Substituted bicyclic heteroaryl compounds as kras g12d inhibitors

Abstract

The present invention provides substituted bicyclic heteroaryl compounds as KRAS G12D inhibitors. The present invention also provides pharmaceutical compositions comprising the compounds, and their use in the treatment of cancer.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field, specifically relating to substituted bicyclic heteroaryl compounds that can be used as KRAS G12D inhibitors. Background Technology

[0002] The human RAS gene family comprises three classes of RAS genes: KRAS, NRAS, and HRAS, encoding four different RAS proteins (KRAS-4A, KRAS-4B, NRAS, and HRAS). RAS proteins belong to the GTPase protein family. They are inactive when bound to GDP, but become active when bound to GTP, leading to the activation of downstream signaling pathways such as RAF-MAPK and PI3K-Akt, resulting in cell anti-apoptosis and proliferation (Cell, 2017; 170(1):17-33; Cell, 2020; 183(4):850-859; NatRev Drug Discov, 2020; 19(8):533-552). Activating mutations in RAS genes are the most common oncogene drivers in human cancers, with KRAS being the most frequently activating oncogene. For example, the mutation rate of KRAS is 86-96% in pancreatic cancer, 40-54% in colorectal cancer, and 27-39% in lung cancer (PNAS, 2019; 116(32): 15823-15829; Pathol Res Pract, 2009; 205, 858–862; Nature, 2012; 491, 399–405; Nature, 2014; 511, 543–550).

[0003] Oncogenic driver mutations can occur at multiple sites in the KRAS gene. The most common mutations occur at the G12 site, including G12C, G12D, and G12V. These mutations can reduce the GTPase activity of the KRAS protein, thereby causing the KRAS protein to remain in an active state for a long time, leading to malignant transformation of cells and cancer (Cell, 2017; 170(1):17-33; Nat RevDrug Discov, 2020; 19(8):533-552). The types of KRAS mutations that frequently occur vary across different cancer types. For example, approximately 13% of lung cancer patients have the G12C mutation (N Engl Med J, 2021; 384(25):2382-2393); while in pancreatic cancer, 33.8% of patients have the G12D mutation, but only 1.7% have the G12C mutation; in colorectal cancer, approximately 10-12% of patients have the G12D mutation, while the incidence of the G12C mutation is less than 3% (Nat Rev Cancer 2018; 18(12):767-777).

[0004] Unlike ATP-dependent protein kinases (where the affinity of proteins for ATP is at the micromolar level), KRAS proteins have an affinity for GDP / GTP at the picomolar level. Compounds struggle to effectively compete with GDP / GTP to inhibit the KRAS signaling pathway, severely hindering the development of KRAS inhibitors (Cell, 2017; 170(1):17-33; Cell, 2020; 183(4):850-859; Nat Rev Drug Discov, 2020; 19(8):533-552). In recent years, allosteric binding cavities that can effectively bind to small molecules without competing with GDP / GTP have been discovered on the KRAS protein. These findings have greatly facilitated the development of targeted drugs targeting KRAS mutation-driven tumors. Currently, MRTX-849 (Adagrasib) and AMG510 (Sotorasib), which target KRAS G12C, have demonstrated excellent efficacy in clinical studies (N Engl J Med. 2020; 383(13):1207-1217; Cancer Discov. 2020; 10(1):54-71), and AMG510 was successfully approved by the FDA in May 2021. Compared with the development of KRAS G12C drugs, the development of drugs targeting KRAS G12D mutations is still in its early stages; however, as shown above, there is a very high KRAS G12D mutation rate in many tumors, making the development of KRAS G12D inhibitors extremely important.

[0005] Mirati, a US company, reported intravenously administered KRAS G12D small molecule MRTX1133, but it has not yet entered clinical trials, and its oral absorption is weak. The KRAS G12D targeted small molecule drug of the present invention has excellent activity and higher in vivo exposure, and is expected to achieve oral administration in order to address unmet clinical needs. Summary of the Invention

[0006] In one aspect, the present invention provides a compound, or a pharmaceutically acceptable salt, isotopic variant, tautomer, stereoisomer, prodrug, polymorph, hydrate, or solvate thereof, wherein said compound is selected from:

[0007]

[0008]

[0009]

[0010]

[0011]

[0012]

[0013]

[0014]

[0015] In another aspect, the present invention provides a pharmaceutical composition comprising the compound of the present invention and optionally a pharmaceutically acceptable excipient.

[0016] In another aspect, the present invention provides pharmaceutical compositions comprising the compounds of the present invention and pharmaceutically acceptable excipients, and further comprising other therapeutic agents.

[0017] In another aspect, the present invention provides the use of the compounds of the present invention in the preparation of medicaments for treating and / or preventing diseases mediated by the KRAS G12D mutant protein.

[0018] In another aspect, the present invention provides a method for treating and / or preventing KRAS G12D mutant protein-mediated diseases in subjects, comprising administering the compound or composition of the present invention to the subject.

[0019] In another aspect, the present invention provides compounds or compositions thereof for the treatment and / or prevention of diseases mediated by the KRAS G12D mutant protein.

[0020] In specific implementation schemes, the diseases treated by this invention include cancers selected from the following: acute myeloid leukemia, juvenile cancer, childhood adrenocortical carcinoma, AIDS-related cancers (e.g., lymphoma and Kaposi's sarcoma), anal cancer, appendiceal cancer, astrocytoma, atypical teratoma, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brainstem glioma, brain tumor, breast cancer, bronchial tumor, Burkitt lymphoma, carcinoid tumor, atypical teratoma, embryonic tumor, germ cell tumor, primary lymphoma, cervical cancer, childhood cancer, chordoma, cardiac tumor, chronic lymphocytic leukemia (CLL), and chronic myeloid leukemia. (CML), Chronic myeloproliferative disorders, Colon cancer, Colorectal cancer, Craniopharyngioma, Cutaneous T-cell lymphoma, Extrahepatic ductal carcinoma in situ (DCIS), Embryonic tumors, CNS cancers, Endometrial cancer, Ependymoma, Esophageal cancer, Olfactory neuroblastoma, Ewing's sarcoma, Extracranial germ cell tumors, Gonadal germ cell tumors, Ocular cancer, Fibrohistocytoma of bone, Gallbladder cancer, Gastric cancer, Gastrointestinal carcinoid tumors, Gastrointestinal stromal tumors (GIST), Germ cell tumors, Gestational trophoblastic tumors, Hairy cell leukemia, Head and neck cancer, Heart cancer, Liver cancer, Hodgkin's lymphoma, Hypopharyngeal cancer, Intraocular melanoma, Islet cell tumor, Pancreas Neuroendocrine tumors, kidney cancer, laryngeal cancer, lip and oral cavity cancer, liver cancer, lobular carcinoma in situ (LCIS), lung cancer, lymphoma, metastatic squamous neck cancer with occult primary lesions, midline carcinoma, oral cancer, multiple endocrine tumor syndrome, multiple myeloma / plasma cell carcinoma, mycosis fungoides, myelodyplasia syndrome, myelodyplasia / myeloproliferative tumor, multiple myeloma, Merkel cell carcinoma, malignant mesothelioma, malignant fibrous histiocytoma and osteosarcoma of the bone, nasal cavity and sinus cancer, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer (NSCLC), oral cancer, lip and oral cavity cancer, oropharyngeal cancer, ovarian cancer, pancreas Cancer, papilloma, paraganglioma, sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pleural pulmonary blastoma, primary central nervous system (CNS) lymphoma, prostate cancer, rectal cancer, transitional cell carcinoma, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, skin cancer, gastric cancer, small cell lung cancer, small bowel cancer, soft tissue sarcoma, cell lymphoma, testicular cancer, laryngeal cancer, thymoma and thymic carcinoma, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, trophoblastic tumors, rare childhood cancers, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer, or virus-induced cancer, preferably pancreatic cancer, colorectal cancer, or non-small cell lung cancer.

[0021] Other objects and advantages of the invention will become apparent to those skilled in the art from the following detailed embodiments, examples and claims.

[0022] definition

[0023] The term "KRAS G12D" refers to a mutant form of the mammalian KRAS protein, which contains an amino acid substitution of glycine for aspartic acid at position 12.

[0024] As used herein, the term "pharmaceutically acceptable salt" refers to carboxylates and amino acid addition salts of the compounds of the present invention that are suitable for contact with patient tissues within the limits of reliable medical judgment, without producing undue toxicity, irritation, allergic reactions, etc., and are effective for their intended use in proportion to a reasonable benefit / risk ratio, including (where possible) zwitterionic forms of the compounds of the present invention.

[0025] The term "subject" in the administration includes, but is not limited to: humans (i.e., men or women of any age group, e.g., pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., young adults, middle-aged adults, or older adults)) and / or non-human animals, such as mammals, e.g., primates (e.g., cynomolgus monkeys, rhesus monkeys), cattle, pigs, horses, sheep, goats, rodents, cats, and / or dogs. In some embodiments, the subject is a human. In some embodiments, the subject is a non-human animal. The terms "human," "patient," and "subject" are used interchangeably herein.

[0026] The terms “disease,” “disorder,” and “symptom” are used interchangeably in this article.

[0027] Generally, the "effective amount" of a compound refers to the amount sufficient to elicit a target biological response. As will be understood by those skilled in the art, the effective amount of the compounds of the present invention can vary depending on factors such as the biological target, the pharmacokinetics of the compound, the disease being treated, the administration method, and the age, health status, and symptoms of the subject. Effective amounts include therapeutic effective amounts and prophylactic effective amounts.

[0028] The term "combination" and related terms refer to the simultaneous or sequential administration of the compounds of the present invention and other therapeutic agents. For example, the compounds of the present invention may be administered simultaneously or sequentially with other therapeutic agents in separate unit dosage forms, or simultaneously with other therapeutic agents in a single unit dosage form. Detailed Implementation Plan

[0029] In this document, “compounds of the present invention” refers to the following compounds, their pharmaceutically acceptable salts, enantiomers, diastereomers, solvates, hydrates or isotopic variants, and mixtures thereof.

[0030] In one embodiment, the present invention relates to a compound, or a pharmaceutically acceptable salt, isotopic variant, tautomer, stereoisomer, prodrug, polymorph, hydrate, or solvate thereof, wherein said compound is selected from:

[0031]

[0032]

[0033]

[0034]

[0035]

[0036]

[0037]

[0038]

[0039] The compounds of this invention may include one or more asymmetric centers and therefore may exist in a variety of stereoisomeric forms, such as enantiomers and / or diastereomers. For example, the compounds of this invention may be individual enantiomers, diastereomers, or geometric isomers (e.g., cis and trans isomers), or may be in the form of mixtures of stereoisomers, including racemic mixtures and mixtures rich in one or more stereoisomers. The isomers can be separated from the mixture by methods known to those skilled in the art, including chiral high-performance liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers may be prepared by asymmetric synthesis.

[0040] The compounds of this invention may also exist as tautomers. A compound is not limited to any particular tautomer, but is intended to encompass all tautomer forms.

[0041] Those skilled in the art will understand that organic compounds can form complexes with solvents, react in the solvent, or precipitate or crystallize out of the solvent. These complexes are called "solvates." When the solvent is water, the complex is called a "hydrate." This invention covers all solvates of the compounds of this invention.

[0042] The term "solvent" refers to a compound or its salt that is bound to a solvent and formed typically by a solvent decomposition reaction. This physical association may include hydrogen bonding. Common solvents include water, methanol, ethanol, acetic acid, DMSO, THF, diethyl ether, etc. The compounds described herein can be prepared, for example, in crystalline form and can be solvated. Suitable solvates include pharmaceutically acceptable solvates and further include stoichiometric and non-stoichiometric solvates. In some cases, the solvate will be separable, for example, when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid. "Solvent" includes solvates in solution and separable solvates. Representative solvates include hydrates, ethanolates, and methanolates.

[0043] The term "hydrate" refers to a compound that is bound to water. Typically, it is determined by the ratio of the number of water molecules contained in the hydrate to the number of molecules of the compound in the hydrate. Therefore, a hydrate of a compound can be represented, for example, by the general formula R·xH₂O, where R is the compound and x is a number greater than 0. A given compound can form more than one type of hydrate, including, for example, monohydrates (x is 1), lower hydrates (x is a number greater than 0 and less than 1, e.g., hemihydrates (R·0.5H₂O)), and polyhydrates (x is a number greater than 1, e.g., dihydrates (R·2H₂O) and hexahydrates (R·6H₂O)).

[0044] The compounds of this invention can be in amorphous or crystalline forms (polymorphs). Furthermore, the compounds of this invention can exist in one or more crystalline forms. Therefore, this invention encompasses all amorphous or crystalline forms of the compounds of this invention within its scope. The term "polymorph" refers to the crystalline form of a compound (or its salts, hydrates, or solvates) with a particular crystal packing arrangement. All polymorphs have the same elemental composition. Different crystalline forms typically have different X-ray diffraction patterns, infrared spectra, melting points, densities, hardness, crystal shapes, photoelectric properties, stability, and solubility. Recrystallization solvents, crystallization rates, storage temperatures, and other factors can lead to the dominance of one crystalline form. Various polymorphs of the compounds can be prepared by crystallization under different conditions.

[0045] The present invention also includes isotopically labeled compounds (isotopic variants) that are equivalent to those described in formula (A), but in which one or more atoms are replaced by atoms with atomic masses or mass numbers different from those commonly found in nature. Examples of isotopes that can be introduced into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, respectively, for example... 2 H, 3 H, 13 C 11 C 14 C 15N、 18 O、 17 O、 31 P, 32 P, 35 S, 18 F and 36 Cl. Other isotopes of the present invention containing the aforementioned isotopes and / or other atoms, their prodrugs, and pharmaceutically acceptable salts of said compounds or said prodrugs are all within the scope of this invention. Certain isotope-labeled compounds of the present invention, for example, those incorporating radioactive isotopes (e.g.,...) 3 H and 14 Those in category C) can be used for drug and / or substrate tissue distribution determination. Tritium, i.e. 3 H and carbon-14, i.e. 14 Carbon isotopes are particularly preferred because they are easy to prepare and detect. Subsequently, they are replaced by heavier isotopes, such as deuterium, i.e., 2 H, because higher metabolic stability can provide therapeutic benefits, such as prolonged in vivo half-life or reduced dosage requirements, may be preferred in some cases. Isotopically labeled compounds of formula (A) of the present invention and their prodrugs can generally be prepared by using readily available isotopically labeled reagents instead of non-isotopically labeled reagents when performing the processes described below and / or the techniques disclosed in the examples and preparation examples.

[0046] Furthermore, prodrugs are also included in the context of this invention. As used herein, the term "prodrug" refers to a compound which is converted in vivo, for example, by hydrolysis in the blood, into its active form having a medical effect. Pharmaceutically acceptable prodrugs are described in T. Higuchi and V. Stella, Prodrugs as Novel Delivery Systems, ACSSymposium Series, Vol. 14; Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987; and D. Fleisher, S. Ramon, and H. Barbra, "Improved oral drug delivery: solubility limitations overcome by the use of prodrugs," Advanced Drug Delivery Reviews (1996) 19(2) 115-130, each of which is incorporated herein by reference.

[0047] Pharmaceutical Compositions and Kits

[0048] In another aspect, the present invention provides pharmaceutical compositions comprising a compound of the present invention (also referred to as the "active component") and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition comprises an effective amount of the compound of the present invention. In some embodiments, the pharmaceutical composition comprises a therapeutically effective amount of the compound of the present invention. In some embodiments, the pharmaceutical composition comprises a preventatively effective amount of the compound of the present invention.

[0049] Pharmaceutically acceptable excipients used in this invention refer to non-toxic carriers, adjuvants, or mediators that do not impair the pharmacological activity of the compounds formulated together. Pharmaceutically acceptable carriers, adjuvants, or mediators that can be used in the compositions of this invention include (but are not limited to) ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (such as human serum albumin), buffering substances (such as phosphates), glycine, sorbic acid, potassium sorbate, mixtures of saturated vegetable fatty acid metaglycerides, water, salts or electrolytes (such as protamine sulfate), disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, silica gel, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and lanolin.

[0050] The present invention also includes a kit (e.g., a pharmaceutical package). The provided kit may include the compounds of the present invention, other therapeutic agents, and first and second containers (e.g., vials, ampoules, bottles, syringes, and / or dispersible packaging or other suitable containers) containing the compounds of the present invention and other therapeutic agents. In some embodiments, the provided kit may optionally include a third container containing pharmaceutical excipients for diluting or suspending the compounds of the present invention and / or other therapeutic agents. In some embodiments, the compounds of the present invention and other therapeutic agents provided in the first and second containers are combined to form a unit dosage form.

[0051] Dosage

[0052] The pharmaceutical compositions provided by this invention can be administered via a variety of routes, including but not limited to: oral administration, parenteral administration, inhalation administration, topical administration, rectal administration, nasal administration, oral administration, vaginal administration, administration via implantation, or other routes of administration. For example, parenteral administration as used herein includes subcutaneous administration, intradermal administration, intravenous administration, intramuscular administration, intra-articular administration, intra-arterial administration, intra-synovial administration, intrasternal administration, intramenstrual administration, intralesional administration, and intracranial injection or infusion techniques, preferably intravenous administration.

[0053] Typically, an effective amount of the compound described herein is administered. The actual amount of compound administered may be determined by the physician based on relevant circumstances, including the condition being treated, the chosen route of administration, the compound actually administered, the individual patient's age, weight and response, the severity of the patient's symptoms, etc.

[0054] When used to prevent the conditions described in this invention, the compounds provided herein are administered to subjects at risk of developing the conditions, typically based on a physician's advice and under physician supervision, at the dosage levels described above. Subjects at risk of developing a specific condition generally include subjects with a family history of the condition, or those identified through genetic testing or screening as particularly susceptible to developing the condition.

[0055] The pharmaceutical compositions provided herein can also be administered long-term (“long-term administration”). Long-term administration means administering the compound or a pharmaceutical composition thereof over a prolonged period of time, such as 3 months, 6 months, 1 year, 2 years, 3 years, 5 years, etc., or may be administered indefinitely, such as for the remainder of the subject's life. In some embodiments, long-term administration is intended to provide a constant level of said compound in the blood over a prolonged period of time, such as within a therapeutic window.

[0056] Various methods of administration can be used to further deliver the pharmaceutical composition of the present invention. For example, in some embodiments, the pharmaceutical composition can be administered by bolus injection, for instance, to increase the concentration of the compound in the blood to an effective level. The bolus dose depends on the target systemic level of the active component through the body; for example, an intramuscular or subcutaneous bolus dose results in a slow release of the active component, while a bolus dose delivered directly to a vein (e.g., via IV intravenous infusion) allows for a more rapid delivery, causing the concentration of the active component in the blood to rapidly increase to an effective level. In other embodiments, the pharmaceutical composition can be administered in the form of a continuous infusion, for example, via IV intravenous infusion, thereby providing a steady-state concentration of the active component in the subject's body. Furthermore, in other embodiments, a bolus dose of the pharmaceutical composition can be administered first, followed by a continuous infusion.

[0057] Oral compositions may be in the form of bulk liquid solutions, suspensions, or bulk powders. However, more commonly, the compositions are provided in unit dose form for the purpose of precise dosing. The term "unit dosage form" refers to a physically discrete unit suitable as a unit dose for human patients and other mammals, each unit containing a predetermined quantity of active substance and suitable pharmaceutical excipients suitable for producing the desired therapeutic effect. Typical unit dose forms include pre-filled, pre-measured ampoules or syringes for liquid compositions, or, in the case of solid compositions, pills, tablets, capsules, etc. In such compositions, the compound is typically a smaller component (about 0.1 to about 50% by weight, or preferably about 1 to about 40% by weight), with the remainder being various carriers or excipients useful for forming the desired dosage form, as well as processing aids.

[0058] For oral dosage, a typical regimen is one to five oral doses daily, particularly two to four oral doses, typically three oral doses. Using these dosage regimens, each dose provides approximately 0.01 to approximately 20 mg / kg of the compound of the invention, with preferred doses each providing approximately 0.1 to approximately 10 mg / kg, particularly approximately 1 to approximately 5 mg / kg.

[0059] To provide blood levels similar to or lower than those achieved with an injection dose, a transdermal dose is typically selected in an amount of about 0.01 to about 20% by weight, preferably about 0.1 to about 20% by weight, more preferably about 0.1 to about 10% by weight, and even more preferably about 0.5 to about 15% by weight.

[0060] From approximately 1 to approximately 120 hours, especially 24 to 96 hours, the injection dose level ranges from approximately 0.1 mg / kg / hour to at least 10 mg / kg / hour. To obtain adequate steady-state levels, a preload bolus of approximately 0.1 mg / kg to approximately 10 mg / kg or more may also be administered. For human patients weighing 40 to 80 kg, the maximum total dose should not exceed approximately 2 g / day.

[0061] Liquid forms suitable for oral administration may include suitable aqueous or non-aqueous carriers, as well as buffers, suspending and dispersing agents, colorants, flavoring agents, etc. Solid forms may include, for example, any of the following components, or compounds with similar properties: binders, such as microcrystalline cellulose, tragacanth gum, or gelatin; excipients, such as starch or lactose; disintegrants, such as alginic acid, Primogel, or corn starch; lubricants, such as magnesium stearate; gliding agents, such as colloidal silica; sweeteners, such as sucrose or saccharin; or flavoring agents, such as peppermint, methyl salicylate, or orange flavorings.

[0062] Injectable compositions are typically based on injectable sterile saline or phosphate-buffered saline, or other injectable excipients known in the art. As previously described, in such compositions, the active compound is typically a smaller component, often about 0.05 to 10% by weight, with the remainder being injectable excipients, etc.

[0063] Transdermal compositions are typically formulated as topical ointments or creams containing an active ingredient. When formulated as an ointment, the active ingredient is typically combined with a paraffin-based or water-miscible ointment base. Alternatively, the active ingredient may be formulated as a cream with, for example, an oil-in-water emulsion base. Such transdermal formulations are well known in the art and generally include other components to enhance stable skin penetration of the active ingredient or formulation. All such known transdermal formulations and components are included within the scope of this invention.

[0064] The compounds of this invention can also be administered via transdermal devices. Therefore, transdermal drug delivery can be achieved using reservoirs or porous membrane types, or patches with various solid matrices.

[0065] The above-described components for oral, injectable, or topical administration are merely representative. Other materials and processing techniques are described in Part 8 of Remington's Pharmaceutical Sciences, 17th edition, 1985, Mack Publishing Company, Easton, Pennsylvania, which is incorporated herein by reference.

[0066] The compounds of this invention can also be administered in a sustained-release form or from a sustained-release drug delivery system. Descriptions of representative sustained-release materials can be found at Remington's Pharmaceutical Sciences.

