lines associated with the invasive status of human mammary carcinoma cell lines by transcriptional profiling' published in Tumor Biology 2003, pp. 189--198, by Vesna Evtimova et al. The results of the research paper can be cited. According to the study paper, it was confirmed that the amount of DDR2 gene expression in cancer cell lines with high metastasis activity increased with high correlation with each other.

Hereinafter, the compounds of the present invention, preparation methods and biological activities thereof will be described in more detail with reference to Examples, but the scope of the present invention is not limited to these Examples. In addition, the following examples illustrate some methods for preparing the compound, but the compounds of the present invention except for these are based on the method described in the following examples. It can be easily manufactured by itself.

EXAMPLE

I. Preparation of Compounds of the Invention

First, compounds I, II, III, and IV were prepared by the preparation process as in Scheme 4:

Scheme 4

Example 1: Preparation of 4-methoxyphenylacetylchloride (Compound I)

After dissolving 10 ml of benzene in 16.62 g (0.1 mol) of 4-methoxyphenylacetic acid, 29 ml (0.2 mol) of thionyl chloride (SOCl ₂ ) was added, and the mixture was heated to reflux and stirred for 1 hour. Thionyl chloride and the solvent remaining in the reaction solution were concentrated to remove the product 4-methoxyphenylacetylchloride (Compound I) in a quantitative yield to give a liquid that was used without purification.

Example 2: Preparation of 1-phenyl-2- (4-methoxyphenyl) ethanone (Compound II)

To 40 g (0.3 mol) of aluminum (III) chloride, 200 ml of dichloromethane was added and stirred. Here, a dilution of 29.5 ml (0.33 mol) of benzene and 18.5 g (0.1 mol) of 4-methoxyphenylacetyl chloride obtained in Example 1 in 100 ml of dichloromethane (CH ₂ Cl ₂ ) was performed for about 30 minutes. Slowly added and stirred at room temperature for 2 hours. The obtained reaction solution was added to 400 ml of 1 M aqueous hydrochloric acid solution, the organic layer was extracted, dried and concentrated, and then purified by column chromatography to give 1-phenyl-2- (4-methoxyphenyl) ethanone (compound) as an off-white solid product. II). In the same manner, compound 1- (4-tolyl) -2- (4-methoxyphenyl) ethanone and 1- (4-tolyl) -2- (4-cyanophenyl) ethanone were obtained.

Example 3: Preparation of Alpha-Bromoketone Compound (Compound III)

(1) 2-bromo-4'-chloropropiophenone (Compound IIId)

50 ml of acetic acid was added dropwise to 16.86 g (0.1 mol) of 4'-chloropropiophenone, cooled to 0 ° C, and 5.14 ml (0.1 mol) of bromine was slowly added dropwise. After stirring at room temperature for 2 hours, 270 ml of water were added, filtered and washed with sufficient water and dried to give 24.13 g of the product 2-bromo-4'-chloropropiophenone (IIId) as a white solid. In the same manner, compounds IIIf to IIIi, compound IIIk, compound IIIl, compound IIIn, and compound IIIu of Table 1 were obtained, respectively.

(2) 2-bromo-4'-methoxypropiophenone (Compound IIIe)

To 32.84 g (0.2 mol) of 4'-methoxypropiophenone and 89.34 g (0.4 mol) of CuBr ₂ were added dropwise 450 ml of dioxane, followed by heating to reflux for 6 hours. After cooling to room temperature to remove dioxane, water was added and the organic layer obtained by extraction 2-3 times with ethyl acetate was dried and concentrated, and recrystallized with n-hexane to give the product 2-bromo-4'- as an off-white solid. 48.62 g of methoxypropiophenone (IIIe) were obtained.

(3) 2-bromo-1- (4'-chlorophenyl) -3-methyl-1-butanone (compound IIIm)

14 ml of carbon disulfide was added dropwise to 1.02 ml (10 mmol) of chlorobenzene, and 1.6 g (12 mmol) of aluminum (III) chloride was added thereto, followed by heating to reflux. A solution of 1.46 ml (10 mmol) of isovaleryl chloride in 2 ml of carbon disulfide was slowly added thereto for about 30 minutes, heated for 6 hours, and then cooled to room temperature and concentrated. An aqueous 1 M hydrochloric acid solution was added to the obtained reaction solution, and the organic layer obtained by extracting 2-3 times with diethyl ether was washed with 10% aqueous sodium hydroxide solution, dried and concentrated, and purified by column chromatography to give the product 1 as a white solid. 1.85 g of-(4'-chlorophenyl) -3-methyl-1-butanone were obtained.

¹ H NMR (400 MHz DMSO- d ₆ ) δ 7.89 (d, J = 8.4 Hz, 2H), 7.43 (d, J = 8.4 Hz, 2H), 2.81 (d, J = 6.8 Hz, 2H), 2.36 ~ 2.20 (m, 1H), 0.99 (d, J = 6.4 Hz, 6H)

To 1.85 g (9.4 mmol) of 1- (4'-chlorophenyl) -3-methyl-1-butanone and 2.1 g (9.4 mmol) of CuBr _2, 14 ml of ethyl acetate and 14 ml of chloroform were added dropwise. Heated to reflux for 2 hours. After cooling to room temperature to remove the solid, the filtrate was dried and concentrated to give the product 2-bromo-1- (4'-chlorophenyl) -3-methyl-1-butanone (IIIm) as a white solid.

(4) 2-bromo-4'-phenoxypropiophenone (Compound IIIj)

100 ml of dimethylaniline was added dropwise to 13.2 ml (150 mmol) of phenol, 16.6 g (120 mmol) of potassium carbonate (K ₂ CO ₃ ) and 19.4 ml (140 mmol) of 4'-fluoropropiophenone, and heated for 4 hours. It was refluxed. After cooling to room temperature and adding water, the organic layer obtained by extraction 2-3 times with diethyl ether was dried and concentrated, purified by column chromatography to obtain 27.6 g of a product 4'-phenoxypropiophenone as a white solid. .

¹ H NMR (400 MHz DMSO) δ 7.99 (d, J = 8.8 Hz, 2H), 7.49-7.43 (m, 2H), 7.28-7.22 (m, 1H), 7.15-7.10 (m, 2H), 7.04 ( d, J = 8.8 Hz, 2H), 3.00 (q, J = 6.8 Hz, 2H), 1.07 (t, J = 6.8 Hz, 3H)

11.3 g (50 mmol) of 4'-phenoxypropiophenone obtained as described above and 50 mg (0.4 mmol) of aluminum (III) chloride were added to 20 ml of diethyl ether, and the bromine was slowly cooled after cooling to 0 ° C. Added dropwise. After stirring for 3 hours at the same temperature, water was added and the organic layer obtained by extraction with diethyl ether was dried and concentrated. Recrystallization with n-hexane afforded 2-bromo-4'-phenoxypropiophenone (IIIj).

The analysis results of the alpha-bromoketone compounds IIId to IIIu obtained in Example 3 are shown in Table 1 below.

