CN103387535A - Substituted alkynylpyridine compounds and methods and uses thereof - Google Patents

Abstract

The present invention provides a substituted pyridine compound having a structure of formula (I), a pharmaceutically acceptable salt thereof, and a pharmaceutical preparation thereof, which is used to regulate the activity of protein kinases and regulate intercellular or intracellular signal responses. The present invention also relates to a pharmaceutical composition comprising the compound of the present invention, and a method for using the pharmaceutical composition to treat mammals, especially human hyperproliferative diseases.

Description

Substituted alkynylpyridine compounds and methods and uses thereof

This application requires a Chinese patent application submitted to the China Patent Office on May 10, 2012, with the application number 201210141993.5, and the title of the invention is "Substituted Alkynylpyridine Compounds and Their Use and Application" and submitted on April 3, 2013 China Patent Office, the priority of the Chinese patent application with the application number 201310116652.7 and the title of the invention "Substituted alkynylpyridine compounds and their use methods and uses", the entire content of which is incorporated in this application by reference.

technical field

The invention belongs to the technical field of medicines, and in particular relates to a substituted alkynylpyridine compound and its application method and application.

Background technique

Protein kinase, as an important regulator of cell function, is one of the members with the largest number and widest functions in the gene family. By adding phosphate groups to substrate proteins, they regulate the activity, location and overall function of a variety of proteins and are involved in orchestrating many cellular processes. Kinases play a prominent role in signal transduction and the coordination of complex functions such as the cell cycle. Among the 518 human protein kinases, 478 were classified into a superfamily due to their similar catalytic domain sequences, and they could be divided into different groups, families or subfamilies according to the similarity of the growing sequence and biochemical activity.

Wherein the partial list of kinases includes abl, AATK, ALK, Akt, axl, bmx, bcr-abl, Blk, Brk, Btk, csk, c-kit, c-Met, c-src, c-fins, CDK1, CDK2 , CDK3, CDK4, CDK5, CDK6, CDK7, CDK8, CDK9, CDK10, cRaf1, CSF1R, CSK, DDR1, DDR2, EPHA, EPHB, EGFR, ErbB2, ErbB3, ErbB4, Erk, Fak, fes, FER, FGFR1, FGFR2 , FGFR3, FGFR4, FGFR5, Fgr, flt-1, Fps, Frk, Fyn, GSG2, GSK, Hck, ILK, INSRR, IRAK4, ITK, IGF-1R, INS-R, Jak, KSR1, KDR, LMTK2, LMTK3 , LTK, Lck, Lyn, MATK, MERTK, MLTK, MST1R, MUSK, NPR1, NTRK, MEK, PLK4, PTK, p38, PDGFR, PIK, PKC, PYK2, RET, ROR1, ROR2, RYK, ros, Ron, SGK493 , SRC, SRMS, STYK1, SYK, TEC, TEK, TEX14, TNK1, TNK2, TNNI3K, TXK, TYK2, TYRO3, tie, tie2, TRK, Yes, and Zap70.

Receptor tyrosine kinases (RTKs) are an abundant class of transmembrane proteins that serve as receptors for cytokines, growth factors, hormones, and other signaling molecules. Receptor tyrosine kinases are expressed in many types of cells and play important roles in a variety of cellular processes, including cell growth, differentiation, and angiogenesis. Kinase activation begins with ligand binding in the extracellular domain, which in turn causes a conformational change that leads to receptor dimerization, reciprocal phosphorylation of dimerized receptors, and subsequent autophosphorylation of tyrosine residues outside the catalytic domain. This autophosphorylation both stabilizes the conformation of activated receptors and creates phosphorylation stacking sites in intracellular signaling proteins.

Receptor tyrosine kinases are highly active in many human solid tumors and hematological malignancies through receptor activating mutations, gene amplification, and growth factor activation. Accelerated activation of RTKs contributes to various tumorigenic factors, such as proliferation, survival, invasion, metastasis, and angiogenesis, and therefore, inhibition of the activity of receptor tyrosine kinases is considered an effective strategy for cancer therapy (Sharma PS; et al. "Receptor tyrosine kinase inhibitors as potent weapons inwar against cancers." Curr Pharm Des. 2009, 15, 758).

The receptor tyrosine kinase anaplastic lymphoma kinase (ALK), which belongs to the insulin receptor superfamily, is involved in the development of a variety of human tumors. In fact, ALK has been initially identified as a constitutively activated and oncogene fusion to nucleophosmin (NPM)-ALK in anaplastic large cell lymphoma, a separate type of non-Hodgkin's lymphoma -Most common (Morris, S.W.; et al. "Fusion of a kinase gene, ALK, to a nuclear proteinene, NPM, in non-Hodgkin's lymphoma." Science 1994, 263, 1281).

In addition, ALK fusion genes have been found in inflammatory myofibroblastic tumors (IMTs), and in a subspecies of squamous cell carcinoma of the esophagus, the TPM4-ALK fusion gene. Studies have shown that there are multiple ALK gene mutations in both familial and sporadic neuroblastoma. This mutation, present in neuroblastoma cells, causes constitutive ALK phosphorylation and functional decline. In contrast, the rapid growth of cell lines can be inhibited by using sRNA and small molecule ALK inhibitors (Palmer, R.H.; et al. "Anaplastic lymphoma kinase: signaling in development and disease." Biochem. J. 2009, 420, 345).

In recent years, it has been confirmed that in non-small cell lung cancer (NSCLC) cells, there are multiple subtypes of fusion genes composed of part of the echinoderm microtubule-associated protein-like 4 (EML4) gene and ALK gene. EML4-ALK fusion gene transcripts were detected in about 3-7% of NSCLC patients. Experiments in vivo and in vitro have confirmed that the EML4-ALK fusion gene protein has oncogenic transformation activity and has an important impact on human NSCLC (Soda, M.; et al "Identification of the transforming EML4-ALK fusion gene in non-small-cell lung cancer .” Nature 2007, 448, 561).

The fusion gene of ALK shows obvious carcinogenesis, and its abnormal tyrosine kinase activity can enhance cell proliferation and survival, lead to rearrangement of cytoskeleton, and change the shape of cells. In the process of oncogenic ALK signal transduction, ALK interacts with downstream molecules and then activates intracellular signaling pathways. Like most normal and oncogenic tyrosine kinases, ALK fusion genes can activate a variety of different pathways, which are closely related to each other. Connected, overlap each other, and finally form a complex signal transduction network. According to literature reports, there are three most relevant and well-researched pathways: Ras-ERK (extracellular signal-regulated kinase) pathway, JAK3 (Janus kinase 3)-STAT3 pathway and PI3K (phosphatidylinositol 3-kinase)-Akt path. Many sites in these three pathways can mediate the activation of ALK. Taken together, the JAK3-STAT3 pathway and the PI3K-Akt pathway play critical roles in cell survival and phenotypic changes (Chiarle, R.; et al. "Theanaplastic lymphoma kinase in the pathogenesis of cancer." Nat. Rev. Cancer 2008 , 8, 11; Barreca, A.; et al. "Anaplastic lymphoma kinase (ALK) in human cancer." J. Mol. Endocrinol. 2011, 47, R11).

Intact, normal ALK receptors are also associated with the development of other malignancies, such as glioblastoma, neuroblastoma, breast cancer, and others. In a survey of a collection of human cancer cell lines, Dirks et al. demonstrated expression of ALK transcripts in nervous system cell lines and most ectodermal solid cancer cell lines, including retinoblastoma, melanoma tumor and breast cancer (Dirks, P.B. "Cancer's source in the peripheral nervous system." Nature Medicine 2008, 14, 373).

c-Met, namely hepatocyte growth factor receptor (HGFR), its main point of action is endothelial cells, and it has been confirmed that it is expressed in endothelial cells, myogenic cells, hematopoietic cells and motor neurons. The natural ligand of c-Met is hepatocyte growth factor (HGF), which is a multifunctional growth factor, namely scatter factor (SF). In fetuses and adults, activation of c-Met promotes certain morphological, eg, invasive growth, leading to rapid cell growth, cell-to-cell division, and cell migration to their surroundings (Peschard P.; Park M. "From Tpr-Met to Met, tumorigenesis and tubes." Oncogene 2007, 26, 1276; Stellrecht CM; Gandhi V. "Met Receptor Tyrosine Kinase as a Therapeutic Anticancer Target." Cancer Letter 2009, 280, 1).

Persistent c-Met stimulation, overexpression or mutation exists in widespread human malignancies, including breast cancer, liver cancer, lung cancer, ovarian cancer, kidney cancer, thyroid cancer, colon cancer, malignant glioma, prostate cancer, etc. c-Met has also been implicated in atherosclerosis and pulmonary fibrosis. The aggressive growth rate of these cancer cells was radically enhanced through tumor-stroma interactions, including the HGF/c-Met pathway. Thus, substantial evidence linking c-Me signaling responsiveness to the rate of progression of some cancerous diseases raises its role in the development of cancer drugs that primarily target c-Met (Migliore C.; Giordano S." Molecular cancer therapy: canour expectation be MET." Eur. J. Cancer 2008, 44, 641; Benedetta Peruzzi; Donald P. Bottaro. "Targeting the c-Met Signaling Pathway in Cancer." Clinical Cancer Research 2006, 12, 3657). Currently, drugs targeting the c-Met signaling pathway are under clinical research (Joseph Paul Eder; et al. "Novel Therapeutic Inhibitors of the c-Met Signaling Pathway in Cancer." Clinical Cancer Research2009, 15, 2207; "Paolo M.; et al. al. Drug development of MET inhibitors: targeting oncogene addiction and experience." Nature Review Drug Discovery 2008, 7, 504).

代号PF-02341066)用于治疗局部晚期或转移性的，存在间变性淋巴瘤激酶（ALK）基因重排的非小细胞肺癌。ALK(EML4-ALK)基因的重排，导致细胞突变，促进了肺癌细胞的恶性表型。因此，抑制突变的激酶ALK对于治疗癌症是有效的。Clinically, many ALK and/or c-Met inhibitors have been used to treat cancer. Crizotinib is a small-molecule ATP-competitive ALK inhibitor that also acts on c-Met receptor tyrosine kinase. On August 26, 2008, the U.S. FDA approved crizotinib (trade name

Code name PF-02341066) is used for the treatment of locally advanced or metastatic non-small cell lung cancer with anaplastic lymphoma kinase (ALK) gene rearrangement. Rearrangement of the ALK (EML4-ALK) gene, resulting in cellular mutations, promotes the malignant phenotype of lung cancer cells. Therefore, inhibition of the mutated kinase ALK would be effective in the treatment of cancer.

Contents of the invention

In view of this, the present invention provides a new substituted alkynylpyridine compound, which has inhibitory effect on protein tyrosine kinase and can be used to prepare medicines for treating cell proliferation diseases.

The compound of the present invention has inhibitory effect on the activity of protein tyrosine kinase. What is more satisfying is that the compound of the present invention can inhibit ALK (including ALK fusion gene, such as: EML4-ALK, NPM-ALK, etc.), or c -Met receptor (hepatocyte growth factor receptor) signaling response. Correspondingly, the present invention also provides some novel inhibitors of protein tyrosine kinase receptor signal response, such as ALK receptor signal response or c-Met receptor signal response.

In particular, the compounds involved in the present invention, and pharmaceutically acceptable compositions thereof, can be effectively used as inhibitors of tyrosine kinase receptors, such as ALK or c-Met.

In one aspect, the present invention relates to a compound represented by formula (I):

Or its stereoisomer, geometric isomer, tautomer, nitrogen oxide, solvate, metabolite, pharmaceutically acceptable salt or its prodrug, wherein, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , X and Z have the meanings described in the present invention.

In some embodiments, each of R ¹ , R ² , R ³ , R ⁴ , R ⁵ and R ⁶ is independently H, D or F;

X is a C _6-10 aryl group or a heteroaryl group of 5-10 atoms containing 1, 2, 3 or 4 heteroatoms independently selected from O, S or N, wherein the C _6-10 aryl group and a heteroaryl group of 5-10 atoms containing 1, 2, 3 or 4 heteroatoms independently selected from O, S or N is optionally substituted with 1, 2, 3 or 4 substituents, said The substituents are independently selected from D, F, Cl, Br, I, -CN, -NO ₂ , N ₃ , -OR ^a , -SR ^a , -NR ^a R ^b , -C(=O)NR ^a R ^b , C _1-6 alkyl, C _1-6 haloalkyl, C _2-6 alkenyl, C _2-6 alkynyl, -(C _1-4 alkylene)-CN, -(C _1-4 Alkyl)-OR ^a , -(C _1-4 alkylene)-NR ^a R ^b , C _6-10 aryl or containing 1, 2, 3 or 4 heteroatoms independently selected from O, S or N A heteroaryl group of 5-10 atoms;

Z is C _1-6 alkyl, C _3-6 cycloalkyl, C _3-6 heterocyclyl, C _5-12 fused bicyclyl, C _5-12 spiro bicyclyl, -(C _1-4 alkylene Base)-(C _{3-6cycloalkyl} ),-(C _1-4alkylene )-(C _{3-6heterocyclyl} ),-(C _1-4alkylene )-(C _5-12 Fused bicyclyl) or -(C _1-4 alkylene)-(C _5-12 spiro bicyclyl), wherein, the C _1-6 alkyl, C _3-6 cycloalkyl, C _3-6 Heterocyclic group, C _5-12 fused bicyclic group, C _5-12 spirobicyclic group, -(C _1-4 alkylene)-(C _3-6 cycloalkyl), -(C _1-4 alkylene Base)-(C _3-6 heterocyclyl), -(C _1-4 alkylene)-(C _5-12 fused bicyclic group) and -(C _1-4 alkylene)-(C _{5- 12} spirobicyclyl), optionally substituted by 1, 2, 3 or 4 substituents independently selected from D, F, Cl, Br, I, -CN, -NO ₂ , N ₃ , C _1-4 alkyl, -OR ^a , -SR ^a , -NR ^a R ^b or -C(=O)NR ^a R ^b , and, C _5-12 fused bicyclyl, C _5-12 spirobicyclic Each ring in -(C _1-4 alkylene)-(C _5-12 fused bicyclic group) and -(C _1-4 alkylene)-(C _5-12 spiro bicyclic group) is independently is carbocyclic or heterocyclic;

Each of R ^a and R ^b is independently H, C _1-6 aliphatic, C _3-6 cycloalkyl, C _3-6 heterocyclyl, C _6-10 aryl, containing 1, 2, 3 or 4 A heteroaryl group of 5-10 atoms independently selected from heteroatoms of O, S or N, -(C _1-4 alkylene)-(C _3-6 cycloalkyl), -(C _1-4 alkylene Alkyl)-(C _3-6 heterocyclyl), -(C _1-4 alkylene)-(C _6-10 aryl) or -(C _1-4 alkylene)-(including 1,2 , 3 or 4 heteroaryl groups of 5-10 atoms independently selected from O, S or N heteroatoms), and, when R ^a and R ^b are connected to the same nitrogen atom, R ^a , R ^b , Together with the nitrogen atoms connected to them, a heterocyclic group with 3-8 atoms can also be optionally formed, wherein, the C _1-6 aliphatic, C _3-6 cycloalkyl, C _3-6 heterocyclic radical, C _6-10 aryl, heteroaryl of 5-10 atoms containing 1, 2, 3 or 4 heteroatoms independently selected from O, S or N, -(C _1-4 alkylene) -(C _3-6 cycloalkyl), -(C _1-4 alkylene)-(C _3-6 heterocyclyl), -(C _1-4 alkylene)-(C _6-10 aryl ), -(C _1-4 alkylene)-(heteroaryl group of 5-10 atoms containing 1, 2, 3 or 4 heteroatoms independently selected from O, S or N) and 3-8 Atoms of heterocyclyl are optionally substituted by 1, 2, 3 or 4 substituents independently selected from D, F, Cl, -CN, N ₃ , -OH, -NH ₂ , alkane Oxygen or alkylamino;

In special case, when Z is C _1-6 alkyl, -NR ^a R ^b cannot be -NH ₂ or -NMe ₂ .

In other embodiments, each of R ¹ , R ² , R ³ , R ⁴ , R ⁵ and R ⁶ is independently H or D.

In other embodiments, X is phenyl, and can be optionally represented by 1, 2, 3 or 4 independently selected from D, F, Cl, Br, C _1-3 alkyl or C _1-3 haloalkyl replaced by substituents.

In some other embodiments, Z is C _1-6 alkyl, C _3-6 cycloalkyl, C _3-6 heterocyclyl, C _5-12 fused bicyclyl, C _5-12 spiro bicyclyl, - (C _1-4 alkylene)-(C _3-6 heterocyclyl),-(C _1-4 alkylene)-(C _5-12 fused bicyclyl) or-(C _1-4 alkylene Base)-(C _5-12 spiro bicyclyl), wherein, the C _1-6 alkyl, C _3-6 cycloalkyl, C _3-6 heterocyclyl, C _5-12 fused bicyclyl, C _5-12 spirobicyclyl, -(C _1-4 alkylene) -(C _3-6 cycloalkyl), -(C _1-4 alkylene) -(C _3-6 heterocyclyl), - (C _1-4 alkylene)-(C _5-12 fused bicyclic group) or-(C _1-4 alkylene)-(C _5-12 spiro bicyclic group), optionally replaced by 1,2, Substituted by 3 or 4 substituents, the substituents are independently selected from D, F, Cl, Br, I, -CN, -NO ₂ , N ₃ , C _1-3 alkyl, -OR ^a , -NR ^a R ^b , and, C _5-12 fused bicyclyl, C _5-12 spiro bicyclyl, -(C _1-4 alkylene)-(C _5-12 fused bicyclyl) or -(C _1- Each ring in ₍₄ alkylene)-(C _5-12 spirobicyclyl) is independently carbocyclic or heterocyclic.

In other embodiments, each of R ^a and R ^b is independently H, C _1-3 alkyl, C _3-6 cycloalkyl or -(C _1-4 alkylene)-(C _3-6 cyclo Alkyl), and, when R ^a and R ^b are connected to the same nitrogen atom, R ^a , R ^b , together with the nitrogen atom connected to them, can also optionally form a heterocyclic group with 3-6 atoms , wherein, the C _1-3 alkyl group, C _3-6 cycloalkyl group, -(C _1-4 alkylene group)-(C _3-6 cycloalkyl group) and heterocyclic group with 3-6 atoms optionally substituted with 1, 2, 3 or 4 substituents independently selected from D, F, Cl, -OH, -NH ₂ , C _1-3 alkoxy or C _1-3 Alkylamino;

In special case, when Z is C _1-6 alkyl, -NR ^a R ^b cannot be -NH ₂ or -NMe ₂ .

In some other embodiments, Z is selected from any of the substructures shown in formulas (Z1) to (Z42):

or its stereoisomers, wherein:

n is 0, 1, 2 or 3;

each W and W' is independently -O- or -N(H)-;

The substructures represented by formulas (Z1) to (Z42) or their stereoisomers are optionally 1, 2, 3 or 4 independently selected from D, F, Cl, Br, I, -CN, -NO ₂ , N ₃ , C _1-3 alkyl, -OR ^a , -NR ^a R ^b substituents, wherein, when Z is the substructure shown in formula (Z25) ~ (Z32) or its stereoisomerism When it is a solid, -NR ^a R ^b cannot be -NH ₂ or -NMe ₂ .

In another aspect, the present invention relates to a pharmaceutical composition, which comprises the compound of the present invention, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, vehicle or a combination thereof.

In some embodiments, the pharmaceutical composition of the present invention further comprises an additional therapeutic agent selected from chemotherapy drugs, antiproliferative agents, drugs for treating atherosclerosis, and drugs for treating Medicines for pulmonary fibrosis or their combination.

In other embodiments, the pharmaceutical composition of the present invention, wherein the additional therapeutic agent involved is chlorambucil, melphalan, cyclophosphamide, ifosfamide (ifosfamide), busulfan, carmustine, lomustine, streptozocin, cisplatin, carboplatin, oxage Oxaliplatin, dacarbazine, temozolomide, procarbazine, methotrexate, fluorouracil, cytarabine, gemcitabine , mercaptopurine, fludarabine, vinblastine, vincristine, vinorelbine, paclitaxel, docetaxel, topotecan (topotecan), irinotecan, etoposide, trabectedin, dactinomycin, doxorubicin, epirubicin, Dow Daunorubicin, mitoxantrone, bleomycin, mitomycin, ixabepilone, tamoxifen, fluta Flutamide, gonadorelin analogues, megestrol, prednidone, dexamethasone, methylprednisolone, thalidomide ), interferon alfa, leucovorin, sirolimus, temsirolimus, everolimus, afatinib , alisertib, amuvatinib, apatinib, axitinib nib), bortezomib, bosutinib, brivanib, cabozantinib, cediranib, crenolanib, crizotinib, dabrafenib, dacomitinib, danusertib, dasatinib ( dasatinib), dovitinib, erlotinib, foretinib, ganetespib, gefitinib, ibrutinib, icotinib, imatinib, iniparib, lapatinib ( lapatinib), lenvatinib, linifanib, linsitinib, masitinib, momelotinib, motesanib, neratinib, nilotinib, niraparib, oprozomib, olaparib, prazole Pazopanib, pictilisib, ponatinib, quizartinib, regorafenib, rigosertib, rucaparib, ruxolitinib, saracatinib, saridegib, sorafenib, sunitinib, tasocitinib, telatinib, tivantinib, tivozanib, tofacitinib, trametinib, vandetanib, veliparib, vemurafenib, vismodegib, volasertib, alemtuzumab, bevacizumab, brentuximab vedotin, card Catumaxomab, cetuximab, denosumab, gemtuzumab, ipilimumab, nimotuzumab , ofatumumab, panitumumab, rituximab, tositumomab, trastuzumab, or combinations thereof.

On the other hand, the compounds or pharmaceutical compositions of the present invention can be used to prepare medicines for preventing, treating, treating or alleviating proliferative diseases in patients.

In some embodiments, the proliferative disease of the present invention is metastatic cancer, colon cancer, gastric adenocarcinoma, bladder cancer, breast cancer, kidney cancer, liver cancer, lung cancer, skin cancer, thyroid cancer, brain tumor, neck cancer, prostate cancer , pancreatic cancer, CNS (central nervous system) cancer, malignant glioma, myeloproliferative disease, atherosclerosis or pulmonary fibrosis.

In another aspect, the present invention relates to the use of the compound or pharmaceutical composition of the present invention to prepare a drug for inhibiting or regulating protein kinase activity in a biological sample, the use comprising contacting the biological sample with the compound of the present invention.

In some of these embodiments, the protein kinase described herein is a receptor tyrosine kinase.

In other embodiments, the receptor tyrosine kinase of the present invention is ALK, c-Met, or a combination thereof.

In another aspect, the present invention provides pharmaceutical compositions comprising compounds of the present invention, or stereoisomers, geometric isomers, tautomers, solvates, metabolites thereof, as inhibitors of tyrosine kinase receptors product, or a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable carrier, diluent, adjuvant, vehicle, or a combination thereof. In some embodiments, the pharmaceutical composition provided by the present invention comprises ALK receptor signaling response, c-Met receptor signaling response, or stereoisomers, geometric isomers, tautomers, solvates thereof , a metabolite, or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable carrier, diluent, adjuvant, vehicle, or a combination thereof. In other embodiments, the pharmaceutical compositions of the present invention further comprise additional therapeutic agents.

In another aspect, the invention relates to a method of inhibiting the activity of a protein tyrosine kinase comprising contacting a compound of the invention, or a pharmaceutical composition thereof, with said kinase. In some embodiments, the present invention relates to a method of inhibiting ALK receptor signaling, c-Met receptor signaling, the method comprising contacting a compound of the present invention, or a pharmaceutical composition thereof, with said receptor. In other embodiments, the inhibition of protein kinase receptor activity, particularly the activity of ALK or c-Met receptor signaling in a cell or multicellular organism, is inhibited. According to the method of the present invention, the method comprises administering to said multicellular organism a compound of the present invention or a pharmaceutical composition thereof. In some embodiments, the multicellular organism is a mammal. In other embodiments, the multicellular organism is a human. In some embodiments, the methods described herein further comprise contacting the kinase with an additional therapeutic agent.

In another aspect, the present invention relates to a method of inhibiting the proliferative activity of a cell, said method comprising contacting the cell with a therapeutically effective amount of a compound of the present invention or a pharmaceutical composition thereof capable of inhibiting proliferation. In some embodiments, the methods described herein further comprise contacting the cell with an additional therapeutic agent.

