A wnt pathway inhibitor compound

Abstract

The present application relates to Wnt pathway inhibitor compounds of Formula (I) and pharmaceutical compositions comprising the same, and the use of the compounds of Formula (I) for the prevention and / or treatment of cancer, a tumor, an inflammatory disease, an autoimmune disease, or an immune-mediated disease.

Description

[0001] This application claims priority to Chinese Patent Application No. 202110847130.9, filed on July 26, 2021, entitled “A Wnt Pathway Inhibitor Compound,” the contents of which are incorporated herein by reference in its entirety. TECHNICAL FIELD

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

[0003] The Wnt / β-catenin signaling pathway is a conserved pathway in biological evolution. In normal somatic cells, β-catenin only plays a role in maintaining homotypic cell adhesion and preventing cell movement as a cytoskeletal protein at the cell membrane in complex with E-cadherin. 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 through the proteasome pathway, maintaining β-catenin in the cytoplasm at a low level. When the cell is stimulated by Wnt signal, Wnt protein binds to specific receptor Frizzled protein on the cell membrane, and the activated Frizzled receptor recruits intracellular Dishevelled protein to inhibit the degradation activity of the β-catenin degradation complex formed by GSK3β and other proteins, stabilizing the free state of β-catenin protein in the cytoplasm. The stable accumulation of β-catenin in the cytoplasm enters the nucleus and binds to the LEF / TCF transcription factor family to initiate 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 canonical signaling pathway and nuclear accumulation of β-catenin protein are widespread in colorectal cancer, and inhibition of Wnt signaling pathway activity can inhibit the proliferation of cancer such as colon cancer. Mutations in APC occur in more than 85% of colorectal cancers, and the mutated APC blocks the phosphorylation and degradation of β-catenin, inducing the occurrence of colorectal cancer. In addition, Axin mutation, β-catenin mutation itself can also cause intracellular aggregation of β-catenin, activating the Wnt / β-catenin pathway.

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

[0005] The present application provides a compound having Wnt pathway activity having the structure of Formula (I) or a pharmaceutically acceptable salt, isotope derivative, stereoisomer thereof:

[0006]

[0007] wherein:

[0008] --- represents the presence or absence of a single bond;

[0009] R1 represents C1-C6 alkyl, C3-C6 cycloalkyl, 3-6 membered heterocycloalkyl, C6-C 10 aryl, 5-10 membered heteroaryl, and said R1 can be optionally substituted with 0, 1, 2, 3 substituents selected from: hydrogen, halogen, OR a , nitro, (C1-C6)alkyl, (C1-C6)alkoxy, (C1-C6)haloalkyl, (C3-C6)cycloalkyl, halo(C3-C6)cycloalkyl, 3-6 membered heterocycloalkyl, halo 3-6 membered heterocycloalkyl, C6-C 10 aryl, 5-10 membered heteroaryl, cyano, SR a , halo(C1-C6)alkoxy, halo(C3-C6)cycloalkoxy, halo(C1-C6)alkylthio, (C3-C6)cycloalkyloxy, (C3-C6)cycloalkylthio, halo(C3-C6)cycloalkylthio;

[0010] X represents a covalent bond or -(CR a R a’ ) m -, -(CR a Ra’) m -O-(CR a R a’ ) n -, -(CR a R a’ ) m -N(R b )-(CR a R a’ ) n -, -(CR a R a’ ) m -S-(CRa R a’ ) n -、-(CR a R a’ ) m C(O)(CR a R a’ ) n -、-(CR a R a’ ) m S(O)2(CR a R a’ ) n -、-(CR a R a’ ) m C(O)N(R b )(CR a R a’ ) n -、-(CR a R a’ ) m S(O)2N(R b )(CR a R a’ ) n -、-(CR a R a’ ) m N(R b )C(O)(CR a R a’ ) n -、-(CR a R a’ ) m N(R b )S(O)2(CR a R a’ ) n -、-(CR a R a’ ) m OC(O)N(R b )(CR a R a’ ) n -、-(CR a R a’ ) m N(R b )C(O)O(CR a R a’ ) n -、-(CR a R a ’) m N(R b )C(O)N(R b’ )(CRa R a’ ) n -、-(CR a R a’ ) m N(R b )S(O)2N(R b’ )(CR a R a’ ) n -;

[0011] Cy represents C3-C5cycloalkyl or 4-5 membered cycloheteroalkyl, and it can optionally be substituted with 0, 1 or 2 substituents selected from the group consisting of hydrogen, halogen, -OR a , (C1-C6)alkyl, (C1-C6)haloalkyl, (C3-C6)cycloalkyl, cyano and hydroxy(C1-C6)alkyl;

[0012] R 2 represents hydrogen, (C1-C6)alkyl, (C1-C6)haloalkyl, (C3-C6)cycloalkyl or hydroxy(C1-C6)alkyl;

[0013] R 3 and R 3’ each independently represent hydrogen, halogen, OR a , (C1-C6)alkyl, (C3-C6)cycloalkyl, hydroxyC1-C6alkyl or (C1-C6)alkoxy(C1-C6)alkyl;

[0014] or R 3 and R 3’ together with the carbon atom to which they are attached form a 3-6 membered saturated or unsaturated ring which can optionally contain 1 or 2 heteroatoms selected from O, S and N, and which ring can optionally be substituted with 0, 1 or 2 substituents selected from halogen, hydroxy and C1-C6alkyl;

[0015] or R 2 , R 3 or R 2 , R 3’ together with the atoms to which they are attached form a 4-6 membered saturated or unsaturated ring which can optionally contain 1 or 2 heteroatoms selected from O, S and N, and which ring can optionally be substituted with 0, 1, 2 halogen, hydroxy, C1-C6alkyl;

[0016] R 4 and R 4’ each independently represent hydrogen, C1-C6alkyl, C3-C6cycloalkyl, hydroxyC1-C6alkyl, haloC1-C6alkyl, (C1-C6)alkoxy(C1-C6)alkyl;

[0017] or R 4 forms =0 together with R 4’ ;

[0018] R T and R T’ each independently represent hydrogen, C1-C6alkyl, C1-C6haloalkyl, hydroxyC1-C6alkyl, C3-C6cycloalkyl, halogen, OR a ;

[0019] or R T and R T’ together with the atoms to which they are attached form a 3-6 membered ring;

[0020] wherein, when the double line---represents the absence of a single bond, A represents (CR L R L’ ) p wherein R L and R L’ each independently represent hydrogen, C1-C6alkyl, C1-C6haloalkyl, hydroxyC1-C6alkyl, C3-C6cycloalkyl, halogen, OR a , or R L and R L’ together with the carbon atom to which they are attached form a 3-6 membered ring, which ring can optionally contain 0, 1 or 2 heteroatoms selected from O, S and N, and which ring can also optionally be substituted with 0, 1 or 2 substituents selected from halogen and hydroxy;

[0021] wherein, when the double line---represents the presence of a single bond, A represents CR H wherein R H represents hydrogen, C1-C6alkyl, C1-C6haloalkyl, hydroxyC1-C6alkyl, C3-C6cycloalkyl, halogen, OR a ;

[0022] wherein R a , R a’ , R b , R b’ each independently represent hydrogen or C1-C6alkyl;

[0023] wherein m, n, p each independently represent 0, 1 or 2.

[0024] In the above compound of formula (I) or a pharmaceutically acceptable salt, isotopic derivative, stereoisomer thereof, it is preferably a compound of formula (II) or a pharmaceutically acceptable salt, isotopic derivative, stereoisomer thereof,

[0025]

[0026] wherein R 1 , R 2 , R 3 , R 3’ , R 4 , R 4’ , X, Cy are as defined above.

[0027] Further, the present application also provides a compound having the following formula (III) and pharmaceutically acceptable salts, isotope derivatives, stereoisomers:

[0028]

[0029] wherein R 1 , R 2 , R 3 , R 3’ , R 4 , R 4’ , R T , R T’ , R L , R L’ , X, Cy are as defined above.

[0030] In some embodiments of the present application, wherein R 2 represents hydrogen, C1-C6 alkyl, hydroxy(C1-C6 alkyl) or C3-C6 cycloalkyl.

[0031] In some embodiments of the present application, wherein R 3 , R 3’ each independently represents hydrogen, C1-C6 alkyl, hydroxy(C1-C6 alkyl) or C3-C6 cycloalkyl.

[0032] In some embodiments of the present application, wherein R 4 , R 4’ each independently represents hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl, or R 4 and R 4’ together form =O.

[0033] In some embodiments of the present application, wherein Cy represents C3-C5 cycloalkyl or 4-5 membered cycloheteroalkyl; preferably, Cy represents

[0034] In some embodiments of the present application, X represents a covalent bond or -(CR a R a’ ) m -, -(CR a R a’ ) m -O-(CR a Ra’ ) n -、-(CR a R a’ ) m -N(R b )-(CR a R a’ ) n -、-(CR a R a’ ) m -S-(CR a R a’ ) n -、-(CR a R a’ ) m C(O)N(R b )(CR a R a’ ) n -、-(CR a R a’ ) m S(O)2N(R b )(CR a R a’ ) n -、-(CR a R a’ ) m N(R b )C(O)(CR a R a’ ) n -、-(CR a R a’ ) m N(R b )S(O)2(CR a R a’ ) n -、-(CR a R a’ ) m OC(O)N(R b )(CR a R a’ ) n -、-(CR a R a’ ) m N(R b )C(O)O(CR a R a’ ) n -、-(CR a R a ’) m N(R b )C(O)N(R b’ )(CRa R a’ ) n -、-(CR a R a’ ) m N(R b )S(O)2N(R b’ (CR) a R a’ ) n -

[0035] In some embodiments of the present invention, X represents -O-, -NR b -、-CR a R a’ -、-OCR a R a’ -、-CR a R a’ O-、-C(O)-、-C(O)NR b -、-NR b C(O)-、-NR b -C(O)-NR b -、-CR a R a’ -C(O)NR b -、-CR a R a’ -NR b C(O)-, -S-, -NR b S(O)2-、-SO2NR b -OC(O)NR b -、-S(O)2、-C(O)NR b -、-C(O)CR a R a’ -、-CR a R a’ C(O)NR b -、-NR b C(O)CR a R a’ -、-NR b C(O)O-; where R a R a’ R b Each can be used independently to represent hydrogen or C1-C6 alkyl groups.

[0036] In some embodiments of the present invention, X represents -(CR a R a’ ) m -O-(CR a R a’ ) n -, preferably -O-, -OCRa R a’ -、-CR a R a’ O-, more preferably -O- or -O-CH2-.

[0037] In some embodiments of the present invention, R T and R T’ It represents hydrogen.

[0038] In some embodiments of the present invention, R L and R L’ It represents hydrogen.

[0039] In some embodiments of the present invention, R H It represents hydrogen.

[0040] In some embodiments of the present invention, R 1 This refers to C3-C6 cycloalkyl, 3-6 membered heterocycloalkyl, and C6-C6 cycloalkyl groups substituted with 0, 1, 2, or 3 substituents. 10 aryl, 5-10-membered heteroaryl, wherein the substituent is selected from halogen, (C1-C6)alkyl, (C1-C6)haloalkyl, (C3-C6)cycloalkyl, halo(C3-C6)cycloalkyl, 3-6-membered heterocycloalkyl, halo3-6-membered heterocycloalkyl, C6-C 10 Aryl and 5-10 heteroaryl compounds.

[0041] In some embodiments of the present invention, R 1 Indicates C3-C6 cycloalkyl, 3-6 membered heterocycloalkyl, C6-C 10 Aryl, 5-10 quinone heteroaryl, and the R 1 It can be arbitrarily replaced by 0, 1, 2, or 3 substituents selected from: hydrogen, halogen, (C1-C6)alkyl, (C1-C6)haloalkyl, (C3-C6)cycloalkyl, and halo(C3-C6)cycloalkyl.

[0042] In some embodiments of the present invention, R 1 Selected from:

[0043]

[0044] In some embodiments of the present invention, R 1 Selected from:

[0045]

[0046] In some preferred aspects, the present invention provides a compound having the following structure:

[0047]

[0048]

[0049]

[0050]

[0051]

[0052]

[0053]

[0054] Further, the present application also provides a pharmaceutical composition comprising the compound of the present application or a pharmaceutically acceptable salt, isotope derivative or stereoisomer thereof.

[0055] Further, the present application also provides the use of the compound of the present application or a pharmaceutically acceptable salt, isotope derivative or stereoisomer thereof, or the pharmaceutical composition of the present application in the manufacture of a medicament for the prevention and / or treatment of cancer, tumor, inflammatory disease, autoimmune disease or immune-mediated disease. It is particularly noted that, herein, when referring to a "compound" of the structure of Formula (I) to Formula (III), it generally also encompasses stereoisomers, diastereomers, enantiomers, racemic mixtures and isotope derivatives thereof.

[0056] Accordingly, the present application provides a method for the prevention and / or treatment of cancer, tumor, inflammatory disease, autoimmune disease or immune-mediated disease, comprising administering to a subject the compound of the present application or a pharmaceutically acceptable salt, isotope derivative or stereoisomer thereof, or the pharmaceutical composition of the present application.

[0057] It is well known to those skilled in the art that a salt, solvate, hydrate of a compound is an alternative form of the compound, which can be converted into the compound under certain conditions, therefore, it is particularly noted that, herein, when referring to a compound of the structure of Formula (I) to Formula (III), it generally also encompasses a pharmaceutically acceptable salt thereof, and further encompasses a solvate and a hydrate thereof.

[0058] Similarly, herein, when referring to a compound, it generally also encompasses a prodrug, metabolite and nitroxide thereof.

[0059] The pharmaceutically acceptable salts of the present application can be formed using, for example, inorganic or organic acids, as follows: "Pharmaceutically acceptable salt" refers to salts of a compound of the present application which are within the scope of sound medical judgment, which are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit / risk ratio. The salts can be prepared in situ during the final isolation and purification of the compounds of the present application, or separately by reacting the free base or free acid with a suitable reagent, as outlined below. For example, the free base functionality can be reacted with the appropriate acid. Examples of pharmaceutically acceptable inorganic salts are salts of hydrochloric, hydrobromic, phosphoric, sulfuric and perchloric acids, or organic acids such as acetic, oxalic, maleic, tartaric, citric, succinic or malonic acids, or by using other methods known in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hernisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxyethanesulfonate, 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 salts, and the like. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, loweralkyl sulfonate and aryl sulfonate.

[0060] The pharmaceutically acceptable salts of the present application can be prepared by conventional methods, for example by dissolving the compound of the present application in an organic solvent which is miscible with water, such as acetone, methanol, ethanol and acetonitrile, adding thereto an excess of an aqueous solution of an organic or inorganic acid, so that the salt precipitates from the resulting mixture, removing the solvent and the remaining free acid therefrom, and isolating the precipitated salt.

[0061] The precursors or metabolites described in this invention can be precursors or metabolites known in the art, as long as they are metabolized and transformed in vivo to form compounds. For example, "prodrug" refers to those prodrugs of the compounds of this invention that, within a reasonable medical judgment, are suitable for contact with human and lower animal tissues without undue toxicity, irritation, allergic reactions, etc., and have a reasonable benefit / risk ratio and are effective for their intended use. The term "prodrug" refers to a compound that is rapidly transformed in vivo to produce the parent compound of the above formula, for example, through in vivo metabolism, or through N-demethylation of the compounds of this invention.

[0062] The term "solvate" as used in this invention refers to the physical association of the compound of this invention with one or more solvent molecules (organic or inorganic). This physical association includes hydrogen bonding. In some cases, such as when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid, the solvate can be separated. The solvent molecules in the solvate may be present in a regular and / or disordered arrangement. The solvate may contain stoichiometric or non-stoichiometric solvent molecules. "Solvate" encompasses both 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.