R R ₂ ¹ H NMR (400 MHz DMSO-d ₆ ) ?? (ppm) yield(%) IIId Cl CH ₃ 7.97 (d, J = 9.2 Hz, 2H), 7.46 (d, J = 9.2 Hz, 2H), 5.23 (q, J = 6.8 Hz, 1H), 1.90 (d, J = 6.8 Hz, 3H) 97 IIIe OCH ₃ CH ₃ 8.03 (d, J = 8.8 Hz, 2H), 7.08 (d, J = 8.8 Hz, 2H), 5.79 (q, J = 6.4 Hz, 1H), 3.87 (s, 3H), 1.77 (d, J = 6.4 Hz, 3H) 98 IIIf CH ₃ CH ₃ 7.94 (d, J = 8.4 Hz, 2H), 7.35 (d, J = 8.4 Hz, 2H), 5.79 (q, J = 6.8 Hz, 1H), 2.38 (s, 3H), 1.76 (d, J = 6.8 Hz, 3H) 79 IIIg F CH ₃ ¹ H NMR (400 MHz CDCl ₃ ) ?? 8.08 ~ 8.05 (m, 2H), 7.18 ~ 7.14 (m, 2H), 5.24 (q, J = 6.4 Hz, 1H), 1.91 (d, J = 6.4 Hz, 3H) 67 IIIh I CH ₃ ¹ H NMR (400 MHz CDCl ₃ ) ?? 7.86 (d, J = 8.4 Hz, 2H), 7.73 (d, J = 8.4 Hz, 2H), 5.21 (d, J = 6.4 Hz, 1H), 1.89 (d, J = 6.4 Hz, 3H) 40 IIIi Br CH ₃ 7.98 (d, J = 8.4 Hz, 2H), 7.79 (d, J = 8.4 Hz, 2H), 5.82 (q, J = 6.4 Hz, 1H), 1.79 (d, J = 6.4 Hz, 3H) 98 IIIj OPh CH ₃ 8.07 (d, J = 8.4 Hz, 2H), 7.58-7.40 (m, 2H), 7.32-6.98 (m, 5H), 5.78 (q, J = 6.4 Hz, 1H), 1.77 (d, J = 6.4 Hz , 3H) 76 IIIk SCH ₃ CH ₃ 7.94 (d, J = 8.4 Hz, 2H), 7.28 (d, J = 8.4 Hz, 2H), 5.25 (q, J = 6.8 Hz, 1H), 2.53 (s, 3H), 1.89 (d, J = 6.8 Hz, 3H) 57 IIIl Cl Et ¹ H NMR (400 MHz CDCl ₃ ) ?? 7.96 (d, J = 8.8 Hz, 2H), 7.46 (d, J = 8.8 Hz, 2H), 5.50 (t, J = 6.4 Hz, 1H), 2.23-2.13 (m, 2H), 1.09 (t, J = 7.6, 3H) 40 IIIm Cl CH (CH ₃ ) ₂ 8.08 (d, J = 8.4 Hz, 2H), 7.65 (d, J = 8.4 Hz, 2H), 5.62 (d, J = 8 Hz, 1H), 2.20-2.06 (m, 1H), 1.11 (d, J = 6.4 Hz, 3H), 0.99 (d, J = 6.4 Hz, 3H) 64 IIIn Cl Ph 8.10 (d, J = 8.4 Hz, 2H), 7.72 ~ 7.24 (m, 7H), 7.20 (s, 1H) 95 IIIu OCH ₃ ¹ H NMR (400 MHz CDCl ₃ ) ?? 7.97 (d, J = 8.0 Hz, 2H), 7.41 (d, J = 8.8 Hz, 2H), 7.16 (d, J = 8.0 Hz, 2H), 6.89 (d, J = 8.8 Hz, 2H), 6.36 ( s, 1H), 3.85 (s, 3H), 2.33 (s, 3H) 99

In addition, starting material IV was prepared by the following scheme 5:

Scheme 5

Example 4 Preparation of Alpha-Hydroxyketone (Compound V) Compound

(1) Preparation of 4,4'-dichlorobenzoin (Vv)

120 ml of ethyl alcohol was added dropwise to 14 g (0.1 mol) of chlorobenzaldehyde, and then a solution of 14.98 g (0.23 mol) of potassium cyanide (KCN) dissolved in 10 ml of water was slowly added thereto, followed by heating to reflux for 12 hours. After cooling to room temperature, water was added, followed by filtration and drying and purification by recrystallization or column chromatography to give the product 4,4'-dichlorobenzoin (Vv) as a pale yellow solid.

(2) Preparation of 2-hydroxy-2-phenyl- (4-methoxy) acetophenone (Vx)

50 ml of ethyl alcohol was added to 2.12 g (10 mmol) of benzoin, and 1.36 g (2 mmol) of anisealdehyde and 30 ml of saturated potassium cyanide (KCN) solution were added thereto, and the mixture was heated to reflux and stirred for 3 hours. Water was added to the reaction solution, and the organic layer obtained by extraction with ethyl acetate two to three times was dried and concentrated, and then purified by column chromatography to give the product as a pale, yellow solid 2-hydroxy-2-phenyl- (4-meth). Methoxy) acetophenone (Vx) was obtained. In the same manner, the compounds Vq and Vr in Table 2 were obtained, respectively.

The analysis results of the alpha-hydroxy ketone compounds Vv to Vr obtained in Example 4 are shown in Table 2 below.

Example 5: Preparation of 2-amino-3-furonitrile compound (Compound IV)

(1) Preparation of 2-amino-3-cyano-5- (4-chlorophenyl) furan (IVa)

In 11.67 g (50 mmol) of 2-bromo-4'-chloroacetophenone (IIIa) and 3.3 g (50 mmol) of malononitrile (CH ₂ (CN) ₂ ) obtained in Example 3, dimethylformamide ( DMF) 20 ml was added dropwise, cooled to 0 ° C., and 15.5 ml (150 mmol) of diethylamine (Et ₂ NH) were added slowly for about 30 minutes. At this time, the temperature of the reaction solution was set not to exceed 40 degreeC. Then, after stirring at room temperature for 3 hours, 60 ml of water was added thereto, filtered, washed well with water and n-hexane, and dried. Recrystallization with ethyl alcohol gave the product 2-amino-3-cyano-5- (4-chlorophenyl) furan (IVa) as a tan solid. Compounds IVb to IVp and Compounds IVs to IVu in Table 3 below were obtained in the same manner.

(2) Preparation of 2-amino-3-cyano-4,5-di (4-chlorophenyl) furan (IVv)

To 7 g (25 mmole) of 4,4'-dichlorobenzoin (Vv) and 1.98 g (30 mmol) of malononitrile obtained in Example 4, 15 ml of dimethylformamide (DMF) was added, followed by cooling to 0 ° C. 100 mmol of diethylamine was slowly added dropwise for about 30 minutes. At this time, the temperature of the reaction liquid did not exceed 40 degreeC. After stirring for 18 hours at room temperature, 100 ml of water was added thereto, filtered, washed well with water and n-hexane, and dried. Then, it recrystallized with diethyl ether, and the product 2-amino-3-cyano-4,5-di (4-chlorophenyl) furan (IVv) of a tan solid was obtained. In the same manner, compounds IVq, IVr, and IVw of Table 3 were obtained, respectively.