In another aspect, the present invention relates to a method of treating a cell proliferative disorder in a patient, said method comprising administering to the patient a therapeutically effective amount of a compound of the present invention or a pharmaceutical composition thereof. In some embodiments, the methods described herein further comprise the administration of an additional therapeutic agent.

In another aspect, the present invention relates to a method of inhibiting tumor growth in a patient, said method comprising administering to the patient a therapeutically effective amount of a compound of the present invention or a pharmaceutical composition thereof. In some embodiments, the methods described herein further comprise the administration of an additional therapeutic agent.

In another aspect, the present invention relates to processes for the preparation, isolation and purification of compounds encompassed by formula (I).

Experimental results show that the compound provided by the invention exhibits good half-life and good pharmacokinetic properties, has good inhibitory effect on ALK and c-Met, and also has good inhibitory effect on tumor growth.

The preceding description merely outlines certain aspects of the invention, but is not intended to be limiting. These and other aspects will be described in more detail and more fully below.

Detailed ways

Definitions and General Terms

The present invention will list in detail the literature corresponding to the determined and embodied content, and the examples are all accompanied by illustrations of structural formulas and chemical formulas. The present invention is intended to cover all alternatives, modifications and equivalents which may be included within the scope of the present invention as defined by the claims. One skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in the practice of the present invention. The invention is in no way limited to the description of the methods and materials. There are many documents and similar materials that differ from or contradict the present application, including but in no way limited to the definitions of terms, term usage, described techniques, or the scope of control like the present application.

Unless otherwise indicated, the following definitions apply to this invention:

For purposes of the present invention, chemical elements are defined according to the Periodic Table of the Elements, CAS Edition and Handbook of Chemicals, 75, ^th Ed, 1994. In addition, general principles of organic chemistry can be found in "Organic Chemistry," Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry," by Michael B. Smith and Jerry March, John Wiley & Sons, New York: 2007, so all The contents are incorporated with references.

As described in the present invention, the compounds of the present invention can be optionally substituted by one or more substituents, such as the above general formula compounds, or as specific examples in the examples, subclasses, and included in the present invention A class of compounds. It should be understood that the term "optionally substituted" and the term "substituted or unsubstituted" are used interchangeably. "Optionally" or "optionally" means that the subsequently described event or circumstance can but need not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example: "Heterocyclyl is optionally substituted by 1 or 2 groups independently selected from D, F" means that D, F may but not necessarily exist, and the description includes wherein the heterocyclyl is substituted by 1 or 2 The case where two groups independently selected from D and F are substituted, and the case where the heterocyclic group is unsubstituted.

In general, the term "substituted" or "substituted" means that one or more hydrogen atoms in a given structure are replaced by a particular substituent. Unless otherwise indicated, an optionally substituted group may have a substituent at each substitutable position of the group. When more than one position in a given formula can be substituted by one or more substituents selected from a particular group, then the substituents can be substituted at each position the same or differently. The substituents mentioned therein can be, but are not limited to, deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, azido, alkyl, haloalkyl, alkenyl, alkynyl, hydroxyalkyl , alkoxyalkyl, aminoalkyl, alkylaminoalkyl, cyanoalkyl, hydroxyl, alkoxy, mercapto, alkylthio, amino, alkylamino, aminoacyl, alkylaminoacyl, aryl, heteroaryl base and so on.

The term "aliphatic" or "aliphatic group", as used herein, means a straight (unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is fully saturated or contains one or more degrees of unsaturation. Unless otherwise specified, aliphatic groups contain 1-20 carbon atoms. In some embodiments, the aliphatic group contains 1-10 carbon atoms, and in other embodiments, the aliphatic group contains 1-8 carbon atoms. In some other embodiments, the aliphatic group contains 1-6 carbon atoms, in some other embodiments, the aliphatic group contains 1-4 carbon atoms, in some other embodiments, the aliphatic group contains 1-3 carbon atoms, and in some other embodiments, the aliphatic group contains 1-2 carbon atoms. Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, or alkynyl groups. For example, -(C ₁ -C ₆ )aliphatic, including unbranched or branched, unsubstituted or suitably substituted -(C ₁ -C ₆ )alkyl, -(C ₂ -C ₆ )alkenyl , or -(C ₂ -C ₆ )alkynyl. Examples of such include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, ethylene, propylene, butene, 2-butene, acetylene, propyne , butyne, 2-butyne, etc., and the aliphatic can be substituted or unsubstituted, where the substituents can be, but not limited to, deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, azido, alkyl, haloalkyl, alkenyl, alkynyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, alkylaminoalkyl, cyanoalkyl, hydroxy, alkoxy, mercapto, alkane Thio, amino, alkylamino, aminoacyl, alkylaminoacyl, aryl, heteroaryl, etc.

The term "alkyl" or "alkyl group" used in the present invention means a saturated linear or branched monovalent hydrocarbon atomic group containing 1-20 carbon atoms. Wherein said alkyl groups may be independently and optionally substituted by one or more substituents. Unless otherwise specified, the alkyl group contains 1-20 carbon atoms, some embodiments are, the alkyl group contains 1-10 carbon atoms, and in other embodiments, the alkyl group contains 1-8 carbon atom, some other embodiments are that the alkyl group contains 1-6 carbon atoms, some other embodiments are that the alkyl group contains 1-4 carbon atoms, some other embodiments are that the alkyl group contains 1-3 carbon atoms, and in other embodiments, the alkyl group contains 1-2 carbon atoms.

Examples of alkyl groups include, but are not limited to, methyl (Me, -CH ₃ ), ethyl (Et, -CH ₂ CH ₃ ), n-propyl (n-Pr, -CH ₂ CH ₂ CH ₃ ), isopropyl (i-Pr, i-propyl, -CH(CH ₃ ) ₂ ), n-butyl (n-Bu, n-butyl, -CH ₂ CH ₂ CH ₂ CH ₃ ), isobutyl ( i-Bu, i-butyl, -CH ₂ CH(CH ₃ ) ₂ ), sec-butyl (s-Bu, s-butyl, -CH(CH ₃ )CH ₂ CH ₃ ), tert-butyl (t-Bu , t-butyl, -C(CH ₃ ) ₃ ), n-pentyl (n-pentyl, -CH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ), 2-pentyl (-CH(CH ₃ )CH ₂ CH ₂ CH ₃ ), 3-pentyl (-CH(CH ₂ CH ₃ ) ₂ ), 2-methyl-2-butyl (-C(CH ₃ ) ₂ CH ₂ CH ₃ ), 3-methyl-2- Butyl (-CH(CH ₃ ) CH(CH ₃ ) ₂ ), 3-methyl-1-butyl (-CH ₂ CH ₂ CH(CH ₃ ) ₂ ), 2-methyl-1-butyl ( -CH ₂ CH(CH ₃ )CH ₂ CH ₃ ), n-hexyl (-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ), 2-hexyl (-CH(CH ₃ )CH ₂ CH ₂ CH ₂ CH ₃ ), 3-hexyl (-CH(CH ₂ CH ₃ )(CH ₂ CH ₂ CH ₃ )), 2-methyl-2-pentyl (-C(CH ₃ ) ₂ CH ₂ CH ₂ CH ₃ ), 3 -Methyl-2-pentyl (-CH(CH ₃ )CH(CH ₃ )CH ₂ CH ₃ ), 4-methyl-2-pentyl (-CH(CH ₃ )CH ₂ CH(CH ₃ ) ₂ ), 3-methyl-3-pentyl (-C(CH ₃ )(CH ₂ CH ₃ ) ₂ ), 2-methyl-3-pentyl (-CH(CH ₂ CH ₃ )CH(CH ₃ ) ₂ ), 2,3-dimethyl-2-butyl (-C(CH ₃ ) ₂ CH(CH ₃ ) ₂ ), 3,3-dimethyl-2-butyl (-CH(CH ₃ ) C(CH ₃ ) ₃ ), n-heptyl, n-octyl, etc. And the alkyl group can be substituted or unsubstituted, where the substituents can be, but not limited to, deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, azido, alkyl, haloalkyl, alkenyl radical, alkynyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, alkylaminoalkyl, cyanoalkyl, hydroxy, alkoxy, mercapto, alkylthio, amino, alkylamino, aminoacyl, alkyl Aminoacyl, aryl, heteroaryl, etc.

As used herein, the term "alkyl" and its prefix "alk" include both straight and branched saturated carbon chains.

The term "alkylene" used in the present invention means a saturated divalent hydrocarbon group obtained by eliminating two hydrogen atoms from a straight-chain or branched-chain saturated hydrocarbon. Unless otherwise specified, the alkylene group contains 1-10 carbon atoms, in some embodiments, the alkylene group contains 1-6 carbon atoms, and in other embodiments, the alkylene group contains 1 - 4 carbon atoms, in other embodiments the alkylene group contains 1-3 carbon atoms, in other embodiments the alkylene group contains 1-2 carbon atoms. Examples of alkylene groups include, but are _not limited _to , methylene ( _-CH2- ), ethylene _{( -CH2CH2-} ), propylene ( _-CH2CH2CH2- ), Ethyl (-CH(CH ₃ )-), Isopropyl (-CH(CH ₃ )CH ₂ -) and the like. The alkylene group may be further substituted selected from, but not limited to, deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, azido, alkyl, haloalkyl, alkenyl, alkyne hydroxyalkyl, alkoxyalkyl, aminoalkyl, alkylaminoalkyl, cyanoalkyl, hydroxy, alkoxy, mercapto, alkylthio, amino, alkylamino, aminoacyl, alkylaminoacyl, Aryl, heteroaryl, etc.

The term "alkenyl" means 2-12 carbon atoms, or 2-8 carbon atoms, or 2-6 carbon atoms, or 2-4 carbon atoms straight-chain or branched monovalent hydrocarbon groups, wherein at least one The position is in an unsaturated state, that is, one CC is a sp ² double bond, and the alkenyl group can be independently and optionally replaced by one or more substituents described in the present invention, including groups with "anti"" Orientation of "normal" or "E" and "Z", where specific examples include, but are not limited to, vinyl (-CH=CH ₂ ), allyl (-CH ₂ CH=CH ₂ ) and the like.

The term "alkynyl" means 2-12 carbon atoms, or 2-8 carbon atoms, or 2-6 carbon atoms, or 2-4 carbon atoms linear or branched monovalent hydrocarbon groups, wherein at least one position It is an unsaturated state, that is, one CC is a sp triple bond, wherein the hydrocarbyl group can be independently and optionally replaced by one or more substituents described in the present invention, specific examples include, but are not limited to, ethynyl ( -C≡CH), propargyl (-CH ₂ C≡CH) and the like.

The term "haloalkyl", "haloalkenyl" or "haloalkoxy" means an alkyl, alkenyl or alkoxy group substituted with one or more halogen atoms, wherein alkenyl and alkoxy Such examples include, but are not limited to, trifluoromethyl, trifluoromethoxy, etc. within the meanings described herein.

The term "carbocycle", "carbocyclyl" or "carbocyclic" means a monovalent or polyvalent, non-aromatic, saturated or partially unsaturated, monocyclic, bicyclic or tricyclic ring containing 3 to 12 carbon atoms . Suitable cycloaliphatic groups include, but are not limited to, cycloalkyl, cycloalkenyl and cycloalkynyl. Examples of cycloaliphatic groups further include, but are by no means limited to, cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopentyl-1-enyl, 1-cyclopentyl-2-enyl, 1-cyclopentyl-3-enyl, cyclohexyl, 1-cyclohexyl-1-enyl, 1-cyclohexyl-2-enyl, 1-cyclohexyl-3-enyl, cyclohexadienyl, etc. And the "cycloalkyl" refers to a monovalent or polyvalent, saturated, monocyclic ring containing 3-12 carbon atoms. And the cycloalkyl group can be further substituted, the substituent is selected from, but not limited to, deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, azido, alkyl, haloalkyl, alkenyl, Alkynyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, alkylaminoalkyl, cyanoalkyl, hydroxy, alkoxy, mercapto, alkylthio, amino, alkylamino, aminoacyl, alkylaminoacyl , aryl, heteroaryl, etc.

The term "cycloalkylalkylene" means that an alkyl group may be substituted by one or more cycloalkyl groups, wherein alkyl and cycloalkyl groups have the meanings described herein. In some of these embodiments, a cycloalkylalkylene group refers to a "lower cycloalkylalkylene" group, ie a cycloalkyl group attached to a C _1-6 alkyl group. In some other embodiments, the cycloalkyl group is attached to a C _1-4 alkyl group. In some other embodiments, the cycloalkyl group is attached to a C _1-3 alkyl group. Such examples include, but are not limited to, cyclopropylethyl, cyclopentylmethyl, cyclohexylmethyl, and the like. Cycloalkylalkylene can be further substituted, the substituents can be, but are not limited to, deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, azido, alkyl, haloalkyl, alkenyl radical, alkynyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, alkylaminoalkyl, cyanoalkyl, hydroxy, alkoxy, mercapto, alkylthio, amino, alkylamino, aminoacyl, alkyl Aminoacyl, aryl, heteroaryl, etc.

The terms "heterocycle", "heterocyclyl" or "heterocyclic" are used interchangeably herein to refer to monocyclic or bicyclic ring systems in which one or more atoms of the ring are independently optionally replaced by heteroatoms , the ring can be fully saturated or contain one or more degrees of unsaturation, but is never aromatic, with only one point of attachment to the rest of the molecule. One or more ring hydrogen atoms are independently optionally substituted with one or more substituents described herein. In some embodiments, a "heterocycle", "heterocyclyl" or "heterocyclic" group is a 3-6 membered monocyclic ring (2-5 carbon atoms and selected from N, O, P, S 1-3 heteroatoms, where S or P is optionally substituted by one or more oxygen atoms to obtain groups like SO, SO ₂ , PO, PO ₂ , when the ring is a three-membered ring , where there is only one heteroatom), or a 7-10 membered bicyclic ring (4-9 carbon atoms and 1-3 heteroatoms selected from N, O, P, S, where S or P is optionally replaced by a Or more oxygen atoms are substituted to get groups like SO, SO ₂ , PO, PO ₂ ).

A heterocyclic group may be a carbon group or a heteroatom group. Examples of heterocyclic rings include, but are not limited to, pyrrolidinyl, tetrahydrofuryl, dihydrofuryl, tetrahydrothiophenyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidinyl, Morpholinyl, Thiomorpholinyl, Thioxanyl, Piperazinyl, Homopiperazinyl, Azetidinyl, Oxetanyl, Thietanyl, Homopiperidinyl, Glycidyl Base, azepanyl, oxepinyl, thiepanyl, oxazepinyl, diazepinyl, thiazepinyl, 2-pyrrolinyl, 3-pyrrolinyl, Indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolyl, pyrazolinyl, dithianyl, dithianyl, di Hydrogen thienyl, pyrazolidinyl imidazolinyl, imidazolidinyl, 1,2,3,4-tetrahydroisoquinolinyl. Examples of the heterocyclic group also include a pyrimidinedione group and a 1,1-dioxothiomorpholinyl group in which two ring carbon atoms are substituted with oxygen (=O). And the heterocyclic group is independently and optionally substituted by one or more substituents described in the present invention, wherein the substituents can be, but not limited to, deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro , azido, alkyl, haloalkyl, alkenyl, alkynyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, alkylaminoalkyl, cyanoalkyl, hydroxy, alkoxy, mercapto, Alkylthio, amino, alkylamino, aminoacyl, alkylaminoacyl, aryl, heteroaryl, and the like.

The term "heterocyclylalkylene" means that an alkyl group may be substituted by one or more heterocyclyl groups, wherein the alkyl and heterocyclyl groups have the meanings described herein. In some of these embodiments, a heterocyclylalkylene group refers to a "lower heterocyclylalkylene" group, ie a heterocyclyl group attached to a C _1-6 alkyl group. In some other embodiments, the heterocyclyl group is attached to a C _1-4 alkyl group. Such examples include, but are not limited to, 2-pyrrolidinylethyl and the like. Heterocyclylalkylene can be further substituted, the substituents can be, but not limited to, deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, azido, alkyl, haloalkyl, alkenyl , alkynyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, alkylaminoalkyl, cyanoalkyl, hydroxy, alkoxy, mercapto, alkylthio, amino, alkylamino, aminoacyl, alkylamino Acyl, aryl, heteroaryl, etc.

The term "heteroatom" means one or more of O, S, N, P, and Si, including forms in any oxidation state of N, S, and P; forms of primary, secondary, and tertiary amines, and quaternary ammonium salts; or nitrogen in heterocycles The form in which the hydrogen on the atom is substituted, for example, N (like N in 3,4-dihydro-2H-pyrrolyl), NH (like NH in pyrrolidinyl) or NR (like N-substituted pyrrolidine NR in the base).

The term "halogen" refers to F, Cl, Br or I.

The term "H" denotes a single hydrogen atom. Such atomic groups can be linked to other groups, such as oxygen atoms, to form hydroxyl groups.

The term "D" or " ^2H " denotes a single deuterium atom. One such group is attached to a methyl group to form a mono-deuteromethyl group ( _-CDH2 ), two deuterium atoms are attached to a methyl group to form a bis-deuteromethyl group ( _-CD2H ), and three The deuterium atom is attached to a methyl group to form tri-deuteromethyl (-CD ₃ ).

The term " _N3 " denotes an azide structure. This group can be linked with other groups, for example, with a methyl group, it can form azidomethane (methyl azide, MeN ₃ ); and with a phenyl group, it can form a phenyl Azide (PhN ₃ ).

The term "aryl" may be used alone or as part of "aralkyl", "aralkoxy" or "aryloxyalkyl" and denotes monocyclic, bicyclic, and A tricyclic carbocyclic ring system wherein at least one ring system is aromatic, wherein each ring system contains 3-7 membered rings, and has only one point of attachment to the rest of the molecule. The term "aryl" is used interchangeably with the term "aromatic ring", which may include phenyl, naphthyl and anthracene. And the aryl group can be substituted or unsubstituted, wherein the substituents can be, but not limited to, deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, azido, alkyl, haloalkyl, Alkenyl, Alkynyl, Hydroxyalkyl, Alkoxyalkyl, Aminoalkyl, Alkaminoalkyl, Cyanoalkyl, Hydroxy, Alkoxy, Mercapto, Alkylthio, Amino, Alkamino, Aminoacyl , alkylaminoacyl, aryl, heteroaryl, etc.

The term "arylalkylene" means that an alkyl group may be substituted by one or more aryl groups, wherein the alkyl and aryl groups have the meanings described herein, some of which are, aryl An alkylene group refers to a "lower arylalkylene" group, ie, an aryl group attached to a C _1-6 alkyl group. In some other embodiments, an arylalkylene group refers to a "phenylene alkylene" containing a C _1-4 alkyl group. Specific examples thereof include benzyl, diphenylmethyl, phenethyl and the like. Arylalkylene may be further substituted, the substituents may be, but are not limited to, deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, azido, alkyl, haloalkyl, alkenyl , alkynyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, alkylaminoalkyl, cyanoalkyl, hydroxy, alkoxy, mercapto, alkylthio, amino, alkylamino, aminoacyl, alkylamino Acyl, aryl, heteroaryl, etc.

The term "heteroaryl" which may be used alone or as part of "heteroaralkyl" denotes monocyclic, bicyclic, and tricyclic ring systems containing 5-14 ring atoms, wherein at least one ring system is aromatic , and at least one ring system comprises one or more heteroatoms, wherein each ring system comprises a 5-7 membered ring and only one point of attachment is to the rest of the molecule. The term "heteroaryl" may be used interchangeably with the terms "heteroaromatic ring" or "heteroaromatic". And the heteroaryl group can be substituted or unsubstituted, wherein the substituents can be, but not limited to, deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, azido, alkyl, haloalkyl , alkenyl, alkynyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, alkylaminoalkyl, cyanoalkyl, hydroxy, alkoxy, mercapto, alkylthio, amino, alkylamino, amino Acyl, alkylaminoacyl, aryl, heteroaryl, etc.

In some other embodiments, aromatic heterocycles include, but are not limited to, the following monocycles: 2-furyl, 3-furyl, N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5- Imidazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, N-pyrrolyl, 2-pyrrole Base, 3-pyrrolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, pyridazinyl (such as 3-pyridazinyl), 2 -thiazolyl, 4-thiazolyl, 5-thiazolyl, tetrazolyl (such as 5-tetrazolyl), triazolyl (such as 2-triazolyl and 5-triazolyl), 2-thienyl, 3 - Thienyl, pyrazolyl (such as 2-pyrazolyl), isothiazolyl, 1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl Azolyl, 1,2,3-triazolyl, 1,2,3-thiodiazolyl, 1,3,4-thiodiazolyl, 1,2,5-thiodiazolyl, pyr Azinyl, 1,3,5-triazinyl; also includes, but is by no means limited to, the following bicyclic rings: benzimidazolyl, benzofuryl, benzothienyl, indolyl (such as 2-indole base), purinyl, quinolinyl (such as 2-quinolyl, 3-quinolyl, 4-quinolyl), and isoquinolyl (such as 1-isoquinolyl, 3-isoquinolyl or 4-isoquinolinyl).

The term "heteroarylalkylene" means that an alkyl group may be substituted by one or more heteroaryl groups, wherein the alkyl and heteroaryl groups have the meanings described herein, some of which Yes, a heteroarylalkylene group refers to a "lower heteroarylalkylene" group, ie a heteroaryl group attached to a C _1-6 alkyl group. In some other embodiments, the heteroaryl group is attached to a C _1-4 alkyl group. Specific examples thereof include 2-pyridylmethyl, 3-furylethyl and the like. The heteroarylalkylene can be further substituted, the substituents can be, but not limited to, deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, azido, alkyl, haloalkyl, chain alkenyl, alkynyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, alkylaminoalkyl, cyanoalkyl, hydroxy, alkoxy, mercapto, alkylthio, amino, alkylamino, aminoacyl, Alkaminoacyl, aryl, heteroaryl, etc.

The term "carboxy", whether used alone or in combination with other terms, such as "carboxyalkyl", means -CO ₂ H; the term "carbonyl", whether used alone or in combination with other terms, such as "aminocarbonyl" or ""Acyloxy" means -(C=O)-.

The term "alkylthio" includes a C _1-10 linear or branched alkyl group connected to a divalent sulfur atom, and some of the embodiments are that the alkylthio group is a lower C _1-3 alkylthio group, Such examples include, but are not limited to, methylthio ( _CH3S- ).

The term "alkoxy" as used herein refers to an alkyl group, as defined herein, attached to the main carbon chain through an oxygen atom ("alkoxy"), examples of which include, but do not Not limited to methoxy, ethoxy, propoxy, butoxy and the like.

The term "alkoxyalkyl" means that an alkyl group is substituted by one or more alkoxy groups, wherein the alkyl group and the alkoxy group have the meanings described herein, examples of which include , but not limited to methoxymethyl, ethoxymethyl, ethoxyethyl, etc.

The term "alkylamino" or "alkylamino" includes "N-alkylamino" and "N,N-dialkylamino", wherein the amino groups are each independently substituted with one or two alkyl groups. In some embodiments, alkylamino is a lower alkylamino group with one or two C _1-6 alkyl groups attached to a nitrogen atom. In some other embodiments, the alkylamino is a C _1-3 lower alkylamino group. Suitable alkylamino groups may be mono- or di-alkylamino, examples of which include, but are not limited to, N-methylamino, N-ethylamino, N,N-dimethylamino, N,N- -Diethylamino and the like.

The term "arylamino" means that an amino group is substituted by one or two aryl groups, examples of which include, but are not limited to, N-phenylamino. In some embodiments, the aromatic ring on the arylamino group can be further substituted.

The term "hydroxyalkyl" includes C _1-10 straight or branched chain alkyl groups substituted with one or more hydroxy groups. In some embodiments, hydroxyalkyl is C _1-6 "lower hydroxyalkyl" substituted with one or more hydroxy groups, examples of which include, but are not limited to, hydroxymethyl, hydroxy Ethyl, hydroxypropyl, hydroxybutyl and hydroxyhexyl.

The term "aminoalkyl" includes C _1-10 straight or branched chain alkyl groups substituted with one or more amino groups. In some embodiments, aminoalkyl is C _1-6 "lower aminoalkyl" substituted with one or more amino groups, examples of which include, but are not limited to, aminomethyl, amino Ethyl, aminopropyl, aminobutyl and aminohexyl.

