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

TECHNICAL FIELD

[0001] The present application relates to a heterocyclic compound, in particular to a highly active Wnt pathway inhibitor and uses thereof. BACKGROUND

[0002] Wnt / β-catenin signaling pathway is a conserved pathway in biological evolution. In normal somatic cells, β-catenin only plays a role in maintaining the adhesion of homotypic cells and preventing cell movement as a cytoskeletal protein at the cell membrane in the form of a 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, etc., thereby initiating the degradation of β-catenin by the ubiquitin system through the proteasome pathway, and maintaining the 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, and stabilize the free state of β-catenin protein in the cytoplasm. The stable accumulation of β-catenin in the cytoplasm enters the nucleus to bind to the LEF / TCF transcription factor family, and initiates the transcription of downstream target genes (such as c-myc, c-jun, Cyclin D1, etc.). Overactivation of the Wnt / β-catenin signaling pathway is closely related to the occurrence of various cancers (including colon cancer, gastric cancer, breast cancer, etc.). For example, abnormal activation of the Wnt classic signaling pathway and nuclear accumulation of β-catenin protein are widely present in colorectal cancer, and inhibition of Wnt signaling pathway activity can inhibit the proliferation of cancer such as colon cancer. Mutations in APC are present 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 and β-catenin mutation itself can also cause intracellular aggregation of β-catenin and activation of the Wnt / β-catenin pathway.

[0003] Although it is known that inhibition of the Wnt signaling pathway can effectively prevent and / or treat cancer, tumor, inflammatory disease, autoimmune disease and immune-mediated disease, there is still a lack of satisfactory effective Wnt pathway inhibitor compounds in the prior art. Therefore, it is necessary to study effective Wnt pathway inhibitor compounds in the prior art. SUMMARY

[0004] In one aspect, the present application provides a compound having Formula (I) or a pharmaceutically acceptable salt, isotopically enriched derivative, stereoisomer thereof, which inhibits Wnt pathway activity:

[0005]

[0006] wherein,

[0007] R1, R2 each independently represents hydrogen, (C1-C6)alkyl, halo(C1-C6)alkyl, (C3-C8)cycloalkyl, halo(C3-C8)cycloalkyl, 4-8 membered heterocycloalkyl, halo 4-8 membered heterocycloalkyl, -(C1-C6)alkyleneOR a , -halo(C1-C6)alkyleneOR a , -(C1-C6)alkyleneSR a , -halo(C1-C6)alkyleneSR a , or R1, R2 together with the carbon atom to which they are attached form a 3-8 membered ring which can optionally contain 0, 1, 2 or 3 heteroatoms selected from N, O and S;

[0008] R3 represents (C1-C6)alkyl, halo(C1-C6)alkyl, (C3-C8)cycloalkyl, halo(C3-C8)cycloalkyl, or R3 and R1 or R2 together form a 4-7 membered ring which can optionally contain 0 or 1 heteroatoms selected from O and S;

[0009] W6 represents CR6 or N;

[0010] R6 each independently represents hydrogen, halogen, cyano, (C1-C3)alkyl, halo(C1-C3)alkyl;

[0011] Cy represents a 5-12 membered heteroaromatic ring which optionally contains 1, 2, 3 or 4 heteroatoms each independently selected from N, O and S, and which is unsubstituted or substituted with hydrogen at the ortho position to the -O- attachment position;

[0012] R1’ represents hydrogen, halogen, (C1-C6)alkyl, halo(C1-C6)alkyl, (C3-C8)cycloalkyl, halo(C3-C8)cycloalkyl, (C3-C8)heterocycloalkyl, halo(C3-C8)heterocycloalkyl, -OR a , -haloOR a , -SR a , -haloSR a ;

[0013] R2' represents hydrogen, halogen, (Ci-C6)alkyl, halo(Ci-C6)alkyl, (C3-C8)cycloalkyl, halo(C3-C8)cycloalkyl, (C3-C8)heterocycloalkyl, halo(C3-C8)heterocycloalkyl, -OR a , -haloOR a , -SR a , -haloSR a ;

[0014] R3' represents hydrogen, halogen, (Ci-C6)alkyl, halo(Ci-C6)alkyl, (C3-C8)cycloalkyl, halo(C3-C8)cycloalkyl, (C3-C8)heterocycloalkyl, halo(C3-C8)heterocycloalkyl, -OR a , -haloOR a , -SR a , -haloSR a ;

[0015] m each independently represents 0, 1 or 2;

[0016] R a each independently represents hydrogen, (Ci-C6)alkyl, halo(Ci-C6)alkyl, (C3-C8)cycloalkyl, halo(C3-C8)cycloalkyl.

[0017] In one aspect, Cy in the structure of formula (I) is a 5-membered heteroaromatic ring or a 6-membered heteroaromatic ring. More preferably, Cy is pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, pyrazolyl, imidazolyl or pyrrolyl.

[0018] In yet another aspect, the compound of formula (I) is as shown in formula (II):

[0019]

[0020] wherein,

[0021] the dotted circle indicates that the ring in which it is located is aromatic;

[0022] W1 represents CR1' or N;

[0023] W2 represents CR2' or N;

[0024] W3 represents CR3' or N;

[0025] W4 represents CH or N;

[0026] W5 represents CH or N;

[0027] R1, R2, R3, R6, W6, R1', R2', R3', R a, m is as described in claim 1. More preferably, the compound of formula (II) is as shown below:

[0028]

[0029] In another aspect, the compound of formula (I) is as shown in formula (III):

[0030]

[0031] wherein,

[0032] the dotted line circle indicates that the ring in which it is located is an aromatic ring;

[0033] W1' represents CH, N or NH;

[0034] W2' represents CR2', N or NR2';

[0035] W3' represents CR3', N or NR3';

[0036] W4' represents CH, N or NH;

[0037] R1, R2, R3, R6, W6, R1', R2', R3', R a , m is as described in claim 1.

[0038] More preferably, the compound of formula (III) is as shown below:

[0039]

[0040] In one aspect, R1, R2 each independently represents hydrogen, (C1-C6)alkyl, halo(C1-C6)alkyl, (C3-C8)cycloalkyl, halo(C3-C8)cycloalkyl, 4-8 membered heterocycloalkyl, halo 4-8 membered heterocycloalkyl, -(C1-C6)alkyleneOR a , -halo(C1-C6)alkyleneOR a ; preferably hydrogen, (C1-C6)alkyl, halo(C1-C6)alkyl.

[0041] In one aspect, R3 is (C1-C6)alkyl.

[0042] In one aspect, W6 is CR6.

[0043] In one aspect, R6 is hydrogen.

[0044] In one aspect, R1' represents hydrogen, halogen, (C1-C6)alkyl, halo(C1-C6)alkyl, (C3-C8)cycloalkyl, halo(C3-C8)cycloalkyl. More preferably, R1' represents halo(C1-C6)alkyl.

[0045] In one aspect, R2' represents hydrogen, halogen, (Ci-C6)alkyl, halo(Ci-C6)alkyl, (C3-C8)cycloalkyl, halo(C3-C8)cycloalkyl. More preferably, R2' represents hydrogen or halogen.

[0046] In one aspect, R3' represents halogen, (Ci-C6)alkyl, halo(Ci-C6)alkyl, (C3-C8)cycloalkyl, halo(C3-C8)cycloalkyl, -OR a , -haloOR a , -SR a , -haloSR a . Preferably, R3' represents halogen, (Ci-C6)alkyl, halo(Ci-C6)alkyl, (C3-C8)cycloalkyl, halo(C3-C8)cycloalkyl. More preferably, R3' represents halo(Ci-C6)alkyl or halo(C3-C8)cycloalkyl.

[0047] In some preferred aspects, the present application provides a compound having the following structure:

[0048]

[0049]

[0050] Further, the present application also provides a pharmaceutical composition comprising a compound of the present application or a pharmaceutically acceptable salt, isotope derivative or stereoisomer thereof.

[0051] Further, the present application also provides the use of a compound of the present application or a pharmaceutically acceptable salt, isotope derivative or stereoisomer thereof, or a pharmaceutical composition of the present application, for 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 is generally meant to also encompass stereoisomers, diastereomers, enantiomers, racemic mixtures and isotope derivatives thereof.

[0052] 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, and therefore, it is particularly noted that, herein, when referring to a compound of the structure of Formula (I) to Formula (III), it is generally meant to also encompass a pharmaceutically acceptable salt thereof, and further to encompass a solvate and a hydrate thereof.

[0053] Similarly, herein, when referring to a compound, it is generally meant to also encompass a prodrug, a metabolite and a nitroxide thereof.

[0054] 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.

[0055] 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.

[0056] 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.

[0057] 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.

[0058] 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.

[0059] 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 or reduced dosage requirements.

