Small-molecule inhibitor targeting protein kinase, membrane‑associated tyrosine / threonine (pkmyt1) and use thereof

Abstract

Provided in the present invention are a PKMYT1 inhibitor compound as represented by structural formula (I), a prodrug, an optical isomer, a tautomer, an isotope derivative, a solvate, a pharmaceutically acceptable salt and a pharmaceutical composition thereof, and a preparation method therefor and the use thereof. In the present invention, the high efficiency, safety, and feasibility of the compound of the present invention in targeted treatment are demonstrated by means of multi-dimensional validation of enzyme activity inhibition, cytotoxicity, safety and in-vivo efficacy. Moreover, the compound of the present invention has a very strong PKMYT1 inhibitory effect, and thus can be used as a drug for treating diseases associated with PKMYT1 overexpression.

Description

Small molecule inhibitors targeting membrane-associated tyrosine / threonine protein kinase (PKMYT1) and applications thereof TECHNICAL FIELD

[0001] The present application belongs to the field of chemical drugs, and particularly relates to a small molecule inhibitor targeting membrane-associated tyrosine / threonine protein kinase (PKMYT1), a preparation method and applications thereof. BACKGROUND

[0002] Membrane-associated tyrosine / threonine protein kinase 1 (PKMYT1) is a member of the Wee protein kinase family. The Wee protein kinase family includes three members: Wee1, Wee2 and PKMYT1. Wee2 mainly regulates the development of germ cells, while Wee1 and PKMYT1 play important roles in cell mitosis regulation. Wee1 is mainly located in the nucleus, while PKMYT1 is located in the cytoplasm and is associated with the Golgi and endoplasmic reticulum through membrane anchoring.

[0003] During the DNA replication process of cells, mismatches and defects often occur. In order to maintain the stability of the genome, normal cells repair these errors through checkpoints in the cell cycle. Among them, the G1 / S checkpoint can monitor DNA damage and prevent damaged DNA from entering mitosis, thereby avoiding the transmission of errors to daughter cells. However, due to mutations in the P53 gene, many tumor cells have lost the function of the G1 / S checkpoint. In order to ensure the integrity of the genome, tumor cells rely on the G2 / M checkpoint to repair DNA damage. The key factors for cells to enter the mitotic phase include the activation of protein kinase Cdk1 (cyclin-dependent kinase 1), the accumulation of CycB, and the nuclear-cytoplasmic shuttling of the Cdk1 / CycB complex. In this process, PKMYT1 plays an important role. PKMYT1 can phosphorylate Thr14 and Tyr15 on Cdk1, thereby inhibiting the activity of Cdk1 and hindering its binding with CycB. In addition, PKMYT1 also binds to the Cdk1 / CycB complex and isolates it in the cytoplasm, preventing the complex from entering the nucleus and hindering the progress of nuclear-cytoplasmic shuttling. The absence of PKMYT1 will interfere with the normal function of the G2 / M checkpoint, causing CDK1 to be over-activated, and thus leading to apoptosis or irreversible damage, ultimately resulting in the death of tumor cells. Therefore, as a key regulatory factor of the G2 / M checkpoint, PKMYT1 is considered as a potential target for tumor treatment.

[0004] Increasingly, studies have shown that PKMYT1 is closely related to the occurrence and development of tumors. For example, in gastric cancer, non-small cell lung cancer, hepatocellular carcinoma, glioblastoma, neuroblastoma and colorectal cancer and other tumors, overexpression of PKMYT1 is associated with increased tumor invasiveness, metastatic ability and chemotherapy resistance. In addition, recent studies have found that CCNE1 amplification and FBXW7 mutations have special sensitivity to the loss of PKMYT1. In ovarian cancer, about 20% of tumors detect CCNE1 amplification, which is largely mutually exclusive with homologous recombination deficiency, and is more common in platinum drug-resistant tumors. However, due to the limited treatment options for tumors caused by CCNE1 amplification, there is an urgent need to develop new therapeutic drugs targeting this amplification. PKMYT1 inhibitors as a promising treatment strategy can provide new treatment options for CCNE1 amplified tumors through a synthetic lethal approach.

[0005] Although a lot of efforts have been made in the development of Wee1 inhibitors and a number of clinical trials have been conducted, the development of PKMYT1 inhibitors is significantly lagging behind. Currently, only one PKMYT1 inhibitor candidate drug RP-6306 is undergoing phase II clinical trials. Research on PKMYT1 and the development of inhibitors still have important significance. Through in-depth study of the biological function of PKMYT1 and the development of inhibitors targeting it, new opportunities can be provided for cancer treatment, and the mechanism of action of PKMYT1 in cell cycle regulation and cancer progression can be further revealed. SUMMARY

[0006] The main purpose of the present application is to provide a new PKMYT1 small molecule inhibitor. In the first aspect of the present application, a compound of formula (I), a pharmaceutically acceptable salt, stereoisomer, isotopic derivative, solvate (such as hydrate, methanol solvate, ethanol solvate, propylene glycol solvate, isopropanol solvate, etc.) or prodrug thereof is provided,

[0007] wherein ring A is selected from a six-membered unsaturated nitrogen-containing heterocycle, which optionally contains one or two additional heteroatoms selected from N, O and S, and is optionally substituted with one or more R A , each independently selected from -D, -OH, halogen, oxo (=O), -CN, C1-10alkyl, C2-10alkenyl, C2-10alkynyl, 3-10 membered cycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl, -NHR1, -NHC(O)R1, -NHC(O)NHR1, or -NHSO2R1, and wherein R1is selected from -H, -D, -OH, -NH2, C1-10alkyl or 3-10 membered cycloalkyl; A , each independently selected from -D, -OH, halogen, oxo (=O), -CN, C1-10alkyl, C2-10alkenyl, C2-10alkynyl, 3-10 membered cycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl, -NHR1, -NHC(O)R1, -NHC(O)NHR1, or -NHSO2R1, and wherein R1is selected from -H, -D, -OH, -NH2, C1-10alkyl or 3-10 membered cycloalkyl;

[0008] Ring B is selected from a six-membered unsaturated nitrogen-containing heterocyclic ring, which optionally contains one or two additional heteroatoms selected from N, O and S, and is optionally substituted by one or more R B Substituted, where R B Each independently selected from -(C≡C) m -R2, -O-(CH2) n -R2, -SO2R2, -C(O)NR3R4, or -SO2NR3R4, wherein m is selected from 0 or 1, n is an integer selected from 0 to 3 (e.g., 0, 1, 2, 3), and wherein R2 is selected from -D, halogen, -CN, -OH, -NR3R4, optionally substituted by one or more R s1 Substituted C1-10 alkyl, optionally substituted by one or more R s1 Substituted C2-10 alkenyl, optionally substituted with one or more R s1 Substituted C2-10 alkynyl, optionally substituted by one or more R s1 Substituted C1-10 alkoxy, optionally substituted by one or more R s2 substituted 3-10 membered cycloalkyl, optionally substituted by one or more R s2 substituted 3-10 membered cycloalkenyl, optionally substituted with one or more R s2 substituted 3-10 membered heterocycloalkyl, optionally substituted by one or more R s2 substituted 3-10 membered heterocycloalkenyl, optionally substituted by one or more R s2 substituted 5-10 membered aryl, or optionally substituted with one or more R s2 substituted 5-10 membered heteroaryl, wherein R3 and R4 are each independently selected from -H, -D, -OH, -CN, optionally substituted by one or more R s1 Substituted C1-5 alkyl, optionally substituted by one or more R s1 Substituted C2-10 alkenyl, optionally substituted with one or more R s1 substituted 3-10 membered cycloalkyl, optionally substituted by one or more R s1 substituted 5-10 membered heteroaryl, -C(O)-R5 or -(CH2) p -O-R5, wherein p is an integer selected from 0 to 5 (e.g., 0, 1, 2, 3, 4, 5), wherein R5 is selected from -H, -D or C1-5 alkyl, R s1 are each independently selected from -D, -OH, halogen, -CN, -NH2, C1-5 alkyl, 3-10 membered cycloalkyl, C1-5 alkoxy or -C(O)-R5, and R s2 Each independently selected from -D, -OH, halogen, -CN, -NR3R4, oxo (=O), -C(O)-R5, -C(O)-NR3R4, optionally substituted by one or more Rs1 substituted C1-5alkyl, or optionally substituted 5-10 membered heteroaryl; s1 substituted 5-10 membered heterocycloalkyl;

[0009] Ring C is selected from optionally substituted phenyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolyl, or indazolyl; s1 substituted phenyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolyl, or indazolyl;

[0010] Q and U are each independently selected from N, C, or CH, and

[0011] represents a single or double bond.

[0012] In some embodiments, Ring A is selected from: and is optionally substituted with one or more R A substituted.

[0013] In some embodiments, Ring A is selected from: wherein R6, R7, R8are each independently selected from -H or R A , preferably wherein the carbon attached to U, and further preferably wherein R6, R7, R8are not simultaneously -H.

[0014] In some embodiments, R6is selected from -H, -D, -OH, halogen, -CN, C1-5alkyl, C2-6alkenyl, C2-6alkynyl, 3-8 membered cycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl, -NHR1, -NHC(O)R1, -NHC(O)NHR1, or -NHSO2R1, and wherein R1is selected from -H, -D, -OH, -NH2, C1-5alkyl, or 3-8 membered cycloalkyl. In some embodiments, R6is selected from -H, F, Cl, Br, -NH2, -NH-C1-5alkyl, -NHOH, -NHNH2, -NHCOH, or -NHC(O)-C1-5alkyl.

[0015] In some embodiments, R7is selected from -H, -D, -OH, halogen, -CN, C1-5alkyl, C2-6alkenyl, C2-6alkynyl, 3-8 membered cycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl, -NHR1, -NHC(O)R1, -NHC(O)NHR1, or -NHSO2R1, and wherein R1is selected from -H, -D, -OH, -NH2, C1-5alkyl, or 3-8 membered cycloalkyl. In some embodiments, R7is selected from -H, F, Cl, Br, -NH2, -NH-C1-5alkyl, or C1-5alkyl.

[0016] In some embodiments, R8 is selected from -H, -D, -OH, halogen, -CN, C1-5 alkyl, C2-6 alkenyl, C2-6 alkynyl, 5-10 membered aryl, 5-10 membered heteroaryl, -NHR1, -NHC(O)R1, -NHC(O)NHR1, or -NHSO2R1, and wherein R1 is selected from -H, -D, -OH, -NH2, and C1-5 alkyl. In some embodiments, R8 is selected from -H, F, Cl, Br, -NH2, -NH-C1-5 alkyl, or C1-5 alkyl.

[0017] In some embodiments, Ring A is selected from wherein the * carbon is connected to U, R6 is selected from F, Cl, Br, -NH2, -NH-C1-5 alkyl, -NHOH, -NHNH2, -NHCOH, or -NHC(O)-C1-5 alkyl, and / or R7 is selected from -H, F, Cl, Br, -NH2, -NH-C1-5 alkyl, or C1-5 alkyl. In some embodiments, ring A is selected from wherein the *carbon is connected to U, R6 is -NH2, and / or R7 is -H.

[0018] In some embodiments, Ring A is selected from Wherein * carbon is connected to U, R7 is selected from -H, F, Cl, Br, -NH2, -NH-C1-5 alkyl, or C1-5 alkyl. In some embodiments, ring A is selected from wherein the * carbon is connected to U, and R7 is selected from -H.

[0019] In some embodiments, Ring A is selected from Wherein the * carbon is connected to U, and R6 is selected from F, Cl, Br, -NH2, -NH-C1-5 alkyl, -NHOH, -NHNH2, -NHCOH, or -NHC(O)-C1-5 alkyl. In some embodiments, ring A is selected from wherein the * carbon is connected to U, and R6 is selected from -NH2.

[0020] In some embodiments, Ring A is selected from wherein the carbon atom is connected to U, R6 is selected from F, Cl, Br, -NH2, -NH-C1-5 alkyl, -NHOH, -NHNH2, -NHCOH, or -NHC(O)-C1-5 alkyl, and / or R7 is selected from hydrogen and deuterium. In some embodiments, ring A is selected from wherein the * carbon is connected to U, and R6 is selected from -NH2.

[0021] In some embodiments, Ring B is selected from: and optionally one or more R B replace.

[0022] In some embodiments, ring B is selected from wherein the *carbon is attached to U, and wherein R9, R 10 , R 11 are each independently selected from -H, -D, or R B . 10 , R 11 , preferably wherein any two of R9, R 10 , R 11 are not simultaneously -H.

[0023] In some embodiments, ring B is selected from wherein the *carbon is attached to U, and wherein R9, R 10 , R 11 are each independently selected from -H, -D, or R B .

[0024] In some embodiments, ring B is selected from wherein the *carbon is attached to U, and wherein R9and R 11 are -H, and R 10 is not -H and is selected from R B , preferably R 10 is not -H and is selected from -(CºC) m -R2, -0-(CH2) n -R2, or -S02R2, wherein m is selected from 0 or 1, n is selected from 0, 1, or 2, and wherein R2is selected from -H, -D, halogen (especially -Br), -CN, -OH, or optionally substituted with one or more -D, halogen (especially -F), -OH, or oxo: -NHCH3, -N(CH3)2, -NHCH2CH3, or -N(CH2CH3)2.

[0025] In some embodiments, ring B is selected from wherein the *carbon is attached to U, and wherein R9and R 10 are -H or -D, and R 11 is not -H and is selected from R B , preferably R 11 is not -H and is selected from -(CºC) m -R2, -0-(CH2) n -R2, or -S02R2, wherein m is selected from 0 or 1, n is selected from 0, 1, or 2, and wherein R2is selected from -H, -D, halogen (especially -Br), -CN, -OH, or optionally substituted with one or more -D, halogen (especially -F), -OH, or oxo: -N(CH3)2, -N(CH2CH3)2, -NHCH3, -NHCH2CH3, or -NHCH2CHF2.

[0026] In some embodiments, ring B is selected from wherein the *carbon is attached to U, and wherein R9is -H, and R 10 and R 11 are not -H and are each independently selected from R B , preferably, R 10 and R 11 are the same, also preferably, R 10 and R 11 are selected from -CH3, -CH2CH3, -CH2CH2CH3, -CF3, -CHF2, -CF2CF3, -CF2CF2CF3, -CD3, -CD2CD3, -CD2CD2CD3, or CN, also preferably, R 10 and R 11 are selected from -CH3.

[0027] In some embodiments, ring B is selected from wherein the *carbon is attached to U, and wherein R9is -H or -D, R 11 is selected from -CH3, -CH2CH3, -CH2CH2CH3, -CF3, -CHF2, -CF2CF3, -CF2CF2CF3, -CD3, -CD2CD3, -CD2CD2CD3, or CN, and R 10 is not -H and is selected from R B , preferably, R 10 is not -H and is selected from -(CºC) m -R2, -0-(CH2) n -R2, or -SO2R2, wherein m is selected from 0 or 1, n is selected from 0, 1, or 2, and wherein R2is selected from -H, -D, halogen (especially -Br), -CN, -OH, or -CH3, -CH2CH3, -CH2CH2CH3, -NHCH3, -N(CH3)2, -NHCH2CH3, or -N(CH2CH3)2, -NHCH(CH3)2, or -NHCH2CF3.

[0028] In some embodiments, ring B is selected from wherein the *carbon is attached to U, and wherein R 10 , R 11 are each independently selected from -H, -D, or R B, preferably wherein R 11 is -H and R 10 is not -H and is selected from R B , preferably wherein R 10 is not -H and is selected from -(CºC) m -R2, -0-(CH2) n -R2, or -S02R2, wherein m is selected from 0 or 1, n is selected from 0, 1 or 2, and wherein R2is selected from -H, -D, halogen (especially -Br), -CN, -OH or oxo optionally substituted with one or more -D, halogen (especially -F), -OH or oxo. (e.g. (e.g. (e.g. ), (e.g. ), -N(CH2CH3)2, -NHCH3, -NHCH2CH3, or -NHCH2CHF2.

[0029] In some embodiments, ring B is selected from wherein the *carbon is attached to U, and wherein R 10 is -H and R 11 is not -H and is selected from R B , preferably wherein R 11 is not -H and is selected from -(CºC) m -R2, -0-(CH2) n -R2, or -S02R2, wherein m is selected from 0 or 1, n is selected from 0, 1 or 2, and wherein R2is selected from -H, -D, halogen (especially -Br), -CN, -OH or oxo optionally substituted with one or more -D, halogen (especially -F), -OH or oxo.

[0030] In some embodiments, ring B is selected from wherein the *carbon is attached to U, and R 10 and R 11 are not -H and are each independently selected from R B , preferably R 10 and R 11 are the same, further preferably R 10 and R 11 are selected from -CH3, -CH2CH3, -CH2CH2CH3, -CF3, -CHF2, -CF2CF3, -CF2CF2CF3, -CD3, -CD2CD3, -CD2CD2CD3, or CN, further preferably R10 and R 11 Selected from -CH3.

[0031] In some embodiments, R9, R 10 、R 11 Each independently selected from -H, -(C≡C) m -R2, -O-(CH2) n -R2, -SO2R2, -C(O)NR3R4, or -SO2NR3R4, wherein m is selected from 0 or 1, n is an integer selected from 0 to 3 (e.g., 0, 1, 2, 3), and wherein R2 is selected from -D, halogen, -CN, -OH, -NR3R4, optionally substituted by one or more R s1 Substituted C1-5 alkyl, optionally substituted by one or more R s1 Substituted C2-6 alkenyl, optionally substituted by one or more R s1 Substituted C2-6 alkynyl, optionally substituted by one or more R s1 Substituted C1-5 alkoxy, optionally substituted by one or more R s2 substituted 3-8 membered cycloalkyl, optionally substituted by one or more R s2 substituted 3-8 membered cycloalkenyl, optionally substituted with one or more R s2 substituted 3-8 membered heterocycloalkyl, optionally substituted by one or more R s2 substituted 3-8 membered heterocycloalkenyl, optionally substituted by one or more R s2 substituted 5-10 membered aryl, or optionally substituted by one or more R s2 substituted 5-10 membered heteroaryl, wherein R3 and R4 are each independently selected from -H, -D, -OH, -CN, optionally substituted by one or more R s1 Substituted C1-5 alkyl, optionally substituted by one or more R s1 Substituted C2-6 alkenyl, optionally substituted by one or more R s1 substituted 3-8 membered cycloalkyl, optionally substituted by one or more R s1 substituted 5-10 membered heteroaryl, -C(O)-R5 or -(CH2) p -O-R5, wherein p is an integer selected from 0 to 5 (e.g., 0, 1, 2, 3, 4, 5), wherein R5 is selected from -H, -D or C1-5 alkyl, R s1 are each independently selected from -D, -OH, halogen, -CN, -NH2, C1-5 alkyl, 3-8 membered cycloalkyl or -C(O)-R5, and R s2 Each independently selected from -D, -OH, halogen, -CN, -NR3R4, oxo (=O), -C(O)-R5, -C(O)-NR3R4, optionally substituted by one or more R s1 substituted C1-5 alkyl, or optionally substituted by one or more Rs1 Substituted 5-10 membered heterocycloalkyl.

[0032] In some embodiments, R9 is selected from -H, -D, halogen, -CN, -OH, C1-5 alkyl optionally substituted with one or more halogen or -D, C2-6 alkenyl optionally substituted with one or more halogen or -D, or 3-8 membered cycloalkyl optionally substituted with one or more halogen or -D.

[0033] In some embodiments, R 10 and R 11 Each independently selected from -H, -(C≡C) m -R2, -O-(CH2) n -R2, or -SO2R2, wherein m is selected from 0 or 1, n is selected from 0, 1 or 2, and wherein R2 is selected from -H, -D, halogen, -CN, -OH, C1-5 alkyl optionally substituted with one or more halogen, -D, -OH, or -CN (e.g., substituted or unsubstituted methyl, ethyl or propyl, such as -CH2CF3, -CH2CHF2, -CF2CF3, -CH(CF3)2, -CF(CF3)2, -C(OH)(CF3)2, etc.), or C2-6 alkenyl (e.g., optionally substituted with one or more halogen, -D, -OH, or -CN) ), or C2-6 alkynyl optionally substituted by one or more halogen, -D, -OH, or -CN, or C1-5 alkoxy optionally substituted by one or more halogen, -D, -OH, or -CN (e.g. ).

[0034] In some embodiments, R 10 and R 11 Each independently selected from: -NR3R4, -O-(CH2) n -NR3R4, -C(O)NR3R4, or -SO2NR3R4, wherein n is an integer selected from 0 to 3 (e.g., 0, 1, 2, 3), and wherein R3 and R4 are each independently selected from -H, -D, -OH, -CN, optionally substituted by one or more R s1 Substituted C1-5 alkyl, optionally substituted by one or more R s1 Substituted C2-6 alkenyl, optionally substituted by one or more R s1 substituted 3-8 membered cycloalkyl, optionally substituted by one or more R s1 substituted 5-10 membered heteroaryl, -C(O)-R5 or -(CH2) p -O-R5, wherein p is an integer selected from 0 to 5, R5 is selected from -H, -D or C1-5 alkyl, and wherein R s1each independently selected from -D, -OH, halogen (especially F), or -CN.

[0035] In some embodiments, the substituent -NR3R4(including the moiety of -O- n -NR3R4, -C(O)NR3R4, or -SO2NR3R4) is selected from -NHCH3, -NHCH2CH3, -NHCH2CH2CH3, -NHCH(CH3)2, -N(CH3)2, -N(CH2CH3)2, -N(CH2CH2CH3)2, -NHCH2CH2OCH3, -NHCH=CH2, -NHCH2CH=CH2, -NHCH=CHCH3, -N(CH=CH2)2, -N(CH2CH=CH2)2, -N(CH=CHCH3)2,

[0036] In some embodiments, R 10 and R 11 each independently selected from -H, -(CºC) m -R2, -O-(CH2) n -R2, or -SO2R2, wherein m is selected from 0 or 1, n is selected from 0, 1, or 2, and wherein R2is selected from: wherein q is an integer independently selected from 0-6 (e.g., 0, 1, 2, 3, 4, 5, 6), and R s2 as defined above.

[0037] In some embodiments, R 10 and R 11 each independently selected from -H, -(CºC) m -R2, -O-(CH2) n -R2, or -SO2R2, wherein m is selected from 0 or 1, n is selected from 0, 1, or 2, and wherein R2is selected from:

[0038] In some embodiments, ring C is selected from wherein R s1each independently selected from -D, -OH, halogen, -CN, -NH2, C1-5alkyl, 3-10 membered cycloalkyl, C1-5alkoxy, or -C(O)-R5, wherein R5is selected from -H, -D, or C1-5alkyl, and q is an integer from 0 to 5 (e.g., 0, 1, 2, 3, 4, 5), or a stereoisomer, tautomer, prodrug, or pharmaceutically acceptable salt of said compound.

[0039] In some embodiments, ring C is selected from wherein R 12 , R 13 , R 14 , R 15 each independently selected from -H, -D, -OH, halogen, -CN, -NH2, C1-5alkyl, or -C(O)-R5, wherein R5is selected from -H, -D, or C1-5alkyl.

[0040] In some embodiments, ring C is selected from

[0041] In some embodiments, the compound is of Formula (IIA), Formula (IIB), Formula (IIC), or Formula (IID):

[0042] wherein R6, R7, R9, R 10 , R 11 , R 12 , R 13 , R 14 , R 15 are as defined above.

[0043] In some embodiments, the compound is enriched for the atropisomer of Formula (IIAi), Formula (IIBi), Formula (IICi), or Formula (IIDi):

[0044] wherein R6, R7, R9, R 10 , R 11 , R 12 , R 13 , R 14 , R 15 are as defined above.

[0045] In some embodiments, the compound is enriched for the atropisomer of Formula (IIAii), Formula (IIBii), Formula (IICii), or Formula (IIDii):

[0046] wherein R6, R7, R9, R 10 , R 11 , R 12 , R 13 are as defined above.

[0047] In some embodiments, the compound has Formula (IIA-1) or Formula (IIA-2):

[0048] in,

[0049] R7 is hydrogen or deuterium;

[0050] R 15 is hydrogen or halogen (preferably, F);

[0051] R 11 is hydrogen, CN, or methyl or ethyl optionally substituted by 1 or more (e.g. 1, 2, 3, 4 or 5) deuterium;

[0052] R 10 Selected from:

[0053] - halogen (such as F, Cl or Br, preferably Br),

[0054] - optionally substituted by 1 or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) R k Substituted C 2-3 Alkyl (such as ethyl, n-propyl or isopropyl), C 2-3 alkenyl (e.g., vinyl, propenyl, allyl, isopropenyl), cyclopropyl, 5-6 membered heterocycloalkyl containing 1 or 2 heteroatoms independently selected from O or N, 5-6 membered heterocycloalkenyl containing 1 or 2 heteroatoms independently selected from O or N, 5-6 membered heteroaryl containing 1 or 2 heteroatoms independently selected from O, N or S, wherein R k are each independently selected from halogen (preferably, F or Cl), hydroxyl, cyano or deuterium,

[0055] - unsubstituted phenyl, or

[0056] --NR3R4, wherein R3 and R4 are each independently selected from H, C optionally substituted with 1 to 7 (e.g., 1, 2, 3, 4, 5, 6 or 7) deuterium 1-3 Alkyl or cyclopropyl;

[0057] The conditions are:

[0058] i) the heterocycloalkenyl group contains only one double bond;

[0059] ii) When R 11 When it is CN, R 10 Selected from unsubstituted C 2-3 Alkyl or unsubstituted C 2-3 alkenyl;

[0060] iii) When R 10when R is -NR3R4, R3and R4are not both C

[0061] iv) when R 10 is -NR3R4, R3and R4are not both C 2-3 alkyl, or cyclopropyl optionally substituted with deuterium, or C 2-3 alkyl and cyclopropyl;

[0062] v) when R 10 is a 5-6 membered heterocycloalkyl containing 1 N atom, which is attached to the pyridine ring of formula (IIA-1) or formula (IIA-2) through the N atom; or

[0063] vi) when R 10 is a 5-6 membered heteroaryl containing 1 N atom, which is not attached to the pyridine ring of formula (IIA-1) or formula (IIA-2) through the N atom.

[0064] In some embodiments, the 5-6 membered heterocycloalkyl containing 1 heteroatom selected from O or N is selected from:

[0065] In some embodiments, the 5-6 membered heterocycloalkenyl containing 1 heteroatom selected from O or N is selected from:

[0066] In some embodiments, the 5-6 membered heteroaryl containing 1-2 heteroatoms independently selected from O, N or S is selected from:

[0067] In some embodiments, when R 11 is hydrogen or methyl optionally substituted with 1-3 (e.g., 1, 2, or 3) deuterium, R 10 is selected from:

[0068] - F, Cl, Br,

[0069] - ethyl optionally substituted with 1 or more (e.g., 1, 2, 3, 4, 5, 6, or 7) R k substituted with 1 or more (e.g., 1, 2, 3, 4, 5, 6, or 7) R

[0070] - ethyl optionally substituted with 1 or more (e.g., 1, 2, 3, 4, 5, 6, or 7) R ksubstituted tetrahydrofuranyl, tetrahydropyrrolyl, tetrahydropyranyl, piperidinyl,

[0071] - optionally substituted by 1 or more (e.g. 1, 2, 3, 4, 5, 6 or 7) R k substituted

[0072] - optionally substituted by 1 or more (e.g. 1, 2, 3 or 4) R k substituted pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, isoxazolyl, pyrazolyl, thiazolyl, pyridinyl, for example

[0073] - unsubstituted phenyl, or

[0074] - NR3R4, wherein R3and R4are each independently selected from H, methyl, ethyl, n-propyl, i-propyl or cyclopropyl optionally substituted with 1 to 7 deuterium.

[0075] -

[0076] In some embodiments, when R 11 is hydrogen or methyl optionally substituted with 1-3 deuterium, R 10 is selected from:

[0077] - Br,

[0078] - ethyl, i-propyl, vinyl, propenyl, allyl, i-propenyl, cyclopropyl, substituted with 1 to 7 R k substituted ethyl, i-propyl, vinyl, propenyl, allyl, i-propenyl, cyclopropyl,

[0079] - unsubstituted ethyl, i-propyl, vinyl, propenyl, allyl, i-propenyl, cyclopropyl,

[0080] - unsubstituted tetrahydrofuranyl, tetrahydropyrrolyl, tetrahydropyranyl, piperidinyl,

[0081] - unsubstituted

[0082] - unsubstituted pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, isoxazolyl, pyrazolyl, thiazolyl, pyridinyl, for example

[0083] - unsubstituted phenyl, or

[0084] --NR3R4, wherein R3 and R4 are each independently selected from H, methyl, ethyl, isopropyl or cyclopropyl optionally substituted with 1 to 5 deuteriums.

[0085] In some embodiments, when R 11 When R is an ethyl group optionally substituted with 1 to 5 deuterium groups, 10 are selected from optionally 1 to 4 (e.g. 1, 2, 3 or 4) R k Substituted pyrrolyl, furyl, thienyl, imidazolyl, oxazolyl, isoxazolyl, pyrazolyl, thiazolyl (e.g. ), or optionally 1 to 7 R k Substituted isopropyl or isopropenyl, wherein R k Each is independently selected from halogen (preferably, F or Cl) or deuterium (e.g., -C(=CH2)(CH3), -CH(CH3)2, -C(=CH2)(CD3), -CH(CH3)(CD3), -C(=CH2)(CF3) or -CH(CH3)(CF3)).

[0086] In some embodiments, the compound has Formula (IIB-1) or Formula (IIB-2):

[0087] in,

[0088] R 10 Selected from:

[0089] - optionally substituted by 1 to 6 R each independently selected from halogen (preferably F or Cl), hydroxyl, cyano or deuterium k Substituted isopropyl, vinyl, propenyl, allyl, isopropenyl, cyclopropyl,

[0090] - optionally substituted by 1 to 2 R each independently selected from halogen, deuterium, hydroxyl or methyl k substituted 5-6 membered heterocycloalkyl containing 1 heteroatom selected from O or N (e.g. tetrahydrofuranyl, tetrahydropyrrolyl, tetrahydropyranyl, piperidinyl),

[0091] - optionally substituted by 1 to 2 R each independently selected from halogen, deuterium, hydroxyl or methyl k substituted 5-6 membered heteroaryl containing 1 heteroatom selected from O, N or S, such as pyrrolyl, furyl, thienyl, imidazolyl, oxazolyl, isoxazolyl, pyrazolyl, thiazolyl, pyridyl, or

[0092] - unsubstituted phenyl;

[0093] The conditions are:

[0094] i) When R 10In some embodiments, the 5-6 membered heteroaryl containing 1 heteroatom selected from O, N, or S is selected from:

[0095] In some embodiments, R 10 is selected from isopropyl, -CH(CH3)(CD3), -CH(CH3)(CF3), -CF(CH3)2, or -C(OH)(CH3)2.

[0096] In some embodiments, the 5-6 membered heterocycloalkyl containing 1 heteroatom selected from O or N is optionally substituted is selected from:

[0097] In some embodiments, the 5-6 membered heteroaryl containing 1 heteroatom selected from O, N, or S is selected from:

[0098] In some embodiments, the compound is of formula (IID-1) or formula (IID-2):

[0099] wherein,

[0100] R7and R8are each independently hydrogen or deuterium;

[0101] R 10 is C 2-3 alkyl optionally substituted with halogen (preferably, F) or deuterium.

[0102] As used in the present application, “enriched” refers to a composition containing a greater proportion or percentage of one stereoisomer of a compound of the present application relative to other stereoisomer(s), e.g., it can have an enantiomeric excess ee higher than 80%, preferably higher than 90%, even higher than 95%, 96%, 97%, or 98%.

[0103] Accordingly, in some embodiments, the present application also provides a compound of formula (IIA),, formula (IIA-1), formula (IIB), formula (IIB-1) formula (IIC), formula (IID), or formula (IID-1). In some embodiments, the present application also provides a compound of formula (IIAi), formula (IIBi), formula (IICi), or formula (IIDi). In some embodiments, the present application also provides a compound of formula (IIAii), formula (IIA-2), formula (IIBii), formula (IIB-2), formula (IICii), formula (IIDii), or formula (IID-2).

[0104] In some embodiments, the compound is selected from the group consisting of the compounds in Table 1.

[0105] Table 1 : Exemplary compounds of the application

[0106] In a second aspect of the application, there is provided a pharmaceutical composition comprising a compound according to the first aspect of the application, or a pharmaceutically acceptable salt, stereoisomer, isotopic derivative, solvate or prodrug thereof, and a pharmaceutically acceptable carrier or excipient.

[0107] In some embodiments, the composition is enriched in the deuterium isotope.

[0108] In a third aspect of the application, there is provided use of a compound according to the first aspect of the application, or a pharmaceutically acceptable salt, stereoisomer, isotopic derivative, solvate or prodrug thereof, in the manufacture of a medicament for inhibiting the activity of PKMYT1 in a cell.

[0109] In some embodiments, the medicament is for treating cancer. In some embodiments, PKMYT1 is overexpressed in the cancer.

[0110] In a fourth aspect of the application, there is provided a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound according to the first aspect of the application, or a pharmaceutically acceptable salt, stereoisomer, isotopic derivative, solvate or prodrug thereof, or a pharmaceutical composition according to the second aspect of the application.

[0111] In some embodiments, PKMYT1 is overexpressed in the cancer.

[0112] In some embodiments, the cancer is selected from breast cancer, ovarian cancer, uterine cancer, gastric cancer, lung cancer (especially non-small cell lung cancer), hepatocellular carcinoma, glioblastoma, neuroblastoma, colorectal cancer, or esophageal cancer.

[0113] In a fifth aspect of the application, there is provided a method of inhibiting the activity of PKMYT1 in a cell overexpressing the same, the method comprising contacting the cell with a compound according to the first aspect of the application, or a pharmaceutically acceptable salt, stereoisomer, isotopic derivative, solvate or prodrug thereof, or a pharmaceutical composition according to the second aspect of the application.

[0114] In some embodiments, the cell is a mammalian cell. In some embodiments, the method is an in vitro non-therapeutic method. In some embodiments, the cell is in vivo in a subject.

[0115] In some embodiments, the mammal is a human.

[0116] The present application proves the high efficiency, safety and feasibility of the compounds in the present application in targeted therapy through multi-dimensional verification of enzyme activity inhibition, cytotoxicity, safety and in vivo efficacy, and the compounds in the present application have strong PKMYT1 inhibiting effect and can be used as drugs for treating PKMYT1 overexpression related diseases. DETAILED DESCRIPTION

[0117] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The following definitions are applied to the descriptions of the present application and throughout this document, and apply equally, where appropriate, to the singular as well as the plural versions of the terms.

