Wnt pathway inhibitor compounds

Abstract

The present invention provides a compound of formula I having excellent Wnt pathway inhibitory activity or a pharmaceutically acceptable salt, isotope derivative, stereoisomer thereof. The present invention also provides a method for preparing the compound and its use in preventing and / or treating cancer, tumors, inflammatory diseases, autoimmune diseases or immune-mediated diseases.

Description

[0001] This application claims priority to a Chinese patent application filed with the State Intellectual Property Office of China on January 30, 2022, with application number 202210115506.1 and invention name “Wnt pathway inhibitor compounds”, all contents of which are incorporated by reference into this application. Technical Field

[0002] The present invention relates to a heterocyclic compound, in particular to a highly active Wnt pathway inhibitor and application thereof. Background Art

[0003] The Wnt / β-catenin signal transduction pathway is a conserved pathway in biological evolution. In normal somatic cells, β-catenin only functions as a cytoskeletal protein to form a complex with E-cadherin at the cell membrane to maintain the adhesion of homotypic cells and prevent cell movement. When the Wnt signaling pathway is not activated, β-catenin in the cytoplasm is phosphorylated and forms a β-catenin degradation complex with APC, Axin, and GSK3β, thereby initiating the ubiquitin system to degrade β-catenin via the proteasome pathway, maintaining β-catenin in the cytoplasm at a low level. When cells are stimulated by Wnt signals, Wnt proteins bind to the specific receptor Frizzled protein on the cell membrane. The activated Frizzled receptor recruits intracellular Dishevelled protein, inhibits the degradation activity of the β-catenin degradation complex formed by GSK3β and other proteins, and stabilizes the free β-catenin protein in the cytoplasm. After β-catenin stably accumulated in the cytoplasm enters the cell nucleus, it binds to the LEF / TCF transcription factor family and initiates the transcription of downstream target genes (such as c-myc, c-jun, Cyclin D1, etc.). Overactivation of the Wnt / β-catenin signaling pathway is closely related to the occurrence of various cancers (including colon cancer, gastric cancer, breast cancer, etc.). For example, abnormal activation of the Wnt classical signaling pathway and nuclear accumulation of β-catenin protein are widely present in colorectal cancer, and the proliferation of cancers such as colon cancer can be inhibited by inhibiting the activity of the Wnt signaling pathway. APC mutations exist in more than 85% of colorectal cancers. The mutated APC blocks the phosphorylation and degradation of β-catenin, inducing the occurrence of colorectal cancer. In addition, Axin mutations and β-catenin mutations can also cause intracellular aggregation of β-catenin and activate the Wnt / β-catenin pathway.

[0004] Although it is known that inhibiting the Wnt signaling pathway can effectively prevent and / or treat cancer, tumors, inflammatory diseases, autoimmune diseases and immune-mediated diseases, there is currently a lack of satisfactory and effective Wnt pathway inhibitor compounds in the prior art. Therefore, the study of effective Wnt pathway inhibitor compounds is a need in the prior art. Summary of the invention

[0005] One aspect of the present invention provides a class of compounds having a structure of Formula I or a pharmaceutically acceptable salt, isotopic derivative, or stereoisomer thereof:

[0006]

[0007] Among them, X 1 , X 2 Both are N, or one of them is N and the other is CH;

[0008] R 1 , R 2 Each independently represents hydrogen, halogen, (C 1 -C 6 ) alkyl, halogenated (C 1 -C 6 )alkyl;

[0009] R 3 represents hydrogen, halogen, (C 1 -C 6 ) alkyl, halogenated (C 1 -C 6 )alkyl;

[0010] R 4 represents hydrogen, halogen, (C 1 -C 6 ) alkyl, halogenated (C 1 -C 6 ) alkyl, or R 4 With R 2 Forming 4-8 membered rings;

[0011] R 5 Each independently represents hydrogen, halogen, (C 1 -C 6 ) alkyl, halogenated (C 1 -C 6 ) alkyl, (C 3 -C 8 ) cycloalkyl, halo(C 3 -C 8 ) cycloalkyl, or two R 5 Forming a 3-5 membered ring; m is 0, 1, 2 or 3;

[0012] R6 Each independently represents hydrogen, halogen, -CN, (C 1 -C 6 ) alkyl, halogenated (C 1 -C 6 )alkyl;

[0013] R 7 represents hydrogen, halogen, (C 1 -C 6 ) alkyl, halogenated (C 1 -C 6 ) alkyl, (C 3 -C 8 ) cycloalkyl, halo(C 3 -C 8 )cycloalkyl, -OR a 、-halogenated OR a 、-SR a ,-halogenated SR a ;

[0014] R 8 represents hydrogen, -(C 1 -C 6 )alkyl, -halogenated (C 1 -C 6 )alkyl;

[0015] R 9 Represents halogen, (C 1 -C 6 ) alkyl, halogenated (C 1 -C 6 ) alkyl, (C 3 -C 8 ) cycloalkyl, halo(C 3 -C 8 )cycloalkyl, -(C 1 -C 6 )alkyleneCN, -halogenated(C 1 -C 6 )alkyleneCN, -(C 3 -C 8 )cycloalkyleneCN, -halogenated(C 3 -C 8 ) cycloalkylene CN, -NR a R a '、-(C 1 -C 6 )Alkylene NR a R a ', -halogenated (C 1 -C 6 )Alkylene NR a R a ', where R a, R a 'Can form a 4-8 membered ring with the N to which it is connected;

[0016] R a , R a ' each independently represents hydrogen, (C 1 -C 6 ) alkyl, halogenated (C 1 -C 6 ) alkyl, (C 3 -C 8 )cycloalkyl or halogenated (C 3 -C 8 )cycloalkyl.

[0017] In one embodiment of the present invention, R 4 Selected from (C 1 -C 6 ) alkyl, halogenated (C 1 -C 6 )alkyl.

[0018] In one embodiment of the present invention, R 5 is selected from hydrogen, halogen, (C 1 -C 6 ) alkyl, halogenated (C 1 -C 6 ) alkyl. In one embodiment of the present invention, R 6 For hydrogen.

[0019] In one embodiment of the present invention, X 1 , X 2 At the same time, N, R 9 Selected from (C 1 -C 6 ) alkyl, halogenated (C 1 -C 6 ) alkyl, (C 3 -C 8 ) cycloalkyl, halo(C 3 -C 8 )cycloalkyl.

[0020] In one embodiment of the present invention, when X 1 , X 2 When both are N, R 7 Not halogen.

[0021] In one embodiment of the present invention, R 7 Selected from (C 1 -C 6 ) alkyl, halogenated (C 1 -C 6 )alkyl.

[0022] In one embodiment of the present invention, R 5 It is a halogen.

[0023] In one embodiment of the present invention, R 5 For fluorine.

[0024] In one embodiment of the present invention, when X 1 , X 2 When both are N, R 9 Represents halogen, (C 1 -C 6 ) alkyl, halogenated (C 1 -C 6 ) alkyl, (C 3 -C 8 ) cycloalkyl, halo(C 3 -C 8 )cycloalkyl, -(C 1 -C 6 )alkyleneCN, -halogenated(C 1 -C 6 )alkyleneCN, -(C 3 -C 8 )cycloalkyleneCN, -halogenated(C 3 -C 8 ) cycloalkylene CN, -NR a R a '、-(C 1 -C 6 )Alkylene NR a R a ', -halogenated (C 1 -C 6 )Alkylene NR a R a ', where R a , R a ' can form a 4-8 membered ring with the N to which it is connected.

[0025] In one embodiment of the present invention, when X 1 , X 2 When one of them is N and the other is CH, R 9 Represents halogen, -(C 1 -C 6 )alkyleneCN, -halogenated(C 1 -C 6 )alkyleneCN, -(C 3 -C 8 )cycloalkyleneCN, -halogenated(C 3 -C 8 ) cycloalkylene CN, -NR a R a '、-(C1 -C 6 )Alkylene NR a R a ', -halogenated (C 1 -C 6 )Alkylene NR a R a ', where R a , R a ' can form a 4-8 membered ring with the N to which it is connected.

[0026] In one embodiment of the present invention, when X 1 , X 2 When one of them is N and the other is CH, R 9 Indicates (C 1 -C 6 ) alkyl, halogenated (C 1 -C 6 ) alkyl, (C 3 -C 8 ) cycloalkyl, halo(C 3 -C 8 )cycloalkyl.

[0027] In another aspect of the present invention, there is also provided a compound having the following structure or a pharmaceutically acceptable salt, isotopic derivative, stereoisomer thereof, wherein the compound has the following structure:

[0028]

[0029]

[0030]

[0031]

[0032]

[0033]

[0034] In another aspect of the present invention, a pharmaceutical composition is provided, comprising any of the aforementioned compounds or pharmaceutically acceptable salts, isotopic derivatives, stereoisomers thereof, and an optional pharmaceutically acceptable carrier.

[0035] In another aspect of the present invention, there is also provided the use of the aforementioned compound or its pharmaceutically acceptable salt, isotope derivative, stereoisomer or the aforementioned pharmaceutical composition in the preparation of a medicament for preventing and / or treating cancer, tumor, inflammatory disease, autoimmune disease or immune-mediated disease.

[0036] It is specifically noted that, herein, when referring to a "compound" of the structure of Formula I, stereoisomers, diastereomers, enantiomers, racemic mixtures and isotopic derivatives thereof are generally also encompassed.

[0037] It is well known to those skilled in the art that a salt, solvate, or hydrate of a compound is an alternative form of the compound, and they can all be converted into the compound under certain conditions. Therefore, it should be noted that when referring to a compound of Formula I herein, it generally also includes its pharmaceutically acceptable salts, and further includes its solvates and hydrates.

[0038] Similarly, when a compound is referred to herein, prodrugs, metabolites, and N-oxides thereof are also generally included.

[0039] Pharmaceutically acceptable salts of the present invention may be formed using, for example, the following inorganic or organic acids: "pharmaceutically acceptable salts" refer to salts that are suitable for contact with tissues of humans and lower animals, without undue toxicity, irritation, allergic reactions, etc., within the scope of reasonable medical judgment, and are considered to be a reasonable benefit / risk ratio. The salts may be prepared in situ during the final isolation and purification of the compounds of the present invention, or separately by reacting the free base or free acid with a suitable reagent, as outlined below. For example, the free base function may be reacted with a suitable acid. Examples of pharmaceutically acceptable inorganic acid addition salts are salts formed of an amino group with an inorganic acid (e.g., hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid) or an organic acid (e.g., acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid), or by using other methods known in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, sodium alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate and the like. Representative alkali metal or alkaline earth metal salts include salts of sodium, lithium, potassium, calcium, magnesium, etc. Other pharmaceutically acceptable salts include (where appropriate) non-toxic ammonium salts, quaternary ammonium salts, and amine cations formed with counterions, for example, halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates, and aryl sulfonates.

[0040] The pharmaceutically acceptable salts of the present invention can be prepared by conventional methods, for example, by dissolving the compound of the present invention in a water-miscible organic solvent (e.g., acetone, methanol, ethanol and acetonitrile), adding an excess of an organic acid or an aqueous inorganic acid solution thereto to precipitate the salt from the resulting mixture, removing the solvent and the remaining free acid therefrom, and then isolating the precipitated salt.

[0041] The precursors or metabolites of the present invention may be precursors or metabolites known in the art, as long as the precursors or metabolites are converted into compounds through in vivo metabolism. For example, "prodrugs" refer to those prodrugs of the compounds of the present invention, which are suitable for contact with human and lower animal tissues within the scope of reasonable medical judgment, without undue toxicity, irritation, allergic reactions, etc., and are considered to have a reasonable benefit / risk ratio and are effective for their intended use. The term "prodrug" refers to a compound that is rapidly converted in vivo to produce the parent compound of the above formula, for example, by in vivo metabolism, or N-demethylation of the compounds of the present invention.

[0042] "Solvate" as used herein means a physical association of a compound of the invention with one or more solvent molecules (whether organic or inorganic). The physical association includes hydrogen bonding. In some cases, for example, when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid, the solvate will be able to be separated. The solvent molecules in the solvate may exist in a regular arrangement and / or a disordered arrangement. The solvate may contain stoichiometric or non-stoichiometric amounts of solvent molecules. "Solvate" encompasses solution phases and separable solvates. Exemplary solvates include, but are not limited to, hydrates, ethanolates, methanolates, and isopropanolates. Solvation methods are well known in the art.

[0043] The "stereoisomerism" described in the present invention is divided into conformational isomerism and configurational isomerism. Configurational isomerism can also be divided into cis-trans isomerism and optical isomerism (i.e. optical isomerism). Conformational isomerism refers to a stereoisomerism phenomenon in which the atoms or atomic groups of molecules have different arrangements in space due to the rotation or distortion of carbon and carbon single bonds in organic molecules with a certain configuration. Common structures include the structures of alkane and cycloalkane compounds, such as the chair conformation and boat conformation that appear in the structure of cyclohexane. "Stereoisomer" refers to when the compounds of the present invention contain one or more asymmetric centers, and thus can be used as racemates and racemic mixtures, single enantiomers, diastereomeric mixtures and single diastereoisomers. The compounds of the present invention have asymmetric centers, and each asymmetric center will produce two optical isomers. The scope of the present invention includes all possible optical isomers and diastereomeric mixtures and pure or partially pure compounds. The compounds described in the present invention can exist in the form of tautomers, which have different hydrogen connection points due to one or more double bond displacements. For example, a ketone and its enol form are keto-enol tautomers. Each tautomer and mixtures thereof are included in the compounds of the present invention. Enantiomers, diastereomers, racemates, mesomorphs, cis-trans isomers, tautomers, geometric isomers, epimers and mixtures thereof of all compounds of formula (I) are included in the scope of the present invention.

[0044] The "isotopic derivative" of the present invention refers to a molecule in which the compound is isotopically labeled. The isotopes commonly used as isotopic labels are: hydrogen isotopes, 2 H and 3 H; Carbon isotope: 11 C, 13 C and 14 C; Chlorine isotope: 35 Cl and 37 Cl; Fluorine isotope: 18 F; Iodine isotope: 123 I and 125 I; Nitrogen isotopes: 13 N and 15 N; oxygen isotope: 15 Oh, 17 O and 18 O and sulfur isotopes 35 S. These isotope-labeled compounds can be used to study the distribution of drug molecules in tissues. 3 H and carbon 13 C, because they are easy to label and detect, they are more widely used. Some heavy isotopes, such as deuterium ( 2H), the substitution can enhance metabolic stability, prolong half-life and thus achieve the purpose of reducing dosage and provide therapeutic advantages. Isotope-labeled compounds are generally synthesized from labeled starting materials using known synthesis techniques in the same way as non-isotope-labeled compounds.

[0045] The present invention also provides the use of the compound of the present invention in preparing a medicament for preventing and / or treating cancer, tumor, inflammatory disease, autoimmune disease or immune-mediated disease.

[0046] In addition, the present invention provides a pharmaceutical composition for preventing and / or treating cancer, tumors, inflammatory diseases, autoimmune diseases, neurodegenerative diseases, attention-related diseases or immune-mediated diseases, which comprises the compound of the present invention as an active ingredient. The pharmaceutical composition may optionally comprise a pharmaceutically acceptable carrier.

[0047] In addition, the present invention provides a method for preventing and / or treating cancer, tumors, inflammatory diseases, autoimmune diseases, neurodegenerative diseases, attention-related diseases or immune-mediated diseases, which comprises administering a compound of formula I of the present invention or a pharmaceutically acceptable salt, isotope derivative, stereoisomer or pharmaceutical composition of the present invention to a mammal in need thereof.