The analysis results of the 2-amino-3-furonitrile-based compounds IVa to IVw obtained above are shown in Table 3 below.

Using Compound IV obtained in Example 5 as a starting material, 4-amino furypyrimidine-based compound VI and 4-chloro-pyropyrimidine-based compound VII were prepared as in Scheme 6 below:

Scheme 6

Example 6 Preparation of 4-Amino Furopyrimidine Compound 4-Amino-6- (4-chlorophenyl) furo [2,3, d] pyrimidine (VIa)

To 4.37 g (20 mmol) of 2-amino-3-cyano-5- (4-chlorophenyl) furan (IVa) obtained in Example 5, 20 ml of formamide was added dropwise and heated to reflux for 12 hours. Cooled to. 100 ml of water was added thereto, and the resulting solid was filtered, washed well with water and n-hexane, and dried. Then recrystallized with acetone and ethyl alcohol to give the product 4-amino-6- (4-chlorophenyl) furo [2,3, d] pyrimidine (VIa) as a brown solid. In the same manner, the following compounds VIb to VIw of Table 4 were obtained, respectively.

The analysis results of 4-amino furypyrimidine-based compounds VIa to VIw obtained as described above are shown in Table 4 above:

Example 7 Preparation of 4-Chloropyropyrimidine Compound 4-Chloro-6- (4-chlorophenyl) furo [2,3, d] pyrimidine (VIIa)

50 ml of chloroform was added dropwise to 7.37 g (30 mmol) of 4-amino-6- (4-chlorophenyl) -furo [2,3, d] pyrimidine (VIa) obtained in Example 6, and isoamylnitrite 8.6 ml (65.1 mmol) were added and heated to reflux for 14 hours. Then, cooled to room temperature, chloroform was removed under reduced pressure and purified by column chromatography to give the product 4-chloro-6- (4-chlorophenyl) furo [2,3, d] pyrimidine (VIIa) as a yellow solid. Got it. In the same manner, the compounds VIIb to VIIw of Table 5 below were obtained, respectively.

The analysis results of the 4-chlorofuropyrimidine-based compounds VIIa to VIIw obtained as described above are shown in Table 5 below.

Example 8

Using Compound VII obtained as described above, Compound A, which is a precursor of the compound of the present invention, can be prepared, for example, by Scheme 7 below.

Scheme 7

Example 8-1 Preparation of 4-chloro-5-methyl-6- (4-hydroxyphenyl) furo [2,3, d] pyrimidine (VIII)

10 ml of dichloromethane was added to 1.268 g (4.6 mmol) of 4-chloro-5-methyl-6- (4-methoxyphenyl) furo [2,3, d] pyrimidine (VIIe) obtained in Example 7. After cooling to -78 ° C, 13.85 ml (1M in dichloromethane) BBr ₃ was slowly added dropwise. Then, after stirring for 12 hours at room temperature, saturated sodium bicarbonate (Na ₂ HCO ₃ ) Aqueous solution was slowly added, the organic layer obtained by extraction 2-3 times with ethyl acetate was washed with brine, dried and concentrated and purified by column chromatography to give the yellow solid product 4-chloro-5-methyl-6- ( 989 mg of 4-hydroxyphenyl) furo [2,3, d] pyrimidine was obtained. (Yield 82%)

¹ H NMR (400 MHz DMSO- d ₆ ) δ 10.13 (s, 1H), 8.75 (s, 1H), 7.67 (d, J = 8.8 Hz, 2H), 6.97 (d, J = 8.8 Hz, 2H), 2.56 (s, 3 H)

Example 8-2 Preparation of 4-chloro-5-methyl-6- [4- (2-chloroethoxy) phenyl] furo [2,3, d] pyrimidine (IX ')

859 mg (3.3 mmol) of 4-chloro-5-methyl-6- (4-hydroxyphenyl) furo [2,3, d] pyrimidine (7 ') obtained in Example 8-2, cesium carbonate (Cs 2.32 g (7.26 mmol) of ₂ CO ₃ ) and 1.32 ml (7.26 mmol) of 2-chloroethyl para-toluenesulfonate were added to a mixture of 5 ml of dimethylformamide, followed by stirring at room temperature for 2 hours. 20 ml of water was added to the reaction solution, which was filtered and washed with ethyl acetate. The organic layer obtained by extracting the filtrate two to three times with ethyl acetate was dried and concentrated, and then purified by column chromatography to give a white solid product 4-chloro-5-methyl-6- [4- (2-chloroethoxy) 205 mg of phenyl] furo [2,3, d] pyrimidine (corresponding to compound XI ′ of Scheme 7 above when n = 1 in compound XI of Scheme 1) was obtained. (Yield: 19%)

¹ H NMR (400 MHz CDCl ₃ ) δ 8.69 (s, 1H), 7.76 (d, J = 8.8 Hz, 2H), 7.07 (d, J = 8.8 Hz, 2H), 4.32 (t, J = 6.0 Hz, 2H), 3.89 (t, J = 6.0 Hz, 2H), 2.64 (s, 3H)

Example 8-3: of 4- (3-hydroxyanilino) -5-methyl-6- [4- (2-chloroethoxy) phenyl] furo [2,3, d] pyrimidine (X ') Produce

79 mg (0.25 mmol) of 4-chloro-5-methyl-6- [4- (2-chloroethoxy) phenyl] furo [2,3, d] pyrimidine (8 ') obtained in Example 8-2. 2 ml of n-butyl alcohol was added to 53.5 mg (0.5 mmol) of 3-aminophenol and heated to reflux for 4 hours. After the solvent was removed, water was added, and the organic layer obtained by extraction with ethyl acetate two to three times was dried and concentrated. Then, the residue was purified by column chromatography to give the brown solid product 4- (3-hydroxyanilino) -5-methyl-6- [4- (2-chloroethoxy) phenyl] furo [2,3, d ] 80 mg of pyrimidine (compound X 'of Scheme 7: n = 1 in compound X of Scheme 1) was obtained. (Yield 83%)

¹ H NMR (400 MHz DMSO- d ₆ ) δ 9.42 (s, 1H), 8.51 (s, 1H), 8.36 (s, 1H), 7.69 (d, J = 8.8 Hz, 2H), 7.26 to 7.04 (m , 5H), 6.52 (d, J = 8.0 Hz, 1H), 4.35 (br t, 2H), 4.00 (br t, 2H), 2.60 (s, 3H)

Example 8-4: 4- (3-hydroxyanilino) -5-methyl-6- {4- [2- (4-morpholinyl) ethoxy] phenyl} furo [2,3, d] pyri Preparation of Midine (A)

4- (3-hydroxyanilino) -5-methyl-6- [4- (2-chloroethoxy) phenyl] furo [2,3, d] pyrimidine (9 ') obtained in Example 8-3. Excess morpholine 0.07 ml (0.8 mmol) was added to 19 mg (0.048 mmol) and 7 mg (0.046 mmol) of sodium iodide (NaI), followed by stirring at 90 ° C. for 24 hours. Saturated sodium hydrogen carbonate (Na ₂ HCO ₃ ) to the reaction An aqueous solution was added, the organic layer obtained by extraction 2-3 times with ethyl acetate was dried and concentrated, and then purified by column chromatography to give a yellow solid product 4- (3-hydroxyanilino) -5-methyl-6- 15 mg of {4- [2- (4-morpholinyl) ethoxy] phenyl} furo [2,3, d] pyrimidine (A) were obtained. (Yield: 71%)

¹ H NMR (400 MHz DMSO- d ₆ ) δ 9.42 (s, 1H), 8.50 (s, 1H), 8.36 (s, 1H), 7.67 (d, J = 8.8 Hz, 2H), 7.24-7.00 (m , 5H), 6.51 (d, J = 8.0 Hz, 1H), 4.17 (t, J = 5.6 Hz, 2H), 3.59 (t, J = 4.0 Hz, 2H), 2.73 (t, 2H), 2.59 (s , 3H), 2.50 (br t, 4H)

Example 9 Preparation of 4,5,6-Substituted Furopyrimidine Compounds

Compound Puropyrimidine-based compound of the present invention was prepared using compound VI or VII obtained in Example 6 or 7 as a starting material.