[0118] Definitions

[0119] As described herein, the compounds in the present application can be substituted with any number of substituents or functional groups to expand its scope. Generally, the term "substituted" as used herein when used in the context of a general formula including substituents in the present application, whether preceded by the term "optionally" or not, means the replacement of hydrogen radicals with the designated structural substituents. When multiple positions in a particular structure are substituted with multiple particular substituents, the substituents can be the same or different at each position. The term "substituted" as used herein includes all permissible organic compound substitutions. Broadly, permissible substituents include acyclic, cyclic, branched, unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic organic compounds. In the present application, as in heteroatom nitrogen, it can have a hydrogen substituent or any permissible substituent to complement its valence. In addition, the present application considers the combination of substituents and variable groups to provide a stable form of the compound, thereby facilitating the treatment of diseases. The term "stable" here refers to a compound with a stable structure, which is detected for a long enough time to maintain the integrity of the compound structure, preferably for a long enough time to be effective.

[0120] Unless otherwise specified, the writing of a substituent does not constitute a limitation on the manner in which it is attached, for example, when ring A is selected from the substituent It can be connected to U in formula (I) through the left arm of the substituent, or through the right arm of the substituent.

[0121] The term "alkyl" refers to straight or branched chain saturated aliphatic hydrocarbon groups having the indicated number of carbon atoms. Alkyl groups used in the present application, including where they are part of another substituent, can contain 1 to 20 carbon atoms, for example 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 carbon atoms, preferably 1 to 15 carbon atoms, more preferably 1 to 10 carbon atoms, further preferably 1 to 5 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl, t-butyl, n-pentyl, i-pentyl, neopentyl, n-hexyl and various branched isomers thereof.

[0122] The term "alkenyl" refers to straight or branched chain, acyclic unsaturated hydrocarbon groups having the indicated number of carbon atoms, wherein at least two carbon atoms are bonded to one another by an unsaturated double bond. Alkenyl groups suitable for use in the present application can have 2-10 (e.g. 2, 3, 4, 5, 6, 7, 8, 9, 10) carbon atoms, preferably 2-5 carbon atoms. Examples of C2-5alkenyl groups include, but are not limited to, ethenyl, 1 -propen-1 -yl, 1 -propen-2-yl, 2-propen-1 -yl, 2-methyl-1 -propen-1 -yl, 2-methyl-2-propen-1 -yl, 1 -buten-1 -yl, 1 -buten-2-yl, 2-buten-1 -yl, 2-buten-2-yl, 3-buten-1 -yl, 3-buten-2-yl, 1,3-budien-1 -yl, 1,3-budien-2-yl, 1 -penten-1 -yl, 2-penten-1 -yl, 2-penten-2-yl, 3-penten-1 -yl, 3-penten-3-yl, 4-penten-1 -yl, 4-penten-4-yl and the like. The alkenyl group preferably has one double bond. It is also preferred that the double bond in the alkenyl group is directly attached to the remainder of the compound containing the alkenyl group, e.g. 1 -propen-1 -yl is preferred over 2-propen-1 -yl, whereby it is possible to form an extended conjugated system when the substituent is attached to ring A or ring B.

[0123] The term "alkynyl" refers to straight or branched chain, acyclic unsaturated hydrocarbon groups having the indicated number of carbon atoms, wherein at least two carbon atoms are bonded to one another by an unsaturated triple bond. Alkynyl groups suitable for use in the present application can have 2-10 (e.g. 2, 3, 4, 5, 6, 7, 8, 9, 10) carbon atoms, preferably 2-5 carbon atoms. The alkynyl group preferably has one triple bond. It is also preferred that the triple bond in the alkynyl group is directly attached to the remainder of the compound containing the alkynyl group.

[0124] The term "aryl" refers to a monocyclic, bicyclic, or tricyclic ring system (preferably a 5-10 membered aromatic ring) having a total of 5 to 15 ring members (or ring atoms), wherein all rings in the system are aromatic and wherein each ring in the system contains 3 to 7 ring members; when not all rings in the system are aromatic, it is considered to be a cycloalkyl or heterocycloalkyl. In certain embodiments of the application, "aryl" refers to an aromatic ring system, which includes, but is not limited to, phenyl, biphenyl, indanyl, 1-naphthyl, 2-naphthyl, tetrahydronaphthyl, anthryl. A fused aryl group can be attached to another group at a suitable position on a cycloalkyl ring or an aromatic ring.

[0125] The term "heteroaromatic ring" or "heteroaryl" refers to a heterocycle that is aromatic. Non-limiting examples of heteroaryl groups include thienyl, furanyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, triazolyl, thiazolyl, pyridyl, pyrazinyl, pyridazinyl, pyrimidinyl, indolyl, quinolinyl, isoquinolinyl, quinazolinyl, indazolyl, benzofuranyl, benzothienyl, benzoxazolyl, benzotriazolyl, or benzothiazolyl.

[0126] The term "cycloalkyl" refers to a cyclic alkyl group including saturated monocyclic, bicyclic, or polycyclic rings, such as 3-10 membered cycloalkyl groups, such as monocyclic, bicyclic, or polycyclic rings having 3, 4, 5, 6, 7, 8, 9, 10 carbon ring atoms. Cycloalkyl groups can also include cycloalkyl groups having spiro, bridged, annelated, and the like structures, such as 5-10 membered fused bicyclic rings, 5-10 membered spiro bicyclic rings, and the like. Cycloalkyl groups suitable for use in the present application include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and norbornyl. Examples of 5-10 membered fused bicyclic rings include, but are not limited to, Examples of 5-10 membered spiro bicyclic rings include, but are not limited to, Examples of 5-10 membered spiro bicyclic rings include, but are not limited to,

[0127] The term "cycloalkenyl" refers to a cycloalkyl group containing one or more unsaturated double bonds.

[0128] The term "heterocycloalkyl" refers to substituted or unsubstituted non-aromatic, partially unsaturated or fully saturated heterocyclyl groups. Heterocyclyl groups can contain 1 to 3 heteroatoms selected from N, O or S as ring members and can include monocyclic heterocycles, bicyclic bridged heterocycles, bicyclic fused heterocycles and bicyclic spiro heterocycles, etc. Heterocyclyl groups suitable for use in the present application can contain 3 to 12 ring atoms (3-12 membered heterocyclyl), for example 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 ring atoms, preferably 3-12, more preferably 3-10, and even more preferably 3-8 ring atoms. Heterocyclyl groups can be attached at a heteroatom or a carbon atom. The N, S in the heterocyclyl ring can be oxidized to various oxidation states. Non-limiting examples of heterocyclyl groups include oxetanyl, azetidinyl, tetrahydrofuranyl, tetrahydropyrrolyl, tetrahydrothiophenyl, tetrahydropyranyl, tetrahydrothiopyranyl, dioxanyl, piperazinyl, hexahydropyrazinyl, morpholinyl, piperidinyl, dithianyl, etc., for example

[0129] The term "heterocycloalkenyl" refers to a heterocycloalkyl group containing one or more unsaturated double bonds.

[0130] The term "six-membered unsaturated nitrogen-containing heterocycle" refers to a carbocycle containing one or more unsaturated double bonds, having 6 ring atoms and one of the ring atoms being an N heteroatom. The six-membered unsaturated nitrogen-containing heterocycle can also contain one or two additional heteroatoms selected from N, O and S, and thus, for example, when it contains one additional N heteroatom, two of the ring atoms in such a heterocycle are N heteroatoms.

[0131] The term "alkoxy" or "alkyloxy" refers to -O-alkyl, for example -O-C1-C5alkyl, more preferably -O-C1-C3alkyl. Non-limiting examples of alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, isobutoxy, t-butoxy, n-pentoxy, n-hexoxy, cyclopropyloxy and cyclobutyloxy, etc. 10 The term "alkyl" refers to a saturated straight-chain or branched-chain monovalent hydrocarbon group having from 1 to 10 carbon atoms, preferably 1 to 5 carbon atoms, more preferably 1 to 3 carbon atoms. Non-limiting examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, t-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, and the like. The term "alkyl" also includes cycloalkyl groups, which are saturated cyclic alkyl groups, and the like.

[0132] The term "halogen" refers to F, Cl, Br, I, or isotopes thereof.

[0133] The term "halo" or "halogen substitution" refers to substitution of a hydrogen atom by one or more halogen atoms selected from F, Cl, Br, I, or isotopes thereof, the upper limit of the number of halogen substituents being equal to the sum of the number of hydrogens that the substituted group can have, and in the absence of a specific limitation, the number of halogen substituents is any integer between 1 and the upper limit, and when the number of halogen substituents is greater than 1, the halogen substituents can be the same or different halogen.

[0134] The term "deuterated" or "deuterated form" as used herein refers to the substitution of at least one isotope of deuterium for a hydrogen atom on an alkyl, cycloalkyl, aryl, heteroaryl, heterocycloalkyl, alkenyl, alkynyl, and the like, the upper limit of deuterium substitution being equal to the sum of hydrogens that the substituted group can have. In the absence of a specific limitation, the number of deuterium substitutions is any integer between 1 and the upper limit, preferably 1-10 deuterium substitutions, more preferably 1-6 deuterium substitutions, and even more preferably 1-3 deuterium substitutions. In the absence of a specific limitation, the compounds of the present application encompass instances in which one or more hydrogen atoms are substituted with the isotope deuterium.

[0135] In the present application, "oxo" refers to a double-bonded oxygen (=0).

[0136] A line drawn from a ring system indicates that one end of the bond can be connected to any suitable ring atom in the ring system through which the bond is drawn, through one ring, indicating connection at any optional position on that ring, through multiple rings, indicating connection at any optional position on the multiple rings. For example, the structure of includes any one of the following structures:

[0137] The term "treat" a disease in a subject or "treat" a subject having or suspected of having a disease refers to administering a drug treatment to the subject, e.g., administering one or more agents, so as to reduce or prevent worsening of at least one symptom of the disease. Thus, in one embodiment, "treat" refers, inter alia, to delaying progression, accelerating remission, inducing remission, increasing remission, accelerating recovery, increasing efficacy of or reducing resistance to alternative therapies, or combinations thereof. For example, reducing or maintaining the number or volume of detectable tumors, or reducing their invasiveness and metastatic ability, or reducing their resistance to known therapies or chemotherapies, etc., in a treated patient population relative to an untreated control population.

[0138] In some embodiments, a "therapeutically effective amount" refers to an amount of a compound disclosed herein that, when administered, will relieve, to some extent, one or more of the symptoms of the disease or condition being treated. In some embodiments, the result is a decrease and / or alleviation of a sign, symptom, or cause of disease, or any other desired alteration of a biological system. For example, an "effective amount" for therapeutic uses is the amount of a composition comprising a compound disclosed herein that will provide a clinically significant decrease in disease symptoms.

[0139] The term "cancer" can include cancers that are the result of genetic hereditary mutations. Examples of such cancers include, but are not limited to, breast cancer, cancers associated with Li-Fraumeni syndrome, e.g., childhood sarcomas, leukemias, and brain cancers, cancers associated with Lynch syndrome, e.g., colon cancer, bile duct cancer, brain cancer, endometrial cancer, kidney cancer, ovarian cancer, pancreatic cancer, small bowel cancer, stomach cancer, and ureter cancer, lung cancer, melanoma, prostate cancer, retinoblastoma, thyroid cancer, and uterine cancer. In addition, cancers can be the result of acquired mutations, e.g., diet, environmental, and / or lifestyle induced mutations or somatic mutations. Examples of such cancers can include, but are not limited to, adrenal cancer, adrenocortical cancer, bladder cancer, brain cancer, primary brain cancer, glioma, glioblastoma, breast cancer, cervical cancer, colon cancer (non-limiting examples include colorectal cancers such as colon adenocarcinoma and colon cancer), endometrial cancer, epidermal cancer, esophageal cancer, gallbladder cancer, genitourinary tract cancer, head and neck cancer, kidney cancer, liver cancer, lung cancer (non-limiting examples include adenocarcinoma, small cell lung cancer, and non-small cell lung cancer), lymphoma (non-limiting examples include B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma), melanoma, malignant melanoma, malignant carcinoid, malignant pancreatic insulinoma, myeloma, multiple myeloma, ovarian cancer, pancreatic cancer (e.g., exocrine pancreatic cancer), prostate cancer, renal cell carcinoma, skin cancer, e.g., squamous cell carcinoma, stomach cancer, testicular cancer, thyroid cancer, thyroid follicular cancer, Wilms' tumor, choriocarcinoma, mycosis, malignant hypercalcemia, cervical hyperplasia, leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, hairy cell lymphoma, Burkitt's lymphoma, acute myelocytic leukemia, chronic myelocytic leukemia, myelodysplastic syndrome, promyelocytic leukemia, chronic myelogenous leukemia, acute myelogenous leukemia, fibrosarcoma, habdomyosarcoma, astrocytoma, neuroblastoma, rhabdomyosarcoma, schwannoma, Kaposi's sarcoma, polycythemia vera, essential thrombocythemia, Hodgkin's disease, non-Hodgkin's lymphoma, soft-tissue sarcoma, osteogenic sarcoma, primary macroglobulinemia, seminoma, teratoma, osteosarcoma, xenoderma pigmentosum, keratoacanthoma, and retinoblastoma.

[0140] The term "subject" refers to an animal, such as a mammal, including, but not limited to, primates (e.g., humans), cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice, and the like. In some embodiments, the subject is a human.

[0141] The term "pharmaceutical composition" refers to a mixture of one or more compounds of the application or pharmaceutically acceptable salts, stereoisomers, isotopically labeled derivatives, solvates, or prodrugs thereof with other components, such as physiologically / pharmaceutically acceptable carriers and excipients.

[0142] The term "pharmaceutically acceptable salt" refers to (i) salts of acidic groups present in the compounds provided herein with appropriate inorganic or organic cations (bases), and includes, but is not limited to, alkali metal salts, such as sodium salts, potassium salts, lithium salts, and the like; alkaline earth metal salts, such as calcium salts, magnesium salts, and the like; other metal salts, such as aluminum salts, iron salts, zinc salts, copper salts, nickel salts, cobalt salts, and the like; inorganic base salts, such as ammonium salts; organic base salts, such as tertiary octylamine salts, dibenzylamine salts, morpholine salts, glucosamine salts, phenylglycine alkyl ester salts, ethylenediamine salts, N-methylglucamine salts, guanidine salts, diethylamine salts, triethylamine salts, dicyclohexylamine salts, N,N'-dibenzylethylenediamine salts, chloroprocaine salts, procaine salts, diethanolamine salts, N-benzyl-phenethylamine salts, piperazine salts, tetramethylamine salts, tris(hydroxymethyl)aminomethane salts, and (ii) salts of basic groups present in the compounds provided herein with appropriate inorganic or organic anions (acids), and includes, but is not limited to, hydrogen halide salts, such as hydrofluoride salts, hydrochloride salts, hydrobromide salts, hydroiodide salts, and the like; inorganic acid salts, such as nitrate salts, perchlorate salts, sulfate salts, phosphate salts, and the like; lower alkyl sulfonic acid salts, such as methanesulfonate salts, trifluoromethanesulfonate salts, ethanesulfonate salts, and the like; arylsulfonic acid salts, such as benzenesulfonate salts, p-toluenesulfonate salts, and the like; organic acid salts, such as acetate salts, malate salts, fumarate salts, succinate salts, citrate salts, tartrate salts, oxalate salts, maleate salts, and the like; amino acid salts, such as glycine salts, trimethylglycine salts, arginine salts, ornithine salts, glutamic acid salts, aspartic acid salts, and the like. See, Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl, 2002).

[0143] The compounds or salts in the present application can exist in enantiomeric and diastereomeric forms produced by an axis of chirality, chiral center. All such stereoisomers are part of the present application. The term "stereoisomers" refers to compounds which have identical chemical constitution, but differ with regard to the arrangement of the atoms or groups in space. The individual stereoisomers of the compounds in the present application can not be present in combination with other isomers (e.g., as a particular enantiomer having an extraordinary activity), or can be present in mixtures, such as racemates, or mixtures with all other stereoisomers, or a portion thereof. Thus, the compounds of the present application include all stereoisomers, including diastereomeric, atropisomeric, enantiomeric, mixtures thereof and racemic mixtures of the compounds of Formula (I), Formula (IIA), Formula (IIB) or Formula (IIC), including pharmaceutically acceptable salts thereof. When one isomer (e.g., enantiomer, diastereomer, atropisomer or geometric isomer) is inherently more active as an inhibitor of PKMYT1 than its opposite isomer, the more active isomer is preferred. Ring C in the compounds of the present application can exhibit conformational rotational isomerism, referred to herein as atropisomers.

[0144] The term "atropisomer" refers to stereoisomers that result from restricted rotation about a single bond, where the rotational barrier is high enough to allow separation of the isomers. Typically, rotation about a single bond in a molecule is prevented or greatly slowed due to steric interactions with other parts of the molecule and the substituents on either side of the single bond are not symmetric, resulting in the formation of a stereo unit called a "chiral axis".

[0145] The present application also contemplates tautomers of the compounds described and pharmaceutically acceptable salts, solvates and prodrugs thereof. "Tautomers" refer to compounds whose structures differ significantly in terms of the arrangement of atoms but exist in an easy and rapid equilibrium. Common tautomeric pairs are: keto-enol, amide-nitrile, lactam-lactim, amide-imidic acid tautomerism in heterocycles (e.g., in the nucleobases guanine, thymine and cytosine), amine-enamine and enamine-enamine.

[0146] The term "isotopically-labelled" refers to the substitution of one or more atoms in a compound with an atom having an atomic mass or mass number that differs from the atomic mass or mass number most commonly found in nature. The term "isotopically-labelled" also refers to a compound that includes one or more atoms that is an atom having an atomic mass or mass number that differs from the atomic mass or mass number most commonly found in nature. Examples of isotopes that can be labelled in the compounds of the present application include hydrogen, carbon, nitrogen, oxygen, phosphorus, sulphur, fluorine and chlorine isotopes, such as 2 H, 3 H, 13 C, 11 C, 14 C, 15 N, 18 O, 17 O, 31 P, 32 P, 35 S, 18 F and36 In some embodiments, heavier isotopes such as deuterium are substituted, i.e. 2 H or D, due to their excellent metabolic stability, have advantages in certain therapies, such as increasing half-life in vivo or reducing dosage, and therefore, may be preferred in certain circumstances. Isotope-labeled compounds can be prepared using conventional methods by replacing readily available isotope-labeled reagents with non-isotopic reagents, using the protocols disclosed in the Examples.

[0147] The term "prodrug" refers to a compound that undergoes chemical conversion by metabolic or chemical processes to produce the compound of the present invention or a salt thereof when used to treat a related disease.

[0148] The pharmaceutical composition of the present invention may contain pharmaceutically acceptable excipients, including but not limited to: ion exchangers, aluminum oxide, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphates, glycerol, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulosic substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, beeswax, lanolin, etc.

[0149] "Pharmaceutically acceptable carrier" refers to one or more compatible solid or liquid fillers or gel substances that are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0150] A "pharmaceutically acceptable excipient" refers to a substance that aids in the formulation and / or administration of an active agent and / or is absorbed by the individual, and can be included in the compositions of the present disclosure without causing a significant adverse toxicological effect on the individual. Non-limiting examples of pharmaceutically acceptable carriers and excipients include water, NaCl, normal saline solutions, lactated Ringer's solution, normal sucrose, normal glucose, binding agents, fillers, disintegrating agents, lubricants, coatings, sweetening, flavoring, salt solutions (such as Ringer's solution), alcohols, oils, gelatins, carbohydrates such as lactose, amylose or starch, fatty acids esters, hyrmethylcellulose, polyvinyl pyrrolidine and pigments and the like. Such preparations can be sterilized and, if desired, mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, and / or aromatic substances and the like that do not deleteriously react with the compounds provided herein or interfere with their activity. Those of ordinary skill in the art will recognize additional pharmaceutical carriers and excipients suitable for use in the disclosed compounds.

[0151] In certain embodiments, the pharmaceutical compositions of the present application can be in solid or liquid form.

[0152] The pharmaceuticals containing the active ingredient can be in a form suitable for oral administration as suitable tablets, pills, capsules, aqueous or oily suspensions, dispersible powders or granules, syrups or elixirs, for example. The pharmaceuticals for oral use are produced according to known methods in the pharmaceutical art, and these compositions can include one or more agents such as sweetening, flavoring, coloring agents and preserving agents in order to provide a palatable or tasty pharmaceutical preparation. The tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets. Examples of such excipients are inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, such as corn starch, or alginic acid; binding agents, such as starch, gelatin or acacia; and lubricating agents such as magnesium stearate, stearic acid or talc. The tablets can be uncoated or they can be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over an extended period of time. Examples of coating agents include polymeric substances such as hydroxypropylmethylcellulose, shellac, and the like.

[0153] The pharmaceutical compositions of the present application can be prepared according to different routes of administration in various forms. For example, the pharmaceutical compositions can be administered in any of the following ways: orally, spray inhalation, rectally, nasally, buccally, vaginally, topically, parenterally, e.g., subcutaneously, intravenously, intramuscularly, intraperitoneally, intrathecally, intraventricularly, intrasternally and intracranially, by injection or infusion, or by means of an external suppository. Oral or intravenous administration is preferred.

[0154] The compounds of the present application can optionally also be used in combination with other active ingredients, each of which can be adjusted in amount and ratio by those skilled in the art depending on the particular condition and patient, and the clinical need, etc. The combination does not mean that the therapies or therapeutic agents must be administered and / or formulated for delivery together, although these methods of delivery are within the scope of the present application. The therapeutic agents used in combination can be administered simultaneously, before, or after one or more of the other additional therapies or therapeutic agents. The therapeutic agents or therapeutic regimens can be administered in any order.

[0155] "Optional" or "optionally" means that the subsequently described event or circumstance can or can not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not.

[0156] The application is further described in detail by reference to specific examples, which are not to be construed as limiting the scope of the application as claimed.

[0157] Examples

[0158] Example 1: Preparation of 3-(4-(6,7-dihydropyrazolo[l,5-a]pyrazin-5(4H)-yl)-6,7- dimethyl-9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0159] Step 1: Synthesis of compound A-2

[0160] Dissolve H2SO4(1.46 g, 14.92 mmol, 1.25 mL) in H2O (4 mL), keep the temperature at 25 °C, add A-1 (1 g, 4.97 mmol) at 25 °C with stirring, add NaNO2solution (428.97 mg, 6.22 mmol, 197.86 μL) dissolved in H2O (4 mL) dropwise with vigorous stirring at 0-5 °C in an ice bath, remove the ice bath, the system slowly warms up to 11 °C, stir for 1 hour; TLC monitoring, the reaction is complete, add sodium hydroxide solution dropwise at 20 °C to quench, then add saturated sodium bicarbonate solution to adjust pH = 7, filter, and oven dry to obtain A-2 (600 mg, 59% yield) as a white solid.

[0161] Step 2: Synthesis of compound A-3

[0162] Dissolve A-2 (600 mg, 2.97 mmol) in toluene (5 mL), 90 °C, N2atmosphere, stirring, POBr3(2.97 mmol) dissolved in toluene (5 mL) was added dropwise into the reaction system, the system changed from clear to turbid, 90 °C, N2atmosphere stirring overnight, TLC monitoring reaction, the reaction was completed, cooled to room temperature, the reaction liquid was poured into water, extracted with EA, the organic phase was dried over anhydrous sodium sulfate, rotary evaporation, column chromatography separation and purification (PE:DCM = 4:1) to obtain A-3 (400 mg, 50.8% yield) as a white solid.

[0163] Step 3: Synthesis of compound A-4

[0164] Dissolve A-3 (400 mg, 1.51 mmol) in DME (6 mL), add Cs2CO3(1.23 g, 3.77 mmol), Pd2(dba)3(1.38 g, 1.51 mmol), Xantphos (87.36 mg, 150.97 μmol), 2,6-dimethyl-3-methoxyaniline (239.70 mg, 1.59 mmol) in a 40 ml sample bottle, N2, 90 °C stirring overnight, TLC monitoring reaction, the reaction was completed, cooled to room temperature, the reaction liquid was poured into water, extracted with EA, the organic phase was dried over anhydrous sodium sulfate, rotary evaporation, column chromatography separation and purification (PE:DCM = 4:1) to obtain A-4 (320 mg, 63.2% yield) as a white solid.

[0165] Step 4: Synthesis of compound A-5

[0166] Dissolve malononitrile (132.42 mg, 2.00 mmol, 126.23 μL) in DME (4 mL), add sodium tert-butoxide (186 mg, 1.91 mmol), N2, room temperature stirring to clear, then add A-4 (320 mg, 954.55 μmol) and Pd(dppf)Cl2(60 mg, 95.45 μmol) to it, warmed to 90 °C reaction overnight, TLC monitoring, the reaction was completed, poured into water, extracted with EA, saturated brine, the organic phase was dried over anhydrous sodium sulfate, rotary evaporation, column chromatography separation and purification (PE:DCM = 1:1) to obtain A-5 (220 mg, 686.67 μmol, 71.94% yield) as a white solid.

[0167] Step 5: Synthesis of compound A-6

[0168] Dissolve A-5 (220 mg, 686.67 μmol) in concentrated sulfuric acid (10 mL), stir at room temperature for 1 hr, monitor the reaction by TLC, when the reaction is complete, add the system to ice water, extract with EA, wash with brine, dry the organic phase over anhydrous sodium sulfate, spin dry to get A-6 (170 mg, 73.1% yield).

[0169] Step 6: Synthesis of compound A-7

[0170] Dissolve A-6 (70 mg, 206.85 μmol) in super dry DMF (1 mL), add DMF-DMA (71.48 mg, 599.88 μmol), N2, microwave at 150 °C for 1 hr, monitor the reaction by TLC, MS shows the product is formed when the starting material is consumed. Purify by reverse phase column chromatography (water (0.1% NH4OH): ACN = 3:1 to elute the product, freeze dry to get A-7 (37 mg, 51.3% yield).

[0171] Step 7: Synthesis of compound A-8

[0172] Dissolve A-7 (56 mg, 160.74 μmol) in a microwave tube, add POCl3 (24.65 mg, 160.74 μmol, 14.98 μL), microwave at 150 °C for 10 min, monitor by TLC, when the reaction is complete, remove POCl3 by distillation under reduced pressure, dilute with water, add saturated sodium bicarbonate solution dropwise, extract with EA, spin dry the organic phase, purify by column chromatography (100% DCM) to get A-8 (50 mg, 84.8% yield).

[0173] Step 8: Synthesis of compound A-9

[0174] Dissolve A-8 (40 mg, 109 μmol) in a microwave tube, add tetrahydro pyrazolo[l,5-a]pyrazine (27 mg, 220 μmol), DIPEA (141 mg, 1.1 mmol) and dioxane (2 mL), microwave at 140 °C for 3 h, TLC-MS shows the starting material is consumed and the product is formed, concentrate under reduced pressure, purify by column chromatography (DCM:MeOH = 20: 1) to get the product as yellow oil (33 mg, 67%).

[0175] Step 9: Synthesis of Example 1 compound

[0176] Dissolve A-9 (33 mg, 73 μmol) in DCM (1 mL), add BBr3 (290 μmol) dropwise in an ice water bath, then react at room temperature for 40 min, monitor by LCMS, when the reaction is complete, quench with methanol, concentrate, purify by column chromatography (DCM:MeOH = 9: 1) to get the target molecule (25 mg yield 78%) as white solid.1 H NMR (400 MHz, Methanol-d4) δ 8.47 (s, 1H), 8.21 (s, 1H), 7.56 (d, J = 2.0 Hz, 1H), 7.12 (d, J = 8.3 Hz, 1H), 6.96 (d, J = 8.3 Hz, 1H), 6.28 (d, J = 2.0 Hz, 1H), 5.27 (s, 2H), 4.51 (dd, J = 5.9, 4.0 Hz, 2H), 4.49 - 4.45 (m, 2H), 2.57 (s, 3H), 2.54 (s, 3H), 1.74 (s, 3H), 1.69 (s, 3H); Mass: C 25 H 25 N7O [M+H] + Calculated: 440.2, Found: 440.2

[0177] Example 2: Preparation of 9-(3-hydroxy-2,6-dimethylphenyl)-6,7-dimethyl-3,9- dihydro-4H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-4-one

[0178] The compound of Example 2 (6 mg, yield: 41%, 99% purity) was prepared according to the demethylation procedure of Reference Example 1, as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 9.59 (s, 1H), 8.15 (s, 1H), 8.08 (s, 1H), 7.03 (d, J = 8.3 Hz, 1H), 6.94 (d, J = 8.3 Hz, 1H), 2.42 (s, 3H), 2.39 (s, 3H), 1.68 (s, 3H), 1.57 (s, 3H); Mass: C 19 H 18 N4O2; [M+H] + Calculated: 335.2, Found: 335.2.

[0179] Example 3: Preparation of 4-amino-9-(3-hydroxy-2,6-dimethylphenyl)-6,7-dimethyl- 1,9-dihydro-2H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-2-one

[0180] Step 1: Synthesis of Compound A-10

[0181] Dissolve A-5 (40 mg, 124.85 pmol) in super dry DCM (1 mL), stir at room temperature, dropwise add chlorosulfonyl isocyanate (194.37 mg, 1.37 mmol, 119.54 pL) in super dry DCM (1 mL), continue to react at room temperature for 3 h, then react at 45 °C overnight, TLC monitoring, the raw material is reacted completely, add ice water (7.97 mL), stand overnight, filter, and oven dry the filter cake to obtain A-10 (42 mg, 92% yield) as a light yellow solid.

[0182] Step 2: Synthesis of the compound of Example 3

[0183] The target molecule was prepared according to the demethylation method of Reference Example 1 (5 mg, 18% yield, 98% purity), white solid. 1 H NMR (400 MHz, Methanol-d4) d 8.12 (s, 1H), 6.96 (d, J = 8.3 Hz, 1H), 6.80 (d, J = 8.3 Hz, 1H), 2.35 (s, 3H), 2.33 (s, 3H), 1.73 (s, 3H), 1.67 (s, 3H); Mass spectrum: C 19 H 19 N5O2; [M+H] + Calculated: 350.2, Found: 350.2.

[0184] Example 4: Preparation of 3-(6,7-dimethyl-4-(methylamino)-9H-pyrido[3',2':4,5]pyrrolo[2,3- d]pyrimidin-9-yl)-2,4-dimethylphenol

[0185] Step 1: Synthesis of compound A-11

[0186] Weigh A-8 (20 mg, 54.52 pmol) and dissolve it in EtOH (1 mL), add DIPEA (28.49 pL, 163.56 pmol), methylammonium hydrochloride (3.68 mg, 54.52 pmol), reflux and react overnight, TLC monitoring, the reaction is complete, and the obtained crude product (30 mg) is directly used in the next step.

[0187] Step 2: Synthesis of the compound of Example 4

[0188] The target molecule was prepared according to the demethylation method of Reference Example 1 (5 mg, 18% yield, 98% purity), white solid. 1H NMR (400 MHz, Methanol-d4) δ 8.47 (s, 1H), 8.30 (s, 1H), 7.10 (d, J = 8.3 Hz, 1H), 6.94 (d, J = 8.3 Hz, 1H), 3.24 (s, 3H), 2.54 (s, 3H), 2.51 (s, 3H), 1.76 (s, 3H), 1.70 (s, 3H); Mass spectrum: C 20 H 21 N5O; [M+H] + Calculated: 348.2, Found: 348.2.

[0189] Example 5: Preparation of 3-(6,7-dimethyl-4-(propylamino)-9H-pyrido[3',2':4,5]pyrrolo[2,3- d]pyrimidin-9-yl)-2,4-dimethylphenol

[0190] The compound of Example 5 (10 mg, 53%, 99% purity) was prepared by the method of Reference Example 4 as a white solid. 1 H NMR (400 MHz, Methanol-d4) δ 8.47 (s, 1H), 8.30 (s, 1H), 7.10 (d, J = 8.3 Hz, 1H), 6.94 (d, J = 8.3 Hz, 1H), 3.24 (s, 3H), 2.54 (s, 3H), 2.51 (s, 3H), 1.76 (s, 3H), 1.70 (s, 3H); Mass spectrum: C 22 H 25 N5O; [M+H] + Calculated: 376.2, Found: 376.2.

[0191] Example 6: Preparation of 3-(4-(hydroxyamino)-6,7-dimethyl-9H-pyrido[3',2':4,5]pyrrolo[2,3- d]pyrimidin-9-yl)-2,4-dimethylphenol

[0192] The compound of Example 6 (5.5 mg, 18%, 96% purity) was prepared by the method of Reference Example 4 as a yellow solid. 1 H NMR (600 MHz, Methanol-d4) δ 8.44 (s, 1H), 8.28 (s, 1H), 8.16 (s, 1H), 7.71 (s, 1H), 7.09 (d, J = 8.2 Hz, 1H), 6.93 (d, J = 8.2 Hz, 1H), 2.52 (s, 3H), 2.46 (s, 3H), 1.77 (s, 3H), 1.72 (s, 3H); Mass spectrum: C 19 H19 N5O2; [M+H] + Calculated: 350.2, Found: 350.2.

[0193] Example 7: Preparation of 3-(4-hydrazinyl-6,7-dimethyl-9H-pyrido[3',2':4,5]pyrrolo[2,3- d]pyrimidin-9-yl)-2,4-dimethylphenol

[0194] The compound of Example 7 (9 mg, 28%, 90% purity) was prepared by the method of Reference Example 4 as a white solid.1H NMR (400 MHz, Methanol-d4) δ 8.52 (s, 1H), 8.36 (s, 1H), 7.11 (d, J = 8.3 Hz, 1H), 6.95 (d, J = 8.3 Hz, 1H), 4.62 (s, 2H), 2.54 (s, 3H), 2.50 (s, 3H), 1.75 (s, 3H), 1.70 (s, 3H); Mass spectrum: C 19 H 20 N6O; [M+H] + Calculated: 349.2, Found: 349.2.

[0195] Example 8: Preparation of N-(9-(3-hydroxy-2,6-dimethylphenyl)-6,7-dimethyl-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-4-yl)acetamide

[0196] Step 1: Synthesis of compound A-12

[0197] A-8 (20 mg, 54 μmol) was weighed into a microwave tube, NH4OH (450 mg, 12.5 mmol, 0.5 mL) and dioxane (1 mL) were added, and the mixture was subjected to microwave reaction at 150 °C for 40 min. After concentration under reduced pressure, the residue was dissolved in THF (1 mL), TEA (8.74 mg, 86.35 μmol, 12.04 μL) was added, and Ac2O (29.39 mg, 287.84 μmol, 27.16 μL) was added dropwise at 0 °C, followed by heating to reflux for 2 h. After the starting material was consumed, the mixture was dried, and column chromatography (DCM:MeOH = 9:1) was performed to give 20 mg of acylated intermediate A-12 as a white solid.

