Compound targeting protein modulator and use thereof

Abstract

Disclosed are a compound targeting a protein modulator and a use thereof. The compound is a compound represented by formula (I), or a pharmaceutically acceptable salt thereof. The compound of the present invention has a good inhibition effect or protein degradation effect on a KRAS mutant.

Description

A compound of targeted protein regulator and its application

[0001] This application claims priority to Chinese Patent Application No. 2023115320552, filed on November 15, 2023, and Chinese Patent Application No. 202410503372X, filed on April 24, 2024. The entire text of the above-mentioned Chinese patent application is incorporated herein by reference. Technical Field

[0002] The present invention relates to a compound of a targeted protein regulator and application thereof. Background Art

[0003] RAS represents a group of closely related monomeric globular proteins (21 kDa molecular weight) with 189 amino acids that are associated with the plasma membrane and bind GDP or GTP. RAS acts as a molecular switch. When RAS contains bound GDP, it is in a resting or closed position and is "inactive." In response to cell exposure to certain growth-promoting stimuli, RAS is induced to exchange its bound GDP for GTP. When bound to GTP, RAS is "opened" and can interact with and activate other proteins (its "downstream targets"). The RAS protein itself has a very low intrinsic ability to hydrolyze GTP back to GDP, thereby turning itself into a closed state. Turning off RAS requires an exogenous protein called GTPase activating protein (GAP), which interacts with RAS and greatly accelerates the conversion of GTP to GDP. Any mutation in RAS that affects its ability to interact with GAP or convert GTP back to GDP will result in prolonged protein activation, and therefore prolonged signals transmitted to cells telling them to continue growing and dividing. Because these signals cause cell growth and division, overactivated RAS signaling can ultimately lead to cancer.

[0004] Structurally, the RAS protein contains a G domain responsible for the enzymatic activity of RAS - guanine nucleotide binding and hydrolysis (GTPase reaction). It also contains a C-terminal extension called the CAAX box, which can be post-translationally modified and is responsible for targeting the protein to the membrane. The G domain is approximately 21-25 kDa in size and contains a phosphate binding loop (P-loop). The P-loop represents the pocket in the protein that binds nucleotides, and this is the rigid part of the domain with conserved amino acid residues that are necessary for nucleotide binding and hydrolysis (glycine 12, threonine 26, and lysine 16). The G domain also contains the so-called switch I region (residues 30-40) and switch II region (residues 60-76), which are both dynamic parts of the protein and are often referred to as a "spring-loaded" mechanism due to the ability of the dynamic part to switch between a resting and loaded state. The main interaction is the hydrogen bond formed by threonine-35 and glycine-60 with the γ-phosphate of GTP, which enables the switch 1 region and the switch 2 region to maintain their active conformations, respectively. Following hydrolysis of GTP and release of phosphate, both relax into the inactive GDP conformation.

[0005] The most notable members of the RAS subfamily are HRAS, KRAS, and NRAS, which are primarily implicated in many types of cancer. However, there are numerous other members, including DIRAS1; DIRAS2; DIRAS3; ERAS; GEM; MRAS; NKIRAS1; NKIRAS2; NRAS; RALA; RALB; RAP1A; RAP1B; RAP2A; RAP2B; RAP2C; RASD1; RASD2; RASL10A; RASL10B; RASL11A; RASL11B; RASL12; REM1; REM2; RERG; RERGL; RRAD; RRAS, and RRAS2.

[0006] Mutations in any of the three major isoforms of the RAS gene (HRAS, NRAS, or KRAS) are among the most common events in human tumor formation. Approximately 30% of all human tumors were found to carry some mutation in the RAS gene. Notably, KRAS mutations were detected in 25%-30% of tumors. In contrast, the rates of oncogenic mutations in NRAS and HRAS family members were much lower (8% and 3%, respectively). The most common KRAS mutations were found at residues G12 and G13 in the P-loop and at residue Q61. Among tumor-associated KRAS G12 mutations, KRAS G12D had the highest mutation rate, at approximately 40%.

[0007] Given the importance of KRAS aberrant activation in cancer progression and the prevalence of KRAS mutations in human cancers, KRAS has long been a target of interest for drug developers. Despite progress in this area, there remains a need for improved inhibitors of the KRAS G12D mutant protein.

[0008] In recent years, people have taken advantage of the functional characteristics of the ubiquitin proteasome pathway to specifically degrade protein substrates and constructed proteolysis-targeting chimeras (PROTACs). PROTACs are compounds with two heterofunctional ligands connected by a linker: one ligand targets the protein of interest (POI), while the other ligand specifically recruits the E3 ligase. When PROTAC binds to the E3 ligase and the target protein, a ternary complex is formed. By hijacking the E3 ligase, PROTAC places the POI in a favorable spatial position to promote its ubiquitination, thereby selectively reducing the level of the target protein. The advantage of this approach is that PROTAC can catalyze multiple rounds of degradation of the target protein, which is the biggest difference between PROTAC molecules and small molecule inhibitors.

[0009] CN110684015A discloses a PROTAC molecule targeting ALK. This molecule was successfully prepared and effectively targets the target protein and reduces ALK levels in cells. It also exhibits good anti-tumor activity in vitro and in vivo, and has low toxicity to normal cells, meeting the requirements of high efficiency and low toxicity. PROTAC molecules targeting KRAS are also a hot topic for research and development by those skilled in the art. Summary of the Invention

[0010] The present invention aims to overcome the limitation of the existing technology in that there are relatively few compounds that can target and degrade or inhibit KRAS mutants. To this end, a compound that acts as a targeted protein regulator and its use are provided. The compound of the present invention has a good inhibitory effect or protein degradation effect on KRAS mutants.

[0011] The present invention solves the above technical problems through the following methods.

[0012] The present invention provides a compound as shown in Formula I, or a pharmaceutically acceptable salt thereof:

[0013] Among them, in the compound shown in formula I:

[0014] R 1 C6~C 14 The aryl group or one or more R 1-1 Substituted C6~C 14 aromatic groups;

[0015] R 1-1 are independently OH, C1-C6 alkyl, C3-C8 cycloalkyl, halogen, NH2, CN, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 alkyl substituted by one or more halogens, C1-C6 alkoxy substituted by one or more halogens, or C1-C6 alkoxy substituted by one or more R 1-1-1 Substituted C3-C8 cycloalkyl;

[0016] R 1-1-1 are independently C1-C6 alkyl;

[0017] R 2 is a 7-12 membered bridged heterocycloalkyl, a 5-8 membered heterocycloalkyl or a 2-1 Substituted 5-8 membered heterocycloalkyl; the heteroatoms in the bridged heterocycloalkyl and heterocycloalkyl groups are independently one or more of nitrogen, oxygen or sulfur, and the number of heteroatoms is independently 1, 2, 3 or 4;

[0018] R 2-1 OH or C1-C6 alkyl;

[0019] X is N or CR 3 ;

[0020] R 3 is a halogen;

[0021] R 4 is a halogen;

[0022] M is a 3- to 10-membered heterocycloalkylene group or a 3- to 10-membered heterocycloalkylene group substituted by one or more halogens; the heteroatoms in the heterocycloalkylene group are each independently one or more of nitrogen, oxygen, or sulfur, and the number of heteroatoms is each independently 1, 2, 3, or 4;

[0023] L is -L1-L2-L3-L4-, L1 is a C1-C6 alkylene group, L2 is a 5-6 membered heterocycloalkylene group, a 7-12 membered spiroheterocycloalkyl group or a 7-12 membered heterocycloalkyl group, L3 is a connecting bond, a C1-C6 alkylene group, an oxygen atom or L4 is a connecting bond or a 5- to 6-membered heterocycloalkylene group; the heteroatoms in the heterocycloalkylene group and the spiroheterocycloalkylene group are each independently one or more of nitrogen, oxygen, or sulfur, and the number of heteroatoms is each independently 1, 2, 3, or 4; wherein L4 is connected to G;

[0024] G is

[0025] Wherein, the compound shown in Formula I is not any of the following compounds:

[0026] In a certain embodiment, the compound as shown in Formula I, or a pharmaceutically acceptable salt thereof:

[0027] Among them, in the compound shown in formula I:

[0028] R 1 C6~C 14 The aryl group or one or more R 1-1 Substituted C6~C 14 aromatic groups;

[0029] R 1-1 are independently OH, C1-C6 alkyl, C3-C8 cycloalkyl, halogen, NH2, CN, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 alkyl substituted by one or more halogens, C1-C6 alkoxy substituted by one or more halogens, or C1-C6 alkoxy substituted by one or more R 1-1-1 Substituted C3-C8 cycloalkyl;

[0030] R 1-1-1 are independently C1-C6 alkyl;

[0031] R 2 is a 7-12 membered bridged heterocycloalkyl, a 5-8 membered heterocycloalkyl, or one or more R 2-1 Substituted 5-8 membered heterocycloalkyl; the heteroatoms in the bridged heterocycloalkyl and heterocycloalkyl groups are independently one or more of nitrogen, oxygen or sulfur, and the number of heteroatoms is independently 1, 2, 3 or 4;

[0032] R 2-1 OH or C1-C6 alkyl;

[0033] X is N or CR 3 ;

[0034] R 3 is a halogen;

[0035] R 4 is a halogen;

[0036] M is a 3- to 10-membered heterocycloalkylene group or a 3- to 10-membered heterocycloalkylene group substituted by one or more halogens; the heteroatoms in the heterocycloalkylene group are each independently one or more of nitrogen, oxygen, or sulfur, and the number of heteroatoms is each independently 1, 2, 3, or 4;

[0037] L is -L1-L2-L3-L4-, L1 is a C1-C6 alkylene group, L2 is a 5-6 membered heterocycloalkylene group or a 7-12 membered spiroheterocycloalkylene group, L3 is a connecting bond, a C1-C6 alkylene group or an oxygen atom, and L4 is a connecting bond or a 5-6 membered heterocycloalkylene group; the heteroatoms in the heterocycloalkylene group and the spiroheterocycloalkylene group are each independently one or more of nitrogen, oxygen or sulfur, and the number of heteroatoms is each independently 1, 2, 3 or 4; wherein L4 is connected to G;

[0038] The above-mentioned connecting bond means that the two groups connected by the bond are directly connected (for example, L is -L1-L2-L3-L4-, and if L3 is a connecting bond, L is -L1-L2-L4-);

[0039] G is

[0040] Wherein, the compound shown in Formula I is not any of the following compounds:

[0041] In one embodiment, the compound of formula I, or a pharmaceutically acceptable salt thereof:

[0042] R 1 C6~C 14 The aryl group or one or more R 1-1 Substituted C6~C 14 aromatic groups;

[0043] R 1-1 are independently OH, C1-C6 alkyl, halogen or C2-C6 alkynyl.

[0044] In one embodiment, in the compound of formula I, or a pharmaceutically acceptable salt thereof, in M, the one or more halogen-substituted 3-10 membered heterocycloalkylene groups are

[0045] The a end is connected to L.

[0046] In one embodiment, the compound of formula I, or a pharmaceutically acceptable salt thereof,

[0047] L is The g end is connected to G.

[0048] In one embodiment, the compound of formula I, or a pharmaceutically acceptable salt thereof,

[0049] L is Wherein the g end is connected to G; preferably

[0050] In one embodiment, the compound of formula I, or a pharmaceutically acceptable salt thereof,

[0051] R 2 for

[0052] In one embodiment, in L2, the heteroatoms in the 7-12 membered heterocycloalkyl group are each independently one or more of nitrogen, oxygen or sulfur, and the number of heteroatoms is each independently 1, 2, 3 or 4.

[0053] In one embodiment, the compound of Formula I, or a pharmaceutically acceptable salt thereof, satisfies one or more of the following conditions:

[0054] (1)R 1 In the above, the C6~C 14 The aryl group and one or more R 1-1 Substituted C6~C 14 The C6~C 14 The aryl groups are independently C6~C 10 The aryl group of

[0055] (2)R 1-1 wherein the C1-C6 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example, ethyl;

[0056] (3)R 1-1 wherein the halogen is independently fluorine, chlorine, bromine or iodine, for example fluorine;

[0057] (4)R 1-1 wherein the C2-C6 alkynyl group is an ethynyl group;

[0058] (5)R 2 In the 7-12 membered bridged heterocycloalkyl, the heteroatom is a nitrogen atom, the number of nitrogen atoms is 2, and one of the nitrogen atoms is connected to the quinazoline ring or the pyridopyrimidine ring; preferably, it is an 8-membered bridged heterocycloalkyl or a 9-membered bridged heterocyclyl, for example: or

[0059] (6)R 2 wherein the 5-8 membered heterocycloalkyl group and one or more R 2-1 In the substituted 5-8 membered heterocycloalkyl, the heteroatom is a nitrogen atom, the number of nitrogen atoms is 1, and the nitrogen atom is connected to the quinazoline ring or the pyridopyrimidine ring; preferably, a 6 membered heterocycloalkyl, for example

[0060] (7)R 2-1 wherein the C1-C6 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example, methyl;

[0061] (8)R 3 、R 4 wherein the halogen is independently fluorine, chlorine, bromine or iodine, for example fluorine;

[0062] (9) In M, the 3- to 10-membered heterocycloalkylene group and the 3- to 10-membered heterocycloalkylene group substituted by one or more halogens are independently 5- to 8-membered monocyclic heterocycloalkylene groups or in wherein Ring A and Ring B are each independently a 3-5 membered saturated heterocyclic ring, wherein the heteroatom in the saturated heterocyclic ring is nitrogen, oxygen or sulfur, and the number of heteroatoms is 1 or 2; Y is C or a heteroatom; preferably, Ring A and Ring B are each independently a 5-membered saturated heterocyclic ring, wherein the heteroatom in the 5-membered saturated heterocyclic ring is, for example, nitrogen, and the number of heteroatoms is, for example, 1; more preferably, Ring A and Ring B are each independently a tetrahydropyrrole ring;

[0063] (10) In M, the halogen in the 3- to 10-membered heterocycloalkylene group substituted by one or more halogens is fluorine, chlorine, bromine or iodine, for example, fluorine;

[0064] (11) In L1 and L3, the C1-C6 alkylene group is independently -CH2-, -CH2CH2-, -CH2CH2CH2-, -CH(CH3)CH2- or -CH2CH(CH3)-, for example, -CH2- or -CH2CH2-;

[0065] (12) In L2, the 5- to 6-membered heterocycloalkylene groups are independently 6-membered heterocycloalkylene groups;

[0066] (13) In L2, the heteroatom of the 5- to 6-membered heterocycloalkylene group is independently nitrogen;

[0067] (14) In L2, the number of heteroatoms in the 5- to 6-membered heterocycloalkylene group is independently 1 or 2;

[0068] (15) In L4, the 5- to 6-membered heterocycloalkylene group is a 6-membered heterocycloalkylene group;

[0069] (16) In L4, the heteroatom of the 5- to 6-membered heterocycloalkylene group is nitrogen;

[0070] (17) In L4, the number of heteroatoms in the 5- to 6-membered heterocycloalkylene group is 2;

[0071] (18) In L2, the 7-12-membered spiroheterocycloalkyl group is an 11-membered spiroheterocycloalkyl group;

[0072] (19) In L2, the heteroatom of the 7-12 membered spiroheterocycloalkyl group is nitrogen;

[0073] (20) In L2, the number of heteroatoms in the 7-12 membered spiroheterocycloalkyl group is 2;

[0074] (21) In L2, the 7-12-membered heterocycloalkyl group is an 8-membered heterocycloalkyl group;

[0075] (22) In L2, the heteroatom of the 7-12 membered heterocycloalkyl group is nitrogen;

[0076] (23) In L2, the number of heteroatoms in the 7-12 membered heterocycloalkyl group is 2.

[0077] In one embodiment, the compound of Formula I or a pharmaceutically acceptable salt thereof satisfies one or more of the following conditions:

[0078] (1) In M, the 3- to 10-membered heterocycloalkylene group and the 3- to 10-membered heterocycloalkylene group substituted by one or more halogens are independently tetrahydropyrrolylene or hexahydro-1H-pyrrolizinylene, for example Wherein end a is connected to L;

[0079] (2) In M, the one or more halogen-substituted 3- to 10-membered heterocycloalkylene groups are Wherein end a is connected to L;

[0080] (3) In L2, the 5- to 6-membered heterocycloalkylene group is a piperidinylene group or a piperazinylene group, for example The d end is connected to L1;

[0081] (4) In L2, the 7-12-membered spiroheterocycloalkyl groups are independently 11-membered spiroheterocyclyl groups, for example The d end is connected to L1;

[0082] (5) In L4, the 5- to 6-membered heterocycloalkylene group is a piperazinylene group or a piperidinylene group, for example The f end is connected to L3;

[0083] (6) In L2, the 7-12-membered cycloheteroalkyl groups are independently 8-membered cycloheterocyclic groups, for example The d end is connected to L1.

[0084] In one embodiment, the compound of Formula I, or a pharmaceutically acceptable salt thereof, satisfies one or more of the following conditions:

[0085] (i): L1 is a C1-C6 alkylene group, L2 is a 5-6 membered heterocycloalkylene group, and L3 and L4 are a connecting bond;

[0086] (ii): L1 is a C1-C6 alkylene group, L2 is a 5-6 membered heterocycloalkylene group, L3 is a C1-C6 alkylene group, and L4 is a 5-6 membered heterocycloalkylene group;

[0087] (iii): L1 is a C1-C6 alkylene group, L2 is a 7-12 membered spiroheterocycloalkyl group, and L3 is L4 is the connecting key;

[0088] (iv): L1 is a C1-C6 alkylene group, L2 is a 5-6 membered heterocycloalkylene group, L3 is an oxygen atom, and L4 is a 5-6 membered heterocycloalkylene group;

[0089] (v): L1 is a C1-C6 alkylene group, L2 is a 7-12 membered spiroheterocycloalkyl group, L3 is a connecting bond, and L4 is a connecting bond;

[0090] (vi): L1 is a C1-C6 alkylene group, L2 is a 7-12 membered heterocycloalkyl group, L3 is a connecting bond, and L4 is a connecting bond.

[0091] In one embodiment, the compound represented by Formula I, or a pharmaceutically acceptable salt thereof, is characterized in that it is represented by the following Formulas I-1, I-2, I-3, I-4 and I-5:

[0092] Among them, R 1 , X, R 4 and M is as defined in any of the above items;

[0093] In formula I-1, G is

[0094] In formula I-2, G is

[0095] In formulas I-3, I-4 and I-5, G is

[0096] In one embodiment, in formula I-4, G is

[0097] In one embodiment, in formula I-5, G is

[0098] In one embodiment, the compound of Formula I, or a pharmaceutically acceptable salt thereof, is characterized in that it satisfies one or more of the following conditions:

[0099] (1) R 1 for

[0100] (2) X is N or CF;

[0101] (3)M is

[0102] (4) L is

[0103] In one embodiment, the compound represented by Formula I, or a pharmaceutically acceptable salt thereof, is characterized in that the compound represented by Formula I is any one of the following compounds:

[0104] Preferably, the pharmaceutically acceptable salt of the compound as shown in Formula I is preferably a formate or hydrochloride salt of the compound as shown in Formula I;

[0105] Preferably, the number of pharmaceutically acceptable salts of the compound represented by Formula I is 1, 2, 3 or 4.

[0106] In one embodiment, the compound represented by Formula I, or a pharmaceutically acceptable salt thereof, is characterized in that the compound represented by Formula I is any one of the following compounds:

[0107] In one embodiment, the compound represented by Formula I, or a pharmaceutically acceptable salt thereof, is characterized in that the compound represented by Formula I is any one of the following compounds:

[0108] The present invention also provides a pharmaceutical composition comprising a substance X and one or more pharmaceutical excipients, wherein the substance X is the compound represented by Formula I, or a pharmaceutically acceptable salt thereof.

[0109] The present invention also provides a use of a substance Y in the preparation of a drug, wherein the drug is a drug for treating or preventing cancer, or a drug for treating or preventing cancer mediated by KRAS mutation;

[0110] In the application, the cancer is preferably blood cancer, pancreatic cancer, MYH-related polyposis, colorectal cancer, lung cancer, non-small cell lung cancer, small cell lung cancer, colorectal cancer, rectal cancer or pancreatic cancer;

[0111] In the application, the KRAS mutation is preferably a KRAS-G12D mutation; the cancer mediated by the KRAS mutation is preferably a blood cancer, pancreatic cancer, MYH-related polyposis, colorectal cancer or lung cancer;

[0112] The substance Y is the compound represented by formula I or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition described above.

