# Patent

## Title
Arylamino purine derivatives, preparation method and pharmaceutical use thereof

## Abstract

Arylamino purine derivatives represented by formula I and their preparation method are disclosed, wherein each substituent is defined as in the description. The derivatives have an inhibitory effect on non-small cell lung cancer with deletion mutation of exon 19 or L858R point mutation of exon 21 in epidermal growth factor receptor (EGFR).

## Background
TECHNICAL FIELD
The present invention relates to the organically synthesized pharmaceutical field and in particular to arylamino purine derivatives, preparation method and pharmaceutical use thereof.
BACKGROUNDS
Malignant tumor is one of the most severe diseases threatening the human physical health, and has become the second leading cause of death, right behind the cardiovascular disease.
Moreover, the most recent statistical data indicate that the incidence rate and the death rate of malignant tumor are rising up year by year all over the world, especially in the developing countries.
The chemotherapy is the most important treatment means for malignant tumor besides operation and radiotherapy. The traditional chemotherapeutics mainly act on DNA, RNA, microtubule protein and the like, which are the common parts involving the life and death of all cells, and therefore have low selectivity and high toxicity. The targeted medicines act on the key molecules in the tumor cell, which regulate the cell growth and proliferation and are quite different from the normal cell, and their signal transduction pathways. They have the advantages such as a high selectivity on the tumor cells and low toxicity to normal tissues, and therefore become a hot point in the study of anti-tumor drug.
In many of molecules regulating signal transduction pathway of the cell, the family of protein kinases is the most important signal transduction molecule. It is found in the study that the occurrence and development of many tumors are relevant to the gene abnormality or excess activation of protein kinase. Therefore, protein kinases have become the most important anti-tumor treatment target. Tyrosine or serine/threonine protein kinase such as EGFR (Epidermal Growth Factor Receptor), VEGFR (Vascular Endothelial Growth Factor Receptor), PDGFR (Platelet Derived Growth Factor Receptor), c-Kit, c-SRC, MET, BTK, ALK, Abl, and FLT3 are most important among the members of the family of protein kinases, and have been listed as oncogene or oncoprotein.
Currently, more than 10 small molecule inhibitors targeting these tyrosine and serine/threonine protein kinases have been applied to clinical tumor treatment. The typical example includes: two EGFR inhibitors, i.e., Gefitinib and Erlotinib, mainly useful for the treatment of non-small cell lung carcinoma; two new-vessel inhibitors (their main targets are VEGFR, PDGFR and the like), i.e., Sunitinib and Sorafenib, useful for the anti-new-vessels treatment in the solid tumor; and Bcr-Abl kinase inhibitor Imatinib, mainly useful for the treatment of chronic myelocytic leukemia with positive Philadelphia chromosome.
However, these current antitumor drugs target a single or a very small number of kinase oncogenes or oncoproteins and therefore have the disadvantages such as low efficiency and being prone to develop the drug resistance. The typical example includes EGFR kinase inhibitors Gefitinib and Erlotinib. These two drugs have good effects on only 10-20% of patients with non-small cell lung carcinoma. It is found in the study on its mechanism of action that Gefitinib and Erlotinib belong to the selective EGFR kinase inhibitor, and are most sensitive to the patients having deletion mutation of exon 19 or L858R point mutation of exon 21 in EGFR (deletion mutation of exon 19 and L858R point mutation of exon 21 in EGFR are collectively referred to as EGFR sensitive mutation). Even if these patients with EGFR sensitive mutation are treated with Gefitinib and Erlotinib, most of them have developed the drug resistance after 6-9 months. It is found that there are many reasons for the development of the drug resistance, which mainly include (1) a second mutation occurs based on EGFR sensitive mutation, i.e., T790M mutation in EGFR; and (2) MET gene (MET is a tyrosine kinase) is subjected to amplification.
Solving the low efficiency and the drug resistance of small molecule kinase inhibitor antitumor drug is not only a hot point in the study but also a task of top priority. Currently, the researchers and scientists are exploring the effective solutions, in which the most promising solutions mainly include: (1) a multiple-kinase inhibitor that simultaneously targets multiple kinases relevant to the occurrence and development of tumor; (2) directly targeting the kinase that has developed the drug-resistant mutation. For example, EGFR is over expressed or abnormally and excessively activated in several tumor tissues of human. If a certain small molecule kinase inhibitor not only can inhibit the activity of EGFR, but also can inhibit the new-vessels or the activity of the oncogene or oncoprotein of the other key kinases regulating the cell growth and proliferation, e.g. kinases such as c-Kit, c-SRC, MET, BTK, ALK, Abl and FLT3, it can improve the efficiency of tumor treatment and reduce the occurrence rate of the drug resistance. For example, as stated hereinbefore, after EGFR inhibitors Gefitinib and Erlotinib are used, EGFR itself is prone to experience a second mutation based on the original mutation (i.e., EGFR sensitive mutation), i.e., T790M mutation in EGFR. The occurrence of this second mutation is one of the main reasons causing EGFR inhibitors Gefitinib and Erlotinib to be ineffective. Therefore, the research and development of an EGFR kinase inhibitor that directly targets the drug resistant mutation (i.e. T790M mutation) is a direct approach of overcoming the drug resistance of this tumor.
SUMMARY OF INVENTION
A technical solution to be solved in the present invention is to provide an arylamino purine derivative represented by formula (I).




R1 represents —H, —CmH(2m+1), C3-C7cycloalkyl, —CmH(2m+1) substituted by C3-C7cycloalkyl, C3-C7cycloalkyl substituted by —CmH(2m+1), heterocyclyl containing 3-8 carbon atoms, amino substituted by heterocyclyl containing 3-8 carbon atoms, aryl containing 6-8 carbon atoms, or heteroaryl containing 6-8 carbon atoms; said heterocyclyl contains 1-3 heteroatoms selected from N, O and S; said heteroaryl contains 1-3 heteroatoms selected from N, O and S;

R2 represents —H, —NH2, —OH, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1), —NHCmH(2m+1), aryloxy containing 6-12 carbon atoms, or arylamino containing 6-12 carbon atoms;

R3 represents C3-C7cycloalkyl, C3-C7cycloalkyl substituted by —CmH(2m+1), aryl containing 6-80 carbon atoms, or heteroaryl containing 6-80 carbon atoms; said heteroaryl contains 1-15 heteroatoms selected from N, O and S;

R4-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —OCmH(2m+1),





Said aryl containing 6-8 carbon atoms may be an aryl only containing C and H, or may be an aryl having a substituent, wherein said substituent can be —NH2, —OH, —F, —Cl, —Br, —CF3, —CyH(2y+1), —OCyH(2y+1) and/or —NHCyH(2y+1), where y=1-5. Said aryl having a substituent can contain 0-8 nitrogen or oxygen heteroatoms; and
Said heteroaryl may be a heteroaryl only containing C, H, N, O or S, or may be a heteroaryl having a substituent, wherein said heteroaryl can be —NH2, —OH, —F, —Cl, —Br and/or —CF3.
Preferably, the arylamino purine derivative is represented by formula (I), wherein
R1 represents —H, —CmH(2m+1), C3-C7cycloalkyl substituted by —CmH(2m+1), —CmH(2m+1) substituted by C3-C7cycloalkyl, or C3-C7cycloalkyl;
R2 represents —H, —NH2, —OH, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1), or —NHCmH(2m+1);
R3 represents




pyrimidinyl, halopyrimidinyl, C3-C7cycloalkyl, or C3-C7cycloalkyl substituted by —CmH(2m+1);

R4-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —OCmH(2m+1),





R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CN, —CmH(2m+1), —OCmH(2m+1),





R14 represents aryl or heteroaryl containing 6-10 carbon atoms; said heteroaryl contains 1-3 heteroatoms selected from N, O and S; m=1-8; and

n=0-4.

Preferably, the arylamino purine derivative is represented by formula (I), wherein
R1 represents —H, —CmH(2m+1), C3-C7cycloalkyl substituted by —CmH(2m+1), —CmH(2m+1) substituted by C3-C7cycloalkyl, or C3-C7cycloalkyl;
R2 represents —H, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1), or —NHCmH(2m+1);
R3 represents




halopyrimidin-3-yl, C3-C7cycloalkyl, or C3-C7cycloalkyl substituted by —CmH(2m+1);

R4-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCmH(2m+1),





R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —CN, —CmH(2m+1), —OCmH(2m+1),





and

m=1-8.

Most preferably, the arylamino purine derivative is represented by formula (I), wherein
R1 represents —H, —CmH(2m+1), —CmH(2m+1) substituted by C3-C2cycloalkyl, or C3-C2cycloalkyl;
R2 represents —H, —CmH(2m+1), —OCmH(2m+1), —NHCmH(2m+1);
R3 represents




halopyrimidin-3-yl, or





R4 represents





R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, or —OCmH(2m+1);

R9-R13 respectively represent —H, —F, —Cl, —Br, CF3, —CmH(2m+1), —OCmH(2m+1),





m=1-8.

Further, the arylamino purine derivative is represented by formula (II), wherein




R1 represents —H, —CmH(2m+1), —C6H5, —CmH(2m+1) substituted by C3-C2cycloalkyl, C3-C2cycloalkyl, or C3-C2cycloalkyl substituted by —CmH(2m+1);

R2 represents —H, —NH2, —OH, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1), —NHCmH(2m+1), aryloxy containing 6-12 carbon atoms, or arylamino containing 6-12 carbon atoms;

R3 represents C3-C2cycloalkyl, C3-C2cycloalkyl substituted by —CmH(2m+1), aryl containing 6-80 carbon atoms or heteroaryl containing 6-80 carbon atoms; said heteroaryl contains 1-15 heteroatoms selected from N, O and S;

R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —OCmH(2m+1),





R15 represents





m=1-8; and

n=0-4.

Preferably, the arylamino purine derivative is represented by formula (II), wherein
R1 represents —H, —CmH(2m+1), —CmH(2m+1) substituted by C3-C7cycloalkyl, or C3-C7cycloalkyl;
R2 represents —H, NH2, OH, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1), or —NHCmH(2m+1);
R3 represents




pyrimidinyl, halopyrimidinyl, C3-C7cycloalkyl, or C3-C7cycloalkyl substituted by —CmH(2m+1);

R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCmH(2m+1),





R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —CN, —CmH(2m+1),





R14 represents aryl or heteroaryl containing 6-10 carbon atoms; said heteroaryl contains 1-3 heteroatoms selected from N, O and S;

R15 represents





m=1-8; and

n=0-4.

Preferably, the arylamino purine derivative is represented by formula (II), wherein
R1 represents —H, —CmH(2m+1), or C3-C7cycloalkyl;
R2 represents —H, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1), or —NHCmH(2m+1);
R3 represents




halopyrimidin-3-yl, C3-C7cycloalkyl, or C3-C7cycloalkyl substituted by —CmH(2m+1);

R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —OCmH(2m+1),





R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —CN, —CmH(2m+1), —OCmH(2m+1),





R15 represents





and

m=1-8.

Preferably, the arylamino purine derivative is represented by formula (II), wherein
R1 represents —H, —CmH(2m+1),




R2 represents —H;

R3 represents





R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, or —OCmH(2m+1);

R9-R13 respectively represent —H, —F, —Cl, —Br, CF3, —CmH(2m+1), —OCmH(2m+1),





R15 represents





and

m=1-8.

Most preferably, the arylamino purine derivative is represented by formula (II), wherein
R1 represents —H, —CmH(2m+1),




R2 represents —H:

R3 represents





R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, or —OCmH(2m+1);

R8-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1),





R15 represents





and

m=1-8.

Further, the arylamino purine derivative is represented by formula (III), wherein




R1 represents —H, —CmH(2m+1), —C6H5, —CmH(2m+1) substituted by C3-C7cycloalkyl, C3-C7cycloalkyl, or C3-C2cycloalkyl substituted by —CmH(2m+1);

R2 represents —H, —NH2, —OH, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1), —NHCmH(2m+1), aryloxy containing 6-12 carbon atoms, or arylamino containing 6-12 carbon atoms.

R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —OCmH(2m+1),





R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CN, —CmH(2m+1), —OCmH(2m+1),





R14 represents aryl or heteroaryl containing 6-10 carbon atoms; said heteroaryl contains 1-3 heteroatoms selected from N, O and S;

m=1-8; and

n=0-4.

Preferably, the arylamino purine derivative is represented by formula (III), wherein
R1 represents —H, —CmH(2m+1), C3-C7cycloalkyl, or C3-C7cycloalkyl substituted by —CmH(2m+1);
R2 represents —H, —NH2, —OH, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1), or —NHCmH(2m+1);
R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —OCmH(2m+1),




R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CN, —CmH(2m+1), —OCmH(2m+1),





R14 represents aryl or heteroaryl containing 6-10 carbon atoms; said heteroaryl contains 1-3 heteroatoms selected from N, O and S; m=1-8; and

n=0-4.

Preferably, the arylamino purine derivative is represented by formula (Ill), wherein
R1 represents —H, —CmH(2m+1), or C3-C7cycloalkyl;
R2 represents —H, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1), or —NHCmH(2m+1);
R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, or —OCmH(2m+1);
R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —CN, —CmH(2m+1), —OCmH(2m+1),




m=1-8.

Preferably, the arylamino purine derivative is represented by formula (III), wherein
R1 represents —H, —CmH(2m+1),




R2 represents —H, —CmH(2m+1), —CmH(2m+1), or —NHCmH(2m+1);

R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, or —OCmH(2m+1);

R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CmH(2m+1), —OCmH(2m+1),





and

m=1-8.

Preferably, the arylamino purine derivative is represented by formula (III), wherein
R1 represents —H, —CmH(2m+1),




R2 represents —H; R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, or —OCmH(2m+1);

R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1),





m=1-8.

Most preferably, the arylamino purine derivative is represented by formula (III), wherein
R1 represents —H, —CmH(2m+1),




R2 represents —H; R5-R8 represent —H;

R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1),





m=1-8.

Further, the arylamino purine derivative is represented by formula (IV), wherein




R1 represents —H, —C11H(2m+1), —C6H5, —CmH(2m+1) substituted by C3-C7cycloalkyl, C3-C7cycloalkyl, or C3-C2cycloalkyl substituted by —CmH(2m+1);

R2 represents —H, —NH2, —OH, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1), —NHCmH(2m+1), aryloxy containing 6-12 carbon atoms, or arylamino containing 6-12 carbon atoms;

R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —OCmH(2m+1),





R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CN, —CmH(2m+1), —OCmH(2m+1),





R14 represents aryl or heteroaryl containing 6-10 carbon atoms; said heteroaryl contains 1-3 heteroatoms selected from N, O and S; m=1-8; and n=0-4.

Preferably, the arylamino purine derivative is represented by formula (IV), wherein
R1 represents —H, —CmH(2m+1), —CmH(2m+1) substituted by C3-C7cycloalkyl, C3-C7cycloalkyl, or C3-C2cycloalkyl substituted by —CmH(2m+1);
R2 represents —H, —NH2, —OH, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1), or —NHCmH(2m+1);
R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —OCmH(2m+1),




R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CN, —CmH(2m+1), —OCmH(2m+1),





R14 represents aryl or heteroaryl containing 6-10 carbon atoms; said heteroaryl contains 1-3 heteroatoms selected from N, O and S; m=1-8; and

n=0-4.

Preferably, the arylamino purine derivative is represented by formula (IV), wherein
R1 represents —H, —CmH(2m+1), C3-C2cycloalkyl, or C3-C7cycloalkyl substituted by —CmH(2m+1);
R2 represents —H, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1), or —NHCmH(2m+1);
R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —OCmH(2m+1),




R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —CN, —CmH(2m+1), —OCmH(2m+1),





and

m=1-8.

Preferably, the arylamino purine derivative is represented by formula (IV), wherein
R1 represents —H, —CmH(2m+1), or C3-C7cycloalkyl;
R2 represents —H, —F, —Cl, —Br, —CmH(2m+1), —OCmH(2m+1), or —NHCmH(2m+1);
R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, or —OCmH(2m+1);
R9-R13 respectively represent —H, —F, —Cl, —Br, CF3, —CmH(2m+1), —OCmH(2m+1),




and

m=1-8.

Preferably, the arylamino purine derivative is represented by formula (IV), wherein
R1 represents —H, —CmH(2m+1),




R2 represents —H, —CmH(2m+1), —OCmH(2m+1), —NHCmH(2m+1);

R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, or —CmH(2m+1);

R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CmH(2m+1), —OCmH(2m+1),





and

m=1-8.

Preferably, the arylamino purine derivative is represented by formula (IV), wherein
R1 represents —H, —CmH(2m+1),




R2 represents —H;

R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, or —OCmH(2m+1);

R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1),





and

m=1-8.

Most preferably, the arylamino purine derivative is represented by formula (IV), wherein
R1 represents —H, —CmH(2m+1),




R2 represents —H;

R5-R8 represent H;

R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1), or





m=1-8.

Further, the arylamino purine derivative is represented by formula (V), wherein




R1 represents —H, —CmH(2m+1), —C6H5, C3-C7cycloalkyl, —CmH(2m+1) substituted by C3-C7cycloalkyl, or C3-C2cycloalkyl substituted by —CmH(2m+1);

R2 represents —H, —NH2, —OH, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1), —NHCmH(2m+1), aryloxy containing 6-12 carbon atoms, or arylamino containing 6-12 carbon atoms;

R3 represents C3-C7cycloalkyl, C3-C7cycloalkyl substituted by —CmH(2m+1), aryl containing 6-80 carbon atoms or heteroaryl containing 6-80 carbon atoms; said heteroaryl contains 1-15 heteroatoms selected from N, O and S;

R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —OCmH(2m+1),





m=1-8; and

n=0-4.

Preferably, the arylamino purine derivative is represented by formula (V), wherein
R1 represents —H, —CmH(2m+1), C3-C7cycloalkyl, —CmH(2m+1) substituted by C3-C7cycloalkyl, or C3-C2cycloalkyl substituted by —CmH(2m+1);
R2 represents —H, —NH2, —OH, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1), or —NHCmH(2m+1);
R3 represents




pyrimidinyl, halopyrimidinyl, C3-C7cycloalkyl, or C3-C2cycloalkyl substituted by —CmH(2m+1);

R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —OCmH(2m+1),





R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CN, —CmH(2m+1), —OCmH(2m+1),





R14 represents aryl or heteroaryl containing 6-10 carbon atoms; said heteroaryl contains 1-3 heteroatoms selected from N, O and S; m=1-8; and

n=0-4.

Preferably, the arylamino purine derivative is represented by formula (V), wherein
R1 represents —H, —CmH(2m+1), C3-C7cycloalkyl, —CmH(2m+1) substituted by C3-C7cycloalkyl, or C3-C7cycloalkyl substituted by —CmH(2m+1);
R2 represents —H, —F, —Cl, —Br, —CF3, —CmH(2m+1, —OCmH(2m+1), or —NHCmH(2m+1);
R3 represents




halopyrimidin-3-yl, C3-C7cycloalkyl, or C3-C7cycloalkyl substituted by —CmH(2m+1);

R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —OCmH(2m+1),





R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —CN, —CmH(2m+1), —OCmH(2m+1),





and

m=1-8.

Preferably, the arylamino purine derivative is represented by formula (V), wherein
R1 represents —H, —CmH(2m+1), —CmH(2m+1) substituted by C3-C7cycloalkyl, or C3-C7cycloalkyl;
R2 represents —H, —F, —Cl, —Br, —CmH(2m+1), —OCmH(2m+1), or —NHCmH(2m+1);
R3 represents




halopyrimidin-3-yl, or





R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, or —OCmH(2m+1);

R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —CmF(2m+1), —OCmH(2m+1),





and

m=1-8.

Preferably, the arylamino purine derivative is represented by formula (V), wherein
R1 represents —H, —CmH(2m+1),




R2 represents —H, —CmH(2m+1), —OCmH(2m+1), —NHCmH(2m+1);

R3 represents





halopyrimidin-3-yl, or





R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, or —OCmH(2m+1);

R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —CmFl(2m+1), —OCmH(2m+1),





and

m=1-8.

Preferably, the arylamino purine derivative is represented by formula (V), wherein
R1 represents —H, —CmH(2m+1),




R2 represents-H;

R3 represents





R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, or —OCmH(2m+1);

R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CmH(2m+1), —OCmH(2m+1),





and

m=1-8.

Most preferably, R1 represents —H, —CmH(2m+1),




R2 represents —H;

R3 represents





R5-R8 represent —H;

R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCmH(2m+1),





and

m=1-8.

Further, the arylamino purine derivative is represented by formula (VI), wherein




R1 represents —H, —CmH(2m+1), —C6H5, C3-C7cycloalkyl, —CmH(2m+1) substituted by C3-C7cycloalkyl, or C3-C2cycloalkyl substituted by —CmH(2m+1);

R2 represents —H, —NH2, —OH, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1), —NHCmH(2m+1), aryloxy containing 6-12 carbon atoms, or arylamino containing 6-12 carbon atoms;

R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —OCmH(2m+1),





R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CN, —CmH(2m+1), —OCmH(2m+1),





R14 represents aryl or heteroaryl containing 6-10 carbon atoms; said heteroaryl contains 1-3 heteroatoms selected from N, O and S; m=1-8; and

n=0-4.

Preferably, the arylamino purine derivative is represented by formula (VI), wherein
R1 represents —H, —CmH(2m+1), C3-C7cycloalkyl, —CmH(2m+1) substituted by C3-C7cycloalkyl, or C3-C2cycloalkyl substituted by —CmH(2m+1);
R2 represents —H, —NH2, —OH, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1), or —NHCmH(2m+1);
R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —OCmH(2m+1),




R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CN, —CmH(2m+1),





R14 represents aryl or heteroaryl containing 6-10 carbon atoms; said heteroaryl contains 1-3 heteroatoms selected from N, O and S; m=1-8; and

n=0-4.