[0198] Step 2: Synthesis of the compound of Example 8

[0199] The compound of Example 8 (3.0 mg, 14%, 92% purity) was prepared by the demethylation method of Reference Example 1 as a white solid. 1H NMR (400 MHz, DMSO-d6) δ 11.05 (s, 1H), 9.51 (s, 1H), 8.69 (s, 1H), 7.79 (s, 1H), 7.08 (d, J = 8.3 Hz, 1H), 6.94 (d, J = 8.1 Hz, 1H), 2.46 (s, 3H), 2.41 (s, 3H), 2.35 (s, 3H), 1.67 (s, 3H), 1.56 (s, 3H); Mass: C 21 H 21 N5O2; [M+H] + Calculated: 376.2, Found: 376.2.

[0200] Example 9: Preparation of N-(9-(3-hydroxy-2,6-dimethylphenyl)-6,7-dimethyl-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-4-yl)propanamide

[0201] The compound of Example 9 (7 mg, 29%, 97% purity) was prepared according to the procedure of Reference Example 8 as a white solid. 1 H NMR (600 MHz, Methanol-d4) δ 8.44 (s, 1H), 8.21 (s, 1H), 7.32 (d, J = 8.4 Hz, 1H), 7.21 (d, J = 8.3 Hz, 1H), 2.65 (q, J = 7.5 Hz, 2H), 2.52 (s, 3H), 2.48 (s, 3H), 1.86 (s, 3H), 1.67 (s, 3H), 1.24 (t, J = 7.5 Hz, 3H); Mass: C 22 H 23 N5O2; [M+H] + Calculated: 390.2, Found: 390.2.

[0202] Example 10: Preparation of N-(9-(3-hydroxy-2,6-dimethylphenyl)-6,7-dimethyl-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-4-yl)formamide

[0203] Formic acid (27.61 mg, 599.90 μmol, 22.63 μL) and acetic anhydride (61.24 mg, 599.90 μmol, 56.60 μL) were mixed at 0 °C and stirred at room temperature for 45 min, then added dropwise to a solution of the compound of Example 1 (10 mg, 30.00 μmol) in THF (1 mL) at 0 °C, then warmed to 66 °C and stirred for 2 h. After spinning down, 1 ml of (MeOH-THF) was dissolved, and preparative reverse-phase liquid chromatography (water (0.1% formic acid):ACN = 2:1) gave the compound of Example 10 (2 mg, 18%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 11.10 (s, 1H), 9.65 (s, 1H), 9.52 (s, 1H), 8.83 (s, 1H), 8.59 (s, 1H), 7.07 (d, J = 8.3 Hz, 1H), 6.94 (d, J = 8.3 Hz, 1H), 2.46 (s, 3H), 2.42 (s, 3H), 1.66 (s, 3H), 1.56 (s, 3H); Mass spectrum: C 20 H 19 N5O2; [M+H] + Calculated: 362.2, Found: 362.2.

[0204] Example 11: Preparation of 3-(4-amino-2-chloro-6,7-dimethyl-9H-pyrrolo[3',2':4,5]pyrrolo[2,3- d]pyrimidin-9-yl)-2,4-dimethylphenol

[0205] Step 1: Synthesis of compound A-13

[0206] A-6 (200 mg, 591.01 μmol) and urea (10.00 g, 166.51 mmol) were weighed and heated to 150 °C for 72 hr, TLC-MS monitoring showed that the starting material was consumed, water was added and filtered under ultrasonic, the filtrate was concentrated and purified by column chromatography (DCM:MeOH = 20:1) to give the intermediate A-13 (125 mg, 58%) as a yellow solid.

[0207] Step 2: Synthesis of compound A-14

[0208] A-13 (125 mg, 343.03 μmol) was dissolved in POCl3(52.60 mg, 343.03 μmol, 31.97 μL) and reacted at 150 °C for 10 min under microwave, TLC-MS monitoring showed that the starting material was consumed, and the product was purified by column chromatography to give the intermediate A-14 (60 mg, 43% yield).

[0209] Step 3: Synthesis of compound A-15

[0210] A-14 (60 mg, 149.52 μmol) was weighed into 1,4-dioxane (1 mL), ammonia (4.50 g, 5 mL) was added, and the mixture was reacted at 120 °C for 30 min under microwave irradiation. The solvent was removed under reduced pressure to give 50 mg of crude product A-15, which was used directly in the next step.

[0211] Step 4: Synthesis of the compound of Example 11

[0212] The compound of Example 11 was prepared according to the demethylation method of Reference Example 1 (4 mg, 26%, 97% purity). 1 H NMR (400 MHz, Methanol-d4) δ 8.43 (s, 1H), 7.10 (d, J = 8.3 Hz, 1H), 6.94 (d, J = 8.3 Hz, 1H), 2.53 (s, 3H), 2.49 (s, 3H), 1.77 (s, 3H), 1.71 (s, 3H); Mass spectrum: C 19 H 18 ClN5O; [M+H] + Calculated: 368.1, Found: 368.1.

[0213] Example 12: Preparation of 3-(2,4-diamino-6,7-dimethyl-9H-pyrido[3',2':4,5]pyrrolo[2,3- d]pyrimidin-9-yl)-2,4-dimethylphenol

[0214] The compound of Example 12 was prepared according to the method of Reference Example 11 (with slight modification in the first step condition: microwave at 150 °C for 80 min) (10 mg, 72% yield, 99% purity) 1 H NMR (400 MHz, DMSO-d6) δ 9.35 (s, 1H), 8.28 (s, 1H), 8.14 (s, 1H), 6.99 (d, J = 8.2 Hz, 1H), 6.92 (s, 2H), 6.85 (d, J = 8.2 Hz, 1H), 6.25 (s, 2H), 2.34 (s, 3H), 2.33 (s, 3H), 1.71 (s, 3H), 1.61 (s, 3H); Mass spectrum: C 19 H 20 N6O; [M+H] + Calculated: 349.2, Found: 349.2.

[0215] Example 13: Preparation of 9-(3-hydroxy-2,6-dimethylphenyl)-6,7-dimethyl-1,9-dihydro-2H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidine-2,4(3H)-dione

[0216] The compound of Example 13 (6 mg, 60%, 97% purity) was prepared using intermediate A-13 as starting material, following the demethylation procedure of Example 1, as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 11.89 (s, 1H), 10.93 (s, 1H), 9.51 (s, 1H), 7.94 (s, 1H), 7.05 (d, J = 8.3 Hz, 1H), 6.92 (d, J = 8.2 Hz, 1H), 2.34 (s, 3H), 2.34 (s, 3H), 1.74 (s, 3H), 1.65 (s, 3H); Mass spectrum: C 19 H 18 N4O3; [M+H] + Calculated: 351.1, Found: 351.1.

[0217] Example 14: Preparation of 3-(4-amino-6,7-dimethyl-2-(propylamino)-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0218] Step 1: Synthesis of compound A-17

[0219] A-15 (20 mg, 52.38 μmol) was weighed and dissolved in 1,4-dioxane (0.5 mL), n-propylamine (359.50 mg, 6.08 mmol), DIPEA (37.10 mg, 287.06 μmol) was added and the reaction was carried out at 150 °C for 1 hr under microwave, TLC-MS monitoring showed the starting material was consumed, dried to get the crude product A-17 (22 mg) which was used directly for the next step.

[0220] Step 2: Synthesis of compound of Example 14

[0221] The compound of Example 14 (6 mg, 28%, 99% purity) was prepared using the demethylation procedure of Example 1, as a white solid. 1 H NMR (400 MHz, Methanol-d4) δ 8.35 (s, 1H), 7.09 (d, J = 8.3 Hz, 1H), 6.94 (d, J = 8.2 Hz, 1H), 3.29 (t, J = 7.0 Hz, 2H), 2.50 (s, 3H), 2.47 (s, 3H), 1.83 (s, 3H), 1.77 (s, 3H), 1.59 (h, J = 7.3 Hz, 2H), 0.91 (t, J = 7.4 Hz, 3H); Mass spectrum: C 19 H 18 N4O3; [M+H]+ Calculated: 391.2, Found: 391.2.

[0222] Example 15: Preparation of 3-(4-amino-6,7-dimethyl-2-(methylamino)-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0223] The compound of Example 15 (6 mg, 28%, 98% purity) was prepared according to the procedure of Reference Example 14 as a white solid. 1 H NMR (400 MHz, Methanol-d4) δ 8.35 (s, 1H), 7.09 (d, J = 8.3 Hz, 1H), 6.93 (d, J = 8.3 Hz, 1H), 2.90 (s, 3H), 2.50 (s, 3H), 2.47 (s, 3H), 1.83 (s, 3H), 1.77 (s, 3H); Mass spectrum: C 19 H 18 N4O3; [M+H] + Calculated: 363.1, Found: 363.1.

[0224] Example 16: Preparation of 3-(4-amino-2,6,7-trimethyl-9H-pyrido[3',2':4,5]pyrrolo[2,3- d]pyrimidin-9-yl)-2,4-dimethylphenol

[0225] Step 1: Synthesis of compound A-18

[0226] A-6 (60 mg, 177.3 μmol) was weighed into DMAc (1 mL), and then trimethyl orthoacetate (53 mg, 443 μmol) was added. The mixture was subjected to microwave reaction at 160 °C for 2 h. Intermediate A-18 (20 mg, 31%) was prepared by reverse phase liquid chromatography (water (0.1% formic acid): ACN = 3:2) as a white solid.

[0227] Step 2: Synthesis of compound A-19

[0228] A-18 (20 mg, 55.18 μmol) and POCl3 (8.46 mg, 55.18 μmol) were subjected to microwave reaction at 150 °C for 10 min. TLC-MS monitoring showed that the starting material was consumed. The mixture was dried and purified by column chromatography (PE:DCM = 1:1) to give intermediate A-19 (25 mg) as a white solid.

[0229] Step 3: Synthesis of compound A-20

[0230] A-19 (25 mg, 65.29 μmol) was dissolved in 1,4-dioxane (2 mL), ammonia (900.00 mg, 25.68 mmol) was added, and the mixture was reacted at 120 °C for 6 hr under microwave irradiation. The reaction mixture was concentrated to dryness to give crude A-20, which was used directly in the next step.

[0231] Step 4: Synthesis of the compound of Example 16

[0232] The compound of Example 16 (2 mg, 92% purity) was prepared from A-20 according to the demethylation method of Example 1 as a white solid. 1 HNMR (400 MHz, Methanol-d4) δ 8.71 (s, 1H), 7.13 (d, J = 8.3 Hz, 1H), 6.98 (d, J = 8.3 Hz, 1H), 2.66 (s, 3H), 2.60 (s, 3H), 2.55 (s, 3H), 1.78 (s, 3H), 1.72 (s, 3H); Mass: C 20 H 21 N5O; [M+H] + Calcd: 348.2, Found: 348.2.

[0233] Example 17: Preparation of 3-(4-amino-6-bromo-9H-pyrido[3',2':4,5]pyrrolo[2,3- d]pyrimidin-9-yl)-2,4-dimethylphenol

[0234] Step 1: Synthesis of compound B-2

[0235] B-1 (6.0 g, 21.28 mmol) was dissolved in NMP (50 mL), 2,6-dimethylpyridine (2.43 g, 22.70 mmol, 2.64 mL), 2,6-dimethyl-3-methoxyaniline (3.22 g, 21.28 mmol) were added by stirring, and the mixture was reacted at 130 °C overnight. After the reaction was completed, the system was added to water, stirred at room temperature for 20 minutes, filtered, and the filter cake was washed with water. The filter cake was dissolved in DCM and extracted. The organic phase was dried over anhydrous sodium sulfate, concentrated to dryness, and purified by silica gel column chromatography (PE:DCM = 3:1) to give intermediate product B-2 (3.6 g, 72%).

[0236] Step 2: Synthesis of compound B-3

[0237] Dissolve propiolonitrile (675.26 mg, 10.22 mmol) in DME (20.07 mL), protect under N2, add NaH (1.23 g, 30.67 mmol, 60% purity), stir at room temperature for 10 min to peach turbid liquid, add B-2 (3.6 g, 10.22 mmol), Pd(dppf)Cl2(373.97 mg, 511.10 μmol), the system turns into wine red, react at 110 °C for 2 h, monitor by LCMS, the reaction is complete. Filter the diatomite layer, elute with ethanol, and purify the filtrate by silica gel column chromatography (PE:DCM = 1:4) to obtain B-3 (2.6 g, 75.40% yield).

[0238] Step 3: Synthesis of compound B-4

[0239] Dissolve B-3 (2.6 g, 7.71 mmol) in concentrated sulfuric acid (60 mL), stir at rt for 40 min, monitor by TLC, the reaction is complete, dilute in ice water, extract with EA, wash with saturated brine, dry the organic phase with anhydrous sodium sulfate, and spin dry to obtain B-4 (2.7 g, 98% yield) as a yellow solid.

[0240] Step 4: Synthesis of compound B-5

[0241] Dissolve B-4 (2.6 g, 7.32 mmol) in triethyl orthoformate (50 mL), protect under N2 atmosphere, react at 200 °C for 30 min under microwave, monitor by TLC, the reaction is complete and the starting material is completely consumed, purify by column chromatography (DCM:MeOH = 20:1) after concentration to obtain B-5 (2.3 g, 86% yield).

[0242] Step 5: Synthesis of compound B-6

[0243] Dissolve B-5 (2.3 g, 6.30 mmol) in DCM (10 mL) and MeOH (10 mL), add Pd / C (10% on Carbon) (67.00 mg, 629.55 μmol), stir at room temperature under H2 atmosphere for 3 hours, monitor the reaction by TLC, the reaction is complete, filter the diatomite layer, and spin dry to obtain the intermediate product B-6 (2.1 g, 99% yield).

[0244] Step 6: Synthesis of compound B-7

[0245] Dissolve B-6 (600 mg, 1.79 mmol) in DMF (1.86 mL) and ACN (1.86 mL), add tert- butyl nitrite (276.74 mg, 2.68 mmol, 319.20 μL), CuBr2(479.52 mg, 2.15 mmol), warm to 80 °C, stir for 20 min, monitor the reaction by TLC, complete the reaction, cool to room temperature, filter, purify the product by preparative reverse phase liquid chromatography (water (0.1% formic acid):ACN = 2:5) to give intermediate B-7 (200 mg, 28.00% yield).

[0246] Step 7: Synthesis of compound B-8

[0247] Dissolve B-7 (200 mg, 500.95 μmol) in POCl3(10.97 g, 71.52 mmol, 6.67 mL), microwave at 150 °C for 20 min, monitor the reaction by TLC, complete the reaction, remove POCl3by distillation under reduced pressure, dilute with EA, extract with saturated sodium bicarbonate solution, wash the organic phase with brine, dry, spin dry, dissolve the sample in DMSO (1 ml), purify by preparative reverse phase liquid chromatography (water (0.1% formic acid):ACN = 1:2) to give intermediate B-8 (140 mg, 66% yield).

[0248] Step 8: Synthesis of compound B-9

[0249] Dissolve B-8 (140 mg, 335.18 μmol) in 1,4-dioxane (2 mL), add NH4OH (900.00 mg, 25.68 mmol, 1 mL), microwave at 100 °C for 20 min, monitor by LCMS, complete the reaction of the starting material, extract with brine and EA, distill the EA phase under reduced pressure to give B-9 (100 mg, 74.9%).

[0250] Step 9: Synthesis of Example 17 compound (i.e. B-10)

[0251] Prepare Example 17 compound (25 mg, 80%) as a white solid according to the de- methylation method of Reference Example 1. 1 H NMR (400 MHz, Methanol-d4) δ 8.98 (d, J = 2.1 Hz, 1H), 8.41 (d, J = 2.1 Hz, 1H), 8.30 (s, 1H), 7.12 (d, J = 8.3 Hz, 1H), 6.95 (d, J = 8.3 Hz, 1H), 1.77 (s, 3H), 1.72 (s, 3H); Mass spectrum: C 17 H 14 N5BrO [M+H] + Calculated: 384.1, Found: 384.1.

[0252] Example 18: Synthesis of 3-(4-amino-6-(3,6-dihydro-2H-pyran-4-yl)-9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0253] Compound B-10 (0.15 mmol, 1.0 eq), dihydropyranboronic acid (ester) compound (0.2 mmol, 1.2 eq), Pd(dppf)Cl2(0.02 mmol, 0.01 eq), K2CO3(0.45 mmol, 3.0 eq) were weighed and added 1 mL 1,4-dioxane: water (v / v = 1:1) and placed at 100 °C for 1 h. After completion of the reaction, it was cooled to room temperature, filtered, and purified by column chromatography (DCM / methanol = 20:1-10:1) to get the crude product, which was concentrated and purified by preparative high-performance liquid chromatography to get the compound of Example 18 (5.4 mg, yield: 9.4%). 1 H NMR (400 MHz, Methanol-d4) δ 8.82 (d, J = 2.1 Hz, 1H), 8.43 (d, J = 2.0 Hz, 1H), 8.29 (s, 1H), 7.13 (d, J = 8.3 Hz, 1H), 6.95 (d, J = 8.3 Hz, 1H), 6.36-6.38 (m, 1H), 4.38 (q, J = 2.8 Hz, 2H), 4.02 (t, J = 5.4 Hz, 2H), 2.68-2.72 (m, 2H), 1.78 (s, 3H), 1.73 (s, 3H); Mass: C 22 H 21 N5O2[M+H] + Calculated: 388.2, Found: 388.2.

[0254] Example 19: Preparation of 3-(4-amino-6-(tetrahydro-2H-pyran-4-yl)-9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0255] Example 18 compound (11 mg, 28.39 pmol) was dissolved in methanol (2 mL), Pd / C (302.15 pg, 2.84 pmol) was added, replaced with H2, and reacted at rt for 4 hr under a hydrogen atmosphere. The reaction was monitored by TLC, and after completion of the reaction, it was filtered, dried, dissolved in a small amount of methanol, and clarified. Purification by reverse-phase liquid chromatography (water (0.1% formic acid): ACN = 3:1) gave the compound of Example 19 (3.8 mg, yield: 31%, 90% purity) as a white solid. 1H NMR (400 MHz, Methanol-d4) δ 8.71 (d, J = 2.0 Hz, 1H), 8.27 (s, 1H), 8.26 (d, J = 2.0 Hz, 1H), 7.12 (d, J = 8.3 Hz, 1H), 6.95 (d, J = 8.3 Hz, 1H), 4.08 - 4.18 (m, 2H), 3.64 - 3.67 (m, 2H), 3.06 - 3.09 (m, 1H), 1.97 - 2.10 (m, 2H), 1.87 - 1.95 (m, 2H), 1.77 (s, 3H), 1.72 (s, 3H); Mass: C22H23N5O2 [M+H] + Calculated: 390.2, Found: 390.2.

[0256] Examples 20-41:

[0257] The corresponding compounds were prepared according to the synthetic procedure of Reference Example 17-19 using compound B-10 and the starting materials listed in Table 2 under similar reaction conditions as Example 18.

[0258] Table 2: Compounds of Examples 20-41

[0259] Example 42: Preparation of 3-(4-amino-6-(2-hydroxypropan-2-yl)-9H-pyrido[3',2':4,5]pyrrolo[2,3- d]pyrimidin-9-yl)-2,4-dimethylphenol

[0260] The compound of Example 40 (1.0 eq), meso-tetraphenylporphyrin cobalt (1.0 eq), tetraethylammonium borohydride (10 eq) were added into a reaction flask, then methanol and ethylene glycol dimethyl ether were added, oxygen was replaced for 3 times, and the reaction was stirred at 70 °C for 4 hours. TLC detection, after the reaction was completed, the pH was adjusted to 7 by adding aqueous sodium bicarbonate solution, EA extraction, the organic phase was washed with brine for 3 times, the organic phase was dried over anhydrous sodium sulfate, suction filtered, rotary evaporated, and separated and purified by reverse phase column chromatography (MeOH / H2O = 0.1-10) to obtain the compound of Example 42 (16.3 mg, yield: 40%); 1 H NMR (600 MHz, Methanol-d4) δ 9.03 (d, J = 2.1 Hz, 1H), 8.73 (d, J = 2.1 Hz, 1H), 8.49 (s, 1H), 7.12 (d, J = 8.3 Hz, 1H), 6.96 (d, J = 8.3 Hz, 1H), 1.76 (s, 3H), 1.71 (s, 6H), 1.70 (s, 3H); Mass: C20H21N5O2 [M+H] +Calculated: 364.2, Found: 364.2.

[0261] Example 43: Preparation of 3-(4-amino-6-morpholino-9H-pyrido[3',2':4,5]pyrrolo[2,3- d]pyrimidin-9-yl)-2,4-dimethylphenol

[0262] Step 1: Synthesis of compound B-11

[0263] B-8 (1 mmol, 1 eq.) was dissolved in DMF (5 mL), benzylamine (2 mmol, 2 eq.) and DIPEA (3 mmol, 3 eq.) were added, and the reaction was carried out at 80°C for 2 hours. LCMS monitoring showed that the raw material was completely reacted. Salt water and EA were added for extraction. The organic phase was washed and dried, and then distilled under reduced pressure. Column chromatography was used for separation and purification to obtain the intermediate product B-11.

[0264] Step 2: Synthesis of compound B-12

[0265] Compound B-11 (0.2 mmol, 1.0 eq), amine component morpholine (5 eq), tBuXPhos Pd G4 (20% eq.) and 2-tert-butyl-1,1,3,3-tetramethylguanidine (3.0 eq) were added to the reaction bottle, and then DMSO (2 mL) was added. Nitrogen was replaced for 3 times, and the reaction was stirred at 90°C for 2 hours. TLC detection showed that the reaction was completed. After filtration of the insoluble matter, water and EA were added for extraction. The organic phase was washed with salt water for 3 times, and then dried with anhydrous sodium sulfate. After suction filtration and rotary evaporation, reverse phase column chromatography (MeOH / H2O = 0.1-10) was used for separation and purification to obtain the product B-12.

[0266] Step 3: Synthesis of compound B-13

[0267] Compound B-12 (0.05 mmol, 1.0 eq) was dissolved in methanol (3 mL), and nitrogen was replaced for 3 times. Palladium carbon (0.1 eq.) was added, and nitrogen was replaced for 3 times. Hydrogen was replaced for 3 times, and the reaction was carried out at room temperature for 5 hours. TLC detection showed that the reaction was completed. After filtration of the palladium carbon, the solvent was removed by concentration. Reverse phase column chromatography (MeOH / H2O = 0.1-10) was used for separation and purification to obtain the product B-13.

[0268] Step 4: Synthesis of the compound of Example 43

[0269] Compound B-13 (0.03 mmol, 1.0 eq) was dissolved in dichloromethane (1 mL), and boron tribromide (3 eq) was added under ice water bath cooling. The reaction was carried out at room temperature for 3 hours. TLC detection showed that the reaction was completed. After the solvent was removed by concentration, the pH was adjusted to 7 by adding aqueous sodium bicarbonate solution, and the organic phase was extracted with ethyl acetate three times. The combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography, and then the compound of Example 43 was prepared by high performance liquid chromatography. 1 H NMR (400 MHz, chloroform-d) δ 8.51 (s, 1H), 8.30 (d, J = 2.4 Hz, 1H), 7.85 (d, J = 2.2 Hz, 1H), 7.07 (d, J = 8.3 Hz, 1H), 6.82 (d, J = 8.2 Hz, 1H), 6.27 (s, 2H), 3.94 - 4.01 (m, 4H), 3.30 (t, J = 4.8 Hz, 4H), 1.82 (s, 3H), 1.77 (s, 3H); Mass: C21H22N6O2 [M+H] + Calculated: 391.2, Found: 391.1.

[0270] Examples 44-48:

[0271] Referring to the synthesis method of Example 43, the amine component morpholine was replaced with the starting materials listed in Table 3 to prepare the corresponding compounds.

[0272] Table 3: Compounds of Examples 44-48

[0273] Example 49: Preparation of 3-(4-amino-7-(3,6-dihydro-2H-pyran-4-yl)-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0274] Step 1: Synthesis of compound C-2

[0275] C-1 (1 g, 4 mmol) was dissolved in concentrated sulfuric acid (10 mL), and NaNO2 (328.69 mg, 4.76 mmol) was added under stirring at 0 °C. The reaction was continued at 0 °C for 1 hr. LCMS monitoring showed that the starting material was consumed. The system was added dropwise into ice water, and a solid was precipitated. The solid was filtered and dried to obtain C-2 (900 mg, 89%) as a yellow-white solid.

[0276] Step 2: Synthesis of compound C-3

[0277] C-2 (900 mg, 3.56 mmol), Ag2C03 (2.06 g, 7.47 mmol) were added to toluene (10 mL), stirred at rt, benzyl bromide (421.81 mg, 3.91 mmol) was added, the mixture was yellow suspension, continued to react at rt for 5 hr, the system was gray suspension, TLC monitoring, the reaction was completed, filtered through celite, washed with ethanol, the filtrate was rotary evaporated to give C-3 (1.15 g, 94%) as an oil.

[0278] Step 3: Synthesis of compound C-4

[0279] C-3 (1.15 g, 3.35 mmol), potassium tert-butoxide (564.30 mg, 5.03 mmol), Pd2(dba)3 (153.50 mg, 167.63 μmol), Xantphos (193.99 mg, 335.26 μmol) and 2,6-dimethyl-3-methoxyaniline (537.35 mg, 3.55 mmol) were added to a reaction flask, replaced with N2, added dry toluene (10 mL), heated to 80 °C for 1 hr, cooled to rt. Directly mixed with silica gel, column chromatography separation and purification, eluted with 100% PE to give intermediate C-4 (850 mg, 61%) as a white oil.

[0280] Step 4: Synthesis of compound C-5

[0281] Malononitrile (7.26 mmol) was dissolved in DME (10 mL), NaH (60% dispersion in oil) (116.14 mg, 4.84 mmol) was added, stirred at rt for 30 min, C-4 (1 g, 2.42 mmol) and Pd(dppf)Cl2 (177.04 mg, 241.95 μmol) were added to the system, reacted at 100 °C overnight, TLC monitoring, most of the starting material was consumed, diluted with ethanol, mixed with silica gel, rotary evaporated, column chromatography separation and purification (PE:DCM = 1:3) to give C-5 (450 mg, 46%) as a white solid.

[0282] Step 5: Synthesis of compound C-6

[0283] C-5 (450 mg, 1.13 mmol) was dissolved in formamide (10 mL), stirred at 205 °C for 1 hr, TLC monitoring, the reaction was completed, cooled to rt, added to ice water, extracted with DCM, washed with brine, dried the organic phase with anhydrous sodium sulfate, concentrated to give C-6 crude product 450 mg, which was directly used in the next step.

[0284] Step 6: Synthesis of compound C-7

[0285] C-6 (450 mg, 1.06 mmol) was dissolved in MeOH (10 mL), Pd / C (45.02 mg, 423.05 μmol) was added, H2was replaced, stirred at room temperature overnight, the reaction was monitored, the raw material was completely consumed, filtered and rotary evaporated to get the intermediate product C-7 crude (310 mg, 87%).

[0286] Step 7: Synthesis of compound C-8

[0287] C-7 (300 mg, 894.56 μmol) and Cs2CO3(349.76 mg, 1.07 mmol) were dissolved in DMF (5 mL), stirred at rt, N-phenyl bis(trifluoromethanesulfonyl)imide (351.54 mg, 984.02 μmol) was added, the reaction was continued at rt for 1 h, TLC monitoring, the reaction was completed. Diluted with ice water, extracted with DCM, saturated brine was washed, the organic phase was dried with anhydrous sodium sulfate, concentrated to get C-8 crude product (330 mg, 79%).

[0288] Step 8: Synthesis of compound C-9

[0289] C-8 (330 mg, 706.00 μmol) was dissolved in DCM (5 mL), stirred at 0 °C, BBr3(4.24 mmol) was added, the reaction was continued at rt for 1 h, TLC monitoring, the reaction was completed, ice bath, methanol was added dropwise to quench, rotary evaporated, dissolved in a small amount of methanol and clarified, reversed phase liquid chromatography preparation (water (0.1% formic acid): ACN = 3:2) can get C-9 (250 mg, 78%) white solid.

[0290] Step 9: Synthesis of compound of example 49

[0291] Compound C-9 (0.15 mmol, 1.0 eq), the corresponding boronic acid (ester) compound 3,6-dihydro-4-boronic acid pinacol ester-2H-pyran (0.2 mmol, 1.2 eq), Pd(dppf)Cl2(0.015 mmol, 0.1 eq), K2CO3(0.45 mmol, 3.0 eq), 2 mL 1,4-dioxane: water (v / v = 1:1) was added and reacted at 100 °C for 1 h. After the reaction was completed, it was cooled to room temperature, filtered, separated and purified by column chromatography (DCM / methanol = 20:1-10:1) to get the crude product, concentrated, and prepared by high performance liquid chromatography to get the compound of example 49 (9.8 mg, yield: 57%); 1H NMR (600 MHz, Methanol-d4) δ 8.57 (d, J = 8.2 Hz, 1H), 8.22 (s, 1H), 7.55 (d, J = 8.2 Hz, 1H), 7.07 (d, J = 8.3 Hz, 1H), 6.90 (d, J = 8.3 Hz, 1H), 6.66-6.67 (m, 1H), 4.30 (q, J = 2.8 Hz, 2H), 3.88 (t, J = 5.5 Hz, 2H), 2.56-2.59 (m, 2H), 1.76 (s, 3H), 1.71 (s, 3H); Mass: C22H21N5O2 [M+H] + Calculated: 388.2, Found: 388.2.

[0292] Example 50: Preparation of 3-(4-amino-7-(tetrahydro-2H-pyran-4-yl)-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0293] The compound of Example 50 was prepared (11.9 mg, yield: 58%) from the compound of Example 49 by reference to the hydrogenation method of Example 19; 1 H NMR (600 MHz, Methanol-d4) δ 8.57 (d, J = 8.2 Hz, 1H), 8.22 (s, 1H), 7.55 (d, J = 8.2 Hz, 1H), 7.07 (d, J = 8.3 Hz, 1H), 6.90 (d, J = 8.3 Hz, 1H), 6.66-6.67 (m, 1H), 4.30 (q, J = 2.8 Hz, 2H), 3.88 (t, J = 5.5 Hz, 2H), 2.56-2.59 (m, 2H), 1.76 (s, 3H), 1.71 (s, 3H); Mass: C22H21N5O2 [M+H] + Calculated: 390.2, Found: 390.2.

[0294] Example 51: Preparation of 3-(4-amino-7-(pyrrolidin-1-yl)-9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0295] The compound of Example 51 was prepared (7.6 mg, yield: 44%) from the compound C-9 (20 mg, 1.0 eq) and the corresponding secondary amine compound tetrahydropyrrole (5.0 eq) by reference to the hydrogenation method of Example 19. 1H NMR (400 MHz, chloroform-d) δ 8.41 (s, 1H), 7.86 (d, J = 8.5 Hz, 1H), 7.01 (d, J = 8.2 Hz, 1H), 6.79 (d, J = 8.2 Hz, 1H), 6.39 (d, J = 8.5 Hz, 1H), 5.26 (s, 2H), 3.38-3.40 (m, 4H), 1.92-1.97 (m, 4H), 1.84 (s, 3H), 1.79 (s, 3H); Mass: C21H22N6O [M+H] + Calculated: 375.2, Found: 375.2.

[0296] Example 52: Preparation of 3-(4-amino-7-((2-methoxyethyl)amino)-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0297] Compound C-9 (20 mg, 1.0 eq), corresponding primary amine compound 2-methoxyethylamine (5.0 eq), DIPEA (0.3 mL), n-butanol (1 mL) were weighed and placed in a microwave reactor at 170 °C for 120 min. Upon completion of the reaction, it was cooled to room temperature, concentrated and purified by preparative HPLC to obtain the compound of Example 52 (9.0 mg, yield: 53%). 1 H NMR (400 MHz, chloroform-d) δ 8.41 (s, 1H), 7.86 (d, J = 8.5 Hz, 1H), 7.01 (d, J = 8.2 Hz, 1H), 6.79 (d, J = 8.2 Hz, 1H), 6.39 (d, J = 8.5 Hz, 1H), 5.26 (s, 2H), 3.38-3.40 (m, 4H), 1.92-1.97 (m, 4H), 1.84 (s, 3H), 1.79 (s, 3H); Mass: C21H22N6O [M+H] + Calculated: 379.2, Found: 379.2.

[0298] Example 53: Preparation of 3-(4-amino-7-(pyridin-3-ylethynyl)-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0299] Into a microwave reaction tube was placed C-9 (20 mg, 1.0 eq), the corresponding terminal alkyne compound 3-ethynylpyridine (1.5 eq), Pd(PPh3)4(0.1 eq), CuI (0.05 eq), triethylamine (8.0 eq), and anhydrous DMF (1 mL) as solvent, replaced with nitrogen, and warmed to 120 °C for 1 h. Upon completion of the reaction, EA was added for extraction, and the organic phase was concentrated. The compound of Example 53 (10 mg, yield: 55%) was obtained by preparative HPLC. 1 H NMR (400 MHz, Methanol-d4) δ 8.74 (d, J = 8.1 Hz, 1H), 8.70 (d, J = 2.1 Hz, 1H), 8.48 (dd, J = 5.1, 1.6 Hz, 1H), 8.34 (s, 1H), 8.01 (dt, J = 8.0, 1.9 Hz, 1H), 7.69 (d, J = 8.1 Hz, 1H), 7.43 (dd, J = 8.0, 5.0 Hz, 1H), 7.03 (d, J = 8.3 Hz, 1H), 6.88 (d, J = 8.3 Hz, 1H), 1.69 (s, 3H), 1.63 (s, 3H); Mass: C24H18N6O [M+H] + Calculated: 407.2, Found: 407.2.

[0300] Examples 54-78:

[0301] The synthesis method of Reference Example 49-53 was used to prepare the corresponding target compound using compound C-9 and the starting materials listed in Table 4.

[0302] Table 4: Compounds of Examples 54-78

[0303] Example 79: Preparation of 3-(4-amino-7-(2-hydroxypropan-2-yl)-9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0304] The compound of Example 63 (20 mg, 57.90 μmol) was dissolved in dilute sulfuric acid (2 mL), and a microwave reaction was performed at 90 °C for 1.5 h. The reaction was monitored by TLC-MS, and 70% of the starting material was consumed with the formation of the product. The pH was adjusted to weak alkalinity by adding saturated sodium bicarbonate solution, and EA was added for extraction. The organic phase was washed with saturated brine, dried, and concentrated. The sample was dissolved in an appropriate amount of methanol, and the compound of Example 79 (3.2 mg, yield: 15%) was obtained by preparative HPLC. 1HNMR (600 MHz, Methanol-d4) δ 8.61 (d, J = 8.1 Hz, 1H), 8.23 (s, 1H), 7.66 (d, J = 8.2 Hz, 1H), 7.09 (d, J = 8.3 Hz, 1H), 6.92 (d, J = 8.3 Hz, 1H), 1.76 (s, 3H), 1.70 (s, 3H), 1.50 (s, 6H); Mass: C20H21N5O2 [M+H] + Calculated: 364.2, Found: 364.2.