[0067] This invention also relates to pharmaceutically acceptable formulations of the compounds of this invention. In one embodiment, the formulation comprises water. In another embodiment, the formulation comprises a cyclodextrin derivative. The most common cyclodextrins are α-, β-, and γ-cyclodextrins, respectively, composed of 6, 7, and 8 α-1,4-linked glucose units, optionally including one or more substituents on the linked sugar moieties, including but not limited to: methylated, hydroxyalkylated, acylated, and sulfonyl ether substituted groups. In some embodiments, the cyclodextrin is a sulfonyl ether β-cyclodextrin, for example, sulfobutyl ether β-cyclodextrin, also known as Captisol. See, for example, US 5,376,645. In some embodiments, the formulation comprises hexapropyl-β-cyclodextrin (e.g., 10-50% in water).

[0068] Example

[0069] The reagents used in this invention are commercially available reagents that are purchased directly or synthesized using common methods known in the art.

[0070] The specific reaction routes or steps in the following examples are used in this invention, as detailed below:

[0071] Example 1

[0072] Preparation of key intermediates a1-a38

[0073] Synthesis of intermediate a1

[0074]

[0075] Step 1: Dissolve 15 g (64.1 mmol) of 2-amino-3-fluoro-4-bromobenzoic acid a1-1 in 100 mL of DMF, slowly add N-chlorosuccinimide (8.56 g, 64.1 mmol), heat to 80 °C and react for 16 hours, then stop the reaction. Pour the reaction solution into 500 mL of ice water, filter to obtain a filter cake, dry to give intermediate a1-2 (13.2 g, 49.2 mmol), yield: 77%. LC-MS: [MH] - =267.

[0076] Step 2: Add urea (35.3 g, 588 mmol) and intermediate a1-2 (10.5 g, 39.2 mmol) from the previous step to a 200 mL round-bottom flask, and heat to 200 °C and react for 12 hours. After cooling to 80 °C, add 100 mL of water to the system, reflux for 10 minutes, cool to room temperature, filter to obtain a filter cake, wash with water, and dry in an oven to obtain intermediate a1-3 (4.0 g, 13.7 mmol). Yield: 35%. LC-MS: [M+H] + =294.

[0077] Step 3: Dissolve intermediate a1-3 (4.0 g, 13.7 mmol) and N,N-diisopropylethylamine (5.3 g, 41.1 mmol) in 15 mL of phosphorus oxychloride, heat to 120 °C and react for 8 hours, then stop the reaction. Remove the solvent under reduced pressure, and separate by column chromatography to obtain intermediate a1 (1.9 g, 5.8 mmol). Yield: 42%. LC-MS: [M+H] + =331.

[0078] Synthesis of intermediates a2, a3, a16, a30

[0079]

[0080] Procedure: At room temperature, intermediate a1 (1.9 g, 5.8 mmol) and starting material 3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester a2-1 (1.85 g, 8.7 mmol) were dissolved in 20 mL of dichloromethane. N,N-diisopropylethylamine (2.3 g, 17.4 mmol) was added, and the mixture was reacted at room temperature for 8 hours. 60 mL of water was added to the system, and the mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by column chromatography to obtain a pale yellow solid a2 (1.2 g, 2.4 mmol). Yield: 41%. LC-MS: [M+H] + =507.

[0081] Following the synthetic route of intermediate a2, the following intermediate was synthesized.

[0082]

[0083]

[0084] Synthesis of intermediates a4-a6, a8-a13

[0085]

[0086] Step 1: Under ice bath conditions, 1-(methoxycarbonyl)cyclopropane-1-carboxylic acid a4-2 (3.0 g, 20.8 mmol) was dissolved in 60 mL of dichloromethane. Oxaloyl chloride (10.5 g, 83.3 mmol) was slowly added, followed by 5 drops of anhydrous DMF. The reaction was carried out at 0 °C for 20 minutes. The ice bath was removed, the temperature was raised to room temperature, and stirring was continued for 1 hour. The solvent was removed under reduced pressure. The mixture was then dissolved in 40 mL of dichloromethane, and N,N-diisopropylethylamine (5.4 g, 41.2 mmol) and tetrahydropyrrole a4-1 (1.78 g, 25.0 mmol) were added. The reaction was carried out at room temperature for 3 hours. The reaction was stopped, the solvent was removed under reduced pressure, and the mixture was separated by flash column chromatography to give intermediate a4-3 (2.6 g, 13.2 mmol). Yield: 64%. LC-MS: [M+H] + =198.

[0087] Step 2: At -78°C, intermediate a4-3 (2.6 g, 13.2 mmol) was dissolved in 30 mL of anhydrous tetrahydrofuran, and lithium aluminum hydride (26.4 mL, 1 M) was slowly added. The reaction was brought to room temperature for 2 hours, then stopped. 80 mL of ice water was slowly added to the reaction mixture, the organic solvent was removed by vacuum distillation, and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by flash column chromatography to obtain intermediate a4 (900 mg, 5.8 mmol). Yield: 44%. LC-MS: [M+H] + =156.

[0088] Following the synthetic route of intermediate a4, the following intermediate was synthesized.

[0089]

[0090]

[0091] Synthesis of intermediate a7

[0092]

[0093] Step 1: Under ice bath conditions, 25 g (113 mmol) of 6-bromo-2,3-difluorobenzaldehyde a7-1 was dissolved in 250 mL of methanol. Sodium borohydride (8.54 g, 226 mmol) was slowly added, and the mixture was stirred for 20 minutes. The temperature was then raised to room temperature, and the reaction was continued for 1 hour. The reaction mixture was then stopped. The solution was slowly poured into a saturated aqueous NH4Cl solution, extracted with dichloromethane, dried over anhydrous sodium sulfate, filtered, and concentrated to give a white solid a7-2 (23.9 g, 107 mmol). Yield: 95%.

[0094] Step 2: Under ice bath conditions, intermediate a7-2 (23.9 g, 107 mmol) and N,N-diisopropylethylamine (20.7 g, 161 mmol) were dissolved in 200 mL of anhydrous tetrahydrofuran. Methanesulfonic anhydride (20.5 g, 118 mmol) was slowly added, and the mixture was stirred for 20 minutes. The ice bath was then removed, and the reaction was continued at room temperature for 18 hours. The reaction mixture was then stopped. The solution was slowly poured into ice water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain an oily substance a7-3 (19 g, 63.1 mmol). Yield: 59%.

[0095] Step 3: Dissolve intermediate a7-3 (19 g, 63.1 mmol) from the previous step in 240 mL of a mixed solution of ethanol and water (v / v, 6 / 1), and add potassium cyanide (4.49 g, 69 mmol). React under reflux for 1 hour, then stop the reaction. Remove the organic solvent under reduced pressure, pour the reaction solution into a saturated sodium carbonate solution, stir for 20 minutes, extract with dichloromethane, concentrate, and separate by column chromatography to obtain intermediate a7-4 (10.4 g, 44.8 mmol). Yield: 71%. LC-MS: [M+H] + =233.

[0096] Step 4: Under ice bath conditions, intermediate a7-4 (10.4 g, 44.8 mmol) from the previous step was dissolved in 80 mL of DMF. Potassium tert-butoxide (5.3 g, 47.2 mmol) was slowly added, and the mixture was stirred for 20 minutes until the solution turned red. A pre-prepared DMF solution of ethyl isothiocyanate (6.2 g, 47.2 mmol, 5 mL) was then slowly added dropwise. The reaction mixture was stirred for another hour, then heated to 100 °C and reacted for 30 minutes. After cooling to room temperature, the reaction was quenched by slowly pouring ice water into the mixture. The mixture was filtered to obtain a filter cake, washed with n-hexane, and dried in a vacuum drying oven to obtain intermediate a7-5 (13.7 g, 40.0 mmol). Yield: 89%. LC-MS: [M+H] + =344.

[0097] Step 5: Dissolve intermediate a7-5 (6.7 g, 19.5 mmol) from the previous step in 30 mL of DMSO, and add 30 mL of sodium hydroxide aqueous solution (5 M). React under reflux for 4 hours, then stop the reaction. Cool to room temperature, and slowly add 100 mL of ice water to quench the reaction. Filter to obtain a filter cake, wash with water, and dry in a vacuum drying oven to obtain crude intermediate a7-6 (3.8 g). LC-MS: [M+H] + =272.

[0098] Step 6: Dissolve the crude product a7-6 (3.8 g) and DMAP (122 mg, 1 mmol) from the previous step in 30 mL of a THF / DMF mixed solution (v / v, 1 / 1), and add Boc anhydride (4.3 g, 19.5 mmol). React at room temperature for 12 hours, then stop the reaction. Add 80 mL of water to the system, extract with dichloromethane, dry to anhydrous sodium sulfate, filter, and concentrate to obtain the crude intermediate a7-7 (3.3 g).

[0099] Step 7: Under nitrogen protection, crude a7-7 (3.3 g) and starting material a7-8 (6.01 g, 26.7 mmol) were dissolved in 30 mL of 1,4-dioxane. Potassium acetate (2.62 g, 26.7 mmol) and Pd(DPEphos)Cl2 (643 mg, 0.9 mmol) were slowly added. The mixture was heated to 100 °C and reacted for 1 hour, then the reaction was stopped. After cooling to room temperature, the mixture was filtered, washed with saturated brine, extracted with dichloromethane, dried, concentrated, and separated by flash column chromatography to obtain intermediate a7 (2.5 g, 6.2 mmol). LC-MS: [M+H] + =405.

[0100] Synthesis of intermediates a14 and a17

[0101]

[0102] Procedure: Under nitrogen protection, intermediate a2 (50 g, 98.77 mmol) was dissolved in 750 mL of anhydrous DMF, and CsF (45 g, 29.63 mmol) was added. The mixture was heated to 60 °C and reacted for 8 h, after which the reaction was stopped. The reaction solution was added to 1 L of water, and the mixture was extracted three times with 750 mL of ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and filtered to obtain intermediate a14 in 90% yield. LC-MS: [M+H] + =489.

[0103] Following the synthetic route of intermediate a14, the following intermediate was synthesized.

[0104]

[0105] Synthesis of intermediates a15, a18, a33-a37

[0106]

[0107] Step 1: Dissolve 3-methoxy-2,2-dimethyl-3-oxomalacic acid a15-1 (1.0 g, 6.8 mmol) and a5-1 (850 mg, 6.8 mmol) in 20 mL of anhydrous dichloromethane. Add EDCI (1.56 g, 8.2 mmol), N,N-diisopropylethylamine (1.77 g, 13.6 mmol), and HOBt (1.1 g, 8.2 mmol). React at room temperature for 16 h, then stop the reaction. Add 60 mL of ice water to the reaction solution, extract with dichloromethane, and dry over anhydrous sodium sulfate. Separate the mixture by Flash column chromatography to obtain intermediate a15-2 (1.04 g, 4.6 mmol), yield 68%. LC-MS: [M+H] + =218.

[0108] Step 2: At -78°C, intermediate a15-2 (1.04 g, 4.6 mmol) was dissolved in 15 mL of anhydrous tetrahydrofuran, and lithium aluminum hydride (350 mg, 9.2 mmol) was slowly added. The mixture was heated to 0°C and reacted for 1 hour, then the reaction was stopped. 2 mL of 10% NaOH aqueous solution was slowly added to the reaction mixture, resulting in the precipitation of flocculent material. The precipitate was filtered, concentrated under reduced pressure, and separated by flash column chromatography to obtain intermediate a15 (700 mg, 4.0 mmol). Yield: 87%. LC-MS: [M+H] + =176.

[0109] Following the synthetic route of intermediate a15, the following intermediate was synthesized.

[0110]

[0111] Synthesis of intermediates a19-a23

[0112]

[0113] Step 1: In a 50 mL reaction flask, add (3S,4R)-4-fluoro-3-hydroxypyrrolidine a19-1 (300 mg, 2.86 mmol) and intermediate a19-2 (1.06 g, 3.14 mmol), and dissolve in 5 mL of anhydrous THF. After stirring for 5 minutes, add NaBH(OAc)3 (1.81 g, 8.58 mmol) and 5 drops of acetic acid to the reaction solution. React at room temperature for 10 hours. Stop the reaction, pour the reaction solution into 150 mL of ice water, extract with ethyl acetate, wash the organic phase with saturated brine, dry to anhydrous sodium sulfate, concentrate, and obtain 1.1 g of colorless oily crude intermediate a19-3. LC-MS: [M+H] + =428.

[0114] Step 2: Dissolve 1.1 g of crude intermediate a19-3 (1.1 g) from the previous step in 15 mL of anhydrous THF, add 3,4-dihydro-2H-pyran (390 mg) and p-toluenesulfonic acid (200 mg), stir at room temperature for 1 hour, and TLC shows that the reaction is complete. Pour the reaction solution into 80 mL of ice water, extract with ethyl acetate, dry to anhydrous sodium sulfate, concentrate, and obtain 1.5 g of crude intermediate a19-4. LC-MS: [M+H] + =514.

[0115] Step 3: Dissolve the intermediate a19-4 (1.5 g) from the previous step in 15 mL of tetrahydrofuran, and add tetrabutylammonium fluoride (1.5 g). React at room temperature for 3 hours, and monitor the reaction until complete by TLC. Dissolve the reaction solution in 50 mL of water, extract with ethyl acetate, dry to anhydrous sodium sulfate, concentrate, and separate the crude product by flash column chromatography (PE / EA = 1 / 1) to obtain a colorless oily substance a19 (200 mg, 0.73 mmol), yield: 35%, LC-MS: [M+H] + =274.

[0116] Following the synthetic route of intermediate a19, the following intermediate was synthesized.

[0117]

[0118]

[0119] Synthesis of intermediate a24

[0120]

[0121] Step 1: Add the starting material 3,3-difluorocyclobutane-1-amine a24-1 (1.0 g, 9.34 mmol) and intermediate a19-2 (3.16 g, 9.34 mmol) to a 100 mL reaction flask, and dissolve in 40 mL of anhydrous THF. After stirring for 5 minutes, add NaBH(OAc)3 (2.57 g, 12.34 mmol) and 10 drops of acetic acid to the reaction solution. React at room temperature for 10 hours. Stop the reaction, pour the reaction solution into 200 mL of ice water, extract with ethyl acetate, wash the organic phase with saturated brine, dry to anhydrous sodium sulfate, concentrate, and separate by flash column chromatography (PE / EA, 1 / 1) to obtain 2.62 g of colorless oily intermediate a24-2. LC-MS: [M+H] + =430.

[0122] Step 2: Add intermediate a24-2 (2.62 g, 5.91 mmol) and formaldehyde aqueous solution (0.26 mL) to a 100 mL reaction flask, and dissolve in 40 mL of tetrahydrofuran. After stirring for 5 minutes, add NaBH(OAc)3 (1.63 g, 7.68 mmol) and 10 drops of acetic acid to the reaction solution. React at room temperature for 2 hours. Stop the reaction, pour the reaction solution into 100 mL of ice water, extract with ethyl acetate, wash the organic phase with saturated brine, dry to anhydrous sodium sulfate, concentrate, and separate by flash column chromatography (PE / EA, 10 / 1) to obtain 2.0 g of colorless oily intermediate a24-3. LC-MS: [M+H] + =444.

[0123] Step 3: Dissolve intermediate a24-3 (2.0 g, 4.51 mmol) from the previous step in 20 mL of tetrahydrofuran, and add tetrabutylammonium fluoride (2.2 g, 9.0 mmol). React at room temperature for 12 hours, and monitor the reaction until complete using TLC. Dissolve the reaction solution in 50 mL of water, extract with ethyl acetate, dry to anhydrous sodium sulfate, concentrate, and separate the crude product by flash column chromatography (DCM / MeOH, 1 / 1) to obtain a colorless oily substance a24 (800 mg, 0.73 mmol), yield: 87%, LC-MS: [M+H] + =206.

[0124] Synthesis of intermediate a25

[0125]

[0126] Step 1: Dissolve starting materials a25-1 (1.0 g, 4.5 mmol) and a5-1 (680 mg, 5.4 mmol) in 50 mL of anhydrous dichloromethane. Add EDCI (1.29 g, 6.75 mmol), N,N-diisopropylethylamine (1.74 g, 13.5 mmol), and HOBt (910 mg, 6.75 mmol). React at room temperature for 4 h, then stop the reaction. Add 80 mL of ice water to the reaction solution, extract with dichloromethane, dry over anhydrous sodium sulfate, and concentrate to obtain crude intermediate a25-2 (1.21 g). LC-MS: [M+H] + =294.

[0127] Step 2: Under ice bath conditions, intermediate a25-2 (1.21 g, 4.13 mmol) was dissolved in 25 mL of anhydrous tetrahydrofuran, and lithium aluminum hydride (310 mg, 8.3 mmol) was slowly added. The reaction was heated to room temperature for 2 hours, then stopped. 2 mL of 10% NaOH aqueous solution was slowly added to the reaction mixture, precipitating flocculent material. This material was filtered, and the filtrate was concentrated under reduced pressure and separated by flash column chromatography to obtain intermediate a25 (180 mg, 0.76 mmol). Yield: 18%. LC-MS: [M+H] + =238.

[0128] Synthesis of intermediates a26 and a38

[0129]

[0130] Step 1: Add oxetane-3,3-dimethyldiethanol a26-1 (4.1 g, 33.9 mmol) and triethylamine (4.11 g, 40.63 mmol) to a 50 mL reaction flask, and dissolve in 40 mL of anhydrous dichloromethane. After stirring for 5 minutes, add tert-butyldiphenylchlorosilane (7.18 g, 33.86 mmol) to the reaction solution. React at room temperature for 2 hours, then stop the reaction. Pour the reaction solution into 200 mL of ice water, extract with dichloromethane, wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, and concentrate to obtain 12 g of crude intermediate a26-2.

[0131] Step 2: At -10℃, the crude intermediate a26-2 (12.0 g) from the previous step was dissolved in 40 mL of tetrahydrofuran, and pyridine trioxide (12.8 g, 81.0 mmol) was added. The reaction was carried out in an ice bath for 3 hours. The reaction was stopped, and the reaction solution was poured into 100 mL of ice water. The mixture was extracted with dichloromethane, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain intermediate a26-3 (8.0 g, 22.7 mmol). The yield of the two steps was 67%.

[0132] Step 3: Add intermediate a26-3 (8.0 g, 22.7 mmol) and intermediate a5-1 (3.4 g, 27.1 mmol) from the previous step to a 100 mL reaction flask, and dissolve in 40 mL of anhydrous THF. After stirring for 5 minutes, add NaBH(OAc)3 (7.18 g, 33.86 mmol) and 10 drops of acetic acid to the reaction solution. React at room temperature for 10 hours. Stop the reaction, pour the reaction solution into 200 mL of ice water, extract with ethyl acetate, wash the organic phase with saturated brine, dry to anhydrous sodium sulfate, and concentrate to obtain 5.0 g of crude intermediate a26-4. LC-MS: [M+H] + =428.

[0133] Step 4: Dissolve the crude product a26-4 (5.0 g, 11.69 mmol) from the previous step in 30 mL of tetrahydrofuran, and add tetrabutylammonium fluoride (4.58 g, 17.5 mmol). Heat to 40 °C and react for 12 hours, monitoring the reaction until complete via TLC. Dissolve the reaction solution in 150 mL of water, extract with ethyl acetate, dry to anhydrous sodium sulfate, concentrate, and separate the crude product by flash column chromatography (DCM / MeOH, 1 / 1) to obtain a colorless oily product a26 (1.9 g, 10.2 mmol), yield: 88%, LC-MS: [M+H] + =190.

[0134] Following the synthetic route of intermediate a26, the following intermediate was synthesized.

[0135]

[0136] Synthesis of intermediates a27-a28

[0137]

[0138] Step 1: At room temperature, intermediate a1 (3.0 g, 9.15 mmol) and starting material tert-butyl 4,7-diazaspiro[2.5]octane-4-carboxylate a27-1 (2.12 g, 10.1 mmol) were dissolved in 30 mL of tetrahydrofuran. N,N-diisopropylethylamine (2.3 g, 17.4 mmol) was added, and the mixture was reacted at room temperature for 8 hours. 100 mL of water was added to the system, and the mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by column chromatography to obtain a pale yellow solid a27-2 (3.74 g, 7.4 mmol). Yield: 81%. LC-MS: [M+H] + =505.

[0139] Step 2: Under nitrogen protection, intermediate a27-2 (3.74 g, 7.4 mmol) was dissolved in 75 mL of anhydrous DMF, and CsF (3.4 g, 22.2 mmol) was added. The mixture was heated to 60 °C and reacted for 8 h, after which the reaction was stopped. The reaction solution was added to 300 mL of water, and the mixture was extracted three times with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and filtered to obtain intermediate a27 in 90% yield. LC-MS: [M+H] + =489.

[0140] Following the synthetic route of intermediate a27, the following intermediate was synthesized.

[0141]

[0142] Synthesis of intermediate a29

[0143]

[0144] Step 1: Dissolve 2-fluoro-3-methyl-4-bromopyridine a29-1 (4.5 g, 23.9 mmol) in 25 mL of carbon tetrachloride. Slowly add N-bromosuccinimide (NBS) (6.35 g, 35.8 mmol) and azobisisobutyronitrile (AIBN) (390 mg, 2.3 mmol). React at room temperature for 3 hours, then stop the reaction. Slowly pour the reaction solution into 150 mL of ice water, extract with dichloromethane, dry to anhydrous sodium sulfate, filter, concentrate, and separate by column chromatography (PE / EtOAc, 5 / 1) to obtain an oily substance a29-2 (6.1 g, 22.9 mmol). Yield: 94%.

[0145] Step 2: Intermediate a29-2 (5.2 g, 3.7 mmol) and trimethylcyanosilane (TMSCN) (2.9 g, 5.6 mmol) were dissolved in 20 mL of acetonitrile. A solution of tetrahydrofuran (29 mL) containing TBAF (5.5 mmol) was added, and the reaction was carried out at room temperature for 16 hours. The reaction was then stopped. The solvent was removed under reduced pressure, and the mixture was separated by column chromatography (PE / EtOAc, 5 / 1) to obtain an oily substance a29-3 (3.4 g, 15.9 mmol). Yield: 81%.

[0146] Step 3: Under ice bath conditions, dissolve the intermediate a29-3 (3.4 g, 15.9 mmol) from the previous step in 20 mL of DMF. Slowly add NaH (2.9 g, 19.1 mmol), and stir for 30 minutes until the solution turns red. Slowly add dropwise a pre-prepared DMF solution of ethyl isothiocyanate (1.8 g, 15.9 mmol, 5 mL). Heat the reaction solution to 100 °C and react for 1 hour, then cool to room temperature. Slowly quench the reaction solution with ice water, extract with ethyl acetate, dry to anhydrous sodium sulfate, and separate the crude product by column chromatography (PE / EtOAc, 1 / 1) to obtain a pale yellow solid a29-4 (1.05 g, 3.2 mmol). Yield: 20%. LC-MS: [M+H] + =325.

[0147] Step 4: Dissolve the intermediate a29-4 (1.0 g, 3.1 mmol) from the previous step in 10 mL of DMSO, and add 10 mL of sodium hydroxide aqueous solution (5 M). React under reflux for 4 hours, then stop the reaction. Cool to room temperature, slowly add 100 mL of ice water to quench the reaction mixture, extract with ethyl acetate, dry to anhydrous sodium sulfate, and separate the crude product by column chromatography (PE / EtOAc, 1 / 1) to give a pale yellow solid a29-5 (450 mg, 1.8 mmol). Yield: 20%. LC-MS: [M+H] + =254.