[0063] The "stereoisomerism" described in this invention is divided into conformational isomerism and configurational isomerism. Configurational isomerism can be further divided into cis-trans isomerism and optical isomerism. Conformational isomerism refers to the phenomenon where organic molecules with a certain configuration undergo different spatial arrangements of atoms or groups of atoms due to the rotation or twisting of carbon or carbon single bonds. Common examples include the structures of alkanes and cycloalkanes, such as the chair conformation and boat conformation in the cyclohexane structure. "Stereoisomers" refer to compounds of this invention containing one or more asymmetric centers, thus allowing them to exist as racemic mixtures and racemic mixtures, single enantiomers, diastereomer mixtures, and single diastereomers. The compounds of this invention have asymmetric centers, each of which produces two optical isomers. The scope of this invention includes all possible optical isomers and diastereomer mixtures, as well as pure or partially pure compounds. The compounds of this invention can exist as tautomers, which have different hydrogen bonding sites through one or more double bond shifts. For example, ketones and their enol forms are ketone-enol tautomers. All tautomers and mixtures thereof are included in the compounds of this invention. All enantiomers, diastereomers, racemates, mesomates, cis-trans isomers, tautomers, geometric isomers, epimers, and mixtures thereof of compounds of formulas (I) to (III) are included within the scope of this invention.

[0064] An "isotopically-labeled" compound of the application is defined as a compound of the application having one or more atoms replaced by an isotope of the atom naturally occurring in the molecule. Isotopically-labeled compounds of this application are useful in metabolic studies, as diagnostic tools, as probes in assays, and the like. In particular, an isotopically-labeled compound of the application can serve as an active depot for purposes of delivery of the drug into tissues in the body. An isotopically-labeled compound of the application can also exhibit differences in metabolic stability or clearance compared to a non-labeled compound. 2 H and 3 H; carbon isotopes: 11 C, 13 C and 14 C; chlorine isotopes: 35 Cl and 37 Cl; fluorine isotopes: 18 F; iodine isotopes: 123 I and 125 I; nitrogen isotopes: 13 N and 15 N; oxygen isotopes: 15 O, 17 O and 18 O and sulfur isotopes 35 S. These isotopically-labeled compounds are useful in metabolic studies, as diagnostic tools, as probes in assays, and the like. In particular, an isotopically-labeled compound of the application can serve as an active depot for purposes of delivery of the drug into tissues in the body. An isotopically-labeled compound of the application can also exhibit differences in metabolic stability or clearance compared to a non-labeled compound. 3 H and carbon 13 C, are preferred for their ease of preparation and detectability. Substitution of heavier isotopes such as 2 H) can afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life, reduced dosage requirements, and / or attenuated dosage side effects.

[0065] The present application also provides the use of a compound of the present application for the manufacture of a medicament for the prevention and / or treatment of cancer, a tumor, an inflammatory disease, an autoimmune disease, or an immune-mediated disease.

[0066] In addition, the present application provides a pharmaceutical composition for the prevention and / or treatment of cancer, a tumor, an inflammatory disease, an autoimmune disease, a neurodegenerative disease, an attention-related disease, or an immune-mediated disease, comprising a compound of the present application as an active ingredient. The pharmaceutical composition can optionally comprise a pharmaceutically acceptable carrier.

[0067] In addition, the present application provides a method for the prevention and / or treatment of cancer, a tumor, an inflammatory disease, an autoimmune disease, a neurodegenerative disease, an attention-related disease, or an immune-mediated disease, comprising administering a compound of the present application to a mammal in need thereof.

[0068] Representative examples of inflammatory diseases, autoimmune diseases, and immune-mediated diseases can include, but are not limited to, arthritis, rheumatoid arthritis, spondyloarthritides, gouty arthritis, osteoarthritis, juvenile arthritis, other arthritic conditions, lupus, systemic lupus erythematosus (SLE), skin-related diseases, psoriasis, eczema, dermatitis, allergic dermatitis, pain, lung diseases, lung inflammation, adult respiratory distress syndrome (ARDS), pulmonary sarcoidosis, chronic lung inflammatory 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, Sjogren's syndrome, autoimmune thyroid disease, urticaria (hives), 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 rhinosinusitis, leukemia, lymphoma, B-cell lymphoma, T-cell lymphoma, myeloma, acute lymphoblastic leukemia (ALL), chronic lymphoblastic leukemia (CLL), acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), hairy cell leukemia, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, myelodysplastic syndrome (MDS), myeloproliferative neoplasm (MPN), diffuse large B-cell lymphoma, and follicular lymphoma.

[0069] Representative examples of cancer or tumors can 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 cancer, hereditary nonpolyposis colorectal cancer, esophageal cancer, lip cancer, larynx cancer, hypopharynx cancer, tongue cancer, salivary gland cancer, stomach cancer, adenocarcinoma, medullary thyroid cancer, papillary thyroid cancer, kidney cancer, renal parenchymal cancer, ovarian cancer, cervical cancer, uterine body cancer, endometrial cancer, choriocarcinoma, pancreatic cancer, prostate cancer, testicular cancer, urological cancer, melanoma, brain tumors such as glioblastoma, astrocytoma, meningioma, medulloblastoma, and peripheral neuroectodermal tumor, Hodgkin's lymphoma, non-Hodgkin's lymphoma, Burkitt's lymphoma, acute lymphoblastic leukemia (ALL), chronic lymphoblastic leukemia (CLL), acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), adult T-cell leukemia lymphoma, diffuse large B-cell lymphoma (DLBCL), hepatocellular carcinoma, gallbladder cancer, bronchial cancer, small cell lung cancer, non-small cell lung cancer, multiple myeloma, basal cell carcinoma, teratocarcinoma, retinoblastoma, choroidal melanoma, seminoma, rhabdomyosarcoma, craniopharyngioma, osteosarcoma, chondrosarcoma, myosarcoma, liposarcoma, fibrosarcoma, Ewing's sarcoma, or plasmacytoma.

[0070] When the compound of the present application or a pharmaceutically acceptable salt thereof is administered in combination with another anticancer agent or immune checkpoint inhibitor for the treatment of cancer or tumor, the compound of the present application or a pharmaceutically acceptable salt thereof can provide an enhanced anticancer effect.

[0071] Representative examples of the anticancer agent for the treatment of cancer or tumor can include, but are not limited to, cell signal transduction inhibitors, chlorambucil, melphalan, cyclophosphamide, ifosfamide, busulfan, carmustine, lomustine, streptozocin, 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, ixabepilone, tamoxifen, flutamide, goserelin analogs, megestrol, prednisone, dexamethasone, methylprednisolone, thalidomide, interferon alpha, leucovorin, sirolimus, sirolimus lipidate, everolimus, afatinib, alisertib, amuvatinib, apatinib, axitinib, bortezomib, bosutinib, brivanib, cabozantinib, cediranib, crenolanib, crotetuzumab, dabrafenib, dacotuzumab, danusertib, dasatinib, dovitinib, erlotinib, foretinib, ganetespib, gefitinib, ibrutinib, icotinib, imatinib, iniparib, lapatinib, lenvatinib, linifanib, linsitinib, masitinib, momelotinib, motesanib, neratinib, nilotinib, niraparib, oprozomib, olaparib, pazopanib, pictilisib, ponatinib, quizartinib, regorafenib, rigosertib, rucaparib, ruxolitinib, seribantumab, sorafenib, sunitinib, tivantinib, tivozanib, tofacitinib, trametinib, vandetanib, velpatasarin, vemurafenib, vismodegib, volasertib, alemtuzumab, bevacizumab, brentuximab vedotin, catumaxomab, cetuximab, denosumab, gemtuzumab ozogamicin, ipilimumab, nimotuzumab, ofatumumab, panitumumab, rituximab, tositumomab, trastuzumab, PI3K inhibitors, CSF1R inhibitors, A2A and / or A2B receptor antagonists, IDO inhibitors, anti-PD-1 antibodies, anti-PD-L1 antibodies, LAG3 antibodies, TIM-3 antibodies, and anti-CTLA-4 antibodies, or any combination thereof.

[0072] When the compound of the present application or a pharmaceutically acceptable salt thereof is administered in combination with another therapeutic agent for the treatment of inflammatory diseases, autoimmune diseases and immune-mediated diseases, the compound of the present application or a pharmaceutically acceptable salt thereof can provide an enhanced therapeutic effect.

[0073] Representative examples of therapeutic agents for the treatment of inflammatory diseases, autoimmune diseases and immune-mediated diseases can include, but are not limited to, steroidal drugs (e.g., prednisone, prednisolone, methylprednisolone, cortisone, hydrocortisone, betamethasone, dexamethasone, etc.), methotrexate, leflunomide, anti-TNFa agents (e.g., etanercept, infliximab, adalimumab, etc.), calcineurin inhibitors (e.g., tacrolimus, pimecrolimus, etc.), and antihistamines (e.g., diphenhydramine, hydroxyzine, loratadine, ebastine, ketotifen, cetirizine, levocetirizine, fexofenadine, etc.), and at least one or more therapeutic agents selected from among them can be included in the pharmaceutical composition of the present application.

[0074] The compound of the present application or a pharmaceutically acceptable salt thereof can be orally or parenterally administered as an active ingredient in an effective amount ranging from 0.1 to 2,000 mg / kg body weight / day, preferably 1 to 1,000 mg / kg body weight / day, in the case of a mammal including a human (body weight about 70 kg), and administered in a single or 4 divided doses per day, or in compliance with / incompliance with a predetermined time. The dose of the active ingredient can be adjusted according to various relevant factors (e.g., the condition of the subject to be treated, the type and severity of the disease, the rate of administration, and the physician's opinion). In some cases, an amount less than the above dose can be appropriate. An amount greater than the above dose can be used if it does not cause harmful side effects and can be administered in divided doses per day.

[0075] In addition, the present application also provides a method for preventing and / or treating a tumor, cancer, viral infection, organ transplant rejection, neurodegenerative disease, attention-related disease, or autoimmune disease, which comprises administering to a mammal in need thereof a compound of the present application or a pharmaceutical composition of the present application.

[0076] The pharmaceutical composition of the present application can be formulated into a dosage form for oral administration or parenteral administration (including intramuscular, intravenous, and subcutaneous routes, intratumoral injection) such as a tablet, granule, powder, capsule, syrup, emulsion, microemulsion, solution, or suspension according to any one of the conventional methods.

[0077] The pharmaceutical composition of the present application 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, a surfactant, a suspending agent, an emulsifying agent, and a diluent.

[0078] Examples of carriers employed in the pharmaceutical compositions of the present application for injection administration can be water, saline, dextrose solution, glucose-like solutions, alcohols, glycols, ethers (e.g., polyethylene glycol 400), oils, fatty acids, fatty acid esters, glycerides, surfactants, suspending agents, and emulsifiers.

[0079] Other features of the application will become apparent in the course of the detailed description which follows, given for purposes of illustration and not intended to be limiting of the application, the following examples use the methods disclosed herein to prepare, isolate and characterize.

[0080] The compounds of the present application can be prepared in a number of ways known to one skilled in the art of organic synthesis, using the methods described below, together with synthetic methods known in the art of synthetic organic chemistry, or by modifications thereof, as appropriate. Preferred methods include, but are not limited to, those described below. The reactions are performed in solvents appropriate to the reagents and materials employed and appropriate to the scale of the operation. The choice of reaction solvent and conditions will be influenced by many factors, for example, the functional groups present, the reagents used, and the scale of the reaction. The skilled worker will be able to determine appropriate reaction conditions. DETAILED DESCRIPTION

[0081] Definitions

[0082] The terms, as used in the present application, including the specification and claims, are defined as follows, if not otherwise indicated. The conventional methods of mass spectrometry, nuclear magnetic resonance, HPLC, protein chemistry, biochemistry, recombinant DNA techniques and pharmacology are used, if not otherwise indicated. In the present application, "or" or "and" means "and / or" if not otherwise indicated.

[0083] In the specification and claims, a given chemical formula or name shall cover all its stereoisomers and optical isomers and the racemates in which the above isomers exist. Unless otherwise specified, 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 the above stable isomers are covered by the present invention. The present invention describes the 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 forms. All methods for preparing the compounds of the present invention and the intermediates prepared therein are considered part of the present invention. When preparing enantiomeric or diastereomeric products, they can be separated by conventional methods (such as by chromatography or fractional crystallization). Depending on the method conditions, the end products of the present invention are obtained in free (neutral) or salt forms. Both the free forms and salts of these end products are within the scope of the present invention. If necessary, one form of the compound can be converted into another form. The free base or acid can be converted into a salt; the salt can be converted into a free compound or another salt; the mixture of isomeric compounds of the present invention can be separated into individual isomers. The compounds of the present invention, their free forms and salts can exist in multiple tautomeric forms, in which a hydrogen atom is transposed to other parts of the molecule and the chemical bonds between the atoms of the molecule are rearranged accordingly. It should be understood that all tautomeric forms that may exist are included in the present invention.

[0084] Unless otherwise defined, the definitions of the substituents of the present invention are independent of each other rather than interrelated. For example (by way of illustration and not limitation), in one aspect, for the substituent R a (or R a ’), it is independent in the definitions of different substituents. Specifically, for R a (or R a ’) when choosing one definition in one substituent, it does not mean that this R a [[ID=​​​​​​​​​​​​​​​​​​​​a When '), these R a (or R) a ') are also independent. For example, in the substituent -(CR a R a’ ) m -O-(CR a R a’ ) n In the case where m+n is greater than or equal to 2, there are m+n R values. a (or R) a Each ') is independent; they can have the same or different meanings.

[0085] Unless otherwise defined, when a substituent is labeled "optionally substituted," the substituent is selected from, for example, alkyl, cycloalkyl, aryl, heterocyclic, halogen, hydroxyl, alkoxy, oxo, alkanoyl, aryloxy, alkanoyloxy, amino, alkylamino, arylamino, arylalkylamino, disubstituted amino (where the two amino substituents are selected from alkyl, aryl, or arylalkyl), alkanoylamino, arylanoylamino, arylalkylamino, substituted alkanoylamino, substituted arylamino, substituted arylalkylamino, thio, alkylthio, arylthio, arylalkylthio, arylthiocarbonyl, arylalkylthiocarbonyl, Alkylsulfonyl, arylsulfonyl, arylalkylsulfonyl, aminosulfonyl (e.g., -SO2NH2), substituted sulfonylamino, nitro, cyano, carboxyl, carbamoyl (e.g., -CONH2), substituted carbamoyl (e.g., -CONHalkyl, -CONHaryl, -CONHarylalkyl or having two substituents selected from alkyl, aryl or arylalkyl on nitrogen), alkoxycarbonyl, aryl, substituted aryl, guanidine, heterocyclic groups (e.g., indolyl, imidazolyl, furanyl, thiophene, thiazolyl, pyrrolidinyl, pyridinyl, pyrimidinyl, pyrrolidinyl, piperidinyl, morpholinyl, piperazine, homopiperazine, etc.) and substituted heterocyclic groups.

[0086] As used herein, the term "alkyl" or "alkylene" is intended to include branched and straight-chain saturated aliphatic hydrocarbon groups having a specified number of carbon atoms. For example, "C1-C6 alkyl" refers to an alkyl group having 1 to 6 carbon atoms. Examples of alkyl groups 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 document, alkyl groups are preferably alkyl groups having 1 to 6, more preferably alkyl groups having 1 to 4 carbon atoms.

[0087] The term "alkenyl" denotes a straight or branched chain hydrocarbon group containing one or more double bonds and typically having a length of from 2 to 20 carbon atoms. For example, "C2-C6alkenyl" contains from two to six carbon atoms. Alkenyl groups include, but are not limited to, for example, ethenyl, propenyl, butenyl, 1 -methyl-2-buten-1 -yl, and the like. In the present context, alkenyl is preferably C2-C6alkenyl.

[0088] The term "alkynyl" denotes a straight or branched chain hydrocarbon group containing one or more triple bonds and typically having a length of from 2 to 20 carbon atoms. For example, "C2-C6alkynyl" contains from two to six carbon atoms. Representative alkynyl groups include, but are not limited to, for example, ethynyl, 1 -propynyl, 1 -butynyl, and the like. In the present context, alkynyl is preferably C2-C6alkynyl.

[0089] The term "alkoxy" or "alkyloxy" refers to -O-alkyl. "C1-C6alkoxy" (or alkyloxy) is intended to include C1, C2, C3, C4, C5, C6alkoxy groups. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propyloxy (for example, n- propoxy and isopropoxy), and t-butyloxy. In the present context, alkoxy is preferably alkoxy having from 1 to 6, more preferably having from 1 to 4 carbon atoms. Similarly, "alkylthio" or "thioalkyl" denotes an alkyl group as defined above attached to the rest of the molecule by a sulfur bridge; examples are methyl-S- and ethyl-S-.

[0090] The term "carbonyl" refers to the organic functional group (C=0) connected by a double bond to two atoms, one of which is carbon.