[0060] The present application also provides the use of a compound of the 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.

[0061] 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, which comprises a compound of the application as an active ingredient. The pharmaceutical composition can optionally comprise a pharmaceutically acceptable carrier.

[0062] 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, which comprises administering a compound of the application to a mammal in need thereof.

[0063] Representative examples of inflammatory diseases, autoimmune diseases, and immune-mediated diseases may include, but are not limited to, arthritis, rheumatoid arthritis, spondyloarthritis, gouty arthritis, osteoarthritis, juvenile arthritis, other inflammatory joint 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 inflammatory lung disease, chronic obstructive pulmonary disease (COPD), cardiovascular diseases, atherosclerosis, myocardial infarction, congestive heart failure, myocardial ischemia-reperfusion injury, inflammatory bowel disease, Crohn's disease, ulcerative colitis, irritable bowel syndrome, asthma, Sjögren's syndrome, and autoimmune thyroid diseases. Diseases, urticaria (rubella), multiple sclerosis, scleroderma, organ transplant rejection, xenotransplantation, idiopathic thrombocytopenic purpura (ITP), Parkinson's disease, Alzheimer's disease, diabetes-related diseases, inflammation, pelvic inflammatory disease, allergic rhinitis, allergic bronchitis, allergic sinusitis, leukemia, lymphoma, B-cell lymphoma, T-cell lymphoma, myeloma, acute lymphoblastic leukemia (ALL), chronic lymphoblastic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), hairy cell leukemia, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, myelodysplastic syndrome (MDS), myeloproliferative neoplasm (MPN), diffuse large B-cell lymphoma, and follicular lymphoma.

[0064] Representative examples of cancers or tumors may include, but are not limited to, skin cancer, bladder cancer, ovarian cancer, breast cancer, stomach cancer, pancreatic cancer, prostate cancer, colon cancer, lung cancer, bone cancer, brain cancer, neurocytoma, rectal cancer, colon cancer, familial adenomatous polyposis, hereditary nonpolyposis colorectal cancer, esophageal cancer, lip cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, stomach cancer, adenocarcinoma, medullary thyroid carcinoma, papillary thyroid carcinoma, kidney cancer, renal parenchymal carcinoma, ovarian cancer, cervical cancer, uterine cancer, endometrial cancer, choriocarcinoma, pancreatic cancer, prostate cancer, testicular cancer, urinary tract cancer, melanoma, brain tumors such as glioblastoma, astrocytoma, meningioma, medulloblastoma, and peripheral thyroid cancer. Neuroectodermal tumors, Hodgkin's lymphoma, non-Hodgkin's lymphoma, Burkitt's lymphoma, acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), adult T-cell leukemia lymphoma, diffuse large B-cell lymphoma (DLBCL), hepatocellular carcinoma, gallbladder cancer, bronchial carcinoma, small cell lung cancer, non-small cell lung cancer, multiple myeloma, basal cell tumor, teratoma, retinoblastoma, choroidal melanoma, seminoma, rhabdomyosarcoma, craniopharyngioma, osteosarcoma, chondrosarcoma, myoma, liposarcoma, fibrosarcoma, Ewing's sarcoma, or plasmacytoma.

[0065] 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.

[0066] 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.

[0067] 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.

[0068] 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.

[0069] 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.

[0070] In addition to this, 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.

[0071] 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.

[0072] 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.

[0073] Examples of carriers that can be 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.

[0074] Other features of the application will be apparent from consideration of the specification and the examples, given by way of illustration of the application and not intended to be limiting thereof, which follow.

[0075] 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 solvent used. One skilled in the art of organic synthesis will recognize the utility of alternative methods for the preparation of these compounds. The preferred method of preparation will depend on the availability of starting materials, and the desired synthetic route. BRIEF DESCRIPTION OF DRAWINGS

[0076] Figure 1 The effect of Compound 1 on tumor growth of human colon cancer cell Colo205 xenograft tumor is shown. DETAILED DESCRIPTION

[0077] TERMINOLOGY

[0078] The terms, as used in the present application, including the specification and claims, are defined as follows. If not otherwise specified, the conventional methods of mass spectrometry, nuclear magnetic, HPLC, protein chemistry, biochemistry, recombinant DNA techniques and pharmacology are used. In the present application, if not otherwise specified, "or" or "and" means "and / or".

[0079] In the specification and claims, a given chemical formula or name shall cover all of its stereoisomers and optical isomers and the racemates in which such isomers are present. Unless otherwise indicated, all chiral (enantiomers and diastereomers) and racemic forms are within the scope of the present invention. Multiple geometric isomers of C═C double bonds, C═N double bonds, ring systems, etc. may also be present in the compounds, and all such stable isomers are covered within 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 separate isomer 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 to be part of the present invention. When preparing enantiomeric or diastereomeric products, they can be separated by conventional methods (e.g., by chromatography or fractional crystallization). Depending on the method conditions, the end products of the present invention are obtained in free (neutral) or salt form. Both the free form and the salts of these end products are within the scope of the present invention. If desired, 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 the 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. It should be understood that all tautomeric forms that can exist are included within the present invention.

[0080] Unless otherwise defined, the definitions of the substituents of the present invention are each independent and not 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 selecting one definition in one substituent, it does not mean that this R a (or R a ’) has the same definition in other substituents. More specifically, for example (by way of illustration only and not limitation), for NR a R a ’, when the definition of R a (or R a ’) is selected from hydrogen, it does not mean that in -C(O)-NR a R a ’, R a (or R a ’) must necessarily be hydrogen. In another aspect, when there is more than one R a (or Ra ) are each independently as defined above. For example, in the substituent -(CR a (Or R a ) are each independently as defined above. For example, in the substituent -(CR a R a’ ) m -O-(CR a R a’ ) n - In the substituent -O-(CR a (Or R a ) are each independently as defined above. For example, in the substituent -O-(CR

[0081] Unless otherwise defined, when a substituent is designated as "optionally substituted" the substituent is selected from, for example, alkyl, cycloalkyl, aryl, heterocyclyl, halo, hydroxy, alkoxy, oxo, alkanoyl, aryloxy, alkanoyloxy, amino, alkylamino, arylamino, arylalkylamino, disubstituted amino (wherein the 2 amino substituents are selected from alkyl, aryl, or arylalkyl), alkanoylamino, aroylamino, aralkanoylamino, substituted alkanoylamino, substituted arylamino, substituted aralkanoylamino, thio, alkylthio, arylthio, arylalkylthio, arylthiocarbonyl, arylalkylthiocarbonyl, alkylsulfonyl, arylsulfonyl, arylalkylsulfonyl, aminosulfonyl such as -SO2NH2, substituted sulfonamido, nitro, cyano, carboxy, carbamoyl such as -CONH2, substituted carbamoyl such as -CONHalkyl, -CONHaryl, -CONHarylalkyl, or where the nitrogen has two substituents selected from alkyl, aryl, or arylalkyl, alkoxycarbonyl, aryl, substituted aryl, guanidino, heterocyclyl, such as indolyl, imidazolyl, furanyl, thienyl, thiazolyl, pyrrolidinyl, pyridyl, pyrimidinyl, pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, homopiperazinyl, and substituted heterocyclyl.

[0082] The term "alkyl" or "alkylene" as used herein is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms. For example, "Ci-C6alkyl" denotes alkyl groups having from 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, t-butyl) and pentyl (e.g., n-pentyl, isopentyl, neopentyl). In the present context, alkyl is preferably alkyl having 1 to 6, more preferably 1 to 4 carbon atoms.

[0083] 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.

[0084] 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.

[0085] 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-.

[0086] The term "carbonyl" refers to the organic functional group (C=0) connected by a double bond to two atoms, one of which is carbon.

[0087] 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.

[0088] 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.

[0089] 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.

[0090] "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.

[0091] "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-.

[0092] 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.

[0093] 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.

[0094] In the present disclosure, one or more halogen can each independently be selected from fluorine, chlorine, bromine, and iodine.

[0095] 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 heterocyclic groups described herein can be substituted on a carbon or a nitrogen atom if the resulting compound is stable. The nitrogens in the heterocyclic rings can optionally be quaternized. Preferably, when the total number of S and O atoms in the heterocyclic ring exceeds 1, 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 1. 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.

[0096] 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.

[0097] 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.

[0098] The term "spirocycloalkyl" as used herein refers to polycyclic hydrocarbons sharing one carbon atom (called spiro atom) between single rings.

[0099] 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.

[0100] The term "spiroheterocyclyl" as used herein refers to polycyclic hydrocarbons sharing one carbon atom (called spiro atom) between single rings, at least one of which is a heteroatom selected from O, N and S.

[0101] 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).

[0102] In cases where 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.

[0103] 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.

[0104] 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.

[0105] 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 symptoms of a disease, condition, or disorder, or to enhance normal physiological function, as compared to an untreated subject. 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.