[0305] Example 80: Preparation of 3-(4-amino-7-(2-fluoroprop-2-yl)-9H-pyrido[3',2':4,5]pyrrolo[2,3- d]pyrimidin-9-yl)-2,4-dimethylphenol

[0306] The compound of Example 79 (9 mg, 24.77 μmol) was dissolved in DCM (1 mL) under N2 atmosphere, DAST (11.98 mg, 74.30 μmol) was added at -78 °C, the reaction was continued at -78 °C for 1 hr, LCMS monitoring, the reaction was completed, saturated sodium bicarbonate solution was added at low temperature, extracted with DCM, saturated brine was washed, the organic phase was dried over anhydrous sodium sulfate, rotary evaporation, sample was dissolved in ACN, high performance liquid chromatography preparation (0.1% formic acid) to obtain the compound of Example 80 (1 mg, yield: 10%), white solid. 1 H NMR (600 MHz, Methanol-d4) δ 8.55 (d, J = 8.1 Hz, 1H), 8.16 (s, 1H), 7.52 (dd, J = 8.2, 1.7 Hz, 1H), 6.98 (d, J = 8.2 Hz, 1H), 6.82 (d, J = 8.3 Hz, 1H), 1.66 (s, 3H), 1.60 (s, 3H), 1.54 (d, J = 2.8 Hz, 3H), 1.50 (d, J = 2.8 Hz, 3H); Mass: C20H20FN5O [M+H] + Calculated: 366.2, Found: 366.2.

[0307] Example 81: Preparation of 3-(4-amino-6-(3,6-dihydro-2H-pyran-4-yl)-9H-pyrazino[2',3':4,5]pyrrolo[2,3- d]pyrimidin-9-yl)-2,4-dimethylphenol

[0308] Step 1: Synthesis of compound D-2

[0309] In a reaction flask, D-1 (1.0 eq) and concentrated sulfuric acid were added, and the temperature was lowered to 0 °C with an ice water bath. Sodium nitrite (1.1 eq) was then slowly added, and the reaction was allowed to proceed for 1 h. TLC monitoring showed that the reaction was incomplete, so sodium nitrite (0.5 eq) was added, and the reaction was allowed to continue for 2 h. TLC monitoring showed that the reaction was complete. The reaction solution was slowly added dropwise to stirring ice water, and a solid precipitated. After the addition was complete, the solid was filtered, washed with pure water, and dried under vacuum to obtain a light yellow solid, D-2.

[0310] Step 2: Synthesis of compound D-3

[0311] In a reaction flask, D-2 (1.0 eq) and silver carbonate (2.0 eq) were added, and toluene was used as the solvent. Benzyl bromide (1.05 eq) was then added dropwise to the suspension, and the reaction was allowed to proceed at room temperature for 3 h. TLC monitoring showed that the reaction was complete. The reaction mixture was filtered through diatomite, and the filter cake was washed with ethyl acetate. The filtrate was concentrated, diluted with ethyl acetate, and washed with saturated sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the product D-3 as an oily liquid.

[0312] Step 3: Synthesis of compound D-4

[0313] In a reaction flask, D-3 (1.0 eq), 2,6-dimethyl-3-methoxybenzene (1.05 eq), a catalyst Pd2(dba)3 (0.05 eq), a ligand Xantphos (0.1 eq), and potassium tert-butoxide (2.0 eq) were added. Anhydrous toluene was used as the reaction solvent, and the reaction was allowed to proceed at 80 °C for 45 min under nitrogen. After the reaction was complete, the reaction solution was concentrated, and column chromatography was used for purification to obtain the product D-4 as an oily liquid.

[0314] Step 4: Synthesis of compound D-5

[0315] In a reaction flask, malononitrile (2.1 eq) and anhydrous ethylene glycol dimethyl ether were added, and NaH (60% dispersion in oil) (3.0 eq) was added under an ice water bath. The mixture was stirred for 30 min. Then, D-4 (1.0 eq) and 1,1-bis(diphenylphosphino)ferrocene palladium dichloride (0.1 eq) were added. The reaction was allowed to proceed at 110 °C for 12 h under nitrogen. TLC monitoring showed that the reaction was complete. The reaction mixture was filtered through diatomite, and the filter cake was washed with ethyl acetate. The filtrate was concentrated, and column chromatography was used for purification to obtain the intermediate product D-5.

[0316] Step 5: Synthesis of compound D-6

[0317] In a reaction flask, D-5 (1.0 eq) was added, and formamide was used as the reaction reagent and solvent. The reaction was allowed to proceed at 205 °C for 40 min. TLC-MS and TLC monitoring showed that the reaction was complete. The reaction was extracted with dichloromethane to obtain an organic layer, which was concentrated and dried to obtain the crude intermediate product D-6. The product was directly used in the next step.

[0318] Step 6: Synthesis of compound D-7

[0319] To a reaction flask was added D-6 (1.0 eq) and Pd / C (10% on Carbon (wetted with ca. 55% Water)) (0.15 eq) in methanol, replaced with hydrogen gas, stirred at 60 °C for 4 h under hydrogen atmosphere. Upon completion of the reaction, filtered through celite and washed with methanol. The filtrate was concentrated to give the intermediate D-7, which was used directly in the next step.

[0320] Step 7: Synthesis of compound D-8

[0321] To a reaction flask was added D-7 (1.0 eq) in THF, followed by the addition of N- phenylbis(trifluoromethanesulfonimide) (1.0 eq) and triethylamine (2.0 eq). The reaction mixture was stirred at room temperature for 1 h. Upon completion of the reaction, the reaction mixture was extracted with dichloromethane to give the organic layer, which was concentrated and dried. The residue was purified by column chromatography to give the intermediate D-8.

[0322] Step 8: Synthesis of compound D-9

[0323] To a reaction flask was added D-8 (1.0 eq) in dichloromethane, followed by the slow addition of boron tribromide (3.0 eq). The reaction mixture was stirred at room temperature for 1 h. Upon completion of the reaction, the reaction mixture was quenched with methanol in an ice-water bath. The excess solvent was removed by concentration. The residue was purified by reverse phase column chromatography to give the intermediate D-9.

[0324] Step 9: Synthesis of compound of Example 81

[0325] To a microwave reaction tube was added D-9 (1.0 eq), the corresponding boronic acid (ester) compound 3,6-dihydro-4-boronic acid pinacol ester-2H-pyran (1.2 eq), 1,1- bis(diphenylphosphino)ferrocene palladium dichloride (0.1 eq), potassium carbonate (2.0 eq), and dioxane and water (v:v = 2:1). The reaction tube was replaced with N2and heated to 110 °C for 1 h under microwave. Upon completion of the reaction, the reaction mixture was cooled to room temperature, filtered, and purified by column chromatography (DCM / methanol = 20:1-10:1) to give the crude product. The compound of Example 81 (4.3 mg, yield: 26%) was obtained by preparative high-performance liquid chromatography. 1 H NMR (400 MHz, Methanol-d4) δ 8.56 (s, 1H), 8.49 (s, 1H), 7.13 (d, J = 8.3 Hz, 1H), 6.98-6.99 (m, 2H), 4.42-4.43 (m, 2H), 4.00-4.02 m, 2H), 2.82-2.84 (m, 2H), 1.79 (s, 3H), 1.73 (s, 3H); Mass spectrum: calculated for C21H21N6O2[M+H]+: 389.2, found: 389.2.

[0326] Example 82: Preparation of 3-(4-amino-6-(tetrahydro-2H-pyran-4-yl)-9H- pyrazino[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0327] The compound of Example 82 was prepared from the compound of Example 81 by reference to the hydrogenation method of Example 19 (8.6 mg, yield: 68%); 1 HNMR (400 MHz, Methanol-d4) δ 8.56 (s, 1H), 8.49 (s, 1H), 7.13 (d, J = 8.3 Hz, 1H), 6.98 (d, J = 8.3 Hz, 1H), 4.13-4.17 (m, 2H), 3.65-3.71 (m, 2H), 3.32-3.37 (m, 1H), 1.98-2.23 (m, 4H), 1.80 (s, 3H), 1.73 (s, 3H); Mass spectrum: [M+H]+calcd for C21H23N6O2: 391.2, found: 391.2.

[0328] Example 83: Preparation of 3-(4-amino-6-(pyrrolidin-1-yl)-9H-pyrazino[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0329] A microwave reaction vial was charged with compound D-9 (20 mg, 1.0 eq), the corresponding secondary amine compound pyrrolidine (5.0 eq), tetrahydrofuran (1 mL), and placed in a microwave reactor at 100 °C for 80 min. Upon completion, the reaction was cooled to room temperature, concentrated, and purified by preparative HPLC to give the compound of Example 83 (3.4 mg, yield: 42%); 1 HNMR (400 MHz, Methanol-d4) δ 8.42 (s, 1H), 7.89 (s, 1H), 7.11 (d, J = 8.3 Hz, 1H), 6.96 (d, J = 8.3 Hz, 1H), 3.67 (q, J = 6.4, 5.1 Hz, 4H), 2.19–2.11 (m, 4H), 1.81 (s, 3H), 1.74 (s, 3H); Mass spectrum: [M+H]+calcd for C20H22N7O: 376.2, found: 376.2.

[0330] Example 84: Preparation of 3-(4-amino-6-(cyclopropylamino)-9H-pyrazino[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0331] Take compound D-9 (20 mg, 1.0 eq), the corresponding primary amine compound cyclopropylamine (5.0 eq), DIPEA (5.0 eq), add n-butanol (1 mL), and place in a microwave reaction at 170°C for 120 min. After the reaction is completed, cool to room temperature, concentrate, and prepare the compound of Example 84 (1.5 mg, yield: 9%) by high performance liquid chromatography. 1 H NMR (400 MHz, Methanol-d4) δ 8.42 (s, 1H), 8.04 (s, 1H), 7.12 (d, J = 8.4 Hz, 1H), 6.97 (d, J = 8.4 Hz, 1H), 2.79-2.82 (m, 1H), 1.81 (d, J = 2.3 Hz, 3H), 1.74 (s, 3H), 0.90-0.92 (m, 2H), 0.61-0.63 (m, 2H); Mass spectrum: [M+H]+calcd for C19H20N7O: 362.2, found: 362.2.

[0332] Example 85: Preparation of 3-(4-amino-6-(cyclopropylethynyl)-9H-pyrazino[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0333] A microwave reaction tube is added with D-9 (20 mg, 1.0 eq), the corresponding terminal alkyne compound cyclopropylacetylene (1.5 eq), Pd(PPh3)4 (0.1 eq), CuI (0.05 eq), triethylamine (8.0 eq), and then anhydrous DMF (1 mL) is added as a solvent, replaced with nitrogen, warmed to 120°C for 1 h, and after the reaction is completed, extracted with EA to obtain an organic phase, concentrated, and prepared by high performance liquid chromatography to obtain the compound of Example 85 (5.2 mg, yield: 32%); 1 H NMR (400 MHz, Methanol-d4) δ 8.42 (s, 1H), 8.04 (s, 1H), 7.12 (d, J = 8.4 Hz, 1H), 6.97 (d, J = 8.4 Hz, 1H), 2.79-2.82 (m, 1H), 1.81 (d, J = 2.3 Hz, 3H), 1.74 (s, 3H), 0.90-0.92 (m, 2H), 0.61-0.63 (m, 2H); Mass spectrum: [M+H]+calcd for C19H20N7O: 362.2, found: 362.2.

[0334] Examples 86-111:

[0335] Referring to the synthesis method of Examples 81-85, the corresponding target compounds are prepared using compound D-9 and the raw materials listed in Table 5.

[0336] Table 5: Compounds of Examples 86-111

[0337] Example 112: Preparation of 3-(4-amino-6-(2-hydroxypropan-2-yl)-9H-pyrazino[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0338] Using the compound of Example 92 as the starting material, the compound of Example 112 (2.4 mg, yield: 29%) was prepared by referring to the synthetic method of Example 79; 1 HNMR (400 MHz, methanol-d4) δ8.73 (s, 1H), 8.51 (s, 1H), 7.13 (d, J = 8.3 Hz, 1H), 6.98 (d, J = 8.3 Hz, 1H), 1.83-1.71 (m, 12H); mass spectrum: C19H21N6O2 [M+H] + calculated value: 365.2, found value: 365.2.

[0339] Example 113: Preparation of 3-(4-amino-6-(2-fluoropropan-2-yl)-9H-pyrazino[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0340] Using the compound of Example 112 as the starting material, the compound of Example 113 (6.3 mg, yield: 69%) was prepared by referring to the synthetic method of Example 80; 1 H NMR (400 MHz, methanol-d4) δ 8.68 (s, 1H), 8.50 (s, 1H), 7.11 (d, J = 8.3 Hz, 1H), 6.96 (d, J = 8.3 Hz, 1H), 1.92 (s, 3H), 1.87 (s, 3H), 1.78 (s, 3H), 1.72 (s, 3H); mass spectrum: C19H20FN6O [M+H] + calculated value: 367.2, found value: 367.2.

[0341] Example 114: Preparation of 3-(4-amino-6-cyclopropyloxy-9H-pyrazino[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0342] Step 1: Synthesis of compound D-10

[0343] Into a reaction vial was placed D-9 (1.0 eq), followed by imidazole (4.0 eq), DMF as solvent, followed by TBDMSC1 (1.5 eq), stirred at 40 °C overnight, reaction was complete, extracted with EA, obtained organic layer, concentrated, purified by column chromatography to get intermediate D-10.

[0344] Step two: synthesis of compound of example 114

[0345] Into a microwave reaction tube was placed D-10 (1.0 eq), corresponding cyclopropanol (8.0 eq), followed by THF (2 mL) as solvent, heated to 100 °C for 2 h, reaction was complete, extracted with EA, obtained organic layer, concentrated, purified by column chromatography (DCM / methanol = 20:1-10:1) to get crude product, concentrated, purified by preparative HPLC to get compound of example 114 (0.9 mg, yield: 37%); 1 H NMR (400 MHz, Methanol-d4) δ 8.39 (s, 1H), 8.10 (s, 1H), 7.10 (d, J = 8.4 Hz, 1H), 6.94 (d, J = 8.3 Hz, 1H), 6.15-6.25 (m, 1H), 2.02-2.27 (m, 4H), 1.77 (s, 3H), 1.71 (s, 3H); Mass: [M+H]+calcd for C19H19N6O2: 363.2, found: 363.2.

[0346] Examples 115-120:

[0347] The synthesis of reference example 114 was followed using compound D-10 and the starting materials listed in Table 6 to prepare the corresponding target compounds.

[0348] Table 6: Compounds of examples 115-120

[0349] Example 121: Preparation of 3-(4-amino-7-(thiophen-2-yl)-9H-pyrazino[2',3':4,5]pyrrolo[2,3- d]pyrimidin-9-yl)-2,4-dimethylphenol

[0350] Step 1: synthesis of compound E-2

[0351] Malononitrile (8.87 g, 134.2 mmol) was dissolved in DME (120 mL), and NaH (60% w / w, 5.58 g, 139.6 mmol) was slowly added. After reacting at room temperature for 30 min, compound E-1 (10 g, 67.1 mmol) was added. After reacting at room temperature for 1 h, the temperature was raised to 110°C and reacted for 2 h. After the reaction was complete, the mixture was cooled to room temperature. The DME was first spin-dried, then washed with 1 M HCl, and filtered to obtain a filter cake. The filter cake was then washed with water and collected to obtain 7.28 g of intermediate product E-2 with a yield of 61%.

[0352] Step 2: Synthesis of compound E-3

[0353] E-2 (4.2 g, 23.6 mmol) was dissolved in DMF (50 mL), and 2,6-dimethyl-3-methoxyaniline (10.7 g, 70.8 mmol) was added. The reaction was refluxed at 150°C for 24 h. After the reaction was complete, the mixture was cooled to room temperature, quenched with water, and extracted with EA. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and dried by spin drying. The intermediate product E-3 (1.3 g) was separated and purified by column chromatography (PE:EA, 0→100%). The yield was 19%.

[0354] Step 3: Synthesis of compound E-4

[0355] Compound E-3 (1.3 g, 4.43 mmol) was dissolved in DMF (26 mL), and NBS (0.95 g, 5.32 mmol) was added. The mixture was reacted at room temperature for 1 h. After the reaction was complete, an appropriate amount of water was added, stirred for 20 min, and filtered to obtain a filter cake. The filter cake was then washed with water and dried with a diaphragm pump to obtain 0.91 g of the intermediate product E-4 with a yield of 55%.

[0356] Step 4: Synthesis of compound E-5

[0357] Compound E-4 (2.92 g, 7.87 mmol) was dissolved in triethyl orthoformate (708.3 mmol, 118 mL), heated to 120°C, and reacted overnight. After all the raw materials were reacted to the intermediate, the mixture was cooled to room temperature, the triethyl orthoformate was spin-dried, and ammonia (7M in MeOH, 188 mL, 1.31 mol) was added. When the ratio of raw material to product reached 1:1, the reaction was stopped, the mixture was spin-dried, and preparative separation and purification were performed to obtain E-5 (1.27 g, yield 47%). 1H NMR (400 MHz, chloroform-d) δ 8.61 (s, 1H), 8.55 (s, 1H), 7.21-7.19 (d, J = 8.0 Hz, 1H), 6.98-6.95 (d, J = 12.0 Hz, 1H), 6.47 (s, 2H), 3.87 (s, 3H), 1.85 (s, 3H), 1.77 (s, 3H); Mass: C 17 H 15 BrN6O [M+H] + Calculated: 399.0, Found: 399.0.

[0358] Step 5: Synthesis of compound E-6

[0359] Dissolve E-5 (330 mg, 706.00 pmol) in DCM (5 mL), 0 °C, add BBr3 (4.24 mmol) by stirring, continue to react at room temperature for 1 hr, TLC monitoring, reaction is complete, ice bath, dropwise addition of methanol to quench, rotary evaporation, then dissolved in a small amount of methanol to clarify, reverse phase liquid chromatography preparation (water (0.1% formic acid): ACN = 3:2) to obtain the intermediate product E-6, white solid.

[0360] Step 6: Synthesis of compound of Example 121

[0361] Take compound E-6 (0.15 mmol, 1.0 eq), the corresponding boronic acid (ester) compound 2-thiophene boronic acid (0.2 mmol, 1.2 eq), Pd(dppf)Cl2(0.015 mmol, 0.1 eq), K2CO3(0.45 mmol, 3.0 eq), add 2 mL 1,4-dioxane: water (v / v = 1:1), 100 °C for 1 h. Reaction is complete, cool to room temperature, filter, column chromatography separation and purification (DCM / methanol = 20:1-10:1) to obtain the crude product, concentrate, high performance liquid chromatography preparation to obtain the compound of Example 121 (8.5 mg, yield: 53%); 1 H NMR (600 MHz, methanol-d4) δ 9.00-8.95 (m, 1H), 8.38 (d, J = 2.3 Hz, 1H), 7.75 (q, J = 3.5 Hz, 1H), 7.48-7.43 (m, 1H), 7.11-7.14 (m, 2H), 6.97 (d, J = 8.2 Hz, 1H), 1.86 (s, 3H), 1.81 (s, 3H); Mass: C20H16N6OS [M+H] + Calculated: 389.1, Found: 389.3.

[0362] Example 122: Preparation of 3-(4-amino-7-morpholino-9H-pyrazino[2',3':4,5]pyrrolo[2,3- d]pyrimidin-9-yl)-2,4-dimethylphenol

[0363] Compound E-6 (20 mg, 1.0 eq), corresponding secondary amine compound morpholine (5.0 eq) were weighed out, 1 mL of tetrahydrofuran was added, and the reaction was left to proceed overnight at 120 °C. Upon completion of the reaction, the reaction was cooled to room temperature, concentrated, and the compound of Example 122 (15.4 mg, yield: 76%) was obtained by preparative high-performance liquid chromatography; 1 H NMR (600 MHz, DMSO-d6) δ 9.40 (s, 1H), 8.21 (s, 1H), 8.15 (s, 1H), 6.97 (d, J = 8.3 Hz, 1H), 6.83 (d, J = 8.2 Hz, 1H), 3.65 - 3.60 (m, 4H), 3.39 (t, J = 4.9 Hz, 4H), 1.65 (s, 3H), 1.56 (s, 3H); Mass: C20H21N7O2 [M+H] + Calculated: 392.2, Found: 392.2.

[0364] Examples 123-136:

[0365] The corresponding target compounds were prepared according to the synthetic procedure of Reference Examples 121-122 using compound E-6 and the starting materials listed in Table 7.

[0366] Table 7: Compounds of Examples 123-136

[0367] Examples 137-142:

[0368] The corresponding target compounds were prepared according to the synthetic procedure of Reference Examples 114 and 121 using compound E-6 and the starting materials listed in Table 8.

[0369] Table 8: Compounds of Examples 137-142

[0370] Example 143: Preparation of 3-(4-amino-7-methyl-6-(thiophen-3-yl)-9H-pyrido[3',2':4,5]pyrrolo[2,3- d]pyrimidin-9-yl)-2,4-dimethylphenol

[0371] Step 1: Synthesis of compound F-2

[0372] 3-Bromo-2-chloro-6-methyl-5-nitropyridine F-1 (25.35 g, 100.8 mmol) was added to a sealed tube, followed by NMP (90 mL), 2,6-lutidine (21.6 g, 201.6 mmol), and 2,6-dimethyl-3-methoxyaniline (22.83 g, 151.2 mmol). The temperature was raised to 130°C and the reaction was allowed to react for 5 days. After the reaction was complete, the mixture was cooled to room temperature and the reaction solution was slowly poured into 0.5N HCl (1.25 L) while stirring. A gelatinous substance was formed at the bottom of the beaker. The mixture was filtered and the filter residue was rinsed with water. The filter residue was collected and dissolved in DCM, dried over anhydrous sodium sulfate, filtered, and dried. The product F-2 (24.79 g) was separated and purified by column chromatography (DCM:PE, 0→100%). The yield was 67%.

[0373] Step 2: Synthesis of Compound F-3

[0374] NaH (60% w / w, 9.02 g, 225.5 mmol) was weighed and added to DME (200 mL) for later use. Malononitrile (14.91 g, 225.5 mmol) was dissolved in DME (120 mL) and the NaH DME solution was slowly added dropwise. After the reaction for 1 h, compound F-2 (33 g, 90.2 mmol) and Pd(dppf)Cl2·DCM (3.68 g, 4.5 mmol) were added. Under N2 protection, the temperature was raised to 95°C and the reaction was refluxed for 2 h. After the reaction was complete, the mixture was cooled to room temperature and quenched with saturated aqueous ammonium chloride solution. The mixture was extracted with DCM, and the organic phase was washed with water and saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, and dried by spin drying. The product F-3 33.2 g was separated and purified by column chromatography (PE:EA, 0→100%). The yield was 96%.

[0375] Step 3: Synthesis of Compound F-4

[0376] Compound F-3 (13 g, 28.5 mmol) was dissolved in THF (130 mL), di-tert-butyl dicarbonate (27.97 g, 128.2 mmol) was added, and DMAP (3.48 g, 28.5 mmol) was slowly added. The temperature was raised to 60°C, and DMAP (5.22 g, 42.7 mmol) was slowly added. After the reaction was complete, the mixture was cooled to room temperature, quenched with water, and extracted with EA. The organic phase was washed with water and saturated brine, and the organic phase was dried over anhydrous sodium sulfate, filtered, and dried by spin drying. The product F-4 (10.54 g) was separated and purified by column chromatography (PE:EA, 0→100%). The yield was 67%.

[0377] Step 4: Synthesis of Compound F-5

[0378] Compound F-4 (7.83 g, 17.3 mmol) was dissolved in DCM:MeOH (1:1, 280 mL), Pd / C (2.95 g, 1.4 mmol, 5% w / w) was added, the reaction system was replaced with H2, and the reaction was carried out overnight under H2. After the reaction was completed, the palladium carbon was filtered with diatomite, the filtrate was collected, and rotary evaporation was performed. Purification was performed by column chromatography (PE:EA, 0→100%) to obtain the product F-5 7 g, with a yield of 95%.

[0379] Step 5: Synthesis of compound F-6

[0380] Compound F-5 (5 g, 11.9 mmol) was dissolved in super-dry MeCN (85 mL), CuBr2 (2.65 g, 11.9 mmol) was added, the air was replaced with N2, tert-butyl nitrite (1.46 g, 14.2 mmol) was added under N2 protection, the temperature was raised to 60°C, and the reaction was carried out for 1.5 h. After the reaction was completed, rotary evaporation was performed, and purification was performed by column chromatography (PE:EA, 0→100%) to obtain the product F-6 (3.92 g, yield 68%).

[0381] Step 6: Synthesis of compound F-7

[0382] Compound F-6 (3.52 g, 7.3 mmol) was weighed into a bottle, formamide (35.2 mL) was added, the temperature was raised to 205°C, and the reaction was carried out for 40 min. Water was added for quenching, extraction was performed with EA, the organic phase was washed with water and saturated brine, the organic phase was dried over anhydrous sodium sulfate, filtration was performed, rotary evaporation was performed, and purification was performed by column chromatography (PE:EA, 0→100%) to obtain the product F-7 (2 g, yield 67%). 1 H NMR (400 MHz, CD3OD) δ 8.50 (s, 1H), 8.24 (s, 1H), 7.20 (d, J = 8.0 Hz, 1H), 6.95-6.94 (d, J = 4.0 Hz 1H), 5.61 (s, 2H), 3.86 (s, 3H), 2.69 (s, 3H), 1.83 (s, 3H), 1.76 (s, 3H); Mass spectrum: C 19 H 18 BrN5O [M+H] + Calculated: 411.0, Found: 411.0.

[0383] Step 7: Synthesis of compound F-8

[0384] F-7 (1.0 eq) was added to a reaction bottle, dichloromethane was added as a solvent, and after cooling in an ice bath, boron tribromide (3.0 eq) was slowly added dropwise. Stirring was performed at room temperature for 1 h, TLC monitoring was performed, the reaction was completed, methanol was added for quenching under an ice water bath, excess solvent was removed by concentration, and reverse phase column chromatography purification was performed to obtain the intermediate product F-8.

[0385] Step 8: Synthesis of compound of Example 143

[0386] Compound F-8 (1.0 eq), corresponding boronic acid (ester) compound thiophene-3-boronic acid (1.2 eq), 1,1-bis(diphenylphosphino)ferrocene palladium dichloride (0.1 eq) and potassium carbonate (2.0 eq) were added to a reaction flask, followed by dioxane and water (v:v = 2:1), replaced with N2, and reacted at 110 °C for 1 h under N2protection. After the reaction was completed, it was cooled to room temperature, filtered, separated and purified by column chromatography (DCM / methanol = 20:1-10:1) to obtain the crude product, which was concentrated and prepared by high performance liquid chromatography to obtain the compound of Example 143 (19 mg, yield: 60%); 1 H NMR (400 MHz, Methanol-d4) δ 8.73 (s, 1H), 8.42 (s, 1H), 7.11 (d, J = 8.3 Hz, 1H), 6.95 (d, J = 8.2 Hz, 1H), 6.36 (d, J = 1.6 Hz, 1H), 5.84 (s, 1H), 2.57 (s, 3H), 1.77 (s, 3H), 1.71 (s, 3H); Mass: C21H18N5OF3 [M+H] + Calculated: 402.1, Found: 402.2.

[0387] Example 144: Preparation of 3-(4-amino-7-methyl-6-(3,3,3-trifluoroprop-1-en-2-yl)- 9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0388] The compound of Example 144 was prepared using the synthesis method of Example 143 (2.4 mg, yield: 68%); 1 H NMR (400 MHz, Methanol-d4) δ 8.73 (s, 1H), 8.42 (s, 1H), 7.11 (d, J = 8.3 Hz, 1H), 6.95 (d, J = 8.2 Hz, 1H), 6.36 (d, J = 1.6 Hz, 1H), 5.84 (s, 1H), 2.57 (s, 3H), 1.77 (s, 3H), 1.71 (s, 3H); Mass: C21H18N5OF3 [M+H] + Calculated: 414.2, Found: 414.2.

[0389] Example 145: Preparation of 3-(4-amino-7-methyl-6-(1,1,1-trifluoropropan-2-yl)-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol Example 145: Preparation of 3-(4-amino-7-methyl-6-(1,1,1-trifluoropropan-2-yl)-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0390] The compound of Example 145 was prepared from the compound of Example 144 according to the hydrogenation method of Example 19 (11.9 mg, yield: 78%); 1 HNMR (400 MHz, Methanol-d4) δ 8.96 (s, 1H), 8.44 (s, 1H), 7.10 (d, J = 8.4 Hz, 1H), 6.95 (d, J = 8.3 Hz, 1H), 4.11-4.19 (m, H), 2.68 (s, 3H), 1.79-1.66 (m, 9H); Mass: C21H20N5OF3 [M+H] + Calculated: 416.2, Found: 416.2.

[0391] Example 146: Preparation of 3-(4-amino-7-methyl-6-(prop-1-en-2-yl)-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0392] The compound of Example 146 was prepared according to the synthetic method of Example 143 (32 mg, yield: 33%); 1 HNMR (600 MHz, Methanol-d4) δ 8.45 (s, 1H), 8.23 (s, 1H), 7.09 (d, J = 8.3 Hz, 1H), 6.93 (d, J = 8.2 Hz, 1H), 5.37 (t, J = 1.7 Hz, 1H), 5.01 (t, J = 1.7 Hz, 1H), 2.55 (s, 3H), 2.17 (s, 3H), 1.76 (s, 3H), 1.71 (s, 3H); Mass: C21H21N5O [M+H] + Calculated: 360.2, Found: 360.2.

[0393] Example 147: Preparation of 3-(4-amino-6-isopropyl-7-methyl-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0394] The compound of Example 147 was prepared from the compound of Example 146 according to the hydrogenation method of Example 19 (8.5 mg, yield: 79%); 1HNMR (600 MHz, Methanol-d4) δ 8.57 (s, 1H), 8.21 (s, 1H), 7.08 (d, J = 8.3 Hz, 1H), 6.92 (d, J = 8.3 Hz, 1H), 3.36 - 3.32 (m, 1H), 2.60 (s, 3H), 1.75 (s, 3H), 1.70 (s, 3H), 1.40 (d, J = 6.9 Hz, 6H); Mass: C21H23N5O [M+H] + Calculated: 362.2, Found: 362.2.

[0395] Example 148: Preparation of 3-(4-amino-6-(2-hydroxypropan-2-yl)-7-methyl-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0396] The compound of Example 146 (1.0 eq), meso-tetraphenylporphyrin cobalt (1.0 eq), tetraethylammonium borohydride (10 eq) were added into a reaction bottle, then methanol and ethylene glycol dimethyl ether were added, oxygen was replaced for 3 times, and the reaction was stirred at 70 °C for 4 hours. TLC detection, after the reaction was completed, the pH was adjusted to 7 by adding aqueous sodium bicarbonate solution, EA extraction, the organic phase was washed with brine for 3 times, the organic phase was dried with anhydrous sodium sulfate, suction filtered, rotary evaporated, and separated and purified by reverse phase column chromatography (MeOH / H2O = 0.1-10) to obtain the compound of Example 148 (22 mg, yield: 22%); 1 H NMR (600 MHz, Methanol-d4) δ 8.84 (s, 1H), 8.42 (s, 1H), 7.10 (d, J = 8.3 Hz, 1H), 6.95 (d, J = 8.3 Hz, 1H), 2.86 (s, 3H), 1.78 (s, 6H), 1.75 (s, 3H), 1.69 (s, 3H); Mass: C21H23N5O2 [M+H] + Calculated: 378.2, Found: 378.2.

[0397] Example 149: Preparation of 3-(4-amino-6-(2-fluoropropan-2-yl)-7-methyl-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0398] The compound of Example 148 (1.0 eq) was dissolved in dichloromethane, and DAST (5 eq) was added dropwise under ice bath. The reaction was stirred for 5 hours under ice bath. TLC detection showed that the reaction was completed. After the reaction was completed, ammonium chloride solution was added dropwise for quenching, and EA extraction was performed. The organic phase was washed with brine for 3 times, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated and purified by reversed-phase column chromatography (MeOH / H2O = 0.1-10) to obtain the compound of Example 149 (5.3 mg, yield: 30%). 1 H NMR (600 MHz, Methanol-d4) δ 8.51 (d, J = 1.5 Hz, 1H), 8.13 (s, 1H), 7.01-6.97 (m, 1H), 6.83 (d, J = 8.3 Hz, 1H), 2.62 (s, 3H), 1.82 (s, 3H), 1.78 (s, 3H), 1.65 (s, 3H), 1.60 (s, 3H); Mass spectrum: C21H22N5OF [M+H] + Calculated: 380.2, Found: 380.2.

[0399] Example 150: Preparation of 2-(4-amino-9-(3-hydroxy-2,6-dimethylphenyl)-7-methyl-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-6-yl)-2-methylpropanenitrile

[0400] The compound of Example 148 (10 mg, 26.49 μmol) was dissolved in DCM (1 mL), and TMSCN (5.3 mg, 53 μmol, 6.63 μL) and InBr3 (939.30 μg, 2.65 μmol) in DCM (1 mL) were added under stirring at room temperature. The reaction was continued at room temperature for 1 hour. After the reaction solution was concentrated, the compound of Example 150 (0.6 mg, yield: 5% yield) was obtained by preparative high performance liquid chromatography, which was a white solid. 1 H NMR (600 MHz, Methanol-d4) δ 8.51 (d, J = 1.5 Hz, 1H), 8.13 (s, 1H), 7.01-6.97 (m, 1H), 6.83 (d, J = 8.3 Hz, 1H), 2.62 (s, 3H), 1.82 (s, 3H), 1.78 (s, 3H), 1.65 (s, 3H), 1.60 (s, 3H); Mass spectrum: C21H22N5OF [M+H] + Calculated: 387.2, Found: 387.2.