Preferably, the arylamino purine derivative is represented by formula (VI), wherein
R1 represents —H, —CmH(2m+1), C3-C7cycloalkyl, —CmH(2m+1) substituted by C3-C7cycloalkyl, or C3-C7cycloalkyl substituted by —CmH(2m+1);
R2 represents —H, —F, —Cl, —Br, —CF3, —CmH(2m+1), —CmH(2m+1), or —NHCmH(2m+1);
R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —OCmH(2m+1),




R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CN, —CmH(2m+1), —OCmH(2m+1),





m=1-8.

Preferably, the arylamino purine derivative is represented by formula (VI), wherein
R1 represents —H, —CmH(2m+1), —CmH(2m+1) substituted by C3-C7cycloalkyl, or C3-C7cycloalkyl;
R2 represents —H, —F, —Cl, —Br, —CmH(2m+1), —OCmH(2m+1), or —NHCmH(2m+1);
R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, or —OCmH(2m+1);
R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CmH(2m+1), —OCmH(2m+1),




and

m=1-8.

Preferably, the arylamino purine derivative is represented by formula (VI), wherein
R1 represents —H, —CmH(2m+1),




R2 represents —H, —CmH(2m+1), —OCmH(2m+1), —NHCmH(2m+1);

R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, or —OCmH(2m+1);

R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CmH(2m+1), —OCmH(2m+1),





and

m=1-8.

Preferably, the arylamino purine derivative is represented by formula (VI), wherein
R1 represents —H, —CmH(2m+1),




R2 represents —H; R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, or —OCmH(2m+1);

R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CmH(2m+1), —OCmH(2m+1),





and

m=1-8.

Most preferably, the arylamino purine derivative is represented by formula (VI), wherein
R1 represents —H, —CmH(2m+1),




R2 represents —H; R5-R8 represent —H;

R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCmH(2m+1),





and

m=1-8.

Preferably, the arylamino purine derivative is represented by formula (VII), wherein




R1 represents —H, —CmH(2m+1), —C6H5, C3-C7cycloalkyl, —CmH(2m+1) substituted by C3-C7cycloalkyl, or C3-C7cycloalkyl substituted by —CmH(2m+1);

R2 represents —H, —NH2, —OH, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1), —NHCmH(2m+1), aryloxy containing 6-12 carbon atoms, or arylamino containing 6-12 carbon atoms;

R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —OCmH(2m+1),





R10-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CN, —CmH(2m+1), —OCmH(2m+1),





R16 represents





R14 represents —CmH(2m+1), aryl or heteroaryl containing 6-10 carbon atoms; said heteroaryl contains 1-3 heteroatoms selected from N, O and S; m=1-8; and

n=0-4.

Preferably, the arylamino purine derivative is represented by formula (VII), wherein
R1 represents —H, —CmH(2m+1), —CmH(2m+1) substituted by C3-C2cycloalkyl, or C3-C2cycloalkyl;
R2 represents —H, —NH2, —OH, —F, —Cl, —Br, —CF3, —CmH(2m+1, —OCmH(2m+1), or —NHCmH(2m+1);
R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —OCmH(2m+1),




R10-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CN, —CmH(2m+1), —OCmH(2m+1),





R14 represents aryl or heteroaryl containing 6-10 carbon atoms; said heteroaryl contains 1-3 heteroatoms selected from N, O and S;

R16 represents





m=1-8; and

n=0-4.

Preferably, the arylamino purine derivative is represented by formula (VII), wherein
R1 represents —H, —CmH(2m+1), —CmH(2m+1) substituted by C3-C2cycloalkyl, or C3-C2cycloalkyl;
R2 represents —H, —F, —Cl, —Br, —CF3, —CmH(2m+1, —OCmH(2m+1), or —NHCmH(2m+1);
R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCmH(2m+1),




R10-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CN, —CmH(2m+1), —OCmH(2m+1),





R16 represents and





m=1-8.

Preferably, the arylamino purine derivative is represented by formula (VII), wherein
R1 represents —H, —CmH(2m+1), —CmH(2m+1) substituted by C3-C7cycloalkyl, or C3-C7cycloalkyl;
R2 represents —H, —CmH(2m+1), —OCmH(2m+1), or —NHCmH(2m+1);
R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, or —OCmH(2m+1);
R10-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CmH(2m+1), —OCmH(2m+1),




R16 represents





and

m=1-8.

Preferably, the arylamino purine derivative is represented by formula (VII), wherein
R1 represents —H, —CmH(2m+1),




R2 represents —H; R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CmH(2m+1), or —OCmH(2m+1);

R10-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CmH(2m+1), —OCmH(2m+1),





R16 represents





and

m=1-8.

Preferably, the arylamino purine derivative is represented by formula (VII), wherein
R1 represents —H, —CmH(2m+1),




R2 represents —H; R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CmH(2m+1), or —OCmH(2m+1);

R10-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CmH(2m+1), or —OCmH(2m+1);

R16 represents





and

m=1-8.

Most preferably, the arylamino purine derivative is represented by formula (VII), wherein
R1 represents —H, —CmH(2m+1),




R2 represents-H;

R5-R8 represent —H;

R10-R13 respectively represent —H, —F, —Cl, or —Br;

R16 represents





and

m=1-8.

Further, the arylamino purine derivative is represented by formula (VIII), wherein




R1 represents —H, —CmH(2m+1), C3-C7cycloalkyl, —CmH(2m+1) substituted by C3-C7cycloalkyl, C3-C7cycloalkyl substituted by —CmH(2m+1), heterocyclyl containing 3-8 carbon atoms, amino substituted by heterocyclyl containing 3-8 carbon atoms, aryl containing 6-8 carbon atoms, or heteroaryl containing 6-8 carbon atoms; said heteroaryl contains 1-3 heteroatoms selected from N, O and S;

R2 represents —H, —NH2, —OH, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1), —NHCmH(2m+1), aryloxy containing 6-12 carbon atoms, or arylamino containing 6-12 carbon atoms;

R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —OCmH(2m+1),





R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CN, —CmH(2m+1), —OCmH(2m+1),





R21 represents





R14 represents aryl or heteroaryl containing 6-10 carbon atoms; said heteroaryl contains 1-3 heteroatoms selected from N, O and S; m=1-8; and

n=0-4.

Preferably, the arylamino purine derivative is represented by formula (VIII), wherein
R1 represents —H, —CmH(2m+1), —CmH(2m+1) substituted by C3-C7cycloalkyl, or C3-C7cycloalkyl;
R2 represents —H, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1), or —NHCmH(2m+1);
R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, or —OCmH(2m+1);
R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —CN, —CmH(2m+1), —OCmH(2m+1), or




R21 represents





m=1-8; and

n=0-4.

Preferably, the arylamino purine derivative is represented by formula (VIII), wherein
R1 represents —H, —CmH(2m+1), —CmH(2m+1) substituted by C3-C7cycloalkyl, or C3-C7cycloalkyl;
R2 represents —H, —CmH(2m+1), —OCmH(2m+1), or —NHCmH(2m+1);
R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, or —OCmH(2m+1);
R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —CmH(2m+1), or —OCmH(2m+1);
R21 represents




m=1-8; and

n=0-4.

Preferably, the arylamino purine derivative is represented by formula (VIII), wherein
R1 represents —H, —CmH(2m+1),




R2 represents —H; R5-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —CmH(2m+1), or —OCmH(2m+1);

R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1);

R21 represents





m=1-8; and

n=0-4.

Most preferably, the arylamino purine derivative is represented by formula (VIII), wherein
R1 represents —H, —CmH(2m+1),




R2 represents —H; R5-R8 represent —H; R9-R13 respectively represent —H, —F, —Cl, —Br, —CF3, —CmH(2m+1), or —OCmH(2m+1);

R21 represents





m=1-8, n=0-2.

Further, the arylamino purine derivative is represented by formula (IX), wherein




R17 represents H, C1-4alkyl, isopropyl, cyclopropyl, or cyclopentyl;

When R18 represents H, R19 represents 4-methylpiperazinyl-1-carbamoyl, 1-methylpiperidinyl-4-carbamoyl, 4-methyl piperazin-1-yl, 4-morpholinyl, 4-methylpiperazin-1-ylmethyl, 4-morpholinylmethyl, 3-(4-methylpiperazin-1-yl)ureido, or 3-(1-methylpiperidin-4-yl)ureido;

When R18 represents fluoro, chloro, bromo, or methoxy, R19 represents 4-methylpiperazinyl-1-carbamoyl, or 1-methylpiperidinyl-4-carbamoyl;

R20 represents hydrogen, halogen, C1-4alkyl, ethynyl, 3-chloro-4-(pyridin-2-yl)methoxy, or 3-chloro-4-(3-fluorophenyl)methoxy.

Preferably, R17 represents H, C1-4alkyl, isopropyl, cyclopropyl, or cyclopentyl;
When R18 represents H, R19 represents 4-methylpiperazinyl-1-carbamoyl, 1-methylpiperidinyl-4-carbamoyl, 4-methyl piperazin-1-yl, 4-morpholinyl, 4-methylpiperazin-1-ylmethyl, 4-morpholinylmethyl, 3-(4-methylpiperazin-1-yl)ureido, or 3-(1-methyl piperidin-4-yl)ureido;

When R18 represents fluoro, chloro, bromo, or methoxy, R19 represents 4-methylpiperazinyl-1-carbamoyl, or 1-methylpiperidinyl-4-carbamoyl;

R20 represents hydrogen, halogen, C1-4alkyl or ethynyl.

Further, the arylamino purine derivative is as below.

    4-(9-isopropyl-8-phenylamino-9H-purin-2-ylamino)-N-(4-methylpiperidin-1-yl)benzamide,
    4-(8-(3-chloro-4-fluorophenylamino)-9-isopropyl-9H-purin-2-ylamino)-N-(4-methylpiperidin-1-yl)benzamide,
    4-(8-(3-acetamidophenylamino)-9-isopropyl-9H-purin-2-ylamino)-N-(4-methylpiperidin-1-yl)benzamide,
    3-fluoro-4-(9-isopropyl-8-phenylamino-9H-purin-2-ylamino)-N-(4-methylpiperazin-1-yl)benzamide,
    3-fluoro-4-(9-isopropyl-8-(3-chloro-4-fluorophenylamino)-9H-purin-2-ylamino)-N-(4-methylpiperazin-1-yl)benzamide,
    3-fluoro-4-(9-isopropyl-8-phenylamino-9H-purin-2-ylamino)-N-(4-methylpiperazin-1-yl)benzamide,
    3-methoxy-4-(9-isopropyl-8-phenylamino-9H-purin-2-ylamino)-N-(4-methylpiperazin-1-yl)benzamide,
    4-(8-(3-chloro-4-fluorophenylamino)-9-isopropyl-9H-purin-2-ylamino)-3-methoxy-N-(1-methylpiperidin-4-yl)benzamide,
    4-(8-phenylamino-9-isopropyl-9H-purin-2-ylamino)-N-(1-methylpiperidin-4-yl)benzamide,
    9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-8-phenylamino-9H-purine,
    8-(3-bromophenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-(3-ethynylphenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-(3-chloro-4-fluorophenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-(3-bromophenylamino)-9-cyclopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-(4-bromophenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-(3-trifluoromethylphenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-(3-methoxyphenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-(4-methoxyphenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-(3-chloro-4-(3-fluorobenzyloxy)phenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-(3-chloro-4-((pyridin-2-yl)methoxy)phenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-(3-(3-(3-chloro-4-fluorophenyl)ureido)phenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-(4-(3-fluorophenylcarbamoyl)phenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-phenylamino-9-cyclopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-phenylamino-9-methyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-phenylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-phenylamino-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-(4-trifluoromethylphenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-(3-acrylylaminophenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-p urine,
    9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-8-(pyridin-3-yl)-9H-purine,
    9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-8-cyclohexyl-9H-purine,
    9-cyclopentyl-6-methyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-8-phenyl-9H-purine,
    9-isopropyl-6-methoxy-2-(4-(4-methylpiperazin-1-yl)phenylamino)-8-phenyl-9H-purine,
    9-isopropyl-6-methylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-8-phenyl-9H-purine,
    9-isopropyl-6-methyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-8-phenyl-9H-purine,
    9-isopropyl-6-methyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-8-phenyl-9H-purine,
    9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-8-(pyridin-3-yl)-9H-purine,
    4-(9-cyclopentylamino-8-(pyridin-3-ylamino)-9H-purin-2-ylamino)-N-(4-methylpiperidin-1-yl)benzamide,
    N-(3-(9-isopropyl-8-phenylamino-9H-purin-2-ylamino)phenyl)acrylamide,
    8-(4-bromophenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-(3-nitrophenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-benzylamino-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-(3-ethynylphenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-(2-fluoro-4-bromophenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-phenylamino-9-cyclopentyl-2-(4-(4-morpholinylmethyl)phenylamino)-9H-purine,
    8-(3-fluorophenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-phenylamino-9-cyclohexyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-(3-hydroxyphenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-(3-chlorophenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-(3-methylphenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-(3,5-dichlorophenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-(2,5-difluorophenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-phenylamino-9-cyclopentyl-2-(4-(2-methoxyethoxy)phenylamino)-9H-purine,
    8-(2,4,5-trichlorophenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-phenylamino-9-cyclopentyl-2-(4-((4-ethylpiperazin-1-yl)methyl)phenylamino)-9H-purine,
    8-phenylamino-9-cyclopentyl-2-(4-((4-methylpiperazin-1-yl)methyl)phenylamino)-9H-purine,
    8-(2,5-dichlorophenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-(2,6-dichlorophenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-(3-bromophenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    8-phenylamino-9-cyclohexylmethyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine,
    9-isopropylamino-2-(4-(2-morpholinoethoxy)phenylamino))-8-phenylamino-9H-purine,
    9-isopropylamino-2-(4-(3-(4-methylpiperazin-1-yl)propylamino)phenylamino)-8-phenylamino-9H-purine,
    8-(6-chloropyrimidin-3-yl)-9-isopropylamino-2-(4-(3-(4-methylpiperazin-1-yl)propylamino)phenylamino)-9H-purine, or
    8-(3-chloro-4-(3-fluorobenzyloxy)phenylamino)-9-isopropyl-2-(4-morpholinophenylamino)-9H-purine.


A second technical solution to be solved in the present invention is to provide a process for synthesizing an arylamino purine derivative represented by formula I, which process comprises:
using 2,4-dichloro-5-nitropyrimidine substituted at 6-position by a substituent group as starting material; introducing a R1-substituted amino at 4-position by a low temperature substituting process; then introducing




at 2-position by a high temperature substituting process; reducing the nitro at 5-position to amino; and finally forming a closed pyrazole ring with R3NCS (R3-substituted isothiocyanate), or





(R3-substituted methyl phenylcarbamodithioate) to obtain the targeted product.

The synthesis route is shown as below:




R1 represents —H, —C11H(2m+1), C3-C2cycloalkyl, —CmH(2m+1) substituted by C3-C2cycloalkyl, C3-C2cycloalkyl substituted by —CmH(2m+1), heterocyclyl containing 3-8 carbon atoms, amino substituted by heterocyclyl containing 3-8 carbon atoms, aryl containing 6-8 carbon atoms, or heteroaryl containing 6-8 carbon atoms; said heterocyclyl contains 1-3 heteroatoms selected from N, O and S; said heteroaryl contains 1-3 heteroatoms selected from N, O and S;

R2 represents —H, —NH2, —OH, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1), —NHCmH(2m+1), aryloxy containing 6-12 carbon atoms, or arylamino containing 6-12 carbon atoms;

R3 represents C3-C2cycloalkyl, C3-C2cycloalkyl substituted by —CmH(2m+1), aryl containing 6-80 carbon atoms, or heteroaryl containing 6-80 carbon atoms; said heteroaryl contains 1-15 heteroatoms selected from N, O and S;

R4-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —OCmH(2m+1),





m=1-8; and

n=0-4.

The advantages of said process lie in the short reaction step and the cheap and easily obtainable starting materials and agents.
A third technical solution to be solved in the present invention is to provide a use of the above-mentioned arylamino purine derivative represented by formula I in the manufacture of a medicament for the treatment of tumors.
The present invention also provides a pharmaceutical composition containing an arylamino purine derivative represented by formula I or its pharmaceutical acceptable salt. Said pharmaceutical composition can be used to prepare antitumor drugs.
The advantages of the present inventions lie in that it is proved by experiment that the present arylamino purine derivative is a multiple-kinase inhibitor that targets the kinases such as EGFR, VEGFR and MET. The present arylamino purine derivative has not only a good inhibitory effect on the non-small cell lung carcinoma with deletion mutation of exon 19 and L858R point mutation of exon 21 in EGFR, but also a good inhibitory effect on the non-small cell lung carcinoma that is resistant to Gefitinib. The present arylamino purine derivative also has a good inhibitory effect on other types of tumors. Therefore, the present arylamino purine derivative can be useful to prepare a pharmaceutical composition against these tumors. The present invention provides a new option in the field of preparing antitumor drugs, and has a good market prospect.



BRIEF DESCRIPTION OF DRAWINGS
FIG. 1 illustrates the in-vivo anti-tumor effect of Compound 8-10 and provides the tumor growth curves in a nude mouse subcutaneously transplanted human non-small cell lung carcinoma (HCC827 cell strain) model.
FIG. 2 illustrates the in-vivo anti-tumor effect of Compound 8-10 and provides a photograph of photograph of tumors obtained by dissection after the assay of the nude mouse subcutaneously transplanted human non-small cell lung carcinoma (HCC827 cell strain) model.
FIG. 3 illustrates the in-vivo anti-tumor effect of Compound 8-29 and provides the tumor growth curves in a nude mouse subcutaneously transplanted human acute myelogenous leukemia (MV4-11 cell strain) model.



DETAILED DESCRIPTION OF THE INVENTION
Hereinafter, the present invention will be further illustrated with reference to the following examples. However, these examples are only provided for illustration purposes, and are not to limit the scope of the present invention. All of the modifications made based on the above disclosures will fall into the scope of the present invention.
The reaction formulae in the Example are summarized as follows:




R1 represents —H, —CmH(2m+1), C3-C2cycloalkyl, —CmH(2m+1) substituted by C3-C2cycloalkyl, C3-C2cycloalkyl substituted by —CmH(2m+1), heterocyclyl containing 3-8 carbon atoms, amino substituted by heterocyclyl containing 3-8 carbon atoms, aryl containing 6-8 carbon atoms, or heteroaryl containing 6-8 carbon atoms; said heterocyclyl contains 1-3 heteroatoms selected from N, O and S; said heteroaryl contains 1-3 heteroatoms selected from N, O and S;

R2 represents —H, —NH2, —OH, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1), —NHCmH(2m+1), aryloxy containing 6-12 carbon atoms, or arylamino containing 6-12 carbon atoms;

R3 represents C3-C2cycloalkyl, C3-C2cycloalkyl substituted by —CmH(2m+1), aryl containing 6-80 carbon atoms, or heteroaryl containing 6-80 carbon atoms; said heteroaryl contains 1-15 heteroatoms selected from N, O and S;

R4-R8 respectively represent —H, —F, —Cl, —Br, —CF3, —OCF3, —OCmH(2m+1),





m=1-8; and

n=0-4.

