Benzyl or thiophenemethylene substituted aminoquinazoline derivative and application thereof as SOS1 degradation agent

Abstract

The invention discloses an SOS1 degradation agent and application thereof. The invention provides a compound as shown in a formula (I), and / or a stereoisomer, an enantiomer, a diastereoisomer, a deuterated compound, a hydrate, a solvate, a prodrug and / or a pharmaceutically acceptable salt thereof. The compound provided by the invention can be used as an SOS1 degradation agent.

Description

Benzyl or thienylmethylidene substituted aminoquinazoline derivatives and their use as SOS1 degraders

[0001] This application claims the benefit of Chinese patent application No. 2022112963993, filed on October 21, 2022. This application incorporates the entirety of the aforementioned Chinese patent application. Technical Field

[0002] The present invention belongs to the field of medical technology. Specifically, the present invention relates to an SOS1 degrader and its use as a therapeutic agent for preventing and / or treating cancer. Background Art

[0003] SOS1 (son of sevenless homolog 1) is a widely expressed regulatory protein in cells. As a key protein in signaling pathways, SOS1 plays an important role in regulating many intracellular signal transduction pathways, such as the Ras and Rac signaling pathways. SOS1 is composed of 1333 amino acids and contains a proline-rich domain (PxxP) at its C-terminus. This domain binds to growth factor receptor-bound protein 2 (Grb2), a protein in the Ras pathway, to form a Grb2-SOS1 complex. This brings SOS1 to the vicinity of Ras proteins on the cell membrane. SOS1 catalyzes the binding of Ras to GTP, promoting Ras activation and, in turn, activating multiple downstream signaling pathways, such as the Ras-Raf-Mek-Erk and Ras-PI3K-AKT-mTOR pathways. PxxP can also bind to the SH3 (Src homology 3) domain of proteins such as E3B1 in the Rac pathway to form an EPS8-E3B1-SOS1 complex. The EPS8-E3B1-SOS1 complex connects to actin filaments through EPS8, causing GTP conversion, thereby activating Rac and subsequently activating signaling pathways such as JNK and MAPK.

[0004] Ras mutations are considered major oncogenes with a high incidence in human cancers. Studies have shown that 20-30% of tumor patients harbor Ras mutations, with KRas mutations accounting for 85%, NRas and HRas mutations accounting for 12% and 3%, respectively. However, due to the picomolar affinity of GTP for its binding site and the smooth surface of the Ras protein lacking other suitable binding pockets, directly inhibiting Ras activity is considered extremely challenging.

[0005] Abnormal expression or mutation of SOS1 is also closely associated with the development of clinical diseases. Studies have shown that SOS1 mutations are present in patients with NS and CFC. HGF1 is a rare autosomal dominant genetic disease, the cause of which is also related to mutations in the PxxP domain of SOS1. In addition, abnormal expression or mutation of SOS1 is also associated with the development of cancer.

[0006] WO2018172250A1, WO2020173935A1, WO2019201848A1, WO2020180768A1, WO2020180770A1, WO2019122129A1 and EP3558979A1 disclose several types of SOS1 inhibitors, but so far, there have been no reports on SOS1 degraders. Proteolysis Targeting Chimeria (PROTAC) is a technology different from traditional small molecule inhibitors. Traditional small molecule inhibitors usually need to act on the active site of the target protein to inhibit its activity. PROTAC is a heterogeneous bifunctional molecule, one end of which is a small molecule inhibitor that can recognize the target protein through a connecting chain, and the other end is an E3 ubiquitin ligase ligand that can recognize E3 ubiquitin ligase. This bifunctional molecule recognizes the target protein and E3 ubiquitin ligase in vivo, brings the target protein and E3 ubiquitin ligase closer to form a ternary complex, ubiquitinates the target protein, and then degrades the target protein through the ubiquitin-proteasome pathway in vivo. Compared to traditional small molecule inhibitors, PROTAC only needs to bring the target protein closer to the E3 ubiquitin ligase to degrade the substrate. This mode of action allows this technology to be applied to some undruggable targets. On the other hand, since the target protein can be released after degradation and continue to participate in the degradation process of the next protein, this catalytic degradation effect allows a smaller PROTAC drug dose to achieve efficient degradation. On the other hand, traditional small molecule inhibitors are prone to drug resistance, which is often due to point mutations, causing the small molecule inhibitors to lose their inhibitory effect on the target. PROTAC can directly degrade the target protein, which can avoid the drug resistance caused by point mutations to a certain extent. Therefore, compared with traditional small molecule inhibitors, the use of PROTAC technology for the development of new drug small molecules has high advantages and feasibility.

[0007] Summary of the Invention

[0008] To address the technical problem of the lack of SOS1 protein degraders in the prior art, the present invention provides a novel SOS1 protein degrader and its application. The SOS1 protein degrader of the present invention can effectively bind to or inhibit the SOS1 target protein, and further, can effectively degrade the SOS1 protein.

[0009] The present invention solves the above technical problems through the following methods.

[0010] The present invention provides a compound of formula I, its stereoisomers, enantiomers, diastereomers, deuterated compounds, hydrates, solvates, prodrugs or pharmaceutically acceptable salts: SLE I

[0011] Among them, S is

[0012] L is L is connected to S through A or B;

[0013] E is

[0014] In a preferred embodiment of the present invention, certain groups in the compound of Formula I, its stereoisomers, enantiomers, diastereomers, deuterated forms, hydrates, solvates, prodrugs, or pharmaceutically acceptable salts thereof have the following definitions, and the definitions of the unmentioned groups are as described in any of the above schemes (this paragraph is hereinafter referred to as "in a preferred embodiment of the present invention"):

[0015] In a preferred embodiment of the present invention, S is L is

[0016] In a preferred embodiment of the present invention, S is

[0017] L is

[0018] In a preferred embodiment of the present invention, S is

[0019] L is

[0020] In a preferred embodiment of the present invention, S is

[0021] L is

[0022] In a preferred embodiment of the present invention, S is

[0023] L is

[0024] In a preferred embodiment of the present invention, S is

[0025] L is

[0026] In a preferred embodiment of the present invention, S is L is

[0027] In a preferred embodiment of the present invention, S is

[0028] L is

[0029] In a preferred embodiment of the present invention, S is L is

[0030] In a preferred embodiment of the present invention, S is

[0031] E is

[0032] In a preferred embodiment of the present invention, S is

[0033] E is

[0034] In a preferred embodiment of the present invention, S is

[0035] E is

[0036] In a preferred embodiment of the present invention, S is

[0037] E is

[0038] In a preferred embodiment of the present invention, S is

[0039] E is

[0040] In a preferred embodiment of the present invention, S is

[0041] E is

[0042] In a preferred embodiment of the present invention, S is

[0043] E is

[0044] In a preferred embodiment of the present invention, S is

[0045] E is

[0046] In a preferred embodiment of the present invention, S is E is

[0047] In a preferred embodiment of the present invention, L is E is

[0048] In a preferred embodiment of the present invention, L is E is

[0049] In a preferred embodiment of the present invention, L is E is

[0050] In a preferred embodiment of the present invention, L is E is

[0051] In a preferred embodiment of the present invention, L is E is

[0052] In a preferred embodiment of the present invention, L is E is

[0053] In a preferred embodiment of the present invention, L is E is

[0054] In a preferred embodiment of the present invention, L is E is

[0055] In a preferred embodiment of the present invention, L is E is

[0056] In a preferred embodiment of the present invention, L is E is

[0057] In a preferred embodiment of the present invention, L is E is

[0058] In a preferred embodiment of the present invention, L is E is

[0059] In a preferred embodiment of the present invention, L is E is

[0060] In a preferred embodiment of the present invention, L is E is

[0061] In a preferred embodiment of the present invention, L is E is

[0062] In a preferred embodiment of the present invention, L is E is

[0063] In a preferred embodiment of the present invention, S is L is E is

[0064] In a preferred embodiment of the present invention, S is L is E is

[0065] In a preferred embodiment of the present invention, S is L is E is

[0066] In a preferred embodiment of the present invention, S is L is E is

[0067] In a preferred embodiment of the present invention, S is L is E is

[0068] In a preferred embodiment of the present invention, S is L is E is

[0069] In a preferred embodiment of the present invention, S is L is E is

[0070] In a preferred embodiment of the present invention, S is L is E is

[0071] In a preferred embodiment of the present invention, S is L is E is

[0072] In a preferred embodiment of the present invention, A and S in L are linked.

[0073] In a preferred embodiment of the present invention, the compound represented by formula I is any of the following compounds:

[0074] The present invention also provides a pharmaceutical composition comprising G and a pharmaceutically acceptable carrier, diluent or excipient, wherein G is the above-mentioned compound, its stereoisomer, enantiomer, diastereomer, deuterated substance, hydrate, solvate, prodrug, or pharmaceutically acceptable salt.

[0075] The present invention provides a method for degrading SOS1 protein, comprising contacting the above-mentioned compound, its stereoisomer, enantiomer, diastereomer, deuterated product, hydrate, solvate, metabolite, prodrug, or pharmaceutically acceptable salt, or the above-mentioned pharmaceutical composition with SOS1 protein.

[0076] The present invention provides the above-mentioned compound, its stereoisomers, enantiomers, diastereomers, deuterated substances, hydrates, solvates, metabolites, prodrugs, or pharmaceutically acceptable salts, or the above-mentioned pharmaceutical compositions for use as drugs for treating or preventing SOS1-mediated diseases or conditions.

[0077] The present invention provides the above-mentioned compound, its stereoisomers, enantiomers, diastereomers, deuterated forms, hydrates, solvates, metabolites, prodrugs, or pharmaceutically acceptable salts, or the above-mentioned pharmaceutical compositions for use as a drug for treating or preventing diseases or conditions (cancer) caused by the interaction between SOS1 and Ras (e.g., KRAS) or SOS1 and Rac (e.g., KRAS).

[0078] The present invention provides the above-mentioned compound, its stereoisomers, enantiomers, diastereomers, deuterated forms, hydrates, solvates, metabolites, prodrugs, or pharmaceutically acceptable salts, or the use of the above-mentioned pharmaceutical composition in the preparation of a drug for treating or preventing SOS1-mediated diseases or conditions (e.g., cancer).

[0079] The present invention provides the use of the above-mentioned compound, its stereoisomers, enantiomers, diastereomers, deuterated forms, hydrates, solvates, metabolites, prodrugs, or pharmaceutically acceptable salts, or the above-mentioned pharmaceutical compositions in the preparation of drugs for treating or preventing diseases or conditions (e.g., cancer) caused by the interaction between SOS1 and Ras (e.g., KRAS) or SOS1 and Rac (e.g., KRAS).

[0080] The present invention provides the above-mentioned compound, its stereoisomers, enantiomers, diastereomers, deuterated products, hydrates, solvates, metabolites, prodrugs, or pharmaceutically acceptable salts, or the use of the above-mentioned pharmaceutical composition in the preparation of drugs for treating or preventing cancer.

[0081] In any embodiment of the present invention, the cancer can be pancreatic cancer, lung cancer, colorectal cancer, cholangiocarcinoma, multiple myeloma, melanoma, uterine cancer, endometrial cancer, thyroid cancer, acute myeloid leukemia, bladder cancer, urothelial carcinoma, gastric cancer, cervical cancer, head and neck squamous cell carcinoma, diffuse large B-cell lymphoma, esophageal cancer, chronic lymphocytic leukemia, hepatocellular carcinoma, breast cancer, ovarian cancer, prostate cancer, glioblastoma, renal cancer or sarcoma.

[0082] The present invention provides the above-mentioned compound, its stereoisomers, enantiomers, diastereomers, deuterated forms, hydrates, solvates, metabolites, prodrugs, or pharmaceutically acceptable salts, or the above-mentioned pharmaceutical compositions for use in preparing medicaments for treating or preventing pancreatic cancer, lung cancer, colorectal cancer, cholangiocarcinoma, multiple myeloma, melanoma, uterine cancer, endometrial cancer, thyroid cancer, acute myeloid leukemia, bladder cancer, urothelial carcinoma, gastric cancer, cervical cancer, head and neck squamous cell carcinoma, diffuse large B-cell lymphoma, esophageal cancer, chronic lymphocytic leukemia, hepatocellular carcinoma, breast cancer, ovarian cancer, prostate cancer, glioblastoma, renal cancer and sarcoma.

[0083] The present invention provides the above-mentioned compound, its stereoisomers, enantiomers, diastereomers, deuterated forms, hydrates, solvates, metabolites, prodrugs, or pharmaceutically acceptable salts, or the use of the above-mentioned pharmaceutical composition in the preparation of a drug for treating or preventing neurofibromatosis type 1 (NF1), Noonan syndrome (NS), Noonan syndrome with multiple lentigines (NSML), capillary malformation-arteriovenous malformation syndrome (CM-AVM), Costello syndrome (CS), cardiofacial-cutaneous syndrome (CFC), Leggs syndrome, and hereditary gingival fibromatosis.

[0084] In any embodiment of the present invention, the SOS1-mediated disease or condition may be cancer, neurofibromatosis type 1 (NF1), Noonan syndrome (NS), Noonan syndrome with multiple lentigines (NSML), capillary malformation-arteriovenous malformation syndrome (CM-AVM), Costello syndrome (CS), cardiofacial-cutaneous syndrome (CFC), Leggs syndrome or hereditary gingival fibromatosis; wherein the cancer may be the cancer described in any embodiment of the present invention.

[0085] The present invention provides a method for treating or preventing a disease or condition mediated by SOS1, comprising administering to a patient in need thereof a therapeutically effective amount of the above-mentioned compound, its stereoisomers, enantiomers, diastereomers, deuterated substances, hydrates, solvates, metabolites, prodrugs, or pharmaceutically acceptable salts thereof, or the above-mentioned pharmaceutical composition.

[0086] The present invention provides a method for treating or preventing a disease or condition (such as cancer) regulated by the interaction between SOS1 and Ras (such as KRAS) or SOS1 and Rac (such as KRAS), comprising administering to a patient in need thereof a therapeutically effective amount of the above-mentioned compound, or its stereoisomers, enantiomers, diastereomers, deuterated substances, hydrates, solvates, metabolites, prodrugs, or pharmaceutically acceptable salts thereof, or the above-mentioned pharmaceutical composition.

[0087] In certain embodiments of the present invention, the cancer may be selected from:

[0088] Heart: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma, and teratoma group;

[0089] Lung: bronchial cancer (squamous cell carcinoma, undifferentiated small cell carcinoma, undifferentiated large cell carcinoma, adenocarcinoma), alveolar carcinoma (bronchiolar carcinoma), bronchial adenoma, sarcoma, lymphoma, enchondroma, hamartoma, mesothelioma;

[0090] Gastrointestinal tract: esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumor, vipoma), small intestine (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large intestine (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma);

[0091] Genitourinary system: kidney (adenocarcinoma, Wilms' tumor, lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonal carcinoma, teratoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenomatoid tumor, lipoma);

[0092] Liver: liver cancer (hepatocellular carcinoma), bile duct cancer, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma;

[0093] Biliary tract: gallbladder cancer, ampullary cancer, bile duct cancer;

[0094] Bone: osteosarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cell sarcoma), multiple myeloma, malignant giant cell chordoma, enchondroma (osteochondral exostosis), benign enchondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma, and giant cell tumor;

[0095] Nervous system: skull (osteomas, hemangiomas, granulomas, xanthomas, osteitis deformans), meninges (meningiomas, meningeal sarcomas, gliomas), brain (astrocytomas, medulloblastomas, gliomas, ependymomas, germinal cell tumors (pinealomas), glioblastomas multiforme, oligodendrogliomas, schwannomas, retinoblastomas, congenital tumors), spinal neurofibromas, meningiomas, gliomas, sarcomas);

[0096] Gynecology: uterus (endometrial cancer (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosa-thecal cell tumor, Sertoli-Leydig cell tumor, germ cell tumor, malignant teratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma)), fallopian tube (cancer);

[0097] Hematology: Blood (myeloid leukemia (acute and chronic), acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative disorders, multiple myeloma, myelodysplastic syndrome), Hodgkin's disease, non-Hodgkin's lymphoma (malignant lymphoma);

[0098] Skin: malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, nevus dysplasia, lipoma, hemangioma, dermatofibroma, keloid, psoriasis;

[0099] Adrenal gland: neuroblastoma.

