Pyrrolopyridine WEE1 inhibitor

Abstract

The invention provides a compound with a pyrrolopyridine structure, which has WEE1 inhibitory activity and can be used for treating related diseases caused by WEE1 overexpression or hyperactivity, such as cancer.

Description

[0001] This application claims priority to a prior application with application number 202410285477.2, filed with the State Intellectual Property Office on March 13, 2024, and entitled “A Pyrrolopyridine WEE1 Inhibitor”. The full text of the prior application is incorporated into this application. Technical Field

[0002] The present application relates to the field of pharmaceutical compounds, and specifically to a pyrrolopyridine compound having WEE1 kinase target inhibitory activity, a pharmaceutical composition comprising the compound, and the use of the compound in the preparation of a drug for treating or preventing diseases associated with WEE1 overexpression or hyperactivity. Background Art

[0003] The cell cycle is closely related to the DNA damage repair process. The cell cycle refers to the entire process of cell division, which is divided into two stages: interphase and mitotic phase (M). The cell cycle checkpoint is a key point in regulating the cell cycle. Its main function is to ensure that each step in the cycle is highly regulated and controlled, and to adjust the cell state to adapt to the external environment. Eukaryotic cells rely on complex cell cycle checkpoints and DNA repair systems to ensure the stability of the genome. After the cell DNA is damaged by chemical or radioactive damage, the cell cycle checkpoints are activated, blocking the cell cycle process and allowing DNA to be repaired. The normal cell cycle has checkpoints in the G1 / S phase, S phase and G2 / M phase, so that the DNA has enough time to repair damage. Under normal circumstances, DNA damage in cells can be repaired through two pathways mediated by the tumor suppressor gene p53: ATM / ATR-P53-CDK4 / CyclinD and ATM / ATR-P53-CDK2 / CyclinE. This inhibits the phosphorylation of the tumor suppressor gene Rb, arresting cells in the G1 phase and allowing DNA repair to complete. However, in tumor cells, the mutation rate of the tumor suppressor gene p53 is high, with mutations in over 50% of tumor cells. Tumor cells with p53 deficiency have inactivated the G1 / S checkpoint, leaving DNA repair primarily dependent on the G2 / M checkpoint.

[0004] WEE1 kinase, first discovered in fission yeast, is a key member of the serine / threonine protein kinase family. It is a cell cycle regulator that regulates the phosphorylation state of cyclin-dependent kinase 1 (CDK1), thereby modulating the activity of the CDK1-cyclin B complex and cell cycle regulation. It plays a crucial role in the G2 / M checkpoint of the cell cycle. WEE1 kinase inhibits CDK kinase activity by phosphorylating CDK at Tyr14 and Tyr15, arresting cells in the G2 / M phase of the cell cycle and allowing them ample time for DNA damage repair. Inhibition of WEE1 kinase further inactivates the G2 / M checkpoint, abolishing cell cycle arrest and driving cells into premature division, leading to mitotic catastrophe. This leads to synthetic lethality and induction of apoptosis in tumor cells (S. Mueller, J. Clin. Oncol., 2015). However, the p53 gene in normal cells functions normally, checking DNA integrity at the G1 / S and G2 / M checkpoints. When WEE1 kinase activity is inhibited, this functional loss is compensated by p53-dependent DNA damage repair mechanisms. In theory, inhibiting WEE1 kinase activity could selectively kill p53-deficient tumor cells while leaving normal cells unaffected, making it an ideal approach for tumor treatment.

[0005] Related studies have shown that WEE1 kinase is overexpressed in breast cancer, ovarian cancer, liver cancer, cervical cancer, lung cancer, squamous cell carcinoma, colorectal cancer, gastric cancer, glioblastoma, diffuse intramural glioma, and malignant melanoma. High expression in ovarian cancer, melanoma, and glioblastoma is associated with poor prognosis. Therefore, there is a need in this field to develop new WEE1 kinase inhibitors with good anticancer activity and high safety. Summary of the Invention

[0006] The present application discloses a compound of formula I, a pharmaceutically acceptable salt, solvate, or stereoisomer thereof,

[0007]

[0008] Wherein, n is an integer from 0 to 5;

[0009] R1 is independently selected from halogen, -OH, -CN, -NH2, -C1-8 alkyl, -C2-8 alkenyl, -C2-8 alkynyl, -C1-8 alkoxy, -C2-8 alkenyloxy, -C2-8 alkynyloxy, and the alkyl, alkenyl, alkynyl, alkoxy, alkenyloxy and alkynyloxy groups are optionally substituted with one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C1-4 alkoxy;

[0010] R2 is selected from H, halogen, -OH, -CN, -NH2, -C1-8 alkyl, -C2-8 alkenyl, -C2-8 alkynyl, -OR7, -N(R7)R8, wherein the alkyl, alkenyl, and alkynyl groups are optionally substituted with one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, -C1-4 alkoxy, C3-10 cycloalkyl, 3-10 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, and S, C6-10 aryl, and 5-11 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S; R7 is selected from -C1-8 alkyl, -C2-8 alkenyl, -C2-8 alkynyl, -(CH2) m -C3-10 cycloalkyl, -(CH2) p -3-10 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, S, -(CH2) r -C6-10 aryl, -(CH2) q - a 5-11 membered heteroaryl group containing 1-3 ring heteroatoms independently selected from N, O, and S, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are optionally substituted with one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, and -C1-4 alkoxy; R8 is selected from H, -C1-8 alkyl, -C2-8 alkenyl, and -C2-8 alkynyl, wherein the alkyl, alkenyl, and alkynyl groups are optionally substituted with one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, and -C1-4 alkoxy; m, p, r, and q are independently integers of 0 to 4;

[0011] R3, R4, R6 are independently selected from H, halogen, -OH, -CN, -NH2, -C1-8 alkyl, -C2-8 alkenyl, -C2-8 alkynyl, -C1-8 alkoxy;

[0012] R5 is selected from C3-10 cycloalkyl, 3-10 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, and S, C6-10 aryl, and 5-11 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S, wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted by one or more independently selected from =O, halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, -OR9, -C(O)R9, -N(R9)R 10wherein the alkyl, alkenyl and alkynyl groups are optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl and -C1-4 alkoxy; R9 is selected from -C1-8 alkyl, -C2-8 alkenyl and -C2-8 alkynyl, wherein the alkyl, alkenyl and alkynyl groups are optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl and -C1-4 alkoxy; R 10 Selected from H, -C1-8 alkyl, -C2-8 alkenyl, -C2-8 alkynyl, and the alkyl, alkenyl, and alkynyl are optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, and -C1-4 alkoxy.

[0013] In some preferred embodiments of the present invention, in Formula I, n is an integer from 0 to 5;

[0014] R1 is independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, -C1-4 alkoxy, -C2-4 alkenyloxy, -C2-4 alkynyloxy, and the alkyl, alkenyl, alkynyl, alkoxy, alkenyloxy and alkynyloxy groups are optionally substituted with one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C1-4 alkoxy;

[0015] R2 is selected from H, halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, -OR7, -N(R7)R8, wherein the alkyl, alkenyl, and alkynyl groups are optionally substituted with one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, -C1-4 alkoxy, C3-7 cycloalkyl, 3-7 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, and S, C6-10 aryl, and 5-11 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S; R7 is selected from -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, -(CH2) m -C3-7 cycloalkyl, -(CH2) p -3-7 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, S, -(CH2) r -C6-10 aryl, -(CH2) q-5-11 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are optionally substituted with one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, and -C1-4 alkoxy; R8 is selected from H, -C1-4 alkyl, -C2-4 alkenyl, and -C2-4 alkynyl, wherein the alkyl, alkenyl, and alkynyl groups are optionally substituted with one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, and -C1-4 alkoxy; m, p, r, and q are independently integers of 0 to 4;

[0016] R3, R4, R6 are independently selected from H, halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, -C1-4 alkoxy;

[0017] R5 is selected from C3-7 cycloalkyl, 3-7 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, and S, C6-10 aryl, and 5-11 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S, wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted by one or more independently selected from =O, halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, -OR9, -C(O)R9, -N(R9)R 10 wherein the alkyl, alkenyl and alkynyl groups are optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl and -C1-4 alkoxy; R9 is selected from -C1-4 alkyl, -C2-4 alkenyl and -C2-4 alkynyl, wherein the alkyl, alkenyl and alkynyl groups are optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl and -C1-4 alkoxy; R 10 Selected from H, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, and the alkyl, alkenyl, and alkynyl are optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, and -C1-4 alkoxy.

[0018] In some preferred embodiments of the present invention, n is an integer from 0 to 2.

[0019] In some preferred embodiments of the present invention, R1 is independently selected from F, Cl, Br, -OH, -CN, -NH2, -C1-4 alkyl, -C1-4 alkoxy, and the alkyl and alkoxy groups are optionally substituted by one or more groups independently selected from F, Cl, Br, -OH, -CN, -NH2, -C1-4 alkyl, -C1-4 alkoxy.

[0020] In some preferred embodiments of the present invention, R2 is selected from halogen, -C1-4 alkyl, -OR7, -N(R7)R8, and the alkyl group is optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C1-4 alkoxy, C3-7 cycloalkyl, 3-7 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, and S, C6-10 aryl, and 5-6 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S; R7 is selected from -C1-4 alkyl, -(CH2) m -C3-7 cycloalkyl, -(CH2) p -3-7 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, S, -(CH2) r -C6-10 aryl, -(CH2) q - a 5-6 membered heteroaryl group containing 1-3 ring heteroatoms independently selected from N, O, and S, wherein the alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are optionally substituted by one or more groups independently selected from -OH, -C1-4 alkyl, and -C1-4 alkoxy; R8 is selected from H, -C1-4 alkyl, and the alkyl group is optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, and -C1-4 alkoxy; m, p, r, and q are independently integers of 0 to 2.

[0021] In some preferred embodiments of the present invention, R3, R4, and R6 are independently selected from H, halogen, -C1-4 alkyl, and -C1-4 alkoxy.

[0022] In some preferred embodiments of the present invention, R5 is selected from C3-7 cycloalkyl, 3-7 membered heterocycloalkyl containing 1 ring heteroatom of N or O, C6-10 aryl, 5-6 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, S, and the cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted by one or more independently selected from =O, -C1-4 alkyl, -OR9, -C(O)R9, -N(R9)R 10wherein the alkyl group is optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C1-4 alkoxy; R9 is selected from -C1-4 alkyl, wherein the alkyl group is optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C1-4 alkoxy; R 10 Selected from -C1-4 alkyl, wherein the alkyl is optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C1-4 alkoxy.