[0106] The term "treatment" as used herein includes any effect that relieves, reduces, modulates, ameliorates, or eliminates a condition, disease, disorder, etc., or its symptoms.

[0107] 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.

[0108] 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;

[0109] 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.

[0110] Specific Pharmaceutical and Medical Terms

[0111] 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.

[0112] 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).

[0113] 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.

[0114] 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.

[0115] 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.

[0116] The terms "kit" and "product package" are synonymous.

[0117] 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.

[0118] 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 symptom, such as controlling the progression of a disease or condition; abating a disease or symptom; causing regression of a disease or symptom; relieving a complication caused by a disease or symptom, or preventing and / or treating an indication caused by a disease or symptom.

[0119] As used herein, an improvement in a disease, symptom, or condition, with respect to a compound or pharmaceutical composition, means, inter alia, that the severity of the disease, symptom, or condition is improved, the onset of the disease, symptom, or condition is delayed, the progression of the disease, symptom, or condition is slowed, or the duration of the disease, symptom, or condition is lessened. The improvement can be attributed to or associated with the administration, whether fixed or contingent, whether continuous or intermittent.

[0120] Routes of administration

[0121] 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 illustration, intramuscular, subcutaneous, intravenous, intramedullary, intraventricular, intraperitoneal, intralymphatic, and intranasal.

[0122] 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.

[0123] Pharmaceutical compositions and dosages

[0124] 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 uses, e.g., orally, such as tablets, pills, powders, granules, elixirs, tinctures, suspensions including nano-, micro-, 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, for example, 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.

[0125] 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.

[0126] 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.

[0127] 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.

[0128] 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 the like, and consistent with conventional pharmaceutical practices.

[0129] The dosage forms (pharmaceutical compositions) suitable for administration can contain from about 1 milligram to about 2000 milligrams of the active ingredient per dosage unit. In these pharmaceutical compositions the active ingredient will ordinarily be present in an amount of about 0.1-95% by weight based on the total weight of the pharmaceutical composition.

[0130] 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.

[0131] A typical injectable formulation can be prepared as follows: at least one compound of the present application (250 mg) is placed into a vial, lyophilized from sterile water, and the container sealed. For use, the contents of the vial is reconstituted with 2 mL of sterile water to give an injectable formulation.

[0132] 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).

[0133] 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.

[0134] Actual dosage levels of the active ingredients in the pharmaceutical compositions of the present application can be varied so as to obtain an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

[0135] 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.

[0136] 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. Generally, oral, intravenous, intracerebroventricular, and subcutaneous doses of the compounds of the application for patients will range from about 0.01 to about 50 mg / kg body weight per day. If desired, the effective daily dose can be divided into two, three, four, five, six, or more sub-doses that are administered separately at appropriate intervals during the day, optionally, in unit dosage forms. In certain aspects of the application, dosing is once daily.

[0137] While it is possible for a compound of the present application to be administered alone, it is preferable to administer the compound as a pharmaceutical formulation (composition).

[0138] Kits / Products Packets

[0139] Kits / Products Packets 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 of the methods described above. Suitable containers include bottles, tubes, syringes, and test tubes, among others. The containers are made of materials such as glass or plastic. Suitable containers include bottles, tubes, syringes, and test tubes, among others. The containers are made of materials such as glass or plastic.

[0140] For example, a container can hold one or more compounds described herein, which can be present in unit dosage form, and other ingredients described herein. A container can have a sterile access port (for example the container 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, labels, or packaging that describe manufacturing and / or use of the kits.

[0141] A typical kit can include one or more containers into which one or more materials (such as reagents, which can be concentrated stocks, and / or devices) can be placed. These materials include, but are not limited to, buffers, diluents, filters, needles, syringes, wands, receptacles, vials, and / or test tubes, to which checklists and / or instructions can be attached, and which can be packaged together in an instructional booklet. The entire instruction set can be included.

[0142] Labels suitable for use in conjunction with the containers and reagents of the application include labels made of paper, plastic or metal. A label can be attached to a container by a variety of means, such as by crimping, molding or etching, or the label can be imprinted on the container. A label can also appear on a box, carton or other container that holds a plurality of containers. A label can indicate that the contents are useful for a particular therapeutic treatment. A label can also indicate directions for use of the contents, such as in the methods described above.

[0143] 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 otherwise.

[0144] The above features of the invention, or features of the embodiments, can be combined in any combination. All of the features disclosed 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 otherwise.

[0145] The application will be further described with reference to the following examples. It should be appreciated that these examples are for illustrative purposes only and are not meant to limit the scope of the application. Unless otherwise indicated, the methods of preparing the examples below generally follow conventional procedures or those described in the examples. Unless otherwise indicated, all percentages, ratios, proportions, or parts are by weight.

[0146] The units in the weight volume percentages in the present application are well known to those skilled in the art, for example, it means the weight (g) of solute in 100 ml of solution. Unless otherwise defined, all other technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the application belongs. In addition, methods and materials similar or equivalent to those described herein can be used in the practice of the present application. The preferred methods and materials described herein are illustrative only and not intended to be limiting.

[0147] Examples

[0148] General procedures

[0149] When not included in the preparation route, the starting materials and reagents used in the present application are known products, which 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 are not required to be further purified.

[0150] Room temperature means 20-30 °C.

[0151] Unless otherwise specified in the reaction examples, the reactions are carried out under nitrogen atmosphere. The nitrogen atmosphere means that the reaction flask is connected to a nitrogen balloon of about 1 L.

[0152] The hydrogenation reaction is usually vacuumed and filled with hydrogen, and the operation is repeated 3 times. The hydrogen atmosphere means that the reaction flask is connected to a hydrogen balloon of about 1 L.

[0153] Microwave reaction uses Initiator + microwave reactor.

[0154] The structure of the compounds 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), deuterated methanol (CD3OD), and the internal standard is tetramethylsilane (TMS). The following abbreviations are used for the multiplicity of NMR signals: s = singlet, br s = broad singlet, d = doublet, t = triplet, m = multiplet. The coupling constant is listed as J value, measured in Hz.

[0155] 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 San Tai (S-1000) silica gel pre-packed column. Pre-packed column. TLC silica gel plate is Yantai Yellow Sea HSGF254 or Qingdao GF254 silica gel plate, and the specification of TLC separation and purification product is 0.4mm-0.5mm.

[0156] The synthesis method of some intermediates in the application is as follows:

[0157] Intermediate 1

[0158]

[0159] Intermediate 1 is prepared by the following steps:

[0160]

[0161] Step 1: Compound Int-1a (5.0g, 28.09mmol), Int-1b (5.61g, 36.51mmol) and sodium bicarbonate (7.08g, 84.26mmol) are dissolved in ethanol (50mL) and water (5mL), and the reaction solution is heated to reflux overnight. After the reaction is monitored to end by LCMS, it is cooled to room temperature, suction filtered, the filter cake is washed with water, and then dried to obtain off-white solid Int-1 (5.0g, yield 78%). ESI-MS (m / z): 227.4 [M+H] + .

[0162] Intermediate 2

[0163]

[0164] Intermediate 2 is prepared by the following steps:

[0165]

[0166] Step 1: Sodium hydride (1.59g, 39.64mmol, content 60%) is added to a two-necked flask containing anhydrous tetrahydrofuran (10mL), and the flask is placed in an ice water bath. Compound Int-2a (5.0g, 26.43mmol) is dissolved in anhydrous tetrahydrofuran (30mL), and slowly added to the reaction solution. After 30 minutes, deuterated methyl iodide (4.02g, 27.75mmol) is slowly added, and after the addition is completed, the solution is slowly warmed to room temperature and stirred overnight. After the reaction is monitored to end by LCMS, it is cooled to 0°C, and saturated ammonium chloride aqueous solution is slowly added dropwise to quench the reaction. Ethyl acetate is extracted, and the organic phase is dried to obtain yellow oil Int-2b (5.0g, yield 91%). ESI-MS (m / z): 207.2 [M+H] + .

[0167] Second step: Int-2b (5.0 g, 24.24 mmol) was dissolved in methanol (50 mL), placed in an ice water bath, and sulfuric chloride (8.65 g, 72.73 mmol) was slowly added dropwise. After the addition was complete, the reaction was allowed to warm to room temperature and then to 60 °C overnight. After the reaction was completed as monitored by LCMS, the reaction was concentrated to give yellow oil Int-2c (3.0 g, yield 79%). ESI-MS (m / z): 157.2 [M+H] + .