[0401] Example 151: Preparation of 3-(4-amino-6-(cyclopropylethynyl)-7-methyl-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0402] To a microwave reaction tube was added compound F-8 (20 mg, 1.0 eq), the corresponding terminal alkyne compound cyclopropylacetylene (1.5 eq), Pd(PPh3)4(0.1 eq), Cul (0.05 eq), triethylamine (8.0 eq), and anhydrous DMF (1 mL) as solvent, and the mixture was replaced with nitrogen and warmed to 120 °C for 1 h. After the reaction was completed, EA was added for extraction, and the organic phase was concentrated and purified by preparative high-performance liquid chromatography to obtain the compound of Example 151 (7.8 mg, yield: 27%). 1 H NMR (600 MHz, methanol-d4) δ 8.46 (s, 1H), 8.30 (s, 1H), 7.09 (d, J = 8.4 Hz, 1H), 6.94 (d, J = 8.3 Hz, 1H), 2.66 (s, 3H), 1.78 (s, 3H), 1.73 (s, 3H), 1.55 (m, 1H), 0.98-0.92 (m, 2H), 0.86-0.81 (m, 2H); Mass: C23H21N5O [M+H] + Calculated: 384.2, Found: 384.2.

[0403] Example 152: Preparation of 3-(4-amino-7-methyl-6-(pyrrolidin-1-yl)-9H-pyrrolo[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0404] Step 1: Synthesis of compound F-9

[0405] To a reaction bottle were added compound F-7 (1.0 eq), the corresponding amine (5 eq), tBuXPhos Pd G4 (20% eq), tBuXPhos (20% eq), and sodium tert-butoxide (3.0 eq), followed by anhydrous 1,4-dioxane. The mixture was replaced with nitrogen three times and stirred at 110 °C for 8 h. After the reaction was completed, the insoluble matter was filtered off, and the filtrate was extracted with water and EA. The organic phase was washed with brine three times, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was separated and purified by column chromatography (PE:EA, 0→100%) to obtain the product F-9.

[0406] Step 2: Synthesis of the compound of Example 152

[0407] Compound F-9 (1.0 eq) was dissolved in dichloromethane, and stirred at 0 °C, boron tribromide (5 eq) was added, and the reaction was allowed to proceed at room temperature for 3 h. The solvent was removed by concentration, and the pH was adjusted to 7 by adding aqueous sodium bicarbonate solution. The organic phase was extracted with ethyl acetate three times, combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by reverse phase column chromatography (MeOH / H2O = 0.1-10) to give the compound of Example 152 (7 mg, yield: 72%). 1 H NMR (600 MHz, methanol-d4) δ 8.68 (s, 1H), 8.43 (s, 1H), 7.10 (d, J = 8.3 Hz, 1H), 6.95 (d, J = 8.3 Hz, 1H), 3.57-3.51 (m, 4H), 2.67 (s, 3H), 2.22-2.14 (m, 4H), 1.75 (s, 3H), 1.69 (s, 3H); Mass: C22H24N6O [M+H] + Calculated: 389.2, Found: 389.2.

[0408] Examples 153-205:

[0409] The synthesis method of Reference Examples 143-152 was used to prepare the corresponding target compounds using compound F-8 and the starting materials listed in Table 9.

[0410] Table 9: Compounds of Examples 153-205

[0411] Example 206: Preparation of 4-amino-9-(3-hydroxy-2,6-dimethylphenyl)-6- (thiophen-2-yl)-9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidine-7-carbonitrile

[0412] Step 1: Synthesis of compound G-2

[0413] A reaction bottle was charged with G-1 (1.0 eq) and ammonium chloride (6.0 eq) and iron powder (3.0 eq) was added, followed by EtOH / H2O (5:1). The reaction mixture was heated to 80 °C and stirred for 1 h. The reaction was monitored by TLC. After the reaction was completed, H2O was added, followed by ethyl acetate. The mixture was filtered through celite, concentrated, and purified by column chromatography to give G-2 as a light yellow solid.

[0414] Step 2: Synthesis of compound G-3

[0415] In a reaction flask was added G-2 (1.0 eq) and concentrated sulfuric acid / water (1 / 5), ice water bath was lowered to 0 °C, then slowly added sodium nitrite (1.1 eq) and reacted for 1 h, stirred for 30 min, then warmed to 100 °C and stirred for 1 h, TLC monitoring, the reaction was completed, the reaction liquid was adjusted to PH to 6, EA extraction, organic layer was obtained, concentrated, column chromatography, light yellow solid G-3 was obtained.

[0416] Step 3: synthesis of compound G-4

[0417] In a reaction flask was added G-3 (1.0 eq) and silver carbonate (2.0 eq) in toluene as solvent, bromobenzyl (1.05 eq) was added dropwise in the suspension, stirred at room temperature for 3 h, TLC monitoring, the reaction was completed, diatomite was filtered, washed with ethyl acetate, concentrated, extracted with ethyl acetate and saturated sodium chloride solution, the organic layer was obtained, dried with anhydrous sodium sulfate, filtered, concentrated to obtain the target compound oil liquid G-4.

[0418] Step 4: synthesis of compound G-5

[0419] In a reaction flask was added G-4 (1.0 eq), 3-methoxy-2,6-dimethylaniline (1.05 eq), Pd2(dba)3 (0.05 eq), Xantphos (0.1 eq) and potassium tert-butoxide (2.0 eq), toluene as solvent, nitrogen replacement, warmed to 80 °C for 45 min, concentrated, dissolved in dichloromethane, silica gel was mixed, column chromatography to obtain the target compound oil liquid G-5.

[0420] Step 5: synthesis of compound G-6

[0421] In a reaction flask was added malononitrile (2.1 eq) and anhydrous ethylene glycol dimethyl ether as solvent, NaH (60% dispersion in oil) (3.0 eq) was added under ice water bath, stirred for 30 min, then G-5 (1.0 eq) and 1,1-bis(diphenylphosphino)ferrocene palladium dichloride (0.1 eq) were added, nitrogen replacement, warmed to 110 °C for 12 h, TLC monitoring, the reaction was completed, diatomite was filtered, washed with ethyl acetate, the filtrate was obtained, concentrated, column chromatography to obtain the target compound G-6.

[0422] Step 6: synthesis of compound G-7

[0423] In a reaction flask was added G-6 (1.0 eq) and tetrahydrofuran as solvent, NBS was added under ice water bath, warmed to room temperature, stirred for 3 h, TLC monitoring, the reaction was completed, diatomite was filtered, extracted with ethyl acetate, the organic layer was obtained, concentrated, column chromatography to obtain the target compound G-7.

[0424] Step 7: synthesis of compound G-8

[0425] In a reaction flask, G-7 (1.0 eq) was taken, then potassium ferrocyanide (1.0 eq) was added, tBuXPhos-Pd-G3 (0.1 eq) was added, 1,4-dioxane was used as a solvent, and a solution of potassium acetate (0.5 N) was added. The reaction was heated to 100 °C under N2protection for 2 h. After the reaction was completed, extraction was performed with EA to obtain an organic layer. Concentration, column chromatography, and concentration of the majority of methanol were performed to obtain the target compound G-8.

[0426] Step 8: Synthesis of compound G-9

[0427] In a reaction flask, G-8 (1.0 eq) was taken, and formamide was used as a reaction reagent and solvent. The reaction was heated to 205 °C for 40 min. After the reaction was completed, extraction was performed with dichloromethane to obtain an organic layer. Concentration, drying, and column chromatography were performed to obtain the target compound G-9.

[0428] Step 9: Synthesis of compound G-10

[0429] In a reaction flask, G-9 (1.0 eq) was taken, Pd / C (10% on Carbon) (0.15 eq) was added, and methanol was used as a solvent. The reaction was stirred at 60 °C under hydrogen protection for 4 h. After the reaction was completed, filtration was performed with diatomite, and washing was performed with methanol. Concentration of the filtrate was performed to obtain the target compound G-10.

[0430] Step 10: Synthesis of compound G-11

[0431] In a reaction flask, G-10 (1.0 eq) was taken, and THF was used for dissolution. N-phenyl bis(trifluoromethanesulfonimide) (1.0 eq) and triethylamine (2.0 eq) were added, and the reaction was stirred at room temperature for 1 h. After the reaction was completed, extraction was performed with dichloromethane to obtain an organic layer. Concentration, drying, and column chromatography were performed to obtain the target compound G-11.

[0432] Step 11: Synthesis of compound G-12

[0433] In a reaction flask, G-11 (1.0 eq) was taken, dichloromethane was used as a solvent, and boron tribromide (3.0 eq) was slowly added dropwise. The reaction was stirred at room temperature for 1 h. After the reaction was completed, methanol was added for quenching under an ice water bath. Concentration to remove excess solvent, reverse phase column chromatography, concentration of the majority of methanol, extraction with dichloromethane to obtain an organic layer, concentration, and vacuum drying were performed to obtain the target compound G-12.

[0434] Step 12: Synthesis of compound of Example 206

[0435] A microwave reaction tube was charged with G-12 (1.0 eq), the corresponding boronic acid or boronic ester compound 2-thiopheneboronic acid (1.5 eq), 1,1-bis(diphenylphosphino)ferrocene palladium dichloride (0.1 eq), potassium carbonate (2.0 eq), and dioxane and water (v:v = 2:1), replaced with N2, and heated to 110 °C for 1 h under N2protection. Upon completion, ethyl acetate and saturated sodium chloride solution were added to extract, and the organic layer was filtered, concentrated, and prepared to give the compound of Example 206 (9.2 mg, yield: 49.3%). 1 H NMR (400 MHz, Methanol-d4) δ 8.97 (s, 1H), 8.33 (s, 1H), 7.67 (d, J = 5.1 Hz, 1H), 7.61 (d, J = 3.6 Hz, 1H), 7.24 (dd, J = 5.1, 3.7 Hz, 1H), 7.12 (d, J = 8.3 Hz, 1H), 6.96 (d, J = 8.2 Hz, 1H), 1.80 (s, 3H), 1.74 (s, 3H); Mass spectrum: [M+H]+calculated for C22H17N6OS: 413.1, found: 413.2.

[0436] Examples 207-211:

[0437] The synthesis method of Reference Example 206 was used to prepare the corresponding target compound using compound G-12 and the starting materials listed in Table 10.

[0438] Table 10: Compounds of Examples 207-211

[0439] Example 212: Preparation of 3-(4-amino-7-(methyl-d3)-6-(thiophen-2-yl)-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0440] Step 1: Synthesis of compound H-1

[0441] A reaction bottle was charged with a solution of zinc chloride (0.5 mol / L in THF) (5.0 eq), replaced with N2, and then deuterated methyl magnesium bromide (1.0 mol / L in diethyl ether, 5.0 eq) was added. After stirring for 20 min, G-8 (1.0 eq) and Pd(PPh3)4(0.1 eq) were added, and the temperature was raised to 70 °C for stirring for 6 h. Upon completion, 1M HCl solution was added to quench, and EA was added to extract, and the organic layer was obtained. Column chromatography gave the product H-1.

[0442] Step 2: Synthesis of compound H-2

[0443] In a reaction flask, H-1 (1.0 eq) was taken and formamide was added as a reaction reagent and solvent, and the temperature was raised to 205 °C, and the reaction was carried out for 40 min, and the reaction was monitored by TLCMS, and after completion of the reaction, methylene chloride was added to extract the product, and the organic layer was concentrated, dried, and the crude product H-2 was obtained.

[0444] Step 3: Synthesis of compound H-3

[0445] In a reaction flask, H-2 (1.0 eq) and Pd / C (10% on Carbon, 0.15 eq) were taken, and methanol was added as a solvent, and hydrogen was passed, and the reaction was carried out for 4 h at 60 °C under hydrogen atmosphere, and after completion of the reaction, the reaction mixture was filtered through celite and washed with methanol, and the filtrate was concentrated to obtain the compound H-3.

[0446] Step 4: Synthesis of compound H-4

[0447] In a reaction flask, H-3 (1.0 eq) was taken, and THF was added to dissolve the compound, and N-phenyl bis(trifluoromethanesulfonimide) (1.0 eq) and triethylamine (2.0 eq) were added, and the reaction was carried out for 1 h at room temperature, and the reaction was monitored by TLCMS, and after completion of the reaction, methylene chloride was added to extract the product, and the organic layer was concentrated, dried, and column chromatography was carried out to obtain the compound H-4.

[0448] Step 5: Synthesis of compound H-5

[0449] In a reaction flask, H-4 (1.0 eq) was taken, and dichloromethane was added as a solvent, and boron tribromide (3.0 eq) was added dropwise slowly, and the reaction was carried out for 1 h at room temperature, and the reaction was monitored by TLC, and after completion of the reaction, methanol was added to quench the reaction in an ice water bath, and the excess solvent was removed by concentration, and the product was purified by reverse phase column chromatography, and the compound H-5 was obtained by concentrating the methanolic layer, extracting with dichloromethane, and drying under vacuum.

[0450] Step 6: Synthesis of compound of example 212

[0451] In a microwave reaction tube, H-5 (1.0 eq), the corresponding boronic acid or boronic ester compound 2-thiopheneboronic acid (1.5 eq), 1,1-bis(diphenylphosphino)ferrocene palladium dichloride (0.1 eq), potassium carbonate (2.0 eq), and dioxane and water (v:v = 2:1) were taken, and N2was passed, and the reaction was carried out for 1 h at 110 °C under N2, and after completion of the reaction, ethyl acetate and saturated sodium chloride solution were added to extract the product, and the organic layer was filtered, concentrated, and purified by preparative HPLC to obtain the compound of example 212 (3.3 mg, yield: 67%); 1H NMR (400 MHz, Methanol-d4) δ 8.88 (s, 1H), 8.45 (s, 1H), 7.57 (dd, J = 5.2, 1.2 Hz, 1H), 7.26 (dd, J = 3.5, 1.2 Hz, 1H), 7.19 (dd, J = 5.1, 3.6 Hz, 1H), 7.12 (d, J = 8.3 Hz, 1H), 6.96 (d, J = 8.3 Hz, 1H), 1.80 (s, 3H), 1.73 (s, 3H); Mass: C22H17D3N5OS [M+H] + Calculated: 405.2, Found: 405.2.

[0452] Examples 213-215:

[0453] The synthesis method of Reference Example 212 was used to prepare the corresponding target compound using compound H-5 and the starting materials listed in Table 11.

[0454] Table 11: Compounds of Examples 213-215

[0455] Example 216: Preparation of 3-(4-amino-6-(thiophen-2-yl)-7-(trifluoromethyl)- 9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0456] Step 1: Synthesis of compound I-1

[0457] Into a reaction bottle was added G-6 (1.0 eq), tetrahydrofuran was added as solvent, NIS (1.1 eq) was added under ice water bath, the temperature was increased to room temperature, the reaction was stirred for 3 h, TLC monitoring, the reaction was completed, diatomite was filtered, ethyl acetate was extracted, the organic layer was concentrated, column chromatography was carried out, and the target compound I-1 was obtained.

[0458] Step 2: Synthesis of compound I-2

[0459] Into a reaction bottle was added I-1 (1.0 eq) and trifluoromethyl (1,10- phenanthroline) copper (I) (2.0 eq), anhydrous DMF was used as solvent, the temperature was increased to 70°C, the reaction was stirred for 4 h, the reaction was completed and filtered, EA was extracted, the organic layer was concentrated, and column chromatography was carried out to obtain the product I-2.

[0460] Step 3: Synthesis of compound I-3

[0461] In a reaction flask, I-2 (1.0 eq) was taken as a reaction reagent and solvent, and warmed to 205 °C for 40 minutes. The reaction was monitored by TLCMS. After the reaction was completed, dichloromethane was added for extraction. The organic layer was concentrated, dried, and the target compound I-3 was obtained.

[0462] Step 4: Synthesis of compound I-4

[0463] In a reaction flask, I-3 (1.0 eq) was taken, Pd / C (10% on Carbon, 0.15 eq) was added, and methanol was added as a solvent. Hydrogen was replaced, and stirring was performed under hydrogen protection at 60 °C for 4 hours. After the reaction was completed, diatomite was filtered, washed with methanol, and the filtrate was concentrated to obtain the target compound I-4.

[0464] Step 5: Synthesis of compound I-5

[0465] In a reaction flask, I-4 (1.0 eq) was taken, dissolved in THF, and N-phenyl bis(trifluoromethylsulfone) imine (1.0 eq) and triethylamine (2.0 eq) were added. Stirring was performed at room temperature for 1 h, and the reaction was monitored by TLCMS. After the reaction was completed, dichloromethane was added for extraction. The organic layer was concentrated, dried, and column chromatography was performed to obtain the target compound I-5.

[0466] Step 6: Synthesis of compound I-6

[0467] In a reaction flask, I-5 (1.0 eq) was taken, dichloromethane was added as a solvent, and boron tribromide (3.0 eq) was slowly added dropwise. Stirring was performed at room temperature for 1 h, and the reaction was monitored by TLC. After the reaction was completed, methanol was added for quenching under an ice water bath. The excess solvent was removed by concentration, and reverse phase column chromatography was performed. Most of the methanol was concentrated, dichloromethane was added for extraction, and the organic layer was concentrated and dried under vacuum to obtain the target compound I-6.

[0468] Step 7: Synthesis of compound of Example 216

[0469] In a microwave reaction tube, I-6 (1.0 eq), the corresponding boronic acid or boronic ester compound 2-thiopheneboronic acid (1.5 eq), 1,1-bis(diphenylphosphino)ferrocene palladium dichloride (0.1 eq), potassium carbonate (2.0 eq), and dioxane and water (v:v = 2:1) were added. N2 was replaced, and the temperature was increased to 110 °C for 1 h under microwave protection. After the reaction was completed, ethyl acetate and saturated sodium chloride solution were added for extraction. The organic layer was filtered, concentrated, and HPLC preparation was performed to obtain the compound of Example 214 (10.8 mg, yield: 45%); 1H NMR (600 MHz, Methanol-d4) δ 8.86 (s, 1H), 8.32 (s, 1H), 7.67 (s, 1H), 7.56 (d, J = 8.0 Hz, 2H), 7.21 - 7.10 (m, 2H), 6.95 (d, J = 8.1 Hz, 1H), 1.77 (s, 3H), 1.73 (s, 3H); Mass: [M+H]+calcd for C22H17F3N5OS: 456.1, found: 456.1.

[0470] Examples 217-221:

[0471] The synthesis method of Reference Example 216 was used to prepare the corresponding target compound using compound I-6 and the starting materials listed in Table 12.

[0472] Table 12: Compounds of Examples 217-221

[0473] Example 222: Preparation of 3-(4-amino-7-methyl-6-(1H-pyrrol-1-yl)-9H-pyrrolo[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0474] Step 1: Synthesis of compound J-1

[0475] Compound F-5 (1.5 g, 2.86 mmol) was dissolved in DMF (22.5 mL), potassium carbonate (2.59 g, 18.7 mmol) was added, and bromopropene (0.87 g, 7.15 mmol) was added. The reaction was reacted at 80 degrees for 2 hours, potassium carbonate (1.19 g, 8.65 mmol) was added, bromopropene (0.44 g, 2.88 mmol) was added, and the reaction was reacted at 80 degrees for 2 hours. After the reaction was completed, it was diluted with ethyl acetate, washed with saturated brine three times, dried with anhydrous ammonium sulfate, rotary evaporated, and separated and purified by column chromatography (PE / EA = 11:1) to obtain 0.9 g of product J-1 with a yield of 52%.

[0476] Step 2: Synthesis of compound J-2

[0477] Compound 2A (200 mg, 0.33 mmol) was dissolved in DCM (3 mL), Grubbs second-generation catalyst (14.0 mg, 0.017 mmol) was added, and the reaction was reacted at room temperature for 2 hours. After the reaction was completed, it was rotary evaporated, and separated and purified by column chromatography (PE / EA = 10:1) to obtain 0.14 g of product J-2 with a yield of 74%.

[0478] Step 3: Synthesis of compound J-3

[0479] Compound J-2 (0.14 g, 0.24 mmol) was dissolved in formamide (2 mL), the reaction was carried out in the heating module 205 degree for 50 minutes, after the reaction was completed, diluted with ethyl acetate, washed with saturated brine three times, dried with anhydrous ammonium sulfate, rotary evaporation, column chromatography (PE / EA = 1:1) to separate and purify 17 mg of product J-3, the yield was 18%.

[0480] Step 4: Synthesis of the compound of Example 222

[0481] Compound J-3 (17 mg, 0.043 mmol) was dissolved in DCM (0.2 mL), cooled to 0 degrees, and BBr3 (1 M, 0.43 mL) was added dropwise. The reaction temperature was raised to 50 degrees, and the reaction was carried out at this temperature for 30 minutes. After the reaction was completed, it was cooled to room temperature, and the reaction solution was slowly added to saturated sodium bicarbonate. An equal volume of DCM was added to the saturated sodium bicarbonate, filtered, and the filter cake was washed with water three times. The solid was collected, the filtrate was separated and dried, rotary evaporation, and the filter cake was washed with DCM (1 mL) for 2 hours, filtered, and the filter cake was washed with DCM three times. Rotary evaporation under reduced pressure gave the compound of Example 220 (3 mg, yield: 16%); 1 H NMR (400 MHz, CD3OD) δ 8.53 (s, 1H), 8.15 (s, 1H), 7.02-7.00 (m, 1H), 6.85-6.82 (m, 3H), 6.23 (s, 2H), 2.27 (s, 3H), 1.69 (s, 3H), 1.63 (s, 3H); Mass spectrum: C25H27N4S [M+H] + Calculated: 385.2, Found: 385.2.

[0482] Example 223: Preparation of 3-(4-amino-6-(diallylamino)-7-methyl-9H-pyrrolo[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0483] The target molecule was prepared according to the synthesis method of Reference Example 220 (6 mg, yield: 38%). 1 H NMR (400 MHz, CD3OD) δ 8.53 (s, 1H), 8.15 (s, 1H), 7.02-7.00 (m, 1H), 6.85-6.82 (m, 3H), 6.23 (s, 2H), 2.27 (s, 3H), 1.69 (s, 3H), 1.63 (s, 3H); Mass spectrum: C25H27N4S [M+H] +Calculated: 415.2, Found: 415.2.

[0484] Example 224: Preparation of 1-(4-amino-9-(3-hydroxy-2,6-dimethylphenyl)-7- methyl-9H-pyrrolo[3',2':4,5]pyrrolo[2,3-d]pyrimidin-6-yl)pyrrolidin-2-one

[0485] Step 1: Synthesis of compound J-4

[0486] Compound F-5 (770 mg, 1.48 mmol, 1.0 eq.), chlorobutyryl chloride (0.17 mL, 1.48 mmol, 1.0 eq.), potassium carbonate (613 mg, 4.44 mmol, 3.0 eq.), dichloromethane (1.48 mL, 1M) were added into a reaction bottle, stirred at room temperature; after 2 hours of reaction, the reaction was monitored by spotting plate, diluted with water, extracted with EA twice, dried over anhydrous sodium sulfate, and concentrated by rotary evaporation; purified by silica gel column chromatography to obtain compound J-4 (500 mg, yield 59%); MS [M+1]=626.

[0487] Step 2: Synthesis of compound J-5

[0488] Compound J-4 (500 mg, 0.8 mmol, 1.0 eq.), potassium tert-butoxide (136 mg, 1.2 mmol, 2.0 eq.), acetonitrile (8 mL, 0.1M) were added into a reaction bottle; stirred at room temperature until the reaction was monitored by spotting plate, diluted with water, extracted with EA twice, dried over anhydrous sodium sulfate, and concentrated by rotary evaporation; purified by column chromatography to obtain compound J-5 (250 mg, yield 53%); MS [M+1]=590.

[0489] Synthesis of the compound of Example 224

[0490] Step 3-4 was prepared according to the method for synthesizing the compound of Example 220 (2 mg, yield: 12%); 1 H NMR (400 MHz, Methanol-d4) δ 8.62 (s, 1H), 8.25 (s, 1H), 7.09 (d, J = 8.3 Hz, 1H), 6.93 (d, J = 8.3 Hz, 1H), 3.94 (t, J = 7.0 Hz, 2H), 2.65 (t, J = 8.1 Hz, 2H), 2.47 (s, 3H), 2.35 (t, J = 7.5 Hz, 2H), 1.76 (s, 3H), 1.70 (s, 3H); Mass spectrum: C22H22N6O2, [M+H] + Calculated: 403.2, Found: 403.2.

[0491] Example 225: Preparation of 3-(4-amino-7-(difluoromethyl)-6-(thiophen-2-yl)- 9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0492] Step 1: Synthesis of compound K-1

[0493] Into a reaction vial, was placed G-11 (90 mg, 182.8 μmol), 2-thiopheneboronic acid (35.1 mg, 274.2 μmol), 1,1-bis(diphenylphosphino)ferrocene palladium dichloride (13.4 mg, 18.3 μmol), potassium carbonate (75.8 mg, 548.3 μmol), H2O (1 mL) and 1,4-dioxane (2 mL), the vial was replaced with N2, and the reaction was stirred at 110 °C for 40 min. The reaction was completed, extracted with ethyl acetate, the organic layer was concentrated, and purified by column chromatography to give 40 mg of intermediate compound K-1 in 51% yield.

[0494] Step 2: Synthesis of compound K-2

[0495] Into a reaction vial, was placed K-1 (40 mg, 93.8 μmol) and anhydrous tetrahydrofuran (2 mL) to dissolve, then DIBAL-H (1.0 M in heptane, 468.9 μmol) was added, and the reaction was stirred at room temperature for 3 h. The reaction was completed, quenched with 0.5 mL of ammonium chloride solution, and then the pH was adjusted to basic with sodium bicarbonate solution. The reaction was extracted with ethyl acetate, the organic layer was concentrated, and 24 mg of intermediate product K-2 was prepared in 24% yield.

[0496] Step 3: Synthesis of compound K-3

[0497] Into a reaction vial, was placed K-2 (10 mg, 23.3 μmol) and dichloromethane (2 mL) to dissolve, then DAST (18.8 mg, 116.4 μmol) was added, and the reaction was stirred at room temperature for 3 h. The reaction was completed, quenched with sodium bicarbonate solution, extracted with dichloromethane, and the aqueous phase was extracted with ethyl acetate again. The organic layers were combined, concentrated, and 2 mg of product K-3 was prepared in 24% yield.

[0498] Step 4: Synthesis of Example 225 compound

[0499] A reaction vial was charged with K-3 (2 mg, 4.4 pmol), dissolved in dichloromethane (2 mL), and boron tribromide (13.3 pmol) was added dropwise. The reaction was stirred at room temperature for 2 h. Upon completion, the reaction was quenched with methanol in an ice water bath, followed by the addition of saturated sodium bicarbonate solution. The aqueous phase was extracted with dichloromethane, and the organic layers were combined and concentrated. The residue was purified by preparative HPLC to give the compound of Example 225 (0.7 mg, 35% yield); 1 H NMR (400 MHz, Methanol-d4) δ 8.84 (s, 1H), 8.30 (s, 1H), 7.60 - 7.63 (m, 1H), 7.24 - 7.26 (m, 1H), 7.18 - 7.21 (m, 1H), 7.11 (d, J = 8.3 Hz, 1H), 6.95 (d, J = 8.2 Hz, 1H), 6.62 - 6.89 (m, 1H), 1.80 (s, 3H), 1.74 (s, 3H); Mass spectrum: C22H18F2N5OS [M+H] + Calculated: 438.1, Found: 438.2.

[0500] Example 226: Preparation of 3-(4-amino-7-(difluoromethyl)-6-(pyrrolidin-1-yl)- 9H-pyrrolo[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0501] Step 1: Synthesis of compound K-4

[0502] A reaction vial was charged with G-2 (3.8 g, 15.1 mmol), 1,4-diiodobutane (9.4 g, 30.2 mmol), and EtOH / H2O (20 ml, 1 / 1), and the reaction was stirred at 100 °C for 48 h. Upon completion, the reaction was quenched with saturated sodium chloride solution, and the organic layer was extracted with ethyl acetate. The residue was purified by column chromatography to give 1.2 g of compound K-4 in 26% yield.

[0503] Step 2: Synthesis of compound K-5

[0504] A microwave reaction tube was charged with K-4 (1 g, 3.27 mmol), 2,6-dimethyl-3- methoxyaniline (494.1 mg, 3.27 mmol), Pd2(dba)3 (149.6 mg, 163.40 pmol), Xantphos (189.1 mg, 326.8 pmol), sodium tert-butoxide (628.1 mg, 6.54 mmol), and toluene (15 mL) as solvent. The reaction was stirred at 110 °C for 1 h. Upon completion, the reaction was filtered through celite, and the filtrate was concentrated and purified by column chromatography to give 1.0 g of the target compound K-5 in 81% yield.

[0505] Step 3: Synthesis of compound K-6

[0506] Into a microwave vial was added malononitrile (368.7 mg, 5.6 mmol), ethylene glycol dimethyl ether (15 mL) as solvent, sodium hydride (60% dispersion in oil, 191.3 mg, 8.0 mmol) was added and stirred for 5 min, K-5 (1.0 g, 2.66 mmol) was added, 1,1-bis(diphenylphosphino)ferrocene palladium dichloride (291.6 mg, 398.6 μmol) was added, N2 was replaced, and the temperature was increased to 110 °C for 1.5 h under microwave. TLC was used to monitor the reaction. After the reaction was completed, the reaction mixture was filtered through celite, and the filtrate was concentrated and purified by column chromatography to give 500 mg of the target compound K-6 with a yield of 52%.

[0507] Step 4: Synthesis of compound K-7

[0508] Into a reaction vial was added K-6 (500 mg, 1.4 mmol), N,N-dimethylformamide (15 mL) as solvent, and the temperature was cooled to 0 °C. NBS (270.8 mg, 1.5 mmol) was added in three portions with an interval of 10 min. The temperature was slowly increased to room temperature and stirred overnight. After the reaction was completed, saturated sodium thiosulfate solution was added, and ethyl acetate was used to extract the organic layer. The organic layer was concentrated and purified by column chromatography to give 320 mg of the target compound K-7 with a yield of 52%.

[0509] Step 5: Synthesis of compound K-8

[0510] Into a reaction vial was added K-7 (120 mg, 272.5 μmol), potassium ferrocyanide (100.4 mg, 272.5 μmol), tBuXPhos G3 Pd (21.6 mg, 27.3 μmol), potassium acetate (53.5 mg, 545.0 μmol), 1,4-dioxane (5 mL), and H2O (1 mL) as solvent. N2 was replaced, and the temperature was increased to 100 °C for 2 h under N2 protection. After the reaction was completed, ethyl acetate was used to extract the organic layer. 100 mg of the crude product K-8 was obtained with a yield of 85%.

[0511] Step 6: Synthesis of compound K-9

[0512] Into a reaction vial was added K-8 (90 mg, 232.9 μmol), formamide (5 mL), and the temperature was increased to 190 °C for 40 min. After the reaction was completed, saturated sodium chloride solution was added, and ethyl acetate was used to extract the organic layer. The organic layer was concentrated and purified by column chromatography to give 70 mg of the product K-9 with a yield of 72%.

[0513] Step 7: Synthesis of compound K-10

[0514] A reaction vial was charged with K-9 (50 mg, 120.9 μmol), anhydrous tetrahydrofuran (3 mL) was added to dissolve, diisopropylaluminum hydride (1.0 M, 605 μL) was added under ice-salt bath, the reaction was stirred at room temperature for 3 h, the reaction was completed, ammonium chloride solution (0.5 mL) was added to quench, sodium bicarbonate solution was added to adjust to basic, ethyl acetate was added to extract, the organic layer was obtained, concentrated, and 15 mg of product K-10 was prepared with a yield of 30%.

[0515] Step 8: Synthesis of compound K-11

[0516] A reaction vial was charged with K-10 (15 mg, 36.02 μmol), dichloromethane was added to dissolve, DAST (11.61 mg, 72.03 μmol) was added under 0 °C, the reaction was stirred for 3 h, the reaction was monitored, the reaction was completed, ammonium chloride solution was added to quench, dichloromethane was added to extract, the organic layer was obtained, and 8 mg of product K-11 was prepared with a yield of 50%.

[0517] Step 9: Synthesis of compound of Example 226

[0518] A reaction vial was charged with K-11 (8 mg, 18.6 μmol), dichloromethane (2 mL) was added to dissolve, boron tribromide (54.7 μmol) was added dropwise, the reaction was stirred at room temperature for 2 h, the reaction was monitored, the reaction was completed, methanol was added dropwise under ice-water bath to quench, sodium bicarbonate solution was added to extract, the aqueous phase was extracted with ethyl acetate again, the organic layers were combined, concentrated, and HPLC preparation and purification yielded compound of Example 226 (5.0 mg, yield: 61%); 1 H NMR (400 MHz, Methanol-d4) δ 8.68 (s, 1H), 8.47 (s, 1H), 7.23-6.93 (m, 3H), 3.41-3.34 (m, 4H), 2.12-2.03 (m, 4H), 1.77 (s, 3H), 1.71 (s, 3H); Mass spectrum: C22H23F2N6O [M+H] + Calculated: 425.2, Found: 425.2.

[0519] Example 227: Preparation of 4-amino-9-(3-hydroxy-2,6-dimethylphenyl)-6- (pyrrolidin-1-yl)-9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidine-7-carbonitrile

[0520] The compound of Example 227 was prepared according to the synthetic method of Reference Example 226 (5.0 mg, yield: 26%); 1H NMR (400 MHz, Methanol-d4) δ 8.44 (s, 1H), 8.25 (s, 1H), 7.10 (d, J = 8.3 Hz, 1H), 6.94 (d, J = 8.3 Hz, 1H), 3.65 - 3.74 (m, 4H), 2.15 - 2.06 (m, 4H), 1.79 (s, 3H), 1.72 (s, 3H); Mass: C22H22N7O [M+H] + Calculated: 400.2, Found: 400.2.

[0521] Example 228: Preparation of 3-(4-amino-7-(methyl-d3)-6-(pyrrolidin-1-yl)-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0522] Step 1: Synthesis of compound K-12

[0523] Deuterated methyl magnesium bromide (1.0 mol / L in ethyl ether, 135.6 μmol) and dioxane (2 mL) were added into a reaction flask at room temperature, then ZnCl2(0.5 mol / L in tetrahydrofuran, 678.19 μmol) was added, N2protected, stirred at room temperature for 20 min, a solution of K-7 (60 mg, 135.6 μmol) and Pd(PPh3)4(15.7 mg, 13.56 μmol) in dioxane (2 mL) was added into the system, N2protected, stirred at 150 °C overnight, TLC monitored the reaction, the reaction was completed, cooled to room temperature, 1M HCl was added for quenching, extracted with ethyl acetate and saturated brine, concentrated, purified by column chromatography (PE:DCM = 1:4) to give the product K-12 (40 mg, 77.9%).

[0524] Step 2: Synthesis of compound K-13

[0525] K-12 (40 mg, 105.6 μmol) was dissolved in formamide (2 mL), stirred at 200 °C for 1 hr, TLC monitored, the reaction was completed, cooled to room temperature, extracted with water and EA, washed with saturated brine, the organic phase was dried over anhydrous sodium sulfate, concentrated to give the crude product K-13 (20 mg, 46.7%).