[0023] In some embodiments of the present invention, the compound of formula I comprises one or more characteristics selected from the group consisting of:

[0024] (1) n is an integer from 0 to 2;

[0025] (2) R1 is independently selected from F, Cl, Br, -OH, -CN, -NH2, -C1-4 alkyl, -C1-4 alkoxy, and the alkyl and alkoxy groups are optionally substituted with one or more groups independently selected from F, Cl, Br, -OH, -CN, -NH2, -C1-4 alkyl, and -C1-4 alkoxy;

[0026] (3) R2 is selected from halogen, -C1-4 alkyl, -OR7, -N(R7)R8, wherein the alkyl group is optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C1-4 alkoxy, C3-7 cycloalkyl, 3-7 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, and S, C6-10 aryl, and 5-6 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S; R7 is selected from -C1-4 alkyl, -(CH2) m -C3-7 cycloalkyl, -(CH2) p -3-7 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, S, -(CH2) r -C6-10 aryl, -(CH2) q - a 5-6 membered heteroaryl group containing 1-3 ring heteroatoms independently selected from N, O, and S, wherein the alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are optionally substituted with one or more groups independently selected from -OH, -C1-4 alkyl, and -C1-4 alkoxy; R8 is selected from H, -C1-4 alkyl, and the alkyl group is optionally substituted with one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, and -C1-4 alkoxy; m, p, r, and q are independently integers of 0 to 2;

[0027] (4) R3, R4, and R6 are independently selected from H, halogen, -C1-4 alkyl, and -C1-4 alkoxy;

[0028] (5) R5 is selected from C3-7 cycloalkyl, 3-7 membered heterocycloalkyl containing 1 ring heteroatom of N or O, C6-10 aryl, 5-6 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, S, wherein the cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted by one or more independently selected from =O, -C1-4 alkyl, -OR9, -C(O)R9, -N(R9)R 10 wherein the alkyl group is optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C1-4 alkoxy; R9 is selected from -C1-4 alkyl, wherein the alkyl group is optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C1-4 alkoxy; R 10 Selected from -C1-4 alkyl, wherein the alkyl is optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C1-4 alkoxy.

[0029] In some preferred embodiments of the present invention, Formula I is as shown in Formula I-1,

[0030] in:

[0031] Each R1 is independently selected from F, Cl, Br, CH3, CH2CH3, CH3O;

[0032] R2 is selected from halogen, -C1-4 alkyl, -OR7, -N(R7)R8, wherein the alkyl group is optionally substituted by one or more groups independently selected from halogen, -C1-4 alkyl, -C1-4 alkoxy, C3-7 cycloalkyl, 3-7 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, and S, C6-10 aryl, and 5-6 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S; R7 is selected from -C1-4 alkyl, -(CH2) m -C3-7 cycloalkyl, -(CH2) p -3-7 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, S, -(CH2) r -C6-10 aryl, -(CH2) q- a 5-6 membered heteroaryl group containing 1-3 ring heteroatoms independently selected from N, O, and S, wherein the alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are optionally substituted with one or more groups independently selected from halogen, -OH, -C1-4 alkyl, and -C1-4 alkoxy; R8 is selected from H, -C1-4 alkyl; m, p, r, and q are independently integers of 0 to 1;

[0033] R3, R4, R6 are independently selected from H, halogen, -C1-4 alkyl, -C1-4 alkoxy;

[0034] R5 is selected from C3-7 cycloalkyl, 4-6 membered heterocycloalkyl containing 1 ring heteroatom selected from N or O, 5-6 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, S, and the cycloalkyl, heterocycloalkyl and heteroaryl are optionally substituted by one or more independently selected from =O, -C1-4 alkyl, -OR9, -C(O)R9, -N(R9)R 10 wherein the alkyl group is optionally substituted by one or more groups independently selected from halogen, -C1-4 alkyl, -C1-4 alkoxy; R9 is selected from -C1-4 alkyl; R 10 Selected from -C1-4 alkyl.

[0035] In some preferred embodiments of the present invention, in formula I-1, R3, R4, and R6 are H.

[0036] In some preferred embodiments of the present invention, in formula I-1, R2 is selected from F, Cl, Br, -C1-4 alkyl, -OR7, -N(R7)R8, and the alkyl group is optionally substituted by one or more groups independently selected from halogen, -C1-4 alkyl, -C1-4 alkoxy, C3-7 cycloalkyl, 3-7 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, and S, C6-10 aryl, and 5-6 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S; R7 is selected from -C1-4 alkyl, -C3-7 cycloalkyl, -(CH2) q - a 5-6 membered heteroaryl group containing 1-3 ring heteroatoms independently selected from N, O, and S, wherein the cycloalkyl group is optionally substituted by a group selected from -C1-4 alkoxy, and the alkyl group is optionally substituted by one or more groups independently selected from F, Cl, and Br; R8 is selected from H, -C1-4 alkyl; and q is an integer from 0 to 1. Preferably, R2 is selected from F, Cl, Br, -C1-4 alkyl, -OR7, or -N(R7)R8, wherein the alkyl group is optionally substituted by one or more groups independently selected from F, Cl, Br, or -C1-4 alkoxy; R7 is selected from -C1-4 alkyl, -C3-7 cycloalkyl, -(CH2) q- a 5-6 membered heteroaryl group containing 1-3 ring heteroatoms independently selected from N, O, and S, wherein the cycloalkyl group is optionally substituted by a group selected from -C1-4 alkoxy, and the alkyl group is optionally substituted by one or more groups independently selected from F, Cl, and Br; R8 is selected from H, -C1-4 alkyl; q is an integer from 0 to 1.

[0037] In some preferred embodiments of the present invention, in formula I-1, R5 is selected from C3-7 cycloalkyl, 4-6 membered heterocycloalkyl containing 1 ring heteroatom selected from N or O, 5-6 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, S, and the cycloalkyl is replaced by a -N(R9)R 10 The heterocycloalkyl group is connected to the pyrazole group through a ring carbon atom, and the heterocycloalkyl group is optionally substituted by one or more groups independently selected from =O, -C1-4 alkyl, -OR9, and -C(O)R9, wherein the alkyl group is optionally substituted by one or more groups independently selected from F, Cl, Br, -C1-4 alkyl, and -C1-4 alkoxy; R9 is selected from -C1-4 alkyl; R 10 Preferably, R5 is selected from C4-6 cycloalkyl, 4-6 membered heterocycloalkyl containing one ring heteroatom selected from N or O, said cycloalkyl being separated by a -N(R9)R 10 wherein the heterocycloalkyl group is connected to the pyrazole group via a ring carbon atom, and the heterocycloalkyl group is optionally substituted by one or more groups independently selected from =O, -C1-4 alkyl, and -C(O)R9. When the heterocycloalkyl group is an N-containing heterocycloalkyl group, if it has a -C1-4 alkyl or -C(O)R9 substituent, the substituent is located on the ring N atom, wherein the alkyl group is optionally substituted by one or more groups independently selected from F, Cl, Br, and -C1-4 alkoxy; R9 is selected from -C1-4 alkyl; R 10 Selected from -C1-4 alkyl.

[0038] In some preferred embodiments of the present invention, the compound of formula I-1 comprises one or more characteristics selected from the group consisting of:

[0039] (i) R3, R4, and R6 are H;

[0040] (ii) R2 is selected from F, Cl, Br, -C1-4 alkyl, -OR7, -N(R7)R8, wherein the alkyl group is optionally substituted by one or more groups independently selected from halogen, -C1-4 alkyl, -C1-4 alkoxy, C3-7 cycloalkyl, 3-7 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, and S, -C6-10 aryl, or 5-6 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S; R7 is selected from -C1-4 alkyl, -C3-7 cycloalkyl, -(CH2)q -5-6 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S, wherein the cycloalkyl group is optionally substituted by a group selected from -C1-4 alkoxy, and the alkyl group is optionally substituted by one or more groups independently selected from F, Cl, and Br; R8 is selected from H, -C1-4 alkyl; q is an integer from 0 to 1;

[0041] (iii) R5 is selected from C3-7 cycloalkyl, 4-6 membered heterocycloalkyl containing 1 ring heteroatom selected from N or O, 5-6 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, S, wherein the cycloalkyl is substituted by a -N(R9)R 10 The heterocycloalkyl group is connected to the pyrazole group through a ring carbon atom, and the heterocycloalkyl group is optionally substituted by one or more groups independently selected from =O, -C1-4 alkyl, -OR9, and -C(O)R9, wherein the alkyl group is optionally substituted by one or more groups independently selected from F, Cl, Br, -C1-4 alkyl, and -C1-4 alkoxy; R9 is selected from -C1-4 alkyl; R 10 Selected from -C1-4 alkyl.

[0042] In some further embodiments of the present invention, in Formula I-1:

[0043] R3, R4, and R6 are H;

[0044] R2 is selected from F, Cl, Br, -C1-4 alkyl, -OR7, -N(R7)R8, wherein the alkyl group is optionally substituted by one or more groups independently selected from halogen, -C1-4 alkyl, -C1-4 alkoxy; R7 is selected from -C1-4 alkyl, -C3-7 cycloalkyl, -(CH2) q -5-6 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S, wherein the cycloalkyl group is optionally substituted by a group selected from -C1-4 alkoxy, and the alkyl group is optionally substituted by one or more groups independently selected from F, Cl, and Br; R8 is selected from H, -C1-4 alkyl; q is an integer from 0 to 1;

[0045] R5 is selected from C4-6 cycloalkyl, 4-6 membered heterocycloalkyl containing one ring heteroatom selected from N or O, said cycloalkyl being substituted by a -N(R9)R 10wherein the heterocycloalkyl group is connected to the pyrazole group via a ring carbon atom, and the heterocycloalkyl group is optionally substituted by one or more groups independently selected from =O, -C1-4 alkyl, and -C(O)R9. When the heterocycloalkyl group is an N-containing heterocycloalkyl group, if it has a -C1-4 alkyl or -C(O)R9 substituent, the substituent is located on the ring N atom, wherein the alkyl group is optionally substituted by one or more groups independently selected from F, Cl, Br, and -C1-4 alkoxy; R9 is selected from -C1-4 alkyl; R 10 Selected from -C1-4 alkyl.

[0046] In some further embodiments of the present invention, in Formula I-1:

[0047] R3, R4, and R6 are H;

[0048] R2 is selected from -OR7, -N(R7)R8; R7 is selected from -C1-4 alkyl, -C3-7 cycloalkyl, -(CH2) q -5-6 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S, wherein the alkyl group is optionally substituted by one or more groups independently selected from F, Cl, and Br, and the cycloalkyl group is optionally substituted by a group selected from -C1-4 alkoxy; R8 is selected from H, -C1-4 alkyl; q is an integer from 0 to 1;

[0049] R5 is selected from C4-6 cycloalkyl (preferably cyclobutyl, cyclopentyl, cyclohexyl), 5-6 membered heterocycloalkyl containing 1 O ring heteroatom (preferably ), a 4-6 membered heterocycloalkyl containing one N ring heteroatom (preferably ), the cycloalkyl group is replaced by a -N(R9)R 10 The heterocycloalkyl group is connected to the pyrazole group through a ring carbon atom, the ring N heteroatom of the heterocycloalkyl group is optionally substituted by a -C1-4 alkyl group, and the alkyl group is optionally substituted by one or more groups independently selected from F, Cl, Br, and -C1-4 alkoxy; R9 is selected from -C1-4 alkyl; R 10 Selected from -C1-4 alkyl.