[0091] The term "aryl", alone or in combination with other terms, means a monocyclic, bicyclic or tricyclic ring system having from 5 to 12 ring members in which at least one ring is aromatic and wherein each ring in the system contains from 3 to 7 ring members. In certain embodiments of the application, "aryl" means 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" means an alkyl residue attached to an aryl ring, non-limiting examples of which include benzyl, phenethyl, and the like. A fused aryl group can be attached to another group at a suitable position on either the cycloalkyl ring or the aromatic ring. The dashed line drawn from the ring system indicates that the bond can be attached to any suitable ring atom.

[0092] The term "cycloalkyl" refers to monocyclic or bicyclic cyclic alkyl groups. Monocyclic cyclic alkyl groups refer to C3-C8cyclic alkyl groups, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and norbornyl. Branched cycloalkyl groups such as 1-methylcyclopropyl and 2-methylcyclopropyl are included in the definition of "cycloalkyl". Bicyclic cyclic alkyl groups include bridged, spirocyclic, or fused ring cyclic alkyl groups. Herein, cycloalkyl is preferably C3-C6cycloalkyl.

[0093] The term "cycloalkenyl" refers to monocyclic or bicyclic cyclic alkenyl groups. Monocyclic cyclic alkenyl groups refer to C3-C8cyclic alkenyl groups, including but not limited to cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, and norbornenyl. Branched cycloalkenyl groups such as 1-methylcyclopropenyl and 2-methylcyclopropenyl are included in the definition of "cycloalkenyl". Bicyclic cyclic alkenyl groups include bridged, spirocyclic, or fused ring cyclic alkenyl groups.

[0094] "Halo" or "halogen" includes fluorine, chlorine, bromine, and iodine. "Haloalkyl" is intended to include both branched and straight chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms 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 heptafluoropropyl. Examples of haloalkyl also include "fluoroalkyl" intended to include both branched and straight chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms substituted with one or more fluorine atoms.

[0095] "Haloalkoxy" or "haloalkyloxy" denotes an oxygen-bridged haloalkyl group as defined above having the specified number of carbon atoms. For example, "haloCi-C6alkoxy" is intended to include Ci, C2, C3, C4, C5, C6haloalkoxy groups. Examples of haloalkoxy include, but are not limited to, trifluoromethoxy, 2,2,2-trifluoroethoxy, and pentafluoroethoxy. Similarly, "haloalkylthio" or "thiohaloalkoxy" denotes a sulfur-bridged haloalkyl group as defined above having the specified number of carbon atoms; for example trifluoromethyl-S- and pentafluoroethyl-S-.

[0096] In the present disclosure, when referring to some substituent groups, C x1 -C x2In the present disclosure, the expression "C0-C8" indicates that the number of carbon atoms in the substituent group can be from x1 to x2. For example, C0-C8 indicates that the group contains 0, 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms, Ci-C8 indicates that the group contains 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms, C2-C8 indicates that the group contains 2, 3, 4, 5, 6, 7, or 8 carbon atoms, C3-C8 indicates that the group contains 3, 4, 5, 6, 7, or 8 carbon atoms, C4-C8 indicates that the group contains 4, 5, 6, 7, or 8 carbon atoms, C0-C6 indicates that the group contains 0, 1, 2, 3, 4, 5, or 6 carbon atoms, Ci-C6 indicates that the group contains 1, 2, 3, 4, 5, or 6 carbon atoms, C2-C6 indicates that the group contains 2, 3, 4, 5, or 6 carbon atoms, C3-C6 indicates that the group contains 3, 4, 5, or 6 carbon atoms.

[0097] In the present disclosure, the expression "3-12 membered ring" when referring to a cyclic group (e.g., aryl, heteroaryl, cycloalkyl, and heterocycloalkyl) indicates that the number of ring atoms in the group can be from x1 to x2. For example, the 3-12 membered cyclic group can be a 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 membered ring, which can have 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 ring atoms; 3-6 membered ring indicates that the cyclic group can be a 3, 4, 5, or 6 membered ring, which can have 3, 4, 5, or 6 ring atoms; 3-8 membered ring indicates that the cyclic group can be a 3, 4, 5, 6, 7, or 8 membered ring, which can have 3, 4, 5, 6, 7, or 8 ring atoms; 3-9 membered ring indicates that the cyclic group can be a 3, 4, 5, 6, 7, 8, or 9 membered ring, which can have 3, 4, 5, 6, 7, 8, or 9 ring atoms; 4-7 membered ring indicates that the cyclic group can be a 4, 5, 6, or 7 membered ring, which can have 4, 5, 6, or 7 ring atoms; 5-8 membered ring indicates that the cyclic group can be a 5, 6, 7, or 8 membered ring, which can have 5, 6, 7, or 8 ring atoms; 5-12 membered ring indicates that the cyclic group can be a 5, 6, 7, 8, 9, 10, 11, or 12 membered ring, which can have 5, 6, 7, 8, 9, 10, 11, or 12 ring atoms; 6-12 membered ring indicates that the cyclic group can be a 6, 7, 8, 9, 10, 11, or 12 membered ring, which can have 6, 7, 8, 9, 10, 11, or 12 ring atoms. The ring atoms can be carbon atoms or heteroatoms, e.g., heteroatoms selected from N, O, and S. When the ring is a heterocycle, the heterocycle can contain 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more ring heteroatoms, e.g., heteroatoms selected from N, O, and S.

[0098] In the present disclosure, one or more halogen can each independently be selected from fluorine, chlorine, bromine, and iodine.

[0099] The term "heteroaryl" means a stable 3-, 4-, 5-, 6-, or 7-membered aromatic monocyclic or aromatic bicyclic or 7-, 8-, 9-, 10-, 11-, 12-membered polyaromatic heterocyclic ring which is fully or partially unsaturated and which contains carbon atoms and 1, 2, 3, or 4 heteroatoms independently selected from N, O, and S; and includes any of the following polycyclic groups wherein any of the heterocyclic rings defined above are fused to a benzene ring. The nitrogen and sulfur heteroatoms can optionally be oxidized. The nitrogen atoms are substituted or unsubstituted (i.e., N or NR, where R is H or, if defined, another substituent). The heterocyclic rings can be attached to their side groups at any heteroatom or carbon atom that results in a stable structure. The heterocyclyl groups described herein can be substituted on a carbon or a nitrogen atom if the resulting compound is stable. The nitrogens in the heterocyclic ring can optionally be quaternized. Preferably, when the total number of S and O atoms in the heterocyclic ring exceeds one, then these heteroatoms are not adjacent to one another. Preferably, the total number of S and O atoms in the heterocyclic ring is not more than one. When the term "heterocycle" is used, it is intended to include heteroaryl.Examples of heteroaryl groups include, but are not limited to, acridinyl, azetidinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzoxazolinyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, imidazopyridinyl, indoleninyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isatinoyl, isochromanyl, isochromenyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isothiazolopyridinyl, isoxazolyl, isoxazolopyridinyl, methylenedioxyphenyl, morpholinyl, naphthizidinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolopyridinyl, oxazolidinyl, perimidinyl, oxindolyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, piperidonyl, 4-piperidonyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolopyridinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2-pyrrolidonyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrazolyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, 6H-1,2,5-thiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienopyridinyl, thienothiazolyl, thienooxazolyl, thienoimidazolyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, and xanthenyl, quinolinyl, isoquinolinyl, phthalazinyl, quinazolinyl, indolyl, isoindolyl, indolinyl, 1H-indazolyl, benzimidazolyl, 1,2,3,4-tetrahydroquinolinyl, 1,2,3,4-tetrahydroisoquinolinyl, 5,6,7,8-tetrahydro-quinolinyl, 2,3-dihydro-benzofuranyl, chromanyl, 1,2,3,4-tetrahydro- quinoxalinyl and 1,2,3,4-tetrahydro-quinazolinyl.The term "heteroaryl" can also include biaryl structures formed from the above defined "aryl" groups with monocyclic "heteroaryl" groups, such as, but not limited to, "-phenylbipyridyl-", "-phenylbipyrimidyl-", "-pyridylbiphenyl-", "-pyridylbipyrimidyl-", "-pyrimidylbiphenyl-"; wherein the present application also includes fused ring and spiro compounds containing, for example, the above heterocycles.

[0100] The term "heterocycloalkyl" as used herein refers to a monocyclic heterocycloalkyl ring system, or to a bicyclic heterocycloalkyl ring system, and also includes spiro or bridged heterocycloalkyl groups. Monocyclic heterocycloalkyl refers to a 3-8 membered, saturated or unsaturated, but not aromatic, ring system containing at least one heteroatom selected from O, N, S and P. Bicyclic heterocycloalkyl ring systems refer to a heterocycloalkyl ring fused to a phenyl, or a cycloalkyl, or a cycloalkenyl, or a heterocycloalkyl, or a heteroaryl ring.

[0101] The term "bridged cycloalkyl" as used herein refers to polycyclic compounds sharing two or more carbon atoms. It can be divided into bicyclic bridged cycloalkanes and polycyclic bridged cycloalkanes. The former consists of two alicyclic rings sharing two or more carbon atoms; the latter consists of three or more rings of bridged cycloalkanes.

[0102] The term "spirocycloalkyl" as used herein refers to polycyclic hydrocarbons sharing one carbon atom (called the spiro atom) between single rings.

[0103] The term "bridged heterocyclyl" as used herein refers to polycyclic compounds sharing two or more carbon atoms, at least one of which is a heteroatom selected from O, N and S. It can be divided into bicyclic bridged heterocycles and polycyclic bridged heterocycles.

[0104] The term "spiroheterocyclyl" as used herein refers to polycyclic hydrocarbons sharing one carbon atom (called the spiro atom) between single rings, at least one of which is a heteroatom selected from O, N and S.

[0105] The term "substituted" as used herein means that at least one hydrogen atom has been replaced by a non-hydrogen group, provided that a stable compound results. A ring double bond as used herein is a double bond between two adjacent ring atoms (e.g., C=C, C=N or N=N).

[0106] In cases wherein there are nitrogen atoms (e.g., amines) on the compounds of the present application, these nitrogen atoms can be converted to N-oxides by treatment with an oxidizing agent (e.g., mCPBA and / or hydrogen peroxide) to afford additional compounds of the present application. Thus, shown and claimed nitrogen atoms are considered to cover both the shown nitrogen and its N-oxide to afford derivatives of the present application.

[0107] When any variable occurs more than one time in any constituent or formula, its definition in each occurrence is independent of its definition in every other case. Thus, for example, if a group is shown to be substituted with 0 to 3 R groups, said group can optionally be substituted with up to three R groups, and at each occurrence R is independently selected from the definition of R. Also, combinations of substituents and / or variables are permissible only if such combinations result in stable compounds.

[0108] The term "patient" as used herein refers to an organism to be treated by the methods of the present application. Such organisms preferably include, but are not limited to, mammals (e.g., murines, simians, monkeys, horses, cows, pigs, dogs, cats, etc.) and most preferably refer to humans.

[0109] The term "effective amount" as used herein means that amount of a drug or pharmaceutical agent (i.e., a compound of the present application), which will elicit the biological or medical response of a tissue, system, animal, or human that is being sought, for instance, by a researcher or clinician. Furthermore, the term "therapeutically effective amount" means an amount of a compound effective to improve the treatment, cure, prevent, or reduce the severity of a disease, condition, or adverse effect, or to enhance normal physiological function, when administered to a subject in need thereof. An effective amount can be given in one or more administrations, applications, or dosages and is not intended to be limited to a particular formulation or route of administration. The term also includes within its scope amounts effective to enhance normal physiological function.

[0110] The term "treatment" as used herein includes any effect that relieves, reduces, modulates, ameliorates, or eliminates a condition, disease, disorder, etc., or its symptoms.

[0111] The term "pharmaceutically acceptable" is used herein to refer to those compounds, materials, 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 problem or complication commensurate with a reasonable benefit / risk ratio.

[0112] The phrase "pharmaceutically acceptable carrier" as used herein means a pharmaceutically-acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, manufacturing aid (e.g., lubricant, talc, magnesium stearate, calcium stearate or zinc stearate, or steric acid), or solvent encapsulating material, involved in carrying or transporting the subject compound from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceutically-acceptable carriers include: (1) sugars, such as glucose or beta-lactose;

[0113] The term "pharmaceutical composition" means a composition comprising a compound of the present application in combination with at least one other pharmaceutically acceptable carrier. "Pharmaceutically acceptable carrier" means a medium generally accepted in the art for the delivery of a biologically active agent to an animal, particularly a mammal, and includes, i.e., an adjuvant, excipient, or vehicle such as diluent, preservative, filler, flow regulator, disintegrating agent, wetting agent, emulsifying agent, suspending agent, sweetening agent, flavoring agent, perfuming agent, antibacterial agent, antifungal agent, lubricating agent, and dispersing agent, depending on the nature of the mode of administration and dosage form.

[0114] Specific Pharmaceutical and Medical Terms

[0115] The term "acceptable", as used herein, means no undue harmful effect on the general health of the subject of treatment of the active ingredients of the prescription.

[0116] The term "cancer", as used herein, means an uncontrolled abnormal growth of cells and, under certain conditions, the ability to metastasize (spread). This type of cancer includes, but is not limited to, solid tumors (e.g., bladder, bowel, brain, breast, uterine, cardiac, kidney, lung, lymphatic tissue (lymphoma), ovarian, pancreatic or other endocrine organ (e.g., thyroid), prostate, skin (melanoma), or blood tumors (e.g., non-leukemic leukemia).

[0117] The term "co-administration" or its grammatical equivalents, as used herein, means the administration of two or more selected therapeutic agents to a single patient at the same time by the same or different routes of administration.

[0118] The term "enhance" or "enhancing", as used herein, means an intended result that can be an increase or prolongation in potency or duration. Thus, in the context of enhancing the therapeutic effect of a drug, the term "enhancing" means the ability of a drug to increase or prolong the potency or duration of action in a system. "Enhancing value", as used herein, means the ability to maximize the enhancement of another therapeutic agent in an ideal system.

[0119] The term "immune disease" means a disease or condition resulting from an adverse or deleterious reaction to an endogenous or exogenous antigen. The result is usually a dysfunction of cells, or destruction and dysfunction as a result, or destruction of an organ or tissue that can produce the immune condition.

[0120] The terms "kit" and "product package" are synonymous.

[0121] The term "subject" or "patient" includes mammals and non-mammals. Mammals include, but are not limited to, mammals: humans, non-human primates such as chimpanzees, and other apes and monkey species; farm animals such as cattle, horses, sheep, goats, swine; domestic animals such as rabbits, dogs, and house cats; laboratory animals such as rats, mice, and guinea pigs; and the like. Non-mammalian animals include, but are not limited to, birds, fish, and the like. In a preferred aspect, the mammal is a human.

[0122] The terms "treatment," "treatment regime," or "therapy" as used herein, include alleviating, inhibiting or ameliorating a disease symptom or condition; inhibiting the onset of a complication; ameliorating or preventing a latent metabolic syndrome; inhibiting the development of a disease or condition, such as controlling the progression of a disease or condition; abating a disease or condition; causing regression of a disease or condition; relieving a complication caused by a disease or condition, or preventing and / or treating an indication caused by a disease or condition.

[0123] As used herein, an improvement in a disease, condition, or condition, refers to an improvement in the disease, condition, or condition, particularly an improvement in the severity thereof, a delay in onset, a slowing of progression, or a decrease in the duration thereof, whether fixed or contingent, continuous or intermittent, as a result of or in connection with the administration.

[0124] Routes of administration

[0125] Suitable routes of administration include, but are not limited to, oral, intravenous, rectal, aerosol, parenteral, ocular, pulmonary, transdermal, vaginal, aural, nasal, and topical. In addition, parenteral administration includes, by way of example only, intramuscular, subcutaneous, intravenous, intramedullary injections, intraventricular injection, intraperitoneal injection, intralymphatic injection, and intranasal injection.

[0126] In one aspect, the compounds described herein are administered in a manner that is local rather than systemic. In a particular embodiment, the long-acting formulation is administered by implantation (e.g., subcutaneously or intramuscularly) or by intramuscular injection. In addition, in another embodiment, the drug is administered by a targeted drug delivery system. For example, a liposome encapsulated with an organ-specific antibody. In this embodiment, the liposome is selectively directed to a particular organ and is taken up.

