Multi-ring kinase inhibitor

Abstract

The present invention belongs to the field of pharmaceutical technology, and particularly relates to polycyclic DNA-PK kinase inhibitor compounds represented by general formula (I), pharmaceutically acceptable salts thereof or isomers thereof, pharmaceutical compositions and preparations containing the compounds, pharmaceutically acceptable salts thereof or isomers thereof, methods for preparing the compounds, pharmaceutically acceptable salts thereof or isomers thereof, and uses of the compounds, pharmaceutically acceptable salts thereof or isomers thereof.

Description

Technical Field

[0001] The present invention belongs to the technical field of medicine, and particularly relates to polycyclic DNA-PK kinase inhibitor compounds, pharmaceutically acceptable salts or isomers thereof, pharmaceutical compositions and preparations containing the compounds, pharmaceutically acceptable salts or isomers thereof, methods for preparing the compounds, pharmaceutically acceptable salts or isomers thereof, and uses of the compounds, pharmaceutically acceptable salts or isomers thereof. Background Art

[0002] Cancer is a malignant disease that is difficult to treat worldwide, with great treatment difficulty and high mortality rate, bringing a heavy burden to patients and their families, and is a major disease affecting the health of residents in our country. In recent years, the incidence of cancer in our country has increased significantly, and its mortality rate has also shown a gradually increasing trend, and cancer prevention and treatment are facing a severe situation.

[0003] Currently, radiotherapy and chemotherapy are the most effective means for treating cancer in addition to surgical resection. At the same time, radiotherapy is the most effective non-surgical treatment for malignant tumors. Radiation and a considerable number of anticancer drugs can directly or indirectly act on DNA or the DNA metabolism process, thereby causing DNA damage. Among them, DNA double strand break (DSB) is the most lethal to cancer cells. After DNA damage, a series of cellular responses such as damaged DNA repair will be triggered, and the result of the repair is to increase the survival of cancer cells, which is also one of the mechanisms of tumor cell resistance to radiotherapy and chemotherapy. If DNA double strand breaks are not repaired in a timely and complete manner, cancer cells will die due to apoptosis or / and mitotic disorders. Therefore, as long as the repair of these DNA damages is inhibited, the sensitivity of cancer cells to radiotherapy and chemotherapy can be increased, and cell proliferation can be inhibited.

[0004] In higher eukaryotic cells such as human cells, the repair of DSB is mainly carried out through DNA non-homologous end joining (NHEJ) dominated by DNA-dependent protein kinase (DNA-PK), thereby repairing the damaged DNA and maintaining cell viability and genomic stability. NHEJ repair mainly participates in DNA damage repair in the G1 / S phase and does not require a DNA end ligation template. NHEJ repair requires the coordinated cooperation of many proteins and signaling pathways. The heterodimer of the Ku70 / 80 subunits and the catalytic subunit DNA-dependent protein kinase (DNA-PKcs) together form an active DNA-PK enzyme complex.

[0005] DNA-PKcs belongs to the phosphatidylinositol 3-kinase (PI3K) superfamily and is a serine / threonine protein kinase; the PI3K superfamily also includes ATM, ATR, mTOR, and four PI3K isoforms. The kinase activity of DNA-PK can only be activated when it binds to broken DNA. The important function of Ku is to bind to the DNA ends and recruit DNA-PKcs, and the two form the DNA-PK holoenzyme and activate DNA-PKcs; the activated DNA-PKcs guides the Artemis protein (an endonuclease) to bind to the damaged site, and relies on its ribozyme activity to process the DNA ends for ligation and repair. Then, the XRCC4 / DNA ligase IV complex is recruited by the activated DNA-PKcs, and finally, DNA ligase IV locates and ligates the ends of the broken DNA double strands to complete the repair. XRCC4 is a protein that forms a complex with DNA ligase IV and can increase the activity of DNA ligase IV. There are 40 amino acid residues in DNA-PKcs that can be autophosphorylated, and the most typical autophosphorylation sites occur at Ser2056 (POR cluster) and Thr2609 (ABCDE cluster). NHEJ is considered to proceed through three key steps: recognition of DSB - Ku70 / 80 binds to the incomplete DNA ends, and two molecules of DNA-PKcs are recruited to the adjacent sides of the DSB; DNA processing is carried out to remove non-ligatable ends or other damaged forms at the ends; finally, the DNA ends are ligated.

[0006] Since tumor cells have a relatively high basal level of endogenous replication stress (oncogene-induced replication stress) and DNA damage, and the DNA repair mechanism is less efficient in tumor cells, tumor cells are more sensitive to DNA-PK.

[0007] Currently, the development of highly efficient and selective DNA-PK inhibitors has important clinical significance. They can synergistically enhance the effects of radiotherapy and chemotherapy, effectively inhibit tumor growth, and at the same time can effectively reduce the damage to normal cells and reduce side effects. Summary of the Invention

[0008] The technical problem to be solved by the present invention is to provide a polycyclic compound with a novel structure and good inhibitory effect on DNA-PK. Further, such compounds can be used to increase the sensitivity of a subject to radiotherapy and / or one or more anticancer agents. Furthermore, such compounds can be used in combination with radiotherapy and / or one or more anticancer agents for the prevention and / or treatment of benign tumors or cancers.

[0009] The technical solution of the present invention is as follows:

[0010] In one aspect, the present invention provides a compound represented by the following general formula (I), its pharmaceutically acceptable salt, or its isomer,

[0011]

[0012] Wherein,

[0013] X1, X2, X3, and X4 are each independently selected from C(R 4 ) or N;

[0014] X5 and X6 are each independently selected from CH(R 5 ), C(R 6 ), N(R 7 ) or N;

[0015] X is selected from CH2, NH, O or S;

[0016] R 1 is selected from H, halogen, hydroxyl, amino, nitro, cyano, C 1-6 alkyl, C 1-6 alkylamino, di(C 1-6 alkyl)amino, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, amino C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, halo C 1-6 alkoxy, halo C 1-6 alkylthio, hydroxy C 1-6 alkoxy, hydroxy C 1-6 alkylthio, amino C 1-6 alkoxy, amino C 1-6 alkylthio;

[0017] R 2 , R 3 are each independently selected from H, halogen, hydroxyl, amino, nitro, cyano, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, amino C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 alkoxy, amino C 1-6 alkoxy, an optionally 1-3 Q1-substituted 3-8 membered cycloalkyl or 3-8 membered heterocyclic group; each Q1 is independently selected from halogen, hydroxyl, amino, nitro, cyano, C 1-6 alkyl, halo C 1-6 alkyl, C 1-6 alkoxy or halo C 1-6 alkoxy;

[0018] Or R 2 , R3 and together with the carbon atom to which it is attached form a 3-8 membered cycloalkyl or 3-8 membered heterocyclic group optionally substituted by 1-3 Q2s; each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, C 1-6 alkyl, halo C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6 alkoxy or halo C 1-6 alkoxy;

[0019] each R 4 、each R 5 、R 6 is independently selected from H, halogen, hydroxy, amino, nitro, cyano, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, amino C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 alkoxy or amino C 1-6 alkoxy;

[0020] each R 7 is independently selected from H, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl or amino C 1-6 alkyl;

[0021] the dashed bond is a chemical bond or does not exist, and adjacent dashed bonds are not both chemical bonds.

[0022] In certain embodiments, the compound of formula (I), its pharmaceutically acceptable salt or its isomer, further has the structure shown in the following formula (IIa),

[0023]

[0024] wherein X5 is selected from CH(R 5 ) or N(R 7 );

[0025] X6 is selected from C(R 6 ) or N;

[0026] X is selected from CH2, NH, O or S;

[0027] R 1 is selected from H, halogen, hydroxy, amino, nitro, cyano, C 1-6 alkyl, halo C1-6 alkyl, hydroxy C 1-6 alkyl, amino C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 alkoxy or amino C 1-6 alkoxy;

[0028] R 2 、R 3 are each independently selected from H, halogen, hydroxy, amino, nitro, cyano, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, amino C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 alkoxy, amino C 1-6 alkoxy, a 3- to 8-membered cycloalkyl or 3- to 8-membered heterocyclic group optionally substituted by 1 or 2 Q1s; each Q1 is independently selected from halogen, hydroxy, amino, nitro, cyano, C 1-6 alkyl, halo C 1-6 alkyl, C 1-6 alkoxy or halo C 1-6 alkoxy;

[0029] or R 2 、R 3 together with the carbon atom to which it is attached form a 3- to 8-membered cycloalkyl or 3- to 8-membered heterocyclic group optionally substituted by 1 or 2 Q2s; each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, C 1-6 alkyl, halo C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6 alkoxy or halo C 1-6 alkoxy;

[0030] R 5 、R 6 are each independently selected from H, halogen, hydroxy, amino, nitro, cyano, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, amino C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 alkoxy or amino C 1-6 alkoxy;

[0031] R 7Selected from H, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl or amino C 1-6 alkyl.

[0032] In certain embodiments, the compound of formula (I), its pharmaceutically acceptable salt or its isomer, further has the structure shown in the following general formula (IIb),

[0033]

[0034] wherein X5 is selected from C(R 6 ) or N;

[0035] X6 is selected from CH(R 5 ) or N(R 7 );

[0036] X is selected from CH2, NH, O or S;

[0037] R 1 is selected from H, halogen, hydroxy, amino, nitro, cyano, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, amino C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 alkoxy or amino C 1-6 alkoxy;

[0038] R 2 , R 3 are each independently selected from H, halogen, hydroxy, amino, nitro, cyano, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, amino C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 alkoxy, amino C 1-6 alkoxy, 3-8 membered cycloalkyl or 3-8 membered heterocyclic group optionally substituted by 1-2 Q1s; each Q1 is independently selected from halogen, hydroxy, amino, nitro, cyano, C 1-6 alkyl, halo C 1-6 alkyl, C 1-6 alkoxy or halo C 1-6 alkoxy;

[0039] Or R 2 , R 3and, together with the carbon atom to which it is attached, form a 3-8 membered cycloalkyl or 3-8 membered heterocyclic group optionally substituted by 1-2 Q2; each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, C 1-6 alkyl, halo C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6 alkoxy or halo C 1-6 alkoxy;

[0040] R 5 、R 6 are each independently selected from H, halogen, hydroxy, amino, nitro, cyano, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, amino C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 alkoxy or amino C 1-6 alkoxy;

[0041] R 7 is selected from H, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl or amino C 1-6 alkyl.

[0042] In certain embodiments, R 2 、R 3 are each independently selected from H, halogen, hydroxy, amino, nitro, cyano, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, amino C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 alkoxy, amino C 1-6 alkoxy, a 3-8 membered cycloalkyl or 3-8 membered heterocyclic group optionally substituted by 1-2 Q1; each Q1 is independently selected from halogen, hydroxy, amino, nitro, cyano, C 1-6 alkyl, halo C 1-6 alkyl, C 1-6 alkoxy or halo C 1-6 alkoxy.

[0043] In certain embodiments, R 2 、R 3 are each independently selected from H, halogen, hydroxy, amino, nitro, cyano, C 1-6alkyl, halo-C 1-6 alkyl, hydroxy-C 1-6 alkyl, amino-C 1-6 alkyl, C 1-6 alkoxy, halo-C 1-6 alkoxy, hydroxy-C 1-6 alkoxy or amino-C 1-6 alkoxy.

[0044] In certain embodiments, R 2 , R 3 and the carbon atom to which it is attached together form a 3- to 8-membered cycloalkyl or a 3- to 8-membered heterocyclic group containing 1 to 2 heteroatoms optionally substituted by 1 to 2 Q2, said heteroatoms being selected from N, O or S; each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, C 1-6 alkyl, halo-C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6 alkoxy or halo-C 1-6 alkoxy.

[0045] In certain embodiments, R 2 , R 3 and the carbon atom to which it is attached together form a 3- to 7-membered cycloalkyl or a 3- to 7-membered heterocyclic group optionally substituted by 1 to 2 Q2; each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, C 1-6 alkyl, halo-C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6 alkoxy or halo-C 1-6 alkoxy.

[0046] In certain embodiments, R 2 , R 3 and the carbon atom to which it is attached together form a 3- to 7-membered cycloalkyl or a 3- to 7-membered heterocyclic group containing 1 to 2 heteroatoms optionally substituted by 1 to 2 Q2, said heteroatoms being selected from N, O or S; each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, C 1-6 alkyl, halo-C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6 alkoxy or halo-C 1-6 alkoxy.

[0047] In certain embodiments, R 2 , R 3and, together with the carbon atom to which it is attached, form a 5- to 7-membered cycloalkyl or 5- to 7-membered heterocyclic group optionally substituted with 1 to 2 Q2s; each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, C 1-6 alkyl, halo-C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6 alkoxy or halo-C 1-6 alkoxy.

[0048] In certain embodiments, R 2 , R 3 and, together with the carbon atom to which it is attached, form a 5- to 7-membered cycloalkyl or 5- to 7-membered heterocyclic group containing 1 to 2 heteroatoms optionally substituted with 1 to 2 Q2s, the heteroatoms being selected from N, O or S; each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, C 1-6 alkyl, halo-C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6 alkoxy or halo-C 1-6 alkoxy.

[0049] In certain embodiments, R 2 , R 3 and, together with the carbon atom to which it is attached, form a 5- to 6-membered cycloalkyl or 5- to 6-membered heterocyclic group optionally substituted with 1 to 2 Q2s; each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, C 1-6 alkyl, halo-C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6 alkoxy or halo-C 1-6 alkoxy.

[0050] In certain embodiments, R 2 , R 3 and, together with the carbon atom to which it is attached, form a 5- to 6-membered cycloalkyl or 5- to 6-membered heterocyclic group containing 1 to 2 heteroatoms optionally substituted with 1 to 2 Q2s, the heteroatoms being selected from N, O or S; each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, C 1-6 alkyl, halo-C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6 alkoxy or halo-C 1-6 alkoxy.

[0051] In certain embodiments, R 2 and R 3 together with the carbon atom to which they are attached form a group optionally substituted with 1-2 Q2s as follows:

[0052]

[0053] Each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, C 1-6 alkyl, halo C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6 alkoxy or halo C 1-6 alkoxy.

[0054] In certain embodiments, R 1 is selected from H, halogen, hydroxy, amino, nitro, cyano, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, amino C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 alkoxy or amino C 1-6 alkoxy;

[0055] R 2 and R 3 are each independently selected from H, halogen, hydroxy, amino, nitro, cyano, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, amino C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 alkoxy or amino C 1-6 alkoxy;

[0056] Or R 2 and R 3 together with the carbon atom to which they are attached form a group optionally substituted with 1-2 Q2s as follows:

[0057]

[0058] Each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, C 1-6 alkyl, halo C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6Alkoxy or halo C 1-6 Alkoxy;

[0059] R 4 is H; R 5 and R 6 are each independently selected from H, halogen, hydroxy, amino, nitro, cyano, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, amino C 1-6 alkyl, C 1-6 alkoxy or halo C 1-6 alkoxy;

[0060] R 7 is selected from H, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl or amino C 1-6 alkyl.

[0061] In certain embodiments, R 2 and R 3 together with the carbon atom to which they are attached form one of the following groups optionally substituted with 1-2 Q2 groups:

[0062]

[0063] Each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, methyl, ethyl, propyl, isopropyl, trifluoromethyl, methylamino, ethylamino, dimethylamino, diethylamino, methoxy, ethoxy or trifluoromethoxy.

[0064] In certain embodiments, R 1 is selected from H, halogen, hydroxy, amino, nitro, cyano, methyl, ethyl, propyl, isopropyl, monofluoromethyl, difluoromethyl, trifluoromethyl, hydroxymethyl, aminomethyl, methoxy, ethoxy, propoxy, isopropoxy, monofluoromethoxy, difluoromethoxy or trifluoromethoxy.