[0050] In some further embodiments of the present invention, in Formula I-1:

[0051] R3, R4, and R6 are H;

[0052] R2 is selected from -OR7, -N(R7)R8, R7 is selected from -C1-4 alkyl, -C3-7 cycloalkyl, -(CH2) q- a 5-6 membered heteroaryl group containing 1-3 ring heteroatoms independently selected from N, O, and S, wherein the cycloalkyl group is optionally substituted by a group selected from -C1-4 alkoxy; R8 is selected from H, -C1-4 alkyl; q is an integer from 0 to 1;

[0053] R5 is selected from C4-6 cycloalkyl (preferably cyclobutyl, cyclopentyl, cyclohexyl), 4-6 membered heterocycloalkyl containing 1 N ring heteroatom (preferably ), the cycloalkyl group is replaced by a -N(R9)R 10 The heterocycloalkyl group is connected to the pyrazole group through a ring carbon atom, and the ring N heteroatom of the heterocycloalkyl group is optionally substituted by a -C1-4 alkyl group; R9 is selected from -C1-4 alkyl; R 10 Selected from -C1-4 alkyl.

[0054] In some embodiments of the present invention, the compound of formula I is selected from the following compounds:

[0055]

[0056]

[0057] The present invention also discloses a method for preparing a compound of formula I, comprising reacting a compound of formula A with a compound of formula B, and then removing the group SEM and / or further modifying the group as needed to form a compound of formula I, wherein R1-R6 and n are defined as above, SEM is 2-(trimethylsilyl)ethoxy)methyl, and X is a leaving group, such as a halogen, more preferably iodine.

[0058]

[0059] The compound of formula I of the present invention or its pharmaceutically acceptable salt, solvate, or stereoisomer has WEE1 inhibitory activity and can be used for the treatment of related diseases mediated by WEE1 overexpression or hyperactivity.

[0060] The present invention provides an in vitro non-therapeutic and non-diagnostic method for inhibiting WEE1, comprising the steps of contacting WEE1 or cells expressing WEE1 with the compound of the present invention or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof.

[0061] The present invention provides a method for treating diseases mediated by WEE1 overexpression or hyperactivity, comprising administering a therapeutically effective amount of the compound of the present invention or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof to a patient in need.

[0062] The present invention also provides use of the compound of the present invention or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof in the preparation of a drug for treating diseases mediated by overexpression or hyperactivity of WEE1.

[0063] Related diseases mediated by WEE1 overexpression or hyperactivity are, for example, tumors, including but not limited to: breast cancer, ovarian cancer, liver cancer, cervical cancer, lung cancer, squamous cell carcinoma, colorectal cancer, gastric cancer, glioblastoma, diffuse intramural glioma, melanoma, etc.

[0064] The inhibitory activity of the compounds of the present invention against WEE1 is determined by measuring the activity and phosphorylation level of pCDK1. Preferred compounds of the present invention have a Ki of ≤ 10 μM for pCDK1 phosphorylation, and more preferred compounds have a Ki of ≤ 1 μM. Furthermore, preferred compounds of the present invention have low inhibitory activity against PLK1, preferably, a Ki of ≥ 30 μM for PLK1.

[0065] The present invention also provides a pharmaceutical composition characterized by comprising a compound of formula I or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof.

[0066] The pharmaceutical composition further contains a pharmaceutically acceptable carrier.

[0067] The pharmaceutically acceptable carriers are various excipients commonly used or known in the pharmaceutical field, including but not limited to: diluents, binders, antioxidants, pH regulators, preservatives, lubricants, disintegrants, etc.

[0068] Examples of diluents include lactose, starch, cellulose derivatives, inorganic calcium salts, and sorbitol. Examples of binders include starch, gelatin, sodium carboxymethyl cellulose, and polyvinyl pyrrolidone. Examples of antioxidants include vitamin E, sodium bisulfite, sodium sulfite, and butylated hydroxyanisole. Examples of pH adjusters include hydrochloric acid, sodium hydroxide, citric acid, tartaric acid, Tris, acetic acid, sodium dihydrogen phosphate, and disodium hydrogen phosphate. Examples of preservatives include methyl parahydroxybenzoate, ethyl parahydroxybenzoate, m-cresol, and benzalkonium chloride. Examples of lubricants include magnesium stearate, micropowdered silica gel, and talc. Examples of disintegrants include starch, methyl cellulose, xanthan gum, and cross-linked sodium carboxymethyl cellulose.

[0069] The pharmaceutical composition may be in the form of an oral dosage form, such as a tablet, capsule, pill, powder, granule, suspension, syrup, etc.; or in the form of an injectable dosage form, such as an injection solution, powder injection, etc., which is administered by intravenous, intraperitoneal, subcutaneous, or intramuscular injection. All dosage forms are well known to those skilled in the pharmaceutical field.

[0070] Routes of administration of the pharmaceutical composition include, but are not limited to: oral; buccal; sublingual; transdermal; pulmonary; rectal; parenteral, e.g., by injection, including subcutaneous, intradermal, intramuscular, intravenous; by implanted reservoir or reservoir.

[0071] In the present invention, although substituents are disclosed as groups or ranges, the groups or ranges of the present invention specifically refer to each specific group covered. For example, the term "C1-4 alkyl" specifically refers to the independently disclosed methyl (i.e., C1 alkyl), ethyl (i.e., C2 alkyl), propyl (i.e., C3 alkyl), and butyl (i.e., C4 alkyl). For another example, an integer from 0 to 5 refers to the independently disclosed 0, 1, 2, 3, 4, and 5; an integer from 0 to 2 refers to the independently disclosed 0, 1, and 2.

[0072] In the present invention:

[0073] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the subject matter herein belongs.

[0074] It should be understood that one of ordinary skill in the art can select substituents and substitution patterns on the compounds of the present invention to produce chemically stable compounds. If substituted by more than one substituent group, it should be understood that these multiple groups can be on the same atom or on different atoms, as long as a stable structure is produced.

[0075] The term "substituted" or "substituted" means that a hydrogen atom on a group is replaced by a non-hydrogen atom group, but the valence requirements need to be met and the substitution generates a chemically stable compound, that is, a compound that does not spontaneously undergo transformations such as cyclization and elimination.

[0076] The terms "comprise," "include," and "contain" are used interchangeably to encompass not only closed-ended technical solutions but also semi-closed or open-ended technical solutions. In other words, the terms encompass "consisting of" or "consisting essentially of" technical solutions.

[0077] "Halogen" refers to F, Cl, Br or I.

[0078] Oxazole: Equivalent to oxazole, in English it is oxazole.

[0079] As used herein, the term "alkyl" refers to a straight-chain (i.e., unbranched) or branched saturated hydrocarbon group containing only carbon atoms and hydrogen atoms, or a combination of straight-chain and branched hydrocarbon groups, when used alone or as part of another substituent. When the alkyl group is preceded by a carbon number limit such as C1-8 alkyl, it means that the alkyl group contains 1-8 (e.g., 1, 2, 3, 4, 5, 6, 7, or 8) carbon atoms. For example, C1-4 alkyl refers to an alkyl group containing 1-4 carbon atoms, representative examples of which include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, or the like. The alkane may further be substituted with common groups such as OH, halogen, NO2, etc.

[0080] As used herein, the term "alkenyl," when used alone or as part of another substituent, refers to a linear or branched carbon chain radical having at least one carbon-carbon double bond. When the number of carbon atoms in an alkenyl group is specified, such as C2-8, it means that the alkenyl group contains 2 to 8 carbon atoms. For example, C2-4 alkenyl refers to an alkenyl group containing 2 to 4 carbon atoms, including ethenyl, propenyl, butenyl, butadienyl, and similar groups.

[0081] As used herein, the term "alkynyl," alone or as part of another substituent, refers to an aliphatic hydrocarbon group having at least one carbon-carbon triple bond. The alkynyl group may be straight-chain or branched, or a combination thereof. When the alkynyl group is preceded by a carbon atom number specification, such as C2-8 alkynyl, it means that the alkynyl group contains 2-8 carbon atoms. For example, the term "C2-4 alkynyl" refers to a straight-chain or branched alkynyl group having 2-4 carbon atoms, including ethynyl, propynyl, isopropynyl, butynyl, isobutynyl, sec-butynyl, tert-butynyl, or the like.

[0082] As used herein, the term "alkoxy" or "alkyloxy" refers to a RO- group, where R is an alkyl group, and alkyl is as defined above, and the alkoxy group is preceded by a carbon number, such as C1-8alkoxy, where the alkyl group has 1 to 8 carbon atoms. Representative examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, tert-butoxy, or the like.

[0083] As used herein, the term "cycloalkyl," when used alone or as part of another substituent, refers to a saturated or partially saturated monocyclic, bicyclic, or polycyclic (fused, bridged, or spirocyclic) ring system. When a cycloalkyl group is preceded by a carbon atom number specification such as C3-C10, it means that the cycloalkyl group has 3-10 carbon atoms. In some preferred embodiments, the term "C3-7 cycloalkyl" refers to a saturated or partially saturated monocyclic or bicyclic alkyl group having 3-7 carbon atoms, including cyclopropyl, cyclobutyl, cyclopentyl, cycloheptyl, or similar groups. "Spirocycloalkyl" refers to a bicyclic or polycyclic group in which the rings share a single carbon atom (called a spiro atom) between the rings. These rings may contain one or more double bonds, but none of the rings have a completely conjugated π electron system. "Fused cycloalkyl" refers to an all-carbon bicyclic or polycyclic group in which each ring in the system shares a pair of adjacent carbon atoms with the other rings in the system. One or more of the rings may contain one or more double bonds, but none of the rings have a completely conjugated π electron system. "Bridged cycloalkyl" refers to an all-carbon polycyclic group in which any two rings share two carbon atoms that are not directly connected. These may contain one or more double bonds, but neither ring has a completely conjugated pi electron system.

[0084] As used herein, the term "heterocycloalkyl," also known as heterocyclyl, when used alone or as part of another substituent, refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent in which one or more ring atoms are selected from nitrogen, oxygen, or sulfur. Non-limiting examples of monocyclic heterocyclyls include pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, and homopiperazinyl. Polycyclic heterocyclyls refer to heterocyclyls including spiro, fused, and bridged rings. "Spirocyclic heterocyclyl" refers to a polycyclic heterocyclic group in which each ring in the system shares an atom (called a spiro atom) with the other rings in the system, in which one or more ring atoms are selected from nitrogen, oxygen, or sulfur. "Fused-ring heterocyclyl" refers to a polycyclic heterocyclic group in which each ring in the system shares an adjacent pair of atoms with the other rings in the system, one or more rings may contain one or more double bonds, but none of the rings have a completely conjugated pi-electron system, and in which one or more ring atoms are selected from nitrogen, oxygen, or sulfur. "Bridged heterocyclyl" refers to a polycyclic heterocyclic group in which any two rings share two atoms that are not directly connected. These may contain one or more double bonds, but none of the rings has a completely conjugated π electron system, and one or more of the ring atoms are selected from nitrogen, oxygen or sulfur, and the remaining ring atoms are carbon. If a heterocyclyl contains both a saturated ring and an aromatic ring (for example, a saturated ring and an aromatic ring are fused together), the point of attachment to the parent must be on the saturated ring. Note: When the point of attachment to the parent is on the aromatic ring, it is called a heteroaryl, not a heterocyclyl. It should be understood in this article that a 3-10 membered heterocycloalkyl group refers to a heterocycloalkyl group having 3-10 atoms (such as 3, 4, 5, 6, 7, 8, 9 or 10). Examples of “3-7 membered heterocycloalkyl groups containing 1-3 ring heteroatoms independently selected from N, O, and S” include, but are not limited to, oxirane, thiol, aziridine, oxetane, N-heterocyclobutane, α-lactam ring, β-lactam ring, β-lactone, tetrahydrofuran, thiolane, pyrrolidine, pyrroline, dioxolane, oxazolidine, oxazoline, isoxazolidine, thiazolidine, isothiazolidine, thiazoline, imidazolidine, imidazoline, pyrazolidine, pyrazoline, tetrahydropyran, dihydropyran, pyran, piperidine, 1,4-dioxane, morpholine, piperazine, 1,4-oxazepane, 1,4-thiazepane, azepane, oxirane, thiepane, and the like.