Example 1
The Preparation of 2-chloro-4-amino-5-nitropyrimidine




Aqueous ammonia (8.0 ml) and N,N-diisopropylethylamine (13.2 ml) were dissolved into 150 ml dichloromethane. The mixture was added dropwise to a solution of 2,4-dichloro-5-nitropyrimidine (10.0 g) in dichloromethane (30 ml) at 0° C. After the completion of the dropwise addition, the mixture was kept at the same temperature to react for 1 hour. The precipitate was filtered off. The filter cake was recrystallized to obtain a yellow solid (8.1 g) in a yield of 90.1%. 1H NMR (400 MHz, DMSO-d6): δ 9.20 (s, 1H), 9.02 (s, 1H), 8.60 (s, 1H) ppm.
Example 2
The Preparation of 2-chloro-4-methylamino-5-nitropyrimidine




A solution of methyl amine-ethanol (7.6 ml) and N,N-diisopropylethylamine (13.2 ml) were dissolved into 150 ml dichloromethane. The mixture was added dropwise to a solution of 2,4-dichloro-5-nitropyrimidine (10.0 g) in dichloromethane (30 ml) at 0° C. After the completion of the dropwise addition, the mixture was kept at the same temperature to react for half an hour. Purification was conducted by a column chromatography to obtain a yellow solid (8.3 g) in a yield of 85.4%. 1H NMR (400 MHz, CDCl3): δ 9.05 (s, 1H), 8.41 (s, 1H), 3.22 (s, 3H) ppm.
Example 3
The Preparation of 2-chloro-4-isopropylamino-5-nitropyrimidine




Isopropylamine (4.5 ml) and N,N-diisopropylethylamine (13.2 ml) were dissolved into 150 ml dichloromethane. The mixture was added dropwise to a solution of 2,4-dichloro-5-nitropyrimidine (10.0 g) in dichloromethane (30 ml) at 0° C. After the completion of the dropwise addition, the mixture was kept at the same temperature to react for half an hour. Purification was conducted by a column chromatography to obtain a bright-yellow solid (10.1 g) in a yield of 90.4%. 1H NMR (400 MHz, CDCl3): δ 9.03 (s, 1H), 8.24 (s, 1H), 4.53 (m, 1H), 1.34 (d, J=6.8 Hz, 6H) ppm.
Example 4
The Preparation of 2-chloro-4-cyclopropylamino-5-nitropyrimidine




Cyclopropylamine (1.8 ml) and N,N-diisopropylethylamine (6.6 ml) were dissolved into 75 ml dichloromethane. The mixture was added dropwise to a solution of 2,4-dichloro-5-nitropyrimidine (5.0 g) in dichloromethane (15 ml) at 0° C. After the completion of the dropwise addition, the mixture was kept at the same temperature to react for 40 min. Purification was conducted by a column chromatography to obtain a bright-yellow solid (2.6 g) in a yield of 47%. 1H NMR (400 MHz, CDCl3): δ 8.84 (s, 1H), 7.35 (s, 1H), 3.84 (m, 1H), 1.36 (m, 4H) ppm.
Example 5
The Preparation of 2-chloro-4-cyclopentylamino-5-nitropyrimidine




Cyclopentylamine (5.17 ml) and N,N-diisopropylethylamine (12.4 ml) were dissolved into 125 ml dichloromethane. The mixture was added dropwise to a solution of 2,4-dichloro-5-nitropyrimidine (9.7 g) in dichloromethane (30 ml) at 0° C. After the completion of the dropwise addition, the mixture was kept at the same temperature to react for 80 min. Purification was conducted by a column chromatography to obtain a bright-yellow solid (7.9 g) in a yield of 65.13%. 1H NMR (400 MHz, CDCl3): δ 9.03 (s, 1H), 8.38 (s, 1H), 4.59 (m, 1H), 2.13-2.21 (m, 2H), 1.72-1.85 (m, 4H), 1.53-1.71 (m, 2H) ppm.
Example 6
The Preparation of 2-chloro-4-cyclopentylamino-5-nitro-6-methylpyrimidine




Cyclopentylamine (5.2 ml) and N,N-diisopropylethylamine (13.2 ml) were dissolved into 150 ml dichloromethane. The mixture was added dropwise to a solution of 2,4-dichloro-5-nitro-6-methylpyrimidine (10.7 g) in dichloromethane (30 ml) at 0° C. After the completion of the dropwise addition, the mixture was kept at the same temperature to react for 1 hour. Purification was conducted by a column chromatography to obtain a bright-yellow solid (11.2 g) in a yield of 84.8%. 1H NMR (400 MHz, CDCl3): δ 8.44 (s, 2H), 4.41 (m, 1H), 2.64 (s, 3H), 2.01-2.15 (m, 2H), 1.61-1.76 (m, 4H), 1.45-1.63 (m, 2H) ppm.
Example 7
The Preparation of 2-chloro-4-isopropylamino-5-nitro-6-methoxypyrimidine




Isopropylamine (4.5 ml) and N,N-diisopropylethylamine (13.2 ml) were dissolved into 150 ml dichloromethane. The mixture was added dropwise to a solution of 2,4-dichloro-5-nitro-6-methoxypyrimidine (11.5 g) in dichloromethane (30 ml) at 0° C. After the completion of the dropwise addition, the mixture was kept at the same temperature to react for 45 min. Purification was conducted by a column chromatography to obtain a yellow solid (10.9 g) in a yield of 86.1%. 1H NMR (400 MHz, CDCl3): δ 8.25 (s, 1H), 4.42 (m, 1H), 4.01 (s, 3H), 1.23 (d, J=6.8 Hz, 6H) ppm.
Example 8
The Preparation of 2-chloro-4-isopropylamino-5-nitro-6-methylaminopyrimidine




Isopropylamine (4.5 ml) and N,N-diisopropylethylamine (13.2 ml) were dissolved into 150 ml dichloromethane. The mixture was added dropwise to a solution of 2,4-dichloro-5-nitro-6-methylaminopyrimidine (11.5 g) in dichloromethane (30 ml) at 0° C. After the completion of the dropwise addition, the mixture was kept at the same temperature to react for half an hour. Purification was conducted by a column chromatography to obtain a yellow solid (10.4 g) in a yield of 82.1%. 1H NMR (400 MHz, CDCl3): δ8.17 (s, 1H), 4.48 (m, 1H), 2.78 (s, 3H), 1.31 (d, J=6.8 Hz, 6H) ppm.
Example 9
The Preparation of 2-chloro-4-isopropylamino-5-nitro-6-methylpyrimidine




Isopropylamine (4.5 ml) and N,N-diisopropylethylamine (13.2 ml) were dissolved into 150 ml dichloromethane. The mixture was added dropwise to a solution of 2,4-dichloro-5-nitro-6-methylpyrimidine (10.7 g) in dichloromethane (30 ml) at 0° C. After the completion of the dropwise addition, the mixture was kept at the same temperature to react for half an hour. Purification was conducted by a column chromatography to obtain a bright-yellow solid (10.2 g) in a yield of 86.8%. 1H NMR (400 MHz, CDCl3): δ 8.01 (s, 1H), 4.48 (m, J=4.1, 1H), 2.72 (s, 3H), 1.32 (d, J=6.8 Hz, 6H) ppm.
Example 10
The Preparation of 2-chloro-4-cyclohexylamino-5-nitropyrimidine




Cyclohexylamine (5.72 ml) and N,N-diisopropylethylamine (12.4 ml) were dissolved into 125 ml dichloromethane. The mixture was added dropwise to a solution of 2,4-dichloro-5-nitropyrimidine (9.7 g) in dichloromethane (30 ml) at 0° C. After the completion of the dropwise addition, the mixture was kept at the same temperature to react for 80 min. Purification was conducted by a column chromatography to obtain a bright-yellow solid (9.1 g) in a yield of 71.2%. ESI-MS (m/z, %) 258 (M−H)+; 1H NMR (400 MHz, CDCl3): δ9.04 (s, 1H), 8.35 (s, 1H), 4.34 (m, 1H), 2.03 (m, 2H), 1.79 (m, 2H), 1.53-1.25 (m, 6H).
Example 11
The Preparation of 2-chloro-4-cyclohexylmethylamino-5-nitropyrimidine




Cyclohexylmethylamine (3.11 g) and N,N-diisopropylethylamine (6.2 ml) were dissolved into 45 ml dichloromethane. The mixture was added dropwise to a solution of 2,4-dichloro-5-nitropyrimidine (4.85 g) in dichloromethane (30 ml) at 0° C. After the completion of the dropwise addition, the mixture was kept at the same temperature to react for 20 min. Purification was conducted by a column chromatography to obtain a bright-yellow sheet-like solid (1.73 g) in a yield of 26%. ESI-MS (m/z, %) 272 (M−H)+. 1H NMR (400 MHz, CDCl3): δ 9.03 (s, 1H), 8.48 (s, 1H), 3.52 (m, 2H), 1.78-1.64 (m, 7H), 1.33-1.14 (m, 4H), 1.08-1.00 (m, 2H).
Example 12
The Preparation of 4-(4-isopropylamino-5-nitropyrimidin-2-ylamino)-N-(4-methylpiperazin-1-yl)benzamide




4-amino-N-(4-methylpiperazin-1-yl)benzamide (4.7 g) was added to a solution of Compound 2-3 (4.3 g) in n-butanol (150 ml). The mixture was reacted at 90° C. for 3.5 hours, cooled to room temperature, filtered, washed and dried to obtain a yellow solid (5.9 g) in a yield of 71.2%. 1H NMR (400 MHz, DMSO-d6): δ 10.01 (s, 1H), 9.32 (s, 1H), 8.78 (s, 1H), 8.21 (m, 2H), 7.84 (s, 1H), 7.63 (m, 2H), 4.31 (m, 1H), 2.75 (t, J=4.8 Hz, 4H), 2.38 (br, 4H), 2.13 (s, 3H), 1.19 (d, J=6.8 Hz, 6H) ppm.
Example 13
The Preparation of 3-fluoro-4-(4-isopropylamino-5-nitropyrimidin-2-ylamino)-N-(4-methylpiperazin-1-yl)benzamide




4-amino-3-fluoro-N-(4-methylpiperazin-1-yl)benzamide (5.0 g) was added to a solution of Compound 2-3 (4.3 g) in n-butanol (150 ml). The mixture was reacted at 90° C. for 4 hours, cooled to room temperature, filtered, washed and dried to obtain a yellow solid (6.5 g) in a yield of 75.7%. 1H NMR (400 MHz, DMSO-d6): δ 10.11 (s, 1H), 9.47 (s, 1H), 8.99 (s, 1H), 8.38 (d, J=7.6 Hz, 1H), 7.86 (m, 1H), 7.65 (m, 2H), 4.27 (m, 1H), 2.89 (t, J=4.8 Hz, 4H), 2.42 (br, 4H), 2.19 (s, 3H), 1.23 (d, J=6.4 Hz, 6H) ppm.
Example 14
The Preparation of 3-methoxy-4-(4-isopropylamino-5-nitropyrimidin-2-ylamino)-N-(4-methylpiperazin-1-yl)benzamide




4-amino-3-methoxy-N-(4-methylpiperazin-1-yl)benzamide (5.3 g) was added to a solution of Compound 2-3 (4.3 g) in n-butanol (150 ml). The mixture was reacted at 90° C. for 4.5 hours, cooled to room temperature, filtered, washed and dried to obtain a yellow solid (6.8 g) in a yield of 77.1%. 1H NMR (400 MHz, DMSO-d6): δ 9.13 (s, 1H), 8.38 (d, J=6.8 Hz, 1H), 7.93 (m, 1H), 7.63 (d, J=8.0 Hz, 1H), 7.46 (m, 1H), 7.08 (m, 1H), 7.01 (m, 1H), 4.31 (m, 1H), 3.86 (s, 3H), 3.55 (br, 4H), 2.52 (br, 4H), 2.32 (s, 3H), 1.24 (d, J=6.4 Hz, 6H) ppm.
Example 15
The Preparation of 4-(4-isopropylamino-5-nitropyrimidin-2-ylamino)-N-(1-methylpiperidin-4-yl)benzamide




4-amino-N-(4-methylpiperidin-1-yl)benzamide (4.6 g) was added to a solution of Compound 2-3 (4.3 g) in n-butanol (150 ml). The mixture was reacted at 90° C. for 4.5 hours, cooled to room temperature, filtered, washed and dried to obtain a yellow solid (5.7 g) in a yield of 70.0%. 1H NMR (400 MHz, DMSO-d6): δ 10.58 (s, 1H), 9.02 (s, 1H), 8.48 (d, J=5.6 Hz, 1H), 8.20 (d, J=6.8 Hz, 1H), 7.86 (m, 4H), 4.45 (m, 1H), 3.80 (m, 1H), 2.94 (br, 4H), 2.32 (s, 3H), 1.62-1.83 (m, 4H), 1.33 (d, J=6.4 Hz, 6H) ppm.
Example 16
The Preparation of 4-amino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-5-nitropyrimidine




4-(4-methylpiperazinyl)phenylamine (3.8 g) was added to a solution of Compound 2-1 (3.5 g) in n-butanol (150 ml). The mixture was reacted at 90° C. for 4.5 hours, cooled to room temperature, filtered, washed and dried to obtain a red solid (5.2 g) in a yield of 79.5%. 1H NMR (400 MHz, CDCl3): δ 9.07 (s, 1H), 8.52 (s, 2H), 8.40 (s, 1H), 7.57 (s, 1H), 7.51 (s, 1H), 7.10 (m, 2H), 3.31 (t, J=4.8 Hz, 4H), 2.81 (t, J=4.8 Hz, 4H), 2.30 (s, 3H) ppm.
Example 17
The Preparation of 4-methylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-5-nitropyrimidine




4-(4-methylpiperazinyl)phenylamine (3.8 g) was added to a solution of Compound 2-2 (3.8 g) in n-butanol (150 ml). The mixture was reacted at 90° C. for 3.5 hours, cooled to room temperature, filtered, washed and dried to obtain a red solid (5.1 g) in a yield of 74.8%. 1H NMR (400 MHz, CDCl3): δ 9.11 (s, 1H), 8.34 (s, 1H), 7.59 (s, 1H), 7.51 (m, 2H), 7.23 (m, 2H), 4.21 (s, 3H), 3.15 (t, J=4.8 Hz, 4H), 2.87 (t, J=4.8 Hz, 4H), 2.48 (s, 3H) ppm.
Example 18
The Preparation of 4-isopropylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-5-nitropyrimidine




4-(4-methylpiperazinyl)phenylamine (3.8 g) was added to a solution of Compound 2-3 (4.3 g) in n-butanol (150 ml). The mixture was reacted at 90° C. for 3 hours, cooled to room temperature, filtered, washed and dried to obtain a red solid (6.2 g) in a yield of 84.1%. 1H NMR (400 MHz, CDCl3): δ 9.02 (s, 1H), 8.42 (s, 1H), 7.63 (s, 1H), 7.51 (s, 2H), 6.95 (m, 2H), 4.41 (m, 1H), 3.22 (t, J=4.8 Hz, 4H), 2.61 (t, J=4.8 Hz, 4H), 2.37 (s, 3H), 1.33 (d, J=6.4 Hz, 6H) ppm.
Example 19
The Preparation of 4-cyclopropylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-5-nitropyrimidine




4-(4-methylpiperazinyl)phenylamine (3.8 g) was added to a solution of Compound 2-4 (4.3 g) in n-butanol (150 ml). The mixture was reacted at 90° C. for 3.5 hours, cooled to room temperature, filtered, washed and dried to obtain a red solid (6.4 g) in a yield of 87.2%. 1H NMR (400 MHz, CDCl3): δ 9.08 (s, 1H), 8.39 (s, 1H), 7.90 (s, 1H), 7.58 (m, 2H), 6.94 (m, 2H), 4.32 (m, 1H), 3.05 (t, J=4.8 Hz, 4H), 2.90 (t, J=4.8 Hz, 4H), 2.67 (s, 3H), 1.39 (m, 4H) ppm.
Example 20
The Preparation of 4-cyclopentylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-5-nitropyrimidine




4-(4-methylpiperazinyl)phenylamine (3.8 g) was added to a solution of Compound 2-5 (4.8 g) in n-butanol (150 ml). The mixture was reacted at 90° C. for 3 hours, cooled to room temperature, filtered, washed and dried to obtain a red solid (6.0 g) in a yield of 76.0%. 1H NMR (400 MHz, CDCl3): δ 9.03 (s, 1H), 8.47 (s, 1H), 7.69 (s, 1H), 7.51 (m, 2H), 7.11 (m, 2H), 4.43 (m, 1H), 3.28 (t, J=4.8 Hz, 4H), 2.67 (t, J=4.8 Hz, 4H), 2.45 (s, 3H), 2.15-2.23 (m, 2H), 1.74-1.86 (m, 4H), 1.50-1.72 (m, 2H) ppm.
Example 21
The Preparation of 4-cyclopentylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-5-nitro-6-methylpyrimidine




4-(4-methylpiperazinyl)phenylamine (3.8 g) was added to a solution of Compound 2-6 (5.1 g) in n-butanol (150 ml). The mixture was reacted at 90° C. for 5 hours, cooled to room temperature, filtered, washed and dried to obtain a red solid (6.4 g) in a yield of 78.3%. 1H NMR (400 MHz, CDCl3): δ 8.42 (s, 1H), 7.62 (s, 1H), 7.41 (s, 2H), 6.98 (m, 2H), 4.21 (m, 1H), 3.14 (t, J=4.8 Hz, 4H), 2.56 (t, J=4.8 Hz, 4H), 2.51 (s, 3H), 2.34 (s, 3H), 1.47-1.85 (m, 8H) ppm.
Example 22
The Preparation of 4-isopropylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-5-nitro-6-methoxypyrimidine




4-(4-methylpiperazinyl)phenylamine (3.8 g) was added to a solution of Compound 2-7 (4.9 g) in n-butanol (150 ml). The mixture was reacted at 90° C. for 5.5 hours, cooled to room temperature, filtered, washed and dried to obtain a reddish-brown solid (6.5 g) in a yield of 81.5%. 1H NMR (400 MHz, CDCl3): δ 8.51 (s, 1H), 7.86 (s, 1H), 7.43 (s, 2H), 7.01 (m, 2H), 4.32 (m, 1H), 3.94 (s, 3H), 3.08 (t, J=4.8 Hz, 4H), 2.64 (t, J=4.8 Hz, 4H), 2.53 (s, 3H), 1.43 (d, J=6.4 Hz, 6H) ppm.
Example 23
The Preparation of 4-isopropylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-5-nitro-6-methylaminopyrimidine




4-(4-methylpiperazinyl)phenylamine (3.8 g) was added to a solution of Compound 2-8 (4.9 g) in n-butanol (150 ml). The mixture was reacted at 90° C. for 6 hours, cooled to room temperature, filtered, washed and dried to obtain a reddish-brown solid (6.0 g) in a yield of 75.4%. 1H NMR (400 MHz, CDCl3): δ 8.58 (s, 1H), 7.81 (s, 1H), 7.47 (s, 2H), 7.08 (m, 2H), 4.45 (m, 1H), 3.14 (t, J=4.8 Hz, 4H), 2.79 (s, 3H), 2.54 (t, J=4.8 Hz, 4H), 2.42 (s, 3H), 1.45 (d, J=6.4 Hz, 6H) ppm.
Example 24
The Preparation of 4-isopropylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-5-nitro-6-methylpyrimidine




4-(4-methylpiperazinyl)phenylamine (3.8 g) was added to a solution of Compound 2-9 (4.6 g) in n-butanol (150 ml). The mixture was reacted at 90° C. for 3.5 hours, cooled to room temperature, filtered, washed and dried to obtain a red solid (6.3 g) in a yield of 82.3%. 1H NMR (400 MHz, DMSO-d6): δ 11.01 (s, 1H), 10.15 (s, 1H), 7.66 (d, J=8.4, 2H), 6.99 (d, J=9.2, 2H), 4.35 (s, 1H), 3.76 (d, J=11.2, 2H), 3.46 (d, J=10.8, 2H), 3.11 (m, J=13.6, 4H), 2.79 (s, 3H), 2.60 (s, 3H), 1.27 (d, J=6.4, 6H) ppm.
Example 25
The Preparation of 4-(4-cyclopentylamino-5-nitropyrimidin-2-ylamino)-N-(4-methylpiperazin-1-yl)benzamide




4-amino-N-(4-methylpiperazin-1-yl)benzamide (4.85 g) was added to a solution of Compound 2-5 (4.7 g) in n-butanol (150 ml). The mixture was reacted at 90° C. for 4.5 hours, cooled to room temperature, filtered, washed and dried to obtain a yellow solid (6.4 g) in a yield of 72.6%. MS m/z (ESI): 441 [M+H]+.
Example 26
The Preparation of 4-(4-isopropylamino-5-nitropyrimidin-2-ylamino)-N-(1-methylpiperidin-4-yl)benzamide




4-amino-3-fluoro-N-(4-methylpiperidin-1-yl)benzamide (5.0 g) was added to a solution of Compound 2-3 (4.3 g) in n-butanol (150 ml). The mixture was reacted at 90° C. for 4.0 hours, cooled to room temperature, filtered, washed and dried to obtain a yellow solid (6.3 g) in a yield of 72.8%. MS m/z (ESI): 432 [M+H]+.
Example 27
The Preparation of N-(3-(4-isopropylamino-5-nitropyrimidin-2-ylamino)phenyl)acrylamide




N-(3-aminophenyl)acrylamide (3.2 g) was added to a solution of Compound 2-3 (4.3 g) in n-butanol (150 ml). The mixture was reacted at 90° C. for 3 hours, cooled to room temperature, filtered, washed and dried to obtain a red solid (6.0 g) in a yield of 88.3%. MS m/z (ESI): 344 [M+H]+.
Example 28
The Preparation of 4-isopropylamino-2-(4-(2-morpholinoethoxy)phenylamino)-5-nitropyrimidine




4-(2-morpholinoethoxy)phenylamine (4.4 g) was added to a solution of Compound 2-3 (4.3 g) in n-butanol (150 ml). The mixture was reacted at 90° C. for 3.5 hours, cooled to room temperature, filtered, washed and dried to obtain a red solid (6.4 g) in a yield of 80.0%. MS m/z (ESI): 403 [M+H]+.
Example 29
The Preparation of 4-isopropylamino-2-(4-(3-(4-methylpiperazin-1-yl)propylamino)phenylamino)-5-nitropyrimidine




4-(3-(4-methylpiperazin-1-yl)propylamino)phenylamine (4.9 g) was added to a solution of Compound 2-3 (4.3 g) in n-butanol (150 ml). The mixture was reacted at 90° C. for 4 hours, cooled to room temperature, filtered, washed and dried to obtain a red solid (6.8 g) in a yield of 79.9%. MS m/z (ESI): 429 [M+H]+.
Example 30
The Preparation of 4-isopropylamino-2-(4-morpholinophenylamino)-5-nitropyrimidine




4-morpholinophenylamine (3.6 g) was added to a solution of Compound 2-3 (4.3 g) in n-butanol (150 ml). The mixture was reacted at 90° C. for 4 hours, cooled to room temperature, filtered, washed and dried to obtain a red solid (5.3 g) in a yield of 74.7%. MS m/z (ESI): 359 [M+H]+.
Example 31
The Preparation of 4-cyclopentylamino-2-(4-(4-morpholinylmethyl)phenylamino)-5-nitropyrimidine




4-morpholinomethylphenylamine (3.8 g) was added to a solution of Compound 2-5 (4.8 g) in n-butanol (150 ml). The mixture was reacted at 90° C. for 4.0 hours, cooled to room temperature, filtered, washed and dried to obtain a yellow solid (6.3 g) in a yield of 80.6%. MS m/z (ESI): 399 [M+H]+.
Example 32
The Preparation of 4-cyclohexylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-5-nitropyrimidine