[0168] Third step: Compound Int-1a (2.0 g, 11.23 mmol), Int-2c (3.17 g, 20.22 mmol) and sodium bicarbonate (2.83 g, 33.70 mmol) were dissolved in ethanol (20 mL) and water (2 mL), and the reaction was refluxed overnight. After the reaction was completed as monitored by LCMS, it was cooled to room temperature, suction filtered, and the filter cake was washed with water and then dried to give off-white solid Int-2 (1.4 g, yield 54%). ESI-MS (m / z): 230.3 [M+H] + .

[0169] Intermediate 3

[0170]

[0171] Intermediate 3 was prepared by the following steps:

[0172]

[0173] First step: Sodium hydride (237 mg, 5.90 mmol, 60% content) was added to a two-necked flask containing anhydrous tetrahydrofuran (5 mL), and the flask was placed in an ice water bath. Compound Int-3a (300 mg, 1.48 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL) and slowly added dropwise to the reaction. After 30 minutes, deuterated methyl iodide (472 mg, 2.95 mmol) was added dropwise, and the reaction was slowly warmed to room temperature and stirred overnight. After the reaction was completed as monitored by LCMS, it was cooled to 0 °C, and saturated aqueous ammonium chloride solution was slowly added dropwise to quench the reaction. The reaction was extracted with ethyl acetate, and the organic phase was dried to give yellow oil Int-3b (312 mg, yield 96%). ESI-MS (m / z): 221.6 [M+H] + .

[0174] Second step: Int-3b (312 mg, 1.42 mmol) was dissolved in methanol (5 mL) and placed in an ice water bath, and thionyl chloride (505 mg, 4.25 mmol) was added dropwise. After the addition was complete, the reaction was allowed to warm to room temperature and then to 70 °C overnight. After the reaction was complete as monitored by LCMS, the reaction was concentrated to give yellow oil Int-3c (180 mg, 84% yield). ESI-MS (m / z): 135.4 [M+H] + .

[0175] Third step: Compound Int-3c (180 mg, 1.05 mmol) and compound 4d (200 mg, 0.95 mmol) were dissolved in tetrahydrofuran (5 mL), and N,N-diisopropylpropylamine (371 mg, 2.88 mmol) was added. The reaction was stirred at room temperature for 8 hours. The reaction was complete as monitored by LCMS. The reaction was diluted with water and extracted with ethyl acetate. The reaction was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1) to give Int-3d (105 mg, 35% yield) as off-white solid. ESI-MS (m / z): 306.5 [M+H] + .

[0176] Fourth step: Compound Int-3d (105 mg, 343 umol) was dissolved in glacial acetic acid (2 mL), and iron powder (77 mg, 1.37 mmol) was added. The reaction was stirred at room temperature for 16 hours. The reaction was complete as monitored by LCMS. The residue was filtered off with celite and washed with ethyl acetate. The filtrate was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane / methanol = 10 / 1) to give compound Int-3 (81 mg, 96% yield). ESI-MS (m / z): 244.4 [M+H] + .

[0177] Intermediate 4

[0178]

[0179] Intermediate 4 was prepared by the following steps:

[0180]

[0181] First Step: Compound Int-4a (1.01 g, 5 mmol) and potassium hydroxide (842 mg, 15 mmol) were dissolved in anhydrous dimethyl sulfoxide (10 mL), then deuterated methyl iodide (1.59 g, 11 mmol) was added dropwise into the reaction solution. The reaction solution was stirred at room temperature for 8 hours. LCMS was used to monitor the completion of the reaction. The reaction solution was diluted with water, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude compound Int-4b (1.0 g, yield 84%). ESI-MS (m / z): 236.3 [M+H] + .

[0182] Second Step: Compound Int-4b (1.0 g, 4.25 mmol) was dissolved in a hydrochloric acid dioxane solution (4 M, 10 mL). The reaction solution was stirred at room temperature for 4 hours. LCMS was used to monitor the completion of the reaction. The reaction solution was concentrated under reduced pressure to obtain the crude compound Int-4c (720 mg). ESI-MS (m / z): 136.1 [M+H] + .

[0183] Third Step: Compound Int-4c (720 mg, 4.19 mmol) and compound Int-3d (872 mg, 4.19 mmol) were dissolved in tetrahydrofuran (10 mL), and N,N-diisopropylpropylamine (1.62 g, 12.57 mmol) was added. The reaction solution was stirred at room temperature for 8 hours. LCMS was used to monitor the completion of the reaction. The reaction solution was diluted with water, filtered, the filter cake was washed with water, and dried to obtain an off-white solid Int-4d (993 mg, yield 77% for two steps). ESI-MS (m / z): 307.2 [M+H] + .

[0184] Fourth Step: Compound Int-4d (306 mg, 1 mmol) and potassium carbonate (207 mg, 1.5 mmol) were dissolved in methanol (10 mL), and sodium hydrosulfite (871 mg, 5 mmol) was dissolved in water and added dropwise into the reaction solution. The reaction solution was stirred at room temperature for 15 minutes. LCMS was used to monitor the completion of the reaction. Hydrochloric acid dioxane solution (4 M, 1.25 mL) was added to the reaction solution. The reaction solution was continued to be stirred at room temperature for 8 hours. Ammonia methanol solution was added dropwise into the reaction solution, the pH of the reaction solution was adjusted to 10, the reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane / methanol = 10 / 1) to obtain compound Int-4 (30 mg, yield 13%). ESI-MS (m / z): 242.3 [M+H] + .

[0185] Intermediate 5

[0186]

[0187] Intermediate 5 was prepared from the following steps:

[0188]

[0189] Step 1: Compound Int-5a (500 mg, 3.86 mmol), 6-fluoro-nicotinonitrile Int-5b (518.39 mg, 4.25 mmol) and cesium carbonate (1.89 g, 5.79 mmol) were dissolved in acetonitrile (10 mL) and stirred at room temperature overnight. The reaction was monitored by LCMS. The reaction was diluted with ethyl acetate and washed with water and saturated brine successively. The organic phase was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1) to give yellow oil 5c (680 mg, yield 76%). ESI-MS (m / z): 232.4 [M+H] + .

[0190] Step 2: Compound Int-5c (680 mg, 2.94 mmol) and Raney Nickel (0.5 mL, aqueous suspension) were dissolved and dispersed in methanol (10 mL) and aqueous ammonia (1 mL). The reaction was stirred at room temperature overnight under hydrogen balloon. The reaction was monitored by LCMS. The reaction was diluted with methanol and filtered through celite. The organic phase was concentrated and purified by silica gel column chromatography (dichloromethane / methanol = 10 / 1) to give yellow oil Int-5 (220 mg, yield 31%). ESI-MS (m / z): 236.4 [M+H] + .

[0191] Intermediate 6

[0192]

[0193] Intermediate 6 was prepared from the following steps:

[0194]

[0195] Step 1: Compound Int-1a (500 mg, 2.81 mmol) and compound Int-6a (602 mg, 3.65 mmol) were dissolved in ethanol (20 mL) and water (2 mL), followed by the addition of sodium bicarbonate (707 mg, 8.34 mmol). The temperature of the reaction was raised to 80 °C and stirred for 16 hours. After the reaction was cooled to room temperature, water (12 mL) was added to the reaction. The reaction was filtered to give off-white solid Int-6 (385 mg, yield 57%). ESI-MS (m / z): 239.4 [M+H] + .

[0196] Intermediate 7

[0197]

[0198] Intermediate 7 was prepared by the following steps:

[0199]

[0200] Step 1: 2-(trifluoromethyl)pyrimidin-5-ol Int-7a (1.0 g, 6.09 mmol), 6-fluoro- nicotinonitrile Int-5b (1.01 g, 7.31 mmol) and cesium carbonate (3.97 g, 12.19 mmol) were dissolved in DMF (10 mL) and stirred at 80 °C overnight. The reaction was monitored by LCMS. The reaction was diluted with ethyl acetate and washed with water and saturated brine successively. The organic phase was dried over anhydrous sodium sulfate, concentrated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1) to give Int-7b (1.3 g, yield 80%) as yellow oil.

[0201] Step 2: Compound Int-7b (1.3 g, 4.88 mmol) and Raney Nickel (0.5 mL, aqueous suspension) were dissolved and dispersed in methanol (10 mL) and ammonia (1 mL), and the reaction was stirred at room temperature overnight under hydrogen balloon. The reaction was monitored by LCMS. The reaction was diluted with methanol and filtered through celite. The organic phase was concentrated to give Int-7 (700 mg, yield 53%) as yellow oil. ESI-MS (m / z): 271.3 [M+H] + .