[0085] As used herein, the term "aryl" refers to an all-carbon monocyclic or fused polycyclic (i.e., rings that share adjacent pairs of carbon atoms) group having a conjugated π electron system, when alone or as part of another substituent. When the term "aryl" is preceded by a carbon atom number specification, such as C6-C10 aryl, it means that the aryl group has 6-10 carbon atoms, for example, phenyl and naphthyl. The aryl ring may be fused to other cyclic groups (including saturated or unsaturated rings), but cannot contain heteroatoms such as nitrogen, oxygen, or sulfur. The point of attachment to the parent group must be on a carbon atom on the ring having a conjugated π electron system.

[0086] As used herein, the term "heteroaryl" refers to an aromatic heterocyclic ring system having one, two or more (preferably 1, 2, 3 or 4) heteroatoms, either alone or as part of another substituent, which may be a single ring (monocyclic) or a polycyclic ring (bicyclic, tricyclic or polycyclic) fused together or covalently linked, wherein the heteroatoms referred to herein include oxygen, sulfur and nitrogen. When the heteroaryl group is limited by the number of ring atoms, it refers to the number of ring atoms in the heteroaryl group, including carbon atoms and heteroatoms, for example, a 5-11 membered aryl group refers to a ring atom number of 5-11. The heteroaryl ring may be fused to an aryl, heterocycloalkyl or cycloalkyl ring, wherein the ring connected to the parent structure is the heteroaryl ring. Examples of “5-11 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O and S” include, but are not limited to, furan, thiophene, pyrrole, oxazole, isoxazole, thiazole, isothiazole, pyrazole, imidazole, 1,2,3-triazole, 1,2,4-triazole, oxadiazole, thiadiazole, pyridine, pyridine, pyrimidine, pyrimidine, pyridazine, pyridazine, pyrazine, pyrazine, triazine, benzazetidine, benzoβ-lactam ring, benzoβ-lactone, benzoxazoline, benzofuran, benzothiophene, indole, indazole, isoindazole, benzimidazole, benzothiazole, benzisothiazole, benzoxazole, benzisoxazole, benzotriazole, benzoxadiazole, benzothiadiazole, benzodioxane, benzothiadiazole, benzodioxane, ring, benzomorpholine, benzopiperidine, benzopyran, benzopyridine, benzopyrimidine, benzotriazine, benzazepine, benzazepine, pyridoazetidine, pyridoβ-lactam ring, pyridoβ-lactone, pyridoxadiazole, pyridofuran, pyridothiophene, pyridopyrrole, pyridopyrazole, pyridoimidazole, pyridothiazole, pyridoisothiazole, pyridoxazole, pyridoisoxazole, pyridotriazole, pyridoxadiazole, pyridothiadiazole, pyridodioxane, pyridomorpholine, pyridopiperidine, pyridopyran, pyridopyridine, pyridopyrimidine, pyridotriazine, pyridoazepine, furopyrrole, thiazolopyrimidine, and the like.

[0087] The compounds of the present invention may be asymmetric, for example, having one or more stereocenters. All stereoisomers, such as enantiomers and diastereomers, are included within the scope of the present invention unless otherwise indicated. In the present invention, compounds containing asymmetrically substituted carbon atoms can be isolated in either optically active or racemic forms. Various methods for preparing optically active forms are known in the art, for example, by resolving a racemic mixture or by stereoselective synthesis.

[0088] The present invention also includes hydrates and solvates of the compounds.

[0089] The present invention also includes all forms of the compounds wherein the atoms are isotopes. Isotopes include all atoms having the same atomic number but different mass numbers. For example, isotopes of hydrogen include deuterium.

[0090] The present invention also includes prodrugs of the compounds. "Prodrugs" refer to compounds obtained by structural modification of the compounds that are inactive or less active outside the patient's body, but release the compound through enzymatic or non-enzymatic conversion in the patient's body to exert its pharmacological effect.

[0091] "Independent of each other" or "independently" means that the features described are not interrelated with other features.

[0092] The phrase "substituents at any position on the ring" indicates that the substituents are located at any substitutable position on the ring. The substituents are located at the ortho, meta, or para position of the ring relative to the main chain.

[0093] The term "pharmaceutically acceptable salt" includes those derived from suitable inorganic and organic acids. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid, or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid, or salts formed by other methods known in the art such as ion exchange. DETAILED DESCRIPTION

[0094] The following terms or abbreviations are used in the examples:

[0095] DIAD: diisopropyl azodicarboxylate

[0096] LDA: lithium diisopropylamide

[0097] LiHMDS: Lithium bis(trimethylsilyl)amide

[0098] Pd(dppf)2Cl2:[1,1-bis(diphenylphosphino)ferrocene]palladium(II) dichloride dichloromethane complex

[0099] X-phos: 2-dicyclohexylphosphino-2,4,6-triisopropylbiphenyl

[0100] D2(Pin)2-Pinacol Borate

[0101] The present invention is further described below with reference to the following examples. It should be noted that the examples are not intended to limit the scope of protection of the present invention, and those skilled in the art will understand that any improvements and variations based on the present invention are within the scope of protection of the present invention.

[0102] The conventional reagents used in the following examples are all commercially available, and the biological experiments performed are all conventional biological experiments in the art and can be performed according to the instructions in the corresponding experimental manual or kit instructions.

[0103] The structures of the compounds of the present invention are determined by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS). NMR chemical shifts (δ) are given in parts per million (ppm). NMR measurements were performed using a Bruker AVANCE NEO400 NMR spectrometer, using deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol (CD3OD), and deuterated chloroform (CDCl3) as the solvents, with tetramethylsilane (TMS) as the internal standard.

[0104] Liquid chromatography-mass spectrometry (LC-MS) was performed using an Agilent 1260 HPLC coupled with a 6125B MS mass spectrometer. The most commonly used LCMS elution conditions were: 0-95% (0.01% TFA) for 3 minutes. Method: Phase A: 0.01% TFA water: acetonitrile = 90:10; Phase B: 0.01% TFA acetonitrile; Gradient: 0% B from 0-0.2 minutes; 0% B from 0.2-1.5 minutes; 95% B from 1.5-2.4 minutes; 95% B from 2.4-2.9 minutes; 0% B from 2.9-3.5 minutes; Flow rate: 2 ml / min; Column temperature: 40°C; Column: Waters CORTECS C18+ 2.7 μm, 4.6×30 mm

[0105] Example 1

[0106] Preparation of compound N-(2,6-dichlorophenyl)-4-methoxy-2-(1-(1-methylpyrrolidin-3-yl)-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (1)

[0107]

[0108] Step 1: Preparation of intermediate 4-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxylic acid methyl ester (1b)

[0109] Under an ice-water bath, sodium hydride (0.82 g, 20.44 mmol, 60% purity) was added to a solution of methyl 4-chloro-1H-pyrrolo[2,3-b]pyridine-5-carboxylate (1a) (2.87 g, 13.63 mmol) in dimethylformamide (25 mL) under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for half an hour and then poured into ice water (30 mL), followed by extraction with ethyl acetate (30 mL x 2). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and separated by silica gel column chromatography (ethyl acetate:petroleum ether = 0% to 6%) to obtain intermediate 1b (3.5 g, 75.4%) as a white solid. MS m / z (ESI): 341.2 [M+H] + .

[0110] Step 2: Preparation of intermediate 4-chloro-2-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxylic acid methyl ester (1c)

[0111] To a solution of LDA (4.84 mL, 9.68 mmol) in tetrahydrofuran was added intermediate 1b (2.2 g, 6.45 mmol) at -78°C under a nitrogen atmosphere, with the addition rate controlled to maintain the internal temperature of the solution above -78°C. The reaction mixture was stirred at -78°C for 1.0 hour, followed by the addition of a solution of iodine (3.28 g, 12.91 mmol) in tetrahydrofuran (2 mL), maintaining the temperature above -70°C. The reaction mixture was stirred at -78°C for an additional 1.0 hour, then poured into an ice-cooled saturated ammonium chloride solution (20 mL). The mixture was extracted with ethyl acetate (20 mL x 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography (ethyl acetate:petroleum ether = 2% to 10%) to afford intermediate 1c (0.3 g, 10.0%) as a white solid. MS m / z (ESI): 467.0 [M+H] + .

[0112] Step 3: Preparation of intermediate 4-chloro-2-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxylic acid (1d)

[0113] To a solution of Intermediate 1c (300 mg, 0.64 mmol) in tetrahydrofuran (2 mL) and water (1 mL) was added lithium hydroxide (53.94 mg, 2.24 mmol). The reaction mixture was stirred at 60°C for 3 hours, then the pH of the solution was adjusted to 7 with hydrochloric acid (1.0 M). The mixture was concentrated under reduced pressure to afford crude Intermediate 1d (350 mg, 83%) as a light yellow solid, which was used directly in the next reaction. MS m / z (ESI): 453.0 [M+H] + .

[0114] Step 4: Preparation of intermediate 4-chloro-2-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxylic acid chloride (1e)

[0115] To a solution of Intermediate 1d (330 mg, 0.6 mmol) in dichloromethane (5 mL) was added dropwise thionyl chloride (2 mL) under an ice-water bath and nitrogen atmosphere. The reaction solution was stirred at room temperature for 1.0 hour. The mixture was concentrated under reduced pressure to afford crude Intermediate 1e (330 mg) as a pale pink solid, which was used in the next step without further purification.

[0116] Step 5: Preparation of intermediate 4-chloro-N-(2,6-dichlorophenyl)-2-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (1f)

[0117] Under an ice-water bath and nitrogen atmosphere, a solution of LiHMDS (2.24 mL, 2.24 mmol) was added dropwise to a solution of 2,6-dichloroaniline (272.31 mg, 1.68 mmol) in tetrahydrofuran (4 mL). The reaction solution was stirred under an ice-water bath for 2.0 hours, followed by the addition of a solution of intermediate 1e (330 mg, 0.6 mmol, 85.6% purity) in dichloromethane (4 mL). The reaction solution was stirred under an ice-water bath for an additional 2.0 hours, then poured into water (20 mL). The mixture was extracted with ethyl acetate (20 mL x 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography (ethyl acetate:petroleum ether = 10% to 18%) to afford intermediate 1f (230 mg, 65%) as a yellow solid. MS m / z (ESI): 596.0 & 598.0 [M+H] + .