[0127] Pharmaceutical compositions and dosages

[0128] The present application also provides pharmaceutical compositions comprising a therapeutically effective amount of one or more compounds of the present application formulated together with one or more pharmaceutically acceptable carriers (additives) and / or diluents and, optionally, one or more other therapeutic agents as described above. The compounds of the present application can be administered by any suitable means, for any of the above indications, e.g., orally, such as tablets, capsules, powders, granules, elixirs, tinctures, suspensions including nanosuspensions, microsuspensions, spray-dried dispersions, syrups, and emulsions; sublingually; buccally; parenterally, such as by subcutaneous, intravenous, intramuscular, or intrasternal injection or infusion techniques (e.g., as sterile injectable aqueous or nonaqueous solutions or suspensions); nasally, including to the nasal membranes, such as by inhalable sprays; topically, such as in creams or ointments; or rectally such as in suppositories; or intratumorally. They can be administered alone, but generally are given in conjunction with pharmaceutical carriers selected on the basis of the chosen route of administration and standard pharmaceutical practice.

[0129] Pharmaceutical carriers are routinely selected based on a variety of factors, including, but not limited to: the type and nature of the active agent being formulated; the subject to which the active agent-containing composition is to be administered; the intended route of administration of the composition; and the therapeutic indication being targeted. Pharmaceutical carriers include aqueous and nonaqueous liquid media and various solid and semi-solid pharmaceutical carriers.

[0130] The above carriers can include a wide variety of different ingredients and additives in addition to the active agent, including other ingredients in the formulation for a variety of reasons known to those of skill in the art, such as stabilizers, binders, etc. Descriptions of suitable pharmaceutical carriers and factors involved in selection of carriers can be found in multiple readily available sources, such as Allen L.V. Jr. et al. Remington: The Science and Practice of Pharmacy (2 Volumes), 22nd Edition (2012), Pharmaceutical Press.

[0131] The dosage regimen for compounds of the present application will, of course, depend on known factors, such as the pharmacodynamic characteristics of the particular agent and its mode and route of administration; the species, age, sex, health, medical condition, and weight of the recipient; the nature and extent of the symptoms; the kind of concurrent treatment(s) and the frequency and route of administration, the renal and hepatic function of the patient, and the dosage should the desired effect be unsatisfactory. Under normal conditions, oral daily dosage levels of each active ingredient for the desired effects will be from about 0.001 mg / day to about 10-5000 mg / day, preferably from about 0.01 mg / day to about 1000 mg / day, and most preferably from about 0.1 mg / day to about 250 mg / day, when used for the indicated effects. Intravenous dosage levels during constant rate infusion will most preferably be from about 0.01 mg / kg / min to about 10 mg / kg / min. Compounds of the present application can be administered in a single daily dose, or the total daily dosage can be administered in divided doses of two, three or four times daily.

[0132] The compounds are generally administered in the form of a pharmaceutical composition in admixture with a suitable pharmaceutical diluent, excipient, or carrier (collectively referred to as a pharmaceutical carrier) appropriately selected with respect to the intended form of administration, e.g. oral tablets, capsules, elixirs, and syrups for oral administration, and liquid oral dosage forms for oral administration.

[0133] Dosage forms (pharmaceutical compositions) suitable for administration include about 1 milligram to about 2000 milligrams of active ingredient per dosage unit.

[0134] A typical capsule for oral administration contains at least one compound of the present application (250 mg), lactose (75 mg) and magnesium stearate (15 mg). The mixture is passed through a 60 mesh sieve and packed into a No. 1 gelatin capsule.

[0135] A typical injectable formulation can be prepared as follows: at least one compound of the present application (250 mg) is placed in a vial, lyophilized from sterile water, and the vial sealed. For use, the contents of the vial is reconstituted with 2 mL of sterile water to give an injectable formulation.

[0136] The scope of the present application includes pharmaceutical compositions comprising, as an active ingredient, a therapeutically effective amount of at least one compound of the present application, alone or in combination with a pharmaceutical carrier. Optionally, the compounds of the present application can be used in combination with other compounds of the present application or with one or more other therapeutic agents (e.g., anti-cancer agents or other pharmaceutically active substances).

[0137] Regardless of the route of administration selected, the compounds of the present application, which can be used in a suitable hydrated form, and / or the pharmaceutical compositions of the present application, are formulated into pharmaceutically-acceptable dosage forms by conventional methods known to those of skill in the art.

[0138] Actual dosage levels of the active ingredients in the pharmaceutical compositions of this application can be varied so as to obtain an amount of the active compound(s) that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

[0139] The selected dosage level will depend on a variety of factors including the activity of the particular compound of the present application employed, or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion of the particular compound being employed, the rate and extent of absorption, the duration of the treatment, other drugs, compounds and / or materials used in combination with the particular compound employed, the age, sex, weight, condition, general health and pre-existing medical conditions of the patient being treated, and like factors well known in the medical arts.

[0140] A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the compounds of the application employed in the pharmaceutical composition at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. In general, a suitable daily dose of a compound of the application will be in the range from about 0.01 to about 50 mg / kg body weight per day. If desired, the effective daily dose can be divided into multiple doses for purposes of administration, optionally in unit dosage forms. In certain aspects of the application, dosing is once daily.

[0141] While it is possible for a compound of the present application to be administered alone, it is preferable to present the compound as a pharmaceutical formulation (composition).

[0142] Kits / Products Packages are also described herein for use in the treatment of the indications described above. These kits can comprise a carrier, package, or container housing the components. Suitable containers include bottles, tubes, syringes, and test tubes, among others. The containers can be formed from a variety of materials such as glass or plastic.

[0143] For example, the container(s) can hold one or more of the compounds described herein, which can be present as a pharmaceutical composition, or as a mixture of compounds with other ingredients described herein. The container(s) can have a sterile access port (for example the container(s) can be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). Such kits can include one or more compounds, and instructions for use in the methods described herein, a label, or a package insert.

[0144] A typical kit can include one or more containers, each containing one or more of the materials (e.g., reagents, which can also be provided as concentrated stocks, and / or devices) needed for performance of the compound use, as is appropriate and useful for commercial sale and use of the compound. Such materials include, but are not limited to, buffers, diluents, filters, needles, syringes, delivery devices, packs, containers, bottles, and / or tubes, with or without an internal package, and with or without an instruction sheet attached or associated with the container. All of the instructions can be included.

[0145] Labels suitable for use in conjunction with the containers and reagents of the application include those that are affixed to the container, to a label associated with the container, or to a document contained in the container. Labels appear on containers, e.g., by being adhered to or imprinted on the container, or appear on a label associated with the container, e.g., by being adhered to or imprinted on the label. Labels can also appear on packaging or shipping containers, e.g., by being adhered to or imprinted on the packaging or shipping containers, or by being adhered to or imprinted on a document contained in the packaging or shipping containers, e.g., a product insert. A label can be used to indicate that the contents are associated with a particular therapeutic use. A label can also indicate directions for use of the contents, e.g., as described in the methods above.

[0146] All of the features described in this specification (including any accompanying claims, abstract and drawings), and / or all of the steps of any method or process described in this specification, can be combined in any combination, unless the context explicitly indicates that some features, processes or steps cannot be combined in that manner.

[0147] Any of the features of the application described above, or of the embodiments described above, can be combined in any combination. All of the features disclosed in this specification (and any accompanying claims, abstract and drawings) can be replaced by alternative features serving the same, equivalent or similar purpose, unless expressly stated otherwise. Thus, unless expressly stated otherwise, the features disclosed are only examples of a generic series of equivalent or similar features.

[0148] The application is further defined by reference to the following examples. It will be appreciated that these examples are only by way of illustration and are not intended to limit the scope of the application. Unless otherwise indicated, the methods of the following examples were carried out in accordance with conventional procedures or as otherwise described. Unless otherwise indicated, all percentages, ratios, proportions, or parts, are by weight.

[0149] The units of weight and volume percent in the present application are well known to those skilled in the art, e.g., refer to the weight (g) of solute in 100 mL of solution. Unless otherwise defined, all other terms of art used herein have the same meaning as commonly understood by one of ordinary skill in the art in the field of the application. In addition, any method or material similar or equivalent to those described herein can be used in the practice of the present application. The present description and examples are only exemplary and illustrative of the application.

[0150] Examples

[0151] General procedure

[0152] The starting materials and reagents used in the present application are known products when the preparation route is not included, and can be synthesized according to the methods known in the art, or can be obtained by purchasing commercially available products. The commercially available reagents used do not need to be further purified.

[0153] Room temperature refers to 20-30℃.

[0154] Unless otherwise specified in the reaction examples, the reactions are carried out under a nitrogen atmosphere. The nitrogen atmosphere refers to that the reaction bottle is connected to a nitrogen balloon of about 1 L.

[0155] The hydrogenation reaction is usually vacuumed and filled with hydrogen, and the operation is repeated 3 times. The hydrogen atmosphere refers to that the reaction bottle is connected to a hydrogen balloon of about 1 L.

[0156] Microwave reaction uses Initiator + microwave reactor.

[0157] The structure of the compound of the present application is determined by nuclear magnetic resonance (NMR) and mass spectrometry (MS). The NMR shift (δ) is given in units of 10 -6 (ppm). The NMR is measured by a (Bruker Ascend TM 500 type) nuclear magnetic instrument, and the measuring solvent is deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD), and the internal standard is tetramethylsilane (TMS). The following abbreviations are used for the multiplicity of NMR signals: s = singlet, brs = broad singlet, d = doublet, t = triplet, m = multiplet. The coupling constant is listed as J value, measured in Hz.

[0158] The measurement of LC-MS uses Thermo liquid chromatograph-mass spectrometer (UltiMate 3000+MSQ PLUS). The measurement of HPLC uses Thermo high-pressure liquid chromatograph (UltiMate 3000). The reverse phase preparation chromatography uses Thermo (UltiMate 3000) reverse phase preparation chromatograph. The flash column chromatography uses Ajinomoto (FS-9200T) automatic column machine, and the silica gel pre-packed column uses Sanso 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 the thin layer chromatography separation and purification product is 0.4mm-0.5mm.

[0159] Example 1

[0160] (S)-4,5-Dimethyl-2-((trans-3-(3,4,5-trifluorophenoxy)cyclobutyl)amino)-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0161]

[0162] Compound 1 was prepared from the following steps:

[0163]

[0164] First step: cis-3-BOC-aminocyclobutanol la (250 mg, 1.34 mmol), methyl sulfonic anhydride (465 mg, 2.67 mmol) and N,N-diisopropylethylamine (517 mg, 4.01 mmol) were dissolved in dichloromethane (2 mL) and stirred at room temperature overnight. The reaction was monitored by TLC and the reaction was diluted with dichloromethane and washed with water and saturated brine successively. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give yellow solid lb (300 mg, yield 84%). 1 H NMR (500 MHz, DMSO-d6) δ 7.23 (d, J = 8.3 Hz, 1H), 4.69-4.64 (m, 1H), 3.63-3.60 (m, 1H), 3.13 (s, 3H), 2.70-2.62 (m, 2H), 2.16-2.09 (m, 2H), 1.37 (s, 9H).

[0165] Second step: Compound lb (300 mg, 1.13 mmol), compound lc (251 mg, 1.70 mmol) and cesium carbonate (737 mg, 2.26 mmol) were dissolved in N,N-dimethylformamide (2 mL) and stirred at 80 °C overnight. The reaction was monitored by LCMS and the reaction was diluted with ethyl acetate and washed with water and saturated brine successively. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to give white solid Id (280 mg, yield 78%). ESI-MS (m / z): 318.6 [M+H] + .

[0166] Third step: Compound Id (280 mg, 882 umol) was dissolved in dichloromethane (2 mL) and hydrochloric acid 1,4-dioxane solution (4 M, 1.10 mL) was added dropwise and stirred at room temperature overnight. The reaction was monitored by LCMS and the reaction was concentrated to give white solid le (170 mg, yield 75%). ESI-MS (m / z): 218.4 [M+H] + .

[0167] Fourth Step: Dissolve 2,4-dichloropyrido[3,2-d]pyrimidine 1f (1.7 g, 8.50 mmol) and (S)-methyl 2-(methylamino)propanoate hydrochloride 1g (1.70 g, 11.05 mmol) in tetrahydrofuran (40 mL), add triethylamine (2.58 g, 25.50 mmol, 3.53 mL), stir at room temperature overnight. Monitor the reaction by LCMS until completion. Concentrate the reaction mixture and purify the residue by silica gel column chromatography to give yellow oil 1h (1.1 g, yield 46%). ESI-MS (m / z): 281.2 [M+H] + .

[0168] Fifth Step: Dissolve compound 1h (1.1 g, 3.92 mmol) in tetrahydrofuran (20 mL), add hydrochloric acid aqueous solution (6 N, 0.65 mL) and platinum dioxide (88 mg, 0.39 mmol), replace the reaction system with a hydrogen balloon, stir at room temperature under the pressure of the hydrogen balloon for 48 hours. Monitor the reaction by LCMS until completion. Dilute the reaction mixture with methanol, filter, concentrate the filtrate and purify by silica gel column chromatography to give white solid 1i (900 mg, yield 90%). ESI-MS (m / z): 253.2 [M+H] + .

[0169] Sixth Step: Dissolve compound 1i (50 mg, 197 umol), compound 1e (65 mg, 257 umol) and p-toluenesulfonic acid monohydrate (3.7 mg, 19 umol) in n-butanol (2 mL), react at 160 °C under microwave for 2 hours. Monitor the reaction by LCMS until completion. Purify the reaction mixture by reverse phase preparative HPLC to give white solid 1 (13 mg, yield 15%). ESI-MS (m / z): 434.3 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) δ 6.90-6.82 (m, 3H), 4.86-4.81 (m, 1H), 4.42-4.36 (m, 1H), 4.12 (q, J = 6.8 Hz, 1H), 4.05-4.00 (m, 1H), 3.30-3.24 (m, 1H), 2.94 (s, 3H), 2.55-2.52 (m, 2H), 2.48-2.38 (m, 2H), 2.36-2.28 (m, 2H), 1.97-1.88 (m, 1H), 1.85-1.76 (m, 1H), 1.23 (d, J = 6.8 Hz, 3H).

[0170] Example 2

[0171] (S)-4,5-dimethyl-2-((trans-3-((6-(trifluoromethyl)pyridin-3-yl)oxy)cyclobutyl)amino)- 4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0172]

[0173] Compound 2 was prepared from the following steps:

[0174]

[0175] First step: Compound 1b (500 mg, 1.88 mmol), compound 2a (461 mg, 2.83 mmol) and cesium carbonate (1.23 g, 3.77 mmol) were dissolved in N,N-dimethylformamide (2 mL) and stirred at 80 °C overnight. LCMS monitored the reaction was completed, the reaction was diluted with ethyl acetate, washed with water and saturated brine successively, the organic phase was dried over anhydrous sodium sulfate, the reaction was concentrated, the residue was purified by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to give white solid 2b (500 mg, yield 79%). ESI-MS (m / z): 333.3 [M+H] + .

[0176] Second step: Compound 2b (500 mg, 1.50 mmol) was dissolved in dichloromethane (2 mL), hydrochloric acid 1,4-dioxane solution (4 M, 1.88 mL) was added dropwise, and stirred at room temperature overnight. LCMS monitored the reaction was completed, the reaction was concentrated to give white solid 2c (300 mg, yield 74%). ESI-MS (m / z): 233.5 [M+H] + .

[0177] Third step: Compound 1i (50 mg, 197 umol), compound 2c (68 mg, 256 umol) and p-toluenesulfonic acid monohydrate (3.7 mg, 19 umol) were dissolved in n-butanol (2 mL) and reacted at 160 °C in a microwave for 2 hours. LCMS monitored the reaction was completed. The reaction was purified by reverse phase preparative HPLC to give white solid 2 (10.1 mg, yield 11%). ESI-MS (m / z): 449.2 [M+H] + ; 1H NMR (500 MHz, DMSO-d6) δ 8.38 (d, J = 2.8 Hz, 1H), 7.83 (d, J = 8.7 Hz, 1H), 7.46 (dd, J = 8.7, 2.9 Hz, 1H), 6.91 (d, J = 6.9 Hz, 1H), 5.05-5.00 (m, 1H), 4.46-4.42 (m, 1H), 4.12 (q, J = 6.9 Hz, 1H), 4.03-3.99 (m, 2H), 3.28-3.25 (m, 1H), 2.95 (s, 3H), 2.50-2.38 (m, 4H), 1.97-1.87 (m, 1H), 1.84-1.77 (m, 1H), 1.23 (d, J = 6.7 Hz, 3H).