[0113] The present invention also provides a method for treating, preventing or treating cancer mediated by KRAS mutation, comprising administering a therapeutically effective amount of substance Y to a patient; the substance Y is the compound shown in Formula I, a pharmaceutically acceptable salt thereof or the pharmaceutical composition described above.

[0114] The cancers mediated by the KRAS mutation include blood cancer, pancreatic cancer, MYH-related polyposis, colorectal cancer or lung cancer.

[0115] The KRAS mutation may be a KRAS_G12D mutation.

[0116] The present invention also provides a method for treating, preventing or treating cancer, comprising administering a therapeutically effective amount of substance Y to a patient; the substance Y is the compound shown in Formula I, a pharmaceutically acceptable salt thereof or the pharmaceutical composition described above; and the cancer is blood cancer, pancreatic cancer, MYH-associated polyposis, colorectal cancer or lung cancer.

[0117] The present disclosure also relates to a method of treating a hyperproliferative disease in a mammal, comprising administering to the mammal a therapeutically effective amount of a compound or a pharmaceutically acceptable salt of the present disclosure.

[0118] Ras mutations, including but not limited to K-Ras, H-Ras or N-Ras mutations, have been identified in hematological cancers or malignancies (e.g., cancers affecting the blood, bone marrow and / or lymph nodes). Accordingly, certain embodiments relate to administering a disclosed compound (e.g., in the form of a pharmaceutical composition) to a patient in need of treatment for a hematological cancer or malignancy.

[0119] In certain specific embodiments, the present disclosure relates to methods for treating lung cancer, comprising administering an effective amount of any of the above-described compounds (or pharmaceutical compositions comprising the compounds) to a subject in need thereof.

[0120] In the present invention, the cancer or malignancy includes, but is not limited to, leukemia and lymphoma. In certain embodiments, the blood disease is further exemplified by acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), chronic myeloid leukemia (CML), acute monocytic leukemia (AMoL), and / or other leukemias. In certain embodiments, the lymphoma is exemplified by all subtypes of Hodgkin's lymphoma or non-Hodgkin's lymphoma.

[0121] In certain embodiments of the present invention, the lung cancer is non-small cell lung cancer (NSCLC), such as adenocarcinoma, squamous cell lung cancer, or large cell lung cancer. In other embodiments, the lung cancer is small cell lung cancer. Other lung cancers include, but are not limited to, adenomas, carcinoids, and undifferentiated carcinomas.

[0122] In some embodiments of the present invention, the cancer, such as acute myeloid leukemia, juvenile cancer, childhood adrenocortical carcinoma, AIDS-related cancers (such as lymphoma and Kaposi's sarcoma), anal cancer, appendix cancer, astrocytoma, atypical malformation, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain stem glioma, brain tumor, breast cancer, bronchial tumor, Burkitt lymphoma, carcinoid, atypical malformation, embryonal tumor, germ cell tumor, primary lymphoma, cervical cancer, childhood cancer, chordoma, heart tumor, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia ( CML), chronic myeloproliferative disorders, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, extrahepatic ductal carcinoma in situ (DCIS), embryonal tumors, central nervous system cancer, endometrial cancer, ependymoma, esophageal cancer, granulomatous neuroblastoma, Ewing's sarcoma, extracranial germ cell tumor, extragonadal germ cell tumor, eye cancer, fibrous histiocytoma of bone, gallbladder cancer, gastric cancer, gastrointestinal carcinoid, gastrointestinal stromal tumor (GIST), germ cell tumor, gestational trophoblastic tumor, hairy cell leukemia, head and neck cancer, heart disease, liver cancer, Hodgkin lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell tumor, pancreatic Neuroendocrine tumors, kidney cancer, laryngeal cancer, lip and oral cavity cancer, liver cancer, lobular carcinoma in situ (LCIS), lung cancer, lymphoma, metastatic squamous cell carcinoma, occult primary, midline cancer, oral cancer, multiple endocrine neoplasia syndrome, multiple myeloma / plasmacytoma, mycosis fungoides, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasms, multiple myeloma, Merkel cell carcinoma, malignant mesothelioma, malignant fibrous histiocytoma of the bone and osteosarcoma, nasal cavity and paranasal sinuses, nasal cavity and paranasal sinus neuroblastoma, non-Hodgkin lymphoma, non-small cell lung cancer (NSCLC), oral cancer, lip and oral cavity cancer , oropharyngeal cancer, ovarian cancer, pancreatic cancer, papillomatosis, paraganglioma, paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pleuropulmonary blastoma, primary central nervous system (CNS) lymphoma, prostate cancer, rectal cancer, transitional cell carcinoma, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, skin cancer, stomach (gastric) cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, T-cell lymphoma, testicular cancer, laryngeal cancer, thymoma and thymic cancer, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, trophoblastic tumor, uncommon childhood cancers, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer, or viral cancer.

[0123] In some embodiments, the non-cancerous hyperproliferative disorder is, for example, a benign hyperplasia of the skin (eg, psoriasis), restenosis, or the prostate (eg, benign prostatic hypertrophy (BPH)).

[0124] Definition of terms

[0125] The term "pharmaceutically acceptable" means that salts, solvents, excipients, etc. are generally non-toxic, safe, and suitable for use by patients. The "patient" is preferably a mammal, more preferably a human.

[0126] The term "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable salt as defined herein and having all the effects of the parent compound. Pharmaceutically acceptable salts can be prepared by adding the corresponding acid in a suitable organic solvent for an organic base and treating according to conventional methods.

[0127] Examples of salt formation include: For base addition salts, it is possible to prepare alkali metal (e.g., sodium, potassium or lithium) or alkaline earth metal (e.g., aluminum, magnesium, calcium, zinc or bismuth) salts by treating a compound of the invention having an appropriate acidic proton with an alkali metal or alkaline earth metal hydroxide or alkoxide (e.g., ethanolate or methanolate) or a suitable basic organic amine (e.g., diethanolamine, choline or meglumine) in an aqueous medium.

[0128] Alternatively, for acid addition salts, salts are formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid; and salts formed with organic acids such as formic acid, acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, glycolic acid, hydroxynaphthoic acid, 2-hydroxyethanesulfonic acid, lactic acid, maleic acid, malic acid, oxalic acid, pyruvic acid, malonic acid, mandelic acid, methanesulfonic acid, mucofuric acid, 2-naphthalenesulfonic acid, propionic acid, salicylic acid, succinic acid, tartaric acid, citric acid, cinnamic acid, p-toluenesulfonic acid or trimethylacetic acid.

[0129] The term "pharmaceutical excipient" may refer to those excipients widely used in the field of pharmaceutical production. Excipients are primarily used to provide a safe, stable, and functional pharmaceutical composition. They may also provide methods to dissolve the active ingredient at a desired rate after administration to a subject, or to promote effective absorption of the active ingredient after administration of the composition to a subject. The pharmaceutical excipients may be inert fillers, or may provide a function, such as stabilizing the overall pH value of the composition or preventing degradation of the active ingredient in the composition. The pharmaceutical excipients may include one or more of the following excipients: binders, suspending agents, emulsifiers, diluents, fillers, granulating agents, adhesives, disintegrants, lubricants, anti-adhesive agents, glidants, wetting agents, gelling agents, absorption delaying agents, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavoring agents, and sweeteners.

[0130] The pharmaceutical compositions of the present invention can be prepared according to the disclosed content using any method known to those skilled in the art, such as conventional mixing, dissolving, granulating, emulsifying, grinding, encapsulating, embedding or lyophilizing processes.

[0131] The pharmaceutical compositions of the present invention can be administered in any form, including injection (intravenous), mucosal, oral (solid and liquid preparations), inhalation, ophthalmic, rectal, topical or parenteral (infusion, injection, implantation, subcutaneous, intravenous, intraarterial, intramuscular) administration. The pharmaceutical compositions of the present invention can also be controlled-release or delayed-release dosage forms (e.g., liposomes or microspheres). Examples of solid oral formulations include, but are not limited to, powders, capsules, caplets, soft capsules, and tablets. Examples of liquid formulations for oral or mucosal administration include, but are not limited to, suspensions, emulsions, elixirs, and solutions. Examples of topical formulations include, but are not limited to, emulsions, gels, ointments, creams, patches, pastes, foams, lotions, drops, or serum preparations. Examples of formulations for parenteral administration include, but are not limited to, solutions for injection, dry preparations that can be dissolved or suspended in a pharmaceutically acceptable carrier, suspensions for injection, and emulsions for injection. Examples of other suitable formulations of the pharmaceutical composition include, but are not limited to, eye drops and other ophthalmic preparations; aerosols such as nasal sprays or inhalers; liquid dosage forms suitable for parenteral administration; suppositories; and lozenges.

[0132] "Treatment" means any treatment of a disease in a mammal, including: (1) preventing the disease, i.e., causing clinical symptoms of the disease not to develop; (2) inhibiting the disease, i.e., preventing the development of clinical symptoms; (3) alleviating the disease, i.e., causing the regression of clinical symptoms.

[0133] "Effective amount" means an amount sufficient, when the compound is administered to a patient in need of treatment, to (i) treat the disease of interest, (ii) reduce, ameliorate, or eliminate one or more symptoms of a particular disease or condition, or (iii) delay the onset of one or more symptoms of a particular disease or condition as described herein. The amount of the carbonyl heterocyclic compound of Formula II or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition described above, corresponding to this amount, will vary depending on factors such as the specific compound, the disease condition and its severity, the characteristics of the patient in need of treatment (e.g., body weight), but can nonetheless be routinely determined by those skilled in the art.

[0134] As used herein, "prevention" refers to a reduction in the risk of acquiring or developing a disease or disorder.

[0135] The term "aryl" refers to a group having a specified number of carbon atoms (e.g., C6 to C 14 ) is a cyclic, unsaturated, monovalent hydrocarbon group, which is monocyclic or polycyclic (for example, 2). When it is polycyclic, the monocyclic rings share two atoms and one bond, and (at least one ring / each ring) has aromaticity, for example, phenyl and naphthyl.

[0136] The term "alkyl" refers to a straight or branched chain alkyl group having the specified number of carbon atoms. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. Unless otherwise specified, an alkyl group is unsubstituted.

[0137] The term "cycloalkyl" refers to a non-aromatic, saturated, monovalent cyclic hydrocarbon group having a specified number of ring carbon atoms (e.g., C3 to C8), which is a single ring. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, and cyclopentyl.

[0138] The term "halogen" refers to F, Cl, Br, I.

[0139] The term "heterocycloalkylene" refers to a non-aromatic saturated bivalent cyclic hydrocarbon group having a specified number of ring atoms (e.g., 3 to 10 members) and at least one ring carbon atom replaced by a heteroatom selected from N, O, and S. Heterocycloalkylene can be connected to other parts of the molecule through a heteroatom or a carbon atom. Examples of monocyclic heterocycloalkylene include, but are not limited to Examples of heterocycloalkylene rings include, but are not limited to,

[0140] In the present invention, the term "alkylene" refers to a saturated, linear or branched divalent hydrocarbon group having a specified number of carbon atoms. Examples of alkylene include, but are not limited to, -CH2-, -CH2CH2-.

[0141] In the present invention, the term "saturated heterocycle" refers to a saturated ring having a specified number of ring atoms (e.g., 3 to 5 members), a specified number of heteroatoms (e.g., 1 or 2), and a specified type of heteroatom (one or more of N, O, and S). Examples of aliphatic heterocycles include, but are not limited to:

[0142] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention.

[0143] Unless otherwise indicated, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of one another.

[0144] Unless otherwise indicated, the term "cis-trans isomers" or "geometric isomers" arises from the inability to rotate freely about double bonds or single bonds forming ring carbon atoms.

[0145] Unless otherwise indicated, the term "diastereomer" refers to stereoisomers that have two or more chiral centers and that are not mirror images of each other.

[0146] Unless otherwise specified, use a solid wedge key. and dotted wedge key To indicate the absolute configuration of a stereocenter, use a straight solid bond and straight dashed key To express the relative configuration of a stereocenter, use The labeled carbon refers to an S-configured chiral carbon, an R-configured chiral carbon, or an achiral carbon.

[0147] Optically active (R)- and (S)-isomers, as well as D and L isomers, can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or base, and then the diastereoisomers are resolved by conventional methods known in the art, and then the pure enantiomer is recovered. In addition, the separation of enantiomers and diastereomers is typically accomplished by using chromatography, which employs a chiral stationary phase and is optionally combined with a chemical derivatization method (e.g., carbamate formation from an amine).

[0148] use It means that the corresponding group is connected to other fragments and groups in the compound through this site.

[0149] On the basis of conforming to the common sense in this field, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0150] The reagents and raw materials used in the present invention are commercially available.

[0151] The positive and progressive effects of the present invention are that the compounds of the present invention have a degradation effect on KRAS protein or a good inhibitory effect on KRAS G12D mutant protein and KRAS mutant protein. DETAILED DESCRIPTION

[0152] The present invention is further illustrated by way of examples below, but the present invention is not limited to the scope of the examples. Experimental methods in the following examples where specific conditions are not specified were performed according to conventional methods and conditions, or selected according to the product specifications.

[0153] In the present invention, the compounds and their salts (or free bases) finally prepared in the following examples, if these compounds have stereoconfigurations generated by axial chirality, the stereoconfigurations of these compounds and their salts (or free bases) generated by axial chirality are consistent with the configurations of the chiral axial intermediates used to prepare these compounds.

[0154] Example 1

[0155] 3-(5-(4-((1-(2-((2S,6R,7aS)-7a-((4-(1R,5S)-3,8-diazabicyclo[3.2.1]oct-3-yl)-7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolidin-2-yl)ethyl)piperidin-4-yl)methyl)piperazin-1-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione

[0156] The synthetic route is as follows:

[0157] Step 1:

[0158] Under 25 degrees Celsius of nitrogen protection stirring conditions, 48-6 (3.0 grams, 6.924 mmoles, 1.00 equivalents), anhydrous tetrahydrofuran (30 milliliters) and hexamethylphosphoric triamide (6 milliliters) were added sequentially in a 100 ml three-necked flask. Under -78 degrees Celsius of nitrogen protection stirring conditions, a tetrahydrofuran solution of lithium diisopropylamide (1 mol per liter, 8.3 milliliters, 8.3 mmoles, 1.2 equivalents) was added dropwise in the reaction solution. The mixture was reacted under 78 degrees Celsius of nitrogen protection stirring conditions for 0.5 hour. Subsequently, allyl bromide (1.06 grams, 8.309 mmoles, 1.2 equivalents) was added dropwise in the reaction solution under 78 degrees Celsius of nitrogen protection stirring conditions. The mixture was slowly raised to 25 degrees Celsius and reacted under 25 degrees Celsius of nitrogen protection stirring conditions for 2 hours. The reaction process was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After the reaction, the reaction mixture was slowly poured into a saturated ammonium chloride solution (100 mL) to quench the reaction. The resulting mixture was extracted with ethyl acetate (100 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The desiccant was filtered to remove the desiccant. The filtrate was evaporated to remove the solvent under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% → 50% methyl tert-butyl ether / petroleum ether. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 53-3 (colorless oil, 1.09 g, 33% yield). MS (ESI, m / z): 452.4 [M+H] + ; 1H NMR (400MHz, CDCl3) δ7.71–7.59(m,4H),7.50–7.36(m,6H),5.77–5.62(m,1H),5.47–5.31(m,1H),5.00–4.88(m,2H),4.21–4.03(m,1H),3.59 –3.51(m,1H),3.42–3.24(m,2H),2.75–2.64(m,1H),2.60–2.43(m,2H) ,2.18–2.06(m,2H),1.97–1.80(m,1H),1.76–1.70(m,1H),1.04(s,9H).

[0159] Step 2:

[0160] Under nitrogen protection and stirring at zero degrees Celsius, a solution of compound 53-3 (600 mg, 1.262 mmol, 1.00 eq) in anhydrous tetrahydrofuran (6 ml) was added dropwise with lithium aluminum tetrahydride in tetrahydrofuran (1 mol / L, 1.9 ml, 1.9 mmol, 1.5 eq). The mixture was reacted at 60 degrees Celsius with nitrogen protection and stirring for 2 hours. The reaction process was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After the reaction was completed, the reaction solution was cooled to zero degrees Celsius, and water (1 ml), sodium hydroxide aqueous solution (15%, 1 ml), and water (3 ml) were added to the reaction solution in sequence. After addition, the mixture was stirred for 15 minutes, and the insoluble matter was removed by filtration. The filter cake was washed with tetrahydrofuran / methanol (3 / 1, 10 ml). The combined filtrate was evaporated to remove the solvent under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% → 10% methanol (1 mol / L ammonia methanol solution) / dichloromethane. The resulting fraction was evaporated to remove the solvent under reduced pressure to afford compound 53-4 (colorless oil, 200 mg, 75% yield). MS (ESI, m / z): 200.1 [M+H] + ; 1 H NMR (400MHz, CDCl3) δ5.84–5.66(m,1H),5.35–5.13(m,1H),5.11–4.88(m,2H),3.40–3.13(m,4H),3.08–2.89(m,1H) ,2.69–2.60(m,1H),2.34–2.19(m,2H),2.18–2.08(m,3H),2.08–1.97(m,1H),1.98–1.88(m,1H),1.68–1.55(m,1H).

[0161] Step 3:

[0162] Under argon protection and stirring at 25 degrees Celsius, compound 53-4 (1.6 g, 8.04 mmol) and N,N-diisopropylethylamine (2.59 g, 20.1 mmol) were added to a solution of compound 1-1 (3.44 g, 8.03 mmol) in anhydrous acetonitrile (20 ml) in sequence. The resulting mixture was reacted at 80 degrees Celsius under argon protection and stirring for 60 hours. The reaction process was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After the reaction was completed, water (25 ml) was added to the reaction solution to quench the reaction. The resulting mixture was extracted with ethyl acetate (25 ml x 3), and the organic phases were combined; the organic phases were washed with saturated brine (30 ml x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography using a 0% → 10% methanol / dichloromethane mobile phase gradient elution. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 1-2 (yellow solid, 3.18 g). MS (ESI, m / z): 591.3 [M+H] + .

[0163] Step 4:

[0164] To a solution of compound 1-2 (3.18 g, 5.38 mmol) in tetrahydrofuran (30 mL) was added potassium phosphate (2.88 g, 13.9 mmol), compound 1-3 (3.48 g, 6.79 mmol), Ad2n-BuP-Pd-G3 (0.99 g, 1.36 mmol), and water (7 mL) in sequence under argon at 25°C. The mixture was stirred at 70°C under argon for 4 hours. The reaction progress was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, the reaction solution was cooled to room temperature and quenched by the addition of water (15 mL). The resulting mixture was extracted with ethyl acetate (15 mL x 3). The organic phases were combined and washed with saturated brine (20 mL x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 1-4 (yellow solid, 4.2 g). MS (ESI, m / z): 941.5 [M+H] + .

[0165] Step 5:

[0166] To a solution of compound 1-4 (2.3 g, 2.44 mmol) in dichloromethane (24 mL) was added potassium osmate dihydrate (0.23 g, 0.62 mmol), N-methylmorpholine N-oxide (1.49 g, 12.7 mmol), and water (6 mL) in sequence under argon at 25°C. The mixture was stirred at room temperature under argon for 16 hours. The reaction process was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, sodium sulfite (15 mL) was added to quench the reaction. The resulting mixture was extracted with dichloromethane (15 mL x 3), and the organic phases were combined; the organic phases were washed with saturated brine (20 mL x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 1-5 (yellow solid, 1.7 g). MS (ESI, m / z): 976.5 [M+H] + .

[0167] Step 6:

[0168] To a solution of compound 1-5 (1.7 g, 1.7 mmol) in N,N-dimethylformamide (15 mL) was added cesium fluoride (0.65 g, 4.3 mmol) at 25°C under argon and stirring. The mixture was stirred at room temperature under argon for 3 hours. The reaction progress was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion, aqueous ammonium chloride (10 mL) was added to quench the reaction. The resulting mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined and washed with saturated brine (10 mL x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% → 10% methanol / dichloromethane. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 1-6 (yellow solid, 0.81 g). MS (ESI, m / z): 819.4 [M+H] + .