[0065] In certain embodiments, R 2 and R 3Independently selected from H, halogen, hydroxyl, amino, nitro, cyano, methyl, ethyl, propyl, isopropyl, trifluoromethyl, trifluoroethyl, trifluoropropyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, aminomethyl, aminoethyl, aminopropyl, methoxy, ethoxy, propoxy, cyclobutyl, cyclopentyl, cyclohexyl, tetrahydrofuran, tetrahydrothienyl, tetrahydropyrrolidinyl, tetrahydropyrazolyl, tetrahydroimidazolyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, piperazinyl, hexahydropyrimidinyl or morpholinyl, each optionally substituted by 1-2 Q1; each Q1 is independently selected from halogen, hydroxyl, amino, nitro, cyano, methyl, ethyl, propyl, isopropyl, trifluoromethyl, methoxy, ethoxy or trifluoromethoxy.

[0066] In certain embodiments, each R 4 and each R 5 and R 6 are independently selected from H, halogen, hydroxyl, amino, nitro, cyano, methyl, ethyl, propyl, isopropyl, trifluoromethyl, methoxy, ethoxy or trifluoromethoxy.

[0067] In certain embodiments, R 5 is selected from H, halogen, hydroxyl, amino, nitro, cyano, methyl, ethyl, propyl, isopropyl, trifluoromethyl, methoxy, ethoxy or trifluoromethoxy.

[0068] In certain embodiments, R 6 is selected from H, halogen, hydroxyl, amino, nitro, cyano, methyl, ethyl, propyl, isopropyl, trifluoromethyl, methoxy, ethoxy or trifluoromethoxy.

[0069] In certain embodiments, R 7 is selected from H, methyl, ethyl, propyl, isopropyl, trifluoromethyl, hydroxymethyl or aminomethyl.

[0070] In certain embodiments, the compound of general formula (I), its pharmaceutically acceptable salt or its isomer further has the structure shown in the following general formula (IIIa) or (IIIb),

[0071]

[0072] wherein, R 1 and R 2 and R 3 and R 7 and Q1 and Q2 are as defined in any one of the technical solutions.

[0073] In certain embodiments, the compound of general formula (I), its pharmaceutically acceptable salt or its isomer further has the following general formula (IIIa),

[0074]

[0075] R 1 is selected from H, halogen, hydroxy, amino, nitro, cyano, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, amino C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 alkoxy or amino C 1-6 alkoxy;

[0076] R 2 , R 3 and the carbon atom to which it is attached together form a 3- to 8-membered cycloalkyl or 3- to 8-membered heterocyclic group optionally substituted by 1 to 2 Q2s; each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, C 1-6 alkyl, halo C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6 alkoxy or halo C 1-6 alkoxy;

[0077] R 7 is selected from H, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl or amino C 1-6 alkyl.

[0078] In certain embodiments, the compound of formula (I), its pharmaceutically acceptable salt or its isomer further has the following formula (IIIa), wherein,

[0079] R 1 is selected from H, halogen, hydroxy, amino, nitro, cyano, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, amino C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 alkoxy or amino C 1-6 alkoxy;

[0080] R 2 , R 3 and the carbon atom to which it is attached together form a 5- to 7-membered cycloalkyl or 5- to 7-membered heterocyclic group optionally substituted by 1 to 2 Q2s; each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, C1-6 alkyl, halo-C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6 alkoxy or halo-C 1-6 alkoxy;

[0081] R 7 is selected from H, C 1-6 alkyl, halo-C 1-6 alkyl, hydroxy-C 1-6 alkyl or amino-C 1-6 alkyl.

[0082] In certain embodiments, the compound of formula (I), its pharmaceutically acceptable salt or its isomer, further has the following general formula (IIIb),

[0083]

[0084] R 1 is selected from H, halogen, hydroxy, amino, nitro, cyano, C 1-6 alkyl, halo-C 1-6 alkyl, hydroxy-C 1-6 alkyl, amino-C 1-6 alkyl, C 1-6 alkoxy, halo-C 1-6 alkoxy, hydroxy-C 1-6 alkoxy or amino-C 1-6 alkoxy;

[0085] R 2 、R 3 and the carbon atom to which it is attached together form a 3- to 8-membered cycloalkyl or 3- to 8-membered heterocyclic group optionally substituted by 1-2 Q2s; each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, C 1-6 alkyl, halo-C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6 alkoxy or halo-C 1-6 alkoxy;

[0086] R 7 is selected from H, C 1-6 alkyl, halo-C 1-6 alkyl, hydroxy-C 1-6 alkyl or amino-C 1-6 alkyl.

[0087] In certain embodiments, the compound of formula (I), its pharmaceutically acceptable salt or its isomer, further has the following general formula (IIIa), wherein,

[0088] R 1 is selected from H, halogen, hydroxy, amino, nitro, cyano, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, amino C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 alkoxy or amino C 1-6 alkoxy;

[0089] R 2 , R 3 and the carbon atom to which it is attached together form a 5- to 7-membered cycloalkyl or 5- to 7-membered heterocyclic group optionally substituted by 1 to 2 Q2; each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, C 1-6 alkyl, halo C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6 alkoxy or halo C 1-6 alkoxy;

[0090] R 7 is selected from H, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl or amino C 1-6 alkyl.

[0091] In certain of the above embodiments, in general formula (IIIa) or (IIIb),

[0092] R 2 , R 3 and the carbon atom to which it is attached together form a 5- to 7-membered cycloalkyl or 5- to 7-membered heterocyclic group containing 1 to 2 heteroatoms optionally substituted by 1 to 2 Q2, the heteroatoms being selected from N, O or S, and the ring atoms of the cycloalkyl or heterocyclic group being optionally oxo;

[0093] each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, C 1-6 alkyl, halo C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6 alkoxy or halo C 1-6 alkoxy.

[0094] In certain of the above embodiments, in general formula (IIIa) or (IIIb),

[0095] R 2 、R 3 and the carbon atom to which it is attached together form the following group optionally substituted by 1 to 2 Q2s:

[0096]

[0097] Each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, C 1-6 alkyl, halo C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6 alkoxy or halo C 1-6 alkoxy.

[0098] In certain of the above embodiments, in general formula (IIIa) or (IIIb),

[0099] R 1 is selected from H, halogen, hydroxy, amino, nitro, cyano, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, amino C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 alkoxy or amino C 1-6 alkoxy;

[0100] R 2 、R 3 and the carbon atom to which it is attached together form the following group optionally substituted by 1 to 2 Q2s:

[0101]

[0102] Each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, C 1-6 alkyl, halo C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6 alkoxy or halo C 1-6 alkoxy;

[0103] R 7 is selected from H, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl or amino C 1-6 alkyl.

[0104] In certain of the above embodiments, in general formula (IIIa) or (IIIb),

[0105] R 1 is selected from H, halogen, hydroxy, amino, nitro, cyano, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, amino C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 alkoxy or amino C 1-6 alkoxy;

[0106] R 2 , R 3 and the carbon atom to which it is attached together form the following group optionally substituted by 1-2 Q2:

[0107]

[0108] Each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, C 1-6 alkyl, halo C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6 alkoxy or halo C 1-6 alkoxy;

[0109] R 7 is selected from H, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl or amino C 1-6 alkyl.

[0110] In certain embodiments, R 1 is selected from H, halogen, hydroxy, amino, nitro, cyano, methyl, ethyl, propyl, isopropyl, monofluoromethyl, difluoromethyl, trifluoromethyl, hydroxymethyl, aminomethyl, methoxy, ethoxy, propoxy, isopropoxy, monofluoromethoxy, difluoromethoxy or trifluoromethoxy;

[0111] R 2 、R 3independently selected from H, halogen, hydroxyl, amino, nitro, cyano, methyl, ethyl, propyl, isopropyl, trifluoromethyl, trifluoroethyl, trifluoropropyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, aminomethyl, aminoethyl, aminopropyl, methoxy, ethoxy, propoxy, cyclobutyl, cyclopentyl, cyclohexyl, tetrahydrofuran, tetrahydrothienyl, tetrahydropyrrolidinyl, tetrahydropyrazolyl, tetrahydroimidazolyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, piperazinyl, hexahydropyrimidinyl or morpholinyl, each optionally substituted by 1-2 Q1; each Q1 is independently selected from halogen, hydroxyl, amino, nitro, cyano, methyl, ethyl, propyl, isopropyl, trifluoromethyl, methoxy, ethoxy or trifluoromethoxy;

[0112] or R 2 , R 3 and the carbon atom to which it is attached together form one of the following groups, each optionally substituted by 1-2 Q2:

[0113]

[0114] each Q2 is independently selected from halogen, hydroxyl, amino, nitro, cyano, carbonyl, methyl, ethyl, propyl, isopropyl, trifluoromethyl, methylamino, ethylamino, dimethylamino, diethylamino, methoxy, ethoxy or trifluoromethoxy;

[0115] R 4 is H; R 5 , R 6 are independently selected from H, halogen, hydroxyl, amino, nitro, cyano, methyl, ethyl, propyl, isopropyl, trifluoromethyl, methoxy, ethoxy or trifluoromethoxy;

[0116] R 7 is selected from H, methyl, ethyl, propyl, isopropyl, trifluoromethyl, hydroxymethyl or aminomethyl.

[0117] In certain embodiments, R 1 is selected from H, halogen, hydroxyl, amino, nitro, cyano, methyl, ethyl, propyl, isopropyl, fluoromethyl, difluoromethyl, trifluoromethyl, hydroxymethyl, aminomethyl, methoxy, ethoxy, propoxy, isopropoxy, fluoromethoxy, difluoromethoxy or trifluoromethoxy;

[0118] R 2 , R 3 and the carbon atom to which it is attached together form one of the following groups, each optionally substituted by 1-2 Q2:

[0119]

[0120] Each Q2 is independently selected from halogen, hydroxyl, amino, nitro, cyano, carbonyl, methyl, ethyl, propyl, isopropyl, trifluoromethyl, methylamino, ethylamino, dimethylamino, diethylamino, methoxy, ethoxy or trifluoromethoxy;

[0121] R 4 is H; R 5 , R 6 are each independently selected from H, halogen, hydroxyl, amino, nitro, cyano, methyl, ethyl, propyl, isopropyl, trifluoromethyl, methoxy, ethoxy or trifluoromethoxy;

[0122] R 7 is selected from H, methyl, ethyl, propyl, isopropyl, trifluoromethyl, hydroxymethyl or aminomethyl.

[0123] In certain embodiments, the compound of general formula (I), its pharmaceutically acceptable salt or its isomer further has the structure shown in the following general formula (II),

[0124]

[0125] wherein, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , X, X1, X2, X3, X4, X5, X6, Q1, Q2 are defined as in any one of the above technical solutions.

[0126] In certain embodiments, the compound of general formula (I), its pharmaceutically acceptable salt or its isomer further has the structure shown in the following general formula (Ia) or (Ib),

[0127]

[0128] wherein, R 1 , R 2 , R 3 , R 5 , R 6 , R 7 , X, X5, X6, Q1, Q2 are defined as in any one of the above technical solutions.

[0129] In certain embodiments, the compound of general formula (I), its pharmaceutically acceptable salt or its isomer further has the structure shown in the following general formula (Ic) or (Id),

[0130]

[0131] Among them, X, R 1 、R 2 、R 3 、R 7 、Q1, Q2 are defined as in any of the above technical solutions.

[0132] In the present invention, the various technical solutions can be combined with each other to form new technical solutions, and the new technical solutions thus formed are also included within the scope of the present invention.

[0133] In certain embodiments, the compound represented by the foregoing general formula (I), its pharmaceutically acceptable salt or its isomer is selected from the following compounds:

[0134]

[0135]

[0136]

[0137] On the other hand, the present invention also provides an intermediate represented by the general formula (V),

[0138]

[0139] Among them, X1, X2, X3, X4, X5, X6, R 1 、R 2 、R 3 、R 4 、R 5 、R 6 、R 7 、Q1, Q2, virtual bond are defined as in any of the above technical solutions; Y is selected from halogen, amino, hydroxyl or mercapto.

[0140] In certain embodiments, the intermediate represented by the general formula (V) further has the structure represented by the following general formula (Va) or (Vb),

[0141]

[0142] Among them, R 2 、R 3 、R 5 、R 6 、R 7 、X5, X6, Q1, Q2 are defined as in any of the above technical solutions; Y is selected from halogen, amino, hydroxyl or mercapto.

[0143] In certain embodiments, the intermediate represented by the general formula (V) further has the structure represented by the following general formula (Vc) or (Vd),

[0144]

[0145] Among them, R 2 、R 3 、R 7 、Q1, and Q2 are as defined in any of the above technical solutions; Y is selected from halogen, amino, hydroxyl, or mercapto.

[0146] In another aspect, the present invention also provides a pharmaceutical preparation, which contains the compound, its pharmaceutically acceptable salt or its isomer described by the foregoing general formula (I), general formula (Ia), general formula (Ib), general formula (Ic), general formula (Id), general formula (II), general formula (IIa), general formula (IIb), general formula (IIIa), or general formula (IIIb), and one or more pharmaceutically acceptable excipients. The pharmaceutical preparation can be any pharmaceutically acceptable dosage form. The pharmaceutically acceptable excipient is a substance that is non-toxic, compatible with the active ingredient, and otherwise biologically suitable for organisms. The selection of a specific excipient will depend on the mode of administration or the type and condition of the disease for treating a specific patient.

[0147] In certain embodiments, the above pharmaceutical preparation can be administered to a patient or subject in need of such treatment by oral, parenteral, rectal, or pulmonary administration, etc. When used for oral administration, the pharmaceutical composition can be made into an oral preparation, for example, it can be made into a conventional oral solid preparation, such as tablets, capsules, pills, granules, etc.; it can also be made into an oral liquid preparation, such as oral solution, oral suspension, syrup, etc. When used for parenteral administration, the above pharmaceutical preparation can also be made into an injection, including injection solution, sterile powder for injection, and concentrated solution for injection. When used for rectal administration, the pharmaceutical composition can be made into suppositories, etc. When used for pulmonary administration, the pharmaceutical composition can be made into inhalation preparations, aerosols, powder aerosols, or sprays, etc.

[0148] In another aspect, the present invention also relates to the use of the compound, its pharmaceutically acceptable salt or its isomer described by the foregoing general formula (I), general formula (Ia), general formula (Ib), general formula (Ic), general formula (Id), general formula (II), general formula (IIa), general formula (IIb), general formula (IIIa), or general formula (IIIb) in the preparation of a drug for preventing and / or treating diseases such as benign tumors or cancers, and the cancers include carcinoma in situ and metastatic cancers.

[0149] Furthermore, the present invention also relates to the use of a pharmaceutical preparation containing the compound, its pharmaceutically acceptable salt or its isomer described by the foregoing general formula (I), general formula (Ia), general formula (Ib), general formula (Ic), general formula (Id), general formula (II), general formula (IIa), general formula (IIb), general formula (IIIa) or general formula (IIIb) in the preparation of a drug for preventing and / or treating diseases such as benign tumors or cancers, wherein the cancers include carcinoma in situ and metastatic cancers.

[0150] On the other hand, the present invention also relates to the use of the compound, its pharmaceutically acceptable salt or its isomer described by the foregoing general formula (I), general formula (Ia), general formula (Ib), general formula (Ic), general formula (Id), general formula (II), general formula (IIa), general formula (IIb), general formula (IIIa) or general formula (IIIb) in the preparation of a drug for preventing and / or treating diseases such as benign tumors or cancers, wherein the drug is used in combination with radiotherapy and / or one or more anti-cancer agents, and the cancers include carcinoma in situ and metastatic cancers.

[0151] Furthermore, the present invention also relates to the use of a pharmaceutical preparation containing the compound, its pharmaceutically acceptable salt or its isomer described by the foregoing general formula (I), general formula (Ia), general formula (Ib), general formula (Ic), general formula (Id), general formula (II), general formula (IIa), general formula (IIb), general formula (IIIa) or general formula (IIIb) in the preparation of a drug for preventing and / or treating diseases such as benign tumors or cancers, wherein the drug can be used in combination with radiotherapy and / or one or more anti-cancer agents, and the cancers include carcinoma in situ and metastatic cancers.