[0048] Representative examples of inflammatory diseases, autoimmune diseases, and immune-mediated diseases may include, but are not limited to, arthritis, rheumatoid arthritis, spondyloarthritis, gouty arthritis, osteoarthritis, juvenile arthritis, other arthritic conditions, lupus, systemic lupus erythematosus (SLE), skin-related diseases, psoriasis, eczema, dermatitis, atopic dermatitis, pain, lung disease, lung inflammation, adult respiratory distress syndrome (ARDS), pulmonary sarcoidosis, chronic inflammatory lung disease, chronic obstructive pulmonary disease (COPD), cardiovascular disease, atherosclerosis, myocardial infarction, congestive heart failure, myocardial ischemia-reperfusion injury, inflammatory bowel disease, Crohn's disease, ulcerative colitis, irritable bowel syndrome, asthma, Sjögren's syndrome, autoimmune thyroid disease, disease, urticaria (urticaria), multiple sclerosis, scleroderma, organ transplant rejection, xenotransplantation, idiopathic thrombocytopenic purpura (ITP), Parkinson's disease, Alzheimer's disease, diabetes-related diseases, inflammation, pelvic inflammatory disease, allergic rhinitis, allergic bronchitis, allergic sinusitis, leukemia, lymphoma, B-cell lymphoma, T-cell lymphoma, myeloma, acute lymphoid leukemia (ALL), chronic lymphoid leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), hairy cell leukemia, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, myelodysplastic syndrome (MDS), myeloproliferative neoplasms (MPN), diffuse large B-cell lymphoma and follicular lymphoma.

[0049] Representative examples of cancer or tumors may include, but are not limited to, skin cancer, bladder cancer, ovarian cancer, breast cancer, stomach cancer, pancreatic cancer, prostate cancer, colon cancer, lung cancer, bone cancer, brain cancer, neuroblastoma, rectal cancer, colon cancer, familial adenomatous polyposis carcinoma, hereditary nonpolyposis colorectal cancer, esophageal cancer, lip cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, gastric cancer, adenocarcinoma, medullary thyroid cancer, papillary thyroid cancer, kidney cancer, renal parenchymal cancer, ovarian cancer, cervical cancer, uterine corpus cancer, endometrial cancer, choriocarcinoma, pancreatic cancer, prostate cancer, testicular cancer, urinary cancer, melanoma, brain tumors such as glioblastoma, astrocytoma, meningioma, medulloblastoma and peripheral Neuroectodermal tumor, Hodgkin lymphoma, non-Hodgkin lymphoma, Burkitt lymphoma, acute lymphoid leukemia (ALL), chronic lymphoid leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), adult T-cell leukemia lymphoma, diffuse large B-cell lymphoma (DLBCL), hepatocellular carcinoma, gallbladder cancer, bronchial carcinoma, small cell lung cancer, non-small cell lung cancer, multiple myeloma, basal cell tumor, teratoma, retinoblastoma, choroidal melanoma, seminoma, rhabdomyosarcoma, craniopharyngioma, osteosarcoma, chondrosarcoma, myosarcoma, liposarcoma, fibrosarcoma, Ewing sarcoma, or plasmacytoma.

[0050] When the compound of the present invention or a pharmaceutically acceptable salt thereof is administered in combination with another anticancer agent or immune checkpoint inhibitor for treating cancer or tumor, the compound of the present invention or a pharmaceutically acceptable salt thereof may provide an enhanced anticancer effect.

[0051] Representative examples of anticancer agents for treating cancer or tumors may include, but are not limited to, cell signaling inhibitors, chlorambucil, guanfalan, cyclophosphamide, ifosfamide, busulfan, carmustine, lomustine, streptozotocin, cisplatin, carboplatin, oxaliplatin, dacarbazine, temozolomide, procarbazine, methotrexate, fluorouracil, cytarabine, gemcitabine, mercaptopurine, fludarabine, vinblastine, vincristine, vinorelbine, paclitaxel, docetaxel, topotecan, irinotecan, etoposide, trabectedin, dactinomycin, doxorubicin, epirubicin, daunorubicin, mitoxantrone, bleomycin, mitomycin C, ixapric acid, thiocyanate, chloramphenicol ... lon, tamoxifen, flutamide, gonadorelin analogs, megestrol acetate, prednisone, dexamethasone, methylprednisolone, thalidomide, interferon alfa, leucovorin, sirolimus, temsirolimus, everolimus, afatinib, alisertib, amuvatinib, apatinib, axitinib, bortezomib, bosutinib, brivanib, cabozantinib, cediranib, crenolanib, crizotinib, dabrafenib, dacomitinib, danusetinib, dasatinib, dovitinib, erlotinib, foretinib, ganetespib, gefitinib, ibrutinib, icotinib, imatinib tinib, iniparib, lapatinib, lenvatinib, linifanib, linsitinib, masitinib, momelotinib, motesanib, neratinib, nilotinib, niraparib, oprozomib, olaparib, pazopanib, pictilisib, ponatinib, quizartinib, regorafenib, rigosertib, rucaparib, ruxolitinib, saricatinib, saridegib, sorafenib, sunitinib, telatinib, tivantinib, Tivozanib, tofacitinib, traguantinib, vandetanib, veliparib, vemurafenib, vismodegib, volasertib, alemtuzumab, bevacizumab, berentuzumab vedotin, catumaxomab, cetuximab, denosumab, gemtuzumab, ipilimumab, nimotuzumab, ofatumumab, panitumumab, rituximab, tositumomab, trastuzumab, PI3K inhibitor, CSF1R inhibitor, A2A and / or A2B receptor antagonist, IDO inhibitor, anti-PD-1 antibody, anti-PD-L1 antibody, LAG3 antibody, TIM-3 antibody and anti-CTLA-4 antibody, or any combination thereof.

[0052] When the compounds of the present invention or their pharmaceutically acceptable salts are administered in combination with another therapeutic agent for treating inflammatory diseases, autoimmune diseases and immune-mediated diseases, the compounds of the present invention or their pharmaceutically acceptable salts may provide enhanced therapeutic effects.

[0053] Representative examples of therapeutic agents for treating inflammatory diseases, autoimmune diseases and immune-mediated diseases may include, but are not limited to, steroidal drugs (e.g., prednisone, hydroprednisolone, methylhydroprednisolone, cortisone, hydroxycortisone, betamethasone, dexamethasone, etc.), methotrexate, leflunomide, anti-TNFα agents (e.g., etanercept, infliximab, adalimumab, etc.), calcineurin inhibitors (e.g., tacrolimus, piguanlimus, etc.) and antihistamines (e.g., diphenhydramine, hydroxyzine, loratadine, ebastine, ketotifen, cetirizine, levocetirizine, fexofenadine, etc.), and at least one or more therapeutic agents selected therefrom may be contained in the pharmaceutical composition of the present invention.

[0054] The compound of the present invention or its pharmaceutically acceptable salt can be used as an active ingredient. The dosage of the active ingredient can be adjusted according to multiple relevant factors (such as the condition of the subject to be treated, the type of disease and severity, the rate of administration and the doctor's opinion). In some cases, an amount less than the above dosage may be suitable. If no harmful side effects are caused, an amount greater than the above dosage can be used and the amount can be administered in divided doses every day.

[0055] In addition, the present invention also provides a method for preventing and / or treating tumors, cancers, viral infections, organ transplant rejection, neurodegenerative diseases, attention-related diseases or autoimmune diseases, which comprises administering the compound of the present invention or the compound or pharmaceutical composition of the present invention to a mammal in need thereof.

[0056] The pharmaceutical composition of the present invention can be formulated into dosage forms for oral administration or parenteral administration (including intramuscular, intravenous and subcutaneous routes, intratumor injection) according to any of the conventional methods, such as tablets, granules, powders, capsules, syrups, emulsions, microemulsions, solutions or suspensions.

[0057] The pharmaceutical composition of the present invention for oral administration can be prepared by mixing the active ingredient with a carrier such as cellulose, calcium silicate, corn starch, lactose, sucrose, dextrose, calcium phosphate, stearic acid, magnesium stearate, calcium stearate, gelatin, talc, surfactants, suspending agents, emulsifiers and diluents.

[0058] Examples of carriers used in the pharmaceutical compositions for injection administration of the present invention may be water, saline solutions, glucose solutions, glucose-like solutions, alcohols, glycols, ethers (e.g., polyethylene glycol 400), oils, fatty acids, fatty acid esters, glycerides, surfactants, suspending agents and emulsifiers.

[0059] Other features of the invention will become apparent as the present invention proceeds through the description of exemplary embodiments, which are given to illustrate the invention and are not intended to be limiting thereof. The following examples were prepared, isolated, and characterized using the methods disclosed herein.

[0060] The compounds of the present invention can be prepared in a variety of ways known to those skilled in the art of organic synthesis, and the compounds of the present invention can be synthesized using the following methods and synthetic methods known in the field of organic synthetic chemistry or by variations thereof known to those skilled in the art. Preferred methods include, but are not limited to, those described below. The reaction is carried out in a solvent or solvent mixture suitable for the kit materials used and suitable for the transformation achieved. It will be understood by those skilled in the art of organic synthesis that the functionality present on the molecule is consistent with the proposed transformation. This sometimes requires judgment to change the order of the synthesis steps or the raw materials to obtain the desired compounds of the present invention. DETAILED DESCRIPTION

[0061] the term

[0062] If not otherwise stated, the terms used in this application, including the specification and claims, are defined as follows. If not otherwise stated, conventional methods of mass spectrometry, nuclear magnetic resonance, HPLC, protein chemistry, biochemistry, recombinant DNA technology and pharmacology are used. In this application, if not otherwise stated, the use of "or" or "and" means "and / or".

[0063] In the specification and claims, a given chemical formula or name shall encompass all stereoisomers and optical isomers thereof and racemates in which such isomers exist. Unless otherwise indicated, all chiral (enantiomers and diastereomers) and racemic forms are within the scope of the present invention. Multiple geometric isomers of C=C double bonds, C=N double bonds, ring systems, etc. may also exist in the compounds, and all such stable isomers are encompassed by the present invention. The present invention describes cis- and trans- (or E- and Z-) geometric isomers of the compounds of the present invention, and they can be separated into mixtures of isomers or separated isomeric forms. The compounds of the present invention can be separated in optically active or racemic form. All methods for preparing the compounds of the present invention and the intermediates prepared therein are considered to be part of the present invention. When enantiomeric or diastereomeric products are prepared, they can be separated by conventional methods (e.g., by chromatography or fractional crystallization). Depending on the process conditions, the final product of the present invention is obtained in free (neutral) or salt form. Both the free forms and salts of these end products are within the scope of the present invention. If desired, one form of the compound may be converted into another form. A free base or acid may be converted into a salt; a salt may be converted into a free compound or another salt; a mixture of isomeric compounds of the present invention may be separated into individual isomers. The compounds of the present invention, their free forms and salts may exist in a variety of tautomeric forms, in which hydrogen atoms are transposed to other parts of the molecule and the chemical bonds between the atoms of the molecule are rearranged. It should be understood that all tautomeric forms that may exist are included in the present invention.

[0064] Unless otherwise defined, the definitions of substituents of the present invention are independent of each other and not related to each other. For example (listing but not exhaustive), in one aspect, for R in a substituent a (or R a '), they are independent of each other in the definitions of different substituents. Specifically, for R a (or R a ') When a definition is selected for a substituent, it does not mean that the R a (or R a ') has the same definition in other substituents. More specifically, for example (only non-exhaustive) for NR a R a ', when R a (or R a ') is selected from hydrogen, it does not mean that in -C(O)-NR a R a ', R a (or R a ') must be hydrogen. In another aspect, when more than one R is present in a substituent a (or Ra '), these R a (or R a ') are also independent of each other. For example, in the substituent -(CR a R a ') m -O-(CR a R a ') n -, when m+n is greater than or equal to 2, the m+n R a (or R a ') are independent of each other and can have the same or different meanings.

[0065] Unless otherwise defined, when a substituent is noted as "optionally substituted", the substituent is selected, for example, from substituents such as alkyl, cycloalkyl, aryl, heterocyclyl, halogen, hydroxy, alkoxy, oxo, alkanoyl, aryloxy, alkanoyloxy, amino, alkylamino, arylamino, arylalkylamino, disubstituted amino (wherein the two amino substituents are selected from alkyl, aryl or arylalkyl), alkanoylamino, aroylamino, aralkanoylamino, substituted alkanoylamino, substituted arylamino, substituted aralkanoylamino, thio, alkylthio, arylthio, arylalkylthio, arylthiocarbonyl, arylalkylthiocarbonyl, alkylsulfonyl, arylsulfonyl, arylalkylsulfonyl, aminosulfonyl, for example -SO 2 NH 2 , substituted sulfonylamino, nitro, cyano, carboxyl, carbamoyl such as -CONH 2 , substituted carbamoyl such as -CONHalkyl, -CONHR, -CONHRalkyl or having two substituents on nitrogen selected from alkyl, aryl or arylalkyl, alkoxycarbonyl, aryl, substituted aryl, guanidinyl, heterocyclic groups such as indolyl, imidazolyl, furanyl, thienyl, thiazolyl, pyrrolidinyl, pyridinyl, pyrimidinyl, pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, homopiperazinyl and the like and substituted heterocyclic groups.

[0066] As used herein, the term "alkyl" or "alkylene" is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms. 1 -C 6 The term "alkyl" refers to an alkyl group having 1 to 6 carbon atoms. Examples of the alkyl group include, but are not limited to, methyl (Me), ethyl (Et), propyl (e.g., n-propyl and isopropyl), butyl (e.g., n-butyl, isobutyl, tert-butyl), and pentyl (e.g., n-pentyl, isopentyl, neopentyl). In this context, the alkyl group is preferably an alkyl group having 1 to 6, 1 to 4, more preferably 1 to 3 carbon atoms.

[0067] The term "alkenyl" refers to a straight or branched chain hydrocarbon group containing one or more double bonds and generally having a length of 2 to 20 carbon atoms. 2 -C 6 "Alkenyl" contains two to six carbon atoms. Alkenyl includes, but is not limited to, for example, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, and the like.

[0068] The term "alkynyl" refers to a straight or branched chain hydrocarbon group containing one or more triple bonds and generally having a length of 2 to 20 carbon atoms. 2 -C 6 "Alkynyl" contains two to six carbon atoms. Representative alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 1-butynyl, and the like.

[0069] The term "alkoxy" or "alkyloxy" refers to an -O-alkyl group. 1 -C 6 Alkoxy" (or alkyloxy) is intended to include C 1 , C 2 , C 3 , C 4 , C 5 , C 6 Alkoxy. Examples of alkoxy include, but are not limited to, methoxy, ethoxy, propoxy (e.g., n-propoxy and isopropoxy), and tert-butoxy. In this context, alkoxy is preferably an alkoxy having 1 to 6, more preferably 1 to 4 carbon atoms. Similarly, "alkylthio" or "thiothio" means an alkyl group as defined above connected via a sulfur bridge having a specified number of carbon atoms; for example, methyl-S- and ethyl-S-.

[0070] The term "carbonyl" refers to an organic functional group consisting of carbon and oxygen atoms connected by a double bond (C=O).