Scheme 8

(1) Preparation of 4-anilino-6- (4-chlorophenyl) furo [2,3, d] pyrimidine (5)

79.5 mg (0.3 mmol) of 4-chloro-6- (4-chlorophenyl) furo [2,3, d] pyrimidine (VIIa) and 55.9 mg (0.6 mmol) of aniline are n-butyl alcohol (or ethyl alcohol or iso Propyl alcohol) 2 ml was added dropwise, heated to reflux for 4 hours, and then the solvent was removed. Dissolved in 1 ml of dimethyl sulfoxide, added to 15 ml of water, filtered, washed thoroughly with water and n-hexane and dried to give an off-white solid product 4-anilino-6- (4-chlorophenyl) furo [2,3, d ] Pyrimidine (5) was obtained. In the same manner, the compounds 6 to 64, 85, 89, 91 to 93, and 107 to 111 in Table 6 below were obtained, respectively.

(2) Preparation of 4- (4-methoxybenzoylamino) -6- (4-chlorophenyl) furo [2,3, d] pyrimidine (66)

To 245.66 mg (1 mmol) of 4-amino-6- (4-chlorophenyl) furo [2,3, d] pyrimidine (VIa) prepared in Example 6, 1 ml of pyridine and 511.8 mg of 4-methoxybenzoyl chloride (3 mmol) was added thereto, heated to reflux for 2 hours, and then pyridine was removed under reduced pressure. 1N aqueous sodium hydroxide solution was added, the organic layer obtained by extraction with ethyl acetate two to three times was washed with brine, dried and concentrated, and then purified by column chromatography to give a brown solid product 4- (4-methoxybenzoylamino)- 6- (4-chlorophenyl) furo [2,3, d] pyrimidine (66) was obtained. In the same manner, the compounds 65 in Table 6 and 67 to 73 were obtained, respectively.

(3) Preparation of 4- (3-methoxyphenoxy) -5-methyl-6- (4-chlorophenyl) furo [2,3, d] pyrimidine (74)

2 ml of tetrahydrofuran (THF) was added dropwise to 37.2 mg (0.3 mmol) of 3-methoxyphenol and 24 mg (0.6 mmol) of sodium hydride (60% NaH) were added, followed by stirring at room temperature for about 10 minutes. . 0.3 mmol of 4-chloro-5-methyl-6- (4-chlorophenyl) furo [2,3, d] pyrimidine (VIId) prepared in Example 7 was added to the reaction solution, which was then stirred at room temperature for 2 hours. After cooling, 10 ml of water was slowly added dropwise. The resulting solid was filtered, washed and dried with sufficient water and n-hexane to give the brown solid product 4- (3-methoxyphenoxy) -5-methyl-6- (4-chlorophenyl) furo [2,3, d] pyrimidine (74) was obtained. In the same manner, the compounds 75 to 78 of Table 6 were obtained, respectively.

(4) Preparation of 4- (3-hydroxyanilino) -5,6-di (4-hydroxyphenyl) furo [2,3, d] pyrimidine (90)

150 mg (0.31 mmol) of 4- (3-hydroxyanilino) -5,6-di (4-methoxyphenyl) furo [2,3, d] pyrimidine (89) prepared above and pyridine chloride 752 A mixture of mg (6.51 mmol) was heated to reflux at 210 ° C. for 3 hours. Water was added to the reaction product, the organic layer obtained by extraction with ethyl acetate two to three times was dried and concentrated, and then purified by column chromatography to give a brown solid product 4- (3-hydroxyanilino) -5,6- 15 mg of di (4-hydroxyphenyl) furo [2,3, d] pyrimidine (90) were obtained.

(5) Preparation of 4- (2-pyridylamino) -5-methyl-6- (4-chlorophenyl) furo [2,3, d] pyrimidine (96)

265.1 mg (1 mmol) of 4-chloro-5-methyl-6- (4-chlorophenyl) furo [2,3, d] pyrimidine (VIId) prepared in Example 7 and 141.17 mg of 2-aminopyridine ( 1.5 mmol) was dissolved in 3 ml of dimethylformamide. The mixture was cooled to 0 ° C., and 80 mg (2 mmol) of sodium hydride (60% NaH) was added thereto, followed by stirring at room temperature for 3 to 8 hours. Aqueous solution of saturated ammonium chloride (NH ₄ Cl) was added to the reaction solution, and the organic layer obtained by extraction with ethyl acetate two to three times was dried and concentrated, and then purified by column chromatography to give a brown solid product 4- (2-pyri). Dylamino) -5-methyl-6- (4-chlorophenyl) furo [2,3, d] pyrimidine (96) was obtained. In the same manner, the compounds 94, 95, and 97 to 106 of Table 6 were obtained, respectively.

(6) Preparation of 4- ( trans -4-hydroxycyclohexylamino) -5-methyl-6- (4-chlorophenyl) furo [2,3, d] pyrimidine (113)

265.1 mg (1 mmol) of 4-chloro-5-methyl-6- (4-chlorophenyl) furo [2,3, d] pyrimidine (VIId) prepared in Example 7 and trans -4-aminocyclohexane To a mixture of 303.3 mg (2 mmol) of all chloride in 3 ml of n-butyl alcohol was added 2.2 mmol of triethylamine (Et ₃ NH), followed by heating to reflux and stirring for 3 to 12 hours. Aqueous solution of saturated ammonium chloride (NH ₄ Cl) was added to the reaction solution, and the organic layer obtained by extracting with ethyl acetate two to three times was dried and concentrated, and then purified by column chromatography to obtain 4- ( trans -4-hydroxy). Cyclohexylamino) -5-methyl-6- (4-chlorophenyl) furo [2,3, d] pyrimidine (113) was obtained. In the same manner, the compounds 112, 114, and 115 in Table 6 were obtained, respectively.