[0169] Step 7:

[0170] To a solution of compound 1-6 (169 mg, 0.21 mmol) in acetonitrile (8 mL) was added sodium periodate (88.4 mg, 0.41 mmol) and water (2 mL) at 25°C under argon protection and stirring. The mixture was stirred at room temperature for 15 minutes under argon protection. The reaction process was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After the reaction was completed, aqueous solution (5 mL) was added to quench the reaction. The resulting mixture was extracted with ethyl acetate (5 mL x 3), and the organic phases were combined; the organic phases were washed with saturated brine (5 mL x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain compound 1-7 (yellow solid, 158 mg). MS (ESI, m / z): 787.7 [M+H] + .

[0171] Step 8:

[0172] To a solution of compound 1-7 (110 mg, 0.14 mmol) in methanol (5 mL) was added compound 1-8 (146 mg, 0.34 mmol) and sodium acetate (60 mg, 0.73 mmol) at 25°C under argon and stirring. The mixture was stirred at room temperature for 15 minutes under argon, after which sodium cyanoborohydride (28 mg, 0.44 mmol) was added. The mixture was stirred at room temperature under argon for 16 hours. The reaction progress was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, aqueous solution (5 mL) was added to quench the reaction. The resulting mixture was extracted with ethyl acetate (5 mL x 3), and the organic phases were combined; the organic phases were washed with saturated brine (5 mL x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 1-9 (yellow solid, 103 mg). MS (ESI, m / z): 1197.4 [M+H] + .

[0173] Step 9:

[0174] To a solution of compound 1-9 (103 mg, 0.086 mmol) in acetonitrile (2 mL) was added a solution of dioxane hydrochloride (4 mol / L, 2 mL) at 25°C with stirring under argon. The mixture was stirred at room temperature for 2 hours under argon. The reaction progress was monitored by LC / MS and thin-layer chromatography. After completion of the reaction, the mixture was purified by HPLC using an XBridge Prep OBD C18 Column, 30 x 150 mm, 5 μm; mobile phase A: water (10 mmol / L formic acid), mobile phase B: acetonitrile; flow rate: 60 mL / min; elution was performed with 5% → 35% phase B over 12 minutes; UV detection: 220 / 254 nm. This yielded the formate salt of compound 1 (52 mg, white solid). 1 H NMR (400MHz, Methanol-d4) δ9.04(s,1H),8.50(s,2H),7.87(dd,J=9.2,5.7Hz,1H),7.64(d,J=8.4Hz,1H),7.40–7. 29(m,2H),7.22(d,J=2.5Hz,1H),7.07(d,J=9.9Hz,2H),5.44–5.26(m,2H),5.12–5.07(m,1H),4.68–4.55(m,5H),4. 46–4.34(m,3H),4.25(dd,J=10.7,7.8Hz,1H),3.91–3.74(m,4H),3.58–3.37(m,4H),3.27–2.99(m,4H),2.96–2.75( m,5H),2.57–2.37(m,6H),2.35–2.09(m,6H),2.00–1.74(m,8H),1.43(d,J=12.7Hz,2H).MS(ESI,m / z):1053.2[M+H] + .

[0175] Example 2

[0176] 3-(5-(4-((1-(2-((2S,6R,7aS)-7a-(((4-(3,9-diazabicyclo[4.2.1]non-3-yl))-7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolidin-2-yl)ethyl)piperidin-4-yl)methyl)piperazin-1-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione

[0177] The synthetic route is as follows:

[0178] Step 1:

[0179] Compound 2-1 (1 g, 2.26 mmol), compound 53-4 (450 mg, 2.26 mmol), and DIEA (731 mg, 5.65 mmol) were dissolved in acetonitrile (10 mL) under nitrogen and stirred at 25°C. The mixture was stirred at 80°C for 72 hours. The reaction solution was concentrated, and the crude product was purified by column chromatography (ethyl acetate / petroleum ether = 0-50%, then methanol / dichloromethane = 0-10%) to obtain compound 2-2 (colorless oil, 980 mg, 71.63% yield). MS (ESI, m / z): 605.0 [M+H]+.

[0180] Step 2:

[0181] Compound 2-2 (200 mg, 0.33 mmol), compound 1-2 (203 mg, 0.10 mmol), potassium phosphate (210 mg, 0.99 mmol), and Ad2nBuP-Pd-G3 (24 mg, 0.033 mmol) were dissolved in tetrahydrofuran (2 mL) and water (0.1 mL) under nitrogen at 25°C. The reaction mixture was stirred at 90°C under nitrogen for 16 hours. The reaction mixture was filtered and concentrated, and the crude product was purified by column chromatography (methanol / dichloromethane = 0-10%) to obtain compound 2-3 (yellow solid, 200 mg, 63.5% yield). MS (ESI, m / z): 955.0 [M+H]+.

[0182] Step 3:

[0183] Compound 2-3 (200 mg, 0.21 mmol) and potassium osmate dihydrate (6.5 mg, 0.02 mmol) were dissolved in dichloromethane (8 mL) and water (2 mL) under nitrogen and stirred at 25°C. The reaction mixture was stirred for 16 hours and then extracted with dichloromethane (10 mL x 2). The organic phase was filtered and concentrated, and the crude product was purified by preparative TLC (dichloromethane / methanol = 10:1) to obtain compound 2-4 (yellow solid, 85 mg, 41% yield). MS (ESI, m / z): 988.0 [M+H]+.

[0184] Step 4:

[0185] Compound 2-4 (85 mg, 0.086 mmol) was dissolved in acetonitrile (8 mL) and water (8 mL) under nitrogen and stirred at 25°C. Sodium periodate (55 mg, 0.26 mmol) was added. The reaction mixture was stirred at 25°C for 1 hour, diluted with water (10 mL), and extracted with ethyl acetate (10 mL x 2). The organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by preparative TLC (dichloromethane / methanol = 10:1) to obtain compound 2-5 (yellow solid, 80 mg, 97.26% yield). MS (ESI, m / z): 957.0 [M+H]+.

[0186] Step 5:

[0187] Compound 2-6 (108 mg, 0.046 mmol) was dissolved in methanol (10 mL), and sodium acetate (55 mg, 0.66 mmol) was added. The reaction mixture was stirred at 20°C for 10 minutes. Compound 2-5 (80 mg, 0.083 mmol) was then added, and the reaction mixture was stirred at 20°C for 30 minutes. Sodium cyanoborohydride (16 mg, 0.25 mmol) was then added, and the reaction mixture was stirred at 20°C for 16 hours. The reaction mixture was diluted with dichloromethane (30 mL), washed with water (10 mL x 2) and saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting crude product was purified on a preparative plate (dichloromethane:methanol = 10:1) to afford compound 2-7 (white solid, 90 mg, 78.79% yield). MS (ESI, m / z): 1366.0 [M+H]+.

[0188] Step 6:

[0189] Compound 2-7 (80 mg, 0.059 mmol) was dissolved in DMF (1 mL) under nitrogen and stirred at 25°C. CsF (89 mg, 0.58 mmol) was added. The reaction mixture was stirred at 25°C for 1 hour. The reaction mixture was concentrated, and the crude product was purified by preparative TLC (dichloromethane / methanol = 10:1) to obtain compound 2-8 (yellow solid, 50 mg, 70.57% yield). MS (ESI, m / z): 1210.0 [M+H]+.

[0190] Step 7:

[0191] Compound 2-8 (50 mg, 0.041 mmol) was dissolved in acetonitrile (2 mL) at 0°C, and a 4 M hydrochloric acid solution in dioxane (2 mL) was added. The reaction mixture was stirred at 20°C for 1 hour. The crude product obtained by concentration was purified by preparative HPLC to obtain the formate salt of compound 2 (yellow solid, 23 mg, 53.29% yield). MS (ESI, m / z): 1066.0 [M+H]+. 1H NMR(400MHz,MeOD)9.09(d,J=9.9Hz,1H),8.49(s,1H),7.95–7.84(m,1H),7.66(d,J=8.4Hz,1H ),7.36(dd,J=17.2,5.6Hz,2H),7.25(s,1H),7.09(d,J=10.3Hz,2H),5.37(d,J=52.2Hz,1H),5. 12(dd,J=13.4,5.1Hz,1H),4.73(d,J=12.5Hz,1H),4.54–4.21(m,5H),4.03(dd,J=47.8,29.2Hz ,4H),3.34(s,8H),3.32–3.01(m,5H),2.99–2.72(m,5H),2.57(s,23H),1.45(d,J=10.9Hz,2H).

[0192] Example 3

[0193] 5-(4-(4-(2-((2S,6R,7aS)-7a-((4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-6,8-difluoroquinazolin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolizin-2-yl)ethyl)piperazin-1-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0194] The synthetic route is as follows:

[0195] Step 1:

[0196] To a reaction flask, 53-4 (200 mg, 0.952 mmol, 1.00 equiv), compound 46-1 (645.42 mg, 0.952 mmol, 1.00 equiv), cesium carbonate (654.04 mg, 1.904 mmol, 2.00 equiv), triethylenediamine (11.24 mg, 0.096 mmol, 0.10 equiv), and N,N-dimethylformamide (6 mL) were added sequentially under nitrogen protection and stirring at 25°C. The mixture was stirred at 80°C under nitrogen protection for 2 hours. The reaction progress was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, the reaction solution was poured into ice water (60 mL) for quenching, and the resulting mixture was extracted with ethyl acetate (60 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was evaporated to remove the solvent under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography using a 0% → 10% methanol / dichloromethane mobile phase gradient elution. The resulting fractions were evaporated to remove the solvent under reduced pressure to obtain compound 53-5 (white solid, 460.0 mg, 56% yield). MS (ESI, m / z): 806.3 [M+H] + .

[0197] Step 2:

[0198] Compound 53-5 (460 mg) obtained in step 5 of this example was subjected to chiral separation by supercritical fluid chromatography: chiral column NB_CHIRALPAK AD, 3 x 25 cm, 5 μm; mobile phase A: supercritical carbon dioxide, mobile phase B: isopropanol (0.1% 2 mol / L ammonia in methanol); flow rate: 100 ml / min; elution was performed with 50% mobile phase B; detector: UV 220 / 206 nm, yielding two products. The product with the shorter retention time (1.68 minutes) was compound 53-5a (white solid, 200 mg, 45% recovery); MS (ESI, m / z): 806.4 [M+H] + The product with a longer retention time (3.63 minutes) was compound 53-5b (white solid, 220 mg, recovery rate 48%), MS (ESI, m / z): 806.4 [M+H] + .

[0199] Step 3:

[0200] To a solution of compound 53-5a (1.1 g, 1.3 mmol) in dichloromethane (12 mL) was added potassium osmate dihydrate (60 mg, 0.16 mmol), N-methylmorpholine N-oxide (300 mg, 2.56 mmol), and water (3 mL) in sequence under argon at 25°C. The mixture was stirred at room temperature under argon for 2 hours. The reaction progress was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, sodium sulfite (15 mL) was added to quench the reaction. The resulting mixture was extracted with dichloromethane (15 mL x 3), and the organic phases were combined; the organic phases were washed with saturated brine (20 mL x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane. The resulting fraction was evaporated to remove the solvent under reduced pressure to afford compound 53-6a (pale yellow solid, 1.1 g). MS (ESI, m / z): 840.9 [M+H] + .

[0201] Step 4:

[0202] To a solution of compound 53-6a (400 mg, 0.48 mmol) in acetonitrile (16 ml) was added sodium periodate (204 mg, 0.41 mmol) and water (4 ml) at 25 degrees Celsius under argon protection and stirring. The mixture was stirred at room temperature under argon protection for 15 minutes. The reaction process was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After the reaction was completed, aqueous solution (8 ml) was added to quench the reaction. The resulting mixture was extracted with ethyl acetate (8 ml x 3), and the organic phases were combined; the organic phases were washed with saturated brine (8 ml x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain compound 53-7a (pale yellow solid, 375 mg). MS (ESI, m / z): 808.9 [M+H] + .

[0203] Step 5:

[0204] To a solution of compound 53-7a (96 mg, 0.12 mmol) in methanol (10 mL) was added compound 3-1 (105 mg, 0.24 mmol) and sodium acetate (49 mg, 0.60 mmol) at 25°C under argon and stirring. The mixture was stirred at room temperature for 15 minutes under argon, after which sodium cyanoborohydride (22 mg, 0.36 mmol) was added. The mixture was stirred at room temperature under argon for 16 hours. The reaction progress was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, aqueous solution (5 mL) was added to quench the reaction. The resulting mixture was extracted with ethyl acetate (5 mL x 3), and the organic phases were combined; the organic phases were washed with saturated brine (5 mL x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 3-2 (pale yellow solid, 132 mg). MS (ESI, m / z): 1232.4 [M+H] + .

[0205] Step 6:

[0206] To a solution of compound 3-2 (127 mg, 0.10 mmol) in dichloromethane (3 mL) was added trifluoroacetic acid (3 mL) under stirring at 25°C under argon. The mixture was stirred at room temperature for 2 hours under argon. The reaction progress was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, the mixture was purified by HPLC using an XBridge Prep OBD C18 Column, 30 x 150 mm, 5 μm; mobile phase A: water (10 mmol / L formic acid), mobile phase B: acetonitrile; flow rate: 60 mL / min; elution was performed with 5% → 35% phase B over 12 minutes; UV detection: 220 / 254 nm. This yielded the formate salt of compound 3 (yellow-green solid, 37.2 mg). 1H NMR(400MHz, Methanol-d4)δ8.49(s,1H),7.73–7.58(m,3H),7.36–7.16(m,4H),6.98(d,J=2.6Hz,1H), 5.46–5.31(m,1H),5.06(dd,J=12.4,5.5Hz,1H),4.59–4.50(m,4H),4.40–4.29(m,2H),4.05–3.92(m,4H ),3.77–3.68(m,2H),3.54–3.34(m,3H),3.01–2.93(m,2H),2.87–2.56(m,13H),2.50–2.26(m,7H),2.1 2–2.00(m,5H),1.92–1.67(m,6H),1.36–1.24(m,4H),0.80(t,J=7.4Hz,3H).MS(ESI,m / z):1088.2[M+H] + .

[0207] Example 4

[0208] 5-(4-(4-(2-((2S,6R,7aS)-7a-((4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-6,8-difluoroquinazolin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolizin-2-yl)ethyl)piperazin-1-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)-6-fluoroisoindoline-1,3-dione

[0209] The synthetic route is as follows:

[0210] Step 1:

[0211] To a solution of compound 53-7a (100 mg, 0.12 mmol) in methanol (10 mL) was added compound 4-1 (110 mg, 0.24 mmol) and sodium acetate (49 mg, 0.60 mmol) at 25°C with stirring under argon. The mixture was stirred at room temperature for 15 minutes under argon, after which sodium cyanoborohydride (22 mg, 0.36 mmol) was added. The mixture was stirred at room temperature under argon for 16 hours. The reaction progress was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, aqueous solution (5 mL) was added to quench the reaction. The resulting mixture was extracted with ethyl acetate (5 mL x 3), and the organic phases were combined; the organic phases were washed with saturated brine (5 mL x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 4-2 (pale yellow solid, 149 mg). MS (ESI, m / z): 1250.4 [M+H] + .

[0212] Step 2:

[0213] To a solution of compound 4-2 (144 mg, 0.12 mmol) in dichloromethane (3 mL) was added trifluoroacetic acid (3 mL) under stirring at 25°C under argon. The mixture was stirred at room temperature under argon for 2 hours. The reaction progress was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, the mixture was purified by HPLC using an XBridge Prep OBD C18 Column, 30 x 150 mm, 5 μm; mobile phase A: water (10 mmol / L formic acid), mobile phase B: acetonitrile; flow rate: 60 mL / min; elution was performed with 5% → 35% phase B over 12 minutes; UV detection: 220 / 254 nm. This yielded the formate salt of compound 4 (yellow-green solid, 48.7 mg). 1H NMR (400MHz, Methanol-d4) δ8.52(s,1H),7.77–7.63(m,2H),7.51(dd,J=25.9,9.2Hz,2H),7.37–7.21(m,2H),7.00(d,J=2.6Hz,1H ),5.48–5.33(m,1H),5.10(dd,J=12.5,5.4Hz,1H),4.57(dd,J=20.1,13.4Hz,2H),4.45–4.31(m,2H),3.99(s,2H),3.80–3.58(m,4H ),3.52(t,J=7.8Hz,1H),3.40(dd,J=11.3,8.5Hz,2H),3.28(d,J=16.2Hz,1H),2.97–2.77(m,8H),2.77–2.51(m,8H),2.51–2.19(m ,7H),2.18–2.00(m,5H),1.95–1.65(m,6H),1.37(ddd,J=26.0,15.0,5.9Hz,3H),0.82(t,J=7.4Hz,3H).MS(ESI,m / z):1106.2[M+H] + .

[0214] Example 5

[0215] 5-(4-((1-(2-((2S,6R,7aS)-7a-((((S))-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octane)-3-yl)-7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-6,8-difluoroquinazolin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolizin-2-yl)ethyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0216] The synthetic route is as follows:

[0217] Step 1:

[0218] Compound 5-1 (65 mg, 0.15 mmol) was dissolved in methanol (10 mL), and sodium acetate (81 mg, 0.99 mmol) was added. The reaction solution was stirred at 20°C for 10 minutes. Compound 53-7a (100 mg, 0.12 mmol) was added, and the reaction solution was stirred at 20°C for 30 minutes. Sodium cyanoborohydride (23 mg, 0.37 mmol) was added, and the reaction solution was stirred at 20°C for 3 hours. The reaction solution was diluted with dichloromethane (30 mL), washed with water (10 mL x 2) and saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on a preparative plate (dichloromethane:methanol = 0-10%) to obtain compound 5-2 (yellow solid, 90 mg, 59.10% yield). MS (ESI, m / z): 1230.0 [M+H]+.

[0219] Step 2:

[0220] Compound 5-2 (80 mg, 0.065 mmol) was dissolved in dichloromethane (1 mL), and trifluoroacetic acid (1 mL) was added at 20°C. The reaction mixture was stirred at 20°C for 1 hour, concentrated, and the resulting crude product was purified by preparative HPLC to give the formate salt of compound 5 (red solid, 10 mg, 14.16% yield). MS (ESI, m / z): 1086.0 [M+H]+. 1H NMR(400MHz,MeOD)δ8.55(s,1H),7.73–7.62(m,2H),7.41–7.13(m,4H),7.00(d,J=2.4Hz,1H),5.36(d ,J=52.9Hz,2H),5.08(dd,J=12.5,5.5Hz,1H),4.51(dd,J=24.7,12.2Hz,2H),4.34(d,J=10.6Hz,1H), 4.22(d,J=10.6Hz,1H),4.03(d,J=13.8Hz,1H),3.80–3.57(m,4H),3.44(d,J=47.6Hz,3H),3.20(d,J= 40.8Hz,3H),3.03–2.08(m,18H),1.99–1.63(m,10H),1.46–1.05(m,7H),0.84(dd,J=16.7,9.4Hz,3H).

[0221] Example 6

[0222] 5-(4-((1-(2-((2S,6R,7aS)-7a-((((7S)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl))-7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-6,8-difluoroquinazolin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolizin-2-yl)ethyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)-6-fluoroisoindoline-1,3-dione

[0223] The synthetic route is as follows:

[0224] Step 1:

[0225] Compound 6-1 (65 mg, 0.15 mmol) was dissolved in methanol (10 mL), and sodium acetate (81 mg, 0.99 mmol) was added. The reaction solution was stirred at 20°C for 10 minutes. Compound 53-7a (100 mg, 0.12 mmol) was added, and the reaction solution was stirred at 20°C for 30 minutes. Sodium cyanoborohydride (23 mg, 0.37 mmol) was added, and the reaction solution was stirred at 20°C for 3 hours. The reaction solution was diluted with dichloromethane (30 mL), washed with water (10 mL x 2) and saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on a preparative plate (dichloromethane:methanol = 0-10%) to obtain compound 6-2 (yellow solid, 90 mg, 59.10% yield). MS (ESI, m / z): 1249.0 [M+H]+.