[0148] Step 5: Dissolve intermediate a29-5 (450 mg, 1.8 mmol) and DMAP (5 mg, 0.04 mmol) from the previous step in 20 mL of a THF / DMF mixture (v / v, 1 / 1), and add Boc anhydride (465 mg, 2.16 mmol). React at room temperature for 12 hours, then stop the reaction. Add 50 mL of water to the system, extract with dichloromethane, dry to anhydrous sodium sulfate, filter, and concentrate to obtain crude intermediate a29-6 (760 mg).

[0149] Step 6: Under nitrogen protection, crude product a29-6 (760 mg) and starting material a7-8 (590 mg, 2.6 mmol) were dissolved in 10 mL of 1,4-dioxane. Potassium acetate (432 mg, 4.32 mmol) and Pd(DPEphos)Cl2 (46 mg, 0.065 mmol) were slowly added. The mixture was heated to 100 °C and reacted for 1 hour, then the reaction was stopped. After cooling to room temperature, the mixture was filtered, washed with saturated brine, extracted with dichloromethane, dried, concentrated, and separated by flash column chromatography to obtain intermediate a29 (320 mg, 0.82 mmol). LC-MS: [M+H] + =388.

[0150] Synthesis of intermediate a31

[0151]

[0152] Step 1: Under nitrogen protection, the starting material a31-1 (10.0 g, 40.5 mmol) was dissolved in 100 mL of anhydrous tetrahydrofuran, and LiAlH4 (2.3 g, 60.7 mmol) was slowly added. The reaction was carried out at room temperature for 2 hours. The reaction was stopped after LC-MS monitoring showed that the reaction was complete. 300 mL of ice water was added to the system, the organic solvent was removed by vacuum distillation, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by flash column chromatography to obtain a pale yellow oil a31-2 (5.9 g, 26.9 mmol), yield: 67%. LC-MS: [M+H]+=220.

[0153] Step 2: Under nitrogen protection and in an ice bath, dissolve intermediate a31-2 (5.9 g, 26.9 mmol) and imidazole (3.66 g, 53.8 mmol) in 50 mL of anhydrous dichloromethane, and slowly add tert-butyldiphenylchlorosilane (TBDPSCl) (7.4 g, 26.92 mmol). Stir the reaction mixture at room temperature for 30 minutes, monitor the reaction for completeness by LC-MS, and then stop the reaction. Add 100 mL of ice water to the reaction mixture, extract with dichloromethane, dry to anhydrous sodium sulfate, concentrate, and separate the crude product by column chromatography to obtain a pale yellow solid a31-3 (11.4 g, 24.9 mmol), yield: 93%. LC-MS: [M+H] + =458.

[0154] Step 3: Dissolve intermediate a31-3 (11.4 g, 24.93 mmol) from the previous step in a 1,4-dioxane solution of hydrogen chloride (4 M, 30 mL). Stir at room temperature for 1 hour, stop the reaction, filter, and dry the filter cake to obtain a31-4 hydrochloride (7.0 g, 17.8 mmol), yield: 71%. LC-MS: [M+H] + =358.

[0155] Step 4: Under nitrogen protection, the intermediate a31-4 (7.0 g, 17.8 mmol) from the previous step, the starting material cyclobutyl-1,1-dicarboxylic acid monoethyl ester a31-5 (4.6 g, 26.7 mmol), and DIEA (6.2 mL) were dissolved in 60 mL of DMF. HATU (10.2 g, 26.7 mmol) was added, and the reaction mixture was reacted at room temperature for 4 hours. The reaction was then stopped. 200 mL of ice water was added to the system, and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by column chromatography to obtain a brown solid a31-6 (7.78 g, 15.2 mmol), yield: 85%. LC-MS: [M+H] + =512.

[0156] Step 5: Dissolve the intermediate a31-6 (7.78 g, 15.2 mmol) from the previous step and tetrabutylammonium fluoride (TBAF) (7.96 g, 30.44 mmol) in 80 mL of tetrahydrofuran. Heat to 50 °C and react for 6 hours, then stop the reaction. Add 200 mL of water to the system, remove the organic solvent under reduced pressure, extract with ethyl acetate, dry to anhydrous sodium sulfate, concentrate, and separate the crude product by column chromatography to obtain a brown oily substance a31-7 (2.3 g, 8.42 mmol), yield: 55%. LC-MS: [M+H] + =274.

[0157] Step 6: Dissolve the intermediate a31-7 (2.3 g, 8.42 mmol), dihydropyran DHP (2.12 g, 25.3 mmol), and p-toluenesulfonic acid TsOH (145 mg, 0.84 mmol) in 20 mL of dichloromethane. Stir the reaction mixture at room temperature for 5 hours. Monitor the reaction progress by LC-MS until complete, then stop the reaction. Add 80 mL of ice water to the system, extract with dichloromethane, remove the solvent under reduced pressure, and separate the crude product by column chromatography to obtain the oily compound a31-8 (1.5 g, 4.2 mmol), yield: 50%. LC-MS: [M+H] + =358.

[0158] Step 7: Under nitrogen protection, the intermediate a31-8 (1.5 g, 4.2 mmol) from the previous step was dissolved in 20 mL of anhydrous tetrahydrofuran. LiAlH4 (319 mg, 8.40 mmol) was slowly added, and the mixture was stirred at room temperature for 2 hours. The reaction was monitored by LC-MS until complete. The reaction was quenched with 100 mL of ice water. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by column chromatography to give a brown solid a31 (760 mg, 2.5 mmol), yield: 60%. LC-MS: [M+H] + =302.

[0159] Synthesis of intermediate a32

[0160]

[0161] Step 1: In an ice bath, dissolve methyl 2-nitro-4-bromo-5-hydroxybenzoate a32-1 (25 g, 90.6 mmol) and triethylamine (27.5 g, 272 mmol) in 250 mL of dichloromethane, and slowly add acetyl chloride (10.66 g, 135.9 mmol) dropwise. After the addition is complete, continue the reaction for 3 hours, then stop the reaction. Add 500 mL of water to the system, extract with dichloromethane, and remove the solvent under reduced pressure to obtain the crude intermediate a32-2.

[0162] Step 2: Dissolve the crude product a32-2 and reduced iron powder (27.2 g, 487.3 mmol) in 310 mL of glacial acetic acid, heat to 40 °C and react for 2 hours, then stop the reaction. Filter the crude product through diatomaceous earth, and concentrate the filtrate under reduced pressure. Add saturated sodium bicarbonate aqueous solution to the filtrate to adjust the pH to approximately 8, extract with ethyl acetate, dry to anhydrous sodium sulfate, and concentrate to obtain crude intermediate a32-3 (16.7 g). LC-MS: [M+H] + =290.

[0163] Step 3: Dissolve the crude product a32-3 from the previous step and 1-chloromethyl-4-fluoro-1,4-diazabicyclo[2.2.2]octane di(tetrafluoroborate) salt (selectfluor, 30.8 g, 87.0 mmol) in 170 mL of acetonitrile. React at room temperature for 15 hours, then stop the reaction. Adjust the pH to approximately 8 by adding saturated sodium bicarbonate aqueous solution to the reaction solution, extract with ethyl acetate, dry to anhydrous sodium sulfate, concentrate, and separate the crude product by flash column chromatography to obtain compound a32-4 (1.6 g, 5.2 mmol). Three-step yield: 6%. LC-MS: [M+H] + =306.

[0164] Step 4: Dissolve the intermediate a32-4 (1.6 g, 5.2 mmol) and potassium carbonate (1.44 g, 10.5 mmol) in 16 mL of methanol. React at room temperature for 2 hours, then stop the reaction. Adjust the pH of the reaction solution to approximately 5 with 1 M dilute hydrochloric acid aqueous solution, extract with ethyl acetate, dry to anhydrous sodium sulfate, concentrate, and separate the crude product by flash column chromatography to obtain compound a32-5 (1.2 g, 4.54 mmol), with a yield of 87%. LC-MS: [M+H] + =264.

[0165] Step 5: In an ice bath, dissolve intermediate a32-5 (1.2 g, 4.54 mmol) and cesium carbonate (2.96 g, 9.1 mmol) from the previous step in 12 mL of DMF. Stir for 5 minutes, then slowly add iodomethane (710 mg, 5.0 mmol). After the addition is complete, continue the reaction for 2 hours, then stop the reaction. Add 50 mL of ice water to the reaction solution, extract with ethyl acetate, wash with saturated brine, concentrate under reduced pressure, and separate the crude product by column chromatography to obtain compound a32-6 (1.1 g, 3.96 mmol), in 87% yield. LC-MS: [M+H] + =278.

[0166] Steps 6 / 7: Dissolve the intermediate a32-6 (1.1 g, 3.96 mmol) from the previous step in 11 mL of methanol, add NaOH aqueous solution (791 mg, 19.8 mmol) dropwise, and react at room temperature for 6 hours. Then stop the reaction. Add potassium cyanide aqueous solution (963 mg, 11.88 mmol) to the reaction solution, heat to 50 °C, and react for 2 hours. During the reaction, adjust the pH to approximately 6.5 with 6 M dilute hydrochloric acid. Monitor the reaction completion by LC-MS, stop the reaction, and filter to obtain a gray solid a32-8 (1.1 g, 3.81 mmol). LC-MS: [M+H] + =291.

[0167] Step 8: Dissolve intermediate a32-8 (1.1 g, 3.81 mmol) and DIEA (984 mg, 7.62 mmol) in 11 mL of phosphorus oxychloride. Heat to 90 °C and react for 15 hours, then stop the reaction. Concentrate the reaction solution under reduced pressure. Slowly pour the crude product into 100 mL of ice water, precipitating a solid. Filter, wash the filter cake with water, and dry to obtain intermediate 11 (1.2 g, 3.68 mmol). Yield: 96.6%. LC-MS: [M+H] + =325.

[0168] Preparation of key intermediates b1-b10

[0169] Synthesis of intermediate b1

[0170]

[0171] Step 1: At room temperature, starting material b1-1 (10.00 g, 56.8 mmol), methoxyamine hydrochloride (6.73 g, 83.2 mmol), and pyridine (5.69 g, 68.10 mmol) were dissolved in 10 mL of anhydrous ethanol. The reaction solution was reacted at room temperature for 2 hours, then the reaction was stopped and concentrated under reduced pressure. The residue was dissolved in dichloromethane and washed with dilute hydrochloric acid (2N), saturated sodium bicarbonate aqueous solution, and saturated brine, respectively. The solution was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain crude colorless oily product b1-2 (11.30 g, 55.1 mmol). LC-MS: [M+H] + =206.

[0172] Step 2: Dissolve intermediate b1-2 (1.0 g, 4.9 mmol), palladium acetate (55 mg, 0.24 mmol), and NBS (0.87 g, 4.87 mmol) from the previous step in 10 mL of anhydrous acetic acid. Heat the reaction mixture to 80 °C and react for 1 hour. Stop the reaction, cool to room temperature, pour the reaction mixture into water, filter, and dry the filter cake to obtain a brown solid b1-3 (1.03 g, 3.6 mmol). LC-MS: [M+H] +=284.

[0173] Step 3: Dissolve intermediate b1-3 (12.5 g, 43.99 mmol) from the previous step in a mixed solution of 60 mL concentrated hydrochloric acid and 100 mL 1,4-dioxane. Heat the reaction mixture to reflux and stir for 1 hour. Stop the reaction and concentrate under reduced pressure. Dissolve the residue in ethyl acetate, wash with sodium hydroxide aqueous solution (1 N) and saturated brine, dry to anhydrous sodium sulfate, concentrate, and separate by flash column chromatography (PE / EA = 4 / 1) to give a yellow solid b1-4 (10.9 g, 42.7 mmol), yield: 97%. LC-MS: [M+H] + =255.

[0174] Step 4: Under nitrogen protection, intermediate b1-4 (7.90 g, 30.97 mmol) and 1-chloromethyl-4-fluoro-1,4-diazabicyclo[2.2.2]octane di(tetrafluoroborate) salt (Selectfluor, 16.46 g, 46.5 mmol) were dissolved in 80 mL of methanol, and 0.3 mL of concentrated sulfuric acid was slowly added dropwise. The reaction solution was heated to 50 °C and reacted for 5 hours. The reaction was then stopped and concentrated under reduced pressure. The residue was dissolved in ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by flash column chromatography (PE / EA = 10 / 1) to give red solid b1-5 (6.37 g, 23.35 mmol), yield: 75%. LC-MS: [M+H] + =273.

[0175] Step 5: Under nitrogen protection, intermediate b1-5 (32.0 g, 117.2 mmol) and pyridinium tribromide (41.22 g, 128.89 mmol) were dissolved in 300 mL of acetonitrile. The mixture was heated to 60 °C and reacted for 0.5 h. The reaction was then stopped, and the solvent was removed by vacuum distillation. The sample was washed with saturated brine, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by flash column chromatography (PE / EA = 10 / 1) to give a yellow solid b1-6 (36.0 g, 102.3 mmol), yield: 87%. LC-MS: [M+H] + =350.

[0176] Step 6: Under nitrogen protection, intermediate b1-6 (36.0 g, 102.3 mmol) and lithium bromide (19.5 g, 225 mmol) were dissolved in 100 mL DMF. The mixture was heated to 100 °C and reacted for 0.5 h, then the reaction was stopped. 300 mL of water was added to the system, and the mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated by flash column chromatography (PE / EA = 10 / 1) to give a pale yellow solid b1-7 (21.0 g, 77.5 mmol), yield: 75%. LC-MS: [M+H] + =271.

[0177] Step 7: Under nitrogen protection and in an ice bath, dissolve intermediate b1-7 (21.0 g, 77.5 mmol) and pyridine (18.38 g, 232.41 mmol) in 200 mL of dichloromethane. Slowly add trifluoromethanesulfonic anhydride (26.2 g, 92.96 mmol) dropwise to the reaction mixture, slowly raise the temperature to room temperature, and react for 1 hour. Stop the reaction and remove the solvent under reduced pressure. Wash the crude product with saturated brine, extract with dichloromethane, dry to anhydrous sodium sulfate, concentrate, and separate by flash column chromatography (PE / EA = 8 / 1) to give a yellow solid b1-8 (27.50 g, 68.2 mmol), yield: 88%. LC-MS: [M+H] + =403.

[0178] Step 8: Under nitrogen protection, intermediate b1-8 (1.0 g, 2.48 mmol), zinc cyanide (146 mg, 1.24 mmol), Pd2(dba)3 (114 mg, 0.12 mmol), and 1,1'-bis(diphenylphosphine)ferrocene (dppf, 137 mg, 0.25 mmol) were dissolved in 10 mL of anhydrous DMF. The reaction solution was heated to 70 °C and reacted for 3 hours, then cooled to room temperature. The crude product was poured into 50 mL of ice water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The crude product was separated by flash column chromatography (PE / EA = 5 / 1) to give a white solid b1-9 (270 mg, 0.96 mmol), yield: 38%. LC-MS: [M+H] + =208.

[0179] Step 9: Under nitrogen protection at -78°C, intermediate b1-9 (50 mg, 0.18 mmol) was dissolved in 2 mL of dichloromethane, and 0.44 mL of boron tribromide dichloromethane solution (2 M concentration) was slowly added dropwise. The system was slowly heated to 0°C and reacted for 16 hours, then quenched with 10 mL of methanol. The solvent was removed under reduced pressure, and the crude product was separated by flash column chromatography (PE / EA = 3 / 1) to give a white solid b1-10 (20 mg, 0.075 mmol), yield: 42%. LC-MS: [M+H] + =266.

[0180] Step 10: Under nitrogen protection, intermediate b1-10 (430 mg, 26.7 mmol), pinacol diboronate (823 mg, 3.24 mmol), potassium acetate (477 mg, 4.86 mmol), Pd2(dba)3 (74 mg, 0.081 mmol), and tricyclohexylphosphine (45 mg, 0.016 mmol) were dissolved in 8 mL of 1,4-dioxane. The mixture was heated to 105 °C and reacted for 10 hours, after which the reaction was stopped. The mixture was cooled to room temperature, filtered, and the system was poured into 30 mL of ice water. It was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The crude product was separated by flash column chromatography (PE / EA = 3 / 1) to give a pale yellow solid b1 (350 mg, 6.2 mmol), yield: 69%. LC-MS: [M+H]+ = 314.

[0181] Synthesis of intermediate b2

[0182]

[0183] Step 1: Dissolve 4-fluorophenylacetic acid b2-1 (50.0 g, 324.4 mmol) and cyclo(isopropyl)malonide b2-2 (51.4 g, 356.8 mmol) in 500 mL of acetonitrile, and add 4-dimethylaminopyridine (DMAP, 3.57 g, 29.2 mmol) and DIEA (88.0 g, 681.2 mmol). After stirring for 5 minutes, slowly add pentanoyl chloride (43.0 g, 356.8 mmol). Heat the reaction solution to 45 °C and stir for 3 hours, then cool to room temperature. Place the reaction solution in an ice bath, add 4N hydrochloric acid aqueous solution dropwise to adjust the pH to about 5, continue stirring for 1 hour, dilute with water, and adjust the pH of the reaction solution to about 2 again with 4N hydrochloric acid. A large amount of solid precipitates out. Filter, wash the filter cake with water, and dry to obtain white solid b2-3 (104 g, 371.1 mmol). The yield is quantitative. LC-MS:[M+H] + =281.

[0184] Step 2: The intermediate b2-3 (54.0 g, 192.7 mmol) from the previous step was slowly added to trifluoromethanesulfonic acid (228.5 g, 1.5 mol). The reaction mixture was stirred at room temperature for 2 hours, and the reaction was monitored for completeness by LC-MS. The reaction mixture was slowly poured into 500 mL of ice water, and a solid precipitated out. The solid was filtered, the filter cake was washed with water, and dried to obtain a brown solid b2-4 (66.0 g, 295.96 mmol). Yield: 93%, LC-MS: [M+H] + =223.

[0185] Step 3: Dissolve intermediate b2-4 (66.0 g, 295.96 mmol) from the previous step in a mixed solution of acetonitrile and water (v / 1, 1 / 1). Heat to 80 °C and react for 13 hours, then stop the reaction. Remove the solvent under reduced pressure, wash with saturated sodium bicarbonate solution, extract with ethyl acetate, dry to anhydrous sodium sulfate, and concentrate to give a pale yellow compound b2-5 (51.8 g, 291.01 mmol), yield: 97%. LC-MS: [M+H] + =179.

[0186] Step 4: Under nitrogen protection, crude product b2-5 (20.0 g, 112.3 mmol), (2-bromoethynyl)triisopropylsilane (30.8 g, 112.3 mmol), potassium acetate (22.0 g, 224.5 mmol), and dichlorobis(4-methylisopropylphenyl)ruthenium(II) (2.06 g, 3.4 mmol) were dissolved in 200 mL of 1,4-dioxane. The mixture was heated to 100 °C and reacted for 4 hours. The reaction was monitored by LC-MS until complete, and the mixture was filtered. The solvent was removed under reduced pressure, 100 mL of water was added to the system, and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated. The crude product was separated by flash column chromatography (PE / EA = 10 / 1) to give a pale yellow solid b2-6 (28.0 g, 78.2 mmol), yield: 69%. LC-MS: [M+H] + =359.

[0187] Step 5: Dissolve intermediate b2-6 (28 g, 78.2 mmol) and DIEA (20.19 g, 156.20 mmol) from the previous step in 300 mL of dichloromethane. Slowly add triisopropylchlorosilane (TIPSCl) (18.1 g, 93.7 mmol). After the addition is complete, stir the reaction mixture at room temperature for 1 hour. Monitor the reaction for completeness using LC-MS. Pour the reaction mixture into 500 mL of ice water, extract with dichloromethane, dry to anhydrous sodium sulfate, and concentrate to obtain crude crimson oily compound b2-7. LC-MS: [M+H] + =515.

[0188] Step 6: Under nitrogen protection, at -40°C, crude product b2-7 (58.4 g, 113.43 mmol) and DIEA (51.3 g, 397.0 mmol) were dissolved in 300 mL of dichloromethane. Trifluoromethanesulfonic anhydride (54.4 g, 192.8 mmol) was slowly added dropwise over 3 hours. The mixture was stirred for another 0.5 hours, and then the reaction was stopped. The reaction solution was poured into 500 mL of ice water, extracted with dichloromethane, dried over anhydrous sodium sulfate, and concentrated. The crude product was separated by flash column chromatography (PE / EA = 10 / 1) to give a pale red oily compound b2-8 (57.7 g, 89.2 mmol), yield: 78%. LC-MS: [M+H] + =647.

[0189] Step 7: Under nitrogen protection, intermediate b2-8 (61.6 g, 95.2 mmol), triethylamine (38.5 g, 380.9 mmol), and pinacolborane (48.7 g, 380.9 mmol) were dissolved in 600 mL of acetonitrile. After stirring for 5 minutes, the catalyst Pd(dppf)Cl2 (4.2 g, 5.7 mmol) was added. The reaction mixture was heated to 80 °C and stirred for 4 hours, then cooled to room temperature. The mixture was slowly quenched with MeOH while maintaining the temperature below 25 °C, resulting in the precipitation of a solid. The solid was filtered, the filter cake was washed with MeOH, and dried to give a white solid compound b2 (45.9 g, 73.4 mmol), yield: 77%. LC-MS: [M+H] + =625.

[0190] Synthesis of intermediate b3

[0191]

[0192] Step 1: Dissolve starting material b3-1 (6.0 g, 15.5 mmol) and AIBN (1.27 g, 7.75 mmol) in 60 mL of carbon tetrachloride, and slowly add N-bromosuccinimide (NBS) (13.8 g, 77.5 mmol). Heat to 60 °C and react for 6 hours, then stop the reaction. Remove the solvent under reduced pressure, and separate the residue by flash column chromatography (petroleum ether / ethyl acetate = 10 / 1) to give a pale yellow solid b3-2 (7.6 g, 14.0 mmol), yield: 90%. LC-MS: [M+H] + =543.

[0193] Step 2: Dissolve intermediate b3-2 (7.6 g, 14 mmol) and DIEA (1.8 g, 14.0 mmol) from the previous step in 80 mL of acetonitrile. Slowly add diethyl phosphite (1.95 g, 14.0 mmol). React at room temperature for 1 hour, then stop the reaction. Pour the reaction solution into 100 mL of ice water, extract with ethyl acetate, dry to anhydrous sodium sulfate, concentrate, and separate the crude product by flash column chromatography (petroleum ether / ethyl acetate = 10 / 1) to give a pale yellow solid b3-3 (4.6 g, 10.0 mmol), yield: 72%. LC-MS: [M+H] + =465.