(7) Preparation of 4- (3-hydroxyphenoxy) -5-methyl-6- (4-chlorophenyl) furo [2,3, d] pyrimidine (79)

In 366.86 mg (1 mmol) of 4- (3-methoxyphenoxy) -5-methyl-6- (4-chlorophenyl) furo [2,3, d] pyrimidine (74) prepared above, dichloromethane ( 5 ml of CH ₂ Cl ₂ ) were added, cooled to −78 ° C., and 0.25 ml of BBr ₃ (1M in dichloromethane) was slowly added dropwise. After stirring for 12 hours at room temperature, saturated sodium bicarbonate (Na ₂ HCO ₃ ) The resulting organic layer was slowly added to an aqueous solution, extracted two or three times with ethyl acetate, washed with brine, dried and concentrated, purified by column chromatography, and then purified by column chromatography (4- (3-hydroxyphenoxy) -5-methyl-6). -(4-chlorophenyl) furo [2,3, d] pyrimidine (79) was obtained. Compounds 80-84, 86, and 87 were obtained in the same manner using compounds 19, 69, 27, 25, 22, 59, and 62 in Table 6 below.

The analysis results of the 4,5,6-substituted furopyrimidine-based compound obtained above are shown in Tables 6-a to 6-j below.

II. Assessment of Biological Activity of Compounds of the Invention

Hereinafter, the experiments to evaluate the biological activity of DDR2 for the compound obtained above will be described in detail.

Example 10 Determination of Inhibitory Activity on the Tyrosine Kinase Activity of the DDR2 Protein of the Compound of the Present Invention

Tris-HCl (pH 7.5), MgCl 2 5 mM, 100 ng of activated DDR2 kinase enzyme protein (prepared by the method described in Korean Patent Application No. 2002-0067233 and Korean Patent Application No. 2003-0076967), ATP 10 uM and In a mixed solution of 0.2 uCi of P32-gamma-ATP 0.2 uCi, reaction was performed for 10 to 30 minutes using 2 ug of poly (D4Y) n (Promega, USA) or histone H2B protein with biotin as a peptide substrate, and then 1/2 The reaction was terminated by addition of a volume of 30% phosphoric acid solution.

The reaction solution was spotted on avidin-coated membranes (Promega, USA) when biotin-attached poly (D4Y) n was used as the peptide substrate, and on p81 cellulose paper when histone was used as the substrate. It was. After washing 5 times with 10 mM Tris-HCl (pH 8.0) and 100 mM NaCl solution, the activity generated from the phosphorylated peptide attached to the membrane was measured by BAS radiographic imaging (Kodak) to quantify the degree of enzyme reaction. The tyrosine kinase activity of DDR2 protein was measured.

To measure the autophosphorylation activity of the DDR2 kinase active site, the reaction was carried out in the reaction solution except for the peptide substrate under the above conditions, and the reaction was electrophoresed on a 10% PAGE gel, followed by staining with Kumaji to confirm the presence of the DDR2 kinase active site protein. Thereafter, the gel was dried and subjected to autoradiography using an X-ray film to measure the degree of autophosphorylation of the DDR2 protein site.

In order to measure the inhibitory activity of the compound, the compound dissolved in DMSO was previously added to the enzyme reaction solution at various concentrations, and then the enzyme reaction was carried out by adding the DDR2 kinase enzyme to measure the degree of inhibition of the enzyme activity at the specific concentration of each compound. In addition, the concentration of each compound that inhibits 50% enzymatic activity was determined as IC ₅₀ (concentration value that inhibited 50% activity) of the compound, and is shown in Table 7 below.

compound IC ₅₀ (uM) compound IC ₅₀ (uM) compound IC ₅₀ (uM) compound IC ₅₀ (uM) compound IC ₅₀ (uM) compound IC ₅₀ (uM) VIa <100 VIIe <500 16 <100 42 <100 68 <500 94 <100 VIb <100 VIIf <500 17 <100 43 <100 69 <500 95 <500 VIc <100 VIIg <500 18 <500 44 <100 70 <500 96 <100 VId <100 VIIh <500 19 <500 45 <100 71 <500 97 <100 VIe <100 VIIi <500 20 <100 46 <500 72 <500 98 <100 Vif <100 VIIj <500 21 <500 47 <100 73 <100 99 <500 VIg <100 VIIk <500 22 <100 48 <100 74 <100 100 <500 VIh <100 VIIl <500 23 <100 49 <100 75 <500 101 <500 VIi <100 VIIm <500 24 <500 50 <100 76 <500 102 <500 VIj <100 VIIo <500 25 <500 51 <500 77 <500 103 <500 VIk <100 VIIr <500 26 <500 52 <100 78 <500 104 <100 VIl <100 VIIs <500 27 <500 53 <100 79 <100 105 <100 VIm <100 VIIu <500 28 <100 54 <100 80 <500 106 <100 VIn <100 VIIv <500 29 <100 55 <100 81 <500 107 <500 VIx <100 VIIw <500 30 <100 56 <500 82 <100 108 <500 VIq <100 5 <500 31 <500 57 <100 83 <100 109 <500 VIr <100 6 <500 32 <500 58 <100 84 <100 110 <500 VIs <100 7 <500 33 <500 59 <100 85 <100 111 <100 VIt <100 8 <100 34 <500 60 <100 86 <100 112 <500

As can be seen in Table 7, the compound of the present invention was shown to effectively inhibit the tyrosine kinase activity of DDR2 protein. Thus, the compounds of the present invention can be expected to have a therapeutic effect against liver metastasis, rheumatism and metastasis caused by the tyrosine kinase activity of DDR2 protein.

In the following experiments, although the number 100 compound of Table 6 was used as the representative compound of the compound of the present invention, as shown in Table 7, the effect on the representative compound is also applied to other compounds of the present invention. It can be predicted.

Example 11 Measurement of Tyrosine Phosphorylation Inhibitory Activity of DDR2 Protein in HSC T6 Cells of Compounds of the Invention

Compounds of the invention treated HSC T6 cells (Professor Friedman, Mount Sinai Medical School, New York, USA) 24 hours at a collagen 10 ug / ml concentration or dissolved in DMSO with a 10 ug / ml collagen concentration (number in Table 6) 100) were treated for 24 hours at different concentrations (0, 5, 10 and 20 uM), and then cells were recovered and disrupted with 1x lameli solution and electrophoresed on a 7% PAGE gel. . After transferring the developed protein of the gel to the nitro cellulose paper, Western brotting was performed using an antibody that specifically recognizes human DDR2 and an antibody that specifically recognizes phosphorylated tyrosine, and the results are shown in FIG. 1. It was.

As can be seen in Figure 1, the compound of the present invention was shown to decrease the amount of phosphorylated DDR2 protein induced by collagen in the presence of tyrosine, and the degree of decrease increases with increasing treatment concentration. Therefore, it can be confirmed that the compound of the present invention has inhibitory activity against tyrosine phosphorylation of DDR2 protein in HSC T6 cells.

Example 12 Measurement of Cell Proliferation Inhibitory Activity of Compounds of the Invention

In order to confirm that the compounds of the present invention have selective proliferation inhibitory activity against hepatic stellate cells, the activity against hepatic astrocytic HSC-T6, HT 1080 and Rat2 cells was measured and compared. Was inoculated with 1000 cells per well. After 24 hours of inoculation, the compounds of the present invention (number 100 in Table 6) dissolved in DMSO were treated at different concentrations and fixed with formalin solution after 48 hours. Here, after treating and staining Sulfurodamine B, the stained die was extracted with 0.1 M Tris-HCl (pH 8.0), and the absorbance was measured at A520 to quantify the cell number. The cells were cultured at 5% carbon dioxide and 37 ° C., HSC-T6 cells in DMEM + 10% FBS, and the remaining cells in RPMI1640 + 10% FBS.