[0152] On the other hand, the present invention also relates to the use of the compound, its pharmaceutically acceptable salt or its isomer described by the foregoing general formula (I), general formula (Ia), general formula (Ib), general formula (Ic), general formula (Id), general formula (II), general formula (IIa), general formula (IIb), general formula (IIIa) or general formula (IIIb) in the preparation of a drug for making cancer cells sensitive to anti-cancer agents and / or radiotherapy.

[0153] Furthermore, the present invention also relates to the use of a pharmaceutical preparation containing the compound, its pharmaceutically acceptable salt or its isomer described by the foregoing general formula (I), general formula (Ia), general formula (Ib), general formula (Ic), general formula (Id), general formula (II), general formula (IIa), general formula (IIb), general formula (IIIa) or general formula (IIIb) in the preparation of a drug for making cancer cells sensitive to anti-cancer agents and / or radiotherapy.

[0154] In another aspect, the present invention also provides a pharmaceutical composition comprising a compound of the foregoing general formula (I), general formula (Ia), general formula (Ib), general formula (Ic), general formula (Id), general formula (II), general formula (IIa), general formula (IIb), general formula (IIIa) or general formula (IIIb), a pharmaceutically acceptable salt thereof or an isomer thereof, and one or more second therapeutic active agents, wherein the second therapeutic active agent is selected from anticancer agents, including mitotic inhibitors, alkylating agents, antimetabolites, DNA intercalators, antitumor antibiotics, growth factor inhibitors, signal transduction inhibitors, cell cycle inhibitors, enzyme inhibitors, retinoid receptor regulators, proteasome inhibitors, topoisomerase inhibitors, biological response modifiers, hormonal drugs, angiogenesis inhibitors, cell growth inhibitors, targeting antibodies, HMG-CoA reductase inhibitors and isoprenyl protein transferase inhibitors.

[0155] In certain embodiments, the second therapeutic active agent may be a drug that alleviates or reduces one or more side effects produced by the compounds of the present invention when used to treat a subject's disease, or may be a drug that enhances the efficacy of the compounds of the present invention.

[0156] In certain embodiments, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients as described above.

[0157] In another aspect, the present invention also relates to the use of a pharmaceutical composition comprising a compound of the foregoing general formula (I), general formula (Ia), general formula (Ib), general formula (Ic), general formula (Id), general formula (II), general formula (IIa), general formula (IIb), general formula (IIIa) or general formula (IIIb), a pharmaceutically acceptable salt thereof or an isomer thereof in the preparation of a drug for preventing and / or treating diseases such as benign tumors or cancers, including carcinoma in situ and metastatic cancers.

[0158] In another aspect, the present invention also relates to the use of a pharmaceutical composition comprising a compound of the foregoing general formula (I), general formula (Ia), general formula (Ib), general formula (Ic), general formula (Id), general formula (II), general formula (IIa), general formula (IIb), general formula (IIIa) or general formula (IIIb), a pharmaceutically acceptable salt thereof or an isomer thereof in the preparation of a drug for preventing and / or treating diseases such as benign tumors or cancers, wherein the drug can be used in combination with radiotherapy and / or one or more anticancer agents, and the cancers include carcinoma in situ and metastatic cancers.

[0159] Furthermore, the present invention also relates to the use of a pharmaceutical composition comprising the compound of the foregoing general formula (I), general formula (Ia), general formula (Ib), general formula (Ic), general formula (Id), general formula (II), general formula (IIa), general formula (IIb), general formula (IIIa) or general formula (IIIb), a pharmaceutically acceptable salt thereof or an isomer thereof, in the preparation of a drug for making cancer cells sensitive to an anticancer agent and / or radiotherapy.

[0160] In another aspect, the present invention also provides a method for treating a disease associated with overactivation of DNA-PK, the method comprising administering to a patient in need an effective amount of the compound of the foregoing general formula (I), general formula (Ia), general formula (Ib), general formula (Ic), general formula (Id), general formula (II), general formula (IIa), general formula (IIb), general formula (IIIa) or general formula (IIIb), a pharmaceutically acceptable salt thereof or an isomer thereof, the foregoing pharmaceutical preparation or pharmaceutical composition; the disease associated with overactivation of DNA-PK is selected from benign tumors or cancers, and the cancers include carcinoma in situ and metastatic cancers.

[0161] Furthermore, the present invention also provides a method for treating a disease associated with overactivation of DNA-PK, the method comprising administering to a patient before / after radiotherapy an effective amount of the compound of the foregoing general formula (I), general formula (Ia), general formula (Ib), general formula (Ic), general formula (Id), general formula (II), general formula (IIa), general formula (IIb), general formula (IIIa) or general formula (IIIb), a pharmaceutically acceptable salt thereof or an isomer thereof, the foregoing pharmaceutical preparation or pharmaceutical composition; the disease associated with overactivation of DNA-PK is selected from benign tumors or cancers, and the cancers include carcinoma in situ and metastatic cancers.

[0162] Furthermore, the present invention also provides a method for treating a disease associated with overactivation of DNA-PK, the method comprising administering to a patient before / after chemotherapy an effective amount of the compound of the foregoing general formula (I), general formula (Ia), general formula (Ib), general formula (Ic), general formula (Id), general formula (II), general formula (IIa), general formula (IIb), general formula (IIIa) or general formula (IIIb), a pharmaceutically acceptable salt thereof or an isomer thereof, the foregoing pharmaceutical preparation or pharmaceutical composition; the disease associated with overactivation of DNA-PK is selected from benign tumors or cancers, and the cancers include carcinoma in situ and metastatic cancers.

[0163] On the other hand, the present invention also provides a method for enhancing the sensitivity of a patient to an anticancer agent or radiotherapy, the method comprising administering to a patient in need thereof an effective amount of the aforementioned compound of general formula (I), general formula (Ia), general formula (Ib), general formula (Ic), general formula (Id), general formula (II), general formula (IIa), general formula (IIb), general formula (IIIa) or general formula (IIIb), a pharmaceutically acceptable salt thereof or an isomer thereof, the aforementioned pharmaceutical preparation or pharmaceutical composition; the anticancer agent is as described below.

[0164] Furthermore, the present invention also provides a method for enhancing the sensitivity of a patient to an anticancer agent or radiotherapy, the method comprising administering to a patient before / after radiotherapy an effective amount of the aforementioned compound of general formula (I), general formula (Ia), general formula (Ib), general formula (Ic), general formula (Id), general formula (II), general formula (IIa), general formula (IIb), general formula (IIIa) or general formula (IIIb), a pharmaceutically acceptable salt thereof or an isomer thereof, the aforementioned pharmaceutical preparation or pharmaceutical composition; the anticancer agent is as described below.

[0165] Furthermore, the present invention also provides a method for enhancing the sensitivity of a patient to an anticancer agent or radiotherapy, the method comprising administering to a patient before / after chemotherapy an effective amount of the aforementioned compound of general formula (I), general formula (Ia), general formula (Ib), general formula (Ic), general formula (Id), general formula (II), general formula (IIa), general formula (IIb), general formula (IIIa) or general formula (IIIb), a pharmaceutically acceptable salt thereof or an isomer thereof, the aforementioned pharmaceutical preparation or pharmaceutical composition; the anticancer agent is as described below.

[0166] On the other hand, the present invention also provides a kit, comprising:

[0167] (a) an effective amount of one or more of the aforementioned compounds of general formula (I), general formula (Ia), general formula (Ib), general formula (Ic), general formula (Id), general formula (II), general formula (IIa), general formula (IIb), general formula (IIIa) or general formula (IIIb), a pharmaceutically acceptable salt thereof or an isomer thereof,

[0168] and (b) an effective amount of one or more anticancer agents.

[0169] The "anticancer agent" referred to in the present invention means an agent having a certain therapeutic effect on tumors, including but not limited to mitotic inhibitors, alkylating agents, antimetabolites, DNA intercalators, antitumor antibiotics, growth factor inhibitors, signal transduction inhibitors, cell cycle inhibitors, enzyme inhibitors, retinoid receptor regulators, proteasome inhibitors, topoisomerase inhibitors, biological response modifiers, hormonal drugs, angiogenesis inhibitors, cell growth inhibitors, targeting antibodies, HMG-CoA reductase inhibitors, isoprenyl protein transferase inhibitors, etc.; the tumors include benign tumors and cancers. The "effective amount" refers to the dose of a drug that can prevent, alleviate, delay, inhibit or cure the disease of a subject. The size of the administered dose is related to the drug administration method, the pharmacokinetics of the agent, the severity of the disease, the individual signs of the subject (gender, weight, height, age), etc.

[0170] In the present invention, unless otherwise specified, scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. However, for a better understanding of the present invention, the definitions of some terms are provided below. When the definitions and explanations of the terms provided by the present invention are inconsistent with the meanings commonly understood by those skilled in the art, the definitions and explanations provided by the present invention shall prevail.

[0171] The "halogen" referred to in the present invention means a fluorine atom, a chlorine atom, a bromine atom or an iodine atom.

[0172] The "C 1-6 alkyl" in the present invention means a straight-chain or branched-chain alkyl containing 1-6 carbon atoms, including, for example, "C 1-4 alkyl", "C 1-3 alkyl", "C 1-2 alkyl", "C 2-6 alkyl", "C 2-5 alkyl", "C 2-4 alkyl", "C 2-3 alkyl", "C 3-6 alkyl", "C 3-5 alkyl", "C 3-4 alkyl", etc. Specific examples include but are not limited to: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, 2-methylbutyl, neopentyl, 1-ethylpropyl, n-hexyl, isohexyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-ethylbutyl, 1,2-dimethylpropyl, etc. The "C 1-4 alkyl" in the present invention refers to C 1-6Specific examples of alkyl groups containing 1 to 4 carbon atoms.

[0173] As used in the present invention, "C 1-6 alkoxy" means "C 1-6 alkyl - O -", and the "C 1-6 alkyl" is as defined above. As used in the present invention, "C 1-4 alkoxy" means "C 1-4 alkyl - O -", and the "C 1-4 alkyl" is as defined above.

[0174] As used in the present invention, "C 1-6 alkylthio" means "C 1-6 alkyl - S -", and the "C 1-6 alkyl" is as defined above. As used in the present invention, "C 1-4 alkylthio" means "C 1-4 alkyl - S -", and the "C 1-4 alkyl" is as defined above.

[0175] As used in the present invention, "hydroxy C 1-6 alkyl, amino C 1-6 alkyl, halo C 1-6 alkyl" means that one or more hydrogens in C 1-6 alkyl are each replaced by one or more hydroxyl groups, amino groups, or halogens. C 1-6 alkyl is as defined above

[0176] As used in the present invention, "hydroxy C 1-6 alkoxy, amino C 1-6 alkoxy, halo C 1-6 alkoxy" means that one or more hydrogens in "C 1-6 alkoxy" are replaced by one or more hydroxyl groups, amino groups, or halogens.

[0177] As used in the present invention, "hydroxy C 1-6 alkylthio, amino C 1-6 alkylthio, halo C 1-6 alkylthio" means that one or more hydrogens in "C 1-6 alkylthio" are replaced by one or more hydroxyl groups, amino groups, or halogens.

[0178] As used in the present invention, "C 1-6 alkylamino, di(C 1-6 alkyl)amino" mean C 1-6 alkyl - NH - and

[0179] The "3-8 membered cycloalkyl" as used in the present invention refers to a saturated or partially saturated monocyclic group containing 3-8 ring atoms and having no aromaticity. The "3-8 membered cycloalkyl" as used in the present invention includes "3-8 membered saturated cycloalkyl" and "3-8 membered partially saturated cycloalkyl", for example, it is "3-6 membered cycloalkyl", "3-6 membered saturated cycloalkyl", "5-7 membered cycloalkyl", "5-7 membered saturated cycloalkyl", "5-6 membered cycloalkyl", "5-6 membered saturated cycloalkyl", etc. Its examples include but are not limited to: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclohexenyl, etc.

[0180] The "3-8 membered heterocyclic group" as used in the present invention refers to a saturated or partially saturated monocyclic group containing at least one (for example, containing 1, 2, 3, 4 or 5) heteroatom and having 3-8 ring atoms, and the heteroatom is a nitrogen atom, an oxygen atom and / or a sulfur atom. Optionally, the ring atoms (such as carbon atoms, nitrogen atoms or sulfur atoms) in the cyclic structure can be oxo-substituted. The "3-8 membered heterocyclic group" as used in the present invention includes "3-8 membered saturated heterocyclic group" and "3-8 membered partially saturated heterocyclic group". The "3-8 membered heterocyclic group" is, for example, "3-6 membered heterocyclic group", "3-6 membered saturated heterocyclic group", "3-7 membered heterocyclic group", "3-7 membered saturated heterocyclic group", "5-7 membered heterocyclic group", "5-7 membered saturated heterocyclic group", "5-6 membered heterocyclic group", "5-6 membered saturated heterocyclic group", etc. Its specific examples include but are not limited to: aziridinyl, 2H-aziridinyl, diaziridinyl, 3H-diazirinyl, azetidinyl, oxiranyl, oxetanyl, 1,4-dioxanyl, 1,3-dioxanyl, 1,3-dioxolanyl, 1,4-dioxadienyl, tetrahydrofuryl, dihydropyrrolyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, 4,5-dihydroimidazolyl, pyrazolidinyl, 4,5-dihydropyrazolyl, 2,5-dihydrothienyl, tetrahydrothienyl, 4,5-dihydrothiazolyl, thiazolidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, 1,1-dioxotetrahydrothiopyranyl, piperidinyl, tetrahydropyridyl, piperidone, tetrahydropyridone, dihydropiperidone, piperazinyl, hexahydropyrimidinyl, morpholinyl, etc.

[0181] The "C(R 4 )", "CH(R 5 )", "C(R 6 )", "N(R 7 )" as used in the present invention respectively refer to the following structures:

[0182]

[0183] As used in the present invention When it appears in a group, such as then it indicates the connection position of the group to the adjacent group; when it appears at the position of a chemical bond in a chemical structure, especially at the position of a substituent on a 6-membered saturated ring, such as then it represents a chemical bond and is specifically located on the a-bond (axial bond) or e-bond (equatorial bond) of a chair-form 6-membered saturated ring.

[0184] "Each R 4 " as described in the present invention means that when at least two of X1-X4 are selected from "C(R 4 )", each R 4 in the multiple Rs 4 is independently selected from the groups described in the above technical solutions.

[0185] "Each R 5 " as described in the present invention means that when X5 and X6 are both CH(R 5 ) at the same time, each R 5 in the multiple Rs 5 is independently selected from the groups described in the above technical solutions.

[0186] "Each R 7 " as described in the present invention means that when X5 and X6 are both N(R 7 ) at the same time, each R 7 in the multiple Rs 7 is independently selected from the groups described in the above technical solutions.

[0187] "Optionally substituted by a substituent" as described in the present invention means two situations where one or more hydrogen atoms on the group to be substituted are "substituted" or "not substituted" by one or more substituents.

[0188] "Chemotherapy" as described in the present invention is an abbreviation for chemical drug therapy, and mainly achieves the purpose of treatment by using chemotherapeutic drugs to kill cancer cells.

[0189] "Radiotherapy" as described in the present invention refers to a tumor treatment method, namely tumor radiotherapy, which mainly uses radiation for local treatment of tumors. The "radiation" mentioned includes α, β, γ rays generated by radioactive isotopes and x-rays, electron beams, proton beams, and other particle beams generated by various x-ray therapy machines or accelerators.