[0194] Step 3: Dissolve intermediate b3-3 (4.6 g, 10 mmol), potassium carbonate (2.8 g, 20.0 mmol), and trimethylsilylcyanoTMSCN (1.5 g, 15.0 mmol) in 50 mL of acetonitrile. React at room temperature for 4 hours, then stop the reaction. Pour the reaction solution into 300 mL of ice water, extract with ethyl acetate, dry to anhydrous sodium sulfate, concentrate, and separate the crude product by flash column chromatography (petroleum ether / ethyl acetate = 3 / 1) to give a pale yellow solid b3-4 (3.6 g, 8.76 mmol), yield: 87%. LC-MS: [M+H] + =412.

[0195] Step 4: Under nitrogen protection in an ice bath, dissolve intermediate b3-4 (3.6 g, 8.76 mmol) from the previous step in 40 mL of anhydrous tetrahydrofuran. Add NaH (700 mg, 17.5 mmol), stir for 10 minutes, and then add 1,2-dibromoethane (2.0 g, 10.5 mmol) dropwise. After the addition is complete, heat to room temperature and react for 10 hours, then stop the reaction. Pour the reaction solution into 100 mL of ice water, extract with ethyl acetate, dry to anhydrous sodium sulfate, concentrate, and separate the crude product by flash column chromatography (petroleum ether / ethyl acetate = 3 / 1) to give a pale yellow solid b3 (1.2 g, 3.56 mmol), yield: 40%. LC-MS: [M+H] + =338.

[0196] Synthesis of intermediates b4-b6

[0197]

[0198] Step 1: Dissolve the starting material 4-bromo-7-fluoro-1H-indole (3.5 g, 16.4 mmol) in 35 mL of DMF, add N-iodosuccinimide (NIS) (3.69 g, 16.4 mmol), heat to 60 °C and react for 1 hour, then stop the reaction. Pour the reaction solution into 150 mL of ice water, extract with ethyl acetate, wash with saturated brine, dry to anhydrous sodium sulfate, concentrate, and separate the crude product by flash column chromatography (petroleum ether / ethyl acetate = 3 / 1) to give a pale yellow solid b4-2 (2.0 g, 5.9 mmol), yield: 36%. LC-MS: [M+H] + =340.

[0199] Step 2: Under nitrogen protection and in an ice bath, dissolve the intermediate 3-iodo-4-bromo-7-fluoro-1H-indole b4-2 (2.0 g, 5.9 mmol) from the previous step in 20 mL of anhydrous tetrahydrofuran. Add NaH (235 mg, 8.9 mmol) and iodomethane (92 mg, 6.5 mmol). Heat to room temperature and react for 2 hours, then stop the reaction. Add 80 mL of saturated ammonium chloride aqueous solution to the reaction mixture, extract with ethyl acetate, dry to anhydrous sodium sulfate, concentrate, and separate the crude product by flash column chromatography (petroleum ether / ethyl acetate = 3 / 1) to give a pale yellow solid b4-3 (1.1 g, 3.12 mmol), yield: 53%. LC-MS: [M+H] + =354.

[0200] Step 3: Under nitrogen protection, intermediate b4-3 (1.1 g, 3.12 mmol) and CuCN (1.1 g, 12.4 mmol) from the previous step were dissolved in 20 mL of DMSO. The mixture was heated to 100 °C and reacted for 3 hours, then cooled to room temperature. The reaction solution was poured into 150 mL of ice water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by flash column chromatography (petroleum ether / ethyl acetate = 2 / 1) to give a pale yellow solid b4-4 (0.55 g, 2.2 mmol), yield: 70%. LC-MS: [M+H] + =253.

[0201] Step 4: Under nitrogen protection, intermediate b4-4 (350 mg, 1.4 mmol), pinacol diboronate (706 mg, 2.8 mmol), and potassium acetate (408 mg, 4.2 mmol) from the previous step were dissolved in 10 mL of 1,4-dioxane. Catalyst Pd(dppf)Cl2 (100 mg, 0.14 mmol) was added, and the mixture was heated to 100 °C and reacted for 8 hours. The mixture was then cooled to room temperature. The reaction solution was poured into 50 mL of ice water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by flash column chromatography (petroleum ether / ethyl acetate = 2 / 1) to give a pale yellow solid b4 (0.27 g, 0.9 mmol), yield: 65%. LC-MS: [M+H] + =301.

[0202] Following the synthetic route of intermediate b4, the following intermediate was synthesized.

[0203]

[0204]

[0205] Synthesis of intermediate b7

[0206]

[0207] Step 1: Dissolve starting material b7-1 (1.0 g, 5.9 mmol) in 10 mL of DMF, add N-iodosuccinimide (NIS) (1.99 g, 8.85 mmol), heat to 70 °C and react for 1 hour, then stop the reaction. Pour the reaction solution into 50 mL of ice water, extract with ethyl acetate, wash with saturated brine, dry to anhydrous sodium sulfate, concentrate, and separate the crude product by flash column chromatography (petroleum ether / ethyl acetate = 9 / 1) to give a yellow solid b7-2 (800 mg, 2.70 mmol), yield: 46%. LC-MS: [M+H] + =296.

[0208] Step 2: Under nitrogen protection and in an ice bath, dissolve intermediate b7-2 (800 mg, 2.70 mmol), anhydrous methanol (430 mg, 13.6 mmol), and triphenylphosphine (1.07 g, 4.1 mmol) in 20 mL of DMF. Add diethyl azodicarbonate (710 mg, 4.1 mmol) to the reaction solution, heat to room temperature, and react for 12 hours. Then stop the reaction. Add 80 mL of ice water to the reaction solution, extract with ethyl acetate, wash with saturated brine, dry over anhydrous sodium sulfate, concentrate, and separate the crude product by flash column chromatography (petroleum ether / ethyl acetate = 5 / 1) to obtain a yellow solid b7-3 (530 mg, 1.72 mmol), yield: 64%. LC-MS: [M+H] + =310.

[0209] Step 3: Under nitrogen protection, intermediate b7-3 (530 mg, 1.71 mmol) and CuCN (610 mg, 5.84 mmol) from the previous step were dissolved in 20 mL of DMSO. The mixture was heated to 100 °C and reacted for 6 hours, then cooled to room temperature. The reaction solution was poured into 80 mL of ice water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by flash column chromatography (petroleum ether / ethyl acetate = 3 / 1) to give a yellow solid b7 (85 mg, 0.41 mmol), yield: 24%. LC-MS: [M+H] + =209.

[0210] Synthesis of intermediate b8

[0211]

[0212] Step 1: Under nitrogen protection, compound b8-1 (300 mg, 1.23 mmol) and ethyl isothiocyanate (240 mg, 1.84 mmol) were dissolved in 15 mL of 1,4-dioxane. The mixture was heated to 100 °C and reacted for 4 hours, after which the reaction was stopped. 50 mL of ice water was slowly added to the reaction solution, and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by flash column chromatography (PE / EA = 5 / 1) to give compound b8-2 (210 mg, 0.59 mmol), yield: 48%. LC-MS: [M+H] + =356.

[0213] Step 2: Dissolve intermediate b8-2 (210 mg, 0.59 mmol) from the previous step in 8 mL of DMSO, add 4 mL of sodium hydroxide aqueous solution (5 N), and react under reflux for 4 hours. Stop the reaction. Cool to room temperature, slowly add 50 mL of ice water to the reaction solution, extract with ethyl acetate, wash with saturated brine, dry over anhydrous sodium sulfate, concentrate, and separate the crude product by flash column chromatography (PE / EA = 3 / 1) to obtain compound b8-3 in quantitative yield. LC-MS: [M+H] + =283.

[0214] Step 3: Dissolve intermediate b8-3 (175 mg, 0.62 mmol), triethylamine (190 mg, 1.86 mmol), 4-dimethylaminopyridine (DMAP) (15 mg, 0.12 mmol), and di-tert-butyl dicarbonate (160 mg, 0.74 mmol) in 20 mL of dichloromethane. React at room temperature for 4 hours, then stop the reaction. Add 80 mL of water to the system, extract with dichloromethane, dry to anhydrous sodium sulfate, filter, and concentrate to obtain crude intermediate b8-4. LC-MS: [M+H] + =383.

[0215] Step 7: Under nitrogen protection, crude product b8-4 (100 mg, 0.26 mmol), potassium acetate (77 mg, 0.78 mmol), and pinacol diboronate (79 mg, 0.31 mmol) were dissolved in 7 mL of 1,4-dioxane. Palladium acetate catalyst (12 mg, 0.052 mmol) was added, and the mixture was heated to 100 °C and reacted for 3 hours. The reaction was then stopped. After cooling to room temperature, the mixture was filtered, and the solvent was removed by vacuum distillation. The crude product was separated by flash column chromatography (PE / EA = 5 / 1) to obtain intermediate b8 (60 mg, 0.14 mmol), yield: 54%. LC-MS: [M+H] + =431.

[0216] Synthesis of intermediate b9

[0217]

[0218] Step 1: Dissolve 2-amino-3-cyano-4-bromoindole b9-1 (3.8 g, 16.10 mmol) and 4-dimethylaminopyridine (DMAP) (196 mg, 1.61 mmol) in 50 mL of tetrahydrofuran. Slowly add di-tert-butyl dicarbonate (14.1 g, 64.4 mmol) and react at room temperature for 10 hours. Then stop the reaction. Add 200 mL of ice water to the reaction solution, extract with ethyl acetate, and concentrate to obtain crude b9-2. LC-MS: [M+H] + =436.

[0219] Step 2: Under nitrogen protection, the crude product b9-2 (600 mg, 1.38 mmol), potassium acetate (405 mg, 4.1 mmol), and pinacol diboronate (489 mg, 1.93 mmol) from the previous step were dissolved in 10 mL of 1,4-dioxane. Pd(dppf)Cl2 catalyst (102 mg, 0.14 mmol) was added, and the reaction was carried out at 100 °C for 2 hours. The reaction was then stopped. The mixture was filtered, and the solvent was removed under reduced pressure. The crude product was separated by flash column chromatography to obtain intermediate b9 (190 mg, 0.39 mmol). LCMS: [M+1] + =484.

[0220] Synthesis of intermediate b10

[0221]

[0222] Step 1: Under nitrogen protection, compound 4-bromo-benzofuran-2-carboxylic acid b10-1 (5.5 g, 23 mmol) and triethylamine (6.0 mL) were dissolved in 50 mL of tert-butanol. Diphenyl azidophosphate (DPPA) (9.4 g, 7.4 mL) was added, and the mixture was stirred for 10 minutes. The temperature was then raised to 95 °C and reacted for 3 hours. The reaction was stopped. The solvent was removed by vacuum distillation, and the crude product was dissolved in 50 mL of dichloromethane. Di-tert-butyl dicarbonate (5.0 g, 23 mmol) was added, and the mixture was reacted at room temperature for 2 hours. The reaction solution was poured into 100 mL of ice water, extracted with dichloromethane, and the solvent was removed by vacuum distillation. The crude product was separated by flash column chromatography to obtain compound b10-2 (4.0 g, 12.9 mmol), yield: 56%. LCMS: [M+1] + =312.

[0223] Step 2: Under nitrogen protection at -70℃, the intermediate b10-2 (1.0 g, 3.2 mmol) from the previous step was dissolved in 10 mL of anhydrous tetrahydrofuran. The starting material, chlorosulfonate isocyanate (0.42 mL), was added, and the mixture was stirred for 1 hour. Then, another 0.42 mL of chlorosulfonate isocyanate was added, and the reaction was continued for another hour. The temperature was then raised to room temperature. 10 mL of anhydrous DMF was added to the reaction solution, and the reaction was stopped after stirring for 1 hour. The reaction solution was poured into 100 mL of ice water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by flash column chromatography to obtain compound b10-3 (0.5 g, 1.5 mmol), with a yield of 46%. LCMS: [M+1] + =337.

[0224] Step 3: Under nitrogen protection, the intermediate b10-3 (0.5 g, 1.5 mmol), potassium acetate (440 mg, 4.5 mmol), and pinacol diboronate (460 mg, 1.8 mmol) from the previous step were dissolved in 10 mL of 1,4-dioxane. Pd(dppf)₂Cl₂ (11 mg, 0.15 mmol) was added, and the mixture was heated to 100 °C and reacted for 2 hours. The reaction was then stopped, and the mixture was filtered. 50 mL of ice water was added to the reaction solution, and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by flash column chromatography to obtain compound b10 (270 mg, 0.7 mmol), with a yield of 46%. LCMS: [M+1] + =385.

[0225] Preparation of key intermediates C1-C12

[0226] Synthesis of intermediates C1-C8

[0227]

[0228] Step 1: Under ice bath conditions, dissolve starting material c1-1 (20.0 g, 81.5 mmol) and TEA (16.5 g, 163.1 mmol) in 250 mL of dichloromethane. Slowly add methanesulfonic anhydride (15.6 g, 89.7 mmol) to the system. After the addition is complete, heat to room temperature and react for 2 hours, then stop the reaction. Add 300 mL of ice water to the system, extract with dichloromethane, dry over anhydrous sodium sulfate, and concentrate to obtain crude product c1-2. LC-MS: [M+H] + =324.

[0229] Step 2: Crude product c1-2 (25 g, 77.3 mmol) was dissolved in 300 mL of DMF. Sodium methanethiol (6.5 g, 92.8 mmol) was added at room temperature. The mixture was heated to 90 °C and stirred for 16 hours, then the reaction was stopped. The reaction solution was poured into 500 mL of ice water, extracted with ethyl acetate, washed with saturated brine, concentrated, and the crude product was separated by column chromatography to give compound c1-3 (10 g, 36.4 mmol), with a yield of 47%. LC-MS: [M+H] + =275.

[0230] Step 3: Under nitrogen protection at -78℃, intermediate C1-3 (16.5 g, 51.0 mmol) was dissolved in 200 mL of anhydrous tetrahydrofuran. LDA (12.8 g, 76.6 mmol) was slowly added dropwise. After the addition was complete, stirring was continued for 0.5 hours. 1-Bromo-3-chloropropane (40.2 g, 255.2 mmol) was slowly added dropwise to the system. After the addition was complete, the temperature was raised to room temperature and the reaction was continued for 1 hour. The reaction solution was then quenched with 100 mL of ice water. Extraction was performed with ethyl acetate, washing with saturated brine, drying over anhydrous sodium sulfate, and concentration to obtain crude product C1-4. LC-MS: [M+H] + =352.

[0231] Step 4: Dissolve the crude product C1-4 from the previous step in 50 mL of dichloromethane, add 150 mL of trifluoroacetic acid, stir at room temperature for 1 hour, and then stop the reaction. Remove the solvent under reduced pressure, adjust the mixture to weakly alkaline with saturated sodium bicarbonate aqueous solution, extract with ethyl acetate, dry to anhydrous sodium sulfate, concentrate, and purify the crude product by column chromatography to obtain compounds C1-5 (2.5 g) and C1-6 (1.3 g). LC-MS: [M+H] + =252.

[0232] Step 5: Dissolve intermediate C1-5 (2.5 g, 10.5 mmol), potassium iodide (0.17 g, 1.1 mmol), and potassium carbonate (6.9 g, 21.0 mmol) in 25 mL of methanol. React at room temperature for 16 hours, then stop the reaction. Add 100 mL of ice water to the system, extract with ethyl acetate, wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate to obtain crude product C1-7. LC-MS: [M+H] + =216.

[0233] Step 6: In an ice bath, dissolve crude c1-7 (2.1 g, 9.8 mmol) in 20 mL of tetrahydrofuran, add lithium aluminum hydride (750 mg, 19.5 mmol), and continue stirring for 1 hour. Stop the reaction. Quench the mixture with 10 mL of methanol, filter, concentrate under reduced pressure, add 50 mL of water, extract with dichloromethane, dry to anhydrous sodium sulfate, and concentrate to obtain crude c1 (for direct use in the next reaction). LC-MS: [M+H] + =188.

[0234] Step 7: Replace intermediate c1-5 with c1-6 to obtain intermediate c2.

[0235] Following the synthetic route of intermediate c1, the following intermediate was synthesized.

[0236]

[0237]

[0238] Synthesis of intermediate C9

[0239]

[0240] Step 1: Ice bath. Dissolve starting material C9-1 (15.0 g, 71.0 mmol) in 150 mL of anhydrous tetrahydrofuran (150 mL), and slowly add sodium borohydride (806 mg, 21.3 mmol). React in an ice bath for 3 hours, then stop the reaction. Remove the solvent under reduced pressure, add 50 mL of ice water to the mixture, extract with ethyl acetate, dry to anhydrous sodium sulfate, and concentrate. Separate the crude product by column chromatography to obtain intermediate C9-2 (12.0 g, 56.3 mmol). LC-MS: [M+H] + =214.

[0241] Step 2: Ice bath. Dissolve the intermediate C9-2 (12.0 g, 56.3 mmol) and triethylamine (17.3 g, 170.5 mmol) from the previous step in 120 mL of dichloromethane. After stirring for 5 minutes, add dropwise 20 mL of a dichloromethane solution of methanesulfonic anhydride (14.9 g, 85.2 mmol). After the addition is complete, continue stirring for 1 hour. Stop the reaction, add 200 mL of ice water to the system, extract with dichloromethane, concentrate under reduced pressure to obtain the crude product C9-3.

[0242] Step 3: Crude product C9-3 and potassium thioacetate (9.4 g, 82.4 mmol) were dissolved in 150 mL of DMF, and the mixture was heated to 60 °C and reacted for 15 hours. The reaction was then stopped. 300 mL of ice water was added to the system, and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was separated by column chromatography to obtain intermediate C9-4 (15 g, 55.4 mmol). Two-step yield: 10%. LC-MS: [M+H] + =272.

[0243] Step 4: Dissolve intermediate C9-4 (15 g, 55.4 mmol) from the previous step in 300 mL of anhydrous tetrahydrofuran, and add lithium aluminum hydride (5.2 g, 138 mmol). After stirring for 5 minutes, remove the ice bath and heat to 60 °C for 2 hours. Stop the reaction. Quench the reaction with dilute hydrochloric acid, adjust the pH to approximately 7, precipitate a solid, filter, and wash the filter cake with ethyl acetate to obtain intermediate C9. LC-MS: [M+H] + =174.

[0244] Synthesis of intermediate C10

[0245]

[0246] Procedure: Intermediate C9 (2.0 g, 11.5 mmol) was dissolved in 20 mL of DMF, and NaH (920 mg, 23 mmol) was added. The reaction was carried out at room temperature for 30 minutes. The temperature was lowered to -40 °C, and 5 mL of a DMF solution containing 3.4 g, 17.3 mmol of trifluoroiodomethane was added. The mixture was slowly heated to room temperature and stirred for 1 hour, at which point the reaction was stopped. 50 mL of ice water was added to the reaction mixture, and the mixture was extracted with ethyl acetate, washed with saturated brine, concentrated under reduced pressure, and separated by flash column chromatography to obtain intermediate C10 (1.3 g, 5.4 mmol), in 47% yield. LC-MS: [M+H] + =242.

[0247] Synthesis of intermediate C11

[0248]

[0249] Procedure: In an ice bath, intermediate C9 (2.2 g, 12.7 mmol) was dissolved in 30 mL of DMSO, and sodium hydride (1.02 g, 25.9 mmol) was added. After stirring for 30 minutes, bromocyclopropane (2.3 g, 19.0 mmol) was added, and the reaction was stopped at room temperature for 1 hour. 100 mL of ice water was added to the system, and the mixture was extracted with ethyl acetate, washed with saturated brine, and concentrated under reduced pressure to obtain intermediate C11. LC-MS: [M+H] + =214.

[0250] Synthesis of intermediate C12

[0251]

[0252] Procedure: In an ice bath, intermediate C9 (1.6 g, 9.2 mmol) was dissolved in 30 mL of DMSO, and sodium hydride (738 mg, 18.4 mmol) was added. After stirring for 30 minutes, bromocyclobutane (1.87 g, 13.9 mmol) was added, and the reaction was stopped at room temperature for 1 hour. 100 mL of ice water was added to the system, and the mixture was extracted with ethyl acetate, washed with saturated brine, and concentrated under reduced pressure to obtain intermediate C12. LC-MS: [M+H] + =228.

[0253] Preparation of key intermediates d1-d5

[0254] Synthesis of intermediate d1

[0255]

[0256] Step 1: Ice bath. In a 250 mL reaction flask, dissolve starting material d1-1 (5.0 g, 18.6 mmol) in 80 mL of pyridine. Slowly add 20 mL of oxaloyl chloride monoethyl ester dropwise to the system. After the addition is complete, stir at room temperature for 1 hour, then heat to 50 °C and react for 2 hours. Stop the reaction. Slowly pour the reaction solution into 300 mL of ice water, filter, and dry the filter cake to obtain intermediate d1-2 (5.2 g, 14.8 mmol), yield 80%. LC-MS: [M+H] + =350.

[0257] Step 2: Dissolve intermediate d1-2 (5.2 g, 14.8 mmol) from the previous step in 100 mL of anhydrous ethanol, add 4.0 g of ammonium acetate and 1 mL of acetic acid. Refrigerate under reflux for 1 hour, then cool to room temperature. Slowly add 1 M dilute hydrochloric acid to adjust the pH to approximately 5, causing a solid to precipitate. Filter the solid, dry the filter cake, and obtain crude product d1-3. LC-MS: [M+H] + =349.

[0258] Step 3: Dissolve the crude product d1-3 and DIEA (2.2 g, 17.2 mmol) from the previous step in 20 mL of phosphorus oxychloride, heat to 85 °C and react for 2 hours, then stop the reaction. Remove the solvent under reduced pressure, slowly pour the crude product into 100 mL of water, extract with ethyl acetate, dry to anhydrous sodium sulfate, concentrate, and separate by column chromatography to obtain compound d1-4 (2.5 g, 6.8 mmol), with a two-step yield of 46%. LC-MS: [M+H] + =367.

[0259] Step 4: Using an ice bath, dissolve intermediate d1-4 (2.5 g, 6.8 mmol) and DIEA (1.75 g, 13.6 mmol) from the previous step in 20 mL of anhydrous tetrahydrofuran. Slowly add 3.0 mL of a pre-prepared THF solution of a2-1 (1.44 g, 6.8 mmol) to the reaction solution. After the addition is complete, heat to room temperature and react for 40 minutes, then stop the reaction. Add 100 mL of water to the system, extract with ethyl acetate, dry over anhydrous sodium sulfate, and concentrate to give compound d1 (3.5 g, 6.4 mmol), yield 94%. LC-MS: [M+H] + =543.

[0260] Synthesis of intermediate d2

[0261]

[0262] Procedure: Under nitrogen protection, intermediates d1 (2.0 g, 3.7 mmol) and a7 (2.1 g, 1.02 mmol) were dissolved in 50 mL of toluene. Cesium carbonate (3.6 g, 11.1 mmol) and catalyst Pd(DPEPhos)Cl2 (600 mg, 1.1 mmol) were added sequentially. The reaction mixture was heated to 105 °C and reacted for 4 hours, then cooled to room temperature. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by flash column chromatography to give compound d2 (1.4 g, 1.85 mmol), yield: 50%. LC-MS: [M+H] + =755.