The test results are shown in FIG. 2. As can be seen in Figure 2, the compound of the present invention was shown to show a relatively high proliferation inhibitory activity against hepatic astrocytic HSC-T6 (-●-) compared to other cells. Therefore, it can be confirmed that the compound of the present invention has selective growth inhibitory activity against hepatic stellate cells.

Example 13: Confirmation of Induction of Apoptosis in Hepatic Astrocytes of the Compound of the Present Invention

Compounds of the present invention (No. 100 in Table 6) dissolved in DMSO in HSC T6 cells were treated by concentration, and total DNA was extracted after 24 hours. Apoptosis was confirmed by measuring DNA fragmentation. The extracted DNA was developed on a 1.2% agarose gel, stained with EtBr, and DNA fragments were observed under UV, and the results are shown in FIG. 3. As shown in FIG. 3, it can be seen that as the concentration of the compound of the present invention increases, more DNA fragments are generated. Thus, it can be seen that the compounds of the present invention induce apoptosis of HSC T65 cells.

Example 14 Identification of Therapeutic Effects of Liver Cirrhosis on Compounds of the Invention

Liver cirrhosis was induced by suturing the bile ducts of whistar rats weighing about 220 g. Here, the compound of the present invention dissolved in DMSO (No. 100 in Table 6) was intraperitoneally injected at a concentration of 50 mg / kg for 3 weeks daily. As a control, DMSO injection was used after the bile duct closure. As a normal control, rats with sham surgery without bile duct closure were used. Hepatic collagen was quantified by quantifying the amount of hydroxyproline in liver tissue, and bilirubin levels and AST and ALT were analyzed by blood autoanalyzers of Korea Biochemistry Lab.

The results are shown in Table 8 below.

As can be seen in Table 8, it can be confirmed that liver hardening was induced by the bile duct suture by increasing the level of hydroxyproline, bilirubin, AST and ALT, compared to the case without the bile duct suture. In this case, when cirrhosis is induced, it can be seen that the increase in hydroxyproline, bilirubin, AST, and ALT levels is alleviated when the compound of the present invention is not treated. Thus, the compounds of the present invention can be said to have therapeutic activity against liver hardening.

Example 15 Determination of Synovial Fibroblast Proliferation Inhibitory Activity of Compounds of the Invention

Synovial fibroblasts extracted from rheumatoid patients cultured in an animal cell incubator at 37 ° C. in DMEM medium containing 10% FBS while supplying 5% carbon dioxide were inoculated with 5000 cells per well in a 24-well culture dish, followed by further culture. The three wells were liver-added with formalin, fixed for 30 minutes, washed with water and dried, and the rest of the wells were incubated by treating the compounds of the present invention (number 100 in Table 6) at concentrations of n = 3 for 48 hours, or Alternatively, after 48 hours of continuous incubation without compound treatment, the same amount of formalin solution was added to each well for 30 minutes, washed with water, and dried.

Formalin-fixed cells were stained with a solution of sulfodadrome B in a conventional manner, and then stained with sulfuric acid adsorbed on the cells with 0.1 M Tris-HCl (pH 8.0) solution, and the absorbance was measured at 520 nM. Relative cell numbers in the wells were quantified. The cell number (i.e., absorbance) in the wells immediately before the compound was treated as the reference point (0%), and the cell number (i.e., absorbance) of the wells cultured for 48 hours without treatment of the compound was 100%. In the absence of cells (i.e., absorbance) at -100%, the cell number (i.e., absorbance) when the compound of the present invention was treated at each concentration for 48 hours was calculated in proportion to the% at each compound concentration. Cell viability was obtained.

As can be seen in Figure 4, the compound of the present invention has an inhibitory activity against the proliferation of synovial fibroblasts, this proliferation inhibitory activity was shown to increase in proportion to the concentration. Therefore, the compound of the present invention can be said to have a therapeutic effect against rheumatism caused by abnormal proliferation of synovial fibroblasts.

Example 16: Determination of Effect of Compounds of the Invention on MMP-1 mRNA

When synovial fibroblasts extracted from rheumatism patients were cultured in DMEM medium containing 10% FBS and 5% CO ₂ in a 10 cm diameter dish, the cell density of the culture dish was about 50%. The compound of the present invention dissolved in DMSO was treated for 24 hours at a concentration of 10 uM or left untreated for 24 hours without treatment with a control compound, and then the total RNA in the cells in the culture dish was purified. Purification was performed using Triazol Reagent (Catalog No .: 15596-026) purchased from GIBCO BRL, and the purification method was separated according to the "Instruction for RNA Isolation" method that GIBCO BRL provided Triazol Reagent with the sales.

10 ug of purified total RNA was electrophoresed on formaldehyde-agarose gel, followed by transfer of mRNA to the nitran membrane. MMP-1 probes were labeled with ³² P radioisotopes using kits purchased from Boehringer Ingelheim using MMP-1 c-DNA fragments. The mRNA amount of MMP-1 was quantified by conventional Northern blotting using the probe thus prepared.

The results are shown in FIG. 5. As shown in Figure 5, when comparing the compound of the present invention treated (10 uM) and untreated (0 uM), the effect of the compound of the present invention to reduce the amount of mRNA of MMP-1 It can be seen that it has. Therefore, the compound of the present invention can be said to have therapeutic activity against rheumatism caused by an increase in the amount of mRNA of MMP-1.

The novel furopyrimidine compounds of the present invention have an excellent inhibitory activity against tyrosine kinase activity of DDR2 protein and are applied as a therapeutic agent for various diseases caused by tyrosine kinase activity of DDR2 by the tyrosine kinase inhibitory activity of such DDR2 protein. It is possible and particularly useful as a therapeutic agent for cirrhosis, rheumatism or cancer.

Figure 1 is an electrophoresis picture showing the tyrosine phosphorylation inhibitory activity of DDR2 protein induced by collagen type 1 in hepatic astrocytic HSC T6 by the compound of the present invention.

Figure 2 is a graph showing the survival rate of the compound treated concentration of cells treated with the compound of the present invention.

Figure 3 is an electrophoresis picture showing the degree of fragmentation of HSC T6 cells treated with a compound of the present invention by treatment concentration of the compound.

Figure 4 is a graph showing the concentration of the compound treated the survival rate of synovial fibroblasts treated with the compound of the present invention.

Figure 5 is an electrophoresis picture showing the comparison of the amount of MMP-1 mRNA in synovial fibroblasts treated with and without the compound of the present invention.