The term "cyanoalkyl" includes C _1-10 straight or branched chain alkyl groups substituted with one or more cyano groups. In some embodiments, cyanoalkyl is C _1-6 "lower cyanoalkyl" substituted with one or more cyano groups, examples of which include, but are not limited to, cyano Methyl, cyanoethyl, cyanopropyl, cyanobutyl and cyanohexyl.

The term "alkaminoalkyl" includes alkyl groups substituted with alkylamino groups. In some embodiments, the alkylaminoalkyl is a C _1-6 lower alkylaminoalkyl. In some other embodiments, the alkylaminoalkyl is C _1-3 lower alkylaminoalkyl. Suitable alkylaminoalkyl groups may be mono- or di-alkyl substituted, examples of which include, but are not limited to, N-methylaminomethyl, N,N-dimethylaminoethyl, N,N-diethylaminomethyl and so on.

As used herein, the term "unsaturated" means that a moiety contains one or more degrees of unsaturation.

The term "comprising" is an open expression, that is, it includes the content specified in the present invention, but does not exclude other content.

The term "fused bicyclic", "fused ring", "fused bicyclic group" or "fused cyclic group", as shown in Figures a-c, means between two five-membered rings (formula a), two six-membered Between rings (formula b), and between a five-membered ring and a six-membered ring (formula c) share a bridged ring system with a C-C bond. The system may contain isolated or conjugated unsaturated bonds, but the core ring structure does not include aromatic or heteroaromatic rings (substituents may be aromatic). Each ring in a fused bicyclic ring can be carbocyclic or heterocyclic.

Examples of fused bicyclic rings include, but are not limited to, hexahydrofuro[2,3-b]furan-3-yl, 3a-fluorohexahydrofuro[2,3-b]furan-3-yl, (3aS ,6aR)-3a,6a-dideuterium hexahydrofuro[2,3-b]furan-3-yl, 5,5-difluorohexahydrofuro[2,3-b]furan-3-yl, 4,4-Difluorohexahydrofuro[2,3-b]furan-3-yl, (3S)-6-hydroxyhexahydrofuro[3,2-b]furan-3-yl, (6R) -6-fluorohexahydrofuro[3,2-b]furan-3-yl, 3-deuterium hexahydrofuro[3,2-b]furan-3-yl, 5,5-dideuterium hexahydrofuran And[3,2-b]furan-3-yl, 6,6-difluorohexahydrofuro[3,2-b]furan-3-yl, (3aR,6aR)-3a,6a-dideuterium hexa Hydrofuro[3,2-b]furan-3-yl, octahydrocyclopenta[c]pyrrol-5-yl, (3aR,6aS)-3a,6a-dideuteroctahydrocyclopenta[ c]pyrrol-5-yl, 3a-fluorodideuteroctahydrocyclopenta[c]pyrrol-5-yl, 5-deuterium octahydrocyclopenta[c]pyrrol-5-yl, 1,1- Dideuterium octahydrocyclopenta[c]pyrrol-5-yl, 5-aminooctahydropentalen-2-yl, 5-deuterium-5-aminooctahydropentalen-2-yl ,etc. Fused bicyclic groups can be substituted or unsubstituted, where substituents can be, but are not limited to, deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, azido, alkyl, haloalkyl, chain alkenyl, alkynyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, alkylaminoalkyl, cyanoalkyl, hydroxy, alkoxy, mercapto, alkylthio, amino, alkylamino, aminoacyl, Alkaminoacyl, aryl, heteroaryl, etc.

The term "fused bicyclylalkylene" means that an alkyl group is substituted by one or more fused bicyclyl groups, wherein the alkyl group and the fused bicyclyl group have the meanings described herein, such that Examples include, but are not limited to, hexahydrofuro[2,3-b]furan-3-ylmethyl, hexahydrofuro[2,3-b]furan-3-yldideuteromethyl, (3aR, 6aS)-3a,6a-dideuterohexahydrofuro[2,3-b]furan-3-ylmethyl and the like. Fused bicycloalkylene can be substituted or unsubstituted, where substituents can be, but are not limited to, deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, azido, alkyl, haloalkane radical, alkenyl, alkynyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, alkylaminoalkyl, cyanoalkyl, hydroxy, alkoxy, mercapto, alkylthio, amino, alkylamino, Aminoacyl, alkylaminoacyl, aryl, heteroaryl, etc.

The term "spirocyclyl", "spirocycle", "spirobicyclyl" or "spirobicyclyl" means that one ring is derived from a specific cyclic carbon on another ring, e.g., as described by formula d below, a saturated The bridged ring system (rings B and B') is called a "fused bicyclic ring", whereas ring A and ring B share a carbon atom in two saturated ring systems, which are called "spiro rings" or "spiro rings". Double Ring". Each ring in a spirocycle is independently carbocyclic or heterocyclic. Such examples include, but are not limited to, 5-azaspiro[2.4]heptan-5-yl, (R)-4-azaspiro[2.4]heptan-6-yl, and the like. Spirobicyclyl can be substituted or unsubstituted, where substituents can be, but are not limited to, deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, azido, alkyl, haloalkyl, alkenyl radical, alkynyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, alkylaminoalkyl, cyanoalkyl, hydroxy, alkoxy, mercapto, alkylthio, amino, alkylamino, aminoacyl, alkyl Aminoacyl, aryl, heteroaryl, etc.

The term "spirobicyclylalkylene" means that an alkyl group is substituted by one or more spirobicyclyl groups, wherein the alkyl group and the spirobicyclyl group have the meanings described herein. Spirobicyclylalkylene can be substituted or unsubstituted, where substituents can be, but are not limited to, deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, azido, alkyl, haloalkyl , alkenyl, alkynyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, alkylaminoalkyl, cyanoalkyl, hydroxy, alkoxy, mercapto, alkylthio, amino, alkylamino, amino Acyl, alkaminoacyl, aryl, heteroaryl, etc.

As described in the present invention, the substituents draw a bond to the central ring to form a ring system (as shown in the figure below) which means that any substitutable position of the substituent on the ring can be substituted. For example, formula e represents any position that may be substituted on ring B, as shown in formula f.

Unless otherwise indicated, the structural formulas described in the present invention include all isomeric forms (such as enantiomers, diastereomers, and geometric isomerism (or conformational isomerism)): for example, those containing asymmetric centers R, S configuration, double bond (Z), (E) isomers, and (Z), (E) conformational isomers. Accordingly, individual stereochemical isomers of the compounds of the present invention or mixtures thereof as enantiomers, diastereomers, or geometric isomers (or conformational isomers) are within the scope of the present invention.

The term "tautomer" or "tautomeric form" as used in the present invention means that structural isomers with different energies can cross a low energy barrier and thus interconvert. For example, proton tautomerization (ie, prototropism) includes interconversions via migration of a proton, such as keto-enol tautomerization and imine-enamine isomerization. Valence tautomers include interconversion by recombination of some of the bonding electrons.

Unless otherwise indicated, all tautomeric forms of the compounds of the invention are included within the scope of the invention. In addition, unless otherwise indicated, the structural formulas of the compounds described herein include enriched isotopes of one or more different atoms.

The term "prodrug" used in the present invention means that a compound is transformed into a compound represented by formula (I) in vivo. Such conversion is effected by prodrug hydrolysis in blood or enzymatic conversion in blood or tissue to the parent structure. The prodrug compound of the present invention can be an ester. In the existing invention, the ester can be used as a prodrug including phenyl esters, aliphatic (C _1-24 ) esters, acyloxymethyl esters, carbonates , carbamates and amino acid esters. For example, a compound of the present invention contains an OH group, which can be acylated to give the compound in prodrug form. Other prodrug forms include phosphate esters, eg, phosphorylated parent hydroxyl groups. A complete discussion of prodrugs can be found in the following literature: T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, Vol. 14 of the A. CSS Symposium Series, Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, J. Rautio et al, Prodrugs: Design and Clinical Applications, Nature Review Drug Discovery, 2008, 7, 255-270, and SJHecker et al, Prodrugs of Phosphates and Phosphonates, Journal of Medicinal Chemistry 1, 2008, 2328-2345, each of which is hereby incorporated by reference.

"Metabolite" refers to a product obtained through metabolism of a specific compound or its salt in vivo. Metabolites of a compound can be identified by techniques known in the art, and their activity can be characterized using assays as described herein. Such products can be obtained by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, degreasing, enzymatic cleavage and the like of the administered compound. Accordingly, the invention includes metabolites of the compounds, including metabolites produced by contacting a compound of the invention with a mammal for a substantial period of time.

Definitions of stereochemistry in the present invention and use of conventions are generally referred to in the following documents: S.P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S. ., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994. The compounds of the present invention may contain asymmetric centers or chiral centers and thus exist as different stereoisomers. All stereoisomeric forms of the compounds of the present invention, including but not limited to, diastereomers, enantiomers, atropisomers, and mixtures thereof, such as racemic mixtures, constitute the present invention part. Many organic compounds exist in optically active forms, that is, they have the ability to rotate the plane of plane-polarized light. When describing optically active compounds, the prefixes D, L or R, S are used to indicate the absolute configuration of the molecular chiral center. The prefixes d, l or (+), (-) are used to name the symbol of the compound's plane polarized light rotation, (-) or l means that the compound is left-handed, and the prefix (+) or d means that the compound is right-handed. The chemical structures of these stereoisomers are the same, but their three-dimensional structures are different. A particular stereoisomer may be an enantiomer, and a mixture of isomers is often referred to as an enantiomeric mixture. A 50:50 mixture of enantiomers is known as a racemic mixture or racemate, which can result in no stereoselectivity or stereospecificity during a chemical reaction. The terms "racemic mixture" and "racemate" refer to an equimolar mixture of two enantiomers, devoid of optical activity.

The "pharmaceutically acceptable salt" used in the present invention refers to organic and inorganic salts of the compounds of the present invention. Pharmaceutically acceptable salts are well known in the art, as described in SM Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66:1-19, 1977. Pharmaceutically acceptable non-toxic acid salts include, but are not limited to, inorganic acid salts formed by reaction with amino groups such as hydrochloride, hydrobromide, phosphate, sulfate, perchlorate, and organic acid salts such as acetate, oxalate, maleate, tartrate, citrate, succinate, malonate, or other methods such as ion exchange methods recorded in books and literature these salts. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, besylate, benzoate, bisulfate, borate, butyrate, camphorate Salt, camphorsulfonate, cyclopentylpropionate, digluconate, lauryl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate Salt, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, Malate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, palmitate, pamoate, pectate, persulfate, 3 - Phenylpropionate, picrate, pivalate, propionate, stearate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc. Salts obtained with appropriate bases include alkali metal, alkaline earth metal, ammonium and N ⁺ (C _1-4 alkyl) ₄ salts. The present invention also contemplates the quaternary ammonium salts of any compound containing an N group. Water-soluble or oil-soluble or dispersed products can be obtained by quaternization. Alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Pharmaceutically acceptable salts further include suitable, non-toxic ammonium, quaternary ammonium salts and amine cations formed as counterions, such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, C _{1 -8} sulfonates and aromatic sulfonates.

A "solvate" of the present invention refers to an association of one or more solvent molecules with a compound of the present invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethylsulfoxide, ethyl acetate, acetic acid, aminoethanol. The term "hydrate" refers to an association of solvent molecules with water.

The term "protecting group" or "PG" refers to a substituent that reacts with another functional group, usually to block or protect specific functionality. For example, "amino-protecting group" refers to a substituent attached to the amino group to block or protect the functionality of the amino group in the compound. Suitable amino-protecting groups include acetyl, trifluoroacetyl, tert-butoxycarbonyl (BOC), benzyloxycarbonyl (CBZ) and 9-fluorenylmethyleneoxycarbonyl (Fmoc). Similarly, a "hydroxyl protecting group" refers to a substituent of a hydroxy group used to block or protect the functionality of the hydroxy group, suitable protecting groups include acetyl and silyl groups. "Carboxyl protecting group" refers to the substituent of carboxyl to block or protect the functionality of carboxyl. General carboxyl protecting groups include -CH ₂ CH ₂ SO ₂ Ph, cyanoethyl, 2-(trimethylsilane base) ethyl, 2-(trimethylsilyl)ethoxymethyl, 2-(p-toluenesulfonyl)ethyl, 2-(p-nitrobenzenesulfonyl)ethyl, 2-(diphenyl phosphino)ethyl, nitroethyl, etc. For a general description of protecting groups, reference can be made to: T W. Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, New York, 1991; and PJ Kocienski, Protecting Groups, Thieme, Stuttgart, 2005.

compound

The heterocyclic compound involved in the present invention, its pharmaceutically acceptable salt, and its pharmaceutical preparation have potential application in the treatment of diseases or diseases regulated by tyrosine kinase receptors, especially ALK and c-Met receptors. In particular, the present invention relates to a compound represented by formula (I):

Or its stereoisomer, geometric isomer, tautomer, nitrogen oxide, solvate, metabolite, pharmaceutically acceptable salt or its prodrug.

Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , X and Z have the following meanings:

Each of R ¹ , R ² , R ³ , R ⁴ , R ⁵ and R ⁶ is independently H, D or F, and in some embodiments, may be independently H or D.

X is a C _6-10 aryl group or a heteroaryl group of 5-10 atoms containing 1, 2, 3 or 4 heteroatoms independently selected from O, S or N, wherein the C _6-10 aryl group and a heteroaryl group of 5-10 atoms containing 1, 2, 3 or 4 heteroatoms independently selected from O, S or N is optionally substituted with 1, 2, 3 or 4 substituents, said The substituents are independently selected from D, F, Cl, Br, I, -CN, -NO ₂ , N ₃ , -OR ^a , -SR ^a , -NR ^a R ^b , -C(=O)NR ^a R ^b , C _1-6 alkyl, C _1-6 haloalkyl, C _2-6 alkenyl, C _2-6 alkynyl, -(C _1-4 alkylene)-CN, -(C _1-4 Alkyl)-OR ^a , -(C _1-4 alkylene)-NR ^a R ^b , C _6-10 aryl or containing 1, 2, 3 or 4 heteroatoms independently selected from O, S or N A heteroaryl group of 5-10 atoms;

In some embodiments, X is phenyl and can be optionally substituted with 1, 2, 3 or 4 independently selected from D, F, Cl, Br, C _1-3 alkyl or C _1-3 haloalkyl base replaced.

Z is C _1-6 alkyl, C _3-6 cycloalkyl, C _3-6 heterocyclyl, C _5-12 fused bicyclyl, C _5-12 spiro bicyclyl, -(C _1-4 alkylene Base)-(C _{3-6cycloalkyl} ),-(C _1-4alkylene )-(C _{3-6heterocyclyl} ),-(C _1-4alkylene )-(C _5-12 Fused bicyclyl) or -(C _1-4 alkylene)-(C _5-12 spiro bicyclyl), wherein, the C _1-6 alkyl, C _3-6 cycloalkyl, C _3-6 Heterocyclic group, C _5-12 fused bicyclic group, C _5-12 spirobicyclic group, -(C _1-4 alkylene)-(C _3-6 cycloalkyl), -(C _1-4 alkylene Base)-(C _3-6 heterocyclyl), -(C _1-4 alkylene)-(C _5-12 fused bicyclic group) and -(C _1-4 alkylene)-(C _{5- 12} spirobicyclyl), optionally substituted by 1, 2, 3 or 4 substituents independently selected from D, F, Cl, Br, I, -CN, -NO ₂ , N ₃ , C _1-4 alkyl, -OR ^a , -SR ^a , -NR ^a R ^b or -C(=O)NR ^a R ^b , and, C _5-12 fused bicyclyl, C _5-12 spirobicyclic Each ring in -(C _1-4 alkylene)-(C _5-12 fused bicyclic group) or -(C _1-4 alkylene)-(C _5-12 spiro bicyclic group) is independently is carbocyclic or heterocyclic;

In some embodiments, Z is C _1-6 alkyl, C _3-6 cycloalkyl, C _3-6 heterocyclyl, C _5-12 fused bicyclyl, C _5-12 spiro bicyclyl, -( C _1-4 alkylene)-(C _3-6 heterocyclyl), -(C _1-4 alkylene)-(C _5-12 fused bicyclic group) or -(C _1-4 alkylene )-(C _5-12 spiro bicyclyl), wherein, the C _1-6 alkyl, C _3-6 cycloalkyl, C _3-6 heterocyclyl, C _5-12 fused bicyclyl, C _{5 -12} spiro bicyclyl, -(C _1-4 alkylene)-(C _3-6 cycloalkyl),-(C _1-4 alkylene)-(C _3-6 heterocyclyl),-( C _1-4 alkylene)-(C _5-12 fused bicyclyl) and -(C _1-4 alkylene)-(C _5-12 spiro bicyclyl), optionally replaced by 1,2,3 or substituted by 4 substituents independently selected from D, F, Cl, Br, I, -CN, -NO ₂ , N ₃ , C _1-3 alkyl, -OR ^a , -NR ^a R ^b , and, C _5-12 fused bicyclyl, C _5-12 spiro bicyclyl, -(C _1-4 alkylene)-(C _5-12 fused bicyclyl) and -(C _1-4 Each ring in alkylene)-(C _5-12 spiro bicyclyl) is independently carbocyclic or heterocyclic;

Specifically, Z can be selected from any of the substructures shown in formulas (Z1) to (Z42) or their stereoisomers:

or its stereoisomers, wherein:

n is 0, 1, 2 or 3;

each W and W' is independently -O- or -N(H)-;

The substructures represented by formulas (Z1) to (Z42) or their stereoisomers are optionally 1, 2, 3 or 4 independently selected from D, F, Cl, Br, I, -CN, -NO ₂ , N ₃ , C _1-3 alkyl, -OR ^a , -NR ^a R ^b substituents, wherein, when Z is the substructure shown in formula (Z25) ~ (Z32) or its stereoisomer In the conformation, -NR ^a R ^b cannot be -NH ₂ or -NMe ₂ .

In each of the above structures of Z, each R ^a and R ^b are independently H, C _1-6 aliphatic, C _3-6 cycloalkyl, C _3-6 heterocyclyl, C _6-10 aryl, including 1, 2, 3 or 4 heteroaryl groups of 5-10 atoms independently selected from O, S or N heteroatoms, -(C _1-4 alkylene)-(C _3-6 cycloalkyl) , -(C _1-4 alkylene) -(C _3-6 heterocyclyl), -(C _1-4 alkylene) -(C _6-10 aryl) or -(C _1-4 alkylene base)-(heteroaryl group of 5-10 atoms containing 1, 2, 3 or 4 heteroatoms independently selected from O, S or N), and, when R ^a and R ^b are connected to the same nitrogen atom When, R ^a , R ^b , together with the nitrogen atom connected to them, can also optionally form a heterocyclic group with 3-8 atoms, wherein, the C _1-6 aliphatic, C _3-6 cycloalkane Base, C _3-6 heterocyclyl, C _6-10 aryl, 5-10 atom heteroaryl containing 1, 2, 3 or 4 heteroatoms independently selected from O, S or N, -( C _1-4 alkylene)-(C _3-6 cycloalkyl),-(C _1-4 alkylene)-(C _3-6 heterocyclyl),-(C _1-4 alkylene) -(C _6-10 aryl), -(C _1-4 alkylene)-(hetero Aryl) and heterocyclic group of 3-8 atoms are optionally selected from 1, 2, 3 or 4 independently selected from D, F, Cl, -CN, N ₃ , -OH, -NH ₂ , alkoxy Substituents of substituents of radicals or alkylamino groups;

In a special case, when Z is C _1-6 alkyl, -NR ^a R ^b cannot be -NH ₂ or -NMe ₂ , that is, R ^a and R ^b cannot be H or methyl at the same time.

In some embodiments, each R ^a and R ^b is independently H, C _1-3 alkyl, C _3-6 cycloalkyl or -(C _1-4 alkylene)-(C _3-6 cycloalkane group), and, when R ^a and R ^b are connected to the same nitrogen atom, R ^a , R ^b , together with the nitrogen atom connected to them, can also optionally form a heterocyclic group with 3-6 atoms, Wherein, the C _1-3 alkyl group, C _3-6 cycloalkyl group, -(C _1-4 alkylene group)-(C _3-6 cycloalkyl group) or heterocyclic group with 3-6 atoms is optional is optionally substituted with 1, 2, 3 or 4 substituents independently selected from D, F, Cl, -OH, -NH ₂ , C _1-3 alkoxy or C _1-3 alkylamino.

In a special case, when Z is C _1-6 alkyl, -NR ^a R ^b cannot be -NH ₂ or -NMe ₂ , that is, R ^a and R ^b cannot be H or methyl at the same time.

In the present invention, when R ^a and R ^b are connected to the same nitrogen atom, R ^a , R ^b and the nitrogen atom connected to them can also optionally form a heterocyclic group with 3-8 atoms or a 3- A heterocyclic group with 6 atoms, that is, R ^a , R ^b and the nitrogen atom connected to them can form a heterocyclic group with 3-8 atoms or a heterocyclic group with 3-6 atoms, or not form a heterocyclic group Cyclic group is other structures well known to those skilled in the art, such as NR ^a -R ^b or R ^a -NR ^b , etc.

The compounds of the formula (I) structure described in the present invention include but are not limited to the following specific compounds or their stereoisomers, geometric isomers, tautomers, nitrogen oxides, solvates, metabolites, pharmaceutically available Accepted salts or its prodrugs:

Unless otherwise indicated, all stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, metabolites, salts and pharmaceutically acceptable prodrugs of the compounds of the present invention are Belong to the scope of the present invention.

In particular, the salts are pharmaceutically acceptable salts. The term "pharmaceutically acceptable" includes that the substance or composition must be chemically or toxicologically appropriate in relation to the other ingredients making up the formulation and the mammal being used for treatment.

The salts of the compounds of the present invention also include the salts of the intermediates of the compounds shown in the formula (I) or the separated enantiomers used in the preparation or purification of the compounds shown in the formula (I), but not necessarily pharmaceutically acceptable Salt.

If the compound of the present invention is basic, the desired salt may be prepared by any suitable method provided in the literature, for example, using inorganic acids such as hydrochloric, hydrobromic, sulfuric, nitric and phosphoric acids and the like. Alternatively use organic acids such as acetic, maleic, succinic, mandelic, fumaric, malonic, pyruvic, oxalic, glycolic and salicylic acids; pyranonic acids such as glucuronic and galactose Alkyd acids; α-hydroxy acids such as citric acid and tartaric acid; amino acids such as aspartic acid and glutamic acid; aromatic acids such as benzoic acid and cinnamic acid; sulfonic acids such as p-toluenesulfonic acid, ethanesulfonic acid, etc.

If the compound of the invention is acidic, the desired salts can be prepared by suitable methods, e.g., using inorganic or organic bases, such as ammonia (primary, secondary, tertiary), alkali metal hydroxides or alkaline earth metal hydroxides, etc. Suitable salts include, but are not limited to, organic salts derived from amino acids such as glycine and arginine, ammonia such as primary, secondary and tertiary ammonia, and cyclic ammonia such as piperidine, morpholine and piperazine etc., and inorganic salts obtained from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum and lithium.

The present invention also includes the use of the compounds of the present invention and pharmaceutically acceptable salts thereof, that is, for the production of pharmaceutical products for the treatment of acute and chronic angiogenesis-mediated diseases, including those described in the present invention in the production of anticancer drugs . The compounds of the present invention are also useful in the manufacture of a medicinal product for alleviating, preventing, controlling or treating diseases mediated by ALK or c-Met. The present invention includes a pharmaceutical composition, which comprises a compound represented by formula (I) in combination with at least one pharmaceutically acceptable carrier, adjuvant or diluent in a therapeutically effective amount.

The present invention also includes a method of treating an angiogenesis-mediated disease in a patient, or being susceptible to it, comprising treating the patient with a therapeutically effective amount of a compound represented by formula (I).

combination

According to another aspect, the characteristics of the pharmaceutical composition of the present invention include the compound of formula (I), the compound listed in the present invention, or the compound in the examples, and a pharmaceutically acceptable carrier, adjuvant, or vehicle agent. The amount of compound in the compositions of the invention is effective to detectably inhibit a protein kinase in a biological specimen or patient.

The compounds of the present invention exist in free form, or suitably, as pharmaceutically acceptable derivatives. According to the present invention, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable prodrugs, salts, esters, salts of esters, or any other compounds that can be administered directly or indirectly according to the needs of patients. Adducts or derivatives, compounds described in other aspects of the present invention, their metabolites or their residues.