[0526] Step 3: Synthesis of Example 228 compound

[0527] K-13 (20 mg, 49.3 μmo) was dissolved in DCM (2 mL), BBr3 (295.9 μmol) was added at 0 °C, the reaction was continued at 40 °C overnight, TLC was used to monitor the reaction, after the reaction was completed, methanol was added to quench the reaction, concentrated, and the compound of Example 228 (5.5 mg, yield: 28.2%) was obtained by acid HPLC preparation; 1 H NMR (400 MHz, DMSO-d6) δ 9.55 (s, 1H), 8.51 (s, 1H), 8.48 (s, 2H), 8.41 (s, 1H), 7.06 (d, J = 8.3 Hz, 1H), 6.94 (d, J = 8.2 Hz, 1H), 3.31 - 3.25 (m, 4H), 2.03 - 1.93 (m, 4H), 1.67 (s, 3H), 1.58 (s, 3H); Mass: C22H21D3N6O [M+H] + Calculated: 392.2, Found: 392.2.

[0528] Example 229: Preparation of 3-(4-amino-7-methyl-6-(1-(trifluoromethyl)cyclopropyl)- 9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0529] Step 1: Synthesis of compound L-1

[0530] Compound F-7 (300 mg, 0.73 mmol, 1.0 eq), isopropenylboronic acid pinacol ester ((200 mg, 1.8 mmol, 1.5 eq), Pd(dppf)Cl2(106 mg, 0.14 mmol, 20% eq), and potassium carbonate (300 mg, 2.18 mmol, 3.0 eq) were added into a reaction flask, then 1,4-dioxane (3 ml) and water (0.5 ml) were added, and the reaction was stirred at 120 °C for 2 hours under nitrogen. After the reaction was completed, the insoluble matter was filtered off, water and EA were added for extraction, the organic phase was washed with brine 3 times, the organic phase was dried over anhydrous sodium sulfate, and then filtered, concentrated, and separated and purified by silica gel column chromatography (PE / EA = 5 / 5) to obtain compound L-1 (200 mg, 64.3%).

[0531] Step 2: Synthesis of compound L-2

[0532] Compound L-1 (200 mg, 0.46 mmol, 1 eq) was dissolved in 1,4-dioxane, potassium hydroxide aqueous solution (2 ml, 35.6 mmol) was added, N-methyl-N-nitroso-p-toluenesulfonamide was added at room temperature, and the reaction was carried out at 65 °C for several hours. TLC detection was performed, and after the reaction was completed, TFA was added for quenching, sodium bicarbonate aqueous solution was added to adjust the pH to 7, most of the 1,4-dioxane was removed by concentration, extraction was performed with ethyl acetate three times, the organic phase was combined, dried with anhydrous sodium sulfate, filtered, and concentrated. Purification was performed by silica gel column chromatography (PE / EA = 2 / 8) to obtain the product L-2 (100 mg, 45.5%).

[0533] Step 3: Synthesis of compound L-3

[0534] Compound L-2 (100 mg, 0.21 mmol, 1 eq) and 1,2-dichlorobenzene were added to a microwave tube, and microwave heating was performed at 230 °C for 3 hours. TLC detection was performed, and after the reaction was completed, direct purification was performed by silica gel column chromatography (PE / EA = 5 / 5) to obtain the product L-3 (45 mg, 47.8%).

[0535] Step 4: Synthesis of the compound of Example 229

[0536] Compound L-3 (40 mg, 0.09 mmol, 1 eq) was dissolved in dichloromethane, and boron tribromide (227.0 mg, 0.90 mmol, 10 eq) was added at room temperature, and the reaction was carried out for 3 hours. TLC detection was performed, and after the reaction was completed, the solvent was removed by concentration, sodium bicarbonate aqueous solution was added to adjust the pH to 7, extraction was performed with ethyl acetate three times, the organic phase was combined, dried with anhydrous sodium sulfate, filtered, concentrated, and purified by reverse phase column chromatography (MeOH / H2O = 0.1-10) to obtain the product, the compound of Example 229 (14.0 mg, 68.8%). 1 H NMR (600 MHz, methanol-d4) δ 8.71 (s, 1H), 8.13 (s, 1H), 6.98 (d, J = 8.3 Hz, 1H), 6.82 (d, J = 8.3 Hz, 1H), 2.58 (s, 3H), 1.65 (s, 3H), 1.60 (s, 3H), 1.50 (m, 2H), 1.18 (m, 2H); Mass spectrum: C22H20N5OF3 [M+H] + Calculated: 428.2, Found: 428.2.

[0537] Example 230: Preparation of 3-(4-amino-7-cyclopropyl-6-(thiophen-2-yl)-9H-pyrrolo[2',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0538] Step 1: Synthesis of compound M-1

[0539] Under N2 atmosphere, G-7 (600 mg, 1.26 mmol), Pd(dppf)Cl2 (91.97 mg, 125.69 μmol), K2CO3 (521.2 mg, 3.77 mmol), and cyclopropylboronic acid (162 mg, 1.89 mmol) were added to a reaction flask. Dioxane (5 mL) and H2O (1 mL) were added, and the atmosphere was replaced with N2. The reaction was heated to 100°C for 1 hour, cooled to room temperature, and monitored. After completion of the reaction, the product was concentrated and purified by column chromatography (MeOH / DCM = 1 / 10) to afford product M-1 (520 mg, 94.3%).

[0540] Step 2-Step 6: Prepare Example 230 (6.5 mg, yield: 35.7%) by referring to the synthesis method of Example 206; 1 HNMR (400MHz, DMSO-d6) δ9.44(s,1H),8.79(s,1H),8.21(s,1H),7.69(dd,J=5.1,1.2Hz,1H),7.47(s,2H),7.32(dd,J=3.5,1.2Hz,1H),7.22(dd,J=5 .2,3.5Hz,1H),7.04(d,J=8.3Hz,1H),6.90(d,J=8.2Hz,1H),2.34–2.26(m ,1H),1.68(s,3H),1.59(s,3H),0.88–0.76(m,4H); Mass spectrum: C24H21N5OS; [M+H] + Calculated value: 428.2, measured value: 428.2.

[0541] Examples 231-240:

[0542] Refer to the synthetic method of Example 230 and use the starting materials listed in Table 13 to prepare the corresponding target compounds.

[0543] Table 13: Compounds of Examples 231-240

[0544] Example 241: Preparation of 3-(4-amino-8-methyl-7-(prop-1-en-2-yl)pyrrolo[1,5-a:3,2-d']dipyrimidin-10-yl)-2,4-dimethylphenol

[0545] Step 1: Synthesis of Compound N-1

[0546] Compound 5-bromo-4-methylpyrimidin-2-amine (3.00 g, 15.9 mmol) and allylboronic acid pinacol ester (3.21 g, 19.1 mmol) were dissolved in dioxane (30 mL) and water (3 mL), potassium carbonate (6.62 g, 47.9 mmol) and Pd(dppf)Cl2(1.17 g, 1.59 mmol) were added. The reaction was carried out at 100 degrees under N2protection for 15 hours, after the reaction was completed, concentrated, separated and purified by column chromatography (PE / EA = 3:1) to obtain 2.00 g of product N-1, with a yield of 84%.

[0547] Step 2: Synthesis of compound N-2

[0548] Compound N-1 (2.00 g, 13.4 mmol) was dissolved in acetonitrile (20 mL), copper chloride (2.34 g, 17.4 mmol) was added, and tert-butyl nitrite (2.07 g, 20.1 mmol) was added. The reaction was carried out at 75 degrees under N2protection overnight, after the reaction was completed, concentrated, separated and purified by column chromatography (PE / EA = 10:1) to obtain 0.42 g of product N-2, with a yield of 19%.

[0549] Step 3: Synthesis of compound N-3

[0550] Compound N-2 (0.42 g, 2.49 mmol) and 2,6-dimethyl-3-methoxyphenylacetonitrile (reference patent US20230122909 preparation method, 0.52 g, 2.99 mmol) were dissolved in tetrahydrofuran (5 mL), KHMDS (1M, 3.0 mL) was added dropwise, the reaction was carried out for 120 minutes, after the reaction was completed, the reaction solution was slowly added to 2M HCl, extracted with ethyl acetate, washed with saturated brine, and the crude product was separated and purified by column chromatography (PE / EA = 8:1) to obtain 0.38 g of product N-3, with a yield of 49%.

[0551] Step 4: Synthesis of compound N-4

[0552] Compound N-3 (0.38 g, 1.23 mmol) was dissolved in DMSO (5 mL), potassium carbonate (1.10 g, 7.98 mmol) was added, and bromoacetonitrile (0.37 g, 3.07 mmol) was added. The reaction was carried out at room temperature for 15 hours, after the reaction was completed, quenched with water, extracted with ethyl acetate, washed with saturated brine, and the crude product was separated and purified by column chromatography (PE / EA = 7:1) to obtain 0.25 g of product N-4, with a yield of 59%.

[0553] Step 5: Synthesis of compound N-5

[0554] Compound N-4 (0.25 g, 0.72 mmol) was dissolved in formamide (3 mL) and reacted for 50 min at 205 degree in a heating module. After the reaction was completed, it was diluted with ethyl acetate, washed with saturated brine three times, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated and purified by column chromatography (PE / EA = 1 : 1) to obtain 150 mg of product N-5 at a yield of 56%.

[0555] Step 6: Preparation of the compound of Example 241

[0556] Compound N-5 (150 mg, 0.40 mmol) was dissolved in DCM (2 mL), cooled to 0 degree, and BBr3 (1 M, 2.0 mL) was added dropwise. The reaction was reacted at room temperature for 2 hours. After the reaction was completed, the reaction solution was slowly added to a saturated sodium bicarbonate solution, and an equal volume of DCM was added. The mixture was filtered, the filter cake was washed with water three times, and the solid was collected. The filtrate was separated, dried, and concentrated under reduced pressure. The residue was slurried with DCM (1 mL) for 2 hours, filtered, and the filter cake was washed with DCM three times. The filtrate was concentrated under reduced pressure to obtain 90 mg of the compound of Example 241 at a yield of 63%. 1 H NMR (400 MHz, DMSO-d6) δ 8.99 (s, 1H), 8.24 (s, 1H), 6.97 (d, J = 8.2 Hz, 1H), 6.78 (d, J = 8.2 Hz, 1H), 5.44 (s, 1H), 5.18 (s, 1H), 2.54 (s, 3H), 2.18 (s, 3H), 1.71 - 1.99 (m, 6H); Mass: C21H22N5O [M+H] + Calculated: 360.2, Found: 360.2.

[0557] Example 242: Preparation of 3-(4-amino-7-isopropyl-8-methylpyrrolo[l,5-a:3,2-d']dipyrimidin-10-yl)-2,4-dimethylphenol

[0558] The compound of Example 241 (60 mg, 0.17 mmol) was dissolved in methanol (2 mL), 10% wet Pd / C (6 mg) was added, and the mixture was reacted under hydrogen gas at room temperature for 15 hours. After the reaction was completed, the mixture was filtered, concentrated, and separated and purified by TLC plate (DCM / MeOH = 10: 1) to obtain 20 mg of the compound of Example 242 at a yield of 33%. 1H NMR (400 MHz, DMSO-d6) δ 8.83 (s, 1H), 8.22 (s, 1H), 6.98 (d, J = 8.0 Hz, 1H), 6.76 (d, J = 8.0 Hz, 1H), 3.23-3.27 (m, 1H), 2.60 (s, 3H), 1.89 (s, 3H), 1.86 (s, 3H), 1.44 (d, J = 8.0 Hz, 6H); Mass: C21H24N5O [M+H] + Calculated: 362.2, Found: 362.2.

[0559] Example 243: Preparation of 3-(4-amino-8-methyl-7-(pyrrolidin-1-yl)pyrrolo[1,5- a:3,2-d']dipyrimidin-10-yl)-2,4-dimethylphenol

[0560] Step 1: Synthesis of compound N-6

[0561] Compound 2-chloro-4-methyl-5-aminopyrimidine (1.0 g, 6.96 mmol) and compound 1,4-diiodobutane (3.24 g, 10.44 mmol) were dissolved in DMF (10 mL), potassium carbonate (6.24 g, 45.24 mmol) was added. The reaction was reacted at 80 degrees under N2environment for 15 hours, after the reaction was completed, extracted with ethyl acetate, washed with saturated brine, the crude product was separated and purified by column chromatography (PE / EA = 9:1) to obtain 310 mg of product N-6, the yield was 23%.

[0562] Step 2-5: Refer to the synthesis method of Example 241 Step 3-6 to prepare the compound of Example 243

[0563] 1 H NMR (400 MHz, DMSO-d6) δ 8.56 (s, 1H), 8.18 (s, 1H), 6.96 (d, J = 8.0 Hz, 1H), 6.75 (d, J = 8.0 Hz, 1H), 3.23-3.25 (m, 4H), 2.58 (s, 3H), 2.03-2.05 (m, 4H), 1.89 (s, 3H), 1.86 (s, 3H); Mass: C22H25N6O [M+H] + Calculated: 389.2, Found: 389.2.

[0564] Examples 244-245:

[0565] Refer to the synthesis method of Example 241 to prepare, using the starting materials listed in Table 14 to prepare the corresponding target compounds.

[0566] Table 14: Compounds of Examples 244-245

[0567] Example 246: Preparation of 3-(4-amino-7-methyl-6-(perfluoroprop-2-yl)-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0568] Step 1: Synthesis of compound O-1

[0569] Into a reaction vial, was placed F-7 (100 mg, 242.5 μmol), di-tert-butyl dicarbonate (79.41 mg, 363.83 μmol), DMAP (88.9 mg, 727.7 μmol), and triethylamine (73.6 mg, 727.7 μmol), followed by 5 mL of tetrahydrofuran as solvent, the reaction was stirred at 50 °C for 2 h, after completion of the reaction, ethyl acetate was added to extract the product, the organic layer was concentrated and purified by column chromatography to give 100 mg of product O-1 in 67% yield.

[0570] Step 2: Synthesis of compound O-2

[0571] Into a reaction vial, was placed O-1 (100 mg, 163.3 μmol) and 4 mL of anhydrous tetrahydrofuran, the reaction was replaced with N2 and cooled to -78 °C, stirred for 5 min, then n-butyllithium (489.8 μmol) was added dropwise, stirred for 30 min, then N,N-diethyl-2,2,2-trifluoroacetamide (41.4 mg, 244.9 μmol,) in anhydrous tetrahydrofuran was added, and the reaction was stirred for another 30 min, after completion of the reaction, 1 mL of saturated ammonium chloride solution was added dropwise, and the reaction was slowly warmed to room temperature, ethyl acetate was added to extract the product, the organic layer was concentrated and purified by column chromatography to give 58 mg of product O-2 in 67% yield.

[0572] Step 3: Synthesis of compound O-3

[0573] Into a reaction vial, was placed O-2 (50 mg, 116.4 μmol), KF (2 mg, 34.9 μmol), TBAB (11.3 mg, 34.9 μmol) and 2 mL of anhydrous N,N-dimethylformamide, a solution of trifluoromethyltrimethylsilane (49.7 mg, 349.3 μmol) in anhydrous N,N-dimethylformamide was added dropwise under ice water bath, the reaction was stirred at room temperature for 20 h, after completion of the reaction, ethyl acetate was added to extract the product, the organic layer was concentrated and purified by column chromatography to give 21 mg of product O-3 in 34% yield.

[0574] Step 4: Synthesis of compound O-4

[0575] A reaction vial was charged with O-3 (20 mg, 40 μmol) and 2 mL of dichloromethane, DAST (38.7 mg, 240.3 μmol) was added slowly under ice water bath, the reaction was stirred for 2 h, the reaction was monitored, after the reaction was completed, water was added, dichloromethane was extracted, the water phase was extracted with ethyl acetate again, the organic layers were combined, and purified by preparative HPLC to obtain 12 mg of product O-4, with a yield of 59%.

[0576] Step 5: Preparation of the compound of Example 246

[0577] O-4 (12 mg, 23.9 μmol) was dissolved in 1 mL of dichloromethane, and boron tribromide (71.8 μmol) was added slowly under stirring, after the dropwise addition was completed, the reaction was stirred at room temperature for 3 h, after the reaction was completed, methanol was added dropwise under ice water bath to quench, sodium bicarbonate solution was added to adjust to alkaline, extracted, the water phase was extracted with ethyl acetate again, the organic layers were combined, concentrated, and purified by preparative HPLC to obtain 8.8 mg of the compound of Example 246, with a yield of 75%. 1 H NMR (400 MHz, Methanol-d4) δ 8.66 (s, 1H), 8.28 (s, 1H), 7.09 (d, J = 8.3 Hz, 1H), 6.93 (d, J = 8.2 Hz, 1H), 2.71-2.77 (m, 3H), 1.76 (s, 3H), 1.70 (s, 3H); Mass spectrum: C21H17F7N5O [M+H] + Calculated: 488.1, Found: 488.2.

[0578] Example 247: Preparation of 3-(4-amino-6-(1,1,1,3,3,3-hexafluoro-2-hydroxypropan-2-yl)-7-methyl-9H-pyrrolo[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0579] The compound of Example 247 (7.7 mg, yield: 52%) was prepared from O-3 as a starting material, according to the demethylation method of Step 5 in Example 246; 1 H NMR (400 MHz, Methanol-d4) δ 8.88 (s, 1H), 8.48 (s, 1H), 7.10 (d, J = 8.3 Hz, 1H), 6.95 (d, J = 8.3 Hz, 1H), 2.88 (s, 3H), 1.77 (s, 3H), 1.70 (s, 3H); Mass spectrum: C21H18F6N5O2 [M+H] + Calculated: 486.1, Found: 486.2.

[0580] Example 248: Preparation of 3-(4-amino-7-methyl-6-(perfluoroethyl)-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0581] Compound O-2 (50 mg, 116.44 μmol) was dissolved in 2 mL of dichloromethane, cooled to 0 °C, and DAST (18.77 mg, 116.44 μmol) was added dropwise. The reaction was stirred at room temperature for 3 h. After the reaction was completed, water was added, and dichloromethane was extracted to obtain an organic layer. Concentration and purification by preparative HPLC yielded 13 mg of the carbonyl difluorinated intermediate product. Further reference to the demethylation method in Step 5 of Example 246 was made to prepare the compound of Example 248 (8.0 mg, yield: 61%); 1 H NMR (600 MHz, chloroform-d) δ 9.18 (s, 1H), 8.46 (s, 1H), 7.11 (d, J = 8.2 Hz, 1H), 6.96 (d, J = 8.3 Hz, 1H), 2.74 (s, 3H), 1.77 (s, 3H), 1.70 (s, 3H); Mass spectrum: C20H17F5N5O [M+H] + Calculated: 438.1, Found: 438.1.

[0582] Example 249: Preparation of 3-(4-amino-7-methyl-6-(perfluoroprop-1-en-2-yl)-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0583] A reaction bottle was charged with compound O-2 (50 mg, 116.4 μmol), triphenylphosphine (61.1 mg, 232.9 μmol), and 3 mL of anhydrous N,N-dimethylformamide. The temperature was raised to 70 °C, and a solution of sodium difluorochloroacetate (35.5 mg, 232.9 μmol) in 1 mL of anhydrous N,N-dimethylformamide was added slowly with stirring. The reaction was stirred for 2 h. After the reaction was completed, water was added, and ethyl acetate was extracted to obtain an organic layer. Anhydrous sodium sulfate was added for drying, and concentration and purification by preparative HPLC yielded 6 mg of the intermediate product O-5. Further reference to the demethylation method in Step 5 of Example 246 was made to prepare the compound of Example 249 (3.5 mg, yield: 59%); 1 H NMR (400 MHz, methanol-d4) δ 8.83 (s, 1H), 8.44 (s, 1H), 7.11 (d, J = 8.3 Hz, 1H), 6.95 (d, J = 8.3 Hz, 1H), 2.59 (s, 3H), 1.77 (s, 3H), 1.71 (s, 3H); Mass spectrum: C21H17F5N5O [M+H]+ Calculated: 450.1, Found: 450.1.

[0584] Example 250: Preparation of 3-(4-amino-6-(1,1,1,3,3,3-hexafluoropropan-2-yl)-7- methyl-9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0585] Compound O-5 (10 mg, 40.82 pmol) and KF (2.37 mg, 40.82 pmol) were dissolved in 2 ml of anhydrous DMF, replaced with N2, and stirred at 80 °C for 2 h. After the reaction was completed, water was added, and ethyl acetate was extracted to obtain an organic layer. After concentration, the product was purified by preparative HPLC to obtain 4 mg of an intermediate product substituted with hexafluoroisopropyl. Further reference to the demethylation method in Example 246, Step 5, prepared Example 250 compound (1.6 mg, yield: 50%). 1 H NMR (400 MHz, Methanol-d4) δ 8.76 (s, 1H), 8.25 (s, 1H), 7.10 (d, J = 8.3 Hz, 1H), 6.93 (d, J = 8.3 Hz, 1H), 5.17-5.27 (m, 1H), 2.68 (s, 3H), 1.76 (s, 3H), 1.71 (s, 3H); Mass spectrum: C21H18F6N5O [M+H] + Calculated: 470.1, Found: 470.2.

[0586] Example 251: Preparation of 3-(4-amino-6-(2,2-difluoroethyl)-7-methyl-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0587] Step 1: Synthesis of compound O-6

[0588] Compound F-7 (269 mg, 0.653 mmol) was added to 1,4-dioxane (5 mL), (BPin)2 (199 mg, 0.784 mmol), KOAc (224 mg, 2.29 mmol), and PdCl2(dppf) (47.8 mg, 0.0653 mmol) were added, and the reaction was refluxed at 100 °C for 2 h under N2protection. After the reaction was completed, the reaction solution was cooled to room temperature, extracted with EA and water, and the organic phase was combined and washed with saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The product O-6 was separated and purified by column chromatography (PE:EA, 0→30%) to obtain 120 mg with a yield of 40%.

[0589] Step 2: Synthesis of compound O-7

[0590] Compound O-6 (120 mg, 0.261 mmol) was added into 1,4-dioxane / H2O (10:1, 2.2 mL), 2,2-difluorovinyl p-toluenesulfonate (91.7 mg, 0.392 mmol), tricyclohexylphosphine tetrafluoroborate (19 mg, 0.052 mmol), K3PO4 (194 mg, 0.913 mmol), Pd2(dba)3 (24 mg, 0.026 mmol) were added, the reaction was heated to 105 °C under N2for 2 h. After the reaction was completed, the reaction was cooled to room temperature, extracted with EA and water, the organic phase was combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, rotary evaporated, and purified by column chromatography (PE:EA, 0→30%) to give the product O-7 50 mg, yield 48%.

[0591] Step 3: Synthesis of compound O-8

[0592] Compound O-7 (50 mg, 0.126 mmol) was dissolved in MeOH (2 mL), Pd / C (10 mg, 5% w / w) was added, the reaction system was replaced with H2, and the reaction was carried out under H2for overnight. After the reaction was completed, the palladium carbon was filtered with diatomite, the filtrate was collected, and rotary evaporated to give the product O-8 22 mg, yield 44%.

[0593] Step 4: Synthesis of compound of Example 251

[0594] The compound of Example 251 was prepared according to the demethylation method of Step 5 in Reference Example 246. 1 H NMR (400 MHz, CD3OD) δ 8.45 (s, 1H), 8.13 (s, 1H), 6.98 (d, J = 8.0 Hz, 1H), 6.82 (d, J = 8.0 Hz, 1H), 5.98-6.19 (m, 1H), 3.28-3.35 (m, 2H), 2.51 (s, 3H), 1.65 (s, 3H), 1.60 (s, 3H); Mass spectrum: C 20 H 20 F2N5O [M+H] + Calculated: 384.2, Found: 384.2.

[0595] Example 252: Preparation of 3-(4-amino-7-methyl-6-(2,2,2-trifluoroethyl)-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0596] Compound O-7 (63 mg, 0.159 mmol) was dissolved in DMF (2 mL), KF (138 mg, 2.39 mmol) was added, and the reaction was reacted at 125 °C for 16 h. After the reaction was completed, the reaction solution was quenched with water, extracted with EA, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. Purification by column chromatography (DCM:MeOH = 10:1) gave 40 mg of the trifluoroethyl-substituted product. Further reference to the demethylation method in Example 246, Step 5 was used to prepare the compound of Example 252. 1 H NMR (400 MHz, CD3OD) δ 8.51 (s, 1H), 8.14 (s, 1H), 6.98 (d, J = 8.0 Hz, 1H), 6.82 (d, J = 8.0 Hz, 1H), 3.62-3.70 (m, 2H), 2.52 (s, 3H), 1.66 (s, 3H), 1.60 (s, 3H); Mass spectrum: C 20 H 19 F3N5O [M+H] + Calcd: 402.2, Found: 402.2.

[0597] Example 253: Preparation of 3-(4-amino-7-methyl-6-(thiophen-3-yl)-9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-4-chloro-2-methylphenol

[0598] Step 1: Synthesis of compound P-1

[0599] Compound F-1 (18.0 g, 71.6 mmol) was dissolved in NMP (72 mL), 2,6-dimethylpyridine (16.1 g, 150.4 mmol) was added, 6-chloro-3-methoxy-2-methylaniline (18.4 g, 107.4 mmol) was added, and the reaction was heated to 140 °C for 48 h. After the reaction was completed, the reaction solution was slowly poured into 0.5 N HCl (1.25 L) while stirring, and a gummy substance was generated at the bottom of the beaker. The filtrate was collected by filtration, and the gummy substance was dissolved in DCM. The filtrate and the gummy substance were dried over anhydrous sodium sulfate, filtered, and concentrated. Purification by column chromatography (DCM:PE, 0→30%) gave the product P-1 (20.1 g, 73% yield).

[0600] Step 2: Synthesis of compound P-2

[0601] Compound P-2 (12.0 g, 32.0 mmol) was dissolved in THF (120 mL), di-tert-butyl dicarbonate (31.5 g, 144 mmol) was added, DMAP (3.98 g, 32.0 mmol) was added slowly, the temperature was raised to 60 °C and stirred for 16 h. After the reaction was completed, it was cooled to room temperature, quenched with water, extracted with EA, the organic phase was washed with water, saturated brine, the organic phase was dried over anhydrous sodium sulfate, filtered, and rotary evaporated. Purification was performed by column chromatography (PE:EA, 0→25%) to give the product P-3 (6.0 g, 40% yield).

[0602] Step 3: Synthesis of compound P-3

[0603] Compound P-2 (12.0 g, 32.0 mmol) was dissolved in THF (120 mL), di-tert-butyl dicarbonate (31.5 g, 144 mmol) was added, DMAP (3.98 g, 32.0 mmol) was added slowly, the temperature was raised to 60 °C and stirred for 16 h. After the reaction was completed, it was cooled to room temperature, quenched with water, extracted with EA, the organic phase was washed with water, saturated brine, the organic phase was dried over anhydrous sodium sulfate, filtered, and rotary evaporated. Purification was performed by column chromatography (PE:EA, 0→25%) to give the product P-3 (6.0 g, 40% yield).

[0604] Step 4: Synthesis of compound P-4

[0605] Compound P-3 (6.0 g, 12.7 mmol) was first dissolved in EA (20 mL), then EtOH (60 mL) was added, Fe (4.27 g, 76.2 mmol) was added, a solution of ammonium chloride (0.85 g, 15.9 mmol) and water (3 mL) was added, the temperature was raised to 80 °C, and stirred overnight. After the reaction was completed, the iron powder was filtered with diatomite, the filtrate was collected, rotary evaporated, and purified by column chromatography (PE:EA, 0→40%) to give the product P-4 (3.60 g, 64% yield).

[0606] Step 5: Synthesis of compound P-5

[0607] Compound P-4 (3.6 g, 8.15 mmol) was dissolved in super dry MeCN (40 mL), CuBr2(1.82 g, 8.15 mmol) was added, the air was replaced with N2, tert-butyl nitrite (0.92 g, 8.97 mmol) was added under N2protection, the temperature was raised to 60 °C, and reacted for 1 h. After the reaction was completed, it was rotary evaporated, and purified by column chromatography (PE:EA, 0→25%) to give the product P-5 (1.70 g, 41% yield).

[0608] Step 6: Synthesis of compound P-6

[0609] Compound P-5 (1.70 g, 3.36 mmol) was weighed into a reaction flask, formamide (20 mL) was added, and the temperature was raised to 205 °C for 40 min. The reaction was quenched with water, extracted with EA, and the organic phase was washed with water, saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. Compound P-6 (900 mg, 65% yield) was obtained by column chromatography (PE:EA, 0→35%).

[0610] Step 7: Synthesis of compound P-7

[0611] Compound P-6 (900 mg, 2.18 mmol) was dissolved in DCM (10 mL), and BBr3 (10.9 mL, 10.9 mmol, 1 min DCM) was added. The reaction was stirred at room temperature for 1.5 h. After the reaction was completed, the reaction solution was added dropwise to 100 mL of saturated aqueous sodium bicarbonate solution, stirred for 15 min, and filtered to obtain a solid. The solid was washed with water, and the aqueous phase was extracted once with DCM. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The obtained solid was dissolved in MeOH, and the filtrate was combined and concentrated. Compound P-7 (600 mg, 66% yield) was obtained by purification with DCM slurry.

[0612] Step 8: Synthesis of compound of Example 253

[0613] Compound P-7 (40.0 mg, 0.0955 mmol) and thiophene-2-boronic acid pinacol ester (26.1 mg, 0.124 mmol) were dissolved in dioxane (1.5 mL) and water (0.15 mL), and potassium carbonate (59.4 mg, 0.430 mmol) was added. Pd(dppf)Cl2 (7.35 mg, 0.01 mmol) was added. The reaction was carried out at 100 °C under N2for 3 h. After the reaction was completed, it was concentrated and purified by plate chromatography (DCM / MeOH=10:1) to obtain compound of Example 253 (20.0 mg, 50% yield). 1 H NMR (400 MHz, MeOH-d4) δ 8.64 (s, 1H), 8.25 (s, 1H), 7.52 (d, J = 4.0 Hz, 1H), 7.37 (d, J = 8.0 Hz, 1H), 7.15-7.19 (m, 2H), 2.59 (s, 3H), 1.87 (s, 3H); Mass spectrum: C 21 H 17 ClN5OS [M+H] + Calculated: 422.0, Found: 422.0.

[0614] Example 254: Preparation of 3-(4-amino-7-methyl-6-(prop-1-en-2-yl)-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-4-chloro-2-methylphenol

[0615] Compound P-7 (150.0 mg, 0.358 mmol) and allylboronic acid pinacol ester (78.2 mg, 0.467 mmol) were dissolved in dioxane (4 mL) and water (0.4 mL), potassium carbonate (222 mg, 1.61 mmol) was added, Pd(dppf)Cl2(26.1 mg, 0.0358 mmol) was added. The reaction was reacted at 100 degree N2environment for 3 hours, after the reaction was completed, concentrated, separated and purified by plate chromatography (DCM / MeOH = 10:1) to obtain the compound of Example 254 (50.0 mg, yield 37%). 1 H NMR (400 MHz, MeOH-d4) δ 8.42 (s, 1H), 8.23 (s, 1H), 7.36 (d, J = 8.0 Hz, 1H), 6.86 (d, J = 8.0 Hz, 1H), 5.37 (s, 1H), 5.01 (s, 1H), 2.55 (s, 3H), 2.17 (s, 3H), 1.85 (s, 3H); Mass spectrum: C 20 H 19 ClN5O [M+H] + Calculated: 380.1, Found: 380.0.

[0616] Example 255: Preparation of 3-(4-amino-6-isopropyl-7-methyl-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-4-chloro-2-methylphenol

[0617] The compound of Example 254 (40 mg, 0.105 mmol) was dissolved in methanol (2 mL), 10% wet Pd / C (4 mg) was added, the reaction was reacted at room temperature for 15 hours under hydrogen atmosphere, after the reaction was completed, filtered, concentrated, separated and purified by HPLC preparation to obtain the compound of Example 255 (11.0 mg, yield 27%). 1 H NMR (400 MHz, MeOH-d4) δ 8.45 (s, 1H), 8.11 (s, 1H), 7.26 (d, J = 7.9 Hz, 1H), 6.75 (d, J = 7.9 Hz, 1H), 3.24-3.27 (m, 1H), 2.50 (s, 3H), 1.74 (s, 3H), 1.29 (t, J = 7.5 Hz, 6H); Mass spectrum: C 20 H 21CIN5O [M+H] + Calculated: 382.1, Found: 382.1.

[0618] Example 256: Preparation of 3-(4-amino-6-isopropyl-7-methyl-9H-pyrrolo[3',2':4,5]pyrrolo[2,3- d]pyrimidin-9-yl)-4-chloro-2-methylphenol

[0619] Step 1: Synthesis of compound P-8

[0620] Compound P-2 (5.95 g, 16 mmol) was dissolved in THF (65 mL), di-tert-butyl dicarbonate (15.75 g, 68 mmol) was added, DMAP (1.96 g, 16 mmol) was added slowly, the temperature was raised to 60 °C, and stirred at 60 °C for 20 min. After the reaction was completed, it was cooled to room temperature, quenched with water, extracted with EA, the organic phase was washed with water, saturated brine, and dried over anhydrous sodium sulfate. After filtration, it was concentrated, and the product P-8 (1.3 g, yield 14%) was obtained by column chromatography (PE:EA, 0→100%) separation and purification.

[0621] Step 2: Synthesis of compound P-9

[0622] Compound P-9 was prepared according to the synthesis method of reference compound P-4.

[0623] Step 3: Synthesis of compound P-10

[0624] Compound P-9 (0.34 g, 0.63 mmol) was dissolved in DMF (7 mL), 1,4-diiodobutane (0.292 g, 0.94 mmol) was added first, and then K2CO3 (0.565 g, 4.1 mmol) was added. The temperature was raised to 75 °C, and stirred for 2 h. Then 1,4-diiodobutane (0.195 g, 0.63 mmol) and K2CO3 (0.261 g, 1.89 mmol) were added, and the reaction was continued at 75 °C for 2 h. After cooling to room temperature, it was quenched with water, and the aqueous phase was extracted with EA. The organic phase was washed with saturated brine once, dried over anhydrous sodium sulfate, filtered, and concentrated. The sample was mixed, and the product P-10 (0.13 g, yield 35%) was obtained by column chromatography (PE:EA, 0→100%) separation and purification.