[0263] Synthesis of intermediate d3

[0264]

[0265] Step 1: In an ice bath, dissolve the starting material N-Boc-2-amino-2-methyl-1-propanol d3-1 (500 mg, 2.64 mmol) in 10 mL of dichloromethane. Slowly add Dess Martin oxidant (1.68 g, 3.96 mmol), heat to room temperature, and react for 2 hours. Then stop the reaction. Pour the reaction solution into 100 mL of water, extract with dichloromethane, and remove the solvent under reduced pressure. Separate the crude product by flash column chromatography (PE / EA = 4 / 1) to obtain an oily substance d3-2 (380 mg, 2.02 mmol), yield: 76%.

[0266] Step 2: Under nitrogen protection, the intermediate d3-2 (380 mg, 2.02 mmol) and the starting material (R)-3-fluoro-tetrahydropyrrole a5-1 (303 mg, 2.42 mmol) from the previous step were dissolved in 10 mL of anhydrous THF, and sodium triacetoxyborohydride (852 mg, 4.04 mmol) was slowly added. The reaction solution was reacted at room temperature for 10 hours, and then the reaction was stopped. 50 mL of ice water was added to the system, and the mixture was extracted with dichloromethane. The solvent was removed by vacuum distillation. The crude product was separated by flash column chromatography (PE / EA = 2: / 1) to give a white solid d3-3 (250 mg, 0.96 mmol), yield: 48%. LC-MS: [M+H] + =261.

[0267] Step 3: Under nitrogen protection, the intermediate d3-3 (250 mg, 0.96 mmol) from the previous step was dissolved in 8 mL of a mixed solution of trifluoroacetic acid and dichloromethane (v / v, 1 / 1). The reaction solution was reacted at room temperature for 1 hour. The reaction was then stopped, and saturated sodium bicarbonate solution was slowly added to adjust the pH to approximately 8. The mixture was extracted with ethyl acetate and concentrated under reduced pressure. The residue was separated by flash column chromatography to obtain a yellow oily substance d3 (200 mg). LC-MS: [M+H]+ =161.

[0268] Following the synthetic route of intermediate d3, the following intermediate was synthesized.

[0269]

[0270] Example 2:

[0271]

[0272] Step 1: Under nitrogen protection, intermediate a3 (2.4 g, 5.61 mmol) was dissolved in 50 mL of dioxane, and starting material P1-1 (1.34 g, 8.42 mmol) and N,N-diisopropylethylamine (DIEA) (1.45 g, 11.2 mmol) were added. The reaction mixture was stirred at 80 °C for 12 hours and then cooled to room temperature. 100 mL of water was added to the system, and the mixture was extracted with ethyl acetate, dried, filtered, and the solvent was removed under reduced pressure. The mixture was purified by column chromatography (petroleum ether:ethyl acetate = 3:1) to give compound P1-2 (2.7 g, 4.9 mmol). Yield: 87%, LC-MS: [M+H] + =552.

[0273] Step 2: Under nitrogen protection, compound P1-2 (300 mg, 0.54 mmol) and potassium phosphate (229 mg, 1.08 mmol) from the previous step were mixed in 6 mL of anhydrous toluene. Then, intermediate a7 (262 mg, 0.65 mmol), Xphos Pd G3 (93 mg, 0.11 mmol), and Xphos (53 mg, 0.11 mmol) were added sequentially. The reaction mixture was reacted at 100 °C for 1 hour under nitrogen protection. The reaction was stopped, cooled to room temperature, filtered, and the solvent was removed by vacuum distillation. The residue was separated by TLC to give compound P1-3 (120 mg, 0.15 mmol). Yield: 28%, LC-MS: [M+H] + =807.

[0274] Step 3: Under ice bath conditions, P1-3 (120 mg, 0.15 mmol) was dissolved in 4 mL of a mixed solution of trifluoroacetic acid and dichloromethane (v / v, 1 / 1). The reaction mixture was placed under nitrogen protection and the reaction was continued for 1 hour, after which the reaction was stopped. Saturated sodium bicarbonate solution was slowly added to the system and the pH was adjusted to approximately 8. The mixture was extracted with ethyl acetate, the solvent was removed under reduced pressure, and the residue was purified by preparative HPLC to obtain the target compound P1 (10.2 mg). LC-MS: [M+H] + =607.

[0275] 1H NMR (400MHz, DMSO-d6) δ9.07(s,1H),8.08(s,2H),7.41(dd,J=8.4,5.3Hz,1H),7.14(dd,J=9.5,8.4 Hz,1H),5.27(br,J=72Hz,1H),4.42(d,J=12.3Hz,2H),4.13(d,J=10.4Hz,1H),4.03(d,J=10.3Hz,1 H),3.71–3.52(m,4H),3.13–3.06(m,2H),3.01(s,1H),2.83(q,J=8.5Hz,1H),2.35–2.27(m,1H),2. 17–2.10(m,1H),2.08–1.98(m,2H),1.88–1.74(m,3H),1.65(d,J=6.6Hz,2H),1.56(d,J=7.5Hz,2H).

[0276] Following the synthetic route of compound P1, and using a similar skeletal structure, the following target molecule was synthesized.

[0277]

[0278]

[0279] Example 3:

[0280]

[0281] Step 1: Under nitrogen protection, intermediate a2 (500 mg, 0.99 mmol) was dissolved in 6 mL of anhydrous DMF. Then, starting material P1-1 (314 mg, 2.0 mmol) and cesium carbonate (966 mg, 2.96 mmol) were added. Under nitrogen protection, the reaction mixture was reacted at 140 °C for 2 hours. The reaction was then stopped and cooled to room temperature. 30 mL of water was added to the system, and the mixture was extracted with ethyl acetate. The solvent was removed by vacuum distillation. The residue was purified by TLC (petroleum ether:ethyl acetate = 1:4) to give a pale yellow solid H1-1 (110 mg, 0.18 mmol). Yield: 18%, LC-MS: [M+H] + =630.

[0282] Step 2: Under nitrogen protection, H1-1 (210 mg, 0.34 mmol) was dissolved in 5 mL of anhydrous toluene, followed by the sequential addition of intermediate a7 (189 mg, 0.47 mmol), Pd(DPEPhos)Cl2 (72 mg, 0.10 mmol), and anhydrous cesium carbonate (273 mg, 0.84 mmol). The reaction mixture was reacted at 105 °C for 6 hours under nitrogen protection. The reaction was then stopped, cooled to room temperature, and the solvent was removed under reduced pressure. The residue was separated by TLC (petroleum ether: ethyl acetate = 1:2) to give a yellow solid H1-2 (80 mg, 0.10 mmol). Yield: 28%, LC-MS: [M+H] + =841.

[0283] Step 3: Under ice bath conditions, H1-2 (80 mg, 0.10 mmol) was dissolved in 3 mL of a mixed solution of trifluoroacetic acid and dichloromethane (v / v, 1 / 1). Under nitrogen protection, the reaction mixture was allowed to react at room temperature for 0.5 hours. The reaction was then stopped, and a saturated sodium bicarbonate solution was slowly added to adjust the pH to approximately 8. The mixture was extracted with ethyl acetate and concentrated under reduced pressure. The residue was purified using preparative SFC (Xselect CSH C18 OBD) to obtain the target compounds H1a (4.0 mg) and H1b (4.1 mg).

[0284] H1a: 1 H NMR (300MHz, DMSO-d6) δ8.23(s,1H),8.11(s,1H),7.85(s,1H),7.26(dd,J=8. 4,5.3Hz,1H),7.15(dd,J=9.5,8.4Hz,1H),5.27(br,J=72Hz,1H),4.29(dd,J= 20.9,12.2Hz,3H),4.14–3.91(m,2H),3.67–3.39(m,5H),3.21–2.95(m,3H),2 .92–2.70(m,1H),2.14(d,J=6.8Hz,1H),2.09–1.93(m,2H),1.88–1.52(m,6H).

[0285] H1b: 1H NMR(400MHz,DMSO-d6)δ8.21(s,1H),8.09(s,2H),7.84(s,1H),7.26(dd,J=8.4,5.3Hz,1H),7 .18–7.11(m,1H),5.27(br,J=72Hz,1H),4.27(dd,J=19.6,12.0Hz,2H),4.08(d,J=10.3Hz,1H ),3.99(d,J=10.3Hz,1H),3.49–3.55(m,3H),3.08(d,J=9.5Hz,3H),3.02(d,J=11.5Hz,2H),2 .81(s,1H),2.14–2.12(m,1H),2.02(d,J=18.1Hz,2H),1.91–1.71(m,4H),1.66–1.54(m,4H).

[0286] Example 4:

[0287]

[0288] Step 1: In a 50 mL reaction flask, intermediate a5 (420 mg, 2.42 mmol) was dissolved in 10 mL of anhydrous THF, and potassium tert-butoxide (340 mg, 3.64 mmol) was added. The mixture was stirred at room temperature for 30 minutes to obtain solution S1. In another 50 mL reaction flask, intermediate a14 (1.0 g, 2.0 mmol) was dissolved in 10 mL of anhydrous THF. Under ice bath conditions, the prepared solution S1 was slowly added, and stirring was continued for 1 hour. The reaction was then stopped. The reaction solution was poured into 100 mL of ice water, extracted with ethyl acetate, and the solvent was removed by vacuum distillation. The residue was purified by flash column chromatography (petroleum ether: ethyl acetate = 1:1) to give a pale yellow solid H2-1 (916 mg, 1.42 mmol). Yield: 71%, LC-MS: [M+H] + =644.

[0289] Step 2: Under nitrogen protection, H2-1 (480 mg, 0.75 mmol) was dissolved in 10 mL of anhydrous toluene, followed by the sequential addition of intermediate a7 (414 mg, 1.02 mmol), Pd(DPEPhos)Cl2 (22 mg, 0.03 mmol), and cesium carbonate (438 mg, 1.35 mmol). The reaction mixture was reacted at 105 °C for 10 hours under nitrogen protection. The reaction was then stopped, cooled to room temperature, and the solvent was removed under reduced pressure. The residue was separated by flash column chromatography (petroleum ether: ethyl acetate = 1:2) to give a yellow solid H2-2 (450 mg, 0.53 mmol). Yield: 70%, LC-MS: [M+H] + =854.

[0290] Step 3: Under ice bath conditions, H2-2 (600 mg, 0.70 mmol) was dissolved in 10 mL of a mixed solution of trifluoroacetic acid and dichloromethane (v / v, 1 / 1). Under nitrogen protection, the reaction mixture was allowed to react at room temperature for 1 hour. The reaction was then stopped, and a saturated sodium bicarbonate solution was slowly added to adjust the pH to approximately 8. The mixture was extracted with ethyl acetate and concentrated under reduced pressure. The residue was separated by flash column chromatography to obtain a yellow solid H2 (320 mg, 0.49 mmol). LC-MS: [M+H] + =654. Yield: 70%.

[0291] H2 was purified by a chiral column: Unichiral CMD-5H to obtain the target compounds H2a (peak1) and H2b (peak2).

[0292] Following the synthetic routes of compounds H2a or H2b, and using similar skeletal structures, the following target molecules were synthesized.

[0293] *or * Represents a chiral site; * indicates no chiral separation was performed.

[0294]

[0295]

[0296]

[0297]

[0298]

[0299]

[0300]

[0301] Example 5:

[0302]

[0303] Step 1: In a 50 mL reaction flask, intermediate a5 (170 mg, 0.94 mmol) was dissolved in 10 mL of anhydrous THF, and potassium tert-butoxide (180 mg, 1.56 mmol) was added. The mixture was stirred at room temperature for 30 minutes to obtain solution S2. In another 50 mL reaction flask, intermediate a17 (400 mg, 0.78 mmol) was dissolved in 10 mL of anhydrous THF. Under ice bath conditions, the prepared solution S2 was slowly added, and stirring was continued for 1 hour. The reaction was then stopped. The reaction solution was poured into 100 mL of ice water, extracted with ethyl acetate, and the solvent was removed by vacuum distillation. The residue was purified by flash column chromatography (petroleum ether: ethyl acetate = 1:1) to give a pale yellow solid H11-1 (300 mg, 0.41 mmol). Yield: 44%, LC-MS: [M+H] + =734.

[0304] Step 2: Under nitrogen protection, H11-1 (300 mg, 0.41 mmol) and CuCN (150 mg, 1.64 mmol) were dissolved in 8 mL of anhydrous DMF. The reaction solution was reacted at 100 °C for 6 hours, then cooled to room temperature to stop the reaction. 50 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and the residue was separated by flash column chromatography (petroleum ether: ethyl acetate = 2:1) to give a yellow solid H11-2 (110 mg, 0.17 mmol). Yield: 42%, LC-MS: [M+H] + =633.

[0305] Step 3: Under nitrogen protection, H11-2 (110 mg, 0.17 mmol) was dissolved in 10 mL of anhydrous toluene, and intermediate a7 (94 mg, 0.22 mmol), Pd(DPEPhos)Cl2 (22 mg, 0.03 mmol), and cesium carbonate (160 mg, 0.48 mmol) were added sequentially. The reaction mixture was reacted at 110 °C for 6 hours under nitrogen protection. The reaction was then stopped, cooled to room temperature, and the solvent was removed by vacuum distillation. The residue was separated by flash column chromatography (petroleum ether: ethyl acetate = 1:1) to give a yellow solid H11-3 (90 mg, 0.11 mmol). Yield: 63%, LC-MS: [M+H] + =845.

[0306] Step 4: Under ice bath conditions, H11-3 (90 mg, 0.11 mmol) was dissolved in 8 mL of a mixed solution of trifluoroacetic acid and dichloromethane (v / v, 1 / 1). Under nitrogen protection, the reaction mixture was allowed to react at room temperature for 1 hour. The reaction was then stopped, and saturated sodium bicarbonate solution was slowly added to the system to adjust the pH to approximately 8. The mixture was extracted with ethyl acetate and concentrated under reduced pressure. The residue was separated by flash column chromatography to obtain a yellow solid H11 (30 mg, 0.05 mmol). LC-MS: [M+H] + =645. Yield: 43%.

[0307] 1 H NMR(400MHz,DMSO-d6)δ8.45(s,1H),8.26(s,2H),7.35(dd,J=8.4,5.2Hz,1 H),7.21(t,J=8.8Hz,1H),4.58(d,J=13.6Hz,1H),4.47(d,J=12.8Hz,1H),4. 34(s,2H),4.15(d,J=16.8Hz,2H),4.02(d,J=13.6Hz,2H),3.88(d,J=12.8Hz ,2H),3.52-3.16(m,4H),1.96-1.83(m,6H),1.23(s,2H),0.85–0.74(m,4H).

[0308] Following the synthetic route of compound H11, and using a similar skeletal structure, the following target molecule was synthesized.

[0309] *or * Represents a chiral site; * indicates no chiral separation was performed.

[0310]

[0311]

[0312]

[0313]

[0314] Example 6:

[0315]

[0316] Step 1: In a 50 mL reaction flask, intermediate a19 (150 mg, 0.55 mmol) was dissolved in 5 mL of anhydrous THF, and potassium tert-butoxide (93 mg, 0.82 mmol) was added. The mixture was stirred at room temperature for 30 minutes to obtain solution S3. In another 50 mL reaction flask, intermediate a14 (280 mg, 0.58 mmol) was dissolved in 5 mL of anhydrous THF. Under ice bath conditions, the prepared solution S3 was slowly added, and stirring was continued for 1 hour. The reaction was then stopped. The reaction solution was poured into 50 mL of ice water, extracted with ethyl acetate, and the solvent was removed by vacuum distillation. The residue was purified by flash column chromatography (petroleum ether: ethyl acetate = 1:1) to give a pale yellow solid H13-1 (205 mg, 0.28 mmol). Yield: 51%, LC-MS: [M+H] + =744.

[0317] Step 2: Under nitrogen protection, H13-1 (205 mg, 0.28 mmol) was dissolved in 6 mL of anhydrous toluene, followed by the sequential addition of intermediate a7 (200 mg, 0.50 mmol), Pd(DPEPhos)Cl2 (13 mg, 0.015 mmol), and cesium carbonate (175 mg, 0.54 mmol). The reaction mixture was reacted at 105 °C for 10 hours under nitrogen protection. The reaction was then stopped, cooled to room temperature, and the solvent was removed under reduced pressure. The residue was separated by flash column chromatography (petroleum ether: ethyl acetate = 1:2) to give a yellow solid H13-2 (120 mg, 0.13 mmol). Yield: 45%, LC-MS: [M+H] + =955.

[0318] Step 3: Under ice bath conditions, H13-2 (120 mg, 0.13 mmol) was dissolved in 6 mL of a mixed solution of trifluoroacetic acid and dichloromethane (v / v, 1 / 1). Under nitrogen protection, the reaction mixture was allowed to react at room temperature for 1 hour. The reaction was then stopped, and a saturated sodium bicarbonate solution was slowly added to adjust the pH to approximately 8. The mixture was extracted with ethyl acetate and concentrated under reduced pressure. The residue was separated by flash column chromatography to obtain a yellow solid H13 (32 mg). LC-MS: [M+H] + =671.

[0319] 1H NMR(400MHz, DMSO-d6)δ8.26(s,2H),7.97(d,J=11.2Hz,1H),7.38–7.09(m,2H),5.88–5.43(m,2H),4.70–4.37(m, 2H),4.29–4.00(m,6H),3.82-3.64(m,,4H),3.74–3.36(m,4H),1.93(m,4H),1.60-1.50(m,2H),1.40-1.20(m,2H).

[0320] Following the synthetic route of compound H13, and using a similar skeletal structure, the following target molecule was synthesized.

[0321] *or * Represents a chiral site; * indicates no chiral separation was performed.

[0322]

[0323] Example 7:

[0324]

[0325] Step 1: Under nitrogen protection, intermediate H1-1 (1.0 g, 1.59 mmol) was dissolved in 12 mL of methanol, followed by the addition of sodium methoxide (102 mg, 4.77 mmol). The reaction mixture was heated to 60 °C and reacted for 2 hours. The reaction was then stopped and cooled to room temperature. 40 mL of water was added to the system, and the mixture was extracted with ethyl acetate. The solvent was removed by vacuum distillation. The residue was separated by flash column chromatography to give a pale yellow solid P5-1 (305 mg, 0.48 mmol). Yield: 30%, LC-MS: [M+H] + =640.

[0326] Step 2: Under nitrogen protection, P5-1 (305 mg, 0.48 mmol) was dissolved in 5 mL of anhydrous toluene, followed by the sequential addition of intermediate a7 (270 mg, 0.67 mmol), Pd(DPEPhos)Cl2 (90 mg, 0.14 mmol), and anhydrous cesium carbonate (330 mg, 0.96 mmol). The reaction mixture was reacted at 105 °C for 8 hours under nitrogen protection. The reaction was then stopped, cooled to room temperature, and the solvent was removed under reduced pressure. The residue was separated by column chromatography (PE / EA, 1 / 2) to give a yellow solid P5-2 (327 mg, 0.38 mmol). Yield: 80%, LC-MS: [M+H] + =853.

[0327] Step 3: Under ice bath conditions, P5-2 (327 mg, 0.38 mmol) was dissolved in 4 mL of a mixed solution of trifluoroacetic acid and dichloromethane (v / v, 1 / 1). Under nitrogen protection, the reaction mixture was allowed to react at room temperature for 1 hour. The reaction was then stopped, and a saturated sodium bicarbonate solution was slowly added to adjust the pH to approximately 8. The mixture was extracted with ethyl acetate and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain the target compound P5 (74 mg, 0.11 mmol). Yield: 30%, LC-MS: [M+H] + =653.

[0328] 1 H NMR (400MHz, DMSO-d6) δ8.16 (s, 2H), 7.87 (d, J = 0.8Hz, 1H), 7.29 (dd, J = 8.4, 5.3H z,1H),7.22–7.11(m,1H),5.68-5.54(m,1H),4.28(dt,J=35.0,17.4Hz,2H),4.06 (dd,J=35.9,10.3Hz,2H),3.56(t,J=18.0Hz,4H),3.21(s,3H)3.18–3.08(m,2H), 3.03(s,1H),2.90–2.76(m,1H),2.19–1.75(m,6H),1.63(dd,J=20.6,9.9Hz,4H).

[0329] Following the synthetic route of compound P5, and using a similar skeletal structure, the following target molecule was synthesized.

[0330] *or * Represents a chiral site; * indicates no chiral separation was performed.

[0331]

[0332] Example 8:

[0333]

[0334] Step 1: Under nitrogen protection in an ice bath, intermediate H2-1 (500 mg, 0.78 mmol) and trifluoroethanol (156 mg, 1.56 mmol) were dissolved in 10 mL of tetrahydrofuran. Then, NaH (94 mg, 2.34 mmol) was added. The reaction mixture was heated to room temperature and reacted for 3 hours. The reaction was then stopped and cooled to room temperature. 30 mL of water was added to the system, and the mixture was extracted with ethyl acetate. The solvent was removed under reduced pressure. The residue was separated by flash column chromatography to give a pale yellow solid P7-1 (337 mg, 0.47 mmol). Yield: 60%, LC-MS: [M+H] + =722.

[0335] Step 2: Under nitrogen protection, P7-1 (305 mg, 0.42 mmol) was dissolved in 5 mL of anhydrous toluene, followed by the sequential addition of intermediate a7 (270 mg, 0.67 mmol), Pd(DPEPhos)Cl2 (90 mg, 0.14 mmol), and anhydrous cesium carbonate (330 mg, 0.96 mmol). The reaction mixture was reacted at 105 °C for 8 hours under nitrogen protection. The reaction was then stopped, cooled to room temperature, and the solvent was removed by vacuum distillation. The residue was separated by column chromatography (PE / EA, 1 / 2) to give a yellow solid P7-2 (313 mg, 0.34 mmol). Yield: 80%, LC-MS: [M+H] + =934.

[0336] Step 3: Under ice bath conditions, P7-2 (313 mg, 0.34 mmol) was dissolved in 5 mL of a mixed solution of trifluoroacetic acid and dichloromethane (v / v, 1 / 1). Under nitrogen protection, the reaction mixture was allowed to react at room temperature for 1 hour. The reaction was then stopped, and saturated sodium bicarbonate solution was slowly added to adjust the pH to approximately 8. The mixture was extracted with ethyl acetate and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain the target compound P7 (82 mg, 0.11 mmol). Yield: 33%, LC-MS: [M+H] + =734.

[0337] 1 H NMR (400MHz, DMSO-d6) δ8.02(s,2H),7.82(s,1H),7.20(dd,J=8.3,5.4Hz,1H),7.12(t,J=8.9Hz, 1H),5.24-5.10(m,1H),4.96–4.75(m,2H),4.24(dd,J=10.8,4.4Hz,2H),3.65(d,J=13.2Hz,4H), 3.51(d,J=12.1Hz,2H),2.84(dd,J=23.1,10.0Hz,2H),2.72–2.58(m,1H),2.48(s,1H),2.36(dt, J=15.9,9.7Hz,2H),2.21–1.76(m,2H),1.76–1.60(m,4H),0.66–0.60(m,2H),0.46–0.41(m,2H).