[0100] In certain embodiments of the present invention, the cancer can be selected from pancreatic cancer, lung cancer, colorectal cancer, cholangiocarcinoma, multiple myeloma, melanoma, uterine cancer, endometrial cancer, thyroid cancer, acute myeloid leukemia, bladder cancer, urothelial carcinoma, gastric cancer, cervical cancer, head and neck squamous cell carcinoma, diffuse large B-cell lymphoma, esophageal cancer, chronic lymphocytic leukemia, hepatocellular carcinoma, breast cancer, ovarian cancer, prostate cancer, glioblastoma, renal cancer and sarcoma.

[0101] The present invention also provides a use of the above-mentioned compound, or its stereoisomers, enantiomers, diastereomers, deuterated compounds, hydrates, solvates, prodrugs, or pharmaceutically acceptable salts or pharmaceutical compositions in the preparation of a medicament for treating and / or preventing KRAS-mediated diseases or conditions.

[0102] The present invention also provides a method for treating and / or preventing KRAS-mediated diseases or conditions, comprising administering to a patient in need thereof a therapeutically effective amount of the above-mentioned compound, or its stereoisomers, enantiomers, diastereomers, deuterated substances, hydrates, solvates, metabolites, prodrugs, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof.

[0103] In certain embodiments of the present invention, the KRAS may be a mutant KRAS, preferably one or more of KRAS G12C, KRAS G12D, KRAS G13D, and KRAS G12V.

[0104] DETAILED DESCRIPTION: Unless otherwise stated, the following terms used in the specification and claims have the following meanings.

[0105] A "pharmaceutical composition" refers to a mixture containing one or more compounds described herein, or their physiologically / pharmaceutically acceptable salts or prodrugs, together with other chemical components, and other components such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, promote absorption of the active ingredient, and thereby exert its biological activity.

[0106] The present invention also provides pharmaceutically acceptable salts of compounds of formula (I). The term "pharmaceutically acceptable salt" refers to acid addition salts or base addition salts of relatively nontoxic compounds of the present invention. The acid addition salts are salts formed between compounds of formula (I) of the present invention and suitable inorganic or organic acids. These salts can be prepared during the final separation and purification of the compound, or can be prepared by reacting the purified compound of formula (I) in its free base form with a suitable organic or inorganic acid. Representative acid addition salts include hydrochloride, hydrobromide, sulfate, bisulfate, sulfite, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, hydrogen phosphate, carbonate, bicarbonate, toluate, citrate, maleate, fumarate, succinate, benzoate, tartrate, methanesulfonate, p-toluenesulfonate, gluconate, lactobionate, and laurylsulfonate. The base addition salt is a salt formed by the compound of formula (I) with a suitable inorganic base or organic base, including, for example, salts formed with alkali metals, alkaline earth metals, and quaternary ammonium cations, such as sodium salts, lithium salts, potassium salts, calcium salts, magnesium salts, tetramethylammonium salts, tetraethylammonium salts, etc.; amine salts include salts formed with ammonia (NH3), primary amines, secondary amines or tertiary amines, such as methylamine salts, dimethylamine salts, trimethylamine salts, triethylamine salts, ethylamine salts, etc.

[0107] The compounds of the present invention or their pharmaceutically acceptable salts can be administered to mammals, including humans, orally, rectally, parenterally (intravenously, intramuscularly or subcutaneously), topically (powders, ointments or drops), or intratumorally.

[0108] The compound of the present invention can be administered at a dosage of approximately 0.05-300 mg / kg body weight / day, preferably 10-300 mg / kg body weight / day, more preferably 10-200 mg / kg body weight / day.

[0109] The compounds of the present invention or pharmaceutically acceptable salts thereof can be formulated into solid dosage forms for oral administration, including but not limited to capsules, tablets, pills, powders, granules and the like. In these solid dosage forms, the compound of formula (I) of the present invention is mixed as an active ingredient with at least one conventional inert excipient (or carrier), for example, sodium citrate or dicalcium phosphate, or with the following ingredients: (1) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol and silicic acid; (2) binders, such as hydroxymethyl cellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and gum arabic; (3) humectants, such as glycerol; (4) disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates and sodium carbonate; (5) solubilizers, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, such as cetyl alcohol and glyceryl monostearate; (8) adsorbents, such as kaolin; and (9) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or a mixture thereof. Capsules, tablets and pills may also contain buffering agents.

[0110] Solid dosage forms such as tablets, sugar pills, capsules, pills, and granules can be coated or microencapsulated with coatings and shell materials such as enteric coatings and other materials known in the art. They can contain opacifying agents, and the release of the active ingredient in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active ingredient can also be microencapsulated with one or more of the above-mentioned excipients.

[0111] The compounds of the present invention or their pharmaceutically acceptable salts can be formulated into liquid dosage forms for oral administration, including but not limited to pharmaceutically acceptable emulsions, solutions, suspensions, syrups and tinctures. In addition to the compound of formula (I) or its pharmaceutically acceptable salt as the active ingredient, the liquid dosage form may contain inert diluents conventionally used in the art, such as water and other solvents, solubilizers and emulsifiers, such as ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide, and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, or mixtures thereof. In addition to these inert diluents, the liquid dosage form of the present invention may also contain conventional adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and fragrances.

[0112] The suspending agent includes ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol, microcrystalline cellulose, aluminum methylate and agar or a mixture of these substances.

[0113] The compounds of the present invention or pharmaceutically acceptable salts thereof can be formulated into dosage forms for parenteral injection, including but not limited to physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0114] The compounds of the present invention or their pharmaceutically acceptable salts can also be formulated into dosage forms for topical administration including ointments, powders, suppositories, drops, sprays and inhalants, etc. The active ingredient, the compound of formula (I) of the present invention or its pharmaceutically acceptable salt is mixed under sterile conditions with a physiologically acceptable carrier and optional preservatives, buffers, or propellants that may be required if necessary.

[0115] The present invention also provides a pharmaceutical composition comprising a compound of formula (I) of the present invention or a pharmaceutically acceptable salt thereof as an active ingredient, and a pharmaceutically acceptable carrier, excipient, or diluent. When preparing the pharmaceutical composition, the compound of formula (I) of the present invention or a pharmaceutically acceptable salt thereof is typically mixed with a pharmaceutically acceptable carrier, excipient, or diluent.

[0116] The composition of the present invention can be formulated into conventional pharmaceutical preparations according to conventional preparation methods, such as tablets, pills, capsules, powders, granules, emulsions, suspensions, dispersions, solutions, syrups, elixirs, ointments, drops, suppositories, inhalants, sprays, etc.

[0117] The compounds of the present invention or their pharmaceutically acceptable salts can be administered alone or, if necessary, in combination with other pharmaceutically acceptable therapeutic agents, such as other anti-tumor drugs. The components to be combined can be administered simultaneously or sequentially, in the form of a single formulation or in the form of different formulations. The combination can include not only a combination of a compound of the present invention and one other active agent, but also a combination of a compound of the present invention and two or more other active agents.

[0118] In the present invention, other pharmaceutically acceptable therapeutic agents that can be used together or in combination with the SOS1 degrader compound of formula (I) may be: EGFR and / or its mutant inhibitors, ErbB2 (Her2) and / or its mutant inhibitors, ALK and / or its mutant inhibitors, MEK and / or its mutant inhibitors, Kras and / or its mutant inhibitors, BCR-ABL and / or its mutant inhibitors, FGFR1 / FGFR2 / FGFR3 and / or its mutant inhibitors, ROS1 and / or its mutant inhibitors, c-MET and / or its mutant inhibitors, AXL and / or its mutant inhibitors, NTRK1 and / or its mutant inhibitors, RET and / or its mutant inhibitors, taxanes, platinum-containing compounds, antimetabolites, mitotic kinase inhibitors, immunotherapeutic agents, anti-angiogenic drugs, topoisomerase inhibitors, A-Raf / B-Raf / C-RAf and / or its mutant inhibitors, ERK and / or its mutant inhibitors, apoptosis inhibitors, mTOR inhibitors, epigenetic regulators, IGF1 / 2 and / or IGF1-R inhibitors, Ras GEF and / or its mutant inhibitors, PI3K and / or its mutant inhibitors.

[0119] In the present invention, other pharmaceutically acceptable therapeutic agents that can be used together or in combination with the SOS1 degrading agent compound of formula (I) can be: afatinib, erlotinib, gefitinib, lapatinib, cetuximab, panitumumab, osimertinib, olmutinib, EGF-816, trastuzumab, pertuzumab, crizotinib, alectinib, entrectinib, brigatinib, ib), trametinib, cobimetinib, binimetinib, selumetinib, refametinib, imatinib, dasatinib, nilotinib, nintedanib, crizotinib, lorlatinib, ceritinib, merestinib, paclitaxel, nab-paclitaxel, docetaxel el), cisplatin, carboplatin, oxaliplatin, 5-fluorouracil, capecitabine, floxuridine, cytarabine, gemcitabine, a combination of trifluridine and tipiracil (=TAS102), palbociclib, ribociclib, abemaciclib, ipilimumab, nivolumab ), pembrolizumab, atezolizumab, avelumab, durvalumab, pidilizumab, PDR-001 (=spartalizumab), bevacizumab, irinotecan, liposomal irinotecan, topotecan, ulixertinib, rapamycin, temsirolimus, everolimus,ridaforolimus, JQ-1, GSK525762, OTX 015 (=MK8628), CPI 0610, TEN-010 (=RO6870810), xentuzumab (antibody 60833 in WO2010 / 066868), or MEDI-573 (=dusigitumab).

[0120] 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.

[0121] The reagents and raw materials used in the present invention are commercially available.

[0122] The positive progress of the present invention is that: the present invention proves through SOS1 kinase activity test experiments that the compound of formula I of the present invention can effectively bind to the SOS1 target protein or produce an inhibitory effect, and further, can effectively degrade the SOS1 protein, thereby achieving the effect of preventing or treating diseases or conditions related to SOS1 or caused by the interaction between SOS1 and Ras or SOS1 and Rac. DETAILED DESCRIPTION

[0123] 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.

[0124] Intermediate 1: 1-(2-chloro-5-(4-((2-(piperidin-4-yl)ethoxy)methyl-d2)piperidine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0125] Step 1: Preparation of benzyl 4-(2-((1-(tert-butoxycarbonyl)piperidin-4-yl)methoxy-d2)ethyl)piperidine-1-carboxylate

[0126] tert-Butyl 4-(hydroxymethyl-d2)piperidine-1-carboxylate (600 mg, 2.76 mmol) was dissolved in anhydrous THF (20 mL). NaH (60%, 133 mg, 3.31 mmol) was added at 0°C and stirred for 1 hour. Benzyl 4-(2-(tosyl)ethyl)piperidine-1-carboxylate (1.73 g, 4.14 mmol) was added, and the reaction mixture was warmed to 65°C and stirred overnight. Water (30 mL) was slowly added to the reaction mixture, and the mixture was extracted with ethyl acetate (50 mL x 3). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 1:3) to obtain benzyl 4-(2-((1-(tert-butoxycarbonyl)piperidin-4-yl)methoxy-d2)ethyl)piperidine-1-carboxylate.

[0127] LC-MS:(ESI,m / z):[M+H] + =463.2.

[0128] Step 2: Preparation of benzyl 4-(2-(piperidin-4-ylmethoxy-d2)ethyl)piperidine-1-carboxylate

[0129] Benzyl 4-(2-((1-(tert-butoxycarbonyl)piperidin-4-yl)methoxy-d2)ethyl)piperidine-1-carboxylate (350 mg, 0.758 mmol) was dissolved in dichloromethane (10 mL), and then trifluoroacetic acid (2 mL) was added and stirred at room temperature for 2 hours. The reaction solution was directly concentrated under reduced pressure to obtain benzyl 4-(2-(piperidin-4-ylmethoxy-d2)ethyl)piperidine-1-carboxylate, which was used directly in the next step without purification.

[0130] LC-MS:(ESI,m / z):[M+H] + =363.1.

[0131] Step 3: Preparation of benzyl 4-(2-((1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)piperidin-4-yl)methoxy-d2)ethyl)piperidine-1-carboxylate

[0132] Benzyl 4-(2-(piperidin-4-ylmethoxy-d2)ethyl)piperidine-1-carboxylate (350 mg, 0.758 mmol) was dissolved in dimethyl sulfoxide (5 mL), and pentafluorophenyl 4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoate (330 mg, 0.758 mmol) and DIEA (985 mg, 7.58 mmol) were added. The reaction mixture was stirred at room temperature for 2 hours. The mixture was diluted with water (50 mL) and extracted with dichloromethane (30 mL x 3). The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (ethyl acetate) to obtain benzyl 4-(2-((1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)piperidin-4-yl)methoxy-d2)ethyl)piperidine-1-carboxylate.

[0133] LC-MS:(ESI,m / z):[M+H] + =613.2.

[0134] Step 4: Preparation of 1-(2-chloro-5-(4-((2-(piperidin-4-yl)ethoxy)methyl-d2)piperidine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0135] Benzyl 4-(2-((1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)piperidin-4-yl)methoxy-d2)ethyl)piperidine-1-carboxylate (300 mg, 0.489 mmol) was added to trifluoroacetic acid (5 mL). The reaction solution was heated to 90°C and stirred for 3 hours. It was directly concentrated under reduced pressure. The residue was dissolved in dichloromethane (30 mL). Then, saturated sodium bicarbonate solution was slowly added until the solution pH = 8. The organic phase was separated and the aqueous phase was washed with dichloromethane / methanol solution (10 / 1, 40 mL x 10% ethanol). 5) extraction, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 1-(2-chloro-5-(4-((2-(piperidin-4-yl)ethoxy)methyl-d2)piperidine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione, which was used directly in the next step without purification.

[0136] LC-MS:(ESI,m / z):[M+H] + =479.1.

[0137] Example 1: (1R,4R)-4-(4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)-N-(3-(3-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)-3-azaspiro[5.5]undec-9-yl)propyl)-N-methylcyclohexane-1-carboxamide

[0138] Step 1: Preparation of tert-butyl (E)-9-(3-ethoxy-3-oxoprop-1-en-1-yl)-3-azaspiro[5.5]undecane-3-carboxylate

[0139] Phosphorylacetic acid (1.27 g, 5.7 mmol) was dissolved in THF (50 mL), and NaH (342 mg, 8.55 mmol, 60%) was added at 0°C, stirred for 2 h, and then tert-butyl 9-formyl-3-azaspiro[5.5]undecane-3-carboxylate (800 mg, 2.85 mmol) was added. Stirring was continued at room temperature for 2 h. The reaction solution was diluted with water (100 mL), extracted with ethyl acetate (100 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 1:10) to give tert-butyl (E)-9-(3-ethoxy-3-oxoprop-1-en-1-yl)-3-azaspiro[5.5]undecane-3-carboxylate.