[0071] The term "aryl", alone or as part of a larger moiety such as "aralkyl", "arylalkoxy" or "aryloxyalkyl", refers to a monocyclic, bicyclic or tricyclic ring system having a total of 5 to 12 ring members, wherein at least one ring in the system is aromatic and wherein each ring in the system contains 3 to 7 ring members. In certain embodiments of the present invention, "aryl" refers to an aromatic ring system, which includes but is not limited to phenyl, biphenyl, indanyl, 1-naphthyl, 2-naphthyl and tetrahydronaphthyl. The term "aralkyl" or "arylalkyl" refers to an alkyl residue attached to an aryl ring, non-limiting examples of which include benzyl, phenethyl and the like. The fused aryl group may be attached to another group at a suitable position on the cycloalkyl ring or the aromatic ring. The dotted line drawn from the ring system indicates that the bond may be attached to any suitable ring atom.

[0072] The term "cycloalkyl" refers to a monocyclic or bicyclic cyclic alkyl group. A monocyclic cyclic alkyl group refers to a C3 -C 8 Cyclic alkyl includes but is not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and norbornyl. Branched cycloalkyl such as 1-methylcyclopropyl and 2-methylcyclopropyl are included in the definition of " cycloalkyl ". Bicyclic cyclic alkyl includes the cycloalkyl of bridge ring, spiro ring or condensed ring. In this article, cycloalkyl is preferably a cycloalkyl with 3 to 8, more preferably with 3 to 6 carbon atoms.

[0073] The term "cycloalkenyl" refers to a monocyclic or bicyclic cyclic alkenyl. A monocyclic cyclic alkenyl refers to a C 3 -C 8 Cyclic alkenyl groups include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl and norbornyl. Branched cycloalkenyl groups such as 1-methylcyclopropenyl and 2-methylcyclopropenyl are included in the definition of "cycloalkenyl". Bicyclic cycloalkenyl groups include cyclic alkenyl groups of bridged rings, spiro rings or condensed rings.

[0074] "Halo" or "halogen" includes fluorine, chlorine, bromine and iodine. "Haloalkyl" is intended to include branched and straight-chain saturated aliphatic hydrocarbon groups with a specified number of carbon atoms and substituted with one or more halogens. Examples of haloalkyl include, but are not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, trichloromethyl, pentafluoroethyl, pentachloroethyl, 2,2,2-trifluoroethyl, heptafluoropropyl and heptachloropropyl. Examples of haloalkyl also include "fluoroalkyl" which is intended to include branched and straight-chain saturated aliphatic hydrocarbon groups with a specified number of carbon atoms and substituted with one or more fluorine atoms. Similarly, "halocycloalkyl" is intended to include branched cycloalkyl groups with a specified number of carbon atoms and substituted with one or more halogens.

[0075] "Haloalkoxy" or "haloalkyloxy" means a haloalkyl group as defined above having the specified number of carbon atoms attached via an oxygen bridge. 1 -C 6 "Alkoxy" is intended to include C 1 , C 2 , C 3 , C 4 , C 5 , C 6 Halogenated alkoxy. Examples of halogenated alkoxy include, but are not limited to, trifluoromethoxy, 2,2,2-trifluoroethoxy, and pentafluoroethoxy. Similarly, "haloalkylthio" or "thiohaloalkoxy" means a haloalkyl group as defined above connected via a sulfur bridge with a specified number of carbon atoms; for example, trifluoromethyl-S- and pentafluoroethyl-S-.

[0076] The halogenated (C 1 -C 6 )alkyleneCN, -halogenated(C 3 -C 8)cycloalkyleneCN, -halogenated(C 1 -C 6 )Alkylene NR a R a ' etc. means that -(C 1 -C 6 )alkyleneCN, -(C 3 -C 8 )cycloalkyleneCN, -(C 1 -C 6 )Alkylene NR a R a '.

[0077] In this disclosure, C is used when referring to certain substituent groups. x1 -C x2 This means that the number of carbon atoms in the substituent group can be x 1 to x 2 For example, C 0 -C 8 means that the group contains 0, 1, 2, 3, 4, 5, 6, 7 or 8 carbon atoms, C 1 -C 8 means that the group contains 1, 2, 3, 4, 5, 6, 7 or 8 carbon atoms, C 2 -C 8 means that the group contains 2, 3, 4, 5, 6, 7 or 8 carbon atoms, C 3 -C 8 means that the group contains 3, 4, 5, 6, 7 or 8 carbon atoms, C 4 -C 8 means that the group contains 4, 5, 6, 7 or 8 carbon atoms, C 0 -C 6 means that the group contains 0, 1, 2, 3, 4, 5 or 6 carbon atoms, C 1 -C 6 means that the group contains 1, 2, 3, 4, 5 or 6 carbon atoms, C 2 -C 6 means that the group contains 2, 3, 4, 5 or 6 carbon atoms, C 3 -C 6 It means that the group contains 3, 4, 5 or 6 carbon atoms.

[0078] In this disclosure, "x" is used when referring to cyclic groups (e.g., aryl, heteroaryl, cycloalkyl, and heterocycloalkyl). 1 -x 2 The term "ring" means that the number of ring atoms in the group can be x 1 to x 2For example, the 3-12-membered cyclic group can be a 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11-, or 12-membered ring, and the number of its ring atoms can be 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; a 3-6-membered ring means that the cyclic group can be a 3-, 4-, 5-, or 6-membered ring, and the number of its ring atoms can be 3, 4, 5, or 6; a 3-8-membered ring means that the cyclic group can be a 3-, 4-, 5-, 6-, 7-, or 8-membered ring, and the number of its ring atoms can be 3, 4, 5, 6, 7, or 8; a 3-9-membered ring means that the cyclic group can be a 3-, 4-, 5-, 6-, 7-, 8-, or 9-membered ring, and the number of its ring atoms can be 3, 4, 5, 6, 7, 8 or 9; 4-7 membered ring means that the cyclic group can be a 4, 5, 6 or 7 membered ring, and the number of its ring atoms can be 4, 5, 6 or 7; 5-8 membered ring means that the cyclic group can be a 5, 6, 7 or 8 membered ring, and the number of its ring atoms can be 5, 6, 7 or 8; 5-12 membered ring means that the cyclic group can be a 5, 6, 7, 8, 9, 10, 11 or 12 membered ring, and the number of its ring atoms can be 5, 6, 7, 8, 9, 10, 11 or 12; 6-12 membered ring means that the cyclic group can be a 6, 7, 8, 9, 10, 11 or 12 membered ring, and the number of its ring atoms can be 6, 7, 8, 9, 10, 11 or 12. The ring atoms can be carbon atoms or heteroatoms, such as heteroatoms selected from N, O and S. When the ring is a heterocycle, the heterocycle may contain 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more ring heteroatoms, such as heteroatoms selected from N, O and S.

[0079] In the present disclosure, one or more halogens may each independently be selected from fluorine, chlorine, bromine and iodine.

[0080] The term "heteroaryl" means a stable, designated number of monocyclic or polycyclic aromatic groups containing heteroatoms, preferably 3, 4, 5, 6, or 7-membered aromatic monocyclic or aromatic bicyclic or 7, 8, 9, 10, 11, 12-membered aromatic polycyclic heterocyclic rings, which are completely unsaturated or partially unsaturated, and which contain carbon atoms and 1, 2, 3 or 4 heteroatoms independently selected from N, O and S; and include any of the following polycyclic groups, wherein any heterocyclic ring defined above is fused to a benzene ring. The heteroaryl herein is preferably a 5 to 12-membered heteroaryl. Nitrogen and sulfur heteroatoms may be optionally oxidized. The nitrogen atom is substituted or unsubstituted (i.e., N or NR, wherein R is H or, if defined, another substituent). The heterocycle may be connected to its side group at any heteroatom or carbon atom that results in a stable structure. If the resulting compound is stable, the heterocyclic group described herein may be substituted on a carbon or nitrogen atom. The nitrogen in the heterocycle may be optionally quaternized. Preferably, when the total number of S and O atoms in the heterocycle exceeds 1, then these heteroatoms are not adjacent to each other. Preferably, the total number of S and O atoms in the heterocycle is not greater than 1. When the term "heterocycle" is used, it is intended to include heteroaryl.Examples of aromatic hetero groups include, but are not limited to, acridinyl, azetidinyl, azinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzoxazolinyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuranyl, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, imidazopyridinyl, indolenyl, dihydroindole , indolizinyl, indolyl, 3H-indolyl, isatinoyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isothiazolopyridinyl, isoxazolyl, isoxazolopyridinyl, methylenedioxyphenyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolopyridinyl, oxazolidinyl, perylene, oxindolyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiol, phenoxazinyl , phthalazinyl, piperazinyl, piperidinyl, piperidonyl, 4-piperidonyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolopyridinyl, pyrazolyl, pyridazinyl, pyridooxazolyl, pyridoimidazolyl, pyridothiazolyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2-pyrrolidinonyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolyl, 4H-quinolizinyl, quinoxalinyl, quinuclidine, tetrazolyl, tetrahydrofuranyl, tetrahydroisoquinolyl, tetrahydroquinolyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl 1,2,3,4-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl and xanthenyl, quinolyl, isoquinolyl, phthalazinyl, quinazolinyl, indolyl, isoindolyl, indolinyl, 1H-indazolyl, benzimidazolyl, 1,2,3,4-tetrahydroquinolyl, 1,2,3,4-tetrahydroisoquinolyl, 5,6,7,8-tetrahydro-quinolyl, 2,3-dihydro-benzofuranyl, chromanyl, 1,2,3,4-tetrahydro-quinoxalinyl and 1,2,3,4-tetrahydro-quinazolinyl.The term "heteroaryl" may also include biaryl structures formed by the "aryl" defined above and a monocyclic "heteroaryl", such as but not limited to "-phenylbipyridyl-", "-phenylbipyrimidyl-", "-pyridylbiphenyl-", "-pyridylbipyrimidyl-", "-pyrimidylbiphenyl-"; the present invention also includes fused ring and spiro compounds containing, for example, the above-mentioned heterocycles.

[0081] The term "heterocycloalkyl" as used herein refers to a monocyclic heterocycloalkyl system, or a bicyclic heterocycloalkyl system, and also includes spiro heterocyclic or bridged heterocycloalkyl. In the present invention, a monocyclic heterocycloalkyl refers to a 3-8 membered saturated or unsaturated but non-aromatic cyclic alkyl system containing at least one heteroatom selected from O, N, S and P. A bicyclic heterocycloalkyl system refers to a heterocycloalkyl fused to a phenyl, or a cycloalkyl, or a cycloalkenyl, or a heterocycloalkyl, or a heteroaryl to form a bicyclic system.

[0082] The term "bridged cycloalkyl" used herein refers to a polycyclic compound that shares two or more carbon atoms. It can be divided into bicyclic bridged hydrocarbons and polycyclic bridged hydrocarbons. The former is composed of two alicyclic rings sharing two or more carbon atoms; the latter is a bridged hydrocarbon composed of three or more rings.

[0083] The term "spirocycloalkyl" as used herein refers to a polycyclic hydrocarbon group in which the single rings share a carbon atom (called a spiro atom).

[0084] The term "bridged heterocyclic group" as used herein refers to a polycyclic compound that shares two or more atoms, and the ring contains at least one heteroatom selected from O, N and S. It can be divided into bicyclic bridged heterocyclic rings and polycyclic bridged heterocyclic rings.

[0085] The term "heterospirocyclic group" as used herein refers to a polycyclic hydrocarbon group in which single rings share a carbon atom (called a spiro atom), and the ring contains at least one heteroatom selected from O, N and S atoms.

[0086] The term "substituted" as used herein means that at least one hydrogen atom is replaced by a non-hydrogen group, provided that normal valence is maintained and the substitution results in a stable compound. As used herein, a ring double bond is a double bond formed between two adjacent ring atoms (e.g., C=C, C=N, or N=N).

[0087] In the case of nitrogen atoms (e.g., amines) present on the compounds of the invention, these nitrogen atoms can be converted to N-oxides by treatment with an oxidizing agent (e.g., mCPBA and / or hydrogen peroxide) to obtain other compounds of the invention. Thus, the nitrogen atoms shown and claimed are considered to encompass both the shown nitrogen and its N-oxide to obtain the derivatives of the invention.

[0088] When any variable occurs more than one time in any constituent or formula for a compound, its definition at each occurrence is independent of its definition at every other occurrence. Thus, for example, if a group is shown substituted with 0-3 R, then the group may be optionally substituted with up to three R groups, and at each occurrence R is independently selected from the definition of R. Furthermore, combinations of substituents and / or variables are permissible only if such combinations result in stable compounds.

[0089] The term "patient" as used herein refers to an organism treated by the method of the present invention. Such organisms preferably include, but are not limited to, mammals (e.g., mice, apes, monkeys, horses, cows, pigs, dogs, cats, etc.) and most preferably refer to humans.

[0090] The term "effective amount" as used herein means the amount of a drug or medicament (i.e., a compound of the present invention) that will cause a biological or medical response of a tissue, system, animal, or human being, such as that sought by a researcher or clinician. In addition, the term "therapeutically effective amount" means an amount that results in improved treatment, cure, prevention, or alleviation of a disease, condition, or side effect, or reduces the rate of progression of a disease or condition, compared to a corresponding subject that has not received the above amount. An effective amount can be given in one or more administrations, applications, or doses and is not intended to be limited by a specific formulation or route of administration. The term also includes an effective amount that enhances normal physiological function within its scope.

[0091] As used herein, the term "treat," ...

[0092] The terms "pharmaceutically" or "pharmaceutically acceptable" are used herein to refer to those compounds, substances, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response and / or other problems or complications, and commensurate with a reasonable benefit / risk ratio.

[0093] As used herein, the phrase "pharmaceutically acceptable carrier" means a pharmaceutical substance, composition or vehicle, such as a liquid or solid filler, diluent, excipient, manufacturing aid (e.g., lubricant, talc, magnesium, calcium or zinc stearate or stearic acid) or solvent encapsulating substance, which is involved in carrying or transporting the subject compound from one organ or part of the body to another. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient.

[0094] The term "pharmaceutical composition" means a composition comprising a compound of the present invention and at least one other pharmaceutically acceptable carrier. "Pharmaceutically acceptable carrier" refers to a medium generally accepted in the art for delivering biologically active agents to animals (particularly mammals), including (i.e.) adjuvants, excipients or vehicles, such as diluents, preservatives, fillers, flow regulators, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, aromatics, antibacterial agents, antifungal agents, lubricants and dispersants, depending on the nature of the mode of administration and dosage form.

[0095] Specific pharmaceutical and medical terms

[0096] The term "acceptable," as used herein, means that a formulation component or active ingredient has no undue deleterious effect on health and well-being for the general purpose of treatment.

[0097] The term "cancer", as used herein, refers to an abnormal growth of cells that cannot be controlled and can metastasize (spread) under certain conditions. This type of cancer includes, but is not limited to, solid tumors (such as bladder, intestine, brain, chest, uterus, heart, kidney, lung, lymphoid tissue (lymphoma), ovary, pancreas or other endocrine organs (such as thyroid), prostate, skin (melanoma) or blood tumors (such as non-leukemic leukemia).

[0098] The term "combination administration" or its like, as used herein, refers to the administration of several selected therapeutic agents to one patient, in the same or different administration routes and at the same or different times.

[0099] The term "enhance" or "enhancing", as used herein, refers to the ability to increase or prolong the effectiveness or duration of a desired result. Therefore, in terms of enhancing the therapeutic effect of a drug, the term "enhancing" refers to the ability of a drug to increase or prolong the effectiveness or duration in a system. "Efficacy value" as used herein refers to the ability to maximize the enhancement of another therapeutic drug in an ideal system.