[0178] Example 3

[0179] (R)-4,6-dimethyl-N-(trans-3-(3,4,5-trifluorophenoxy)cyclobutyl)-5,6-dihydro-4H- pyrrolo[3,2,1-de]pyrrolo[2,3-c]carbazin-2-amine

[0180]

[0181] Compound 3 was prepared from the following steps:

[0182]

[0183] First step: Compound 3a (500 mg, 2.66 mmol), compound 3b (946 mg, 3.99 mmol), potassium carbonate (1.47 g, 10.64 mmol) and 18-crown-6 (351 mg, 1.33 mmol) were dissolved in 1,4-dioxane (10 mL) and stirred at 80 °C overnight. The reaction was monitored by LCMS and the reaction was completed. The reaction was diluted with ethyl acetate and washed with water and saturated brine successively. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give yellow solid 3c (600 mg, yield 65%). ESI-MS (m / z): 345.3 [M+H] + .

[0184] Second step: Compound 3c (600 mg, 1.74 mmol) was dissolved in dichloromethane (2 mL) and hydrochloric acid 1,4-dioxane solution (4 M, 2.17 mL) was added dropwise. The reaction was stirred at room temperature overnight. The reaction was monitored by LCMS and the reaction was completed. The reaction was concentrated to give white solid 3d (400 mg, yield 81%). ESI-MS (m / z): 245.3 [M+H] + .

[0185] Step 3: Compound 3d (400 mg, 1.63 mmol) and N, N-diisopropylethylamine (632 mg, 4.90 mmol) were dissolved in 1, 4-dioxane (5 mL) and stirred at 100 °C overnight. LCMS monitored the reaction was completed, the reaction was concentrated, the residue was purified by silica gel column chromatography (dichloromethane / methanol = 10 / 1) to give yellow solid 3e (150 mg, yield 44%). ESI-MS (m / z): 209.4 [M+H] + .

[0186] Step 4: Compound 3e (150 mg, 718 umol), iodomethane (153 mg, 1.08 mmol) and cesium carbonate (468 mg, 1.44 mmol) were dissolved in N, N-dimethylformamide (2 mL) and stirred at 80 °C overnight. LCMS monitored the reaction was completed, the reaction was diluted with ethyl acetate, washed with water and saturated brine successively, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, the residue was purified by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to give yellow solid 3f (100 mg, yield 62%). ESI-MS (m / z): 223.4 [M+H] + .

[0187] Step 5: Compound 3f (30 mg, 134 umol), compound 1e (68 mg, 256 umol) and trifluoroacetic acid (1.5 mg, 13 umol) were dissolved in n-butanol (2 mL) and reacted at 160 °C in microwave for 2 hours. LCMS monitored the reaction was completed. The reaction was purified by reverse phase preparative HPLC to give white solid 3 (6.8 mg, yield 12%). ESI-MS (m / z): 404.2 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) d 8.19 (s, 1H), 7.30 (d, J = 2.8 Hz, 1H), 6.92 - 6.82 (m, 2H), 6.04 (d, J = 2.8 Hz, 1H), 4.93 - 4.81 (m, 1H), 4.46 - 4.41 (m, 1H), 4.37 - 4.24 (m, 1H), 3.64 (dd, J = 12.4, 3.9 Hz, 1H), 3.35 - 3.30 (m, 1H), 3.05 (s, 3H), 2.48 - 2.42 (m, 2H), 2.38 - 2.33 (m, 2H), 1.42 (d, J = 6.4 Hz, 3H).

[0188] Example 4

[0189] (R)-4,6-dimethyl-N-(trans-3-((6-(trifluoromethyl)pyridin-3-yl)oxy)cyclobutyl)-5,6- dihydro-4H-pyrrolo[3,2,1-de]pyrrolo[2,3-b]pyridin-2-amine

[0190]

[0191] Compound 4 was prepared from the following steps:

[0192]

[0193] First step: Compound 3f (30 mg, 134 umol), compound 2b (40 mg, 175 umol) and trifluoroacetic acid (1.5 mg, 13 umol) were dissolved in n-butanol (2 mL) and reacted at 160 °C under microwave for 2 hours. The reaction was monitored by LCMS. The reaction solution was purified by reverse phase preparative HPLC to give white solid 4 (9.3 mg, yield 16%). ESI-MS (m / z): 419.5 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 8.39 (d, J = 2.8 Hz, 1H), 8.20 (s, 1H), 7.84 (d, J = 8.7 Hz, 1H), 7.47 (dd, J = 8.7, 2.9 Hz, 1H), 7.30 (d, J = 2.8 Hz, 1H), 6.69 (br s, 1H), 6.04 (d, J = 2.8 Hz, 1H), 5.08 - 5.02 (m, 1H), 4.53 - 4.45 (m, 1H), 4.35 - 4.25 (m, 1H), 3.64 (dd, J = 12.3, 3.9 Hz, 2H), 3.05 (s, 3H), 2.50 - 2.40 (m, 4H), 1.42 (d, J = 6.4 Hz, 3H).

[0194] Example 5

[0195] (S)-4,6-dimethyl-N-(trans-3-((6-(trifluoromethyl)pyridin-3-yl)oxy)cyclobutyl)-5,6- dihydro-4H-pyrrolo[3,2,1-de]pyrrolo[2,3-b]pyridin-2-amine

[0196]

[0197] Compound 5 was prepared from the following steps:

[0198]

[0199] First step: Compound 3a (792 mg, 4.21 mmol), compound 5a (1 g, 4.21 mmol), potassium carbonate (2.33 g, 16.86 mmol) and 18-crown-6 (557 mg, 2.11 mmol) were dissolved in 1,4-dioxane (10 mL) and stirred at 80 °C overnight. LCMS monitored the reaction was completed, the reaction was diluted with ethyl acetate, washed with water and saturated brine successively, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give yellow solid 5b (1 g, yield 68%). ESI-MS (m / z): 345.3 [M+H] + .

[0200] Second step: Compound 5b (1.0 g, 2.90 mmol) was dissolved in dichloromethane (10 mL), hydrochloric acid 1,4-dioxane solution (4 M, 3.62 mL) was added dropwise, and stirred at room temperature overnight. LCMS monitored the reaction was completed, the reaction was concentrated to give white solid 5c (700 mg, yield 98%). ESI-MS (m / z): 245.4 [M+H] + .

[0201] Third step: Compound 5c (700 mg, 2.86 mmol) and N,N-diisopropylethylamine (1.11 g, 8.57 mmol, 1.49 mL) were dissolved in 1,4-dioxane (10 mL) and stirred at 100 °C overnight. LCMS monitored the reaction was completed, the reaction was concentrated, the residue was purified by silica gel column chromatography (dichloromethane / methanol = 10 / 1) to give yellow solid 5d (250 mg, yield 41%). ESI-MS (m / z): 209.4 [M+H] + .

[0202] Fourth step: Compound 5d (250 mg, 1.20 mmol), iodomethane (153 mg, 1.08 mmol) and cesium carbonate (468 mg, 1.44 mmol) were dissolved in N,N-dimethylformamide (2 mL) and stirred at 80 °C overnight. LCMS monitored the reaction was completed, the reaction was diluted with ethyl acetate, washed with water and saturated brine successively, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, the residue was purified by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to give yellow solid 5e (150 mg, yield 56%). ESI-MS (m / z): 223.4 [M+H] + .

[0203] Fifth step: Compound 5e (30 mg, 134 umol), compound 2b (47 mg, 175 umol) and trifluoroacetic acid (1.5 mg, 13 umol) were dissolved in n-butanol (2 mL) and reacted at 160 °C for 2 hours under microwave. The reaction was monitored by LCMS. The reaction solution was purified by reverse phase preparative HPLC to give white solid 5 (9.2 mg, yield 16%). ESI-MS (m / z): 419.4 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 8.39 (d, J = 2.8 Hz, 1H), 8.22 (s, 1H), 7.84 (d, J = 8.7 Hz, 1H), 7.47 (dd, J = 8.7, 2.8 Hz, 1H), 7.30 (d, J = 2.8 Hz, 1H), 6.74 (br s, 1H), 6.04 (d, J = 2.8 Hz, 1H), 5.10 - 4.99 (m, 1H), 4.51 - 4.45 (m, 1H), 4.31 - 4.25 (m, 1H), 3.64 (d, J = 8.5 Hz, 1H), 3.34 - 3.27 (m, 1H), 3.05 (s, 3H), 2.49 - 2.39 (m, 4H), 1.42 (d, J = 6.4 Hz, 3H).

[0204] Example 6

[0205] (S)-4,6-dimethyl-N-(trans-3-(3,4,5-trifluorophenoxy)cyclobutyl)-5,6-dihydro-4H- pyrrolo[3,2,1-de]pteridin-2-amine

[0206]

[0207] Compound 6 was prepared from the following steps:

[0208]

[0209] First step: Compound 5e (30 mg, 134 umol), compound 1e (44 mg, 175 umol) and trifluoroacetic acid (1.5 mg, 13 umol) were dissolved in n-butanol (2 mL) and reacted at 160 °C for 2 hours under microwave. The reaction was monitored by LCMS. The reaction solution was purified by reverse phase preparative HPLC to give white solid 6 (17.84 mg, yield 32%). ESI-MS (m / z): 404.2 [M+H] + ; 1H NMR (400 MHz, DMSO-d6) δ 8.22 (s, 1H), 7.31 (d, J = 2.8 Hz, 1H), 6.91 - 6.84 (m, 2H), 6.05 (d, J = 2.8 Hz, 1H), 4.94 - 4.80 (m, 1H), 4.45 - 4.40 (m, 1H), 4.31 - 4.25 (m, 1H), 3.68 - 3.63 (m, 1H), 3.31 (dd, J = 12.4, 8.2 Hz, 1H), 3.05 (s, 3H), 2.47 - 2.29 (m, 4H), 1.42 (d, J = 6.4 Hz, 3H).

[0210] Example 7

[0211] (S)-4,5-Dimethyl-2-((cis-3-((6-(trifluoromethyl)pyridin-3-yl)amino)cyclobutyl)amino)- 4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0212]

[0213] Compound 7 was prepared from the following steps:

[0214]

[0215] First step: 5-bromo-2-trifluoromethylpyridine 7a (452 mg, 2.00 mmol), cis-3-amino-1- cyclobutylcarbamic acid tert-butyl ester 7b (372 mg, 2.00 mmol), Pd2(dba)3 (91.5 mg, 0.10 mmol), Xantphos (115.6 mg, 0.20 mmol) and cesium carbonate (1.3 g, 4.00 mmol) were dissolved in dioxane (10 mL) and stirred at 100 °C overnight under nitrogen protection. TLC monitored the reaction was completed, the reaction was cooled to room temperature, filtered, the filter cake was washed with dichloromethane twice, the filtrate was concentrated, the residue was purified by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to give white solid 7c (580 mg, yield 87%). ESI-MS (m / z): 332.3 [M+H] + .

[0216] Second step: Compound 7c (100 mg, 0.3 mmol) was dissolved in methanol (5 mL), hydrogen chloride-dioxane solution (4 M, 0.75 mL, 3 mmol) was added dropwise at room temperature, stirred at room temperature for 2 hours. TLC monitored the reaction was completed, the reaction was rotary evaporated to give white solid 7d (80 mg, yield 99%). ESI-MS (m / z): 232.4 [M+H] + .

[0217] Step 3: Compound 1i (76 mg, 0.30 umol), compound 7d (80 mg, 0.30 mol) and p-toluenesulfonic acid monohydrate (5.7 mg, 0.03 mol) were dissolved in n-butanol (3 mL) and reacted at 160 °C for 2 hours in a microwave. The reaction was monitored by LCMS. The reaction solution was purified by reverse phase preparative HPLC to give white solid 7 (45 mg, yield 33%). ESI-MS (m / z): 448.5 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) d 8.01 (d, J = 2.7 Hz, 1H), 7.51 (d, J = 8.7 Hz, 1H), 6.98 - 6.90 (m, 2H), 6.81 (d, J = 7.5 Hz, 1H), 4.19 - 3.97 (m, 3H), 3.58 (q, J = 7.4 Hz, 1H), 3.32 - 3.21 (m, 1H), 2.96 (s, 3H), 2.83 - 2.70 (m, 2H), 2.55 - 2.52 (m, 2H), 1.98 - 1.88 (m, 1H), 1.86 - 1.73 (m, 3H), 1.23 (d, J = 6.8 Hz, 3H).

[0218] Example 8

[0219] (S)-4,5-Dimethyl-2-((trans-3-((6-(trifluoromethyl)pyridin-3-yl)amino)cyclobutyl)amino)- 4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0220]

[0221] Compound 8 was obtained by replacing 7b in the first step of Example 7 with (trans-3-aminocyclobutyl)carbamic acid tert-butyl ester, using similar methods and reaction procedures. ESI-MS (m / z): 448.5 [M+H] + ; 1H NMR (400 MHz, DMSO-d6) δ 7.98 (d, J = 2.7 Hz, 1H), 7.53 (d, J = 8.6 Hz, 1H), 7.08 (d, J = 5.5 Hz, 1H), 6.92 - 6.80 (m, 2H), 4.44 (q, J = 7.3 Hz, 1H), 4.12 (q, J = 6.7 Hz, 1H), 4.07 - 3.98 (m, 1H), 3.96 - 3.86 (m, 1H), 3.32 - 3.22 (m, 1H), 2.95 (s, 3H), 2.42 - 2.28 (m, 2H), 2.25 - 2.14 (m, 2H), 1.97 - 1.87 (m, 1H), 1.85 - 1.72 (m, 1H), 1.23 (d, J = 6.8 Hz, 3H).

[0222] Example 9

[0223] (S)-4,5-Dimethyl-2-((trans-3-((6-(trifluoromethyl)pyridin-3-yl)amino)cyclopentyl)amino)- 4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0224]

[0225] Example 9 using similar methods and reaction procedures. ESI-MS (m / z): 462.4 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 8.04 (d, J = 2.7 Hz, 1H), 7.51 (d, J = 8.7 Hz, 1H), 6.98 (dd, J = 8.7, 2.8 Hz, 1H), 6.74 (d, J = 6.6 Hz, 1H), 6.56 (d, J = 7.6 Hz, 1H), 4.29 (q, J = 6.9 Hz, 1H), 4.11 (q, J = 6.7 Hz, 1H), 4.03 (dt, J = 13.0, 4.7 Hz, 1H), 3.92 (q, J = 6.4 Hz, 1H), 3.30 - 3.22 (m, 1H), 2.93 (s, 3H), 2.53 (d, J = 5.0 Hz, 2H), 2.19 - 2.02 (m, 2H), 1.91 (p, J = 6.9 Hz, 2H), 1.80 (dt, J = 12.7, 6.7 Hz, 2H), 1.58 - 1.41 (m, 2H), 1.23 (d, J = 6.8 Hz, 3H).

[0226] Example 10

[0227] (S)-4,5-dimethyl-2-((1-((6-(trifluoromethyl)pyridin-3-yl)methyl)azetidin-3-yl)amino)- 4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0228]

[0229] Compound 10 was prepared by the following steps:

[0230]

[0231] First Step: Compound 1i (200 mg, 791 umol) and compound 10a (451 mg, 1.58 mmol) were dissolved in 10 mL of dioxane, Pd2dba3 (72 mg, 79 umol), S-Phos (65 mg, 158 umol) and sodium tert-butoxide (228 mg, 2.37 mmol) were added, and the reaction was stirred at 100 °C overnight under nitrogen protection. The reaction was monitored by LCMS until the starting material was consumed. The reaction was concentrated, and the residue was purified by preparative thin layer chromatography (dichloromethane / methanol = 20 / 1) to give compound 10b (90 mg, 29% yield) as a yellow oil. ESI-MS (m / z): 389.5 [M+H] + .

[0232] Second Step: Compound 10b (90 mg, 232 umol) was dissolved in 10 mL of dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction was stirred at room temperature for 4 hours. The reaction was monitored by LCMS until the starting material was consumed. The reaction was concentrated to give compound 10c (90 mg) as a yellow oil, which was used directly in the next step. ESI-MS (m / z): 289.4 [M+H] + .