[0226] Step 2:

[0227] Compound 6-2 (80 mg, 0.065 mmol) was dissolved in dichloromethane (1 mL), and trifluoroacetic acid (1 mL) was added at 20°C. The reaction mixture was stirred at 20°C for 1 hour, concentrated, and the resulting crude product was purified by preparative HPLC to afford the formate salt of compound 6 (red solid, 35 mg, 49.46% yield). MS (ESI, m / z): 552.9 [M / 2+H]+. 1HNMR(400MHz,MeOD)δ8.53(s,1H),7.73–7.63(m,3H),7.54(d,J=11.3Hz,1H),7.47(d,J=7.4Hz,1H),7.34–7.24(m,2H),6.99 (d,J=2.6Hz,1H),5.43(s,1H),5.30(s,1H),5.10(dd,J=12.3,5.7Hz,1H),4.52(dd,J=25.2,13.2Hz,3H),4.34(d,J=10.8Hz,1H ),4.23(d,J=10.3Hz,1H),3.80–3.62(m,5H),3.50(s,3H),3.15(s,3H),3.00(s,2H),2.93–2.83(m,3H),2.82–2.71(m,4H),2. 38(dd,J=50.5,15.1Hz,5H),2.15(s,3H),1.83(dd,J=46.7,36.6Hz,10H),1.35(dd,J=41.9,9.2Hz,7H),0.82(t,J=7.4Hz,3H).

[0228] Example 7

[0229] 5-(4-((1-(2-((2S,6R,7aS)-7a-((4-(1R,5S)-3,8-diazabicyclo[3.2.1]oct-3-yl)-7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolidin-2-yl)ethyl)piperidin-4-yl)methyl)-6-fluorohexahydro-1H-pyrrolidin-2-yl)ethyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0230] The synthetic route is as follows:

[0231] Step 1:

[0232] To a solution of compound 1-7 (79 mg, 0.10 mmol) in methanol (5 mL) was added compound 7-1 (90 mg, 0.21 mmol) and sodium acetate (42 mg, 0.52 mmol) at 25°C under argon protection and stirring. The mixture was stirred at room temperature for 15 minutes under argon protection. Then, sodium cyanoborohydride (19 mg, 0.31 mmol) was added to the mixture. The mixture was stirred at room temperature under argon protection for 16 hours. The reaction process was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, aqueous solution (5 mL) was added to quench the reaction. The resulting mixture was extracted with ethyl acetate (5 mL x 3), and the organic phases were combined; the organic phases were washed with saturated brine (5 mL x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 7-2 (yellow solid, 52 mg). MS (ESI, m / z): 1210.4 [M+H] + .

[0233] Step 2:

[0234] To a solution of compound 7-2 (52 mg, 0.043 mmol) in acetonitrile (2 mL) was added a solution of dioxane hydrochloride (4 mol / L, 2 mL) under argon at 25°C. The mixture was stirred at room temperature under argon for 2 hours. The reaction progress was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, the mixture was purified by HPLC using an XBridge Prep OBD C18 Column, 30 x 150 mm, 5 μm; mobile phase A: water (10 mmol / L formic acid), mobile phase B: acetonitrile; flow rate: 60 mL / min; elution was performed with 5% → 35% phase B over 12 minutes; UV detection: 220 / 254 nm. This yielded the formate salt of compound 7 (yellow-green solid, 24.8 mg). 1H NMR(400MHz, Methanol-d4)δ9.02(s,1H),7.86(dd,J=9.1,5.7Hz,1H),7.65(dd,J=8.6,0.9Hz,1H),7.37–7.29(m,3H),7.24–7.1 5(m,2H),5.40(d,J=3.7Hz,1H),5.06(dd,J=12.5,5.4Hz,2H),4.62(d,J=14.7Hz,7H),4.33(t,J=11.0Hz,1H),4.21(dd,J=10.6,7 .4Hz,1H),4.00(d,J=12.9Hz,2H),3.75–3.65(m,4H),3.35(dd,J=2.1,1.0Hz,3H),3.28–3.10(m,3H),2.95(t,J=12.8Hz,2H),2.8 7–2.70(m,6H),2.53(t,J=12.2Hz,2H),2.39–2.06(m,5H),1.86–1.70(m,9H),1.31(d,J=18.9Hz,5H).MS(ESI,m / z):1066.2[M+H] + .

[0235] Example 8

[0236] 5-(4-((1-(2-((2S,6R,7aS)-7a-((4-(1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolazin-2-yl)ethyl)piperidin-4-yl)methyl)-6-fluorohexahydro-1H-pyrrolazin-2-yl)ethyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)-6-fluoroisoindoline-1,3-dione

[0237] The synthetic route is as follows:

[0238] Step 1:

[0239] To a solution of compound 1-7 (79 mg, 0.10 mmol) in methanol (5 mL) was added compound 8-1 (94 mg, 0.21 mmol) and sodium acetate (42 mg, 0.52 mmol) at 25°C under argon and stirring. The mixture was stirred at room temperature for 15 minutes under argon, after which sodium cyanoborohydride (19 mg, 0.31 mmol) was added. The mixture was stirred at room temperature under argon for 16 hours. The reaction progress was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, aqueous solution (5 mL) was added to quench the reaction. The resulting mixture was extracted with ethyl acetate (5 mL x 3), and the organic phases were combined; the organic phases were washed with saturated brine (5 mL x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 8-2 (yellow solid, 50 mg). MS (ESI, m / z): 1228.4 [M+H] + .

[0240] Step 2:

[0241] To a solution of compound 8-2 (50 mg, 0.041 mmol) in acetonitrile (2 mL) was added a solution of dioxane hydrochloride (4 mol / L, 2 mL) under argon and stirred at 25°C. The mixture was stirred at room temperature under argon for 2 hours. The reaction progress was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, the mixture was purified by HPLC using an XBridge Prep OBD C18 Column, 30 x 150 mm, 5 μm; mobile phase A: water (10 mmol / L formic acid), mobile phase B: acetonitrile; flow rate: 60 mL / min; elution was performed with 5% → 35% phase B over 12 minutes; UV detection: 220 / 254 nm. This yielded the hydrochloride salt of compound 8 (yellow-green solid, 14.3 mg). 1H NMR(400MHz, Methanol-d4)δ9.02(s,1H),7.87(dd,J=9.2,5.7Hz,1H),7.53–7.43(m,2H),7.37–7.3 0(m,2H),7.23–7.19(m,1H),5.39–5.26(m,1H),5.08(dd,J=12.5,5.4Hz,2H),4.33(dd,J=13.8,10. 6Hz,1H),4.25–4.16(m,1H),3.75–3.61(m,7H),3.27–3.07(m,5H),2.96–2.55(m,10H),2.43–2.06( m,9H),1.80(d,J=9.8Hz,7H),1.71–1.58(m,4H),1.24(d,J=6.2Hz,4H).MS(ESI,m / z):1084.2[M+H] + .

[0242] Example 9

[0243] 3-(5-(4-((1-(2-((2S,6R,7aS)-7a-((((S))-4-((1R,5S)-3,8-diazabicyclo)[3.2.1]oct-3-yl)-7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-6,8-difluoroquinazolin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolizin-2-yl)ethyl)piperidin-4-yl)methyl)piperidin-1-yl)-6-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione

[0244] The synthetic route is as follows:

[0245] Step 1:

[0246] Compound 9-1 (120 mg, 0.22 mmol) was dissolved in dichloromethane (10 mL), and a dioxane hydrochloride solution (4 M, 2 mL, 8 mmol) was added. The reaction mixture was stirred at 20°C for 30 minutes. The reaction mixture was concentrated to give compound 9-2 (white solid, 100 mg, 100% yield). MS (ESI, m / z): 443.0 [M+H]+.

[0247] Step 2:

[0248] Compound 9-2 (77 mg, 0.17 mmol) was dissolved in methanol (10 mL), and sodium acetate (114 mg, 1.38 mmol) was added. The reaction solution was stirred at 20°C for 10 minutes. Compound 53-7a (140 mg, 0.17 mmol) was added, and the reaction solution was stirred at 20°C for 30 minutes. Sodium cyanoborohydride (33 mg, 0.52 mmol) was added, and the reaction solution was stirred at 20°C for 16 hours. The reaction solution was diluted with dichloromethane (30 mL), washed with water (10 mL x 2) and saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on a preparative plate (dichloromethane:methanol = 0-10%) to obtain compound 9-3 (white solid, 120 mg, 56.10% yield). MS (ESI, m / z): 1234.0 [M+H]+.

[0249] Step 3:

[0250] Compound 9-3 (115 mg, 0.093 mmol) was dissolved in dichloromethane (1 mL), and trifluoroacetic acid (1 mL) was added at 20°C. The reaction solution was stirred at 20°C for 1 hour, and the reaction solution was concentrated. The crude product was purified by preparative HPLC to give the formate salt of compound 9 (white solid, 5 mg, 4.92% yield). MS (ESI, m / z): 1090.0 [M+H]+. 1H NMR(400MHz,MeOD)δ8.53(s,1H),7.75–7.62(m,2H),7.42(d,J=11.5Hz,1H),7.27(ddd,J=34.2,17.7,7.6Hz,3H),7 .00(d,J=2.5Hz,1H),5.37(d,J=52.1Hz,1H),5.12(dd,J=13.4,5.1Hz,1H),4.56(dd,J=25.5,12.9Hz,2H),4.39(dd, J=22.3,8.4Hz,2H),4.24(d,J=10.7Hz,1H),3.95(s,2H),3.73(dd,J=21.6,13.3Hz,2H),3.62–3.41(m,5H),3.29–2 .95(m,4H),2.94–2.09(m,16H),2.09–1.65(m,12H),1.57(s,1H),1.48–1.22(m,6H),0.83(dd,J=17.8,10.5Hz,3H).

[0251] Example 10

[0252] 3-(5-(4-((4-(2-((2S,6R,7aS)-7a-((((S))-4-((1R,5S)-3,8-diazabicyclo)[3.2.1]oct-3-yl)-7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-6,8-difluoroquinazolin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolizin-2-yl)ethyl)piperazin-1-yl)methyl)piperidin-1-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione

[0253] The synthetic route is as follows:

[0254] Step 1:

[0255] Compound 10-1 (175 mg, 0.62 mmol) and compound 10-2 (200 mg, 0.62 mmol) were dissolved in dioxane (4 mL), and palladium acetate (28 mg, 0.12 mmol), X-phos (118 mg, 0.25 mmol), and cesium carbonate (403 mg, 1.24 mmol) were added sequentially. The reaction mixture was stirred at 100°C under nitrogen for 16 hours. The reaction mixture was filtered and concentrated, and the crude product was purified by column chromatography (methanol / dichloromethane = 0-8%) to obtain compound 10-3 (brown solid, 190 mg, 58.51% yield). MS (ESI, m / z): 425.0 [M-100+H]+.

[0256] Step 2:

[0257] Compound 10-3 (190 mg, 0.36 mmol) was dissolved in dichloromethane (2 mL), and a dioxane hydrochloride solution (4 M, 2 mL, 8 mmol) was added. The reaction mixture was stirred at 20°C for 1 hour. The reaction mixture was concentrated to give compound 10-4 (brown solid, 150 mg, 97.56% yield). MS (ESI, m / z): 426.0 [M+H]+.

[0258] Step 3:

[0259] Compound 10-4 (79 mg, 0.19 mmol) was dissolved in methanol (10 mL), and sodium acetate (122 mg, 1.49 mmol) was added. The reaction solution was stirred at 20°C for 10 minutes. Compound 53-7a (150 mg, 0.19 mmol) was added, and the reaction solution was stirred at 20°C for 30 minutes. Sodium cyanoborohydride (35 mg, 0.56 mmol) was added, and the reaction solution was stirred at 20°C for 16 hours. The reaction solution was diluted with dichloromethane (30 mL), washed with water (10 mL x 2) and saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on a preparative plate (dichloromethane:methanol = 0-10%) to obtain compound 10-5 (white solid, 60 mg, 26.57% yield). MS (ESI, m / z): 1217.0 [M+H]+.

[0260] Step 4:

[0261] Compound 10-5 (50 mg, 0.041 mmol) was dissolved in acetonitrile (1 mL) and a dioxane hydrochloride solution (4 M, 1 mL, 4 mmol) was added at 20°C. The reaction mixture was stirred at 20°C for 1 hour, concentrated, and the resulting crude product was purified by preparative HPLC to afford the formate salt of compound 10 (white solid, 23 mg, 52.18% yield). MS (ESI, m / z): 537.0 [M / 2+H]+. 1H NMR (400MHz, MeOD) δ8.54(s,1H),7.76–7.53(m,3H),7.35–7.21(m,2H),7.07(d,J=8.9Hz,2H),7.00(d,J=2.5Hz,1H),5.43(d,1H),5.13 –5.09(m,1H),4.55(dd,J=26.4,12.9Hz,3H),4.42–4.33(m,3H),4.24(d,J=10.5Hz,1H),3.90(s,4H),3.72(dd,J=22.1,12.8Hz,2H),3. 43(d,J=12.9Hz,3H),3.23(d,J=41.0Hz,2H),3.04(s,2H),2.94–2.79(m,6H),2.64–2.55(m,1H),2.51–2.39(m,4H),2.32(d,J=21.8Hz, 2H),2.25–2.14(m,2H),2.05–1.93(m,6H),1.87–1.70(m,6H),1.41(dd,J=28.4,15.3Hz,5H),1.28–1.20(m,2H),0.82(t,J=7.4Hz,3H).

[0262] Example 11

[0263] 5-(9-(2-((2S,6R,7aS)-7a-((((7S)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl))-7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-6,8-difluoroquinazolin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolizin-2-yl)ethyl)-3,9-diazaspiro[5.5]undec-3-yl)-2-(2,6-dioxopiperidin-3-yl)-6-fluoroisoindoline-1,3-dione

[0264] The synthetic route is as follows:

[0265] Step 1:

[0266] Compound 11-1 (80 mg, 0.18 mmol) was dissolved in methanol (10 mL), and sodium acetate (122 mg, 1.49 mmol) was added. The reaction solution was stirred at 20°C for 10 minutes. Compound 53-7a (150 mg, 0.19 mmol) was added, and the reaction solution was stirred at 20°C for 30 minutes. Sodium cyanoborohydride (35 mg, 0.56 mmol) was added, and the reaction solution was stirred at 20°C for 16 hours. The reaction solution was diluted with dichloromethane (30 mL), washed with water (10 mL x 2) and saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on a preparative plate (dichloromethane:methanol = 0-10%) to obtain compound 11-2 (white solid, 80 mg, 35.3% yield). MS (ESI, m / z): 611.4 [M / 2+H]+.

[0267] Step 2:

[0268] Compound 11-2 (80 mg, 0.075 mmol) was dissolved in acetonitrile (1 mL), and a 4M dioxane hydrochloride solution (1 mL, 4 mmol) was added at 20°C. The reaction mixture was stirred at 20°C for 1 hour, concentrated, and the resulting crude product was purified by preparative HPLC to afford the formate salt of compound 11 (yellow solid, 25 mg, 35.46% yield). MS (ESI, m / z): 538.9 [M / 2+H]+. 1H NMR (400MHz, MeOD) δ8.52(s,1H),7.79–7.59(m,2H),7.42–7.15(m,3H),7.00(d,J=2.5Hz,1H),5.39(d,J=53.0Hz,1H),5.08( dd,J=12.4,5.5Hz,2H),4.58(dd,J=23.7,13.5Hz,4H),4.34(dd,J=42.3,10.8Hz,3H),4.05(s,3H),3.78(dd,J=20.4,13.2Hz ,6H),3.48(s,1H),3.23(d,J=13.7Hz,2H),3.16–2.97(m,4H),2.96–2.65(m,4H),2.58(dd,J=12.2,7.5Hz,1H),2.53–2.29(m ,5H),2.23(dd,J=17.2,5.8Hz,1H),2.15–1.95(m,9H),1.80(dd,J=29.9,16.4Hz,5H),1.72–1.59(m,2H),0.94–0.74(m,3H).

[0269] Example 12

[0270] 5-(9-(2-((2S,6R,7aS)-7a-((((S)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl))-7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-6,8-difluoroquinazolin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolizin-2-yl)ethyl)-3,9-diazaspiro[5.5]undec-3-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0271] The synthetic route is as follows:

[0272] Step 1:

[0273] Compound 12-1 (76 mg, 0.18 mmol) was dissolved in methanol (10 mL), and sodium acetate (122 mg, 1.49 mmol) was added. The reaction mixture was stirred at 20°C for 10 minutes. Compound 53-7a (150 mg, 0.19 mmol) was added, and the reaction mixture was stirred at 20°C for 30 minutes. Sodium cyanoborohydride (35 mg, 0.56 mmol) was added, and the reaction mixture was stirred at 20°C for 16 hours. The reaction mixture was diluted with dichloromethane (30 mL), washed with water (10 mL x 2) and saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on a preparative plate (dichloromethane:methanol = 0-10%) to afford compound 12-2 (white solid, 100 mg, 44.8% yield). MS (ESI, m / z): 551.9 [M / 2+H]+.

[0274] Step 2:

[0275] Compound 12-2 (90 mg, 0.075 mmol) was dissolved in acetonitrile (1 mL) and a dioxane hydrochloride solution (4 M, 1 mL, 4 mmol) was added at 20°C. The reaction mixture was stirred at 20°C for 1 hour, concentrated, and the resulting crude product was purified by preparative HPLC to afford the formate salt of compound 12 (yellow solid, 60 mg, 75.75% yield). MS (ESI, m / z): 529.9 [M / 2+H]+. 1H NMR(400MHz,MeOD)δ8.53(s,1H),7.69(dd,J=18.2,9.6Hz,3H),7.40–7.19(m,4H),6.99(d,J=2.6Hz,1H),5.44(d,1H),5.12–5.08(m,1H), 4.55(dd,J=29.5,19.3Hz,5H), 4.36(d,J=10.6Hz,1H), 4.24(d,J=10.6Hz,1H), 3.85(s,2H), 3.70(dd,J=19.4,12.9Hz,2H), 3.47(dd,J=16. 4,10.8Hz,5H),3.11(dd,J=45.1,23.3Hz,7H),2.78(ddd,J=36.3,24.9,16.3Hz,4H),2.59(s,1H),2.44(d,J=9.5Hz,3H),2.30(s,1H),2.24 –2.18(m,1H),2.16–2.10(m,1H),2.06(s,1H),1.98(s,3H),1.77(d,J=13.8Hz,5H),1.70(s,3H),1.65-1.59(m,1H),0.82(t,J=7.4Hz,3H).

[0276] Example 13

[0277] (S)-3-(5-(4-((1-((2R,6R,7aS)-7a-(7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolidin-2-yl)methyl)piperidin-4-yl)methyl)piperazin-1-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione

[0278] The synthetic route is as follows:

[0279] Step 1:

[0280] To a reaction flask, (2R,7aS)-2-fluorotetrahydro-1H-pyrrolizine-7a(5H)-methanol (30 g, 179.0 mmol, 1.0 equiv), imidazole (15.4 g, 214.8 mmol, 1.2 equiv), and 300 mL of dichloromethane were added sequentially under nitrogen at 0°C. Tert-butyldiphenylsilyl chloride (67.3 g, 232.6 mmol, 1.3 equiv) was then slowly added. The resulting mixture was stirred at 25°C under nitrogen for 2 hours. The reaction progress was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, the reaction solution was quenched with saturated sodium bicarbonate solution and extracted with dichloromethane (300 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a 0% → 10% methanol / dichloromethane mobile phase gradient elution. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 52-1 (colorless oil, 63 g, 84% yield). MS (ESI, m / z): 398.2 [M+H] + ; 1 H NMR (300MHz, CDCl3) δ7.77–7.64(m,4H),7.50–7.35(m,6H),5.35–5.09(m,1H),3.49(d,J=9.6Hz,1H),3.38(d,J=9.6 Hz,1H),3.27–2.99(m,3H),2.98–2.83(m,1H),2.26–2.14(m,1H),2.13–1.94(m,2H),1.95–1.63(m,3H),1.09(s,9H).