4-(4-methylpiperazinyl)phenylamine (3.1 g) was added to a solution of Compound 2-10 (2.3 g) in n-butanol (40 ml). The mixture was reacted at 90° C. for 4.0 hours, cooled to room temperature, filtered, washed and dried to obtain a red solid (4.13 g) in a yield of 83.8%. MS m/z (ESI): 412 [M+H]+. 1H NMR (400 MHz, DMSO-d6): δ 10.71 (s, 1H), 10.35 (s, 1H), 8.96 (s, 1H), 8.48 (d, J=6.8 Hz, 1H), 7.71 (d, J=8.8 Hz, 1H), 6.99 (d, J=9.2 Hz, 2H), 4.04 (m, 1H), 3.78 (m, 2H), 3.46 (m, 2H), 3.15 (m, 2H), 3.04 (m, 2H), 2.83 (s, 1H), 1.98 (m, 2H), 1.65 (m, 1H), 1.43 (m, 4H), 1.26 (m, 1H) ppm.
Example 33
The Preparation of 4-cyclopentylamino-2-(4-(2-methoxyethoxy)phenylamino)-5-nitropyrimidine




4-(2-methoxyethoxy)phenylamine (2.7 g) was added to a solution of Compound 2-5 (3.7 g) in n-butanol (80 ml). The mixture was reacted at 90° C. for 4.0 hours, cooled to room temperature, filtered, washed and dried to obtain a yellow floc-like solid (4.53 g) in a yield of 80.9%. MS m/z (ESI): 374 [M+H]+; 1H NMR (400 MHz, DMSO-d6): δ10.34 (s, 1H), 8.95 (s, 1H), 8.50 (s, 1H), 7.70 (d, J=6.0 Hz, 2H), 6.93 (d, J=8.8 Hz, 2H), 4.43 (m, 1H), 4.07 (m, 2H), 3.64 (m, 2H), 3.30 (s, 3H), 2.03 (m, 2H), 1.72 (m, 2H), 1.61 (m, 4H) ppm.
Example 34
The Preparation of 4-cyclopentylamino-2-(4-((4-ethyl piperazin-1-yl)methyl)phenylamino)-5-nitropyrimidine




4-((4-ethylpiperazin-1-yl)methyl)phenylamine (3.6 g) was added to a solution of Compound 2-5 (6.0 g) in n-butanol (130 ml). The mixture was reacted at 90° C. for 4.0 hours, cooled to room temperature, filtered, washed and dried to obtain a red solid in a yield of 81.8%. MS m/z (ESI): 426 [M+H]+.
Example 35
The Preparation of 4-cyclopentylamino-2-(4-((4-methylpiperazin-1-yl)methyl)phenylamino)-5-nitropyrimidine




4-((4-methylpiperazin-1-yl)methyl)phenylamine (4.7 g) was added to a solution of Compound 2-5 (5.28 g) in n-butanol (130 ml). The mixture was reacted at 90° C. for 4.0 hours, cooled to room temperature, filtered, washed and dried to obtain a red solid in a yield of 85.7%. MS m/z (ESI): 412 [M+H]+.
Example 36
The Preparation of 4-cyclohexylmethylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-5-nitropyrimidine




4-(4-methylpiperazinyl)phenylamine (1.13 g) was added to a solution of Compound 2-11 (1.6 g) in n-butanol (25 ml). The mixture was reacted at 90° C. for 4.0 hours, cooled to room temperature, filtered, washed and dried to obtain an orange-red solid (4.13 g) in a yield of 87.6%. MS m/z (ESI): 426 [M+H]+.
Example 37
The Preparation of 4-(4-isopropylamino-5-aminopyrimidin-2-ylamino)-N-(4-methylpiperazin-1-yl)benzamide




A solution of rongalite (sodium dithionite) (9.6 g) in water (30 ml) was added to a solution of 4-1 (2.1 g) in tetrahydrofuran (25 ml). The mixture was stirred at room temperature for 6-12 hours, adjusted to a pH of 7-8 by adding a saturated potassium carbonate solution, and then extracted with dichloromethane (5×20 ml). The organic phase was dried over anhydrous sodium sulfate, and rotary evaporated to dryness to obtain a jade-green solid (1.2 g) in a yield of 63.2%. 1H NMR (400 MHz, DMSO-d6): δ 10.01 (s, 1H), 9.32 (s, 1H), 8.78 (s, 1H), 8.01 (m, 2H), 7.83 (s, 1H), 7.72 (m, 2H), 4.29 (m, 1H), 4.08 (s, 2H), 2.87 (t, J=4.8 Hz, 4H), 2.47 (br, 4H), 2.24 (s, 3H), 1.22 (d, J=6.8 Hz, 6H) ppm.
Example 38
The Preparation of 3-fluoro-4-(4-isopropylamino-5-aminopyrimidin-2-ylamino)-N-(4-methylpiperazin-1-yl)benzamide




A solution of rongalite (9.6 g) in water (30 ml) was added to a solution of 4-2 (2.2 g) in tetrahydrofuran (25 ml). The mixture was stirred at room temperature for 6-12 hours, adjusted to a pH of 7-8 by adding a saturated potassium carbonate solution, and then extracted with dichloromethane (5×20 ml). The organic phase was dried over anhydrous sodium sulfate, and rotary evaporated to dryness to obtain a jade-green solid (1.4 g) in a yield of 68.4%. 1H NMR (400 MHz, DMSO-d6): δ9.26 (s, 1H), 8.44 (m, 1H), 7.81 (s, 1H), 7.56 (d, J=10.8 Hz, 2H), 7.40 (s, 1H), 6.24 (d, J=7.2 Hz, 1H), 4.31 (s, 2H), 4.22 (m, 1H), 2.86 (t, J=4.8 Hz, 4H), 2.41 (br, 4H), 2.18 (s, 3H), 1.22 (d, J=6.4 Hz, 6H) ppm.
Example 39
The Preparation of 3-methoxy-4-(4-isopropylamino-5-aminopyrimidin-2-ylamino)-N-(4-methylpiperazin-1-yl)benzamide




In a 500 ml round-bottom flask, Compound 4-3 (4.0 g) was dissolved into 150 ml ethanol, and then 40 ml water and 5.0 g NH4Cl were added thereto. The mixture was stirred at room temperature for 10 minutes, and warmed to 90° C. 2.7 g Fe powder was added to the mixture in three portions. TLC detection indicated the completion of the reaction of the starting material. The reaction time was 3.5 hours. The mixture was filtered while hot, rotary evaporated to dryness, and subjected to a column chromatography to obtain a bluish-black solid (2.8 g) in a yield of 75.0%. MS m/z (ESI): 415 [M+H]+.
Example 40
The Preparation of 4-(4-isopropylamino-5-aminopyrimidin-2-ylamino)-N-(1-methylpiperidin-4-yl)benzamide




In a 500 ml round-bottom flask, Compound 4-4 (4.0 g) was dissolved into 150 ml ethanol, and then 40 ml water and 5.0 g NH4Cl were added thereto. The mixture was stirred at room temperature for 10 minutes, and warmed to 90° C. 2.7 g Fe powder was added to the mixture in three portions. TLC detection indicated the completion of the reaction of the starting material. The reaction time was 4.2 hours. The mixture was filtered while hot, rotary evaporated to dryness, and subjected to a column chromatography to obtain a bluish-black solid (2.6 g) in a yield of 70.8%. MS m/z (ESI): 384 [M+H]+.
Example 41
The Preparation of 4-amino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-5-aminopyrimidine




In a 500 ml round-bottom flask, Compound 4-5 (6.6 g) was dissolved into 160 ml ethanol, and then 40 ml water and 5.35 g NH4Cl were added thereto. The mixture was stirred at room temperature for 10 minutes, and warmed to 90° C. 6.6 g Fe powder was added to the mixture in three portions. TLC detection indicated the completion of the reaction of the starting material. The reaction time was 4.5 hours. The mixture was filtered while hot, rotary evaporated to dryness, and recrystallized to obtain a bluish-black solid in a yield of 71.0%. 1H NMR (400 MHz, DMSO-d6): δ 8.18 (s, 1H), 7.56 (d, J=9.2 Hz, 2H), 6.79 (d, J=8.8 Hz, 2H), 6.00 (d, J=7.6 Hz, 1H), 4.24 (m, 1H), 4.01 (s, 2H), 2.99 (t, J=4.4 Hz, 4H), 2.43 (t, J=4.8 Hz, 4H), 2.21 (s, 3H), 1.21 (d, J=6.4 Hz, 6H) ppm.
Example 42
The Preparation of 4-methylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-5-aminopyrimidine




In a 100 ml round-bottom flask, Compound 4-6 (1.0 g) was dissolved into 60 ml ethanol, and then 15 ml water and 0.8 g NH4Cl were added thereto. The mixture was stirred at room temperature for 10 minutes, and warmed to 90° C. 0.84 g Fe powder was added to the mixture in three portions. TLC detection indicated the completion of the reaction of the starting material. The reaction time was 1.5 hours. The mixture was filtered while hot, rotary evaporated to dryness, and subjected to a column chromatography to obtain a bluish-black solid (0.7 g) in a yield of 70.2%. MS m/z (ESI): 314 [M+H]+.
Example 43
The Preparation of 4-isopropylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-5-aminopyrimidine




In a 500 ml round-bottom flask, Compound 4-7 (3.7 g) was dissolved into 200 ml ethanol, and then 50 ml water and 2.7 g NH4Cl were added thereto. The mixture was stirred at room temperature for 10 minutes, and warmed to 90° C. 2.8 g Fe powder was added to the mixture in three portions. TLC detection indicated the completion of the reaction of the starting material. The reaction time was 3 hours. The mixture was filtered while hot, rotary evaporated to dryness, and subjected to a column chromatography to obtain a bluish-black solid (2.4 g) in a yield of 70.6%. 1H NMR (400 MHz, DMSO-d6): δ 8.18 (s, 1H), 7.56 (d, J=9.2 Hz, 2H), 6.79 (d, J=8.8 Hz, 2H), 6.00 (d, J=7.6 Hz, 1H), 4.24 (m, 1H), 4.01 (s, 2H), 2.99 (t, J=4.4 Hz, 4H), 2.43 (t, J=4.8 Hz, 4H), 2.21 (s, 3H), 1.21 (d, J=6.4 Hz, 6H) ppm.
Example 44
The Preparation of 4-cyclopropylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-5-aminopyrimidine




In a 100 ml round-bottom flask, Compound 4-8 (1.1 g) was dissolved into 60 ml ethanol, and then 15 ml water and 0.8 g NH4Cl were added thereto. The mixture was stirred at room temperature for 10 minutes, and warmed to 90° C. 0.84 g Fe powder was added to the mixture in three portions. TLC detection indicated the completion of the reaction of the starting material. The reaction time was 1.5 hours. The mixture was filtered while hot, rotary evaporated to dryness, and subjected to a column chromatography to obtain a column chromatography to obtain bluish-black solid (0.8 g) in a yield of 79.2%. MS m/z (ESI): 340 [M+H]+.
Example 45
The Preparation of 4-cyclopentylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-5-aminopyrimidine




In a 250 ml round-bottom flask, Compound 4-9 (2.0 g) was dissolved into 100 ml ethanol, and then 25 ml water and 1.4 g NH4Cl were added thereto. The mixture was stirred at room temperature for 10 minutes, and warmed to 90° C. 1.4 g Fe powder was added to the mixture in three portions. TLC detection indicated the completion of the reaction of the starting material. The reaction time was 2.5 hours. The mixture was filtered while hot, rotary evaporated to dryness, and recrystallized to obtain a bluish-black solid (1.1 g) in a yield of 59.3%. MS m/z (ESI): 368 [M+H]+.
Example 46
The Preparation of 4-cyclopentylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-5-amino-6-methylpyrimidine




In a 500 ml round-bottom flask, Compound 4-10 (3.7 g) was dissolved into 200 ml ethanol, and then 50 ml water and 2.7 g NH4Cl were added thereto. The mixture was stirred at room temperature for 10 minutes, and warmed to 90° C. 2.8 g Fe powder was added to the mixture in three portions. TLC detection indicated the completion of the reaction of the starting material. The reaction time was 3 hours. The mixture was filtered while hot, rotary evaporated to dryness, and subjected to a column chromatography to obtain a bluish-black solid (2.6 g) in a yield of 75.7%. MS m/z (ESI): 382 [M+H]+.
Example 47
The Preparation of 4-isopropylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-5-amino-6-methoxypyrimidine




In a 500 ml round-bottom flask, Compound 4-11 (3.9 g) was dissolved into 200 ml ethanol, and then 50 ml water and 2.7 g NH4Cl were added thereto. The mixture was stirred at room temperature for 10 minutes, and warmed to 90° C. 2.8 g Fe powder was added to the mixture in three portions. TLC detection indicated the completion of the reaction of the starting material. The reaction time was 3 hours. The mixture was filtered while hot, rotary evaporated to dryness, and subjected to a column chromatography to obtain a bluish-black solid (2.5 g) in a yield of 70.0%. MS m/z (ESI): 372 [M+H]+.
Example 48
The Preparation of 4-isopropylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-5-amino-6-methylaminopyrimidine




In a 500 ml round-bottom flask, Compound 4-12 (3.8 g) was dissolved into 200 ml ethanol, and then 50 ml water and 2.7 g NH4Cl were added thereto. The mixture was stirred at room temperature for 10 minutes, and warmed to 90° C. 2.8 g Fe powder was added to the mixture in three portions. TLC detection indicated the completion of the reaction of the starting material. The reaction time was 3 hours. The mixture was filtered while hot, rotary evaporated to dryness, and subjected to a column chromatography to obtain a bluish-black solid (2.4 g) in a yield of 67.3%. MS m/z (ESI): 371 [M+H]+.
Example 49
The Preparation of 4-isopropylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-5-amino-6-methylpyrimidine




In a 500 ml round-bottom flask, Compound 4-13 (3.7 g) was dissolved into 200 ml ethanol, and then 50 ml water and 2.7 g NH4Cl were added thereto. The mixture was stirred at room temperature for 10 minutes, and warmed to 90° C. 2.8 g Fe powder was added to the mixture in three portions. TLC detection indicated the completion of the reaction of the starting material. The reaction time was 3 hours. The mixture was filtered while hot, rotary evaporated to dryness, and subjected to a column chromatography to obtain a bluish-black solid (2.5 g) in a yield of 73.3%. 1H NMR (400 MHz, CDCl3): δ 7.51 (d, J=8.8, 2H), 6.89 (d, J=8.8, 2H), 6.59 (s, 1H), 5.17 (d, J=7.2, 1H), 4.63 (s, 2H), 4.22 (m, J=6.72, 1H), 3.14 (t, J=5.0, 4H), 2.59 (t, J=4.8, 4H), 2.35 (s, 3H), 2.25 (s, 3H), 1.26 (d, J=6.4, 6H) ppm.
Example 50
The Preparation of 4-(4-cyclopentylamino-5-aminopyrimidin-2-ylamino)-N-(4-methylpiperazin-1-yl)benzamide




In a 500 ml round-bottom flask, Compound 4-14 (4.4 g) was dissolved into 150 ml ethanol, and then 40 ml water and 5.0 g NH4Cl were added thereto. The mixture was stirred at room temperature for 10 minutes, and warmed to 90° C. 2.7 g Fe powder was added to the mixture in three portions. TLC detection indicated the completion of the reaction of the starting material. The reaction time was 3.5 hours. The mixture was filtered while hot, rotary evaporated to dryness, and recrystallized to obtain a bluish-black solid (3.2 g) in a yield of 77.9%. MS m/z (ESI): 411 [M+H]+.
Example 51
The Preparation of 4-(4-isopropylamino-5-aminopyrimidin-2-ylamino)-N-(1-methylpiperidin-4-yl)benzamide




A solution of rongalite (9.6 g) in water (30 ml) was added to a solution of 4-15 (2.2 g) in tetrahydrofuran (25 ml). The mixture was stirred at room temperature for 6-12 hours, adjusted to a pH of 7-8 by adding a saturated potassium carbonate solution, and then extracted with dichloromethane (5×20 ml). The organic phase was dried over anhydrous sodium sulfate, and rotary evaporated to dryness to obtain a jade-green solid (1.5 g) in a yield of 69.6%. MS m/z (ESI): 402 [M+H]+.
Example 52
The Preparation of N-(3-(4-isopropylamino-5-nitropyrimidin-2-ylamino)phenyl)acrylamide




N-(3-aminophenyl)acrylamide (3.2 g) was added to a solution of Compound 2-3 (4.3 g) in n-butanol (150 ml). The mixture was reacted at 90° C. for 3 hours, cooled to room temperature, filtered, washed and dried to obtain a red solid (6.0 g) in a yield of 88.3%. MS m/z (ESI): 344 [M+H]+.
Example 53
The Preparation of 4-isopropylamino-2-(4-(2-morpholinoethoxy)phenylamino)-5-aminopyrimidine




In a 100 ml round-bottom flask, Compound 4-17 (1.21 g) was dissolved into 60 ml ethanol, and then 15 ml water and 0.8 g NH4Cl were added thereto. The mixture was stirred at room temperature for 10 minutes, and warmed to 90° C. 0.84 g Fe powder was added to the mixture in three portions. TLC detection indicated the completion of the reaction of the starting material. The reaction time was 1.5 hours. The mixture was filtered while hot, rotary evaporated to dryness, and subjected to a column chromatography to obtain a bluish-black solid in a yield of 75.2%. MS m/z (ESI): 373 [M+H]+.
Example 54
The Preparation of 4-isopropylamino-2-(4-(3-(4-methylpiperazin-1-yl)propylamino)phenylamino)-5-aminopyrimidine




In a 100 ml round-bottom flask, Compound 4-18 (1.3 g) was dissolved into 60 ml ethanol, and then 15 ml water and 0.8 g NH4Cl were added thereto. The mixture was stirred at room temperature for 10 minutes, and warmed to 90° C. 0.84 g Fe powder was added to the mixture in three portions. TLC detection indicated the completion of the reaction of the starting material. The reaction time was 1.5 hours. The mixture was filtered while hot, rotary evaporated to dryness, and subjected to a column chromatography to obtain a bluish-black solid in a yield of 78.9%. MS m/z (ESI): 399 [M+H]+.
Example 55
The Preparation of 4-isopropylamino-2-(4-morpholinophenylamino)-5-aminopyrimidine




In a 100 ml round-bottom flask, Compound 4-19 (1.1 g) was dissolved into 60 ml ethanol, and then 15 ml water and 0.8 g NH4Cl were added thereto. The mixture was stirred at room temperature for 10 minutes, and warmed to 90° C. 0.84 g Fe powder was added to the mixture in three portions. TLC detection indicated the completion of the reaction of the starting material. The reaction time was 2.5 hours. The mixture was filtered while hot, rotary evaporated to dryness, and subjected to a column chromatography to obtain a bluish-black solid in a yield of 74.1%. MS m/z (ESI): 329 [M+H]+.
Example 56
The Preparation of 4-cyclopentylamino-2-(4-(4-morpholinylmethyl)phenylamino)-5-aminopyrimidine




In a 500 ml round-bottom flask, Compound 4-20 (3.85 g) was dissolved into 150 ml ethanol, and then 40 ml water and 5.0 g NH4Cl were added thereto. The mixture was stirred at room temperature for 10 minutes, and warmed to 90° C. 2.7 g Fe powder was added to the mixture in three portions. TLC detection indicated the completion of the reaction of the starting material. The reaction time was 3.5 hours. The mixture was filtered while hot, rotary evaporated to dryness, and subjected to a column chromatography to obtain a bluish-black solid in a yield of 74.5%. MS m/z (ESI): 369 [M+H]+.
Example 57
The Preparation of 4-cyclohexylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-5-aminopyrimidine




In a 100 ml round-bottom flask, Compound 4-21 (1.03 g) was dissolved into 40 ml ethanol, and then 10 ml water and 0.67 g NH4Cl were added thereto. The mixture was stirred at room temperature for 10 minutes, and warmed to 90° C. 0.7 g Fe powder was added to the mixture in three portions. TLC detection indicated the completion of the reaction of the starting material. The reaction time was 3.7 hours. The mixture was filtered while hot, and rotary-evaporated to dryness to obtain a crude product in a yield of 78.9%. MS m/z (ESI): 382 [M+H]+.
Example 58
The Preparation of 4-cyclopentylamino-2-(4-(2-methoxyethoxy)phenylamino)-5-aminopyrimidine




In a 250 ml round-bottom flask, Compound 4-22 (3.73 g) was dissolved into 120 ml ethanol, and then 30 ml water and 2.67 g NH4Cl were added thereto. The mixture was stirred at room temperature for 10 minutes, and warmed to 90° C. 2.8 g Fe powder was added to the mixture in three portions. TLC detection indicated the completion of the reaction of the starting material. The reaction time was 4 hours. The mixture was filtered while hot, and rotary-evaporated to dryness to obtain a crude product in a yield of 84.6%. MS m/z (ESI): 344 [M+H]+.
Example 59
The Preparation of 4-cyclopentylamino-2-(4-((4-ethyl piperazin-1-yl)methyl)phenylamino)-5-aminopyrimidine




In a 500 ml round-bottom flask, Compound 4-23 (10 g) was dissolved into 240 ml ethanol, and then 60 ml water and 2.67 g NH4Cl were added thereto. The mixture was stirred at room temperature for 10 minutes, and warmed to 90° C. 6.6 g Fe powder was added to the mixture in three portions. TLC detection indicated the completion of the reaction of the starting material. The reaction time was 4 hours. The mixture was filtered while hot, and rotary-evaporated to dryness to obtain a crude product in a yield of 82.7%. MS m/z (ESI): 396 [M+H]+.
Example 60
The Preparation of 4-cyclopentylamino-2-(4-((4-methyl piperazin-1-yl)methyl)phenylamino)-5-aminopyrimidine