[0233] Third Step: Compound 10c (90 mg) and 10d (53 mg, 269 umol) were dissolved in 10 mL of acetonitrile, and potassium carbonate (93 mg, 673 umol) was added. The reaction was stirred at 60 °C for 4 hours. The reaction was monitored by LCMS until the starting material was consumed. The reaction was concentrated, and the residue was purified by reverse phase preparative HPLC to give compound 10 (10 mg, 10.3% yield) as a white solid. ESI-MS (m / z): 448.5 [M+H] + ; 1H NMR (500 MHz, DMSO-d6) δ 8.66 (s, 1H), 7.96 (d, J = 8.0 Hz, 1H), 7.85 (d, J = 8.0 Hz, 1H), 6.89 (d, J = 6.9 Hz, 1H), 4.40 (q, J = 7.0 Hz, 1H), 4.12 (q, J = 6.8 Hz, 1H), 4.01 (dt, J = 13.0, 4.9 Hz, 1H), 3.72 (s, 2H), 3.59 (t, J = 6.8 Hz, 2H), 3.30 - 3.23 (m, 1H), 2.95 (d, J = 8.0 Hz, 5H), 2.52 (s, 2H), 1.92 (d, J = 14.5 Hz, 1H), 1.80 (d, J = 3.9 Hz, 1H), 1.23 (d, J = 6.7 Hz, 3H).

[0234] Example 11

[0235] (S)-4,5-Dimethyl-2-(((1s,3R)-3-(((6-(trifluoromethyl)pyridin-3-yl)oxy)methyl)cyclobutyl)amino)- 4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0236]

[0237] Compound 11 was prepared from the following steps:

[0238]

[0239] First step: cis-tert-butyl 3-hydroxymethylcyclobutylcarbamate 11a (150 mg, 0.745 mmol), methanesulfonic anhydride (259 mg, 1.49 mmol) and N,N-diisopropylethylamine (385 mg, 2.98 mmol) were dissolved in dichloromethane (5 mL) and stirred at room temperature overnight. The reaction was monitored by TLC and upon completion, the reaction was diluted with dichloromethane and washed with water and saturated brine successively. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give yellow solid 11b (204 mg, yield 98%). 1 H NMR (500 MHz, DMSO-d6) δ 7.10 (d, J = 8.1 Hz, 1H), 4.11 (d, J = 5.2 Hz, 2H), 3.19 (d, J = 1.6 Hz, 1H), 3.16 (d, J = 1.6 Hz, 3H), 2.26 (d, J = 5.9 Hz, 1H), 1.68 (d, J = 8.7 Hz, 2H), 1.57 - 1.47 (m, 2H), 1.37 (s, 9H).

[0240] Second step: Compound 11b (204 mg, 731 umol), compound 2a (131 mg, 0.8 mmol) and cesium carbonate (485 mg, 1.49 mmol) were dissolved in N,N-dimethylformamide (5 mL) and stirred at 90 °C overnight. TLC monitored the end of the reaction, the reaction liquid was diluted with water, extracted with ethyl acetate, washed with saturated brine, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1) to obtain yellow solid 11c (185 mg, yield 72%). ESI-MS (m / z): 347.2 [M+H] + .

[0241] Third step: Compound 11c (120 mg, 346 umol) was dissolved in dichloromethane (5 mL), 4M hydrochloric acid dioxane solution (0.87 mL) was added at 0 °C and stirred overnight. TLC monitored the end of the reaction, directly rotary dried to obtain white solid 11d (85 mg, yield 99%). ESI-MS (m / z): 247.4 [M+H] + .

[0242] Fourth step: Compound 11d (82 mg, 336 umol), compound 1i (85 mg, 336 umol) and p-toluenesulfonic acid monohydrate (5.7 mg, 33.6 umol) were dissolved in n-butanol (3 mL) and reacted at 160 °C for 3 hours under microwave. LCMS monitored the end of the reaction. The reaction liquid was purified by reverse phase preparative HPLC to obtain white solid 11 (25 mg, yield 16%). ESI-MS (m / z): 463.3 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) d 8.47 (d, J = 3.0 Hz, 1H), 7.85 (d, J = 8.5 Hz, 1H), 7.64 (dd, J = 9.0, 3.0 Hz, 1H), 6.81 (br s, 1H), 4.32-4.23 (m, 1H), 4.16-4.09 (m, 3H), 4.08-4.01 (m, 1H), 3.30-3.24 (m, 2H), 2.95 (s, 3H), 2.48-2.38 (m, 3H), 1.97-1.88 (m, 1H), 1.86-1.73 (m, 3H), 1.23 (d, J = 7.0 Hz, 3H).

[0243] Example 12

[0244] (S)-2-(((1s,3R)-3-(((1-cyclopropyl-3-(trifluoromethyl)-1H-pyrazol-5-yl)oxy)methyl)cyclobutyl)amino)-4,5-dimethyl-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0245]

[0246] Compound 12 was prepared by the following steps:

[0247]

[0248] First step: ethyl trifluoroacetoacetate 12a (1.7 g, 9.21 mmol) and cyclopropylhydrazine hydrochloride 12b (1.0 g, 9.21 mmol) were dissolved in 20 mL of ethanol and reacted at 80 °C overnight. The reaction solution was concentrated, the residue was slurried with petroleum ether, and compound 12c (800 mg, yield 45%) was obtained as a brown solid by filtration. ESI-MS (m / z): 193.2 [M+H] + .

[0249] Second step: compound 11b (872 mg, 3.12 mmol), compound 12c (500 mg, 2.61 mmol) and cesium carbonate (1.70 g, 5.22 mmol) were dissolved in N,N-dimethylformamide (20 mL) and stirred at 90 °C overnight. The reaction was monitored by TLC and the reaction solution was diluted with water, extracted with ethyl acetate, washed with saturated brine, and the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1) to obtain compound 12d (495 mg, yield 52%) as a yellow solid. ESI-MS (m / z): 362.6 [M+H] + .

[0250] Third step: compound 12d (495 mg, 1.1 mmol) was dissolved in dichloromethane (30 mL), 4M hydrochloric acid dioxane solution (1.37 mL) was added at 0 °C and stirred overnight. The reaction was monitored by TLC and directly rotary evaporated to obtain compound 12e (231 mg, yield 80%) as a white solid. ESI-MS (m / z): 262.6 [M+H] + .

[0251] Fourth step: Compound 12e (26 mg, 95 umol), compound 1i (20 mg, 79 umol) and p-toluenesulfonic acid monohydrate (0.4 mg, 7.9 umol) were dissolved in n-butanol (3 mL) and reacted at 160 °C for 3 hours under microwave. The reaction was monitored by LCMS. The reaction solution was purified by reverse phase preparative HPLC to obtain white solid 12 (13 mg, yield 35%). ESI-MS (m / z): 492.4 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) δ 6.67 (br s, 1H), 6.17 (s, 1H), 4.26-4.18 (m, 1H), 4.15-4.06 (m, 2H), 4.05-3.97 (m, 1H), 3.54-3.21 (m, 5H), 2.94 (s, 3H), 2.60-2.50 (m, 2H), 2.47-2.35 (m, 3H), 1.96-1.86 (m, 1H), 1.85-1.70 (m, 3H), 1.21 (d, J = 6.8 Hz, 3H), 1.05-0.90 (m, 4H).

[0252] Example 13

[0253] (S)-2-(((1s,3R)-3-(((1-cyclopropyl-3-(trifluoromethyl)-1H-pyrazol-5-yl)oxy)methyl)cyclobutyl)amino)-5-(hydroxymethyl)-4,5-dimethyl-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1- de]pteridin-6-one

[0254]

[0255] Compound 13 was prepared from the following steps:

[0256]

[0257] First step: 2,4-dichloropyrido[3,2-d]pyrimidine 1f (4.0 g, 20.0 mmol) and 2-methyl-L- serine methyl ester hydrochloride 13a (4.07 g, 24.0 mmol) were dissolved in dichloromethane (30 mL), N,N-diisopropylethylamine (7.75 g, 59.99 mmol, 10.45 mL) was added, and stirred at room temperature overnight. The reaction was monitored by LCMS, the reaction solution was diluted with dichloromethane, washed with water and saturated brine respectively, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain white solid 13b (5.0 g, yield 84%). ESI-MS (m / z): 297.3 [M+H] + .

[0258] Second Step: Compound 13b (5.0 g, 16.85 mmol) was dissolved in tetrahydrofuran (50 mL), hydrochloric acid aqueous solution (6 M, 5.62 mL) and platinum dioxide (382 mg, 1.69 mmol) were added, the reaction system was replaced with hydrogen balloon, stirred at room temperature under the pressure of hydrogen balloon for 48 hours, LCMS monitoring showed that the reaction was completed. The reaction solution was diluted with methanol, filtered, and the filtrate was concentrated to obtain white solid 13c (4.0 g, yield 88%). ESI-MS (m / z): 269.3 [M+H] + .

[0259] Third Step: Compound 13c (1.5 g, 5.58 mmol) and methyl iodide (1.58 g, 11.16 mmol) were dissolved in acetonitrile (5 mL), cesium carbonate (3.64 g, 11.16 mmol) was added, and the reaction mixture was stirred at room temperature for 48 hours. LCMS monitoring showed that the reaction was completed. The reaction solution was diluted with ethyl acetate, filtered, washed with water and saturated brine, and the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (dichloromethane / methanol = 10 / 1) to obtain yellow solid 13d (1.1 g, yield 69%). ESI-MS (m / z): 283.3 [M+H] + .

[0260] Fourth Step: Compound 12e (25 mg, 84 umol), compound 13d (20 mg, 70.7 umol) and p-toluenesulfonic acid monohydrate (0.4 mg, 7.07 umol) were dissolved in n-butanol (3 mL), and the reaction was carried out at 160°C for 3 hours under microwave. LCMS monitoring showed that the reaction was completed. The reaction solution was purified by reverse phase preparative HPLC to obtain white solid 13 (13 mg, yield 35%). ESI-MS (m / z): 522.3 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) δ 6.54 (br s, 1H), 6.17 (s, 1H), 5.06 (t, J = 5.5 Hz, 1H), 4.29-4.18 (m, 1H), 4.10 (d, J = 4.9 Hz, 2H), 3.76-3.67 (m, 2H), 3.65-3.58 (m, 1H), 3.57-3.49 (m, 2H), 2.97 (s, 3H), 2.55-2.45 (m, 2H), 2.44-2.32 (m, 3H), 1.88-1.72 (m, 4H), 1.33 (s, 3H), 0.98 (d, J = 7.8 Hz, 4H).

[0261] Example 14

[0262] (S)-5-(hydroxymethyl)-4,5-dimethyl-2-(((1s,3R)-3-(((2-methyl-6- (trifluoromethyl)pyridin-3-yl)oxy)methyl)cyclobutyl)amino)-4,5,9,10-tetrahydro- 6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0263]

[0264] Compound 14 was prepared from the following steps:

[0265]

[0266] First step: Compound 2a (2.0 g, 12.26 mmol), Na2CO3(2.6 g, 24.52 mmol) were added to water (60 mL) and stirred to dissolve. Iodine (3.11 g, 12.26 mmol) was added to the reaction solution, which was stirred at room temperature for 3 hours. LCMS was used to monitor the disappearance of the starting material. The reaction solution was adjusted to pH 5-6 with dilute hydrochloric acid, and extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was subjected to silica gel column chromatography (ethyl acetate / petroleum ether = 0-30% gradient elution) to obtain white solid 14a (1.5 g, yield 42%). ESI-MS (m / z): 290.2 [M+H] + .

[0267] Second step: Compound 14a (0.5 g, 1.73 mmol), benzyl bromide (337 mg, 1.98 mmol) were dissolved in DMF (5 mL), and K2CO3(364 mg, 2.64 mmol) was added. The reaction solution was stirred at 50°C for 2 hours, and LCMS was used to monitor the end of translation. The reaction solution was diluted with water, and extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was subjected to silica gel column chromatography (ethyl acetate / petroleum ether = 0-15% gradient elution) to obtain white solid 14b (0.5 g, yield 76%). ESI-MS (m / z): 380.2 [M+H] + .

[0268] Step 3: Compound 14b (412 mg, 1.09 mmol), palladium acetate (24 mg, 109 umol), trimethylboroxine (682 mg, 5.43 mmol), tricyclohexylphosphine (152 mg, 523 umol), potassium phosphate tribasic (922 mg, 4.35 mmol), water (3 mL), 1,4-dioxane (30 mL) were added into a 100 mL two-necked flask, after replaced with nitrogen, stirred at 90 °C overnight, TLC monitored the reaction was completed, the reaction solution was diluted with water, filtered with celite, extracted with ethyl acetate, finally washed with saturated brine, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1) to give yellow liquid 14c (198 mg, yield 68%). ESI-MS (m / z): 268.2 [M+H]+.

[0269] Step 4: Compound 14c (198 mg, 741 umol) was dissolved in dichloromethane (10 mL), boron tribromide (928 mg, 3.71 umol) was added dropwise at -78 °C and stirred for 1 hour, TLC monitored the reaction was completed, the reaction solution was diluted with water, extracted with dichloromethane, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give yellow liquid 14d (143 mg, yield 91%). ESI-MS (m / z): 178.4 [M+H]+.

[0270] Step 5: Compound 14d (120 mg, 677 umol), compound 11b (208 mg, 745 umol) and cesium carbonate (441 mg, 1.35 mmol) were dissolved in N,N-dimethylformamide (5 mL), stirred at 90 °C overnight. TLC monitored the reaction was completed, the reaction solution was diluted with water, extracted with ethyl acetate, washed with saturated brine, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1) to give yellow solid 14e (196 mg, yield 80%). ESI-MS (m / z): 361.4 [M+H] + .

[0271] Step 6: Compound 14e (196 mg, 543 umol) was dissolved in dichloromethane (10 mL), 4M hydrochloric acid (0.952 mL) was added at 0 °C and stirred overnight. TLC monitored the reaction was completed, directly rotary evaporation to give white solid 14f (112 mg, yield 79%). ESI-MS (m / z): 261.3 [M+H] + .

[0272] Seventh step: Compound 14f (27 mg, 106 umol), compound 13d (20 mg, 70.7 umol) and p-toluenesulfonic acid monohydrate (0.4 mg, 7.07 umol) were dissolved in n-butanol (3 mL) and reacted at 160 °C for 3 hours under microwave. The reaction was monitored by LCMS. The reaction solution was purified by reverse phase preparative HPLC to give white solid 14 (19 mg, yield 52%). ESI-MS (m / z): 507.3 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) d 7.67 (d, J = 8.5 Hz, 1H), 7.48 (d, J = 8.5 Hz, 1H), 6.54 (br s, 1H), 5.06 (t, J = 5.5 Hz, 1H), 4.29 - 4.19 (m, 1H), 4.05 (d, J = 4.8 Hz, 2H), 3.77 - 3.66 (m, 2H), 3.65 - 3.60 (m, 1H), 3.58 - 3.53 (m, 1H), 2.98 (s, 3H), 2.48 - 2.35 (m, 8H), 1.90 - 1.74 (m, 4H), 1.33 (s, 3H).

[0273] Example 15

[0274] (S)-2-(((1s,3R)-3-(((2-cyclopropyl-6-(trifluoromethyl)pyridin-3-yl)oxy)methyl)cyclobutyl)amino)-5-(hydroxymethyl)-4,5-dimethyl-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0275]

[0276] Compound 15 was prepared from the following steps:

[0277]

[0278] First step: Compound 14b (300 mg, 0.791 mmol), palladium acetate (18 mg, 79 umol), cyclopropylboronic acid (339 mg, 3.96 mmol), tricyclohexylphosphine (152 mg, 523 umol), potassium phosphate tribasic (672 mg, 3.17 mmol), water (1 mL), 1,4-dioxane (10 mL) were added into a 100 mL two-necked flask, after replaced with nitrogen, stirred at 90 °C overnight, TLC monitored the reaction was completed, the reaction solution was diluted with water, filtered with diatomite, extracted with ethyl acetate, finally washed with saturated brine, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1) to give yellow liquid 15a (209 mg, yield 90%). ESI-MS (m / z): 294.3 [M+H]+.