[0281] Step 2:

[0282] Under stirring conditions at 0 degrees Celsius, a solution of compound 52-1 (39 g, 93.2 mmol, 1.0 equivalent) in carbon tetrachloride (400 ml) was added with ruthenium trichloride hydrate (4.42 g, 18.6 mmol, 0.2 equivalent) in water (400 ml) and sodium periodate (104.9 g, 465.9 mmol, 5.0 equivalent). The mixture was stirred at 25 degrees Celsius for 1 hour, and the reaction process was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After the reaction was completed, the reaction solution was cooled to room temperature and quenched with water (500 ml). The resulting mixture was extracted with dichloromethane (500 ml × 3), the organic phases were combined, and then dried over anhydrous sodium sulfate. The desiccant was filtered to remove the desiccant, and the filtrate was evaporated under reduced pressure to remove the solvent to obtain a crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 60% petroleum ether / methyl tert-butyl ether as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to afford compound 48-6 (white solid, 19 g, 47% yield). MS (ESI, m / z): 412.2 [M+H] + ; 1 H NMR (400MHz, CDCl3) δ7.66–7.59(m,4H),7.48–7.37(m,6H),5.38–5.15(m,1H),4.21–4.08(m,1H),3.63–3.53(m,1H),3.49–3.39(m, 1H),3.17–3.00(m,1H),2.79–2.66(m,1H),2.44–2.34(m,1H),2.33–2.21(m,1H),2.21–2.10(m,1H),2.05–1.92(m,2H),1.04(s,9H).

[0283] Step 3:

[0284] To a solution of compound 48-6 (16 g, 36.930 mmol, 1.0 eq) in anhydrous tetrahydrofuran (150 ml) and hexamethylphosphoric triamide (25 ml) was added dropwise a solution of lithium diisopropylamide in tetrahydrofuran (46.5 ml, 1 mol / L, 1.5 eq) under nitrogen protection and stirring at -78 degrees Celsius. The resulting mixture was reacted for 30 minutes under nitrogen protection and stirring at -78 degrees Celsius. Paraformaldehyde (3.5 g, 73.860 mmol, 2.0 eq) was added to the reaction system under nitrogen protection and stirring at -78 degrees Celsius, and the mixture was then slowly heated to 25 degrees Celsius. The mixture was reacted for 2.5 hours under nitrogen protection and stirring at 25 degrees Celsius. The reaction process was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After the reaction, saturated aqueous ammonium chloride (500 mL) was slowly added to the reaction mixture at 0°C to quench the reaction. The mixture was extracted with ethyl acetate (500 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The desiccant was filtered to remove the desiccant, and the filtrate was evaporated to remove the solvent under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography using a mobile phase gradient of 0% → 100% petroleum ether / methyl tert-butyl ether. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 52-2 (yellow oil, 5.5 g, 32% yield). MS (ESI, m / z): 442.2 [M+H] + ; 1 H NMR (400MHz, CDCl3) δ7.64–7.61(m,4H),7.47–7.38(m,6H),5.33–5.18(m,1H),4.21–4.12(m,1H),3.82–3.78(m,1H),3.68–3.62(m,1H), 3.59(d,J=10.4Hz,1H),3.44(d,J=10.4Hz,1H),3.14–2.97(m,2H),2.33–2.16(m,2H),2.06–1.94(m,1H),1.84–1.80(m,1H),1.05(s,9H).

[0285] Step 4:

[0286] To a 250 mL three-necked flask, compound 52-2 (2.8 g, 6.02 mmol, 1.0 eq), allyl bromide (0.92 g, 7.22 mmol, 1.2 eq), and N,N-dimethylformamide (39 mL) were added sequentially under nitrogen protection and stirring at 25°C. The reaction system was then cooled to zero degrees Celsius. Sodium hydride (60%, 0.28 g, 7.22 mmol, 1.2 eq) was added portionwise to the reaction system under nitrogen protection and stirring at zero degrees Celsius. The resulting mixture was slowly heated to 25°C and then reacted under nitrogen protection and stirring at 25°C for 2 hours. The reaction process was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After the reaction was completed, the reaction solution was cooled to zero degrees Celsius and quenched by adding saturated aqueous ammonium chloride (500 mL) to the reaction solution under stirring at zero degrees Celsius. The mixture was extracted with ethyl acetate (500 ml x 3), and the combined organic phases were dried over anhydrous sodium sulfate. The desiccant was removed by filtration, and the filtrate was evaporated to remove the solvent under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography using a mobile phase gradient of 0% → 40% petroleum ether / methyl tert-butyl ether. The resulting fractions were evaporated to remove the solvent under reduced pressure to obtain compound 52-3 (yellow oil, 1.57 g, 51% yield). MS (ESI, m / z): 482.2 [M+H] + ; 1 H NMR (300MHz, CDCl3) δ7.73–7.57(m,4H),7.55–7.35(m,6H),6.02–5.74(m,1H),5.47–5.08(m,3H),4.24–4.09(m,1H),4.06 –3.95(m,2H),3.86–3.75(m,1H),3.64–3.40(m,3H),3.25–3.00(m,2H),2.43–2.20(m,2H),2.12–1.90(m,2H),1.07(s,9H).

[0287] Step 5:

[0288] To a solution of compound 52-3 (1.5 g, 3.1 mmol, 1.0 eq) in N,N-dimethylformamide (15 ml) was added lithium aluminum tetrahydride (0.18 g, 4.67 mmol, 1.5 eq) in portions under nitrogen protection and stirring at 0°C. The resulting mixture was reacted at 60°C under nitrogen protection and stirring for 2 hours. The reaction process was monitored by thin-layer chromatography. After the reaction was completed, ice water (0.2 ml) was added to the reaction solution to quench the reaction, followed by 0.2 ml of 20% NaOH solution and finally 0.6 ml of water. After stirring for 15 minutes, the insoluble matter was removed by filtration, and the filter cake was washed with a tetrahydrofuran / methanol (10 / 1) mixture (10 ml x 3). The combined filtrates were evaporated to remove the solvent under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% → 10% dichloromethane / ammonia methanol. The resulting fraction was evaporated to remove the solvent under reduced pressure to afford compound 52-4 (colorless oil, 0.53 g, 75% yield). MS (ESI, m / z): 230.2 [M+H] + .

[0289] Step 6:

[0290] At 25 degrees Celsius under argon protection and stirring conditions, to a solution of compound 13-3 (1.0 g, 3.02 mmol) (reference document WO2023097227) in anhydrous acetonitrile (20 ml) were added 52-4 (0.69 g, 3.02 mmol) and N,N-diisopropylethylamine (1.17 g, 9.06 mmol) in sequence. The resulting mixture was reacted for 60 hours under argon protection and stirring conditions at 80 degrees Celsius. The reaction process was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After the reaction was completed, water (25 ml) was added to the reaction solution to quench the reaction. The resulting mixture was extracted with ethyl acetate (25 ml x 3), and the organic phases were combined; the organic phase was washed with saturated brine (30 ml x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography using a 0% → 10% methanol / dichloromethane mobile phase gradient elution. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 13-4 (yellow solid, 1.08 g). MS (ESI, m / z): 524.3 [M+H] + .

[0291] Step 7:

[0292] To a solution of compound 13-4 (0.6 g, 1.15 mmol) in tetrahydrofuran (12 mL) was added potassium phosphate (0.73 g, 3.44 mmol), compound 13-5 (0.62 g, 1.72 mmol), Ad2n-BuP-Pd-G3 (125 mg, 0.17 mmol), and water (3 mL) in sequence under argon at 25°C. The mixture was stirred at 70°C under argon for 16 hours. The reaction progress was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, the reaction solution was cooled to room temperature and quenched by the addition of water (15 mL). The resulting mixture was extracted with ethyl acetate (15 mL x 3). The organic phases were combined and washed with saturated brine (20 mL x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 13-6 (yellow solid, 660 mg). MS (ESI, m / z): 722.5 [M+H] + .

[0293] Step 8:

[0294] To a reaction flask, 13-6 (660 mg, 0.912 mmol), compound 13-7 (355 mg, 2.08 mmol), tetrakis(triphenylphosphine)palladium (210 mg, 0.18 mmol), and dichloromethane (10 mL) were added sequentially under nitrogen and stirred at 25°C. The resulting mixture was reacted at 25°C for 48 hours, with the reaction progress monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, the excess solvent was removed by concentration under reduced pressure to yield a crude product. The crude product was purified by silica gel column chromatography using a 0% → 10% methanol / dichloromethane mobile phase gradient. The resulting fraction was then evaporated to remove the solvent under reduced pressure to yield compound 13-8 (yellow solid, 560 mg). MS (ESI, m / z): 682.4 [M+H]+.

[0295] Step 9:

[0296] To a reaction flask, a solution of compound 13-8 (360 mg, 0.53 mmol) in dichloromethane (20 mL) was added sequentially with stirring at 25°C. DMP (220 mg, 0.52 mmol) and sodium bicarbonate (99 mg, 1.18 mmol) were then added. The mixture was stirred at room temperature for 2 hours, and the reaction progress was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion, an aqueous solution (5 mL) was added to quench the reaction. The resulting mixture was extracted with dichloromethane (5 mL x 3). The organic phases were combined and washed with saturated brine (5 mL x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to yield compound 13-9 (yellow solid, 158 mg). MS (ESI, m / z): 680.4 [M+H]+.

[0297] Step 10:

[0298] To a solution of compound 13-9 (90 mg, 0.13 mmol) in methanol (5 mL) was added compound 13-10 (86 mg, 0.20 mmol) and sodium acetate (54 mg, 0.66 mmol) at 25°C under argon. The mixture was stirred at room temperature for 15 minutes. Sodium cyanoborohydride (25 mg, 0.40 mmol) was then added. The mixture was stirred at room temperature for 16 hours under argon. The reaction was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, aqueous solution (5 mL) was added to quench the reaction. The resulting mixture was extracted with ethyl acetate (5 mL x 3). The organic phases were combined and washed with saturated brine (5 mL x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 13-11 (yellow solid, 50 mg). MS (ESI, m / z): 1089.4 [M+H] + .

[0299] Step 11:

[0300] Under argon protection and stirring at 25 degrees Celsius, a trifluoroacetic acid (2 ml) solution was added to a dichloromethane (2 ml) solution of compound 13-11 (50 mg, 0.046 mmol). The mixture was stirred at room temperature under argon protection for 2 hours. The reaction process was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After the reaction, the mixture was purified by high-performance liquid chromatography (HPLC) using an XBridge Prep OBD C18 Column, 30 x 150 mm, 5 μm; mobile phase A: water (10 mmol / L formic acid), mobile phase B: acetonitrile; flow rate: 60 ml / min; elution was performed with 5% → 35% phase B within 12 minutes, and the detector UV was 220 / 254 nm. The formate salt of compound 13 (white solid, 16.1 mg) was obtained. 1H NMR(400MHz, Methanol-d4)δ9.28(s,1H),7.68(t,J=7.9Hz,2H),7.37–7.20(m,2H),7.13(s,1H),7.05(t,J=2.4Hz,1H),5.59(d ,J=51.2Hz,2H),5.34(t,J=4.9Hz,1H),5.12(dd,J=13.3,5.1Hz,1H),4.65(dd,J=12.1,7.9Hz,3H),4.50–4.33(m,3H),4.06(s, 1H),3.84(d,J=25.0Hz,2H),3.77–3.43(m,8H),3.21(d,J=9.5Hz,4H),2.91(ddd,J=18.3,13.3,5.2Hz,4H),2.82–2.31(m,8H), 2.22–1.97(m,7H),1.90–1.73(m,3H),1.67–1.56(m,2H),1.37–1.26(m,7H),0.81(q,J=7.5Hz,2H).MS(ESI,m / z):1045.2[M+H] + .

[0301] Example 14

[0302] 2-(2,6-dioxopiperidin-3-yl)-5-(4-((1-(((2R,6R,7aS)-7a-((7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-(((R)-3-hydroxy-3-methylpiperidin-1-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolazin-2-yl)methyl)piperidin-4-yl)methyl(piperazin-1-yl)-6-fluoroisoindoline-1,3-dione

[0303] The synthetic route is as follows:

[0304] Step 1:

[0305] Compound 14-1 (81 mg, 0.18 mmol) and sodium acetate (48 mg, 0.59 mmol) were added to a methanol solution of compound 13-9 (5 mL) at 25°C with stirring under argon. The mixture was stirred at room temperature for 15 minutes under argon, after which sodium cyanoborohydride (22 mg, 0.35 mmol) was added. The mixture was stirred at room temperature under argon for 16 hours. The reaction progress was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, aqueous solution (5 mL) was added to quench the reaction. The resulting mixture was extracted with ethyl acetate (5 mL x 3). The organic phases were combined and washed with saturated brine (5 mL x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% → 10% methanol / dichloromethane. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 14-2 (yellow solid, 80 mg). MS (ESI, m / z): 1121.4 [M+H] + .

[0306] Step 2:

[0307] Under argon protection and stirring at 25 degrees Celsius, a trifluoroacetic acid (2 ml) solution was added to a dichloromethane (2 ml) solution of compound 14-2 (80 mg, 0.071 mmol). The mixture was stirred at room temperature under argon protection for 2 hours. The reaction process was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After the reaction, the mixture was purified by high-performance liquid chromatography using an XBridge Prep OBD C18 Column, 30 x 150 mm, 5 μm; mobile phase A: water (10 mmol / L formic acid), mobile phase B: acetonitrile; flow rate: 60 ml / min; elution was performed with 5% → 35% phase B within 12 minutes, and the detector UV was 220 / 254 nm. The hydrochloride salt of compound 14 (yellow-green solid, 26.5 mg) was obtained. 1H NMR (400MHz, Methanol-d4) δ9.23(d,J=1.6Hz,1H),7.66(dd,J=9.1,5.8Hz,1H),7.55(d,J=11.2Hz,1H),7.44(dd,J=7.4,2.8Hz,1H ),7.31–7.21(m,2H),7.05(d,J=2.6Hz,1H),5.42–5.28(m,1H),5.10(dd,J=12.5,5.4Hz,1H),4.42–4.25(m,4H),3.70–3.53(m,2H) ,3.47(d,J=10.1Hz,5H),3.27–3.21(m,4H),3.10(d,J=46.8Hz,3H),2.89–2.69(m,7H),2.61(t,J=5.0Hz,4H),2.53–2.38(m,3H),2 .36–2.08(m,9H),2.06–1.96(m,3H),1.88–1.76(m,5H),1.41(d,J=12.3Hz,2H),0.81(q,J=7.0Hz,3H).MS(ESI,m / z):1077.2[M+H] + .

[0308] Example 15

[0309] 3-(4-(4-((1-((2R,6R,7aS)-7a-(7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolidin-2-yl)methyl)piperidin-4-yl)methyl(piperazin-1-yl)phenyl)piperidine-2,6-dione

[0310] The synthetic route is as follows:

[0311] Step 1:

[0312] To a solution of compound 13-9 (82 mg, 0.12 mmol) in methanol (5 mL) was added compound 15-1 (68 mg, 0.18 mmol) and sodium acetate (48 mg, 0.59 mmol) at 25°C under argon and stirring. The mixture was stirred at room temperature for 15 minutes under argon. Sodium cyanoborohydride (22 mg, 0.35 mmol) was then added to the mixture. The mixture was stirred at room temperature under argon for 16 hours. The reaction progress was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, aqueous solution (5 mL) was added to quench the reaction. The resulting mixture was extracted with ethyl acetate (5 mL x 3). The organic phases were combined and washed with saturated brine (5 mL x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 15-2 (yellow solid, 68 mg). MS (ESI, m / z): 1034.4 [M+H] + .

[0313] Step 2:

[0314] Under argon protection and stirring at 25 degrees Celsius, a trifluoroacetic acid (2 ml) solution was added to a dichloromethane (2 ml) solution of compound 15-2 (68 mg, 0.066 mmol). The mixture was stirred at room temperature under argon protection for 2 hours. The reaction process was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After the reaction, the mixture was purified by high-performance liquid chromatography (HPLC) using an XBridge Prep OBD C18 Column, 30 x 150 mm, 5 μm; mobile phase A: water (10 mmol / L formic acid), mobile phase B: acetonitrile; flow rate: 60 ml / min; elution was performed with 5% → 35% phase B within 12 minutes, and the detector UV was 220 / 254 nm. The formate salt of compound 15 (white solid, 24.6 mg) was obtained. 1H NMR (400MHz, Methanol-d4) δ9.25 (s, 1H), 8.27 (s, 2H), 7.67 (t, J = 7.3Hz, 1H), 7.35 –6.88(m,6H),5.43(d,J=53.6Hz,2H),4.66–4.18(m,4H),3.81–3.36(m,8H),2.98(d ,J=93.8Hz,13H),2.75–2.33(m,7H),2.32–2.13(m,4H),2.09–1.76(m,6H),1.55(d, J=43.1Hz,2H),1.38–1.20(m,5H),0.81(q,J=7.4Hz,2H).MS(ESI,m / z):990.5[M+H] + .

[0315] Example 16

[0316] Compound 16: 3-(5-(4-((4-(2-((2S,6R,7aS)-7a-(((4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolazin-2-yl)ethyl)piperazin-1-yl)methyl)piperidin-1-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione

[0317] The synthetic route is as follows:

[0318] Step 1:

[0319] Compound 10-4 (58 mg, 0.14 mmol) was dissolved in methanol (10 mL), and sodium acetate (75 mg, 0.92 mmol) was added. The reaction solution was stirred at 20°C for 10 minutes. Compound 1-7 (90 mg, 0.11 mmol) was added, and the reaction solution was stirred at 20°C for 30 minutes. Sodium cyanoborohydride (22 mg, 0.34 mmol) was added, and the reaction solution was stirred at 20°C for 16 hours. The reaction solution was diluted with dichloromethane (30 mL), washed with water (10 mL x 2) and saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on a preparative plate (dichloromethane:methanol = 0-10%) to obtain compound 16-1 (white solid, 40 mg, 29.23% yield). MS (ESI, m / z): 598.9 [M / 2+H]+.

[0320] Step 2:

[0321] Compound 16-1 (45 mg, 0.038 mmol) was dissolved in acetonitrile (1 mL), and a dioxane hydrochloride solution (4 M, 1 mL, 4 mmol) was added at 20°C. The reaction mixture was stirred at 20°C for 1 hour, filtered, and the resulting crude product was purified by preparative HPLC to afford the formate salt of compound 16 (yellow solid, 25 mg, 63.17% yield). MS (ESI, m / z): 1052.3 [M+H]+.

[0322] 1H NMR (400MHz, MeOD) δ9.07(s,1H),8.48(s,1H),7.89(dd,J=9.1,5.8Hz,1H),7.64(d,J=8.4Hz,1H),7.36(dd,J=16.3 ,5.6Hz,2H),7.23(d,J=2.4Hz,1H),7.08(d,J=8.9Hz,2H),5.40(d,J=52.6Hz,1H),5.12(dd,J=13.3,5.1Hz,1H),4.7 7–4.64(m,2H),4.38(ddd,J=23.7,19.0,10.4Hz,4H),4.04–3.69(m,6H),3.56–3.41(m,2H),3.38(d,J=2.9Hz,3H),3 .26(s,1H),3.06–2.56(m,13H),2.55–2.28(m,6H),2.18(dd,J=20.7,15.3Hz,2H),2.08–1.59(m,10H),1.30(s,2H).

[0323] Example 17

[0324] Compound 17: 1-(5-(9-(2-((2S,6R,7aS)-7a-(((4-((1R,5S)-3,8-diazabicyclo[3.2.1]oct-3-yl))-7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolizin-2-yl)ethyl)-3,9-diazaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0325] The synthetic route is as follows:

[0326] Step 1:

[0327] Compound 17-1 (55 mg, 0.14 mmol) was dissolved in methanol (10 mL), and sodium acetate (75 mg, 0.92 mmol) was added. The reaction mixture was stirred at 20°C for 10 minutes. Compound 1-7 (90 mg, 0.11 mmol) was added, and the reaction mixture was stirred at 20°C for 30 minutes. Sodium cyanoborohydride (22 mg, 0.34 mmol) was added, and the reaction mixture was stirred at 20°C for 16 hours. The reaction mixture was diluted with dichloromethane (30 mL), washed with water (10 mL x 2) and saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on a preparative plate (dichloromethane:methanol = 0-10%) to afford compound 17-2 (white solid, 50 mg, 37.32% yield). MS (ESI, m / z): 586.4 [M / 2+H]+.