In a 500 ml round-bottom flask, Compound 4-24 (9.42 g) was dissolved into 240 ml ethanol, and then 60 ml water and 1.92 g NH4Cl were added thereto. The mixture was stirred at room temperature for 10 minutes, and warmed to 90° C. 6.85 g Fe powder was added to the mixture in three portions. TLC detection indicated the completion of the reaction of the starting material. The reaction time was 4 hours. The mixture was filtered while hot, and rotary-evaporated to dryness to obtain a crude product in a yield of 80.6%. MS m/z (ESI): 382 [M+H]+.
Example 61
The Preparation of 4-cyclohexylmethylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-5-aminopyrimidine




In a 100 ml round-bottom flask, Compound 4-23 (2.0 g) was dissolved into 30 ml ethanol, and then 10 ml water and 0.62 g NH4Cl were added thereto. The mixture was stirred at room temperature for 10 minutes, and warmed to 90° C. 1.29 g Fe powder was added to the mixture in three portions. TLC detection indicated the completion of the reaction of the starting material. The mixture was filtered while hot, and rotary-evaporated to dryness to obtain a crude product in a yield of 81.4%. MS m/z (ESI): 396 [M+H]+.
Example 62
The Preparation of 4-(9-isopropyl-8-phenylamino-9H-purin-2-ylamino)-N-(4-methylpiperidin-1-yl)benzamide




Compound 5-1 (2.3 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and phenyl isothiocyanate (0.9 ml). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-1. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 64.3%. ESI-MS (m/z, %) 484.29 (M−H)−; 1H NMR (400 MHz, DMSO-d6): δ9.63 (s, 1H), 9.21 (s, 1H), 9.11 (s, 1H), 7.85 (m, 4H), 7.73 (d, J=8.4 Hz, 2H), 7.34 (m, 2H), 7.00 (m, 1H), 4.94 (m, 1H), 2.88 (t, J=4.4 Hz, 4H), 2.42 (br, 4H), 2.19 (s, 3H), 1.70 (d, J=6.8 Hz, 6H) ppm.
Example 63
The Preparation of 4-(8-(3-chloro-4-fluorophenylamino)-9-isopropyl-9H-purin-2-ylamino)-N-(4-methylpiperidin-1-yl)benzamide




Compound 5-1 (2.3 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and 3-chloro-4-fluorophenyl isothiocyanate (1.35 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-1. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 62.9%. ESI-MS (m/z, %) 536.27 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.65 (s, 1H), 9.31 (s, 1H), 9.20 (s, 1H), 8.51 (s, 1H), 8.23 (m, 1H), 7.86 (d, J=8.8 Hz, 2H), 7.78 (m, 1H), 7.75 (t, J=6.6 Hz, 2H), 7.41 (t, J=9.2 Hz, 1H), 4.90 (s, 1H), 2.88 (d, J=4 Hz, 4H), 2.43 (m, 4H), 2.20 (s, 3H), 1.70 (d, J=6.8 Hz, 6H) ppm.
Example 64
The Preparation of 4-(8-(3-acetamidophenylamino)-9-isopropyl-9H-purin-2-ylamino)-N-(4-methylpiperidin-1-yl)benzamide




Compound 5-2 (2.4 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and meta-acetamidophenyl isothiocyanate (1.4 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-2. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 69.1%. ESI-MS (m/z, %) 561.18 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 10.00 (s, 1H), 9.52 (s, 1H), 9.20 (s, 1H), 8.88 (s, 1H), 8.42 (s, 1H), 8.26 (m, 1H), 8.08 (s, 1H), 7.66 (d, J=10.0 Hz, 2H), 7.57 (d, J=7.2 Hz, 1H), 7.21 (m, 2H), 4.94 (m, 1H), 2.90 (s, 4H), 2.68 (br, 4H), 2.06 (s, 3H), 1.64 (d, J=6.0 Hz, 6H) ppm.
Example 65
The Preparation of 3-fluoro-4-(9-isopropyl-8-phenylamino-9H-purin-2-ylamino)-N-(4-methylpiperazin-1-yl)benzamide




Compound 5-2 (2.4 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and phenyl isothiocyanate (0.9 ml). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-2. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 64.8%. ESI-MS (m/z, %) 502.17 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.37 (s, 1H), 9.10 (s, 1H), 8.88 (s, 1H), 8.43 (s, 1H), 8.25 (m, 1H), 7.84 (d, J=8.4 Hz, 2H), 7.64 (d, J=10.4 Hz, 2H), 7.00 (m, 2H), 4.90 (m, 1H), 2.89 (s, 4H), 2.42 (br, 4H), 2.19 (s, 3H), 1.65 (d, J=6.4 Hz, 6H) ppm.
Example 66
The Preparation of 3-fluoro-4-(9-isopropyl-8-(3-chloro-4-fluorophenylamino)-9H-purin-2-ylamino)-N-(4-methylpiperazin-1-yl)benzamide




Compound 5-2 (2.4 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and 3-chloro-4-fluorophenyl isothiocyanate (1.35 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-2. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 68.8%. ESI-MS (m/z, %) 554.30 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ9.21 (s, 1H), 9.04 (s, 1H), 8.41 (s, 1H), 8.22 (d, J=5.2 Hz, 1H), 7.74 (m, 1H), 7.62 (d, J=8.8 Hz, 1H), 7.57 (d, J=8.8 Hz, 1H), 7.39 (m, 1H), 6.87 (d, J=8.4 Hz, 2H), 4.84 (m, 1H), 3.05 (s, 4H), 2.45 (br, 4H), 2.22 (s, 3H), 1.66 (d, J=6.4 Hz, 6H) ppm.
Example 67
The Preparation of 3-fluoro-4-(9-isopropyl-8-phenylamino-9H-purin-2-ylamino)-N-(4-methylpiperazin-1-yl)benzamide




Compound 5-15 (2.4 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and phenyl isothiocyanate (0.9 ml). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-15. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 67.6%. ESI-MS (m/z, %) 502.22 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.22 (s, 1H), 8.87 (s, 1H), 8.49 (s, 1H), 8.44 (s, 1H), 8.29 (t, J=8.4, 1H), 7.86 (d, J=8.0, 2H), 7.78 (d, J=8.8, 2H), 7.34 (t, J=7.6, 2H), 7.01 (t, J=7.2, 1H), 4.97 (m, 1H), 4.04 (m, 1H), 3.38 (m, 2H), 3.09 (m, 2H), 2.72 (s, 3H), 1.99 (m, 4H), 1.65 (d, J=6.8, 6H) ppm.
Example 68
The Preparation of 3-methoxy-4-(9-isopropyl-8-phenylamino-9H-purin-2-ylamino)-N-(4-methylpiperazin-1-yl)benzamide




Compound 5-3 (2.5 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and phenyl isothiocyanate (0.9 ml). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-3. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 66.5%. ESI-MS (m/z, %) 514.21 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 10.98 (s, 1H), 9.65 (s, 1H), 8.90 (s, 1H), 8.50 (s, 1H), 8.27 (d, J=7.6 Hz, 1H), 7.83 (d, J=7.6 Hz, 2H), 7.39 (t, J=8 Hz, 2H), 7.18-7.08 (m, 3H), 5.026 (m, 1H), 3.94 (s, 4H), 3.42 (d, J=7.6 Hz, 2H), 3.09 (d, J=12 Hz, 2H), 2.79 (d, J=7.2 Hz, 3H), 1.66 (d, J=6.4 Hz, 6H) ppm.
Example 69
The Preparation of 4-(8-(3-chloro-4-fluorophenylamino)-9-isopropyl-9H-purin-2-ylamino)-3-methoxy-N-(1-methylpiperidin-4-yl)benzamide




To a solution of Compound 5-4 (2.0 g) and methyl 3-ethynylphenylcarbamodithioate (1.3 g) in N,N-dimethylformamide (30 ml) were added copper oxide (0.08 g) and potassium carbonate (1.4 g). The mixture was heated to 60° C. and reacted for 2-6 hours. The reaction solution was cooled to room temperature and filtered. The filtrate was washed with ethyl acetate, a saturated saline solution and water. The organic layer was dried over anhydrous sodium sulfate and concentrated. Purification was conducted by a column chromatography to obtain an off-white solid in a yield of 64.1%. ESI-MS (m/z, %) 507.22 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.51 (s, 1H), 9.04 (s, 1H), 8.91 (s, 1H), 7.88 (m, 4H), 7.65 (d, 2H), 7.41 (m, 2H), 7.12 (m, 1H), 4.80 (m, 1H), 4.14 (s, 1H), 3.71 (m, 1H), 2.80 (m, 4H), 2.39 (br, 4H), 2.21 (s, 3H), 1.62 (d, 6H) ppm.
Example 70
The Preparation of 4-(8-phenylamino-9-isopropyl-9H-purin-2-ylamino)-N-(1-methylpiperidin-4-yl)benzamide




Compound 5-4 (2.3 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and phenyl isothiocyanate (0.9 ml). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-4. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 67.9%. ESI-MS (m/z, %) 483.25 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.72 (s, 1H), 9.63 (s, 1H), 9.11 (s, 1H), 8.45 (s, 1H), 8.28 (s, 1H), 7.89-7.81 (m, 5H), 7.34 (t, J=7.8 Hz, 2H), 7.00 (t, J=7.2 Hz, 1H), 4.95 (m, 1H), 4.02 (s, 1H), 3.44 (d, J=10.8 Hz, 2H), 3.08 (s, 2H), 2.75 (s, 3H), 1.99 (s, 2H), 1.82 (d, J=11.2 Hz, 2H), 1.70 (d, J=6.8 Hz, 6H) ppm.
Example 71
The Preparation of 9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-8-phenylamino-9H-purine




Compound 5-7 (2.05 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and phenyl isothiocyanate (0.9 ml). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-7. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 67.5%. ESI-MS (m/z, %) 441.29 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.09 (d, J=6.8 Hz, 2H), 8.36 (s, 1H), 7.83 (d, J=8.0 Hz, 2H), 7.69 (d, J=9.2 Hz, 2H), 7.33 (m, 2H), 6.97 (m, 3H), 4.92 (m, 1H), 3.44 (br, 4H), 3.17 (br, 4H), 2.81 (s, 3H), 1.67 (d, J=6.8 Hz, 6H) ppm.
Example 72
The Preparation of 8-(3-bromophenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-7 (2.05 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and meta-bromophenyl isothiocyanate (1.5 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-7. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 57.4%. ESI-MS (m/z, %) 519.19 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.47 (s, 1H), 9.09 (s, 1H), 8.42 (s, 1H), 8.25 (s, 1H), 7.85 (d, J=8.0 Hz, 1H), 7.67 (d, J=8.8 Hz, 2H), 7.28 (t, J=8.4 Hz, 1H), 7.14 (d, J=8.0 Hz, 1H), 6.93 (d, J=8.8 Hz, 2H), 4.99 (m, 1H), 3.51-3.40 (m, 2H), 3.25 (s, 2H), 3.07 (s, 4H), 2.64 (s, 3H), 1.66 (d, J=6.8; H, 6H) ppm.
Example 73
The Preparation of 8-(3-ethynylphenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-7 (2.05 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and meta-ethynylphenyl isothiocyanate (1.1 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-7. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 53.9%. ESI-MS (m/z, %) 465.23 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.10 (s, 1H), 9.02 (s, 1H), 8.41 (s, 1H), 8.07 (s, 1H), 7.81 (d, J=8.4 Hz, 1H), 7.64 (d, J=9.2 Hz, 2H), 7.34 (t, J=8.0, 1H), 7.08 (d, J=7.2 Hz, 1H), 6.88 (d, J=9.2 Hz, 2H), 4.85 (m, 1H), 4.18 (s, 1H), 3.05 (s, 4H), 2.47 (t, J=9.0, 4H), 2.22 (s, 3H), 1.67 (d, J=6.8 Hz, 6H) ppm.
Example 74
The Preparation of 8-(3-chloro-4-fluorophenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-7 (2.05 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and 3-chloro-4-fluorophenyl isothiocyanate (1.35 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-7. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 60.8%. ESI-MS (m/z, %) 493.16 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.21 (s, 1H), 9.04 (s, 1H), 8.41 (s, 1H), 8.22 (d, J=5.2 Hz, 1H), 7.74 (m, 1H), 7.62 (d, J=8.8 Hz, 1H), 7.57 (d, J=8.8 Hz, 1H), 7.39 (m, 1H), 6.87 (d, J=8.4 Hz, 2H), 4.84 (m, 1H), 3.05 (br, 4H), 2.45 (br, 4H), 2.22 (s, 3H), 1.66 (d, J=6.4 Hz, 6H) ppm.
Example 75
The Preparation of 8-(3-bromophenylamino)-9-cyclopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-8 (2.05 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and meta-bromophenyl isothiocyanate (1.5 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-8. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 58.2%. ESI-MS (m/z, %) 517.21 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.78 (s, 1H), 9.32 (s, 1H), 8.45 (s, 1H), 7.79 (m, 2H), 7.61 (m, 2H), 7.49 (m, 2H), 7.36 (m, 1H), 4.15 (m, 1H), 2.94 (tr, 4H), 2.63 (br, 4H), 2.19 (s, 3H), 1.26 (m, 4H) ppm.
Example 76
The Preparation of 8-(4-bromophenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-7 (2.05 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and para-bromophenyl isothiocyanate (1.5 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-7. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 53.4%. ESI-MS (m/z, %) 519.22 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.25 (s, 1H), 9.11 (s, 1H), 8.39 (s, 1H), 7.85 (d, J=8.8 Hz, 2H), 7.69 (d, J=9.2 Hz, 2H), 7.50 (d, J=8.8 Hz, 2H), 6.95 (d, J=9.2 Hz, 2H), 4.91 (m, 1H), 3.69 (s, 1H), 3.45 (s, 1H), 3.17 (s, 1H), 3.00 (s, 1H), 2.82 (s, 3H), 1.66 (d, J=6.8 Hz, 6H) ppm.
Example 77
The Preparation of 8-(3-trifluoromethylphenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-7 (2.05 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and meta-trifluoromethylphenyl isothiocyanate (1.4 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-7. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 55.8%. ESI-MS (m/z, %) 509.24 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 10.80 (s, 1H), 10.03 (s, 1H), 8.50 (s, 1H), 8.32 (s, 1H), 8.19 (d, J=8.0, 1H), 7.62 (t, J=7.8, 1H), 7.55 (d, J=8.8, 2H), 7.40 (d, J=7.6, 1H), 7.04 (d, J=9.2, 2H), 5.05 (m, 1H), 3.78 (d, J=12.4, 4H), 3.04-3.20 (m, 4H), 2.82 (d, J=4.0, 3H), 1.64 (d, J=6.4, 6H) ppm.
Example 78
The Preparation of 8-(3-methoxyphenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-7 (2.05 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and meta-methoxyphenyl isothiocyanate (1.2 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-7. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 58.8%. ESI-MS (m/z, %) 471.28 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.06 (d, J=16.8, 2H), 8.37 (s, 1H), 7.69 (d, J=8.8, 2H), 7.52 (s, 1H), 7.42 (d, J=4.0, 1H), 7.22 (t, J=8.2, 1H), 6.95 (d, J=9.2, 2H), 6.56 (m, 1H), 4.91 (m, 1H), 3.76 (s, 1H), 2.81 (s, 1H), 1.66 (d, J=6.8, 6H) ppm.
Example 79
The Preparation of 8-(4-methoxyphenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-7 (2.05 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and para-methoxyphenyl isothiocyanate (1.2 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-7. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 59.8%. ESI-MS (m/z, %) 471.28 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.04 (s, 1H), 8.93 (s, 1H), 8.30 (s, 1H), 7.71 (m, J=8.9, 4H), 6.94 (t, J=9.4, 4H), 4.90 (m, 1H), 3.75 (s, 3H), 3.46-3.43 (m, 4H), 3.17-3.06 (m, 4H), 2.81 (s, 3H), 1.67 (d, J=6.8, 6H) ppm.
Example 80
The Preparation of 8-(3-chloro-4-(3-fluorobenzyloxy)phenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-7 (2.05 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and 3-chloro-4-(3-fluorobenzyloxy)phenyl isothiocyanate (2.1 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-7. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 65.7%. ESI-MS (m/z, %) 599.22 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.02 (d, J=4.8 Hz, 2H), 8.36 (s, 1H), 8.08 (d, J=2.4 Hz, 1H), 7.67 (m, 3H), 7.46 (m, 1H), 7.31 (t, J=7.6 Hz, 2H), 7.20 (m, 2H), 6.91 (d, J=8.8 Hz, 2H), 5.21 (s, 2H), 4.83 (m, 1H), 3.15 (s, 4H), 2.81 (s, 3H), 2.48 (s, 2H), 2.30 (s, 2H), 1.66 (d, J=6.8 Hz, 6H) ppm.
Example 81
The Preparation of 8-(3-chloro-4-((pyridin-2-yl)methoxy)phenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-7 (2.05 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and (3-chloro-4-(pyridin-2-yl)methoxy)phenyl isothiocyanate (2.0 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-7. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 65.9%. ESI-MS (m/z, %) 582.31 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.13 (s, 1H), 8.60 (d, 1H), 8.30 (s, 1H), 7.85 (m, 3H), 7.69 (m, 1H), 7.60 (d, 2H), 7.30-7.15 (m, 4H), 7.05 (d, 1H), 4.88 (m, 1H), 3.48 (br, 4H), 2.97 (br, 4H), 2.78 (s, 3H), 1.59 (d, J=6.8 Hz, 6H) ppm.
Example 82
The Preparation of 8-(3-(3-(3-chloro-4-fluorophenyl)ureido)phenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-7 (2.05 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and 3-(3-(3-chloro-4-fluorophenyl)ureido)phenyl isothiocyanate (2.3 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-7. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 62.5%. ESI-MS (m/z, %) 627.21 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 8.99 (d, J=9.6 Hz, 2H), 8.88 (s, 1H), 8.80 (s, 1H), 8.34 (s, 1H), 7.94 (s, 1H), 7.84 (m, 1H), 7.63 (d, J=9.2 Hz, 2H), 7.47 (d, J=8 Hz, 1H), 7.37-7.28 (m, 2H), 7.22 (t, J=8 Hz, 1H), 7.11 (d, J=8 Hz, 1H), 6.88 (d, J=9.2 Hz, 2H), 4.90 (m, 1H), 3.06 (t, J=4.8 Hz, 3H), 2.51 (m, 4H), 2.48 (d, J=4.4 Hz, 2H), 2.26 (d, J=19.6 Hz, 2H), 1.67 (d, J=6.8 Hz, 6H) ppm.
Example 83
The Preparation of 8-(4-(3-fluorophenylcarbamoyl)phenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-7 (2.05 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and 4-(3-fluorophenylcarbamoyl)phenyl isothiocyanate (2.0 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-7. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 54.6%. ESI-MS (m/z, %) 578.22 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 10.30 (s, 1H), 9.44 (s, 1H), 9.12 (s, 1H), 8.44 (s, 1H), 7.98 (s, 3H), 7.78 (d, J=11.6 Hz, 1H), 7.68 (d, J=8.8 Hz, 2H), 7.59 (d, J=7.2 Hz, 2H), 7.38 (m, 1H), 6.91 (m, 3H), 4.94 (m, 1H), 3.27 (s, 4H), 2.96 (d, J=8 Hz, 2H), 2.73 (s, 3H), 2.55 (s, 2H), 1.69 (d, J=6.4 Hz, 6H) ppm;
Example 84
The Preparation of 8-phenylamino-9-cyclopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-8 (2.0 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and phenyl isothiocyanate (0.86 ml). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-8. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 58.3%. ESI-MS (m/z, %) 439.23 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 10.14 (s, 1H), 9.64 (s, 1H), 8.39 (s, 1H), 7.851 (d, J=8 Hz, 2H), 7.60 (d, J=8.8 Hz, 2H), 7.41 (t, J=8 Hz, 2H), 7.13 (t, J=7.2 Hz, 1H), 7.05 (d, J=9.2 Hz, 2H), 3.43-3.50 (m, 4H), 3.20-3.24 (m, 4H), 2.82 (d, J=3.2 Hz, 3H), 1.19-1.30 (m, 4H) ppm.
Example 85
The Preparation of 8-phenylamino-9-methyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-6 (1.9 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and phenyl isothiocyanate (0.86 ml). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-6. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 51.1%. ESI-MS (m/z, %) 413.24 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.57 (s, 1H), 9.52 (s, 1H), 8.37 (s, 1H), 7.88 (d, J=7.6 Hz, 2H), 7.656 (d, J=8.4 Hz, 2H), 7.37 (t, J=8 Hz, 2H), 7.05 (t, J=7.2 Hz, 1H), 6.99 (d, J=8.8 Hz, 2H), 3.69 (s, 3H), 3.381 (s, 4H), 3.16 (d, J=8.8 Hz, 2H), 3.03 (d, J=12 Hz, 2H), 2.82 (s, 3H) ppm.
Example 86
The Preparation of 8-phenylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-5 (2.4 g) was dissolved into dichloromethane (120 ml). To the mixture were successively added EDCI (3.1 g), N,N-diisopropylethylamine (6.6 ml), and para-trifluoromethylphenyl isothiocyanate (1.1 ml). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-5. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 57.3%. ESI-MS (m/z, %) 399.27 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 10.89 (s, 1H), 9.93 (s, 1H), 8.77 (s, 1H), 8.20 (s, 1H), 7.30 (d, J=7.2 Hz, 2H), 7.63 (d, J=8.4 Hz, 2H), 7.34 (t, J=6.8 Hz, 2H), 7.00 (s, 1H), 6.87 (d, J=8.8 Hz, 2H), 3.04 (t, J=4.4 Hz, 4H), 2.46 (t, J=4.4 Hz, 4H), 2.23 (s, 3H) ppm.
Example 87
The Preparation of 8-phenylamino-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-9 (1.6 g) was dissolved into dichloromethane (65 ml). To the mixture were successively added EDCI (1.7 g), N,N-diisopropylethylamine (3.7 ml), and phenyl isothiocyanate (0.6 ml). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-9. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 52.1%. ESI-MS (m/z, %) 467.26 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.18 (s, 1H), 9.08 (s, 1H), 8.36 (s, 1H), 7.85 (d, J=7.6 Hz, 2H), 7.67 (d, J=8.4 Hz, 2H), 7.32 (t, J=3.6 Hz, 2H), 6.93-7.00 (m, 3H), 5.04 (t, J=8 Hz, 1H), 3.36 (s, 8H), 2.79 (s, 3H), 2.46 (s, 2H), 2.05 (s, 4H), 1.70 (s, 2H) ppm.
Example 88
The Preparation of 8-(4-trifluoromethylphenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-7 (2.05 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and para-trifluoromethylphenyl isothiocyanate (1.4 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-7. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 55.4%. ESI-MS (m/z, %) 509.25 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.46 (s, 1H), 9.09 (s, 1H), 8.44 (s, 1H), 8.03 (d, J=8.4 Hz, 2H), 7.67 (m, J=8.8 Hz, 4H), 6.90 (d, J=8.8 Hz, 2H), 4.90 (m, 1H), 3.10 (s, 4H), 2.59 (s, 4H), 2.32 (s, 3H), 1.68 (d, J=6.4 Hz, 6H) ppm.
Example 89
The Preparation of 8-(3-acrylylaminophenylamino)-9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-7 (2.05 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and 3-acrylylaminophenyl isothiocyanate (1.5 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-7. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 58.9%. ESI-MS (m/z, %) 510.25 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.51 (s, 1H), 9.07 (s, 1H), 8.52 (s, 1H), 8.09 (d, J=8.4 Hz, 2H), 7.61 (m, 4H), 6.92 (d, J=8.8 Hz, 2H), 6.54 (s, 1H), 6.05 (s, 1H), 5.59 (s, 1H), 4.79 (m, 1H), 2.93 (s, 4H), 2.62 (br, 4H), 2.38 (s, 3H), 1.61 (d, J=6.4 Hz, 6H) ppm.
Example 90
The Preparation of 9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-8-(pyridin-3-yl)-9H-purine