[0279] Second step: Compound 15a (209 mg, 712 umol) was dissolved in dichloromethane (10 mL), and boron tribromide (829 mg, 3.56 umol) was added dropwise at -78 °C and stirred for 1 hour, TLC monitored the reaction was completed, the reaction solution was diluted with water, extracted with dichloromethane, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give yellow liquid 15b (122 mg, yield 84%). ESI-MS (m / z): 204.4 [M+H]+.

[0280] Third step: Compound 15b (122 mg, 590 umol), compound 11b (211 mg, 758 umol) and cesium carbonate (449 mg, 1.38 mmol) were dissolved in N,N-dimethylformamide (5 mL), stirred at 90 °C overnight. TLC monitored the reaction was completed, the reaction solution was diluted with water, extracted with ethyl acetate, washed with saturated brine, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1) to give yellow solid 15c (152 mg, yield 66%). ESI-MS (m / z): 387.3 [M+H]+ + .

[0281] Fourth step: Compound 15c (152 mg, 393 umol) was dissolved in dichloromethane (10 mL), 4M hydrochloric acid (0.706 mL) was added at 0 °C and stirred overnight. TLC monitored the reaction was completed, directly rotary evaporation to give white solid 15d (92 mg, yield 79%). ESI-MS (m / z): 287.3 [M+H] + .

[0282] Step 5: Compound 15d (46 mg, 159 umol), compound 13d (30 mg, 106 umol) and p-toluenesulfonic acid monohydrate (0.5 mg, 10.6 umol) were dissolved in n-butanol (3 mL) and reacted at 160 °C for 3 hours under microwave. The reaction was monitored by LCMS. The reaction solution was purified by reverse phase preparative HPLC to give white solid 15 (18 mg, yield 32%). ESI-MS (m / z): 533.1 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) d 7.56 (dd, J = 8.5, 3.8 Hz, 1H), 7.45 (d, J = 8.5 Hz, 1H), 6.55 (br s, 1H), 5.06 (t, J = 5.7 Hz, 1H), 4.30 - 4.18 (m, 1H), 4.06 (d, J = 5.1 Hz, 2H), 3.77 - 3.66 (m, 2H), 3.65 - 3.59 (m, 1H), 3.57 - 3.52 (m, 1H), 2.98 (s, 3H), 2.52 - 2.45 (m, 2H), 2.44 - 2.35 (m, 3H), 1.87 - 1.75 (m, 4H), 1.33 (s, 3H), 1.02 - 0.97 (m, 2H), 0.95 - 0.89 (m, 2H).

[0283] Example 16

[0284] (S)-5-(Methoxymethyl)-4-(methyl-d3)-2-(((1s,3R)-3-(((6-(trifluoromethyl)pyridin-3- yl)oxy)methyl)cyclobutyl)amino)-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6- one

[0285]

[0286] Compound 16 was prepared from the following steps:

[0287]

[0288] First Step: Dissolve 2,4-dichloropyrido[3,2-d]pyrimidine 1f (2 g, 10.00 mmol) and O-methyl-L-serine methyl ester hydrochloride 16a (2.45 g, 15.00 mmol) in tetrahydrofuran (20 mL), add N,N-diisopropylethylamine (3.88 g, 30.00 mmol, 5.22 mL), stir at room temperature overnight. Monitor the reaction by LCMS until completion. Concentrate the reaction mixture and purify the residue by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give yellow oil 16b (2.5 g, 84% yield). ESI-MS (m / z): 297.2 [M+H] + .

[0289] Second Step: Dissolve compound 16b (2.5 g, 8.43 mmol) in tetrahydrofuran (30 mL), add hydrochloric acid aqueous solution (6 N, 1.40 mL) and platinum dioxide (191 mg, 0.84 mmol), replace the reaction system with a hydrogen balloon with hydrogen, stir at room temperature under the pressure of the hydrogen balloon for 48 hours. Monitor the reaction by LCMS until completion. Dilute the reaction mixture with methanol, filter with celite, concentrate the filtrate and purify by silica gel column chromatography (dichloromethane / methanol = 10 / 1) to give gray solid 16c (2.0 g, 88% yield). ESI-MS (m / z): 269.3 [M+H] + .

[0290] Third Step: Dissolve compound 16c (200 mg, 0.74 mmol) and deuterated methyl iodide (215 mg, 1.49 mmol) in acetonitrile (5 mL), add cesium carbonate (485 mg, 1.49 mmol), react at 50 °C for 2 hours. Monitor the reaction by LCMS until completion. Dilute the reaction mixture with dichloromethane, filter, concentrate the filtrate and purify by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to give yellow solid 16d (200 mg, 94% yield). ESI-MS (m / z): 286.3 [M+H] + .

[0291] Fourth Step: Dissolve compound 16d (25 mg, 89 umol), compound 11c (32 mg, 131 umol) and p-toluenesulfonic acid monohydrate (0.45 mg, 8.9 umol) in n-butanol (3 mL), react at 160 °C under microwave for 3 hours. Purify the reaction mixture by reverse phase preparative HPLC to give white solid 16 (5 mg, 10% yield). ESI-MS (m / z): 496.4 [M+H] + ; 1H NMR (500 MHz, DMSO-d6) δ 8.43 (d, J = 2.7 Hz, 1H), 7.82 (d, J = 8.7 Hz, 1H), 7.59 (dd, J = 8.6, 2.8 Hz, 1H), 6.53 (br s, 1H), 5.06 (t, J = 5.5 Hz, 1H), 4.29 - 4.18 (m, 1H), 4.10 (d, J = 4.9 Hz, 2H), 3.78 - 3.66 (m, 2H), 3.65 - 3.58 (m, 1H), 3.57 - 3.51 (m, 1H), 2.98 (s, 3H), 2.52 - 2.46 (m, 2H), 2.45 - 2.34 (m, 3H), 1.90 - 1.73 (m, 4H), 1.33 (s, 3H).

[0292] Example 17

[0293] (S)-5-(hydroxymethyl)-4,5-dimethyl-2-(((1s,3R)-3-(((6- (trifluoromethyl)pyridin-3-yl)oxy)methyl)cyclobutyl)amino)-4,5,9,10-tetrahydro- 6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0294]

[0295] Replace the starting material 14d in the fifth step of Example 14 with 6- (trifluoromethyl)pyridin-3-ol 2a, using similar methods and reaction procedures to obtain compound 17. ESI-MS (m / z): 493.3 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) δ 8.43 (d, J = 2.7 Hz, 1H), 7.82 (d, J = 8.7 Hz, 1H), 7.59 (dd, J = 8.6, 2.8 Hz, 1H), 6.53 (br s, 1H), 5.06 (t, J = 5.5 Hz, 1H), 4.29 - 4.18 (m, 1H), 4.10 (d, J = 4.9 Hz, 2H), 3.78 - 3.66 (m, 2H), 3.65 - 3.58 (m, 1H), 3.57 - 3.51 (m, 1H), 2.98 (s, 3H), 2.52 - 2.46 (m, 2H), 2.45 - 2.34 (m, 3H), 1.90 - 1.73 (m, 4H), 1.33 (s, 3H).

[0296] Example 18

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

[0298]

[0299] Compound 18 was obtained by replacing 2a in the first step of Example 11 with 2- (trifluoromethyl)pyrimidin-5-ol, using similar method and reaction procedures. ESI-MS (m / z): 464.3 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) δ 8.76 (s, 2H), 6.70 (s, 1H), 4.28-4.17 (m, 3H), 4.11 (q, J = 6.8 Hz, 1H), 4.11-4.39 (m, 1H), 3.33 (s, 1H), 3.30-3.22 (m, 2H), 2.94 (s, 3H), 2.52-2.46 (m, 2H), 2.45-2.35 (m, 3H), 1.95-1.86 (m, 1H), 1.85-1.72 (m, 3H), 1.21 (d, J = 6.7 Hz, 3H).

[0300] Example 19

[0301] (S)-5-(hydroxymethyl)-4,5-dimethyl-2-(((1s,3R)-3-((3,4,5-trifluorophenoxy)methyl)cyclobutyl)amino)-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0302]

[0303] Compound 19 was obtained by replacing starting material 14d in the fifth step of Example 14 with 3,4,5-trifluorophenol 1c, using similar method and reaction procedures. ESI-MS (m / z): 478.4 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) δ 7.10-6.95 (m, 2H), 6.48 (d, J = 7.8 Hz, 1H), 5.06 (br s, 1H), 4.24 (q, J = 8.0 Hz, 1H), 3.94 (d, J = 5.9 Hz, 2H), 3.81-3.70 (m, 2H), 3.69-3.64 (m, 1H), 3.57 (d, J = 11.3 Hz, 1H), 3.00 (s, 3H), 2.52-2.45 (m, 2H), 2.43-2.33 (m, 3H), 1.90-1.80 (m, 2H), 1.79-1.71 (m, 2H), 1.35 (s, 3H).

[0304] Example 20

[0305] (S)-2-(((1s,3R)-3-((4-fluoro-2-(trifluoromethyl)phenoxy)methyl)cyclobutyl)amino)- 5-(hydroxymethyl)-4,5-dimethyl-4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1- rev]pteridin-6-one

[0306]

[0307] Compound 20 was obtained by replacing the starting material 14d in the fifth step of Example 14 with 4-fluoro-2-(trifluoromethyl)phenol, using similar methods and reaction procedures. ESI-MS (m / z): 510.4 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) δ 7.60-7.50 (m, 2H), 7.34-7.27 (m, 1H), 6.43 (br s, 1H), 5.08 (t, J = 5.6 Hz, 1H), 4.22 (q, J = 8.2 Hz, 1H), 4.06 (d, J = 5.2 Hz, 2H), 3.77-3.68 (m, 2H), 3.67-3.62 (m, 1H), 3.61-3.55 (m, 1H), 2.99 (s, 3H), 2.51-2.45 (m, 2H), 2.43-2.35 (m, 3H), 1.90-1.83 (m, 2H), 1.82-1.71 (m, 2H), 1.34 (s, 3H).

[0308] Example 21

[0309] (S)-5-(hydroxymethyl)-4,5-dimethyl-2-(((1s,3R)-3-(((2-phenyl-6- (trifluoromethyl)pyridin-3-yl)oxy)methyl)cyclobutyl)amino)-4,5,9,10-tetrahydro- 6H,8H-pyrido[3,2,1-rev]pteridin-6-one

[0310]

[0311] Compound 21 was prepared by the following steps:

[0312]

[0313] First step: Compound 21a (290 mg, 1.46 mmol), compound 11b (451 mg, 1.61 mmol) and cesium carbonate (191 mg, 2.92 mmol) were dissolved in N,N-dimethylformamide (5 mL) and stirred at 90 °C overnight. TLC monitored the end of the reaction, the reaction liquid was diluted with water, extracted with ethyl acetate, washed with saturated brine, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1) to obtain yellow solid 21b (445 mg, yield 79%). ESI-MS (m / z): 381.3 [M+H] + .

[0314] Second step: Compound 21b (158 mg, 414 umol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (30 mg, 41.4 umol), phenylboronic acid (101 mg, 829 umol), cesium carbonate (405 mg, 1.24 mmol), water (1 mL), dioxane (10 mL) were added to a 25 mL two-necked flask, stirred at 90 °C overnight after nitrogen replacement, TLC monitored the end of the reaction, the reaction liquid was diluted with water, filtered with diatomite, extracted with ethyl acetate, finally washed with saturated brine, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1) to obtain yellow liquid 21c (152 mg, yield 87%). ESI-MS (m / z): 423.4 [M+H] + .

[0315] Third step: Compound 21c (78 mg, 164 umol) was dissolved in dichloromethane (5 mL), 4M hydrochloric acid dioxane solution (0.1 mL) was added at 0 °C and stirred overnight. TLC monitored the end of the reaction, directly rotary evaporation to obtain white solid 21d (43 mg, yield 72%). ESI-MS (m / z): 323.7 [M+H] + .

[0316] Fourth step: Compound 21d (43 mg, 133 umol), compound 13d (25 mg, 88.4 umol) and p-toluenesulfonic acid monohydrate (1.31 mg, 13.3 umol) were dissolved in n-butanol (3 mL) and reacted at 160 °C for 3 hours under microwave. LCMS monitored the end of the reaction. The reaction liquid was purified by reverse phase preparative HPLC to obtain white solid 21 (14 mg, yield 27%). ESI-MS (m / z): 569.9 [M+H] + ; 1H NMR (500 MHz, DMSO-d6) δ 7.92 (d, J = 7.7 Hz, 2H), 7.83 (d, J = 8.6 Hz, 1H), 7.74 (d, J = 8.6 Hz, 1H), 7.60 - 7.41 (m, 2H), 7.44 (d, J = 7.2 Hz, 1H), 6.40 (br s, 1H), 5.06 (t, J = 5.6 Hz, 1H), 4.23 (p, J = 8.2 Hz, 1H), 4.13 (d, J = 5.3 Hz, 2H), 3.79 - 3.68 (m, 2H), 3.66 - 3.61 (m, 1H), 3.58 - 3.52 (m, 1H), 2.98 (s, 3H), 2.52 - 2.35 (m, 5H), 1.90 - 1.72 (m, 4H), 1.33 (s, 2H).

[0317] Example 22

[0318] (S)-5-(hydroxymethyl)-4,5-dimethyl-2-(((1s,3R)-3-(((2-(pyrrolidin-1-yl)-6- (trifluoromethyl)pyridin-3-yl)oxy)methyl)cyclobutyl)amino)-4,5,9,10-tetrahydro- 6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0319]

[0320] Compound 22 was prepared from the following steps:

[0321]

[0322] First Step: Compound 14b (210 mg, 554 umol), Pd2(dba)3 (50.7 mg, 55.4 umol), tetrahydro pyrrole (89 mg, 830 umol), potassium tert-butoxide (310 mg, 2.77 mmol), 2-dicyclohexylphosphino-2'-(N,N-dimethylamine)-biphenyl (65 mg, 166 umol) and toluene (8 mL) were added into a 50 mL single neck flask, the reaction system was replaced by nitrogen and stirred at 80 °C overnight, LCMS showed that the reaction was completely converted. The reaction solution was filtered with diatomite, then extracted with ethyl acetate and water, washed with saturated brine, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1) to obtain yellow liquid 22a (132 mg, yield 73%). ESI-MS (m / z): 323.3 [M+H]+.

[0323] Second step: Compound 22a (132 mg, 409 umol) was dissolved in dichloromethane (10 mL), and boron tribromide (474 mg, 1.89 umol) was added dropwise at -78 °C and stirred for 1 hour. TLC monitoring showed that the reaction was completed. The reaction solution was diluted with water and extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to obtain yellow liquid 22b (65 mg, yield 73%). ESI-MS (m / z): 233.3 [M+H]+.

[0324] Third step: Compound 22b (65 mg, 279 umol) and compound 11b (86 mg, 307 umol) and cesium carbonate (189 mg, 560 umol) were dissolved in N,N-dimethylformamide (5 mL) and stirred at 90 °C overnight. TLC monitoring showed that the reaction was completed. The reaction solution was diluted with water and extracted with ethyl acetate, and then washed with saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1) to obtain yellow solid 22c (78 mg, yield 67%). ESI-MS (m / z): 416.5 [M+H]+. + .

[0325] Fourth step: Compound 22c (78 mg, 187 umol) was dissolved in dichloromethane (5 mL), and 4M hydrochloric acid dioxane solution (0.1 mL) was added at 0 °C and stirred overnight. TLC monitoring showed that the reaction was completed. Directly rotary evaporation to obtain white solid 22d (45 mg, yield 76%). ESI-MS (m / z): 316.5 [M+H]+. + .

[0326] Fifth step: Compound 22d (40 mg, 114 umol), compound 13d (25 mg, 88.4 umol) and p-toluenesulfonic acid monohydrate (1.52 mg, 8.82 umol) were dissolved in n-butanol (3 mL) and reacted at 160 °C under microwave for 3 hours. LCMS monitoring showed that the reaction was completed. The reaction solution was purified by reverse phase preparative HPLC to obtain white solid 22 (5 mg, yield 10%). ESI-MS (m / z): 562.4 [M+H]+. + ; 1H NMR (500 MHz, DMSO-d6) δ 7.12 (d, J = 7.9 Hz, 1H), 6.95 (d, J = 7.9 Hz, 1H), 6.50 (br s, 1H), 5.07 (t, J = 5.4 Hz, 1H), 4.28 - 4.16 (m, 1H), 3.93 (d, J = 5.1 Hz, 2H), 3.76 - 3.68 (m, 2H), 3.65 - 3.53 (m, 6H), 2.98 (s, 3H), 2.51 - 2.45 (m, 2H), 2.43 - 2.35 (m, 3H), 1.92 - 1.68 (m, 8H), 1.33 (s, 3H).