[0328] Step 2:

[0329] Compound 17-2 (45 mg, 0.038 mmol) was dissolved in acetonitrile (1 mL) and a dioxane hydrochloride solution (4 M, 1 mL, 4 mmol) was added at 20°C. The reaction mixture was stirred at 20°C for 1 hour, filtered, and the resulting crude product was purified by preparative HPLC to afford the formate salt of compound 17 (yellow solid, 27 mg, 68.42% yield). MS (ESI, m / z): 1027.2 [M+H]+. 1H NMR(400MHz,MeOD)δ9.06(s,1H),8.48(s,2H),7.89(dd,J=9.2,5.7Hz,1H),7.47(dd,J=8.5,2.1Hz,1H),7.37(ddd,J=11.4,10 .5,1.6Hz,3H),7.22(d,J=7.7Hz,2H),5.38(d,J=52.4Hz,1H),4.73(s,2H),4.45–4.37(m,1H),4.31–4.24(m,1H),3.95(d,J=0. 6Hz,4H),3.89–3.79(m,2H),3.74(s,2H),3.49(dd,J=11.4,9.8Hz,3H),3.37(d,J=5.2Hz,2H),3.28(d,J=14.3Hz,2H),3.15(d ,J=1.6Hz,4H),3.08(s,2H),2.83(t,J=6.7Hz,3H),2.60–2.10(m,6H),2.08–1.92(m,4H),1.79(d,J=18.6Hz,6H),1.59(s,5H).

[0330] Example 18

[0331] Compound 18: 5-(4-((1-(2-((2S,6R,7aS)-7a-((((7S)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl))-7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-6,8-difluoroquinazolin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolazin-2-yl)ethyl)piperidin-4-yl)oxy)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0332] The synthetic route is as follows:

[0333] Step 1:

[0334] Compound 18-1 (60 mg, 0.136 mmol) was dissolved in methanol (10 mL), and sodium acetate (83 mg, 1.01 mmol) was added. The reaction solution was stirred at 20°C for 10 minutes. Compound 53-7a (90 mg, 0.127 mmol) was added, and the reaction solution was stirred at 20°C for 30 minutes. Sodium cyanoborohydride (24 mg, 0.38 mmol) was added, and the reaction solution was stirred at 20°C for 16 hours. The reaction solution was diluted with dichloromethane (30 mL), washed with water (10 mL x 2) and saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on a preparative plate (dichloromethane:methanol = 0-10%) to obtain compound 18-2 (white solid, 65 mg, 45.1% yield). MS (ESI, m / z): 616.9 [M / 2+H]+.

[0335] Step 2:

[0336] Compound 18-2 (65 mg, 0.057 mmol) was dissolved in acetonitrile (1 mL) and a dioxane hydrochloride solution (4 M, 1 mL, 4 mmol) was added at 20°C. The reaction mixture was stirred at 20°C for 1 hour, concentrated, and the resulting crude product was purified by preparative HPLC to afford the formate salt of compound 18 (yellow solid, 35 mg, 55.97% yield). MS (ESI, m / z): 544.9 [M / 2+H]+. 1H NMR (400MHz, MeOD) δ8.52 (s, 1H), 7.77–7.60 (m, 3H), 7.40–7.17 (m, 4H), 7.00 (d, J = 2.5Hz, 1H), 5.46 (s, 0.5H), 5.33 (s, 0.5H), 5.0 8(dd,J=12.4,5.5Hz,1H),4.57(d,J=10.2Hz,1H),4.39(d,J=10.8Hz,1H),4.29(d,J=10.9Hz,1H),4.05(s,1H),3.86–3.75(m,2H) ,3.48(s,1H),3.23(d,J=13.7Hz,1H),3.06(d,J=28.9Hz,1H),2.89–2.69(m,2H),2.63–2.54(m,1H),2.49–2.30(m,2H),2.23(d,J =11.3Hz,1H),2.06(dd,J=33.0,14.4Hz,3H),1.92–1.78(m,2H),1.70–1.61(m,1H),1.33(d,J=19.0Hz,1H),0.81(t,J=7.4Hz,3H)

[0337] Example 19

[0338] Compound 19: 3-(5-(4-((1-(2-((2S,6R,7aS)-7a-(((4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolazin-2-yl)ethyl)piperidin-4-yl)methyl)piperidin-1-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione

[0339] The synthetic route is as follows:

[0340] Step 1:

[0341] To a solution of compound 1-2 (1.2 g, 2.03 mmol) in tetrahydrofuran (30 mL) was added potassium phosphate (2.1 g, 9.9 mmol), compound 13-5 (1.2 g, 3.3 mmol), Ad2n-BuP-Pd-G3 (0.4 g, 0.5 mmol), and water (7 mL) in sequence under argon at 25°C. The mixture was stirred at 70°C under argon for 4 hours. The reaction process was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, the reaction solution was cooled to room temperature and quenched by the addition of water (15 mL). The resulting mixture was extracted with ethyl acetate (15 mL x 3), and the organic phases were combined; the organic phases were washed with saturated brine (20 mL x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 19-1 (yellow solid, 1.2 g). MS (ESI, m / z): 789.9 [M+H] + .

[0342] Step 2:

[0343] To a solution of compound 19-1 (1.2 g, 1.5 mmol) in dichloromethane (24 ml) was added potassium osmate dihydrate (0.12 g, 0.33 mmol), N-methylmorpholine N-oxide (0.7 g, 6.0 mmol), and water (6 ml) in sequence under argon protection and stirring at 25 degrees Celsius. The mixture was stirred at room temperature under argon protection for 16 hours. The reaction process was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After the reaction was completed, sodium sulfite (15 ml) was added to quench the reaction. The resulting mixture was extracted with dichloromethane (15 ml x 3), and the organic phases were combined; the organic phases were washed with saturated brine (20 ml x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 19-2 (yellow solid, 1.2 g). MS (ESI, m / z): 823.9 [M+H] + .

[0344] Step 3:

[0345] Under argon protection and stirring at 25 degrees Celsius, sodium periodate (560 mg, 2.62 mmol) and water (6 ml) were added to a solution of compound 19-2 (1.2 g, 1.5 mmol) in acetonitrile (24 ml). The mixture was stirred at room temperature under argon protection for 15 minutes. The reaction process was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After the reaction was completed, aqueous solution (25 ml) was added to quench the reaction. The resulting mixture was extracted with ethyl acetate (25 ml x 3), and the organic phases were combined; the organic phases were washed with saturated brine (5 ml x 1), then dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain compound 19-3 (yellow solid, 158 mg). MS (ESI, m / z): 791.7 [M+H] + .

[0346] Step 4:

[0347] To a solution of compound 19-4 (60 mg, 0.14 mmol) in methanol (5 ml) was added compound 19-3 (90 mg, 0.13 mmol) and sodium acetate (83 mg, 1.01 mmol) at 25 degrees Celsius under argon protection and stirring. The mixture was stirred at room temperature for 15 minutes under argon protection, after which sodium cyanoborohydride (24 mg, 0.38 mmol) was added to the mixture. The mixture was stirred at room temperature under argon protection for 16 hours. The reaction process was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, aqueous solution (5 ml) was added to quench the reaction. The resulting mixture was extracted with ethyl acetate (5 ml x 3), and the organic phases were combined; the organic phases were washed with saturated brine (5 ml x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 19-5 (yellow solid, 50 mg). MS (ESI, m / z): 1099.3 [M+H] + .

[0348] Step 5:

[0349] To a solution of compound 19-5 (50 mg, 0.045 mmol) in acetonitrile (2 mL) was added a solution of dioxane hydrochloride (4 mol / L, 2 mL) under argon and stirring at 25°C. The mixture was stirred at room temperature under argon for 2 hours. The reaction progress was monitored by LC / MS and thin-layer chromatography. After completion of the reaction, the mixture was purified by HPLC using an XBridge Prep OBD C18 Column, 30 x 150 mm, 5 μm; mobile phase A: water (10 mmol / L formic acid), mobile phase B: acetonitrile; flow rate: 60 mL / min; elution was performed with 5% → 35% phase B over 12 minutes; UV detector: 220 / 254 nm. This yielded the formate salt of compound 19 (pale yellow solid, 7 mg). 1H NMR(400MHz, Methanol-d4)δ9.11(s,1H),8.51(s,1H),7.79–7.60(m,1H),7.40–7.20(m,1H),7.07(d,J=2.7Hz,1H),5.44– 5.40(m,1H),5.31–5.28(m,1H),5.11(d,J=7.2Hz,1H),4.60–4.56(m,1H),4.43–4.19(m,2H),3.86(d,J=28.9Hz,2H),3.50( s,1H),3.12(d,J=23.8Hz,1H),2.87(dd,J=24.7,10.4Hz,2H),2.55–2.39(m,1H),2.19(s,1H),1.95(d,J=22.3Hz,1H),1.7 6(s,2H),1.62(s,0H),1.44(d,J=11.3Hz,1H),1.29(d,J=13.4Hz,2H),0.82(t,J=7.3Hz,3H)MS(ESI,m / z):528.3[M / 2+H]+.

[0350] Example 20

[0351] Compound 20: 5-(4-((1-(2-((2S,6R,7aS)-7a-(((4-(3,8-diazabicyclo[3.2.1]octan-3-yl))-7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolazin-2-yl)ethyl)piperidin-4-yl)oxy)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0352] The synthetic route is as follows:

[0353] Step 1:

[0354] To a solution of compound 18-1 (60 mg, 0.136 mmol) in methanol (5 ml) was added compound 19-3 (90 mg, 0.127 mmol) and sodium acetate (83 mg, 1.01 mmol) at 25°C under argon. The mixture was stirred at room temperature for 15 minutes. Sodium cyanoborohydride (24 mg, 0.382 mmol) was then added to the mixture. The mixture was stirred at room temperature for 16 hours under argon. The reaction was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, aqueous solution (5 ml) was added to quench the reaction. The resulting mixture was extracted with ethyl acetate (5 ml x 3). The organic phases were combined and washed with saturated brine (5 ml x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 20-1 (white solid, 30 mg). MS (ESI, m / z): 566.65 [M / 2+H] + .

[0355] Step 2:

[0356] To a solution of compound 20-1 (30 mg, 0.027 mmol) in acetonitrile (2 mL) was added a solution of dioxane hydrochloride (4 mol / L, 2 mL) at 25°C with stirring under argon. The mixture was stirred at room temperature for 2 hours under argon. The reaction progress was monitored by LC / MS and thin-layer chromatography. After completion of the reaction, the mixture was purified by HPLC using an XBridge Prep OBD C18 Column, 30 x 150 mm, 5 μm; mobile phase A: water (10 mmol / L formic acid), mobile phase B: acetonitrile; flow rate: 60 mL / min; elution was performed with 5% → 35% phase B over 12 minutes; UV detection: 220 / 254 nm. This yielded the formate salt of compound 20 (yellow solid, 5 mg). 1 H NMR(400MHz, Methanol-d4)δ9.12(s,1H),8.45(s,3H),7.75–7.67(m,2H),7.29(ddd,J=21.4,12.5,

[0357] 9.5Hz,4H),7.07(d,J=2.2Hz,1H),5.46(s,0.5H),5.33(s,0.5H),5.12–5.07(m,1H),4. 73(s,2H),4.36(dd,J=29.4,9.1Hz,1H),4.04(s,1H),3.83(d,J=35.6Hz,2H),3.50(s,1 H),3.12(d,J=25.1Hz,1H),2.92–2.64(m,2H),2.56–2.29(m,2H),2.21(d,J=15.1Hz,0H ),2.10–1.80(m,4H),1.68(s,1H),0.81(t,J=7.4Hz,3H).).MS(ESI,m / z):536.3[M / 2+H] +

[0358] Example 21

[0359] Compound 21: 3-(5-(1-((1-(2-((2S,6R,7aS)-7a-(((4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolazin-2-yl)ethyl)piperidin-4-yl)methyl)piperidin-4-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione

[0360] The synthetic route is as follows:

[0361] Step 1:

[0362] To a solution of compound 21-1 (60 mg, 0.14 mmol) in methanol (5 ml) was added compound 19-3 (90 mg, 0.13 mmol) and sodium acetate (83 mg, 1.102 mol) at 25 degrees Celsius under argon protection and stirring. The mixture was stirred at room temperature under argon protection for 15 minutes, after which sodium cyanoborohydride (24 mg, 0.38 mmol) was added to the mixture. The mixture was stirred at room temperature under argon protection for 16 hours. The reaction process was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, aqueous solution (5 ml) was added to quench the reaction. The resulting mixture was extracted with ethyl acetate (5 ml x 3), and the organic phases were combined; the organic phases were washed with saturated brine (5 ml x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 1-9 (yellow solid, 30 mg). MS (ESI, m / z): 1199.3 [M+H] + .

[0363] Step 2:

[0364] To a solution of compound 21-2 (30 mg, 0.027 mmol) in acetonitrile (2 mL) was added a solution of dioxane hydrochloride (4 mol / L, 2 mL) at 25°C with stirring under argon. The mixture was stirred at room temperature for 2 hours under argon. The reaction progress was monitored by LC / MS and thin-layer chromatography. After completion of the reaction, the mixture was purified by HPLC using an XBridge Prep OBD C18 Column, 30 x 150 mm, 5 μm; mobile phase A: water (10 mmol / L formic acid), mobile phase B: acetonitrile; flow rate: 60 mL / min; elution was performed with 5% → 35% phase B over 12 minutes; UV detection: 220 / 254 nm. This yielded the formate salt of compound 21 (yellow solid, 6 mg). 1H NMR(400MHz, Methanol-d4)δ9.13(s,1H),8.42(s,3H),7.82–7.67(m,3H),7.52–7.44(m,2H),7.32(dd,J=18.0,5.9Hz, 3H),7.07(s,1H),5.48(s,0.5H),5.34(s,1H),5.19–5.14(m,2H),4.50(d,J=8.2Hz,1H),4.42(s,1H),4.07(s,1H),3.9 8–3.82(m,1H),3.50(s,1H),3.03(s,1H),2.94–2.78(m,2H),2.70(d,J=15.5Hz,1H),2.62–2.47(m,2H),2.34(s,1H),2 .26–2.15(m,1H),2.02(d,J=9.9Hz,2H),1.84(s,1H),1.51(s,1H),0.81(t,J=7.3Hz,3H)).MS(ESI,m / z):1055.3[M+H] + .

[0365] Example 22

[0366] Compound 22: 3-(5-(4-((1-(2-((2S,6R,7aS)-7a-(((4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-(8-ethyl-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolazin-2-yl)ethyl)piperidin-4-yl)methyl)piperidin-1-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione

[0367] The synthetic route is as follows:

[0368] Step 1:

[0369] To a solution of compound 1-2 (800 mg, 1.53 mmol) in tetrahydrofuran (30 mL) was added potassium phosphate (1.7 g, 8.0 mmol), compound 22-1 (870 mg, 2.5 mmol), Ad2n-BuP-Pd-G3 (0.25 g, 0.34 mmol), and water (7 mL) in sequence under argon at 25°C. The mixture was stirred at 70°C under argon for 4 hours. The reaction process was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, the reaction solution was cooled to room temperature and quenched by the addition of water (15 mL). The resulting mixture was extracted with ethyl acetate (15 mL x 3), and the organic phases were combined; the organic phases were washed with saturated brine (20 mL x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 22-2 (yellow solid, 0.8 g). MS (ESI, m / z): 771.9 [M+H] + .

[0370] Step 2:

[0371] To a solution of compound 22-2 (0.8 g, 1.0 mmol) in dichloromethane (24 ml) was added potassium osmate dihydrate (0.12 g, 0.33 mmol), N-methylmorpholine N-oxide (0.7 g, 6.0 mmol), and water (6 ml) in sequence under argon at 25 degrees Celsius. The mixture was stirred at room temperature under argon for 16 hours. The reaction process was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After the reaction was completed, sodium sulfite (15 ml) was added to quench the reaction. The resulting mixture was extracted with dichloromethane (15 ml x 3), and the organic phases were combined; the organic phases were washed with saturated brine (20 ml x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 22-3 (yellow solid, 0.75 g). MS (ESI, m / z): 805.9 [M+H] + .

[0372] Step 3:

[0373] Under argon protection and stirring at 25 degrees Celsius, sodium periodate (374 mg, 1.75 mmol) and water (4 ml) were added to a solution of compound 22-3 (750 mg, 0.93 mmol) in acetonitrile (16 ml). The mixture was stirred at room temperature under argon protection for 15 minutes. The reaction process was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After the reaction was completed, aqueous solution (5 ml) was added to quench the reaction. The resulting mixture was extracted with ethyl acetate (5 ml x 3), and the organic phases were combined; the organic phases were washed with saturated brine (5 ml x 1), then dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain compound 22-4 (yellow solid, 750 mg). MS (ESI, m / z): 773.7 [M+H] + .

[0374] Step 4:

[0375] To a solution of compound 22-5 (60 mg, 0.14 mmol) in methanol (5 ml) was added compound 22-4 (90 mg, 0.116 mmol) and sodium acetate (83 mg, 1.102 mmol) at 25°C under argon protection and stirring. The mixture was stirred at room temperature for 15 minutes under argon protection. Then, sodium cyanoborohydride (24 mg, 0.382 mmol) was added to the mixture. The mixture was stirred at room temperature under argon protection for 16 hours. The reaction process was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, aqueous solution (5 ml) was added to quench the reaction. The resulting mixture was extracted with ethyl acetate (5 ml x 3), and the organic phases were combined; the organic phases were washed with saturated brine (5 ml x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 22-6 (white solid, 40 mg). MS (ESI, m / z): 1182.4 [M+H] + .

[0376] Step 5:

[0377] To a solution of compound 22-6 (40 mg, 0.034 mmol) in acetonitrile (2 mL) was added a solution of dioxane hydrochloride (4 mol / L, 2 mL) at 25°C with stirring under argon. The mixture was stirred at room temperature for 2 hours under argon. The reaction progress was monitored by LC / MS and thin-layer chromatography. After completion of the reaction, the mixture was purified by HPLC using an XBridge Prep OBD C18 Column, 30 x 150 mm, 5 μm; mobile phase A: water (10 mmol / L formic acid), mobile phase B: acetonitrile; flow rate: 60 mL / min; elution was performed with 5% → 35% phase B over 12 minutes; UV detection: 220 / 254 nm. This yielded the formate salt of compound 22 (yellow solid, 15 mg). 1 H NMR (400MHz, Methanol-d4) δ9.09 (s, 1H), 8.53 (s, 1H), 7.64 (t, J = 8.6Hz, 2H), 7.42–7.29 (m, 1H),

[0378] 7.19(d,J=7.2Hz,1H),7.06(t,J=10.4Hz,2H),5.43(s,1H),5.30(s,1H),5.13(s,1H),4.38(d,J =15.6Hz,2H),4.30–4.25(m,1H),3.90(s,1H),3.80(s,2H),3.50(s,1H),3.07(s,1H),2.86(d,J =15.1Hz,2H),2.45–2.29(m,4H),2.15(s,1H),1.94(s,2H),1.80(d,J=13.1Hz,3H),1.63(s,1H) ,1.42(d,J=10.6Hz,1H),1.34–1.24(m,3H),0.91(d,J=7.4Hz,2H).MS(ESI,m / z):519.3[M / 2+H] + .

[0379] Example 23

[0380] Compound 23: 3-(4-(1-((1-(2-((2S,6R,7aS)-7a-(((4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolazin-2-yl)ethyl)piperidin-4-yl)methyl)piperidin-4-yl)phenyl)piperidine-2,6-dione

[0381] The synthetic route is as follows:

[0382] Step 1:

[0383] To a solution of compound 23-1 (60 mg, 0.14 mmol) in methanol (5 mL) was added compound 19-3 (90 mg, 0.13 mmol) and sodium acetate (83 mg, 1.102 mmol) at 25°C under argon. The mixture was stirred at room temperature for 15 minutes. Sodium cyanoborohydride (28 mg, 0.44 mmol) was then added. The mixture was stirred at room temperature under argon for 16 hours. The reaction progress was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, aqueous solution (5 mL) was added to quench the reaction. The resulting mixture was extracted with ethyl acetate (5 mL x 3). The organic phases were combined and washed with saturated brine (5 mL x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 23-2 (white solid, 30 mg). MS (ESI, m / z): 1098.3 [M+H] + .