Claims (9)

Furopyrimidine compounds, precursors thereof, and pharmaceutically acceptable salts thereof, as defined by Formula 1: [Formula 1] In the above formula, Z is O, S or NH, n is an integer from 0 to 4, R ₁ is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, amidine group, a haloalkyl group of C alkyl group of _{1 -C 4, C 1 -C 4} , C 1 - a C ₇ alkoxy group, C ₁ -C ₄ alkyl group, C ₁ alkylthio -C _4, C ₁ -C ₄ alkyl amide group, C ₁ -C ₄ acyl group, C ₁ -C ₄ Isiloxy group, C ₁ -C ₄ alkylsulfonamide group, C ₁ -C ₄ alkylsulfonate group, imidic acid C ₁ -C ₄ alkyl ester, thiimidic acid C ₁ -C ₄ alkyl ester, hydroxyna amino, Mo morpholine, acetate, acetamide, methanesulfonamide group is substituted with a methyl group, a halo-C ₁ -C ₄ alkyl group, an alkoxy group of C ₁ -C _4, or halogen-substituted one by groups substituted benzyloxy independently disubstituted Case, The A ring represents benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, cyclohexyl, piperidine or morpholine, R ₂ is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, an alkyl group of _{C 1 -C 5, C 1 -C} 5 haloalkyl group of the alkyl ester, a phenyl group, a halogen Represents a substituted phenyl group, a phenyl group substituted with a C ₁ -C ₄ alkoxy group or a phenyl group substituted with a C ₁ -C ₄ haloalkoxy group, R is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, amidine group, a haloalkyl group of C ₁ -C ₄ alkyl group, a C ₁ -C _4, C ₁ -C ₇ alkoxy, C ₁ -C ₄ alkylamino group import, C ₁ -C ₄ in the alkyl group, C ₁ -C ₄ alkyl amide group, an acylamino group of the C ₁ -C _4, a C ₁ -C ₄ acyl oxy group, or represents the groups when substituted by one alkyl sulfonamide of the C ₁ -C _4, or disubstituted independently. Furopyrimidine compounds, precursors thereof, and pharmaceutically acceptable salts thereof, as defined by Formula 2: [Formula 2] In the above formula, Z is O, S or NH, X is O, S or NH, n is an integer from 0 to 4, n 'is an integer from 0 to 4, R ₁ and R ₃ are the same as or different from each other independently, and are an alkyl group of hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH ₂ , CSNH ₂ , amidine, C ₁ -C ₄ , alkyl amide of the C ₁ -C ₄ haloalkyl groups, C ₁ -C ₇ alkoxy group, C ₁ -C ₄ alkylamino group import, of C ₁ -C ₄ alkylthio, C ₁ -C ₄ group, C ₁ of the -C ₄ acylamino group, a C ₁ -C ₄ Isil oxy group, C ₁ -C ₄ alkyl sulfonamide group, a C ₁ -C ₄ alkyl sulfonate group, an already diksan C ₁ -C ₄ alkyl esters, thio already diksan C ₁ -C ₄ alkyl ester, hydroxy or amino, morpholine base, acetate, acetamide, methanesulfonamide group is substituted with a methyl group, an alkoxy group, a halogen of the haloalkyl group of C ₁ -C _4, C ₁ -C ₄ The case where it is mono-substituted or independently substituted by this substituted benzyloxy group is shown, The A rings are each independently benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, cyclohexyl, piperidine or morpholine Ring represents pyrrole, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, piperidine or morpholine, R ₂ is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, an alkyl group of _{C 1 -C 5, C 1 -C} 5 haloalkyl group of the alkyl ester, a phenyl group, a halogen a substituted phenyl group, C ₁ -C ₄ alkoxy refers to an alkoxy group of the halo-substituted phenyl group or a C ₁ -C ₄ group is substituted phenyl. Furopyrimidine compounds, precursors thereof, and pharmaceutically acceptable salts thereof, as defined by Formula 3: [Formula 3] In the above formula, Z is O, S or NH, R ₁ is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, amidine group, a haloalkyl group of C alkyl group of _{1 -C 4, C 1 -C 4} , C 1 - the C ₄ alkoxy group, C ₁ -C ₄ alkyl group, C ₁ alkylthio -C _4, C ₁ -C ₄ alkyl amide group, C ₁ -C ₄ acyl group, C ₁ -C ₄ Isiloxy group, C ₁ -C ₄ alkylsulfonamide group, C ₁ -C ₄ alkylsulfonate group, imidic acid C ₁ -C ₄ alkyl ester, thiimidic acid C ₁ -C ₄ alkyl ester, hydroxyna amino, Mo morpholine, acetate, acetamide, methanesulfonamide group is substituted with a methyl group, a halo-C ₁ -C ₄ alkyl group, an alkoxy group of C ₁ -C _4, or halogen-substituted one by groups substituted benzyloxy independently disubstituted Case, The A ring represents benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, cyclohexyl, piperidine or morpholine, R ₂ is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, an alkyl group of _{C 1 -C 5, C 1 -C} 5 haloalkyl group of the alkyl ester, a phenyl group, a halogen Represents a substituted phenyl group, a phenyl group substituted with a C ₁ -C ₄ alkoxy group or a phenyl group substituted with a C ₁ -C ₄ haloalkoxy group, R is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, amidine group, a haloalkyl group of C ₁ -C ₄ alkyl group, a C ₁ -C _4, C ₁ -C ₇ alkoxy, C ₁ -C ₄ alkylamino group import, C ₁ -C ₄ in the alkyl group, C ₁ -C ₄ alkyl amide group, an acylamino group of the C ₁ -C _4, a C ₁ -C ₄ acyl oxy group, or represents the groups when substituted by one alkyl sulfonamide of the C ₁ -C _4, or disubstituted independently. Furopyrimidine compounds, precursors thereof, and pharmaceutically acceptable salts thereof, as defined by Formula VI or VII: [Formula VI] In the above formula, R ₂ is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, an alkyl group of _{C 1 -C 5, C 1 -C} 5 haloalkyl group of the alkyl ester, a phenyl group, a halogen Represents a substituted phenyl group, a phenyl group substituted with a C ₁ -C ₄ alkoxy group or a phenyl group substituted with a C ₁ -C ₄ haloalkoxy group, R 'is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, amidine group, a haloalkyl group of C alkyl group of _{1 -C 4, C 1 -C 4} , C 1 - a C ₇ alkoxy group, C ₁ -C ₄ alkyl group, C ₁ alkylthio -C _4, C ₁ -C ₄ alkyl amide group, C ₁ -C ₄ acyl group, C ₁ -C ₄ substituted by an acyloxy group or an alkyl sulfonamide of the C ₁ -C ₄ days or indicates when disubstituted independently. A therapeutically effective amount containing at least one selected from the group consisting of novel furopyrimidine derivatives defined by the following Chemical Formulas 1, 2, 3, VI and VII and pharmacologically acceptable salts thereof as an active ingredient Inhibitors of tyrosine kinase activity of DDR2 protein. [Formula 1] [Formula 2] [Formula 3] [Formula VI] [Formula VII] In the above formula, Z is O, S or