As described herein, the pharmaceutically acceptable composition of the present invention further comprises a pharmaceutically acceptable carrier, adjuvant, or vehicle, which, as used in the present invention, includes any solvent, diluent, or other Liquid excipients, dispersing or suspending agents, surfactants, isotonic agents, thickening agents, emulsifying agents, preservatives, solid binders or lubricants, etc., are suitable for the specific target dosage form. As described in: In Remington: The Science and Practice of Pharmacy, 21st edition, 2005, ed.D.B.Troy, LippincottWilliams&Wilkins, Philadelphia, and Encyclopedia of Pharmaceutical Technology, eds.J.Swarbrick and J.C.Marcel, 1988-1999k , New York, summarizing the contents of the literature herein, shows that different carriers can be used in the formulation of pharmaceutically acceptable compositions and their known methods of preparation. Except to the extent that any conventional carrier media is incompatible with the compounds of the present invention, such as any adverse biological effects produced or interactions in a deleterious manner with any other components of the pharmaceutically acceptable composition, they The purposes of the present invention are also considered scope.

Substances that can be used as pharmaceutically acceptable carriers include, but are not limited to, ion exchangers, aluminum, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphate, glycine, sorbic acid, sorbic acid, Potassium phosphate, partial glyceride mixture of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silicon, magnesium trisilicate, polyethylene Pyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene-blocking polymers, lanolin, sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as carboxymethyl sodium cellulose, ethyl cellulose, and cellulose acetate; gum powder; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, Olive, corn, and soybean oils; glycols, such as propylene glycol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; seaweed acid; pyrogen-free water; isotonic salts; Ringer's solution; ethanol, phosphate buffered solution, and other nontoxic suitable lubricants such as sodium lauryl sulfate and magnesium stearate, colorants, release agents, Coatings, sweeteners, flavors and fragrances, preservatives and antioxidants.

The compositions of the present invention may be administered orally, by injection, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implantable kit . The term "injected" as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial (cavity), intrasternal, intrathecal, intraocular, intrahepatic, focal Intracranial, and intracranial injection or infusion techniques. Preferred compositions are administered orally, intraperitoneally or intravenously. Sterile injectable forms of the compositions of the present invention may be aqueous or oleaginous suspensions. These suspensions may be formulated according to the known art using suitable dispersing agents, wetting agents and suspending agents. Sterile injections can be sterile injection solutions or suspensions, which are non-toxic acceptable diluents or solvents for injection, such as 1,3-butanediol solution. Among the acceptable vehicles and solvents are water, Ringer's solution and isotonic sodium chloride solution. Furthermore, sterile, fixed oils are conventionally employed as a solvent or suspending medium.

For this purpose any bland fixed oil may be a synthetic mono- or diglyceride. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially their polyoxyethylene derivatives. These oil solutions or suspensions may contain a long-chain alcohol diluent or dispersant, such as carboxymethylcellulose or similar dispersing agents, commonly used in pharmaceutical formulations of pharmaceutically acceptable dosage forms including emulsions and suspensions. Other commonly used surfactants, such as Tweens, Spans and other emulsifiers or enhancers of bioavailability, are generally used in pharmaceutically acceptable solid, liquid, or other dosage forms, and can be applied to target pharmaceutical preparations preparation.

The pharmaceutically acceptable compositions of this invention may be orally administered in any acceptable oral dosage form including, but not limited to, capsules, tablets, aqueous suspensions or solutions. For tablets for oral use, carriers generally include lactose and corn starch. Lubricants, such as magnesium stearate, are typically added. For oral administration in capsules, suitable diluents include lactose and dried cornstarch. When administered orally as an aqueous suspension, the active ingredient consists of emulsifying and suspending agents. Certain sweetening, flavoring or coloring agents may also be added if desired in these dosage forms.

Additionally, the pharmaceutically acceptable compositions of this invention may be administered rectally in the form of suppositories. These can be prepared by mixing the agent with a suitable non-infusing excipient which is solid at room temperature but liquid at rectal temperature, where it melts and releases the drug. Such materials include cocoa butter, beeswax, and polyethylene glycols. The pharmaceutically acceptable compositions of the present invention may be administered topically, especially when topical administration involves areas or organs where the therapeutic target is easily accessible, such as diseases of the eyes, skin or lower intestinal tract. Suitable topical formulations can be prepared and applied to these areas or organs.

The aforementioned rectal suppositories or suitable enemas may be applied topically to the lower intestinal tract. Local skin spots can also be used in this way. For topical use, the pharmaceutically acceptable compositions may be formulated in a suitable ointment containing the active ingredient suspended or dissolved in one or more carriers. Carrier compounds for topical administration of this invention include, but are not limited to, mineral oil, liquid paraffin, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water. Alternatively, the pharmaceutically acceptable compositions may be prepared in a suitable lotion or cream containing the active ingredients suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, Span-60 (sorbitan monostearate), Tween 60 (polysorbate 60), cetyl esters wax, palmityl alcohol, 2- Octyldodecanol, Benzyl Alcohol and Water.

For ophthalmic use, the pharmaceutically acceptable composition can be prepared as a formulation, such as isotonic micronized suspension, pH-adjusted sterile saline or other aqueous solution, preferably, isotonic solution and pH-adjusted sterile saline or For other aqueous solutions, disinfectant preservatives such as benzalkonium chloride can be added. In addition, for ophthalmic use, the pharmaceutically acceptable composition can be formulated into an ointment such as petrolatum. The pharmaceutically acceptable compositions of this invention may be administered by nasal aerosol or inhalation. Such compositions may be prepared according to known techniques of formulation formulation, or may be prepared as saline solutions using benzyl alcohol or other suitable preservatives, absorption enhancers, fluorocarbons or other conventional solubilizing or dispersing agents to enhance bioavailability Spend.

Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. Liquid dosage forms may contain, in addition to the active compound, generally known inert diluents, for example, water or other solvents, solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzoic acid Benzyl esters, propylene glycol, 1,3-butanediol, dimethylformamide, oils and fats (especially cottonseed, groundnut, corn, microbial, olive, castor and sesame oils), glycerin, 2-tetrahydrofurfurylmethanol, polyethylene glycol , sorbitan fatty acid esters, and mixtures thereof. Besides inert diluents, the oral compositions can also contain adjuvants such as wetting agents, emulsifying or suspending agents, sweetening, flavoring, and perfuming agents.

Injections, such as sterile injections or oily suspensions, can be prepared according to known techniques using suitable dispersing agents, wetting agents and suspending agents according to formulations. Sterile injectable preparations can be sterile injectable solutions, suspensions or emulsions prepared with non-toxic injectable acceptable diluents or solvents, for example, 1,3-butanediol solution. Among the acceptable vehicles and solvents are water, Ringer's solution, U.S.P. and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. Any bland fixed oil may be used for this purpose including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in injectables.

The injection can be sterile, such as filtered through a bacteria-defense filter, or incorporated in the form of a sterile solid composition with a sterilizing agent that can be dissolved or dispersed in sterile water or other sterile injectable media before use middle. In order to prolong the effect of the compounds of the invention, it is usually necessary to slow the absorption of the compounds by subcutaneous or intramuscular injection. In this way, the liquid suspension can be used to solve the problem of poor water solubility of crystalline or amorphous substances. The rate of absorption of a compound depends upon its rate of dissolution, which in turn depends on crystal size and crystalline shape. Additionally, delayed absorption of the compound administered by injection can be accomplished by dissolving or dispersing the compound in an oil vehicle.

Injectable depot forms are made by forming microencapsule matrices of the compound in biodegradable polymers, such as polylactide-polyglycolide. The rate at which the compound is released depends on the rate at which the compound forms the polymer and the nature of the particular polymer. Other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Injectable depot forms are also prepared by entrapping the compound in liposomes or microemulsions which are compatible with body tissue.

In some of these embodiments, the composition for rectal or vaginal administration is a suppository, which can be prepared by mixing the compound of the present invention with a suitable non-infusion adjuvant or carrier, such as cocoa butter, polyethylene glycol, or Suppositories are waxy substances that are solid at room temperature but liquid at body temperature and therefore melt in the vaginal or thecal cavity to release the active compound.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these dosage forms, the active compound is mixed with at least one pharmaceutically acceptable inert excipient or carrier, such as sodium citrate or calcium phosphate or fillers or a) fillers such as starch, lactose, sucrose, glucose, mannitol and Silicic acid, b) binders such as carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and gum arabic, c) humectants such as glycerin, d) disintegrants such as agar, calcium carbonate, potatoes Starch or tapioca starch, alginic acid, certain silicates and sodium carbonate, e) blocker solutions such as paraffin, f) absorption enhancers such as quaternary ammonium compounds, g) humectants such as cetyl alcohol and glycerol monostearate Esters, h) absorbents such as kaolin and bentonite, i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof. As with capsules, tablets and pills, these dosage forms may contain buffering agents.

A similar type of solid composition can be a soft or hard capsule filled with fillers, and the used excipients include lactose and macromolecular polyethylene glycol and the like. Solid dosage forms like tablets, lozenges, capsules, pills and granules can be prepared by coating, shelling such as enteric coating and other known coating methods for pharmaceutical preparations. They may optionally contain opacifying agents or, preferably, release the only active ingredient in the composition in a certain part of the intestinal tract, optionally in a delayed manner. For example implant compositions may comprise polymeric substances and waxes.

The active compounds can be in microencapsulated dosage form with one or more excipients as described herein. Solid dosage forms like tablets, lozenges, capsules, pills and granules can be coated or shelled, such as enteric coatings, release-controlling coatings and other well-known pharmaceutical formulation methods. In these solid dosage forms, the active compound may be admixed with at least one inert diluent, such as sucrose, lactose or starch. Such dosage forms may also contain, as usual, additional substances other than inert diluents, such as tableting lubricants and other tableting aids such as magnesium stearate and microcrystalline cellulose. As with capsules, tablets and pills, these dosage forms may contain buffering agents. They may optionally contain a sedative or, preferably, release the only active ingredient of the composition in a certain part of the intestinal tract, with any delay. Applicable implant compositions may include, but are not limited to, polymers and waxes.

Dosage forms for topical or dermal administration of a compound of this invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, patches. The active ingredient is mixed under sterile conditions with a pharmaceutically acceptable carrier and any necessary preservatives or necessary buffers. Ophthalmic pharmaceutical formulations, ear drops and eye drops are all contemplated by the present invention. In addition, the present invention also contemplates the application of transdermal patches, which have more advantages in controlling the delivery of the compound into the body. Such dosage forms can be prepared by dissolving or dispersing the compound in a suitable medium. Absorption enhancers can increase the flux of the compound across the skin, controlling the rate either by rate controlling films or by dispersing the compound in a polymer matrix or gelatin.

The compounds of the present invention are preferably formulated in dosage unit form to ease administration and uniformity of dosage. The term "dosage unit form" as used herein refers to a physically discrete unit of drug required to adequately treat a patient. It is to be understood, however, that the total daily usage of the compounds or compositions of the present invention will be determined by the attending physician based on sound medically sound judgment. The particular effective dosage level for any particular patient or organism will depend on many factors including the condition being treated and the severity of the condition, the activity of the particular compound, the particular composition employed, the age, weight, health, sex of the patient and dietary habits, timing of administration, route of administration and rate of excretion of the particular compound employed, duration of treatment, use of the drug in combination or with specific compounds, and other factors well known in the art of pharmacy.

The amount of a compound of the invention which may be combined with a carrier material to produce a single dosage composition will vary depending upon the indication and the particular mode of administration. In some embodiments, the composition can be prepared according to the preparation method into an inhibitor with a dosage of 0.01-200 mg/kg body weight/day, and the administration is carried out according to the amount of the composition accepted by the patient.

The compounds of the present invention may be administered as the sole pharmaceutical agent or in combination with one or more other additional therapeutic (pharmaceutical) agents, where the combination causes acceptable adverse effects, which is of interest in the treatment of hyperproliferative diseases such as cancer special meaning. In such cases, the compounds of the invention may be combined with known cytotoxic agents, individual transduction inhibitors or other anticancer agents, as well as mixtures and combinations thereof. As used herein, an additional therapeutic agent normally administered to treat a particular disease is known as "appropriately treating a disease". "Additional therapeutic agents" as used in the present invention include chemotherapeutic drugs or other anti-proliferative drugs that can be combined with the compounds of the present invention to treat proliferative diseases or cancer.

Chemotherapeutic or other antiproliferative agents including histone deacetylase (HDAC) inhibitors, including but not limited to, SAHA, MS-275, MGO103, and those compounds described in WO 2006/010264, WO 03/024448, WO2004/069823, US 2006/0058298, US 2005/0288282, WO 00/71703, WO01/38322, WO 01/70675, WO 03/006652, WO 2004/035525, WO2005/0392005/009, WO , and demethylating agents include, but are not limited to, 5-aza-2′-deoxycytidine (5-aza-dC), azacitidine (Vidaza), decitabine (Decitabine), and the following Compounds described in literature: US 6,268137, US 5,578,716, US 5,919,772, US 6,054,439, US 6,184,211, US 6,020,318, US 6,066,625, US 6,506,735, US 6,221,849, US 6,953,3183, US 6,953,3183, US 9.

In other embodiments, chemotherapeutic drugs or other anti-proliferative drugs can be combined with the compounds of the present invention to treat proliferative diseases and cancers. Known chemotherapeutic agents include, but are not limited to, other therapies or anticancer drugs that may be used in combination with the anticancer agents of the present invention, surgery, radiation therapy (a few examples are gamma radiation, neutron beam radiation therapy, electron beam radiation therapy , proton therapy, brachytherapy and systemic radioisotope therapy), endocrine therapy, taxanes (paclitaxel (Taxol), docetaxel (Taxotere) etc.), platinum derivatives (cisplatin (Cisplatin), carboplatin ( Carboplatin), oxaliplatin, satraplatin), biological response modifiers (interferon, interleukin), tumor necrosis factor (TNF, TRAIL receptor target), hyperthermia and cryotherapy, Agents (eg, antiemetics) to mitigate any adverse effects, and other approved chemotherapeutic agents, including, but not limited to, alkylating agents (mechlorethamine, chlorambucil, cyclophosphamide ( cyclophosphamide), melphalan, ifosfamide), antimetabolites (Methotrexate, Pemetrexed, etc.), purine antagonists and pyrimidine antagonists ( 6-Mercaptopurine, 5-Fluorouracil, Cytarabine, Gemcitabine), spindle inhibitors (Vinblastine, Vincristine) , vinorelbine), podophyllotoxins (etoposide, irinotecan, topotecan), antibiotics (doxorubicin, bleomycin ( Bleomycin), Mitomycin), nitrosoureas (Carmustine, Lomustine), cell division cycle inhibitors (KSP through mitotic kinesin inhibitors, CENP- E and CDK inhibitors), enzymes (Asparaginase), hormones (Tamoxifen, Leuprolide, Flutamide, Megestrol , Dexamethasone, etc.). Anti-angiogenic agents (Avastin, etc.). Monoclonal antibodies (Belimumab, Brentuximab, Cetuximab, Gemtuzumab, Ipilimumab, Ofatumumab, Panitumumab, Ranibizumab Monoclonal antibody (Ranibizumab), Rituximab (Rituximab), Tositumomab (Tositumomab), Trastuzumab (Trastuzumab)). Kinase inhibitors (Imatinib, Sunitinib, Sorafenib, Erlotinib, Gefitinib, Dasatinib ), Nilotinib, Lapatinib, Crizotinib, Ruxolitinib, Vemurafenib, Vandetanib, Pazopanib, etc.). Drugs inhibit or activate cancer pathways such as mTOR, HIF (hypoxia-inducible factor) pathway and others. See http://www.nci.nih.gov/ for a broader forum on cancer treatment, http://www.fda.gov/cder/cancer/druglist-rame.htm for a list of FAD-approved oncology drugs, and Gram Handbook, Eighteenth Edition. 2006, all contents are incorporated by reference.

In other embodiments, the compounds of the present invention can be combined with cytotoxic anticancer agents. Such anticancer agents can be found in the Thirteenth Edition of The Merck Index (2001). These anticancer agents include, but are by no means limited to, asparaginase, bleomycin, carboplatin, carmustine, chlorambucil, cisplatin, L-asparaginase, cyclophosphamide, albino Cytosine, dacarbazine, actinomycin D, daunorubicin, doxorubicin (doxorubicin), epirubicin, etoposide, 5-fluorouracil, hexamethylmelamine, hydroxy Urea, ifosfamide, irinotecan, leucovorin, cyclohexylnitrosourea, nitrogen mustard, 6-mercaptopurine, mesna, methotrexate, mitomycin C, mitoxantrone, prednisolone Thiolone, prednisone, procarbazine, raloxifene, streptozotocin, tamoxifen, thioguanine, topotecan, vinblastine, vincristine, and vindesine.

Other suitable cytotoxic drugs for use in combination with the compounds of the present invention include, but are not limited to, those compounds that are recognized for use in the treatment of neoplastic diseases, as described in Goodman and Gilman's The Pharmacological Basis of Therapeutics (Ninth Edition, 1996, McGraw-Hill.); these anticancer agents include, but are not limited to, aminoglutethimide, L-asparaginase, azathioprine, 5-azacytidine , Cladribine, Busulfan, Diethylstilbestrol, 2,2′-difluorodeoxyciticoline, Docetaxel, Erythrohydroxynonyladenine, Ethinylestradiol Alcohol, 5-fluorouridine deoxynucleoside, 5-fluorodeoxyuridine monophosphate, fludarabine phosphate, fluoxymesterone, flutamide, hydroxyprogesterone caproate, ida Idarubicin, Interferon, Medroxyprogesterone Acetate, Megestrol Acetate, Melphalan, Mitotane, Paclitaxel, Pentostatin, N-Phosphoacetyl -L-Aspartic Acid (PALA), Plicamycin (Plicamycin), Methylcyclohexylnitrosourea (Semustine), Teniposide (Teniposide), Testosterone Propionate, Thiotepa, Tri Methylmelamine, uridine, and vinorelbine.

Other cytotoxic anticancer agents suitable for use in combination with the compounds of the present invention include newly discovered cytotoxic substances, including, but not limited to, oxaliplatin (Oxaliplatin), gemcitabine (Gemcitabine), capecitabine (Capecitabine), a macrolide antineoplastic drug and its natural or synthetic derivatives, Temozolomide (Quinn et al., J. Clin. Oncology, 2003, 21 (4), 646-651), Torto Siblomab (Bexxar), Trabedectin (Vidal et al., Proceedings of the American Society for Clinical Oncology, 2004, 23, abstract 3181), and the kinesin spindle protein inhibitor Eg5 (Wood et al., Curr. Opin. Pharmacol. 2001, 1, 370-377).

In other embodiments, the compounds of the invention may be combined with other signal transduction inhibitors. Interestingly, signal transduction inhibitors target the EGFR family, such as EGFR, HER-2 and HER-4 (Raymond et al., Drugs, 2000, 60(Suppl.l), 15-23; Harari et al., Oncogene, 2000, 19(53), 6102-6114) and their respective ligands. Such agents include, but are by no means limited to, antibody therapies such as Trastuzumab, Cetuximab, Ipilimumab and Pertuzumab. Such therapies also include, but are by no means limited to, small molecule kinase inhibitors such as Imatinib, Sunitinib, Sorafenib, Erlotinib, Gem Gefitinib, Dasatinib, Nilotinib, Lapatinib, Crizotinib, Ruxolitinib, Vemurafenib, Vandetanib, Pazopanib, Afatinib (Afatinib), amuvatinib, axitinib, bosutinib, brivanib, canertinib, cabozantinib, cediranib, dabrafenib, dacomitinib, danusertib, dovitinib, foretinib, ganetespib, ibrutinib , iniparib, lenvatinib, linifanib, linsitinib, masitinib, momelotinib, motesanib, neratinib, niraparib, oprozomib, olaparib, pictilisib, ponatinib, quizartinib, regorafenib, rigosertib, Rucaparib, saracatinib, saridegib, tandutinib, tasocitinib, telatinib, tivantinib, tivozanib, tofacitinib, trametinib, vatalanib, veliparib, vismodegib, volasertib, BMS-540215, BMS777607, JNJ38877605, TKI258, GDC-0941 ( et al., J.Med.Chem.2008, 51, 5522), BZE235, etc.

In other embodiments, the compounds of the invention can bind to histone deacetylase inhibitors. Such reagents include, but are by no means limited to, suberoylanilide hydroxamic acid (SAHA), LAQ-824 (Ottmann et al., Proceedings of the American Society for Clinical Oncology, 2004, 23, abstract 3024), LBH-589 (Beck et al., Proceedings of the American Society for Clinical Oncology, 2004, 23, abstract 3025), MS-275 (Ryan et al., Proceedings of the American Association of Cancer Research, 2004, 45, abstract 2452), FR-901228 (Piekarz et al. ., Proceedings of the American Society for Clinical Oncology, 2004, 23, abstract 3028) and MGCDOI03 (US6,897,220).

In other embodiments, the compounds of the invention may be combined with other anticancer agents such as proteasome inhibitors and m-TOR inhibitors. These include, but are by no means limited to, Bortezomib (Mackay et al., Proceedings of the American Society for Clinical Oncology, 2004, 23, Abstract 3109), and CCI-779 (Wu et al., Proceedings of the American Association of Cancer Research, 2004, 45, abstract 3849). The compounds of the invention may also be combined with other anticancer agents such as topoisomerase inhibitors, including but by no means limited to camptothecin.

Those additional therapeutic agents may be administered separately from the composition comprising the compound of the invention as part of a multiple dosing regimen. Alternatively, those therapeutic agents may be part of a single dosage form, mixed together with the compounds of this invention to form a single composition. If administered as part of a multiple dosing regimen, the two active agents may be delivered to each other simultaneously, sequentially or over a period of time, such that the desired agent activity is achieved.

The amount of the compound and the additional therapeutic agent (those compositions containing an additional therapeutic agent such as those described herein) which can be combined to produce a single dosage form will vary depending on the indication and the particular mode of administration. Normally, the amount of additional therapeutic agent in the compositions of the invention will not exceed the amount normally administered for a composition comprising the therapeutic agent as the only active agent. In another aspect, the disclosed compositions additional therapeutic agent in an amount ranging from about 50% to 100% of the normal amount of prior compositions, comprising the agent as the sole active therapeutic agent. In those compositions comprising an additional therapeutic agent, the additional therapeutic agent will act synergistically with the compound of the invention.

Application of Compounds and Compositions

The above-mentioned compounds and pharmaceutical compositions provided by the present invention can be used to prepare medicines for preventing, treating, treating or alleviating proliferative diseases, and can also be used for preparing medicines for inhibiting or regulating protein kinase activity.

In particular, the amount of compound in the compositions of the invention is effective to detectably inhibit the activity of a protein kinase such as ALK or c-Met. The compounds of the present invention will be used as antineoplastic agents to treat patients or reduce the deleterious effects of ALK and c-Met signaling responses.

The compounds of the present invention may be used in, but by no means limited to, administering to a patient an effective amount of the compound or composition of the present invention to prevent or treat a proliferative disease in a patient. Such diseases include cancer, especially metastatic cancer, atherosclerosis and pulmonary fibrosis.

The compounds of the present invention can be applied to the treatment of tumors, including cancer and metastatic cancer, further including but not limited to, cancers such as bladder cancer, breast cancer, colon cancer, kidney cancer, liver cancer, lung cancer (including small cell lung cancer), esophageal cancer , gallbladder, ovary, pancreas, stomach, cervix, thyroid, prostate, and skin (including squamous cell carcinoma); hematopoietic neoplasms of the lymphatic system (including leukemia, acute lymphocystic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell leukemia and Burkitt's lymphoma); hematopoietic tumors of the myeloid system (including acute chronic myelogenous myelogenous leukemia, myelodysplastic syndrome, and promyelocytic leukemia); tumors of mesenchymal origin (including fibrosarcoma and rhabdomyosarcoma, and other sarcomas, such as soft tissue and cartilage); central peripheral nervous system tumors (including astrocytoma, neuroblastoma, glioma, and schwannoma); and other tumors (including melanoma, seminoma, teratocarcinoma, osteosarcoma, xenoderoma pigmentosum, keratoctanthoma, thyroid Follicular tumor and Kaposi's sarcoma).

The compounds and pharmaceutical compositions of the present invention are also useful in the treatment of ophthalmic conditions such as corneal transplant rejection, neovascularization of the eye, retinal neovascularization including neovascularization following injury or infection; diabetic retinopathy; retrolentic fibroplasia , and neovascular glaucoma; retinal ischemia; vitreous hemorrhage; ulcerative diseases such as gastric ulcer; tumors; disorders of the female reproductive system such as endometriosis. These compounds are also useful in the treatment of edema and vascular hyperpermeability conditions.