[0384] Step 2:

[0385] To a solution of compound 23-2 (30 mg, 0.027 mmol) in acetonitrile (2 mL) was added a solution of dioxane hydrochloride (4 mol / L, 2 mL) at 25°C with stirring under argon. The mixture was stirred at room temperature for 2 hours under argon. The reaction progress was monitored by LC / MS and thin-layer chromatography. After completion of the reaction, the mixture was purified by HPLC using an XBridge Prep OBD C18 Column, 30 x 150 mm, 5 μm; mobile phase A: water (10 mmol / L formic acid), mobile phase B: acetonitrile; flow rate: 60 mL / min; elution was performed with 5% → 35% phase B over 12 minutes; UV detection: 220 / 254 nm. This yielded the formate salt of compound 23 (yellow solid, 6 mg). 1H NMR(400MHz,Methanol-d4)δ9.09(s,1H),8.57(s,2H),7.66(s,2H),7.46–7.28(m,2H) ,7.12(d,J=54.7Hz,3H),6.89(s,1H),5.41(s,0.5H),5.28(s,0.5H),4.68(s,3H),4.2 9(d,J=32.7Hz,3H),3.76(s,7H),3.12(s,2H),2.74(s,8H),2.52–2.04(m,7H),1.85(d d,J=66.9,35.9Hz,8H),1.33(s,4H),0.92(d,J=6.4Hz,3H.MS(ESI,m / z):1055.3[M+H] + .

[0386] Example 24

[0387] Compound 24: 3-(5-(4-((4-(2-((2S,6R,7aS)-7a-(((4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolidin-2-yl)ethyl)piperazin-1-yl)methyl)piperidin-1-yl)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione

[0388] The synthetic route is as follows:

[0389] Step 1:

[0390] To a solution of compound 24-1 (5 g, 29 mmol) (the compound represented by Formula 1) in methanol (60 mL) was added dropwise thionyl chloride (22.93 g, 192.7 mmol) under an ice bath. The mixture was stirred at 80°C overnight. Thin-layer chromatography indicated the reaction was complete. The reaction solution was concentrated, and a saturated sodium bicarbonate solution was slowly added. The mixture was then extracted with ethyl acetate (50 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated to afford compound 24-2 (5.1 g, 94% yield) as a white solid. H NMR (400 MHz, CDCl3) δ 7.71 (ddd, J = 8.7, 5.2, 1.9 Hz, 1H), 7.03 (dd, J = 16.9, 8.9 Hz, 1H), 3.91 (s, 3H), 2.55 (d, J = 2.7 Hz, 3H).

[0391] Step 2:

[0392] To a mixture of compound 24-2 (5 g, 26.86 mmol) (a compound as shown in Formula 2) in 1,2-dichloroethane (80 mL) was added N-bromosuccinimide (8.1 g, 46 mmol) and benzoyl peroxide (150 mg, 0.62 mmol). The mixture was degassed and purged with nitrogen 3 times, and then the mixture was stirred at 80 ° C overnight. Thin layer chromatography indicated that the reaction was complete. The mixture was cooled to 20 ° C, then filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1: 0 to 10: 1) to obtain compound 24-3 (6.7 g, 94% yield) as a colorless oil. 1H NMR (400MHz, CDCl3) δ7.81 (ddd, J=8.8, 5.0, 2.0Hz, 1H), 7.18 (td, J=9.0, 7.8Hz, 1H), 5.01 (d, J=2.2Hz, 2H), 3.95 (s, 3H).

[0393] Step 3:

[0394] To a mixture of compound 24-3 (6.6 g, 25 mmol) (a compound as shown in formula 3) and 24-4 (4.2 g, 26 mmol) (a compound as shown in formula 4) in N, N-dimethylformamide (80 mL) was added diisopropylethylamine (13 mL, 74.8 mmol). The mixture was stirred at 40 ° C for 1 hour and then heated to 110 ° C overnight. Thin layer chromatography indicated that the reaction was complete. The mixture was poured into water (50 mL) and filtered to obtain a solid. The solid was washed with ethyl acetate, water and methanol, respectively, and vacuum dried to obtain a gray solid product compound 24-5 (5.4 g, 77% yield), LC-MS: m / z: (M + H) + =209,1H NMR (400MHz, DMSO) δ11.01(s,1H),7.69–7.56(m,2H),5.12(dd,J=13.3,5.1Hz,1H),4.63(d,J=17.6Hz,1H),4.47(d,J=1 7.5Hz,1H),2.99–2.85(m,1H),2.61(d,J=17.5Hz,1H),2.43(tt,J=13.4,6.6Hz,1H),2.02(dtd,J=12.6,5.2,2.2Hz,1H).

[0395] Step 4:

[0396] To a solution of compound 24-6 (1.5 g, 5.3 mmol) (a compound as shown in Formula 6) in dimethyl sulfoxide (15 mL) was added diisopropylethylamine (3.5 g, 20 mmol) and compound 24-5 (1 g, 3.57 mmol) (a compound as shown in Formula 5). The mixture was stirred at 120 ° C for 48 hours. LCMS showed that the reaction was complete. Water (30 mL) was added and filtered to obtain a solid. The solid was washed with water, methanol and ethyl acetate, respectively, and dried in vacuo to obtain a brown solid product compound 24-7 (0.89 g, 46% yield), LC-MS: m / z: (M + H) + =544.

[0397] Step 5:

[0398] Compound 24-7 (320 mg, 0.59 mmol) (the compound represented by Formula 7) was added to 2 ml of dichloromethane, followed by 2 ml of trifluoroacetic acid. The mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated and filtered through a reverse phase column to obtain compound 24-8 (260 mg, 81% yield) as a glassy solid. LC-MS: m / z: (M+H) + =444.3.

[0399] Step 6:

[0400] Compound 24-8 (70 mg, 0.16 mmol) (the compound represented by Formula 8) and compound 19-3 (80 mg, 0.10 mmol) (the compound represented by Formula 9) were added to 6 ml of methanol, followed by sodium acetate (33 mg, 0.4 mmol). After stirring at room temperature for 20 minutes, sodium cyanoborohydride (25 mg, 0.4 mmol) was added and stirred at room temperature overnight. The reaction solution was concentrated and filtered through a reverse phase column to obtain compound 24-9 (105 mg, 85% yield) as a white solid. LC-MS: m / z: (M+H) + =609.4.

[0401] Step 7:

[0402] Compound 24-9 (80 mg, 0.066 mmol) (the compound represented by Formula 10) was added to 3 ml of dichloromethane, followed by 5 ml of 4 M dioxane hydrochloride. The mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated and then subjected to reverse phase reaction to obtain the hydrochloride salt of compound 24 (15 mg, 20% yield) as a white solid. LC-MS: m / z: (M+H) +=1074.3, 1H NMR (400MHz, DMSO) δ10.97(s,1H),9.11(s,1H),8.22(s,1H),7.77(dd,J=9.1,6.0Hz,1H),7.47(d,J=8.1Hz,1H),7.35(dd,J=11.5,6.0Hz,2H) ,7.15(t,J=7.9Hz,1H),7.02(d,J=2.5Hz,1H),5.31(d,J=53.8Hz,1H),5.07(dd,J=13.3,5.1Hz,1H),4.48(d,J=17.0Hz,3H),4.31(d,J=16.9H z,1H),4.17(t,J=10.9Hz,1H),4.06–3.98(m,1H),3.65(t,J=11.5Hz,5H),3.47(s,2H),3.21(s,1H),3.12–3.00(m,2H),2.98–2.85(m,2H),2. 76(t,J=11.6Hz,2H),2.65–2.54(m,1H),2.46–1.94(m,20H),1.83–1.5 8(m,7H),1.52–1.40(m,3H),1.32–1.17(m,2H),0.73(t,J=7.4Hz,3H).

[0403] Example 25

[0404] Compound 25: 5-(9-(2-((2S,6R,7aS)-7a-(((4-(3,8-diazabicyclo[3.2.1]octan-3-yl))-7-(8-ethyl-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolazin-2-yl)ethyl)-3,9-diazaspiro[5.5]undec-3-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0405] The synthetic route is as follows:

[0406] Step 1:

[0407] To a solution of compound 25-1 (60 mg, 0.146 mmol) in methanol (5 mL) was added compound 22-4 (90 mg, 0.116 mmol) and sodium acetate (83 mg, 1.102 mmol) at 25°C under argon. The mixture was stirred at room temperature for 15 minutes. Sodium cyanoborohydride (24 mg, 0.382 mmol) was then added. The mixture was stirred at room temperature for 16 hours under argon. The reaction was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, aqueous solution (5 mL) was added to quench the reaction. The resulting mixture was extracted with ethyl acetate (5 mL x 3). The organic phases were combined and washed with saturated brine (5 mL x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 25-2 (white solid, 35 mg). MS (ESI, m / z): 1168.4 [M+H] + .

[0408] Step 2:

[0409] To a solution of compound 25-2 (35 mg, 0.03 mmol) in acetonitrile (2 mL) was added a solution of dioxane hydrochloride (4 mol / L, 2 mL) at 25°C with stirring under argon. The mixture was stirred at room temperature for 2 hours under argon. The reaction progress was monitored by LC / MS and thin-layer chromatography. After completion of the reaction, the mixture was purified by HPLC using an XBridge Prep OBD C18 Column, 30 x 150 mm, 5 μm; mobile phase A: water (10 mmol / L formic acid), mobile phase B: acetonitrile; flow rate: 60 mL / min; elution was performed with 5% → 35% phase B over 12 minutes; UV detection: 220 / 254 nm. This yielded the formate salt of compound 25 (yellow solid, 8 mg). 1H NMR(400MHz,Methanol-d4)δ9.09(s,1H),8.57(s,2H),7.66(s,2H),7.46–7.28(m,2H) ,7.12(d,J=54.7Hz,3H),6.89(s,1H),5.41(s,0.5H),5.28(s,0.5H),4.68(s,3H),4.2 9(d,J=32.7Hz,3H),3.76(s,7H),3.12(s,2H),2.74(s,8H),2.52–2.04(m,7H),1.85(d d,J=66.9,35.9Hz,8H),1.33(s,4H),0.92(d,J=6.4Hz,3H.MS(ESI,m / z):512.3[M / 2+H] + .

[0410] Example 26

[0411] Compound 26: 5-(4-((1-(2-((2S,6R,7aS)-7a-(((4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl))-7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolazin-2-yl)ethyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0412] The synthetic route is as follows:

[0413] Step 1:

[0414] To a solution of compound 26-1 (60 mg, 0.137 mmol) in methanol (5 mL) was added compound 19-3 (90 mg, 0.121 mmol) and sodium acetate (83 mg, 1.102 mmol) at 25°C under argon. The mixture was stirred at room temperature for 15 minutes. Sodium cyanoborohydride (24 mg, 0.382 mmol) was then added. The mixture was stirred at room temperature under argon for 16 hours. The reaction was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, aqueous solution (5 mL) was added to quench the reaction. The resulting mixture was extracted with ethyl acetate (5 mL x 3). The organic phases were combined and washed with saturated brine (5 mL x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 26-2 (white solid, 45 mg). MS (ESI, m / z): 1197.4 [M+H] + .

[0415] Step 2:

[0416] To a solution of compound 26-2 (45 mg, 0.037 mmol) in acetonitrile (2 mL) was added a solution of dioxane hydrochloride (4 mol / L, 2 mL) at 25°C with stirring under argon. The mixture was stirred at room temperature for 2 hours under argon. The reaction progress was monitored by LC / MS and thin-layer chromatography. After completion of the reaction, the mixture was purified by HPLC using an XBridge Prep OBD C18 Column, 30 x 150 mm, 5 μm; mobile phase A: water (10 mmol / L formic acid), mobile phase B: acetonitrile; flow rate: 60 mL / min; elution was performed with 5% → 35% phase B over 12 minutes; UV detection: 220 / 254 nm. This yielded the formate salt of compound 26 (yellow solid, 13 mg). 1H NMR(400MHz,Methanol-d4)δ9.09(s,1H),8.57(s,2H),7.66(s,2H),7.46–7.28(m,2H) ,7.12(d,J=54.7Hz,3H),6.89(s,1H),5.41(s,0.5H),5.28(s,0.5H),4.68(s,3H),4.29 (d,J=32.7Hz,3H),3.76(s,7H),3.12(s,2H),2.74(s,8H),2.52–2.04(m,7H),1.85(dd ,J=66.9,35.9Hz,8H),1.33(s,4H),0.92(d,J=6.4Hz,3H).MS(ESI,m / z):535.3[M / 2+H] + .

[0417] Example 27

[0418] Compound 27: 3-(6-(1-((1-(2-((2S,6R,7aS))-7a-(((4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-7-(8-ethyl-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolazin-2-yl)ethyl)piperidin-4-yl)methyl)piperidin-4-yl)-1-methyl-1H-indazol-3-yl)piperidine-2,6-dione

[0419] The synthetic route is as follows:

[0420] Step 1:

[0421] To a solution of compound 27-1 (60 mg, 0.14 mmol) in methanol (5 mL) was added compound 22-4 (90 mg, 0.116 mmol) and sodium acetate (83 mg, 1.102 mmol) at 25°C under argon. The mixture was stirred at room temperature for 15 minutes. Sodium cyanoborohydride (24 mg, 0.38 mmol) was then added. The mixture was stirred at room temperature for 16 hours under argon. The reaction progress was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, aqueous solution (5 mL) was added to quench the reaction. The resulting mixture was extracted with ethyl acetate (5 mL x 3). The organic phases were combined and washed with saturated brine (5 mL x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 27-2 (white solid, 25 mg). MS (ESI, m / z): 1181.4 [M+H] + .

[0422] Step 2:

[0423] To a solution of compound 27-2 (25 mg, 0.086 mmol) in acetonitrile (2 mL) was added a solution of dioxane hydrochloride (4 mol / L, 2 mL) at 25°C with stirring under argon. The mixture was stirred at room temperature for 2 hours under argon. The reaction progress was monitored by LC / MS and thin-layer chromatography. After completion of the reaction, the mixture was purified by HPLC using an XBridge Prep OBD C18 Column, 30 x 150 mm, 5 μm; mobile phase A: water (10 mmol / L formic acid), mobile phase B: acetonitrile; flow rate: 60 mL / min; elution was performed with 5% → 35% phase B over 12 minutes; UV detection: 220 / 254 nm. This yielded the formate salt of compound 27 (yellow solid, 7 mg). 1H NMR(400MHz,Methanol-d4)δ9.09(s,1H),8.57(s,2H),7.66(s,2H),7.46–7.28(m,2H) ,7.12(d,J=54.7Hz,3H),6.89(s,1H),5.41(s,0.5H),5.28(s,0.5H),4.68(s,3H),4.2 9(d,J=32.7Hz,3H),3.76(s,7H),3.12(s,2H),2.74(s,8H),2.52–2.04(m,7H),1.85(d d,J=66.9,35.9Hz,8H),1.33(s,4H),0.92(d,J=6.4Hz,3H.MS(ESI,m / z):518.9[M / 2+H] + .

[0424] Example 28

[0425] Compound 28: 5-(5-(2-((2S,6R,7aS)-7a-(((4-(3,8-diazabicyclo[3.2.1]octan-3-yl))-7-(8-ethyl-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)-6-fluorohexahydro-1H-pyrrolazin-2-yl)ethyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0426] The synthetic route is as follows:

[0427] Step 1:

[0428] To a solution of compound 28-1 (60 mg, 0.16 mmol) in methanol (5 mL) was added compound 22-4 (90 mg, 0.116 mmol) and sodium acetate (83 mg, 1.102 mmol) at 25°C with stirring under argon. The mixture was stirred at room temperature for 15 minutes under argon. Sodium cyanoborohydride (24 mg, 0.38 mmol) was then added. The mixture was stirred at room temperature under argon for 16 hours. The reaction progress was monitored by liquid chromatography-mass spectrometry and thin-layer chromatography. After completion of the reaction, aqueous solution (5 mL) was added to quench the reaction. The resulting mixture was extracted with ethyl acetate (5 mL x 3). The organic phases were combined and washed with saturated brine (5 mL x 1), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a gradient elution of 0% to 10% methanol / dichloromethane as the mobile phase. The resulting fraction was evaporated to remove the solvent under reduced pressure to obtain compound 28-2 (white solid, 20 mg). MS (ESI, m / z): 979.1 [M+H] + .

[0429] Step 2:

[0430] To a solution of compound 28-2 (20 mg, 0.02 mmol) in acetonitrile (2 mL) was added a solution of dioxane hydrochloride (4 mol / L, 2 mL) at 25°C under argon. The mixture was stirred at room temperature for 2 hours under argon. The reaction progress was monitored by LC / MS and thin-layer chromatography. After completion of the reaction, the mixture was purified by HPLC using an XBridge Prep OBD C18 Column, 30 x 150 mm, 5 μm; mobile phase A: water (10 mmol / L formic acid), mobile phase B: acetonitrile; flow rate: 60 mL / min; elution was performed with 5% → 35% phase B over 12 minutes; UV detection: 220 / 254 nm. This yielded the formate salt of compound 28 (yellow solid, 3 mg). 1H NMR(400MHz,Methanol-d4)δ9.09(s,1H),8.57(s,2H),7.66(s,2H),7.46–7.28(m,2H) ,7.12(d,J=54.7Hz,3H),6.89(s,1H),5.41(s,0.5H),5.28(s,0.5H),4.68(s,3H),4.2 9(d,J=32.7Hz,3H),3.76(s,7H),3.12(s,2H),2.74(s,8H),2.52–2.04(m,7H),1.85(d d,J=66.9,35.9Hz,8H),1.33(s,4H),0.92(d,J=6.4Hz,3H).MS(ESI,m / z):981.3[M+H] + .

[0431] Effect Example A

[0432] 1. Purpose of the experiment

[0433] A drug screening system based on the binding of KRAS_G12D and SOS1 was used to detect the inhibitory ability of small molecule compounds on the binding activity of KRAS-G12D and SOS1.

[0434] 2. Experimental materials and equipment

[0435] Table 1

[0436] 3. Experimental Methods

[0437] 3.1 Experimental steps:

[0438] a) BI-2852 was used as a positive control. The stock solution was diluted 3-fold to the first dilution point, and the dilutions were repeated 10 times. Similarly, the test compound was diluted 3-fold to the first dilution point, and the dilutions were repeated 11 times. Using an Echo, 0.2 μL of the serially diluted compound solution was transferred to a 384-well plate. Each compound was plated in duplicate, with a final DMSO concentration of 1%. Centrifuge at 1000 rpm / min for 1 min. Reference final concentrations were 100, 33.33, 11.11, 3.70, 1.23, 0.412, 0.137, 0.046, 0.015, 0.005, and 0 μM. Test compound final concentrations were 200, 66.67, 22.22, 7.41, 2.47, 0.27, 0.091, 0.03, 0.0152, 0.01, and 0 μM.

[0439] b) Prepare KRAS_G12D in the kit with a final concentration of 10 μM GTP in diluent, transfer 5 μL to a 384-well reaction plate, and centrifuge at 1000 rpm / min for 1 minute.

[0440] c) Transfer 5 μL of the SOS1 mixture to a 384-well plate, centrifuge at 1000 rpm for 1 min, and incubate at 25°C for 15 min.

[0441] d) Transfer 10 μL of the assay mixture to a 384-well plate, centrifuge at 1000 rpm for 1 min, and incubate at 4°C overnight.

[0442] e) Read the sample using an Envision multi-function plate reader at an excitation wavelength of 665 nm and an emission wavelength of 615 nm. The 665 / 615 ratio signal intensity is used to indicate enzyme activity.

[0443] f) Analyze the raw data.

[0444] 3.2 Experimental data processing method:

[0445] The IC50 values ​​of the compounds were fitted using the nonlinear regression equation in Graphpad Prism 8:

[0446] Negative control: DMSO

[0447] Positive control: 100 μM BI-2852

[0448] The IC of the compound was obtained using the following nonlinear fitting formula: 50 (half inhibitory concentration):

[0449] Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)*HillSlope))

[0450] X: log value of compound concentration

[0451] Y:665 / 615Ratio

[0452] Effect Example B

[0453] 1. Purpose of the experiment

[0454] A drug screening system based on the binding of KRAS_G12D and cRAF was used to detect the inhibitory ability of small molecule compounds on the binding activity of KRAS_G12D and cRAF.