[0140] 1 H NMR(400MHz, DMSO-d6)δ6.86(dd,J=15.8,6.9Hz,1H),5.80(dd,J=15.8,1.1Hz,1H),4.15-4.02(m,2H),3.30-3.22(m,4H), 2.23-2.05(m,1H),1.72-1.60(m,2H),1.57-1.49(m,2H),1.46-1.34(m,13H),1.32(s,1H),1.28(s,1H),1.25-1.19(m,5H).

[0141] Step 2: Preparation of tert-butyl 9-(3-ethoxy-3-oxopropyl)-3-azaspiro[5.5]undecane-3-carboxylate

[0142] (E)-9-(3-Ethoxy-3-oxoprop-1-en-1-yl)-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester (740 mg, 2.1 mmol) was dissolved in EtOH (30 mL), and Pd / C (100 mg) was added under a hydrogen atmosphere. The reaction solution was stirred at room temperature for 1 h, filtered through celite, and the filtrate was directly concentrated under reduced pressure to give 9-(3-ethoxy-3-oxopropyl)-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester.

[0143] 1 H NMR(400MHz, DMSO-d6)δ4.05(dq,J=14.2,7.2Hz,2H),3.30-3.20(m,4H),2.27(t,J=7.7Hz,2H), 1.66-1.55(m,2H),1.51-1.41(m,4H),1.40-1.32(m,11H),1.25-1.12(m,6H),1.08-0.97(m,4H).

[0144] Step 3: Preparation of tert-butyl 9-(3-hydroxypropyl)-3-azaspiro[5.5]undecane-3-carboxylate

[0145] tert-Butyl 9-(3-ethoxy-3-oxopropyl)-3-azaspiro[5.5]undecane-3-carboxylate (700 mg, 1.98 mmol) was added to THF (30 mL). A THF solution of LiBH4 (174 mg, 7.92 mmol) was added dropwise at 0°C. The reaction mixture was heated to 80°C and stirred for 16 h. The reaction mixture was diluted with water (40 mL) and extracted with ethyl acetate (40 mL x 3). The organic phase was washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 4:1) to obtain tert-butyl 9-(3-hydroxypropyl)-3-azaspiro[5.5]undecane-3-carboxylate.

[0146] 1 H NMR(400MHz,DMSO-d6)δ4.32(t,J=5.1Hz,1H),3.37-3.32(m,2H),3.29-3.15(m,4 H),1.69-1.62(m,2H),1.54-1.32(m,15H),1.26-1.10(m,6H),1.09-0.98(m,3H).

[0147] Step 4: Preparation of tert-butyl 9-(3-oxopropyl)-3-azaspiro[5.5]undecane-3-carboxylate

[0148] 9-(3-Hydroxypropyl)-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester (400 mg, 1.29 mmol) and PCC (660 mg, 2.58 mmol) were added to EA (20 mL), stirred at room temperature for 2 h, filtered through celite, and the filtrate was directly concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA=5:1) to give 9-(3-oxopropyl)-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester.

[0149] 1 H NMR(400MHz,DMSO-d6)δ9.66(s,1H),3.30-3.19(m,4H),2.46-2.35(m,2H),1.66-1.57 (m,2H),1.49-1.42(m,4H),1.42-1.33(m,11H),1.22-1.17(m,3H),1.06-0.99(m,4H).

[0150] Step 5: Preparation of tert-butyl 9-(3-(benzyl(methyl)amino)propyl)-3-azaspiro[5.5]undecane-3-carboxylate

[0151] tert-Butyl 9-(3-oxopropyl)-3-azaspiro[5.5]undecane-3-carboxylate (200 mg, 0.65 mmol), N-methyl-1-phenylmethylamine (95 mg, 0.78 mmol), and STAB (413.4 mg, 1.95 mmol) were added to THF (30 mL), and the reaction solution was stirred at 60°C for 2 h. The reaction solution was diluted with water (40 mL) and extracted with ethyl acetate (40 mL x 3). The organic phase was washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 10:1) to obtain tert-butyl 9-(3-(benzyl(methyl)amino)propyl)-3-azaspiro[5.5]undecane-3-carboxylate.

[0152] LC-MS:(ESI,m / z):[M+H] + =415.3.

[0153] Step 6: Preparation of tert-butyl 9-(3-(methylamino)propyl)-3-azaspiro[5.5]undecane-3-carboxylate

[0154] 9-(3-(Benzyl(methyl)amino)propyl)-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester (180 mg, 0.43 mmol) was added to MeOH (20 mL), and Pd(OH)2 / C (60 mg) was added under a hydrogen atmosphere. The mixture was stirred at room temperature for 2 h, filtered through celite, and the filtrate was concentrated under reduced pressure to give 9-(3-(methylamino)propyl)-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester.

[0155] LC-MS:(ESI,m / z):[M+H] + =325.3.

[0156] Step 7: Preparation of tert-butyl 9-(3-((1R,4R)-4-(4-(((R)-1-(3-((tert-butoxycarbonyl)amino)-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)-N-methylcyclohexane-1-carboxamido)propyl)-3-azaspiro[5.5]undecane-3-carboxylate

[0157] (1R,4R)-4-(4-(((R)-1-(3-((tert-butoxycarbonyl)amino)-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylic acid (200 mg, 0.33 mmol), HATU (152 mg, 0.40 mmol) and DIEA (128 mg, 0.99 mmol) were added to DMF (30 mL) and stirred at room temperature for 10 min. Then, tert-butyl 9-(3-(methylamino)propyl)-3-azaspiro[5.5]undecane-3-carboxylate (130 mg, 0.40 mmol) was added and stirring was continued for 1 hour. The reaction solution was diluted with water (40 mL) and extracted with ethyl acetate (40 mL x 3). The organic phase was washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA=10:1) to give tert-butyl 9-(3-((1R,4R)-4-(4-(((R)-1-(3-((tert-butoxycarbonyl)amino)-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)-N-methylcyclohexane-1-carboxamido)propyl)-3-azaspiro[5.5]undecane-3-carboxylate.

[0158] LC-MS:(ESI,m / z):[M+H] + =909.5.

[0159] Step 8: Preparation of (1R,4R)-N-(3-(3-azaspiro[5.5]undec-9-yl)propyl)-4-(4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)-N-methylcyclohexane-1-carboxamide

[0160] tert-Butyl 9-(3-((1R,4R)-4-(4-(((R)-1-(3-((tert-butoxycarbonyl)amino)-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)-N-methylcyclohexane-1-carboxamido)propyl)-3-azaspiro[5.5]undecane-3-carboxylate (170 mg, 0.19 mmol) was added to a HCl / distilled water solution. The reaction mixture was added into a solution of oxadiazine (4M, 5 mL), stirred at room temperature for 2 h, and concentrated under reduced pressure to obtain (1R,4R)-N-(3-(3-azaspiro[5.5]undec-9-yl)propyl)-4-(4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)-N-methylcyclohexane-1-carboxamide, which was used directly in the next step.

[0161] Step 9: Preparation of (1R,4R)-4-(4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)-N-(3-(3-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)-3-azaspiro[5.5]undec-9-yl)propyl)-N-methylcyclohexane-1-carboxamide

[0162] Pentafluorophenyl 4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoate (73.6 mg, 0.17 mmol), DIEA (65.8 mg, 0.51 mmol) and (1R,4R)-N-(3-(3-azaspiro[5.5]undec-9-yl)propyl)-4-(4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)-N-methylcyclohexane-1-carboxamide (120 mg, 0.17 mmol) were added to DMSO (10 mL). The reaction solution was stirred at room temperature for 30 min, diluted with water (20 mL), and ethyl acetate (20 mL x 10 mL) was added. 3) extraction, the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by Prep-HPLC to give (1R,4R)-4-(4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)-N-(3-(3-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)-3-azaspiro[5.5]undec-9-yl)propyl)-N-methylcyclohexane-1-carboxamide.

[0163] 1 H NMR (400MHz, DMSO-d6) δ10.51(s,1H),8.12(s,1H),8.05(d,J=6.8Hz,1H),7.63(d,J=8.2Hz,1H) ,7.54(s,1H),7.38(d,J=5.5Hz,1H),6.99(s,1H),6.88(s,1H),6.85(s,1H),6.70(s,1H),5.66- 5.40(m,3H),3.89(s,3H),3.79-3.70(m,1H),3.66-3.46(m,4H),3.28-3.22(m,2H),3.10-2.86( m,4H),2.90-2.55(m,5H),2.36(s,3H),1.9-1.75(m,4H),1.72-1.39(m,14H),1.35-0.96(m,9H).

[0164] Example 2: 1-(2-chloro-5-(4-((2-(1-((1R,4R)-4-(7-methoxy-2-methyl-4-(((R)-1-(4-(2-((methylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)quinazolin-6-yl)cyclohexane-1-carbonyl)piperidin-4-yl)ethoxy)methyl)piperidin-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0165] Step 1: Preparation of tert-butyl 4-(2-(pyridin-4-ylmethoxy)ethyl)piperidine-1-carboxylate

[0166] tert-Butyl 4-(2-hydroxyethyl)piperidine-1-carboxylate (2.00 g, 8.72 mmol) was dissolved in DMF (25 mL) and cooled to 0-5°C. NaH (1.74 g, 43.6 mmol, 60%) was added portionwise, followed by stirring at room temperature for 30 minutes. The mixture was stirred at 0-5°C for 30 minutes, followed by the addition of 4-(bromomethyl)pyridine hydrobromide (2.64 g, 10.46 mmol) and stirring at room temperature for 3 hours. The mixture was diluted with aqueous NH4Cl (100 mL) and extracted with ethyl acetate (100 mL x 3). The combined organic phases were washed with brine (100 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EtOAc = 50:1 to 1:1) to afford tert-butyl 4-(2-(pyridin-4-ylmethoxy)ethyl)piperidine-1-carboxylate.

[0167] Step 2: Preparation of tert-butyl 4-(2-(piperidin-4-ylmethoxy)ethyl)piperidine-1-carboxylate

[0168] To a solution of tert-butyl 4-(2-(pyridin-4-ylmethoxy)ethyl)piperidine-1-carboxylate (1.2 g, 3.74 mmol) in i-PrOH / H₂O (20 mL / 20 mL) was added Pd / C (150 mg), and the mixture was reacted at 65°C overnight under a hydrogen balloon. The reaction solution was filtered through Celite, and the filtrate was concentrated to afford tert-butyl 4-(2-(piperidin-4-ylmethoxy)ethyl)piperidine-1-carboxylate.

[0169] LC-MS:(ESI,m / z):[M+H] + =327.2.

[0170] Step 3: Preparation of tert-butyl 4-(2-((1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)piperidin-4-yl)methoxy)ethyl)piperidine-1-carboxylate

[0171] To a solution of tert-butyl 4-(2-(piperidin-4-ylmethoxy)ethyl)piperidine-1-carboxylate (200 mg, 0.61 mmol) in DMSO (2 mL) was added pentafluorophenyl 4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoate (280 mg, 1.64 mmol) and DIEA (237 mg, 1.83 mmol). The reaction mixture was stirred at room temperature for 2 hours. Water (20 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (20 mL x 2). The organic phases were combined, washed with saturated brine (30 mL x 3), and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated and purified by silica gel column chromatography (DCM:MeOH=100:1-10:1) to give tert-butyl 4-(2-((1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)piperidin-4-yl)methoxy)ethyl)piperidine-1-carboxylate.

[0172] LC-MS:(ESI,m / z):[M+Na] + =599.2.

[0173] Step 4: Preparation of 1-(2-chloro-5-(4-((2-(piperidin-4-yl)ethoxy)methyl)piperidine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0174] Tert-butyl 4-(2-((1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)piperidin-4-yl)methoxy)ethyl)piperidine-1-carboxylate (250 mg, crude) was dissolved in DCM (4.5 mL) and TFA (1.5 mL) was added dropwise. The reaction solution was stirred at room temperature for 2 hours and concentrated to afford 1-(2-chloro-5-(4-((2-(piperidin-4-yl)ethoxy)methyl)piperidine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione, which was used directly in the next reaction.

[0175] LC-MS:(ESI,m / z):[M+H] + =477.1.

[0176] Step 5: Preparation of tert-butyl (2-(5-((R)-1-((6-((1R,4R)-4-(4-(2-((1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)piperidin-4-yl)methoxy)ethyl)piperidine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)thiophen-3-yl)benzyl)(methyl)carboxylate

[0177] 1-(2-chloro-5-(4-((2-(piperidin-4-yl)ethoxy)methyl)piperidine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione (150 mg, 0.27 mmol) was dissolved in DMSO (4 mL), and pentafluorophenyl (1R,4R)-4-(4-(((R)-1-(4-(2-(((tert-butoxycarbonyl)(methyl)amino))methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylate (232 mg, 0.28 mmol) and DIEA (105 mg, 0.81 mmol) were added. The reaction solution was stirred at room temperature for 2 hours. Water (20 mL) was added to the reaction solution, followed by extraction with ethyl acetate (20 mL x 2). The combined organic phases were washed with saturated brine (30 mL x 3) and dried over anhydrous sodium sulfate. The mixture was filtered, the filtrate was concentrated, and the crude product was purified by silica gel column chromatography (DCM:MeOH = 100:1 to 5:1) to obtain tert-butyl (2-(5-((R)-1-((6-((1R,4R)-4-(4-(2-((1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)piperidin-4-yl)methoxy)ethyl)piperidine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)thiophen-3-yl)benzyl)(methyl)carboxylate.

[0178] LC-MS: (ESI, m / z): [(M-Boc) / 2+H] + =502.2.

[0179] Step 6: Preparation of 1-(2-chloro-5-(4-((2-(1-((1R,4R)-4-(7-methoxy-2-methyl-4-(((R)-1-(4-(2-((methylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)quinazolin-6-yl)cyclohexane-1-carbonyl)piperidin-4-yl)ethoxy)methyl)piperidine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0180] To a solution of tert-butyl (2-(5-((R)-1-((6-((1R,4R)-4-(4-(2-((1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)piperidin-4-yl)methoxy)ethyl)piperidine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)thiophen-3-yl)benzyl)(methyl)carboxylate (130 mg, crude) in DCM (3 mL) was added TFA (1 mL). The reaction solution was stirred at room temperature for 2 hours, and the reaction solution was directly concentrated under reduced pressure. The crude product was purified by pre-HPLC to give 1-(2-chloro-5-(4-((2-(1-((1R,4R)-4-(7-methoxy-2-methyl-4-(((R)-1-(4-(2-((methylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)quinazolin-6-yl)cyclohexane-1-carbonyl)piperidin-4-yl)ethoxy)methyl)piperidine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione.

[0181] LC-MS:(ESI,m / z):[M / 2+H] + =502.2.

[0182] 1 H NMR(400MHz,CD3OD)δ7.99(s,1H),7.67–7.59(m,1H),7.55–7.26(m,6H),7 .22–7.11(m,2H),7.01(s,1H),6.06(q,J=6.5Hz,1H),4.66–4.49(m,2H),4. 08–3.92(m,4H),3.79–3.68(m,4H),3.49(t,J=6.1Hz,2H),3.18–3.01(m,3 H),2.91–2.48(m,8H),2.21(s,3H),2.01–1.49(m,20H),1.38–1.11(m,6H).