[0338] Example 9:

[0339]

[0340] Step 1: Under nitrogen protection in an ice bath, intermediate H2-1 (500 mg, 0.78 mmol) and 4-methoxybenzyl alcohol (215 mg, 1.56 mmol) were dissolved in 10 mL of tetrahydrofuran. Then, NaH (94 mg, 2.34 mmol) was added. The reaction mixture was heated to room temperature and reacted for 3 hours. The reaction was then stopped and cooled to room temperature. 30 mL of water was added to the system, and the mixture was extracted with ethyl acetate. The solvent was removed under reduced pressure. The residue was separated by flash column chromatography to give a pale yellow solid P8-1 (385 mg, 0.51 mmol). Yield: 65%, LC-MS: [M+H] + =760.

[0341] Step 2: Under nitrogen protection, P8-1 (319 mg, 0.42 mmol) was dissolved in 5 mL of anhydrous toluene, followed by the sequential addition of intermediate a7 (270 mg, 0.67 mmol), Pd(DPEPhos)Cl2 (90 mg, 0.14 mmol), and anhydrous cesium carbonate (330 mg, 0.96 mmol). The reaction mixture was reacted at 105 °C for 8 hours under nitrogen protection. The reaction was then stopped, cooled to room temperature, and the solvent was removed by vacuum distillation. The residue was separated by column chromatography (PE / EA, 1 / 2) to give a yellow solid P8-2 (122 mg, 0.13 mmol). Yield: 30%, LC-MS: [M+H] + =972.

[0342] Step 3: Under ice bath conditions, P8-2 (122 mg, 0.13 mmol) was dissolved in 5 mL of a mixed solution of trifluoroacetic acid and dichloromethane (v / v, 1 / 1). Under nitrogen protection, the reaction mixture was allowed to react at room temperature for 1 hour. The reaction was then stopped, and a saturated sodium bicarbonate solution was slowly added to adjust the pH to approximately 8. The mixture was extracted with ethyl acetate and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain the target compound P8 (19 mg, 0.03 mmol). Yield: 23%, LC-MS: [M+H] + =653.

[0343] 1H NMR (400MHz, DMSO-d6) δ8.02 (s, 2H), 7.82 (s, 1H), 7.20 (dd, J = 8.3, 5.4Hz, 1H), 7.12 (t, J = 8. 9Hz,1H),5.32-5.18(m,1H),4.24(dd,J=10.8,4.4Hz,2H),3.65(d,J=13.2Hz,4H),3.51(d,J =12.1Hz,2H),2.84(dd,J=23.1,10.0Hz,2H),2.72–2.58(m,1H),2.48(s,1H),2.36(dt,J=15 .9,9.7Hz,2H),2.21–1.76(m,2H),1.76–1.60(m,4H),0.68–0.60(m,2H),0.48–0.41(m,2H).

[0344] Example 10:

[0345]

[0346] Step 1: Under nitrogen protection in an ice bath, intermediates a3 (900 mg, 2.1 mmol) and a5 (360 mg, 2.1 mmol) were dissolved in 15 mL of tetrahydrofuran. Then, NaH (100 mg, 4.2 mmol) was added. The reaction mixture was heated to 40 °C and reacted for 3 hours. The reaction was then stopped and cooled to room temperature. 30 mL of water was added to the system, and the mixture was extracted with ethyl acetate. The solvent was removed by vacuum distillation. The residue was separated by flash column chromatography to give a yellow solid P9-2 (490 mg, 0.81 mmol). Yield: 38%, LC-MS: [M+H] + =565.

[0347] Step 2: Under nitrogen protection, P9-2 (440 mg, 0.78 mmol) was dissolved in 6 mL of a mixed solution of 1,4-dioxane and water (v / v, 5 / 1). Then, starting material P9-1 (480 mg, 0.94 mmol), Xphos Pd G1 (180 mg, 0.23 mmol), and anhydrous cesium carbonate (1.02 g, 3.12 mmol) were added sequentially. The reaction mixture was reacted at 95 °C for 3 hours under nitrogen protection. The reaction was then stopped, cooled to room temperature, and the solvent was removed by vacuum distillation. The residue was separated by column chromatography (PE / EA, 1 / 2) to give a yellow solid P9-3 (500 mg, 0.55 mmol). Yield: 70%, LC-MS: [M+H] + =915.

[0348] Step 3: Dissolve intermediate P9-3 (500 mg, 0.55 mmol) from the previous step in 20 mL of tetrahydrofuran, and add tetrabutylammonium fluoride (220 mg, 0.9 mmol). React at room temperature for 12 hours, monitoring the reaction until complete by TLC. Dissolve the reaction solution in 50 mL of water, extract with ethyl acetate, dry to anhydrous sodium sulfate, and concentrate to obtain crude P9-4 (400 mg). LC-MS: [M+H] + =759.

[0349] Step 4: Under ice bath conditions, crude P9-4 (400 mg, 0.53 mmol) was dissolved in 5 mL of a mixed solution of trifluoroacetic acid and dichloromethane (v / v, 1 / 1). Under nitrogen protection, the reaction mixture was allowed to react at room temperature for 1 hour. The reaction was then stopped, and saturated sodium bicarbonate solution was slowly added to adjust the pH to approximately 8. The mixture was extracted with ethyl acetate and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain the target compound P9 (45 mg, 0.07 mmol). Yield: 14%, LC-MS: [M+H] + =615.

[0350] 1 H NMR (400MHz, DMSO-d6) δ10.30(s,1H),9.10(s,1H),7.98(dd,J=9.2,6.0Hz,1H),7.47(t,J=9.0Hz,1H),7.41(d,J=2.4Hz,2H),7.37(s,1H ),7.24(s,1H),7.22(s,2H),7.11(s,1H),4.65(d,J=13.7Hz,1H),4.49(s,1H),4.35(s,3H),4.18(s,4H),4.0-3.90(m,6H),1.97(m,7H).

[0351] Following the synthetic route of compound P9, and using a similar skeletal structure, the following target molecule was synthesized.

[0352]

[0353] Example 11:

[0354]

[0355] Step 1: In a 20 mL reaction flask, intermediate a5 (142 mg, 0.82 mmol) was dissolved in 5 mL of anhydrous THF, and potassium tert-butoxide (138 mg, 2.34 mmol) was added. The mixture was stirred at room temperature for 30 minutes to obtain solution S4. In another 20 mL reaction flask, intermediate a27 (400 mg, 0.82 mmol) was dissolved in 5 mL of anhydrous THF. Under ice bath conditions, the prepared solution S4 was slowly added, and stirring was continued for 1 hour. The reaction was then stopped. The reaction solution was poured into 100 mL of ice water, extracted with ethyl acetate, and the solvent was removed by vacuum distillation. The residue was purified by flash column chromatography (petroleum ether: ethyl acetate = 1:1) to give a pale yellow solid P12-1 (420 mg, 0.66 mmol). Yield: 80%, LC-MS: [M+H] + =642.

[0356] Step 2: Under nitrogen protection, P12-1 (420 mg, 0.66 mmol) was dissolved in 5 mL of anhydrous toluene, followed by the sequential addition of intermediate a7 (352 mg, 0.87 mmol), Pd(DPEPhos)Cl2 (22 mg, 0.03 mmol), and cesium carbonate (438 mg, 1.35 mmol). The reaction mixture was reacted at 105 °C for 10 hours under nitrogen protection. The reaction was then stopped, cooled to room temperature, and the solvent was removed under reduced pressure. The residue was separated by flash column chromatography (petroleum ether: ethyl acetate = 1:2) to give a yellow solid P12-2 (450 mg, 0.53 mmol). Yield: 80%, LC-MS: [M+H] + =854.

[0357] Step 3: Under ice bath conditions, P12-2 (450 mg, 0.53 mmol) was dissolved in 6 mL of a mixed solution of trifluoroacetic acid and dichloromethane (v / v, 1 / 1). Under nitrogen protection, the reaction mixture was allowed to react at room temperature for 1 hour. The reaction was then stopped, and a saturated sodium bicarbonate solution was slowly added to adjust the pH to approximately 8. The mixture was extracted with ethyl acetate and concentrated under reduced pressure. The residue was separated by flash column chromatography to obtain a yellow solid P12 (104 mg, 0.16 mmol). LC-MS: [M+H] + =654. Yield: 30%.

[0358] 1H NMR (400MHz, DMSO-d6) δ8.15(s,2H),7.81(s,1H),7.26(dd,J=8.4,5.3Hz,1H),7.15(t,J=8.9Hz,1H),5.24–5.10(m,1H),4.22(dt,J=19.4,1 0.7Hz,2H),3.86–3.62(m,4H),3.01(s,2H),2.87–2.75(m,2H),2.42- 2.33(m,3H),2.20–1.95(m,2H),1.91–1.75(m,1H),0.67–0.38(m,8H).

[0359] Following the synthetic route of compound P12, and using a similar skeletal structure, the following target molecule was synthesized.

[0360]

[0361] Example 12:

[0362]

[0363] Step 1: In a high-pressure reactor, the starting material 4-bromo-2,6-difluorobenzonitrile a5 (100 g, 459 mmol) was dissolved in 240 mL of ethanol, and 400 mL of ammonia water was added. The mixture was heated to 90 °C and reacted for 16 hours. After cooling to room temperature, the reaction was stopped. The solvent was removed by vacuum distillation, and 100 mL of ice water was added to the reaction solution. The mixture was extracted with ethyl acetate, and the solvent was removed by vacuum distillation. The residue was purified by flash column chromatography (petroleum ether / ethyl acetate, 9 / 1) to give a yellow solid P14-2 (74.9 g, 352 mmol). Yield: 77%, LC-MS: [M+H] + =215.

[0364] Step 2: Under nitrogen protection, 33.8 g (252 mmol) of 2,2-diethoxyethanol was dissolved in 450 mL of anhydrous DMF. NaH (10.08 g, 252 mmol) was slowly added at 0 °C. After stirring for 1 hour, intermediate P14-2 (45 g, 210 mmol) from the previous step was added. The ice bath was removed, and the mixture was heated to 50 °C and reacted for 2 hours. The reaction was then stopped, cooled to room temperature, and 1 L of water was added to the system. The mixture was extracted with ethyl acetate, and the solvent was removed under reduced pressure. The residue was separated by flash column chromatography (petroleum ether / ethyl acetate, 10 / 1) to give a yellow solid, P14-3 (25.3 g, 77.1 mmol). Yield: 37%, LC-MS: [M+H] + =329.

[0365] Step 3: Add 100 mL of toluene to polyphosphoric acid (10.42 g), heat to 100 °C, add intermediate P14-3 (10.0 g, 30.4 mmol) from the previous step, and continue the reaction at this temperature for 2 hours. Then stop the reaction. Slowly pour the reaction solution into a large amount of ice water, extract with ethyl acetate, and concentrate under reduced pressure. Separate the residue by flash column chromatography (petroleum ether / ethyl acetate, 10 / 1) to give a yellow solid P14-4 (1.04 g, 4.4 mmol), yield: 14%. LC-MS: [M+H] + =236.

[0366] Step 4: Dissolve the intermediate P14-4 (8.3 g, 35.0 mmol) from the previous step in 150 mL of ethanol, add 7.94 g of KOH aqueous solution (50 mL), and react at 90 °C for 4 hours. Stop the reaction. Remove the organic solvent under reduced pressure, add 100 mL of ice water to the reaction mixture, extract with dichloromethane, and concentrate under reduced pressure. Separate the residue by flash column chromatography (petroleum ether / dichloromethane, 2 / 1) to give a yellow solid P14-5 (4.0 g, 15.7 mmol), yield: 45%. LC-MS: [M+H] + =256.

[0367] Step 5: Under nitrogen protection, the intermediate P14-5 (3.3 g, 13.0 mmol) from the previous step was dissolved in 33 mL of anhydrous THF. A tetrahydrofuran solution (20 mL) containing triphosgene (3.6 g, 12.4 mmol) was slowly added dropwise at 0 °C. The reaction was allowed to proceed to room temperature for 2 hours. The reaction was then stopped, and 100 mL of water was added to the system. The mixture was extracted with ethyl acetate, and the solvent was removed under reduced pressure to obtain crude P14-6 (2.8 g). LC-MS: [M+H] + =282.

[0368] Step 6: Under nitrogen protection, the crude product P14-6 (2.8 g) from the previous step was dissolved in 50 mL of phosphorus oxychloride, and 5 mL of N,N-diisopropylethylamine was added. The mixture was heated to 100 °C and reacted for 2 hours. The solvent was removed by vacuum distillation, and 100 mL of water was added to the system. The mixture was extracted with ethyl acetate, concentrated, and the residue was separated by flash column chromatography (petroleum ether / ethyl acetate, 3 / 1) to give a yellow solid P14-7 (1.1 g, 3.5 mmol). Two-step yield: 27%. LC-MS: [M+H] + =319.

[0369] Step 7: At room temperature, intermediate P14-7 (1.0 g, 3.15 mmol) and starting material 3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester a2-1 (670 mg, 3.15 mmol) were dissolved in 20 mL of 1,4-dioxane. N,N-diisopropylethylamine (1.7 mL, 9.5 mmol) was added, and the mixture was heated to 50 °C and reacted for 2 hours. 60 mL of water was added to the system, and the mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by column chromatography to obtain a white solid P14-8 (900 mg, 1.82 mmol). Yield: 58%. LC-MS: [M+H] + =495.

[0370] Step 8: Under nitrogen protection, intermediate P14-8 (520 mg, 1.05 mmol) and cesium carbonate (684 mg, 2.10 mmol) were dissolved in 5 mL of DMF, and intermediate a5 (363 mg, 2.10 mmol) was added. The mixture was heated to 140 °C and reacted for 2 hours. After cooling to room temperature, 60 mL of water was added to the system, and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by column chromatography (petroleum ether / ethyl acetate, 1 / 1) to give a yellow solid P14-9 (155 mg, 0.25 mmol). Yield: 23%. LC-MS: [M+H] + =630.

[0371] Step 9: Under nitrogen protection, intermediate P14-9 (155 mg, 0.25 mmol) and cesium carbonate (200 mg, 0.62 mmol) were dissolved in 5 mL of toluene. Intermediate a7 (139 mg, 0.34 mmol) and Pd(DPEPhos)Cl2 (52 mg, 0.074 mmol) were added, and the mixture was heated to 105 °C and reacted for 3 hours. After cooling to room temperature, 30 mL of water was added, and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by column chromatography (petroleum ether / ethyl acetate, 1 / 4) to give a yellow solid P14-10 (85 mg, 0.25 mmol). Yield: 41%. LC-MS: [M+H] + =842.

[0372] Step 10: Under ice bath conditions, P14-10 (65 mg, 0.077 mmol) was dissolved in 3 mL of a mixed solution of trifluoroacetic acid and dichloromethane (v / v, 1 / 1). Under nitrogen protection, the reaction mixture was allowed to react at room temperature for 1 hour. The reaction was then stopped, and saturated sodium bicarbonate solution was slowly added to the system to adjust the pH to approximately 8. The mixture was extracted with ethyl acetate and concentrated under reduced pressure. The residue was separated by preparative HPLC (X Bridge Shield RP18 OBD column, 19*150 mm, 5 μm; mobile phase A: water (10 mmol / L NH4HCO3), mobile phase B: acetonitrile; flow rate: 25 mL / min) to obtain a white solid P14 (9.2 mg). LC-MS: [M+H] + =642.

[0373] 1 H NMR(400MHz,DMSO-d6)δ8.11(d,J=4.0Hz,1H),7.97(brs,2H),7.36-7.31(m,2H),7.17-7.11 (m,1H),6.57(d,J=4.0Hz,1H),5.17(d,J=56.0Hz,1H),4.31-4.18(m,2H),4.07-4.03(m,1H), 3.93-3.89(m,1H),3.49-3.45(m,2H),2.89-2.73(m,3H),2.41-2.27(m,3H),2.17-2.08(m,2 H),1.93-1.79(m,3H),1.66-1.62(m,2H),1.32(s,1H),0.64-0.61(m,2H),0.45-0.42(m,2H).

[0374] Example 13:

[0375]

[0376] Step 1: Using an ice bath, dissolve intermediate c1 (330 mg, 1.76 mmol) in 1 mL of anhydrous tetrahydrofuran, add NaH (85 mg, 3.52 mmol), heat to room temperature and stir for 0.5 hours. Set the reaction solution aside. Dissolve intermediate a3 (350 mg, 0.82 mmol) in 1 mL of anhydrous tetrahydrofuran, add the above reaction solution, and react at room temperature for 1 hour. Stop the reaction. Concentrate under reduced pressure, and separate the crude product by flash column chromatography to obtain compound P18-1 (240 mg, 0.42 mmol), yield 24%. LC-MS: [M+H] + =579.

[0377] Step 2: Under nitrogen protection, compound P18-1 (240 mg, 0.42 mmol), intermediate b2 (270 mg, 0.44 mmol), cesium carbonate (270 mg, 0.83 mmol), and methanesulfonic acid [n-butyldi(1-adamantyl)phosphine] (2-amino-1,1'-biphenyl-2-yl)palladium (Pd-G3, 10 mg, 0.014 mmol) were dissolved in 2 mL of a mixed solution of 1,4-dioxane and water (v / v, 5 / 1). The mixture was heated to 95 °C and reacted for 1 hour, after which the reaction was stopped. The solvent was removed under reduced pressure, and the crude product was separated by flash column chromatography to obtain compound P18-2 (300 mg, 0.29 mmol), with a yield of 70%. LC-MS: [M+H] + =1041.

[0378] Step 3: Compound P18-2 (270 mg, 0.26 mmol) and cesium fluoride (80 mg, 0.52 mmol) were dissolved in 1 mL of DMF, and the mixture was heated to 50 °C and reacted for 1 hour. The reaction was then stopped. 5 mL of water was added to the system, and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain crude P18-3. LC-MS: [M+H] + =729.

[0379] Step 4: Dissolve the crude product P18-3 from the previous step in 1 mL of 1,4-dioxane hydrogen chloride solution (4M concentration), react at room temperature for 1 hour, slowly add saturated sodium bicarbonate aqueous solution to adjust the pH to approximately 8, extract with ethyl acetate, concentrate under reduced pressure, and purify by TLC (thin-layer chromatography) to obtain compound P18 (50 mg). LC-MS: [M+H] + =629.

[0380] 1H NMR (400MHz, DMSO-d6) δ10.17(s,1H),9.06(s,1H),7.99(dd,J=9.2,5.9Hz,1H),7.48(t,J=9.0Hz,1H),7.41(d,J=2 .5Hz,1H),7.19(t,J=2.2Hz,1H),4.49(d,J=11.4Hz,1H),4.33(d,J=12.6Hz,1H),4.12(dd,J=10.5,3.4Hz,1H),4.0 2(dd,J=10.6,2.7Hz,1H),3.95(s,1H),3.69–3.53(m,4H),3.45–3.25(m,2H),2.95–2.85(m,1H),2.72–2.64(m,1H) ,2.57–2.52(m,1H),2.47–2.32(m,2H),2.09(s,3H),1.95–1.72(m,4H),1.90–1.60(m,4H),1.51(t,J=11.7Hz,1H).

[0381] Following the synthetic route of compound P18, and using a similar skeletal structure, the following target molecule was synthesized.

[0382]

[0383]

[0384]

[0385] Example 14:

[0386]

[0387] Under ice bath conditions, starting material P29-1 (305 mg, 2.4 mmol) and intermediate a3 (500 mg, 1.98 mmol) were dissolved in 10 mL of dichloromethane. Triethylamine (301 mg, 2.97 mmol) was added, and the mixture was reacted at room temperature for 2 hours. 30 mL of water was added, and the mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by column chromatography to give a white solid P29-2 (650 mg, 1.9 mmol), yield: 95%. LC-MS: [M+H] + =344.

[0388] In a 50 mL reaction flask, compound P29-2 (300 mg, 0.87 mmol) was dissolved in 6 mL of anhydrous tetrahydrofuran in an ice bath. Sodium hydride (70 mg, 1.74 mmol) was added, and the mixture was stirred for 30 minutes to obtain solution S1. Intermediate a18 was added to solution S1, and the mixture was heated to 70 °C and reacted for 16 hours. The reaction was then stopped by cooling to room temperature. The reaction was quenched with 30 mL of ice water, extracted with ethyl acetate, and concentrated under reduced pressure to remove the solvent. The residue was purified by flash column chromatography (DCM / MeOH = 10 / 1) to give a white solid P29-3 (200 mg, 0.40 mmol), yield: 46%. LC-MS: [M+H] + =495.

[0389] Under nitrogen protection, compound P29-3 (450 mg, 0.91 mmol) and starting material P9-1 (560 mg, 1.1 mmol) were dissolved in 10 mL of 1,4-dioxane and 2 mL of water. Catalyst XPhos Pd G2 (215 mg, 0.3 mmol) and cesium carbonate (1.18 g, 3.64 mmol) were added sequentially. The reaction solution was reacted at 95 °C for 2 hours, after which the reaction was stopped. The solvent was removed by concentration under reduced pressure, and the residue was separated by flash column chromatography (DCM / MeOH = 20 / 1) to give a yellow solid P29-4 (500 mg, 0.59 mmol). Yield: 65%, LC-MS: [M+H]+ = 845.

[0390] Compound P29-4 (200 mg, 0.24 mmol) was dissolved in 4 mL of tetrahydrofuran at room temperature, and tetrabutylammonium fluoride (TBAF, 130 mg, 0.48 mmol) was added. The mixture was reacted at room temperature for 1 hour. 20 mL of water was added to the system, and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and concentrated to give crude yellow oil P29-5 (160 mg, 0.23 mmol). Yield: 98%. LC-MS: [M+H] + =689.

[0391] The crude P29-5 (163 mg, 0.24 mmol) from the previous step was dissolved in 2 mL of dichloromethane, and 1 mL of a 1,4-dioxane solution of hydrogen chloride (4N concentration) was added dropwise. The reaction was carried out at room temperature for 1 hour, and then stopped. A saturated sodium bicarbonate solution was slowly added to the system to adjust the pH to approximately 8. The mixture was extracted with ethyl acetate and concentrated under reduced pressure. The residue was separated by flash column chromatography to give 10 mg of yellow solid P29. LC-MS: [M+H] + =645.

[0392] 1H NMR (400MHz, DMSO-d6) δ10.16(s,1H),9.34(s,1H),8.77(s,1H),8.01(dd,J=9. 2,5.9Hz,1H),7.49(t,J=9.0Hz,1H),7.42(d,J=2.4Hz,1H),7.19(t,J=3.5Hz,1 H),5.30–5.02(m,1H),4.54–4.41(m,2H),4.22-4.05(m,1H),4.00-3.92(m,1H) ,3.87-3.68(m,1H),2.85–2.60(m,5H),2.42-2.20(m,7H),2.16–1.67(m,14H).

[0393] Example 15:

[0394]

[0395] Step 1: Dissolve intermediate d2 (1.4 g, 1.85 mmol) in 10 mL of tetrahydrofuran, and slowly add 5.0 mL of saturated lithium hydroxide aqueous solution. React at room temperature for 40 minutes, then stop the reaction. Slowly add 1 M dilute hydrochloric acid to adjust the pH to approximately 5, extract with ethyl acetate, concentrate, and obtain crude product M1-1, which can be used directly in the next step of the reaction. LC-MS: [M+H] + =727.