[0455] 2. Experimental materials and equipment

[0456] Table 2

[0457] 3. Experimental Methods

[0458] 3.1 Experimental steps:

[0459] a) BI-2852 was used as a positive control. The stock solution was diluted 3-fold to the first dilution point, and the dilutions were repeated 10 times. Similarly, the test compound was diluted 3-fold to the first dilution point, and the dilutions were repeated 11 times. Using an Echo, 0.2 μL of the serially diluted compound solution was transferred to a 384-well plate. Each compound was replicated in duplicate, with a final DMSO concentration of 1%. Centrifuge at 1000 rpm / min for 1 min. The final concentrations of the positive control were 100, 33.33, 11.11, 3.70, 1.23, 0.412, 0.137, 0.046, 0.015, 0.005, and 0 μM. The final concentrations of the test compound were 200, 66.67, 22.22, 7.41, 2.47, 0.27, 0.091, 0.03, 0.0152, 0.01, and 0 μM.

[0460] b) Prepare KRAS_G12D in the kit with a final concentration of 10 μM GTP in diluent, transfer 5 μL to a 384-well reaction plate, and centrifuge at 1000 rpm / min for 1 minute.

[0461] c) Transfer 5 μL of the cRAF mixture to a 384-well plate, centrifuge at 1000 rpm / min for 1 min, and incubate at 25°C for 15 min.

[0462] d) Transfer 10 μL of the assay mixture to a 384-well plate, centrifuge at 1000 rpm for 1 min, and incubate at 4°C overnight.

[0463] e) Read the sample using an Envision multi-function plate reader at an excitation wavelength of 665 nm and an emission wavelength of 615 nm. The 665 / 615 ratio signal intensity is used to indicate enzyme activity.

[0464] f) Analyze the raw data.

[0465] 3.2 Experimental data processing method:

[0466] Fitting compound IC values ​​using Graphpad Prism 8 nonlinear regression equations 50 :

[0467] Negative control: DMSO

[0468] Positive control: 100 μM BI-2852

[0469] The IC50 (half maximal inhibitory concentration) of the compound was obtained using the following nonlinear fitting formula:

[0470] Y=Bottom+(Top-Bottom) / (1+10^((LogIC50-X)*HillSlope))

[0471] X: log value of compound concentration

[0472] Y:665 / 615Ratio

[0473] Effect Example C

[0474] 1 Experimental Purpose

[0475] A drug screening system based on the binding of KRAS_WT and SOS1 was used to detect the inhibitory ability of small molecule compounds on the binding activity of KRAS_WT and SOS1.

[0476] 2 Experimental materials and equipment

[0477] Table 3

[0478] 3 Experimental methods

[0479] 3.1 Experimental steps:

[0480] a) BI-2852 was used as a positive control. The stock solution was diluted 3-fold to the first dilution point, followed by dilutions 10+0. Similarly, the test compound was diluted 3-fold to the first dilution point, followed by dilutions 11+0. Using an Echo, transfer 0.2 μL of the serially diluted compound solution to a 384-well plate. Duplicate wells were prepared for each compound, with a final DMSO concentration of 1%. Centrifuge at 1000 rpm / min for 1 min. The final concentrations of the positive control were 100, 33.33, 11.11, 3.70, 1.23, 0.412, 0.137, 0.046, 0.015, 0.005, and 0 μM. The final concentrations of the test compound were 200, 66.67, 22.22, 7.41, 2.47, 0.27, 0.091, 0.03, 0.0152, 0.01, and 0 μM.

[0481] b) Prepare KRAS_WT in the kit with a final concentration of 10 μM GTP in diluent, transfer 5 μL to a 384 reaction plate, and centrifuge at 1000 rpm / min for 1 minute.

[0482] c) Transfer 5 μL of the SOS1 mixture to a 384-well plate, centrifuge at 1000 rpm for 1 min, and incubate at 25°C for 15 min.

[0483] d) Transfer 10 μL of the assay mixture to a 384-well plate, centrifuge at 1000 rpm for 1 min, and incubate at 4°C overnight.

[0484] e) Read the sample using an Envision multi-function plate reader at an excitation wavelength of 665 nm and an emission wavelength of 615 nm. The 665 / 615 ratio signal intensity is used to indicate enzyme activity.

[0485] f) Analyze the raw data.

[0486] 3.2 Experimental data processing method:

[0487] Fitting compound IC values ​​using Graphpad Prism 8 nonlinear regression equations 50 :

[0488] Negative control: DMSO

[0489] Positive control: 100 μM BI-2852

[0490] The IC of the compound was obtained using the following nonlinear fitting formula: 50 (half inhibitory concentration):

[0491] Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)*HillSlope))

[0492] X: log value of compound concentration

[0493] Y:665 / 615Ratio

[0494] Experimental results: The test results of the above effect examples AC are shown in the following table:

[0495] Table 4

[0496] Effect Example D KRAS-G12D PROTAC Molecular Degradation Assay

[0497] 1 Experimental Purpose

[0498] The degradation of KRAS protein in KRAS-G12D mutants such as PANC-1, GP2D, HPAC, ASPC-1 and HT29, MKN1 cells (KRAS-wild type) was detected by JessWB to detect the targeted degradation ability and specificity of KRAS-G12D PROTAC molecules on KRAS-G12D protein.

[0499] 2.2 Experimental materials and equipment

[0500] 2 Experimental methods

[0501] a) After 1-2 generations of adherent culture, revived PANC-1, GP2D, HPAC, ASPC-1, HT29, and MKN1 cells were seeded into 12-well plates and incubated overnight at 37°C in a 5% CO2 incubator. Compounds of the invention were added at a 30 μM concentration, diluted 1 / 3 or 1 / 4, for 7+0 doses and incubated for 48 hours.

[0502] b) Aspirate the culture medium and wash the cells once with 1 mL of 1x PBS. Add 300 μL of Tryple (Gibco) to each well and digest at 37°C for 5 minutes. Terminate the digestion by adding 900 μL of culture medium to each well. Transfer the cells to a 1.5 mL centrifuge tube and collect the cells by centrifugation at 2000 rpm for 5 minutes at 4°C. Wash the cells once with 1 mL of 1x PBS and collect the cells by centrifugation at 2000 rpm for 5 minutes at 4°C.

[0503] c) Add an appropriate amount of lysis buffer containing 1x PMSF according to the cell count, mix well, and lyse on ice for 30 minutes. Centrifuge at 13,000 rpm at 4°C for 20 minutes. Transfer the supernatant to a fresh centrifuge tube.

[0504] d) Take 20 μL of bovine serum albumin (BSA) of known different concentrations to prepare a protein quantitative standard curve; dilute the sample protein to be tested by 10 times, take 20 μL and add it to the corresponding detection well; take Pierce TM BCA Protein Assay Kit BCA quantitative reagent, 200 μL / well, avoid generating bubbles, incubate at 37°C in the dark for 30 min, and measure the optical density (OD) value at a wavelength of 562 nm.

[0505] f) An appropriate amount of 0.1x sample buffer was added to the protein sample and heated in a boiling water bath for 5 minutes to fully denature the protein. The degradation rate of KRAS-G12D protein was quantitatively analyzed using the Jess Western blotting system. The primary antibodies used were KRAS mouse antibody (Lsbio) and GAPDH mouse mAb (CST).

[0506] 3 Experimental results:

[0507] The compounds of the present invention were tested for their degradation activity in cells based on the same WB degradation experiment. 50 and Dmax, where Dmax is the maximum observed protein degradation level, DC 50is the concentration of compound required to reach 50% of Dmax; the results are shown in the table below:

[0508] Table 5 Compound Degradation Activity Data

[0509] Table 6 Compound Degradation Activity Data

Claims

1. A compound as shown in formula I, or a pharmaceutically acceptable salt thereof: in, In the compound shown in formula I: R 1 C6~C 14 The aromatic group or one or more R 1-1 Substituted C6~C 14 The aromatic group; R 1-1 are independently OH, C1-C6 alkyl, C3-C8 cycloalkyl, halogen, NH2, CN, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 alkyl substituted by one or more halogens, C1-C6 alkoxy substituted by one or more halogens, or C1-C6 alkoxy substituted by one or more R 1-1-1 Substituted C3-C8 cycloalkyl; R 1-1-1 are independently C1-C6 alkyl; R 2 is a 7-12 membered bridged heterocycloalkyl, a 5-8 membered heterocycloalkyl or is surrounded by one or more R 2-1 Substituted 5-8 membered heterocycloalkyl; the heteroatoms in the bridged heterocycloalkyl and heterocycloalkyl are independently one or more of nitrogen, oxygen or sulfur, and the number of heteroatoms is independently 1, 2, 3 or 4; R 2-1 OH or C1-C6 alkyl; X is N or CR 3 ; R 3 is a halogen; R 4 is a halogen; M is a 3- to 10-membered heterocycloalkylene group or a 3- to 10-membered heterocycloalkylene group substituted by one or more halogens; the heteroatoms in the heterocycloalkylene group are each independently one or more of nitrogen, oxygen or sulfur, and the number of heteroatoms is each independently 1, 2, 3 or 4; L is -L1-L2-L3-L4-, L1 is a C1-C6 alkylene group, L2 is a 5-6 membered heterocycloalkylene group, a 7-12 membered spiroheterocycloalkyl group or a 7-12 membered cycloheterocycloalkyl group, and L3 is a connecting bond, a C1-C6 alkylene group, an oxygen atom or L4 is a connecting bond or a 5- to 6-membered heterocycloalkylene group; the heteroatoms in the heterocycloalkylene group and the spiro heterocycloalkylene group are each independently one or more of nitrogen, oxygen or sulfur, and the number of heteroatoms is each independently 1, 2, 3 or 4; wherein L4 is connected to G; G is Wherein, the compound as shown in Formula I is not any of the following compounds:

2. The compound of formula I according to claim 1, or a pharmaceutically acceptable salt thereof: in, In the compound shown in formula I: R 1 C6~C 14 The aromatic group or one or more R 1-1 Substituted C6~C 14 The aromatic group; R 1-1 are independently OH, C1-C6 alkyl, C3-C8 cycloalkyl, halogen, NH2, CN, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 alkyl substituted by one or more halogens, C1-C6 alkoxy substituted by one or more halogens, or C1-C6 alkoxy substituted by one or more R 1-1-1 Substituted C3-C8 cycloalkyl; R 1-1-1 are independently C1-C6 alkyl; R 2 is a 7-12 membered bridged heterocycloalkyl, a 5-8 membered heterocycloalkyl, or a 2-1 Substituted 5-8 membered heterocycloalkyl; the heteroatoms in the bridged heterocycloalkyl and heterocycloalkyl are independently one or more of nitrogen, oxygen or sulfur, and the number of heteroatoms is independently 1, 2, 3 or 4; R 2-1 OH or C1-C6 alkyl; X is N or CR 3 ; R 3 is a halogen; R 4 is a halogen; M is a 3- to 10-membered heterocycloalkylene group or a 3- to 10-membered heterocycloalkylene group substituted by one or more halogens; the heteroatoms in the heterocycloalkylene group are each independently one or more of nitrogen, oxygen or sulfur, and the number of heteroatoms is each independently 1, 2, 3 or 4; L is -L1-L2-L3-L4-, L1 is a C1-C6 alkylene group, L2 is a 5-6-membered heterocycloalkylene group or a 7-12-membered spiroheterocycloalkylene group, L3 is a connecting bond, a C1-C6 alkylene group or an oxygen atom, and L4 is a connecting bond or a 5-6-membered heterocycloalkylene group; the heteroatoms in the heterocycloalkylene group and the spiroheterocycloalkylene group are each independently one or more of nitrogen, oxygen or sulfur, and the number of heteroatoms is each independently 1, 2, 3 or 4; wherein L4 is connected to G; G is Wherein, the compound as shown in Formula I is not any of the following compounds:

3. The compound of formula I according to claim 1 or claim 2, or a pharmaceutically acceptable salt thereof, characterized in that: It meets one or more of the following conditions: (1)R 1 C6~C 14 The aromatic group or one or more R 1-1 Substituted C6~C 14 The aryl group; R 1-1 are independently OH, C1-C6 alkyl, halogen or C2-C6 alkynyl; (2) In M, the one or more halogen-substituted 3- to 10-membered heterocycloalkylene radicals are Wherein the a end is connected to L; (3) L is Wherein the g end is connected to G; (4)R 2 for 4. The compound of formula I as described in claim 1 or claim 2, or a pharmaceutically acceptable salt thereof, characterized in that: It meets one or more of the following conditions: (1)R 1 In the above, C6~C 14 The aromatic group and one or more R 1-1 Substituted C6~C 14 The C6~C 14 The aryl groups are independently C6~C 10 The aryl group is phenyl or naphthyl, preferably naphthyl, for example (2)R 1-1 wherein the C1-C6 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, such as ethyl; (3)R 1-1 wherein the halogen is independently fluorine, chlorine, bromine or iodine, for example fluorine; (4)R 1-1 In the above, the C2-C6 alkynyl group is ethynyl; (5)R 2 In the 7-12 membered bridged heterocycloalkyl, the heteroatom is a nitrogen atom, the number of nitrogen atoms is 2, and one of the nitrogen atoms is connected to the quinazoline ring or the pyridopyrimidine ring; preferably, it is an 8-membered bridged heterocycloalkyl or a 9-membered bridged heterocyclyl, for example: (6)R 2 wherein the 5-8 membered heterocycloalkyl group and one or more R 2-1 In the substituted 5-8 membered heterocycloalkyl, the heteroatom is a nitrogen atom, the number of nitrogen atoms is 1, and the nitrogen atom is connected to the quinazoline ring or the pyridopyrimidine ring; preferably a 6 membered heterocycloalkyl, for example (7)R 2-1 wherein the C1-C6 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, such as methyl; (8)R 3 , R 4 wherein the halogen is independently fluorine, chlorine, bromine or iodine, for example fluorine; (9) In M, the 3- to 10-membered heterocycloalkylene group and the 3- to 10-membered heterocycloalkylene group substituted with one or more halogens are independently 5- to 8-membered monocyclic heterocycloalkylene groups or in In the above, ring A and ring B are each independently a 3-5-membered saturated heterocyclic ring, the type of heteroatom in the saturated heterocyclic ring is nitrogen, oxygen or sulfur, and the number of heteroatoms is 1 or 2; Y is C or a heteroatom; preferably, ring A and ring B are each independently a 5-membered saturated heterocyclic ring, the heteroatom in the 5-membered saturated heterocyclic ring is, for example, nitrogen, and the number of heteroatoms is, for example, 1; more preferably, ring A and ring B are each independently a tetrahydropyrrole ring; (10) In M, the halogen in the 3- to 10-membered heterocycloalkylene group substituted by one or more halogens is fluorine, chlorine, bromine or iodine, for example, fluorine; (11) In L1 and L3, the C1-C6 alkylene group is independently -CH2-, -CH2CH2-, -CH2CH2CH2-, -CH(CH3)CH2- or -CH2CH(CH3)-, for example -CH2- or -CH2CH2-; (12) In L2, the 5- to 6-membered heterocycloalkylene group is independently a 6-membered heterocycloalkylene group; (13) In L2, the heteroatom of the 5- to 6-membered heterocycloalkylene group is independently nitrogen; (14) In L2, the number of heteroatoms in the 5- to 6-membered heterocycloalkylene group is independently 1 or 2; (15) In L4, the 5- to 6-membered heterocycloalkylene group is a 6-membered heterocycloalkylene group; (16) In L4, the heteroatom of the 5- to 6-membered heterocycloalkylene group is nitrogen; (17) In L4, the number of heteroatoms of the 5- to 6-membered heterocycloalkylene group is 2; (18) In L2, the 7-12-membered spiroheterocycloalkyl is an 11-membered spiroheterocycloalkyl; (19) In L2, the heteroatom of the 7-12-membered spiroheterocycloalkyl is nitrogen; (20) In L2, the number of heteroatoms of the 7-12-membered spiroheterocycloalkyl group is 2; (21) In L2, the 7-12-membered heterocycloalkyl group is an 8-membered heterocycloalkyl group; (22) In L2, the heteroatom of the 7-12-membered heterocycloalkyl group is nitrogen; (23) In L2, the number of heteroatoms of the 7-12-membered heterocycloalkyl group is 2.

5. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 4, characterized in that: It meets one or more of the following conditions: (1) In M, the 3- to 10-membered heterocycloalkylene group and the 3- to 10-membered heterocycloalkylene group substituted with one or more halogens are independently tetrahydropyrrolylene or hexahydro-1H-pyrrolizinylene, for example Wherein the a end is connected to L; (2) In M, the one or more halogen-substituted 3- to 10-membered heterocycloalkylene radicals are Wherein the a end is connected to L; (3) In L2, the 5- to 6-membered heterocycloalkylene group is a piperidinylene group or a piperazinylene group, for example The d end is connected to L1; (4) In L2, the 7- to 12-membered spiroheterocycloalkyl groups are independently 11-membered spiroheterocyclyl groups, for example The d end is connected to L1; (5) In L4, the 5- to 6-membered heterocycloalkylene group is a piperazinylene group or a piperidinylene group, for example The f end is connected to L3; (6) In L2, the 7- to 12-membered heterocycloalkyl groups are independently 8-membered heterocyclic groups, for example The d end is connected to L1; (7) In L2, the heteroatoms in the 7-12 membered heterocycloalkyl group are each independently one or more of nitrogen, oxygen or sulfur, and the number of the heteroatoms is each independently 1, 2, 3 or 4.

6. The compound of formula I according to claim 1 or claim 2, or a pharmaceutically acceptable salt thereof, characterized in that: It meets one or more of the following conditions: (i): L1 is a C1-C6 alkylene group, L2 is a 5-6 membered heterocycloalkylene group, and L3 and L4 are a connecting bond; (ii): L1 is a C1-C6 alkylene group, L2 is a 5-6 membered heterocycloalkylene group, L3 is a C1-C6 alkylene group, and L4 is a 5-6 membered heterocycloalkylene group; (iii): L1 is a C1-C6 alkylene group, L2 is a 7-12 membered spiroheterocycloalkyl group, and L3 is L4 is the connection key; (iv): L1 is a C1-C6 alkylene group, L2 is a 5-6 membered heterocycloalkylene group, L3 is an oxygen atom, and L4 is a 5-6 membered heterocycloalkylene group; (v): L1 is a C1-C6 alkylene group, L2 is a 7-12 membered spiroheterocycloalkyl group, L3 is a connecting bond, and L4 is a connecting bond; (vi): L1 is a C1-C6 alkylene group, L2 is a 7-12 membered heterocycloalkyl group, L3 is a connecting bond, and L4 is a connecting bond.

7. The compound of formula I according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, characterized in that: It meets one or more of the following conditions: (1) R 1 for (2) X is N or CF; (3)M is (4) L is Preferably 8. The compound of formula I according to claim 1 or claim 2, or a pharmaceutically acceptable salt thereof, characterized in that: They are as follows: I-1, I-2, I-3, I-4 and I-5: Among them, R 1 ,X,R 4 and M as defined in claim 1 or 2; In formula I-1, G is In formula I-2, G is In formulas I-3, I-4 and I-5, G is 9. The compound of formula I according to claim 8, or a pharmaceutically acceptable salt thereof, characterized in that: It meets one or more of the following conditions: (1) In formula I-4, G is (2) In formula I-5, G is 10. The compound of formula I according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that: The compound as shown in formula I is any one of the following groups (1) and (2): Group (1): Group (2): Preferably, the pharmaceutically acceptable salt of the compound as shown in Formula I is preferably a formate or hydrochloride salt of the compound as shown in Formula I; Preferably, the number of pharmaceutically acceptable salts of the compound as shown in Formula I is 1, 2, 3 or 4.

11. A pharmaceutical composition comprising a substance X and one or more pharmaceutical excipients, wherein the substance X is a compound of formula I as described in any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof.

12. Use of a substance Y in preparing a drug, characterized in that: The drug is a drug for treating or preventing cancer, or a drug for treating or preventing cancer mediated by KRAS mutation; The cancer is preferably blood cancer, pancreatic cancer, MYH-related polyposis, colorectal cancer, lung cancer, non-small cell lung cancer, small cell lung cancer, colorectal cancer, rectal cancer or pancreatic cancer; The KRAS mutation is preferably a KRAS-G12D mutation; the cancer mediated by the KRAS mutation is preferably a blood cancer, pancreatic cancer, MYH-related polyposis, colorectal cancer or lung cancer; The substance Y is a compound as shown in formula I as described in any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described in claim 11.

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