[0118] Step 6: Preparation of intermediate N-(2,6-dichlorophenyl)-2-iodo-4-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (1 g)

[0119] Under a nitrogen atmosphere, sodium methoxide (38.02 mg, 0.70 mmol) was added to a solution of intermediate 1f (210 mg, 0.35 mmol) in methanol (3 mL). The reaction solution was stirred at 60°C for 18 hours. After filtration and concentration, the product was separated by silica gel column chromatography (ethyl acetate:petroleum ether = 10% to 20%) to obtain intermediate 1g (150 mg, 72%) as a yellow solid. MS m / z (ESI): 592.1 [M+H] + .

[0120] Step 7: Preparation of intermediate N-(2,6-dichlorophenyl)-4-methoxy-2-(1-(1-methylpyrrolidin-3-yl)-1H-pyrazol-4-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (1h)

[0121] To a mixture of Intermediate 1g (150 mg, 0.25 mmol), 1-(1-methylpyrrolidin-3-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (233.97 mg, 0.51 mmol, 60% purity), and potassium carbonate (104.99 mg, 0.76 mmol) in mixed 1,4-dioxane (3 mL) and water (0.5 mL) was added Pd(dppf)2Cl2 (18.53 mg, 0.03 mmol) under a nitrogen atmosphere. The reaction mixture was stirred at 80°C for 16 hours and then poured into water (10 mL). The mixture was extracted with ethyl acetate (10 mL). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography (methanol:dichloromethane = 2-4%) to obtain Intermediate 1h (120 mg, 77.0%) as a white solid. MS m / z (ESI): 615.3 & 617.3 [M+H] + .

[0122] Step 8: Preparation of compound N-(2,6-dichlorophenyl)-4-methoxy-2-(1-(1-methylpyrrolidin-3-yl)-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (1)

[0123] To a solution of intermediate 1h (120 mg, 0.19 mmol) in dichloromethane was added trifluoroacetic acid (2 mL) at room temperature. The reaction solution was stirred at room temperature for 2.5 hours. The product was concentrated under reduced pressure, and the residue was dissolved in methanol (1 mL), followed by the addition of ammonia methanol solution (1 mL, 7 M). The resulting reaction mixture was stirred at room temperature for 0.5 hours and then concentrated under reduced pressure. The crude product was purified by preparative high-pressure liquid chromatography (Phenomenex Gemini 150 mm × 25 mm × 10 μm column (mobile phase: acetonitrile and water, 0.025% formic acid added)) to afford compound 1 (40 mg, 42.3%) as a white solid. MS m / z (ESI): 485.2 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ12.25(s,1H),9.85(s,1H),8.49(s,1H),8.36(s,1H),8.25-8.10(m,1H),8.02(s,1H),7.58(d,J=8.2Hz,2H),7.38(t ,J=8.2Hz,1H),7.10(d,J=1.8Hz,1H),4.99-4.90(m,1H),4.42(s,3H), 2.88-2.79(m,3H),2.45-2.39(m,1H),2.34(s,3H),2.14-2.03(m,1H).

[0124] Example 2: Preparation of Compound N-(2,6-dichlorophenyl)-4-(((1s,3s)-3-methoxycyclobutyl)amino)-2-(1-(1-methylpyrrolidin-3-yl)-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (2)

[0125]

[0126] Step 1: Preparation of intermediate 4-chloro-N-(2,6-dichlorophenyl)-2-(1-(1-methylpyrrolidin-3-yl)-1H-pyrazol-4-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (2a)

[0127] To a mixture of intermediate 1f (411 mg, 0.69 mmol), 1-(1-methylpyrrolidin-3-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (381.80 mg, 1.38 mmol), and potassium carbonate (190.4 mg, 1.4 mmol) in 1,4-dioxane (10 mL) and water (2 mL) was added Pd(dppf)2Cl2 (111.66 mg, 0.14 mmol) under a nitrogen atmosphere. The reaction mixture was stirred at 100°C for 16 hours and then poured into water (20 mL). The mixture was extracted with ethyl acetate (20 mL x 3). The organic phase was washed with saturated brine and water, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography (methanol / dichloromethane = 0-10%) to obtain intermediate 2a (400 mg, 93.7%) as a yellow oil. MS (ESI,) m / z: 619.2 [M+H] + .

[0128] Step 2: Preparation of intermediate N-(2,6-dichlorophenyl)-4-(((1s,3s)-3-methoxycyclobutyl)amino)-2-(1-(1-methylpyrrolidin-3-yl)-1H-pyrazol-4-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (2b)

[0129] To a solution of intermediate 2a (91 mg, 0.15 mmol) and cis-3-methoxycyclobutylamine hydrochloride (121 mg, 0.88 mmol) in n-butanol (1 mL) was added diisopropylethylamine (132.78 mg, 1.03 mmol) dropwise. The reaction mixture was stirred at 110°C in a microwave reactor for 10 hours, then diluted with water (20 mL) and extracted with ethyl acetate (20 mL x 3). The organic phase was washed with saturated brine and water, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography (methanol / dichloromethane = 0-10%) to afford intermediate 2b (80 mg, 79.6%) as a yellow oil. MS (ESI) m / z: 684.4 [M+H] + .

[0130] Step 3: Preparation of compound N-(2,6-dichlorophenyl)-4-(((1s,3s)-3-methoxycyclobutyl)amino)-2-(1-(1-methylpyrrolidin-3-yl)-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (2)

[0131] To a solution of intermediate 2b (80 mg, 0.12 mmol) in dichloromethane (0.5 mL) was added trifluoroacetic acid (0.3 mL). The reaction solution was stirred at room temperature for 2 hours and then concentrated under reduced pressure. The residue was dissolved in methanol (0.5 mL), and then ammonia methanol solution (0.5 mL, 7 M) was added. The resulting reaction mixture was stirred at room temperature for 2 hours and then concentrated under reduced pressure. The crude product was purified by preparative high-pressure liquid chromatography (Waters-XBridge-C18-5um-30×150mm, (mobile phase: acetonitrile and water, 0.025% formic acid added)) to give compound 2 (17.8 mg, 24.4%) as a white solid. MS (ESI,) m / z: 554.4 [M+H] + . 1H NMR(400MHz,DMSO-d6)δ11.90(s,1H),10.00(s,1H),9.10(s,1H),8.58(s,1H),8.51(s,1H),8.29(s, 1H),7.97(s,1H),7.58(d,J=8.1Hz,2H),7.42–7.36(m,1H),7.19(s,1H),6.81(d,J=2.1Hz,1H),6.65 (s,1H),5.33–5.32(m,1H),4.26–4.17(m,1H),3.81–3.71(m,1H),3.15(s,3H),2.87(s,1H),2.79–2. 76(m,1H),2.32(s,3H),2.02–1.99(m,4H),1.77–1.73(m,1H),1.46–1.44(m,1H),0.86–0.85(m,2H).

[0132] Example 3:

[0133] Preparation of compound 4-cyclobutoxy-N-(2,6-dichlorophenyl)-2-(1-(1-methylpyrrolidin-3-yl)-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (3)

[0134]

[0135] Step 1: Preparation of intermediate 4-cyclobutoxy-N-(2,6-dichlorophenyl)-2-(1-(1-methylpyrrolidin-3-yl)-1H-pyrazol-4-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (3a)

[0136] To a solution of cyclobutanol (302 mg, 4.19 mmol) in anhydrous dimethylformamide (5 mL) was added sodium hydride (168 mg, 4.19 mmol, 60% purity) under an ice-water bath. The reaction mixture was stirred at 0°C for 10 minutes, followed by the addition of intermediate 2a (130 mg, 0.21 mmol). The resulting mixture was stirred at 80°C for 18 hours, then poured into water (20 mL). The mixture was extracted with ethyl acetate (50 mL x 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by reverse phase column chromatography (acetonitrile / water: 10%-80%) to afford intermediate 3a (45 mg, 32.7%). MS (ESI) m / z: 655.4 & 657.4.

[0137] Step 2: Preparation of compound 4-cyclobutoxy-N-(2,6-dichlorophenyl)-2-(1-(1-methylpyrrolidin-3-yl)-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (3)

[0138] To a solution of intermediate 3a (45 mg, 0.07 mmol) in dichloromethane (1.0 mL) was added trifluoroacetic acid (1.5 mL). The reaction solution was stirred at room temperature for 1 hour and then concentrated under reduced pressure. The residue was dissolved in methanol (0.5 mL), and then ammonia methanol solution (2 mL, 7 M) was added. The resulting reaction mixture was stirred at room temperature for 2 hours and then concentrated under reduced pressure. The crude product was purified by preparative high-pressure liquid chromatography (Waters-XBridge-C18-5um-30×150mm, (mobile phase: A: 10 mM ammonium bicarbonate / water B: acetonitrile; A / B ratio: 66% to 95%) to afford compound 3 (25 mg, 0.05 mmol, 9.0%) as a light yellow solid. MS (ESI) m / z: 525.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δ11.77(br,1H),9.59(s,1H),9.04(s,1H),8.01(s,1H),7.88(s,1H),7.45(d,J=8.1Hz,2H),7.23(t,J=8.1Hz,1H),6.6 6(s,1H),5.68–5.47(m,1H),5.10(s,1H),3.25(s,2H),2.64(s,6H),2 .53–2.42(m,2H),2.36(s,1H),2.02–2.01(m,1H),1.90–1.75(m,3H).

[0139] Example 4: Preparation of compound N-(2,6-dichlorophenyl)-2-(1-(1-methylpyrrolidin-3-yl)-1H-pyrazol-4-yl)-4-(pyridin-3-yloxy)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (4)

[0140]

[0141] Step 1: Preparation of intermediate N-(2,6-dichlorophenyl)-2-iodo-4-(pyridin-3-yloxy)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (4a)

[0142] To a solution of intermediate 1f (200 mg, 0.34 mmol) and 3-hydroxypyridine (48 mg, 0.50 mmol) in anhydrous dimethylformamide (5 mL) was added potassium carbonate (93 mg, 0.67 mmol) at room temperature. The reaction mixture was stirred at 60°C for 18 hours and then diluted with ethyl acetate (60 mL) and petroleum ether (20 mL). The organic phase was washed with saturated brine and water, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography (ethyl acetate: petroleum ether: 0-45%) to afford intermediate 4a (100 mg, 45.5%) as a yellow solid. MS (ESI,) m / z: 655.2 [M+H] + .

[0143] Step 2: Preparation of intermediate N-(2,6-dichlorophenyl)-2-(1-(1-methylpyrrolidin-3-yl)-1H-pyrazol-4-yl)-4-(pyridin-3-yloxy)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (4b)

[0144] To a mixture of Intermediate 4a (100 mg, 0.15 mmol), 1-(1-methylpyrrolidin-3-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (127 mg, 0.46 mmol), and potassium carbonate (63 mg, 0.46 mmol) in a mixture of 1,4-dioxane (3 mL) and water (0.6 mL) was added Pd(dppf)2Cl2 (22 mg, 0.03 mmol) under a nitrogen atmosphere. The reaction mixture was stirred at 80°C for 2 hours. The mixture was concentrated under reduced pressure, and the residue was separated by silica gel column chromatography (methanol:dichloromethane = 0-20%) to afford Intermediate 4b (85 mg, 82.0%) as a yellow oil. MS (ESI) m / z: 680.4 [M+H] + .