Compound 5-7 (2.05 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and 3-pyridinyl isothiocyanate (1.0 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-7. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 65.7%. ESI-MS (m/z, %) 442.26 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.38 (s, 1H), 9.13 (s, 1H), 8.99 (s, 1H), 8.40 (s, 1H), 8.36 (d, J=8.4 Hz, 1H), 8.20 (d, J=4.4 Hz, 1H), 7.70 (d, J=8.8 Hz, 2H), 7.37 (m, 1H), 6.96 (d, J=8.8 Hz, 2H), 4.97-4.92 (m, 1H), 3.35 (s, 6H), 2.80 (s, 3H), 2.53 (s, 2H), 1.69 (s, 6H) ppm.
Example 91
The Preparation of 9-isopropyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-8-cyclohexyl-9H-purine




Compound 5-7 (2.05 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and cyclohexyl isothiocyanate (1.0 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-7. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 67.8%. ESI-MS (m/z, %) 447.28 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 10.89 (s, 1H), 9.26 (s, 1H), 8.23 (s, 1H), 7.62 (d, J=9.2, 2H), 6.95 (d, J=8.8, 2H), 4.72 (m, 1H), 4.03 (m, J=7.1, 1H), 3.69 (s, 4H), 3.13-3.09 (m, 4H), 2.80 (s, 3H), 1.98 (d, J=5.6, 2H), 1.76 (d, J=9.6, 2H), 1.57 (d, J=6.8, 6H), 1.37-1.26 (m, 6H) ppm.
Example 92
The Preparation of 9-cyclopentyl-6-methyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-8-phenyl-9H-purine




Compound 5-10 (2.3 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and phenyl isothiocyanate (0.9 mlg). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-10. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 61.1%. ESI-MS (m/z, %) 481.27 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.12 (s, 1H), 8.39 (s, 1H), 7.75 (m, 2H), 7.59 (m, 2H), 7.46 (m, 2H), 7.10 m, 3H), 4.62 (m, 1H), 3.38 (s, 6H), 2.75 (s, 3H), 2.68 (s, 3H), 2.42 (br, 2H), 2.15 (s, 4H), 1.76 (m, 4H) ppm.
Example 93
The Preparation of 9-isopropyl-6-methoxy-2-(4-(4-methylpiperazin-1-yl)phenylamino)-8-phenyl-9H-purine




Compound 5-11 (2.2 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and phenyl isothiocyanate (0.9 mlg). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-11. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 63.5%. ESI-MS (m/z, %) 471.28 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 8.56 (s, 1H), 7.82 (d, J=8.0 Hz, 2H), 7.61 (d, J=8.4 Hz, 2H), 7.39 (m, 2H), 7.26 (m, 2H), 7.02 (m, 2H), 4.92 (m, 1H), 4.06 (s, 3H), 3.49 (br, 4H), 3.27 (br, 4H), 2.83 (s, 3H), 1.61 (d, J=6.8 Hz, 6H) ppm.
Example 94
The Preparation of 9-isopropyl-6-methylamino-2-(4-(4-methylpiperazin-1-yl)phenylamino)-8-phenyl-9H-purine




Compound 5-12 (2.2 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and phenyl isothiocyanate (0.9 mlg). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-12. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 65.4%. ESI-MS (m/z, %) 470.25 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 8.61 (s, 1H), 7.83 (d, J=8.0 Hz, 2H), 7.69 (m, 2H), 7.37 (m, 2H), 7.25 (m, 1H), 7.04 (m, 3H), 4.71 (m, 1H), 3.49 (br, 4H), 3.27 (br, 4H), 2.86 (s, 3H), 1.65 (d, J=6.8 Hz, 6H) ppm.
Example 95
The Preparation of 9-isopropyl-6-methyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-8-phenyl-9H-purine




Compound 5-13 (2.2 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and phenyl isothiocyanate (0.9 ml). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-13. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 66.7%. ESI-MS (m/z, %) 455.28 (M−H)−; 1H NMR (400 MHz, DMSO-d6): δ 9.08 (s, 1H), 9.01 (s, 1H), 7.84 (d, J=8.0, 2H), 7.71 (d, J=8.8, 2H), 7.32 (t, J=7.8, 2H), 6.96 (t, J=8.4, 3H), 4.91 (m, J=6.6, 1H), 3.67 (s, 4H), 3.27 (s, 4H), 2.80 (s, 3H), 2.49 (s, 3H), 1.66 (d, J=6.8, 6H) ppm.
Example 96
The Preparation of 8-(3-chloro-4-fluorophenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-9 (2.2 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and 3-chloro-4-fluorophenyl isothiocyanate (1.35 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-9. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 67.6%. ESI-MS (m/z, %) 519.20 (M−H)−; 1H NMR (400 MHz, DMSO-d6): δ9.41 (s, 1H), 9.10 (s, 1H), 8.42 (s, 1H), 8.23 (m, J=3.07, 1H), 7.80 (m, 1H), 7.66 (d, J=8.8, 2H), 7.39 (t, J=9.2, 1H), 6.94 (d, J=9.2, 2H), 5.01 (m, 1H), 3.41 (s, 4H), 2.81 (s, 3H), 2.54 (s, 4H), 2.46 (s, 2H), 2.05 (s, 4H), 1.70 (d, J=4.4, 2H) ppm.
Example 97
The Preparation of 9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-8-(pyridin-3-yl)-9H-purine




Compound 5-9 (2.2 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and 3-pyridinyl isothiocyanate (1.0 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-9. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 64.3%. ESI-MS (m/z,%) 468.23 (M−H)−; 1H NMR (400 MHz, DMSO-d6): δ 9.40 (s, 1H), 9.11 (s, 1H), 8.97 (d, J=2.0, 1H), 8.40 (s, 1H), 8.35 (d, J=8.4, 1H), 8.20 (d, J=4.4, 1H), 7.66 (d, J=8.8, 2H), 7.36 (m, J=6.6, 1H), 6.93 (d, J=8.8, 2H), 5.02 (m, 1H), 3.20 (s, 4H), 2.73 (s, 3H), 2.47 (s, 4H), 2.06 (s, 4H), 1.71 (s, 2H), 1.23 (s, 2H) ppm.
Example 98
The Preparation of 4-(9-cyclopentylamino-8-(pyridin-3-ylamino)-9H-purin-2-ylamino)-N-(4-methylpiperidin-1-yl)benzamide




Compound 5-14 (2.46 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and 3-pyridinyl isothiocyanate (1.0 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-14. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 70.3%. ESI-MS (m/z, %) 511.23 (M−H)−; 1H NMR (400 MHz, DMSO-d6): δ 9.66 (d, J=11.2, 2H), 9.48 (s, 1H), 8.99 (d, J=2.0, 1H), 8.49 (s, 1H), 8.36 (d, J=7.6, 1H), 8.22 (d, J=4.0, 1H), 7.86 (d, J=8.8, 2H), 7.75 (d, J=8.4, 2H), 7.39 (m, J=4.3, 1H), 5.06 (m, 1H), 3.21 (s, 4H), 2.78 (s, 3H), 2.51 (s, 4H), 2.10 (s, 4H), 1.76 (s, 2H), 1.24 (s, 2H) ppm.
Example 99
The Preparation of N-(3-(9-isopropyl-8-phenylamino-9H-purin-2-ylamino)phenyl)acrylamide




Compound 5-16 (1.87 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and phenyl isothiocyanate (0.9 ml). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-16. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 65.5%. ESI-MS (m/z, %) 414.23 (M−H)+; 1H NMR (400 MHz, DMSO-d6): δ 10.05 (s, 1H), 9.29 (s, 1H), 9.02 (s, 1H), 8.40 (s, 1H), 8.02 (s, 1H), 7.84 (d, J=8.0, 2H), 7.57 (d, J=6.8, 1H), 7.34 (t, J=7.2, 2H), 7.20 (t, J=7.4, 2H), 7.00 (t, J=7.2, 1H), 6.49 (m, J=9.1, 1H), 6.26 (d, J=16.8, 1H), 5.74 (d, J=10.0, 1H), 4.90 (m, 1H), 1.68 (d, J=6.8, 6H) ppm.
Example 100
The Preparation of 9-isopropylamino-2-(4-(2-morpholinoethoxy)phenylamino))-8-phenylamino-9H-purine




Compound 5-17 (2.23 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and phenyl isothiocyanate (0.9 ml). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-17. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 70.5%. ESI-MS (m/z, %) 474.26 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.08 (s, 1H), 8.30 (s, 1H), 8.01 (s, 1H), 7.88 (d, J=8.0 Hz, 2H), 7.71 (d, J=9.2 Hz, 2H), 7.30 (m, 2H), 7.17 (m, 3H), 4.84 (m, 1H), 4.04 (m, 2H), 3.84 (m, 4H), 3.24 (br, 4H), 2.81 (m, 2H), 1.63 (d, J=6.8 Hz, 6H) ppm.
Example 101
The Preparation of 9-isopropylamino-2-(4-(3-(4-methylpiperazin-1-yl)propylamino)phenylamino)-8-phenylamino-9H-purine




Compound 5-18 (2.39 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and phenyl isothiocyanate (0.9 ml). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-18. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 69.1%. ESI-MS (m/z, %) 500.29 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.09 (s, 1H), 8.56 (s, 1H), 7.83 (s, 1H), 7.69 (s, 1H), 7.53 (m, 2H), 7.36 (m, 4H), 6.96 (m, 3H), 4.95 (m, 1H), 3.44 (m, 2H), 3.11 (m, 2H), 2.95 (m, 8H), 2.41 (s, 3H), 1.68 (m, 2H), 1.62 (d, J=6.8 Hz, 6H) ppm.
Example 102
The Preparation of 8-(6-chloropyrimidin-3-yl)-9-isopropylamino-2-(4-(3-(4-methylpiperazin-1-yl)propylamino)phenylamino)-9H-purine




Compound 5-7 (2.05 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and 2-chloro-5-pyridinyl isothiocyanate (1.3 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-7. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 69.8%. ESI-MS (m/z, %) 476.29 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.65 (s, 1H), 9.21 (s, 1H), 8.85 (s, 1H), 8.44 (d, J=6.8 Hz, 2H), 7.68 (d, J=8.0 Hz, 2H), 7.49 (d, J=8.8 Hz, 1H), 6.96 (d, J=8.0 Hz, 2H), 4.96 (m, 1H), 3.71 (m, 2H), 3.47 (m, 2H), 3.15 (br, 2H), 3.03 (m, 2H), 2.82 (s, 2H), 1.67 (d, J=6.4 Hz, 6H) ppm.
Example 103
The Preparation of 8-(3-chloro-4-(3-fluorobenzyloxy)phenylamino)-9-isopropyl-2-(4-morpholinophenylamino)-9H-purine




Compound 5-19 (1.97 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and phenyl isothiocyanate (0.9 ml). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-19. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale yellow solid in a yield of 70.6%. ESI-MS (m/z, %) 586.27 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ 9.01 (d, J=7.2 Hz, 2H), 8.37 (s, 1H), 8.08 (d, J=2.4 Hz, 1H), 7.67 (m, 3H), 7.46 (m, 1H), 7.31 (m, 2H), 7.20 (m, 2H), 6.91 (d, J=9.2 Hz, 2H), 5.21 (s, 2H), 4.83 (m, 1H), 3.74 (m, 4H), 3.03 (m, 4H), 1.67 (d, J=6.4 Hz, 6H) ppm.
Example 104
The Preparation of 8-(4-bromophenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-9 (2.2 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and 4-bromophenyl isothiocyanate (1.65 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-9. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 66.7%. ESI-MS (m/z, %) 548.18 (M−H)+; 1H NMR (400 MHz, DMSO-d6): δ9.22 (s, 1H), 9.09 (s, 1H), 8.39 (s, 1H), 7.83 (d, J=9.2 Hz, 2H), 7.65 (d, J=7.0 Hz, 2H), 7.50 (d, J=9.2 Hz, 2H), 6.93 (d, J=8.8 Hz, 2H), 4.95 (m, 1H), 3.27-3.02 (m, 4H), 2.75 (m, 4H), 2.45 (s, 3H), 2.05 (m, 4H), 1.68 (m, 2H), 1.23 (m, 2H).
Example 105
The Preparation of 8-(3-nitrophenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-9 (2.2 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and 3-nitrophenyl isothiocyanate (1.50 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-9. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 65.4%. ESI-MS (m/z, %) 514.23 (M−H)+; 1H NMR (400 MHz, DMSO-d6): δ9.75 (s, 1H), 9.23 (s, 1H), 8.87 (s, 1H), 8.45 (s, 1H), 8.32 (d, J=7.6 Hz, 1H), 7.84 (d, J=7.2 Hz, 1H), 7.65 (m, 3H), 6.95 (d, J=8.8 Hz, 2H), 5.04 (m, 1H), 3.72 (m, 2H), 3.49 (m, 2H), 3.16 (m, 2H), 2.99 (m, 2H), 2.83 (s, 3H), 2.45 (m, 2H), 2.06 (m, 4H), 1.71 (m, 2H).
Example 106
The Preparation of 8-benzylamino-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-9 (2.2 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and benzyl isothiocyanate (1.45 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-9. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 63.9%. ESI-MS (m/z, %) 483.26 (M−H)+; 1H NMR (400 MHz, DMSO-d6): δ8.97 (s, 1H), 8.15 (s, 1H), 7.63 (m, 3H), 7.35 (m, 4H), 7.25 (m, 1H), 6.91 (d, J=9.2 Hz, 2H), 4.75 (m, 1H), 4.56 (d, J=8.0 Hz, 2H), 3.67 (m, 2H), 3.44 (m, 2H), 3.16 (m, 2H), 2.97 (m, 2H), 2.82 (s, 3H), 2.38 (m, 2H), 1.99 (m, 4H), 1.65 (m, 2H).
Example 107
The Preparation of 8-(3-ethynylphenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-9 (2.2 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and 3-ethynylphenyl isothiocyanate (1.34 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-9. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 69.5%. ESI-MS (m/z, %) 493.28 (M−H)+; 1H NMR (400 MHz, DMSO-d6): δ9.26 (s, 1H), 9.13 (s, 1H), 8.43 (s, 1H), 8.09 (s, 1H), 7.84 (d, J=8.0 Hz, 1H), 7.67 (d, J=8.4 Hz, 2H), 7.34 (m, 1H), 7.09 (d, J=7.2 Hz, 1H), 6.94 (d, J=10.8 Hz, 2H), 5.00 (m, 1H), 4.20 (s, 1H), 3.67 (m, 2H), 3.47 (m, 2H), 3.17 (m, 2H), 3.00 (m, 2H), 2.78 (s, 3H), 2.46 (m, 2H), 2.05 (m, 4H), 1.71 (m, 2H).
Example 108
The Preparation of 8-(2-fluoro-4-bromophenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-9 (2.2 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and 2-fluoro-4-bromophenyl isothiocyanate (1.60 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-9. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 69.7%. ESI-MS (m/z, %) 565.20 (M−H)+; 1H NMR (400 MHz, DMSO-d6): δ9.06 (s, 1H), 8.93 (s, 1H), 8.35 (s, 1H), 7.85 (m, 1H), 7.61 (m, 3H), 7.40 (d, J=8.4 Hz, 1H), 8.67 (d, J=9.2 Hz, 2H), 4.88 (m, 1H), 3.10 (m, 4H), 2.62 (m, 4H), 2.43 (m, 2H), 2.34 (s, 3H), 2.03 (m, 4H), 1.68 (m, 2H).
Example 109
The Preparation of 8-phenylamino-9-cyclopentyl-2-(4-(4-morpholinylmethyl)phenylamino)-9H-purine