[0100] The term "immune disease" refers to a disease or condition that is caused by an adverse or harmful response to endogenous or exogenous antigens. The result is usually cellular dysfunction, or the resulting damage and malfunction of, or destruction of, organs or tissues that may produce immune symptoms.

[0101] The terms "kit" and "product package" are synonymous.

[0102] The term "subject" or "patient" includes mammals and non-mammals. Mammals include, but are not limited to, mammals: humans, non-human primates such as gorillas, apes and monkeys; agricultural animals such as cattle, horses, goats, sheep, pigs; livestock such as rabbits and dogs; experimental animals include rodents such as rats, mice and guinea pigs. Non-mammalian animals include, but are not limited to, birds, fish, etc. In a preferred aspect, the selected mammal is a human.

[0103] The terms "treat," "treatment," or "therapy" as used herein include alleviating, inhibiting, or ameliorating symptoms or conditions of a disease; inhibiting the development of complications; ameliorating or preventing underlying metabolic syndrome; inhibiting the development of a disease or symptom, such as controlling the progression of a disease or condition; alleviating a disease or symptom; causing a disease or symptom to regress; alleviating complications caused by a disease or symptom, or preventing and / or treating signs caused by a disease or symptom.

[0104] As used herein, a compound or pharmaceutical composition, after administration, can improve a disease, symptom or condition, especially improve its severity, delay the onset, slow the progression of the disease, or reduce the duration of the disease. Whether fixed or temporary administration, continuous or intermittent administration, can be attributed to or related to the administration.

[0105] Route of administration

[0106] Suitable routes of administration include, but are not limited to, oral, intravenous, rectal, aerosol, parenteral, ocular, pulmonary, transdermal, vaginal, auricular, nasal, and topical administration. In addition, parenteral administration, by way of example only, includes intramuscular, subcutaneous, intravenous, intramedullary, ventricular, intraperitoneal, intralymphatic, and intranasal injections.

[0107] On the one hand, the administration of the compounds described herein is local rather than systemic. In a specific embodiment, the long-acting formulation is administered by implantation (e.g., subcutaneous or intramuscular) or by intramuscular injection. In addition, in another embodiment, the drug is administered by a targeted drug delivery system. For example, a liposome wrapped by an organ-specific antibody. In this embodiment, the liposome is selectively directed to a specific organ and absorbed.

[0108] Example

[0109] General process

[0110] When the preparation route is not included, the raw materials and reagents used in the present invention are all known products, which can be synthesized according to methods known in the art, or can be obtained by purchasing commercial products. The commercial reagents used do not need to be further purified. Room temperature refers to 20-30°C.

[0111] Unless otherwise specified in the reaction examples, the reactions were carried out under a nitrogen atmosphere, which means that the reaction flask was connected to a nitrogen balloon of about 1 L.

[0112] The hydrogenation reaction is usually carried out by evacuating the vacuum and filling with hydrogen, and the operation is repeated three times. The hydrogen atmosphere means that the reaction bottle is connected to a hydrogen balloon of about 1L.

[0113] Microwave reaction use Initiator + microwave reactor.

[0114] The structures of the compounds of the present invention were determined by nuclear magnetic resonance (NMR) and mass spectrometry (MS). -6 The units of (ppm) are given. NMR measurements were performed using (Bruker Ascend TM 500) NMR, the solvents used for the determination were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (CD 3 OD), with tetramethylsilane (TMS) as the internal standard. The following abbreviations are used for the multiplicity of NMR signals: s = singlet, brs = broad, d = doublet, t = triplet, m = multiplet. Coupling constants are listed as J values, measured in Hz.

[0115] The LC-MS was determined using a Thermo liquid mass spectrometer (UltiMate 3000+MSQ PLUS). The HPLC was determined using a Thermo high pressure liquid chromatograph (UltiMate 3000). The reversed phase preparative chromatography was performed using a Thermo (UltiMate 3000) reversed phase preparative chromatograph. The rapid column chromatography was performed using an Agilent (FS-9200T) automatic column machine, and the silica gel prepacked column was performed using a Sanqin Pre-packed column. The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate, and the specification used for thin layer chromatography separation and purification products is 0.4mm~0.5mm.

[0116] The synthetic methods of some intermediates in the present invention are as follows:

[0117] Intermediate 1

[0118]

[0119] Intermediate 1 was prepared by the following steps:

[0120]

[0121] Step 1: Dissolve compound Int-1a (5.0 g, 25.00 mmol), Int-1b (3.87 g, 32.50 mmol) and N, N-diisopropylethylamine (9.69 g, 74.99 mmol) in tetrahydrofuran (50 mL) and react at room temperature overnight. After the reaction is completed by LCMS monitoring, water (100 mL) is added, filtered, the filter cake is washed with water, and then dried to obtain a white solid Int-1c (5.5 g, yield 77%). ESI-MS (m / z): 283.2 [M+H] + .

[0122] Step 2: Compound Int-1c (5.5 g, 19.46 mmol), platinum dioxide (441 mg, 1.95 mmol) and hydrochloric acid-1,4-dioxane solution (4.86 mL, 19.46 mmol, 4 M) were dissolved in tetrahydrofuran (50 mL), replaced with a hydrogen balloon three times and reacted at room temperature for 48 hours. After the reaction was completed by LCMS monitoring, methanol (100 mL) was added for dilution, filtered, the filter cake was washed with methanol, the filtrate was alkalized with ammonia methanol (7 M) (pH = 9-10), then filtered, the filter cake was washed with water, and then dried to obtain a gray solid Int-1d (3.0 g, yield 60%). ESI-MS (m / z): 255.2 [M+H] + .

[0123] Step 3: Add compound Int-1d (1.0 g, 3.93 mmol) and cesium carbonate (2.56 g, 7.85 mmol) to a 100 mL single-necked flask, add acetonitrile (20 mL), then dropwise add iodomethane (585 mg, 4.12 mmol), and react at room temperature overnight. After the reaction is completed, the reaction solution is diluted with ethyl acetate (50 mL) and filtered, and the filter cake is washed with ethyl acetate. The filtrate is concentrated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1) to obtain a yellow solid Int-1 (800 mg, yield 75%). ESI-MS (m / z): 269.2 [M+H] + .

[0124] Intermediate 2

[0125]

[0126] Intermediate 2 was prepared by the following steps:

[0127]

[0128] Step 1: Add compound Int-1d (2.0 g, 7.85 mmol) and cesium carbonate (5.12 g, 15.71 mmol) to a 100 mL single-necked flask, add acetonitrile (30 mL), then dropwise add deuterated iodomethane (3.42 g, 23.56 mmol), and react at room temperature overnight. After the reaction is completed by LCMS monitoring, the reaction solution is diluted with ethyl acetate (50 mL), filtered, and the filter cake is washed with ethyl acetate. The filtrate is concentrated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1) to obtain a yellow solid Int-2 (1.3 g, yield 60%). ESI-MS (m / z): 272.2 [M+H] + .

[0129] Intermediate 3

[0130]

[0131] Intermediate 3 was prepared by the following steps:

[0132]

[0133] Step 1: Dissolve Int-3a (5.0 g, 41.97 mmol) in methanol (50 mL), place in an ice-water bath, slowly drop thionyl chloride (14.99 g, 125.92 mmol, 9.14 mL), warm to room temperature after the addition is complete, and then to 60 ° C to react overnight. After the reaction is completed by LCMS monitoring, the reaction solution is concentrated to obtain a white solid Int-3b (6.5 g, yield 91%). ESI-MS (m / z): 170.2 [M+H] + .

[0134] Step 2: Dissolve compound Int-1a (7.0 g, 35.00 mmol), Int-3b (6.5 g, 38.50 mmol) and N, N-diisopropylethylamine (13.57 g, 104.99 mmol) in tetrahydrofuran (70 mL) and react at room temperature overnight. After the reaction is completed by LCMS monitoring, water (150 mL) is added, filtered, the filter cake is washed with water, and then dried to obtain a white solid Int-3c (9.0 g, yield 86%). ESI-MS (m / z): 297.2 [M+H] + .

[0135] Step 3: Compound Int-3c (9.0 g, 30.33 mmol), platinum dioxide (688 mg, 3.03 mmol) and hydrochloric acid-1,4-dioxane solution (7.58 mL, 30.33 mmol, 4 M) were dissolved in tetrahydrofuran (100 mL), replaced with a hydrogen balloon three times and reacted at room temperature for 48 hours. After the reaction was completed by LCMS monitoring, methanol (150 mL) was added for dilution, filtered, the filter cake was washed with methanol, the filtrate was alkalized with ammonia methanol (7 M) (pH = 9-10), then filtered, the filter cake was washed with water, and then dried to obtain a gray solid Int-3d (5.5 g, yield 67%). ESI-MS (m / z): 269.3 [M+H] + .

[0136] Step 3: Add compound Int-3d (2.0 g, 7.44 mmol) and cesium carbonate (4.85 g, 14.89 mmol) to a 100 mL single-necked flask, add acetonitrile (30 mL), then dropwise add deuterated iodomethane (1.62 g, 11.16 mmol), and react at room temperature overnight. After the reaction is completed by LCMS monitoring, the reaction solution is diluted with ethyl acetate (100 mL), filtered, and the filter cake is washed with ethyl acetate. The filtrate is concentrated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1) to obtain a yellow solid Int-3 (1.5 g, yield 70%). ESI-MS (m / z): 286.3 [M+H] + .

[0137] Intermediate 4

[0138]

[0139] Intermediate 4 was prepared by the following steps:

[0140]

[0141] Step 1: Add compound Int-4a (700 mg, 3.54 mmol), Int-4b (433 mg, 3.54 mmol) and cesium carbonate (2.31 g, 7.09 mmol) to N, N-dimethylformamide (10 mL), react at 50 ° C for 16 hours, and monitor the reaction by LCMS. Remove N, N-dimethylformamide by distillation under reduced pressure, add water and ethyl acetate for extraction, combine the organic phases and wash with saturated brine, dry over anhydrous sodium sulfate, filter and concentrate the organic phase to obtain a white solid Int-4c (711 mg, yield 67%).

[0142] Step 2: Dissolve compound Int-4c (355 mg, 1.18 mmol) in methanol (35 mL), add ammonia water (3.5 mL) and Raney nickel (3.5 mL, water suspension) to the reaction system in sequence, replace hydrogen with a hydrogen balloon and react for 3 hours under a hydrogen atmosphere and room temperature. LCMS monitors the completion of the reaction. The reaction solution is diluted with methanol, filtered, and the filtrate is concentrated and purified by silica gel column chromatography (dichloromethane: methanol = 9: 1) to obtain a brown oily liquid Int-4 (199 mg, yield 55%). ESI-MS (m / z): 304.2 [M+H] + .

[0143] Intermediate 5

[0144]

[0145] Intermediate 5 was prepared by the following steps:

[0146]

[0147] Step 1: Ethyl formate (3.17 g, 42.73 mmol) and solid sodium ethoxide (3.49 g, 50.50 mmol) were added to tetrahydrofuran (140 mL) in sequence. Compound Int-5a (7.0 g, 38.85 mmol) was added at 5-10°C, the temperature was raised to 50°C and stirred for 2 hours. The raw material disappeared after monitoring by HPLC, and tetrahydrofuran was evaporated under reduced pressure to obtain a yellow oil Int-5b, which was directly used in the next step.

[0148] Step 2: Add 150 mL of anhydrous ethanol to the oily substance Int-5b obtained in the previous step, stir at room temperature to dissolve, add trifluoroacetamide (4.35 g, 33.01 mmol, purity 85%) dropwise, keep stirring at 30°C for 5 hours, then raise the temperature to 80°C and continue stirring for 2 hours. HPLC monitoring shows that the raw material disappears, cool down, evaporate about 100 mL of anhydrous ethanol, add the remaining residue to 300 mL of ice water, adjust the pH to 3 with concentrated hydrochloric acid, stir for 0.5 hour, filter with suction, and dry the filter cake to obtain a yellow solid compound Int-5c (4.37 g, two-step reaction yield 41%, purity 99%). ESI-MS (m / z): 271.4 [M+H] + .

[0149] Step 3: Add compound Int-5c (4.0 g, 14.80 mmol) to 60 mL of acetonitrile, add phosphorus oxychloride (6.81 g, 44.41 mmol) dropwise, stir for 10 minutes after the addition is complete, heat to 80°C, keep stirring for 2 hours, and monitor the complete conversion of the raw materials by HPLC. Remove acetonitrile under reduced pressure, add the residual liquid to 200 mL of ice water, stir for 0.5 hours, and filter to obtain a yellow solid Int-5d (3.9 g, yield 86%, purity 95%).

[0150] Step 4: Compound Int-5d (2.0 g, 6.93 mmol), trimethylcyclotriboroxane (2.61 g, 20.79 mmol), palladium acetate (155 mg, 0.69 mol), potassium phosphate (2.94 g, 13.86 mmol) were added to 1,4-dioxane (150 mL) in sequence, and water (15 mL) was added. Under nitrogen protection, the mixture was stirred at 90°C for 17 hours. HPLC monitored the complete conversion of the raw materials and the filtrate was concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 9) to obtain a white solid Int-5e (1.22 g, yield 65%, purity 99%). ESI-MS (m / z): 269.1 [M+H] + .

[0151] Step 5: Compound Int-5e (1.0 g, 3.73 mmol) was added to 20 ml of methanol, and palladium carbon (10%, 100 mg) was added. The reaction system was replaced with hydrogen and stirred overnight at room temperature. The reaction solution was filtered through diatomaceous earth, and the filtrate was concentrated to obtain compound Int-5f (630 mg, yield 92%, purity 97%). ESI-MS (m / z): 177.1 [MH] - .

[0152] Step 6: Compound Int-5f (0.5 g, 2.81 mmol), Cs 2 CO 3 (1.83 g, 5.61 mmol), Int-4b (514 mg, 4.21 mmol) were added to N, N-dimethylformamide (10 mL) in sequence, stirred at 20°C overnight, and the raw material disappeared after HPLC monitoring. The reaction solution was added to 50 ml of ice water, stirred for 0.5 hours, and filtered to obtain a yellow solid Int-5g (510 mg, yield 64%, purity 99%). ESI-MS (m / z): 281.3 [M+H] + .

[0153] Step 7: Dissolve compound Int-5g (100mg, 0.36mmol) in methanol (5mL), add ammonia water (0.2mL), add Raney Nickel (0.5mL, water suspension), replace hydrogen in the reaction system and stir at room temperature for 2 hours. Filter the reaction solution with diatomaceous earth and concentrate the filtrate. Obtain compound Int-5 (95mg, yield 93%, purity 90%), ESI-MS (m / z): 284.9 [M+H] + .

[0154] Intermediate 6

[0155]

[0156] Intermediate 6 was prepared by the following steps:

[0157]

[0158] Step 1: Add compound Int-5d (550 mg, 1.91 mmol), cyclopropylboronic acid (818 mg, 9.53 mmol), palladium acetate (42 mg, 0.19 mmol), tricyclohexylphosphine (106 mg, 0.38 mmol) and potassium phosphate (1.21 g, 5.72 mmol) to a mixed solution of 1,4-dioxane (50 mL) and water (5 mL). After the reaction system is replaced with nitrogen, react at 110 ° C. under a nitrogen atmosphere for 16 hours. LCMS monitoring of the reaction is complete. The solvent is removed by distillation under reduced pressure, and water and ethyl acetate are added for extraction. The organic phases are combined and washed with saturated brine, dried over anhydrous sodium sulfate, and the organic phase is concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5: 1) to obtain a white solid Int-6a (530 mg, yield 94%). ESI-MS (m / z): 295.3 [M+H] + .