NH, X is O, S or NH, n is an integer from 0 to 4, n 'is an integer from 0 to 4, R ₁ is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, amidine group, a haloalkyl group of C alkyl group of _{1 -C 4, C 1 -C 4} , C 1 - a C ₇ alkoxy group, C ₁ -C ₄ alkyl group, C ₁ alkylthio -C _4, C ₁ -C ₄ alkyl amide group, C ₁ -C ₄ acyl group, C ₁ -C ₄ Isiloxy group, C ₁ -C ₄ alkylsulfonamide group, C ₁ -C ₄ alkylsulfonate group, imidic acid C ₁ -C ₄ alkyl ester, thiimidic acid C ₁ -C ₄ alkyl ester, hydroxyna amino, Mo morpholine, acetate, acetamide, methanesulfonamide group is substituted with a methyl group, a halo-C ₁ -C ₄ alkyl group, an alkoxy group of C ₁ -C _4, a substituted one by a halogen-substituted benzyloxycarbonyl, or independently disubstituted Case, The A rings are each independently benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, cyclohexyl, piperidine or morpholine Ring represents pyrrole, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, piperidine or morpholine, R ₂ is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, an alkyl group of _{C 1 -C 5, C 1 -C} 5 haloalkyl group of the alkyl ester, a phenyl group, a halogen This substituted phenyl group, a phenyl group substituted with a C ₁ -C ₄ alkoxy group or a phenyl group substituted with a C ₁ -C ₄ haloalkoxy group, R ³ is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, amidine group, a haloalkyl group of C alkyl group of _{1 -C 4, C 1 -C 4} , C 1 - a C ₇ alkoxy group, C ₁ -C ₄ alkyl group, C ₁ alkylthio -C _4, C ₁ -C ₄ alkyl amide group, C ₁ -C ₄ acyl group, C ₁ -C ₄ Isiloxy group, C ₁ -C ₄ alkylsulfonamide group, C ₁ -C ₄ alkylsulfonate group, imidic acid C ₁ -C ₄ alkyl ester, thiimidic acid C ₁ -C ₄ alkyl ester, hydroxyna amino, Mo morpholine, acetate, acetamide, methanesulfonamide group is substituted with a methyl group, a halo-C ₁ -C ₄ alkyl group, an alkoxy group of C ₁ -C _4, a substituted one by a halogen-substituted benzyloxycarbonyl, or independently disubstituted Case, R is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, amidine group, a haloalkyl group of C ₁ -C ₄ alkyl group, a C ₁ -C _4, C ₁ -C ₇ alkoxy, C ₁ -C ₄ alkylamino group import, C ₁ -C ₄ in the alkyl group, C ₁ -C ₄ alkyl amide group, an acylamino group of the C ₁ -C _4, a C ₁ -C ₄ acyl Mono- or independently substituted by an oxy or C ₁ -C ₄ alkylsulfonamide group. R 'is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, amidine group, a haloalkyl group of C alkyl group of _{1 -C 4, C 1 -C 4} , C 1 - a C ₇ alkoxy group, C ₁ -C ₄ alkyl group, C ₁ alkylthio -C _4, C ₁ -C ₄ alkyl amide group, C ₁ -C ₄ acyl group, C ₁ -C ₄ substituted by an acyloxy group or an alkyl sulfonamide of the C ₁ -C ₄ days or indicates when disubstituted independently. The inhibitor of tyrosine kinase activity of DDR2 protein according to claim 5, further comprising a pharmacologically acceptable carrier or excipient. A therapeutically effective amount containing at least one selected from the group consisting of novel furopyrimidine derivatives defined by the following Chemical Formulas 1, 2, 3, VI and VII and pharmacologically acceptable salts thereof as an active ingredient A therapeutic agent for diseases caused by tyrosine kinase activity of DDR2 protein. [Formula 1] [Formula 2] [Formula 3] [Formula VI] [Formula VII] In the above formula, Z is O, S or NH, X is O, S or NH, n is an integer from 0 to 4, n 'is an integer from 0 to 4, R ₁ is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, amidine group, a haloalkyl group of C alkyl group of _{1 -C 4, C 1 -C 4} , C 1 - a C ₇ alkoxy group, C ₁ -C ₄ alkyl group, C ₁ alkylthio -C _4, C ₁ -C ₄ alkyl amide group, C ₁ -C ₄ acyl group, C ₁ -C ₄ Isiloxy group, C ₁ -C ₄ alkylsulfonamide group, C ₁ -C ₄ alkylsulfonate group, imidic acid C ₁ -C ₄ alkyl ester, thiimidic acid C ₁ -C ₄ alkyl ester, hydroxyna amino, Mo morpholine, acetate, acetamide, methanesulfonamide group is substituted with a methyl group, a halo-C ₁ -C ₄ alkyl group, an alkoxy group of C ₁ -C _4, a substituted one by a halogen-substituted benzyloxycarbonyl, or independently disubstituted Case, The A rings are each independently benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, cyclohexyl, piperidine or morpholine Ring represents pyrrole, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, piperidine or morpholine, R ₂ is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, an alkyl group of _{C 1 -C 5, C 1 -C} 5 haloalkyl group of the alkyl ester, a phenyl group, a halogen This substituted phenyl group, a phenyl group substituted with a C ₁ -C ₄ alkoxy group or a phenyl group substituted with a C ₁ -C ₄ haloalkoxy group, R ³ is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, amidine group, a haloalkyl group of C alkyl group of _{1 -C 4, C 1 -C 4} , C 1 - a C ₇ alkoxy group, C ₁ -C ₄ alkyl group, C ₁ alkylthio -C _4, C ₁ -C ₄ alkyl amide group, C ₁ -C ₄ acyl group, C ₁ -C ₄ Isiloxy group, C ₁ -C ₄ alkylsulfonamide group, C ₁ -C ₄ alkylsulfonate group, imidic acid C ₁ -C ₄ alkyl ester, thiimidic acid C ₁ -C ₄ alkyl ester, hydroxyna amino, Mo morpholine, acetate, acetamide, methanesulfonamide group is substituted with a methyl group, a halo-C ₁ -C ₄ alkyl group, an alkoxy group of C ₁ -C _4, a substituted one by a halogen-substituted benzyloxycarbonyl, or independently disubstituted Case, R is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, amidine group, a haloalkyl group of C ₁ -C ₄ alkyl group, a C ₁ -C _4, C ₁ -C ₇ alkoxy, C ₁ -C ₄ alkylamino group import, C ₁ -C ₄ in the alkyl group, C ₁ -C ₄ alkyl amide group, an acylamino group of the C ₁ -C _4, a C ₁ -C ₄ acyl Mono- or independently substituted by an oxy or C ₁ -C ₄ alkylsulfonamide group. R 'is hydrogen, halogen, cyano, nitro, hydroxy, amino, CO ₂ H, CONH _2, CSNH _2, amidine group, a haloalkyl group of C alkyl group of _{1 -C 4, C 1 -C 4} , C 1 - a C ₇ alkoxy group, C ₁ -C ₄ alkyl group, C ₁ alkylthio -C _4, C ₁ -C ₄ alkyl amide group, C ₁ -C ₄ acyl group, C ₁ -C ₄ substituted by an acyloxy group or an alkyl sulfonamide of the C ₁ -C ₄ days or indicates when disubstituted independently. 8. The therapeutic agent according to claim 7, wherein the disease caused by DDR2 tyrosine kinase activity is cirrhosis, rheumatism or tumor. The therapeutic agent of claim 7 or 8 further comprising a pharmacologically acceptable carrier or excipient.