[0190] The "pharmaceutically acceptable salts" referred to in the present invention mean salts formed by acidic functional groups (such as -COOH, -OH, -SO3H, etc.) present in the compound with appropriate inorganic or organic cations (bases), including salts formed with alkali metals or alkaline earth metals, ammonium salts, salts formed with nitrogen-containing organic bases; and salts formed by basic functional groups (such as -NH2, etc.) present in the compound with appropriate inorganic or organic anions (acids), including salts formed with inorganic acids or organic acids (such as carboxylic acids, etc.).

[0191] The "isomers" referred to in the present invention mean that the compounds of the present invention contain one or more asymmetric centers, and thus can exist as racemates and racemic mixtures, single enantiomers, diastereomer mixtures and single diastereomers. The compounds of the present invention may have asymmetric centers, and such asymmetric centers independently generate two optical isomers each. The scope of the present invention includes all possible optical isomers and their mixtures. If the compounds of the present invention contain olefin double bonds, unless otherwise specified, they include cis isomers and trans isomers. The compounds of the present invention may exist in the form of tautomers (a kind of functional group isomers), which have different hydrogen attachment points through the displacement of one or more double bonds. For example, a ketone and its enol form are keto-enol tautomers. The compounds of the present invention contain a spiro ring structure. Affected by the steric structure of the ring, the substituents on the ring may exist on both sides of the ring to form relative cis and trans isomers. Each tautomer and its mixture are included in the scope of the present invention. All enantiomers, diastereomers, racemates, meso forms, cis-trans isomers, tautomers, geometric isomers, epimers and their mixtures of the compounds are included in the scope of the present invention.

[0192] The compounds of the present invention can be prepared in the form of individual enantiomers by enantioselective synthesis or resolution from a mixture of enantiomers. Conventional resolution techniques include using various well-known chromatographic methods to resolve mixtures of enantiomers of the starting material or the final product.

[0193] When the stereochemistry of the disclosed compound is named or depicted by structure, the named or depicted stereoisomer is at least 60% by weight, 70% by weight, 80% by weight, 90% by weight, 99% by weight or 99.9% by weight pure relative to other stereoisomers. When a single isomer is named or depicted by structure, the depicted or named enantiomer is at least 60% by weight, 70% by weight, 80% by weight, 90% by weight, 99% by weight or 99.9% by weight pure. The weight % of optical purity is the ratio of the weight of the enantiomer to the weight of the enantiomer plus the weight of its optical isomer.

[0194] Advantages of the Invention

[0195] 1. The compounds of the present invention, their pharmaceutically acceptable salts or their isomers have excellent DNA-PK inhibitory effects, have good pharmacokinetic properties in vivo (such as in mice, rats, dogs, monkeys, humans, etc.), have a long-lasting effect, high bioavailability, and can enhance the sensitivity of cancer cells (such as lung cancer cell line A549, liver cancer cell line huh-7, breast cancer cell lines MDA-MB-231 and MCF-7, ovarian cancer cell line SKOV3, etc.) to radiotherapy and / or one or more anticancer agents (such as chemotherapy drugs).

[0196] 2. The compounds of the present invention, their pharmaceutically acceptable salts or their isomers have good therapeutic effects on benign tumors and cancers, and have high stability in liver microsomes in multiple species (such as human, murine, monkey, and dog).

[0197] 3. The preparation process of the compounds of the present invention is simple, the drug purity is high, the quality is stable, and it is easy to carry out large-scale industrial production.

[0198] 4. The anti-tumor effect of the compounds of the present invention in vivo (such as in CDX models in nude mice: NCI-H1048 model and MDA-MB-231 model) is significant, and can significantly improve the tumor inhibition rate of radiotherapy and / or one or more anticancer agents (such as chemotherapy drugs). Specific Embodiments

[0199] The technical solutions of the present invention will be described below in conjunction with specific embodiments. The above content of the present invention will be further described in detail, but it should not be understood that the scope of the above subject matter of the present invention is limited to the following embodiments. All technologies implemented based on the above content of the present invention belong to the scope of the present invention.

[0200] Abbreviations:

[0201] BrettPhos Pd G3: Palladium(II) (2-dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl) methanesulfonate; DMA: N,N-dimethylacetamide; DCM: Dichloromethane; MeOH: Methanol; PE: Petroleum ether; EA: Ethyl acetate; THF: Tetrahydrofuran; DIEA: N,N-diisopropylethylamine; S-CDI: N,N'-thiocarbonyldiimidazole; DMAP: 4-Dimethylaminopyridine.

[0202] Preparation Example 1: Preparation of 2-chloro-2',3',5',6,6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-pyran]

[0203] 1. Preparation of 4-((2-chloro-5-nitropyrimidin-4-yl)amino)tetrahydro-2H-pyran-4-carbonitrile

[0204]

[0205] 2,4-Dichloro-5-nitropyrimidine (7.76 g, 40.0 mmol) and DIEA (10.4 g, 80.5 mmol) were added to THF (150 mL). 4-Aminotetrahydro-2H-pyran-4-carbonitrile (5.05 g, 40.0 mmol) was added at -20 °C, and then the reaction was carried out at 25 °C for 2 hours. Then the system was cooled to 20 °C, concentrated and purified by silica gel column chromatography (PE:EA = 2:1) to obtain the product (10.2 g, yield 89.9%).

[0206] Preparation of 4-((5-amino-2-chloropyrimidin-4-yl)amino)tetrahydro-2H-pyran-4-carbonitrile

[0207]

[0208] 4-((2-Chloro-5-nitropyrimidin-4-yl)amino)tetrahydro-2H-pyran-4-carbonitrile (9.8 g, 34.5 mmol) and iron powder (7.7 g, 137.5 mmol) were added to a mixed system of HOAc (20 mL), ethanol (20 mL) and water (20 mL), and then the reaction was carried out at 70 °C for 1 hour. Then the system was cooled to 25 °C, and then filtered under reduced pressure. The filter cake was washed with (DCM:MeOH = 10:1) (20 mL). The obtained filtrate was concentrated by rotary evaporation and purified by silica gel column chromatography (DCM:MeOH = 15:1) to obtain the product (8.0 g, yield 91.4%).

[0209] Preparation of 4-(2-chloro-8-thioxo-7,8-dihydro-9H-purin-9-yl)tetrahydro-2H-pyran-4-carbonitrile

[0210]

[0211] 4-((5-Amino-2-chloropyrimidin-4-yl)amino)tetrahydro-2H-pyran-4-carbonitrile (4.7 g, 18.5 mmol), imidazole (2.5 g, 36.7 mmol) and S-CDI (6.5 g, 36.5 mmol) were added to DCM (90 mL), and then the system was reacted at 25 °C for 1 hour. Then the pH of the system was adjusted to 5 with 2N HCl (40 mL). A solid precipitated from the system, and then it was filtered under reduced pressure. The filter cake was dried to obtain the product (4.5 g, yield 82.3%).

[0212] Preparation of 2-chloro-2',3',5',6,6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-pyran]

[0213]

[0214] 4-(2-chloro-8-thioxo-7,8-dihydro-9H-purin-9-yl)tetrahydro-2H-pyran-4-carbonitrile (4.0 g, 13.5 mmol) was added to THF (80 mL), and then lithium aluminum hydride (1.5 g, 39.5 mmol) was added portionwise at 40 °C. The reaction was then carried out at 40 °C for 0.5 h. The reaction system was cooled to 20 °C, and water (2 mL) was slowly added to quench the reaction. Then (DCM:MeOH = 10:1) (50 mL) was added and stirred for 0.5 h. After that, filtration was carried out under reduced pressure, and the filter cake was washed with (DCM:MeOH = 10:1) (50 mL). The obtained filtrate was concentrated and purified by silica gel column chromatography (DCM:MeOH = 20:1) to obtain the product (1.3 g, yield 36.2%).

[0215] Example 1: Preparation of 5-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-2',3',5,5',6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-pyran]-2-amine (Compound 1)

[0216] 1. Preparation of 2-chloro-5-methyl-2',3',5,5',6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-pyran] (Compound 1a)

[0217]

[0218] 2-Chloro-2',3',5',6,6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-pyran] (266 mg, 1 mmol) and methyl iodide (2.85 g, 20.0 mmol) were added to acetonitrile (10 mL), and the reaction system was then reacted at 105 °C under microwave for 1 h. Finally, the reaction system was concentrated and purified by silica gel column chromatography (DCM:MeOH = 20:1) to obtain the product (130 mg, yield 46.5%).

[0219] 1 HNMR (400 MHz, d-DMSO): δ 7.91 (s, 1H), 4.10 (s, 2H), 3.98 - 3.92 (m, 2H), 3.44 - 3.40 (m, 2H), 3.27 (s, 3H), 2.31 - 2.23 (m, 2H), 1.81 - 1.74 (m, 2H).

[0220] 1D NOESY (400 MHz, d-DMSO): Excitation peak (upward): 7.914 ppm; Response peak (downward): 3.271 ppm.

[0221] 2. Preparation of 5-Methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-2',3',5,5',6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-pyran]-2-amine

[0222]

[0223] Dissolve 2-chloro-5-methyl-2',3',5,5',6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-pyran] (110 mg, 0.39 mmol), 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (64 mg, 0.43 mmol), cesium carbonate (254 mg, 0.78 mmol) and BrettPhos Pd G3 (35 mg, 0.039 mmol) in 1,4-dioxane (15 mL). Then the reaction system is heated to 105 °C under N2 for 2 hours. After that, the system is cooled to 20 °C and concentrated. The product is purified by silica gel column chromatography (DCM:MeOH = 10:1) to obtain the product (112 mg, yield 73.4%).

[0224] Molecular formula: C 19 H 21 N9O Molecular weight: 391.4 LC-MS (M / e): 392.2 (M+H + )

[0225] 1 1H NMR (400 MHz, d-DMSO): δ 9.13 (s, 1H), 8.45 (s, 1H), 8.36 (s, 1H), 7.72 - 7.68 (m, 2H), 4.08 (s, 2H), 3.97 - 3.92 (m, 2H), 3.44 - 3.33 (m, 2H), 3.23 (s, 3H), 2.36 (s, 3H), 2.35 - 2.31 (m, 2H), 1.80 - 1.72 (m, 2H).

[0226] Example 2: Preparation of 6-Methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-2',3',5',6,6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-pyran]-2-amine (Compound 2)

[0227] 1. Preparation of 2-chloro-6-methyl-2',3',5',6,6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-pyran] (Compound 2a)

[0228]

[0229] 2-Chloro-2',3',5',6,6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-pyran] (100 mg, 0.38 mmol) was added to THF (12 mL). Then, NaH (60%, 31 mg, 0.78 mmol) was added to the system at 0 °C. After reacting for 20 minutes, methyl iodide (271 mg, 1.9 mmol) was added, and then the reaction was carried out at 20 °C for 1 hour. Finally, the system was concentrated and purified by silica gel column chromatography (DCM:MeOH = 20:1) to obtain the product (100 mg, yield 94.1%).

[0230] 1 HNMR (400 MHz, d-DMSO): 8.27 (s, 1H), 4.05 - 3.97 (m, 4H), 3.52 - 3.42 (m, 2H), 3.01 (s, 3H), 2.35 - 2.25 (m, 2H), 1.96 - 1.88 (m, 2H).

[0231] 1D NOESY (400 MHz, d-DMSO): Excitation peak (upward): 3.014 ppm; Response peak (downward): 4.012 ppm.

[0232] 2. Preparation of 6-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-2',3',5',6,6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-pyran]-2-amine

[0233]

[0234] 2-Chloro-6-methyl-2',3',5',6,6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-pyran] (90 mg, 0.32 mmol), 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (48 mg, 0.32 mmol), cesium carbonate (208.5 mg, 0.64 mmol) and BrettPhos Pd G3 (29 mg, 0.032 mmol) were dissolved in 1,4-dioxane (15 mL). Then, the system was reacted at 105 °C for 2 hours under a N2 atmosphere. After the system was cooled to 20 °C and concentrated, it was purified by silica gel column chromatography (DCM:MeOH = 15:1) to obtain the product (40 mg, yield 32.0%).

[0235] Molecular formula: C 19 H 21 N9O Molecular weight: 391.4 LC-MS (M / e): 392.2 (M + H + )

[0236] 1 HNMR (400 MHz, CDCl3): δ 9.79 (s, 1H), 8.34 (s, 1H), 8.25 (s, 1H), 7.57 (s, 1H), 6.64 (s, 1H), 4.28 - 4.22 (m, 2H), 3.88 (s, 2H), 3.63 - 3.53 (m, 2H), 3.11 (s, 3H), 2.69 - 2.61 (m, 2H), 2.52 (s, 3H), 1.93 - 1.87 (m, 2H).

[0237] Example 3: Preparation of N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-2',3',5',6,6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-pyran]-2-amine (Compound 3)

[0238] 1. Preparation of tert-butyl 2-chloro-2',3',5',6'-tetrahydrospiro[imidazo[1,2-e]purine-8,4'-pyran]-6(7H)-carboxylate

[0239]

[0240] 2-Chloro-2',3',5',6,6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-pyran] (200 mg, 0.75 mmol), Boc2O (328 mg, 1.5 mmol), and DMAP (184 mg, 1.5 mmol) were added to THF (15 mL), and then the reaction was carried out at 25 °C for 2 hours. Finally, the system was concentrated and purified by silica gel column chromatography (DCM:MeOH = 35:1) to obtain the product (250 mg, yield 91.1%).

[0241] 2. Preparation of tert-butyl 2-((7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)amino)-2',3',5',6'-tetrahydrospiro[imidazo[1,2-e]purine-8,4'-pyran]-6(7H)-carboxylate

[0242]

[0243] tert-Butyl 2-chloro-2',3',5',6'-tetrahydrospiro[imidazo[1,2-e]purine-8,4'-pyran]-6(7H)-carboxylate (220 mg, 0.60 mmol), 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (98 mg, 0.66 mmol), cesium carbonate (391 mg, 1.2 mmol) and BrettPhos Pd G3 (55 mg, 0.061 mmol) were dissolved in 1,4-dioxane (15 mL). Then the reaction system was heated to 105 °C under N2 for 3 h. After cooling to 20 °C, the reaction mixture was concentrated and purified by silica gel column chromatography (DCM:MeOH = 30:1) to give the product (260 mg, yield 90.8%).

[0244] 3. Preparation of N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-2',3',5',6,6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-pyran]-2-amine

[0245]

[0246] tert-Butyl 2-((7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)amino)-2',3',5',6'-tetrahydrospiro[imidazo[1,2-e]purine-8,4'-pyran]-6(7H)-carboxylate (230 mg, 0.48 mmol) was added to DCM (6 mL), followed by TFA (3 mL). The reaction system was stirred at 25 °C for 4 h. Then the reaction mixture was concentrated, and the crude product was adjusted to alkaline with saturated sodium bicarbonate solution (1 mL), concentrated again and purified by silica gel column chromatography (DCM:MeOH = 15:1) to give the product (124 mg, yield 68.5%).

[0247] Molecular formula: C 18 H 19 N9O Molecular weight: 377.4 LC-MS (M / e): 378.2 (M+H + )

[0248] 1 1H NMR (400 MHz, d-DMSO): δ 9.19 (s, 1H), 8.75 (s, 1H), 8.39 (s, 1H), 8.10 (s, 1H), 8.03 (s, 1H), 7.72 (s, 1H), 4.04 - 3.94 (m, 4H), 3.58 - 3.53 (m, 2H), 3.43 - 3.33 (m, 5H), 1.91 - 1.87 (m, 2H).

[0249] Example 4: Preparation of 5'-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-5',7'-dihydrospiro[cyclohexane-

[0250] 1,8'-imidazo[1,2-e]purin]-2'-amine (Compound 4) 1. Preparation of 1-aminocyclohexane-1-carbonitrile

[0251]

[0252] Dissolve cyclohexanone (2.7 g, 27.5 mmol) in a solution of ammonia in THF (40 ml), add titanium tetraisopropoxide (6.0 mL), stir at 25 °C for 6 hours, cool to -10 °C, add TMSCN (2.7 g, 27.5 mmol), warm to 0 °C and stir for 16 hours. Filter, extract the filtrate with DCM (100 mL), and rotary evaporate the organic phase to obtain the product (3.1 g), which is directly used in the next step of the reaction.