[0159] Step 2: Dissolve compound Int-6a (530 mg, 1.80 mmol) in methanol (10 mL), add palladium carbon (10%, 21 mg) to the reaction system, replace hydrogen with a hydrogen balloon and react for 16 hours under a hydrogen atmosphere at room temperature. LCMS monitors the completion of the reaction. The reaction solution is diluted with methanol, filtered, and the filtrate is concentrated to obtain a brown oily liquid Int-6b (330 mg, yield 89%). ESI-MS (m / z): 203.3 [MH] - .

[0160] Step 3: Add compound Int-6b (330 mg, 1.62 mmol), Int-4b (236 mg, 1.94 mmol) and cesium carbonate (1.05 g, 3.23 mmol) to acetonitrile (10 mL), react at room temperature for 16 hours, and monitor the reaction completion by LCMS. Remove acetonitrile by distillation under reduced pressure, then add water and ethyl acetate for extraction, combine the organic phases and wash with saturated brine, dry with anhydrous sodium sulfate, concentrate the organic phase, and purify by silica gel column chromatography (petroleum ether: ethyl acetate = 3: 1) to obtain a white solid Int-6c (450 mg, yield 90%). ESI-MS (m / z): 307.1 [M+H] + .

[0161] Step 4: Dissolve compound Int-6c (350 mg, 1.14 mmol) in methanol (20 mL), add ammonia (3.5 mL) and Raney nickel (3.5 mL, water suspension) to the reaction system in sequence, replace hydrogen with a hydrogen balloon and react for 16 hours under a hydrogen atmosphere at room temperature, and monitor the reaction by LCMS. The reaction solution is diluted with methanol, filtered with diatomaceous earth, and the filtrate is concentrated and purified by silica gel column chromatography (dichloromethane: methanol = 9: 1) to obtain a yellow oily liquid Int-6 (300 mg, yield 84%). ESI-MS (m / z): 311.2 [M+H] + .

[0162] Intermediate 7

[0163]

[0164] Intermediate 7 was prepared by the following steps:

[0165]

[0166] Step 1: Compound Int-5d (2.0 g, 6.93 mmol), Int-7a (3.20 g, 20.79 mmol), palladium acetate (155 mg, 0.69 mol), potassium phosphate (2.94 g, 13.86 mmol) were added to 1,4-dioxane (150 mL) in sequence, and water (15 mL) was added. Under nitrogen protection, the mixture was stirred at 90 ° C for 17 hours. HPLC monitored the complete conversion of the raw materials and the filtrate was concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 9) to obtain a white solid Int-7b (1.18 g, yield 60%, purity 61%). ESI-MS (m / z): 281.1 [M+H] + .

[0167] Step 2: Add compound Int-7b (1.0 g, 3.73 mmol) to 20 ml methanol, add palladium carbon (10%, 100 mg), replace hydrogen in the reaction system and stir overnight at room temperature, filter the reaction solution through diatomaceous earth, and concentrate the filtrate to obtain compound Int-7c (677 mg, yield 94%, purity 97%). ESI-MS (m / z): 191.3 [MH] - .

[0168] Step 3: Compound Int-7c (0.5 g, 2.81 mmol), Cs 2 CO 3 (1.83 g, 5.61 mmol), Int-4b (514 mg, 4.21 mmol) were added to N, N-dimethylformamide (10 mL) in sequence, stirred at 20 ° C overnight, and the raw material disappeared after HPLC monitoring. The reaction solution was added to 50 ml of ice water, stirred for 0.5 hours, and filtered to obtain a yellow solid Int-7d (530 mg, yield 64%, purity 99%). ESI-MS (m / z): 294.3 [M+H] + .

[0169] Step 4: Dissolve compound Int-7d (100 mg, 0.36 mmol) in methanol (5 mL), add ammonia water (0.2 mL), add Raney Nickel (0.5 mL, water suspension), replace hydrogen in the reaction system and stir at room temperature for 2 hours. The reaction solution was filtered through diatomaceous earth, and the filtrate was concentrated to obtain compound Int-7 (97 mg, yield 93%, purity 90%), ESI-MS (m / z): 298.5 [M+H] + .

[0170] Intermediate 8

[0171]

[0172] Intermediate 8 was prepared by the following steps:

[0173]

[0174] Step 1: Dissolve compound Int-8a (4.06 g, 24.89 mmol) and sodium carbonate (5.28 g, 49.79 mmol) in water (80 mL), then add iodine (6.32 g, 24.89 mmol), stir at room temperature for 16 hours, and add saturated sodium thiosulfate (40 mL) and ethyl acetate (40 mL) to the reaction solution. The pH value of the reaction solution was adjusted to 6 by adding concentrated hydrochloric acid, and then extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 4 / 1) to obtain a white solid Int-8b (3.1 g, yield 43%). ESI-MS (m / z): 290.2 [M+H] + .

[0175] Step 2: Compound Int-8b (2.33 g, 6.85 mmol) and potassium carbonate (1.76 g, 10.28 mmol) were dissolved in N, N-dimethylformamide (15 mL), followed by the addition of benzyl bromide (1.42 g, 10.28 mmol), and the mixture was reacted at 50°C for 2 hours. The reaction was monitored by LCMS. Water (20 mL) was then added, and the mixture was extracted with ethyl acetate. The organic phase was washed with a saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1) to obtain a white solid Int-8c (2.1 g, yield 69%) ESI-MS (m / z): 380.2 [M+H] + .

[0176] Step 3: Compound Int-8c (450 mg, 1.18 mmol), tetrahydropyrrole (126 mg, 1.78 mmol), tris(dibenzylideneacetone)dipalladium (108 mg, 0.12 mmol), potassium tert-butoxide (199 mg, 1.78 mmol), 2-dicyclohexylphosphino-2′-(N,N-dimethylamine)-biphenyl (139 mg, 0.36 mmol) were dissolved in toluene. After the reaction system was replaced with nitrogen, the reaction was carried out at 80° C. under a nitrogen atmosphere for 16 hours. The reaction was monitored by LCMS. The reaction solution was filtered with diatomaceous earth to remove insoluble matter, washed with ethyl acetate, and the filtrate was concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate=10 / 1) to obtain yellow oil Int-8d (206 mg, yield 54%). ESI-MS (m / z): 323.3 [M+H] + .

[0177] Step 4: Dissolve compound Int-8d (206 mg, 0.64 mmol) in dichloromethane (5 mL), and lower the temperature of the reaction solution to -78 ° C, then add boron tribromide (801 mg, 3.20 mmol), react at -78 ° C for 2 hours, and monitor the reaction by LCMS. Add water (5 mL), wait for the reaction to warm to room temperature, adjust the pH value of the reaction solution to 6 with 4M sodium hydroxide solution, and extract with dichloromethane. The organic phase is washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered and concentrated, and the residue is purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to obtain a yellow oily liquid Int-8e (120 mg, yield 81%). ESI-MS (m / z): 233.1 [M+H] + .

[0178] Step 5: Dissolve compound Int-8e (100 mg, 0.43 mmol), compound In-4b (63 mg, 0.51 mmol) and cesium carbonate (280 mg, 0.86 mmol) in N,N-dimethylformamide (5 mL), stir at room temperature for 16 hours, and monitor the reaction completion by LCMS. Add water (50 mL), filter under reduced pressure to obtain a light yellow solid Int-8f (103 mg, yield 72%) ESI-MS (m / z): 335.6 [M+H] + .

[0179] Step 6: Compound Int-8f (64 mg, 0.19 mmol) was dissolved in methanol (10 mL), and aqueous ammonia (1 mL) and Raney nickel (3 mL, aqueous suspension) were added to the reaction system in sequence. The hydrogen was replaced by a hydrogen balloon and reacted for 16 hours under a hydrogen atmosphere at room temperature. The reaction was monitored by LCMS to complete. The reaction solution was filtered with diatomaceous earth, and the filtrate was concentrated and purified by silica gel column chromatography (dichloromethane: methanol = 10 / 1) to obtain a brown oily liquid Int-8 (53 mg, yield 84%). ESI-MS (m / z): 339.2 [M+H] + .

[0180] Intermediate 9

[0181]

[0182] Intermediate 9 was prepared by the following steps:

[0183]

[0184] Step 1: Add compound Int-3d (2.0 g, 7.44 mmol) and cesium carbonate (4.85 g, 14.89 mmol) to a 100 mL single-necked flask, add acetonitrile (30 mL), then dropwise add iodomethane (2.11 g, 14.89 mmol), and react at room temperature overnight. After the reaction is completed by LCMS monitoring, the reaction solution is diluted with ethyl acetate (100 mL), filtered, and the filter cake is washed with ethyl acetate. The filtrate is concentrated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1) to obtain a yellow solid Int-9 (1.7 g, yield 80%). ESI-MS (m / z): 283.3 [M+H] + .

[0185] Intermediate 10

[0186]

[0187] Intermediate 10 is prepared by the following steps:

[0188]

[0189] Step 1: At room temperature, triethylamine (2.1 mL, 14.7 mmol) was added dropwise to a dichloromethane solution (15 mL) of compounds Int-10a (1.0 g, 5.9 mmol) and Int-10b (1.1 g, 5.9 mmol). After the reaction solution was stirred at room temperature for 24 hours, LCMS monitored the completion of the reaction. The reaction solution was diluted with dichloromethane, washed with saturated sodium bicarbonate aqueous solution and brine in turn, dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4: 1) to obtain a colorless oily liquid Int-10c (642 mg, yield 50%). ESI-MS (m / z): 220.3 [M+H] + .

[0190] Step 2: Under nitrogen atmosphere and -78°C, potassium bis(trimethylsilyl)amide (2.8 mL, 12.2 mmol) and hexamethylphosphoric triamide (9.6 mL, 54.9 mmol) were added dropwise to a tetrahydrofuran (20 mL) solution of compound Int-10c (2.23 g, 10.2 mmol). After the reaction solution was stirred at -78°C for 30 minutes, iodomethane (1.3 mL, 20.4 mmol) was added dropwise and stirred for 4 hours. The reaction was monitored by LCMS, saturated aqueous ammonium chloride was added to quench the reaction, and ethyl acetate was used for extraction. The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and the organic phases were concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4: 1) to obtain a colorless oily liquid Int-10d (1.7 g, yield 72%). ESI-MS (m / z): 234.3 [M+H]+ Step 3: Add compound Int-10d (2.11 g, 9.0 mmol) to aqueous hydrochloric acid solution (35 mL, 6 M), reflux for 5 hours, and monitor the completion of the reaction by LCMS. Concentrate the reaction solution under reduced pressure to obtain a crude product Int-10e.

[0191] Step 4: The crude product of Int-10e was dissolved in a mixed solvent of methanol (16 mL) and tetrahydrofuran (48 mL), and trimethylsilyldiazomethane (22.6 mL, 45.2 mmol, 2 M inhexanes) was added dropwise under nitrogen atmosphere at 0°C. After reacting at room temperature for 16 hours, the reaction solution was concentrated by vacuum distillation to obtain the crude product Int-10f. ESI-MS (m / z): 147.9 [M+H] + .

[0192] Step 5: The crude product of Int-10f, compound Int-1a (1.09 g, 5.45 mmol) and N,N-diisopropylethylamine (4.75 mL, 27.27 mmol) were dissolved in tetrahydrofuran (15 mL) and reacted at room temperature overnight. After the reaction was completed by LCMS monitoring, tetrahydrofuran was removed by distillation under reduced pressure, extracted with ethyl acetate, the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3: 2) to obtain a slightly yellow solid Int-10g (433 mg, yield 26%). ESI-MS (m / z): 311.1 [M+H] + .

[0193] Step 6: Add compound Int-10g (433 mg, 1.39 mmol), platinum dioxide (32 mg, 0.14 mmol) and hydrochloric acid 1,4-dioxane solution (0.70 mL, 2.79 mmol, 4 M) to tetrahydrofuran (10 mL), replace with hydrogen balloon three times and react at room temperature for 48 hours. After the reaction is completed, methanol is added to dilute the reaction solution, filter, wash the filter cake with methanol, alkalize the filtrate with sodium hydroxide aqueous solution (1 M) (pH = 9-10), extract with ethyl acetate, combine the organic phases and wash with saturated brine, dry with anhydrous sodium sulfate, concentrate the organic phase and purify by silica gel column chromatography (dichloromethane: methanol = 20: 1) to obtain a slightly yellow solid Int-10h (338 mg, yield 86%). ESI-MS (m / z): 283.3 [M+H] + .

[0194] Step 7: Compound Int-10h (159 mg, 0.56 mmol) and cesium carbonate (366 mg, 1.12 mmol) were added to acetonitrile (5 mL), and then iodomethane (120 mg, 0.84 mmol) was added dropwise thereto, and the mixture was reacted at room temperature overnight. After the reaction was completed by LCMS monitoring, acetonitrile was removed by distillation under reduced pressure, and the mixture was extracted with ethyl acetate. The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and the organic phase was concentrated to obtain a yellow solid Int-10. ESI-MS (m / z): 297.3 [M+H] + .

[0195] Intermediate 11

[0196]

[0197] Intermediate 11 is prepared by the following steps:

[0198]

[0199] Step 1: Add a solution of benzyl alcohol (222 mg, 2.05 mmol) in dimethyl sulfoxide (1 mL) dropwise to a suspension of compound Int-11a (500 mg, 2.05 mmol) and sodium hydrogen (123 mg, 3.07 mmol, 60% in oil) in dimethyl sulfoxide (10 mL), and react at room temperature overnight. After the reaction is completed, water is added to quench the reaction, and ethyl acetate is extracted. The organic phases are combined and washed with saturated brine, dried over anhydrous sodium sulfate, and the organic phase is concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 9: 1) to obtain a white solid Int-11b (487 mg, yield 72%). ESI-MS (m / z): 332.1 [M+H] + .

[0200] Step 2: Boron tribromide (0.42 mL, 4.40 mmol) was added dropwise to a dichloromethane (15 mL) solution of compound Int-11b (487 mg, 1.47 mmol) at -78°C and reacted overnight under this condition. After the reaction was completed by LCMS monitoring, methanol was added to quench the reaction, and dichloromethane was extracted. The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and the organic phase was concentrated to obtain a white solid Int-11c. ESI-MS (m / z): 242.3 [M+H] + .

[0201] Step 3: Add compound Int-11c (355 mg, 1.47 mmol), Int-4b (197 mg, 1.61 mmol) and cesium carbonate (956 mg, 2.93 mmol) to N, N-dimethylformamide (10 mL) and react at 50°C for 24 hours. The raw material Int-11c still remains. Add water to quench the reaction, extract with ethyl acetate, combine the organic phases and wash with saturated brine, dry with anhydrous sodium sulfate, concentrate the organic phase, and purify it by silica gel column chromatography (petroleum ether: ethyl acetate = 5: 1) to obtain a white solid Int-11d (150 mg, yield 30%). ESI-MS (m / z): 345.1 [M+H] + .

[0202] Step 4: Under nitrogen atmosphere, borane (1.53 mL, 1.53 mmol, 1N in THF) was added dropwise to a tetrahydrofuran (6 mL) solution of compound Int-11d (150 mg, 0.44 mmol), and the reaction was refluxed for 5 hours. After the reaction was completed by LCMS monitoring, methanol was added to quench the reaction. After the reaction solution was concentrated, it was purified by silica gel column chromatography (dichloromethane: methanol = 10: 1) to obtain a yellow transparent oily liquid Int-11 (74 mg, yield 49%). ESI-MS (m / z): 348.2 [M+H] + .