[0327] Example 23

[0328] (S)-5-(hydroxymethyl)-4,5-dimethyl-2-(((1r,3S)-3-(3,4,5-trifluorophenoxy)cyclobutyl)amino)- 4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0329]

[0330] Compound 23 was obtained by replacing 12e in the fourth step of Example 13 with intermediate 1e, using similar methods and reaction procedures. ESI-MS (m / z): 464.5 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) δ 6.90 - 6.80 (m, 2H), 6.71 (d, J = 6.9 Hz, 1H), 5.07 (d, J = 5.0 Hz, 1H), 4.87 - 4.78 (m, 1H), 4.45 - 4.33 (m, 1H), 3.78 - 3.68 (m, 2H), 3.66 - 3.58 (m, 1H), 3.57 - 3.50 (m, 1H), 2.97 (s, 3H), 2.52 - 2.37 (m, 4H), 2.36 - 2.28 (m, 2H), 1.90 - 1.75 (m, 2H), 1.33 (s, 3H).

[0331] Example 24

[0332] (S)-5-(hydroxymethyl)-4-methyl-2-(((1r,3S)-3-(3,4,5-trifluorophenoxy)cyclobutyl)amino)- 4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0333]

[0334] Compound 24 was prepared from the following steps:

[0335]

[0336] First Step: Compound 16c (500 mg, 1.86 mmol) and iodomethane (343 mg, 2.42 mmol) were dissolved in acetonitrile (10 mL), cesium carbonate (1.21 g, 372 mmol) was added, and the reaction was carried out at 50 °C for 2 hours. The reaction was monitored by LCMS. The reaction solution was diluted with dichloromethane, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to obtain white solid 24a (350 mg, yield 66%). ESI-MS (m / z): 283.3 [M+H] + .

[0337] Second Step: 24a (250 mg, 0.88 mmol) was dissolved in dichloromethane (2 mL), and boron tribromide (2.21 g, 8.84 mmol, 0.85 mL) was added dropwise at 0 °C. After the addition was completed, the reaction was carried out at 0 °C for two hours. The reaction was monitored by LCMS. The reaction solution was carefully quenched with sodium bicarbonate solution, extracted with dichloromethane, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (dichloromethane / methanol = 10 / 1) to obtain white solid 24b (100 mg, yield 42%). ESI-MS (m / z): 269.3 [M+H] + .

[0338] Third Step: Compound 24b (30 mg, 111 umol), compound 1e (42 mg, 167 umol) and p-toluenesulfonic acid monohydrate (1.92 mg, 11.1 umol) were dissolved in n-butanol (3 mL) and the reaction was carried out at 160 °C in a microwave for 3 hours. The reaction was monitored by LCMS. The reaction solution was purified by reverse phase preparative HPLC to obtain white solid 24 (16 mg, yield 32%). ESI-MS (m / z): 450.3 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) δ 6.91-6.79 (m, 2H), 6.76-6.69 (m, 1H), 5.01-4.95 (m, 1H), 4.87-4.81 (m, 1H), 4.45-4.33 (m, 1H), 4.16-3.94 (m, 2H), 3.80-3.63 (m, 2H), 2.97 (s, 3H), 2.51-2.44 (m, 2H), 2.41-2.35 (m, 2H), 1.95-1.85 (m, 1H), 1.81-1.69 (m, 1H).

[0339] Example 25

[0340] (S)-4,5-dimethyl-2-(((1r,3S)-3-((2-methyl-6-(trifluoromethyl)pyridin-3-yl)oxy)cyclobutyl)amino)- 4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0341]

[0342] Compound 25 was obtained by replacing intermediate 14d for 2a in the first step of Example 2, using similar procedures and reaction steps. ESI-MS (m / z): 463.5 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) δ 7.66 (d, J = 8.5 Hz, 1H), 7.25 (d, J = 8.5 Hz, 1H), 6.88 (d, J = 6.8 Hz, 1H), 4.97 (br s, 1H), 4.50-4.36 (m, 1H), 4.11 (q, J = 6.8 Hz, 1H), 4.04-3.98 (m, 1H), 3.35-3.29 (m, 2H), 2.94 (s, 3H), 2.60-2.30 (m, 9H), 2.00-1.88 (m, 1H), 1.85-1.75 (m, 1H), 1.23 (d, J = 6.6 Hz, 3H).

[0343] Example 26

[0344] (S)-4,5-dimethyl-2-(((1r,3S)-3-((4-methyl-2-(trifluoromethyl)pyrimidin-5-yl)oxy)cyclobutyl)amino)- 4,5,9,10-tetrahydro-6H,8H-pyrido[3,2,1-de]pteridin-6-one

[0345]

[0346] Compound 26 was prepared by the following steps:

[0347]

[0348] First step: Ethyl formate (3.17 g, 42.73 mmol) and sodium ethoxide (3.49 g, 50.50 mmol) were added to dry tetrahydrofuran (140 mL) successively. After the addition of compound 26a (7 g, 38.85 mmol) at 5-10 °C, the reaction was warmed to 50 °C and stirred for 2 hours. HPLC monitoring showed that the starting material was consumed. The tetrahydrofuran was evaporated under reduced pressure to give yellow oil 26b, which was used directly in the next step.

[0349] Second step: add the oil 26b obtained in the previous step into 150 ml of anhydrous ethanol, stir to dissolve at room temperature, add trifluoroacetamidine (4.35 g, 33.01 mmol, purity 85%) dropwise, and stir at 30°C for 5 hours, then warm to 80°C and continue to stir for 2 hours. HPLC monitoring shows that the raw material disappears. After cooling the reaction solution, about 100 ml of ethanol is distilled off under reduced pressure. The remaining residue is added to 300 ml of ice water, and the pH is adjusted to 3 with concentrated hydrochloric acid. Stir for 0.5 hours, filter, and dry the filter cake to obtain yellow solid compound 26c (4.37 g, yield 41%, purity 99%). ESI-MS (m / z): 271.4 [M+H] + .

[0350] Third step: add compound 26c (4.0 g, 14.80 mmol) to 60 ml of acetonitrile, and add phosphorus oxychloride (6.81 g, 44.41 mmol) dropwise. After the addition is complete, stir for 10 minutes, warm to 80°C, and stir for 2 hours. HPLC monitoring shows that the raw material is completely converted. Remove the acetonitrile under reduced pressure, and add the residual liquid to 200 mL of ice water. Stir for 0.5 hours, filter, and obtain yellow solid 26d (3.9 g, yield 86%, purity 95%).

[0351] Fourth step: add compound 26d (2.0 g, 6.93 mmol), trimethylboroxine (2.61 g, 20.79 mmol), palladium acetate (155 mg, 0.69 mol), and potassium phosphate (2.94 g, 13.86 mmol) to 1,4-dioxane (150 mL) in sequence, add water (15 mL), and stir at 90°C for 17 hours under nitrogen protection. HPLC monitoring shows that the raw material is completely converted, and the filtrate is concentrated. Purify the residue by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 9) to obtain white solid 26e (1.86 g, yield 50%, purity 99%). ESI-MS (m / z): 269.1 [M+H] + .

[0352] Fifth step: dissolve compound 26e (280 mg, 1.04 mmol) in methanol (10 mL), add palladium-carbon (13 mg, 104 umol), and replace the reaction system with hydrogen. React for 5 hours, and TLC monitoring shows that the reaction is complete. Filter the reaction solution with diatomite, concentrate the filtrate, and obtain yellow liquid 26f (170 mg, yield 91%). ESI-MS (m / z): 177.5 [M-H] - .

[0353] Step 6: Compound 26f (120 mg, 673 umol) and compound 1b (268 mg, 1.01 mmol) and cesium carbonate (548 mg, 1.68 mmol) were dissolved in N,N-dimethylformamide (5 mL) and stirred at 90 °C overnight. TLC monitored the end of the reaction, the reaction liquid was diluted with water, extracted with ethyl acetate, washed with saturated brine, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1) to obtain yellow solid 26g (148 mg, yield 62%). ESI-MS (m / z): 348.5 [M+H] + .

[0354] Step 7: Compound 26g (148 mg, 340 umol) was dissolved in dichloromethane (5 mL), 4M hydrochloric acid (0.4 mL) was added at 0 °C and stirred overnight. TLC monitored the end of the reaction, directly rotary dried to obtain white solid 26h (84 mg, yield 86%). ESI-MS (m / z): 284.4 [M+H] + .

[0355] Step 8: Compound 26h (30 mg, 118 umol), compound 1i (35 mg, 142 umol) and p-toluenesulfonic acid monohydrate (2 mg, 11.8 umol) were dissolved in n-butanol (3 mL) and reacted at 160 °C for 3 hours under microwave. LCMS monitored the end of the reaction. The reaction liquid was purified by reverse phase preparative HPLC to obtain white solid 26 (14 mg, yield 25%). ESI-MS (m / z): 464.5 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) δ 8.34 (s, 1H), 6.87 (d, J = 6.9 Hz, 1H), 5.19-5.08 (m, 1H), 4.52-4.41 (m, 1H), 4.12 (q, J = 6.9 Hz, 1H), 4.08-3.99 (m, 1H), 3.29-3.24 (m, 2H), 2.95 (s, 3H), 2.58-2.41 (m, 9H), 2.00-1.90 (m, 1H), 1.86-1.73 (m, 1H), 1.23 (d, J = 6.8 Hz, 3H).

[0356] According to the above described synthetic routes and the synthesis methods of intermediates, the following examples can be obtained.

[0357]

[0358]

[0359] Biological screening and results of Wnt pathway inhibitors

[0360] Test Example 1: Construction of Colo205-LUC-TCF / LEF-M1 reporter cell line

[0361] Colo205 cell line (Cell Bank of China Academy of Sciences, Cat#TCHu102) was purchased from Cell Bank of China Academy of Sciences. After expansion and subculture, the cells were in exponential growth phase. The cells were transfected with TCF / LEF transcription factor driven luciferase reporter plasmid (Promega) by lipo3000 liposome transfection method. The plasmid contains a resistance gene, which can be used for resistance screening. The transfection was carried out in a 10 cm culture dish, and the conventional complete culture medium without resistance was used. After 2 days, the medium with resistance was replaced and the culture was continued. Then the resistance medium was replaced every 2 days, and the suspended cells were discarded. The original culture medium was centrifuged to remove cells and debris, and was retained as an adaptive medium. When the cells grew to cover the culture dish, the cells were digested, counted, and subcultured into a 96-well plate, so that the number of cells contained in each well was averaged to be 1.5 per well. The adaptive medium was used during subculture. The remaining cells were frozen. After subculture for 4 hours, the cells were allowed to adhere, and then the number of cells in each well was observed under a microscope. Wells with only one cell per well were labeled as single clone wells. Then the normal culture was carried out, the medium was replaced every 2 days, and the observation was carried out. The single clone cells that continued to grow were labeled twice, and the normal resistance medium was replaced. When the cells in the single clone wells grew to cover the 96-well plate, they were digested and subcultured into a 24-well plate. When the 24-well plate was full, it was subcultured into a 96-well plate and a 6-well plate. The cells in the 96-well plate were subcultured into at least 6 wells, and 3 of them were treated with a known Wnt inhibitor, and the other 3 were not treated. After 24 hours, the 96-well plate cells were added with a luciferase detection reagent to detect the fluorescence intensity. The cell line with fluorescence expression without treatment and reduced fluorescence after inhibition was selected for further culture. The Colo205-LUC-TCF / LEF-M1 cell line is one of the cell lines selected by the above screening. Its growth curve, cell morphology, and cell growth state are similar to those of the original Colo205 cells. The ratio of fluorescence signals with and without inhibitor treatment is relatively large among all cell lines, and the ratio can reach 4-5 times after 4 hours of inhibition, which is completely suitable for the later screening of Wnt inhibitors.

[0362] Test Example 2: Detection of the inhibition ability of compounds on Colo205-LUC-TCF / LEF M1 reporter cell line

[0363] Colo205-LUC-TCF / LEF M1 cell line is a report tool cell stably transfected with pGL4.49-LUC2-TCF / LEF vector, and the β-catenin Wnt pathway is continuously activated. After adding an inhibitor, the Wnt pathway is inhibited, the expression amount of firefly luciferase regulated by TCF / LEF cis-element on the vector decreases, and after subsequently adding a detection substrate, the detected light signal correspondingly decreases, so as to detect the inhibition effect of the compound.

[0364] 100 μL of the compound with a maximum concentration of 20 μM was added to each well of a 96-well cell culture plate, and the compound concentration was diluted by 3 times in gradient. Then, 10000 stable reporter gene transfected colo205 cells and 100 μL of culture medium were inoculated into each well, and the corresponding treatment was performed as a positive and negative control well. The cells were placed in a 5% CO2 cell incubator and cultured at 37°C for 4 h. After 4 h, the culture medium was removed, 100 μL of reagent (Promega) containing the corresponding firefly luciferase substrate was added to each well, and the luciferase reporter gene activity was determined. The light intensity was read by SpectraMax in full wavelength mode. The light signal intensity of the cells treated only by DMSO was the positive control, and the light signal intensity of the cell-free well was the negative control. The IC 50 concentration of each compound was calculated. The colo 205 reporter gene detection data are summarized in Table 1 below.

[0365] Table 1 IC value of compound on Colo205-LUC-TCF / LEF reporter gene inhibition 50

[0366]

[0367]

[0368] Test Example 3: Proliferation inhibition test of compound on Wnt mutant cell lines (Colo205, DU4475, NCI-H929 and HepG2) and non-Wnt mutant cell lines (Hela and RKO)

[0369] The cell lines used in the test are Colo205, DU4475, NCI-H929 and HepG2 cell lines with continuously activated Wnt pathway and Wnt pathway dependent proliferation; and HE LA and RKO cell lines with normal Wnt pathway not activated and proliferation not dependent on Wnt pathway as control cell lines, so as to judge that the inhibition effect of the compound of the application on Wnt dependent proliferation is not caused by other non-specific toxicity.

[0370] ​Colo205, Du4475, NCI-H929, HepG2, HELA and RKO cell lines cultured in respective culture medium were treated in logarithmic growth phase, and after the cells were collected, a uniform cell suspension of known concentration was prepared, and then the cell suspension was added to a 96-well cell culture plate so that each well contained 1000 cells. It was placed in a 5% CO2 cell incubator at 37°C for 20-24h. The next day, the completely dissolved 3-fold gradient diluted compound was added to each cell culture well, and the final highest concentration in the cell culture well was 20μM, and it was continued to be cultured for 96h. Promega's cell viability detection test was used for detection, and the more the cell proliferation, the stronger the final signal strength. The detection instrument was SpectraMax, full wavelength mode. Only the DMSO added well was used as the positive control well, and the well without inoculating cells was used as the negative control well, and the IC 50 value of each compound for the proliferation inhibition of Wnt continuously activated or proliferation dependent cells, and the IC 50 value of the proliferation inhibition of Wnt non-activated or proliferation independent cells were calculated, the inhibitory effect of the compound on the Wnt pathway and the toxic effect on normal cells were evaluated. The results are shown in Table 2 below.

[0371] Table 2 IC 50 value of the proliferation inhibition of Wnt mutant cell lines

[0372]

[0373]

[0374] The above results show that the compound of the present application has significant inhibitory activity on mutant cell lines Colo205, DU4475, NCI-H929 and HepG2, and basically has no significant inhibitory activity on Hela and RKO cell lines, which shows that the compound of the present application has significant and selective Wnt pathway inhibition.

Claims

1. A compound having the following structure:

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

3. Use of the compound of claim 1 or a pharmaceutically acceptable salt thereof and the pharmaceutical composition of claim 2 in the manufacture of a medicament for the prevention and / or treatment of cancer, an inflammatory disease.

4. Use of the compound of claim 1 or a pharmaceutically acceptable salt thereof and the pharmaceutical composition of claim 2 in the manufacture of a medicament for the prevention and / or treatment of a tumor, an autoimmune disease.

5. Use of the compound of claim 1 or a pharmaceutically acceptable salt thereof and the pharmaceutical composition of claim 2 in the manufacture of a medicament for the prevention and / or treatment of an immune-mediated disease.

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