[0253] 2. Preparation of 1-((2-chloro-5-nitropyrimidin-4-yl)amino)cyclohexane-1-carbonitrile

[0254]

[0255] Dissolve 2,4-dichloro-5-nitropyrimidine (4.5 g, 23.3 mmol) in THF (100 mL), stir at 0 °C, add 1-aminocyclohexane-1-carbonitrile (2.9 g, 23.3 mmol), DIEA (6.0 g, 46.6 mmol), warm to 20 °C and continue stirring for 1 hour. Rotary evaporate and purify by silica gel column chromatography (PE / EA = 8 / 1) to obtain the product (1.6 g, yield 24.3%).

[0256] 3. Preparation of 1-((5-amino-2-chloropyrimidin-4-yl)amino)cyclohexane-1-carbonitrile

[0257]

[0258] Dissolve 1-((2-chloro-5-nitropyrimidin-4-yl)amino)cyclohexane-1-carbonitrile (1.6 g, 5.7 mmol) in EtOH (10 ml), add iron powder (1.3 g, 22.8 mmol), glacial acetic acid (10 mL), and water (10 mL), and warm to 70 °C and stir for 1 hour. Filter, add water (50 mL), adjust the pH > 7 with aqueous NaOH solution, and extract with DCM (100 mL). Rotary evaporate the organic phase and purify by silica gel column chromatography (DCM / MeOH = 10 / 1) to obtain the product (800 mg, yield 56.0%).

[0259] 4. Preparation of 1-(2-chloro-8-thioxo-7,8-dihydro-9H-purin-9-yl)cyclohexane-1-carbonitrile

[0260]

[0261] Dissolve 1-((5-amino-2-chloropyrimidin-4-yl)amino)cyclohexane-1-carbonitrile (800 mg, 3.2 mmol) in DCM (30 mL), add imidazole (361 mg, 5.3 mmol) and S-CDI (944 mg, 5.3 mmol), stir at 25 °C for 1 hour, filter to obtain the product (650 mg, yield 69.6%).

[0262] 5. Preparation of 2'-chloro-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purine]

[0263]

[0264] Dissolve 1-(2-chloro-8-thioxo-7,8-dihydro-9H-purin-9-yl)cyclohexane-1-carbonitrile (620 mg, 2.1 mmol) in THF (20 ml), add LiAlH4 (340 mg, 6.3 mmol), stir at 40 °C for 15 minutes, add water to quench the reaction, spin-dry and purify by silica gel column chromatography (DCM / MeOH = 20 / 1) to obtain the product (200 mg, yield 35.9%).

[0265] 6. Preparation of 2'-chloro-5'-methyl-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purine]

[0266]

[0267] Dissolve 2'-chloro-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purine] (150 mg, 0.57 mmol) in acetonitrile (10 mL), add methyl iodide (1.6 g, 11.4 mmol), stir at 105 °C by microwave for 1.5 hours, spin-dry and purify by silica gel column chromatography (DCM / MeOH = 94 / 6) to obtain the product (90 mg, yield 57.0%).

[0268] 7. Preparation of 5'-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purin]-2'-amine

[0269]

[0270] (2'-Chloro-5'-methyl-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purine] (90 mg, 0.32 mmol) was dissolved in dioxane (30 mL), and 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (52 mg, 0.35 mmol), Brettphos Pd G3 (29 mg, 0.032 mmol), and Cs2CO3 (209 mg, 0.64 mmol) were added. The mixture was stirred at 110 °C for 24 hours under nitrogen protection. The solvent was evaporated, and the residue was purified by silica gel column chromatography (DCM / MeOH = 20 / 1) to obtain the product (45 mg, yield 35.7%).

[0271] Molecular formula: C 20 H 23 N9 Molecular weight: 389.21 LC-MS (M / e): 390.2 (M + H + )

[0272] 1 1H-NMR (400 MHz, CDCl3) δ: 9.14 (s, 1H), 8.53 (s, 1H), 8.38 (s, 1H), 7.80 (s, 1H), 7.70 (s, 1H), 3.98 (s, 2H), 3.27 (s, 3H), 2.39 (s, 3H), 2.19 - 2.09 (m, 2H), 1.82 - 1.76 (m, 4H), 1.67 - 1.58 (m, 1H), 1.34 - 1.14 (m, 3H).

[0273] Example 5: Preparation of 4,4-difluoro-5'-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purin]-2'-amine (Compound 5)

[0274] 1. Preparation of 2'-chloro-4,4-difluoro-5'-methyl-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purine]

[0275]

[0276] 2'-Chloro-5'-methyl-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purine]-4-one (100.0 mg, 0.34 mmol) was dissolved in DCM (5.0 mL), and DAST (109.6 mg, 0.68 mmol) was added. The mixture was stirred at 16 °C for 2 hours. The solvent was evaporated, and the residue was purified by silica gel column chromatography (DCM / MeOH = 20 / 1) to obtain the product (60.0 mg, yield 55.8%).

[0277] 2. Preparation of 4,4 - Difluoro - 5'-Methyl - N-(7 - Methyl - [1,2,4]Triazolo[1,5 - a]Pyridin - 6 - yl)-5',7'-Dihydrospiro[Cyclohexane - 1,8'-Imidazo[1,2 - e]Purine]-2'-Amine

[0278]

[0279] Dissolve 2'-Chloro - 4,4 - difluoro - 5'-methyl - 5',7'-dihydrospiro[cyclohexane - 1,8'-imidazo[1,2 - e]purine] (50.0 mg, 0.16 mmol) in dioxane (3 mL), add 7 - methyl - [1,2,4]triazolo[1,5 - a]pyridin - 6 - amine (23.7 mg, 0.16 mmol), Brettphos Pd G3 (18.2 mg, 0.02 mmol), Cs2CO3 (104.3 mg, 0.32 mmol), and stir at 100 °C for 2 hours under nitrogen protection. Rotate to dry the solvent and purify by high - pressure preparation (acetonitrile / water) to obtain the product (4.5 mg, yield 6.6%).

[0280] Molecular formula: C 20 H 21 F2N9 Molecular weight: 425.4 LC - MS (M / e): 426.2 (M + H + )

[0281] 1 1H - NMR (400 MHz, CDCl3) δ: 9.68 (s, 1H), 8.27 (s, 1H), 7.60 (s, 1H), 7.57 (s, 1H), 6.55 (s, 1H), 4.15 (s, 2H), 3.36 (s, 3H), 2.53 - 2.34 (m, 7H), 2.06 - 1.93 (m, 4H).

[0282] Example 6: Preparation of 5'-Methyl - 2'-((7 - Methyl - [1,2,4]Triazolo[1,5 - a]Pyridin - 6 - yl)Amino)-5',7'-Dihydrospiro[Cyclohexane - 1,8'-Imidazo[1,2 - e]Purine]-4 - One (Compound 6)

[0283] 1. Preparation of 8 - Amino - 1,4 - Dioxaspiro[4.5]Decane - 8 - Carbonitrile

[0284]

[0285] Dissolve 1,4-dioxaspiro[4.5]decan-8-one (23.0 g, 147.2 mmol) in methanol (40 ml), add 7M ammonia / methanol (115.7 mL, 809.6 mmol), stir at 25 °C for 4 hours, add TMSCN (14.6 g, 147.2 mmol), stir at -10 °C for 12 hours, and rotary evaporate to obtain the product (26.0 g, yield 97.0%).

[0286] 2. Preparation of 8-((2-chloro-5-nitropyrimidin-4-yl)amino)-1,4-dioxaspiro[4.5]decan-8-carbonitrile

[0287]

[0288] Dissolve 2,4-dichloro-5-nitropyrimidine (25.2 g, 129.8 mmol) in THF (400 mL), stir at 0 °C, add 8-amino-1,4-dioxaspiro[4.5]decan-8-carbonitrile (26.0 g, 142.8 mmol), DIEA (33.6 g, 259.6 mmol), stir at 20 °C for 2 hours, rotary evaporate and purify by silica gel column chromatography (PE / EA = 8 / 1 - 3 / 1) to obtain the product (40.0 g, yield 90.7%).

[0289] 3. Preparation of 8-((5-amino-2-chloropyrimidin-4-yl)amino)-1,4-dioxaspiro[4.5]decan-8-carbonitrile

[0290]

[0291] Dissolve 8-((2-chloro-5-nitropyrimidin-4-yl)amino)-1,4-dioxaspiro[4.5]decan-8-carbonitrile (38.0 g, 111.8 mmol) in EtOH (300 ml), add iron powder (25.0 g, 447.2 mmol), glacial acetic acid (300 mL), water (300 mL), heat to 70 °C and stir for 1 hour. Filter, add aqueous sodium carbonate solution to adjust pH > 7, and extract with EA (600 mL). Rotary evaporate the organic phase and purify by silica gel column chromatography (DCM / MeOH = 80 / 1 - 40 / 1) to obtain the product (26 g, yield 74.9%).

[0292] 4. Preparation of 8-(2-chloro-8-thioxo-7,8-dihydro-9H-purin-9-yl)-1,4-dioxaspiro[4.5]decan-8-carbonitrile

[0293]

[0294] 8-((5-Amino-2-chloropyrimidin-4-yl)amino)-1,4-dioxaspiro[4.5]decane-8-carbonitrile (25 g, 80.6 mmol) was dissolved in DCM (400 mL), imidazole (11 g, 161.2 mmol) and S-CDI (28.8 g, 161.2 mmol) were added, and the mixture was stirred at 25 °C for 6 hours. The product was obtained by filtration (16 g, yield 56.3%).

[0295] 5. Preparation of 2-chloro-5,7-dihydrodispiro[imidazo[1,2-e]purine-8,1'-cyclohexane-4',2''-[1,3]dioxolane]

[0296]

[0297] 8-(2-Chloro-8-thioxo-7,8-dihydro-9H-purin-9-yl)-1,4-dioxaspiro[4.5]decane-8-carbonitrile (10 g, 28.4 mmol) was dissolved in THF (100 ml), LiAlH4 (3.2 g, 85.2 mmol) was added, and the mixture was stirred at 40 °C for 1 hour. The reaction was quenched by adding water, and the solvent was evaporated. The product was purified by silica gel column chromatography (DCM / MeOH = 60 / 1 - 20 / 1) to obtain the product (5.6 g, yield 61.3%).

[0298] 6. Preparation of 2-chloro-5-methyl-5,7-dihydrodispiro[imidazo[1,2-e]purine-8,1'-cyclohexane-4',2''-[1,3]dioxolane]

[0299]

[0300] 2-Chloro-5,7-dihydrodispiro[imidazo[1,2-e]purine-8,1'-cyclohexane-4',2''-[1,3]dioxolane] (5 g, 15.5 mmol) was dissolved in acetonitrile (70 mL), methyl iodide (66 g, 465 mmol) was added, and the mixture was stirred at 105 °C by microwave for 1 hour. The product was obtained by filtration (3.8 g, yield 73.1%).

[0301] 7. Preparation of 2'-chloro-5'-methyl-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purine]-4-one

[0302]

[0303] Dissolve 2-chloro-5-methyl-5,7-dihydrodispiro[imidazo[1,2-e]purine-8,1'-cyclohexane-4',2”- [1,3]dioxolane] (3.5 g, 10.4 mmol) in THF (76 mL), add concentrated hydrochloric acid (20 mL), stir at 16 °C for 6 hours, add it to an aqueous sodium carbonate solution to maintain the pH at 8 - 9, extract with EA to obtain the product (1.2 g, yield 40%).

[0304] 8. Preparation of 5'-methyl-2'-((7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)amino)-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purine]-4-one

[0305]

[0306] Dissolve 2'-chloro-5'-methyl-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purine]-4-one (50 mg, 0.17 mmol) in dioxane (3 mL), add 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (25.2 mg, 0.17 mmol), Brettphos Pd G3 (15.4 mg, 0.017 mmol), Cs2CO3 (110.8 mg, 0.34 mmol), stir at 100 °C for 6 hours under nitrogen protection. Rotavaporize the solvent and purify by silica gel column chromatography (DCM / MeOH = 40 / 1 - 15 / 1) to obtain the product (2.5 mg, yield 3.6%).

[0307] Molecular formula: C 20 H 21 N9O Molecular weight: 403.5 LC-MS (M / e): 404.2 (M + H + )

[0308] 1 H-NMR (400 MHz, CDCl3) δ: 9.03 (s, 1H), 8.54 (s, 1H), 8.36 (s, 1H), 7.81 (s, 1H), 7.66 (s, 1H), 4.14 (s, 2H), 3.39 (s, 3H), 2.63 - 2.58 (m, 2H), 2.50 - 2.45 (m, 2H), 2.34 (s, 3H), 2.14 - 2.01 (m, 2H), 1.99 - 1.91 (m, 2H).

[0309] Example 7: Preparation of 5'-Methyl-2'-((7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)amino)-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purin]-4-ol (Compounds 7-1 and 7-2)

[0310] 1. Preparation of 2'-Chloro-5'-methyl-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purin]-4-ol

[0311]

[0312] Dissolve 2'-Chloro-5'-methyl-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purin]-4-one (800.0 mg, 2.7 mmol) in MeOH (10.0 mL), add sodium borohydride (43.2 mg, 5.4 mmol), stir at 16 °C for 1 hour, rotary evaporate the solvent and purify by silica gel column chromatography (DCM / MeOH = 40 / 1 - 20 / 1) to obtain the product (350.0 mg, yield 43.5%).

[0313] 2. Preparation of 5'-Methyl-2'-((7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)amino)-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purin]-4-ol

[0314]

[0315] Dissolve 2'-Chloro-5'-methyl-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purin]-4-ol (59.0 mg, 0.2 mmol) in dioxane (3 mL), add 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (29.6 mg, 0.2 mmol), Brettphos Pd G3 (18.2 mg, 0.02 mmol), Cs2CO3 (130.3 mg, 0.4 mmol), stir at 100 °C for 2 hours under nitrogen protection. Purify by preparative TLC (DCM / MeOH = 8 / 1) to obtain Product 7-1 and Product 7-2.

[0316] Molecular formula: C 20 H 23 N9O Molecular weight: 405.5 LC-MS (M / e): 406.3 (M+H + )

[0317] The retention times, yields and 1H NMR spectra of the two products obtained are as follows:

[0318] 1) HPLC retention time RT: 6.6 min (high polarity); 14.0 mg, yield 17.2%;

[0319] 1 1H-NMR (400 MHz, CDCl3) δ: 10.01 (s, 1H), 8.24 (s, 1H), 7.60 (s, 1H), 7.55 (s, 1H), 6.73 (s, 1H), 4.15 (s, 2H), 3.51 (s, 3H), 2.85 - 2.78 (m, 2H), 2.78 (s, 3H), 2.11 - 2.03 (m, 2H), 1.77 - 1.66 (m, 4H).

[0320] 2) HPLC retention time RT: 6.7 min (low polarity); 15.6 mg, yield 19.2%;

[0321] 1 1H-NMR (400 MHz, CDCl3) δ: 9.77 (s, 1H), 8.27 (s, 1H), 7.58 (s, 1H), 7.28 (s, 1H), 6.60 (s, 1H), 4.18 (s, 2H), 3.93 (m, 1H), 3.88 (s, 1H), 4.18 (s, 3H), 2.43 (s, 3H), 2.26 - 2.10 (m, 2H), 2.02 - 1.95 (m, 2H), 1.90 - 1.85 (m, 2H), 1.52 - 1.49 (m, 4H).