[0203] Intermediate 12

[0204]

[0205] Intermediate 12 is prepared by the following steps:

[0206]

[0207] Step 1: Dissolve Int-8c (450 mg, 1.18 mmol), dimethylamine hydrochloride (145 mg, 1.78 mmol), tris(dibenzylideneacetone)dipalladium (108 mg, 0.12 mmol), potassium tert-butoxide (400 mg, 3.56 mmol), 2-dicyclohexylphosphino-2′-(N,N-dimethylamine)-biphenyl (139 mg, 0.36 mmol) in toluene. After the reaction system is replaced with nitrogen, react at 80°C under a nitrogen atmosphere for 16 hours. The reaction is monitored by LCMS. Filter the insoluble matter with diatomaceous earth, rinse with ethyl acetate, and concentrate the filtrate. The residue is purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1) to obtain a yellow oily liquid Int-12a (224 mg, yield 54%). MS (m / z): 297.2 [M+H] + .

[0208] Step 2: Dissolve compound Int-12a (151 mg, 0.51 mmol) in dichloromethane (10 mL), and lower the temperature of the reaction solution to -78 ° C, then add boron tribromide (640 mg, 2.55 mmol), react at -78 ° C for 2 hours, and monitor the reaction by LCMS. Add water (10 mL), wait for the reaction to warm to room temperature, adjust the pH value of the reaction solution to 6 with 4M sodium hydroxide solution, and extract with dichloromethane. The organic phase is washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered and concentrated, and the residue is purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to obtain a yellow oily liquid Int-12b (98 mg, yield 93%). MS (m / z): 207.4 [M+H] + .

[0209] Step 3: Dissolve Int-12b (98 mg, 0.48 mmol), Int-4b (63 mg, 0.51 mmol) and cesium carbonate (280 mg, 0.86 mmol) in N,N-dimethylformamide (5 mL), stir at room temperature for 16 hours, and monitor the reaction by LCMS. Add water (50 mL), filter under reduced pressure to obtain a light yellow solid Int-12c (102 mg, yield 70%). MS (m / z): 310.2 [M+H] + .

[0210] Step 4: Compound Int-12c (102 mg, 0.36 mmol) was dissolved in methanol (10 mL), and aqueous ammonia (1 mL) and Raney nickel (3 mL, aqueous suspension) were added to the reaction system in sequence. The hydrogen was replaced by a hydrogen balloon and reacted for 16 hours under a hydrogen atmosphere at room temperature. The reaction was monitored by LCMS to complete. The reaction solution was filtered with diatomaceous earth, and the filtrate was concentrated and purified by silica gel column chromatography (dichloromethane: methanol = 10 / 1) to obtain a brown oily liquid Int-12 (106 mg, yield 99%). MS (m / z): 314.3 [M+H] + .

[0211] The synthesis method of the embodiment compounds of the present invention is as follows:

[0212] Example 1

[0213] (S)-2-(((6-((2-chloro-6-(trifluoromethyl)pyridin-3-yl)oxy)pyridin-3-yl)methyl)amino)-5-(hydroxymethyl)-4-(methyl-d3)-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0214]

[0215] Example 1 was prepared by the following steps:

[0216]

[0217] Step 1: Dissolve Int-2 (42 mg, 0.15 mmol), Int-4 (47 mg, 0.15 mmol) and p-toluenesulfonic acid monohydrate (3 mg, 0.02 mmol) in n-butanol (2 mL) and react under microwave at 160°C for 3 hours. The reaction was completed by LCMS monitoring. The reaction solution was directly purified by reverse phase preparative HPLC to obtain a white solid 1 (41 mg, yield 50%). ESI-MS (m / z): 539.3 [M+H] + ; 1 H NMR (500 MHz, DMSO-d 6 )δ8.12-8.03 (m, 3H), 7.92 (dd, J=8.4, 2.4Hz, 1H), 7.23 (d, J=8.4Hz, 1H), 6.96 (s, 1H), 4.99 (t, J=5.6Hz, 1H), 4.45-4.29 (m, 2H), 4.10-3.98(m, 2H), 3.77-3.64(m, 2H), 3.34-3.28(m, 1H), 2.47(t, J=4.3Hz, 2H), 1.93-1.85(m, 1H), 1.81-1.70(m, 1H).

[0218] Example 2

[0219] (S)-2-(((6-((2-chloro-6-(trifluoromethyl)pyridin-3-yl)oxy)pyridin-3-yl)methyl)amino)-5-(hydroxymethyl)-4-methyl-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0220]

[0221] Example 2 was prepared by the following steps:

[0222]

[0223] Step 1: Dissolve Int-1 (41 mg, 0.15 mmol), Int-4 (47 mg, 0.15 mmol) and p-toluenesulfonic acid monohydrate (3 mg, 0.02 mmol) in n-butanol (2 mL) and react under microwave at 160°C for 3 hours. The reaction was completed by LCMS monitoring. The reaction solution was purified by reverse phase preparative HPLC to obtain a white solid 1 (45 mg, yield 55%). ESI-MS (m / z): 536.2 [M+H] + ; 1H NMR (500 MHz, DMSO-d 6 )δ8.13-8.02 (m, 3H), 7.92 (dd, J=8.4, 2.4Hz, 1H), 7.23 (d, J=8.4Hz, 1H), 6.97 (t, 1H), 4.99 (t, J=5.6Hz, 1H), 4.44-4.29 (m, 2H), 4.05 (t, J= 2.6Hz, 1H), 4.04-3.98(m, 1H), 3.77-3.65(m, 2H), 3.34-3.28(m, 1H), 2.95 (s, 3H), 2.49-2.45 (m, 2H), 1.93-1.84 (m, 1H), 1.82-1.70 (m, 1H).

[0224] Example 3

[0225] (S)-2-(((6-((2-chloro-6-(trifluoromethyl)pyridin-3-yl)oxy)pyridin-3-yl)methyl)amino)-5-(hydroxymethyl)-5-methyl-4-(methyl-d3)-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0226]

[0227] Example 3 was prepared by the following steps:

[0228]

[0229] Step 1: Dissolve Int-3 (44 mg, 0.15 mmol), Int-4 (47 mg, 0.15 mmol) and p-toluenesulfonic acid monohydrate (3 mg, 0.02 mmol) in n-butanol (2 mL) and react under microwave at 160°C for 3 hours. The reaction was completed by LCMS monitoring. The reaction solution was purified by reverse phase preparative HPLC to obtain a white solid 1 (34 mg, yield 40%). ESI-MS (m / z): 553.3 [M+H] + ; 1 H NMR (500 MHz, DMSO-d 6 )δ8.14-8.03 (m, 3H), 7.92 (dd, J=8.5, 2.4Hz, 1H), 7.23 (d, J=8.4Hz, 1H), 6.95 (s, 1H), 5.08 (t, J=5.5Hz, 1H), 4.44 -4.31 (m, 2H), 3.76-3.59 (m, 3H), 3.54 (dd, J=11.3, 5.4Hz, 1H), 2.49-2.45 (m, 2H), 1.89-1.76 (m, 2H), 1.32 (s, 3H).

[0230] Example 4

[0231] (S)-5-(Hydroxymethyl)-4-(methyl-d3)-2-(((6-((4-methyl-2-(trifluoromethyl)pyrimidin-5-yl)oxy)pyridin-3-yl)methyl)amino)-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0232]

[0233] Example 4 was prepared by the following steps:

[0234]

[0235] Step 1: Dissolve compound Int-5 (95 mg, 0.33 mmol) in n-butanol (3 mL), add compound Int-2 (89 mg, 0.33 mmol) and p-toluenesulfonic acid monohydrate (3 mg, 0.02 mmol), and stir the reaction solution at 160°C for 3 hours under microwave conditions. After the reaction solution is cooled to room temperature, it is concentrated under reduced pressure, and the residue is purified by reverse phase preparative HPLC to obtain a white solid 4 (31 mg, yield 18%, purity 99%). ESI-MS (m / z): 517.1 [M+H] + ; 1 H NMR (500 MHz, DMSO-d 6 )δ8.87 (s, 1H), 8.08 (s, 1H), 7.97-7.88 (m, 1H), 7.24 (d, J=8.8, 3.2Hz, 1H), 6.95 (s, 1H), 5.03-4.93 (m, 1H ), 4.46-4.28(m, 2H), 4.10-3.96(m, 2H), 3.80-3.63(m, 2H), 2.96(s, 3H), 2.43(s, 3H), 1.95-1.71(m, 2H).

[0236] Example 5

[0237] (S)-5-(Hydroxymethyl)-5-methyl-4-(methyl-d3)-2-(((6-((4-methyl-2-(trifluoromethyl)pyrimidin-5-yl)oxy)pyridin-3-yl)methyl)amino)-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0238]

[0239] Example 5 was prepared by the following steps:

[0240]

[0241] Step 1: Dissolve compound Int-5 (95 mg, 0.33 mmol) in n-butanol (3 mL), add compound Int-3 (94 mg, 0.33 mmol) and p-toluenesulfonic acid monohydrate (3 mg, 0.02 mmol), and stir the reaction solution at 160°C for 3 hours under microwave conditions. After the reaction solution is cooled to room temperature, it is concentrated under reduced pressure, and the residue is purified by reverse phase preparative HPLC to obtain a white solid 5 (28 mg, yield 15%, purity 99%). ESI-MS (m / z): 533.4 [M+H] + ; 1 H NMR (500 MHz, DMSO-d 6 )δ8.87 (s, 1H), 8.08 (d, J = 2.4Hz, 1H), 7.92 (dd, J = 8.5, 2.4Hz, 1H), 7.23 (d, J = 8.4Hz, 1H), 6.93 (s, 1H), 5.06 (t, J = 5.5Hz, 1H), 4.45-4.3 0 (m, 3H), 3.80-3.66 (m, 2H), 3.66-3.58 (m, 1H), 3.59-3.49 (m, 1H), 2.50 (d, J=1.8Hz, 2H), 2.43 (s, 3H), 1.89-1.77 (m, 2H), 1.33 (s, 3H).

[0242] Example 6

[0243] (S)-5-((S)-1-hydroxyethyl)-4,5-dimethyl-2-(((6-((4-methyl-2-(trifluoromethyl)pyrimidin-5-yl)oxy)pyridin-3-yl)methyl)amino)-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0244]

[0245] Example 6 was prepared by the following steps:

[0246]

[0247] Step 1: Dissolve compound Int-5 (95 mg, 0.33 mmol) in n-butanol (3 mL), add compound Int-10 (97 mg, 0.33 mmol) and p-toluenesulfonic acid monohydrate (3 mg, 0.02 mmol), and stir the reaction solution at 160°C for 3 hours under microwave conditions. After the reaction solution is cooled to room temperature, it is concentrated under reduced pressure, and the residue is purified by reverse phase preparative HPLC to obtain a white solid 6 (37 mg, yield 20%, purity 99%). ESI-MS (m / z): 545.2 [M+H] + ;1 H NMR (500 MHz, DMSO-d 6 ) δ8.86 (s, 1H), 8.08 (d, J = 2.3Hz, 1H), 7.92 (dd, J = 8.5, 2.5Hz, 1H), 7.23 (d, J = 8. 4Hz, 1H), 7.00 (s, 1H), 4.85 (d, J=6.4Hz, 1H), 4.46-4.39 (m, 1H), 4.37-4.30 (m, 1 H), 4.06-3.99(m, 1H), 3.81-3.71(m, 1H), 3.31-3.22(m, 2H), 3.05(s, 3H), 2.43( s, 3H), 1.91 (dd, J=6.8, 3.6Hz, 1H), 1.77 (d, J=3.4Hz, 1H), 0.95 (d, J=6.5Hz, 3H).

[0248] Example 7

[0249] (S)-2-(((6-((4-ethyl-2-(trifluoromethyl)pyrimidin-5-yl)oxy)pyridin-3-yl)methyl)amino)-5-(hydroxymethyl)-5-methyl-4-(methyl-d3)-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0250]

[0251] Example 7 was prepared by the following steps:

[0252]

[0253] Step 1: Dissolve compound Int-7 (50 mg, 0.17 mmol) in n-butanol (3 mL), add compound Int-3 (47 mg, 0.17 mmol) and p-toluenesulfonic acid monohydrate (3 mg, 0.02 mmol), and stir the reaction solution at 160°C for 3 hours under microwave conditions. After the reaction solution is cooled to room temperature, it is concentrated under reduced pressure, and the residue is purified by reverse phase preparative HPLC to obtain a white solid 7 (12 mg, yield 13%, purity 99%). ESI-MS (m / z): 548.2 [M+H] + ; 1 H NMR (500 MHz, DMSO-d 6) δ8.87 (s, 1H), 8.08 (d, J = 2.4Hz, 1H), 7.92 (dd, J = 8.4, 2.4Hz, 1H), 7.24 ( d, J=8.4Hz, 1H), 6.93 (s, 1H), 5.06 (t, J=5.5Hz, 1H), 4.41-4.30 (m, 2H), 3. 78-3.66(m, 2H), 3.65-3.60(m, 1H), 3.58-3.50(m, 1H), 2.81-2.71(m, 2H), 2.49-2.43 (m, 2H), 1.90-1.76 (m, 2H), 1.32 (s, 3H), 1.17 (t, J=7.5Hz, 3H).

[0254] Example 8

[0255] (S)-2-(((6-((4-ethyl-2-(trifluoromethyl)pyrimidin-5-yl)oxy)pyridin-3-yl)methyl)amino)-5-(hydroxymethyl)-4-(methyl-d3)-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0256]

[0257] Example 8 was prepared by the following steps:

[0258]

[0259] Step 1: Dissolve compound Int-7 (50 mg, 0.17 mmol) in n-butanol (3 mL), add compound Int-2 (45 mg, 0.17 umol) and p-toluenesulfonic acid monohydrate (3 mg, 0.02 mmol), and stir the reaction solution at 160°C for 3 hours under microwave conditions. After the reaction solution is cooled to room temperature, it is concentrated under reduced pressure, and the residue is purified by reverse phase preparative HPLC to obtain a white solid 8 (20 mg, yield 22%, purity 99%). ESI-MS (m / z): 533.4 [M+H] + ; 1 H NMR (500 MHz, DMSO-d 6)δ8.87 (s, 1H), 8.08 (d, J=2.4Hz, 1H), 7.92 (dd, J=8.4, 2.5Hz, 1H), 7.24 (d, J=8.4Hz, 1H), 6.94 (s, 2H), 4.97 (t, J=5.5Hz, 1H), 4.44-4.28 (m, 2H ), 4.11-3.97 (m, 2H), 3.79-3.65 (m, 2H), 2.76 (q, J=7.5Hz, 2H), 2.49-2. 45 (m, 2H), 1.96-1.83 (m, 1H), 1.82-1.71 (m, 1H), 1.17 (t, J=7.5Hz, 3H).