Compound 5-20 (2.2 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and phenyl isothiocyanate (1.0 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-9. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 69.9%. ESI-MS (m/z, %) 470.27 (M−H)+. 1H NMR (400 MHz, DMSO-d6): δ9.57 (s, 1H), 9.16 (s, 1H), 8.45 (s, 1H), 7.91 (s, 1H), 7.86 (d, J=8.0 Hz, 2H), 7.76 (s, 1H), 7.47 (s, 1H), 7.38 (m, 2H), 7.22 (s, 1H), 7.03 (m, 1H), 5.04 (m, 1H), 4.10 (m, 2H), 3.70 (m, 4H), 3.15 (m, 2H), 2.37 (m, 2H), 2.05 (m, 4H), 1.75 (m, 2H), 1.29 (m, 2H).
Example 110
The Preparation of 8-(3-fluorophenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-9 (2.2 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (4.9 ml), and 3-fluorophenyl isothiocyanate (1.0 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-9. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 67.4%. ESI-MS (m/z, %) 487.25 (M−H)+; 1H NMR (400 MHz, DMSO-d6): δ9.42 (s, 1H), 9.13 (s, 1H), 8.42 (s, 1H), 7.92 (d, J=12.0 Hz, 1H), 7.67 (d, J=9.2 Hz, 2H), 7.59 (d, J=8.0 Hz, 1H), 7.35 (m, 1H), 6.94 (d, J=8.8 Hz, 2H), 6.79 (m, 1H), 5.02 (m, 1H), 3.68 (m, 2H), 3.44 (m, 2H), 3.18 (m, 2H), 3.01 (m, 2H), 2.81 (s, 3H), 2.44 (m, 2H), 2.05 (m, 4H), 1.70 (m, 2H).
Example 111
The Preparation of 8-phenylamino-9-cyclohexyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-21 (1.53 g) was dissolved into dichloromethane (35 ml). To the mixture were successively added EDCI (1.54 g), N,N-diisopropylethylamine (3.31 ml), and phenyl isothiocyanate (0.56 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-21. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 69.4%. ESI-MS (m/z, %) 483.25 (M−H)+. 1H NMR (400 MHz, DMSO-d6): δ9.04 (s, 1H), 8.99 (s, 1H), 8.34 (s, 1H), 7.80 (d, J=8.4 Hz, 2H), 7.67 (d, J=8.8 Hz, 2H), 7.33 (m, 2H), 6.98 (m, 1H), 6.87 (d, J=8.4 Hz, 2H), 4.74 (m, 1H), 3.05 (m, 4H), 2.61 (m, 2H), 5.47 (m, 4H), 2.23 (s, 3H), 1.91 (m, 2H), 1.81 (m, 3H), 1.44 (m, 2H), 1.34 (m, 1H).
Example 112
The Preparation of 8-(3-hydroxyphenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-9 (2.1 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine-2.1 ml), and 3-hydroxyphenyl isothiocyanate (1.05 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-9. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 70.4%. ESI-MS (m/z, %) 485.25 (M−H)+; 1H NMR (400 MHz, DMSO-d6): δ9.36 (s, 1H), 9.07 (s, 1H), 8.93 (s, 1H), 8.35 (s, 1H), 7.67 (d, J=9.2 Hz, 2H), 7.38 (s, 1H), 7.16 (d, J=9.2 Hz, 1H), 7.08 (m, 1H), 6.93 (d, J=9.2 Hz, 2H), 6.39 (s, 1H), 4.96 (m, 1H), 3.43 (m, 2H), 3.34 (m, 2H), 3.16 (m, 4H), 2.75 (s, 1H), 2.46 (m, 2H), 2.04 (m, 4H), 1.69 (m, 2H).
Example 113
The Preparation of 8-(3-chlorophenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-9 (2.2 g) was dissolved into dichloromethane (70 ml). To the mixture were successively added EDCI (2.7 g), N,N-diisopropylethylamine (2.5 ml), and 3-chlorophenyl isothiocyanate (1.42 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-9. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 71.7%. ESI-MS (m/z, %) 503.25 (M−H)+; 1H NMR (400 MHz, DMSO-d6): δ9.38 (s, 1H), 9.13 (s, 1H), 8.44 (s, 1H), 8.11 (s, 1H), 7.75 (d, J=8.0 Hz, 1H), 7.65 (d, J=8.4 Hz, 2H), 7.35 (m, 1H), 7.02 (m, 1H), 6.94 (d, J=8.8 Hz, 2H), 5.01 (m, 1H), 3.43 (m, 4H), 3.10 (m, 4H), 2.82 (s, 3H), 2.45 (m, 2H), 2.05 (m, 4H), 1.70 (m, 2H) ppm.
Example 114
The Preparation of 8-(3-methylphenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-9 (2.1 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine 2.1 ml), and 3-methylphenyl isothiocyanate (1.0 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-9. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 72.7%. ESI-MS (m/z, %) 483.25 (M−H)+; 1H NMR (400 MHz, DMSO-d6): δ9.08 (s, 1H), 9.02 (s, 1H), 8.37 (s, 1H), 7.68-7.62 (m, 4H), 7.22 (m, 1H), 6.93 (d, J=8.8 Hz, 2H), 6.80 (d, J=7.6 Hz, 1H), 4.99 (m, 1H), 3.67 (m, 2H), 3.44 (m, 2H), 3.03 (m, 4H), 2.81 (s, 3H), 2.45 (m, 2H), 2.31 (s, 3H), 2.04 (m, 4H), 1.69 (m, 2H) ppm.
Example 115
The Preparation of 8-(3,5-dichlorophenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-9 (2.55 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (2.67 g), N,N-diisopropylethylamine 2.7 ml), and 3,5-dichlorophenyl isothiocyanate (1.7 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-9. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 73.7%. ESI-MS (m/z, %) 537.25 (M−H)+; 1H NMR (400 MHz, DMSO-d6): δ9.40 (s, 1H), 9.08 (s, 1H), 8.48 (s, 1H), 7.97 (s, 1H), 7.60 (d, J=8.8 Hz, 2H), 7.21 (m, 2H), 6.99 (m, 2H), 4.90 (m, 1H), 3.12 (m, 2H), 3.04 (m, 2H), 2.54 (m, 4H), 2.21 (m, 4H), 2.04 (s, 3H), 1.67 (m, 2H), 1.19 (m, 2H) ppm.
Example 116
The Preparation of 8-(2,5-difluorophenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-9 (2.7 g) was dissolved into dichloromethane (100 ml). To the mixture were successively added EDCI (2.8 g), N,N-diisopropylethylamine (3 ml), and 2,5-difluorophenyl isothiocyanate (1.5 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-9. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 70.6%. ESI-MS (m/z, %) 505.25 (M−H)+; 1H NMR (400 MHz, DMSO-d6): δ9.16 (s, 1H), 9.06 (s, 1H), 8.43 (s, 1H), 7.91 (m, 1H), 7.56 (m, 2H), 7.32 (m, 1H), 6.91 (m, 3H), 4.92 (m, 1H), 3.45 (m, 4H), 3.23-3.03 (m, 4H), 2.80 (s, 3H), 2.42 (m, 2H), 2.03 (m, 4H), 1.68 (m, 2H) ppm.
Example 117
The Preparation of 8-phenylamino-9-cyclopentyl-2-(4-(2-methoxyethoxy)phenylamino)-9H-purine




Compound 5-22 (3.2 g) was dissolved into dichloromethane (90 ml). To the mixture were successively added EDCI (3.6 g), N,N-diisopropylethylamine 3.3 ml), and phenyl isothiocyanate (1.52 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-9. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 75.3%. ESI-MS (m/z, %) 443.25 (M−H)−. 1H NMR (400 MHz, DMSO-d6): δ9.20 (s, 1H), 9.06 (s, 1H), 8.36 (s, 1H), 7.86 (d, J=9.6 Hz, 2H), 7.65 (d, J=8.0 Hz, 2H), 7.34 (m, 2H), 6.97 (m, 1H), 6.85 (d, J=8.0 Hz, 2H), 5.05 (m, 1H), 4.04 (m, 2H), 3.64 (m, 2H), 3.31 (s, 3H), 2.43 (m, 2H), 2.03 (m, 4H), 1.68 (m, 2H) ppm.
Example 118
The Preparation of 8-(2,4,5-trichlorophenylamino)-9-cyclopentyl-2-(4-(4-methyl piperazin-1-yl)phenylamino)-9H-purine




Compound 5-9 (2.1 g) was dissolved into dichloromethane (60 ml). To the mixture were successively added EDCI (2.15 g), N,N-diisopropylethylamine (2 ml), and 2,4,5-trichlorophenyl isothiocyanate (1.6 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-9. After cooling the mixture, a solid was separated out and purified by recrystallization to obtain a pale yellow solid in a yield of 74.3%. ESI-MS (m/z, %) 571.25 (M−H)+; 1H NMR (400 MHz, DMSO-d6): δ9.09 (s, 1H), 9.01 (s, 1H), 8.27 (s, 1H), 7.64 (m, 2H), 7.12 (m, 1H), 6.89 (d, J=8.4 Hz, 2H), 6.78 (d, J=7.2 Hz, 1H), 4.93 (m, 1H), 3.64 (m, 2H), 3.41 (m, 2H), 3.13 (m, 4H), 2.75 (s, 3H), 2.43 (m, 2H), 2.14 (m, 4H), 1.65 (m, 2H) ppm.
Example 119
The Preparation of 8-phenylamino-9-cyclopentyl-2-(4-((4-ethylpiperazin-1-yl)methyl)phenylamino)-9H-purine




Compound 5-23 (4.6 g) was dissolved into dichloromethane (100 ml). To the mixture were successively added EDCI (1.67 g), N,N-diisopropylethylamine 1.2 ml), and phenyl isothiocyanate (0.79 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-25. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 65.1%. ESI-MS (m/z, %) 497.25 (M−H)+. 1H NMR (400 MHz, DMSO-d6): δ9.25 (s, 1H), 9.06 (s, 1H), 8.40 (s, 1H), 7.82 (d, J=8.0 Hz, 2H), 7.71 (d, J=8.4 Hz, 2H), 7.34 (m, 2H), 7.15 (d, J=8.4 Hz, 2H), 6.99 (m, 1H), 4.97 (m, 1H), 3.38 (s, 2H), 2.47-2.29 (m, 12H), 2.05 (m, 2H), 1.71 (m, 2H), 0.97 (m, 3H) ppm.
Example 120
The Preparation of 8-phenylamino-9-cyclopentyl-2-(4-((4-methylpiperazin-1-yl)methyl)phenylamino)-9H-purine




Compound 5-25 (4.6 g) was dissolved into dichloromethane (100 ml). To the mixture were successively added EDCI (1.74 g), N,N-diisopropylethylamine 1.2 ml), and phenyl isothiocyanate (0.82 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-25. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 71.8%. ESI-MS (m/z, %) 483.25 (M−H)+. 1H NMR (400 MHz, DMSO-d6): δ9.25 (s, 1H), 9.05 (s, 1H), 8.39 (s, 1H), 7.82 (d, J=8.0 Hz, 2H), 7.70 (d, J=8.0 Hz, 2H), 7.33 (m, 2H), 7.15 (d, J=8.0 Hz, 2H), 6.99 (m, 1H), 4.97 (m, 2H), 3.38 (m, 4H), 2.50 (m, 4H), 2.33 (m, 4H), 2.14 (s, 3H), 2.05 (m, 4H), 1.71 (m, 2H) ppm.
Example 121
The Preparation of 8-(2,5-dichlorophenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-9 (1.3 g) was dissolved into dichloromethane (60 ml). To the mixture were successively added EDCI (0.8 g), N,N-diisopropylethylamine (2 ml), and 2,5-dichlorophenyl isothiocyanate (0.72 g). The mixture was stirred at room temperature for half an hour, and refluxed for 16 hours. TLC detection indicated the completion of the reaction of the starting material 5-9. Purification was conducted by a column chromatography to obtain a pale yellow solid in a yield of 64.3%. ESI-MS (m/z, %) 537.23 (M−H)+. 1H NMR (400 MHz, DMSO-d6): δ9.02 (s, 1H), 8.62 (s, 1H), 8.37 (s, 1H), 7.91 (s, 1H), 7.59 (d, J=8.4 Hz, 2H), 7.49 (d, J=7.6 Hz, 1H), 7.10 (m, 1H), 6.86 (d, J=9.2 Hz, 2H), 4.91 (m, 1H), 3.04 (m, 4H), 2.45 (m, 4H), 2.22 (s, 3H), 2.00 (m, 4H), 1.67 (m, 2H) ppm.
Example 122
The Preparation of 8-(2,6-dichlorophenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-9 (2.2 g) was dissolved into dichloromethane (40 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (5 ml), and 2,4-dichlorophenyl isothiocyanate (1.5 g). The mixture was stirred at room temperature for half an hour, and refluxed for 12 hours. TLC detection indicated the completion of the reaction of the starting material 5-9. Purification was conducted by a column chromatography to obtain a pale yellow solid in a yield of 67.4%. ESI-MS (m/z, %) 537.22 (M−H)+. 1H NMR (400 MHz, DMSO-d6): δ9.01 (s, 1H), 8.70 (s, 1H), 8.26 (s, 1H), 7.59 (d, J=8.4 Hz, 4H), 7.39 (s, 1H), 6.86 (d, J=8.8 Hz, 2H), 4.90 (m, 1H), 3.04 (m, 4H), 2.45 (m, 6H), 2.22 (s, 3H), 2.00 (m, 4H), 1.67 (m, 2H) ppm.
Example 123
The Preparation of 8-(3-bromophenylamino)-9-cyclopentyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-9 (2.2 g) was dissolved into dichloromethane (40 ml). To the mixture were successively added EDCI (2.3 g), N,N-diisopropylethylamine (5 ml), and 3-bromophenyl isothiocyanate (1.54 g). The mixture was stirred at room temperature for half an hour, and refluxed for 12 hours. TLC detection indicated the completion of the reaction of the starting material 5-9. Purification was conducted by a column chromatography to obtain a pale yellow solid in a yield of 69.5%. ESI-MS (m/z, %) 547.12 (M−H)+. 1H NMR (400 MHz, DMSO-d6): δ9.34 (s, 1H), 9.13 (s, 1H), 8.44 (s, 1H), 8.22 (s, 1H), 7.83 (d, J=8.4 Hz, 1H), 7.67 (d, J=8.8 Hz, 2H), 7.29 (m, 1H), 7.15 (d, J=8.0 Hz, 1H), 6.94 (d, J=9.2 Hz, 2H), 5.01 (m, 1H), 3.65 (m, 4H), 3.28 (m, 4H), 2.81 (s, 3H), 2.45 (m, 2H), 2.05 (m, 4H), 1.70 (m, 2H) ppm.
Example 124
The Preparation of 8-phenylamino-9-cyclohexylmethyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)-9H-purine




Compound 5-23 (2.0 g) was dissolved into dichloromethane (25 ml). To the mixture were successively added EDCI (1.92 g), N,N-diisopropylethylamine (2.25 ml), and phenyl isothiocyanate (0.82 g). The mixture was stirred at room temperature for half an hour, and refluxed for 10 hours. TLC detection indicated the completion of the reaction of the starting material 5-23. After cooling the mixture, purification was conducted by a column chromatography to obtain a pale red solid in a yield of 66.2%. ESI-MS (m/z, %) 497.26 (M−H)+. 1H NMR (400 MHz, DMSO-d6): δ9.04 (s, 1H), 8.59 (s, 1H), 8.30 (s, 1H), 7.81 (d, J=8.4 Hz, 2H), 7.65 (d, J=8.8 Hz, 2H), 7.31 (m, 2H), 6.88 (m, 1H), 6.75 (d, J=8.4 Hz, 2H), 3.78 (m, 2H), 3.05 (m, 4H), 1.82-1.54 (m, 7H), 1.36-1.18 (m, 4H), 1.12-1.00 (m, 2H).
Assay
Biological Assessment
Assay 1: The Test of the Arylamino Purine Derivatives for the Kinase Inhibitory Activity
The object of this assay was to test the inventive compounds for the kinase inhibitory activity in vitro. In this assay, an isotopic labeling method was used to label the γ phosphate group on ATP. EGFR (including wild type, L858R mutant type and L858R/T790M double mutant type), VEGFR2, ALK, BTK, c-KIT, c-SRC, MET, PDGFRα and FLT3 kinases were tested in vitro for the activity inhibition. Staurosporine was used as a reference molecule (or referred to as a positive control). The kinase inhibitory activities of the tested compounds were expressed in the IC50 value (half inhibition concentration) or the kinase activity inhibitory rate by the tested compounds at 10 μM. The IC50 value can be obtained by the calculation of the inhibitory rates at a series of different concentrations of the tested compounds.
1. Materials
20 mM 3-(N-morpholinyl)propylsulfonic acid (MOPS);
1 mM Ethylenediaminetetraacetic acid (EDTA);
0.01% Polyethylene glycol lauryl ether (Brij-35);
5% Glycerol;
0.1% Mercaptoethanol;
1 mg/ml Bovine serum albumin (BSA);
10 mM Manganous dichloride solution (MnCl2);
0.1 mg/ml Glutamic acid/tyrosine (4:1) polymerized polypeptide (poly(Glu,Tyr)4:1) (the substrate for wild type and L858R mono mutant type EGFRs, c-KIT and PDGFRα);
250 μM polypeptide GGMEDIYFEFMGGKKK (the substrate for L858R/T790M double mutant type EGFR);
250 μM polypeptide KKKSPGEYVNIEFG (the substrate for ALK and MET);
250 μM polypeptide KVEKIGEGTY GVVYK (the substrate for BTK and c-SRC);
0.33 mg/ml myelin basic protein (the substrate for VEGFR2);
50 μM EAIYAAPFAKKK (the substrate for FLT3);
10 mM a solution of magnesium acetate and γ-33 P-ATP;
Terminating buffer solution (3% buffer solution of phosphate salt);
Washing buffer solution (75 mM phosphate solution);
Methanol;
Filtermat A membrane;
EGFRs (including wild type, L858R mono mutant type and L858R/T790M double mutant type EGFRs), VEGFR2, ALK, BTK, c-KIT, c-SRC, MET, PDGFRα, FLT3 kinase, and the tested compounds.
2. Procedure
To a reaction tube were successively added the buffer solution (8 mM MOPS, pH 7.0, 0.2 mM EDTA, 10 mM MnCl2), the kinase to be tested (5-10 mU) (EGFR/ALK/BTK/c-KIT/c-SRC/MET/PDGFRα/VEGFR2), the substrate for the kinase to be tested (a reference material), 10 mM of the solution of magnesium acetate and γ-33 P-ATP, and different concentrations of the tested compounds. The reaction was started by adding MgATP (the final concentration of ATP is the Km value of the corresponding kinase, i.e., 10 μM for EGFRWild Type, 200 μM for EGFR L858R, 45 μM for EGFR L858R/T790M, 200 μM for ALK, 200 μM for BTK, 200 μM for c-KIT, 90 μM for VEGFR2, 200 μM for c-SRC, 45 μM for MET, 120 μM for PDGFRα, and 200 μM for FLT3), and incubated at room temperature for 40 minutes. The reaction was terminated with 54 of the 3% phosphate buffer solution. 10 μL of the reaction liquid was titrated on the Filtermat A membrane. The membrane was washed with 75 mM of the phosphate solution thrice (5 minutes each time), and then with methanol once, and finally dried. The membrane was subjected to a scintillation counting. The value of the scintillation counting reflected the phosphorylation level of the substrate and therefore could characterize the inhibition of the kinase activity.
3. Results
Through the above procedures, the inhibitory activities of the present compounds were tested for the kinases EGFRs (comprising wild type, L858R mutant type and L858R/T790M double mutant type), VEGFR2, ALK, BTK, c-KIT, c-SRC, MET, PDGFRα, and FLT3. The kinase inhibitory activities (IC50 values) of the tested compounds for EGFRs (comprising wild type, L858R mutant type and L858R/T790M double mutant type) and VEGFR2 are shown in Table 1. The activity inhibition ratio (%) of the tested compounds at 10 μM for the kinases ALK, BTK, c-KIT, c-SRC, MET, PDGFRα, and FLT3 are shown in Table 2.
The results indicated that the tested compounds had strong inhibitory activities on the wild type, L858R mutant type and L858R/T790M double mutant type EGFRs, and some of the tested compounds also had good inhibitory activities on VEGFR2, ALK, BTK, c-KIT, c-SRC, MET, PDGFRα, FLT3 kinases.










TABLE 1









EGFR(IC50, μM)












Tested
Wild

L858R/



Compounds
Type
L858R
T790M
VEGFR2(IC50, μM)














8-1
0.003
0.008
0.090
>1.000


8-2
0.002
0.001
<0.001
>1.000


8-3
0.025
0.060
0.245
>1.000


8-4
0.030
0.018
0.046
>1.000


8-5
0.004
0.005
0.006
>1.000


8-6
0.001
0.0004
0.018
>1.000


8-7
0.007
0.001
0.128
>1.000


8-9
<0.001
0.0004
0.006
>1.000


8-10
0.005
0.005
0.046
>1.000


8-11
0.007
0.004
0.005
0.140


8-12
0.004
0.003
0.003
0.050


8-13
0.004
0.003
0.009
>1.000


8-15
0.003
0.001
0.015
0.004


8-16
0.003
0.001
0.012
>1.000


8-17
0.004
0.001
0.004
>1.000


8-18
0.030
0.005
0.057
0.021


8-19
0.005
0.004
0.004
>1.000


8-21
0.011
0.005
0.005
>1.000


8-22
0.117
0.010
0.087
>1.000


8-23
0.007
0.002
0.041
>1.000


8-24
0.012
0.003
0.522
>1.000


8-25
0.029
0.005
0.458
>1.000


8-26
<0.001
0.0004
<0.001
0.007


8-27
0.034
0.005
0.054
0.009


8-29
0.031
0.004
0.064
>1.000


8-30
0.015
0.005
0.228
>1.000


8-38
0.002
0.0006
0.037
>1.000

















TABLE 2







Tested
The kinase activity inhibitory rate (%) by the tested


Com-
compounds at 10 μM














pounds
ALK
BTK
c-KIT
c-SRC
MET
PDGFRα
FLT3

















8-1 
23
46
26
11
83
43
85


8-2 
63
98
90
99
100
84
78


8-3 
16
11
54
67
2
20
0


8-4 
64
45
23
12
67
16
43


8-5 
54
78
54
32
64
19
35


8-6 
15
46
21
82
14
39
56


8-7 
61
12
8
28
0
9
0


8-9 
36
88
96
98
67
88
76


8-10
45
32
3
23
64
87
87


8-11
33
77
67
98
93
58
67


8-12
80
78
82
98
93
55
56


8-13
78
45
87
31
100
90
89


8-15
40
95
94
97
45
94
90


8-16
17
57
62
90
93
39
45


8-17
35
100
89
99
52
72
86


8-18
13
56
76
93
7
60
75


8-19
90
71
12
97
97
34
43


8-21
97
92
101
98
99
100
78


8-22
70
63
10
98
69
43
87


8-23
3
30
26
83
1
28
68


8-24
8
14
48
75
0
29
75


8-25
13
45
8
78
0
21
32


8-26
71
99
94
99
86
95
86


8-27
30
74
93
98
36
68
83


8-29
15
60
36
97
50
42
99


8-30
8
25
0
67
0
21
43


8-38
22
93
63
93
74
55
86









Assay 2: The Test of the Arylamino Purine Derivatives for the In-Vitro Tumor Cell Proliferation Inhibition