[0145] Step 3: Preparation of compound N-(2,6-dichlorophenyl)-2-(1-(1-methylpyrrolidin-3-yl)-1H-pyrazol-4-yl)-4-(pyridin-3-yloxy)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (4)

[0146] To a solution of intermediate 4b (80 mg, 0.12 mmol) in dichloromethane (1.0 mL) was added trifluoroacetic acid (1.5 mL). The reaction solution was stirred at room temperature for 1 hour and then concentrated under reduced pressure. The residue was dissolved in methanol (0.5 mL), and then ammonia methanol solution (2 mL, 7 M) was added. The resulting reaction mixture was stirred at room temperature for 2 hours and then concentrated under reduced pressure. The crude product was purified by preparative high-pressure liquid chromatography (Waters-XBridge-C18-5um-30×150mm, (mobile phase: A: 10 mM ammonium bicarbonate / water B: acetonitrile; A / B ratio: 74% to 95%) to give compound 4 (10 mg, 15.4%) as a light yellow solid. MS (ESI) m / z: 547.9 & 549.9 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ12.45(s,1H),10.17(s,1H),8.55(s,1H),8.43(d,J=2.5Hz,1H),8.35(dd,J=4.1,1.9Hz,1H),8.32(s,1H),7.94(s,1H), 7.54(d,J=8.1Hz,2H),7.42–7.33(m,3H),6.12(s,1H),4.94–4.85(m,1H ),2.86–2.69(m,4H),2.40–2.35(m,1H),2.29(s,3H),2.09–2.01(m,1H).

[0147] Example 5: Preparation of compound N-(2,6-dichlorophenyl)-2-(1-(1-methylpyrrolidin-3-yl)-1H-pyrazol-4-yl)-4-(oxazol-2-ylmethoxy)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (5)

[0148]

[0149] Step 1: Preparation of intermediate N-(2,6-dichlorophenyl)-2-iodo-4-(oxazol-2-ylmethoxy)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (5a)

[0150] Under an ice-water bath, sodium hydride (60.3 mg, 1.51 mmol, 60% purity) was added to a solution of 1,3-oxazole-2-methanol (149 mg, 1.51 mmol) in anhydrous dimethylformamide. The reaction mixture was stirred at 0°C for 0.5 hours, and then intermediate 1f (300 mg, 0.50 mmol) was added. The resulting mixture was stirred at 60°C for 18 hours and then poured into water (30 mL). The mixture was extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography (methanol / dichloromethane: 0-20%) to obtain intermediate 5a (60 mg, 18.0%) as a yellow oil. MS m / z (ESI): 659.2 [M+H] + .

[0151] Step 2: Preparation of intermediate N-(2,6-dichlorophenyl)-2-(1-(1-methylpyrrolidin-3-yl)-1H-pyrazol-4-yl)-4-(oxazol-2-ylmethoxy)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (5b)

[0152] Using intermediate 5a as the starting material, the preparation and purification of intermediate 5b were similar to those of intermediate 4b. MS m / z (ESI): 682.4 [M+H] + .

[0153] Step 3: Preparation of compound N-(2,6-dichlorophenyl)-2-(1-(1-methylpyrrolidin-3-yl)-1H-pyrazol-4-yl)-4-(oxazol-2-ylmethoxy)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (5)

[0154] Starting from intermediate 5b, the preparation and purification of compound 5 were carried out according to the preparation of compound 4 (step 3). MS m / z (ESI): 552.2 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ12.32(s,1H),10.11(s,1H),8.47(s,1H),8.36(s,1H),8. 19(d,J=0.8Hz,1H),8.02(s,1H),7.56(d,J=8.0Hz,2H),7.37(t,J=8.0Hz,1H),7. 25(s,1H),7.10(d,J=2.0Hz,1H),5.89(s,2H),4.97(q,J=6.0,5.2Hz,1H),2.94–2 .80(m,3H),2.57-2.51(m,1H),2.48-2.38(m,1H),2.35(s,3H),2.15-2.05(m,1H).

[0155] Example 6: Preparation of compound N-(2,6-dichlorophenyl)-4-methoxy-2-(1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (6)

[0156]

[0157] Step 1: Preparation of intermediate 4-(4-bromo-1H-pyrazol-1-yl)-1-methylpiperidine (6a)

[0158] To a solution of N-methyl-4-hydroxypiperidine (1.0 g, 8.68 mmol) and 4-bromopyrrole (1.28 g, 8.68 mmol) in tetrahydrofuran (15 mL) were added triphenylphosphine (3.42 g, 13.02 mmol) and DIAD (2.58 mL, 2.63 g, 13.02 mmol). The reaction mixture was deoxygenated with nitrogen and then stirred at room temperature for 18 hours. The resulting mixture was poured into water (50 mL) and extracted with ethyl acetate (50 mL x 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography (methanol / dichloromethane: 0-10%) to afford Intermediate 6a (400 mg, 18.9%) as a yellow oil.

[0159] Step 2: Preparation of intermediate 1-methyl-4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)piperidine (6b)

[0160] To a solution of intermediate 6a (350 mg, 1.43 mmol) in 1,4-dioxane (5 mL) was added 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolan (0.56 mL, 2.15 mmol), Pd(dppf)2Cl2 (104.90 mg, 0.14 mmol), and potassium acetate (351.73 mg, 3.58 mmol). The reaction mixture was deoxygenated with nitrogen and then stirred at 90°C for 18 hours. The mixture was filtered, and the filter cake was washed with ethyl acetate (20 mL x 3). The combined organic phases were concentrated under reduced pressure, and the crude product was used directly in the next reaction without further purification.

[0161] Step 3: Preparation of intermediate N-(2,6-dichlorophenyl)-4-methoxy-2-(1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (6c)

[0162] To a mixture of Intermediate 1g (210 mg, 0.35 mmol), Intermediate 6b (490 mg, 1.68 mmol), and potassium carbonate (147 mg, 1.06 mmol) in mixed 1,4-dioxane (2 mL) and water (0.4 mL) was added Pd(dppf)2Cl2 (25.9 mg, 0.04 mmol) under a nitrogen atmosphere. The reaction mixture was deoxygenated with nitrogen and then stirred at 80°C for 2 hours. The mixture was poured into water (20 mL) and extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography (methanol / dichloromethane: 0-20%) to afford Intermediate 6c as a brown oil. MS m / z (ESI): 629.4 [M+H] + .

[0163] Step 4: Preparation of compound N-(2,6-dichlorophenyl)-4-methoxy-2-(1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (6)

[0164] Starting from intermediate 6c, the preparation and purification of compound 6 were carried out according to the preparation of compound 4 (step 3). MS m / z (ESI): 499.3 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ12.23(s,1H),9.85(s,1H),8.49(s,1H),8.32(s,1H),8.02(s,1H),7.58(d,J=8.1Hz,2H),7.38(t,J=8.1 Hz,1H),7.08(s,1H),4.41(s,3H),4.21–4.12(m,1H),2.87(d,J=11.1Hz,2H),2.22(s,3H),2.12–2.04(m,4H),2.00–1.94(m,2H).

[0165] Example 7: Preparation of compound N-(2,6-dichlorophenyl)-4-methoxy-2-(1-(1-methylpiperidin-3-yl)-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (7)

[0166]

[0167] Using intermediate 7a as the starting material, the preparation and purification process of compound 7 was referred to the preparation process of compound 6. MSm / z(ESI):500.0[M+H] + . 1 H NMR(400MHz,DMSO-d6)δ12.23(s,1H),9.84(s,1H),8.49(s,1H),8.37(s,1H) ,8.02(s,1H),7.58(d,J=8.0Hz,2H),7.38(t,J=8.0Hz,1H),7.07(s,1H),4.4 2(s,3H),4.36–4.28(m,1H),3.01(d,J=10.0Hz,1H),2.68(d,J=11.2Hz,1H), 2.32–2.20(m,4H),2.10–1.96(m,2H),1.81–1.71(m,2H),1.69–1.58(m,1H).

[0168] Example 8: Preparation of compound N-(2,6-dichlorophenyl)-2-(1-((1s,3s)-3-(dimethylamino)cyclobutyl)-1H-pyrazol-4-yl)-4-methoxy-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (8)

[0169]

[0170] Step 1: Preparation of intermediate tert-butyl (3-(4-bromo-1H-pyrazol-1-yl)cyclobutyl)carbamate (8a)

[0171] To a solution of tert-butyl (3-hydroxycyclobutyl)carbamate (1.00 g, 5.34 mmol) and 4-bromopyrazole (784 mg, 5.34 mmol) in tetrahydrofuran (10 mL) were added triphenylphosphine (2.11 g, 8.01 mmol) and DIAD (1.62 g, 8.01 mmol). The reaction mixture was stirred at room temperature for 18 hours. The mixture was concentrated under reduced pressure and separated by silica gel column chromatography (ethyl acetate / petroleum ether: 0-30%) to afford Intermediate 8a (950 mg, 56.3%) as a white solid. MS m / z (ESI): 316.0 & 318.0 [M+H] + .

[0172] Step 2: Preparation of intermediate (3-(4-bromo-1H-pyrazol-1-yl)cyclobutyl-1-amino) (8b)

[0173] To a solution of Intermediate 8a (950 mg, 3.00 mmol) in dichloromethane (10 mL) was added trifluoroacetic acid (2 mL). The reaction mixture was stirred at room temperature for 2 hours. It was concentrated under reduced pressure to afford crude Intermediate 8b (1.00 g, 50% purity, 77.2%) as a yellow oil, which was used directly in the next step without further purification. MS m / z (ESI): 216.0 & 218.0 [M+H] + .

[0174] Step 3: Preparation of intermediate cis-(3-(4-bromo-1H-pyrazol-1-yl)-N,N-dimethylcyclobutyl-1-amino) (8c)

[0175] Sodium cyanoborohydride (1.74 g, 27.77 mmol) was added to a solution of Intermediate 8b (1.20 g, 5.55 mmol) and formaldehyde (4.63 g, 55.53 mmol, 36% purity, 4.25 mL) in methanol (10 mL). The reaction mixture was stirred at room temperature for 18 hours and then concentrated under reduced pressure. The residue was separated by silica gel column chromatography (methanol / dichloromethane: 0-10%) to afford Intermediate 8c (1.03 g, 73.8%) as a light yellow oil. MS m / z (ESI): 244.0 & 246.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.10 (d, J = 0.8 Hz, 1H), 7.62 (d, J = 0.8 Hz, 1H), 2.85-2.75 (m, 2H), 2.74 (s, 6H), 2.70-2.61 (m, 2H).