[0183] Example 3: 2-(2,6-dioxopiperidin-3-yl)-5-(4-((2-(1-((1R,4R)-4-(7-methoxy-2-methyl-4-(((R)-1-(4-(2-((methylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)quinazolin-6-yl)cyclohexane-1-carbonyl)piperidin-4-yl)ethoxy)methyl)piperidin-1-yl)isoindoline-1,3-dione

[0184] Step 1: Preparation of tert-butyl 4-(2-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methoxy)ethyl)piperidine-1-carboxylate

[0185] Dissolve tert-butyl 4-(2-(piperidin-4-ylmethoxy)ethyl)piperidine-1-carboxylate (200 mg, 0.61 mmol) in DMSO (4 mL), add 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (186 mg, 0.67 mmol) and DIEA (237 mg, 1.83 mmol). The mixture is reacted at 70°C for 16 hours. After cooling to room temperature, water (20 mL) is added to the reaction solution, and the mixture is extracted with ethyl acetate (20 mL x 2). The organic phases are combined, washed with saturated sodium chloride aqueous solution (30 mL x 3), and dried over anhydrous sodium sulfate. The reaction mixture was filtered and the filtrate was concentrated. The crude product was purified by silica gel column chromatography (DCM:MeOH=100:1-20:1) to give tert-butyl 4-(2-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methoxy)ethyl)piperidine-1-carboxylate.

[0186] LC-MS:(ESI,m / z):[M-Boc+H] + =483.1.

[0187] Step 2: Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-(4-((2-(piperidin-4-yl)ethoxy)methyl)piperidin-1-yl)isoindoline-1,3-dione

[0188] Tert-butyl 4-(2-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methoxy)ethyl)piperidine-1-carboxylate (240 mg, crude) was dissolved in DCM (4.5 mL) and TFA (1.5 mL) was added. The resulting reaction solution was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was concentrated to obtain 2-(2,6-dioxopiperidin-3-yl)-5-(4-((2-(piperidin-4-yl)ethoxy)methyl)piperidin-1-yl)isoindolin-1,3-dione.

[0189] LC-MS:(ESI,m / z):[M+H] + =483.2.

[0190] Step 3: Preparation of tert-butyl (2-(5-((1R)-1-((6-((1R,4R)-4-(4-(2-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methoxy)ethyl)piperidine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)thiophen-3-yl)benzyl)(methyl)carboxylate

[0191] To a solution of 2-(2,6-dioxopiperidin-3-yl)-5-(4-((2-(piperidin-4-yl)ethoxy)methyl)piperidin-1-yl)isoindoline-1,3-dione (150 mg, 0.31 mmol) in DMSO (4 mL) was added pentafluorophenyl (1R,4R)-4-(4-(((R)-1-(4-(2-(((tert-butoxycarbonyl)(methyl)amino))methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylate (285 mg, 0.35 mmol) and DIEA (136 mg, 1.05 mmol). The mixture was stirred at room temperature for 2 hours. Water (20 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (20 mL x 2). The organic phases were combined, washed with saturated brine (30 mL x 3), and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated. The crude product was purified by silica gel column chromatography (DCM:MeOH = 100:1 to 5:1) to obtain tert-butyl (2-(5-((1R)-1-((6-((1R,4R)-4-(4-(2-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methoxy)ethyl)piperidine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)thiophen-3-yl)benzyl)(methyl)carboxylate.

[0192] LC-MS:(ESI,m / z):[M / 2+H] + =555.3.

[0193] Step 4: Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-(4-((2-(1-((1R,4R)-4-(7-methoxy-2-methyl-4-(((R)-1-(4-(2-((methylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)quinazolin-6-yl)cyclohexane-1-carbonyl)piperidin-4-yl)ethoxy)methyl)piperidin-1-yl)isoindoline-1,3-dione

[0194] Tert-butyl (2-(5-((1R)-1-((6-((1R,4R)-4-(4-(2-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methoxy)ethyl)piperidine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)thiophen-3-yl)benzyl)(methyl)carboxylate (180 mg, crude) was dissolved in DCM (1 mL) and TFA (0.5 mL) was added. The mixture was stirred at room temperature for 2 hours, concentrated, and the crude product was purified by pre-HPLC to give 2-(2,6-dioxopiperidin-3-yl)-5-(4-((2-(1-((1R,4R)-4-(7-methoxy-2-methyl-4-(((R)-1-(4-(2-((methylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)quinazolin-6-yl)cyclohexane-1-carbonyl)piperidin-4-yl)ethoxy)methyl)piperidin-1-yl)isoindoline-1,3-dione.

[0195] LC-MS:(ESI,m / z):[M+H] + =1009.3.

[0196] 1 H NMR (400MHz, CD3OD) δ8.12(s,1H),7.67–7.60(m,1H),7.57–7.52(m,1H),7.46–7.38(m,3H),7.32(t,J =2.2Hz,1H),7.27(s,1H),7.22–7.16(m,2H),7.03(s,1H),6.11(q,J=6.8Hz,1H),4.64–4.46(m,4H),4 .21(s,2H),4.05(d,J=13.2Hz,2H),3.97(s,3H),3.59–3.54(m,2H),3.23–2.93(m,6H),2.83–2.65(m, 4H),2.59–2.50(m,6H),1.94–1.80(m,10H),1.73–1.66(m,4H),1.57–1.54(m,2H),1.41–1.33(m,8H).

[0197] Example 4: 1-(2-chloro-5-(4-(4-(4-((1R,4R)-4-(7-methoxy-2-methyl-4-(((R)-1-(4-(2-((methylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)quinazolin-6-yl)cyclohexane-1-carbonyl)piperazin-1-yl)n-butyl)piperidine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0198] Step 1: Preparation of 4-(pyridin-4-yl)butanal

[0199] 4-(Pyridin-4-yl)butan-1-ol (2.50 g, 16.55 mmol) was dissolved in DCM (50 mL) and cooled to 0-10°C. Dess-Martin periodinane (8.42 g, 19.86 mmol) was added portionwise. The reaction was allowed to react at room temperature for 3 hours. The mixture was quenched by adding aqueous sodium thiosulfate and aqueous sodium bicarbonate, and extracted with DCM (30 mL x 2). The organic phase was washed with saturated NaCl solution, dried over anhydrous NaSO, filtered, and the filtrate was concentrated. The crude product was purified by silica gel column chromatography (DCM:MeOH = 50:1) to provide 4-(pyridin-4-yl)butanal.

[0200] 1 H NMR (400MHz, CDCl3) δ9.79 (s, 1H), 8.52 (d, J = 5.6Hz, 2H), 7.13 (d, J = 5.6Hz, 2H), 2.66 (t, J = 7.2Hz, 2H), 2.50 (t, J = 7.2Hz, 2H), 2.03–1.94 (m, 2H).

[0201] Step 2: Preparation of tert-butyl 4-(4-(pyridin-4-yl)n-butyl)piperazine-1-carboxylate

[0202] To a mixture of 4-(pyridin-4-yl)butanal (1.60 g, 10.73 mmol) and tert-butyl piperazine-1-carboxylate (3.00 g, 16.10 mmol) in 1,2-dichloroethane / methanol (28 mL / 14 mL) was added acetic acid (3 mL). The mixture was stirred at 30°C for 1 hour. NaBH(OAc)3 (4.55 g, 21.48 mmol) was added at 0°C, and the mixture was allowed to react at room temperature for 16 hours. After completion of the reaction, H2O (10 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (30 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The crude product was purified by silica gel column chromatography (DCM:MeOH = 20:1) to yield tert-butyl 4-(4-(pyridin-4-yl)n-butyl)piperazine-1-carboxylate.

[0203] LC-MS:(ESI,m / z):[M+H] + =320.2.

[0204] Step 3: Preparation of tert-butyl 4-(4-(piperidin-4-yl)n-butyl)piperazine-1-carboxylate

[0205] To a solution of tert-butyl 4-(4-(pyridin-4-yl)n-butyl)piperazine-1-carboxylate (1.10 g, 3.44 mmol) in i-PrOH / H₂O (20 mL / 20 mL) was added Pd / C (150 mg), and the mixture was reacted at 75°C under a hydrogen balloon for 16 hours. The reaction solution was filtered through celite, and the filtrate was concentrated to afford tert-butyl 4-(4-(piperidin-4-yl)n-butyl)piperazine-1-carboxylate, which was used directly in the next reaction.

[0206] LC-MS:(ESI,m / z):[M+H] + =326.3.

[0207] Step 4: Preparation of tert-butyl 4-(4-(1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)piperidin-4-yl)n-butyl)piperazine-1-carboxylate

[0208] To a solution of tert-butyl 4-(4-(piperidin-4-yl)-butyl)piperazine-1-carboxylate (300 mg, 0.92 mmol) in DMSO (6 mL) were added pentafluorophenyl 4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoate (401 mg, 0.92 mmol) and DIEA (238 mg, 1.84 mmol), and the reaction solution was stirred at room temperature for 2 hours. Water (20 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL x 2). The organic phases were combined and washed with saturated brine (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (DCM:MeOH=100:1 to 10:1) to give tert-butyl 4-(4-(1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)piperidin-4-yl)n-butyl)piperazine-1-carboxylate.

[0209] LC-MS:(ESI,m / z):[M+H] + =576.0.

[0210] Step 5: Preparation of 1-(2-chloro-5-(4-(4-(piperazin-1-yl)n-butyl)piperidine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0211] Tert-butyl 4-(4-(1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)piperidin-4-yl)n-butyl)piperazine-1-carboxylate (150 mg, 0.315 mmol) was dissolved in DCM (3 mL), and TFA (1.5 mL) was added dropwise. The reaction solution was stirred at room temperature for 1 hour and concentrated to afford 1-(2-chloro-5-(4-(4-(piperazin-1-yl)n-butyl)piperidine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione, which was used directly in the next reaction.

[0212] LC-MS:(ESI,m / z):[M+H] + =476.0.

[0213] Step 6: Preparation of tert-butyl (2-(5-((R)-1-((6-((1R,4R)-4-(4-(4-(1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)piperidin-4-yl)n-butyl)piperazine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)thiophen-3-yl)benzyl)(methyl)carboxylate

[0214] 1-(2-chloro-5-(4-(4-(piperazin-1-yl)n-butyl)piperidine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione (280 mg, crude) was dissolved in DMSO (4 mL), and (1R,4R)-4-(4-((R)-1-(4-(2-((tert-butoxycarbonyl)(methyl)amino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylate (255 mg, 0.315 mmol) and DIEA (203 mg, 1.575 mmol) were added. The reaction mixture was stirred at room temperature for 2 hours. Water (20 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (20 mL x 2). The organic phases were combined, washed with saturated brine (30 mL x 3), and dried over anhydrous sodium sulfate. The reaction mixture was filtered and the filtrate was concentrated. The crude product was purified by silica gel column chromatography (DCM:MeOH=100:1~5:1) to give tert-butyl (2-(5-((R)-1-((6-((1R,4R)-4-(4-(4-(1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)piperidin-4-yl)n-butyl)piperazine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)thiophen-3-yl)benzyl)(methyl)carboxylate.

[0215] LC-MS:(ESI,m / z):[M+H] +=1102.5.

[0216] Step 7: Preparation of 1-(2-chloro-5-(4-(4-(4-((1R,4R)-4-(7-methoxy-2-methyl-4-(((R)-1-(4-(2-((methylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)quinazolin-6-yl)cyclohexane-1-carbonyl)piperazin-1-yl)n-butyl)piperidine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0217] To a solution of tert-butyl (2-(5-((R)-1-((6-((1R,4R)-4-(4-(4-(1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)piperidin-4-yl)n-butyl)piperazine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)thiophen-3-yl)benzyl)(methyl)carboxylate (140 mg, 0.13 mmol) in dichloromethane (2.0 mL) was added trifluoroacetic acid (1.0 mL), and the mixture was stirred at 30°C for 1 hour. After completion of the reaction, the mixture was concentrated under reduced pressure. The crude product was purified by pre-HPLC to give 1-(2-chloro-5-(4-(4-(4-((1R,4R)-4-(7-methoxy-2-methyl-4-(((R)-1-(4-(2-((methylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)quinazolin-6-yl)cyclohexane-1-carbonyl)piperazin-1-yl)n-butyl)piperidine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione.

[0218] LC-MS:(ESI,m / z):[M+H] + =1002.3.

[0219] 1 H NMR (400MHz, CDCl3) δ7.54–7.49(m,2H),7.41–7.29(m,6H),7.22–7.12(m,3H),6.50-6.23(m,1H),6.09–6.03(m,1H),4.75–4.52( s,1H),3.91(s,3H),3.85(s,2H),3.78–3.45(m,8H),3.08–2.31(m,24H),2.00–1.83(m,4H),1.59–1.42(m,5H),1.37–0.89(m,8H).

[0220] Example 5: 3-(4-(9-((4-((1R,4R)-4-(7-methoxy-2-methyl-4-(((R)-1-(4-(2-((methylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)quinazolin-6-yl)cyclohexane-1-carbonyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)phenyl)piperidine-2,6-dione

[0221] Step 1: Preparation of 4-(2,6-di(benzyloxy)pyridin-3-yl)benzoic acid

[0222] To a mixed solution of 2,6-bis(benzyloxy)-3-bromopyridine (2.80 g, 7.57 mmol) and 4-(4,4,5,5-tetramethyl-1,3,2-dioxybenzofuran-2-yl)benzoic acid (1.87 g, 11.35 mmol) in dioxane / H₂O (50 mL / 10 mL) were added Pd(dppf)Cl₂ (262 mg, 0.36 mmol) and sodium carbonate (2.01 g, 18.92 mmol). The mixture was reacted at 80°C under nitrogen for 16 hours, cooled to room temperature, adjusted to pH 2 with 1N aqueous HCl, and extracted with ethyl acetate (30 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (DCM:MeOH=100:1-2:1) to give 4-(2,6-di(benzyloxy)pyridin-3-yl)benzoic acid.

[0223] LC-MS:(ESI,m / z):[M+H] + =412.1.

[0224] Step 2: Preparation of 4-(2,6-dioxopiperidin-3-yl)benzoic acid

[0225] To a solution of 4-(2,6-di(benzyloxy)pyridin-3-yl)benzoic acid (0.50 g, 1.22 mmol) in i-PrOH (20 mL) was added Pd / C (0.2 mg), and the mixture was reacted at 40°C under a hydrogen balloon for 16 hours. The reaction solution was filtered through Celite, and the filtrate was concentrated to obtain 4-(2,6-dioxopiperidin-3-yl)benzoic acid, which was used directly in the next reaction.

[0226] LC-MS:(ESI,m / z):[MH] - =231.9.

[0227] Step 3: Preparation of benzyl 4-((3-(4-(2,6-dioxopiperidin-3-yl)benzoyl)-3-azaspiro[5.5]undec-9-yl)methyl)piperazine-1-carboxylate

[0228] 4-(2,6-Dioxopiperidin-3-yl)benzoic acid (200 mg, 0.86 mmol) and HATU (400 mg, 1.03 mmol) were dissolved in DMF (6 mL) and stirred for 3 minutes. DIEA (220 mg, 0.72 mmol) and benzyl 4-((3-azaspiro[5.5]undec-9-yl)methyl)piperazine-1-carboxylate (320 mg, 0.86 mmol) were added sequentially. The reaction mixture was stirred at 25°C for 1 hour. The reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (10 mL x 2). The organic phase was washed with brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was purified by silica gel column chromatography (DCM:MeOH=30:1) to give benzyl 4-((3-(4-(2,6-dioxopiperidin-3-yl)benzoyl)-3-azaspiro[5.5]undec-9-yl)methyl)piperazine-1-carboxylate.