[0260] Example 9

[0261] (S)-2-(((6-((4-cyclopropyl-2-(trifluoromethyl)pyrimidin-5-yl)oxy)pyridin-3-yl)methyl)amino)-5-(hydroxymethyl)-4-(methyl-d3)-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0262]

[0263] Example 9 was prepared by the following steps:

[0264]

[0265] Step 1: Dissolve Int-2 (50 mg, 0.18 mmol), Int-6 (57 mg, 0.18 mmol) and p-toluenesulfonic acid monohydrate (3 mg, 0.02 mmol) in n-butanol (2 mL) and react under microwave at 160°C for 3 hours. The reaction was completed by LCMS monitoring. The reaction solution was purified by reverse phase preparative HPLC to obtain a white solid 9 (65 mg, yield 60%). ESI-MS (m / z): 546.3 [M+H] + ; 1 H NMR (500MHz, DMSO-d6) δ8.78 (s, 1H), 8.09 (d, J=2.3Hz, 1H), 7.92 (dd, J=8.5, 2 .4Hz, 1H), 7.25 (d, J=8.4Hz, 1H), 6.94 (t, J=6.5Hz, 1H), 4.97 (s, 1H), 4.46-4.3 0 (m, 2H), 4.10-3.99 (m, 2H), 3.71 (q, J=12.0, 11.4Hz, 2H), 2.47 (t, J=6.6Hz, 2H ), 2.29-2.17(m, 1H), 1.94-1.85(m, 1H), 1.81-1.70(m, 1H), 1.19-1.07(m, 4H).

[0266] Example 10

[0267] (S)-2-(((6-((4-cyclopropyl-2-(trifluoromethyl)pyrimidin-5-yl)oxy)pyridin-3-yl)methyl)amino)-5-(hydroxymethyl)-5-methyl-4-(methyl-d3)-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0268]

[0269] Example 10 was prepared by the following steps:

[0270]

[0271] Step 1: Dissolve Int-3 (50 mg, 0.17 mmol), Int-6 (54 mg, 0.17 mmol) and p-toluenesulfonic acid monohydrate (3 mg, 0.02 mmol) in n-butanol (2 mL) and react under microwave at 160°C for 3 hours. The reaction was completed by LCMS monitoring. The reaction solution was purified by reverse phase preparative HPLC to obtain a white solid 10 (43 mg, yield 44%). ESI-MS (m / z): 560.3 [M+H] + ; 1 H NMR (500MHz, DMSO-d6) δ8.79 (d, J=2.1Hz, 1H), 8.09 (d, J=2.6Hz, 1H), 7.92 (dt, J=8.4, 2.9Hz, 1H), 7.26 (dd, J=8.4, 2.3Hz, 1H), 6.95 (s, 1H), 5.15(s, 1H), 4.50-4.28(m, 2H), 3.76-3.52(m, 4H), 2.50-2.44(m, 2H), 2.30-2.18(m, 1H), 1.91-1.77(m, 2H), 1.33(s, 1H), 1.18-1.07(m, 4H).

[0272] Embodiment 11

[0273] (S)-2-(((6-((4-cyclopropyl-2-(trifluoromethyl)pyrimidin-5-yl)oxy)pyridin-3-yl)methyl)amino)-5-((S)-1-hydroxyethyl)-5-methyl-4-(methyl-d3)-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0274]

[0275] Example 11 was prepared by the following steps:

[0276]

[0277] Step 1: Int-10 (50 mg, 0.17 mmol), Int-6 (52 mg, 0.17 mmol) and p-toluenesulfonic acid monohydrate (3 mg, 0.02 mmol) were dissolved in n-butanol (2 mL) and reacted under microwave conditions at 160 ° C for 3 hours. LCMS monitored the completion of the reaction. The reaction solution was purified by reverse phase preparative HPLC to obtain a white solid 11 (11 mg, yield 12%). ESI-MS (m / z): 571.3 [M+H] + ; 1 H NMR (500MHz, DMSO-d6) δ8.78 (s, 1H), 8.09 (d, J=2.3Hz, 1H), 7.93 (dd, J=8.5, 2.4Hz, 1H), 7.27(d, J=8.4Hz, 1H), 7.13(s, 1H), 4.93(s, 1H), 4.48-4.32(m, 2H), 4.10-3.99(m, 1H), 3. 77(q, J=6.4Hz, 1H), 3.29-3.22(m, 1H), 3.06(s, 3H), 2.53(s, 2H), 2.26-2.19(m, 1H), 1.9 8-1.90 (m, 1H), 1.82-1.73 (m, 1H), 1.53 (s, 3H), 1.15-1.08 (m, 4H), 0.96 (d, J=6.5Hz, 3H).

[0278] Example 12

[0279] (S)-2-(((6-((2-bromo-6-(trifluoromethyl)pyridin-3-yl)oxy)pyridin-3-yl)methyl)amino)-5-(hydroxymethyl)-5-methyl-4-(methyl-d3)-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0280]

[0281] Example 12 was prepared by the following steps:

[0282]

[0283] Step 1: Dissolve compound Int-3 (30 mg, 0.10 mmol), Int-11 (40 mg, 0.12 mmol) and p-toluenesulfonic acid monohydrate (2 mg, 0.01 mmol) in n-butanol (2 mL) and react under microwave at 160°C for 3 hours. The reaction was completed by LCMS monitoring. The reaction solution was purified by reverse phase preparative HPLC to obtain a white solid 12 (11 mg, yield 18%). ESI-MS (m / z): 553.3 [M+H] + ; 1 H NMR (500 MHz, DMSO-d 6 )δ8.12-7.98 (m, 3H), 7.92 (dd, J=8.4, 2.4Hz, 1H), 7.24 (d, J=8.4Hz, 1H), 5.13 (t, J=5.3Hz, 1H), 4.39 (t, J=6.5Hz, 2H), 3.77-3.71 (m, 1 H), 3.69 (dd, J=11.3, 5.7Hz, 1H), 3.65-3.59 (m, 1H), 3.55 (dd, J=11.4, 5.3Hz, 1H), 2.50-2.43 (m, 2H), 1.89-1.77 (m, 2H), 1.34 (s, 3H).

[0284] Example 13

[0285] (S)-5-(Hydroxymethyl)-4,5-dimethyl-2-(((6-((2-(pyrrolidin-1-yl)-6-(trifluoromethyl)pyridin-3-yl)oxy)pyridin-3-yl)methyl)amino)-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0286]

[0287] Example 13 was prepared by the following steps:

[0288]

[0289] Step 1: Dissolve compound Int-8 (77 mg, 0.22 mmol), Int-9 (50 mg, 0.17 mmol) and p-toluenesulfonic acid monohydrate (3 mg, 17 umol) in n-butanol (2 mL) and react under microwave at 160°C for 3 hours. The reaction was completed by LCMS monitoring. The reaction solution was purified by reverse phase preparative HPLC to obtain a white solid 13 (44 mg, yield 43%). ESI-MS (m / z): 585.3 [M+H] + ; 1H NMR (500MHz, DMSO-d6) δ8.08 (d, J=2.3Hz, 1H), 7.84 (dd, J=8.4, 2.4Hz, 1H), 7.39 (d, J=7.8Hz, 1H), 7.05 (d, J=8.0Hz, 1H), 7.02 (d, J=8.1Hz, 1H), 6.89 (br s, 1H), 5.06 (t, J=5.5Hz, 1H), 4.43-4.28 (m, 2H), 3.81-3.67 (m, 2H), 3.67-3.59 (m, 1H), 3.57-3 .53 (m, 1H), 3.50-3.40 (m, 4H), 2.94 (s, 3H), 2.55-2.45 (m, 2H), 1.87-1.70 (m, 6H), 1.33 (s, 3H).

[0290] Embodiment 14

[0291] (S)-2-(((6-((4-ethyl-2-(trifluoromethyl)pyrimidin-5-yl)oxy)pyridin-3-yl)methyl)amino)-5-((S)-1-hydroxyethyl)-4,5-dimethyl-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0292]

[0293] Example 14 was prepared by the following steps:

[0294]

[0295] Step 1: Dissolve compound Int-7 (50 mg, 0.17 mmol) in n-butanol (3 mL), add compound Int-10 (50 mg, 0.17 mmol) and p-toluenesulfonic acid monohydrate (3 mg, 0.02 mmol), and stir the reaction solution at 160°C for 3 hours under microwave conditions. After the reaction solution is cooled to room temperature, it is concentrated under reduced pressure, and the residue is purified by reverse phase preparative HPLC to obtain a white solid 14 (22 mg, yield 23%, purity 99%). ESI-MS (m / z): 559.3 [M+H] + ; 1 H NMR (500 MHz, DMSO-d 6)δ8.87 (s, 1H), 8.07 (s, 1H), 7.92 (d, J = 8.1Hz, 1H), 7.25 (d, J = 8.5Hz, 1H), 7.06 (s, 1H ), 4.99-4.84 (m, 1H), 4.47-4.28 (m, 2H), 4.03 (d, J = 13.0Hz, 1H), 3.82-3.70 (m, 1H), 3 .24(t, J=11.8Hz, 1H), 3.04(s, 3H), 2.82-2.69(m, 2H), 2.55-2.48(m, 2H), 2.01-1.86 (m, 1H), 1.84-1.70 (m, 1H), 1.51 (s, 3H), 1.16 (t, J=7.6Hz, 3H), 0.95 (d, J=6.4Hz, 3H).

[0296] Embodiment 15

[0297] (S)-2-(((6-((2-(dimethylamino)-6-(trifluoromethyl)pyridin-3-yl)oxy)pyridin-3-yl)methyl)amino)-5-(hydroxymethyl)-4,5-dimethyl-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0298]

[0299] Example 15 was prepared by the following steps:

[0300]

[0301] Step 1: Dissolve compound Int-12 (99 mg, 0.31 mmol) in n-butanol (2 mL), add compound Int-9 (50 mg, 0.17 mmol) and p-toluenesulfonic acid monohydrate (6 mg, 35 umol), and stir the reaction solution at 160°C for 3 hours under microwave conditions. After the reaction solution is cooled to room temperature, it is concentrated under reduced pressure, and the residue is purified by reverse phase preparative HPLC to obtain a white solid 15 (28 mg, yield 28%). ESI-MS (m / z): 559.3 [M+H] + ; 1H NMR (500MHz, DMSO-d6) δ8.07 (d, J=2.3Hz, 1H), 7.84 (dd, J=8.5, 2.4Hz, 1H), 7.46 (d, J=7.9Hz, 1H), 7.17 (d, J=8.0Hz, 1H), 7.07 (d, J=8.4Hz, 1H), 6.89 (br s, 1H), 5.06 (t, J=5.4Hz, 1H), 4.43-4.31 (m, 2H), 3.78-3.67 (m, 2H), 3.65-3.60 (m, 1H), 3.58-3.52 (m, 1H), 3.35-3.25 (m, 2H), 3.00-2.90 (m, 9H), 2.55-2.45 (m, 2H), 1.90-1.75 (m, 2H), 1.33 (s, 3H).

[0302] Example 16

[0303] (S)-5-(Hydroxymethyl)-4,5-dimethyl-2-(((6-((2-(methylamino)-6-(trifluoromethyl)pyridin-3-yl)oxy)pyridin-3-yl)methyl)amino)-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0304]

[0305] Example 16 was prepared by the following steps:

[0306]

[0307] Step 1: Dissolve Int-8c (500 mg, 1.18 mmol), methylamine hydrochloride (133 mg, 1.98 mmol), tris(dibenzylideneacetone)dipalladium (120 mg, 0.13 mmol), potassium tert-butoxide (591 mg, 5.28 mmol), 2-dicyclohexylphosphino-2′-(N,N-dimethylamine)-biphenyl (155 mg, 0.39 mmol) in toluene. After the reaction system is replaced with nitrogen, react at 80°C under a nitrogen atmosphere for 16 hours. LCMS monitors the completion of the reaction. Filter the insoluble matter with diatomaceous earth, rinse with ethyl acetate, and concentrate the filtrate. The residue is purified by silica gel column chromatography (petroleum ether / ethyl acetate=10 / 1) to obtain a yellow oily liquid 16a (278 mg, yield 73%). MS (m / z): 283.3 [M+H] + .

[0308] Step 2: Dissolve compound 16a (152 mg, 0.54 mmol) in dichloromethane (3 mL), lower the temperature of the reaction solution to -78 ° C, then add boron tribromide (674 mg, 269 mmol), react at -78 ° C for 2 hours, and monitor the reaction by LCMS. Add water (10 mL), wait for the reaction to warm to room temperature, adjust the pH value of the reaction solution to 6 with 4M sodium hydroxide solution, and extract with dichloromethane. The organic phase is washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered and concentrated, and the residue is purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to obtain a yellow oily liquid 16b (81 mg, yield 78%). MS (m / z): 193.4 [M+H] + .

[0309] Step 3: 16b (81 mg, 0.42 mmol), Int-4b (26 mg, 0.21 mmol) and cesium carbonate (137 mg, 0.42 mmol) were dissolved in N,N-dimethylformamide (2 mL), stirred at room temperature for 16 hours, and the reaction was completed after LCMS monitoring. Water (10 mL) was added, and the mixture was filtered under reduced pressure to obtain a light yellow solid 16c (47 mg, yield 38%). MS (m / z): 295.2 [M+H] + .

[0310] Step 4: Compound 16c (47 mg, 0.16 mmol) was dissolved in methanol (10 mL), and aqueous ammonia (1 mL) and Raney nickel (3 mL, aqueous suspension) were added to the reaction system in sequence. The hydrogen was replaced by a hydrogen balloon and reacted for 16 hours under a hydrogen atmosphere at room temperature. The reaction was monitored by LCMS to complete. The reaction solution was filtered through diatomaceous earth, and the filtrate was concentrated and purified by silica gel column chromatography (dichloromethane: methanol = 10 / 1) to obtain a brown oily liquid 16d (45 mg, yield 94%). MS (m / z): 299.2 [M+H] + .

[0311] Step 5: Compound 16d (45 mg, 0.15 mmol) was dissolved in n-butanol (2 mL), and compound Int-9 (42 mg, 0.15 mmol) and p-toluenesulfonic acid monohydrate (3 mg, 0.02 mmol) were added. The reaction solution was stirred at 160°C for 3 hours under microwave conditions. After the reaction solution was cooled to room temperature, it was concentrated under reduced pressure, and the residue was purified by reverse phase preparative HPLC to obtain a white solid 16 (9 mg, yield 13%). ESI-MS (m / z): 545.4 [M+H] + ; 1 H NMR (500 MHz, DMSO-d 6)δ8.08 (d, J=2.4Hz, 1H), 7.85 (dd, J=8.4, 2.5Hz, 1H), 7.31 (d, J=7.8Hz, 1H), 7.08 (d, J=8.4Hz, 1H), 6.97-6.91 (m, 2H), 6.88 (t, J=4.8Hz, 1H), 5.10 (s, 1H), 4.36 (q, J=7.1, 5.5Hz, 2H), 3.80-3.67 (m, 2H), 3.62 (td, J=8.6, 8.0, 3.9Hz, 1H), 3.54 (d, J =11.4Hz, 1H), 2.95 (s, 3H), 2.79 (d, J = 4.6Hz, 3H), 1.92-1.74 (m, 2H), 1.33 (s, 3H).

[0312] Embodiment 17

[0313] (S)-2-(3-((5-(((5-(Hydroxymethyl)-4-methyl-6-carbonyl-5,6,9,10-tetrahydro-4H,8H-pyrido[3,2,1-de]pteridin-2-yl)amino)methyl)pyridin-2-yl)oxy)-6-(trifluoromethyl)pyridin-2-yl)acetonitrile

[0314]

[0315] Example 17 was prepared by the following steps:

[0316]

[0317] Step 1: Add di-tert-butyl carbonic anhydride (1.75 g, 8.02 mmol) dropwise to a dichloromethane (40 mL) solution of Int-4 (2.03 g, 6.68 mmol) and triethylamine (1.39 mL, 10.02 mmol) under ice-water bath conditions. After stirring the reaction solution at room temperature for 16 hours, water was added to quench the reaction. Extract with dichloromethane, combine the organic phases and wash with saturated brine, dry over anhydrous sodium sulfate, concentrate the organic phase, and purify by silica gel column chromatography (petroleum ether: ethyl acetate = 3: 1) to obtain a white solid 17a (1.95 g, yield 72%). ESI-MS (m / z): 403.9 [M+H] + .