The compounds and pharmaceutical compositions of the present invention are also useful in the management of conditions associated with diabetes such as diabetic retinopathy and microangiopathy. The compounds of the invention are also useful in cases of reduced blood flow in cancer patients. The compounds of the invention also have a beneficial effect on the reduction of tumor metastasis in patients.

In addition to being useful in human therapy, the compounds and pharmaceutical compositions of the present invention can also be used in veterinary treatment of pets, imported animals and farm animals, including mammals, rodents and the like. Examples of additional animals include horses, dogs and cats. Here, the compounds of the present invention include their pharmaceutically acceptable derivatives.

Where the plural form is applied to a compound, salt, etc., it also means a single compound, salt, etc.

The method of treatment comprising the administration of the compound or composition of the present invention further comprises the administration of an additional therapeutic agent (combination therapy) to the patient, wherein the additional therapeutic agent is selected from: chemotherapy, anti-proliferative or anti-inflammatory agents, wherein the additional therapeutic The agent is appropriate for the condition being treated, and the additional therapeutic agent may be administered in combination with the compound or composition of the invention as a single dosage form, or as separate compounds or compositions as part of a multiple dosage form. Additional therapeutic agents may be administered at the same time or at different times as the compounds of the invention. In the latter case, the administration may be staggered for 6 hours, 12 hours, 1 day, 2 days, 3 days, 1 week, 2 weeks, 3 weeks, 1 month or 2 months.

The present invention also includes a method of inhibiting the growth of cells expressing ALK or c-Met, the method comprising contacting the cell with a compound or composition of the present invention, thereby inhibiting the growth of the cell. Cells whose growth can be inhibited include: breast cancer cells, colorectal cancer cells, lung cancer cells, papillary cancer cells, prostate cancer cells, lymphoma cells, colon cancer cells, pancreatic cancer cells, ovarian cancer cells, cervical cancer cells, Central nervous system cancer cells, osteosarcoma cells, kidney cancer cells, hepatocellular carcinoma cells, bladder cancer cells, gastric cancer cells, head or neck squamous cell carcinoma cells, melanoma cells and leukemia cells.

The invention provides a method of inhibiting ALK or c-Met kinase activity in a biological sample, the method comprising contacting a compound or composition of the invention with the biological sample. The term "biological specimen" used in the present invention refers to specimens outside the living body, including but not limited to, cell culture or cell extraction; biopsy material obtained from mammals or their extracts; blood, saliva, urine, Feces, semen, tears, or other liquid substances of living tissue and their extracts. Inhibition of kinase activity, particularly ALK or c-Met kinase activity, in a biological specimen can be used in a variety of ways well known to those skilled in the art. Such uses include, but are by no means limited to, blood transfusions, organ transplantation, biospecimen storage, and bioassays.

An "effective amount" or "effective dose" of a compound or pharmaceutically acceptable composition of the present invention refers to an effective amount for treating or reducing the severity of one or more of the conditions mentioned in the present invention. According to the methods of the present invention, the compounds and compositions may be administered in any amount and by any route of administration effective for treating or lessening the severity of a disease. The exact amount necessary will vary from patient to patient, depending on race, age, general condition of the patient, severity of infection, particular factors, mode of administration, and the like. A compound or composition may be administered in combination with one or more other therapeutic agents, as discussed herein.

The compounds of the present invention or their pharmaceutical compositions can be applied to the coating of implantable medical devices, such as prostheses, artificial valves, artificial blood vessels, stems and urinary catheters. Vascular stems, for example, have been used to overcome restenosis (reconstriction of vessel walls after injury). However, patients using stems or other implantable devices will be at risk of clot formation or platelet activation. These adverse effects can be prevented or mitigated by pre-coating the device with a pharmaceutically acceptable composition comprising a compound of the invention.

Suitable coatings and general preparation of coatings for implantable devices are described in documents US 6,099,562; US 5,886,026; and US 5,304,121, the coatings being typically biocompatible polymeric materials such as hydrogel Gum polymer, polymethyldisiloxane, polycaprolactone, polyethylene glycol, polylactic acid, ethylene vinyl acetate, and mixtures thereof. The coating may optionally be further covered with a suitable coating, such as fluorosimethicone, polysaccharase, polyethylene glycol, phospholipids, or a combination thereof, to exhibit controlled-release characteristics of the composition. Another aspect of the invention includes implantable devices coated with a compound of the invention. The compounds of the present invention may also be coated on implantable medical devices, such as beads, or mixed with polymers or other molecules to provide "drug depots", thus allowing drug Release has a longer time period.

Compound Synthesis Method

In order to describe the present invention, examples are listed below. However, it should be understood that the present invention is not limited to these examples, but only provides a method of practicing the present invention.

Generally, the compounds of the present invention can be prepared by the methods described in the present invention, and unless otherwise specified, the definitions of the substituents are as shown in formula (I). The following reaction schemes and examples serve to further illustrate the present invention.

Those skilled in the art will recognize that the chemical reactions described herein can be used to suitably prepare many other compounds of the invention and that other methods for preparing the compounds of the invention are considered to be within the scope of the invention Inside. For example, the synthesis of those non-exemplified compounds according to the present invention can be successfully accomplished by those skilled in the art through modification methods, such as appropriate protection of interfering groups, by using other known reagents in addition to those described in the present invention, or by incorporating Reaction conditions with some routine modifications. In addition, reactions disclosed herein or known reaction conditions are also recognized to be applicable to the preparation of other compounds of this invention.

In the examples described below, unless indicated otherwise, all temperatures are in degrees Celsius. Reagents were purchased from commercial suppliers such as Aldrich Chemical Company, Arco Chemical Company and Alfa Chemical Company and used without further purification unless otherwise indicated. General reagents from Shantou Xilong Chemical Factory, Guangdong Guanghua Chemical Reagent Factory, Guangzhou Chemical Reagent Factory, Tianjin Haoyuyu Chemical Co., Ltd., Tianjin Fuchen Chemical Reagent Factory, Wuhan Xinhuayuan Technology Development Co., Ltd., Qingdao Tenglong Chemical Reagent Co., Ltd., and Qingdao Ocean Chemical Factory can be purchased.

Anhydrous tetrahydrofuran, dioxane, toluene, and ether were obtained by reflux drying over sodium metal. Anhydrous dichloromethane and chloroform were obtained by refluxing and drying over calcium hydride. Ethyl acetate, petroleum ether, n-hexane, N,N-dimethylacetamide and N,N-dimethylformamide were dried over anhydrous sodium sulfate before use.

The following reactions are generally carried out under positive pressure of nitrogen or argon or over anhydrous solvents with a drying tube (unless otherwise indicated), the reaction vials are all stoppered with suitable rubber stoppers, and the substrate is injected by syringe. Glassware is dried.

The chromatographic column is a silica gel column. Silica gel (300-400 mesh) was purchased from Qingdao Ocean Chemical Factory. The nuclear magnetic resonance spectrum uses CDC1 ₃ , d ₆ -DMSO, CD ₃ OD or d ₆ -acetone as the solvent (reported in ppm), and TMS (0ppm) or chloroform (7.25ppm) as the reference standard. When multiplets appear, the following abbreviations will be used: s (singlet, singlet), d (doublet, doublet), t (triplet, triplet), m (multiplet, multiplet), br (broadened, broadened) peak), dd (doublet of doublets, quartet), dt (doublet of triplets, double triplet). Coupling constants are expressed in Hertz (Hz).

The conditions for low-resolution mass spectrometry (MS) data are: Agilent1200 or Agilent6120Series LCMS (column model: Zorbax SB-C18, 2.1×30mm, 3.5 microns, 6min, flow rate 0.6mL/min. Mobile phase: 5-95% (containing 0.1% formic acid (CH ₃ CN) in (containing 0.1% formic acid in H ₂ O), using UV detection at 210/254nm, using low response electrospray mode (ESI); Agilent7890A-5975C Series GCMS (column model: DB-624, 30mm×0.32mm, 1.8 micron), single quadrupole mass analyzer.

Pure compounds were characterized by Agilent 1100 Series High Performance Liquid Chromatography (HPLC) with UV detection at 210 nm and 254 nm. Columns are typically operated at 40°C.

The following abbreviations are used throughout this disclosure:

BBr ₃ boron tribromide

BINAP 2,2′-bisdiphenylphosphino-1,1′-binaphthyl

BOC, Boc tert-butoxycarbonyl

BSA bovine serum albumin

CDC1 ₃ deuterated chloroform

CHCl ₃ Chloroform

_CH2Cl2 , DCM _{dichloromethane}

CH ₃ SO ₂ Cl, MsCl p-toluenesulfonyl chloride

Cs ₂ CO ₃ cesium carbonate

Cu Copper

CuI Cuprous iodide

DAST diethylaminosulfur trifluoride

DBU 1,8-Diazabicyclo[5.4.0]undec-7-ene

DEAD diethyl azodicarboxylate

DEAD diethyl azodicarboxylate

DIAD Diisopropyl azodicarboxylate

DIBAL diisobutylaluminum hydride

DIEA, DIPEA Diisopropylethylamine

DMAP 4-Dimethylaminopyridine

DMF N,N-Dimethylformamide

DMSO dimethyl sulfoxide

DPPA diphenylphosphoryl azide

EDCI 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride

EtOAc, EA ethyl acetate

Et ₂ O diethyl ether

Et ₃ N, TEA Triethylamine

FBS fetal bovine serum

Fe iron

g grams

h hours

HATU O-(7-Azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate

HBr hydrobromic acid

HBTU O-benzotriazole-N,N,N′,N′-tetramethyluronium hexafluorophosphate

HCl hydrochloric acid

H ₂ hydrogen

_H2O water

H ₂ O ₂ hydrogen peroxide

HOAc, AcOH acetic acid

HOBt 1-Hydroxybenzotriazole

_{K2CO3potassium carbonate}

KOH potassium hydroxide

LiHMDS lithium hexamethyldisilazide

LDA lithium diisopropylamide

MCPBA m-chloroperoxybenzoic acid

MeCN, _CH3CN acetonitrile

MeI methyl iodide

MeOH, _CH3OH Methanol

2-MeTHF 2-Methyltetrahydrofuran

MgSO ₄ magnesium sulfate

mL, ml milliliter

N ₂ Nitrogen

NaBH ₄ sodium borohydride

NaBH ₃ CN Sodium cyanoborohydride

NaCl Sodium Chloride

NaClO ₂ sodium chlorite

NaH Sodium hydride

NaHCO ₃ sodium bicarbonate

NaH ₂ PO ₄ sodium dihydrogen phosphate

NaI Sodium iodide

NaO(t-Bu) sodium tert-butoxide

NaOH sodium hydroxide

Sodium Na ₂ SO ₄ Sulfate

NBS N-Bromosuccinimide

NH ₃ ammonia

NH ₄ C1 ammonium chloride

NMP N-Methylpyrrolidone

PBS Phosphate Buffered Saline

P(t-Bu) ₃ tri(tert-butyl)phosphine

Pd/C palladium/carbon

Pd ₂ (dba) ₃ Bis(dibenzylideneacetone) palladium

Pd(dppf)Cl ₂ 1,1-bis(diphenylphosphino)ferrocenepalladium dichloride

Pd(OAc) ₂ palladium acetate

Pd(OH) ₂ palladium hydroxide

Pd(PPh ₃ ) _4tetrakis (triphenylphosphine)palladium

Pd(PPh ₃ ) ₂ Cl ₂ bis(triphenylphosphine)palladium dichloride

PE petroleum ether (60-90℃)

POC1 ₃ phosphorus oxychloride

PyBop 1H-Benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate

RT, rt, r.t. room temperature

Rt retention time

TBAB Tetrabutylammonium bromide

TBAHSO ₄ Tetrabutylammonium bisulfate

TBTU O-(1H-benzotriazol-1-yl)-N,N,N′,N′-tetramethylurea tetrafluoroborate

TFA trifluoroacetic acid

TEAC bis(tetraethylammonium) carbonate

THF Tetrahydrofuran

μL Microliter

The following synthetic schemes describe the steps involved in preparing the compounds disclosed in this invention. Unless otherwise stated, each of X, Z, R ⁵ , and R ⁶ has the definition as described in the present invention, V is an optionally substituted heterocycle or spirobicycle, and R ⁷ is H or methyl.

Synthetic method 1

Some compounds defined by formula (I) can be prepared by synthetic method 1:

(R)-aryl alcohol ( 1 ) reacts with substituted fluoropyridine ( 2 ) with NaH as the base in an aprotic solvent such as THF to obtain the coupling product ( 3 ). The nitro group on the pyridine ring in compound ( 3 ) is converted into amino compound ( 4 ) using a reducing agent such as Fe powder under acidic reducing conditions. Then, under the action of NBS, compound ( 5 ) was obtained by regioselective bromination on the pyridine ring. Finally, compound ( 5 ) is coupled with alkyne ( 6 ), such as 2-methylbut-3-yn-2-ol, under the action of a suitable Pd catalyst, to generate kinase inhibitor ( 7 ), which is defined by formula (I) compound of.

Synthetic method 2

Other compounds in the present invention can be prepared by synthetic method 2:

According to the method in Synthesis Method 1 or other methods, the compound represented by formula ( 5 ) can be prepared. Compound ( 5 ) and Boc anhydride can generate compound ( 8 ) under the action of a base such as triethylamine. The Sonogashira coupling between compound ( 8 ) and acetylenic alcohol ( 9 ) in the presence of a suitable Pd catalyst such as bis(triphenylphosphine)palladium dichloride gives compound ( 10 ). Afterwards, compound ( 10 ) was transformed into compound ( 11 ) under basic conditions. The obtained compound ( 11 ) undergoes nucleophilic substitution with nitrogen-containing heterocycle or azaspirobicyclic compound ( 12 ) to generate compound ( 13 ). Finally, under acidic conditions, such as trifluoroacetic acid in dichloromethane solution, hydrogen chloride in ethyl acetate solution, the amino protecting group (Boc) is removed to obtain the target product ( 14 ), which is the compound defined by formula (I).

The present invention adopts the following method to carry out biological test to the compound shown in formula (I):

1. Bioanalytical methods

The LC/MS/MS system used for analysis includes Agilent1200 series vacuum degassing furnace, binary syringe pump, orifice autosampler, column oven, and AgilentG6430 triple quadrupole mass spectrometer with electrospray ionization (ESI) source. Quantitative analysis was carried out in MRM mode, and the parameters of MRM conversion are shown in Table A:

Table A

multiple reaction detection scan 490.2 → 383.1 Fragmentation voltage 230V capillary voltage 55V Dryer temperature 350 ^°C atomizer 40psi Dryer flow rate 10L/min

The analysis uses Agilent XDB-C18, 2.1x30mm, 3.5 μM column, injecting 5 μL of sample. Analysis conditions: mobile phase is 0.1% formic acid aqueous solution (A) and 0.1% formic acid methanol solution (B). The flow rate was 0.4 mL/min. The mobile phase gradient is shown in Table B:

Form B

time Gradient of mobile phase B 0.5min 5% 1.0min 95% 2.2min 95%

2.3min 5% 5.0min stop

In addition, Agilent6330 series LC/MS/MS spectrometer was used for analysis, equipped with G1312A binary syringe pump, G1367A automatic sampler and G1314C UV detector; LC/MS/MS spectrometer used ESI radiation source. Optimum analysis is achieved using standards with appropriate cation model processing and MRM conversion for each analyte. A Capcell MP-C18 column, 100x4.6 mm I.D., 5 μM (Phenomenex, Torrance, California, USA) was used during the analysis. The mobile phase is 5mM ammonium acetate, 0.1% methanol in water (A): 5mM ammonium acetate, 0.1% methanol in acetonitrile (B) (70:30, v/v); the flow rate is 0.6mL/min; the column temperature is kept at room temperature; Inject 20 μL of sample.

2. Stability of compounds in human and rat liver microsomes

(1) Incubate human or rat liver microsomes in polypropylene test tubes and guide their replication. A typical incubation mixture consists of human or rat liver microsomes (0.5 mg protein/mL), target compound (5 μM), and NADPH (1.0 mM) potassium phosphate buffer (PBS, 100 mM, pH 7.4) in a total volume of 200 μL ), the compounds were dissolved in DMSO and diluted with PBS so that the final concentration of the DMSO solution was 0.05%. And incubate at 37° C. in a water bath connected to air. After pre-incubation for 3 minutes, add protein to the mixture and start the reaction. At different time points (0, 5, 10, 15, 30 and 60 min), the same volume of ice-cold acetonitrile was added to terminate the reaction. Samples were stored at -80°C until LC/MS/MS analysis.

Compound concentrations in human or rat liver microsome incubation mixtures were determined by LC/MS/MS. The linear range of the concentration range was determined for each compound tested.

Parallel incubation experiments using denatured microsomes as negative controls were incubated at 37°C and the reactions were terminated at different time points (0, 15 and 60 min).

Dextromethorphan (70 μM) was used as a positive control, incubated at 37°C, and the reactions were terminated at different time points (0, 5, 10, 15, 30 and 60 minutes). Positive and negative control samples are included with each assay to ensure the integrity of the microsomal incubation system.

(2) The stability data of the compounds of the present invention in human or rat liver microsomes can also be obtained from the following tests. Human or rat liver microsomes are incubated in polypropylene tubes and primed for replication. A typical incubation mixture includes human or rat liver microsomes (final concentration: 0.5mg protein/mL), compound (final concentration: 1.5μM) and K-buffer solution (containing 1.0mM EDTA, 100mM, pH7 .4). Compounds were dissolved in DMSO and diluted with K-buffer solution so that the final concentration of DMSO was 0.2%. After pre-incubation for 10 minutes, 15 μL NADPH (final concentration: 2 mM) was added for enzymatic reaction, and the whole experiment was carried out in an incubation tube at 37°C. At various time points (0, 15, 30 and 60 minutes), the reaction was terminated by adding 135 μL of acetonitrile (with IS). Centrifuge at 4000rpm for 10 minutes to remove protein, collect supernatant and analyze by LC-MS/MS.

In the above experiments, ketanserin (1 μM) was selected as a positive control, incubated at 37°C, and the reaction was terminated at different time points (0, 15, 30 and 60 minutes). A positive control sample is included with each assay to ensure the integrity of the microsomal incubation system.

The present invention adopts following method to carry out data analysis, to obtain stability analysis result:

For each reaction, the in vivo liver intrinsic clearance CL _int (ref.:Naritomi Y, Terashita S, Kimura S, Suzuki A, Kagayama A, Sugiyama Y. Prediction of human hepatic clearance from in vivo animal experiments and in vitro metabolic studies with liver microsomes from animals and humans. Drug Metabolism and Disposition 2001, 29: 1316-1324.)

3. Pharmacokinetic evaluation of the compounds of the present invention in animals

The present invention uses the following methods to evaluate the pharmacokinetic studies of the compounds of the present invention in mice, rats, dogs or monkeys:

The compound of the present invention is administered in the form of aqueous solution or 2% HPMC + 1% Tween-80 aqueous solution, 5% DMSO + 5% saline solution, 4% MC or capsule form. For intravenous administration, animals were dosed at 1 or 2 mg/kg. Oral doses (p.o.) were 5 or 10 mg/kg for rats and mice, and 10 mg/kg for dogs and monkeys. Blood (0.3 mL) was drawn at time points 0.25, 0.5, 1.0, 2.0, 3.0, 4.0, 6.0, 8.0, 12 and 24 hours and centrifuged at 3,000 or 4,000 rpm for 10 minutes. Plasma solutions were collected and stored at -20°C or -70°C until LC/MS/MS analysis as described above.

4. Kinase test

Kinase assays are performed by detecting myelin basic protein (MBP) incorporated into γ- ^33P -ATP. Prepare 20 μg/ml of MBP (Sigma#M-1891) tris-buffered saline solution (TBS; 50mM Tris pH8.0, 138mM NaCl, 2.7mM KCl), and coat a highly binding white 384-well plate (Greiner ), 60 μL per well. 4°C, incubate for 24h. The plate was then washed 3 times with 100 μL TBS. Kinase reactions were performed in a total volume of 34 μL in kinase buffer (5 mM Hepes pH 7.6, 15 mM NaCl, 0.01% bovine serum albumin (Sigma #I-5506), 10 mM MgCl ₂ , 1 mM DTT, 0.02% TritonX-100) . Compounds were dissolved in DMSO and added to the wells at a final concentration of 1% DMSO. Each data was determined twice, and the determination of each compound was carried out at least twice. For example, the final concentration of the enzyme is 10 nM or 20 nM. The reaction was started by adding unlabeled ATP (10 μM) and γ- ³³ P-labeled ATP (2×10 ⁶ cpm per well, 3000 Ci/mmol). The reaction was carried out with shaking at room temperature for 1 hour. The 384-well plate was washed with 7x PBS, and then 50 μL of scintillation fluid was added to each well. Results were checked with a Wallac Trilux counter. For those skilled in the art, this is only one of many detection methods, and other methods are also acceptable.

IC ₅₀ of inhibition and/or inhibition constant K _i can be obtained by the above test method. _IC50 is defined as the concentration of the compound that inhibits 50% of the enzyme activity under the test conditions. Use a 1/2 log dilution to generate a curve containing 10 concentration points to estimate the IC ₅₀ value (for example, a typical curve is generated by the following compound concentrations: 10 μM, 3 μM, 1 μM, 0.3 μM, 0.1 μM, 0.03 μM, 0.01μM, 0.003μM, 0.001μM, 0μM).

The kinase test in the present invention is done by Millipore, UK (MilliporeUK Ltd, Dundee Technology Park, Dundee DD21SW, UK).

4.1 ALK(h) kinase assay

Human ALK was incubated in the presence of 8mM MOPS with a pH value of 7.0, 0.2mM EDTA, 250μM KKKSPGEYVNIEFG, 10mM magnesium acetate and [γ-33P-ATP] (specific activity about 500cpm/pmol, concentration determined according to requirements). The reaction was initiated after the addition of the MgATP mixture. After incubation at room temperature for 40 minutes, a 3% phosphoric acid solution was added thereto to stop the reaction. 10 μL of the reaction solution was spotted onto a P30 filter, washed 3 times in 5 min with 75 mM phosphoric acid, and stored in methanol solution immediately prior to drying and scintillation counting.

4.2 c-Met(h) kinase assay

Human c-Met was incubated in the presence of 8mM MOPS with a pH value of 7.0, 0.2mM EDTA, 250μM KKKSPGEYVNIEFG, 10mM magnesium acetate and [γ- ³³ P-ATP] (specific activity about 500cpm/pmol, concentration determined according to requirements) . The reaction was initiated after the addition of the MgATP mixture. After incubation at room temperature for 40 minutes, a 3% phosphoric acid solution was added thereto to stop the reaction. 10 μL of the reaction solution was spotted onto a P30 filter, washed 3 times in 5 min with 75 mM phosphoric acid, and stored in methanol solution immediately prior to drying and scintillation counting.

5. Cell phosphorylation test

Typically, cells are preincubated with the compound to be tested to allow for sufficient target binding. The level of autophosphorylation was detected by sandwich enzyme-linked immunoassay (Sandwich-ELISA). A curve containing 8 concentration points was made using a dilution factor of 1/1log to estimate the IC ₅₀ value (measured twice for each concentration). The cell phosphorylation test in the present invention is done by ProQinase GmbH (ProQinase GmbH, Breisacher Straβe117D-79106, Freiburg, Germany).

5.1 c-Met phosphorylation assay

It is well known that the human gastric adenocarcinoma cell line MKN45 overexpresses c-Met. c-Met overexpression leads to constitutive, ligand-independent kinase autophosphorylation. With the addition of SU11274, the level of phosphorylated Met was significantly reduced, thus confirming the inhibitory ability of the compounds of the present invention. Phosphorylated Met signal can be quantified by sandwich enzyme-linked immunoassay (Sandwich-ELISA) technique. The test set up a known Met inhibitor group to verify the reliability of the test method.

6. Xenograft tumor model

The drug effect of the compound of the present invention is evaluated by the standard mouse model of implanted tumor, the method is as follows:

Human tumor cells (for example, U87MG neuroblastoma cells, MKN45 gastric adenocarcinoma cells, MDA-MB-231 breast cancer cells, or Caki-1 kidney cancer cells, cells from ATCC Company) were cultured and collected, and placed on the ventral side Inoculate subcutaneously in 6-7 week old female nude mice (BALB/cA nu/nu, Shanghai SLAC Animal Laboratory) (n=10 for the vehicle group, n=8 for each dose group). When the tumor volume reached 100-250 mm ³ , the animals were randomly divided into solvent control group (2% HPMC+1% Tween-80 aqueous solution) and compound group. Subsequent intragastric administration of compounds (3-50 mpk/dose, dissolved in 2% HPMC + 1% Tween-80 in water) to animals, starting anywhere from 0 to 15 days after tumor cell inoculation, And usually once a day in the trial.