[0176] Step 4: Preparation of the intermediate cis-N,N-dimethyl-3-(4-(4,4,5,5)-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)-cyclobutyl-1-amino (8d)

[0177] To a solution of Intermediate 8c (500 mg, 2.05 mmol) and 4,4,5,5-tetramethyl-2-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolan (1.04 g, 4.10 mmol) in 1,4-dioxane (10 mL) were added Pd2(dba)3 (375 mg, 0.41 mmol), X-phos (293 mg, 0.61 mmol), and potassium acetate (603 mg, 6.14 mmol). The reaction mixture was stirred at 85°C for 2 hours. The mixture was filtered, and the filtrate was concentrated under reduced pressure to give crude Intermediate 8d (500 mg, 1.72 mmol, 50% purity, 83.8%), which was used in the next step without further purification. MS m / z (ESI): 292.2 [M+H] + .

[0178] Step 5: Preparation of intermediate N-(2,6-dichlorophenyl)-2-(1-(cis-3-(dimethylamino)cyclobutyl)-1H-pyrazol-4-yl)-4-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (8e)

[0179] To a mixture of Intermediate 8d (197 mg, 0.68 mmol), Intermediate 1g (200 mg, 0.34 mmol), and potassium carbonate (140 mg, 1.01 mmol) in 1,4-dioxane (6 mL) and water (1.0 mL) was added Pd(dppf)2Cl2 (24.71 mg, 0.03 mmol) under a nitrogen atmosphere. The reaction mixture was stirred at 80°C for 2 hours. The mixture was concentrated under reduced pressure, and the residue was separated by silica gel column chromatography (methanol:dichloromethane = 0-20%) to afford Intermediate 8e (100 mg, 47.0%) as a yellow oil. MS m / z (ESI): 629.4 [M+H] + .

[0180] Step 6: Preparation of compound N-(2,6-dichlorophenyl)-2-(1-((1s,3s)-3-(dimethylamino)cyclobutyl)-1H-pyrazol-4-yl)-4-methoxy-1H-pyrrolo[2,3-b]pyridine-5-carboxamide (8)

[0181] To a solution of intermediate 8e (80.0 mg, 0.13 mmol) in dichloromethane (3.0 mL) was added trifluoroacetic acid (1.5 mL). The reaction solution was stirred at room temperature for 12 hours and then concentrated under reduced pressure. The residue was dissolved in methanol (1.5 mL), and ammonia methanol solution (1.5 mL, 7 M) was added. The resulting reaction mixture was stirred at room temperature for 12 hours and then concentrated under reduced pressure. The crude product was purified by preparative high-pressure liquid chromatography (Phenomenex Gemini 150 mm × 25 mm × 10 μm column, mobile phase: acetonitrile and water mixed with 0.1% ammonium bicarbonate) to afford compound 8 (20.5 mg, 32.1%) as a white solid. MS m / z (ESI): 499.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ12.23(s,1H),9.84(s,1H),8.50(s,1H),8.369(s,1H),8.05(s,1H),7.59(d,J=8.0Hz,2H),7. 39(t,J=8.0Hz,1H),7.12(s,1H),4.66-4.54(m,1H),4.43(s,3H),2.74-2.53(m,3H),2.37–2.26(m,2H),2.12(s,6H).

[0182] Examples 9-20

[0183] The compounds of Examples 9-20 were prepared using a method similar to that of Examples 1-8 with different starting materials. The structures and confirmation parameters of the 12 compounds are shown in the table below.

[0184]

[0185]

[0186]

[0187]

[0188] Biological activity test

[0189] 1. pCDK1 Activity Assay

[0190] pCDK1 is a direct substrate of WEE1. By detecting the activity state and phosphorylation level of pCDK1 Y15, the inhibitory level of the compound on WEE1 can be determined.

[0191] Experimental Methods: Widr cells were cultured in a T75 flask with EMEM + 10% FBS + 1% P / S. After two days, the medium was removed and rinsed once with PBS. The cells were then incubated with 3 mL of TrypLE™ Express Enzyme at room temperature or 37°C for 2-3 minutes until the cells detached. 9 mL of fresh medium was added, the cells were suspended, and centrifuged at 1000 rpm for 5 minutes at room temperature. The supernatant was discarded, the cells were resuspended in 3 mL of fresh medium, and the cells were returned to the T75 flask for further culture or analyzed in a culture dish for intracellular western blot analysis. Suitable cells were plated in a 384-well flat-bottom TC-treated microplate and incubated overnight at 37°C with 5% CO2. 60 nL of cisplatin was added to the cells using an Echo655. After 24 hours of incubation, 40 nL of serial dilutions were added and the cells were incubated at 37°C with 5% CO2 for 6 hours. After fixing the cells with 40 μL of 8% fixative, wash twice with 40 μL of PBS. Add 40 μL of methanol to each well and incubate at room temperature for 10 minutes. Then, wash twice with 40 μL of PBS and incubate with 20 μL of Li-Cor blocking buffer at room temperature for 1 hour. Remove the blocking buffer and add the primary antibody cocktail (4 μL of Phospho-cdc2 (Tyr15) antibody and 2 μL of GAPDH (D4C6R) mouse mAb) and incubate overnight at 4°C. After washing three times with PBST (PBS containing 0.05% Tween-20), the secondary antibody cocktail was added (2 μL IRDye 680RD goat anti-mouse IgG (0.5 mg) and 2 μL IRDye 800CW goat anti-rabbit IgG (H+L) (0.5 mg) were added to 4 mL of blocking buffer, and incubated at room temperature in the dark for 1 hour). After washing three times with PBST, the plate was inverted and centrifuged at 1000 rpm for 1 minute, and the plate was scanned with Odyssey CLx.

[0192] 2. PLK1 Selectivity Assay

[0193] PLK1 primarily regulates kinases acting on the B1 and CDK1 proteins in cells during the G2 / M phase. During mitosis, cohesin is removed from the sister chromatid arms, activating the APC / C ubiquitin E3 ligase complex, thereby affecting spindle formation and cytoplasmic division during mitosis. Overexpression of PLK1 can cause a variety of cancers (breast, bladder, prostate, pancreatic, papillary thyroid, ovarian, and head and neck cancers, as well as non-small cell lung cancer and non-Hodgkin's lymphoma).

[0194] Experimental Method: Volasertib, a positive drug, was serially diluted 3-fold in DMSO in a 384-well plate, with a final starting concentration of 100 nM. Test compounds were serially diluted 3-fold in DMSO, with a final starting concentration of 30 μM. 50 nL of the diluted compound was transferred to a 384-well plate using an echophoresis system, ensuring a 1% DMSO concentration.

[0195] Transfer 2.5 μL of 2× PLK1 enzyme solution to a 384-well plate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 10 minutes. Transfer 2.5 μL of 2× ATP and PLKtide solution to a 384-well plate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 60 minutes. Transfer 4 μL of ADP-Glo ​​Reagent to a 384-well plate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 40 minutes. Transfer 8 μL of ADP-Glo ​​Detection reagent to a 384-well plate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 40 minutes. Read the RLU (Relative Luminescence Unit) signal using a BMG microplate reader. Signal intensity is used to indicate the activity of the PLK1 enzyme.

[0196] Example pCDK1 Ki(Wee1)(μM) PLK1 enzyme (μM) 1 0.56 >30 3 0.56 >30 5 0.53 29.7 6 0.35 >30 7 0.70 8 0.45 11 0.52 12 0.489 13 0.632 17 0.607 19 0.547

[0197] The above describes the embodiments of the present invention. However, the present invention is not limited to the above embodiments. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included in the scope of protection of the present invention.

Claims

1. A compound having the structure of Formula I, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, in, n is an integer from 0 to 5; R1 is independently selected from halogen, -OH, -CN, -NH2, -C1-8 alkyl, -C2-8 alkenyl, -C2-8 alkynyl, -C1-8 alkoxy, -C2-8 alkenyloxy, -C2-8 alkynyloxy, and the alkyl, alkenyl, alkynyl, alkoxy, alkenyloxy and alkynyloxy groups are optionally substituted with one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C1-4 alkoxy; R2 is selected from H, halogen, -OH, -CN, -NH2, -C1-8 alkyl, -C2-8 alkenyl, -C2-8 alkynyl, -OR7, -N(R7)R8, wherein the alkyl, alkenyl, and alkynyl groups are optionally substituted with one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, -C1-4 alkoxy, C3-10 cycloalkyl, 3-10 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, and S, C6-10 aryl, and 5-11 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S; R7 is selected from -C1-8 alkyl, -C2-8 alkenyl, -C2-8 alkynyl, -(CH2) m -C3-10 cycloalkyl, -(CH2) p -3-10 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, S, -(CH2) r -C6-10 aryl, -(CH2) q - a 5-11 membered heteroaryl group containing 1-3 ring heteroatoms independently selected from N, O, and S, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are optionally substituted with one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, and -C1-4 alkoxy; R8 is selected from H, -C1-8 alkyl, -C2-8 alkenyl, and -C2-8 alkynyl, wherein the alkyl, alkenyl, and alkynyl groups are optionally substituted with one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, and -C1-4 alkoxy; m, p, r, and q are independently integers of 0 to 4; R3, R4, R6 are independently selected from H, halogen, -OH, -CN, -NH2, -C1-8 alkyl, -C2-4 alkenyl, -C2-8 alkynyl, -C1-8 alkoxy; R5 is selected from C3-10 cycloalkyl, 3-10 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, and S, C6-10 aryl, and 5-11 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S, wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted by one or more independently selected from =O, halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, -OR9, -C(O)R9, -N(R9)R 10 wherein the alkyl, alkenyl and alkynyl groups are optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl and -C1-4 alkoxy; R9 is selected from -C1-8 alkyl, -C2-8 alkenyl and -C2-8 alkynyl, wherein the alkyl, alkenyl and alkynyl groups are optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl and -C1-4 alkoxy; R 10 Selected from H, -C1-8 alkyl, -C2-8 alkenyl, -C2-8 alkynyl, and the alkyl, alkenyl, and alkynyl are optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, and -C1-4 alkoxy. Preferably, in Formula I, n is an integer from 0 to 5; R1 is independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, -C1-4 alkoxy, -C2-4 alkenyloxy, -C2-4 alkynyloxy, and the alkyl, alkenyl, alkynyl, alkoxy, alkenyloxy and alkynyloxy groups are optionally substituted with one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C1-4 alkoxy; R2 is selected from H, halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, -OR7, -N(R7)R8, wherein the alkyl, alkenyl, and alkynyl groups are optionally substituted with one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, -C1-4 alkoxy, C3-7 cycloalkyl, 3-7 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, and S, C6-10 aryl, and 5-11 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S; R7 is selected from -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, -(CH2) m -C3-7 cycloalkyl, -(CH2) p -3-7 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, S, -(CH2) r -C6-10 aryl, -(CH2) q -5-11 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are optionally substituted with one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, and -C1-4 alkoxy; R8 is selected from H, -C1-4 alkyl, -C2-4 alkenyl, and -C2-4 alkynyl, wherein the alkyl, alkenyl, and alkynyl groups are optionally substituted with one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, and -C1-4 alkoxy; m, p, r, and q are independently integers of 0 to 4; R3, R4, R6 are independently selected from H, halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, -C1-4 alkoxy; R5 is selected from C3-7 cycloalkyl, 3-7 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, and S, C6-10 aryl, and 5-11 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S, wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted by one or more independently selected from =O, halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, -OR9, -C(O)R9, -N(R9)R 10 wherein the alkyl, alkenyl and alkynyl groups are optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl and -C1-4 alkoxy; R9 is selected from -C1-4 alkyl, -C2-4 alkenyl and -C2-4 alkynyl, wherein the alkyl, alkenyl and alkynyl groups are optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl and -C1-4 alkoxy; R 10 Selected from H, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, and the alkyl, alkenyl, and alkynyl are optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C2-4 alkenyl, -C2-4 alkynyl, and -C1-4 alkoxy.