[0229] LC-MS:(ESI,m / z):[M+H] + =601.1.

[0230] Step 4: Preparation of 3-(4-(9-(piperazin-1-ylmethyl)-3-azaspiro[5.5]undecane-3-carbonyl)phenyl)piperidine-2,6-dione

[0231] To a solution of benzyl 4-((3-(4-(2,6-dioxopiperidin-3-yl)benzoyl)-3-azaspiro[5.5]undec-9-yl)methyl)piperazine-1-carboxylate (230 mg, 0.38 mmol) in i-PrOH (5 mL) was added Pd / C (0.2 mg), and the mixture was reacted under a hydrogen balloon at room temperature for 2 hours. The reaction solution was filtered through celite, and the filtrate was concentrated to afford 3-(4-(9-(piperazin-1-ylmethyl)-3-azaspiro[5.5]undec-3-carbonyl)phenyl)piperidine-2,6-dione, which was used directly in the next reaction.

[0232] LC-MS:(ESI,m / z):[M+H] + =467.0.

[0233] Step 5: Preparation of tert-butyl (2-(5-((1R)-1-((6-((1R,4R)-4-(4-((3-(4-(2,6-dioxopiperidin-3-yl)benzoyl)-3-azaspiro[5.5]undec-9-yl)methyl)piperazine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)thiophen-3-yl)benzyl)(methyl)carboxylate

[0234] To a solution of 3-(4-(9-(piperazin-1-ylmethyl)-3-azaspiro[5.5]undecane-3-carbonyl)phenyl)piperidine-2,6-dione (250 mg, crude) in DMSO (2 mL) was added pentafluorophenyl (1R,4R)-4-(4-((R)-1-(4-(2-((tert-butoxycarbonyl)(methyl)amino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylate (308 mg, 0.38 mmol) and DIEA (245 mg, 1.90 mmol). The reaction mixture was stirred at room temperature for 2 hours. Water (20 mL) was added to the reaction mixture, followed by extraction with ethyl acetate (20 mL x 2). The combined organic phases were washed with saturated brine (30 mL x 3) and dried over anhydrous sodium sulfate. The reaction mixture was filtered, the filtrate was concentrated, and the crude product was purified by pre-HPLC to give tert-butyl (2-(5-((1R)-1-((6-((1R,4R)-4-(4-((3-(4-(2,6-dioxopiperidin-3-yl)benzoyl)-3-azaspiro[5.5]undec-9-yl)methyl)piperazine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)thiophen-3-yl)benzyl)(methyl)carboxylate.

[0235] LC-MS:(ESI,m / z):[M / 2+H] + =547.4.

[0236] Step 6: Preparation of 3-(4-(9-((4-((1R,4R)-4-(7-methoxy-2-methyl-4-(((R)-1-(4-(2-((methylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)quinazolin-6-yl)cyclohexane-1-carbonyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)phenyl)piperidine-2,6-dione

[0237] To a solution of tert-butyl (2-(5-((1R)-1-((6-((1R,4R)-4-(4-((3-(4-(2,6-dioxopiperidin-3-yl)benzoyl)-3-azaspiro[5.5]undec-9-yl)methyl)piperazine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)thiophen-3-yl)benzyl)(methyl)carboxylate (160 mg, 0.15 mmol) in dichloromethane (2.0 mL) was added trifluoroacetic acid (1.0 mL), and the mixture was stirred at 30° C. for 1 hour. After completion of the reaction, the mixture was concentrated under reduced pressure. The crude product was purified by pre-HPLC to give 3-(4-(9-((4-((1R,4R)-4-(7-methoxy-2-methyl-4-(((R)-1-(4-(2-((methylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)quinazolin-6-yl)cyclohexane-1-carbonyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)phenyl)piperidine-2,6-dione.

[0238] LC-MS:(ESI,m / z):[M / 2+H] + =497.4.

[0239] 1 H NMR(400MHz,CD3OD)δ7.42–7.39(m,3H),7.32–7.27(m,6H),7.24–7.12(m,2H),7 .11(s,1H),6.06–5.98(m,1H),5.66(d,J=8.0Hz,1H),3.92(s,3H),3.83–3.79(m, 1H),3.75–3.68(m,3H),3.65–3.59(m,2H),3.56–3.48(m,2H),3.42–3.26(m,2H) ,3.14–3.02(m,1H),2.78–2.12(m,17H),2.05–1.70(m,14H),1.54–1.10(m,13H).

[0240] Example 6: 1-(2-chloro-5-(4-(2-((1-((1R,4R)-4-(4-(((R)-1-(4-(2-chloro-6-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperidin-4-yl)methoxy)ethyl)piperazine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0241] Step 1: Preparation of tert-butyl 4-(2-((1-((1R,4R)-4-(4-(((R)-1-(4-(2-chloro-6-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperidin-4-yl)methoxy)ethyl)piperazine-1-carboxylate

[0242] (1R,4R)-4-(4-(((R)-1-(4-(2-chloro-6-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylic acid (145 mg, 0.24 mmol) was dissolved in DMF (5 mL), and tert-butyl 4-(2-(piperidin-4-ylmethoxy)ethyl)piperazine-1-carboxylate (80 mg, 0.24 mmol), HATU (102 mg, 0.31 mmol) and DIEA (93 mg, 0.72 mmol) were added, and the reaction solution was stirred at room temperature overnight. The reaction solution was diluted with water (50 mL) and extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (methanol:dichloromethane = 1:9) to give tert-butyl 4-(2-((1-((1R,4R)-4-(4-(((R)-1-(4-(2-chloro-6-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperidin-4-yl)methoxy)ethyl)piperazine-1-carboxylate.

[0243] LC-MS:(ESI,m / z):[M+H] + =902.5.

[0244] Step 2: Preparation of ((1R,4R)-4-(4-(((R)-1-(4-(2-chloro-6-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexyl)(4-((2-(piperazin-1-yl)ethoxy)methyl)piperidin-1-yl)methanone

[0245] Tert-butyl 4-(2-((1-((1R,4R)-4-(4-(((R)-1-(4-(2-chloro-6-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperidin-4-yl)methoxy)ethyl)piperazine-1-carboxylate (120 mg, 0.13 mmol) was dissolved in dichloromethane (5 mL) and then trifluoroacetic acid was added. Fluoroacetic acid (1 mL) was added and stirred at room temperature for 2 hours. The reaction solution was directly concentrated under reduced pressure to give ((1R,4R)-4-(4-(((R)-1-(4-(2-chloro-6-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexyl)(4-((2-(piperazin-1-yl)ethoxy)methyl)piperidin-1-yl)methanone, which was used directly in the next step without purification.

[0246] LC-MS:(ESI,m / z):[M+H] + =802.4.

[0247] Step 3: Preparation of 1-(2-chloro-5-(4-(2-((1-((1R,4R)-4-(4-(((R)-1-(4-(2-chloro-6-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperidin-4-yl)methoxy)ethyl)piperazine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0248] ((1R,4R)-4-(4-(((R)-1-(4-(2-chloro-6-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexyl)(4-((2-(piperazin-1-yl)ethoxy)methyl)piperidin-1-yl)methanone (120 mg, 0.13 mmol) was dissolved in dimethyl sulfoxide (5 mL), and pentafluorophenyl 4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoate (57 mg, 0.13 mmol) and DIEA (168 mg, 1.3 mmol) were added, and the reaction solution was stirred at room temperature for 2 hours. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by pre-HPLC to give 1-(2-chloro-5-(4-(2-((1-((1R,4R)-4-(4-(((R)-1-(4-(2-chloro-6-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperidin-4-yl)methoxy)ethyl)piperazine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione.

[0249] LC-MS:(ESI,m / z):[M+H] + =1052.4.

[0250] 1 H NMR (400MHz, DMSO-d6) δ10.51(s,1H),8.31(d,J=8.3Hz,1H),8.05(s,1H),7.63(d,J=8.2Hz,1H),7.55( s,1H),7.45-7.31(m,4H),7.24(s,1H),7.02(s,1H),6.96(s,1H),6.04-5.88(m,1H),4.44-4.34(m,1H) ,3.97-3.86(m,4H),3.78-3.71(m,1H),3.66-3.54(m,3H),3.50-3.45(m,2H),3.27-3.20(m,2H),3.15- 2.87(m,5H),2.81-2.56(m,6H),2.45-2.37(m,5H),2.01(s,6H),1.93-1.42(m,17H),1.13-0.91(m,2H).

[0251] Example 7: 1-(5-(4-(3-(4-((1R,4R)-4-(4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperazin-1-yl)propyl)piperidin-1-yl)-2-chlorophenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0252] Step 1: Preparation of 3-(1-(4-chloro-3-nitrophenyl)piperidin-4-yl)propan-1-ol

[0253] 1-Chloro-4-fluoro-2-nitrobenzene (3.0 g, 17.04 mmol) was dissolved in DMSO (30 mL), followed by the addition of 3-(piperidin-4-yl)propan-1-ol (3.66 g, 25.57 mmol) and DIEA (6.59 g, 51.12 mmol). The mixture was heated to 50°C and stirred overnight. Water (200 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (100 mL x 3). The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:4) to obtain 3-(1-(4-chloro-3-nitrophenyl)piperidin-4-yl)propan-1-ol.

[0254] LC-MS:(ESI,m / z):[M+H] + =299.1.

[0255] Step 2: Preparation of 3-(1-(4-chloro-3-nitrophenyl)piperidin-4-yl)propyl benzoate

[0256] 3-(1-(4-chloro-3-nitrophenyl)piperidin-4-yl)propan-1-ol (200 mg, 0.67 mmol) was dissolved in tetrahydrofuran (10 mL), followed by the addition of benzoyl chloride (122 mg, 0.8 mmol) and triethylamine (203 mg, 0.8 mmol). The reaction mixture was heated to 70°C and stirred overnight. Water (50 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (50 mL x 3). The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:9) to obtain 3-(1-(4-chloro-3-nitrophenyl)piperidin-4-yl)propyl benzoate.

[0257] LC-MS:(ESI,m / z):[M+H] + =403.2.

[0258] Step 3: Preparation of 3-(1-(3-amino-4-chlorophenyl)piperidin-4-yl)propyl benzoate

[0259] Dissolve 3-(1-(4-chloro-3-nitrophenyl)piperidin-4-yl)propyl benzoate (1.0 g, 2.49 mmol) in EtOH (10 mL). Add hydrazine hydrate (2 mL) and Raney Ni (solid-liquid turbid solution, 2 mL). Stir at room temperature for 1 hour, filter, and concentrate the filtrate under reduced pressure to obtain 3-(1-(3-amino-4-chlorophenyl)piperidin-4-yl)propyl benzoate. Use this product directly in the next step without purification.

[0260] LC-MS:(ESI,m / z):[M+H] + =373.1.

[0261] Step 4: Preparation of 3-((5-(4-(3-(benzoyloxy)propyl)piperidin-1-yl)-2-chlorophenyl)amino)propanoic acid

[0262] Benzoic acid 3-(1-(3-amino-4-chlorophenyl)piperidin-4-yl)propyl ester (900 mg, 2.42 mmol) and acrylic acid (1.74 g, 24.19 mmol) were added to H2O / AcOH (5 / 1, 12 mL), and the reaction solution was stirred at 110°C overnight under a nitrogen atmosphere. The reaction solution was directly concentrated under reduced pressure, and the crude product was purified by pre-HPLC (acetonitrile / formic acid water system) to obtain 3-((5-(4-(3-(benzoyloxy)propyl)piperidin-1-yl)-2-chlorophenyl)amino)propanoic acid.

[0263] LC-MS:(ESI,m / z):[M+H] + =445.2.

[0264] Step 5: Preparation of 3-(1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)phenyl)piperidin-4-yl)propyl benzoate

[0265] 3-((5-(4-(3-(Benzoyloxy)propyl)piperidin-1-yl)-2-chlorophenyl)amino)propanoic acid (850 mg, 1.91 mmol) was dissolved in AcOH (8 mL), followed by the addition of urea (1.19 g, 19.14 mmol). The reaction mixture was heated to 110°C and stirred overnight. Water (80 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (50 mL x 3). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 2:1) to obtain 3-(1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)phenyl)piperidin-4-yl)propyl benzoate.

[0266] LC-MS:(ESI,m / z):[M+H] + =470.1.

[0267] Step 6: Preparation of 1-(2-chloro-5-(4-(3-hydroxypropyl)piperidin-1-yl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0268] 3-(1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)phenyl)piperidin-4-yl)propyl benzoate (500 mg, 1.06 mmol) was dissolved in methanol / concentrated hydrochloric acid (1:1, 8 mL). The reaction mixture was heated to 70°C under a nitrogen atmosphere and stirred overnight. The mixture was concentrated under reduced pressure, and the crude product was purified by pre-HPLC (acetonitrile / formic acid / water system) to afford 1-(2-chloro-5-(4-(3-hydroxypropyl)piperidin-1-yl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione.

[0269] LC-MS:(ESI,m / z):[M+H] + =366.1.

[0270] Step 7: Preparation of 3-(1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)phenyl)piperidin-4-yl)propanal

[0271] 1-(2-Chloro-5-(4-(3-hydroxypropyl)piperidin-1-yl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione (200 mg, 0.55 mmol) was dissolved in dichloromethane (10 mL), Dess-Martin (466 mg, 1.10 mmol) was added and stirred at room temperature overnight. The reaction solution was directly concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (dichloromethane:methanol=10:1) to give 3-(1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)phenyl)piperidin-4-yl)propanal.

[0272] LC-MS:(ESI,m / z):[M+H] + =364.0.

[0273] Step 8: Preparation of tert-butyl 4-(3-(1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)phenyl)piperidin-4-yl)propyl)piperazine-1-carboxylate

[0274] 3-(1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)phenyl)piperidin-4-yl)propanal (110 mg, 0.30 mmol) and tert-butyl piperazine-1-carboxylate (84 mg, 0.45 mmol) were dissolved in THF (10 mL). The reaction mixture was stirred at room temperature for 1 hour. Sodium triacetoxyborohydride (95 mg, 0.45 mmol) was added and stirred at room temperature overnight. Water (50 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 2:1) to obtain tert-butyl 4-(3-(1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)phenyl)piperidin-4-yl)propyl)piperazine-1-carboxylate.

[0275] LC-MS:(ESI,m / z):[M+H] + =534.2.

[0276] Step 9: Preparation of 1-(2-chloro-5-(4-(3-(piperazin-1-yl)propyl)piperidin-1-yl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0277] Tert-butyl 4-(3-(1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)phenyl)piperidin-4-yl)propyl)piperazine-1-carboxylate (120 mg, 0.225 mmol) was dissolved in DCM / TFA (1:1, 8 mL) and stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure to afford 1-(2-chloro-5-(4-(3-(piperazin-1-yl)propyl)piperidin-1-yl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione. This product was used directly in the next step without purification.

[0278] LC-MS:(ESI,m / z):[M+H] + =434.2.