[0202] Intermediate 8

[0203]

[0204] Intermediate 8 was prepared by the following steps:

[0205]

[0206] Step 1: 6-(trifluoromethyl)pyridin-3-ol Int-8a (1.0 g, 6.13 mmol) was dissolved in dimethyl sulfoxide (10 mL) and cesium carbonate (2.0 g, 6.13 mmol) was added. After stirring at room temperature for 30 minutes, 2-fluoropyridine-5-carbaldehyde Int-8b (1.53 g, 12.26 mmol) was added and the reaction mixture was stirred for 2 hours before being quenched. 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 to give Int-8c (1.5 g, yield 91%) as yellow oil. 1HNMR (500 MHz, DMSO-d6) δ 10.04 (s, 1H), 8.79-8.70 (m, 2H), 8.37 (dd, J = 8.6, 2.3 Hz, 1H), 8.05 (d, J = 1.0 Hz, 2H), 7.42 (d, J = 8.6 Hz, 1H).

[0207] Second Step: Compound Int-8c (1.53 g, 5.70 mmol) was dissolved in ethanol (5 mL), hydroxylamine hydrochloride (792 mg, 11.41 mmol) was added, and the mixture was stirred at room temperature overnight. The reaction mixture was concentrated to give the crude oxime, which was dissolved in acetic acid (5 mL), and zinc powder (1.94 g, 29.66 mmol) was added. The mixture was stirred at room temperature for 2 hours, and LCMS indicated that the reaction was complete. The reaction mixture was filtered, most of the acetic acid was removed by concentration, and the mixture was diluted with ethyl acetate and then basified to pH = 11 with NaOH solution (2 N). The mixture was filtered, and the filtrate was concentrated to give compound Int-8 (1.3 g), which was used directly in the next step. ESI-MS (m / z): 270.5 [M+H] + .

[0208] Intermediate 9

[0209]

[0210] Intermediate 9 was prepared by the following steps:

[0211]

[0212] First Step: Compound Int-9a (500 mg, 3.83 mmol), 6-fluoro-nicotinonitrile Int-5b (514.47 mg, 4.21 mmol), and cesium carbonate (1.87 g, 5.75 mmol) were dissolved in acetonitrile (10 mL), and the mixture was stirred at room temperature overnight. LCMS indicated that the reaction was complete. The reaction mixture was diluted with ethyl acetate, washed with water and saturated brine successively, and the organic phase was dried over anhydrous sodium sulfate. After concentration, the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1) to give Int-9b (630 mg, yield 70%) as a yellow oil. ESI-MS (m / z): 233.4 [M+H] + .

[0213] Second Step: Dissolve and disperse compound Int-9b (630 mg, 2.71 mmol) and Raney Nickel (0.5 mL, aqueous suspension) in methanol (10 mL) and ammonia (1 mL), replace the gas with a hydrogen balloon, and stir at room temperature overnight. Monitor the reaction by LCMS until completion. Dilute the reaction mixture with methanol, filter the reaction mixture through a pad of celite, dry the organic phase over anhydrous sodium sulfate, and concentrate to obtain Int-9 (200 mg, 31% yield) as a yellow oil. ESI-MS (m / z): 237.4 [M+H] + .

[0214] Intermediate 10

[0215]

[0216] Intermediate 10 is prepared by the following steps:

[0217]

[0218] First Step: Dissolve 2-(methylthio)-5-hydroxyimidazole Int-10a (300 mg, 2.11 mmol) in acetonitrile (5 mL), add cesium carbonate (1.37 g, 4.22 mmol), stir at room temperature for 30 minutes, then add 2-fluoropyridine-5-carbaldehyde Int-8b (316 mg, 2.53 mmol), and stir the reaction mixture overnight. Monitor the reaction by LCMS until completion. Dilute the reaction mixture with water, extract with ethyl acetate, dry the organic phase over anhydrous sodium sulfate, filter, and concentrate to obtain Int-10b (511 mg, 97% yield) as a yellow solid. ESI-MS (m / z): 248.5 [M+H] + .

[0219] Second Step: Dissolve compound Int-10b (400 mg, 1.62 mmol) and 3,4-dimethoxybenzylamine (405 mg, 2.43 mmol) in dichloromethane (5 mL) and methanol (0.5 mL), stir at room temperature for 2 hours, then add sodium borohydride in acetic acid (2.06 g, 9.71 mmol), and stir the reaction mixture overnight. Monitor the reaction by LCMS until completion. Dilute the reaction mixture with water, extract with ethyl acetate, dry the organic phase over anhydrous sodium sulfate, filter, and concentrate to obtain Int-10c (630 mg, 98% yield) as a yellow liquid. ESI-MS (m / z): 399.4 [M+H] + .

[0220] Step 3: Compound Int-10c (630 mg, 1.58 mmol), di-tert-butyl dicarbonate (345 mg, 1.58 mmol) and triethylamine (239 mg, 2.37 mmol) were dissolved in dichloromethane (5 mL), after stirring at room temperature for 2 hours, the reaction was monitored to end by LCMS. The reaction was diluted with water, extracted with ethyl acetate, 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 Int-10d (402 mg, yield 51%) as a yellow oil. ESI-MS (m / z): 499.4 [M+H] + .

[0221] Step 4: Compound Int-10d (402 mg, 0.86 mmol) was added to trifluoroacetic acid (1.5 mL), stirred at room temperature overnight, the reaction was monitored to end by LCMS. The reaction was diluted with water, the solution was adjusted to PH = 8 with sodium hydroxide, extracted with dichloromethane, 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 Int-10 (80 mg, yield 39%) as a yellow oil. ESI-MS (m / z): 249.4 [M+H] + .

[0222] The synthesis method of the compound of the example in the application is as follows:

[0223] Example 1

[0224] (S)-4,7,8-trimethyl-2-(((6-((2-(trifluoromethyl)pyrimidin-5-yl)oxy)pyridin-3-yl)methyl)amino)-7,8-dihydropteridine

[0225] -6(5H)-ketone

[0226]

[0227] Example 1 is prepared from the following steps:

[0228]

[0229] Step 1: Compound Int-1 (50 mg, 220.59 umol) and compound Int-7 (77.49 mg, 286.77 umol) were dissolved in n-butanol (2 mL), p-toluenesulfonic acid monohydrate (3.89 mg, 27 umol) was added, the reaction was stirred at 160°C under microwave conditions for 3 hours. After the reaction was cooled to room temperature, the reaction was concentrated under reduced pressure, and the residue was purified by Pre-HPLC to obtain white solid 1 (21.43 mg, yield 21%). ESI-MS (m / z): 461.2 [M+H]+ ; 1 H NMR (500 MHz, DMSO-d6) δ 10.35 (s, 1H), 9.06 (s, 2H), 8.20 (d, J = 2.3 Hz, 1H), 7.99 (dd, J = 8.5, 2.4 Hz, 2H), 7.29 (d, J = 8.4 Hz, 1H), 4.55 (d, J = 6.0 Hz, 2H), 4.23 (d, J = 6.9 Hz, 1H), 3.07 (s, 3H), 2.24 (s, 3H), 1.36 (d, J = 6.9 Hz, 3H).

[0230] Example 2

[0231]

[0232] (S)-4,7-Dimethyl-8-(methyl-d3)-2-(((6-((2-(trifluoromethyl)pyrimidin-5-yl)oxy)pyridin-3- yl)methyl)amino)-7,8-dihydropteridin-6(5H)-one

[0233] Example 2 was prepared from the following steps:

[0234]

[0235] First Step: Compound Int-2 (100 mg, 435.39 umol) and compound Int-7 (152.94 mg, 566.00 umol) were dissolved in n-butanol (2 mL), p-toluenesulfonic acid monohydrate (8.27 mg, 43.54 umol) was added, the reaction was stirred at 160 °C under microwave condition for 3 hours. The reaction was cooled to room temperature, the reaction was concentrated under reduced pressure, the residue was purified by Prep-HPLC to give white solid 2 (38.22 mg, yield 18%). ESI-MS (m / z): 464.2 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) δ 10.35 (s, 1H), 9.06 (s, 2H), 8.20 (d, J = 2.3 Hz, 1H), 7.99 (dd, J = 8.5, 2.4 Hz, 2H), 7.29 (d, J = 8.4 Hz, 1H), 4.55 (d, J = 6.0 Hz, 2H), 4.23 (d, J = 6.9 Hz, 1H), 3.07 (s, 3H), 2.24 (s, 3H), 1.36 (d, J = 6.9 Hz, 3H).