[0322] Example 8: Preparation of 5-Methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-2',3',5,5',6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-thiopyran]-2-amine (Compound 8)

[0323] 1. Preparation of 4-Aminotetrahydro-2H-thiopyran-4-carbonitrile

[0324]

[0325] Add tetrahydro-4H-thiopyran-4-one (2.4 g, 20.7 mmol) to 7 M ammonia methanol solution (10 mL, 70.0 mmol), react at 0 °C for 4 hours, then add TMSCN (2.05 g, 20.7 mmol) at 0 °C, and then react at 20 °C for 4 hours. Then concentrate the system to obtain the crude product (2.9 g).

[0326] 2. Preparation of 4-((2-Chloro-5-nitropyrimidin-4-yl)amino)tetrahydro-2H-thiopyran-4-carbonitrile

[0327]

[0328] 2,4-Dichloro-5-nitropyrimidine (3.7 g, 19.3 mmol) and DIEA (5.0 g, 38.7 mmol) were added to THF (60 mL). 4-Aminotetrahydro-2H-thiopyran-4-carbonitrile (2.75 g of crude product) was added at -20 °C, and then the reaction was carried out at 25 °C for 4 hours. Then the system was concentrated and purified by silica gel column chromatography (PE:EA = 3:1) to obtain the target compound (4.0 g, yield 69.2%).

[0329] 3. Preparation of 4-((5-Amino-2-chloropyrimidin-4-yl)amino)tetrahydro-2H-thiopyran-4-carbonitrile

[0330]

[0331] 4-((2-Chloro-5-nitropyrimidin-4-yl)amino)tetrahydro-2H-thiopyran-4-carbonitrile (3.6 g, 12.0 mmol) and iron powder (2.7 g, 48.2 mmol) were added to a mixed system of HOAc (20 mL), ethanol (20 mL) and water (20 mL). Then the reaction was carried out at 75 °C for 1 hour. Then the system was cooled to 15 °C, and then filtered under reduced pressure. The filter cake was washed with (DCM:MeOH = 10:1) (20 mL). The obtained filtrate was concentrated by evaporation and purified by silica gel column chromatography (DCM:MeOH = 40:1) to obtain the product (1.3 g, yield 40.2%).

[0332] 4. Preparation of 4-(2-Chloro-8-thioxo-7,8-dihydro-9H-purin-9-yl)tetrahydro-2H-thiopyran-4-carbonitrile

[0333]

[0334] 4-((5-Amino-2-chloropyrimidin-4-yl)amino)tetrahydro-2H-thiopyran-4-carbonitrile (1.1 g, 4.1 mmol), imidazole (560 mg, 8.2 mmol) and S-CDI (1.45 g, 8.1 mmol) were added to DCM (30 mL). Then the system was reacted at 15 °C for 2 hours. Then the pH of the system was adjusted to 6 with 1N HCl. A solid precipitated from the system, and then it was filtered under reduced pressure. The filter cake was dried to obtain the target compound (900 mg, yield 70.4%).

[0335] 5. Preparation of 2-Chloro-2',3',5',6,6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-thiopyran]

[0336]

[0337] 4-(2-Chloro-8-thioxo-7,8-dihydro-9H-purin-9-yl)tetrahydro-2H-thiopyran-4-carbonitrile (800 mg, 2.6 mmol) was added to THF (30 mL), and then lithium aluminum hydride (293 mg, 7.7 mmol) was added portionwise at 40 °C. The mixture was then reacted at 40 °C for 1 h. Subsequently, the reaction system was cooled to 15 °C, and water (0.5 mL) was slowly added to quench the reaction. Then (DCM:MeOH = 10:1) (40 mL) was added and stirred for 0.5 h. After that, filtration was carried out under reduced pressure, and the filter cake was washed with (DCM:MeOH = 10:1) (50 mL). The obtained filtrate was concentrated and purified by silica gel column chromatography (DCM:MeOH = 35:1) to give the product (150 mg, yield 20.5%).

[0338] 6. Preparation of 2-Chloro-5-methyl-2',3',5,5',6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-thiopyran]

[0339]

[0340] 2-Chloro-2',3',5',6,6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-thiopyran] (140 mg, 0.50 mmol) was added to acetonitrile (5 mL), and then methyl iodide (1 mL) was added to the system. The mixture was then reacted under microwave at 105 °C for 1 h. Finally, the reaction system was concentrated, and saturated sodium bicarbonate solution (0.5 mL) was added. After purification by silica gel column chromatography (DCM:MeOH = 35:1), the product (85 mg, yield 57.5%) was obtained.

[0341] 7. Preparation of 5-Methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-2',3',5,5',6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-thiopyran]-2-amine

[0342]

[0343] 2-Chloro-5-methyl-2',3',5,5',6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-thiopyran] (75 mg, 0.25 mmol), 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (41 mg, 0.28 mmol), cesium carbonate (163 mg, 0.50 mmol) and BrettPhos Pd G3 (23 mg, 0.025 mmol) were dissolved in 1,4-dioxane (10 mL), and then the system was reacted at 105 °C for 4 h under N2 atmosphere. Then the system was concentrated and purified by silica gel column chromatography (DCM:MeOH = 20:1) to obtain the product (65 mg, yield 63.8%).

[0344] Molecular formula: C 19 H 21 N9S Molecular weight: 407.5 LC-MS (M / e): 408.2 (M+H + )

[0345] 1 1H NMR (400 MHz, CDCl3): δ 9.80 (s, 1H), 8.27 (s, 1H), 7.61 - 7.55 (m, 2H), 6.58 (s, 1H), 4.10 (s, 2H), 3.26 (s, 3H), 2.95 - 2.88 (m, 2H), 2.79 - 2.70 (m, 2H), 2.69 - 2.59 (m, 2H), 2.52 (s, 3H), 2.18 - 2.12 (m, 2H).

[0346] Example 9: Preparation of 5-methyl-2-((7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)amino)-2',3',5,5',6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-thiopyran] 1',1'-dioxide (Compound 9)

[0347] 1. Preparation of 2-chloro-5-methyl-2',3',5,5',6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-thiopyran] 1,1'-dioxide

[0348]

[0349] 2-Chloro-5-methyl-2',3',5',6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-thiopyran] (110 mg, 0.37 mmol) was added to DCM (10 mL), then mCPBA (80%, 240 mg, 1.11 mmol) was added to the system, and the reaction was carried out at 20 °C for 2 hours. The reaction was quenched by adding saturated sodium bicarbonate solution (5 mL), and then the mixture was extracted and separated. The organic phase was purified by preparative large plate (DCM:MeOH = 15:1) to obtain the product (25 mg, yield 20.6%).

[0350] 2. Preparation of 5-methyl-2-((7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)amino)-2',3',5,5',6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-thiopyran] 1',1'-dioxide

[0351]

[0352] 2-Chloro-5-methyl-2',3',5,5',6',7-hexahydrospiro[imidazo[1,2-e]purine-8,4'-thiopyran] 1',1'-dioxide (20 mg, 0.061 mmol), 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (11 mg, 0.074 mmol), cesium carbonate (40 mg, 0.12 mmol) and BrettPhos Pd G3 (6 mg, 0.0066 mmol) were dissolved in 1,4-dioxane (8 mL), and then the system was reacted at 110 °C for 6 hours under N2 atmosphere. Then the system was concentrated and purified by silica gel column chromatography (DCM:MeOH = 15:1) to obtain the product (6 mg, yield 22.4%).

[0353] Molecular formula: C 19 H 21 N9O2S Molecular weight: 439.5 LC-MS (M / e): 440.2 (M+H + )

[0354] 1 1H NMR (400 MHz, CDCl3): δ 9.65 (s, 1H), 8.29 (s, 1H), 7.76 - 7.51 (m, 2H), 6.68 (s, 1H), 4.17 (s, 2H), 4.12 - 4.02 (m, 2H), 3.51 (s, 3H), 3.16 - 3.08 (m, 2H), 2.66 - 2.58 (m, 2H), 2.56 - 2.45 (m, 5H).

[0355] Example 10: Preparation of 4-methoxy-4,5'-dimethyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purine]-2'-amine (Compound 15-1)

[0356] 1. Preparation of 8-methyl-1,4-dioxaspiro[4.5]decan-8-ol

[0357]

[0358] Dissolve 1,4-dioxaspiro[4.5]decan-8-one (25.0 g, 0.16 mol) in THF (300 mL). After the system is cooled to 0 °C, add a 3M ethereal solution of methylmagnesium bromide (100.0 mL, 0.30 mol). After the addition, warm the mixture to 25 °C and react for 5.0 hours. Quench the reaction by adding water, and purify the product by concentration and column chromatography (EA:PE = 50%) to obtain the target compound (13.0 g, yield 47.1%).

[0359] 2. Preparation of 8-methoxy-8-methyl-1,4-dioxaspiro[4.5]decane

[0360]

[0361] Dissolve 8-methyl-1,4-dioxaspiro[4.5]decan-8-ol (12.0 g, 69.7 mmol) in THF (100 mL). Add 60% NaH (14.0 g, 350.0 mmol) to the system. After the addition, react at 10 °C for 2 hours, then add CH3I (50.0 g, 352.0 mmol) and continue the reaction for 5.0 hours. Quench the reaction by adding water, and purify the product by concentration and column chromatography (EA:PE = 15%) to obtain the target compound (10.0 g, yield 76.9%).

[0362] 3. Preparation of 4-methoxy-4-methylcyclohexan-1-one

[0363]

[0364] Dissolve 8-methoxy-8-methyl-1,4-dioxaspiro[4.5]decane (10.0 g, 53.7 mmol) in THF (100 mL). Add 5M HCl solution at 25 °C and react for 2.0 hours. Then neutralize the system to pH 7 with sodium carbonate solution, and extract and concentrate and purify by column chromatography (EA:PE = 20%) to obtain the target compound (6.3 g, yield 82.9%).

[0365] 4. Preparation of 1-amino-4-methoxy-4-methylcyclohexane-1-carbonitrile

[0366]

[0367] Dissolve 4-methoxy-4-methylcyclohexan-1-one (6.3 g, 44.3 mmol) in 7M ammonia methanol solution (40.0 mL). After stirring at 25 °C for 3.0 h, trimethylsilyl cyanide (45.0 g, 45.4 mmol) was added dropwise. After continuing to stir for 5.0 h, it was directly concentrated for the next reaction.

[0368] 5. Preparation of 2'-chloro-4-methoxy-4,5'-dimethyl-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purine]

[0369]

[0370] Using 1-amino-4-methoxy-4-methylcyclohexane-1-carbonitrile as the starting material, referring to the preparation process of Steps 2 to 6 of Example 4 (Compound 4), 2'-chloro-4-methoxy-4,5'-dimethyl-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purine] was prepared.

[0371] 6. Preparation of 4-methoxy-4,5'-dimethyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purin]-2'-amine

[0372]

[0373] Dissolve 2'-chloro-4-methoxy-4,5'-dimethyl-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purine] (100.0 mg, 0.31 mmol), 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (51.0 mg, 0.34 mmol), Cs2CO3 (210.0 mg, 0.64 mmol) and BrettPhos Pd G3 (60.0 mg, 0.07 mmol) in dioxane (10.0 mL). The system was reacted at 100 °C under N2 for 10.0 h. After completion, it was concentrated and purified by column chromatography (DCM:MeOH = 10:1) to obtain the target product (13.0 mg, yield 9.7%).

[0374] Molecular formula: C 22 H 27 N9O Molecular weight: 433.5 LC-MS (M / e): 434.0 (M+H + )

[0375] 1H-NMR (400 MHz, DMSO-d6) δ: 9.71 (s, 1H), 8.26 (s, 1H), 7.5 (m, 2H), 6.61 (s, 1H), 4.11 (s, 2H), 3.45 (s, 3H), 3.35 (s, 3H), 2.73 - 2.62 (t, 2H), 2.51 (s, 3H), 2.11 - 1.921 (m, 2H), 1.53 - 1.42 (m, 2H), 1.25 (s, 3H), 0.91 - 0.81 (m, 2H).

[0376] HPLC retention time (RT): 3.879 min.

[0377] Example XI: Preparation of 4-methoxy-5'-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purine]-2'-amine (Compounds 16-1 and 16-2)

[0378] 1. Preparation of 2'-chloro-4-methoxy-5'-methyl-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purine]

[0379]

[0380] Dissolve 2'-chloro-5'-methyl-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purine]-4-ol (190.0 mg, 0.65 mmol) in THF (6.0 mL), add 60% NaH (78.0 mg, 1.95 mmol), stir at 16 °C for 15 minutes, add methyl iodide (306.5 mg, 2.2 mmol), stir at 60 °C for 1 hour, evaporate the solvent and purify by silica gel column chromatography (DCM / MeOH = 60 / 1 - 20 / 1) to obtain the product (90.0 mg, yield 45.2%).

[0381] 2. Preparation of 4-methoxy-5'-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purine]-2'-amine

[0382]

[0383] Dissolve 2'-chloro-4-methoxy-5'-methyl-5',7'-dihydrospiro[cyclohexane-1,8'-imidazo[1,2-e]purine] (70.0 mg, 0.23 mmol) in dioxane (4 mL), add 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (34.1 mg, 0.23 mmol), Brettphos Pd G3 (18.2 mg, 0.02 mmol), Cs2CO3 (150.0 mg, 0.46 mmol), and stir at 100 °C for 2 hours under nitrogen protection. Purify by preparative TLC (DCM / MeOH = 8 / 1) to obtain Product 16-1 and Product 16-2.

[0384] Molecular formula: C 21 H 25 N9O Molecular weight: 419.5 LC-MS (M / e): 420.3 (M+H + )

[0385] The retention times, yields, and 1H-NMR spectra of the two obtained products are as follows:

[0386] 1) HPLC retention time (RT): 9.35 min (more polar); 16.8 mg, yield 17.6%;

[0387] 1 1H-NMR (400 MHz, CDCl3) δ: 9.73 (s, 1H), 8.26 (s, 1H), 7.57 (s, 2H), 6.66 (s, 1H), 4.15 (s, 2H), 3.77 - 3.53 (m, 1H), 3.49 (m, 3H), 3.46 (s, 3H), 2.59 - 2.55 (m, 2H), 2.52 (s, 3H), 2.05 - 2.02 (m, 2H), 1.69 - 1.61 (m, 2H), 1.59 - 1.51 (m, 2H).

[0388] 2) HPLC retention time (RT): 9.49 min (less polar); 18.8 mg, yield 19.7%;

[0389] 1 1H-NMR (400 MHz, CDCl3) δ: 9.77 (s, 1H), 8.27 (s, 1H), 7.57 (s, 2H), 6.56 (s, 1H), 4.15 (s, 2H), 3.41 (s, 3H), 3.35 (s, 3H), 2.51 (s, 3H), 2.40 - 2.29 (m, 2H), 2.28 - 2.15 (m, 2H), 1.99 - 1.90 (m, 2H), 1.45 - 1.35 (m, 2H).

[0390] Example 12: Preparation of 2-(5,8-dimethyl-2-((7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)amino)-7,8-dihydro-5H-imidazo[1,2-e]purin-8-yl)ethan-1-ol (Compound 17)

[0391] 1. Preparation of 4-(benzyloxy)butan-2-one

[0392]

[0393] Dissolve 4-hydroxybutan-2-one (5.3 g, 60 mmol) in benzyl bromide (15.4 g, 90 mmol), add DIEA (15.8 mL, 96 mmol), heat to 150 °C and stir for 2 hours. Dilute with ethyl acetate, wash with 1 M hydrochloric acid, wash with saturated brine, dry over anhydrous sodium sulfate, filter and concentrate. Purify by column chromatography (ethyl acetate / petroleum ether = 0 - 40%) to obtain the product (10 g, yield 93%).

[0394] 2. Preparation of 2-amino-4-(benzyloxy)-2-methylbutanenitrile

[0395]

[0396] Dissolve 4-(benzyloxy)butan-2-one (10 g, 56 mmol) in a solution of ammonia in tetrahydrofuran (27 mL), stir at 0 °C for 4 hours, add TMSCN (5.6 g, 56 mmol), heat to 20 °C and stir for 4 hours. Evaporate to dryness to obtain the product directly for the next step of the reaction.