6.1 Tumor Growth Inhibition (TGI) Analysis

Tumor evolutionary growth was assessed by tumor volume versus time. The long axis (L) and short axis (W) of subcutaneous tumors were measured twice a week by calipers, and the tumor volume (TV) was calculated by the formula (L×W ² )/2). TGI is calculated from the difference between the median tumor volume of the mice in the solvent group and the median tumor volume of the mice in the drug group, expressed as a percentage of the median tumor volume of the solvent control group, and calculated by the following formula:

Raw statistical analysis was done by repeated measures of variance (RMANOVA). Next, multiple comparisons were performed by the Scheffe psot hoc test method. Solvent alone (2% HPMC + 1% Tween-80, etc.) served as a negative control.

The results show that the compound provided by the invention exhibits good half-life and good pharmacokinetic properties, has good inhibitory effect on ALK and c-Met, and also has good inhibitory effect on tumor growth.

The compounds, pharmaceutical compositions and applications provided by the present invention will be further described below in conjunction with the examples.

Example 1: 4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)butan-3- Alkyn-2-ol

Step 1) (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-2-nitropyridine

Dissolve (R)-1-(2,6-dichloro-3-fluorophenyl)ethanol (10g, 47.81mmol) in tetrahydrofuran (10mL), cool to 0°C, and add to it in batches within 30 minutes Sodium hydride (1.4 g, 57.42 mmol) was added. After stirring at room temperature for 2 hours, it was cooled to 0°C again, and a solution of 3-fluoro-2-nitropyridine (8.2 g, 57.43 mmol) in tetrahydrofuran (80 mL) was added dropwise to the reaction system within 20 minutes. Return to room temperature and continue stirring for 3 hours. After the reaction was completed, the reaction was quenched with ice water (10 mL), and concentrated under reduced pressure. The residue was diluted with water (150 mL), and extracted with ethyl acetate (150 mL x 3). The combined organic phases were washed successively with saturated NaHCO ₃ (400 mL) and saturated brine (400 mL), and dried over anhydrous Na ₂ SO ₄ . It was concentrated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (PE/EtOAc (v/v)=6:1 to 4:1) to obtain the title compound as a white solid (13.4 g, 84.8%).

LC-MS (ESI, pos.ion) m/z: 331 [M+H] ⁺ .

Step 2) (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-2-amine

(R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-2-nitropyridine (13.4 g, 39.3 mmol) was suspended in ethanol (250 mL), and Iron powder (11 g, 197 mmol) was added thereto. After the mixture was stirred at 90°C for 20 minutes, 1N HCl (4 mL) was added to the reaction mixture, stirring was continued for 15 minutes, and 1N HCl (4 mL) was added again. The reaction solution was stirred at 90°C for 2 hours. After the reaction was completed, it was cooled to room temperature, the mixture was filtered with celite, and the filter cake was washed with ethanol (80mL x 3). The filtrate was concentrated under reduced pressure to afford the title compound as a light brown solid (12 g, 100%).

LC-MS (ESI, pos.ion) m/z: 301[M+H] ⁺ ;

¹ H NMR(400MHz,d ₆ -DMSO)δ(ppm):1.75(d,J=6.6Hz,3H),5.67(brs,2H),5.97-5.92(q,J=6.6Hz,1H),6.38 -6.35(dd,J=7.7Hz,5.0Hz,1H),6.61(d,J=7.1Hz,1H),7.47-7.42(m,2H),7.56-7.52(dd,J=7.7Hz,5.0Hz ,1H).

Step 3) (R)-5-bromo-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-2-amine

Dissolve (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-2-amine (12g, 39.3mmol) in acetonitrile (250mL) and cool to 0 °C, NBS (9.4 g, 52.3 mmol) was added thereto in portions within 20 minutes. After the reaction solution was stirred at 0°C for 1 hour, it was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (PE/EtOAc (v/v)=4/1 to 3/1) to obtain the title compound as a light brown solid ( 10g, 68%).

LC-MS (ESI, pos.ion) m/z: 379[M+H] ⁺ ;

¹ H NMR(400MHz,d ₆ -DMSO)δ(ppm):1.82(d,J=6.6Hz,3H),4.82(brs,2H),6.01-5.96(q,J=6.6Hz,1H),6.83 (d,J=1.8Hz,1H),7.10(t,J=8.0Hz,1H),7.33-7.30(dd,J=8.9Hz,4.8Hz,2H),7.66(dd,J=5.0Hz,1.8 Hz, 1H).

Step 4) 4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)but-3-yne -2-ol

(R)-5-bromo-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-2-amine (0.19g, 0.5mmol), bis(triphenyl Phosphorus) palladium dichloride (35mg, 0.05mmol) and cuprous iodide (9.5mg, 0.05mmol) were suspended in triethylamine (2.5mL), and 3-butyn-2-ol (53mg, 0.75 mmol). The reaction solution was microwaved at 90° C. for 80 minutes, cooled to room temperature, and concentrated under reduced pressure. The residue was diluted with dichloromethane (10 mL) and filtered, and the filtrate was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM/MeOH (v/v)=10/1) to obtain the title compound as a yellow solid (0.1 g, 54%).

LC-MS (ESI, pos.ion) m/z: 369[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ) δ (ppm): 1.50-1.52 (m, 3H), 1.81 (d, J=6.7Hz, 3H), 4.68-4.73 (m, 1H), 5.02 (s, 2H) ,5.98-6.03(q,J=6.6Hz,1H),6.74(d,J=1.4Hz,1H),7.05-7.09(m,1H),7.26-7.32(m,1H),7.77(d,J =1.0Hz,1H).

Example 2: (R)-4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-2- Methylbut-3-yn-2-ol

(R)-5-bromo-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-2-amine (0.19g, 0.5mmol), bis(triphenyl Phosphine)palladium dichloride (35mg, 0.05mmol) and cuprous iodide (9.5mg, 0.05mmol) were suspended in triethylamine (2.5mL), and 2-methylbut-3-yn-2 was added thereto - Alcohol (63 mg, 0.75 mmol). The reaction solution was microwaved at 90° C. for 1 hour, cooled to room temperature, and concentrated under reduced pressure. The residue was diluted with dichloromethane (10 mL) and filtered, and the filtrate was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM/MeOH/Et ₃ N (v/v/v)=1000/50/1) to obtain the title compound as a white solid (115 mg, 81%).

LC-MS (ESI, pos.ion) m/z: 384[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ) δ (ppm): 1.63 (s, 6H), 1.80 (d, J=6.4Hz, 3H), 2.46 (1H), 4.97 (s, 2H), 6.01 (q, J =6.4Hz,1H),6.74(d,J=1.56Hz,1H),7.07(m,1H),7.29(m,1H),7.74(s,1H);

¹³ C NMR (100MHz, CDCl ₃ ) δ (ppm): 18.8, 29.7, 31.5, 65.5, 76.7, 93.8, 108.7, 116.8, 119.3, 128.9, 130.0, 136.8, 138.8, 143.0, 150.2.

Example 3: (R)-5-((1H-pyrazol-4-yl)ethynyl)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy) Pyridin-2-amine

Step 1) 4-((Trimethylsilyl)ethynyl)-1H-pyrazole

4-iodo-1H-pyrazole (2.91g, 15mmol), bis(triphenylphosphine) palladium dichloride (1g, 1.5mmol), cuprous iodide (286mg, 1.5mmol) and triethylamine (6mL ) was dissolved in ethanol (20 mL), and trimethylsilylacetylene (3.2 mL, 22.5 mmol) was added thereto. The reaction solution was stirred at 70°C for 4 hours, cooled to room temperature, and concentrated under reduced pressure. The residue was diluted with ethyl acetate (20 mL), filtered, the filtrate was dried over anhydrous sodium sulfate, and concentrated under reduced pressure, the residue was purified by silica gel column chromatography (PE/EtOAc (v/v)=5/1), The title compound was obtained as a white solid (1.3 g, 53%).

LC-MS (ESI, pos.ion) m/z: 165[M+H] ⁺ ;

¹ H NMR (400 MHz, CDCl ₃ ) δ (ppm): 0.24 (s, 9H), 7.76 (s, 2H), 11.43 (br, 1H).

Step 2) 4-ethynyl-1H-pyrazole

4-((Trimethylsilyl)ethynyl)-1H-pyrazole (1 g, 6.1 mmol) was dissolved in tetrahydrofuran (12 mL) and water (2 mL), and potassium carbonate (1 g, 12.2 mmol) was added thereto . After the reaction solution was stirred at room temperature for 2 hours, it was concentrated under reduced pressure to obtain the title compound, which was directly used in the next step without purification.

LC-MS (ESI, pos.ion) m/z: 93[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ) δ (ppm): 3.04(s, 1H), 7.75(s, 2H), 10.85(br, 1H).

Step 3) (R)-5-((1H-pyrazol-4-yl)ethynyl)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridine Pyridine-2-amine

(R)-5-bromo-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-2-amine (695mg, 1.83mmol), bis(triphenylphosphine ) palladium dichloride (128mg, 0.18mmol) and cuprous iodide (35mg, 0.18mmol) were dissolved in ethanol (12mL) and dioxane (12mL), and N,N-diisopropyl Ethylamine (4 mL) and 4-ethynyl-1H-pyrazole (168 mg, 1.83 mmol). The reaction solution was stirred at 80°C for 16 hours, cooled to room temperature, and concentrated under reduced pressure. The residue was diluted with dichloromethane (25 mL), filtered, and the filtrate was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM/MeOH/Et ₃ N (v/v/v)=1000/50/1) to give the title compound as a white solid (0.25, 35%).

LC-MS (ESI, pos.ion) m/z: 393[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ )δ(ppm): 1.84(d, J=6.8Hz, 1H), 6.09(q, J=6.8Hz, 1H), 6.73(d, J=1.6Hz, 1H), 7.25(m,1H),7.46(m,1H),7.60(d,J=1.68Hz,1H),7.62(s,1H),7.83(s,1H);

¹³ C NMR (100 MHz, d ₆ -DMSO) δ (ppm): 18.7, 72.2, 80.8, 87.3, 101.2, 106.8, 117.4, 118.1, 120.8, 128.5, 130.5, 136.5, 137.8, 142.5, 150.8, 155.5, 158.0.

Example 4: 5-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridine-3- base)-1,1,1-trifluoro-2-methylpent-4-yn-2-ol

Step 1) 1,1,1-Trifluoro-2-methylpent-4-yn-2-ol

Aluminum foil (0.34 g, 12.6 mmol) was suspended in tetrahydrofuran (6 mL), and a small amount of mercury chloride (5 mg, 0.02 mmol) was added thereto. After the mixture was stirred vigorously at room temperature for 1 hour, propargyl bromide (1.5 g, 12.6 mmol) was added dropwise thereto over 30 minutes. The reaction solution was stirred at 40° C. for 2 hours, cooled to room temperature, and stirred for 3 hours, and set aside;

1,1,1-Trifluoroacetone (0.24g, 2.09mmol) was dissolved in ether (20mL), cooled to -78°C, and the suspension above was added thereto. The reaction was returned to room temperature and stirred at room temperature for 16 hours before being quenched with water (30 mL). The organic phase was separated, the aqueous phase was extracted with ether (40mL x3), the combined organic phase was washed with saturated brine (100mL x2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure (concentration temperature<5°C) to obtain the title compound as Pale yellow solid (0.46g, >100%). The crude product was directly used in the next reaction without purification.

GC-MS(EI): 152.0, 137.0, 126.8, 113.0.

Step 2) 5-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1,1,1- Trifluoro-2-methylpent-4-yn-2-ol

(R)-5-bromo-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-2-amine (0.3g, 0.79mmol), bis(triphenyl Phosphine)palladium dichloride (27mg, 0.04mmol) and cuprous iodide (7mg, 0.04mmol) were suspended in N,N-dimethylformamide (5mL), and 1,1,1-tris Fluoro-2-methylpent-4-yn-2-ol (0.32 g, 2.1 mmol) in N,N-dimethylformamide (5 mL) and triethylamine (2.2 mL, 15.8 mmol). The reaction solution was heated at 75°C for 16 hours, cooled to room temperature, and diluted with dichloromethane (150 mL). The resulting mixture was washed successively with water (100 mL x2), saturated brine (100 mL x2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE/EtOAc(v/v)=2/1) and preparative silica gel plate (PE/EtOAc(v/v)=2/1) successively to obtain the title compound as a pale yellow solid ( 0.16g, 45%).

LC-MS (ESI, pos.ion) m/z: 451.1[M+H] ⁺ ;

¹ H NMR(400MHz,CDCl ₃ )δ(ppm):1.49(s,3H),1.81(d,J=6.68Hz,3H),2.72(d,J=17.00Hz,1H),2.89(d,J =17.02Hz,1H),5.01(s,2H),5.97-6.03(q,J=6.68Hz,1H),6.72(m,1H),7.10(t,J=8.48Hz,1H),7.27-7.52 (dd, J=8.92Hz, 4.84Hz, 1H), 7.72(m, 1H).

Example 5: 4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridine-3- base)-1,1,1-trifluoro-2-methylbut-3-yn-2-ol

Step 1) 1,1,1-Trifluoro-2-methyl-4-(trimethylsilyl)but-3-yn-2-ol

Trimethylsilylacetylene (1 g, 10.2 mmol) was dissolved in ether (10 mL), cooled to -78°C, and n-butyllithium (4.11 mL, 10.3 mmol) was added thereto by syringe within 30 minutes. After the mixture was stirred at -78°C for 30 minutes, a solution of 1,1,1-trifluoroacetone (1.37 mL, 10.2 mmol) in diethyl ether (10 mL) was further added thereto within 30 minutes. The reaction solution was stirred at room temperature for 50 minutes, then quenched with saturated aqueous ammonium chloride solution (20 mL), the organic phase was separated, and the aqueous phase was extracted with diethyl ether (30 mL x 4). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure (<10°C) to give the title compound as a pale yellow oil (2.1 g, 98%).

GC-MS (EI): 209.0, 137.0, 113.0.

Step 2) 1,1,1-Trifluoro-2-methylbut-3-yn-2-ol

1,1,1-Trifluoro-2-methyl-4-(trimethylsilyl)but-3-yn-2-ol (0.8 g, 3.8 mmol) was dissolved in methanol (10 mL), and To this was added potassium hydroxide (1.28 g, 22.8 mmol). After the reaction solution was stirred at room temperature for 16 hours, it was quenched with water (30 mL), and extracted with ether (40 mL×3). The combined organic phases were washed with saturated brine (100mL x2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure (<10°C), and the residue was dried under vacuum at 5°C to obtain the title compound as a colorless oil (0.31g, 59%).

GC-MS (EI): 137.0, 113.0.

Step 3) 4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1,1,1- Trifluoro-2-methylbut-3-yn-2-ol

Dissolve (R)-5-bromo-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-2-amine (0.2 g, 0.53 mmol) in dichloromethane , under the protection of N ₂ , bis(triphenylphosphine)palladium dichloride (18mg, 0.03mmol), cuprous iodide (5mg, 0.03mmol), 1,1,1-trifluoro-2 - Methylbut-3-yn-2-ol (0.31 g, 2.37 mmol) and triethylamine (1.47 mL, 10.5 mmol). The reaction solution was stirred at 75°C for 2.5 hours, cooled to room temperature, and diluted with dichloromethane (150 mL). The resulting mixture was washed with saturated brine (200 mL x 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by preparative silica gel plate to afford the title compound as a yellow solid (30 mg, 13%).

LC-MS (ESI, pos.ion) m/z: 437.1[M+H] ⁺ ;

¹ H NMR(400MHz, CDCl ₃ )δ(ppm):1.22(s,3H),1.75(d,J=6.56Hz,3H),5.96-6.02(q,J=6.68Hz,1H),6.29(s ,2H),6.59(s,1H),7.50(t,J=8.60Hz,1H),7.54-7.59(dd,J=8.92Hz,5.00Hz,1H),7.62(s,1H).

Example 6: (R)-4-((6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)acetylene Base) piperidin-4-ol

Step 1) 1-(tert-butoxycarbonyl)-4-((trimethylsilyl)ethynyl)piperidin-4-ol

Under nitrogen protection, trimethylsilylacetylene (1.7mL, 12mmol) was dissolved in anhydrous tetrahydrofuran (20mL), cooled to -40°C, n-butyllithium solution (4.8mL, 12mmol) was slowly added to the solution, After the mixture was stirred at -40°C for 1 hour, it was cooled to -78°C, and 1-(tert-butoxycarbonyl)piperidin-4-one (2 g, 10 mmol) in anhydrous tetrahydrofuran (10 mL ) solution. The reaction solution was stirred at -78°C for 1 hour, then returned to room temperature, and stirred at room temperature for 72 hours. After completion of the reaction, add saturated ammonium chloride aqueous solution (15mL) and water (50mL) to quench, and extract with ethyl acetate (50mL x3), the combined organic phase is washed with water (50mL) and saturated brine (50mL) successively, and anhydrous Dry over sodium sulfate, and concentrate under reduced pressure. The residue was purified by column chromatography (PE/EtOAc (v/v)=5/1) to give the title compound as a white solid (1.93 g, 65%).

¹ H NMR (400MHz, CDCl ₃ ) δ (ppm): 0.17 (s, 9H), 1.46 (s, 9H), 1.68 (m, 2H), 1.87 (m, 2H), 2.19 (s, 1H), 3.22 (m,2H),3.79(br,2H);

¹³ C NMR (100 MHz, CDCl ₃ ) δ (ppm): 28.5, 39.0, 67.2, 79.6, 89.9, 107.7, 154.7.

Step 2) 1-(tert-butoxycarbonyl)-4-ethynylpiperidin-4-ol

1-(tert-butoxycarbonyl)-4-((trimethylsilyl)ethynyl)piperidin-4-ol (1.4 g, 6.22 mmol) was dissolved in methanol (12 mL) and water (2 mL), and Potassium carbonate (3.4 g, 24.88 mmol) was added thereto. After stirring the reaction solution at room temperature for 2 hours, it was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE/EtOAc (v/v)=5/1) to give the title compound as a white solid (0.95 g, 90%).

¹ H NMR (400MHz, CDCl ₃ ) δ (ppm): 1.46 (s, 9H), 1.68 (m, 2H), 1.87 (m, 2H), 2.53 (s, 1H), 3.27 (m, 2H), 3.50 (s,1H),3.75(br,2H);

¹³ C NMR (100 MHz, CDCl ₃ ) δ (ppm): 28.6, 38.9, 40.8, 66.5, 73.1, 79.9, 86.5, 154.8.

Step 3) (R)-4-((6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)acetylene Base)-1-(tert-butoxycarbonyl)piperidin-4-ol

(R)-5-bromo-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-2-amine (695mg, 1.83mmol), bis(triphenylphosphine ) palladium dichloride (128mg, 0.18mmol), cuprous iodide (35mg, 0.18mmol) and potassium carbonate (505mg, 3.66mmol) were suspended in dioxane (12mL), and N,N- Diisopropylethylamine (4 mL) and 1-(tert-butoxycarbonyl)-4-ethynylpiperidin-4-ol (412 mg, 1.83 mmol). The reaction solution was stirred at 90°C for 16 hours, cooled to room temperature, and concentrated under reduced pressure. The residue was diluted with dichloromethane (20 mL), and filtered. The filtrate was dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE/EtOAc (v/v)=1/1) to obtain the title compound as a pale yellow solid (0.15 g, 16%).

LC-MS (ESI, pos.ion) m/z: 524[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ) δ (ppm): 1.48 (s, 9H), 1.76 (m, 2H), 1.81 (d, J=6.8Hz, 1H), 1.92 (m, 2H), 3.25 (m ,2H),3.82(br,2H),5.04(br,2H),6.73(s,1H),6.99(q,J=6.8Hz,1H),7.30(m,1H),7.74(m,1H) ,7.79(br,1H);

¹³ C NMR (100MHz, CDCl ₃ ) δ (ppm): 14.1, 28.6, 39.1, 67.1, 72.6, 82.9, 91.5, 116.8, 119.2, 122.0, 128.9, 130.0, 136.6, 154.8.

Step 4) (R)-4-((6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)ethynyl) piperidin-4-ol

(R)-4-((6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)ethynyl)-1-(tert Butoxycarbonyl)piperidin-4-ol (150mgm, 0.29mmol) was dissolved in dichloromethane (4mL), cooled to 0°C, and hydrogen chloride in ethyl acetate (3M, 10mL) was added to the solution. After the reaction solution was stirred at room temperature for 4 hours, the product precipitated and was filtered. Water (10 mL) was added to the obtained solid, and the pH was adjusted to 10 with 4M aqueous sodium hydroxide solution. The resulting mixture was extracted with ethyl acetate (10 mL x 3), the combined organic phases were dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM/MeOH/Et ₃ N (v/v/v)=1000/50/1) to give the title compound as a white solid (70 mg, 56%).

LC-MS (ESI, pos.ion) m/z: 424[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ) δ (ppm): 1.76 (m, 2H), 1.81 (d, J=6.8Hz, 1H), 1.92 (m, 2H), 2.86 (m, 2H), 3.12 (br ,2H),4.96(br,2H),5.99(q,J=6.8Hz,1H),6.73(s,1H),7.30(m,1H),7.74(m,1H),7.76(s,1H) ;

¹³ C NMR (100MHz, CDCl ₃ ) δ (ppm): 11.5, 40.7, 43.7, 67.6, 72.6, 82.7, 91.9, 116.8, 119.3, 120.2, 128.9, 136.8, 138.8, 142.9, 150.2.

Example 7: 3-((6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)acetylene Base) Tetrahydrofuran-3-ol

Step 1) 3-((Trimethylsilyl)ethynyl)tetrahydrofuran-3-ol

Trimethylsilylacetylene (1 g, 10.2 mmol) was dissolved in THF (20 mL), cooled to -78°C, and n-butyllithium (4.11 mL, 10.3 mmol) was added thereto. After the mixture was stirred at -78°C for 30 minutes, a solution of dihydrofuran-3(2H)-one (0.87 g, 10.2 mmol) in tetrahydrofuran (20 mL) was continuously added to the system. The reaction solution was stirred at -78°C for 30 minutes, returned to room temperature, and stirred for 50 minutes. After the reaction was completed, it was quenched by adding saturated aqueous ammonium chloride (20 mL), and extracted with ethyl acetate (50 mL x 4). The combined organic phases were washed with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the title compound as a white solid (1.67 g, 89%). The product was directly used in the next reaction without purification.

¹ H NMR(400MHz,d ₆ -DMSO)δ(ppm):0.14(s,9H),2.05-2.08(m,2H),3.62-3.70(m,2H),3.79-3.83(m,2H), 5.72(s,1H).

Step 2) 3-Ethynyltetrahydrofuran-3-ol

3-((Trimethylsilyl)ethynyl)tetrahydrofuran-3-ol (1.67 g, 9.06 mmol) was dissolved in methanol (50 mL), and potassium hydroxide (1.01 g, 18.12 mmol) was added thereto. After the reaction solution was stirred at room temperature for 2 hours, it was washed with water (100mLx2) and saturated brine (100mLx2) successively. The mixture was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the title compound as a yellow oil (0.77 g, 75.9%).

LC-MS (ESI, pos.ion) m/z: 113.05[M+H] ⁺ ;

¹ H NMR(400MHz,d ₆ -DMSO)δ(ppm):2.06-2.10(m,2H),3.41-3.42(m,1H),3.63-3.70(m,2H),3.80-3.83(m,2H ), 5.72(s,1H).

Step 3) 3-((6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)ethynyl) Tetrahydrofuran-3-ol

Under nitrogen protection, 3-ethynyltetrahydrofuran-3-ol (0.72g, 6.43mmol), (R)-5-bromo-3-(1-(2,6-dichloro-3-fluorophenyl)ethane Oxy)pyridin-2-amine (1.88g, 4.95mmol), bis(triphenylphosphine)palladium dichloride (0.17g, 0.25mmol), cuprous iodide (47mg, 0.25mmol) dissolved in N,N - Dimethylformamide (20 mL), and triethylamine (9.99 g) was added thereto. The reaction solution was stirred at 100°C for 12 hours, and then concentrated under reduced pressure. The residue was diluted with dichloromethane (100 mL), and washed successively with water (100 mL x2) and saturated brine (100 mL x2). The resulting mixture was dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE/EtOAc (v/v)=1/1), and recrystallized in a mixture of petroleum ether and ethyl acetate ((v/v)=10/1) to obtain The title compound was a pale yellow solid (0.3 g, 15%).