[0236] Example 3

[0237] (S)-4-methyl-2-(((6-((2-(trifluoromethyl)pyrimidin-5-yl)oxy)pyridin-3-yl)methyl)amino)- 6a,7,8,9-tetrahydropyrrolo[2,1-h]pteridin-6(5H)-one

[0238]

[0239] Example 3 was prepared from the following steps:

[0240]

[0241] First Step: Compound Int-6 (80 mg, 335.19 umol) and compound Int-7 (117.74 mg, 435.74 umol) were dissolved in n-butanol (2 mL), p-toluenesulfonic acid monohydrate (6.37 mg, 33.52 umol) was added, the reaction was stirred at 160 °C under microwave condition for 3 hours. The reaction was cooled to room temperature, the reaction was concentrated under reduced pressure, the residue was purified by Pre-HPLC to give white solid 3 (99.37 mg, yield 62%). ESI-MS (m / z): 473.2 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) d 9.74 (s, 1H), 9.03 (d, J = 1.9 Hz, 2H), 8.12 (d, J = 2.4 Hz, 1H), 7.92 (dd, J = 8.5, 2.4 Hz, 1H), 7.21 (d, J = 8.4 Hz, 1H), 6.98 (t, J = 6.4 Hz, 1H), 4.46 - 4.26 (m, 2H), 4.02 - 3.92 (m, 1H), 3.60 - 3.48 (m, 1H), 3.44 - 3.35 (m, 1H), 2.19 - 2.14 (m, 1H), 2.10 (s, 3H), 1.99 - 1.79 (m, 3H).

[0242] Example 4

[0243] 4',8'-dimethyl-2'-(((6-((2-(trifluoromethyl)pyrimidin-5-yl)oxy)pyridin-3-yl)methyl)amino)- 5',8'-dihydro-6'H-spiro[cyclopropane-1,7'-pteridine]-6'-one

[0244]

[0245] Example 4 was prepared from the following steps:

[0246]

[0247] First Step: Sodium hydride (1.59 g, 39.76 mmol, 60% content) was added to a two-necked flask containing anhydrous tetrahydrofuran (20 mL), which was placed in an ice water bath. Compound Int-4a (2 g, 9.94 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL) and slowly added to the reaction solution. After 30 minutes, iodomethane (3.53 g, 24.85 mmol) was slowly added. After the addition was complete, the reaction was slowly warmed to room temperature and stirred overnight. The reaction was monitored to completion by LCMS. The reaction was quenched by slowly adding saturated ammonium chloride aqueous solution at 0 °C. The reaction was extracted with ethyl acetate, and the organic phase was dried to obtain yellow oil 4a (1.7 g, yield 79%). ESI-MS (m / z): 216.2 [M+H] + .

[0248] Second Step: Compound 4a (1.7 g, 7.90 mmol) was dissolved in methanol (5 mL) and placed in an ice water bath. Sulfoxyl chloride (2.82 g, 23.69 mmol) was slowly added. After the addition was complete, the reaction was warmed to room temperature and then to 70 °C and stirred overnight. After the reaction was monitored to completion by LCMS, the reaction was concentrated to obtain yellow solid 4b (1.2 g, yield 91%). ESI-MS (m / z): 130.2 [M+H] + .

[0249] Third Step: Compound Int-3d (500 mg, 2.40 mmol) and compound 4b (597.17 mg, 3.61 mmol) were dissolved in tetrahydrofuran (10 mL), and N,N-diisopropylethylamine (1.26 mL, 7.20 mmol) was added. The reaction was stirred at room temperature for 8 hours. The reaction was monitored to completion by LCMS. The reaction was diluted with water and extracted with ethyl acetate. The reaction was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1) to obtain yellow oil 4c (200 mg, reaction yield 27%). ESI-MS (m / z): 301.3 [M+H] + .

[0250] Fourth Step: Compound 4c (200 mg, 665.12 umol) and compound Int-7 (270.21 mg, 215.67 umol) were dissolved in N,N-dimethylformamide (5 mL), and N,N-diisopropylethylamine (27.89 mg, 2.00 mmol) was added. The reaction was stirred at room temperature for 8 hours. The reaction was monitored to completion by LCMS. The reaction was diluted with water and extracted with ethyl acetate. The reaction was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to obtain compound 4d (300 mg, yield 84%). ESI-MS (m / z): 535.2 [M+H] + .

[0251] Step 5: Compound 4d (250 mg, 467.77 umol) was dissolved in methanol (5 mL), palladium on carbon (5.68 mg, 46.78 umol) was added, and then the reaction system was replaced with hydrogen. The reaction was stirred at room temperature for 8 hours. LCMS monitored the end of the reaction, the residue was filtered with diatomite, and the reaction solution was concentrated under reduced pressure. The residue was purified by Prep-HPLC to obtain white solid 4 (24.02 mg, yield 10%). ESI-MS (m / z): 473.2 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) δ 9.87 (s, 1H), 9.03 (s, 2H), 8.11 (d, J = 2.3 Hz, 1H), 7.91 (dd, J = 8.4, 2.2 Hz, 1H), 7.22 (d, J = 8.4 Hz, 1H), 7.10-6.90 (m, 1H), 4.36 (d, J = 6.4 Hz, 2H), 2.72 (s, 3H), 2.11 (s, 3H), 1.37-1.20 (m, 2H), 1.20-1.07 (m, 2H).

[0252] Example 5

[0253] 4'-methyl-8'-(methyl-d3)-2'-(((6-((2-(trifluoromethyl)pyrimidin-5-yl)oxy)pyridin-3-yl)methyl)amino)-5',8'-dihydro

[0254] -6'H-spiro[cyclopropane-1,7'-pteridine]-6'-one

[0255]

[0256] Example 5 was obtained by referring to the synthetic method of Example 4. ESI-MS (m / z): 476.7 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) δ 9.87 (s, 1H), 9.03 (s, 2H), 8.11 (d, J = 2.3 Hz, 1H), 7.91 (dd, J = 8.4, 2.2 Hz, 1H), 7.22 (d, J = 8.4 Hz, 1H), 7.10-6.90 (m, 1H), 4.36 (d, J = 6.4 Hz, 2H), 2.72 (s, 3H), 2.11 (s, 3H), 1.37-1.20 (m, 2H), 1.20-1.07 (m, 2H).

[0257] Example 6

[0258] 4,7,7-Trimethyl-8-(methyl-d3)-2-(((6-((2-(trifluoromethyl)pyrimidin-5-yl)oxy)pyridin-3- yl)methyl)amino)-7,8-dihydropteridin-6(5H)-one

[0259]

[0260] Example 6 was prepared from the following steps:

[0261]

[0262] First Step: Compound Int-3 (55 mg, 225 umol) and compound Int-7 (73 mg, 270 umol) were dissolved in n-butanol (2 mL), p-toluenesulfonic acid monohydrate (3.89 mg, 27 umol) was added, the reaction was stirred at 160 °C under microwave condition for 3 hours. The reaction was cooled to room temperature, the reaction was concentrated under reduced pressure, the residue was purified by Pre-HPLC to give white solid 6 (24 mg, yield 21%). ESI-MS (m / z): 478.6 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) d 9.80 (s, 1H), 9.04 (s, 2H), 8.13 (d, J = 2.3 Hz, 1H), 7.93 (dd, J = 8.4, 2.4 Hz, 1H), 7.23 (d, J = 8.4 Hz, 1H), 6.99 (t, J = 6.2 Hz, 1H), 4.38 (d, J = 6.3 Hz, 2H), 2.13 (s, 3H), 1.32 (s, 6H).

[0263] Example 7

[0264] 4,7,7,8-Tetramethyl-2-(((6-((2-(trifluoromethyl)pyrimidin-5-yl)oxy)pyridin-3-yl)methyl)amino)- 7,8-dihydropteridin-6(5H)-one

[0265]

[0266] Example 7 was prepared from the following steps:

[0267]

[0268] First Step: Sodium hydride (275 mg, 6.89 mmol, 60% content) was added to a two-necked flask containing anhydrous tetrahydrofuran (5 mL) and placed in an ice-water bath. Compound 7a (350 mg, 1.72 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL) and slowly added to the reaction solution. After 30 minutes, iodomethane (366 mg, 2.58 mmol) was added dropwise, and after the addition was complete, the reaction was slowly warmed to room temperature and stirred overnight. The reaction was monitored to completion by LCMS. The reaction was quenched by slowly adding saturated aqueous ammonium chloride solution at 0 °C, and extracted with ethyl acetate. The organic phase was dried to obtain yellow oil 7b (370 mg, yield 98%). ESI-MS (m / z): 218.7 [M+H] + .

[0269] Second Step: Compound 7b (370 mg, 1.70 mmol) was dissolved in methanol (5 mL) and placed in an ice-water bath. Sulfoxide chloride (405 mg, 3.41 mmol) was slowly added dropwise, and after the addition was complete, the reaction was warmed to room temperature and then to 70 °C and stirred overnight. After the reaction was monitored to completion by LCMS, the reaction was concentrated to obtain yellow oil 7c (200 mg, yield 86%). ESI-MS (m / z): 132.7 [M+H] + .

[0270] Third Step: Compound 7c (200 mg, 1.19 mmol) and compound Int-3d (225 mg, 1.08 mmol) were dissolved in tetrahydrofuran (5 mL), and N,N-diisopropylpropylamine (420 mg, 3.25 mmol) was added. The reaction was stirred at room temperature for 8 hours. The reaction was monitored to completion by LCMS. The reaction was diluted with water and extracted with ethyl acetate. The reaction was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1) to obtain white solid 7d (70 mg, reaction yield 21%). ESI-MS (m / z): 303.3 [M+H] + .