[0279] Step 10: Preparation of tert-butyl (3-((R)-1-((6-((1R,4R)-4-(4-(3-(1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)phenyl)piperidin-4-yl)propyl)piperazine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)carboxylate

[0280] 1-(2-chloro-5-(4-(3-(piperazin-1-yl)propyl)piperidin-1-yl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione (90 mg, 0.21 mmol) and (1R,4R)-4-(4-(((R)-1-(3-((tert-butoxycarbonyl)amino)-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylic acid (130 mg, 0.21 mmol) were dissolved in DMF (5 mL), and then DIEA (55 mg, 0.42 mmol) and HATU (120 mg, 0.32 mmol) were added and stirred at room temperature for 1 hour. The reaction solution was diluted with water (50 mL) and then washed with dichloromethane (50 mL x 4 ). 3) extraction, drying over anhydrous sodium sulfate, filtration, and concentration under reduced pressure. The crude product was purified by silica gel column chromatography (dichloromethane:methanol=10:1) to give tert-butyl (3-((R)-1-((6-((1R,4R)-4-(4-(3-(1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)phenyl)piperidin-4-yl)propyl)piperazine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)carboxylate.

[0281] LC-MS:(ESI,m / z):[M+H] + =1018.4.

[0282] Step 11: Preparation of 1-(5-(4-(3-(4-((1R,4R)-4-(4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperazin-1-yl)propyl)piperidin-1-yl)-2-chlorophenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0283] Tert-butyl (3-((R)-1-((6-((1R,4R)-4-(4-(3-(1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)phenyl)piperidin-4-yl)propyl)piperazine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)carboxylate (100 mg, 0.10 mmol) was dissolved in DCM / TFA (1 / 1, 8 mL). The mixture was stirred at room temperature for 1 hour, and the reaction solution was directly concentrated under reduced pressure. The crude product was purified by pre-HPLC to give 1-(5-(4-(3-(4-((1R,4R)-4-(4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperazin-1-yl)propyl)piperidin-1-yl)-2-chlorophenyl)dihydropyrimidine-2,4(1H,3H)-dione.

[0284] LC-MS:(ESI,m / z):[M+H] + =918.4.

[0285] 1 H NMR (400MHz, DMSO-d6) δ10.39 (s, 1H), 8.20-8.00 (m, 2H), 7.30 (d, J = 8.8Hz, 1H), 7.0 3(s,1H),6.99(s,1H),6.94-6.86(m,2H),6.85(s,1H),6.70(s,1H),5.63-5.46(m,3H ),3.88(s,3H),3.75-3.60(m,3H),3.60-3.53(m,1H),3.52-3.40(m,4H),2.98-2.92( m,1H),2.80-2.60(m,6H),2.35-2.28(m,8H),1.89-1.43(m,16H),1.29-1.11(m,4H).

[0286] Example 8: 1-(2-chloro-5-(4-(3-((1-((1R,4R)-4-(4-(((R)-1-(4-(2-chloro-6-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperidin-4-yl)oxy)propyl)piperazine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0287] Step 1: Preparation of benzyl 4-(3-ethoxy-3-oxopropoxy)piperidine-1-carboxylate

[0288] Benzyl 4-cyanopiperidine-1-carboxylate (5.0 g, 21.28 mmol) was dissolved in acetonitrile (100 mL), followed by the addition of methyl acrylate (4.256 g, 42.56 mmol) and cesium carbonate (13.87 g, 42.56 mmol). The reaction mixture was stirred at room temperature overnight. The mixture was concentrated under reduced pressure, and the residue was added with water (100 mL). The mixture was then extracted with ethyl acetate (100 mL x 3). The organic phase was washed with saturated brine (150 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 1:4) to afford benzyl 4-(3-ethoxy-3-oxopropoxy)piperidine-1-carboxylate.

[0289] LC-MS:(ESI,m / z):[M+H] + =336.1.

[0290] Step 2: Preparation of benzyl 4-(3-hydroxypropyloxy)piperidine-1-carboxylate

[0291] Benzyl 4-(3-ethoxy-3-oxopropoxy)piperidine-1-carboxylate (1.5 g, 4.47 mmol) was dissolved in anhydrous tetrahydrofuran (50 mL). Under a nitrogen atmosphere, borane in tetrahydrofuran (45 mL, 45 mmol) was slowly added to the solution. The reaction mixture was heated to 50°C and stirred for 3 hours. The mixture was cooled to room temperature, and methanol was slowly added dropwise until no bubbles were generated. The mixture was then concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (methanol:dichloromethane = 1:9) to obtain benzyl 4-(3-hydroxypropoxy)piperidine-1-carboxylate.

[0292] LC-MS:(ESI,m / z):[M+H] + =294.1.

[0293] Step 3: Preparation of benzyl 4-(3-oxopropoxy)piperidine-1-carboxylate

[0294] Benzyl 4-(3-hydroxypropoxy)piperidine-1-carboxylate (500 mg, 1.70 mmol) was dissolved in acetonitrile (15 mL), and IBX (952 mg, 3.40 mmol) was added. The reaction mixture was heated at reflux for 2 hours. The mixture was then concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (methanol:dichloromethane = 1:9) to obtain benzyl 4-(3-oxopropoxy)piperidine-1-carboxylate.

[0295] LC-MS:(ESI,m / z):[M+H] + =292.1.

[0296] Step 4: Preparation of tert-butyl 4-(3-((1-((benzyloxy)carbonyl)piperidin-4-yl)oxy)propyl)piperazine-1-carboxylate

[0297] Benzyl 4-(3-oxopropoxy)piperidine-1-carboxylate (370 mg, 1.27 mmol) was dissolved in THF (10 mL), and tert-butyl piperazine-1-carboxylate (236 mg, 1.27 mmol) was added sequentially and stirred at room temperature for 10 minutes. Sodium triacetoxyborohydride (808 mg, 3.81 mmol) was then added. The reaction mixture was stirred at room temperature for 2 hours. The mixture was diluted with water (50 mL), extracted with dichloromethane (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (methanol:dichloromethane = 1:9) to obtain tert-butyl 4-(3-((1-((benzyloxy)carbonyl)piperidin-4-yl)oxy)propyl)piperazine-1-carboxylate.

[0298] LC-MS:(ESI,m / z):[M+H] + =462.2.

[0299] Step 5: Preparation of tert-butyl 4-(3-(piperidin-4-yloxy)propyl)piperazine-1-carboxylate

[0300] tert-Butyl 4-(3-((1-((benzyloxy)carbonyl)piperidin-4-yl)oxy)propyl)piperazine-1-carboxylate (250 mg, 0.54 mmol) was dissolved in ethyl acetate (10 mL). Pd / C (10%, 75 mg) was added. The reaction system was replaced with hydrogen three times, then heated to 70°C under a hydrogen atmosphere and stirred overnight. The mixture was filtered, and the filter cake was rinsed with ethyl acetate (5 mL x 3). The filtrate was then concentrated under reduced pressure to yield tert-butyl 4-(3-(piperidin-4-yloxy)propyl)piperazine-1-carboxylate.

[0301] LC-MS:(ESI,m / z):[M+H] + =328.3.

[0302] Step 6: Preparation of tert-butyl 4-(3-((1-((1R,4R)-4-(4-(((R)-1-(4-(2-chloro-6-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperidin-4-yl)oxy)propyl)piperazine-1-carboxylate

[0303] Tert-butyl 4-(3-(piperidin-4-yloxy)propyl)piperazine-1-carboxylate (110 mg, 0.34 mmol) was dissolved in DMF (5 mL), and (1R,4R)-4-(4-(((R)-1-(4-(2-chloro-6-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylic acid (202 mg, 0.34 mmol), HATU (144 mg, 0.44 mmol) and DIEA (132 mg, 1.02 mmol) were added respectively, and the reaction solution was stirred at room temperature overnight. The mixture was diluted with water (50 mL), extracted with dichloromethane (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (methanol:dichloromethane = 1:9) to give tert-butyl 4-(3-((1-((1R,4R)-4-(4-(((R)-1-(4-(2-chloro-6-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperidin-4-yl)oxy)propyl)piperazine-1-carboxylate.

[0304] LC-MS:(ESI,m / z):[M+H] + =902.4.

[0305] Step 7: Preparation of ((1R,4R)-4-(4-(((R)-1-(4-(2-chloro-6-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexyl)(4-(3-(piperazin-1-yl)propoxy)piperidin-1-yl)methanone

[0306] Tert-butyl 4-(3-((1-((1R,4R)-4-(4-(((R)-1-(4-(2-chloro-6-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperidin-4-yl)oxy)propyl)piperazine-1-carboxylate (90 mg, 0.10 mmol) was dissolved in dichloromethane (5 mL) and trichloromethane was added. Fluoroacetic acid (1 mL) was added and stirred at room temperature for 2 hours. The reaction solution was directly concentrated under reduced pressure to give ((1R,4R)-4-(4-(((R)-1-(4-(2-chloro-6-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexyl)(4-(3-(piperazin-1-yl)propoxy)piperidin-1-yl)methanone, which was used directly in the next step without purification;

[0307] LC-MS:(ESI,m / z):[M+H]+ =802.4.

[0308] Step 8: Preparation of 1-(2-chloro-5-(4-(3-((1-((1R,4R)-4-(4-(((R)-1-(4-(2-chloro-6-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperidin-4-yl)oxy)propyl)piperazine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0309] ((1R,4R)-4-(4-(((R)-1-(4-(2-chloro-6-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexyl)(4-(3-(piperazin-1-yl)propoxy)piperidin-1-yl)methanone (90 mg, 0.10 mmol) was dissolved in dimethyl sulfoxide (5 mL), and pentafluorophenyl 4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoate (44 mg, 0.10 mmol) and DIEA (129 mg, 1.0 mmol) were added, and the reaction solution was stirred at room temperature for 2 hours. The reaction mixture was diluted with water (50 mL), extracted with dichloromethane (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by pre-HPLC to give 1-(2-chloro-5-(4-(3-((1-((1R,4R)-4-(4-(((R)-1-(4-(2-chloro-6-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperidin-4-yl)oxy)propyl)piperazine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione.

[0310] LC-MS:(ESI,m / z):[M+H] + =1054.5.

[0311] 1H NMR (400MHz, DMSO-d6) δ10.51(s,1H),8.31(d,J=8.2Hz,1H),8.05(s,1H),7.65(s,1H),7.63(s,1H),7.56( s,1H),7.46-7.31(m,4H),7.24(s,1H),7.02(s,1H),6.96(s,1H),6.05-5.86(m,1H),3.89(s,3H),3.87-3.8 0(m,1H),3.79-3.56(m,6H),3.52-3.42(m,4H),3.16-3.06(m,3H),2.98-2.89(m,1H),2.77-2.60(m,5H),2 .44-2.32(m,8H),2.01(s,6H),1.89-1.75(m,6H),1.74-1.68(m,3H),1.68-1.48(m,6H),1.44-1.26(m,2H).

[0312] Example 9: 3-((R)-1-((6-((1R,4R)-4-(4-((3-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)-3-azaspiro[5.5]undec-9-yl)methyl)piperazine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)-2-methylbenzylcyanide

[0313] Step 1: Preparation of tert-butyl 9-((4-((benzyloxy)carbonyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carboxylate

[0314] tert-Butyl 9-formyl-3-azaspiro[5.5]undecane-3-carboxylate (300 mg, 1.06 mmol) and benzyl piperazine-1-carboxylate (282 mg, 1.27 mmol) were dissolved in tetrahydrofuran (10 mL). Sodium triacetoxyborohydride (674 mg, 3.18 mmol) was added with stirring and the mixture was stirred at room temperature overnight. Water (50 mL) was added to the reaction solution, which was extracted with dichloromethane (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 1:1) to obtain tert-butyl 9-((4-((benzyloxy)carbonyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carboxylate.

[0315] LC-MS:(ESI,m / z):[M+H] + =486.3.

[0316] Step 2: Preparation of tert-butyl 9-(piperazin-1-ylmethyl)-3-azaspiro[5.5]undecane-3-carboxylate

[0317] tert-Butyl 9-((4-((benzyloxy)carbonyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carboxylate (200 mg, 0.41 mmol) was dissolved in ethyl acetate (15 mL), and palladium hydroxide on carbon (20%, 100 mg) was added. The resulting mixture was heated to 70°C under a hydrogen atmosphere and stirred overnight. After cooling to room temperature, the reaction solution was filtered through celite, and the filtrate was directly concentrated under reduced pressure to obtain tert-butyl 9-(piperazin-1-ylmethyl)-3-azaspiro[5.5]undecane-3-carboxylate, which was used directly in the next reaction without purification.

[0318] LC-MS:(ESI,m / z):[M+H] + =352.4.

[0319] Step 3: Preparation of methyl (1R,4R)-4-(4-(((R)-1-(3-cyano-2-methylphenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylate

[0320] Methyl (1R,4R)-4-(4-hydroxy-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylate (200 mg, 0.61 mmol), BOP (344 mg, 0.78 mmol) and DBU (232 mg, 1.52 mmol) were dissolved in DMF (5 mL) and stirred at room temperature for 30 minutes. Then, (R)-3-(1-aminoethyl)-2-methylbenzonitrile (154 mg, 0.78 mmol) was added, and the reaction solution was heated to 70°C and stirred overnight. The reaction mixture was cooled to room temperature, diluted with water (50 mL), extracted with ethyl acetate (30 mL x 3), and the organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 9:1) to give (1R,4R)-4-(4-(((R)-1-(3-cyano-2-methylphenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylic acid methyl ester.

[0321] LC-MS:(ESI,m / z):[M+H] + =473.2.

[0322] Step 4: Preparation of (1R,4R)-4-(4-(((R)-1-(3-cyano-2-methylphenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylic acid

[0323] Methyl (1R,4R)-4-(4-(((R)-1-(3-cyano-2-methylphenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylate (130 mg, 0.27 mmol) was dissolved in a mixture of tetrahydrofuran / water (10 mL / 5 mL). Lithium hydroxide (53 mg, 2.2 mmol) was added, and the reaction mixture was heated to 50°C and stirred overnight. The mixture was cooled to room temperature, diluted with water (30 mL), and the pH of the mixture was adjusted to neutral with saturated aqueous sodium citrate solution. The mixture was extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain (1R,4R)-4-(4-(((R)-1-(3-cyano-2-methylphenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylic acid.

[0324] LC-MS:(ESI,m / z):[M+H] + =459.2.

[0325] Step 5: Preparation of tert-butyl 9-((4-((1R,4R)-4-(4-(((R)-1-(3-cyano-2-methylphenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carboxylate

[0326] (1R,4R)-4-(4-(((R)-1-(3-cyano-2-methylphenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylic acid (120 mg, 0.26 mmol) and tert-butyl 9-(piperazin-1-ylmethyl)-3-azaspiro[5.5]undecane-3-carboxylate (120 mg, 0.34 mmol) were dissolved in DMF (5 mL), and HATU (148 mg, 0.39 mmol) and N,N-diisopropylethylamine (100 mg, 0.78 mmol) were added, and the reaction solution was stirred at room temperature for 2 hours. The reaction solution was diluted with water (50 mL) and extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (methanol:dichloromethane = 1:9) to give tert-butyl 9-((4-((1R,4R)-4-(4-(((R)-1-(3-cyano-2-methylphenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carboxylate.

[0327] LC-MS:(ESI,m / z):[M+H] + =792.4.