LC-MS (ESI, pos.ion) m/z: 412.00[M+H] ⁺ ;

¹ H NMR (400MHz,d ₆ -DMSO)δ(ppm):1.74-1.76(d,3H),2.10-2.13(m,2H),3.66-3.74(m,2H),3.82-3.86(m,2H ),5.74(s,1H),5.96-6.00(m,1H),6.16(s,2H),6.60-6.61(m,1H),7.44-7.49(m,1H),7.56-7.57(m,1H ),7.59-7.60(m,1H).

Example 8: 3-((6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)acetylene base) pyrrolidin-3-ol

Step 1) 1-(tert-butoxycarbonyl)-3-((trimethylsilyl)ethynyl)pyrrolidin-3-ol

Dissolve trimethylsilylacetylene (1g, 10.2mmol) in tetrahydrofuran (20mL), cool to -78°C, and add n-butyllithium (4.11mL, 10.3mmol, 2.5M n-hexane solution). After the mixture was stirred at -78°C for 30 minutes, a solution of N-Boc-3-pyrrolidone (1.89 g, 10.2 mmol) in tetrahydrofuran was further added thereto within 30 minutes. After the reaction solution was stirred at room temperature for 50 minutes, it was quenched by adding saturated aqueous ammonium chloride solution (8 mL), and extracted with ethyl acetate (50 mL x 4). The combined organic phases were washed with saturated brine (100ml x 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the title compound as a light yellow solid (2.6g, 90%).

¹ H NMR (400 MHz, CDCl ₃ ) δ (ppm): 0.17 (s, 9H), 1.45 (s, 9H), 2.10-2.24 (m, 2H), 3.44-3.67 (m, 4H).

Step 2) 1-(tert-butoxycarbonyl)-3-ethynylpyrrolidin-3-ol

1-(tert-butoxycarbonyl)-3-((trimethylsilyl)ethynyl)pyrrolidin-3-ol was dissolved in methanol (30 mL), and potassium hydroxide (0.59 g, 10.6 mmol ). After the reaction solution was stirred at room temperature for 2 hours, it was diluted with water (50 mL), and extracted with ethyl acetate (60 mL x 5). The combined organic phases were washed with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE/EtOAc (v/v)=3/1) to give the title compound as a white solid (0.86 g, 76%).

LC-MS (ESI, pos.ion) m/z: 156.2[M+H-56] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ )δ(ppm):1.45(s,9H),2.18(m,2H),2.58(m,1H),2.54(s,1H),3.47-3.72(m,4H) .

Step 3) 3-((6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)acetylene Base)-1-(tert-butoxycarbonyl)pyrrolidin-3-ol

Under _N2 protection, (R)-5-bromo-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-2-amine (0.3g, 0.79mmol) was dissolved In N,N-dimethylformamide (30mL), and bis(triphenylphosphine)palladium dichloride (28mg, 0.04mmol), CuI (7mg, 0.04mmol), 1-(tert Butoxycarbonyl)-3-ethynylpyrrolidin-3-ol (0.17 g, 0.79 mmol) and triethylamine (2.2 mL, 15.8 mmol). The reaction solution was stirred at 75°C for 16 hours, then cooled to room temperature, and diluted with dichloromethane (300 mL). The resulting mixture was washed with saturated brine (100 mL x 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE/EtOAc (v/v)=2/1 to 1/1) to obtain the title compound as a brown-red oil (0.25 g, 62%).

Step 4) 3-((6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)ethynyl) pyrrolidin-3-ol

3-((6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)ethynyl)-1-(tert Butoxycarbonyl)pyrrolidin-3-ol (0.25 g, 0.49 mmol) was dissolved in dichloromethane (20 mL), and saturated ethyl hydrogen chloride solution (5 mL) was added thereto. After stirring the reaction solution at room temperature for 16 hours, it was concentrated under reduced pressure. The residue was diluted with water (5 mL), adjusted to pH=12 with saturated aqueous sodium carbonate, and extracted with a mixture of DCM/MeOH (v/v=10:1) (40 mL x 5). The combined organic phases were washed with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by preparative silica gel plate (DCM/MeOH (v/v)=10/1) to afford the title compound as a yellow solid (0.11 g, 54%).

LC-MS (ESI, pos.ion) m/z: 410.0[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ +CD ₃ OD)δ(ppm): 1.83(d, J=6.68Hz, 3H), 2.25-2.48(m, 2H), 3.43-3.58(m, 4H), 5.88- 6.06(q,J=6.68Hz,1H),6.72(d,J=1.52Hz,1H),7.17(t,J=8.80Hz,1H),7.32-7.38(dd,J=8.92Hz,4.80Hz, 1H), 7.65 (d, J=1.56Hz, 1H).

Example 9: 3-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(3-morpholinobut-1-yn-1-yl) Pyridin-2-amine

Step 1) (R)-5-bromo-N,N-bis(tert-butoxycarbonyl)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridine Pyridine-2-amine

(R)-5-bromo-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-2-amine (2.52g, 6.63mmol), DMAP (809mg, 6.63 mmol) was dissolved in tetrahydrofuran (66 mL), and Boc anhydride (4.7 mL, 19.89 mmol) and triethylamine (2.8 mL, 19.89 mmol) were added thereto. After the reaction solution was stirred at 68°C for 11 hours, it was quenched by adding saturated aqueous sodium bicarbonate (50 mL), and extracted with ethyl acetate (50 mL x 3). The combined organic phases were washed successively with water (50 mL), saturated aqueous sodium bicarbonate solution (50 mL) and saturated brine (50 mL). The resulting solution was dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (PE/EtOAc (v/v)=4/1) to give the title compound as a yellow solid (4.4 g, 114%).

LC-MS (ESI, pos.ion) m/z: 603 [M+Na] ⁺ .

Step 2) 4-(6-(bis(tert-butoxycarbonyl)amino)-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy) Pyridin-3-yl)but-3-yn-2-ol

(R)-5-bromo-N, N-bis(tert-butoxycarbonyl)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-2-amine ( 0.25g, 0.43mmol), bis(triphenylphosphine) palladium dichloride (30mg, 0.04mmol) and cuprous iodide (8.1mg, 0.04mmol) were suspended in triethylamine (2.5mL), and 3-Butyn-2-ol (0.05 mL, 0.65 mmol) was added. The reaction solution was microwaved at 90° C. for 80 minutes, cooled to room temperature, and concentrated under reduced pressure. The residue was diluted with dichloromethane (10 mL), filtered, and the filtrate was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE/EtOAc (v/v)=2/1) to give the title compound as a yellow solid (201 mg, 82%).

¹ H NMR (400MHz, CDCl ₃ ) δ (ppm): 1.37-1.41 (m, 18H), 1.55 (d, J=6.6Hz, 3H), 1.78 (d, J=6.6Hz, 3H), 4.72-4.78 (m,1H),5.97-5.02(q,J=6.7Hz,1H),7.06-7.12(m,2H),7.27-7.32(m,1H),8.12-8.13(m,1H).

Step 3) 4-(6-(bis(tert-butoxycarbonyl)amino)-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy) Pyridin-3-yl)but-3-yn-2-methanesulfonate

Under nitrogen protection, 4-(6-(bis(tert-butoxycarbonyl)amino)-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridine- 3-yl)but-3-yn-2-ol (244mg, 0.43mmol) was dissolved in dichloromethane (2.4mL), cooled to -20°C, and triethylamine (0.14g, 1.38mmol) was added thereto and methanesulfonyl chloride (0.05 mL, 0.65 mmol). The reaction solution was stirred at -20°C for 3 hours, then returned to room temperature, and stirred for another 3 hours. The reaction was quenched by adding saturated aqueous sodium bicarbonate (10 mL), the resulting mixture was extracted with dichloromethane (10 mL x 3), the combined organic phases were dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the title compound as a yellow oil (243 mg , 99%), the product was directly used in the next step without purification.

LC-MS (ESI, pos.ion) m/z: 647 [M+H] ⁺ .

Step 4) N,N-bis(tert-butoxycarbonyl)-3-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(3-mol Linylbut-1-yn-1-yl)pyridin-2-amine

4-(6-(bis(tert-butoxycarbonyl)amino)-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl) But-3-yne-2-methanesulfonate (0.28 g, 0.43 mmol) was dissolved in acetonitrile (5 mL), and morpholine (0.13 g, 1.5 mmol) was added thereto. The reaction solution was stirred at 60°C for 16 hours, cooled to room temperature, and concentrated under reduced pressure. The residue was diluted with dichloromethane (50 mL), and the resulting mixture was washed with water (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE/EtOAc (v/v)=2/1) to obtain the title compound as a yellow solid (219 mg, 80%).

LC-MS (ESI, pos.ion) m/z: 638[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ) δ (ppm): 1.35-1.42 (m, 18H), 1.41-1.44 (m, 3H), 1.81 (d, J=6.7Hz, 3H), 2.54-2.56 (m, 2H),2.71-2.74(m,2H),3.63-3.69(m,1H),3.78-3.80(m,4H),5.98-6.03(q,J=6.7Hz,1H),7.06-7.12(m, 2H), 7.29-7.34 (m, 1H), 8.09 (s, 1H).

Step 5) 3-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(3-morpholinobut-1-yn-1-yl)pyridine Pyridine-2-amine

N,N-bis(tert-butoxycarbonyl)-3-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(3-morpholinobutyl -1-yn-1-yl)pyridin-2-amine (219 mg, 0.34 mmol) was dissolved in dichloromethane (5 mL), and ethyl hydrogen chloride acetate solution (3M, 1 mL) was added thereto. After the reaction solution was stirred at room temperature for 18 hours, it was quenched by adding saturated potassium carbonate aqueous solution (30 mL), and extracted with ethyl acetate (50 mL x 3). The combined organic phases were washed successively with water (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE/EtOAc (v/v)=1/1) to obtain the title compound as a yellow solid (134 mg, 90%).

LC-MS (ESI, pos.ion) m/z: 439[M+H] ⁺ ;

¹ H NMR(400MHz, CDCl ₃ )δ(ppm):1.37-1.39(m,3H),1.82(d,J=6.7Hz,3H),2.51-2.54(m,2H),2.67-2.72(m, 2H),3.58-3.64(m,1H),3.71-3.80(m,4H),5.03(s,2H),5.99-6.04(q,J=6.6Hz,1H),6.76(s,1H),7.04 -7.08(m,1H),7.28-7.32(m,1H),7.72(d,J=1.0Hz,1H).

Example 10: 3-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(3-(piperazin-1-yl)but-1-yne -1-yl)pyridin-2-amine

Step 1) N,N-bis(tert-butoxycarbonyl)-3-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy Base) -5-(3-(piperazin-1-yl)but-1-yn-1-yl)pyridin-2-amine

4-(6-(bis(tert-butoxycarbonyl)amino)-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl) But-3-yne-2-methanesulfonate (0.39 g, 0.6 mmol) was dissolved in acetonitrile (7 mL), and piperazine (0.52 g, 6 mmol) was added thereto. The reaction solution was stirred at 60°C for 16 hours, cooled to room temperature, and concentrated under reduced pressure. The residue was diluted with dichloromethane (50 mL). The resulting mixture was washed with water (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give the title compound as a yellow solid (0.23 g, 61%). The product was directly used in the next reaction without purification.

LC-MS (ESI, pos.ion) m/z: 637 [M+H] ⁺ .

Step 2) 3-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(3-(piperazin-1-yl)but-1-yne- 1- base) pyridin-2-amine

N,N-bis(tert-butoxycarbonyl)-3-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(3-(piperazine- 1-yl)but-1-yn-1-yl)pyridin-2-amine (0.2g, 0.3mmol) was dissolved in dichloromethane (5mL), and hydrogen chloride in ethyl acetate (3M, 1mL) was added ). After the reaction solution was stirred at room temperature for 18 hours, it was quenched with saturated potassium carbonate aqueous solution (30 mL), and extracted with ethyl acetate (50 mL x 3). The combined organic phases were washed successively with water (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DMC/MeOH/Et ₃ N (v/v/v)=200/20/1) to obtain the title compound as a yellow solid (123 mg, 90%).

LC-MS (ESI, pos.ion) m/z: 219[M/2+H] ⁺ ;

¹ H NMR(400MHz, CDCl ₃ )δ(ppm):1.37-1.39(m,3H),1.81(d,J=6.64Hz,3H),2.50-2.53(m,2H),2.67-2.71(m, 2H),2.91-2.98(m,4H),3.60-3.64(m,1H),4.96(s,2H),5.99-6.04(q,J=6.64Hz,1H),6.77(d,J=1.5Hz , 1H), 7.09-7.05(m, 1H), 7.32-7.26(m, 1H), 7.73(d, J=1.6Hz, 1H).

Example 11: 3-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(3-(4-methylpiperazin-1-yl) But-1-yn-1-yl)pyridin-2-amine

Step 1) N,N-bis(tert-butoxycarbonyl)-3-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(3-(4 - Methylpiperazin-1-yl)but-1-yn-1-yl)pyridin-2-amine

4-(6-(bis(tert-butoxycarbonyl)amino)-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl) But-3-yne-2-methanesulfonate (0.35 g, 0.55 mmol) was dissolved in acetonitrile (4 mL), and 1-methylpiperazine (0.22 g, 2.18 mmol) was added thereto. The reaction solution was stirred at 60°C for 16 hours, cooled to room temperature, and concentrated under reduced pressure. The residue was diluted with dichloromethane (50 mL), and the resulting mixture was washed with water (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE/EtAOc (v/v)=2/1) to obtain the title compound as a yellow solid (0.31 g, 94%).

LC-MS (ESI, pos. ion) m/z: 651 [M+H] ⁺ .

Step 2) 3-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(3-(4-methylpiperazin-1-yl)butan- 1- Alkyn-1-yl)pyridin-2-amine

N,N-bis(tert-butoxycarbonyl)-3-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(3-(4-methyl Piperazin-1-yl)but-1-yn-1-yl)pyridin-2-amine (0.31g, 0.47mmol) was dissolved in dichloromethane (5mL), and a solution of hydrogen chloride in ethyl acetate was added thereto (3M, 1 mL). After the reaction solution was stirred at room temperature for 18 hours, it was quenched with saturated potassium carbonate aqueous solution (30 mL), and extracted with ethyl acetate (50 mL x 3). The combined organic phases were washed successively with water (50 mL) and brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE/EtOAc (v/v)=1/1) to obtain the title compound as a yellow solid (182 mg, 91%).

LC-MS (ESI, pos.ion) m/z: 451[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ )δ(ppm): 1.38-1.40(dd, J=7.0Hz, 1.0Hz, 3H), 1.80(d, J=6.7Hz, 3H), 2.30(s, 3H), 2.50-2.58(m,6H),2.74-2.78(m,2H),3.62-3.67(q,J=7.0Hz,1H),4.91(s,2H),5.98-6.03(q,J=6.64Hz, 1H), 6.77(s, 1H), 7.08-7.04(m, 1H), 7.32-7.26(m, 1H), 7.72(d, J=1.6Hz, 1H).

Example 12: 5-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl) But-3-yn-2-yl)-5-azaspiro[2.4]heptane-7-amine

Step 1) (E)-5-Benzyl-7-(oximino)-5-azaspiro[2.4]heptan-4-one

5-Benzyl-5-azaspiro[2.4]heptane-4,7-dione (6 g, 28 mmol) and triethylamine (6 mL, 42 mmol) were dissolved in ethanol (144 mL), and hydrochloric acid was added thereto Hydroxylamine (2.92 g, 42 mmol). After stirring the reaction solution at room temperature for 24 hours, it was concentrated under reduced pressure. The residue was diluted with water (140 mL), and the resulting mixture was stirred at room temperature for 2 hours and then filtered. The filter cake was dried under reduced pressure at 45°C for 24 hours to obtain the title compound as a pale yellow solid (5.9 g, 91%).

LC-MS (ESI, pos.ion) m/z: 231.0[M+H] ⁺ ;

¹ H NMR(400MHz,d ₆ -DMSO)δ(ppm):1.22(d,J=3.6Hz,2H),1.35(d,J=3.6Hz,2H),4.02(s,2H),4.52(s ,2H),7.24-7.32(m,3H),7.35-7.39(m,2H).

Step 2) 5-Benzyl-5-azaspiro[2.4]heptane-7-amine

Lithium aluminum hydride (0.87g, 22.8mmol) was dissolved in tetrahydrofuran (10mL), cooled to 0°C, and (E)-5-benzyl-7-(hydroxyimino)-5-azaspiro was added thereto [2.4] A solution of heptan-4-one (1.31 g, 5.7 mmol) in tetrahydrofuran (20 mL). After the reaction solution was stirred at room temperature for 30 minutes, it was heated to 60° C. and stirred for 5 hours. After the reaction was completed, it was cooled to room temperature, and ethyl acetate (40 mL) and water (20 mL) were slowly added to quench the reaction. The mixture was filtered, the filtrate was concentrated under reduced pressure, the residue was diluted with ethyl acetate (40 mL), the resulting mixture was washed with water (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the title compound as an orange liquid (1.24 g, 99%).

LC-MS (ESI, pos.ion) m/z: 203.1[M+H] ⁺ ;

Step 3) 5-Benzyl-N-(tert-butoxycarbonyl)-5-azaspiro[2.4]heptane-7-amine

Dissolve 5-benzyl-5-azaspiro[2.4]heptane-7-amine (1.2g, 6.0mmol), triethylamine (2.1mL, 15mmol) in tetrahydrofuran (20mL), and add Boc Anhydride (2.1 mL, 9.0 mmol). After stirring the reaction solution at room temperature for 2 hours, it was concentrated under reduced pressure. The residue was diluted with dichloromethane (60 mL x 2), and the resulting mixture was washed with saturated brine (30 mL x 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether) to obtain the title compound as a white solid (1.85 g, 100%).

LC-MS (ESI, pos.ion) m/z: 303.1[M+H] ⁺ ;

¹ H NMR(400MHz,CDCl ₃ )δ(ppm):0.43-0.45(m,2H),0.58-0.80(m,2H),1.35(s,9H),2.34(d,J=8.8Hz,1H) ,2.66-2.69(m,2H),2.88-2.95(m,1H),3.60(dd,J=28.4Hz,12.8Hz,2H),3.88(s,1H),4.97(d,J=7.6Hz, 1H), 7.26(s, 2H), 7.30(s, 3H).

Step 4) N-(tert-butoxycarbonyl)-5-azaspiro[2.4]heptane-7-amine

5-Benzyl-N-(tert-butoxycarbonyl)-5-azaspiro[2.4]heptane-7-amine (0.93 g, 3 mmol) was dissolved in methanol (20 mL), and catalyst Pd/C ( 10%, 54.5% water), the reaction was fed with hydrogen, stirred at room temperature for 24 hours, filtered, and the filtrate was concentrated under reduced pressure to obtain the title compound as a yellow liquid (0.6 g, 100%).

LC-MS (ESI, pos.ion) m/z: 213.1[M+H] ⁺ ;

¹ H NMR(400MHz, CDCl ₃ )δ(ppm):0.56(dd,J=33.2Hz,9.6Hz,2H),0.77(s,2H),1.37(s,9H),2.73(d,J=10.8 Hz,1H),2.93(dd,J=11.6Hz,3.2Hz,1H),2.99(d,J=10.8Hz,1H),3.33(dd,J=11.6Hz,5.6Hz,1H),3.47(s ,1H), 3.66(s,1H), 7.26(s,2H), 4.75(s,1H).

Step 5) 5-(4-(6-(bis(tert-butoxycarbonyl)amino)-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy Base) pyridin-3-yl) but-3-yn-2-yl)-N-(tert-butoxycarbonyl)-5-azaspiro[2.4]heptane-7-amine

4-(6-(bis(tert-butoxycarbonyl)amino)-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl) But-3-yne-2-methanesulfonate (0.16g, 0.25mmol) was dissolved in acetonitrile (5mL), and N-(tert-butoxycarbonyl)-5-azaspiro[2.4]heptane was added at room temperature- 7-Amine (78.5 mg, 0.38 mmol) and triethylamine (0.35 mL, 2.5 mmol). The reaction solution was stirred at 69°C for 16 hours, cooled to room temperature, and concentrated under reduced pressure. The residue was diluted with ethyl acetate (50 mL), and the resulting mixture was washed with water (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE/EtOAc (v/v)=1/2) to obtain the title compound as a yellow solid (0.11 g, 58%).

LC-MS (ESI, pos.ion) m/z: 764[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ) δ (ppm): 0.50-0.70 (m, 2H), 0.73-0.92 (m, 2H), 1.30-1.41 (m, 3H), 1.41-1.46 (m, 27H), 1.79(d,J=6.6Hz,3H),2.54-2.64(m,1H),2.69-2.86(m,2H),3.01-3.16(m,1H),3.75-3.76(m,1H),3.84- 3.89(m,1H),4.91-4.95(m,1H),5.97-6.02(q,J=6.45Hz,1H),7.06-7.10(m,2H),7.30-7.32(m,1H),8.10( d, J=1.5Hz, 1H).

Step 6) 5-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)butan-3- Alkyn-2-yl)-5-azaspiro[2.4]heptane-7-amine

5-(4-(6-(bis(tert-butoxycarbonyl)amino)-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridine-3 -yl)but-3-yn-2-yl)-N-(tert-butoxycarbonyl)-5-azaspiro[2.4]heptane-7-amine (0.4g, 0.52mmol) was dissolved in dichloromethane ( 15 mL), the temperature was lowered to 0°C, and a solution of hydrogen chloride in ethyl acetate (3M, 5 mL) was added thereto. After stirring the reaction solution at room temperature for 16 hours, it was concentrated under reduced pressure. The residue was diluted with water (50 mL), adjusted to pH=10 by adding saturated aqueous sodium carbonate solution, and extracted with dichloromethane (80 mL x 3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM/MeOH/Et ₃ N (v/v/v)=200/20/1) to obtain the title compound as a yellow solid (294 mg, 90%).

LC-MS (ESI, pos.ion) m/z: 464[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ) δ (ppm): 0.41-0.49 (m, 1H), 0.59-0.67 (m, 2H), 0.75-0.78 (m, 1H), 1.40 (d, J=6.9Hz, 3H),1.81(d,J=6.7Hz,3H),2.56-2.64(m,2H),2.89-2.80(m,1H),3.07-3.14(m,1H),3.20-3.25(m,1H) ,3.70-3.73(m,1H),4.95(s,2H),6.00-6.02(q,J=6.5Hz,1H),6.75(s,1H),7.10(t,J=8.6Hz,1H), 7.28-7.32 (m, 1H), 7.73 (d, J=1.5Hz, 1H).

Example 13: 5-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl) But-3-yn-2-yl)-5-azaspiro[2.4]heptan-7-ol

Step 1) 5-(4-(6-(bis(tert-butoxycarbonyl)amino)-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy Base) pyridin-3-yl) but-3-yn-2-yl)-5-azaspiro[2.4]heptane-7-ol

4-(6-(bis(tert-butoxycarbonyl)amino)-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl) But-3-yne-2-methanesulfonate (0.99g, 1.53mmol) was dissolved in acetonitrile (13mL), and 5-azaspiro[2.4]heptan-7-ol (259mg, 1.95mmol) and triethylamine (2mmol). The reaction solution was stirred at 69°C for 16 hours, cooled to room temperature, and concentrated under reduced pressure. The residue was diluted with ethyl acetate (50 mL), and the resulting mixture was washed with water (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE/EtOAc (v/v)=1/1 to 1/2) to obtain the title compound as a yellow solid (0.74 g, 74%).

LC-MS (ESI, pos.ion) m/z: 664[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ) δ (ppm): 0.60-0.72 (m, 2H), 0.73-0.80 (m, 1H), 0.90-0.98 (m, 1H), 1.41-1.48 (m, 21H), 1.79(d,J=6.8Hz,3H),2.92-3.10(m,2H),3.72-3.88(m,2H),5.99(m,1H),7.08-7.105(m,3H),8.10(d, J=1.7Hz,1H).