[0318] Step 2: Add compound 17a (675 mg, 1.67 mmol), vinylboronic acid pinacol ester (1.29 g, 8.36 mmol), tetrakistriphenylphosphine palladium (194 mg, 0.17 mol), sodium carbonate (354 mg, 3.34 mmol) to a mixed solution of water (3 mL) and 1,4-dioxane (17 mL). After the reaction system was replaced with nitrogen, it was stirred at 120°C under nitrogen protection for 16 hours, and the reaction was completed after LCMS monitoring. The solvent was removed by distillation under reduced pressure, and water and ethyl acetate were added for extraction. The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3: 1) to obtain a white solid 17b (609 mg, yield 92%). ESI-MS (m / z): 396.4 [M+H] + .

[0319] Step 3: Sodium periodate (907 mg, 4.24 mmol) was added to a mixed solution of 17b (559 mg, 1.41 mmol) in tetrahydrofuran (12 mL) and water (3 mL). After the reaction solution was stirred at room temperature for one minute, potassium osmate (41 mg, 0.14 mmol) was added thereto. After the reaction solution was stirred at 40°C for 2 hours, LCMS monitored the end of the reaction. Water and ethyl acetate were added for extraction, the organic phases were combined and washed with sodium thiosulfate aqueous solution and saturated brine in sequence, dried over anhydrous sodium sulfate, and the organic phase was concentrated to obtain a crude product 17c. ESI-MS (m / z): 398.3 [M+H] + .

[0320] Step 4: Sodium borohydride (80 mg, 2.12 mmol) was added to a methanol (15 mL) solution of the crude product 17c under ice-water bath conditions. The reaction solution was stirred for two hours at this temperature. After the reaction was completed, water and ethyl acetate were added for extraction, the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2: 1) to obtain a white solid 17d (359 mg, two-step yield 64%). ESI-MS (m / z): 400.3 [M+H] + .

[0321] Step 5: Dichlorothionyl (0.13 mL, 1.80 mmol) was added dropwise to a solution of 17d (359 mg, 0.90 mmol) in dichloromethane (14 mL). The reaction mixture was stirred at room temperature for 2 hours, and the reaction was completed after LCMS monitoring. The reaction mixture was concentrated by vacuum distillation to obtain the crude product 17e. ESI-MS (m / z): 418.2 [M+H] + .

[0322] Step 6: Trimethylsilyl cyanide (178 mg, 1.80 mmol) was added dropwise to a solution of cesium fluoride (119 mg, 1.80 mmol) in acetonitrile (8 mL) at 10°C. After the reaction solution was stirred for 30 minutes, cesium carbonate (586 mg, 1.80 mmol) and the crude product 17e were added thereto. The reaction solution was heated to 40°C and stirred at this temperature for 16 hours. After the reaction was completed, water and ethyl acetate were added for extraction, the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and the organic phase was concentrated to obtain the crude product 17f. ESI-MS (m / z): 409.3 [M+H] + .

[0323] Step 7: Trifluoroacetic acid (2 mL) was added dropwise to a dichloromethane (8 mL) solution of the crude product 17f. After the reaction solution was stirred at room temperature for 1.5 hours, the reaction was completed by LCMS monitoring. The reaction solution was concentrated by vacuum distillation, and an aqueous sodium hydroxide solution was added to a pH of 8. The mixture was extracted with ethyl acetate, and the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and the organic phase was concentrated to obtain a crude product 17 g. ESI-MS (m / z): 309.3 [M+H] + .

[0324] Step 8: Compound Int-1 (50 mg, 0.19 mmol) was dissolved in n-butanol (2 mL), and compound Int-17g (69 mg, crude product) and p-toluenesulfonic acid monohydrate (7 mg, 37 umol) were added. The reaction solution was stirred at 160°C for 3 hours under microwave conditions. The reaction was completed by LCMS monitoring. The reaction solution was purified by reverse phase preparative HPLC to obtain a white solid 17 (20 mg, four-step reaction yield 4%). ESI-MS (m / z): 541.2 [M+H] + ; 1 H NMR (500 MHz, DMSO-d 6 )δ8.15 (d, J=16.8Hz, 1H), 8.01-7.87 (m, 3H), 7.20 (d, J=8.4Hz, 1H), 6.97 (t , J=6.4Hz, 1H), 4.99 (s, 1H), 4.42 (dd, J=15.1, 6.5Hz, 1H), 4.34 (dd, J=15.2, 6.2Hz, 1H), 4.29(s, 2H), 4.11-3.98(m, 2H), 3.77-3.65(m, 2H), 3.32-3.27(m , 0H), 2.96 (s, 3H), 2.50-2.44 (m, 2H), 1.97-1.84 (m, 1H), 1.83-1.68 (m, 1H).

[0325] Embodiment 18

[0326] (S)-2-(((6-((2-fluoro-6-(trifluoromethyl)pyridin-3-yl)oxy)pyridin-3-yl)methyl)amino)-5-(hydroxymethyl)-5-methyl-4-(methyl-d3)-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0327]

[0328] Example 18 was prepared by the following steps:

[0329]

[0330] Step 1: Dissolve compound 18a (300 mg, 1.82 mmol) in anhydrous tetrahydrofuran (5 mL), cool to -78 ° C under nitrogen protection, slowly add n-butyl lithium (1.19 mL, 1.91 mmol), start to add triisopropyl borate (0.63 mL, 2.73 mmol) after one hour, and monitor the reaction by LCMS after two hours. Slowly add water to quench the reaction, and when the system is heated to 0 ° C, start to add sodium hydroxide aqueous solution (4M, 1.36 mL, 5.45 mmol) and hydrogen peroxide (1 mL), and react at room temperature for 16 hours. The reaction solution is acidified with hydrochloric acid (2 M), extracted three times with dichloromethane, concentrated, and the residue is purified by silica gel column chromatography (dichloromethane: methanol = 10: 1) to obtain a yellow solid 18b (280 mg, yield 85%). MS (m / z): 180.5 [MH] - .

[0331] Step 2: Compound 18b (280 mg, 1.55 mmol), Int-3d (226 mg, 1.86 mmol) and cesium carbonate (1.01 g, 3.09 mmol) were added to acetonitrile (10 mL) and reacted at room temperature for 16 hours. The reaction was monitored by LCMS. The acetonitrile was removed by distillation under reduced pressure, and water and ethyl acetate were added for extraction. The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5: 1) to obtain a yellow oily liquid 18c (270 mg, yield 61%). ESI-MS (m / z): 284.4 [M+H] + .

[0332] Step 3: Compound 18c (270 mg, 0.95 mmol) was dissolved in methanol (30 mL), and aqueous ammonia (3.5 mL) and Raney nickel (3.5 mL, aqueous suspension) were added to the reaction system in sequence. The hydrogen was replaced by a hydrogen balloon and reacted for 3 hours under a hydrogen atmosphere at room temperature. The reaction was monitored by LCMS to complete. The reaction solution was diluted with methanol, filtered, and the filtrate was concentrated to obtain a brown oily liquid 18d (270 mg, yield 98%). ESI-MS (m / z): 271.5 [M+H] + .

[0333] Step 4: Compound 18d (65.33 mg, 0.23 mmol) was dissolved in n-butanol (2 mL), compound Int-3 (50 mg, 0.17 mmol) and p-toluenesulfonic acid monohydrate (3.32 mg, 17 umol) were added, and the reaction solution was stirred at 160°C for 3 hours under microwave conditions. After the reaction solution was cooled to room temperature, it was concentrated under reduced pressure, and the residue was purified by reverse phase preparative HPLC to obtain a white solid 18 (5.27 mg, yield 5%). ESI-MS (m / z): 536.5 [M+H] + ; 1 H NMR (500MHz, DMSO-d6) δ8.20 (t, J=8.6Hz, 1H), 8.09 (d, J=2.1Hz, 1H), 8.00 (d, J=8.0Hz, 1H), 7.92 (dd, J=8.5, 2.1Hz, 1H), 7.23 (d, J=8.5Hz, 1H), 6.98-6.89 (m , 1H), 5.07 (t, J=5.7Hz, 1H), 4.43-4.33 (m, 2H), 3.79-3.68 (m, 2H), 3.66-3.60 (m, 1H), 3.59-3.52 (m, 1H), 2.50-2.46 (m, 2H), 1.89-1.75 (m, 2H), 1.33 (s, 3H).

[0334] According to the synthetic routes and intermediate synthesis methods described in the above examples, the following examples can be obtained.

[0335]

[0336]

[0337]

[0338] Wnt pathway inhibitor biological screening and results

[0339] Experimental Example 1: Construction of Colo205-LUC-TCF / LEF-M1 reporter cell line

[0340] The Colo205 cell line (Cell Bank of the Chinese Academy of Sciences, Cat#TCHu102) was purchased from the Cell Bank of the Chinese Academy of Sciences. After amplification and subculture, the cells were transfected with a luciferase reporter plasmid (Promega) driven by TCF / LEF transcription factors by lipo3000 liposome transfection during the exponential growth phase of the cells. This plasmid carries a resistance gene and can be screened for resistance. The transfection was carried out in a 10 cm culture dish using a conventional complete medium without resistance. After 2 days, the medium with resistance was replaced and culture continued. After that, the resistance medium was replaced every 2 days, and the suspended cells were discarded. The original medium was centrifuged to remove cells and debris and retained as an adaptive medium. When the cells filled the culture dish, the cells were digested, counted, and subcultured in a 96-well plate so that the average number of cells in each well was 1.5 / well. The adaptation medium was used during subculture. The remaining cells were frozen. After subculture, the cells were cultured for 4 hours to allow the cells to adhere to the wall, and then the number of cells in each well was observed under a microscope. The wells with only one cell in each well are marked, which are monoclonal wells. Then culture normally, change the culture medium every 2 days, and observe. In the early stage, the wells where monoclonal cells continue to grow are marked twice and can be replaced with normal resistant culture medium. When a monoclonal well fills the wells of the 96-well plate, it is digested and subcultured to a 24-well culture plate. After the 24-well plate is full, it is subcultured to a 96-well plate and a 6-well plate. There are at least 6 wells of cells in the 96-well plate, of which 3 wells are added with known Wnt inhibitors, and the other 3 wells are not treated. After 24 hours, the fluorescence detection reagent is added to the 96-well plate cells to detect the fluorescence intensity. Select the cell line that has fluorescence expression when not treated and the fluorescence light is reduced after inhibition, and further culture. The Colo205-LUC-TCF / LEF-M1 cell line is one of the cell lines screened above. Its growth curve, cell morphology, and cell growth status are similar to those of the original Colo205 cells. The ratio of its fluorescent signal with and without inhibitor treatment is the largest among all cell lines, and the ratio can reach 4-5 times when inhibited at 4 hours, which is completely suitable for the subsequent screening of Wnt inhibitors.

[0341] Experimental Example 2: Detection of the inhibitory ability of compounds on the Colo205-LUC-TCF / LEF-M1 reporter cell line. The Colo205-LUC-TCF / LEF-M1 cell line is a reporter tool cell stably transfected with the pGL4.49-LUC2-TCF / LEF vector, and its β-catenin Wnt pathway is continuously activated. After the addition of the inhibitor, the Wnt pathway is inhibited, and the expression level of firefly luciferase regulated by the TCF / LEF cis-element on the vector decreases. After the subsequent addition of the detection substrate, the detected light signal decreases accordingly, thereby detecting the inhibitory effect of the compound.

[0342] Add 100uL of the compound with a maximum concentration of 20uM to each well of a 96-well cell culture plate, and dilute the compound concentration by 3 times. Then inoculate 10,000 colo205 cells stably transfected with the reporter gene and 100uL of culture medium into each well, and make corresponding positive and negative control wells at the same time. Place the cells in a 5% CO2 cell culture incubator and culture at 37°C for 4h. After 4 hours, remove the culture medium, add 100uL of the reagent (Promega) containing the corresponding firefly luciferase substrate to each well, and measure the activity of the luciferase reporter gene. Use SpectraMax to read the luminescence intensity in full wavelength mode. The light signal intensity of cells treated only with DMSO is the positive control, and the light signal intensity of the cell-free well is the negative control. Calculate the IC50 concentration of each compound. The Colo 205 reporter gene detection data are summarized in Table 1.

[0343] Table 1. IC of compounds for inhibition of Colo205-LUC-TCF / LEF reporter gene 50 value

[0344]

[0345] Experimental Example 3: Proliferation inhibition test of compounds on Wnt mutant cell lines (Colo205, H929, HepG2 and DU4475) and non-Wnt mutant cell lines (RKO)

[0346] The cell lines used in the experiment are Colo205, H929, HepG2 and DU4475 cell lines in which the Wnt pathway is continuously activated and whose proliferation is Wnt pathway-dependent; and the RKO cell line in which the Wnt pathway is not activated under normal circumstances and whose proliferation is not dependent on the Wnt pathway is used as a control cell line to determine whether the inhibitory effect of the compounds of the present invention on Wnt-dependent proliferation is caused by other non-specific toxicity.

[0347] Colo205, H929, DU4475, HepG2 and RKO cell lines cultured in their respective culture media were treated during the logarithmic growth phase, and the cells were collected and prepared into a uniform cell suspension of known concentration, and then the cell suspension was added to a 96-well cell culture plate so that each well contained 1000-4000 cells. Place in a 5% CO2 cell culture incubator and culture at 37°C for 20-24 hours. On the second day, a completely dissolved, 3-fold gradient dilution of the compound was added to each cell culture well so that the final maximum concentration in the cell culture well was 10uM, and the culture was continued for 96 hours. This experiment uses Promega's cell activity detection test for detection. The more cells proliferate, the stronger the final signal intensity. The detection instrument is SpectraMax, full wavelength mode. The wells with only DMSO added were used as positive control wells, and the wells without cells were used as negative control wells. The IC50 value of each compound for the proliferation inhibition of cells with sustained Wnt activation or proliferation dependence, as well as the IC50 value for the proliferation inhibition of cells with inactivated Wnt or proliferation-independent Wnt were calculated to evaluate the inhibitory effect of the compound on the Wnt pathway and the toxic effect on normal cells (Table 2).

[0348] Table 2. IC values ​​of compounds for inhibition of proliferation of Wnt mutant cell lines 50 value

[0349]

[0350]

[0351] ND = Not Tested

[0352] The above results show that the compounds of the present invention have significant inhibitory activity against mutant cell lines Colo205, DU4475, NCI-H929 and HepG2, but have basically no significant inhibitory activity against Hela and RKO cell lines, which indicates that the compounds of the present invention have significant Wnt-dependent proliferation inhibitory effect.

Claims

1. A compound, or a pharmaceutically acceptable salt, isotopic derivative, or stereoisomer thereof, wherein the compound has the following structure: 。 2. A pharmaceutical composition comprising the compound according to claim 1 or a pharmaceutically acceptable salt, isotopic derivative, stereoisomer thereof, and optionally a pharmaceutically acceptable carrier.

3. Use of the compound according to claim 1 or its pharmaceutically acceptable salt, isotopic derivative, stereoisomer or the pharmaceutical composition according to claim 2 in the preparation of a medicament for preventing and / or treating cancer, tumor, inflammatory disease, autoimmune disease or immune-mediated disease.

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