[0328] Step 6: Preparation of 3-((R)-1-((6-((1R,4R)-4-(4-((3-azaspiro[5.5]undec-9-yl)methyl)piperazine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)-2-methylbenzonitrile

[0329] tert-Butyl 9-((4-((1R,4R)-4-(4-(((R)-1-(3-cyano-2-methylphenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carboxylate (160 mg, 0.20 mmol) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (2 mL) was added and stirred at room temperature for 2 hours. The reaction solution was directly concentrated under reduced pressure to give 3-((R)-1-((6-((1R,4R)-4-(4-((3-azaspiro[5.5]undec-9-yl)methyl)piperazine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)-2-methylbenzonitrile, which was used directly in the next reaction without purification.

[0330] LC-MS:(ESI,m / z):[M+H] + =692.4.

[0331] Step 7: Preparation of 3-((R)-1-((6-((1R,4R)-4-(4-((3-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)-3-azaspiro[5.5]undec-9-yl)methyl)piperazine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)-2-methylbenzonitrile

[0332] 3-((R)-1-((6-((1R,4R)-4-(4-((3-azaspiro[5.5]undec-9-yl)methyl)piperazine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)-2-methylbenzonitrile (160 mg, TFA salt, 0.20 mmol) was dissolved in dimethyl sulfoxide (3 mL), and pentafluorophenyl 4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoate (95 mg, 0.22 mmol) and N,N-diisopropylethylamine (77 mg, 0.60 mmol) were added with stirring. The reaction solution was stirred at room temperature overnight, water (50 mL) was added to the reaction solution, and the reaction solution was filtered. The filter cake was purified by pre-HPLC (CH3CN / 0.08% NH4HCO3 aqueous solution, 5% to 95%) to give 3-((R)-1-((6-((1R,4R)-4-(4-((3-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)-3-azaspiro[5.5]undec-9-yl)methyl)piperazine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)-2-methylbenzonitrile.

[0333] LC-MS:(ESI,m / z):[M+H] + =942.5.

[0334] 1 H NMR(400MHz,DMSO-d6)δ10.52(s,1H),8.25(s,1H),7.83(d,J=8.0Hz,1H),7.71-7 .60(m,2H),7.55(s,1H),7.39(t,J=7.7Hz,2H),7.02(s,1H),5.78-5.68(m,1H),3. 91(s,3H),3.81-3.38(m,8H),3.3-3.25(m,2H),3.01-2.93(m,1H),2.80-2.63(m, 7H),2.48-2.21(m,7H),1.90-1.77(m,4H),1.75-1.20(m,18H),1.16-0.92(m,4H).

[0335] Example 10: 1-(2-chloro-5-(4-((2-(1-((1R,4R)-4-(4-(((R)-1-(4-(2-chloro-6-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperidin-4-yl)ethoxy)methyl-d2)piperidin-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0336] 1-(2-chloro-5-(4-((2-(piperidin-4-yl)ethoxy)methyl-d2)piperidin-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione (190 mg, 0.397 mmol) was dissolved in DMF (5 mL), and (1R,4R)-4-(4-(((R)-1-(4-(2-chloro-6-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylic acid (85 mg, 0.143 mmol), DIEA (156 mg, 1.2 mmol) and PyBOP (250 mg, 0.48 mmol) were added respectively. The reaction solution was stirred at 60°C under N2 protection for 16 h. The reaction solution was diluted with water (40 mL) and washed with ethyl acetate (40 mL x 40 mL). 4), extracted with saturated brine, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by pre-HPLC to give 1-(2-chloro-5-(4-((2-(1-((1R,4R)-4-(4-(((R)-1-(4-(2-chloro-6-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperidin-4-yl)ethoxy)methyl-d2)piperidine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione.

[0337] LC-MS:(ESI,m / z):[M+H] + =1053.4.

[0338] 1H NMR(400MHz,MeOD)δ8.01(s,1H),7.64(s,1H),7.62(s,1H),7.52(s,1H),7.44-7.35(m,3H),7.30(t,J=7.8Hz,1H) ,7.09(d,J=1.1Hz,1H),7.01(s,1H),6.95(s,1H),6.06(d,J=6.8Hz,1H),4.68-4.47(m,2H),4.10-4.00(m,1H),3. 95(s,3H),3.82-3.60(m,4H),3.49(t,J=6.2Hz,2H),3.27(s,2H),3.17-3.02(m,3H),2.91-2.80(m,3H),2.75-2.6 7(m,1H),2.66-2.55(m,1H),2.52(s,3H),2.05(s,6H),2.00-1.58(m,19H),1.58-1.49(m,2H),1.32-1.29(m,2H).

[0339] Example 11: 1-(5-(4-((2-(1-((1R,4R)-4-(4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperidin-4-yl)ethoxy)methyl-d2)piperidine-1-carbonyl)-2-chlorophenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0340] Step 1: Preparation of (1R,4R)-4-(4-(((R)-1-(3-((tert-butoxycarbonyl)amino)-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylic acid pentafluorophenyl ester

[0341] (1R,4R)-4-(4-(((R)-1-(3-((tert-butoxycarbonyl)amino)-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylic acid (200 mg, 0.33 mmol) was dissolved in DMF (5 mL), 2,3,4,5,6-pentafluorophenol (74 mg, 0.40 mmol) and DCC (82 mg, 0.40 mmol) were added, and the reaction mixture was heated to 70°C and stirred for 2 h. The reaction mixture was diluted with water (50 mL) and extracted with dichloromethane (50 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (ethyl acetate:petroleum ether=1:1) to give pentafluorophenyl (1R,4R)-4-(4-(((R)-1-(3-((tert-butoxycarbonyl)amino)-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylate.

[0342] LC-MS:(ESI,m / z):[M+H] + =769.4.

[0343] Step 2: Preparation of tert-butyl (3-((R)-1-((6-((1R,4R)-4-(4-(2-((1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)piperidin-4-yl)methoxy-d2)ethyl)piperidine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)carboxylate

[0344] Pentafluorophenyl (1R,4R)-4-(4-(((R)-1-(3-((tert-butoxycarbonyl)amino)-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylate (200 mg, 0.26 mmol) was dissolved in dimethyl sulfoxide (5 mL), and 1-(2-chloro-5-(4-((2-(piperidin-4-yl)ethoxy)methyl-d2)piperidine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione (144 mg, 0.30 mmol) and DIEA (117 mg, 0.90 mmol) were added, and the reaction solution was stirred at room temperature for 2 hours. The reaction solution was diluted with water (50 mL), extracted with dichloromethane (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (methanol: dichloromethane = 1:9) to give tert-butyl (3-((R)-1-((6-((1R,4R)-4-(4-(2-((1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)piperidin-4-yl)methoxy-d2)ethyl)piperidine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)carboxylate.

[0345] LC-MS:(ESI,m / z):[M+H] + =1063.4.

[0346] Step 3: Preparation of 1-(5-(4-((2-(1-((1R,4R)-4-(4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperidin-4-yl)ethoxy)methyl-d2)piperidine-1-carbonyl)-2-chlorophenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0347] Tert-butyl (3-((R)-1-((6-((1R,4R)-4-(4-(2-((1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)piperidin-4-yl)methoxy-d2)ethyl)piperidine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)carboxylate (150 mg, 0.14 mmol) was dissolved in DCM (5 mL), TFA (1 mL) was added, and the mixture was stirred at room temperature for 1 hour. After concentration under reduced pressure, MeOH / DCM (1 / 9, 10 mL) was added to dissolve the mixture, and a saturated sodium bicarbonate aqueous solution diluted one-half was added to adjust the pH to neutral. After stirring for 10 minutes, the mixture was washed with MeOH / DCM (1 / 9, 30 mL x 4 ). 3) extraction, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by pre-HPLC to give 1-(5-(4-((2-(1-((1R,4R)-4-(4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperidin-4-yl)ethoxy)methyl-d2)piperidine-1-carbonyl)-2-chlorophenyl)dihydropyrimidine-2,4(1H,3H)-dione.

[0348] LC-MS:(ESI,m / z):[M+H] + =963.2

[0349] 1 H NMR (400MHz, DMSO-d6) δ10.52(s,1H),8.12(d,J=8.1Hz,1H),8.06(s,1H),7.63(d,J=8.2Hz,1H), 7.54(d,J=2.0Hz,1H),7.38(dd,J=8.2,2.0Hz,1H),6.99(s,1H),6.88(s,1H),6.85(s,1H),6.70(s ,1H),5.62-5.51(m,3H),4.54-4.30(m,2H),3.96-3.83(m,4H),3.80-3.51(m,3H),3.40(t,J=6.4H z,2H),3.11-2.89(m,3H),2.80-2.59(m,4H),2.36(s,3H),1.92-1.37(m,20H),1.20-0.89(m,4H).

[0350] Example 12: 1-(2-chloro-5-(9-((4-((1R,4R)-4-(4-(((R)-1-(3-(difluoromethyl)-2-methylphenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0351] Step 1: Preparation of methyl (1R,4R)-4-(4-(((R)-1-(3-(difluoromethyl)-2-methylphenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylate

[0352] Methyl (1R,4R)-4-(4-hydroxy-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylate (400 mg, 1.212 mmol) was dissolved in DMF (5 mL), and BOP (696 mg, 1.576 mmol) and DBU (461 mg, 3.03 mmol) were added. After stirring at room temperature for 0.5 hour, (R)-1-(3-(difluoromethyl)-2-methylphenyl)ethyl-1-amine (244 mg, 1.33 mmol) was added, and the reaction solution was heated to 70°C and stirred overnight. The reaction solution was diluted with water (50 mL) and extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated aqueous sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 9:1) to give (1R,4R)-4-(4-(((R)-1-(3-(difluoromethyl)-2-methylphenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylic acid methyl ester.

[0353] LC-MS:(ESI,m / z):[M+H] + =498.2.

[0354] Step 2: Preparation of (1R,4R)-4-(4-(((R)-1-(3-(difluoromethyl)-2-methylphenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylic acid

[0355] Methyl (1R,4R)-4-(4-(((R)-1-(3-(difluoromethyl)-2-methylphenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylate (178 mg, 0.36 mmol) was dissolved in THF (5 mL), and LiOH (44 mg, 1.83 mmol) was added. The reaction mixture was heated to 50°C and stirred for 2 hours. The mixture was diluted with water (50 mL), and the pH was adjusted to neutral with citric acid. The mixture was extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford (1R,4R)-4-(4-(((R)-1-(3-(difluoromethyl)-2-methylphenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylic acid, which was used directly in the next reaction without purification.

[0356] LC-MS:(ESI,m / z):[M+H] + =484.2.

[0357] Step 3: Preparation of 1-(2-chloro-5-(9-((4-((1R,4R)-4-(4-(((R)-1-(3-(difluoromethyl)-2-methylphenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0358] (1R,4R)-4-(4-(((R)-1-(3-(difluoromethyl)-2-methylphenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylic acid (128 mg, 0.265 mmol) was dissolved in DMF (5 mL), and 1-(2-chloro-5-(9-(piperazin-1-ylmethyl)-3-azaspiro[5.5]undecane-3-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione (133 mg, 0.265 mmol), DIEA (103 mg, 0.795 mmol) and HATU (141 mg, 0.307 mmol) were added respectively, and the reaction solution was stirred at room temperature for 2 hours. The mixture was diluted with water (50 mL) and extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated aqueous sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by Prep-HPLC (CH3CN / 0.08% aqueous NH4HCO3, 5% to 95%) to give 1-(2-chloro-5-(9-((4-((1R,4R)-4-(4-(((R)-1-(3-(difluoromethyl)-2-methylphenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione.

[0359] LC-MS:(ESI,m / z):[M+H] + =967.3.

[0360] 1 H NMR (400MHz, DMSO-d6) δ10.51(s,1H),8.23(d,J=7.3Hz,1H),8.08(s,1H),7.68(d,J=8.0Hz,1H),7.63(d,J=8.2Hz ,1H),7.55(d,J=1.9Hz,1H),7.41-7.35(m,2H),7.35-7.04(m,2H),6.97(s,1H),5.82-5.72(m,1H),3.87(s,3H),3. 80-3.69(m,1H),3.66-3.54(m,3H),3.54-3.42(m,4H),3.30-3.20(m,2H),3.00-2.89(m,1H),2.78-2.70(m,2H),2 .58-2.54(m,3H),2.38-2.21(m,7H),2.15-2.07(m,2H),1.91-1.77(m,4H),1.74-1.24(m,17H),1.17-0.95(m,4H).

[0361] Example 13: 1-(5-(4-(3-(4-((1R,4R)-4-(4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperazin-1-yl)propyloxy)piperidine-1-carbonyl)-2-chlorophenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0362] Step 1: Preparation of tert-butyl (3-((R)-1-((6-((1R,4R)-4-(4-(3-((1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)piperidin-4-yl)oxy)propyl)piperazine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)carboxylate

[0363] 1-(2-Chloro-5-(4-(3-(piperazin-1-yl)propoxy)piperidine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione (90 mg, 0.13 mmol), HATU (59 mg. 0.16 mmol) and DIEA (50 mg, 0.39 mmol) were added to (1R,4R)-4-(4-(((R)-1-(3-((tert-butoxycarbonyl)amino)-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carboxylic acid (80 mg, 0.16 mmol) The mixture was added into a DMF (5 mL) solution and stirred at room temperature for 2 hours. Water (50 mL) was added and stirred. The mixture was filtered. The filter cake was washed 3 times with water and then dissolved in dichloromethane (20 mL). It was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude product of tert-butyl (3-((R)-1-((6-((1R,4R)-4-(4-(3-((1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)piperidin-4-yl)oxy)propyl)piperazine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)carboxylate.

[0364] LC-MS:(ESI,m / z):[M+H] + =1062.5.

[0365] Step 2: Preparation of 1-(5-(4-(3-(4-((1R,4R)-4-(4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperazin-1-yl)propoxy)piperidine-1-carbonyl)-2-chlorophenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0366] Tert-butyl (3-((R)-1-((6-((1R,4R)-4-(4-(3-((1-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)piperidin-4-yl)oxy)propyl)piperazine-1-carbonyl)cyclohexyl)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)carboxylate (130 mg, 0.12 mmol) was dissolved in dichloromethane (5 mL) and trifluoroethane was added. The reaction mixture was stirred at room temperature for 2 hours, concentrated under reduced pressure, and the crude product was purified by reverse phase preparative method to give 1-(5-(4-(3-(4-((1R,4R)-4-(4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-7-methoxy-2-methylquinazolin-6-yl)cyclohexane-1-carbonyl)piperazin-1-yl)propoxy)piperidine-1-carbonyl)-2-chlorophenyl)dihydropyrimidine-2,4(1H,3H)-dione.

[0367] LC-MS:(ESI,m / z):[M+H] + =962.5.

[0368] 1 H NMR(400MHz,DMSO-d6)δ10.51(s,1H),8.17–7.98(m,2H),7.67–7.50(m,2H),7.47–7.3 4(m,1H),6.99(s,1H),6.88(s,1H),6.85(s,1H),6.70(s,1H),5.64–5.46(m,3H),3.88 (s,4H),3.74(s,1H),3.62(d,J=5.7Hz,1H),3.56–3.39(m,8H),3.18(s,1H),3.00-2.8 9(m,1H),2.80–2.70(m,2H),2.68-2.59(m,1H),2.40–2.25(m,9H),1.94–1.35(m,18H).