[0396] Step 2: Ice bath. Dissolve the crude product M1-1 from the previous step in a mixed solution of trifluoroacetic acid and dichloromethane (4.0 mL, 1 / 1). React in an ice bath for 1 hour, then stop the reaction. Remove the solvent under reduced pressure. Separate the crude product by HPLC preparative chromatography to obtain compound M1 (10 mg). LC-MS: [M+H] + =527.

[0397] 1H NMR (400MHz, DMSO-d6) δ8.10(s,2H),7.81(s,1H),7.32–7.21(m,1H),7.18–7.10(m,1H),4.27(d,J=10.7Hz,2H),3.47(s,4H),1.58(s,4H).

[0398] Example 16:

[0399]

[0400] Step 1: In an ice bath, compound M1-1 (400 mg, 0.55 mmol), starting material M2-1 (109 mg, 0.83 mmol), and DIEA (213 mg, 1.65 mmol) were dissolved in 10 mL of DMF. After stirring for 5 minutes, HATU (315 mg, 0.83 mmol) was added, and the mixture was reacted at room temperature for 2 hours. The reaction solution was poured into 150 mL of ice water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by column chromatography to obtain a colorless oily substance M2-2 (393 mg, 0.47 mmol), with a yield of 85%. LC-MS: [M+H] + =837

[0401] Step 2: Dissolve intermediate M2-2 (393 mg, 0.47 mmol) from the previous step in 4.0 mL of dichloromethane, add 1.0 mL of trifluoroacetic acid, and react at room temperature for 1 hour. Stop the reaction. Remove the solvent under reduced pressure, dissolve the crude product in 10 mL of water, adjust the pH to approximately 8 with saturated sodium bicarbonate solution, extract with ethyl acetate, dry to anhydrous sodium sulfate, concentrate, and separate the crude product by column chromatography to obtain compound M2 (89 mg, 0.14 mmol), yield: 30%. LC-MS: [M+H] + =637.

[0402] 1 H NMR(400MHz,DMSO-d6)δ8.5(d,J=8.4Hz,1H),8.12(s,2H),7.97(s,1H),7.33–7.25( m,1H),7.17(t,J=8.9Hz,1H),4.49(t,J=11.4Hz,1H),4.37(t,J=10.4Hz,1H),4.17(t ,J=7.5Hz,1H),3.68-3.51(m,5H),2.86–2.80(m,1H),2.68–2.66(m,1H),2.34–2.31 (m,1H),2.02–1.92(m,1H),1.84–1.75(m,1H),1.71–1.46(m,9H),1.25–1.22(m,2H).

[0403] Following the synthetic route of compound M2, and using a similar skeletal structure, the following target molecule was synthesized.

[0404] * indicates a chiral molecule that has not undergone chiral resolution.

[0405]

[0406]

[0407] Example 17:

[0408]

[0409] Step 1: Under nitrogen protection, in a 50 mL reaction flask, intermediate a31 (300 mg, 1.0 mmol) was dissolved in 5 mL of anhydrous THF, and potassium tert-butoxide (224 mg, 2.0 mmol) was added. The mixture was stirred at room temperature for 30 minutes to obtain solution S5. In another 50 mL reaction flask, intermediate a14 (488 mg, 1.0 mmol) was dissolved in 5 mL of anhydrous THF. Under ice bath conditions, the prepared solution S5 was slowly added, and stirring was continued for 1 hour. The reaction was then stopped. The reaction solution was poured into 50 mL of ice water, extracted with ethyl acetate, and the solvent was removed by vacuum distillation. The residue was purified by flash column chromatography (petroleum ether: ethyl acetate = 1:1) to give a pale yellow solid H23-1 (240 mg, 0.32 mmol). Yield: 31%, LC-MS: [M+H] + =757.

[0410] Step 2: Under nitrogen protection, compound H23-1 (240 mg, 0.32 mmol) was dissolved in 6 mL of anhydrous toluene, followed by the sequential addition of intermediate a7 (188 mg, 0.45 mmol), Pd(DPEPhos)Cl2 (46 mg, 0.064 mmol), and cesium carbonate (209 mg, 0.64 mmol). The reaction mixture was reacted at 105 °C for 10 hours under nitrogen protection. The reaction was then stopped, cooled to room temperature, and the solvent was removed under reduced pressure. The residue was separated by flash column chromatography (petroleum ether: ethyl acetate = 1:2) to give a yellow solid H23-2 (250 mg, 0.26 mmol). Yield: 79%, LC-MS: [M+H] + =982.

[0411] Step 3: Under ice bath conditions, compound H23-2 (250 mg, 0.26 mmol) was dissolved in 6 mL of a mixed solution of trifluoroacetic acid and dichloromethane (v / v, 1 / 1). Under nitrogen protection, the reaction mixture was allowed to react at room temperature for 1 hour. The reaction was then stopped, and a saturated sodium bicarbonate solution was slowly added to adjust the pH to approximately 8. The mixture was extracted with ethyl acetate and concentrated under reduced pressure. The residue was separated by flash column chromatography to obtain a yellow solid H23 (10 mg). LC-MS: [M+H] + =698.

[0412] 1H NMR(400MHz, DMSO-d6)δ8.16(s,2H),7.87(s,1H),7.28(dd,J=8.4,5.3Hz,1H),7.17(t,J=8.9Hz,1H),5.19–5.05(m,1H),4.38–4.2 9(m,5H),3.68–3.33(m,9H),3.18–3.09(m,1H),3.01(d,J=13.1Hz,1H),2.88-2.85(m,1H),2.67–2.56(m,2H),2.09-1.53(m,12H).

[0413] Example 18:

[0414]

[0415] Step 1: In a 50 mL reaction flask, intermediate a18 (176 mg, 0.94 mmol) was dissolved in 10 mL of anhydrous THF, and potassium tert-butoxide (180 mg, 1.56 mmol) was added. The mixture was stirred at room temperature for 30 minutes to obtain solution S6. In another 50 mL reaction flask, intermediate a17 (400 mg, 0.78 mmol) was dissolved in 10 mL of anhydrous THF. Under ice bath conditions, the prepared solution S2 was slowly added, and stirring was continued for 1 hour. The reaction was then stopped. The reaction solution was poured into 100 mL of ice water, extracted with ethyl acetate, and the solvent was removed by vacuum distillation. The residue was purified by flash column chromatography (petroleum ether: ethyl acetate = 1:1) to give a pale yellow solid H24-1 (337 mg, 0.45 mmol). Yield: 48%, LC-MS: [M+H] + =749.

[0416] Step 2: Under nitrogen protection, H24-1 (1.0 g, 1.34 mmol), potassium carbonate, and methylboric acid (80 mg, 1.34 mmol) were dissolved in 10 mL of anhydrous DMA. Pd(PPh3)Cl2 (100 mg, 0.13 mmol) was added. The reaction mixture was reacted at 90 °C for 2 hours, then cooled to room temperature to stop the reaction. 50 mL of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated. The residue was separated by flash column chromatography (petroleum ether:ethyl acetate = 3:1) to give a yellow solid H24-2 (340 mg, 0.54 mmol). Yield: 40%, LC-MS: [M+H] + =636.

[0417] Step 3: Under nitrogen protection, intermediate H24-2 (340 mg, 0.54 mmol) from the previous step was dissolved in 10 mL of anhydrous toluene. Intermediate a7 (900 mg, 2.2 mmol), Pd(DPEPhos)Cl2 (170 mg, 0.24 mmol), and cesium carbonate (1.0 g, 3.2 mmol) were added sequentially. The reaction mixture was reacted at 110 °C for 6 hours under nitrogen protection. The reaction was then stopped, cooled to room temperature, and the solvent was removed under reduced pressure. The residue was separated by flash column chromatography (petroleum ether: ethyl acetate = 1:1) to give a yellow solid H24-3 (300 mg, 0.35 mmol). Yield: 65%, LC-MS: [M+H] + =848.

[0418] Step 4: Under ice bath conditions, dissolve H24-3 (300 mg, 0.35 mmol) in 3 mL of a mixed solution of trifluoroacetic acid and dichloromethane (v / v, 1 / 1). Under nitrogen protection, react the solution at room temperature for 1 hour. Stop the reaction, slowly add saturated sodium bicarbonate solution to the system and adjust the pH to approximately 8. Extract with ethyl acetate and concentrate under reduced pressure. Separate the residue by preparative HPLC to obtain a yellow solid H24 (10 mg). LC-MS: [M+H] + =648.

[0419] 1 H NMR(400MHz,DMSO-d6)δ8.03(s,2H),7.61(s,1H),7.22–7.08(m,2H),5.24-5.10(m,1H),4.38(s,2H),4.27(dd, J=25.6,12.1Hz,2H),3.53–3.42(m,5H),2.80–2.62(m,5H),2.40–2.30(m,1H),2.07(s,3H),2.01–1.66(m,12H).

[0420] Example 19:

[0421]

[0422] Step 1: Under nitrogen protection at -25°C, compound H10-1 (1.1 g, 1.68 mmol) obtained according to the route in Example 4 was dissolved in 20 mL of anhydrous tetrahydrofuran. Isopropyl magnesium bromide (5.0 mL, 5.0 mmol) was slowly added dropwise. After the addition was complete, isopropanol pinacol borate (1.56 g, 8.4 mmol) was added to the reaction solution. The reaction was stopped at -25°C for 2 hours. The reaction was quenched by adding 20 mL of saturated ammonium chloride aqueous solution to the mixture. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by flash column chromatography (petroleum ether / ethyl acetate = 2 / 1) to give a pale yellow solid H26-1 (800 mg, 1.28 mmol), yield: 76%. LC-MS: [M+H] + =622.

[0423] Step 2: Under nitrogen protection, compound H26-1 (250 mg, 0.74 mmol), cesium carbonate (480 mg, 1.48 mmol), and intermediate b3 (690 mg, 1.11 mmol) were dissolved in 10 mL of anhydrous toluene. The catalyst bis(diphenylphosphine ether)palladium dichloride (100 mg, 0.15 mmol) was added. The reaction solution was reacted at 105 °C for 3 hours. The reaction was then stopped, and the mixture was filtered. The solvent was removed by vacuum distillation. The crude product was separated by flash column chromatography (petroleum ether / ethyl acetate = 1 / 2) to give a pale yellow solid H26-2 (160 mg, 0.19 mmol), yield: 26%. LC-MS: [M+H] + =835.

[0424] Step 3: Under ice bath conditions, compound H26-2 (160 mg, 0.19 mmol) was dissolved in 5 mL of a mixed solution of trifluoroacetic acid and dichloromethane (v / v, 1 / 1). The reaction was carried out at room temperature for 1 hour. The reaction was then stopped, and saturated sodium bicarbonate solution was slowly added to the system to adjust the pH to approximately 8. The mixture was extracted with ethyl acetate and concentrated under reduced pressure. The residue was separated by flash column chromatography to obtain a yellow solid H26 (20 mg, 0.032 mmol). LC-MS: [M+H] + =635.

[0425] 1HNMR(400MHz,DMSO-d6)δ7.85(s,1H),7.55(d,J=8.6Hz,1H),6.55(d,J=8.6Hz,1H),6.39–6.38(m,2H),5.11(dt,J=9.4,5.1Hz,1H),4.42(s,2H) ,4.29(d,J=12.3Hz,2H),3.70–3.62(m,6H),2.81–2.66(m,3H),2.65–2. 57(m,3H),2.33(q,J=8.2Hz,1H),2.10–1.80(m,9H),1.75–1.65(m,5H).

[0426] Example 20:

[0427]

[0428] Step 1: Under nitrogen protection at -25°C, compound H10-1 (100 mg, 0.15 mmol), hexamethyldistin (74 mg, 0.22 mmol), tricyclohexylphosphine (8.4 mg, 0.03 mmol), and lithium chloride (19 mg, 0.45 mmol) obtained according to the route in Example 4 were dissolved in 10 mL of 1,4-dioxane. Catalyst Pd2dba3 (27 mg, 0.03 mmol) was added, and the mixture was heated to 95°C and reacted for 12 hours. The reaction was then stopped. 40 mL of ice water was added to the mixture, and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by flash column chromatography (petroleum ether / ethyl acetate = 2 / 1) to give a yellow solid H32-1 (80 mg, 0.11 mmol), yield: 72%. LC-MS: [M+H] + =742.

[0429] Step 2: Under nitrogen protection, compound H32-1 (80 mg, 0.11 mmol), cesium carbonate (110 mg, 0.33 mmol), and intermediate b7 (25 mg, 0.12 mmol) were dissolved in 10 mL of 1,4-dioxane. The catalyst bis(triphenylphosphine)palladium dichloride (15 mg, 0.022 mmol) was added. The reaction solution was reacted at 100 °C for 4 hours, then the reaction was stopped and filtered. The solvent was removed by vacuum distillation. The crude product was separated by flash column chromatography (petroleum ether / ethyl acetate = 2 / 1) to give a pale yellow solid H32-2 (20 mg, 0.03 mmol), yield: 24%. LC-MS: [M+H] + =750.

[0430] Step 3: Under ice bath conditions, compound H32-2 (20 mg, 0.03 mmol) was dissolved in 1 mL of a mixed solution of trifluoroacetic acid and dichloromethane (v / v, 1 / 1). The reaction was carried out at room temperature for 1 hour. The reaction was then stopped, and saturated sodium bicarbonate solution was slowly added to the system to adjust the pH to approximately 8. The mixture was extracted with ethyl acetate and concentrated under reduced pressure. The residue was separated by flash column chromatography to obtain a yellow solid H32 (2.1 mg). LC-MS: [M+H] + =650.

[0431] 1 H NMR(400MHz,DMSO-d6)δ7.98(s,1H),6.67(s,2H),5.19–5.05(m,1H),4.42(s,2H),4.29(d,J=12.3Hz ,2H),3.70–3.62(m,6H),2.81–2.66(m,3H),2.65–2.57(m,3H),2.33–1.80(m,9H),1.75–1.65(m,5H).

[0432] Example 21:

[0433]

[0434] Step 1: Under nitrogen protection at -60°C, compound H24-1 (1.0 g, 1.34 mmol) obtained from the route in Example 18 was dissolved in 20 mL of anhydrous tetrahydrofuran. Isopropyl magnesium bromide (1.3 mL, 1.3 mmol) was slowly added dropwise. After stirring for 30 minutes, anhydrous DMF (196 mg, 2.38 mmol) was added to the reaction mixture, and the mixture was stirred again for 30 minutes before stopping the reaction. The reaction was quenched by adding 20 mL of saturated ammonium chloride aqueous solution to the mixture. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to obtain crude product H33-1. LC-MS: [M+H] + =650.

[0435] Step 2: Ice bath. Dissolve the crude product H33-1 from the previous step in 10 mL of methanol, slowly add sodium borohydride (102 mg, 2.68 mmol), heat to room temperature and react for 1 hour, then stop the reaction. Add 30 mL of saturated ammonium chloride aqueous solution to the reaction solution, extract with ethyl acetate, dry to anhydrous sodium sulfate, concentrate, and separate the crude product by flash column chromatography (petroleum ether / ethyl acetate = 2 / 1) to obtain a pale yellow solid H33-2 (200 mg, 0.31 mmol). Two-step yield: 23%. LC-MS: [M+H] + =652.

[0436] Step 3: Under nitrogen protection, intermediate H33-2 (200 mg, 0.31 mmol), cesium carbonate (201 mg, 0.62 mmol), and intermediate a7 (150 mg, 0.50 mmol) from the previous step were dissolved in 10 mL of anhydrous toluene. The catalyst bis(diphenylphosphine ether)palladium dichloride (44 mg, 0.06 mmol) was added. The reaction mixture was heated to 105 °C and reacted for 3 hours. The reaction was then stopped, filtered, and the solvent was removed by vacuum distillation. The crude product was separated by flash column chromatography (petroleum ether / ethyl acetate = 1 / 2) to give a pale yellow solid H33-3 (150 mg, 0.17 mmol), yield: 56%. LC-MS: [M+H] + =864.

[0437] Step 4: Under nitrogen protection, compound H33-3 (150 mg, 0.17 mmol) was dissolved in 3 mL of a mixed solution of trifluoroacetic acid and dichloromethane (v / v, 1 / 1). The reaction was carried out at room temperature for 1 hour. The reaction was then stopped, and saturated sodium bicarbonate solution was slowly added to the system to adjust the pH to approximately 8. The mixture was extracted with ethyl acetate and concentrated under reduced pressure. The residue was separated by flash column chromatography to obtain a yellow solid H33 (30 mg). LC-MS: [M+H] + =664.

[0438] 1 HNMR(400MHz,DMSO-d6)δ8.01(s,2H),7.84(s,1H),7.21(dd,J=8.3,5.4Hz,1H), 7.12(t,J=8.9Hz,1H),5.35–5.25(m,1H),5.13(dt,J=56.1,5.6Hz,1H),4.40(s,2 H),4.29–4.13(m,4H),3.52(s,2H),3.48–3.37(m,2H),2.81–2.66(m,3H),2.65– 2.61(m,2H),2.36(dd,J=15.3,8.2Hz,1H),2.10–1.80(m,9H),1.73–1.65(m,4H).

[0439] Example 22:

[0440]

[0441] Step 1: Using an ice bath, dissolve starting material H40-1 (600 mg, 2.54 mmol) and potassium tert-butoxide (404 mg, 3.6 mmol) in 5 mL of tetrahydrofuran and stir for 10 minutes. Pour the mixture into 10 mL of tetrahydrofuran containing intermediate a14 (882 mg, 1.8 mmol). Continue the reaction in an ice bath for 1 hour, then stop the reaction. Add 50 mL of ice water to the system, extract with ethyl acetate, dry to anhydrous sodium sulfate, concentrate, and separate the crude product by flash column chromatography to obtain compound H40-2 (500 mg, 0.71 mmol), yield 39%. LC-MS: [M+H] + =705.

[0442] Step 2: Under nitrogen protection, compound H40-2 (100 mg, 0.14 mmol), cesium carbonate (92 mg, 0.28 mmol), and intermediate a7 (80 mg, 0.20 mmol) from the previous step were dissolved in 5 mL of toluene. The catalyst bis(diphenylphosphine ether)palladium dichloride (10 mg, 0.014 mmol) was added, and the mixture was heated to 105 °C and reacted for 2 hours. The reaction was then stopped, and the mixture was filtered. The solvent was removed under reduced pressure, and the crude product was separated by flash column chromatography to obtain compound H40-3 (90 mg, 0.098 mmol), with a yield of 69%. LC-MS: [M+H] + =917.

[0443] Step 3: Dissolve compound H40-3 (90 mg) from the previous step in a mixed solution of trifluoroacetic acid and dichloromethane (v / v, 1 / 1). React at room temperature for 1 hour. Stop the reaction, slowly add saturated sodium bicarbonate solution to the system and adjust the pH to approximately 8. Extract with ethyl acetate and concentrate under reduced pressure. Separate the residue by TLC thin-layer chromatography to obtain a yellow solid H40 (10 mg). LC-MS: [M+H] + =597.

[0444] 1 H NMR(400MHz, DMSO-d6)δ8.14(s,2H),7.89(s,1H),7.32(dd,J=8.4,5.2Hz,1H),7.23–7.17(m,1H), 4.73(t,J=5.4Hz,1H),4.35–4.26(m,4H),3.64–3.35(m,7H),1.90–1.87(m,5H),1.73–1.60(m,5H).

[0445] Example 23:

[0446]

[0447] Step 1: Intermediate d2 (75 mg, 0.10 mmol) was dissolved in a mixed solution of trifluoroacetic acid and dichloromethane (3.0 mL, 1 / 1). The reaction was carried out in an ice bath for 1 hour, after which the reaction was stopped. The solvent was removed by vacuum distillation, and the crude product was separated by HPLC preparative chromatography to obtain compound M9 (30 mg). LC-MS: [M+H] + =555.

[0448] 1 H NMR(400MHz,DMSO-d6)δ8.18(s,2H),8.07(s,1H),7.28(dd,J=8.4,5.3Hz,1H ),7.22–7.15(m,1H),4.64(d,J=13.7Hz,1H),4.44(d,J=13.7Hz,1H),4.38(q ,J=7.1Hz,2H),4.18(d,J=27.8Hz,2H),4.03(d,J=13.4Hz,1H),3.84(d,J=13 .6Hz,1H),1.95(m,J=12.9Hz,2H),1.88–1.76(m,2H),1.34(m,J=7.1Hz,4H).

[0449] Example 24:

[0450]

[0451] Step 1: Under nitrogen protection, compound M1-1 (35 mg, 0.048 mmol), 4-dimethylaminopyridine (DMAP) (1.2 mg, 0.009 mmol), and cyclopropanol (5.0 mg, 0.072 mmol) were dissolved in 6 mL of dichloromethane. N,N'-dicyclohexylcarbodiimide (DCC) (45 mg, 0.096 mmol) was added, and the reaction was stopped at room temperature for 12 hours. 30 mL of ice water was added to the reaction solution, followed by extraction with dichloromethane. The solvent was removed under reduced pressure, and the crude product was separated by flash column chromatography (PE / EA = 3 / 1) to obtain compound M10-1 (38 mg). The yield was quantitative. LC-MS: [M+H] + =767.

[0452] Step 2: Dissolve compound M10-1 (38 mg) from the previous step in a mixed solution of trifluoroacetic acid and dichloromethane (3.0 mL, 1 / 1). React in an ice bath for 1 hour. Remove the solvent under reduced pressure. Separate the crude product by HPLC preparative chromatography to obtain compound M10 (8 mg). LC-MS: [M+H] + =567.

[0453] 1H NMR (400MHz, DMSO-d6) δ9.54(s,1H),8.16(s,2H),8.04(s,1H),7.29(dd,J=8.0,4.0Hz,1H),7.18(t,J=8.0Hz,1H),4.53(dd,J=76.0, 12Hz,2H),4.38–4.34(m,1H),3.93–3.71(m,4H),3.73(d,J=12Hz,1H),2.67(d,J=20Hz,1H),2.33(d,J=12Hz,1H),2.01–1.82(m,5H).

[0454] Following the synthetic route of compound M10, and using a similar skeletal structure, the following target molecule was synthesized.

[0455] * indicates a chiral molecule that has not undergone chiral resolution.

[0456]

[0457]

[0458] Example 25:

[0459]

[0460] Step 1: Using an ice bath, dissolve propionamide (1.0 g, 2.05 mmol) in 10 mL of anhydrous tetrahydrofuran, add NaH (123 mg, 3.07 mmol), and stir for 30 minutes. Then add intermediate a14 (976 mg, 2.0 mmol) to the reaction system. Heat to room temperature and react for 2 hours. Monitor the reaction for completeness using LC-MS. Add 50 mL of ice water to the reaction mixture, extract with ethyl acetate, dry to anhydrous sodium sulfate, concentrate, and separate the crude product by flash column chromatography to obtain compound M13-1 (663 mg, 1.23 mmol), yield: 61%. LC-MS: [M+H] + =542.