Step 2) 5-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)butan-3- Alkyn-2-yl)-5-azaspiro[2.4]heptan-7-ol

5-(4-(6-(bis(tert-butoxycarbonyl)amino)-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridine-3 -yl)but-3-yn-2-yl)-5-azaspiro[2.4]heptan-7-ol (0.42g, 0.6mmol) was dissolved in dichloromethane (15mL), cooled to 0°C, A solution of hydrogen chloride in ethyl acetate (3M, 8 mL) was added thereto. The reaction solution was stirred at room temperature for 16 hours, then concentrated under reduced pressure. The residue was diluted with water (50 mL), adjusted to pH=10 by adding saturated aqueous sodium carbonate solution, and extracted with dichloromethane (50 mL x 3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EtOAc/MeOH/Et ₃ N (v/v/v)=250/10/1) to obtain the title compound as a yellow solid (0.16 g, 60%).

LC-MS (ESI, pos.ion) m/z: 464[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ) δ (ppm): 0.57-0.58 (m, 1H), 0.63-0.65 (m, 1H), 0.72-0.75 (m, 1H), 0.92-0.94 (m, 1H), 1.39-1.42(m,3H),1.81(d,J=6.7Hz,3H),2.57-2.64(m,1H),2.84-3.00(m,3H),3.71-3.77(m,2H),5.01( s, 2H), 6.01(m, 1H), 6.75(d, J=1.4Hz, 1H), 7.05-7.09(m, 1H), 7.29-7.31(m, 1H), 7.72(s, 1H).

Example 14: 5-(3-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl) prop-2-yn-1-yl)-5-azaspiro[2.4]heptan-7-ol

Step 1) (R)-3-(6-(bis(tert-butoxycarbonyl)amino)-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy) Pyridin-3-yl)prop-2-yn-1-ol

(R)-5-bromo-N, N-bis(tert-butoxycarbonyl)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-2-amine ( 2g, 3.5mmol), bis(triphenylphosphine)palladium dichloride (246mg, 0.35mmol) and cuprous iodide (67mg, 0.35mmol) were suspended in triethylamine (18mL), and 2 - propyn-1-ol (0.42 mL, 7 mmol). The reaction solution was microwaved at 90° C. for 80 minutes, cooled to room temperature, and concentrated under reduced pressure. The residue was diluted with dichloromethane (100 mL), filtered, and the filtrate was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE/EtOAc (v/v)=2/1) to obtain the title compound as a yellow solid (1.52 g, 80%).

¹ H NMR (400MHz, CDCl ₃ ) δ (ppm): 1.37-1.41 (m, 18H), 1.55 (d, J=6.6Hz, 3H), 1.78 (d, J=6.6Hz, 3H), 4.72-4.78 (m,1H),5.97-5.02(q,J=6.7Hz,1H),7.06-7.12(m,2H),7.27-7.32(m,1H),8.12-8.13(m,1H).

Step 2) (R)-3-(6-(bis(tert-butoxycarbonyl)amino)-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy) Pyridin-3-yl)prop-2-yn-1-methanesulfonate

Under nitrogen protection, (R)-3-(6-(bis(tert-butoxycarbonyl)amino)-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridine- 3-yl)prop-2-yn-1-ol (300mg, 0.54mmol) was dissolved in dichloromethane (2.5mL), cooled to 0°C, and triethylamine (0.22mL, 1.6mmol) and formaldehyde were added thereto Sulfonyl chloride (63 μL, 0.81 mmol). After the reaction solution was stirred at room temperature for 1 hour, it was quenched with saturated aqueous sodium bicarbonate (10 mL), and extracted with dichloromethane (10 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure to afford the title compound. The product was directly used in the next reaction without purification.

LC-MS (ESI, pos.ion) m/z: 633 [M+H] ⁺ .

Step 3) 5-(3-(6-(bis(tert-butoxycarbonyl)amino)-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy Base) pyridin-3-yl) prop-2-yn-1-yl)-5-azaspiro[2.4]heptane-7-ol

(R)-3-(6-(bis(tert-butoxycarbonyl)amino)-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl) Prop-2-yne-1-methanesulfonate (341mg, 0.54mmol) was dissolved in acetonitrile (5mL), and 5-azaspiro[2.4]heptan-7-ol (92mg, 1.08mmol) was added thereto at room temperature ) and triethylamine (1 mL). The reaction solution was stirred at 69°C for 16 hours, cooled to room temperature, and concentrated under reduced pressure. The residue was diluted with ethyl acetate (50 mL), and the resulting mixture was washed with water (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (PE/EtOAc (v/v)=2/1) to give the title compound as a yellow solid (0.14 g, 40%).

LC-MS (ESI, pos.ion) m/z: 650[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ) δ (ppm): 0.61-0.69 (m, 2H), 0.77-0.78 (m, 1H), 0.93-0.96 (m, 1H), 1.38-1.43 (m, 18H), 1.78(d,J=6.6Hz,3H),2.57-2.60(dd,J=8.8Hz,4.8Hz,1H),2.92-2.95(m,2H),3.04-3.07(dd,J=10.0Hz,1.6 Hz,1H),3.65(s,2H),3.77-3.79(m,1H),5.99(m,1H),7.05-7.10(m,2H),7.27-7.32(m,1H),8.10(d, J=1.7Hz,1H).

Step 4) 5-(3-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)propan-2- Alkyn-1-yl)-5-azaspiro[2.4]heptan-7-ol

5-(3-(6-(bis(tert-butoxycarbonyl)amino)-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridine-3 -yl)prop-2-yn-1-yl)-5-azaspiro[2.4]heptan-7-ol (0.14g, 0.22mmol) was dissolved in dichloromethane (6mL), cooled to 0°C, A solution of hydrogen chloride in ethyl acetate (3M, 2 mL) was added thereto. After the reaction solution was stirred at room temperature for 16 hours, it was concentrated under reduced pressure. The residue was diluted with water (50 mL), adjusted to pH=10 by adding saturated aqueous sodium carbonate solution, and extracted with dichloromethane (50 mL x 3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EtOAc/MeOH/Et ₃ N (v/v/v)=300/30/1) to obtain the title compound as a yellow solid (85 mg, 88%).

LC-MS (ESI, pos.ion) m/z: 450[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ) δ (ppm): 0.69-0.78 (m, 3H), 0.91-0.95 (m, 1H), 1.81 (d, J=6.7Hz, 3H), 2.58 (d, J= 8.6Hz,1H),2.92(t,J=8.6Hz,2H),3.02-3.05(dd,J=10.1Hz,4.9Hz,1H),3.61(s,2H),3.75-3.76(m,1H) ,4.96(s,2H),5.98-6.03(q,J=6.7Hz,1H),6.74(d,J=1.3Hz,1H),7.05-7.09(m,1H),7.29-7.32(m,1H ), 7.73(s,1H).

biological test

The compounds prepared in the examples of the present invention were subjected to biological analysis using the methods and equipment described above.

Stability of Example A in Human and Rat Liver Microsomes

Human or rat liver microsomes are incubated in polypropylene tubes and primed for replication. A typical incubation mixture consists of human or rat liver microsomes (0.5 mg protein/mL), target compound (5 μM), and NADPH (1.0 mM) potassium phosphate buffer (PBS, 100 mM, pH 7.4) in a total volume of 200 μL ), the compounds were dissolved in DMSO and diluted with PBS so that the final concentration of the DMSO solution was 0.05%. And incubate at 37° C. in a water bath connected to air. After pre-incubation for 3 minutes, add protein to the mixture and start the reaction. At different time points (0, 5, 10, 15, 30 and 60 min), the same volume of ice-cold acetonitrile was added to terminate the reaction. Samples were stored at -80°C until LC/MS/MS analysis.

Compound concentrations in human or rat liver microsome incubation mixtures were determined by LC/MS/MS. The linear range of the concentration range was determined for each compound tested.

Parallel incubation experiments using denatured microsomes as negative controls were incubated at 37°C and the reactions were terminated at different time points (0, 15 and 60 min).

Dextromethorphan (70 μM) was used as a positive control, incubated at 37°C, and the reactions were terminated at different time points (0, 5, 10, 15, 30 and 60 minutes). Positive and negative control samples are included with each assay to ensure the integrity of the microsomal incubation system.

For each reaction, the in vivo liver intrinsic clearance CL _int (ref.:Naritomi Y, Terashita S, Kimura S, Suzuki A, Kagayama A, Sugiyama Y. Prediction of human hepatic clearance from in vivo animal experiments and in vitro metabolic studies with liver microsomes from animals and humans. Drug Metabolism and Disposition 2001, 29: 1316-1324.) After testing, the results showed that when the compounds provided in the examples of the present invention were respectively incubated in human and rat liver microsomes, the compounds of the present invention showed good half-life (T _1/2 ) and CL _int .

The stability test of the compound provided by the embodiments of the present invention in people and rat liver microsomes in table 1

test results

Embodiment B The pharmacokinetic evaluation of compound of the present invention in animal body

The present invention evaluates the pharmacokinetic studies of the compounds of the present invention in mice, rats, dogs or monkeys.

The compound of the present invention is administered in the form of aqueous solution, 2% HPMC+1% Tween-80 aqueous solution, 5% DMSO+5% saline solution. Blood (0.3 mL) was drawn at time points 0.25, 0.5, 1.0, 2.0, 3.0, 4.0, 6.0, 8.0, 12 and 24 hours and centrifuged at 3,000 or 4,000 rpm for 10 minutes. Plasma solutions were collected and stored at -20°C or -70°C until LC/MS/MS analysis as described above. See Table 2 for the results. Table 2 is the experimental results of the pharmacokinetic characteristics of the compounds provided in the examples of the present invention in rats.

Table 2 The pharmacokinetic characteristics experimental results of the compounds provided by the embodiments of the present invention in rats

The results show that when the compound is administered intravenously, the compound of the present invention exhibits good pharmacokinetic properties, including ideal clearance rate (Cl), half-life (T _1/2 ) and good oral bioavailability.

Example C Kinase Assay

According to the method described above, the ALK(h) kinase assay and c-Met(h) kinase assay were carried out on the compounds provided in the examples of the present invention, and the results are shown in Table 3, which is the results of the kinase assays provided in the examples of the present invention.

The kinase assay result of the compound that the embodiment of the present invention provides in table 3

It can be known from Table 3 that the compounds of the present invention generally show very high activity in the tests of ALK and c-Met(h).

Example D Cell Phosphorylation

According to the method described above, the cellular phosphorylation levels of the compounds provided in the examples of the present invention were detected, and the IC ₅₀ values were estimated (each concentration was measured twice). The test results show that the compound of the present invention generally shows very high activity in the cell phosphorylation test of ALK and c-Met.

Example E Xenograft Tumor Model

The U87MG transplanted tumor model was established by the method described above, and analyzed by the method described above. In the U87MG xenograft tumor model, the compounds of Examples 1 to 14 were orally administered (p.o.) daily (QD) for 13-21 days. At a dose of 60 mg/kg, the compounds of Examples 1 to 14 all have statistical significance and can inhibit the growth of subcutaneous tumors in nude mice.

Finally, it should be noted that there are other ways to implement the invention. Accordingly, the embodiments of the present invention will be described as illustrations, but are not limited to the content described in the present invention, and may be modified within the scope of the present invention or equivalent content added in the claims. All publications or patents cited in the present invention shall be regarded as reference documents of the present invention.

Claims (15)

1. a compound as shown in the formula (I):

Or its steric isomer, geometrical isomer, tautomer, oxynitride, solvate, meta-bolites, pharmacy acceptable salt or its prodrug, wherein:

Each R ¹, R ², R ³, R ⁴, R ⁵and R ⁶be H independently, D or F;

X is C _6-10aryl or comprise 1,2,3 or 4 and independently be selected from O, the heteroaryl of heteroatomic 5-10 the atom of S or N, wherein, described C _6-10aryl and comprise 1,2,3 or 4 and independently be selected from O, the heteroaryl of heteroatomic 5-10 the atom of S or N is optionally by 1,2, and 3 or 4 substituting groups replace, and described substituting group is independently selected from D, F, Cl, Br, I ,-CN ,-NO ₂, N ₃,-OR ^a,-SR ^a,-NR ^ar ^b,-C (=O) NR ^ar ^b, C _1-6alkyl, C _1-6haloalkyl, C _2-6alkenyl, C _2-6alkynyl ,-(C _1-4alkylidene group)-CN ,-(C _1-4alkylidene group)-OR ^a,-(C _1-4alkylidene group)-NR ^ar ^b, C _6-10aryl or comprise 1,2,3 or 4 and independently be selected from O, the heteroaryl of heteroatomic 5-10 the atom of S or N;

Z is C _1-6alkyl, C _3-6cycloalkyl, C _3-6heterocyclic radical, C _5-12the condensed-bicyclic base, C _5-12the spiral shell bicyclic group ,-(C _1-4alkylidene group)-(C _3-6cycloalkyl) ,-(C _1-4alkylidene group)-(C _3-6heterocyclic radical) ,-(C _1-4alkylidene group)-(C _5-12the condensed-bicyclic base) or-(C _1-4alkylidene group)-(C _5-12the spiral shell bicyclic group), wherein, described C _1-6alkyl, C _3-6cycloalkyl, C _3-6heterocyclic radical, C _5-12the condensed-bicyclic base, C _5-12the spiral shell bicyclic group ,-(C _1-4alkylidene group)-(C _3-6cycloalkyl) ,-(C _1-4alkylidene group)-(C _3-6heterocyclic radical) ,-(C _1-4alkylidene group)-(C _5-12the condensed-bicyclic base) and-(C _1-4alkylidene group)-(C _5-12the spiral shell bicyclic group), optionally by 1,2,3 or 4 substituting groups replace, and described substituting group is independently selected from D, F, Cl, Br, I ,-CN ,-NO ₂, N ₃, C _1-4alkyl ,-OR ^a,-SR ^a,-NR ^ar ^bor-C (=O) NR ^ar ^b, and, C _5-12the condensed-bicyclic base, C _5-12the spiral shell bicyclic group ,-(C _1-4alkylidene group)-(C _5-12the condensed-bicyclic base) or-(C _1-4alkylidene group)-(C _5-12the spiral shell bicyclic group) each ring in is carbocyclic ring or heterocycle independently;

Each R ^aand R ^bbe H independently, C _1-6aliphatics, C _3-6cycloalkyl, C _3-6heterocyclic radical, C _6-10aryl, comprise 1,2,3 or 4 and independently be selected from O, the heteroaryl of heteroatomic 5-10 the atom of S or N ,-(C _1-4alkylidene group)-(C _3-6cycloalkyl) ,-(C _1-4alkylidene group)-(C _3-6heterocyclic radical) ,-(C _1-4alkylidene group)-(C _6-10aryl) or-(C _1-4alkylidene group)-(comprise 1,2,3 or 4 and independently be selected from O, the heteroaryl of heteroatomic 5-10 the atom of S or N), and, R worked as ^aand R ^bwhile being connected with same nitrogen-atoms, R ^a, R ^b, together with the nitrogen-atoms be connected with them, can also optionally form the heterocyclic radical of 3-8 atom, wherein, described C _1-6aliphatics, C _3-6cycloalkyl, C _3-6heterocyclic radical, C _6-10aryl, comprise 1,2,3 or 4 and independently be selected from O, the heteroaryl of heteroatomic 5-10 the atom of S or N ,-(C _1-4alkylidene group)-(C _3-6cycloalkyl) ,-(C _1-4alkylidene group)-(C _3-6heterocyclic radical) ,-(C _1-4alkylidene group)-(C _6-10aryl) ,-(C _1-4the heterocyclic radical of alkylidene group)-(comprise 1,2,3 or 4 and independently be selected from O, the heteroaryl of heteroatomic 5-10 the atom of S or N) and 3-8 atom is optionally by 1,2, and 3 or 4 substituting groups replace, and described substituting group is independently selected from D, F, Cl ,-CN, N ₃,-OH ,-NH ₂, alkoxyl group or alkylamino;

Situation distinguishingly, when Z is C _1-6during alkyl ,-NR ^ar ^bcan not be-NH ₂or-NMe ₂.

2. compound according to claim 1, wherein, each R ¹, R ², R ³, R ⁴, R ⁵and R ⁶be H or D independently.

3. compound according to claim 1, wherein, X is phenyl, and can be optionally by 1,2,3 or 4 independently are selected from D, F, Cl, Br, C _1-3alkyl or C _1-3the substituting group of haloalkyl replaces.

4. compound according to claim 1, wherein, Z is C _1-6alkyl, C _3-6cycloalkyl, C _3-6heterocyclic radical, C _5-12the condensed-bicyclic base, C _5-12the spiral shell bicyclic group ,-(C _1-4alkylidene group)-(C _3-6heterocyclic radical) ,-(C _1-4alkylidene group)-(C _5-12the condensed-bicyclic base) or-(C _1-4alkylidene group)-(C _5-12the spiral shell bicyclic group), wherein, described C _1-6alkyl, C _3-6cycloalkyl, C _3-6heterocyclic radical, C _5-12the condensed-bicyclic base, C _5-12the spiral shell bicyclic group ,-(C _1-4alkylidene group)-(C _3-6cycloalkyl) ,-(C _1-4alkylidene group)-(C _3-6heterocyclic radical) ,-(C _1-4alkylidene group)-(C _5-12the condensed-bicyclic base) and-(C _1-4alkylidene group)-(C _5-12the spiral shell bicyclic group), optionally by 1,2,3 or 4 independently selected from D, F, Cl, Br, I ,-CN ,-NO ₂, N ₃, C _1-3alkyl ,-OR ^a,-NR ^ar ^bsubstituting group replace, and, C _5-12the condensed-bicyclic base, C _5-12the spiral shell bicyclic group ,-(C _1-4alkylidene group)-(C _5-12the condensed-bicyclic base) or-(C _1-4alkylidene group)-(C _5-12the spiral shell bicyclic group) each ring in is carbocyclic ring or heterocycle independently.

5. compound according to claim 1, wherein, each R ^aand R ^bbe H independently, C _1-3alkyl, C _3-6cycloalkyl or-(C _1-4alkylidene group)-(C _3-6and work as R cycloalkyl), ^aand R ^bwhile being connected with same nitrogen-atoms, R ^a, R ^b, together with the nitrogen-atoms be connected with them, can also optionally form the heterocyclic radical of 3-6 atom, wherein, described C _1-3alkyl, C _3-6cycloalkyl or-(C _1-4alkylidene group)-(C _3-6cycloalkyl) and the heterocyclic radical of 3-6 atom optionally by 1,2,3 or 4 substituting groups replace, described substituting group is independently selected from D, F, Cl ,-OH ,-NH ₂, C _1-3alkoxyl group or C _1-3alkylamino;

Special situation, when Z is C _1-6during alkyl ,-NR ^ar ^bcan not be-NH ₂or-NMe ₂.

6. compound according to claim 1, wherein, Z is selected from any one in minor structure shown in formula (Z1)～(Z42):

Or its steric isomer, wherein:

N is 0,1,2 or 3;

Each W and W ' be independently-O-or-N (H)-;

Minor structure shown in formula (Z1)～(Z42) or its steric isomer be optionally by 1,2, and 3 or 4 independently selected from D, F, Cl, Br, I ,-CN ,-NO ₂, N ₃, C _1-3alkyl ,-OR ^a,-NR ^ar ^bsubstituting group replace, wherein, when Z is the minor structure shown in formula (Z25)～(Z32) or its steric isomer ,-NR ^ar ^bcan not be-NH ₂or-NMe ₂.

7. compound according to claim 1 has following one of them structure:

8. a pharmaceutical composition, comprise the described compound of claim 1-7 any one and pharmaceutically acceptable carrier, vehicle, thinner, assistant agent, vehicle or their combination.

9. pharmaceutical composition according to claim 8, also comprise the additional treatment agent, and described additional treatment agent is selected from chemotherapeutic agent, and antiproliferative is used for the treatment of atherosclerotic medicine, is used for the treatment of the medicine of pulmonary fibrosis or their combination.

10. pharmaceutical composition according to claim 9, described additional treatment agent is Chlorambucil (chlorambucil), melphalan (melphalan), endoxan (cyclophosphamide), ifosfamide (ifosfamide), busulfan (busulfan), carmustine (carmustine), lomustine (lomustine), streptozotocin (streptozocin), cis-platinum (cisplatin), carboplatin (carboplatin), oxaliplatin (oxaliplatin), Dacarbazine (dacarbazine), Temozolomide (temozolomide), Procarbazine (procarbazine), methotrexate (methotrexate), Fluracil (fluorouracil), cytosine arabinoside (cytarabine), gemcitabine (gemcitabine), purinethol (mercaptopurine), fludarabine (fludarabine), vinealeucoblastine(VLB) (vinblastine), vincristine(VCR) (vincristine), vinorelbine (vinorelbine), taxol (paclitaxel), Docetaxel (docetaxel), topotecan (topotecan), irinotecan (irinotecan), Etoposide (etoposide), ET-743 (trabectedin), gengshengmeisu (dactinomycin), Dx (doxorubicin), epirubicin (epirubicin), daunomycin (daunorubicin), mitoxantrone (mitoxantrone), bleomycin (bleomycin), ametycin (mitomycin), ipsapirone (ixabepilone), tamoxifen (tamoxifen), flutamide (flutamide), gonadorelin analogue (gonadorelin analogues), megestrol (megestrol), prednisone (prednidone), dexamethasone (dexamethasone), methylprednisolone (methylprednisolone), Thalidomide (thalidomide), interferon alpha (interferon alfa), Calciumlevofolinate (leucovorin), sirolimus (sirolimus), temsirolimus (temsirolimus), everolimus (everolimus), Ah method is for Buddhist nun (afatinib), alisertib, amuvatinib, A Pa is for Buddhist nun (apatinib), Axitinib (axitinib), Velcade (bortezomib), SKI-606 (bosutinib), brivanib, cabozantinib, AZD2171 (cediranib), crenolanib, Ke Zhuo is for Buddhist nun (crizotinib), dabrafenib, dacomitinib, danusertib, Dasatinib (dasatinib), dovitinib, Tarceva (erlotinib), foretinib, ganetespib, Gefitinib (gefitinib), ibrutinib, Conmana (icotinib), imatinib (imatinib), iniparib, lapatinibditosylate (lapatinib), lenvatinib, linifanib, linsitinib, Masitinib (masitinib), momelotinib, not for husky Buddhist nun (motesanib), HKI-272 (neratinib), nilotinib (nilotinib), niraparib, oprozomib, olaparib, pazopanib (pazopanib), pictilisib, ponatinib, quizartinib, regorafenib, rigosertib, rucaparib, ruxolitinib, fork clip is for Buddhist nun (saracatinib), saridegib, Xarelto (sorafenib), Sutent (sunitinib), tasocitinib, telatinib, tivantinib, tivozanib, tofacitinib, trametinib, ZD6474 (vandetanib), veliparib, Wei Luofeini (vemurafenib), vismodegib, volasertib, alemtuzumab (alemtuzumab), rhuMAb-VEGF (bevacizumab), brentuximab vedotin, block appropriate rope monoclonal antibody (catumaxomab), Cetuximab (cetuximab), ground promise monoclonal antibody (denosumab), lucky trastuzumab (gemtuzumab), her monoclonal antibody (ipilimumab), Buddhist nun's trastuzumab (nimotuzumab), method wood monoclonal antibody (ofatumumab) difficult to understand, Victibix (panitumumab), Rituximab (rituximab), tositumomab (tositumomab), Herceptin (trastuzumab), or their combination.

11. a right to use requires the described compound of 1-7 any one or the described pharmaceutical composition of claim 8-10 any one for the preparation of the purposes of protecting, process, treat or alleviate the medicine of patient's proliferative disease.

12. the purposes according to the described compound of claim 11 or pharmaceutical composition, is characterized in that, described proliferative disease is metastatic carcinoma, colorectal carcinoma, adenocarcinoma of stomach, bladder cancer, mammary cancer, kidney, liver cancer, lung cancer, skin carcinoma, thyroid carcinoma, brain tumor, neck cancer, prostate cancer, carcinoma of the pancreas, the cancer of central nervous system, glioblastoma, myeloproliferative disease, atherosclerosis or pulmonary fibrosis.

13. a right to use requires the described compound of 1-7 any one or the described pharmaceutical composition of claim 8-10 any one to come for the preparation of suppress or regulate the purposes of the medicine of protein kinase activity in biological sample, described purposes comprises right to use and requires the described compound of 1-7 any one or right to use to require the described pharmaceutical composition of 8-10 any one to contact with described biological sample.

14. purposes according to claim 13, is characterized in that, described protein kinase is receptor tyrosine kinase.

15. purposes according to claim 14, is characterized in that, described receptor tyrosine kinase is ALK, c-Met, or their combination.