[0271] Fourth Step: Compound 7d (70 mg, 231 umol) and compound Int-7 (75 mg, 278 umol) were dissolved in tetrahydrofuran (5 mL), and N,N-diisopropylpropylamine (90 mg, 694 umol) was added. The reaction was stirred at room temperature for 8 hours. The reaction was monitored to completion by LCMS. The reaction was diluted with water and extracted with ethyl acetate. The reaction was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to obtain compound 7e (90 mg, yield 72%). ESI-MS (m / z): 538.7 [M+H] + .

[0272] Fifth Step: Compound 7e (90 mg, 177 umol) was dissolved in methanol (5 mL), palladium on carbon (10 mg, 89 umol) was added, and the reaction system was replaced with hydrogen gas. The reaction was stirred at room temperature for 8 hours. LCMS monitored the end of the reaction, the residue was filtered with diatomite, and the reaction was concentrated under reduced pressure. The residue was purified by Prep-HPLC to obtain white solid 7 (21 mg, yield 25%). ESI-MS (m / z): 475.7 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) δ 9.82 (s, 1H), 9.04 (s, 2H), 8.13 (d, J = 2.3 Hz, 1H), 7.93 (dd, J = 8.4, 2.4 Hz, 1H), 7.23 (d, J = 8.4 Hz, 1H), 7.00 (t, J = 6.3 Hz, 1H), 4.38 (d, J = 6.3 Hz, 2H), 2.94 (s, 3H), 2.14 (s, 3H), 1.33 (s, 6H).

[0273] Example 8

[0274] (S)-2-(((6-((2-chloropyrimidin-5-yl)oxy)pyridin-3-yl)methyl)amino)-4,7,8- trimethyl-7,8-dihydropteridin-6(5H)-one

[0275]

[0276] Example 8 was prepared from the following steps:

[0277]

[0278] First Step: Compound Int-1 (50.00 mg, 220.59 umol) and compound Int-9 (67.87 mg, 286.77 umol) were dissolved in n-butanol (2 mL), p-toluenesulfonic acid monohydrate (4.19 mg, 22.06 umol) was added, and the reaction was stirred at 160 °C under microwave conditions for 3 hours. After the reaction was cooled to room temperature, the reaction was concentrated under reduced pressure, and the residue was purified by Prep-HPLC to obtain white solid 8 (10.69 mg, yield 11%). ESI-MS (m / z): 427.6 [M+H] + ; 1H NMR(500MHz,DMSO-d6)δ9.82(s,1H),8.78(s,2H),8.10(d,J=2.3Hz,1H),7.89(dd,J=8.5,2.4Hz,1H),7.17(d,J=8.5Hz ,1H),6.98(t,J=6.3Hz,1H),4.42-4.30(m,2H),3.99(q,J=6.8Hz,1H),2.91(s,3H),2.11(s,3H),1.18(d,J=6.8Hz,3H).

[0279] Example 9

[0280] (S)-4,7,8-Trimethyl-2-(((6-((6-(trifluoromethyl)pyridin-3-yl)oxo)pyridin-3-yl)methyl)amino)-7,8-dihydropteridin-6(5H)-one

[0281]

[0282] Example 9 was prepared by the following steps:

[0283]

[0284] Step 1: Compound Int-1 (50.00 mg, 220.59 μmol) and compound Int-8 (77.02 mg, 286.77 μmol) were dissolved in n-butanol (2 mL), and p-toluenesulfonic acid monohydrate (4.19 mg, 22.06 μmol) was added. The reaction mixture was stirred at 160 °C for 3 hours under microwave conditions. After the reaction mixture cooled to room temperature, it was concentrated under reduced pressure. The residue was purified by Pre-HPLC to give a white solid 9 (64.26 mg, yield 63%). ESI-MS (m / z): 460.6 [M+H] + ; 1 H NMR (500MHz, DMSO-d6) δ9.80(s,1H),8.62(s,1H),8.12(s,1H),7.95(d,J=8.6Hz,1H),7.89(d,J=8.4Hz,1H),7.85(d,J=8.6Hz,1H),7. 15(d,J=8.5Hz,1H),6.95(t,J=6.3Hz,1H),4.46-4.28(m,2H),3.98(q,J=6.9Hz,1H),2.91(s,3H),2.11(s,3H),1.17(d,J=6.8Hz,3H).

[0285] Example 10

[0286] (S)-2-(((6-((6-chloropyridin-3-yl)oxy)pyridin-3-yl)methyl)amino)-4,7,8- trimethyl-7,8-dihydropteridin-6(5H)-one

[0287]

[0288] Example 10 was prepared from the following steps:

[0289]

[0290] First Step: Compound Int-1 (50.00 mg, 220.59 umol) and compound Int-5 (67.58 mg, 286.77 umol) were dissolved in n-butanol (2 mL), p-toluenesulfonic acid monohydrate (4.19 mg, 22.06 umol) was added, the reaction was stirred at 160 °C under microwave condition for 3 hours. The reaction was cooled to room temperature, the reaction was concentrated under reduced pressure, the residue was purified by Pre-HPLC to give white solid 10 (49.62 mg, yield 52%). ESI-MS (m / z): 426.6 [M+H] + ; 1 H NMR (500 MHz, Chloroform-d) δ 8.19 (d, J = 2.9 Hz, 1H), 8.06 (d, J = 2.4 Hz, 1H), 7.70 (dd, J = 8.4, 2.5 Hz, 1H), 7.42 (dd, J = 8.6, 2.9 Hz, 1H), 7.27 (d, J = 8.6 Hz, 1H), 6.88 (d, J = 8.4 Hz, 1H), 5.34 (s, 1H), 4.54 - 4.39 (m, 2H), 4.00 (q, J = 6.9 Hz, 1H), 2.95 (s, 3H), 2.15 (s, 3H), 1.33 (d, J = 6.9 Hz, 3H).

[0291] According to the above described synthetic routes and methods of synthesizing intermediates, the following examples can be obtained.

[0292]

[0293]

[0294] Comparative Example 1

[0295] (S)-2-(((6-(4-fluorophenoxy)pyridin-3-yl)methyl)amino)-4,7,8-trimethyl-7,8-dihydropteridin-6

[0296] (5H)-one

[0297]

[0298] Example 1 was obtained by referring to the synthetic method of compound 54 described in patent WO2019209757. ESI-MS (m / z): 409.8 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) δ 9.80 (s, 1H), 8.07 (d, J = 2.3 Hz, 1H), 7.79 (dd, J = 8.6, 2.3 Hz, 1H), 7.28 - 7.18 (m, 2H), 7.18 - 7.08 (m, 2H), 7.01 - 6.88 (m, 2H), 4.44 - 4.26 (m, 2H), 3.98 (q, J = 6.9 Hz, 1H), 2.91 (s, 3H), 2.11 (s, 3H), 1.17 (d, J = 6.7 Hz, 3H).

[0299] Comparative Example 2

[0300] (S)-2-(((6-(4-fluorophenoxy)pyridin-3-yl)methyl)amino)-7,8-dimethyl-7,8-dihydropteridin-6(5

[0301] H)-one

[0302]

[0303] Example 2 was obtained by referring to the synthetic method of compound 56 described in patent WO2019209757. ESI-MS (m / z): 395.7 [M+H] + ; 1 H NMR (500 MHz, Chloroform-d) δ 8.11 (d, J = 2.3 Hz, 1H), 7.66 (dd, J = 8.5, 2.4 Hz, 1H), 7.19 (s, 1H), 7.03 - 6.96 (m, 4H), 6.77 (d, J = 8.4 Hz, 1H), 4.54 - 4.38 (m, 2H), 4.01 (q, J = 6.9 Hz, 1H), 2.97 (s, 3H), 1.38 (dd, J = 6.9, 1.3 Hz, 3H).

[0304] Biological screening of Wnt pathway inhibitors and results

[0305] Test Example 1: Construction of Colo205-LUC-TCF / LEF-M1 reporter cell line

[0306] Colo205 cell line (Chinese Academy of Sciences Cell Bank, Cat#TCHu102) was purchased from Chinese Academy of Sciences Cell Bank, after expansion and subculture, the cells were in the exponential growth phase, and then transfected with lipo3000 liposome to transfect the luciferase reporter plasmid (Promega) driven by TCF / LEF transcription factor. The plasmid contains a resistance gene, which can be used for resistance screening. Transfection was performed in a 10 cm culture dish using regular complete medium without antibiotics. 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 medium was centrifuged to remove cells and debris, and then 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, with an average of 1.5 cells 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 were labeled as single clone wells. Then the culture was continued, the medium was replaced every 2 days, and the observation was continued. 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 cells in the 96-well plate 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 method. 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.