2. The compound according to claim 1, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, wherein: The compound comprises one or more characteristics selected from the group consisting of: (1) n is an integer from 0 to 2; (2) R1 is independently selected from F, Cl, Br, -OH, -CN, -NH2, -C1-4 alkyl, -C1-4 alkoxy, and the alkyl and alkoxy groups are optionally substituted with one or more groups independently selected from F, Cl, Br, -OH, -CN, -NH2, -C1-4 alkyl, and -C1-4 alkoxy; (3) R2 is selected from halogen, -C1-4 alkyl, -OR7, -N(R7)R8, wherein the alkyl group is optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C1-4 alkoxy, C3-7 cycloalkyl, 3-7 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, and S, -C6-10 aryl, or 5-6 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S; R7 is selected from -C1-4 alkyl, -(CH2) m -C3-7 cycloalkyl, -(CH2) p -3-7 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, S, -(CH2) r -C6-10 aryl, -(CH2) q - a 5-6 membered heteroaryl group containing 1-3 ring heteroatoms independently selected from N, O, and S, wherein the alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are optionally substituted with one or more groups independently selected from -OH, -C1-4 alkyl, and -C1-4 alkoxy; R8 is selected from H, -C1-4 alkyl, and the alkyl group is optionally substituted with one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, and -C1-4 alkoxy; m, p, r, and q are independently integers of 0 to 2; (4) R3, R4, and R6 are independently selected from H, halogen, -C1-4 alkyl, and -C1-4 alkoxy; (5) R5 is selected from C3-7 cycloalkyl, 3-7 membered heterocycloalkyl containing 1 ring heteroatom of N or O, C6-10 aryl, 5-6 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, S, wherein the cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted by one or more independently selected from =O, -C1-4 alkyl, -OR9, -C(O)R9, -N(R9)R 10 wherein the alkyl group is optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C1-4 alkoxy; R9 is selected from -C1-4 alkyl, wherein the alkyl group is optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C1-4 alkoxy; R 10 Selected from -C1-4 alkyl, wherein the alkyl is optionally substituted by one or more groups independently selected from halogen, -OH, -CN, -NH2, -C1-4 alkyl, -C1-4 alkoxy.

3. The compound according to claim 1, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, wherein: The compound has the structure shown in formula I-1, in: Each R1 is independently selected from F, Cl, Br, CH3, CH2CH3, CH3O; R2 is selected from halogen, -C1-4 alkyl, -OR7, -N(R7)R8, wherein the alkyl group is optionally substituted by one or more groups independently selected from halogen, -C1-4 alkyl, -C1-4 alkoxy, C3-7 cycloalkyl, 3-7 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, and S, C6-10 aryl, and 5-6 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S; R7 is selected from -C1-4 alkyl, -(CH2) m -C3-7 cycloalkyl, -(CH2) p -3-7 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, S, -(CH2) r -C6-10 aryl, -(CH2) q - a 5-6 membered heteroaryl group containing 1-3 ring heteroatoms independently selected from N, O, and S, wherein the alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are optionally substituted with one or more groups independently selected from halogen, -OH, -C1-4 alkyl, and -C1-4 alkoxy; R8 is selected from H, -C1-4 alkyl; m, p, r, and q are independently integers of 0 to 1; R3, R4, R6 are independently selected from H, halogen, -C1-4 alkyl, -C1-4 alkoxy; R5 is selected from C3-7 cycloalkyl, 4-6 membered heterocycloalkyl containing 1 ring heteroatom selected from N or O, 5-6 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, S, and the cycloalkyl, heterocycloalkyl and heteroaryl are optionally substituted by one or more independently selected from =O, -C1-4 alkyl, -OR9, -C(O)R9, -N(R9)R 10 wherein the alkyl group is optionally substituted by one or more groups independently selected from halogen, -C1-4 alkyl, -C1-4 alkoxy; R9 is selected from -C1-4 alkyl; R 10 Selected from -C1-4 alkyl.

4. The compound according to claim 3, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, wherein: The compound of formula I-1 comprises one or more characteristics selected from the group consisting of: (i) R3, R4, and R6 are H; (ii) R2 is selected from F, Cl, Br, -C1-4 alkyl, -OR7, -N(R7)R8, wherein the alkyl group is optionally substituted by one or more groups independently selected from halogen, -C1-4 alkyl, -C1-4 alkoxy, C3-7 cycloalkyl, 3-7 membered heterocycloalkyl containing 1-3 ring heteroatoms independently selected from N, O, and S, C6-10 aryl, and 5-6 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S; R7 is selected from -C1-4 alkyl, -C3-7 cycloalkyl, -(CH2) q -5-6 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S, wherein the cycloalkyl group is optionally substituted by a group selected from -C1-4 alkoxy, and the alkyl group is optionally substituted by one or more groups independently selected from F, Cl, and Br; R8 is selected from H, -C1-4 alkyl; q is an integer from 0 to 1; (iii) R5 is selected from C3-7 cycloalkyl, 4-6 membered heterocycloalkyl containing one selected from N or O, 5-6 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, S, said cycloalkyl being substituted by one -N(R9)R 10 The heterocycloalkyl group is connected to the pyrazole group through a ring carbon atom, and the heterocycloalkyl group is optionally substituted by one or more groups independently selected from =O, -C1-4 alkyl, -OR9, and -C(O)R9, wherein the alkyl group is optionally substituted by one or more groups independently selected from F, Cl, Br, -C1-4 alkyl, and -C1-4 alkoxy; R9 is selected from -C1-4 alkyl; R 10 Selected from -C1-4 alkyl.

5. The compound according to claim 3, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, wherein: In formula I-1: R3, R4, and R6 are H; R2 is selected from F, Cl, Br, -C1-4 alkyl, -OR7, -N(R7)R8, wherein the alkyl group is optionally substituted by one or more groups independently selected from halogen, -C1-4 alkyl, -C1-4 alkoxy; R7 is selected from -C1-4 alkyl, -C3-7 cycloalkyl, -(CH2) q -5-6 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S, wherein the cycloalkyl group is optionally substituted by a group selected from -C1-4 alkoxy, and the alkyl group is optionally substituted by one or more groups independently selected from F, Cl, and Br; R8 is selected from H, -C1-4 alkyl; q is an integer from 0 to 1; R5 is selected from C4-6 cycloalkyl, containing one 4-6 membered heterocycloalkyl selected from N or O, said cycloalkyl being replaced by one -N(R9)R 10 wherein the heterocycloalkyl group is connected to the pyrazole group via a ring carbon atom, and the heterocycloalkyl group is optionally substituted by one or more groups independently selected from =O, -C1-4 alkyl, and -C(O)R9. When the heterocycloalkyl group is an N-containing heterocycloalkyl group, if it has a -C1-4 alkyl or -C(O)R9 substituent, the substituent is located on the ring N atom, wherein the alkyl group is optionally substituted by one or more groups independently selected from F, Cl, Br, and -C1-4 alkoxy; R9 is selected from -C1-4 alkyl; R 10 Selected from -C1-4 alkyl. Preferably, R3, R4, and R6 are H; R2 is selected from -OR7, -N(R7)R8; R7 is selected from -C1-4 alkyl, -C3-7 cycloalkyl, -(CH2) q -5-6 membered heteroaryl containing 1-3 ring heteroatoms independently selected from N, O, and S, wherein the alkyl group is optionally substituted by one or more groups independently selected from F, Cl, and Br, and the cycloalkyl group is optionally substituted by a group selected from -C1-4 alkoxy; R8 is selected from H, -C1-4 alkyl; q is an integer from 0 to 1; R5 is selected from C4-6 cycloalkyl (preferably cyclobutyl, cyclopentyl, cyclohexyl), 5-6 membered heterocycloalkyl containing 1 O ring heteroatom (preferably ), a 4-6 membered heterocycloalkyl containing one N ring heteroatom (preferably ), the cycloalkyl group is replaced by a -N(R9)R 10 The heterocycloalkyl group is connected to the pyrazole group through a ring carbon atom, the ring N heteroatom of the heterocycloalkyl group is optionally substituted by a -C1-4 alkyl group, and the alkyl group is optionally substituted by one or more groups independently selected from F, Cl, Br, and -C1-4 alkoxy; R9 is selected from -C1-4 alkyl; R 10 Selected from -C1-4 alkyl.

6. The compound according to claim 3, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, wherein: In formula I-1: R3, R4, and R6 are H; R2 is selected from -OR7, -N(R7)R8, R7 is selected from -C1-4 alkyl, -C3-7 cycloalkyl, -(CH2) q - a 5-6 membered heteroaryl group containing 1-3 ring heteroatoms independently selected from N, O, and S, wherein the cycloalkyl group is optionally substituted by a group selected from -C1-4 alkoxy; R8 is selected from H, -C1-4 alkyl; q is an integer from 0 to 1; R5 is selected from C4-6 cycloalkyl (preferably cyclobutyl, cyclopentyl, cyclohexyl), 4-6 membered heterocycloalkyl containing 1 N ring heteroatom (preferably ), the cycloalkyl group is replaced by a -N(R9)R 10 The heterocycloalkyl group is connected to the pyrazole group through a ring carbon atom, and the ring N heteroatom of the heterocycloalkyl group is optionally substituted by a -C1-4 alkyl group; R9 is selected from -C1-4 alkyl; R 10 Selected from -C1-4 alkyl.

7. The compound according to claim 1, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, wherein: The compound of formula I is selected from the following compounds:

8. A method for preparing the compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, comprising reacting a compound of formula A with a compound of formula B, and then removing the group SEM and / or further modifying the group as needed to form a compound of formula I, wherein R1-R6 and n are as defined in any one of claims 1 to 7, SEM is 2-(trimethylsilyl)ethoxy)methyl, and X is a leaving group, preferably a halogen, more preferably iodine; 9. An in vitro non-therapeutic and non-diagnostic method for inhibiting WEE1, comprising the steps of contacting WEE1 or a cell expressing WEE1 with a compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof.

10. Use of the compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof in the preparation of a medicament for treating diseases mediated by WEE1 overexpression or hyperactivity; preferably, the diseases mediated by WEE1 overexpression or hyperactivity are tumors, including but not limited to: breast cancer, ovarian cancer, liver cancer, cervical cancer, lung cancer, squamous cell carcinoma, colorectal cancer, gastric cancer, glioblastoma, diffuse intramural glioma, and melanoma.

11. A pharmaceutical composition, characterized in that Comprising the compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof; preferably, the pharmaceutical composition further contains a pharmaceutically acceptable carrier.

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