[0397] 3. Preparation of 4-(benzyloxy)-2-((2-chloro-5-nitropyrimidin-4-yl)amino)-2-methylbutanenitrile

[0398]

[0399] Dissolve 2,4-dichloro-5-nitropyrimidine (7.7 g, 40 mmol) in THF (100 mL), stir at 0 °C, add 2-amino-4-(benzyloxy)-2-methylbutanenitrile (crude product from the previous step), DIEA (10.3 g, 80 mmol), heat to 20 °C and continue stirring for 1 hour. Evaporate to dryness directly for the next step of the reaction.

[0400] 4. Preparation of 2-((5-amino-2-chloropyrimidin-4-yl)amino)-4-(benzyloxy)-2-methylbutanenitrile

[0401]

[0402] Dissolve 4-(benzyloxy)-2-((2-chloro-5-nitropyrimidin-4-yl)amino)-2-methylbutanenitrile (crude) in EtOH (50 mL), add iron powder (9 g, 160 mmol), glacial acetic acid (50 mL), and water (50 mL). Heat to 70 °C and stir for 1 hour. Filter, add water (50 mL), adjust the pH > 7 with aqueous NaOH solution, and extract with dichloromethane. The organic phase is concentrated by rotary evaporation and purified by silica gel column chromatography (ethyl acetate / petroleum ether = 50 - 90%), to obtain the product (3.3 g, overall yield of three steps 17.7%).

[0403] 5. Preparation of 4-(benzyloxy)-2-(2-chloro-8-thioxo-7,8-dihydro-9H-purin-9-yl)-2-methylbutanenitrile

[0404]

[0405] Dissolve 2-((5-amino-2-chloropyrimidin-4-yl)amino)-4-(benzyloxy)-2-methylbutanenitrile (3.3 g, 9.9 mmol) in DCM (50 mL), add imidazole (1 g, 14.9 mmol), S-CDI (1.9 g, 10.9 mmol), stir at 25 °C for 1 hour, and filter to obtain the product (540 mg, yield 15%).

[0406] 6. Preparation of 8-(2-(benzyloxy)ethyl)-2-chloro-8-methyl-7,8-dihydro-6H-imidazo[1,2-e]purine

[0407]

[0408] Dissolve 4-(benzyloxy)-2-(2-chloro-8-thioxo-7,8-dihydro-9H-purin-9-yl)-2-methylbutanenitrile (540 mg, 1.4 mmol) in THF (15 mL), add LiAlH4 (165 mg, 4.3 mmol), stir at 40 °C for 30 minutes, quench the reaction with water, concentrate by rotary evaporation and purify by silica gel column chromatography (DCM / MeOH = 20 / 1), to obtain the product (230 mg, yield 46%).

[0409] 7. Preparation of 8-(2-(benzyloxy)ethyl)-2-chloro-5,8-dimethyl-7,8-dihydro-5H-imidazo[1,2-e]purine

[0410]

[0411] 8-(2-(Benzyloxy)ethyl)-2-chloro-8-methyl-7,8-dihydro-6H-imidazo[1,2-e]purine (230 mg, 0.67 mmol) was dissolved in acetonitrile (5 mL), methyl iodide (1.9 g, 13.4 mmol) was added, and the mixture was stirred at 100 °C under microwave irradiation for 1 hour. After evaporation to dryness, the residue was purified by silica gel column chromatography (MeOH / DCM = 0 - 10%) to obtain the product (200 mg, yield 83%).

[0412] 8. Preparation of 8-(2-(Benzyloxy)ethyl)-5,8-dimethyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-7,8-dihydro-5H-imidazo[1,2-e]purin-2-amine

[0413]

[0414] 8-(2-(Benzyloxy)ethyl)-2-chloro-5,8-dimethyl-7,8-dihydro-5H-imidazo[1,2-e]purine (200 mg, 0.56 mmol) was dissolved in dioxane (10 mL), 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (100 mg, 0.67 mmol), Brettphos Pd G3 (153 mg, 0.17 mmol), and Cs2CO3 (438 mg, 1.3 mmol) were added, and the mixture was stirred at 110 °C under nitrogen protection for 24 hours. After evaporation of the solvent, the residue was purified by silica gel column chromatography (MeOH / DCM = 0 - 10%) to obtain the product (190 mg, yield 72%).

[0415] 9. 2-(5,8-Dimethyl-2-((7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)amino)-7,8-dihydro-5H-imidazo[1,2-e]purin-8-yl)ethan-1-ol

[0416]

[0417] 8-(2-(Benzyloxy)ethyl)-5,8-dimethyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-7,8-dihydro-5H-imidazo[1,2-e]purin-2-amine (60 mg, 0.13 mmol) was dissolved in dichloromethane (10 mL), 1 M BBr3 (390 μL, 0.39 mmol) was added, and the mixture was stirred at 20 °C for 1 hour. Methanol was added to quench the reaction, and the mixture was washed with saturated sodium bicarbonate, extracted with dichloromethane, dried over anhydrous sodium sulfate, and separated by TLC preparative plate (MeOH / DCM = 1 / 3) to obtain the product (13 mg, yield 26%).

[0418] Molecular formula: C 18H 21 N9O Molecular weight: 379.4 LC-MS (M / e): 380.2 (M+H + )

[0419] 1 H-NMR (400 MHz, MeOD) δ: 9.37 (s, 1H), 8.32 (s, 1H), 7.74 (s, 1H), 7.64 (s, 1H), 4.25 - 3.97 (m, 2H), 3.85 - 3.65 (m, 2H), 3.32 (s, 3H), 2.52 (s, 3H), 2.37 - 2.02 (m, 2H), 1.73 (s, 3H).

[0420] Example XIII: Preparation of 5'-Methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-4,5,5',7'-tetrahydro-2H-spiro[furan-3,8'-imidazo[1,2-e]purine]-2'-amine (Compound 18)

[0421]

[0422] Using dihydrofuran-3(2H)-one as the starting material, referring to the preparation process of Example IV (Compound 4), the target compound 5'-Methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-4,5,5',7'-tetrahydro-2H-spiro[furan-3,8'-imidazo[1,2-e]purine]-2'-amine was prepared.

[0423] Molecular formula: C 18 H 19 N9O Molecular weight: 377.2 LC-MS (M / e): 378.2 (M+H + )

[0424] 1 H-NMR (400 MHz, CDCl3) δ: 9.84 (s, 1H), 8.27 (s, 1H), 7.63 (s, 1H), 7.56 (s, 1H), 6.65 (s, 1H), 4.48 - 4.40 (m, 1H), 4.37 (s, 2H), 4.31 (d, 1H), 4.12 - 4.10 (m, 1H), 3.84 (d, 1H), 3.5 (s, 3H), 2.77 - 2.71 (m, 1H), 2.5 (s, 3H), 2.26 - 2.17 (m, 1H).

[0425] Example 14: Preparation of 5-Methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-5,7-dihydrospiro[imidazo[1,2-e]purine-8,4'-oxepane]-2-amine (Compound 19)

[0426] 1. Preparation of 4-Oxoparomomycin

[0427]

[0428] Dissolve tetrahydro-4H-pyran-4-one (1.0 g, 10.0 mmol) in methanol (10.0 mL). Add barium oxide (0.3 g, 1.96 mmol) and ethyl N-methyl-N-nitroso-carbamate (1.3 g, 9.8 mmol) at 0 °C. After raising the temperature to 25 °C, stir for 10 hours. Concentrate and perform column chromatography (EA / PE = 50%) to obtain the product (450 mg, yield 39.4%).

[0429] 2. Preparation of 4-Aminooxepane-4-carbonitrile

[0430]

[0431] Dissolve 4-oxoparomomycin (3.4 g, 29.8 mmol) in 7M ammonia / methanol solution (17.0 mL). React at 25 °C for 5 hours. Then add trimethylsilyl cyanide (3.0 g, 30.2 mmol) and stir for 10 hours. Concentrate and directly use for the next step reaction.

[0432] 3. Preparation of 4-((2-Chloro-5-nitropyrimidin-4-yl)amino)oxepane-4-carbonitrile

[0433]

[0434] Dissolve the crude product from the previous step of 4-aminooxepane-4-carbonitrile in THF (50 mL). Add DIEA (7.9 g, 61.2 mmol) and 2,4-dichloro-5-nitropyrimidine (5.8 g, 29.9 mmol). React at 25 °C for 2 hours. Filter by suction and dry to obtain the product (5.5 g, two-step yield 62.1%).

[0435] 4. Preparation of 4-((5-Amino-2-chloropyrimidin-4-yl)amino)oxepane-4-carbonitrile

[0436]

[0437] 4-((2-Chloro-5-nitropyrimidin-4-yl)amino)oxepane-4-carbonitrile (4.5 g, 15.1 mmol) and iron powder (4.5 g, 80.4 mmol) were dissolved in a mixed solvent of 40 mL each of acetic acid, ethanol, and water, and reacted at 80 °C for 2 hours. After suction filtration, the filter cake was washed with water and dried to obtain the product (3.8 g, yield 95%).

[0438] 5. Preparation of 4-(2-Chloro-8-thioxo-7,8-dihydro-9H-purin-9-yl)oxepane-4-carbonitrile

[0439]

[0440] 4-((5-Amino-2-chloropyrimidin-4-yl)amino)oxepane-4-carbonitrile (2.2 g, 8.22 mmol) and imidazole (1.1 g, 16.44 mmol) were dissolved in DCM (40 mL), S-CDI (2.9 g, 16.44 mmol) was added, and the reaction was carried out at 25 °C for 4 hours. After the reaction was completed as detected by LCMS, the pH was adjusted to 5 with dilute hydrochloric acid (2 M), and a solid precipitated. After suction filtration, the filter cake was washed with water and dried to obtain the product (1.2 g, yield 47.2%).

[0441] 6. Preparation of 2-Chloro-6,7-dihydrospiro[imidazo[1,2-e]purine-8,4'-oxepane]

[0442]

[0443] 4-(2-Chloro-8-thioxo-7,8-dihydro-9H-purin-9-yl)oxepane-4-carbonitrile (1.05 g, 3.40 mmol) was dissolved in THF (15 mL), and LiAlH4 (383 mg, 10.08 mmol) was added in batches at 40 °C. After continuing the reaction for 20 min, the reaction was completed as detected by LCMS, quenched with water (3 mL), suction filtered, the filter cake was washed with (MeOH:DCM = 1:10), the filtrate was concentrated by rotary evaporation, and separated by silica gel column chromatography (MeOH:DCM = 1:10) to obtain the product (450 mg, yield 47.3%).

[0444] 7. Preparation of 2-Chloro-5-methyl-5,7-dihydrospiro[imidazo[1,2-e]purine-8,4'-oxepane]

[0445]

[0446] Dissolve 2-chloro-6,7-dihydrospiro[imidazo[1,2-e]purine-8,4'-oxepane] (260 mg, 0.93 mmol) in acetonitrile (13 mL), add methyl iodide (1.5 mL, 24 mmol), react at 100 °C for 1 hour. After detecting the completion of the reaction by LCMS, quench the reaction with saturated NaHCO3. Rotate the reaction solution to dryness, and separate by silica gel column chromatography (MeOH:DCM = 1:10) to obtain the product (100 mg, yield 36.6%).

[0447] 8. Preparation of 5-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-5,7-dihydrospiro[imidazo[1,2-e]purine-8,4'-oxepane]-2-amine

[0448]

[0449] Dissolve 2-chloro-5-methyl-5,7-dihydrospiro[imidazo[1,2-e]purine-8,4'-oxepane] (100 mg, 0.34 mmol), 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (53 mg, 0.36 mmol), Brettphos Pd G3 (36 mg, 0.04 mmol) and cesium carbonate (280 mg, 0.86 mmol) in 1,4-dioxane (10 mL), under nitrogen protection, react at 100 °C for 2 hours. After detecting the completion of the reaction by LCMS, rotate the reaction solution to dryness, and separate by silica gel column chromatography (MeOH:DCM = 1:10) to obtain the crude product. Then add the mixed solvent of MeOH:hexane:DCM (2 mL:20 mL:2 mL) and sonicate to have insoluble substances. Filter the insoluble substances under reduced pressure and dry the filter cake to obtain the product (63.7 mg, yield 46.2%).

[0450] Molecular formula: C 20 H 23 N9O Molecular weight: 405.5 LC-MS (M / e): 406.2 (M+H + )

[0451] 1 1H-NMR (400 MHz, CDCl3) δ: 9.77 (s, 1H), 8.27 (s, 1H), 7.56 - 7.60 (m, 2H), 6.57 (s, 1H), 4.12 - 4.22 (m, 2H), 3.95 - 4.05 (m, 1H), 3.80 - 3.90 (m, 2H), 3.70 - 3.79 (m, 1H), 3.37 (s, 3H), 2.52 - 2.62 (m, 2H), 2.61 (s, 3H), 2.00 - 2.16 (m, 3H), 1.75 - 1.85 (m, 1H).

[0452] Chiral Isomer Resolution

[0453] Resolution Method: High Performance Liquid Chromatography (Chiral Column)

[0454]

[0455] Resolution Results:

[0456] Two compounds were obtained, and their retention times, ee values, and corresponding hydrogen spectra were as follows:

[0457]

[0458] Example XV: Preparation of 5-Methyl-2-((7-Methyl-[1,2,4]Triazolo[1,5-a]Pyridin-6-Yl)Amino)-2',3',5,5',6',7-Hexahydrospiro[Imidazo[1,2-e]Purine-8,4'-Thiopyran]1'-Oxide (Compound 20)

[0459] Prepared according to the method of Example IX, in which in the first-step oxidation reaction, the amount of mCPBA used was 1.2 equivalents.

[0460] The following compounds shown in the table were prepared using the same or similar methods as those in the above examples:

[0461]

[0462] Experimental Scheme

[0463] The following provides exemplary experimental schemes for some compounds of the present invention to show the advantageous activities and beneficial technical effects of the compounds of the present invention. However, it should be understood that the following experimental schemes are merely examples of the content of the present invention and not limitations on the scope of the present invention.

[0464] Experimental Example 1 In Vitro Enzymatic Activity of the Compounds of the Invention

[0465] Abbreviations

[0466] EDTA: Ethylenediaminetetraacetic Acid

[0467] DMSO: Dimethyl Sulfoxide

[0468] Tris: Tris(Hydroxymethyl)Aminomethane

[0469] Brij-35: Polyoxyethylene Lauryl Ether

[0470] DTT: Dithiothreitol

[0471] Test Substances: Compounds of the present invention, and their structural formulas and preparation methods are shown in the examples.

[0472] Experimental Reagents:

[0473] Name Brand ADP-Glo Kinase Assay Promege DNA-PK Promege

[0474] Experimental method:

[0475] 1. Prepare 1-fold kinase buffer

[0476] 1) 1-fold kinase buffer

[0477] 40 mM Tris, pH 7.5

[0478] 0.0055% Brij-35

[0479] 20 mM MgCl2

[0480] 0.05 mM DTT

[0481] 2. Compound preparation

[0482] 1) The initial concentration for compound detection is 1 μM, prepared into 100-fold concentration, i.e., 100 μM. Take 2 μl of 10 mM compound, add 198 μl of 100% DMSO to prepare a 100 μM compound solution. Add 100 μl of 100-fold compound to the second well on the 96-well plate, and add 60 μl of 100% DMSO to the other wells. Take 30 μl of the compound from the second well and add it to the third well, and perform 3-fold dilution successively downwards for a total of 10 concentrations.

[0483] 2) Transfer 100 μl of 100% DMSO and the highest concentration (400 nM) of the positive control wortmannin to two empty wells as the Max well and the Min well respectively.