[0369] II. Biological Activity Test Example

[0370] Test Example 1: KRAS-G12C / SOS1 binding assay

[0371] The following method is used to determine the extent of KRAS-G12C / SOS1 binding inhibition by preferred compounds of the present invention under in vitro conditions. This method uses a KRAS-G12C / SOS1 Binding Assay Kit (Cisbio, 63ADK000CB16PEG) and homogeneous time-resolved fluorescence (HTRF) to determine competitive inhibition of KRAS-G12C / SOS1 binding by preferred compounds.

[0372] The experimental procedure is as follows (refer to the kit instructions for detailed procedures): The compound of the present invention was first dissolved in DMSO to a concentration of 20 mM. The test compound was then diluted isocratically with the buffer provided in the kit to a final concentration range of 10,000 nM to 0.04 nM in the reaction system. The final DMSO concentration was 0.5%. 2 μL of the compound was incubated with 4 μL of 1x Tag1-SOS1 and 4 μL of 1x Tag2-KRAS-G12C (containing 10 μM guanosine triphosphate (GTP)) at 25°C for 15 minutes. Subsequently, 5 μL of 1x Anti-Tag1-Tb3+ and 5 μL of 1x Anti-Tag2-XL665 were added to the reaction system and incubated on ice for 3 hours. After incubation, the fluorescence intensity of each well was measured at an excitation wavelength of 337 nm on an EnVision microplate reader (PerkinElmer, 2105) in HTRF mode. The fluorescence intensity of each well was read at emission wavelengths of 620 nm and 665 nm. The Ratio value was calculated using the formula Ratio = (665 nm / 620 nm) × 104. The inhibition rate of the compound at each concentration was calculated by comparing the fluorescence intensity ratio with that of the control group. Nonlinear curve fitting using the logarithmic concentration-inhibition rate was then performed using GraphPad Prism 8 to determine the IC50 value of the compound (see Table 1).

[0373] Test Example 2: SOS1 degradation test

[0374] The degradation effect of the compounds on SOS1 was investigated in human non-small cell lung cancer cells NCI-H358 (ATCC or Chinese Academy of Sciences Stem Cell Bank) with KRAS G12C mutation, human lung cancer cells A-427 (ATCC) with KRAS G12D mutation, human colorectal adenocarcinoma cells SW-620 (ATCC) with KRAS G12V mutation, and human colorectal adenocarcinoma epithelial cells DLD-1 (ATCC) with KRAS G13D mutation. The specific method is as follows:

[0375] Plate 0.95 mL of cells per well of a 24-well cell culture plate to a cell density of 5 × 105 cells / well. Incubate the plate in a 5% CO2 incubator at 37°C overnight. Add 50 μL of the diluted compound solution to the corresponding well containing cells, ensuring a final compound concentration ranging from 0.03 to 3000 nM. The final DMSO concentration is 0.25%. After dosing, the cell plates were incubated at 37°C in a 5% CO2 incubator for 24 hours. The cell culture medium in each well was removed and washed twice with 1× PBS. Adherent NCI-H358 cells were lysed in 180 μL of RIPA (Strong) lysis buffer (Biyuntian, P0013B) supplemented with 1 mM phenylmethylsulfonyl fluoride, protease inhibitor cocktail (Biyuntian, P1008), and protease phosphatase inhibitor cocktail (Biyuntian, P1045). After incubation on ice for 30 minutes, the protein lysate from each well was transferred to a 1.5 mL centrifuge tube and centrifuged at 15,000 g at 4°C for 20 minutes. The supernatant was frozen at -80°C and used for Western blotting to detect SOS1 protein degradation. Total protein concentration in the supernatant was determined using a BCA protein quantification kit (Tiangen, PA115-02).

[0376] Test Example 3: Human SOS1 protein immunoblotting assay

[0377] The total protein concentration in the cell lysate was determined by BCA assay. The protein concentration was adjusted to 0.5 μg / μL using PBS and 5× SDS-PAGE protein loading buffer (Biyuntian, P0015L). The mixture was incubated at 100°C for 15 minutes, placed on ice for 5 minutes, and centrifuged at 14,000g at 4°C for 1 minute. The mixture was then mixed and used as the sample for Western blotting. Protein electrophoresis was performed on a precast gel (Keygen, KGMG010W15) with a loading volume of 10 μL (5 μg total protein) and Tris-MOPS-SDS electrophoresis buffer (Adamas, P1598253). Electrophoresis was performed at a constant voltage of 120 V for 55 minutes. After electrophoresis, the proteins were transferred to a PVDF membrane at a constant current of 250 mA for 65 minutes. After transfer, the membrane was incubated in 1× QuickBlock blocking buffer (Biyuntian, P0235) at room temperature for 30 minutes. After blocking, the PVDF membranes were incubated with the primary antibody against SOS1 (Abcam, ab140621) at 4°C overnight. The membranes were then washed with TBST buffer (2.4 g Tris, 8.8 g NaCl, 1.5 mL Tween 20, adjusted to pH 7.4, and diluted to 1 L) for 30 minutes (10 minutes each wash). The membranes were then incubated with the secondary antibody (Abcam, ab205718) at room temperature for 2 hours. The membranes were then washed with TBST buffer for 30 minutes (10 minutes each wash). Finally, the membranes were incubated with Clarity Western ECL Substrate (BIO-RAD, 170-5061) for 5 minutes for luminescence development. The protein images were visualized and photographed using a chemiluminescence imaging system (ChemiScope 6200 Touch, Qinxiang). The grayscale values ​​of the protein images were analyzed using Qinxiang chemiluminescence analysis software. The grayscale correction value for each sample was calculated using the formula: grayscale correction value = (grayscale value of the target protein / grayscale value of the corresponding internal reference) × 10³. The degradation rate was then calculated by comparing it with the grayscale correction value of the control group. Then, the DC of the compound was obtained by nonlinear curve fitting using the logarithmic concentration-inhibition rate using GraphPad Prism 8. 50 and D max value.

[0378] Test Example 4: 3D cell proliferation inhibition test

[0379] The inhibitory effect of the compounds on 3D cell proliferation was investigated in human non-small cell lung cancer cells NCI-H358 (ATCC or Chinese Academy of Sciences Stem Cell Bank) with KRAS G12C mutation, human metastatic pancreatic adenocarcinoma cells AsPC-1 (ATCC) with KRAS G12D mutation, human lung adenocarcinoma cells NCI-H441 (ATCC) with KRAS G12V mutation, and human colorectal adenocarcinoma epithelial cells DLD-1 (ATCC) with KRAS G13D mutation. The specific method is as follows:

[0380] Using an Echo acoustic pipetting system (Labcyte, Echo 550), 200 nL of a serial dilution of the compound solution was added to the corresponding wells of a 384-well low-adhesion microplate (Corning, 3657) to achieve a final concentration range of 20,000 nM to 0.008 nM. 40 μL of cells at a given density were then added to the corresponding wells of the microplate. In addition to the wells containing the test compound, DMSO control wells and culture medium control wells were also set up. The DMSO control wells contained DMSO and cells, while the culture medium wells contained only culture medium. After sample addition, the plate was covered and the 384-well low-adhesion microplate was placed in a 5% CO2 incubator and incubated at 37°C for 7 days. After 7 days, the microplate was removed and 40 μL of CTG reagent (Promega, G9683) was added to each well. The cells were incubated at room temperature for 30 minutes before reading the chemiluminescence on an EnVision microplate reader. The percentage of cell proliferation inhibition was calculated using the formula: inhibition rate % = (mean value of DMSO control group - single concentration reading of compound) / (mean value of DMSO control group - mean value of culture medium control group) × 100. The inhibition rate of the compound at each concentration was calculated, and then a nonlinear curve fitting was performed using the logarithmic concentration-inhibition rate using GraphPad Prism 8 to obtain the IC value of the compound. 50 value.

[0381] The experimental results are shown in Table 1:

[0382] Table 1 “-” means not tested;

[0383] When DC 50 ≤50nM is A, when 50nM <DC 50 When ≤100nM, it is B. When 100nM <DC 50 ≤500nM is C, when 500nM <DC 50 ≤1000nM is D; 1000nM <DC 50 ≤10 μM is E;

[0384] When 80≤D max (%)≤100 is A, when 50≤D max (%) <80 is B, when 30≤D max (%) <50 is C, when D max (%) < 30, D;

[0385] When IC 50 ≤15nM is A, when 15nM <IC 50 When ≤30nM, it is B. When 30nM <IC 50 ≤50nM is C, when 50nM <IC 50≤100nM is D, when 100nM <IC 50 E is ≤1000nM.

[0386] Experimental conclusion: The compounds of the present invention can effectively bind to the SOS1 target protein or produce an inhibitory effect. The compounds of the present invention can effectively and specifically degrade the SOS1 protein.

[0387] Although the above describes specific embodiments of the present invention, it should be understood by those skilled in the art that these are merely illustrative and that various changes or modifications may be made to these embodiments without departing from the principles and essence of the present invention. Therefore, the scope of protection of the present invention is defined by the appended claims.

Claims

1. A compound of formula I, its stereoisomers, enantiomers, diastereomers, deuterated forms, hydrates, solvates, prodrugs, or pharmaceutically acceptable salts: SLE I in, S is L is L is connected to S through A or B; E is 2. The compound of formula I according to claim 1, its stereoisomers, enantiomers, diastereomers, deuterated forms, hydrates, solvates, prodrugs, or pharmaceutically acceptable salts thereof, characterized in that: The compound represented by formula I satisfies one or more of the following conditions: (1) S is L is (2) S is L is (3) S is L is (4) S is L is (5) S is L is (6) S is L is (7) S is L is (8) S is L is (9) S is L is (10)S is E is (11) S is E is (12) S is E is (13)S is E is (14) S is E is (15)S is E is (16)S is E is (17)S is E is (18)S is E is (19)S is E is (20) L is E is (21) L is E is (22) L is E is (23) L is E is (24) L is E is (25) L is E is (26) L is E is (27) L is E is (28) L is E is (29) L is E is (30) L is E is (31) L is E is (32) L is E is (33) L is E is (34) L is E is and (35) L is E is 3. The compound of formula I according to claim 1, its stereoisomers, enantiomers, diastereomers, deuterated forms, hydrates, solvates, prodrugs, or pharmaceutically acceptable salts thereof, characterized in that: The compound represented by formula I satisfies one or more of the following conditions: (1) S is L is E is (2) S is L is (3) S is L is E is (4) S is L is E is (5) S is L is E is (6) S is L is E is (7) S is L is E is (8) S is L is E is and (9) S is L is E is 4. The compound of formula I according to claim 3, its stereoisomers, enantiomers, diastereomers, deuterated forms, hydrates, solvates, prodrugs, or pharmaceutically acceptable salts thereof, characterized in that: In L, A and S are connected.

5. The compound of formula I according to claim 1, its stereoisomers, enantiomers, diastereomers, deuterated forms, hydrates, solvates, prodrugs, or pharmaceutically acceptable salts thereof, characterized in that: The compound represented by formula I is any of the following compounds:

6. A pharmaceutical composition comprising G and a pharmaceutically acceptable carrier, diluent or excipient, wherein G is a compound according to at least one of claims 1 to 5, a stereoisomer, enantiomer, diastereomer, deuterate, hydrate, solvate, prodrug, or a pharmaceutically acceptable salt thereof.

7. Use of a compound according to at least one of claims 1 to 5, a stereoisomer, enantiomer, diastereomer, deuterate, hydrate, solvate, metabolite, prodrug, or pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 6, for the preparation of a medicament for treating or preventing a disease or condition mediated by SOS1, or a disease or condition caused by the interaction of SOS1 with Ras or SOS1 with Rac, or cancer; preferably, the disease or condition mediated by SOS1 is cancer, neurofibromatosis type 1 (NF1), Noonan syndrome (NS), Noonan syndrome with multiple lentigines (NSML), capillary malformation-arteriovenous malformation syndrome (CM-AVM), Costello syndrome (CS), cardiofacial-cutaneous syndrome (CFC), Leggs syndrome, or hereditary gingival fibromatosis; preferably, the disease or condition caused by the interaction of SOS1 with Ras or SOS1 with Rac is cancer.

8. The use according to claim 7, characterized in that The cancer is selected from: Heart: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma, and teratoma group; Lung: bronchial cancer (squamous cell carcinoma, undifferentiated small cell carcinoma, undifferentiated large cell carcinoma, adenocarcinoma), alveolar carcinoma (bronchiolar carcinoma), bronchial adenoma, sarcoma, lymphoma, enchondroma, hamartoma, mesothelioma; Gastrointestinal tract: esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumor, vipoma), small intestine (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large intestine (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma); Genitourinary system: kidney (adenocarcinoma, Wilms' tumor, lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonal carcinoma, teratoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenomatoid tumor, lipoma); Liver: liver cancer (hepatocellular carcinoma), bile duct cancer, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; Biliary tract: gallbladder cancer, ampullary cancer, bile duct cancer; Bone: osteosarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cell sarcoma), multiple myeloma, malignant giant cell chordoma, enchondroma (osteochondral exostosis), benign enchondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma, and giant cell tumor; Nervous system: skull (osteomas, hemangiomas, granulomas, xanthomas, osteitis deformans), meninges (meningiomas, meningeal sarcomas, gliomas), brain (astrocytomas, medulloblastomas, gliomas, ependymomas, germinal cell tumors (pinealomas), glioblastoma multiforme, oligodendrogliomas, schwannomas, retinoblastomas, congenital tumors, spinal neurofibromas, meningiomas, gliomas, sarcomas); Gynecology: uterus (endometrial cancer (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosa-thecal cell tumor, Sertoli-Leydig cell tumor, germ cell tumor, malignant teratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma)), fallopian tube (cancer); Hematology: Blood (myeloid leukemia (acute and chronic), acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative disorders, multiple myeloma, myelodysplastic syndrome), Hodgkin's disease, non-Hodgkin's lymphoma (malignant lymphoma); Skin: malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, nevus dysplasia, lipoma, hemangioma, dermatofibroma, keloid, psoriasis; Adrenal gland: neuroblastoma.

9. The use according to claim 8, characterized in that The cancer is pancreatic cancer, lung cancer, colorectal cancer, cholangiocarcinoma, multiple myeloma, melanoma, uterine cancer, endometrial cancer, thyroid cancer, acute myeloid leukemia, bladder cancer, urothelial carcinoma, gastric cancer, cervical cancer, head and neck squamous cell carcinoma, diffuse large B-cell lymphoma, esophageal cancer, chronic lymphocytic leukemia, hepatocellular carcinoma, breast cancer, ovarian cancer, prostate cancer, glioblastoma, kidney cancer or sarcoma.

10. Use of a compound according to at least one of claims 1 to 5, a stereoisomer, enantiomer, diastereomer, deuterate, hydrate, solvate, metabolite, prodrug, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 6 in the preparation of a medicament for treating or preventing neurofibromatosis type 1 (NF1), Noonan syndrome (NS), Noonan syndrome with multiple lentigines (NSML), capillary malformation-arteriovenous malformation syndrome (CM-AVM), Costello syndrome (CS), cardiofacial-cutaneous syndrome (CFC), Leggs syndrome, and hereditary gingival fibromatosis.

11. Use of a compound according to at least one of claims 1 to 5, a stereoisomer, enantiomer, diastereomer, deuterate, hydrate, solvate, metabolite, prodrug, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 6, in the preparation of a medicament for treating and / or preventing a KRAS-mediated disease or condition, wherein the KRAS is preferably a mutant KRAS, and the mutant KRAS is preferably one or more of KRAS G12C, KRAS G12D, KRAS G13D, and KRAS G12V.