The object of this assay was to test the inventive compounds for the inhibitory activity of in-vitro tumor cell proliferation. The MTT (tetreamethyl-azo-zole-salt) colorimetric method was used in this assay.
1. Materials
1.1 Main Agents
RPMI-1640, fetal bovine serum, pancreatin and the like were purchased from Gibco BRL Company (Invitrogen Corporation, USA). The IMDM culture medium was purchased from ATCC (American Type Culture Collection). Tetreamethyl-azo-zole-salt (MTT) and dimethylsulfoxide (DMSO) were the products available from Sigma Company (USA). The arylamino purine derivatives were synthesized by the present inventors. In this in-vitro assay, 100% DMSO was formulated into a 10 mM stocking solution and preserved in a freezer at −20° C. and in dark place for use. The stocking solution was diluted with a complete culture solution to a desired concentration immediately before use.
1.2 Cell Lines and their Culturing
Human non-small cell lung carcinoma cell strains HCC827, PC-9, H1975 (EGFR L858R/T790M mutation) and H292 (EGFR WT), and other tumor type cell strains, including human acute myelogenous leukemia cell strain MV4-11, human chronic granulocytic leukemia cell strain K562, human squamous cell carcinoma cell strain A431, human breast carcinoma cell strains MDA-MB-468 and BT 474, human colon cancer cell strains SW480, HCT116 and SW620, human liver cancer cell strain Hep G2, human gastric cancer cell strain MK-45, and human malignant melanoma cell strain A375 used in this assay were all purchased from ATCC company, USA and kept in the laboratory. All of the above-mentioned non-small cell lung carcinoma cell strains and the breast carcinoma cell strain BT 474 were cultured with a RPMI-1640 complete culture medium containing 10% fetal bovine serum, 100 U/ml penicillin and 100 μg/ml streptomycin under 5% CO2 and at 37° C. The other cell strains were cultured with a DMEM complete culture medium containing 10% fetal bovine serum (having a MV4-11 cell content of 20%), 100 U/ml penicillin and 100 μg/ml streptomycin under 5% CO2 and 37° C.
2. Procedure
A cell suspension having a cell concentration of 1-2×104 cells/ml was treated with a complete cell culture solution to adjust the cell concentration, wherein the cell concentrations for HCC827 and MV4-11 were adjusted to 6×104 cells/ml and 1×105 cells/ml respectively. The cell suspension was inoculated in a 96-well plate with 200 μl cell suspension/well and cultured overnight. Next day, the supernatant was drawn off and discarded. Then, the cells were treated with the tested compounds in a gradient concentration respectively. In the meanwhile, a negative control group free of the drug substance and an isovolumetric solvent control group (having a DMSO concentration of 1%) were used. The triplicate wells were used for each of dose groups. The culturing was conducted at 37° C. under 5% CO2. After 72 hours, 200 MTT agent having a concentration of 5 mg/ml was added to each of wells. The culturing was further conducted for 2-4 hours. The supernatant was discarded. Then 1504 DMSO was added to each of wells. The contents in the well were mixed homogenously by oscillation for 15 minutes. The absorbance (A) value was measured with a microplate reader at λ=570 nm (the A value is in the direct proportion to the number of living cells) and averaged. The relative cell proliferation inhibitory rate is (A570 control-group−A570 dose-group)/A570 control-group×100%. The assay was repeated for at least three times. The data was expressed as number average. The statistical data were analyzed using t-test. P<0.05 was considered significant. The cell proliferation inhibition of the following compounds was expressed as IC50 or inhibitory rate.
3. Results
According to the above-mentioned procedure, human non-small cell lung carcinoma cell strains HCC827, PC-9 (EGFR delE746-A750 deletion mutation), H1975 (EGFR L858R/T790M mutation) and H292 (EGFR WT), and other tumor type cell strains, including human acute myelogenous leukemia cell strain MV4-11, human chronic granulocytic leukemia cell strain K562, human squamous cell carcinoma cell strain A431, human breast carcinoma cell strains MDA-MB-468 and BT 474, human colon cancer cell strains SW480, HCT116 and SW620, human liver cancer cell strain Hep G2, human gastric cancer cell strain MK-45, and human malignant melanoma cell strain A375 were subjected to the proliferation inhibition activity test.
The proliferation inhibitory activities (IC50) of the tested compounds for human non-small cell lung carcinoma cell strains HCC827, PC-9 and H1975 are shown in the table 3. The proliferation inhibitory activities (IC50) of the tested compounds for human tumor cell strains MV4-11, K562, A431, MDA-MB-468, BT474, SW480, HCT116, HepG2, SW620, MK-45, H292 and A375 are shown in the tables 4 and 5. The results indicated that the tested compounds had strong inhibitory activities on the cell strains HCC827 and PC-9 which were sensitive to Gefitinib; some of the tested compounds also had a good inhibitory activity on the cell strain H1975 which was resistant to Gefitinib; and in addition, some of the tested compounds also had good inhibitory activities on the other tumor cell strains including human MV4-11, K562, A431, MDA-MB-468, BT474, SW480, HCT116, HepG2, SW620, MK-45, H292, A375 and the like.












TABLE 3







Tested
HCC827
PC-9
H1975



Compounds
(IC50, μM)
(IC50, μM)
(IC50, μM)





















8-1 
0.0020
0.1047
7.6



8-2 
0.0013
0.5580
>10



8-3 
>1
>1
>10



8-4 
0.0030
0.0680
0.86



8-5 
0.0013
0.5580
>10



8-6 
0.0002
0.0137
>10



8-7 
0.0075
0.0121
>10



8-9 
0.0042
0.0723
>10



8-10
0.0001
0.0009
2



8-11
0.0002
0.0002
1.726



8-12
0.0002
0.0002
1.929



8-13
0.0002
0.0003
2



8-15
0.0002
0.0064
>10



8-16
0.0002
0.0168
1.822



8-17
0.0002
0.0025
1.904



8-18
0.1172
0.0349
>10



8-19
0.9116
0.4591
7.32



8-21
0.0516
0.0859
7.312



8-22
>1
2.7947
>10



8-23
0.0002
0.0359
>10



8-24
0.0002
0.0060
>10



8-25
0.0528
0.2582
>10



8-26
0.0011
0.0002
0.369



8-27
0.0823
0.1149
>10



8-29
0.0072
0.0100
>10



8-30
0.4828
0.0551
>10



8-34
0.0631
0.4199
6.3076



8-35
0.0002
0.0015
1.5162



8-36
0.0002
0.0209
0.8518



8-37
0.1297
0.3476
>10



8-38
0.0005
0.0007
0.6445



8-41
0.0165
0.2092
0.5013



8-42
4.2614
3.3210
>10



8-43
0.0003
0.0519
0.2309



8-44
0.0003
0.0015
0.6947



8-45
0.0002
0.0097
1.1359



8-46
0.0003
0.0035
0.7702



8-47
0.0003
0.0163
0.8844



8-48
0.0002
0.0262
4.5828



8-49
0.0003
0.0002
0.3103



8-50
0.0003
0.0035
0.1822



8-51
0.0002
0.0095
0.6302



8-52
0.0003
0.0006
0.2714



8-53
0.0002
0.0002
0.5516



8-54
0.0063
0.2045
1.7011



8-55
0.0002
0.0010
0.3228



8-56
0.0006
0.0070
>10



8-57
0.0280
0.1773
>10



8-58
0.0002
0.0181
0.2521



8-59
0.0002
0.0021
0.2482



8-60
0.0002
0.0117
0.4731



8-61
0.0143
0.1165
4.2103



8-62
0.0002
0.0091
0.3920























TABLE 4





Tested
MV4-11
K562
A431
MDA-MB-468
BT474
SW480


Compounds
(IC50, μM)
(IC50, μM)
(IC50, μM)
(IC50, μM)
(IC50, μM)
(IC50, μM)





















8-1 
0.089
1.078
2.059
2.059
2.132
5.725


8-2 
1.768
3.734
9.293
6.134
1.859
>10


8-3 
>10
>10
12.9
>10
>10
>10


8-4 
9.772
1.391
1.986
4.766
0.635
>10


8-5 
1.768
3.734
9.293
6.134
1.859
>10


8-6 
2.845
7.354
2.388
2.985
0.597
>10


8-7 
>10
>10
>10
>10
5.818
>10


8-9 
0.235
0.143
1.527
1.321
1.032
>10


8-10
0.102
0.106
1.582
0.7
0.226
>10


8-11
3.755
0.366
7.671
1.189
0.575
>10


8-12
1.711
0.221
2.572
3.215
2.143
>10


8-13
0.512
0.086
1.293
2.101
0.202
>10


8-15
0.197
0.303
9.397
2.301
1.918
>10


8-16
4.799
2.523
5.876
1.959
1.371
6.481


8-17
0.684
0.003
8.464
0.783
0.635
>10


8-18
0.183
1.221
6.348
3.174
2.116
>10


8-19
4.778
2.791
9.316
9.981
4.991
>10


8-21
0.279
1.335
5.722
0.89
1.589
>10


8-22
0.283
0.921
5.52
1.725
1.725
>10


8-23
0.241
5.198
7.037
5.221
6.81
>10


8-24
0.288
1.422
9.65
9.65
2.412
>10


8-25
7.896
>10
>10
>10
9.988
>10


8-26
0.093
0.12
1.046
0.299
0.107
3.438


8-27
0.088
0.777
2.35
1.959
1.175
2.597


8-29
0.004
0.24
9.018
8.116
2.255
>10


8-30
7.962
5.756
>10
>10
4.458
>10


8-34
0.1248
1.5971
2.5734
3.2589
2.6282
9.9542


8-35
0.1353
0.2322
0.6351
1.9193
0.1919
5.9306


8-36
0.0010
0.0760
0.4630
>10
0.6389
8.6758


8-37
0.1016
0.8882
>10
>10
4.5220
>10


8-38
0.7297
0.2685
0.1790
0.2805
1.6809
>10


8-41
1.9710
0.1460
2.7386
4.4875
2.0921
5.3955


8-42
6.6387
0.9223
2.2395
1.7005
5.1014
>10


8-43
0.0212
0.1225
0.4844
1.0296
1.8265
5.7535


8-44
1.1377
0.0965
0.4089
2.6656
0.6815
6.1586


8-45
0.0594
0.2184
1.8855
2.8739
0.8702
7.7514


8-46
4.6852
0.2505
0.4263
0.7511
0.7917
4.0051


8-47
0.2242
0.0972
0.2989
3.1902
0.5542
5.5316


8-48
0.5831
0.2300
0.7879
2.1296
0.5254
9.8918


8-49
0.1930
0.1207
0.5865
0.9871
2.6675
5.0433


8-50
0.0644
0.0305
0.0829
0.2072
2.1632
2.7164


8-51
0.0036
0.0113
0.2476
0.3508
0.7635
6.7231


8-52
0.6419
0.1478
0.0796
0.7367
0.1653
4.6233


8-53
0.8986
0.0953
0.2486
0.6216
1.5126
4.2104


8-54
1.0737
0.8627
2.0614
3.3489
3.9647
6.7257


8-55
0.6823
0.0880
0.1982
2.1800
0.4558
5.3193


8-56
4.4271
>10
>10
>10
>10
>10


8-57
6.1616
0.3521
5.7699
>10
>10
9.7634


8-58
0.1806
0.0671
0.2416
0.7450
0.0805
5.5452


8-59
0.0998
0.1113
0.4973
6.8377
1.1017
4.7470


8-60
0.1459
0.4510
2.0465
7.1257
1.0240
4.2550


8-61
1.6785
1.0075
5.2634
6.1396
1.8568
8.1211


8-62
0.5772
0.1391
0.4932
0.5553
0.1867
3.4996






















TABLE 5





Tested
HCT116
HEPG2
SW620
MK45
H292
A375


Compounds
(IC50, μM)
(IC50, μM)
(IC50, μM)
(IC50, μM)
(IC50, μM)
(IC50, μM)





















8-1 
>10
>10
>10
6.7960
1.3880
3.0211


8-2 
7.812
>10
5.567
>1
0.8829
1.5864


8-3 
>10
>10
>10
>10
1.3039
>10


8-4 
>1
>10
>1
2.1844
0.1489
>10


8-5 
7.812
>10
5.567
>1
>1
>1


8-6 
3.295
2.571
2.135
0.5398
0.1273
6.9321


8-7 
>10
>10
>10
3.7625
0.1377
1.3124


8-9 
0.518
>10
0.524
0.7635
0.1596
0.5861


8-10
1.091
4.797
6.056
>1
0.0105
1.4891


8-11
1.682
11.215
1.96
>1
0.0088
1.3865


8-12
4.008
6.606
1.292
>1
0.0107
3.0927


8-13
2.543
5.962
3.675
>1
0.0061
0.8384


8-15
2.2
7.794
2.992
0.7095
0.2205
0.7747


8-16
0.766
10.445
0.864
>1
0.1293
3.1005


8-17
0.813
>10
>1
0.6642
0.0296
0.1896


8-18
6.289
>10
4.003
1.5206
0.5311
6.4623


8-19
>1
5.42
1.993
1.0055
0.3161
1.6636


8-21
0.772
8.933
2.225
1.3279
1.1603
0.7248


8-22
1.829
>10
1.922
0.5108
1.7251
1.2973


8-23
>10
>10
>10
2.7898
0.1544
3.1756


8-24
1.382
>10
>10
8.0818
0.1713
2.4004


8-25
>10
>10
>10
5.3511
1.6455
>10


8-26
0.239
6.255
0.282
>1
0.0107
0.3777


8-27
>1
>1
>1
6.1109
4.1856
9.7932


8-29
2.48
>10
5.796
8.1682
0.1804
1.8758


8-30
>10
>10
4.217
5.4680
0.2006
>10


8-34
10.4821
8.4627
>10
1.6645
1.2331
6.5704


8-35
0.6756
4.3356
2.3031
1.1145
0.0203
3.2455


8-36
1.1223
3.5159
3.6139
1.5518
0.0341
1.4481


8-37
>10
>10
>10
>10
5.8524
>10


8-38
0.4426
2.9046
0.3700
1.7897
0.0073
>10


8-41
5.4311
2.1988
2.1402
3.3432
0.2678
>10


8-42
>10
>10
>10
5.1020
0.6734
0.4982


8-43
2.5663
5.9270
5.7535
0.2338
0.1017
1.9069


8-44
>10
>10
2.5098
1.3528
0.0029
1.7417


8-45
7.0842
>10
4.9666
3.7400
0.0139
9.2557


8-46
9.0658
>10
3.6194
0.4170
>10
0.3999


8-47
6.0851
8.4636
4.9480
1.1820
0.0442
1.5739


8-48
>10
>10
2.9367
0.2969
0.0185
2.0848


8-49
1.4365
9.2439
1.4365
0.5426
0.0045
1.9811


8-50
2.4678
8.2487
0.9635
0.2176
>10
1.5602


8-51
4.1374
8.2150
0.7326
0.2476
0.0223
0.5386


8-52
5.9255
7.2296
1.5690
0.4566
0.0040
0.9736


8-53
>10
>10
2.1135
0.5594
0.0041
8.0705


8-54
6.9601
5.9796
6.6383
0.6884
0.2791
5.5815


8-55
6.5500
4.6772
2.9748
1.7216
>10
2.7686


8-56
>10
>10
>10
>10
0.0810
>10


8-57
7.8646
>10
8.3682
4.5862
0.7518
>10


8-58
7.4640
>10
1.2504
0.5638
0.0040
0.6604


8-59
5.7188
>10
1.3344
0.8081
0.0033
1.1976


8-60
6.4206
>10
1.2521
1.8443
0.0545
9.3025


8-61
>10
>10
5.9796
2.4503
0.1879
>10


8-62
3.5197
6.8330
1.3023
0.4371
0.0055
1.0210









Assay 3: The In-Vivo Anti-Tumor Test for Compound 8-10

The object of this assay was to determine the in-vivo anti-tumor effect of the present compound. In this assay, a nude mouse subcutaneously transplanted non-small cell lung carcinoma model was used to test the present compound 8-10 for the in-vivo anti-tumor activity. The used cell strain was human non-small cell lung carcinoma cell strain HCC827.
1. Materials
RPMI-1640, fetal bovine serum, pancreatin and the like were purchased from Gibco BRL company (Invitrogen Corporation, USA); RPMI 1640 culture medium was purchased from ATCC (American Type Culture Collection); human non-small cell lung carcinoma cell strain HCC827 was purchased from ATCC company, USA; and BALB/C nude mice were purchased from Institute of Zoology, Chinese Academy of Sciences.
2. Procedure
BALB/C nude mice, aged 6-8 weeks, were inoculated with HCC827 cell subcutaneously at the posterior segment of rib in a concentration of about 5×106 cells/0.1 ml per mouse. Upon the growth of the tumor up to 200-300 mm3 (about 20 days), mice were grouped (n=6) and administrated intragastrically.
Groups:
Solvent control group: (5% DMSO+1% TWEEN80+94% water);
Gefitinib group: 100 mg/kg q.d.;
Compound 8-10 group: 2 mg/kg q.d.;
Compound 8-10 group: 5 mg/kg q.d.;
Compound 8-10 group: 10 mg/kg q.d.;
Compound 8-10 group: 20 mg/kg q.d.
(Each of drug groups was dissolved in 5% DMSO+1% TWEEN80+94% water).
Observation Indices: the mice were measured every three days for the weight, and the length and width of tumor, and the tumor volume was calculated as length×width2×0.52. The mice were observed for the reactions such as diarrhea, convulsion, exanthema, and substantial weight reduction.

3. Results

The measured tumor growth curves for each of the groups are shown in FIG. 1. The photograph of tumors obtained by dissection after the assay is shown in FIG. 2. The results indicated that the tested compound 8-10 had a substantial in-vivo growth inhibition for the EGFRdelE746-A750 mutated human non-small cell lung carcinoma cell strain HCC827. Upon administrating in 2 mg/kg q.d. or higher, the tumor growth could be substantially inhibited, or even the tumor could be eliminated. In the course of administration, the nude mice did not show the untoward reactions such as weight reduction, exanthema, and diarrhea, which indicated that under the test doses, the tested compound 8-10 had a low toxicity in the dose range of administration.
Assay 4: The In-Vivo Anti-Tumor Test for Compound 8-29
The object of this assay was to determine the in-vivo anti-tumor effect of the present compound. In this assay, a NOD-SCID mouse subcutaneously transplanted human leukemic solid tumor model was used to test the present compound 8-29 for the in-vivo anti-tumor activity. The used cell strain was human acute myelogenous leukemia cell strain MV4-11.
1. Materials
IMDM, fetal bovine serum, pancreatin and the like were purchased from Gibco BRL company (Invitrogen Corporation, USA); IMDM culture medium was purchased from ATCC (American Type Culture Collection), human leukemia cell strain MV4-11 was purchased from ATCC company, USA; and the NOD-SCID mice were purchased from Laboratory Animal Center, Peking Union Medical College, China.
2. Procedure
NOD-SCID mice, aged 6-8 weeks, were inoculated with MV4-11 cell subcutaneously at the posterior segment of rib in a concentration of about 1×107 cells/0.1 ml per mouse. Upon the growth of the tumor up to 400-500 mm3 (about 20 days), mice were grouped (n=6) and administrated intragastrically.
Groups:
Solvent control group: (5% DMSO+25% PEG400+70% water);
Compound 8-29 group: 5 mg/kg q.d.;
Compound 8-29 group: 10 mg/kg q.d.;
Compound 8-29 group: 20 mg/kg q.d.
(Each of drug groups was dissolved in 5% DMSO+25% PEG400+70% water)
Observation Indices: the mice were measured every three days for the weight, and the length and width of tumor, and the tumor volume was calculated as length×width2×0.52. The mice were observed for the reactions such as diarrhea, convulsion, exanthema, and substantial weight reduction.

3. Results

The measured tumor growth curves for each of the groups are shown in FIG. 3. The results indicated that the tested compound 8-29 had a substantial in-vivo growth inhibition for the FLT3-ITD mutated human acute myelogenous leukemia cell strain MV4-11. Upon administrating in 5 mg/kg q.d. or higher, the tumor growth could be substantially inhibited, or even the tumor could be eliminated. In the course of administration, the mice did not show the untoward reactions such as weight reduction, exanthema, and diarrhea, which indicated that under the test doses, the tested compound 8-29 had a low toxicity in the dose range of administration.

## Claims

1. An arylamino purine derivative represented by formula I:




wherein
R1 is selected from the group consisting of —H, —CmH(2m+1), C3-C7 cycloalkyl, —CmH(2m+1) substituted by C3-C7 cycloalkyl, and C3-C7 cycloalkyl substituted by —CmH(2m+1);
R2 is selected from the group consisting of —H, —NH2, —OH, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1), and —NHCmH(2m+1);
R3 is selected from the group consisting of





and C3-C7 cycloalkyl;
R4 is selected from the group consisting of




R5-R8 are each independently selected from the group consisting of —H, —F, —Cl, —Br, —CF3, and —OCmH(2m+1);
R9-R13 are each independently selected from the group consisting of —H, —F, —Cl, —Br, —CF3, —CmH(2m+1), —OCmH(2m+1),




m=1-8; and
n=0-4.

2. The arylamino purine derivative of claim 1, wherein R1 is selected from the group consisting of —H, —CmH(2m+1), C3-C7 cycloalkyl, and —CmH(2m+1) substituted by C3-C7 cycloalkyl.

3. The arylamino purine derivative of claim 1, wherein R2 is selected from the group consisting of —H, —CmH(2m+1), —OCmH(2m+1), and —NHCmH(2m+1).

4. The arylamino purine derivative of claim 1, wherein
R4 is selected from the group consisting of

5. The arylamino purine derivative of claim 1, wherein R4 is

6. The arylamino purine derivative of claim 1, wherein R1 is selected from the group consisting of —H, —CmH(2m+1),

7. The arylamino purine derivative of claim 1, wherein the arylamino purine derivative is selected from the group consisting of:

8. A method of treating tumors, comprising administering an effective amount of the arylamino purine derivative according to claim 1 to a subject in need thereof, wherein the tumors are selected from the group consisting of human non-small cell lung carcinoma, human acute myelogenous leukemia, human chronic granulocytic leukemia, human squamous cell carcinoma, human breast carcinoma, human colon cancer, human liver cancer, human gastric cancer, and human malignant melanoma.

9. A pharmaceutical composition comprising the arylamino purine derivative according to claim 1, or its pharmaceutical acceptable salt.

10. An arylamino purine derivative selected from the group consisting of:

11. A method of treating tumors, comprising administering an effective amount of the arylamino purine derivative according to claim 10 to a subject in need thereof, wherein the tumors are selected from the group consisting of human non-small cell lung carcinoma, human acute myelogenous leukemia, human chronic granulocytic leukemia, human squamous cell carcinoma, human breast carcinoma, human colon cancer, human liver cancer, human gastric cancer, and human malignant melanoma.

12. A pharmaceutical composition comprising the arylamino purine derivative according to claim 10, or its pharmaceutical acceptable salt.