[0484] 3) Use Echo to transfer 50 nl of the compound to the reaction wells of the 384-well plate.

[0485] 3. Prepare 2x kinase solution

[0486] 1) Prepare 2-fold DNA-PK kinase solution using 1-fold kinase buffer.

[0487] 2) Transfer 2.5 μl of the 2-fold enzyme solution to the reaction wells of the 384-well plate.

[0488] 3) Oscillate and mix well, then let it stand at room temperature.

[0489] 4. Prepare 2x substrate solution

[0490] 1) Prepare 2-fold substrate solution using 1-fold kinase buffer.

[0491] 2) Transfer 2.5 μl of the 2-fold substrate solution to the reaction wells of the 384-well plate to initiate the reaction.

[0492] 3) Oscillate and mix well.

[0493] 5. Kinase Reaction and Termination

[0494] 1) Cover the 384-well plate and incubate at 28 °C for 3 hours.

[0495] 2) Transfer 5 μl of ADP-Glo reagent and incubate at 28 °C for 2 hours.

[0496] 6. Detection of Reaction Results

[0497] 1) Transfer 10 μl of kinase detection reagent to the reaction wells of the 384-well plate to terminate the reaction.

[0498] 2) Let it stand at room temperature for 30 minutes.

[0499] 7. Data Reading

[0500] Read the sample values on Envision.

[0501] 8. Calculation of Inhibition Rate

[0502] 1) Copy the data from Envision.

[0503] 2) Convert it into inhibition rate data.

[0504] Percent inhibition = (max - conversion) / (max - min) * 100. Where max is the conversion rate of the DMSO control, min is the conversion rate of the no enzyme activity control, and conversion is the conversion rate at each concentration of the test compound.

[0505] 3) Import the data into MS Excel and perform curve fitting using XLFit excel add-in version 5.4.0.8.

[0506] Experimental Results:

[0507] Table 1 In vitro Enzymatic Activity Data of the Compounds of the Present Invention

[0508]

[0509]

[0510] Experimental Conclusions:

[0511] The compounds of the present invention have good inhibitory effects on the kinase activity of DNA-PK. Compounds 1-20 of the specific implementation all have low IC 50 values.

[0512] Experimental Example 2 Liver Microsome Metabolic Stability Experiment of the Compounds of the Invention in Different Species

[0513] Test article: The compound of the present invention, prepared in-house. The chemical name and preparation method are shown in the preparation examples of the compound.

[0514] Experimental materials:

[0515] Microsomes from cynomolgus monkey livers, with a microsomal protein concentration of 20 mg·mL -1 .

[0516] Microsomes from SD rats and CD-1 mice, with a microsomal protein concentration of 20 mg·mL for both -1 .

[0517] Microsomes from human livers, with a microsomal protein concentration of 20 mg·mL -1 .

[0518] The experimental initiating factor β-NADPH was purchased from Solarbio; the phosphate buffer solution (PBS) at pH 7.4 was prepared in-house.

[0519] Preparation of test article solution:

[0520] Weigh an appropriate amount of the test article powder precisely, dissolve it in an appropriate amount of dimethyl sulfoxide (DMSO) to 1 mM, and then dilute it 20-fold with methanol to a working solution of 50 μM.

[0521] Experimental method:

[0522] Table 2. Composition of the incubation system for the hepatic microsomal metabolic stability experiment

[0523]

[0524] Experimental operation steps:

[0525] (1) According to the ratio in Table 2 "Composition of the experimental incubation system" above, take 5.85 mL of 100 mM PBS, 0.585 mL of 20 mM MgCl2 solution, and 3.57 mL of H2O for each compound to prepare the incubation system mixed solution 1 (without microsomes, test article, and β-NADPH). The positive control drug verapamil was used in this experiment to prove the normal activity of hepatic microsomal enzymes.

[0526] (2) Take out the microsomes (20 mg protein / mL) from the -80°C refrigerator and pre-incubate them on a 37°C water bath thermostatic shaker for 3 min.

[0527] (3) Take 1.9 mL of the incubation system mixed solution 1 for each compound and each species, and add 56 μL of microsomes of different species to prepare the incubation system mixed solution 2 (without the test article and β-NADPH).

[0528] (4) Sample group (containing microsomes and β-NADPH): Take 616 μL of the incubation system mixed solution 2, add 14 μL of the test article working solution with a concentration of 50 μM, and add 70 μL of the β-NADPH working solution with a concentration of 10 mM. Mix well and duplicate the samples. The sampling time points are 0 min, 5 min, 10 min, 20 min, 30 min, and 60 min. This sample group is used to evaluate the metabolic stability of the compound mediated by β-NADPH.

[0529] (5) Control group (containing microsomes, without β-NADPH, using water instead of β-NADPH): Take 264 μL of the incubation system mixed solution 2, add 6 μL of the test article working solution with a concentration of 50 μM, and add 30 μL of water. Mix well and duplicate the samples. The sampling time points are 0 min and 60 min. This negative control group is used to evaluate whether there is non-β-NADPH-mediated metabolism of the compound in the liver microsome incubation system.

[0530] (6) At each predetermined time point, take 50 μL of the incubated sample from the incubation sample tube and add it to the termination sample tube (containing 300 μL of cold terminator, an acetonitrile solution containing 50 ng / mL of the internal standard tolbutamide), vortex to terminate the reaction.

[0531] (7) After vortexing for 10 min, centrifuge for 5 min (12000 rpm).

[0532] (8) Take 100 μL of the supernatant, add 100 μL of water, vortex to mix well, and perform LC-MS / MS injection analysis.

[0533] Data analysis:

[0534] Convert to the percentage of the remaining amount through the ratio of the peak area of the test article to the internal standard in the following formula.

[0535]

[0536] Experimental results:

[0537] The compound of the present invention has good stability in the liver microsomes of the test species.

[0538] Experimental Example 3: Pharmacokinetics experiment of the compound of the present invention in CD1 mice

[0539] Test article: The compound of the present invention, self-made. The chemical name and preparation method are shown in the preparation examples of each compound.

[0540] Test animals: CD1 mice, female, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., 6 mice / compound / dosage route.

[0541] Preparation of test article solution:

[0542] Preparation method of blank solvent (1): Weigh 28 g of HP-β-CD, add an appropriate amount of injection water to dissolve it, and then make up the volume to 100 mL with injection water. Vortex and mix evenly to obtain 28% HP-β-CD.

[0543] Preparation method of blank solvent (2): Weigh 20 g of HPC, slowly add it to 500 mL of stirred purified water, then add 1 mL of Tween 80, stir until it is clear and transparent, make up the volume to 1000 mL, and stir evenly to obtain 2% HPC + 0.1% Tween 80.

[0544] IV (intravenous injection) administration:

[0545] Weigh an appropriate amount of the compound of the present invention, add DMA (10% - 20%, v / v), vortex to dissolve it, then add PEG400 (5% - 10%, v / v) (the dosages of DMA and PEG400 can be appropriately increased or decreased according to the solubility of the specific compound), vortex and mix evenly. Finally, add blank solvent (1) (an appropriate amount, if the compound is insoluble in blank solvent (1), an acid solution or alkali solution with a certain pH can be used instead), vortex and mix evenly, keep it warm at 50 °C for 10 - 20 min to prepare a clear solution with an appropriate concentration (which can be 0.2 mg / mL, 1 mg / mL, etc.) as the IV administration solution of the test compound.

[0546] PO (gavage) administration:

[0547] Weigh an appropriate amount of the compound of the example of the present invention, place it in a tissue grinder, add blank solvent (2) (an appropriate amount), and grind it evenly at a speed of 1000 revolutions per minute to prepare a suspension liquid medicine with an appropriate concentration (which can be 1 mg / mL, 5 mg / mL, etc.) as the PO administration liquid medicine of the test compound.

[0548] Experimental method

[0549] The IV administration volume is 5 mL / kg. If the IV administration dose of the compound is 1 mg / kg, the administration concentration is 0.2 mg / mL; if the IV administration dose of the compound is 5 mg / kg, the administration concentration is 1 mg / mL.

[0550] The PO administration volume is 10 mL / kg. If the PO administration dose of the compound is 10 mg / kg, the administration concentration is 1 mg / mL; if the PO administration dose of the compound is 50 mg / kg, the administration concentration is 5 mg / mL.

[0551] Blood sampling time points: 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, 24 h after administration. The blood sampling is carried out in the following way as shown in the table below:

[0552]

[0553] About 50 μL of whole blood was collected from the inner canthus of the eye at each time point, placed in an anticoagulant tube containing EDTA-K2 anticoagulant, centrifuged at 8000 revolutions per minute for 6 minutes at 4 °C to obtain plasma samples, and the plasma was frozen at -80 °C in a refrigerator for later analysis.

[0554] Analysis of plasma samples

[0555] The protein precipitation method was used: 20 μL of plasma sample was taken, 200 μL of internal standard (acetonitrile solution containing 50 ng / mL of tolbutamide) was added, vortexed for 10 minutes, then centrifuged at 4000 revolutions per minute for 20 minutes, 100 μL of the supernatant was taken, and then 100 μL of water was added, vortexed and mixed for 3 minutes, and the drug concentration in the plasma was analyzed by LC-MS / MS.

[0556] Experimental results and conclusions

[0557] Through the drug concentration-time curve, pharmacokinetic parameters were calculated, such as the exposure amount AUC 0-t (indicating the area under the drug-time curve from 0 to t), clearance rate CL, elimination half-life T 1 / 2 、peak time T max 、peak concentration C max 、steady-state apparent volume of distribution V ss 、absolute bioavailability F%, etc. It can be seen from the test results that the compound of the present invention has good pharmacokinetic properties and has a high exposure amount and bioavailability.

Claims

1. A compound of formula (IIa) or a pharmaceutically acceptable salt thereof, Among them, X5 is selected from N(R 7 )); X6 is selected from N; X is selected from CH2, NH, O or S; R 1 selected from H, halogen, hydroxyl, amino, nitro, cyano, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, amino C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 alkoxy or amino C 1-6 alkoxy; R 2 and R 3 are each independently selected from C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, amino C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 alkoxy, amino C 1-6 alkoxy; or R 2 、R 3 and, together with the carbon atom to which it is attached, form a 3- to 8-membered cycloalkyl or 3- to 8-membered heterocyclic group optionally substituted by 1 to 2 Q2s; each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, C 1-6 alkyl, halo C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6 alkoxy or halo C 1-6 alkoxy; R 7 selected from H, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl or amino C 1-6 alkyl.

2. A compound of formula (IIb) or a pharmaceutically acceptable salt thereof, Among them, X5 is selected from N; X6 is selected from N(R 7 )); X is selected from CH2, NH, O or S; R 1 selected from H, halogen, hydroxy, amino, nitro, cyano, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, amino C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 alkoxy or amino C 1-6 alkoxy; R 2 、R 3 are each independently selected from C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, amino C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 alkoxy, amino C 1-6 alkoxy; or R 2 , R 3 and together with the carbon atom to which it is attached form a 3- to 8-membered cycloalkyl or 3- to 8-membered heterocyclic group optionally substituted by 1 to 2 Q2s; each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, C 1-6 alkyl, halo C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6 alkoxy or halo C 1-6 alkoxy; R 7 selected from H, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl or amino C 1-6 alkyl.

3. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-2, Among them, R 2 、R 3 and, together with the carbon atom to which it is attached, form the following group optionally substituted with 1 to 2 Q2 groups: Each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, C 1-6 alkyl, halo C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6 alkoxy or halo C 1-6 alkoxy.

4. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-2, wherein, R 1 selected from H, halogen, hydroxyl, amino, nitro, cyano, C 1-6 alkyl, halo-C 1-6 alkyl, hydroxy-C 1-6 alkyl, amino-C 1-6 alkyl, C 1-6 alkoxy, halo-C 1-6 alkoxy, hydroxy-C 1-6 alkoxy or amino-C 1-6 alkoxy; R 2 、R 3 are each independently selected from C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, amino C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 alkoxy or amino C 1-6 alkoxy; or R 2 , R 3 and the carbon atom to which it is attached together form the following group optionally substituted with 1-2 Q2s: Each Q2 is independently selected from halogen, hydroxyl, amino, nitro, cyano, carbonyl, C 1-6 alkyl, halo-C 1-6 alkyl, C 1-6 alkylamino, (C 1-6 alkyl)2amino, C 1-6 alkoxy or halo-C 1-6 alkoxy; R 7 selected from H, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl or amino C 1-6 alkyl.

5. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-2, wherein, R 1 selected from H, halogen, hydroxy, amino, nitro, cyano, methyl, ethyl, propyl, isopropyl, monofluoromethyl, difluoromethyl, trifluoromethyl, hydroxymethyl, aminomethyl, methoxy, ethoxy, propoxy, isopropoxy, monofluoromethoxy, difluoromethoxy or trifluoromethoxy; R 2 、R 3 are each independently selected from methyl, ethyl, propyl, isopropyl, trifluoromethyl, trifluoroethyl, trifluoropropyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, aminomethyl, aminoethyl, aminopropyl, methoxy, ethoxy, propoxy; or R 2 , R 3 and together with the carbon atom to which it is attached form the following group optionally substituted with 1-2 Q2s: each Q2 is independently selected from halogen, hydroxy, amino, nitro, cyano, carbonyl, methyl, ethyl, propyl, isopropyl, trifluoromethyl, methylamino, ethylamino, dimethylamino, diethylamino, methoxy, ethoxy or trifluoromethoxy; R 7 selected from H, methyl, ethyl, propyl, isopropyl, trifluoromethyl, hydroxymethyl or aminomethyl.

6. The compound or a pharmaceutically acceptable salt thereof according to claim 1 or 2, selected from the following compounds:

7. A pharmaceutical preparation comprising the compound according to any one of claims 1-6 or a pharmaceutically acceptable salt thereof, characterized in that, Comprising one or more pharmaceutically acceptable excipients, and the pharmaceutical preparation is any pharmaceutically acceptable dosage form.

8. A pharmaceutical composition comprising the compound according to any one of claims 1-6 or a pharmaceutically acceptable salt thereof, characterized in that, Comprising one or more second therapeutic active agents, and the second therapeutic active agents are selected from anticancer agents, including mitosis inhibitors, alkylating agents, antimetabolites, DNA intercalators, antitumor antibiotics, growth factor inhibitors, signal transduction inhibitors, cell cycle inhibitors, retinoid receptor regulators, proteasome inhibitors, topoisomerase inhibitors, biological response modifiers, hormonal drugs, angiogenesis inhibitors, cell growth inhibitors, targeting antibodies, HMG-CoA reductase inhibitors and protein prenyltransferase inhibitors.

9. Use of the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-6, the pharmaceutical preparation according to claim 7, or the pharmaceutical composition according to claim 8 in the preparation of a drug for preventing and / or treating benign tumors or cancers, wherein the cancers include carcinoma in situ and metastatic cancers.

10. Use of the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-6, the pharmaceutical preparation according to claim 7, or the pharmaceutical composition according to claim 8 in the preparation of a drug for preventing and / or treating benign tumors or cancers, wherein the drug is used in combination with radiotherapy and / or one or more anticancer agents, and the cancers include carcinoma in situ and metastatic cancers.

11. Use of the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-6, the pharmaceutical preparation according to claim 7, or the pharmaceutical composition according to claim 8 in the preparation of a drug for making cancer cells sensitive to anticancer agents and / or radiotherapy.

12. A kit, comprising: (a) An effective amount of one or more compounds or pharmaceutically acceptable salts thereof according to any one of claims 1-6, and (b) an effective amount of one or more anticancer agents.

13. An intermediate of formula (V), Among them, X1, X2, X4 are selected from N; X3 is CH; The dashed bond --- is a chemical bond or does not exist, and adjacent dashed bonds are not simultaneously chemical bonds; Y is selected from halogen, amino, hydroxy or mercapto; X5, X6, R 2 , R 3 , R 7 , Q2 are as described in any one of claims 1-6.

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