[0625] Step 4-5: Preparation of the compound of Example 256

[0626] The compound of Example 256 was prepared according to the method of Example 253, steps 6-7. 1H NMR (400 MHz, Methanol-d4) δ 8.20 (s, 1H), 8.09 (s, 1H), 7.25 (d, J = 8.0 Hz, 1H), 6.73 (d, J = 8.0 Hz, 1H), 3.11-3.14 (m, 4H), 2.45 (s, 3H), 1.89-1.96 (m, 4H), 1.73 (s, 3H); Mass spectrum: C 21 H 22 ClN6O [M+H] + Calculated: 409.1, Found: 409.0.

[0627] Examples 257-265:

[0628] The synthesis method of Reference Examples 253-256 was used to prepare the corresponding target compounds using the starting materials listed in Table 15.

[0629] Table 15: Compounds of Examples 257-265

[0630] Example 266: Preparation of 4-(4-amino-7-methyl-6-(thiophen-2-yl)-9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-3,5-dimethylbenzene-1,2-diol

[0631] The compound of Example 157 (40 mg, 99.63 μmol) was weighed into CHCl3(3 mL) and MeOH (2 mL), IBX (33.48 mg, 119.56 μmol) was added at 0 °C, and the reaction was continued at 0 °C for 3 h. LCMS monitoring showed that the starting material was consumed and the product was formed. Water and EA were added for extraction, and the organic layer was washed with brine. The solvent was removed under vacuum, and the product of Example 266 (3 mg, 7.0% yield) was obtained by high performance liquid chromatography preparation. 1 H NMR (400 MHz, DMSO-d6) δ 9.51 (s, 1H), 8.92 (s, 1H), 8.36 (s, 1H), 8.06 (s, 2H), 7.70 (dd, J = 5.1, 1.2 Hz, 1H), 7.33 (dd, J = 3.5, 1.2 Hz, 1H), 7.22 (dd, J = 5.2, 3.5 Hz, 1H), 6.68 (s, 1H), 2.56 (s, 3H), 1.66 (s, 3H), 1.60 (s, 3H); Mass spectrum: C22H20N5O2S [M+H] + Calculated: 418.1, Found: 418.1.

[0632] Example 267: Preparation of (R)-3-(4-amino-7-methyl-6-(thiophen-2-yl)-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0633] The chiral isomers of the compound of Example 157 (6.0 g) were separated by SFC method using preparative equipment Shimadzu LC-20AD XR and chiral column CHIRALCEL OZ-H (OZH0CD-TC005, 0.46 cm x 15 cm), eluent: CO2 / MeOH / DEA = 70 / 30 / 0.1 (V / V / V); flow rate: 2 ml / min; temperature: 35 °C. Component 1 (t R = 4.74 min, 2.36 g, >98% ee).[a] D 22 = 33.1 (c = 1, MeOH).

[0634] Example 268: (S)-3-(4-amino-7-methyl-6-(thiophen-2-yl)-9H-pyrido[3',2':4,5]pyrrolo[2,3- d]pyrimidin-9-yl)-2,4-dimethylphenol

[0635] The chiral isomers of the compound of Example 157 (6.0 g) were separated by SFC method using preparative equipment Shimadzu LC-20AD XR and chiral column CHIRALCEL OZ-H (OZH0CD-TC005, 0.46 cm x 15 cm), eluent: CO2 / MeOH / DEA = 70 / 30 / 0.1 (V / V / V); flow rate: 2 ml / min; temperature: 35 °C. Component 2 (t R = 5.71 min, 2.58 g, >98% ee).[a] D 22 = -28.3 (c = 1, MeOH).

[0636] Example 269: Preparation of (R)-3-(4-amino-7-methyl-6-(pyrrolidin-1-yl)-9H-pyrido[3',2':4,5] pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0637] The chiral isomers of the compound of Example 152 (6.0 g) were separated by SFC method using preparative equipment Shimadzu LC-20AD and chiral column CHIRALCEL OZ-H (OZH0CD-TC005, 0.46 cm x 15 cm), eluent: CO2 / MeOH / DEA = 70 / 30 / 0.1 (V / V / V); flow rate: 1 ml / min; temperature: 35 °C. Component 1 (t R = 4.18 min, 2.47 g, >97% ee).[a] D 22 = 19.0 (c = 1, MeOH).

[0638] Example 270: Preparation of (S)-3-(4-amino-7-methyl-6-(pyrrolidin-1-yl)- 9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0639] The chiral isomers of the compound of Example 152 (6.0 g) were separated by SFC method using preparative equipment Shimadzu LC-20AD and chiral column CHIRALCEL OZ-H (OZH0CD-TC005, 0.46 cm x 15 cm), eluent: CO2 / MeOH / DEA = 70 / 30 / 0.1 (V / V / V); flow rate: 1 ml / min; temperature: 35 °C. Component 2 (t R = 5.15 min, 2.18 g, >97% ee).[a] D 22 = -22.2 (c = 1, MeOH).

[0640] Example 271: Preparation of 3-(4-amino-7-methyl-6-octyl-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0641] Step 1: Synthesis of compound Q-2

[0642] A microwave reaction tube was charged with F-7 (30 mg, 72.77 μmol), Q-1 (12.03 mg, 109.15 μmol), dichlorobis(triphenylphosphine)palladium (5.11 mg, 7.28 μmol), CuI (692 μg, 3.64 μmol), triethylamine (58.91 mg, 582.13 μmol) and anhydrous DMF as solvent, replaced with N2, heated to 120 °C for 3 h. After the reaction was completed, EA was used for extraction, and the organic layer was concentrated to give 10 mg of product Q-2, with a yield of 31%.

[0643] Step 2: Synthesis of compound Q-3

[0644] Into a reaction vial was placed Q-2 (10 mg, 22.65 μmol), Pd / C (5% Pd / C, wet with ca. 55% water) (2.41 mg, 22.65 μmol), MeOH (5 mL), replaced with N2, stirred at room temperature overnight. The reaction was complete. The reaction mixture was filtered to give a filtrate, which was concentrated to give 8 mg of product Q-3 in 79% yield.

[0645] Step 3: Preparation of compound of Example 271

[0646] Into a reaction vial was placed Q-3 (8 mg, 17.95 μmol), DCM (2 mL) to dissolve, BBr3 (2 M in DCM) (107.72 μmol), stirred for 2 h. The reaction was complete. Methanol was added to quench, NaHCO3 solution was added to adjust to basic, the aqueous phase was extracted with EA, the organic layers were combined and concentrated to give 5.2 mg of compound of Example 271 in 66% yield. 1 H NMR (600 MHz, Methanol-d4) δ 8.47 (s, 1H), 8.22 (s, 1H), 7.11 (d, J = 8.3 Hz, 1H), 6.94 (d, J = 8.3 Hz, 1H), 2.83-2.87 (m, 2H), 2.58 (s, 3H), 1.71-1.77 (m, 8H), 1.48 -1.51 (m, 2H), 1.37-1.44 (m, 7.2 Hz, 2H), 1.30-1.35 (m, 6H), 0.92 (t, J = 6.9 Hz, 3H); Mass spectrum: C 26 H 34 N5O [M+H] + Calculated: 432.3, Found: 432.3.

[0647] Example 272: Preparation of 3-(4-amino-6-(8-bromooctyl)-7-methyl-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0648] The compound of Example 272 was synthesized according to the procedures of Reference Example 271, Steps 1-3. 6.3 mg of compound of Example 272 was prepared in 56% yield. 1H NMR (600 MHz, Methanol-d4) δ 8.47 (s, 1H), 8.22 (s, 1H), 7.10 (d, J = 8.3 Hz, 1H), 6.94 (d, J = 8.3 Hz, 1H), 2.83-2.88 (m, 2H), 2.58 (s, 3H), 1.82-1.90 (m, 2H), 1.74 (d, J = 29.9 Hz, 8H), 1.30-1.54 (m, 10H); Mass: C 26 H 32 BrN5O [M+H] + Calculated: 510.2, Found: 510.2.

[0649] Example 273: Preparation of 3-(4-amino-6-(8-hydroxyoctyl)-7-methyl-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0650] Step 1: Synthesis of compounds Q-6 and Q-7

[0651] Into a reaction vial was placed Q-5 (500 mg, 3.96 mmol), anhydrous DMSO (2 mL) as solvent, KOH (266.78 mg, 4.75 mmol), oxirane (261.81 mg, 5.94 mmol), replaced with N2, reacted at room temperature for 12 h, TLC and TLCMS monitoring, the reaction was completed, EA extraction, organic layer was obtained, concentrated, column chromatography (EA: PE = 0-15%-30%-50%-80%) to obtain target compounds Q-6 (52 mg, yield 7%) and Q-7 (30 mg, yield 3%).

[0652] Step 2: Synthesis of compound Q-8

[0653] Into a reaction vial was placed F-8 (30 mg, 75.33 μmol), Q-5 (14.26 mg, 112.99 μmol), bis(triphenylphosphine)palladium dichloride (10.57 mg, 15.07 μmol), CuI (1.43 mg, 7.53 μmol), triethylamine (60.98 mg, 602.63 μmol), anhydrous DMF as solvent, replaced with N2, heated to 120 °C for 3 h, the reaction was completed, EA extraction, organic layer was obtained, concentrated, 15 mg of target compound Q-8 was prepared, yield 44%.

[0654] Step 3: Preparation of the compound of Example 273

[0655] To a flask was added O-2 (296 mg, 0.56 mmol), di-tert-butyl dicarbonate (183.81 mg, 0.84 mmol), DMAP (205.78 mg, 1.68 mmol), triethylamine (170.44 mg, 1.68 mmol) in THF as solvent, the reaction was stirred at 50 °C for 2 h, the reaction was completed, extracted with EA, column chromatography, 320 mg of compound R-1 was obtained with a yield of 91 %. 1 H NMR (600 MHz, Methanol-d4) δ 8.56 (s, 1H), 8.33 (s, 1H), 7.00 (d, J = 8.3 Hz, 1H), 6.85 (d, J = 8.3 Hz, 1H), 3.44 (t, J = 6.6 Hz, 2H), 2.74 - 2.79 (m, 2H), 2.51 (s, 3H), 1.63 - 1.67 (m, 5H), 1.59 (s, 3H), 1.63 - 1.67 (m, 4H), 1.27 - 1.35 (m, 6H); Mass spectrum: C 26 H 34 N5O2[M+H] + Calculated: 448.3, Found: 448.3.

[0656] Examples 274-275 of Table 16 were prepared according to the synthetic method of Example 273.

[0657] Table 16: Compounds of Examples 274-275

[0658] Example 276: Preparation of 3-(4-amino-7-methyl-6-(1,1,1,2-tetrafluoropropan-2-yl-3,3,3-d3)-9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0659] Step 1: Synthesis of compound R-1

[0660] To a flask was added O-2 (296 mg, 0.56 mmol), di-tert-butyl dicarbonate (183.81 mg, 0.84 mmol), DMAP (205.78 mg, 1.68 mmol), triethylamine (170.44 mg, 1.68 mmol) in THF as solvent, the reaction was stirred at 50 °C for 2 h, the reaction was completed, extracted with EA, column chromatography, 320 mg of compound R-1 was obtained with a yield of 91 %.

[0661] Step 2: Synthesis of compound R-2

[0662] Into a reaction vial, was placed R-1 (320 mg, 508.24 µmol), anhydrous THF was added as solvent, N2 protection, ice water bath stirring for 5 min, slowly droped deuterium (d3) methyl magnesium iodide (111.83 mg, 660.71 µmol), continue to stir for 15 min, the reaction was completed, added aqueous solution of citric acid, extracted with EA, obtained organic layer, column chromatography, obtained 50 mg of compound R-2, the yield was 15%.

[0663] Step 3: Synthesis of compound R-3

[0664] Into a reaction vial, was placed R-2 (20 mg, 30.83 µmol), dissolved in DCM, slowly added DAST (29.82 mg, 184.99 µmol) under ice water bath, stirred for 2 h, monitored the reaction, the reaction was completed, added water, extracted with DCM, the aqueous phase was extracted with EA again, combined the organic layers, prepared 4.2 mg of target compound R-3, the yield was 24%.

[0665] Step 4: Preparation of compound of Example 276

[0666] Into a reaction vial, was placed R-3 (5 mg, 7.68 µmol), dissolved in DCM (2 mL), droped BBr3 (46.11 µmol), stirred at 40 ℃ for 4 h, monitored the reaction, the reaction was completed, added methanol to quench, added NaHCO3 solution to adjust to alkaline, extracted with DCM, the aqueous phase was extracted with EA again, combined the organic layers, concentrated, prepared, obtained 1.5 mg of compound of Example 276, the yield was 43%. 1 H NMR (600 MHz, Methanol-d4) δ 8.88 (s, 1H), 8.42 (s, 1H), 7.13 (dd, J = 8.4, 3.1 Hz, 1H), 6.97 (d, J = 8.3 Hz, 1H), 2.77 (d, J = 6.2 Hz, 3H), 1.79 (d, J = 15.5 Hz, 3H), 1.73 (d, J = 16.1 Hz, 3H); Mass spectrum: C21H17D3F4N5O [M+H] + Calculated: 437.2, Found: 437.2.

[0667] Preparation of Example 277: 3-(4-amino-7-methyl-6-(1,1,1-trifluoro-2-hydroxypropan-2-yl-3,3,3-d3)-9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol and Example 278: 3-(4-amino-7-methyl-6-(1,1,1-trifluoro-2-hydroxypropan-2-yl-3,3,3-d3)-9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0668] To the reaction bottle, R-1 (10 mg, 15.42 pmol) was added, then dissolved in DCM (2 mL), BBr3 (92.49 pmol) was added dropwise, and the reaction was stirred at 40 °C for 4 h. The reaction was monitored, and after the reaction was completed, methanol was added for quenching, NaHCO3 solution was added to adjust to alkaline, DCM was added for extraction, and the water phase was extracted with EA. The organic layers were combined and concentrated to prepare two groups of diastereoisomers, 1.4 mg of the compound of Example 277, with a yield of 20%, 1 H NMR (600 MHz, Methanol-d4) d 8.96 (s, 1H), 8.48 (s, 1H), 7.13 (d, J = 8.3 Hz, 1H), 6.98 (d, J = 8.3 Hz, 1H), 2.89 (s, 3H), 1.80 (s, 3H), 1.70 (s, 3H); Mass spectrum: C 21 H 17 D3F3N5O2 [M+H] + Calculated: 435.2, Found: 435.2. Preparation obtained 1.6 mg of the compound of Example 278, with a yield of 22%, 1 H NMR (600 MHz, Methanol-d4) d 8.96 (s, 1H), 8.48 (s, 1H), 7.13 (d, J = 8.3 Hz, 1H), 6.98 (d, J = 8.3 Hz, 1H), 2.89 (s, 3H), 1.80 (s, 3H), 1.70 (s, 3H); Mass spectrum: C 21 H 17 D3F3N5O2 [M+H] + Calculated: 435.2, Found: 435.2.

[0669] Preparation of Example 279: 3-(4-amino-7-methyl-6-(1,1,1-trifluoropropan-2-yl-3,3,3-d3)-9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol and Example 280: 3-(4-amino-7-methyl-6-(1,1,1-trifluoropropan-2-yl-3,3,3-d3)-9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0670] Step 1: Synthesis of compound R-4

[0671] A reaction vial was charged with R-2 (80 mg, 123.33 μmol), dissolved in pyridine (1.95 mL), and added with dichlorosulfoxide (88.03 mg, 739.96 μmol). The reaction was stirred at room temperature overnight. Upon completion, the excess pyridine and dichlorosulfoxide were removed by concentration. The reaction was quenched with saturated NaHC03solution. The organic layer was extracted with DCM and concentrated. Purification gave 23 mg of compound R-4 in 20% yield and 7 mg of compound R-5 in 20% yield.

[0672] Step 2: Synthesis of compound R-6

[0673] A reaction vial was charged with R-4 (28 mg, 49.26 μmol), Pt02(11.19 mg, 49.26 μmol), methanol (2 mL), and ethyl acetate (2 mL) to replace hydrogen gas. The reaction was stirred at room temperature overnight. Upon completion, the reaction was filtered, concentrated, and dried in vacuum. The crude compound R-6 was used directly in the next step.

[0674] Step 3: Preparation of Example 279 compound and Example 280 compound

[0675] A reaction vial was charged with R-6 (20 mg, 37.55 μmol), dissolved in DCM, and added with BBr3(225.32 μmol). The reaction was stirred at room temperature for 4 h. Upon completion, the reaction was quenched with methanol. The pH was adjusted to basic with saturated NaHC03solution. The organic layer was extracted with DCM and the aqueous phase was extracted with EA. The combined organic layers were concentrated. Two diastereomers were prepared, 4.9 mg of Example 279 compound in 30% yield, 1 H NMR (400 MHz, Methanol-d4) δ 8.96 (s, 1H), 8.46 (s, 1H), 7.12 (d, J = 8.3 Hz, 1H), 6.97 (d, J = 8.3 Hz, 1H), 4.15 (q, J = 9.5, 9.0 Hz, 1H), 2.70 (s, 3H), 1.77 (s, 3H), 1.73 (s, 3H); Mass spectrum: C 21 H 18 D3F3N5O [M+H] + Calculated: 419.2, Found: 419.4. Preparation gave 6.7 mg of Example 280 compound in 42% yield, 1 H NMR (400 MHz, Methanol-d4) δ 8.94 (s, 1H), 8.43 (s, 1H), 7.13 (d, J = 8.3 Hz, 1H), 6.97 (d, J = 8.3 Hz, 1H), 4.16 (t, J = 9.8 Hz, 1H), 2.69 (s, 3H), 1.79 (s, 3H), 1.71 (s, 3H); Mass spectrum: C 21 H 18D3F3N5O [M+H] + Calculated: 419.2, Found: 419.4.

[0676] Examples 281-282 of Table 17 were prepared according to the synthetic method of Examples 279-280.

[0677] Table 17: Compounds of Examples 281-282

[0678] Examples 281-282 of Table 17 were prepared according to the synthetic method of Examples 279-280.

[0679] Step 1: Synthesis of compound R-7

[0680] Into a reaction vial was placed R-5 (14 mg, 24.69 μmol), then deuterated (d4) methanol (2 mL) was added to dissolve, replaced with deuterium gas, stirred at room temperature overnight, after the reaction was completed, filtered, concentrated, the crude compound R-7 was used directly for the next step.

[0681] Step 2: Preparation of Example 283 compound and Example 284 compound

[0682] Into a reaction vial was placed R-7 (10 mg, 18.74 μmol), then DCM was added to dissolve, then BBr3 (112.45 μmol) was added, stirred at room temperature for 4 h, after the reaction was completed, methanol was added to quench, then saturated NaHCO3 was added to adjust the pH to basic, extracted with DCM, the aqueous phase was extracted with EA again, the combined organic layers were concentrated, two groups of diastereoisomers were prepared, 2.0 mg of Example 283 compound, yield was 24%, 1 H NMR (600 MHz, Methanol-d4) δ 8.76 (s, 1H), 8.25 (s, 1H), 7.11 (d, J = 8.3 Hz, 1H), 6.95 (d, J = 8.3 Hz, 1H), 2.66 (s, 3H), 1.76 (s, 3H), 1.74 (s, 3H); Mass spectrum: C 21 H 17 D4F3N5O [M+H] +Calculated: 420.2, found: 420.2. Preparation of 3.8 mg of the compound of Example 284 with a yield of 47%, was obtained, 1 HNMR (400 MHz, Methanol-d4) δ 8.88 (s, 1H), 8.37 (s, 1H), 7.01 (d, J = 8.3 Hz, 1H), 6.86 (d, J = 8.3 Hz, 1H), 2.58 (s, 3H), 1.67 (s, 3H), 1.59 (s, 3H); Mass spectrum: C 21 H 17 D4F3N5O [M+H] + Calculated: 420.2, found: 420.2.

[0683] Example 285: Preparation of 3-(4-amino-7-methyl-6-(1,1,1,2-tetrafluoropropan-2-yl-3,3,3-d3)-9H-pyrrolo[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0684] Step 1: Synthesis of compound R-8

[0685] A reaction bottle was added with F-7 (600 mg, 1.46 mmol), dimethyl oxalate (1.72 g, 14.55 mmol), palladium dichloride bis(triphenylphosphine) (102.15 mg, 145.53 μmol), DMAP (1.78 g, 14.55 mmol), MeOH (10 mL) was added as a solvent, N2 was replaced, and the reaction was stirred at 140 °C (pressure 14 bar) for 2 h after microwave heating. After the reaction was completed, it was concentrated, and the crude product was subjected to flash column chromatography. The crude product was dissolved in DCM (10 mL), and di-tert-butyl dicarbonate (317.62 mg, 1.46 mmol), triethylamine (736.32 mg, 7.28 mmol), and DMAP (35 mg, 0.29 mmol) were added. The reaction was stirred at 40 °C overnight after heating. After the reaction was completed, it was concentrated and subjected to column chromatography to obtain 700 mg of compound R-8 with a yield of 81%.

[0686] Step 2: Synthesis of compound R-9

[0687] A reaction bottle was added with R-8 (200 mg, 338.04 μmol), and dry THF (2 mL) was added for dissolution. N2 was replaced, and deuterated methyl (d3) magnesium iodide (171.65 mg, 1.01 mmol) was added under ice water bath. The reaction was heated to 60 °C for 4 h. After the reaction was completed, it was cooled to room temperature, and saturated NH4Cl solution was added. Extraction was performed with EA to obtain an organic layer, which was concentrated and subjected to column chromatography to obtain 150 mg of the target compound R-9 with a yield of 74%.

[0688] Step 3: Preparation of the compound of Example 285

[0689] To a reaction vial was added the compound of Example 285 (10 mg, 26.08 μmol), DCM (2 mL) to dissolve, then DAST (12.61 mg, 78.23 μmol) was added dropwise. The reaction was stirred overnight. After the reaction was completed, saturated NaHC03solution was added, and the mixture was extracted with DCM. The aqueous phase was extracted with EA again. The organic phases were combined and concentrated to give 8.7 mg of the compound of Example 286 in 85% yield. 1 H NMR (400 MHz, Methanol-d4) δ 8.66 (s, 1H), 8.23 (s, 1H), 8.15 (s, 1H), 7.11 (d, J = 8.3 Hz, 1H), 6.94 (d, J = 8.3 Hz, 1H), 2.85 (s, 3H), 1.77 (s, 3H), 1.71 (s, 3H); Mass spectrum: C 21 H 18 D6N5O2 [M+H] + Calculated: 384.2, Found: 384.3.

[0690] Example 286: Preparation of 3-(4-amino-6-(2-fluoroprop-2-yl-1,1,1,3,3,3-d6)-7-methyl-9H-pyrrolo[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0691] To a reaction vial was added the compound of Example 285 (10 mg, 26.08 μmol), DCM (2 mL) to dissolve, then DAST (12.61 mg, 78.23 μmol) was added dropwise. The reaction was stirred overnight. After the reaction was completed, saturated NaHC03solution was added, and the mixture was extracted with DCM. The aqueous phase was extracted with EA again. The organic phases were combined and concentrated to give 8.7 mg of the compound of Example 286 in 85% yield. 1 H NMR (600 MHz, Methanol-d4) δ 8.89-8.85 (m, 1H), 8.48 (d, J = 8.5 Hz, 1H), 7.12 (d, J = 8.3 Hz, 1H), 6.97 (d, J = 8.4 Hz, 1H), 2.89 (s, 1H), 2.78 (d, J = 3.2 Hz, 2H), 1.78 (d, J = 4.4 Hz, 3H), 1.71 (d, J = 4.9 Hz, 3H); Mass spectrum: C 21 H 17 D6FN5O [M+H] + Calculated: 386.2, Found: 386.2.

[0692] Example 287: Preparation of 3-(4-amino-7-methyl-6-prop-2-yl-1,1,1,3,3,3-d6)-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0693] Step 1: Synthesis of compound R-10

[0694] Into a reaction vial was placed R-9 (110 mg, 184.03 μmol), DCM (2 mL) and TFA (1 mL). The reaction was stirred at room temperature for 3-4 h. Upon completion, the DCM and excess TFA were removed by concentration. Saturated NaHC03solution was added, and the mixture was extracted with EA. The organic layer was dried over anhydrous Na2S04and concentrated to give the crude compound R-10, which was used directly in the next step.

[0695] Step 2: Synthesis of compound R-11

[0696] Into a reaction vial was placed R-10 (25 mg, 62.89 μmol), TFA (1 mL), and triethylsilane (36.56 mg, 314.46 μmol). The reaction was stirred at 80 °C for 2 h. Upon completion, the TFA was removed by concentration. Saturated NaHC03solution was added, and the mixture was extracted with EA. The organic layer was concentrated to give 6 mg of compound R-11 in 25% yield and 13 mg of compound R-12 in 54% yield.

[0697] Step 3: Preparation of the compound of Example 287

[0698] Into a reaction vial was placed R-11 (6 mg, 15.73 μmol), DCM (2 mL), and BBr3(94.36 μmol). The reaction was stirred at room temperature for 4 h. Upon completion, the reaction was quenched with methanol. Saturated NaHC03solution was added to make the mixture basic. The mixture was extracted with DCM and EA. The organic layer was concentrated and purified by HPLC to give 2.8 mg of the compound of Example 287 in 45% yield. 1 H NMR (600 MHz, Methanol-d4) δ 8.46 (s, 1H), 8.10 (s, 1H), 6.99 (d, J = 8.3 Hz, 1H), 6.82 (d, J = 8.3 Hz, 1H), 3.20 (s, 1H), 2.50 (s, 3H), 1.65 (s, 3H), 1.60 (s, 3H); Mass spectrum: C 21 H 18 D6N5O [M+H] + Calculated: 368.2, Found: 368.3.

[0699] Example 288 of Table 18 was prepared according to the synthetic method of Example 287.

[0700] Table 18: Compounds of Example 288

[0701] Example 289: Preparation of 3-(4-amino-7-methyl-6-(propan-2-yl-d7)-9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl-2-d)-2,4-dimethylphenol

[0702] Step 1: Synthesis of compound R-13

[0703] Into a reaction vial was placed R-12 (40 mg, 105.69 µmol), followed by PtO2 (12.00 mg, 52.84 µmol), deuterated methanol (1.5 mL) was added as solvent, replaced with N2 for three times, replaced with D2, heated to 50 °C and stirred overnight. The reaction was complete, filtered, concentrated to give crude compound R-13, which was used directly in the next step.

[0704] Step 2: Preparation of compound of Example 289

[0705] Into a reaction vial was placed R-13 (10.00 mg, 26.07 µmol), followed by DCM (2 mL) to dissolve, BBr3 (156.45 µmol) was added slowly, stirred at room temperature for 4 h. The reaction was complete, methanol was added to quench, saturated NaHCO3 solution was added to adjust to basic, extracted with DCM, the aqueous phase was extracted with EA again, combined organic layers, concentrated to give 6.7 mg of compound of Example 289 in 69% yield. 1 H NMR (400 MHz, Methanol-d4) δ 8.59 (s, 1H), 7.11 (d, J = 8.3 Hz, 1H), 6.94 (d, J = 8.3 Hz, 1H), 2.61 (s, 3H), 1.77 (s, 3H), 1.72 (s, 3H); Mass spectrum: C 21 H 16 D8N5O [M+H] + Calculated: 370.3, Found: 370.3.

[0706] Examples 290-292 of Table 19 were prepared according to the synthetic method of Example 289.

[0707] Table 19: Compounds of Examples 290-292

[0708] Example 293: Preparation of 3-(4-amino-7-methyl-6-(propan-2-yl-d7)-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)-2,4-dimethylphenol

[0709] Step 1: Synthesis of compound R-14

[0710] Into a reaction vial, was placed R-12 (50 mg, 132.10 μmol), di-tert-butyl dicarbonate (43.24 mg, 198.15 μmol), DMAP (48.4 mg, 396.31 μmol), triethylamine (40.10 mg, 396.31 μmol), THF as solvent, the reaction was stirred at 50 °C for 2 h, after completion of the reaction, extracted by EA, column chromatography, 62 mg compound R-14 was obtained with a yield of 81 %.

[0711] Step 2: Synthesis of compound R-15

[0712] Into a reaction vial, was placed R-14 (62 mg, 107.13 μmol), then PtO2 (7.30 mg, 32.14 μmol) was added, MeOD (1.5 mL) was added as solvent, replaced with N2 for 3 times, and then replaced with D2, stirred at room temperature for 4 h, after completion of the reaction, filtered, concentrated, the crude product was dissolved in DCM, then di-tert-butyl dicarbonate (43.24 mg, 198.15 μmol), DMAP (48.4 mg, 396.31 μmol), triethylamine (40.10 mg, 396.31 μmol) was added, stirred at 40 °C for 3 h, TLC monitoring, after completion of the reaction, column chromatography, 46 mg target compound R-15 was obtained with a yield of 73 %.

[0713] Step 3: Preparation of the compound of Example 293

[0714] Into a reaction vial, was placed R-15 (46 mg, 78.94 μmol), then DCM (3 mL) was added to dissolve, BBr3 (473.62 μmol) was added slowly, stirred at room temperature for 4 h, after completion of the reaction, methanol was added to quench, saturated NaHCO3 solution was added to adjust to alkaline, extracted by DCM, the aqueous phase was extracted by EA again, the organic phases were combined, concentrated, 19.6 mg compound of Example 293 was obtained with a yield of 66 %. 1 H NMR (600 MHz, Methanol-d4) δ 8.46 (s, 1H), 8.09 (s, 1H), 6.98 (d, J = 8.3 Hz, 1H), 6.81 (d, J = 8.3 Hz, 1H), 2.48 (s, 3H), 1.64 (s, 3H), 1.59 (s, 3H); Mass spectrum: C 21 H 17D7N5O [M+H] + Calculated: 369.2, Found: 369.3.

[0715] Example 294: Preparation of 3-(4-amino-7-methyl-6-(propan-2-yl-d7)-1,2-dihydro-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl-2-d)-2,4-dimethylphenol

[0716] To a reaction vial was added the compound from Example 289 (20.00 mg, 54.13 μmol), followed by PtO2(12.29 mg, 54.13 μmol), followed by MeOH (3 mL), followed by a catalytic amount of HCl (4.0 M in Ethanol) (1.97 mg, 54.13 μmol), followed by replacement of H2, followed by heating to 50 °C and stirring overnight, followed by monitoring the reaction, followed by reaction completion, followed by filtration, followed by concentration, to produce 16.8 mg of the compound from Example 294 in 82% yield. 1 H NMR (600 MHz, Methanol-d4) δ 7.97 (s, 1H), 6.99 (d, J = 8.3 Hz, 1H), 6.82 (d, J = 8.3 Hz, 1H), 4.57 (d, J = 14.8 Hz, 1H), 2.36 (s, 3H), 1.77 (s, 3H), 1.72 (s, 3H); Mass Spectrum: C 21 H 18 D8N5O [M+H] + Calculated: 372.3, Found: 372.3.

[0717] Example 295: Preparation of 3-(4-amino-7-methyl-6-(propan-2-yl-d7)-1,2-dihydro-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl-2,2-d2)-2,4-dimethylphenol

[0718] To a reaction vial was added the compound from Example 289 (50.00 mg, 135.32 μmol), followed by PtO2(30.73 mg, 135.32 μmol), followed by MeOH (3 mL), followed by a catalytic amount of HCl (4.0 M in Ethanol) (4.9 mg, 135.32 μmol), followed by replacement of D2, followed by heating to 50 °C and stirring overnight, followed by monitoring the reaction, followed by reaction completion, followed by filtration, followed by concentration, followed by purification by preparative HPLC to produce 21.1 mg of the compound from Example 295 in 74% yield. 1H NMR (600 MHz, Methanol-d4) δ 7.98 (s, 1H), 7.01 (d, J = 8.3 Hz, 1H), 6.85 (d, J = 8.3 Hz, 1H), 2.38 (s, 3H), 1.79 (s, 3H), 1.74 (s, 3H); Mass: C 21 H 17 D9N5O [M+H] + Calculated: 373.3, Found: 373.3.

[0719] Examples 296-304 of Table 20 were prepared according to the synthetic method of Examples 294-295.

[0720] Table 20: Compounds of Examples 296-304

[0721] Example 305: Preparation of 3-(4-amino-1,7-dimethyl-6-(1,1,1 - trifluoropropan-2-yl)-1,2-dihydro-9H-pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)- 2,4-dimethylphenol

[0722] Compound R-16 was synthesized according to the synthetic method of Reference Example 145.

[0723] Step 1 : Synthesis of compound R-17

[0724] Into a reaction vial, was placed R-16 (30 mg, 69.86 μmol), followed by Pt02(23.80 mg, 104.79 μmol), then MeOH (5 mL), catalytic amount of HC1 (4.0 M in Ethanol) (5.09 mg, 139.72 μmol) to replace H2, heated to 50 °C and stirred overnight. The reaction was monitored and upon completion, filtered and concentrated to give crude compound R-17 which was used directly in the next step.

[0725] Step 2: Synthesis of compound R-18

[0726] Into a reaction vial, was placed R-17 (25 mg, 57.94 μmol), followed by MeOH (3 mL) to dissolve, then iodomethane (8.22 mg, 57.94 μmol), K2C03(16.02 mg, 115.89 μmol) was added dropwise. The reaction was stirred at room temperature and monitored. Upon completion, it was concentrated, filtered with a small amount of methanol to give the filtrate which was concentrated to give 1 mg of the target compound R-18 in 3.8% yield.

[0727] Step 3: Preparation of the compound of Example 305

[0728] To a reaction vial was added R-18 (0.8 mg, 1.80 pmol), then DCM (2 mL) to dissolve, BBr3 (10.77 pmol) was added slowly, the reaction was stirred at room temperature for 4 h, after the reaction was completed, methanol was added to quench, saturated NaHC03 solution was added to adjust to basic, DCM was extracted, the aqueous phase was extracted with EA again, the organic layers were combined and concentrated, and 0.4 mg of the compound of Example 305 was obtained by prep-HPLC purification with a yield of 50%. 1 H NMR (400 MHz, Methanol-d4) d 8.36 (s, 1H), 6.94 (dd, J = 8.3, 2.6 Hz, 1H), 6.78 (d, J = 8.3 Hz, 1H), 4.83 - 4.88 (m, 2H), 3.89 - 3.99 (m, 1H), 3.24 (s, 3H), 2.46 (s, 3H), 1.71 (dd, J = 23.1, 12.6 Hz, 6H), 1.56 (d, J = 7.1 Hz, 3H); Mass: C 22 H 25 F3N5O [M+H] + Calculated: 432.2, Found: 432.2.