[0461] Step 2: Under nitrogen protection, compound M13-1 (300 mg, 0.55 mmol), cesium carbonate (360 mg, 1.11 mmol), and intermediate a7 (269 mg, 0.66 mmol) from the previous step were dissolved in 5 mL of toluene. The catalyst bis(diphenylphosphine ether)palladium dichloride (40 mg, 0.055 mmol) was added. The mixture was heated to 105 °C and reacted for 12 hours. The reaction was then stopped, and the mixture was filtered. The solvent was removed under reduced pressure. The crude product was separated by flash column chromatography to obtain compound M13-2 (85 mg, 0.11 mmol), yield: 21%. LC-MS: [M+H] + =754.

[0462] Step 3: Compound M13-2 (85 mg) from the previous step was dissolved in a mixed solution of trifluoroacetic acid and dichloromethane (3.0 mL, 1 / 1). The reaction was carried out in an ice bath for 1 hour. The solvent was removed by vacuum distillation. The crude product was separated by HPLC preparative chromatography to obtain compound M13 (15 mg). LC-MS: [M+H] + =554.

[0463] 1 H NMR (400MHz, DMSO) δ10.53(s,1H),8.14(s,2H),7.89(s,1H),7.26(dd,J=8.4,5.3Hz,1H),7.15(t,J=8.9Hz,1H),4.51(dd,J=79.5,13.4Hz,2 H), 4.10 (d, J = 21.7Hz, 2H), 3.95 (d, J = 13.1Hz, 1H), 3.76 (d, J = 13.3Hz, 1H), 2.54 (d, J = 7.4Hz, 2H), 2.05–1.76 (m, 5H), 1.05 (t, J = 7.5Hz, 3H).

[0464] Example 26:

[0465]

[0466] Step 1: Under nitrogen protection, 2-fluoro-5-methylaniline H44-1 (5.0 g, 39.96 mmol), potassium carbonate (13.8 g, 99.90 mmol), and potassium iodide (6.63 g, 39.96 mmol) were dissolved in 50 mL of N-methylpyrrolidone (NMP) and stirred for 5 minutes. 4-Methoxybenzyl chloride (12.83 g, 81.92 mmol) was slowly added to the mixture, and the mixture was heated to 65 °C and reacted for 1 hour. The reaction was then stopped. 200 mL of ice water was added to the system, and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by flash column chromatography to obtain compound H44-2 (14.0 g), yield: 95%. LC-MS: [M+H] + =366.

[0467] Step 2: Under nitrogen protection at -5℃, 2,2,6,6-tetramethylpiperidine (1.55 g, 11.0 mmol) was dissolved in 10 mL of anhydrous tetrahydrofuran, and n-butyllithium (4.38 mL, 2.5 M) was slowly added dropwise. After stirring for 10 minutes, the mixture was cooled to -60℃. Compound H44-2 (1.0 g, 2.74 mmol) from the previous step was added, and the mixture was stirred for 30 minutes. Triisopropyl borate (620 mg, 3.29 mmol) was added to the reaction solution, and the reaction was continued for another 30 minutes before being stopped. 50 mL of saturated ammonium chloride aqueous solution was slowly added to the reaction solution, and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by flash reversed column chromatography to obtain compound H44-3 (770 mg), yield: 68%. LC-MS: [M+H] + =:410.

[0468] Step 3: Under nitrogen protection, the compound H44-3 (209 mg, 0.51 mmol), cesium carbonate (499 mg, 1.53 mmol), and intermediate a14 (250 mg, 0.51 mmol) from the previous step were dissolved in 5 mL of 1,4-dioxane. Pd(dppf)Cl2 (39 mg, 0.052 mmol) was added as a catalyst, and the mixture was heated to 105 °C and reacted for 16 hours. The reaction was then stopped, and the mixture was filtered. The solvent was removed under reduced pressure, and the crude product was separated by flash column chromatography to obtain compound H44-4 (190 mg), yield: 48%. LC-MS: [M+H] + =774.

[0469] Step 4: Dissolve compound H44-4 (190 mg, 0.25 mmol) and N-iodosuccinimide (NIS) (62 mg, 0.28 mmol) in 2 mL of acetonitrile, add 2 drops of trifluoroacetic acid, and react at room temperature for 0.5 hours. Then stop the reaction. Pour the reaction solution into 20 mL of water, extract with ethyl acetate, dry to anhydrous sodium sulfate, concentrate, and separate the crude product by flash reversed column chromatography to obtain compound H44-5 (210 mg), yield: 95%. LC-MS: [M+H] + =900.

[0470] Step 5: Under nitrogen protection, compound H44-5 (140 mg, 0.16 mmol), methyl fluorosulfonyl difluoroacetate (308 mg, 1.6 mmol), and catalyst CuI (152 mg, 0.80 mmol) from the previous step were dissolved in 3 mL of DMF. The reaction was carried out at room temperature for 16 hours, and then the reaction was stopped. The reaction solution was poured into 30 mL of water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by flash reverse column chromatography to obtain compound H44-6 (70 mg), yield: 53%. LC-MS: [M+H] + =842.

[0471] Step 6: Under nitrogen protection, intermediate a18 (78 mg, 0.42 mmol) was dissolved in 4 mL of anhydrous tetrahydrofuran, and potassium tert-butoxide (95 mg, 0.84 mmol) was added. The mixture was stirred at room temperature for 30 minutes to obtain solution S0. In another reaction flask, compound H44-6 (350 mg, 0.42 mmol) was dissolved in 2 mL of anhydrous THF. Under ice bath conditions, the prepared solution S0 was slowly added, and stirring was continued for 1 hour. The reaction mixture was then stopped. The reaction solution was poured into 30 mL of ice water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by flash column chromatography to obtain a pale yellow solid H44-7 (300 mg). Yield: 71%, LC-MS: [M+H] + =1009.

[0472] Step 7: Dissolve compound H44-7 (300 mg, 0.30 mmol) from the previous step in 2 mL of toluene, add 2 mL of a mixed solution of trifluoroacetic acid and methanesulfonic acid (v / v, 1 / 1), heat to 50 °C and react for 16 hours, then stop the reaction. Remove the solvent under reduced pressure, and separate the residue by HPLC preparative chromatography to obtain a white solid H44 (20 mg). Yield: 9%, LC-MS: [M+H] + =669.

[0473] 1 H NMR (400MHz, DMSO-d6) δ7.89(s,1H),6.84(d,J=8.9Hz,1H),6.03(s,2H),5.11(dt,J=10.9,5.8Hz,1H),4.40(s,2 H),4.31–4.23(m,2H),3.64–3.46(m,5H),2.79–2.59(m,5H),2.38–2.32(m,4H),2.05–1.77(m,8H),1.67(s,4H).

[0474] Example 27:

[0475] The assay for the inhibition of KRAS G12D-mediated p-ERK by the compound (directly reflecting the cellular-level inhibitory effect of the test compound). Details are as follows:

[0476] AGS cells cultured in F-12K medium (Gibco, Cat. No. 30-2004) containing 10% fetal bovine serum and 1% penicillin-streptomycin were seeded into 384-well microplates and incubated overnight at 37°C with 5% carbon dioxide. 200 μL of different concentrations of the compound (dimethyl sulfoxide, final concentration 0.5%) were added to each well and incubated at 37°C for 3 hours. The cells were then fixed in 8% fixative (Solarbio, Cat. No. P1112) and washed once with phosphate-buffered saline (PBS). After washing, blocking solution (LI-COR, Cat. No. 927-40000) was added to each well and the cells were blocked at room temperature for 1 hour. After removing the blocking solution, add phospho-p44 / 42 MAPK (T202 / Y204) Rabbit mAb (CST, Cat. No. 97166S) and GAPDH (D4C6R) Mouse mAb (CST, Cat. No. 4370S) antibody working solutions to each well and incubate overnight at 4°C. Wash the microplate three times with PBS solution containing 0.1% Tween-80 (PBST), add IRDye 800CW Goat anti-Rabbit IgG (H+L) (LI-COR, Cat. No. 926-32211) and IRDye 680RD Goat anti-Mouse IgG (H+L) (LI-COR, Cat. No. 926-68070) antibody working solutions, and incubate the microplate at room temperature in the dark. After washing the microplate three times with PBST, the microplate was centrifuged at 1000 rpm for 1 minute, and the plate was scanned and the signal value was recorded using an Odyssey CLx (LI-COR) instrument.

[0477] IC 50 Calculation formula

[0478] Calculation of compound IC using nonlinear regression equations 50 Value: Y = Lower platform signal + (Upper platform signal - Lower platform signal) / (1 + 10^(LogIC)) 50 -X)*Hill slope)); X = logarithm of compound concentration.

[0479] Table 1: Inhibitory effect of compounds on p-ERK half-maximal concentration [MRTX1133 as positive control]

[0480] compound <![CDATA[p-ERK / IC 50 / nM]]> compound <![CDATA[p-ERK / IC 50 / nM]]> H1a 0.29 P1 54 H1b 4.6 P2 534 H2a 42 P3 450 H2b 1.1 P4 21 H3a 40 P5 169 H3b 0.33 P6 117 H4 2.6 P7 >1000 H5a >1000 P8 13 H5b 42 P9 0.88 H6 4.1 P10 1.9 H7 10 P11 2.8

[0481] H8 40 P12 259 H9 10 P13 18 H10a 98 P14 633 H10b 0.51 P15 158 H11 4.0 P16 94 H12a 112 P17 72 H12b 0.94 P18 7.9 H13 780 P19 3.6 H14 358 P20 0.77 H15 5.5 P21 13 H16 24 P22 3.0 H17 4.0 P23 13 H18 21 P24 0.42 H19a >1000 P25 5.2 H19b 7.5 P26 1.4 H20 3.2 P27 1.2 H21 51 P28 1.9 H22 23 P29 105 H23 25 M1 >1000 H24 1.9 M2 117 H25 5.5 M3 112 H26 >1000 M4 28 H27 3.9 M5 84 H28a 42 M6 29 H28b 1.6 M7 19 H29 >1000 M8 204 H30 >1000 M9 48 H31 >1000 M10 401 H32 >1000 M11 97 H33 37 M12 165 H34 69 M13 193 H35a 184 H35b 2.8 H36a 261 H36b 3.6 H37a 716 H37b 2.0 H38a 384 H38b 2.0 H39 31 H40 239 H41a 167 H41b 10 H42a 613 H42b 3.3 H43 37 H44 277 MRTX1133 1.0

[0482] MRTX1133 structure:

[0483]

[0484] Example 28: Inhibitory activity of the compound against GTP-KRAS

[0485] The analyte was serially diluted 4-fold. Using ECHO (Labcyte), 0.1 μL of different concentrations of the analyte was transferred to each well of a 384-well microplate. 5 μL of diluted Tag2-KRAS G12D>P or Tag2-KRASWT>P was added to each well sequentially, and the plates were centrifuged at 1000 RPM for 1 minute. Then, 5 μL of diluted Tag1-cRAF was added to each well, and the plates were centrifuged at 1000 RPM for 1 minute and incubated at 25°C for 15 minutes. Finally, 10 μL of a mixture of anti-Tag1-Tb3 and anti-Tag2-XL665 was added to each well, and the plates were centrifuged at 1000 RPM for 1 minute and incubated at 4°C for 3 hours. The plates were then scanned and the signal values ​​were recorded using Envision at 665 / 615 nm.

[0486] In the above analysis, all KRAS-G12D detection reagents were obtained from the commercially available kit KRAS-G12D / cRAFBINDING ASSAY KITS (Cisbio, Cat. No. 63ADK000CB21PEG); for KRAS-WT detection, GTP was purchased from Sigma (Cat. No. V900868), GST-cRAF was prepared by Beijing Kanglong Chemical (Cat. No. 20190718), MAb AntiGST-Tb cryptate was purchased from Cisbio (Cat. No. 61GSTTLA), and other key reagents were obtained from the commercially available kit KRAS-WT / SOS1 BINDING ASSAY KITS (Cisbio, Cat. No. 63ADK000CB15PEH).

[0487] The calculation results are analyzed based on the following formula:

[0488] Relative ratio (RR) = (Ratio 665 / 615 -Ratio 背景 )

[0489] Inhibition percentage = [1-(RR)] 化合物 -RR 阳性对照孔 (average) / (RR) 阴性对照孔 Average - RR 阳性对照孔 [Average value] × 100

[0490] IC 50 Calculate: Y = Lower platform signal + (Upper platform signal - Lower platform signal) / (1 + 10^(LogIC)) 50-X)×Hill slope). X: Logarithmic value of compound concentration; Y: Inhibition percentage.

[0491] Table 2. Inhibitory effects of compounds on GTP-KRAS

[0492] compound <![CDATA[KRAS G12D-CRAF / IC 50 / nM]]> H1a 5.1 H2b 9.2 H4 7.4 H10b 8.5 H19b 10 P4 16 P20 11 MRTX1133 11

[0493] These results indicate that the molecule of the present invention has an excellent inhibitory effect on the activation of KRAS G12D protein.

[0494] Example 29:

[0495] The compound exhibits a 3D antiproliferative effect against the KRAS G12D-mutant pancreatic cancer AsPC-1 cell line. Details are as follows:

[0496] Cell culture: AsPC-1 pancreatic cancer cells were cultured in T75 culture flasks (Corning, catalog number 430641) in RPMI 1640 medium (Hyclone, catalog number SH3080901B) containing 10% fetal bovine serum (Ausgenex, catalog number FBS500-S) and 1% penicillin / streptomycin (Gibco, catalog number 15140-122).

[0497] Experimental Procedure: The diluted test compound was added to a 384-well low-adsorption cell culture plate (Labcyte, PP-0200) using a nanoliter pipetting system (LABCYTE, catalog number Echo550). After cell seeding, the plate was incubated at 37°C in a 5% CO2 incubator. The test compound (1 μM as the starting concentration, 3-fold dilution, for a total of 10 concentrations) was co-incubated with the cells for 7 days. Then, the solution was added to each well. The 3D reagent (Promega, catalog number G9683) was used, and the luminescence value was read using an Envision multi-functional microplate reader (Perkin Elmer, catalog number Envision 2104). The light signal was directly proportional to the amount of ATP in the system, and the ATP content directly represented the number of viable cells in the system. Finally, the IC50 of the compound was obtained using a nonlinear fitting formula with XLFIT software. 50 (Half-maximal inhibitory concentration).

[0498] Inhibition rate (%) = 100 × (Negative control average value - Compound reading) / (Negative control average value - Positive control average value)

[0499] Negative control: DMSO. Positive control: Culture medium.

[0500] Table 3: Antiproliferative effects of compounds at half-maximal effective concentrations on AsPC-1 cells

[0501] compound <![CDATA[IC 50 / nM]]> H1a 4.2 H1b 49 H2b 23 P4 851 P11 50 P20 47

[0502] These results indicate that the molecules of this invention have a good anti-proliferative effect on KRAS G12D-mutated tumor cell lines.

[0503] Example 30:

[0504] The compound exhibits a 3D antiproliferative effect against the KRAS G12D-mutant pancreatic cancer PANC-1 cell line. Details are as follows:

[0505] Cell culture: PANC-1 pancreatic cancer cells were cultured in T75 culture flasks (Corning, catalog number 430641) in DMEM medium (GIBCO) containing 10% fetal bovine serum (Ausgenex, catalog number FBS500-S) and 1% penicillin / streptomycin (Gibco, catalog number 15140-122).

[0506] Experimental Procedure: The diluted test compound was added to a 384-well low-adsorption cell culture plate (Labcyte, PP-0200) using a nanoliter pipetting system (LABCYTE, catalog number Echo550). After cell seeding, the plate was incubated at 37°C in a 5% CO2 incubator. The test compound (1 μM as the starting concentration, 3-fold dilution, for a total of 10 concentrations) was co-incubated with the cells for 7 days. Then, the solution was added to each well. The 3D reagent (Promega, catalog number G9683) was used, and the luminescence value was read using an Envision multi-functional microplate reader (Perkin Elmer, catalog number Envision 2104). The light signal was directly proportional to the amount of ATP in the system, and the ATP content directly represented the number of viable cells in the system. Finally, the IC50 of the compound was obtained using a nonlinear fitting formula with XLFIT software. 50 (Half-maximal inhibitory concentration).

[0507] Inhibition rate (%) = 100 × (Negative control average value - Compound reading) / (Negative control average value - Positive control average value)

[0508] Negative control: DMSO. Positive control: Culture medium.

[0509] Table 4: Antiproliferative effects of compounds at half-maximal effective concentrations on PANC-1 cells

[0510] compound <![CDATA[IC 50 / nM]]> H2b 1.3 H3b 1.0 H10b 1.8 H12b 2.4 H19b 7.7 P20 3.0 MRTX1133 3.6

[0511] These results indicate that the molecules of this invention have a good anti-proliferative effect on KRAS G12D-mutated tumor cell lines.

[0512] Example 31:

[0513] Liver microsomal stability assay of the compound. Details are as follows:

[0514] The liver microsomal stability of the compounds of this invention was studied. The test compounds were co-incubated with liver microsomes of different species with or without the addition of NADPH. The final concentrations of the test compounds, NADPH, and liver microsomes in the test system were 1 μM, 1 mM, and 0.5 mg / ml, respectively. The concentrations of the compounds in the supernatant at different time points within 60 minutes were measured, and pharmacokinetic parameters (e.g., clearance rate Clint) were calculated.

[0515] This result indicates that the molecules of the present invention have good metabolic stability (especially in the human body).

[0516] Some molecules (such as H1b, H10b, H24, H25, H35b-H38b, P20, P24, etc.) have lower clearance rates and slower metabolism in the human body compared to the control MRTX1133.

[0517] compound Human Clint (mL / min / kg) MouseClint / (mL / min / kg) H1a 27 451 H1b 9.7 456 H2b 40.4 156 H3b 44.9 229 H4 33.7 211.7 H5b 32.9 244 H10b 14.2 167 H11 39.6 298 H12b 24.1 182 H19b 32.9 177 H24 19.8 166 H25 10.3 ND H35b 17.8 143 H36b 12.7 102 H37b 14.3 110 H38b 20.0 152 P4 40.8 595 P11 29.7 547 P20 16.2 334 P24 20.0 403 MRTX1133 28.7 260

[0518] ND = Untested

[0519] Example 32:

[0520] In vivo efficacy study of BALB / c in nude mice. Details are as follows:

[0521] KRAS G12D-mutated colorectal cancer tumor cells (GP2D) were cultured and subcutaneously inoculated into 6-8 week old female BALB / c nude mice (approximately 20g in weight). Mice were cultured in an SPF-grade experimental environment, and all mice had free access to a commercially certified standard diet. The tumors were cultured when the average tumor volume reached 150 mm. 3 Around 10 days post-treatment, the test compound was administered intraperitoneally (ip) daily. The dosage was the same as the control group (10% Captisol in 50mM citrate buffer, pH 5.0). The administered group received 10 mg / kg twice daily. Tumor volume was measured three times a week using calipers, and animals were weighed daily. After 10 days of continuous administration, the inhibition rate (TGI / 100%) was calculated based on the final tumor volume. The volume calculation formula was: V = 1 / 2a*b 2 , where a represents the long diameter of the tumor and b represents the short diameter of the tumor.

[0522] test drug Dosage TGI Blank group 0 0％ MRTX1133 10mg / kg, BID 186％ P20 10mg / kg, BID 187％

[0523] The results indicate that the molecule of the present invention has good in vivo efficacy, can inhibit the growth of KRAS G12D mutant tumors, and its effect is better than that of MRTX1133.

[0524] Example 33:

[0525] Animal in vivo safety experiments of the compounds of this invention are detailed below:

[0526] Select qualified healthy ICR mice (6-8 weeks old, weighing 18-20g), with 3 mice in each group, and administer a single intravenous dose. A single intravenous administration trial was conducted first, starting with a dose of 2mg / kg. If no deaths were observed, the dose was increased; if deaths occurred, the dose was stopped.

[0527] The intravenous administration solution of MRTX1133 and the intravenous administration solvent of compound P20 are: DMSO / Tween 80 / Solutol / physiological saline (volume ratio of the four is 5 / 3 / 10 / 82). After being fully dissolved by vortex sonication, the solution is administered.

[0528] compound Administration method Dosage Mortality rate / 3 birds MRTX1133 IV 2mpk 0 / 3 MRTX1133 IV 4mpk 1 / 3 P20 IV 2mpk 0 / 3 P20 IV 4mpk 0 / 3 P20 IV 8mpk 0 / 3 P20 IV 16mpk 2 / 3

[0529] Healthy, qualified ICR mice (6-8 weeks old, weighing 18-20g) were selected, with 3 mice per group, and each mouse was given a single intravenous infusion. The maximum intravenous dose was started; if no deaths were observed, the dose was increased; if deaths occurred, the dose was stopped. The infusion solution for MRTX1133 and the infusion solvent for compound P20 were: DMSO / Tween 80 / Solutol / physiological saline (volume ratio of 5 / 3 / 10 / 82). The solution was vortexed and sonicated to ensure complete dissolution before administration.

[0530] compound Administration method Dosage Mortality rate / 3 birds MRTX1133 Inf input 4mpk 0 / 3 MRTX1133 Inf input 8mpk 0 / 3 MRTX1133 Inf input 16mpk 0 / 3 MRTX1133 Inf input 32mpk 2 / 3 P20 Inf input 16mpk 0 / 3 P20 Inf input 32mpk 0 / 3 P20 Inf input 96mpk 0 / 3 P20 Inf input 150mpk 0 / 3

[0531] The results indicate that the molecule of the present invention has good in vivo safety, and its safety is far superior to that of MRTX1133, whether administered intravenously or by infusion.

[0532] Example 34:

[0533] In vivo pharmacokinetic studies of the compound in mice. Details are as follows:

[0534] CD1 female mice were used as test animals, and the drugs were administered orally / intravenously (the oral dose was 20 mg / kg, and the solvent was DMSO-Solutol-H2O).

[0535] Experimental protocol: Three mice were administered the oral medication. Plasma samples were collected before (0 h) and after (0.25, 0.5, 1, 2, 4, 8, 24 h) administration. The plasma concentrations of mice after oral administration were determined by LC / MS / MS. The collected data were calculated using AB Sciex QTRAP 6500 software. The experimental results are as follows:

[0536]

[0537]

[0538] * = 50mg / kg

[0539] These results indicate that the molecules of this invention have good in vivo absorption in mice, and some molecules have very high in vivo exposure levels in mice, which are much higher than MRTX1133. Clinically, oral administration may be possible.

Claims

1. A compound, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of:

2. A pharmaceutical composition comprising a compound of claim 1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

3. The pharmaceutical composition of claim 2, further comprising another therapeutic agent.

4. Use of a compound of claim 1, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment and / or prevention of a KRAS G12D mutant protein-mediated disease; wherein the KRAS G12D mutant protein-mediated disease is selected from the group consisting of pancreatic cancer, colorectal cancer, and lung cancer.

5. The use of claim 4, the lung cancer is selected from the group consisting of non-small cell lung cancer (NSCLC) and small cell lung cancer.

Images