[0307] Test Example 2: Detection of the inhibitory ability of the compound on Colo205-LUC-TCF / LEF M1 reporter cell line

[0308] Colo205-LUC-TCF / LEF M1 cell line is a stable transfection of pGL4.49-LUC2-TCF / LEF vector reporter tool cell, its β-catenin Wnt pathway is continuously activated, after adding inhibitor, Wnt pathway is inhibited, TCF / LEF cis element regulated firefly luciferase expression on the vector decreases, after adding detection substrate, the detected light signal decreases accordingly, thereby detecting the inhibitory effect of the compound.

[0309] To each well of a 96-well cell culture plate, 100 μL of a compound was added at a maximum concentration of 20 μM, and the compound concentration was diluted by 3-fold gradient. Then, 10000 stable-transfected reporter gene colo205 cells and 100 μL of culture medium were inoculated into each well, and the corresponding treatment was performed as positive and negative control wells. The cells were placed in a 5% CO2 cell incubator at 37°C for 4 h, and after 4 h, the culture medium was removed, and 100 μL of a reagent (Promega) containing the corresponding luciferase substrate was added to each well to determine the luciferase reporter gene activity. The luminescence intensity was read by SpectraMax in full wavelength mode. The light signal intensity of cells treated only by DMSO was the positive control, and the light signal intensity of the wells without cells was the negative control, and the IC 50 values of each compound were calculated. The colo 205 reporter gene detection data are summarized in Table 1 below.

[0310] Table 1 IC values of compounds for inhibition of Colo205-LUC-TCF / LEF reporter gene 50

[0311]

[0312]

[0313] Test Example 3: Proliferation inhibition test of compounds on Wnt mutant cell lines (Colo205, DU4475, NCI-H929 and HepG2) and non-Wnt mutant cell lines (Hela and RKO)

[0314] The cell lines used in the test are Colo205, DU4475, NCI-H929 and HepG2 cell lines whose Wnt pathway is continuously activated and whose proliferation is dependent on the Wnt pathway, and HE LA and RKO cell lines which are not activated under normal circumstances and whose proliferation is not dependent on the Wnt pathway as control cell lines, so as to determine whether the inhibition of Wnt-dependent proliferation of the compounds of the present application is caused by other non-specific toxicity.

[0315] ​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 and cultured 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 the culture was continued 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 cell inoculation was used as the negative control well, and the IC 50 values of each compound for the proliferation inhibition of Wnt continuously activated or proliferation dependent cells, and the IC 50 values of each compound for the proliferation inhibition of Wnt non-activated or proliferation independent cells were calculated to evaluate the inhibitory effect of the compound on the Wnt pathway and the toxic effect on normal cells. The results are shown in Table 2 below.

[0316] Table 2 IC 50 values of compounds for the proliferation inhibition of Wnt mutant cell lines

[0317]

[0318]

[0319] NT represents not detected

[0320] The above results show that the compounds of the present application have significant inhibitory activity on mutant cell lines Colo205, DU4475, NCI-H929 and HepG2, and have substantially no significant inhibitory activity on Hela and RKO cell lines, which indicates that the compounds of the present application have significant Wnt-dependent proliferation inhibition effect.

[0321] Test Example 4: Mouse pharmacokinetic evaluation of the compound of the present application

[0322] 1. Experimental materials

[0323] ICR mice: male, 25-35g, purchased from Shanghai Slek Experimental Animal Co., Ltd.

[0324] Reagents: DMSO (dimethyl sulfoxide), PEG-400 (polyethylene glycol 400), Solutol HS-15, ultrapure water, methanol, acetonitrile, formic acid, propranolol (internal standard), tolbutamide (internal standard), ICR mouse blank plasma (EDTA-K2 anticoagulant)

[0325] Instrument: SCIEX LC-MS / MS (Exion LC, QTRAP 6500 Plus)

[0326] 2. Experimental method

[0327] A certain amount of compounds of Examples 1, 2, 3, 5, 6, 9, and 10 and the compound of Comparative Example 1 was weighed, dissolved in 5% DMSO, and then other corresponding solvents (see Table 3) were added to prepare a clear solution. After intravenous or oral administration to mice, 100 μL of blood was collected from the orbit at 15 min, 30 min, 1 h, 2 h, 4 h, 8 h, and 24 h (5 min was added for the intravenous administration group), and placed in an EDTA-K2 anticoagulant centrifuge tube. The blood plasma was separated by centrifugation (4°C) at 3000g for 10 min within 30 min, and stored at -80°C for testing.

[0328] A certain amount of compound was accurately weighed and dissolved in DMSO to prepare a stock solution at 2 mg / ml. An appropriate amount of the compound stock solution was accurately pipetted, diluted with an acetonitrile-water (8:2) solution to prepare a standard series solution, and 1 μL of the standard series solution was accurately pipetted and mixed with 19 μL of mouse blank plasma to prepare a plasma matrix standard curve with a series of concentrations. 20 μL of each mouse plasma sample after administration was accurately pipetted, 400 μL of an internal standard-containing acetonitrile solution (propranolol 50 ng / ml, tolbutamide 100 ng / ml) was added, vortexed for 5 min, centrifuged at 4000 rpm for 10 min, and 200 μL of supernatant was taken and mixed with 200 μL of ultrapure water. The mixture was then analyzed by LC-MS / MS.

[0329] 3. Data processing

[0330] After the blood drug concentration was determined by LC-MS, the pharmacokinetic parameters were calculated by WinNonlin 8.1 software using a non-compartment model, and the results are shown in Table 3.

[0331] Table 3 Pharmacokinetic parameters of compounds in mice

[0332]

[0333] Solvent A: 5% DMSO + 10% PEG-400 + 10% Solutol + 75% H2O

[0334] B: 5% DMSO + 10% PEG 400 + 85% H2O

[0335] C: 5% DMSO + 20% Solutol + 75% H2O

[0336] The above results show that the highest blood drug concentration (Cmax) and the area under the absorption curve (AUC) detected after oral administration of the compound of the present application to mice are significantly better than the control. This shows that the new structural feature has the ability to improve and enhance the oral absorption characteristics.

[0337] Test Example 5: Tumor growth inhibition test of compound 1 (W421) on Colo205 mouse xenograft model

[0338] The in vivo anti-tumor activity of compound 1 was evaluated in a BALB / c Nude mouse xenograft model of human colon cancer cell Colo205.

[0339] Female BALB / c Nude mice were subcutaneously inoculated with human colon cancer cell Colo205 to establish a Colo205 BALB / c Nude mouse xenograft model. When the tumors grew to an average tumor volume of 100 mm 3 After 7 days, the tumor-bearing mice were randomly divided into 4 groups according to the tumor size: solvent treatment control group, 1.5 mg / kg compound 1 group, 3 mg / kg compound 1 group and 10 mg / kg compound 1 group. Compound 1 was administered orally, once a day, for 15 days, and the tumor volume was measured every other day (Table 4). Figure 1 On Day 15, the body weight and tumor volume were measured (Table 4).

[0340] Table 4 Inhibition of tumor volume of human colon cancer cell Colo205 by compound 1 in vivo (day 15)

[0341]

[0342] a. Mean ± SEM.

[0343] b. Tumor growth inhibition was calculated from T / C (TRTV / CRTV) (TRTV: average RTV of the treatment group; CRTV: average RTV of the vehicle control group; RTV = Vt / V0, V0 is the tumor volume of the animal at the time of grouping, and Vt is the tumor volume of the animal after treatment).

[0344] c. p value was calculated according to the tumor volume, and one-way ANOVA was used for analysis between groups. If the variance homogeneity test p > 0.05, Dunnett t was selected, otherwise Games-Howell was selected.

[0345] Figure 1 The results of Table 4 show that the compound of the present application, in particular compound 1, has a significant function of inhibiting the tumor volume of human colon cancer cell Colo205 in vivo.

Claims

1. A compound or a pharmaceutically acceptable salt, isotope derivative, or stereoisomer thereof, wherein the compound has the following structure:

2. A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt, isotope derivative, stereoisomer thereof, and optionally a pharmaceutically acceptable carrier.

3. Use of the compound of claim 1 or a pharmaceutically acceptable salt, isotope derivative, stereoisomer, or pharmaceutical composition of claim 2 in the preparation of a medicament for the prevention and / or treatment of tumors, inflammatory diseases, or immune-mediated diseases.

4. Use of the compound of claim 1 or a pharmaceutically acceptable salt, isotope derivative, stereoisomer, or pharmaceutical composition of claim 2 in the preparation of a medicament for the prevention and / or treatment of cancer or autoimmune diseases.

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