[0729] Example 306: Preparation of 2-(4-amino-9-(3-hydroxy-2,6-xylyl)-7-methyl-9H- pyrido[3',2':4,5]pyrrolo[2,3-d]pyrimidin-6-yl)propane-1,3-diol

[0730] Step 1: Synthesis of compound S-2

[0731] To a reaction vial was added F-7 (300 mg, 727.66 pmol), S-1 (267.84 mg, 1.46 mmol), DPPF PdCl2 (532.43 mg, 727.66 pmol), K2C03 (301.71 mg, 2.18 mmol), then H20 (1 mL), 1,4-dioxane (2 mL) to dissolve, N2 was replaced, and the reaction was stirred at 110 °C for 1 h after microwave heating, the reaction was completed, EA was extracted, the organic layer was concentrated, and 200 mg of compound S-2 was obtained by column chromatography with a yield of 70%.

[0732] Step 2: Synthesis of compound S-3

[0733] A reaction vial was charged with S-2 (200 mg, 513.55 µmol), anhydrous THF (2 mL), replaced with N2, and BH3-THF solution (5.14 mmol) was added dropwise under ice water bath. After the addition was completed, the reaction was stirred at room temperature for 2.5 h. After the reaction was completed, ice water was added and stirred for 10 min. Then 3N NaOH solution (5.14 mmol) and H2O2 solution (5.14 mmol) were added and stirred for 30 min. After the reaction was completed, extraction was performed with EA to obtain an organic layer, which was concentrated and purified by column chromatography to obtain 90 mg of the target compound S-3 with a yield of 43%.

[0734] Step 3: Preparation of the compound of Example 306

[0735] A reaction vial was charged with S-3 (10 mg, 24.54 µmol), and then DCM (2 mL) was added for dissolution. BBr3 (147.25 µmol) was slowly added, and the reaction was stirred at room temperature for 4 h. After the reaction was completed, methanol was added for quenching. Saturated NaHCO3 solution was added to adjust the solution to be alkaline. Extraction was performed with DCM, and the aqueous phase was extracted with EA. The combined organic layers were concentrated and purified by preparative HPLC to obtain 6.1 mg of the compound of Example 306 with a yield of 62%. 1 H NMR (600 MHz, Methanol-d4) δ 8.77 (s, 1H), 8.47 (s, 1H), 7.12 (d, J = 8.3 Hz, 1H), 6.98 (d, J = 8.3 Hz, 1H), 4.02–4.05 (m, 2H), 3.94–3.97 (m, 2H), 3.54 (p, J = 6.6 Hz, 1H), 2.72 (s, 3H), 1.78 (s, 3H), 1.71 (s, 3H); Mass spectrum: C 21 H 24 N5O3 [M+H] + Calcd: 394.2, Found: 394.2.

[0736] Test Example 1: Enzyme activity test of the compound

[0737] PKMYT1 is a typical serine / threonine protein kinase, and the activity of the kinase is reflected by the amount of ADP generated per unit of time. First, the recombinant human PKMYT1 full-length protein with an N-terminal GST tag was purchased from ThermoFisherTM company (item number A32902), which was expressed by an insect SF9 system and purified by standard molecular sieve. In order to obtain the half-effective inhibition concentration (IC50) of the compound, the activity of the enzyme was first determined by using ATP as a substrate and ADP as a product. The activity of the enzyme was determined by using the following formula: Activity = (OD450-OD650) / (min x mg protein) 50), the experiment first uses a 96-well semi-skirt PCR plate (ThermoFisherTM, AB1400L) to incubate the above-mentioned GST-PKMYT1 recombinant protein at a final reaction concentration of 18.5nM with a solvent blank as a positive reference and a buffer without the recombinant protein as a negative reference, using an 8-point concentration gradient of the inhibitor at a final concentration of 500nM, 166.67nM, 55.56nM, 18.52nM, 6.17nM, 2.06nM, 0.69nM, and 0.23nM in a kinase buffer (70mM HEPES, 3mM MgCl2, 3mM MnCl2, 50μg / ml PEG2000, 1.2mM DTT, and 3mM sodium orthovanadate, pH7.4) and pre-incubated for 15 minutes in a constant temperature incubation shaker at 25℃ and 300RPM. Then, the kinase reaction is started by adding ATP at a final concentration of 10μM, and the kinase activity reaction conditions are accurately performed for 1 hour of enzymatic reaction in a constant temperature incubation shaker at 30℃ and 300RPM. Then, the final reaction sample is transferred to a standard 384-well plate (Greiner, 781075), and the chemical light quantum number corresponding to the ADP generated is obtained by the ADP-GloTM kinase detection kit (PromegaTM, V9101) through a two-step standard experimental procedure. The above light quantum number signal is detected by a BioTek Synergy Neo2 enzyme label instrument (Agilent) (detection parameters: signal reading mode is top-reading chemiluminescence mode, temperature setting is 25℃, horizontal medium intensity shaking plate for 15s, integration time is 40ms, delay time is 20ms, signal gain is 255, signal reading height is automatic. After obtaining the above signal value, all readings are deducted from the average chemiluminescence signal value without protein (i.e., the average reading value is used as the signal background). Then, the signal value of each compound gradient is converted into the relative enzyme activity parameter by taking the average reading value of the group containing only the recombinant protein and not containing the compound as 100% enzyme activity, and finally the above value is substituted into the GraphPad Prism software for statistical analysis. In order to obtain the inhibition IC50 of the compound, the enzyme-inhibition (variable slope four-parameter formula fitting) is used for analysis, and the IC50 value and 95% confidence interval are calculated to obtain the average IC50 value. As part of the standardized system quality control, the same compound in the same batch is used as a reference to ensure that the IC50 value deviation from the average value is not more than 50%. At the same time, the 100% enzyme signal reading value interval should be greater than 3 times and less than 10 times the 0% signal original value. The test results are shown in Table 21, and it can be seen that the compounds of the present application have strong PKMYT1 inhibitory effect and can be used as drugs for treating PKMYT1 overexpression related diseases.

[0738] Table 21: Inhibition activity (IC50) of part of the compounds of the examples on PKMYT1 50 ) Note: " / " means not detected.

[0739] Test Example 2: Compound cell activity test:

[0740] 1.1 Experimental materials

[0741] 1.1.1 Cell lines

[0742] Human breast cancer cells HCC1569 and human ovarian cancer cells OVCAR-3 were purchased from China Center for Type Culture Collection (CCTCC).

[0743] 1.1.2 Main experimental instruments

[0744] 1.1.3 Main reagents

[0745] 2.1 Experimental methods

[0746] 2.1.1 Cell culture

[0747] HCC1569 cells were cultured in RPMI 1640 medium containing 10% fetal bovine serum and 1% penicillin-streptomycin, and OVCAR3 cells were cultured in RPMI 1640 medium containing 20% fetal bovine serum and 1% penicillin-streptomycin. All cell strains were cultured in a sterile incubator at 37°C and 5% CO2, and cell passage, medium replacement and other operations were carried out in a sterile table in strict accordance with the sterile principle.

[0748] 2.1.2 Cell recovery

[0749] The cells were taken out from the liquid ammonia tank and melted in a 37°C water bath. Centrifuge at 1000 rpm / min for 5 min, resuspend the cells, and transfer to a cell culture dish containing complete culture medium. Continue to culture in a 5% CO2, 37°C cell incubator.

[0750] 2.1.3 Cell freezing

[0751] The cell culture used double-antibody-free medium, or at least the medium was changed to double-antibody-free medium 1 day before freezing; the cells were frozen when the cultured cells were in the logarithmic growth phase, the adherent cells were routinely digested into single cells, centrifuged with complete culture medium, the suspended cells were directly centrifuged at 1000 rpm / min for 5 min, the supernatant was removed, the precipitate was resuspended with freezing solution (90% fetal bovine serum and 10% DMSO), and then transferred to -80°C (freezing box) for 24 hours before being stored in liquid nitrogen.

[0752] 2.1.4 Tumor cell EC50assay

[0753] (1) Day -1 : 300 cells were seeded per well in a final volume of 50 μl (cell concentration of 6000 cells / ml) and incubated in the cell incubator for 24 hours to allow cell attachment.

[0754] (2) Day 0: Serial dilutions of test compound were added using a multichannel pipettor. The compound was serially diluted in culture medium containing 0.1 v / v % DMSO to give final concentrations of (in nM): 10000.00, 3246.75, 1054.14, 342.25, 111.12, 36.08, 11.71, 3.80, respectively. The diluted compound was added to the 96 well plates and incubated with the cells for 4 days.

[0755] (3) Day 4: The original culture medium was replaced with fresh culture medium containing serial dilutions of test compound. 185 μl medium + 0.37 μl 10 mM test compound stock (highest concentration: 20 μM). 60 μl medium + 125 μl medium containing 0.1 % DMSO test compound serial dilution (8 concentrations + blank control)

[0756] (4) Day 7: The 96 well plates were removed from the cell incubator and equilibrated to room temperature (RT) for 30 minutes. The medium was aspirated from all wells using an 8 channel vacuum manifold and diluted CTG reagent was added (diluted 1 :4 with PBS). The plates were shaken on a plate shaker at 1000 rpm for 5 minutes at 25°C and incubated at room temperature for 10 minutes in the dark to stabilize the CTG signal.

[0757] (5) The multi-function microplate reader was set to the following parameters for fluorescence endpoint detection: Integration Time 20 msec, Filter Set 1, Emission Full Spectrum, Optical Detection Top, Read Speed Normal, Read Height 1 mm, Gain 135, Temperature 25°C. The results are shown in Table 22.

[0758] Table 22: Effect Concentration (EC50) of Example Compounds on Tumor Cells 50 ) Note: " / " means not tested.

[0759] Test Example 3: Rat skin rash model test

[0760] 1. Animal grouping

[0761] The BN rats were randomly grouped by scientific method.

[0762] 2. Dosing

[0763] (1) 100% DMSO to dissolve the compound into a certain concentration of mother liquor, stored at -20°C. On the day of administration, add cosolvent and mix well, ultrasonic dissolution in 50°C water bath. The cosolvent is a mixture of 0.5% CMC-Na and 15% Kolliphor HS15.

[0764] (2) According to the body weight of rats, PO / BID administration at a volume of 10 μl / g.

[0765] 3. Index detection and tissue sample collection

[0766] (1) General state observation of animals: At random time points every day, the general living conditions of BN rats were observed. The observation contents mainly include: spirit, movement, hair, skin, and abnormal conditions were photographed and recorded.

[0767] (2) The body weight of each BN rat was recorded three times a week, and the food intake of each group of animals was recorded.

[0768] (3) On the 7th day of the experiment, 50 μL of whole blood was taken from the mandibular vein of the BN rats for blood routine determination. After the end of the experiment, one rat from each group was euthanized, and the skin of the hindpaws and back was preserved in fixative for HE staining by Saviol. The whole blood of the animals was tested for blood routine by animal automatic blood cell analyzer; the plasma compound content of the animals was analyzed by LC-MS.

[0769] 4. Statistical analysis

[0770] The experimental data are expressed as mean ± standard error (mean ± SEM), and the experimental data are analyzed and processed by Graphpad Prism 8.0 software. When P<0.05, the results are significantly different. Representative patent compounds were not observed to have abnormal skin rash phenotype in the rat skin rash model, the biochemical indicators were normal, and showed good safety.

[0771] Test Example 4: In vivo efficacy test in mice:

[0772] 1. HCC1569 cell collection

[0773] (1) Remove the old culture medium, wash the cells once with PBS solution. Discard the wash, add 1 ml of cell digestion solution and incubate in the incubator for 15-20 min.

[0774] (2) After most of the cells are detached from the surface of the culture dish, add 5 ml PBS / dish, gently blow, form a single cell suspension, centrifuge (750 rpm, 3 min, 4°C).

[0775] (3) Discard the supernatant, resuspend the cell pellet in PBS to a final volume of 45 ml.

[0776] (4) Take 10 μl of the resuspended cell suspension into another 1.5 ml Eppendorf tube, add equal volume (10 μl) of 0.4% trypan blue solution, mix well by pipetting. Take 10 μl to a cell counting slide and count.

[0777] (5) Centrifuge (750 rpm, 3 min, 4°C), add pre-cooled PBS solution (4°C fridge pre-cooled) according to the counting result and the number of cells required

[0778] (6) Add 2x growth factor supplement

[0779] (7) Matrigel is thawed in advance at 4°C fridge overnight, add equal volume of matrigel and mix well (i.e. Vcells: Vmatrigel = 1:1), aliquot the mixed cell-matrigel suspension into 1.5 ml Eppendorf tubes.

[0780] 2. Subcutaneous inoculation of HCC1569 cells

[0781] (1) Shave the right flank and half of the back of the mice 1-2 days in advance, and mark the right ear with ear tags. Inject subcutaneously (SC) each NCG mouse with a final volume of 5 x 10 6 cells / 100 μl / mouse of HCC1569 cell-matrigel suspension.

[0782] (2) After inoculation of the cells, observe the tumor growth every two days, and measure the tumor size of the mice (volume = 0.52 x length x width2) with a vernier caliper. When the matrigel is mostly absorbed and the tumor volume grows to 100-150 mm 3 , randomly group the mice scientifically, and administer PO / BID.

[0783] 3. Drug administration

[0784] (1) Dissolve the compounds in 100% DMSO to a certain concentration of stock solution, aliquot and store at -20°C. On the day of administration, add cosolvent and mix well, and then dissolve in a 50°C water bath. The cosolvent is a mixture of 0.5% CMC-Na and 15% Kolliphor HS15.

[0785] (2) According to the body weight of the mice, administer PO / BID at a volume of 10 μl / g.

[0786] 4. Data collection and analysis

[0787] Mouse body weight was recorded weekly post-dose, Monday, Wednesday, Friday, and mouse tumor size was measured using vernier calipers (volume = 0.52 x length x width2). Representative patent compounds showed excellent tumor inhibition activity in the mouse HCC1569 cell subcutaneous inoculation model.

[0788] The above only the preferred embodiments of the present application, and is not intended to limit the present application, for those skilled in the art, the present application can have various modifications and changes. Any modification, equivalent replacement, improvement, etc. within the spirit and principles of the present application, should be included in the scope of protection of the present application.

Claims

1. A compound of formula (I), a pharmaceutically acceptable salt, stereoisomer, isotopic derivative, solvate or prodrug thereof, wherein Ring A is selected from a six-membered unsaturated nitrogen-containing heterocyclic ring, said six-membered unsaturated nitrogen-containing heterocyclic ring optionally containing one or two additional heteroatoms selected from N, O and S, and optionally substituted by one or more R A Substituted, where R A each independently selected from -D, -OH, halogen, oxo (=O), -CN, C1-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, 3-10 membered cycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl, -NHR1, -NHC(O)R1, -NHC(O)NHR1, or -NHSO2R1, and wherein R1 is selected from -H, -D, -OH, -NH2, C1-10 alkyl, or 3-10 membered cycloalkyl; Ring B is selected from a six-membered unsaturated nitrogen-containing heterocyclic ring, which optionally contains one or two additional heteroatoms selected from N, O and S, and is optionally substituted by one or more R B Substituted, where R B Each independently selected from -(C≡C) m -R2, -O-(CH2) n -R2, -SO2R2, -C(O)NR3R4, or -SO2NR3R4, wherein m is selected from 0 or 1, n is an integer selected from 0 to 3, and wherein R2 is selected from -D, halogen, -CN, -OH, -NR3R4, optionally replaced by one or more R s1 Substituted C1-10 alkyl, optionally substituted by one or more R s1 Substituted C2-10 alkenyl, optionally substituted with one or more R s1 Substituted C2-10 alkynyl, optionally substituted by one or more R s1 Substituted C1-10 alkoxy, optionally substituted by one or more R s2 substituted 3-10 membered cycloalkyl, optionally substituted by one or more R s2 substituted 3-10 membered cycloalkenyl, optionally substituted with one or more R s2 substituted 3-10 membered heterocycloalkyl, optionally substituted by one or more R s2 substituted 3-10 membered heterocycloalkenyl, optionally substituted by one or more R s2 substituted 5-10 membered aryl, or optionally substituted with one or more R s2 substituted 5-10 membered heteroaryl, wherein R3 and R4 are each independently selected from -H, -D, -OH, -CN, optionally substituted by one or more R s1 Substituted C1-5 alkyl, optionally substituted by one or more R s1 Substituted C2-10 alkenyl, optionally substituted with one or more R s1 substituted 3-10 membered cycloalkyl, optionally substituted by one or more R s1 substituted 5-10 membered heteroaryl, -C(O)-R5 or -(CH2) p -O-R5, wherein p is an integer selected from 0 to 5, wherein R5 is selected from -H, -D or C1-5 alkyl, R s1 are each independently selected from -D, -OH, halogen, -CN, -NH2, C1-5 alkyl, 3-10 membered cycloalkyl, C1-5 alkoxy or -C(O)-R5, and R s2 Each independently selected from -D, -OH, halogen, -CN, -NR3R4, oxo (=O), -C(O)-R5, -C(O)-NR3R4, optionally substituted by one or more R s1 Substituted C1-5 alkyl, or optionally substituted by one or more R s1 substituted 5-10 membered heterocycloalkyl; Ring C is selected from optionally substituted by one or more R s1 substituted phenyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolyl or indazolyl; Q and U are each independently selected from N, C or CH, and Indicates a single bond or a double bond.

2. The compound according to claim 1, or its pharmaceutically acceptable salt, stereoisomer, isotope derivative, solvate or prodrug, wherein: Ring A is selected from the group consisting of A Replaces:

3. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt, stereoisomer, isotope derivative, solvate or prodrug thereof, wherein: Ring A is selected from: wherein R6, R7, and R8 are each independently selected from -H or R A , preferably, wherein the * carbon is connected to U, further preferably, wherein R6, R7, and R8 are not all -H; and / or R6 is selected from -H, -D, -OH, halogen, -CN, C1-5 alkyl, C2-6 alkenyl, C2-6 alkynyl, 3-8 membered cycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl, -NHR1, -NHC(O)R1, -NHC(O)NHR1, or -NHSO2R1, and wherein R1 is selected from -H, -D, -OH, -NH2, C1-5 alkyl, or 3-8 membered cycloalkyl; and / or R7 is selected from -H, -D, -OH, halogen, -CN, C1-5 alkyl, C2-6 alkenyl, C2-6 alkynyl, 3-8 membered cycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl, -NHR1, -NHC(O)R1, -NHC(O)NHR1, or -NHSO2R1, and wherein R1 is selected from -H, -D, -OH, -NH2, C1-5 alkyl, or 3-8 membered cycloalkyl; and / or R8 is selected from -H, -D, -OH, halogen, -CN, C1-5 alkyl, C2-6 alkenyl, C2-6 alkynyl, 5-10 membered aryl, 5-10 membered heteroaryl, -NHR1, -NHC(O)R1, -NHC(O)NHR1, or -NHSO2R1, and wherein R1 is selected from -H, -D, -OH, -NH2 and C1-5 alkyl.

4. A compound according to any one of the preceding claims, or a pharmaceutically acceptable salt, stereoisomer, isotopic derivative, solvate or prodrug thereof, wherein Ring B is selected from the group consisting of B Replaces: Preferably, ring B is selected from Where * carbon is connected to U, and R9, R 10 、R 11 Each independently selected from -H or R B , Preferably, wherein R9, R 10 、R 11 Any two of them are not both -H; and / or Among them, R9, R 10 、R 11 Each independently selected from -H, -(C≡C) m -R2, -O-(CH2) n -R2, -SO2R2, -C(O)NR3R4, or -SO2NR3R4, wherein m is selected from 0 or 1, n is an integer selected from 0 to 3, and wherein R2 is selected from -D, halogen, -CN, -OH, -NR3R4, optionally replaced by one or more R s1 Substituted C1-5 alkyl, optionally substituted by one or more R s1 Substituted C2-6 alkenyl, optionally substituted by one or more R s1 Substituted C2-6 alkynyl, optionally substituted by one or more R s1 Substituted C1-5 alkoxy, optionally substituted by one or more R s2 substituted 3-8 membered cycloalkyl, optionally substituted by one or more R s2 substituted 3-8 membered cycloalkenyl, optionally substituted with one or more R s2 substituted 3-8 membered heterocycloalkyl, optionally substituted by one or more R s2 substituted 3-8 membered heterocycloalkenyl, optionally substituted by one or more R s2 substituted 5-10 membered aryl, or optionally substituted with one or more R s2 substituted 5-10 membered heteroaryl, wherein R3 and R4 are each independently selected from -H, -D, -OH, -CN, optionally substituted by one or more R s1 Substituted C1-5 alkyl, optionally substituted by one or more R s1 Substituted C2-6 alkenyl, optionally substituted by one or more R s1 substituted 3-8 membered cycloalkyl, optionally substituted by one or more R s1 substituted 5-10 membered heteroaryl, -C(O)-R5 or -(CH2) p -O-R5, wherein p is an integer selected from 0 to 5, wherein R5 is selected from -H, -D or C1-5 alkyl, R s1 are each independently selected from -D, -OH, halogen, -CN, -NH2, C1-5 alkyl, 3-8 membered cycloalkyl or -C(O)-R5, and R s2 Each independently selected from -D, -OH, halogen, -CN, -NR3R4, oxo (=O), -C(O)-R5, -C(O)-NR3R4, optionally substituted by one or more R s1 Substituted C1-5 alkyl, or optionally substituted by one or more R s1 Substituted 5-10 membered heterocycloalkyl.

5. The compound according to claim 4, or its pharmaceutically acceptable salt, stereoisomer, isotope derivative, solvate or prodrug, wherein R9 is selected from -H, -D, halogen, -CN, -OH, C1-5 alkyl optionally substituted by one or more halogen or -D, C2-6 alkenyl optionally substituted by one or more halogen or -D, or 3-8 membered cycloalkyl optionally substituted by one or more halogen or -D; and / or R 10 and R 11 Each independently selected from -H, -(C≡C) m -R2, -O-(CH2) n -R2, or -SO2R2, wherein m is selected from 0 or 1, n is selected from 0, 1 or 2, and wherein R2 is selected from -H, -D, halogen, -CN, -OH, C1-5 alkyl optionally substituted with one or more halogen, -D, -OH, or -CN, or C2-6 alkenyl optionally substituted with one or more halogen, -D, -OH, or -CN, or C2-6 alkynyl optionally substituted with one or more halogen, -D, -OH, or -CN, or C1-5 alkoxy optionally substituted with one or more halogen, -D, -OH, or -CN; or R 10 and R 11 Each independently selected from: -NR3R4, -O-(CH2) n -NR3R4, -C(O)NR3R4, or -SO2NR3R4, wherein n is an integer selected from 0 to 3, and wherein R3 and R4 are each independently selected from -H, -D, -OH, -CN, optionally substituted by one or more R s1 Substituted C1-5 alkyl, optionally substituted by one or more R s1 Substituted C2-6 alkenyl, optionally substituted by one or more R s1 substituted 3-8 membered cycloalkyl, optionally substituted by one or more R s1 substituted 5-10 membered heteroaryl, -C(O)-R5 or -(CH2) p -O-R5, wherein p is an integer selected from 0 to 5, R5 is selected from -H, -D or C1-5 alkyl, and wherein R s1 are each independently selected from -D, -OH, halogen, or -CN; Also preferably, wherein the substituent -NR3R4 is selected from the group consisting of -NHCH3, -NHCH2CH3, -NHCH2CH2CH3, -NHCH(CH3)2, -N(CH3)2, -N(CH2CH3)2, -N(CH2CH2CH3)2, -NHCH2CH2OCH3, -NHCH=CH2, -NHCH2CH=CH2, -NHCH=CHCH3, -N(CH=CH2)2, -N(CH2CH=CH2)2, -N(CH=CHCH3)2, 6. The compound according to claim 4, or its pharmaceutically acceptable salt, stereoisomer, isotope derivative, solvate or prodrug, wherein R 10 and R 11 Each independently selected from -H, -(C≡C) m -R2, -O-(CH2) n -R2, or -SO2R2, wherein m is selected from 0 or 1, n is selected from 0, 1 or 2, and wherein R2 is selected from: wherein q is an integer independently selected from 0 to 6.

7. The compound according to claim 4, or its pharmaceutically acceptable salt, stereoisomer, isotope derivative, solvate or prodrug, wherein: R 10 and R 11 Each independently selected from -H, -(C≡C) m -R2, -O-(CH2) n -R2, or -SO2R2, wherein m is selected from 0 or 1, n is selected from 0, 1 or 2, and wherein R2 is selected from optionally substituted with one or more -D, halogen (especially -F), -OH or oxo:

8. A compound according to any one of the preceding claims, a pharmaceutically acceptable salt, stereoisomer, isotopic derivative, solvate or prodrug thereof, wherein Ring C is selected from where R 12 、R 13 、R 14 、R 15 Each is independently selected from -H, -D, -OH, halogen, -CN, -NH2, C1-5 alkyl or -C(O)-R5, wherein R5 is selected from -H, -D or C1-5 alkyl; preferably, ring C is selected from 9. A compound according to any one of the preceding claims, a pharmaceutically acceptable salt, stereoisomer, isotopic derivative, solvate or prodrug thereof, wherein The compound has a structure represented by formula (IIA), formula (IIB), formula (IIC) or formula (IID): in R6 and R7 are each independently selected from -H, -D, -OH, halogen, -CN, C1-5 alkyl, C2-6 alkenyl, C2-6 alkynyl, 3-8 membered cycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl, -NHR1, -NHC(O)R1, -NHC(O)NHR1, or -NHSO2R1, and wherein R1 is selected from -H, -D, -OH, -NH2, C1-5 alkyl, or 3-8 membered cycloalkyl; R9, R 10 、R 11 Each independently selected from -H, -(C≡C) m -R2, -O-(CH2) n -R2, -SO2R2, -C(O)NR3R4, or -SO2NR3R4, wherein m is selected from 0 or 1, n is an integer selected from 0 to 3, and wherein R2 is selected from -D, halogen, -CN, -OH, -NR3R4, optionally replaced by one or more R s1 Substituted C1-5 alkyl, optionally substituted by one or more R s1 Substituted C2-6 alkenyl, optionally substituted by one or more R s1 Substituted C2-6 alkynyl, optionally substituted by one or more R s1 Substituted C1-5 alkoxy, optionally substituted by one or more R s2 substituted 3-8 membered cycloalkyl, optionally substituted by one or more R s2 substituted 3-8 membered cycloalkenyl, optionally substituted with one or more R s2 substituted 3-8 membered heterocycloalkyl, optionally substituted by one or more R s2 substituted 3-8 membered heterocycloalkenyl, optionally substituted by one or more R s2 substituted 5-10 membered aryl, or optionally substituted with one or more R s2 substituted 5-10 membered heteroaryl, wherein R3 and R4 are each independently selected from -H, -D, -OH, -CN, optionally substituted by one or more R s1 Substituted C1-5 alkyl, optionally substituted by one or more R s1 Substituted C2-6 alkenyl, optionally substituted by one or more R s1 substituted 3-8 membered cycloalkyl, optionally substituted by one or more R s1 substituted 5-10 membered heteroaryl, -C(O)-R5 or -(CH2) p -O-R5, wherein p is an integer selected from 0 to 5, wherein R5 is selected from -H, -D or C1-5 alkyl, R s1 are each independently selected from -D, -OH, halogen, -CN, -NH2, C1-5 alkyl, or -C(O)-R5, and R s2 Each independently selected from -D, -OH, halogen, -CN, -NR3R4, oxo (=O), -C(O)-R5, -C(O)-NR3R4, optionally substituted by one or more R s1 Substituted C1-5 alkyl, or optionally substituted by one or more R s1 substituted 5-10 membered heterocycloalkyl; R 12 、R 13 、R 14 、R 15 Each is independently selected from -H, -D, -OH, halogen, -CN, -NH2, C1-5 alkyl, 3-8 membered cycloalkyl, or -C(O)-R5, wherein R5 is selected from -H, -D or C1-5 alkyl.

10. A compound according to any one of the preceding claims, a pharmaceutically acceptable salt, stereoisomer, isotopic derivative, solvate or prodrug thereof, wherein The compound has a structure represented by formula (IIA-1) or (IIA-2): in, R7 is hydrogen or deuterium; R 15 is hydrogen or halogen (preferably, F); R 11 is hydrogen, CN or methyl or ethyl optionally substituted by one or more deuterium groups; R 10 Selected from: - halogen (such as F, Cl or Br, preferably Br), - optionally replaced by one or more R k Substituted C 2-3 Alkyl (such as ethyl, n-propyl or isopropyl), C 2-3 alkenyl (e.g., vinyl, propenyl, allyl, isopropenyl), cyclopropyl, 5-6 membered heterocycloalkyl containing 1-2 heteroatoms selected from O or N, 5-6 membered heterocycloalkenyl containing 1-2 heteroatoms selected from O or N, 5-6 membered heteroaryl containing 1-2 heteroatoms independently selected from O, N or S, wherein R k are each independently selected from halogen (preferably, F or Cl), hydroxyl, cyano or deuterium, - unsubstituted phenyl, or --NR3R4, wherein R3 and R4 are each independently selected from H, C optionally substituted with 1 to 7 deuteriums 1-3 Alkyl or cyclopropyl; The conditions are: i) the heterocycloalkenyl group contains only one double bond; ii) When R 11 When it is CN, R 10 Selected from unsubstituted C 2-3 Alkyl or unsubstituted C 2-3 alkenyl; iii) When R 10 When it is a 5-6 membered heteroaryl group containing two heteroatoms independently selected from O, N or S, the two heteroatoms are not N at the same time; iv) When R 10 When -NR3R4, R3 and R4 are not C optionally substituted by deuterium at the same time 2-3 alkyl, or cyclopropyl optionally substituted by deuterium, or C 2-3 Alkyl and cyclopropyl; v) When R 10 When it is a substituted or unsubstituted 5-6 membered heterocycloalkyl group containing one N atom, it is connected to the pyridine ring of formula (IIA-1) or formula (IIA-2) through the N atom; or vi) When R 10 When it is a substituted or unsubstituted 5-6 membered heteroaryl group containing one nitrogen atom, it is not connected to the pyridine ring of formula (IIA-1) or formula (IIA-2) via the nitrogen atom.

11. The compound according to claim 10, or a pharmaceutically acceptable salt, stereoisomer, isotope derivative, solvate or prodrug thereof, wherein when R 11 When R is hydrogen or methyl optionally substituted by one or more deuterium groups, 10 Selected from: -F, Cl, Br, - optionally substituted by one or more R each independently selected from halogen (preferably F or Cl), hydroxyl, cyano or deuterium k Substituted ethyl, isopropyl, vinyl, propenyl, allyl, isopropenyl, cyclopropyl, - optionally substituted by one or more R each independently selected from halogen (preferably F or Cl) or deuterium k Substituted tetrahydrofuranyl, tetrahydropyrrolyl, tetrahydropyranyl, piperidinyl, - optionally substituted by one or more R each independently selected from halogen (preferably F or Cl) or deuterium k Replaced - optionally substituted by one or more R each independently selected from halogen (preferably F or Cl) or deuterium k Substituted pyrrolyl, furyl, thienyl, imidazolyl, oxazolyl, isoxazolyl, pyrazolyl, thiazolyl, pyridinyl, e.g. - unsubstituted phenyl, or --NR3R4, wherein R3 and R4 are each independently selected from H, methyl, ethyl, n-propyl, isopropyl or cyclopropyl optionally substituted with 1 to 7 deuterium atoms; Preferably, when R 11 When R is hydrogen or methyl optionally substituted by 1 to 3 deuterium atoms, 10 Selected from: -Br， - 1 hydroxyl or cyano group and / or 1 to 7 R groups independently selected from F or deuterium k Substituted ethyl, isopropyl, vinyl, propenyl, allyl, isopropenyl, cyclopropyl, - unsubstituted ethyl, isopropyl, vinyl, propenyl, allyl, isopropenyl, cyclopropyl, - unsubstituted tetrahydrofuranyl, tetrahydropyrrolyl, tetrahydropyranyl, piperidinyl, -Unsubstituted - unsubstituted pyrrolyl, furyl, thienyl, imidazolyl, oxazolyl, isoxazolyl, pyrazolyl, thiazolyl, pyridyl, e.g. - unsubstituted phenyl, or --NR3R4, wherein R3 and R4 are each independently selected from H, methyl, ethyl, isopropyl or cyclopropyl optionally substituted with 1 to 7 deuteriums.

12. The compound according to claim 10, or a pharmaceutically acceptable salt, stereoisomer, isotope derivative, solvate or prodrug thereof, wherein when R 11 When R is an ethyl group optionally substituted with 1 to 5 deuterium groups, 10 Selected from 1 to 4 R k substituted pyrrolyl, furyl, thienyl, imidazolyl, oxazolyl, isoxazolyl, pyrazolyl, thiazolyl, or optionally substituted with 1 to 7 R k Substituted isopropyl or isopropenyl, wherein R k are each independently selected from halogen (preferably, F or Cl) or deuterium.

13. The compound according to claim 1, or a pharmaceutically acceptable salt, stereoisomer, isotopic derivative, solvate or prodrug thereof, wherein the compound is selected from the group consisting of:

14. The compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt, stereoisomer, isotopic derivative, solvate or prodrug thereof, wherein The compound is enriched in atropisomers of Formula (IIAi), Formula (IIBi), Formula (IICi) or Formula (IIDi):

15. The compound according to any one of claims 1 to 12, or a pharmaceutically acceptable salt, stereoisomer, isotopic derivative, solvate or prodrug thereof, wherein The compound is enriched in atropisomers of Formula (IIAii), Formula (IIBii), Formula (IICii) or Formula (IIDii):

16. The compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer, isotopic derivative, solvate, or prodrug thereof, wherein The compound is selected from 17. A pharmaceutical composition comprising the compound according to any one of claims 1 to 16 or a pharmaceutically acceptable salt, stereoisomer, isotopic derivative, solvate or prodrug thereof, and a pharmaceutically acceptable carrier or excipient.

18. Use of the compound according to any one of claims 1 to 16 or a pharmaceutically acceptable salt, stereoisomer, isotope derivative, solvate or prodrug thereof in the preparation of a medicament for inhibiting the activity of PKMYT1 in cells.

19. The use of claim 18, wherein the drug is used to treat cancer; preferably, PKMYT1 is overexpressed in the cancer; further preferably, the cancer is selected from breast cancer, ovarian cancer, uterine cancer, gastric cancer, lung cancer (especially non-small cell lung cancer), hepatocellular carcinoma, glioblastoma, neuroblastoma, colorectal cancer, or esophageal cancer.

20. A method for inhibiting the activity of PKMYT1 in a cell overexpressing PKMYT1, the method comprising contacting the cell with a compound according to any one of claims 1 to 16 or a pharmaceutically acceptable salt, stereoisomer, isotopic derivative, solvate or prodrug thereof, or a pharmaceutical composition according to claim 17.

21. The method of claim 20, wherein the cell is a mammalian cell; and / or the method is an in vitro non-therapeutic method.

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